PMID- 18411624 TI - [Technical aspects of isolated organ perfusion devices]. AB - An aspect of capital importance in the transplant field is the maintainance and preservation of organs during the ischemic phase. Organ susceptibility and injuries secondary to ischemia, organ procurement and transportation are still nowadays one of the main causes for organ failure. Effective artificial organ perfusion has remained a main goal for researchers in this field for more than 100 years. In the second half of the XXth century these techniques have been applied to experimental and clinical organ preservation, with the aim of extending the period of an effective storage (which guarantees an early and good function after the transplant). Primary goal of any organ preservation is to maintain integrity in every cell system in order to minimize those injuries which produces graft dysfunction. PMID- 18411625 TI - [Pharmacological preconditioning with sildenafil of warm ischemic kidneys]. AB - OBJECTIVES: To evaluate the preconditioning effect of sildenafil administered preoperatively in kidneys subjected to a period of warm ischemia (WI), hypothermic perfusion (HP) or cold storage (CS) and finally, autotransplant (AT). MATERIAL AND METHOD: We studied 6 groups of autotransplanted kidneys: no-WI inmediate AT (Group A); 45 min of WI + immediate AT (Group B); 45 min of WI + 60 min of HP + autotransplant (Group C); 45 min of WI + 60 min of CS + autotransplant (Group D); 100 mg of oral sildenafil preoperatively + 45 min of WI + autotransplant (Group E); 100 mg of oral sildenafil preoperatively + 45 min of WI+60 min of HP + autotransplant (Group F). Belzer solution was used for HP; UW Viaspan for CS. Inmediately after the autotransplant (reperfusion period), we recorded in real time for 60 min the values of Renal vascular Flow (RVF) and Renal Vascular Resistance (RVR). Nitric Oxide levels in the cava and renal graft vein were recorded every 15 min during the 60 min of the reperfusion-study period. Conventional & Electronic microscopy were completed after the process. RESULTS: We obtained significant higher values of RVF and lower values of RVR in sildenafil groups (E and F) in comparison to the other groups (A-D) (Table 1). NO levels were also significantly higher in groups E and F (Fig. 1). Groups A, B, E and F showed integrity of tubule and endothelium in comparison to groups C and D in the microscopic study. CONCLUSIONS: We showed a beneficious effect of sildenafil in inmediate post-transplant reperfusion hemodynamic and biochemical parameters of kidneys subjected to a critical period of warm-ischemia. PMID- 18411626 TI - [Hystologic and hemodynamic aspects of warm ischemic graft in relation to the preservation method]. AB - OBJECTIVE: The non-heart-beating donor has been proposed as a solution to donor shortage for renal transplantation. Because the nature of such donors, the kidneys so derived have been damaged by primary warm ischemia (WI), and so potentially they may never function. Minimizing graft injury is especially important in case of transplantation form marginal donors because of a high rate of delayed graft function or primary nonfunction. The aim of this experimental study is to assess the structural and hemodynamic consequences of hypothermic perfusion (HP) versus cold storage (CS), in renal allograft after a period of WI. MATERIAL AND METHODS: We used 20 mini-pigs. WI was achieved by vascular pedicle occlusion during 45 min. We divided organs in 4 groups: A (n=5), kidneys with WI and then transplanted; group B (n=5), grafts with WI and implanted after HP with Belzer solution in our computerized perfusion system. Group C-control, (n=5) transplanted without WI and D (n=5) with WI and 60 min of CS in UW-Viaspan solution. All the procedure was recorded by a computerized data system. Renal vascular resistance (RVR) and renal vascular flow (RVF) were automatically calculated by means of mathematical formulas after renal transplantation. Subsequently histological study was completed in all cases. RESULTS: We observed two patterns after transplantation: (1). Initial increase of RVR with posterior decrease and increase of vascular flow: in organs with WI and HP prior to transplantation (group B) // organs transplanted without WI (group C-control). Electronic and conventional microscopy showed integrity of endothelial and tubule structure. (2). Initial decrease with posterior increase of RVR. Organs with WI (group A) // organs with WI and CS (group D). Structural study showed endothelial and tubule disruption. CONCLUSION: In our experimental model machine perfusion preserves endothelial and tubule structure of kidneys with WI. After transplantation the hemodynamic pattern of grafts with WI and HP is similar to the control group (without WI and direct transplantation). PMID- 18411627 TI - [Experimental models for research and training in renal transplant]. AB - An update on aspects and use of different experimental models applied in kidney transplant research is presented . This paper includes qualities, as long as similarities between most frequently used animal models and human clinical standards. Contributions of those models based on microsurgical or laparoscopic techniques are revised. The physiological consequences (hemodynamic, immunologic) of surgical technique (laparoscopy), applied in experimental models as long as non-heart beating organ donor models and organ preservation methods are also reviewed. Finally, an update of those models applied in research in prothocols of either immunosupression or xenotransplant is done. PMID- 18411628 TI - [Pig model in experimental renal transplant surgery]. AB - INTRODUCTION AND OBJECTIVES: Living donor laparoscopic renal procurement is becoming a first-line technique unless a show-learning curve. January 2006 we implement an experimental pig-kidney transplant model with the objective of evaluating differences between open and laparosopic surgical techniques as well as giving a training-oportunity to the Residents in these alternatives. MATERIAL AND METHODS: We have completed 25 experiments 7 out of which were performed laparoscopically (28%), 18 with conventional surgerY (72%). Only 44% of the animals have survived until the end of the process. RESULTS: This work evaluates different aspects on the implementation of this activity. Complications of the prothocol are analyzed. We review the literature on this topic. CONCLUSIONS: Experimental Surgery in a porcine model has become in our Hospital a key-issue for Residents Training Program, and easily could be adapted to other Centers. PMID- 18411629 TI - [Experimental pancreatic islet transplant into the genito-urinary tract simultaneous to kidney transplant]. AB - INTRODUCTION AND OBJECTIVES: Simultaneous kidney and pancreas transplant is a good treatment for both renal and pancreas insufficiency. Experimental apply of genitourinary tract for pancreas implantation is reported in this work. MATERIAL AND METHOD: Twenty animals aged as average 5.5 monts (SD 1.1) and an average weight of 53 kgr were submitted to this protocol. In the day 1 a left nephrectomy is completed and the graft is perfused with University of Wisconsin solution. A partial pancreatectomy is completed at following, isolation of pancreatic islets by colagenase enzymatic digestion. Islets are dryed with Ditizone and culptured for 24 hours at 37 degrees C and 5% CO2. Day-2 a right nephrectomy is performed and orthotopic renal autotransplant using the left kidney is completed. Pancreatic islets are transplanted in 4 different locations of the genitourinary tract: renal subcapsular space, bladder submucosae, testis parenchyma and vas deferens. Day-7, all the animals were sacrifized to complete pathological study. RESULTS AND CONCLUSIONS: Viable islets were isolated in bladder submucosae and testis after transdeferential injection. PMID- 18411630 TI - [Ischemia-reperfusion syndrome and role of preservation graft technique after laparoscopic versus open nephrectomy in a experimental model of living donor kidney transplant]. AB - INTRODUCTION: Delayed graft function alter living donor transplantation is a subject of debate. Delayed graft function can be partially explained by renal ischemia-reperfusion injury, when severe is associated with decreased graft survival. In this experimental living donor model study, we analyze the hemodynamic, histological and biochemical effects of laparoscopic nephrectomy. We also, analyze the effect of a pulsatile machine perfusion for kidney preservation during cold ischemia time. MATERIAL AND METHODS: Twenty large-white pigs (average weight 40-45 kgrs) were divided in 4 experimental groups: Group A: Laparoscopic nephrectomy+ immediate graft perfusion in pulsatile vacuum pump+autotransplant Group B: Laparoscopic nephrectomy+ immediate graft perfusion by gravity+autotransplant Group C: Open nephrectomy+immediate graft perfusion in pulsatile vacuum pump+autotransplant Group D: Open nephrectomy+ immediate graft perfusion by gravity+autotransplant Both laparoscopic and open nephrectomy were completed transperitoneally according to standardized technique. Hypothermic perfusion was done in a system designed in our lab. RESULTS: We observed a decreased renal artery flow in kidneys procured laparoscopically compared to open nephrectomy. We found an artery flow recovery during the first 60 minutes after revascularization. Renal machine perfusion during cold ischemia time seems to have no beneficial effect, but shows a deleterious effect on hemodynamic event for renal transplantation. Lower plasma nitric oxide level is observed in kidneys obtained by laparoscopy compared with open surgical technique. And finally, we also found higher histological damage in proximal tubular and endothelial cell, in kidneys obtained by laparoscopy compared with open surgery. CONCLUSIONS: In our experience: Laparoscopic nephrectomy versus open nephrectomy produces, in a model of living donor transplant, a lower value or renal blood flow and a higher value of renal vascular resistanse. These hemodynamic findings tend to normalize by 60 min after the reperfusion. A lower blood concentration of nitric oxide after the transplant was detected in laparoscopic group Vs open surgery group. PMID- 18411631 TI - [Study on the role of the tubule in renal vasoconstriction induced by cyclosporine]. AB - INTRODUCTION: Cyclosporine (CyA) has proved to induce cell apoptosis on cultured proximal tubule cells. However, there is no much data about the in vivo functional consequences of this injury or the long time observed CyA-induced renal vasoconstriction. MATERIAL AND METHODS: In a swine model of subacute CyA nephrotoxicity (10 mg/ Kg. dx 15 days), we performed a right nephrectomy, followed by left renal artery, vein and ureter catheterisati8n. After inducing water diuresis, three clearance periods of 15 minutes were performed before and after a furosemide 1 mg/kg infusion. Plasma and urine electrolytes, blood gas, acid excretion, plasma renin activity and aldosterone concentration, GFR, RPF, RBF, intra-renal vascular resistances, glomerular filtration pressure, distal Cl- delivery, water clearance and TTKG were measured or estimated on 7 control and 7 treated animals. Right kidney was processed for NaKATPase activity and immunostaining. RESULTS: Treated animals presented detaching proximal cells, luminal blebbing and loss of tight junctions. Cortical but not medullar sodium pump was internalised and partially inactive. Treated animals showed much lower fractional excretions of Na+, with significantly higher distal fractional reabsorption of Cl. Distal shift in fluid load resulted in a significant rise in renal O2 consumption, and modifications in the global renal estequiometry of Na+ transport/O2 uptake. Several consequences followed this situation: preglomerular resistances increased 3 times with only minor changes in postglomerular resistances and renal blood and plasma flow were significantly reduced. Furosemide partially reversed these effects. A slight increase in fractional filtration prevented GFR differences to become statistically significant. CONCLUSION: subacute CyA treatment even al doses not modifying GFR, may cause proximal tubule Na+ transport impairment, resulting in increased rates of distal delivery and absorption of fluid load. Renal uptake of O2 may be increased and tubule glomerular feedback should be expected to be activated. Absence of changes of GFR with furosemide is an early sign of CyA renal damage. PMID- 18411632 TI - [Comparative analysis of renal graft function after open vs. laparoscopic nephrectomy: experimental model]. AB - INTRODUCTION: Living donor renal transplant reports a higher patient and graft survival in comparison to cadaver donor and represents a good alternative facing the current lack of organs for transplant. GOALS: To analyze comparatively in an experimental model (pig) the influence of ischemia-reperfusion and functional outcome of renal graft retrieved by open Vs laparoscopic nephrectomy. MATERIAL AND METHODS: 30 lab pigs were nephrectomized (left kidney): 15 by laparoscopy and 15 by open surgery, as living donors, in a model of renal autotransplant. Renal blood flow (RBF) was measured by means of an electromagnetic probe and creatinine levels during the first week after the implant. RESULTS: Comparative analysis of RBF during the immediate 60 min after unclamping showed a significant reduction of average RBF in laparoscopic group in comparison to open group (p < 0.001), with a more evident reduction of RBF in the laparoscopic group during the 5-min period after unclamping (p < 0.001) and a progressive recuperation of RBF during the 1st hour, slowest in laparoscopic group. Creatinine levels in the first week after the transplant decreased progressively from 1.3 to 0.8 mgrs/dl in the open group and from 2 to 1.1 mg/dl in laparoscopic group (p < 0.001). CONCLUSIONS: Renal grafts retrieved by laparoscopy presents a more evident ischemia reperfusion syndrome shown by a lower average RBF after unclamping and a significant deterioration of renal function during the first week after transplant. PMID- 18411633 TI - [Renal xenotransplant. Acute vascular rejection]. AB - INTRODUCTION AND OBJECTIVES: Organ transplant is nowadays a usual and succesful practice, although with limited application due to the lack of organs. Yearly thousands of patients get access to the waiting list and finally will death while they are waiting for an organ. In the U.S.A., 2005 waiting list for kidneys, heart, liver lung and pancreas was around 94.419. Number of transplants performed was 27.966 and died patients while waiting for an organ, 41.392 (1). Pig xenotransplant is one of the possibilities to ameliorate the lack of organs for transplant. Arrangement of pigs with different genetic modifications generated great expectatives on the use of these organs in clinics. Although preclinical experimental studies with kidneys reached prolonged survivals, these are really insufficient to go on with the clinical appliance. Hyperacute rejection produces destruction of the organ immediately. This problem could be pharmacologically precluded in xeno-transplant. However, acute rejection or vascular rejection usually produces the lost of the implant. New inmunosuppresive schedules delay significantly rejection, but not definitively. Xenotransplant as a therapeutic option introduces important scientific problems, as well as ethical and social. This paper reports a summary of our experience in renal xenotransplant and the management of acute rejection. MATERIAL AND METHODS: Twenty xenotransplants from transgenic pig (hDAF) as donor to babuine as receptor. Average weight of the animals ranged 11.4-75 kgrs and babuines 10-26 kg. Xenograft average weight ranged 39-160 grs. Implant was performed to aorta and cava. Four inmunosupressive schedules were used. RESULTS: Average survival was 7-9 days. Final Histological findings are described. Changes observed were secondary to acute tubular necrosis mixed with changes due to acute rejection. Three grafts were lost due to technical major problems. CONCLUSIONS: Although we have observed some promising results, xenotransplant is a very difficult problem to solve in the long-term. A lot of research is still needed-. PMID- 18411634 TI - [Experimental laparoscopic renal autograft as learning laparoscopic model]. AB - INTRODUCTION: Laparoscopic surgery requires a long training period of time in which the complexity of the training is increased. GOALS: The technique presented in this paper has been developed in order to find an experimental model that allows us to improve the learning of the vascular suture. Our main goal was to evaluate this technique as an experimental model for the vacular anastomosis, not to obtain a functional autotransplant. In this regard, here we summarize our experience during the first two cases performed. MATERIAL AND METHODS: Laparoscopic renal autotransplant was perfomed in female lab pigs weighing 15-20 kg. International Experimental Animal Care rules were accomplished. RESULTS: After sacrifizing the animals, only one case of vascular thrombosis was observed. The other cases showed normal arterial and venous flow. CONCLUSIONS: Experimental Renal Laparoscopic autotransplant constitutes a good surgical model. We are trying to implement the technique in the clinics in the next future. PMID- 18411635 TI - [Psychomotor and cognitive development of children born after intracytoplasmic sperm injection]. AB - OBJECTIVE: To analyse data on psychomotor and cognitive development of children born after intracytoplasmic sperm injection (ICSI). DESIGN: Open cross-section clinical study. SETTING: Institute for the Care of Mother and Child, Prague and Department of Paediatrics, Charles University, 2nd Medical School and University Hospital Motol, Prague. METHODS: In 133 children (75 boys and 58 girls) psychological examination was made at the age range 11 months - 8.5 years in the years 2004-2006. All children were born after intracytoplasmic sperm injection (ICSI). Psychomotor development of children aged from 11 months to 3.5 years was assessed using the Bayley Scales (BSID-II). In older children, Global Intelligence McCarthy Test was used. RESULTS: In our sample of ICSI-children, no significantly higher incidence of children delayed in mental (cognitive) as well as in motor development has been found as compared with the population norms. However, the results indicate a significantly lower average value of the Psychomotor Developmental Index (PDI) in the group of younger children as compared with the given norm (92.3 +/- 13.9 versus 100 +/- 15; p<0.01). In the group of older children, lower average value of the General Cognitive Index (GCI), as compared with corrected population norm has been found (105.1 +/- 14.7 versus 110 +/- 16; p<0.05). In the group of twins, a significantly higher number of mild developmental disorders was ascertained on the contrary in the group of singletons (64.7% versus 333%; p<0.01) in our older children conceived by ICSI. CONCLUSION: The results indicate only mild lowering of some performances in our ICSI-children: in motor domain in younger children, and in cognitive domain in older children. Children from multiple pregnancies are at greater developmental risk than singletons. PMID- 18411636 TI - [Parental attitudes and socio-emotional development in children born after ICSI]. AB - OBJECTIVE: To analyze parental attitudes and socio-emotional development of children conceived by ICSI. DESIGN: Open cross-sectional study. SETTING: Department of Clinical Psychology, Thomayer University Hospital, Prague and Department of Paediatrics, Charles University, 2nd Medical School, University Hospital-Motol, Prague. METHODS: 133 children (75 boys, 58 girls) conceived by ICSI, age ranged from 11 months to 8.5 years were psychologically assessed between the years 2004-2006. Children's behavior was evaluated by 4 rating scales during the assessment. Parents answered questionnaires concerning children's temperament, behavioral problems (TBC) and the parental attitudes questionnaire (PARQ). RESULTS: Children's behavior during the psychological assessment was rated mostly as very good or good, although the children were often less communicative. Most of the children have mixed or easy temperament, a difficult type of temperament didn't report any of the parents. Most of the parents didn't describe significant behavioral problems in their children, in particular there were very few externalizing difficulties (opposition, aggression), but in 29.5% of the sample, there were found some social or emotional difficulties. We found surprisingly high frequency of milder forms of autism spectrum disorders and another social problems (social and other anxiety disorders) in the sample, other psychopathology was rare. Parental attitudes had a tendency to grater involvement with the child and high affection in relation with him. CONCLUSION: Socio emotional development of ICSI children is good, although some have specific social difficulties, externalising problems were present only exceptionally. Parental attitudes toward ICSI children are positive, there is slight tendency to higher emotional involvement with the child. PMID- 18411637 TI - [Children born after intracytoplasmic sperm injection (ICSI): factors of infertility, pregnancy complications, neonatal period and neonatal characteristics]. AB - OBJECTIVE: To analyze the type of infertility, pregnancy and neonatal outcome in children conceived after intracytoplasmic sperm injection (ICSI children). DESIGN: Prospective open cross-sectional clinical study. SETTING: University hospital and private IVF unit. METHODS: Type of infertility, pregnancy complications, neonatal period and neonatal characteristics were evaluated in 135 newborns conceived after ICSI from singleton and twin pregnancies and compared to general population. RESULTS: The percentage of twins was significantly higher after ICSI compared to general population (31% versus 1.7%; p<0.001) as well as the percentage of caesarean section deliveries (31% versus 17.8%; p<0.001). Some complication in neonatal period was found in 21.5% ICSI newborns (18 out of 42 twins and 12 out of 93 singletons; p< 0.001). Some complication during the course of pregnancy was found in 50.9% ICSI children. CONCLUSIONS: no differences in gestational age, birth weight and birth length were found when ICSI and spontaneously conceined (sc) singletons and ICSI and SC twins were compared. However, complications during the course of pregnancy and in the neonatal period were more frequent in ICSI conceived children. PMID- 18411638 TI - [Birth defects, medical outcome and somatic development in children conceived after intracytoplasmic sperm injection (ICSI)]. AB - OBJECTIVE: To analyze the incidence of birth defects, medical outcome and somatic development in children conceived after intracytoplasmic sperm injection (ICSI). DESIGN: Prospective open cross-sectional clinical study. SETTING: University hospital and private IVF unit. METHODS: 135 Czech children (59 girls, 76 boys) from singleton and twin pregnancies conceived after ICSI (age 03-9.5 years; median 5.9) were assessed during the period 2004-2006. The incidence of birth defects, medical outcome and somatic development were evaluated and compared with data of general population and/or with control group matched for sex and age. RESULTS: Birth defects were found in 133% of ICSI children (compared to 4.6% in children after spontaneous conception; p<0.001). The general health of ICSI children did not differ significantly compared to general population. ICSI children required more surgery or hospitalization compared to general population data. There is high rate (69.6%) among ICSI children in the care of various specialised clinics. Body height and weight in ICSI children is in normal range and corresponds to their growth potential. Head circumference in ICSI children is larger compared to reference data (0.43 SD; p<0.001). CONCLUSIONS: No clinically important differences in somatic development between ICSI and general population of Czech children were found. Birth defects were more frequent in ICSI children. The overall general health in ICSI children seems satisfactory but ICSI children were more likely to need health care compared to general population. PMID- 18411639 TI - [The quality of breastfeeding preparation in the antenatal courses]. AB - OBJECTIVE: To evaluate the quality of breastfeeding preparation in the antenatal courses and to identify the other sources of breastfeeding information. DESIGN: Prospective study. SETTING: Thomayer's Hospital Prague. METHODS: Data collected from questionnaires presented on web pages of the Lactation League from 2005 to 2006 from 351 mothers were statistically analyzed. RESULTS: Only 52.8% of mothers were satisfied with the course content. Women who attended the antenatal preparation were more informed about the correct breastfeeding technique, the harmfulness bottle and pacifier usage, the disadvantages of the mother and newborn separation and alternative feeding methods than those who did not. The other sources of breastfeeding information were literature, magazines and internet. The health professional participation was only 9%. During pregnancy 82.9% of women did not pass the breast examination. CONCLUSIONS: It is necessary to improve the breastfeeding preparation quality of antenatal courses and to increase the health professional participation in breastfeeding information distribution. PMID- 18411640 TI - [Thyroid disease in the pregnancy and congenital malformations of newborns]. AB - THE AIM: The aim of this study was to compare thyroidal status of mothers who delivered newborns with morphological congenital malformations and mothers with healthy newborns. TYPE OF THE STUDY: One-time cross-sectional study. SETTING: 3rd internal clinic FN LP, Kosice. METHODS: We examined 43 mothers with newborns born with congenital malformation and 76 mothers with healthy newborns. The protocol included anamnesis, basic physical examination, ultrasonography of the thyroid, assessment of the mothers' plasma fT4, TSH and anti-TPO levels, and comparison of the birth weight and birth length of the newborns. The results were statistically processed by PC programs StatsDirect 2.6.2 and MS Excel 2000. RESULTS: The thyroid volume was significantly greater in mothers with newborns with congenital malformation as in the control group (p<0.001). The birth weight and birth length was significantly lower in newborns with malformations (p<0.0001 resp. p<0.001). We found no differences in the plasma levels of fT4, TSH and anti-TPO between these groups. CONCLUSION: Thyroid disease of the mother can contribute to the congenital malformations of the newborn although there was found no direct association between mothers' thyroid autoantibodies and creation of congenital malformations of the child, yet. PMID- 18411641 TI - [Thyroid disease in the pregnancy and hypotrophy of newborns]. AB - THE AIM: The aim of this study was to compare thyroidal status of mothers who delivered newborns with hypotrophy and mothers with healthy newborns. TYPE OF THE STUDY: One-time cross-sectional study. SETTING: 3rd internal clinic FN LP, Kosice. METHODS: We examined 56 mothers with hypotrophic newborns and 76 mothers with healthy newborns. The protocol included anamnesis, basic physical examination, ultrasonography of the thyroid, assessment of the mother's plasma fT4, TSH and anti-TPO levels, and comparison of the birth weight and birth length of the newborns. The results were statistically processed by PC programs StatsDirect 2.6.2 and MS Excel 2000. RESULTS: The thyroid volume was significantly greater in mothers with hypotrophic newborns as in the control group (p<0.001). The birth weight and birth length was significantly lower in hypotrophic newborns (p<0.0001 resp. p<0.0001). Mothers with hypotrophic newborns had significantly higher TSH levels (p<0.0001) and higher anti-TPO levels (p<0.0001) as mothers in the control group. We found no differences in the fT4 levels in these groups. CONCLUSION: Thyroid disease of the mother can contribute to the hypotrophy of the newborns. PMID- 18411642 TI - [Abdominal sacrocolpopexy--simple and still actual method in the treatment of vaginal vault prolapse?]. AB - OBJECTIVE: To evaluate the effectiveness and complications of abdominal sacrocolpopexy in the treatment of vaginal vault prolapse. DESIGN: Review article. SETTING: Obstetrics and Gynecology Department, Charles University 2nd Medical Faculty and Teaching Hospital Motol, Prague. METHODS: Review from biomedical database Medline from 1960 till 2006. The articles were found under the key words: vaginal vault prolaps, abdominal sacrocolpopexy, effectiveness, complications, urinary incontinence, female sexual dysfunctions. RESULTS: The effectiveness of surgery is getting near to 100 percent in the smaller groups between 11 and 21 women. The larger groups are presenting greater success rate variability from 85 to 97 percent. The studies with up to 2 years of follow up are giving representative results of the effectiveness of surgery. Prolapse recurrence and cystocele, rectocele, enterocele and stress urinary incontinence occurrance belong among the characteristic failure signs. Studies evaluating abdominal sacrocolpopexy argue low incidence of complications. Complications connected with surgery can be divided to intraoperative and early and late postoperative. Bleeding from injured medial sacral artery, cystotomy, enterotomy and ureterotomy are the most common intraoperative complications. Wound infection, ileus and urinary tract infection are the most often early postoperative complications. Stress urinary incontinence, anterior or posterior vaginal wall descent, recurrence of vaginal vault prolapse and mesh erosion through vaginal wall are characteristic late postoperative complications. CONCLUSIONS: Abdominal sacrocolpopexy can be a method of choice in the treatment of vaginal vault prolapse, mainly because of the low morbidity and high success rate. Polypropylene mesh is the prefered suspension structure but the accurate surgical technique is the most important for the abdominal sacrocolpopexy success and safety. PMID- 18411643 TI - [Ovarian tumor as a complication in diagnosis of the extraintestinal gastrointestinal stromal tumor--two case reports]. AB - OBJECTIVE: The aim of this study is to address difficulties in diagnosis of the extraintestinal gastrointestinal stromal tumor (GIST) when even in ovary is the tumor. One should ever think of the GIST in the case of peritoneal dissemination of the spindle/epithelioid cell tumor. In contrary, the late intraabdominal recurrence of primary both epithelial and non-epithelial tumor of genital tract can be undistinguishable by pure morphology. Complete clinical records and accurate operation field topography are both of great value. Documenting this phenomenon, presented is both late intraabdominal dissemination of primary uterine endometrial stromal sarcoma and massive ovarian involvement by extraintestinal GIST. DESIGN: Two case reports. SETTING: Biopsy Lab s.r.o. and Sikl's Department of Pathology, Charles University and Faculty Hospital, Pilsen. METHODS: The two casuistic reports are covered in detail including broad immunohistochemistry (LSAB+, Dako) and genetics (ABI Prism 310, PE/Applied Biosystems). RESULTS: In 75-year-old woman, 5 cm tumor in maximum diameter of the mesentery was diagnosed as an extraintestinal GIST. Multiple peritoneal recurrence occurred ten years later and the tumor has been reclassified as a disseminated endometroid stromal sarcoma. Ultimately, comprehensive search of clinical files confirmed hysterectomy and adnexotomy in 1973. The second case describes 49-year-old woman operated for tumorous obliteration of the small pelvis where a tumor 9 cm in maximum diameter was found. Initially, predominant epitheloid morphology of the tumor led to the diagnosis of primary epithelial malignancy, sex-cord stromal tumor or metastatic melanoma of the ovary. Finally, the help of immunohistochemistry and molecular biology rendered the tumor as an extraintestinal GIST, ovarian primary not excluded. PMID- 18411644 TI - [Caudal regression syndrome--two case reports]. AB - OBJECTIVE: The authors demonstrate two cases of caudal regression syndrome (CRS), a rare malformative syndrom, seen mainly in cases of maternal diabetes with poor metabolic control. DESIGN: Case report. SETTING: Department of Obstetrics and Gynecology, Department of Medicine Regional Hospital Pardubice. CASES: The caudal regression syndrome (CRS) was revealed in two women with praegestational diabetes. The diagnosis was made at 18 and 20 weeks. The characteristic ultrasound findings include abrupt interruption of the spine and abnormal position of the lower limbs. The femur bones are fixed in a "V" pattern, giving a typical "Buddha's poise". A complete examination must be conducted for possible urinary and intestinal malformations. The mechanism leading to malformation is discussed in the article. To prevent pregnancy at the time of bad controlled diabetes is the only way to minimaze the risk of producing a congenitally malformed baby including caudal regression syndrom in the population of diabetic mothers. Family planning and supervision by the specialists is always advisable. CONCLUSION: Early diagnosis of CRS is possible using vaginal ultrasound. Emphasis is placed on the association of abrupt disruption of dorsal or lumbar spine and abnormal images of the lower limbs fixed in a,,V" formation, which is characteristic sign of CRS. PMID- 18411645 TI - [Intraspecific taxonomic differentiation in ticks (Acari: Ixodidae) in the view of the morphological conception of species]. AB - The criterions for two most used intraspecific taxonomic ranks of ticks- subspecies and morphotype, have been formulated on the basis of the study of morphological variation in the distribution range of all active ontogenetic stages of 11 polymorphic species. All these species are vectors of transmissible diseases. They have vast distribution ranges and different types of host-parasite relationships. Subspecies have the complexes of visual morphological differences expressed in one or both sexes of mature ticks more limited, than those of related species. At immature stages differences of subspecies consist more often in morphometric characters and can be established by the methods of mathematical statistics only. Morphotypes, as a rule, differ at each corresponding stage by "their own" complexes of morphometric characters. All differential parameters of studied morphotypes are overlapped, but have statistically significant differences (by the Student's test). The concrete variations of differentiation by subspecies and morphotypes have been considered. The historical factors of intraspecific differentiation have been reconstructed for each species. PMID- 18411646 TI - [Morphological adaptations of acariform mites (Acari: Acariformes) to permanent parasitism on mammals]. AB - The external morphological adaptations to parasitism in acariform mites (Acari: Acariformes), permanently parasiting mammals, are briefly summated and analyzed. According to several external morphological criteria (structures of gnathosoma, idiosoma, setation, legs and life cycle), the following six morphoecotypes were established: skin mites (i)-- Cheyletidae, Chirorhynchobiidae, Lobalgidae, Myobiidae, Myocoptidae (the most part), Rhyncoptidae, Psoroptidae; fur mites (ii) -Atopomelidae, Clirodiscidae, Listrophoridae, Myocoptidae (Trichoecius only); skin burrowing mites (iii)--Sarcoptidae; intradermal mites (iv) - sorergatidae and Demodicidae; interstitial mites (v) - pimyodicidae; respiratory mites (vi) - reynetidae, Gastronyssidae, Lemurnyssidae, Pneumocoptidae. In the case of prostigmatic mites, the detailed reconstruction of the origin and evolution of "parasitic" morphoecotypes is possible due to the tentative phylogenetic hypotheses, which were proposed for the infraorder Eleutherengon, a, including the most part of the permanent mammalian parasites among prostigmatic mites (Kethley in Norton, 1993; Bochkov, 2002). The parasitism of Speleognathinae (Ereynetidae) in the mammalian respiratory tract arose independently of the other prostigmats. It is quite possible that these mites switched on mammals from birds, because they are more widely represented on these hosts than on mammals. The prostigmatic parasitism on mammalian skin seems to be originated independently in myobiids, in the five cheyletid tribes, Cheyletiellini, Niheliini, and Teinocheylini, Chelonotini, Cheyletini, and, probably, in a cheyletoid ansector of the sister families Psorergatidae-Demodicidae (Bochkov, Fain, 2001; Bochkov, 2002). Demodicids and psorergatids developed adaptations to parasitism in the skin gland ducts and directly in the epithelial level, respectively in the process of the subsequent specialization. Mites of the family Epimyodicidae belong to the phylogenetic line independent of other cheyletoids. These mites possess the separate chelicerae and, therefore, can not be included to the superfamily Cheyletoidea. It is not quite clear whether they were skin parasites initially or they directly switched to parasitism from the predation. The phylogeny of sarcoptoid mites (Psoroptidia: Sarcoptoidea) is not developed, however, some hypotheses about origin and the following evolution of their morphoecotypes can be proposed. We belive that astigmatic mites inhabiting the mammalian respiratory tract transferred to parasitism independently of other sarcoptoids. The idiosoma of these mites is not so much flattened dorso-ventrally and has proportions which are similar to hose of free-living astigmatids. Moreover, in the most archaic species, the legs are not shortened or thickened as in the most parasites. The disappearance of many morphological structures in these mites, probably, happened parallely with some other sarcoptoids due to their parasitic mode of life. The skin inhabiting sarcoptoids belong to the "basic" morphoecotype, and all other sarcoptoid morphoecotypes, excluding respiratory mites, are derived from it. Some mites of this morphoecotype live on the concave surfaces of the widened spine-like hairs of the rodents belonging to the family Echimyidae (mites of the subfamily Echimytricalginae), in the mammalian ears (some Psoroptidae) or partially sink into the hair follicles (Rhynocoptidae). Finally, mites of the family Chirorhynchobiidae live on the bat wing edges attaching to them by their "ixodid-like" gnathosoma. The fur sarcoptoids, probably, originated from the skin mites. This morphoecotype is divided onto two subtypes: mites with the dorso-ventrally flattened idiosoma (subtype I) and mites with the teretial idiosoma (subtype II). Each "fur-mite" family includes mites of the both subtypes. All mites of the first subtype belong to the early derivative lineages in their families. Among listrophorids such early derivative lineage is represented by the subfamily Aplodontochirinae (Bochkov, OConnor, 2006), and among Chirodiscidae--by mites of the subfamilies Chirodiscinae, Schizocoptinae, and Lemuroeciinae. Among the "fur" astigmatid families, the family Atopomelidae. probably, is the most archaic, and the most part of atopomelids belongs to the first subtype. However there are several more specialized atopomelid genera belonging to the second subtype, Atopomelus, Dasyurochirus, Lemuroptes, Murichirus, Metachiroecius etc. We believe that mites of the first subtype are represented by the "intermediate" forms between skin mites and mites of the second subtype. Some skin sarcoptoids transferred from skin parasitism to burrowing of the host skin (Sarcoptidae). The established morphoecotypes are partially corresponding to some morphoecotypes established by Mironov (1987) for feather mites. Our morphoecotypes of skin and skin burrowing mites perfectly correspond to Mironov's epidermoptoid and knemidocoptoid morphoecotypes, respectively. The proctophylloid morphoecotype (mites living on the wing feathers), which is the most widely represented within feather mites, has an analogy among mammalian mites - the subfamily Echimytricalginae. The analgoid (mites living in the down feathers) and dermoglyphoid (mites living in the feather quills) morphoecotypes have no analogues among mammalian mites for the obvious reasons. It is interesting why some microhabitats on the host body are not still occupied by prostigmatic or astigmatic mites. We believe that the nutrition is the main limitative factor here. The parasitic prostigmates evolved from predators and, therefore, feed on content of the living cells. They need the direct contact with the live tissues of the host and they belong, therefore, to the morphoecotypes represented by the respiratory, skin, gland duct, intradermal, and interstitial mites. Whereas, the most part of the skin inhabiting astigmats feed on the dead epithelial scales. For this reason these mites, so easily colonized fur of their hosts and feed on the hair grease there. On the other hand, some sarcoptoids transferred to the true parasitism and feed on the cambial cells of the skin epithelium. More over we do not know exactly about nutrition of rhyncoptids yet. PMID- 18411647 TI - [Species composition of mosquitoes (Diptera, Culicidae) and possibility of the West Nile virus natural foci formation in the South of Western Siberia]. AB - In 2004 June-July collections of mosquito adults and small mammals were carried out in two areas of Novosibirsk Region (forest-steppe and steppe zones), where the West Nile virus (WNV) was for the first time recorded in birds with different migration status in 2002-2004. Seventeen species of mosquitoes were found; significant changes in their species composition and abundance, as compared with latest faunistic studies made in the sixties-seventies of the last century, are revealed. WNV markers (antigen, RNA) are found in small mammals; highly sensitive to the WNV replication mosquito species are also found. These facts allow supposing a possibility of the formation of stable West Nile virus natural foci in the South of Western Siberia, under conditions of forest-steppe and steppe zones. PMID- 18411648 TI - [Mosquitoes (Diptera: Culicidae) of anthropogenic territories in the North of Tomsk Region (by an example of the Strezhevoy town)]. AB - An ecological-faunistic analysis of mosquitoes has been carried out in the Strezhevoy town (Tomsk Region). Thirty-one species of mosquitoes have been recorded in the territory examined. The population peak of mosquitoes is recorded in the 1-2 ten-days of July. Characteristics of the biotopic distribution and daily activity of the dominant mosquito species are revealed. Limiting factors of daily activity of mosquitoes in the investigated territory are characterized. PMID- 18411649 TI - [Main evolution lines of plant parasitic nematodes of the order Aphelenchida siddiqi, 1980]. AB - Phylogenic models for each aphelenchid family and phylogeny of the order Aphelenchida as a whole were developed on the base of detailed comparative morphological and bionomical analysis of the order. Bionomical and morphological characters having a phylogenetic significance were selected. Classification proposed by Hunt, 1993 was used as the starting-point of the study. Life cycles and their evolution in Aphelenchida were analyzed on the base of phylogenetic trees. It is concluded, that aphelenchid ancestors combined mycophagy, plant parasitic, and partly predaceous feeding. Relations of the primitive Aphelenchida with their symbionts developed from the spots of the fungal organic matter decomposition in the "nema- tode-fungi" associations, followed by a transition to the temporary endoparasitic habit omitting ectoparasitism. With a complication of the nematodes' life cycles, the insect vector (detritophagous or pollinator) transformed into the real insect host of the parasitic nematode in the 2-host life cycle (with the plant and insect hosts) or in the obligate 1-host entomoparasitic life cycle of the aphelenchid nematodes. Specialization of the aphelenchid life cycles to insect vectors followed two main ways. In the first way, the resistant to unfavorable environmental conditions nematode juveniles, known already for the primitive aphelenchids transformed into dispersal juveniles, and later into parasitic juveniles. In the second evolution line the dispersal function were laid on inseminated but non-gravid (not egg-producing) females. Both above-mentioned trends of parasitic specialization were arisen independently in different phylogenetic lines of the Aphelenchida. In each line of the parasitic development in different nematode families, the highly specialized ectoparasites, as well as endoparasites on insects, were formed. In the evolution of life cycle of parasitic nematodes, a tendency to decrease the body size took place. The function of dispersion shifted to more junior juvenile stage (the first line of specialization), or body sizes of non-gravid females and males copulated with the latter become smaller (second specialization line, till the development of dwarf males and location of the males and small inseminated non-gravid females in the uterus of gravid nematode female). The hypothetic fundamental model of the parasitic cycles' specialization in the order Aphelenchida was developed, basing on the comparison of known life cycles in different phylogenetic lines within aphelenchid families. The conception of the geographic origin and historic dispersal of the order Aphelenchida was proposed. The origin of the superfamily Aphelenchoidoidea and order Aphelenchida as a whole probably took place in eastern areas of Gondwana (parts of which are recently Hindustan, Indo-Malaya, Australia and Antarctica), presumably in the Devonian period. When the Gondwana and Laurasia paleocontinents were joined into Pangea in Carbon period, aphelenchids dispersed in the Laurasian part of Pangea. Endemism of the advanced entomophilic ectoparasitic Acugutturidae indicates on the secondary hotbed of speciation in Caribbean area. Development of the anhydrobiotic adaptations in the Aphelenchida promoted their successful invasion in the cold regions of Holarctic. Another important adaptations was the transformation of the initially resistant nematode life cycle phase into the dispersal phases vectored by insects. PMID- 18411650 TI - [Reproduction of the trematode Echinostoma caproni parthenites (Digenea: Echinostomatidae)]. AB - Dynamics of the reproduction in the trematode Echinostoma caproni parthenites (Echinostomatidae) was observed. Early laying and maturation of the generative cells are for the first time shown to be characteristic for all parthenogenetic generations. Really the process of reproduction had been finishing to the beginning of the generating of new age by parthenites. Mother sporocysts, as well as redia of different generations, in fact stop producing new generative cells with the beginning of the generating of new age, and assume the function of a brood pouch. This feature was considered previously as peculiar mainly to mother sporocysts. Data on the autotomy of the anterior body end in mother sporocysts are verified. In our opinion, these data are an evidence of an early manifestation of the evolutionary trend to the morpho-functional regress and disintegration of the parasitic stage of mother sporocyst. PMID- 18411651 TI - [Successional features of the dynamics of abundance and population structure of the cestode Proteocephalus longicollis (Zeder, 1800) (Cestoda: Proteocephalidae)]. AB - Dynamics of abundance and changes of populations structure were investigated in a host-specific parasite of salmonids, cestode Proteocephalus longicollis from the vendace Coregonus albula L. of the Syamozero Lake during the period of more than forty years. A relative stability of the values of infestation indices independently of significant changes in the abundance and population structure of the vendace is established. Changes in the distribution of abundance and population structure of P. longicollis are shown to be connected with the change of the water body's status resulting from eutrophication. These changes can be considered as reliable indices of the succession in freshwater bodies. PMID- 18411652 TI - [The role of resident and circulating stem cells in physiological and reparative regeneration]. AB - Complete or partial restoration of the number of parenchymal cells is a vital part of the mechanism of maintaining tissue homeostasis and post-injury regeneration. It includes proliferation and differentiation of the resident stem cells, homing and differentiation of circulating stem cells, and, sometimes, transient dedifferentiation and amplification of the local parenchyma. Bone marrow serves as the principal storage site of the circulating stem cells. Several possible ways of circulating stem cells participation in tissue regeneration are discussed including trans-differentiation and fusion with resident cells. There is a theory of a common pool of circulating pluripotent stem cells present in the bone marrow and some other tissues and an alternative theory of several pools of tissue-specific circulating stem cells. The data obtained in this field of research are the starting point for development of cell based technologies aimed at therapy of degenerative diseases and traumas. PMID- 18411653 TI - [Immune pathology in pulmonary tuberculosis]. AB - The data of modem literature and the results of original investigations on mechanisms of immunopathological alterations in tuberculosis infection are presented. The role of cellular and humoral parts of immunity in pathogenesis of pulmonary tuberculosis is discussed and cytokine-mediated mechanisms of disorders in a specific immune response are analysed. PMID- 18411654 TI - [Thromboerythrocytic immunosuppression and its correction by cell membrane stabilizers during intensive exercise]. AB - Erythrocytes possessing immunosuppressive properties appear in the blood of animals after intensive physical exercise. These properties of erythrocytes arise during their interaction with platelets and LDL+VLDL fraction. Essenciale increases erythrocyte resistance to the action of platelets modified by lipoproteins while mildronate destroys interaction of lipoproteins and platelets. PMID- 18411655 TI - [Ganoderma lucidum extract in cardiac diastolic dysfunction and irreversible cardiomyocytic damage in ischemia and reperfusion of the isolated heart]. AB - We used a model of total 45-min ischemia and 30-min reperfusion of isolated rat heart by the Langendorf technique. The course administration (15 days) of Gonoderma lucidum extract attenuated reperfusion contracture and decreased creatine kinase levels in the venous effluent from rat isolated heart during reperfusion. However, the extract did not prevent a reperfusion decrease in pressure developed by the left ventricle, reduction in the heart rate, contraction and relaxation rates. The extract had no effect on the incidence of ventricular arrhythmia. We believe that Ganoderma lucidum extract is a drug which attenuates diastolic dysfunction and prevents irreversible cardiomyocyte damage during ischemia and heart reperfusion. PMID- 18411656 TI - [A hepatoprotective effect of medicinal herbal tea hexacholefit in ethanol induced liver damage]. PMID- 18411657 TI - [Introduction of new technologies in teaching basic and clinical pathophysiology]. AB - The article contains data on teaching basic and clinical pathophysiology in Omsk state medical academy. Teaching process includes tests of the first and second level of difficulty, solving situational profession-oriented problems, quizzes, demonstration of patients and pathophysiological analysis of clinical syndromes, case reporting and recording. PMID- 18411658 TI - [Antonin Nikolaevich Filatov (1902-1974)]. PMID- 18411659 TI - [Antithrombotic therapy in operations on extracranial arteries]. AB - Application of Clopidogrel in the postoperative period in patients after surgery on extracranial arteries was analyzed. The preparation was used in 56 patients. Uninterrupted intake of the drug took place during 5 to 12 months, 7+/-1.6 months on average. Stomach and duodenal ulcer was diagnosed in 11 patients (20%), 3 patients (5%) had chronic pancreatitis with periodical exacerbations and 2 patients (4%) had erythrocytemia. One bleeding requiring revision of the wound took place in the nearest postoperative period. During the whole period of observation there were no thrombotic or thromboembolic complication against the background of using plavix. In none of the patients granulocytopenia was revealed with continuous intake of plavix. An ultrasonic investigation of the operated arteries in the remote postoperative period failed to reveal hemodynamically significant restenoses of carotid arteries. PMID- 18411660 TI - [Distal reconstructions with the autovein in situ for critical ischemia of lower extremities in elderly and senile patients]. AB - The work was devoted to the problem of early and late thromboses of transplants at femoro-popliteal-tibial shunting. One of the main causes of frequent thromboses of the shunts was soundly shown to be high peripheral vascular resistance. The investigation has shown efficiency of using collaterals of the large subcutaneous vein as collectors of unloading the shunt in order to decrease the peripheral vascular resistance in femorodistal reconstruction with the autovein in the in situ position. In a year the cumulative passability of the shunt in the main group patients treated by the method of unloading the shunt with the help of non-ligated autovein collaterals made up 75.1% compared to 69.3% in patients of the group of comparison, and within 5 years 53.8% and 45.2% respectively. The index of safety of the extremity in a year was 81.5% in the main group patients, and 73.2% in control group patients, in 5 years 64% and 46.5% that demonstrated advantages of the method of leaving the functioning tributaries of the large subcutaneous vein intact after reconstruction. PMID- 18411661 TI - [Functional-anatomical prerequisites of revascularization of the femoro-popliteal arterial segment]. AB - Improved results of surgical treatment of patients with critical ischemia of lower extremities can be obtained by using the collateral bed. The condition of the profound femoral artery and the popliteal artery, especially in the zone of its trifurcation, is of the greatest significance for the decision on the volume of surgery. The carrying capacity of the collateral bed of the profound femoral artery in occlusion of the femoral artery was on average 284 ml/min. A positive prognostic criterion of recovered circulation in the extremity using the profound femoral artery is preservation of the patent trifurcation of the popliteal artery and/or the anterior tibial artery. The blood flow volume along the profound femoral artery under the given functional-anatomical conditions should be not less than 150 ml/min. Semiclosed loop endarterectomy of the superficial femoral and popliteal arteries with multilevel lesions of the lower extremity arteries allows the main zones of the collateral bed of the femoro-popliteal-tibial segment to be included in the blood flow. PMID- 18411662 TI - [Sclerotherapy of sapheno-femoral reflux of blood in patients with initial stages of varicose disease of the lower extremity veins]. AB - The results of ultrasonic investigation and the following phlebosclerosing treatment of an incompetent saphenofemoral anastomosis were analyzed in two groups of patients with initial stages of varicose disease of the lower extremity veins. The first group included 48 patients treated by injection-sclerosing therapy by the technology of "empty vein". The second group consisted of 82 patients treated by catheter sclerotherapy of the sapheno-femoral anastomosis. In the first group the treatment was effective in 73.1%, in the second group - in 91.5%. The terminal hemodynamic criteria of performing the injectional and catheter sclerotherapy were determined in elimination of high sapheno-femoral reflux of blood. PMID- 18411663 TI - [Application of cryogenic stimulation in treatment of chronic wounds]. AB - The authors have studied alterations occurring both in the ultrastructure of the cell matrix and in the microcirculatory bed of the chronic wound after local exposure to cryoagent. The up-to-date effective methods including laser Doppler flowmetry were used followed by correct statistical processing of the data obtained. The cryogenic stimulation of the wound was shown to result in considerably improved perfusion of the microcirculatory bed, epithelization and remodeling of the scar. It allowed transformation of a chronic process into acute and thus led to considerably accelerated process of regeneration. The developed method of cryogenic treatment of the chronic wound was used in 35 patients, allowed quicker healing of the chronic wounds and made ambulatory treatment of the patients 3 weeks shorter. PMID- 18411664 TI - [Laparoscopic adjustable gastric banding in treatment of patients with obesity]. AB - Laparoscopic adjustable gastric banding (LAGB) has been fulfilled in 2958 patients in the period since 1998 through 2006 for treatment of different forms of obesity. The age of the patients was from 16 to 60 years (mean age 38 years). There were 80% of women (2366) and 20% of men (592). Mean preoperative body mass index was from 35 to 54 kg/m2, i.e 43+/-3.8 kg/m2, fluctuating from 35 kg/m2 to 54 kg/m2. The observation of 2485 (84%) patients operated in the terms in question showed the influence of LAGB on metabolic derangement, arterial pressure. Statistical data on possible early and late postoperative complications are presented. The operation of LAGB is one of minimally invasive methods for adjustable decreasing the excess body weight and its effectiveness exceeds other methods of restrictive interventions on the stomach, has a number of advantages compared with other bariatric interventions, it is relatively safe and completely reversible bariatric operation with all advantages of minimally invasive techniques. PMID- 18411665 TI - [Use of gastric balloon in treatment of patients with obesity]. AB - Gastric ballons (GB) were introduced in 37 patients (34 women and 3 men) aged from 18 through 57 years (mean age 35.2 years) for treatment of different forms of excess body weight and obesity. Mean index of the body mass at the moment of introducing the balloon was 102.3+/-8 kg varying within the range of 80 to 178 kg. The body mass index was on the average 37.9 kg/m2 (29-63). Possible complications and clinical manifestations after introduction of GB are considered. An analysis of the results of treatment and of the nearest and long term results shows that the body mass became on the average 78.9+/-5 kg (62-138), the body mass index decreased on the average to 29+/-1.6 kg/m2 (22-46). Maximum body weight loss was 48 kg. Using the gastric balloon is considered to be a minimally invasive technique directed to decreasing the excess body mass and its effectiveness to be higher than other methods of conservative therapy. The application of GB substantially extends the chances to help patients with obesity with the minimal risk for health and life. PMID- 18411666 TI - [Primary jejunogastroplasty with end-loop gastroenteroanastomosis and inclusion of the duodenum in distal resection of the stomach (surgical technology and functional results)]. AB - Technical aspects, clinical and functional results of using an original method of primary jejunogastroplasty (JGP) in resection of the stomach were considered. Complex examination of 57 patients (25 patients of the main group, and 32 controls) has shown that the proposed technology of primary JGP can reliably prevent gastroesophageal and duodeno-enterogastic reflux, creates conditions for the portion-rhythmical evacuation of food from the gastric stump. It is also characterized by a less, as compared with the controls, incidence and degree of alterations in the mucosa and better quality of life of the patients after operation. The results obtained show that primary JGP is an effective method for prevention of postresectional pathological syndromes. PMID- 18411667 TI - [Treatment of metaphyseal fractures of shin bones by the method of blocking osteosynthesis]. AB - The proposed method of reposition and polyaxial stabilization of fragments for intramedullary meallosynthesis of fractures of long tubular bones allows blocking osteosynthesis to be successfully used in treatment of complex metaphyseal fractures of shin bones. It results in strong fixation of the fragments, makes it possible to successfully eliminate residual deformities after introduction of the nail and to avoid the development of them in future under the influence of loading. The method provides early functioning of the interfacing joints, early axial loading, shorter period of disability, the absence of external immobilization. PMID- 18411668 TI - [Prophylactics of pyo-inflammatory complications in the wire area during treatment by the method of transosseous compressive-distractive osteosynthesis with probiotic "Sporobacterin liquid"]. AB - Prophylactics of surgical infections is one of the principal problems in using any surgical method, the method of transosseous osteosynthesis included. Preventive treatment is considered to be one of possible ways to decrease the number of pyo-inflammatory complications. However, unjustified antibiotic therapy gives a negative effect and is often followed by side reactions and complications. This experimental investigation presents grounds for using the method of prophylactics of pyo-inflammatory complications in the area of wires in treatment by the method of extrafocal compressive-distractive osteosynthesis with a new generation probiotic "Sporobacterin liquid". PMID- 18411669 TI - [European consensus on the management of patients with differentiated carcinoma of the thyroid from follicular epithelium]. PMID- 18411670 TI - [Method of treatment of combined injuries of the liver and spleen in children]. AB - Since 1980 operations for combined injuries of the liver and spleen have been made on 57 children aged from 5 through 16 years. Autografting of splenic tissue into the liver wound was performed in 17 (29.8%) patients using an original technique developed in experiments. The novelty of the suggested technique is that autografting of splenic tissue into the liver wound was followed by fixing the fragments with Pi-like stitches through the spleen transplant capsule. It ensures the impermeability of the liver wound, adequate hemostasis without applying artificial hemostatic materials and helps to avoid alloplastic materials for additional drainage of the subhepatic space, provides a direct physiological contact between the splenoid and liver tissue that is effective for prevention of hyposplenism following spleenectomy. PMID- 18411671 TI - [The use of magnetic resonance imaging in diagnostics of injury of the knee joint meniscuses]. AB - We analyzed the clinical symptoms, mechanisms of damage of the meniscuses in 286 children with special attention to diagnostic difficulties and causes of tardy hospitalization. MRI was performed in 286 patients for the diagnosis of injuries of meniscuses. In 208 children the diagnosis was made by arthroscopy. A statistical analysis has shown that the sensitivity of MRI was 89.1%, the specificity was 95.1%. PMID- 18411672 TI - [Postoperative anesthetization by the method of transcutaneous electroneuroanalgesia]. AB - The article is devoted to effectiveness of tanscutaneous electroneuroanalgesia using apparatus "Romitron 14T" for postoperative anesthetization in patients after not very traumatic operations. The efficiency of anesthetization was estimated by the visual-analogue scale, scale of sedation, changes in indices of hemodynamics, respiration and content of glucose in blood. It has been established that transcutaneous electroneuroanalgesia is an effective method for elimination of postoperative pain and can be used for complete substitution of medicinal kinds of postoperative anesthetization after not very traumatic operations. PMID- 18411673 TI - [Prophylactics of postpuncture syndrome after central neuroaxial blockades]. AB - The article presents data of an analysis of anesthesiologic maintenance of 3410 cesarean sections under conditions of spinal and epidural anesthesia. A detailed examination of 378 women has shown that frequency of the development of postinjection syndrome depends on the area of the defect of dura mater and the volume of lost cerebrospinal fluid. Postinjection syndrome can be prevented by using special needles leaving a less defect in dura mater and by introduction of 10-15 ml of sterile isotonic solution of sodium chloride into the epidural space. PMID- 18411674 TI - [Method of surgical treatment of stenosing ligamentitis of fingers]. AB - The authors present results of surgical treatment of patients with stenosing ligamentitis of fingers using different methods. A new method of operative treatment is described. The strategy of decision on this method is grounded, its practicability and results of treatment. PMID- 18411675 TI - [Method of surgical treatment of ganglion of the wrist]. AB - Comparative characteristic of the methods of treatment of patients with ganglion of the wrist has been carried out at the City Center of ambulatory surgery of the wrist since 1997 through 2006. A new method of surgical treatment is proposed which has been approved on 97 patients. No recurrences were noted during 5 years of follow-up. PMID- 18411676 TI - [On the choice of the material in surgical treatment of rectocele]. AB - An investigation of the physical (capillaries, elasticity, strength, traumaticity) and implantation properties of modem synthetic absorbable suture material "Maxon 1", "Polysorb 1", "Vicryl 1" has shown considerable advantages of the monofilic thread "Maxon 1". For surgical treatment of anterior rectocele of the III degree (anterior levatoroplasty) this thread was used in 114 patients with good direct, nearest and long-term results. PMID- 18411677 TI - [Diseases of veins in the elderly: stepwise treatment and rehabilitation]. AB - A specially developed card including 147 parameters was used in examination of 118 elderly phlebological patients. Pathology of the veins occurs in the elderly 5.4 times more often than in subjects younger than 60. A six-step system was developed for treatment of such patients: primary step, day hospital, specialized vascular and surgical unit, rehabilitation center, health resort treatment, dispensary examinations. Laboratory investigation of thrombophilic conditions, ultrasonic angioscanning and using minimally invasive methods of operative treatment should be introduced in the laboratory practice for the improvement of quality of life and prevention of complications. PMID- 18411678 TI - [Choice of operation for treatment of patients with pilonidal sinus]. AB - An analysis of early postoperative complications and recurrences of the disease was made in 136 patients with epithelial coccygeal canal. A simple method of suturing the wounds was used in 96 patients, in 40 patients Z-plasty was used. The results were estimated by the degree of the pathological process and the individual features of the structure of the sacro-coccygeal area. The results did not differ in patients with simple forms of the disease. In patients with complex form of the coccygeal canal wound abscesses were noted in 9 (34.6%) patients with simple stitching and in 2 (8%) after using Z-plasty. Recurrent diseases appeared in 6 (23.1%) patients after traditional methods of surgery and in 1 (4%) patients after Z-plasty. PMID- 18411679 TI - [Programmed sanation laparoscopy in treatment of peritonitis]. AB - An analysis of experiences with endovideosurgical sanation of the abdominal cavity was analyzed in 168 patients with diffuse peritonitis which complicated the course of different acute diseases. Relaparoscopy might be programmed and "on request". Programmed laparoscopy allowed adequate sanation of the abdominal cavity in most cases. It was impossible in 11.3% of cases associated with a pronounced commissural process and uncared peritonitis with formed pyogenic abscesses in the abdominal cavity. The indications and contraindications to laparoscopic sanation of the abdominal cavity are described. PMID- 18411680 TI - [Surgical treatment of nephrotuberculosis in the present-day epidemiological situation]. AB - The article presents data on the structure of operative interventions and assessment of the role of an early kidney decompression in management of renal tuberculosis complicated by urethritis. It was found that such eradicative techniques as nephrectomy and nephruretherectomy still prevail. Early drainage of the kidney for its decompression allows preservation of the kidney and following reconstructive surgery in 70.6% of cases. The number of early and later complications considerably decreased. PMID- 18411681 TI - [Long-term results of treatment of patients with liver echinococcosis operated by a traditional method and by using high temperature (plasma) technologies]. AB - A comparative analysis of surgical treatment of 160 patients with liver echinococcosis treated by traditional surgical methods and with using plasma technologies followed by a complex assessment of the long-term results has reliably confirmed advantages of operations using the plasma flow. They were followed by less incidence of postoperative complications and recurrences of the disease. It was also confirmed by high quality of life of the contingent observed that was estimated by a specially developed questionnaire. PMID- 18411682 TI - [Cases of pyogenic iliopsoas abscesses in military men]. AB - Four cases of pyogenic inflammation of the iliolumbar muscle were analyzed and 3 clinical cases of this disease in military men are described. A rare case of abscess of the iliolumbar muscle complicated by secondary appendicitis is also described. A hematogenic way of infection against the background of microtraumas and hematomas with m. Iliopsoas was the main one in the patients observed that was due to specificities of military service: permanent physical activity, pustular diseases of the lower extremities. The main clinical symptom of the disease is the symptom of the iliolumbar muscle. Ultrasonic scanning is the most informative method of diagnosis among additional methods. Operative treatment of the piogenic iliopsoas abscess is the only method of treatment, the open operative interventions by the retroperitoneal access being preferred. Antibacterial therapy in the postoperative period is based on the results of inoculation of the purulent discharge from the wound. Broad spectrum antibiotics must be used due to possible anaerobic character of the purulent inflammation. PMID- 18411683 TI - [Favorable outcome of surgical treatment of an artificial rupture of the trachea]. PMID- 18411684 TI - [Successful treatment of the trachea injured during intubation]. PMID- 18411685 TI - [A case of successful treatment of odontogenic mediastinitis complicated by bilateral empyema of the pleura using videothoracoscopy]. PMID- 18411686 TI - [Fournier's gangrene]. PMID- 18411687 TI - [Long-term results of arterial switch operation]. AB - Surgical results and late outcome in 202 patients who had undergone arterial switch operation from 1984 to 1997 were investigated. Actuarial survival was 90.6% at 10 years and 90.0% at 20 years. Fifty-two patients (25.7%) underwent reoperation for pulmonary stenosis and 7 patients (3.5%) had aortic valve replacement. Freedom from re-intervention was 71.9% at 10 years and 60.4% at 20 years. Using xeno-pericardial patch for pulmonary reconstruction was strong predictor for postoperative pulmonary stenosis. Coronary ischemic event was rare but some patients showed electorocardiogram (ECG) change on exercise and hypoplastic left coronary artery. Cardiopulmonary function was almost normal in long term survivors. PMID- 18411688 TI - [Arterial switch operation: late results and a future perspective]. AB - Intermediate results of the arterial switch operation for variety of cardiac anomaly with the transposed great arteries were studied to elucidate late results and problems. Seventy-five survivors of the arterial switch operation at our institute between 1991 and January 2007 were studied retrospectively. Mean follow up time was 7.5 years. There were 2 late deaths, 3 reoperations, and 9 cases of catheter intervention, resulting in actuarial survival of 97%, and event-free rate of 83% at 16 years. Seventy patients (93%) were medication free and had no restriction on physical activities. Moderate pulmonary stenosis in 2 patients and mild pulmonary stenosis in 14 were observed. Moderate aortic regurgitation in 2 and mild aortic regurgitation in 20 were observed. Risk factors for right ventricular outflow obstruction were a diagnosis of double outlet right ventricle (DORV), association with aortic arch obstruction, and prior pulmonary artery banding (PAB). Risk factors for branch pulmonary artery stenosis (PS) were not clear; however, scar formation with use of xenograft patch and in-fold formation by redundant autologous pericardial patch for closure of the defects in the old aortic root seemed to be related with the stenosis. Risk factors for aortic regurgitation were age at operation and prior PAB. Early primary arterial switch operation may resolve these problems. PMID- 18411689 TI - [What is necessary for improvement of arterial switch operation?]. AB - We report long-term results of the arterial switch operation (ASO) and postoperative complications related to operative procedure. Between 1998 and 2007, ASO were performed in 42 patients [transposition of the great arteries (TGA) with intact ventricular septum:21, TGA with ventricular septal defect (VSD):13, Taussig-Bing anomary (TBA):7, and double outlet right ventricle with noncommitted VSD:1]. Hospital death occurred in 1 patient (2.4% mortality rate) due to low cardiac output syndrome (LOS) caused by prolonged aortic clamp, who had TGA with VSD, coactation of aortic arch and right ventricular outflow obstruction. Four patients required re-operation (freedom from re-operation rate was 84.7%). Two had pulmonary stenosis and 1 needed right ventricular outflow tract (RVOT) reconstruction with transannular patch. His pulmonary valve used for previous VSD closure was diminished. The other required muscle resection of RVOT. Five patients presented grade II or more neo-aortic insufficiency and 2 of them were TBA. Both these TBA, rerouting of left ventricular outflow tract (LVOT) to the aorta was challenging, they showed LVOT obstruction postoperatively. One underwent aortic valve plasty and the other had residual VSD closure. None had coronary event or abnormality in coronary arteriography. Selection of surgical procedure should be based on morphologic features in order to improve surgical outcome. PMID- 18411690 TI - [Results of arterial switch operation for transposition of great arteries with regard to coronary pattern]. AB - One of the factors determing the postoperative results of arterial switch operation (ASO) for complete transposition of great arteries (TGA) is coronary event (CE). We analyzed the risk factors of CE after ASO in patients whose coronary patterns excluding Shaher type 1. The subjects were 44 patients operated from 1994 to 2007. As for the coronary reconstructive technique, procedures by Mee or Aubert were conducted on patients with Shaher 5A intramural type. The conventional technique was conducted on other patients, with or without trapdoor technique. In all patients, the pulmonary artery was reconstructed mainly with autopericardium using Lecompt maneuver. The mean observation period was 4.0 +/- 3.1 years, and the number of patients showing CE was 14 (32%) at 1.6 years after surgery on the average. The risk factors of CE and the operative procedures were discussed. PMID- 18411691 TI - [Surgical treatment of transposition of great arteries: the coronary buttons transfer using medially based trapdoor flaps]. AB - The arterial switch operation (ASO) has become the primary surgical approach used for correction of transposition of the great arteries. All the prerequisites for a successful ASO were recognized in time and dealt with, which allowed general acceptation of the technique. We report on our technique for the procedure and the result to date. From January 1991 to January 2008, a total of 100 patients underwent ASO at our unit using medially-based trapdoor flap method. The neo pulmonary artery (PA) was reconstructed using a single rectangular pericardial patch. The initial patient having intramural coronary artery died due to ischemic event after Aubert procedure. Three patients had re-right ventricular out flow tract repair (RVOTR) in a long-term follow-up period. There was no significant aortic insufficiency, no ischemic event and no lethal arrhythmia. PMID- 18411692 TI - [Novel coronary transfer technique for transposition of the great arteries with Planche type II coronary anatomy]. AB - The arterial switch operation has evolved to become the treatment of choice for most of transposition of the great arteries and some types of double outlet right ventricle. Technical improvement in the coronary transfer is the major contribution to the evolution of this procedure. We proposed a novel technique of coronary transfer for the patients with rare but difficult coronary anatomy of Planche type II. We believe it could become one of the options of coronary transfer technique and contribute to the completion of arterial switch operation with uncommon coronary artery patterns. PMID- 18411693 TI - [Technical modification and late results of arterial switch operation]. AB - The success of arterial switch operations for transposition of the great arteries largely depends on faultless coronary translocation and subsequent sufficient myocardial perfusion. Occurrence of late coronary obstructions after arterial switch operation was often reported. We developed the bay window technique as a useful adjunct to prevent coronary complications. Late results of this technique are described. No coronary events occurred (i.e. inclusive of coronary stenosis, myocardial infarction, and coronary death). The bay window technique is an innovative and simple surgical adjunct for translocating complex coronary arteries. PMID- 18411694 TI - [Long term outcome of arterial switch surgery for transposition of the great arteries: evaluation of the reconstruction of the pulmonary artery]. AB - We assessed the effect of reconstructing the pulmonary artery during arterial switch surgery for transposition of the great arteries on late pulmonary stenosis. Sixty-five patients who underwent Lecompte procedure between September 1991 and December 2006 were divided, by the procedure used chronologically to reconstruct the pulmonary artery, into group XP (single pantaloon patch with equine pericardium, n = 11), group P (direct reconstruction, n = 47), and group AP (single pantaloon patch with fresh autopericardium, n = 7). Outcome and pulmonary stenosis on the most recent ultrasound cardiography (UCG) were compared in the 3 groups. The median follow-up was 13, 7.5, and 1.3 years, respectively. Both early and late mortalities were 1.5% (1/65). Although percutaneous trans pulmonary angioplasty was necessary in 1, 13, and 3 patients, there was 1, 1, and 0 reoperation for pulmonary stenosis in the 3 groups, respectively. Pulmonary stenosis (pulmonary arterial maximum flow velocity > 3 m/sec on UCG) was present in 4 (40%). 14 (30%). and 3 patients (43%). Although there was no significant difference among the 3 procedures in preventing pulmonary stenosis 10 years after arterial switch surgery, direct reconstruction of the pulmonary artery may show a superior outcome, in particular, over 10 years after arterial switch surgery. PMID- 18411695 TI - [Long-term result of arterial switch operation for corrected transposition of the great arteries or double inlet left ventricle]. AB - The purpose of this study is to analyze the operative maneuver and long term outcome of the arterial switch operation (ASO) for congenitally corrected transposition of the great arteries (c-TGA) or double inlet left ventricle (DILV). Since October 1977, 221 patients had undergone ASO in National Cardiovascular Center, Japan. Of these, 8 patients underwent ASO as a part of double switch operation (DSO) for c-TGA, and 1 patient underwent ASO and ventricular septation for the DILV with a rudimentary right ventricle simultaneously. We retrospectively reviewed these 9 patients. Six patients had a past history of the pulmonary artery banding. Age at the time of ASO ranged from 6 months to 5 years (median 3 year). As a reconstruction of the pulmonary artery at the time of ASO, Lecompte maneuver was performed in 7 patients, and original Jatene procedure was performed in 2. Coronary transfer was done as usual in all patients. There was no early death, and 1 patient died 1 year after the operation due to chronic heart failure. Late complication related to the ASO was pulmonary artery stenosis (1 patient after DSO) and aortic regurgitation (1 patient after ventricular septation). PMID- 18411696 TI - [Minimally invasive cardiac surgery for transposition of the great arteries in neonates]. AB - We evaluated our results of arterial switch operation (Jatene) for complete transposition of the great arteries between May 2003 and October 2005 particularly concerning various operation-related durations. Twenty neonates were studied. The mean age and body weight were 11.6 +/- 2.7 (range 5 to approximately 15) days and 3.0 +/- 0.4 kg, respectively. Duration of anesthesia, operation, extracorporeal circulation (ECC), and aortic cross-clamp were 199.4 +/- 30.1, 162.7 +/- 29.9, 91.6 +/- 8.8 and 59.8 +/- 8.1 minutes, respectively. Time differences between anesthesia and operation, operation and ECC, ECC and aortic cross-clamp were calculated, and their correlations with the duration of anesthesia were investigated. The items, whose coefficient of correlation with anesthetic time was greater than 0.6 were operation time, ECC time, aortic cross clamp time, operation time minus ECC time, and operation time after ECC come-off. Furthermore, the operation time after ECC come-off was strongly correlated with plasma lactate concentrations and intraoperative bleeding. In conclusion, the time required for hemostasis and closure of the chest should be as short as possible. Therefore secure anastomoses with least hemorrhage possible is important. PMID- 18411697 TI - [Safety management of bronchoscopic examination and treatment]. AB - Safety management of bronchoscopic examination and treatment is devided into 3 phases. Patient information should be provided before the procedure. Also written informed conscent should be obtained after the information of advantage and disadvantage of examination/treatment. Bronchoscopists and well-trained nurses should manage both the procedure itself and patient condition. Through check up of patient is necessary to avoid late complications after the procedure. Due to the increase in the number of interventional bronchoscopies, the management of patients is getting more important. Safety management should be developed by wide range of medical team including bronchoscopists, nurses and technical staff. PMID- 18411698 TI - [The relation between the frequency of exercise oxygen desaturation following major lung resection and the predictions of postoperative pulmonary function]. AB - The predictors of oxygen desaturation during exercise in patients submitted to major pulmonary resection for lung cancer are to be determined. We analyzed retrospectively the relation between the oxygen saturation by pulse oxymetry (Spo2) during exercise and the predictions of postoperative pulmonary function. A hundred twenty-two patients with lung cancer who underwent lung resection from January 1999 to May 2004 were included (79 men, 43 women, average age 66.9 +/- 9.2). A fall over 5% in Spo2 during exercise was termed 'desaturation'. Twenty eight patients developed desaturation [group D(+)] and 94 patients did not [group D(-)]. We compared the predictions of postoperative pulmonary function (%ppoVC, %ppoFEV1.0, %ppoDLco) between these 2 groups. As a result, only %ppoDLco was significantly different between 2 groups [D(+) 68.7 +/- 19.1%, D(-) 83.8 +/- 24.9%]. Patients with poor %ppoDLco are at increased risk to develop a postoperative exercise oxygen desaturation. PMID- 18411699 TI - [A prosthetic ring annuloplasty with edge-to-edge repair for a treatment of severe tricuspid regurgitation]. AB - A 59-year-old woman admitted to our hospital with shortness of breath and edema of the lower extremities was diagnosed with right ventricular failure stemming from severe tricuspid valve regurgitation (TR). She had undergone mitral valve replacement (MVR) with a mechanical valve at the age of 42. The approach to the heart was established via a right thoracotomy at the 4th intercostals space. A beating heart cardiopulmonary bypass procedure was performed in which tricuspid valve repair was performed with the edge-to-edge repair and MC3 annuloplasty system. The operative course was uneventful. This technique may be feasible and clinically effective in the treatment of severe TR. PMID- 18411700 TI - [Uncontrollable midbrain hemorrhage due to brain metastasis of pulmonary pleomorphic carcinoma]. AB - A 54-year-old man who had been treated hypertension admitted our hospital due to midbrain hemorrhage. Pulmonary abnormal shadow was also found by chest radiological examinations and it was diagnosed as stage IB (T2N0M0) non-small cell lung cancer. Right upper lobectomy with combined resection of azygos vein and parietal pleura facing to the tumor was conducted because the direct invasion of the tumor was strongly suspected. Pathological diagnosis was pleomorphic carcinoma. Although, postoperative course was uneventful, midbrain hemorrhage recurred 3rd postoperative day. Since the hemorrhage continued in spite of the conservative treatment, the craniotomy was performed on the 13th postoperative day. The pathological examination revealed the hemorrhage to be caused by the brain metastasis. PMID- 18411701 TI - [Hemoptysis one year after video-assisted thoracoscopic bullectomy for spontaneous pneumothorax: report of a case]. AB - A 63-year-old man who had underwent video-assisted thoracoscopic bullectomy for left spontaneous pneumothorax 1 year before developed recurrent hemoptysis. Chest computed tomography showed previous stapling of the subsegmental bronchus in the left apico-posterior segment Bronchial arteriography showed hypervascularization of bronchial artery in the left upper segment and pooling of contrast medium along the staple-suture line. Video-assisted thoracoscopic apico-posterior segmentectomy was performed successfully. Pathological examination revieled hemosiderin lining along the surgical stump of B(1+2)cii, neither with infection nor infarction. These findings suggest that mechanical stapling of B(1+2)cii induced ischemia in the peripheral lung parenchyma causing bronchial hypervascularization. Late onset hemoptysis should be kept in mind as a complication after bullectomy with a mechanical stapler. PMID- 18411702 TI - [Regulation by tissue plasminogen activator of rewarding effects of drugs of abuse]. AB - Drugs of abuse acutely modulate the activity of mesolimbic dopaminergic neurons, projecting from the ventral tegmental area of the midbrain to the nucleus accumbens (NAc). Tissue plasminogen activator (tPA) is a serine protease that catalyzes the conversion of plasminogen (plg) to plasmin. Here we show that this protease system participates in the rewarding effects of morphine, methamphetamine (METH) and nicotine. A single morphine treatment induces tPA mRNA and protein expression in the NAc. Morphine-induced conditioned place preference and hyperlocomotion are significantly reduced in tPA-deficient (tPA-/-) and plg deficient (plg-/-) mice, being accompanied by a loss of morphine-induced dopamine release in the NAc. Repeated METH treatment also induces tPA mRNA expression in the NAc. METH-induced conditioned place preference and behavioral sensitization after repeated METH treatment are significantly reduced in tPA-/- mice compared with those in wild-type mice. Finally, we show that the tPA-plasmin system regulates nicotine-induced reward and dopamine release by activating protease activated receptor-1 (PAR1). Nicotine-induced dopamine release is markedly diminished in tPA-/- mice. Furthermore, plasmin activates PAR1 and nicotine induced conditioned place preference and dopamine release are diminished in PAR1 deficient mice. Our findings suggest that targeting the tPA-plasmin-PAR1 system would provide new therapeutic approaches to the treatment of drug dependence. PMID- 18411703 TI - [Neurochemical mechanisms for development of psychological dependence on volatile organic solvents]. AB - Abuse of volatile organic solvents among youth remains a major social problem. Organic solvents are cheap and relatively easy to obtain, so they carry the risk of becoming a "gateway drug" for users. The effect of repeated inhalation of toluene on subsequent responses to other drugs of abuse is unclear. In the present study, we investigated the effect of toluene inhalation on methamphetamine-induced behavioral change using a newly developed sealed inhalation shuttlebox. The influence of the cyclic AMP response element binding (CREB) protein expression following toluene inhalation was also examined. Mice were exposed to toluene or air once daily for five days. Methamphetamine produced significant hyperlocomotion in air-exposed mice. This stimulatory effect of methamphetamine was significantly enhanced following repeated inhalation of toluene. Furthermore, repeated toluene inhalation increased the levels of CREB proteins in the limbic forebrain. The present study demonstrated that adaptation of the adenylate cyclase system following repeated toluene inhalation might be involved in the expression of behavioral sensitization to subsequent methamphetamine administration. Inhalant abuse could thus be associated with the risk of other substances of abuse. PMID- 18411704 TI - [Potential genetic predictors for individual vulnerability to substance dependence]. AB - Development of substance dependence is influenced by multiple factors, e.g. pharmacological effects on mental status, environmental and individual factors. Among them genetic factors were found to have greater affect on substance dependence than expected which inheritance rate was calculated as 0.7 or more. However, more precise information on genetic mechanisms underlying substance dependence is still unknown. In Japan, methamphetamine has been the most popular illicit drug. The JGIDA study for genetic factors of methamphetamine dependence/psychosis started in 2001, and revealed many potential genetic predictors for individual vulnerability to methamphetamine dependence and variation of clinical phenotypes. As to susceptibility to drug dependence, 17 genetic variants were identified. A certain SNP or haplotype of the DTNBP1, GSTM1, GSTP1, glycine transporter-1 gene produce risks and that of serotonin transporter, AKT1 and CYP2D6 gene produce negative risks. As to risks of rapid onset of methamphetamine psychosis, worse prognosis or complication of spontaneous relapse, the dopamine D2 receptors, dopamine transporter, monoamine oxidase-A, catechol-O-methyltransferese, SOD2, NQO2, PICK1 gene were identified. Odds ratios of these positive or negative risks ranged from about 0.2 to 15. These findings must be informative for drug dependence rehabilitation programs and psycho-education of substance dependence. PMID- 18411705 TI - [Minocycline as a therapeutic drug for methamphetamine use disorders]. AB - Use of methamphetamine and 3,4-methylenedioxymethamphetamine (MDMA) is an extremely serious and growing problem throughout the world, including Japan. Antipsychotic drugs have been used for psychotic symptoms associated with these abused drugs. However, there are currently no particular pharmacological treatments for the wide range of symptoms associated with these abused drugs. Recently, we reported that the second generation antibiotic drug minocycline can attenuate behavioral abnormalities and neurotoxicity in the brain after administration of methamphetamine or MDMA. In this review, we discuss minocycline as a new potential therapeutic drug for schizophrenia as well as psychosis associated with these abused drugs. PMID- 18411706 TI - [H magnetic resonance spectroscopy investigation of the anterior cingulate cortex and prefrontal cortex in chronic pain patients]. AB - We investigated the absolute concentration of N-acetylaspartate (NAA) in the anterior cingulated cortex (ACC), and prefrontal cortex (PFC) in chronic pain patients by proton magnetic resonance spectroscopy (1H-MRS). 1H-MRS was performed with a 1.5T MR system on a voxel in the thalamus, ACC, and PFC bilaterally, in 48 chronic pain patients and 23 normal control subjects. We measured the absolute concentration of NAA using an LC-Model. There were 12 patients whose NAA concentration in the ACC or PFC was significantly, lower than the mean NAA concentration of the normal control subjects. Ten of these 12 patients needed a psychological approach. 1H-MRS suggests that NAA concentration in the ACC or PFC is associated with the necessity of a psychological approach. 1H-MRS may serve as a useful non-invasive tool for evaluating neural activity of the thalamus, anterior cingulated cortex, and prefrontal cortex in chronic pain patients. PMID- 18411707 TI - [Descending facilitation in chronic stress and chronic pain state]. AB - The spino-thalamic tract consists of two systems; the lateral system terminates in the somato-sensory cortex, and participates in the sensory discrimination of pain, and the medial system terminates in the anterior cingulated cortex (ACC) and insular cortex (IC) to mediate affective components of pain. Persistent pain induces plastic changes in cortical neurons, especially in the ACC and IC. Activation of these neurons is transmitted to the periaqueductal gray and rostroventromedial medulla (RVM) (descending pain control system). This system has long been considered to exert descending inhibition, but recent studies revealed that it also causes facilitation in certain pathological conditions. A variety of stressful stimuli have been shown to affect pain sensitivity. We demonstrated that chronic restraint stress induced thermal hyperalgesia in rats, in which phosphorylated ERK and levels of tryptophan hydroxylase, a key enzyme of 5-HT production, were increased in the RVM. 5HT released from the bulbospinal neurons may exert facilitatory effects on spinal nociceptive processing probably through 5HT3 receptors. Patients suffering chronic pain originating from deep tissues, such as temporo-mandibular disorder, fibromyalgia, or low back pain, often complain of pain and tenderness in various parts of the body. We injected complete Freund's adjuvant into a temporo-mandibular joint of rats unilaterally, and then injected 5% formalin into the ipsilateral or contralateral masseter muscle 2 weeks later. Pain-related behavior and neuronal activation in the spinal trigeminal nucleus were enhanced on both sides compared to those in non inflammatory controls. Systemic enhancement of pain and hyperalgesia induced by unilateral joint inflammation may have been caused by the central sensitization and descending facilitation. PMID- 18411708 TI - [Neuronal basis for pain-induced aversion]. AB - Pain is a complex experience composed of sensory and emotional components. We demonstrated the differential patterns of c-fos mRNA induction by chemical somatic (formalin) and visceral (acetic acid) noxious stimuli in the rat amygdaloid nuclei, the brain regions implicated in emotion. We also showed that conditioned place aversion (CPA) induced by formalin was abolished by the lesion of the basolateral (BLA) or central (CeA) amygdaloid nucleus, while the acetic acid-induced CPA was abolished by the CeA-, but not BLA-, lesion. These results suggest the differential contribution of the BLA and CeA to the negative emotional component of chemical somatic and visceral pain. We demonstrated the critical involvement of intra-BLA glutamatergic transmission via NMDA receptors in formalin-induced aversion. Intra-BLA morphine suppressed this glutamatergic transmission as well as somatic pain-induced aversion. Furthermore, we examined the role of the bed nucleus of the stria terminalis (BST), one of the brain regions forming the extended amygdala. Excitotoxic lesion of the BST as well as intra-BST injection of a beta-adrenoceptor antagonist timolol suppressed the formalin-induced CPA, suggesting that noradrenergic transmission via beta adrenoceptors within the BST plays an important role in the emotional component of pain. PMID- 18411709 TI - [Individual differences in analgesic effects of narcotics]. AB - Narcotic analgesics have been widely used for management of severe pain, especially for cancer pain. Most of these drugs are opioids, and they show their analgesic effects by acting through opioid receptors. Significant individual differences in opioid sensitivity can hamper effective pain treatments and increase side effects, which is associated with decreased quality of life. It is thought that genetic factors may affect individual differences in opioid sensitivity. Recent studies using various inbred and knockout mice have revealed that the mu-opioid receptor (MOP) plays a mandatory role in the analgesic properties of opioids. There is also increasing evidence that differences in the sequence of the MOP gene might significantly affect the amount of MOP gene mRNA expression and sensitivity to opioids. Furthermore, it can be thought that individual differences in opioid sensitivity are caused by genetic differences in not only MOP but other biomolecules, such as endogenous opioid peptides, molecules related with metabolic process and second messenger systems. Rapid advances in this research field are leading to a better understanding of relationships between gene polymorphisms and opioid sensitivities, which, in turn, will allow us to more accurately predict opioid sensitivity and opioid requirements in individual patients. PMID- 18411710 TI - [Infection and immunity of the eye]. PMID- 18411711 TI - [The cornea: stasis and dynamics]. AB - The physiological roles of the cornea are to conduct external light into the eye, focus it, together with the lens, onto the retina, and to provide rigidity to the entire eyeball. Good vision thus requires maintenance of the transparency and proper refractive shape of the cornea. Although the cornea appears to be a relatively static structure, dynamic processes operate within and around the cornea at the tissue, cell, and molecular level. In this article, I review the mechanisms responsible for maintenance of corneal homeostasis as well as the development of new modes of treatment for various corneal diseases. I. The static cornea: structure and physiological functions. The cornea is derived from ectoderm, so that it can be considered as transparent skin. It is devoid of blood vessels and manifests the highest sensitivity in the entire body. The surface of the cornea is covered by tear fluid, which serves both as a lubricant and as a conduit for regulatory molecules. The cornea is also supplied with oxygen and various nutrients by the aqueous humor and a loop vascular system in addition to tear fluid. The cornea interacts with its surrounding tissues directly as well as indirectly through tear fluid or aqueous humor, with such interactions playing an important role in the regulation of corneal structure and functions. The resident cells of the cornea-epithelial cells, fibroblasts (keratocytes), and endothelial cells--also engage in mutual interactions through network systems. These interactions as well as those with infiltrated cells and regulation by nerves contribute to the maintenance of the normal structure and functions of the cornea as well as to the repair of corneal injuries. II. The dynamic cornea: maintenance of structure and functions by network systems. Developments in laser and computer technology have allowed observation of the cells and collagen fibers within the cornea. Furthermore, progress in cell and molecular biology has allowed characterization of dynamic network systems-including cell-cell and cell extracellular matrix interactions as well as cytokines and neural factors-that contribute to the maintenance of corneal transparency and shape. III. Disruption of network systems: persistent corneal epithelial defects and corneal ulcer. Selection of the appropriate treatment for pathologic lesions of the cornea and the accompanying decrease in visual acuity requires localization of the lesion with regard to the epithelium, stroma, or endothelium of the cornea. In certain instances, however, it is not possible to determine the cause of the problem within the cornea. In such cases, the cause of the pathologic lesion and the target for treatment may lie in the surrounding tissues or environment. For example, corneal epithelial wound healing may be delayed, leading to the development of persistent epithelial defects, as a result of disruption of intercellular junctions between epithelial cells, an abnormality of the corneal basement membrane, altered concentrations of various cytokines in tear fluid, a lowered corneal sensation, or allergic reactions in the lid conjunctiva. Loss of corneal epithelial barrier function can further allow inflammatory cytokines present in tear fluid, together with infiltrated cells, to activate keratocytes and elicit excessive degradation of collagen in the stroma, thereby giving rise to corneal ulcer. IV. Development of new drugs for corneal diseases. We have attempted to apply the results of basic scientific research to the development of new drugs for corneal diseases that remain difficult to treat. The process of authorization for new drugs from the Ministry of Health, Labor, and Welfare takes more than two decades, however. The path from the bench to clinical practice is thus a long one. 1. Development of eyedrops for treatment of persistent corneal epithelial defects. We demonstrated the clinical efficacy of fibronectin eyedrops for the treatment of persistent epithelial defects of the cornea. However, the possibility of blood-borne infections has interfered with the development of serum-derived fibronectin as a drug. An automated machine for the preparation of autologous fibronectin eyedrops has therefore recently been developed. Furthermore, in seeking an alternative to fibronectin eyedrops, we are investigating the effects of a peptide corresponding to the second cell-binding domain of fibronectin on corneal epithelial wound healing. Considering that urokinase-type plasminogen activator may be expressed at the site of corneal epithelial defects and facilitates epithelial migration, the potential clinical application of annexin V, which stimulates the secretion of urokinase-type plasminogen activator for the treatment of persistent corneal epithelial defects is also now under investigation in Japan. 2. Development of eyedrops for treatment of neurotrophic keratopathy. Substance P, a neurotransmitter, stimulates corneal epithelial migration in a synergistic manner with insulin-like growth factor (IGF)--1. We have shown that eyedrops containing both the substance P-derived peptide FGLM-amide and the IGF-1--derived peptide SSSR are effective for the treatment of persistent corneal epithelial defects in individuals with diabetic keratopathy or neurotrophic keratopathy, both of which are associated with a reduction in corneal sensation. 3. Development of drugs for corneal ulcer. Treatment of corneal infection with antibiotics does not necessarily halt the process of corneal ulceration, which is characterized by excessive degradation of stromal collagen, or resolve persistent corneal epithelial defects. In addition to eyedrops for the treatment of persistent corneal epithelial defects, we have therefore also been working on the development of new drugs for the treatment of corneal ulcer. To this end, we have established an experimental system in which corneal fibroblasts are cultured in a three-dimensional collagen gel. With this system, we have shown that triptolide and steroids inhibit collagen degradation by corneal fibroblasts. Triptolide or its derivatives are thus potential drugs for the treatment of corneal ulcer and would work by acting directly on corneal fibroblasts rather than by inhibiting the secreted enzymes(matrix metalloproteinases) responsible for collagen degradation. PMID- 18411712 TI - [Studies on clinical pathophysiology of pseudophakic/aphakic eyes--a journey of 4 decades]. AB - My prime years as an ophthalmologist began as intraocular lenses (IOLs) were just entering into the developmental stage, and I took on as my mission to contribute to perfecting safe and reproducible cataract/IOL implantation surgery. Identifying surgical and/or IOL-related complications consumed time; however, these complications soon became predictable and even preventable with the use of sensitive biological parameters and preclinical evaluation. This was a simple goal for me to pursue my studies on cataract/IOL implantation surgery. I discuss in this review article, based on my previous research, clinico-pathophysiological problems of these intra- and postoperative eyes. The early phase of cataract/IOL implantation surgery development began with a debate as to which is physiologically superior: intracapsular cataract extraction (ICCE) or extracapsular cataract extraction (ECCE). From the perspective of transporting substances from intraocular fluids to extraocular space, which we studied using a nonphysiological substance, fluorescein, ECCE was confirmed to be physiologically superior to ICCE. The transport mechanism of both physiological and nonphysiological substances from intraocular fluids (such as vitreous and aqueous humor) is believed to be related to the pathogenesis of various ocular disorders. Following the fluorescein study, I next focused my attention on biosynthesis and active transport of prostaglandin (PG), which are inflammatory mediators. My studies revealed that PG were more likely to accumulate in ICCE eyes than in ECCE eyes; higher intraocular concentration of PG was also confirmed in eyes with persistent aphakic or pseudophakic cystoid macular edema (CME). While conducting the above studies and having made some observations, I postulated another hypothesis on the pathogenesis of aphakic or pseudophakic CME as follows: topical application of nonsteroidal antiinflammatory drugs (NSAIDs) to eyes with PG, which are biosynthesized intra- and postoperatively during the healing process of uveal tissues and lens epithelial cells, prevents CME. Based on this hypothesis experimental studies were then started, and in 1977 I became the first in the world to prove that topical application of indomethacin, one of the NSAIDs, controls the incidence of CME in ICCE eyes. Thereafter, some 40 follow-up studies have been conducted worldwide, and recent meta-analysis has established the efficacy of indomethacin. Macular edema and CME are recently of significant interest as complications in various ocular disorders. Compared to other forms of CME, the pathophysiology of CME associated with aphakic/ pseudophakic eyes is relatively simple, its natural history is well understood and its reproducibility is high. It is possible that the other forms of macular edema or CME having more complicated pathogenesis may be interpreted by understanding the formation mechanism of aphakia/pseudophakic CME. Our studies have shown how chemical mediators (PG) are systematically involved in the development of aphakic/pseudophakic CME, and that they concurrently cause blood-aqueous barrier disruption and CME, decrease oscillatory potential of the full field ERG, and decrease choroidal blood flow at an early postoperative period, and this has recently been proven. All these phenomena, however, can be effectively prevented by topical application of NSAIDs. I believe these findings provide significant information when considering the pathogenesis and treatment of CME associated with other ocular disorders. Using the primitive method of an early phase, I discovered that anti-PG eye drops can treat disrupted blood-aqueous barrier, and confirmed that the blood-aqueous barrier function is indeed a very sensitive function. I next applied fluorophotometry and laser flaremetry. Using blood aqueous barrier function as a parameter, the following were evaluated: consensual reaction of blood-aqueous barrier disruption, method of IOL fixation, racial differences in disruption of the aqueous barrier function, drugs used perioperatively, biocompatibility of IOL materials, and effects of preservative agents. Research on preservative agents disclosed that the preservative agent in anti-glaucoma drops more strong by induced pseudophakic CME than the anti glaucoma agent itself. Thus, this introduced a new concept called Our desire to closely observe the endosurface of the iris, ciliary processes and anterior vitreous face, all of which are closely related to phacoemulsification techniques, posterior chamber lens fixation, and active transport of PG, led me to the development of "Posterior video technique" (Miyake-Apple View). The technique since then has been used to evaluate cataract surgical techniques, to analyze complications, to review IOL designs and fixation techniques, to pre clinically evaluate surgical devices, and to study variations of local anatomy related to cataract/IOL surgery. The method is also useful as an educational as well as a presentational tool, and it has now been accepted world-wide. The pathogenesis of aphakic/pseudophakic CME, physiological evaluation centering on blood-aqueous barrier function, and preclinical evaluation using the Posterior video technique have all played a significant role in establishing today's safe cataract/IOL implantation surgery. PMID- 18411713 TI - [Herpes simplex virus latency, reactivation, and a new antiviral therapy for herpetic keratitis]. AB - Although many factors that trigger the herpes simplex virus (HSV) reactivation from latency have been reported, how HSV resides in a latent state in the normal human cornea still needs to be defined. We therefore conducted a series of studies regarding various aspects of HSV infections. To understand how patients subjectively perceived changes in their daily life that could have induced HSV reactivation, we first performed a comprehensive survey on the subjective factors in patients who had experienced recurrent herpetic keratitis. The result of our survey revealed that stress, lack of sleep, shoulder stiffness, and physical fatigue were the key factors. There were various causes for stress, and stress associated with reactivation often occurred between spring and summer. Regarding HSV latency in the normal cornea, we used real-time polymerase chain reaction (PCR) to determine the presence of HSV in the donor and host corneas. The findings showed that on average, those host corneas with a history of HSV keratitis had 1.6 x 10(4) copies/mg of HSV DNA, while the host corneas without a history and the donor corneas had 8.7 and 4.9 x 10(2) copies/mg of HSV DNA, respectively. Based on these observations, it is reasonable to infer that latent viruses could have resided in a normal cornea without a history and were transmitted to a host cornea through corneal transplantation. We also quantified the virus load in tears before and after ocular surgery (one week after corneal transplantation or the next day after vitreous surgery). Our results indicated that both the detection rate and the average copy number of HSV DNA had a tendency to increase postperatively. Moreover, we tried to differentiate the HSV strains that were involved in the recurrent lesions. In only one of the studied cases, could we find a single different nucleotide between two HSV strains. It seemed possible that two different strains of HSV had set off the same episode of reactivation. In recent years, chemokines have become known for their action in mediating inflammatory diseases. We suspected that chemokines might also play a role in the antiviral mechanism and examined the chemokine-derived antiviral activity. We used eight chemokines, including RANTES/CCL5, MIP-lalpha/ CCL3, and MIP-1beta/CCL4, in a murine HSK model with Vero cells. These chemokines directly bound to HSV and the chemokine-bound HSV was later resisted by the neutralizing antibody of envelope protein gB. Furthermore, by electron microscope analysis, it became clear that these chemokines had cut an opening in the HSV envelope. Consequently, these chemokines had significantly inhibited the HSV infection on Vero cells. In addition, the virus load in tears was decreased and the corneal opacity was less severe. We concluded in that study that during early infection, chemokines accumulated in the corneal stroma have the ability to protect cells and tissues from HSV infection. As for antiviral therapy, acyclovir (ACV) eye ointment has been effective for patients with herpetic keratitis. However, patients often find it difficult to successfully follow the treatment due to the required frequent application and the blurred vision after application. On the other hand, valaciclovir (VCV), which is the oral prodrug of ACV, has become commercially available in recent years for treating nonocular herpetic diseases. We therefore examined and compared the efficacies of oral VCV, oral ACV, ACV eye ointment, and ACV eye drops in a murine keratitis model; the group treated with oral VCV did show a significantly good antiviral effect. We have proved that oral VCV can be a beneficial alternative antiviral therapy for patients with difficulty in complying with the ACV eye ointment treatment. PMID- 18411714 TI - [Treatment strategy for rejection-free corneal transplantation--transition from full-thickness corneal transplantation to corneal endothelium transplantation]. AB - The avoidance of allograft rejection is the most critical factor for favorable surgical outcome after corneal transplantation. We report experimental data including distribution of white blood cells in human corneas for rejection-free corneal transplantation. We focused on leukocyte trafficking based on the immunological mechanism leading to allograft rejection in a mouse full-thickness corneal transplantation model. We identified two chemokine-receptors, CCR1 and CCR7 which are functionally relevant to the occurrence of allograft rejection. These chemokine receptors can be new targets for the suppression of allograft rejection after full-thickness corneal transplantation. In the human corneas, bone marrow-derived dendritic cells and monocyte-lineage cells reside constitutively in the normal epithelium and stroma, and may be associated with direct recognition of allo-antigen after corneal transplantation. We established a mouse model in which cultured allocorneal endothelium was transplanted onto a bullous keratopathy recipient cornea. During the follow-up period, the transplanted cultured allo-corneal endothelium did not show any sign of allograft rejection. Our findings demonstrated that a rejection-free mechanism is due not to suppression of immunity or to lack of response, but to failure to recognize the existence of resistance. Realization of the clinical application of cultured allo-corneal endothelium transplantation may be a shortcut to ideal rejection free corneal transplantation. PMID- 18411715 TI - [Association of ocular inflammation and innate immune response]. AB - Immune response has been divided into innate immunity and acquired immunity. We focused on the role of innate immunity during the formation of uveitis and choroidal neovascularization (CNV)-related diseases. To carry out a comprehensive analysis of ocular inflammatory responses in patients with uveitis, vitreous fluid was analyzed using a microbead-based multiplex ELIZA system. We found that cytokines which were related with innate immunity were elevated, but cytokines which were related with acquired immunity were not. We also found that the role of IL-17 was to produce Th17 cells in the chronic phase of experimental uveitis. Next, we investigated the role of the natural killer (NK) T cells which restrict CD1 and participate in the innate immune response in laser-induced experimental CNV. We studied the CNV formation in two independent NK T cell-deficient strains of mice, CD1 knockout (KO) mice and Jalpha18 KO mice, and found that both KO mice showed significant reduction of the effects of experimental CNV. After laser treatment, both CD1 KO mice and Jalpha18 KO mice showed a decrease in the expression of vascular endothelial growth factor (VEGF) expression in retina and choroid. Interestingly, intravitreous inoculation of a galactosylceramide (alphaGalCer), which is the ligand of NK Tcells, inhibited CNV in C57BL6 mice. Collectively, we conclude that NK T cells play an important role in forming CNV as one of the inducers of VEGS. Because NK T cells bear the potential to regulate immune response, alphaGalCer might activate NK T cells differently to produce angiostatic factors and have a therapeutic potential in vivo. During the clinical process of CNV-related diseases, not only CNV formation, but also subretinal scarring is thought to be another important step. We thus established the experimental model of subretinal scaring by injecting peritoneal exudating macrophases into the subretinal space. This scaring was inhibited by inoculation of anti-IL-6 antibody and micro bubbles into the vitreous cavity following low power ultrasound treatment through the cornea. PMID- 18411716 TI - [Gastrointestinal protective barrier]. AB - Current views on the protective function of the gastrointestinal barrier are based on the capacity of its surface and glandular epitheliocytes to synthesize the mucins. The most significant types of gel-forming mucins are MUC 5AC, MUC 5B, MUC 6 and MUC 2. The type of the mucin is cell-specific and is determined by the character of cell phenotype. The change of the mucin type may be the major factor in the development of pathology. The protective mucin function is realized with the participation of trefoil peptides. The problem of the mechanism of mucin trefoil peptide macromolecular complex formation in normal and pathological states is discussed. PMID- 18411717 TI - [Cytoarchitectonics, neuronal organization and the influence of gender factor on neuronal dendroarchitectonics of the amygdala posterior medial nucleus]. AB - The aim of this study was the analysis of cytoarchitectonics, neuronal organization and gender factor influence on neuronal dendroarchitectonics of amygdala posterior medial nucleus. The study was performed on 50 adult Wistar rats (25 male and 25 female) using Nissl and Golgi staining methods. The nucleus is shown to be formed by large neurons, which in silver-impregnated sections, had the characteristics of long-axon sparsely- and densely-branched neurons. In male rats, dendrites of long-axon sparsely-branched neurons had greater branching indexes, while those of long-axon densely-branched neurons were longer than in females. PMID- 18411718 TI - [Structural and metabolic changes in the rat hypothalamic histaminergic neurons induced by the bile loss]. AB - The aim of the study was the estimation of structural and metabolic changes in histaminergic neurons of rat hypothalamic E2 nucleus induced by total external bile drainage. The investigation was carried out on male Wistar rats (n=45). The control group comprised the sham-operated animals, in which the physiological bile drainage was preserved during the whole experimental period. Quantitative histological and histochemical methods were used. In serial frontal cryostat sections of posterior hypothalamus, the activity of the following enzymes was demonstrated histochemically: monoamine oxidase B, succinate dehydrogenase, NADH- dehydrogenase, NADPH-dehydrogenase, glucose-6-phosphate dehydrogenase, lactate dehydrogenase and acid phosphatase. Morphometric study of histaminergic neurons was performed in thionin-stained sections. It was found that total external bile drainage resulted in a temporary reduction of the sizes and rounding of neuronal perikarya. Metabolic changes were detected already after 1 day of bile loss, and they were found to progress henceforth. All the pathways of energy metabolism were suppressed, while the acid phosphatase activity was increased, on day 5. PMID- 18411719 TI - [Intracerebral progression of the transplanted rat C6 glioblastoma cells pretreated with neuropeptides and MAPK inhibitor]. AB - The authors have monitored C6 glioma cell invasive growth, proliferation and transcriptional regulation after pretreatment with endothelin-1 and ERK1/2 specific inhibitor PD98059. To explore proliferation of C6 glioma cells in different growth conditions, they were treated in vitro with endothelin-1 and implanted into the brain. In vitro studies have indicated that PD98059 inhibited the proliferation of cultured C6 glioma cells and induced the activation of E2F1 and Myc-Max transcriptional factors. Endothelin-1 strongly increased C6 glioma cell proliferation. The model used in this study is experimental, but it may provide an insight into the specific behavior of in vitro cultured invasive cells. PMID- 18411720 TI - [Changes of the ventral dendrite of a goldfish Mauthner neurons induced by optokinetic stimulation]. AB - It was recently demonstrated that following the optomotor drum, rotating contralaterally to the turn side, spontaneously preferred by the goldfish fry, inverted their motor asymmetry. The study of Mauthner neuron (MN) structure using the histological method of 3D reconstruction, and the measurements of right and left MN in these fishes, has detected the 3-fold decrease of the volume of the ventral dendrite (VD) of ipsilateral MN, which was reciprocally related to MN functional activity. At the same time, total volume of contralateral MN remained larger than that of ipsilateral one by one quarter, which was the case in intact fishes. It is suggested that the decrease in the size of VD, which receives the afferentation from the contralateral eye, that follows the moving stripes of the optomotor drum, is the result of a specific contralateral visual stimulation and is the first evidence of the possibility of MN natural stimulation through VD. PMID- 18411721 TI - [Parasympathetic innervation of proximal parts of the colon in cat]. AB - The localization and morphometric features of efferent parasympathetic neurons of the vagus dorsal motor nucleus and of the spinal sacral parasympathetic nucleus innervating the area of ileocaecal sphincter, ascending and transverse colon, were investigated. In urethane anaesthetized cats, the solution of horseradish peroxidase was injected under the serosa of the indicated areas of colon. In 48 hours animals were transcardially perfused with a fixative solution. Sections of the medulla oblongata and the sacral spinal cord were stained using Mezulam's technique (1978). It was shown that all the areas of the colon studied received parasympathetic innervation from the neurons of the ventrolateral part of the vagus dorsal motor nucleus, which were uniform according to their morphometric characteristics. The number of neurons in this group, sending their axons to the ileocaecal area, was greater than the number of neurons, innervating ascending colon. Second group of neurons, that was represented by smaller cells, was located in the same part of the nucleus and innervated transverse colon. Transverse colon had an additional parasympathetic supply from the neurons of the spinal sacral parasympathetic nucleus. PMID- 18411722 TI - [Reaction of the rat brain capillaries to immobilization stress]. AB - Using calcium-adenosine triphosphate histoangiological method of Chilingarian, the morpho-functional state of the capillary limb of the microcirculatory bed in rat brain was studied at various time intervals after the exposure to experimental immobilization stress (fixation of the animals on the back for 2 hours). The analysis of morphometric data has show that in comparison with intact animals, immediately after the stress the capillaries were contracted by 17.2%, while on the second day their compensatory dilatation by 2.5% took place, which was later followed by a small contraction of 5.6%. Morphometric parameters of the placid and aggressive groups of animals indicate that the behavioral stereotype of placid animals secures more sparing physiological response to stress. It is suggested that the differences in capillary dysfunction significantly depend on the degree of lesions of the neuronal systems, which are involved in microcirculation control. PMID- 18411723 TI - [Structural changes of rat myocardium in the mother-fetus system exposed to cadmium]. AB - Myocardium structure and the pattern of cardiomyocyte cellular and intracellular changes were studied in rats following the exposure of the mother-fetus system to cadmium. Rats were intraperitoneally injected with cadmium sulfate during days 1 16 of pregnancy. Myocardium of the left ventricle was studied in pregnant females and fetuses on day 20 of gestation. Myocardium changes in cadmium-exposed animals included the reduction of cardiomyocyte and blood vessels relative volume in mother and fetus. The damage of the nuclear and nucleolar apparatus was detected, together with the signs of diffuse edema of cardiomyocyte myofibrils and dilatation of intercellular spaces within the myocardium. This changes in cadmium exposed animals are indicative of the development of regenerative-plastic myocardium insufficiency in the mother-fetus system. PMID- 18411724 TI - [Structural and functional changes of the loose connective tissue and macrophage system in experimental crush-syndrome]. AB - Cells and the extracellular matrix of the loose connective tissue as well as the macrophage system were studied in experimental crush-syndrome in 50 rats using histological, histochemical and electron microscopical methods. The results of the investigation have shown that the components of the loose connective tissue and the macrophage system actively responded to 1-hour-long hindlimb compression. Severe structural and functional injury of the cells and extracellular matrix of the subcutaneous loose connective tissue and of the stroma of different organs (lung, heart, kidney) was detected on dayl, was especially pronounced on day 7 and partly regressed by day 30 after the decompression. These changes included the structural damage of fibroblasts, changes in the number and degranulation of the mast cells, inhibition of RNA synthesis in plasmoblasts and lymphoblasts and the destruction of the collagen, elastic and reticular fibers. The number of macrophages in the liver, lungs, spleen, lymph nodes was reduced on days 1 and 7 after the decompression, in some organs this effect persisted until day 20. The activity of acid phosphatase was reduced in the macrophages in crush-syndrome. PMID- 18411725 TI - [Morphometric parameters of rat adrenals in the dynamics of general hypothermia]. AB - In this study, some morphometric parameters of adrenals in outbred albino male rats were compared in the dynamics of general hypothermia at the ambient temperature of -18 degrees C. It was shown that in the course of general hypothermia, the increase of blood vessel relative volume in zona reticularis and in adrenal medulla was accompanied by the augmentation of nucleus size of adrenocorticocytes (mainly in zona fasciculata). Zona fasciculata reacted with a significant increase of blood vessel relative volume during the 1st hour of cold exposure, henceforth the parameters remained unchanged. Blood vessel relative volume in the left adrenal was found to significantly exceed that in the right adrenal. In zona glomerulosa of the left adrenal, blood vessel relative volume was reduced, while that one in zona glomerulosa of the right adrenal remained unchanged during the whole experiment. Volume density of mitochondria in the endocrine cells of zona fasciculata was found to increase, this effect being more pronounced in the right adrenal as compared to the left one. In the cells of both glands, the volume density of lipid inclusions was gradually reduced, while the relative volume of nucleoli was variable and there were no statistically significant changes detected during the course of hypothermia. PMID- 18411726 TI - [Biogenic amine level in the normal placental structures and in antiphospholipid syndrome]. AB - Using the luminescent-histochemical methods, placenta was studied in 50 normal women with physiological pregnancy and in 35 patients in which the pregnancy progression was complicated by an antiphospholipid syndrome. The following bioamine-positive structures were identified in placenta: decidua, connective tissue and the blood vessels of chorionic plate, terminal villi containing capillaries, and syncytiotrophoblast. In antiphospholipid syndrome, the increase in histamine, catecholamine and serotonin luminescence intensity was detected in these structures. This increase was especially pronounced in histamine luminescence intensity. PMID- 18411727 TI - [Morphological changes in the fetal part of the allantoic placenta in rats exposed to acute hypoxia]. AB - The aim of this study was to analyze the response of cyto- and syncytiotrophoblast elements of fetal part of allantoic placenta to general acute hypoxia and to determine their morphologic characteristics, which provide the basis for structural changes in placenta and for the disturbances in placental blood flow. Female Wistar rats on day 16 of gestation were exposed to hypoxia (8% oxygen concentration in the gas mixture) for 1 hr. Placenta was studied 1 and 3 days later. Short-term acute hypoxia during pregnancy was shown to induce an impairment of formation and maturation of the fetal part of placenta. In the labyrinth zone, apoptosis was enhanced and a significant number of hyperchromatic nuclei appeared in cyto- and syncytiotrophoblast, which lead to the loss of some cells of cytotrophoblast and resulted in the retardation of its development and transformation into syncytiotrophoblast. One day following hypoxia, a stimulation of adaptive processes was evident as demonstrated by a dramatic rise of mitotic activity in the cytotrophoblast of the labyrinth zone of placental fetal part. However, a delay of the labyrinth zone development persisted till the end of pregnancy. Number and volume of fetal blood vessels were decreased as compared to those in control. A sharp decline of number and size of sinusoids containing maternal blood as well as a significant decrease of blood volume in the sinusoids, suggest a spasm of the uterine and placental vessels causing an impairment of utero-placental blood circulation and development of placental insufficiency. PMID- 18411728 TI - [Reparative regeneration of calvarial bones]. AB - The patterns of reparative osteohistogenesis were studied using the model of defects of rabbit parietal bones (n=18) with light and scanning electron microscopy and morphometry. Posttraumatic regeneration of cranial calvarial bones was shown to include main stages, which were also characteristic for long tubular bones, such as: primary response to trauma, reparative osteohistogenesis and adaptive remodeling. In every stage, temporary cellular associations are functioning within the regenerate. These are regeneratory histions which are cellular effectors of reparative process based on the regularities of physiologic osteogenesis including the phases of resorption, reversion, formation and rest. By day 120 the injured area was shown to be filled predominantly with dense connective tissue, which is associated with functional adaptation without the action of physical load. PMID- 18411729 TI - [Craniological basis of operative approaches to the structures of posterior cranial fossa using endovideo-monitoring]. AB - Cranioscopic and craniometric characteristics of posterior cranial fossa and correlations between them were studied using 127 skulls with different cranial shape (dolicho-, meso- and brachicraniums). It was found that most of the craniometric characteristics were independent on gender and shape of the skull, while each characteristic had some individual peculiarities. Endovideomonitoring was used to assess the optimality of suboccipital paramedial and retrosigmoid approaches to posterior cranial fossa using 20 heads of the corpses belonging to adult individuals. It was demonstrated that retrosigmoid approach was optimal for the accessibility of major anatomical structures of posterior cranial fossa. PMID- 18411730 TI - [Localization of bcl-2 antiapoptotic protein in the human placenta]. AB - The aim of this investigation was to study the distribution of bcl-2 antiapoptotic protein in various structures of human placenta using immunocytochemical methods. Syncytiotrophoblast was found to have the greatest bcl-2 immunoreactivity, while cytotrophoblastic elements, connective tissue cells of the villi and the blood vessel walls were bcl-2-negative. It is suggested that accumulation of bcl-2 in the syncytiotrophoblast may reflect its high resistance to detrimental factors. PMID- 18411731 TI - [Application of hematoxylin in histological technique]. AB - This review of the literature presents the data on the history of development of the methods of staining of histological sections using the natural dye, hematoxylin. The content of solutions of the most commonly used alum hematoxylins is described. The information is given on the modern staining methods, in which alum and iron hematoxylins are used, as well as on the hematoxylins, in which vanadium, lead, bismuth, uranium and other metals' salts are applied as mordants. PMID- 18411732 TI - [Konstantin Aleksandrovich Grigorovich (1905-1996)]. PMID- 18411733 TI - [Clinical significance of carriage of rare variants of connexin-26 genetic polymorphism in gastric cancer]. AB - The authors present first results of investigations of the connexin-26 gene in DNA obtained from peripheral blood of 55 patients operated on for gastric cancer. Gastric cancer patients were found to have carriage of the Cx 26 gene that was reliably associated with the invasive ability of the tumor. Change of the connexin-26 gene in gastric cancer is evidence of an important role of intercellular gap junctions in the arising and development of gastric cancer. PMID- 18411734 TI - [On the nature of secondary ulcers of the stomach]. AB - The authors have studied two main groups of causes of the arising of secondary ulcers of the stomach: ulcerous aetiology due to stenosing duodenal ulcers (238 patients--81.1%) and non-ulcerous aetiology (53 patients--18.2%). It was found that both variants of secondary ulcers were characterized by a considerable part of so called "difficult" ulcers of the stomach and high bias towards bleedings. Operation of choice for secondary ulcers of the stomach must be resection of the stomach preferably by the Billroth-I method or application of termino-lateral anastomosis. PMID- 18411735 TI - [Intraoperative mapping of lymph outflow tracts in colorectal carcinoma]. AB - Based on an analysis of 60 cases the authors have shown that identification and target investigation of signal lymph nodes using intraoperative staining with special dyes allows correct assessment of the stage of malignant disease. The method also allows treatment of the patient with an adequate, corresponding to up to-date standards scheme of combined and complex treatment, gives possibilities of exact intraoperative diagnosis of lymphogenic metastasis and, as a result, allows extended operative intervention up to radical volumes. PMID- 18411736 TI - [Surgical treatment of atherosclerotic lesions of the terminal part of the aorta and iliac arteries]. AB - The operations of semiclosed endarterecromy of the aortofemoral zone arteries were shown to have a number of advantages over shunting interventions with synthetic transplants. They are quick, not traumatic and have better results of long term patency. However, their application is limited by certain indications. Femoral amputations in patients with lesions of the aorta and iliac arteries independent of the age should not be recommended as a method of treatment of critical ischemia of lower extremities. Reconstructive operations on the vessels of this zone provide longer duration and better quality of life of the patients. PMID- 18411737 TI - [Specific complaints and pain syndrome in nonspecific osteomyelitis of the spine]. AB - The complaints were studied and pain syndrome estimated in 125 patients with nonspecific osteomyelitis of the spine (NOS). Complex conservative treatment was given to 74 of the patients, on 51 patients radical-restorative operations on the spine were fulfilled. The investigation has revealed specific features of pain syndrome in acute and chronic forms of NOS, in operative and conservative treatment. PMID- 18411738 TI - [Potentialities of videothoracoscopy in diagnostics and treatment of traumatic hemothorax]. AB - Therapeutic-diagnostic videothoracoscopy was used in complex treatment of hemothorax in 86 patients for isolated and combined stab-slash wounds and closed traumas of the chest. Thoracoscopy was not used in 321 patients with the same pathology taken as control. It was found that videothoracoscopy used for medium and big hemothorax always allowed the doctors to make the precise topical diagnosis of chest injury and due to medicinal endosurgical manipulations the thoracotomies were made 2.1 times more rarely, the duration of pleural drainage was 2.2 times shorter. Complications of the traumatic disease were 1.6 times less often, the time of in-patient treatment and lethality 1.3 times less. PMID- 18411739 TI - [Injuries of the duodenum]. AB - Based on an experience with treatment of 68 patients with injuries of the duodenum the authors have developed a diagnostic algorithm always permitting the diagnosis of injuries of the retroperitoneal part of the duodenum. A classification of severity of the duodenal injury was proposed determining the volume and type of surgery. Using the proposed therapeutic-diagnostic program decreased postoperative lethality from 52.4% to 11.8% in patients with wounds and from 60% to 13.4% in patients with closed injury of the duodenum. PMID- 18411740 TI - [The Troyanov-Trendelenburg operation in combination with sclerotherapy as an effective method of outpatient treatment of varicose disease]. AB - The authors have analyzed the strategy and results of treatment of varicose disease of lower extremities in 452 patients on an outpatient basis. It was shown that the Troyanov-Trendelenburg operation should be used as the first stage of treatment, and sclerotherapy should be carried on after the 4th day. PMID- 18411741 TI - [Outpatient treatment of patients with fractures of hand tubular bones]. AB - Results of treatment of 282 patients with 288 fractures of tubular bones of the hand were analyzed. In 190 (67.4%) cases conservative methods of treatment were used, in 92 (32.6%)--surgical methods. In cases of fractures without displacement plaster immobilization was used, in patients with stable fractures of finger phalanx and metacarpal bones with fragment displacement we made closed reposition and plaster immobilization. We applied intramedullary osteosynthesis with Kirschner's wire after closed reposition in unstable oblique and transverse fractures with fragment displacement. In open fractures of distal phalanx thin wires were used for intramedullary osteosynthesis. External fixation with Ilizarov's instrumentation was applied in patients with intraarticular fractures, with fractures healed in improper position and in patients with posttraumatic bone deformity. Using the surgical method in out-patient practice reduced the amount of hospitalized patients. Special methods of rehabilitation decreased the period of management and earlier returned the patients to active life. PMID- 18411742 TI - [Dysfunction of the endocrine system in patients with abdominal sepsis]. AB - The article presents results of a prospective investigation of 270 patients with abdominal sepsis. Three variants of endocrine impairments were established which correlated with the intensity of syndrome of inflammatory response and severity of the patients' state. In addition, possible transition of one variant of the endocrine dysfunction into another was proved in case of increased number of programmed sanitation relaparotomies. PMID- 18411743 TI - [Videothoracoscopic vagotomy in surgical treatment of patients with peptic ulcer of the gastroenteroanastomosis after resection of the stomach]. AB - Treatment of 130 patients with peptic ulcer of the gastroenteroanastomosis after resection of the stomach included operation by the method of videothoracoscopic truncal vagotomy (22 patients), left-side videothoracoscopic truncal vagotomy (19 patients), and operation from the right-side access (3 patients). Cicatrization of the ulcer in the postoperative period was obtained in 16 patients. Recurrent peptic ulcer was revealed in 2 patients. There were no lethal outcomes. PMID- 18411744 TI - [Experience with using of solutions of hydroxyethyl starch (refortan and voluven) in neuroanesthesiology]. AB - The investigation has shown that solutions of hydroxyethyl starch included in complex infusion therapy facilitate stabilization of hemodynamics in neurosurgical patients during surgery. Refortan exerts a more pronounced effect on the indices of hemodynamics as compared with voluven, but its modifying action on the hemostasis system is also more pronounced. These medicines used in doses 6 8 ml/kg fail to have substantial effects on efficiency of hemostasis in the brain wound and do not increase risk of postoperative hemorrhagic complications. PMID- 18411745 TI - [Effects of efferent methods of treatment on hemostasis in patients with postinfarction aneurysms of the left ventricle]. AB - Surgical operation of coronary shunting with resection of postinfarction aneurysm of the left ventricle (PALV) had to be done on 65 patients. Automatic plasmapheresis (AP) and plasmothrombocytopheresis (PTCA) were performed in the reoperation period for transfusion maintenance. The cellular composition of peripheral blood was not changed under the influence of cellular operations with the exception of thrombocytes: after PTCA their content was reliably decreased. AP and PTCA operations in patients with PTLV led to initially increased content of blood fibrinogen, whole protein in 24 hours, but by the time of operation the indices of protein metabolism were stabilized at the initial level. The operations of AP and PTCA were followed by a reliably decreased concentration of cholesterol, beta-lipoproteins and triglycerides that was associated with plasma exfusion. PA and PTCA fulfilled before operation in PALV patients allow both reservation of autologous plasma and thrombocytes and normalization of certain indices of hemostasis. PMID- 18411746 TI - [Correction of microcirculation in patients with atherosclerotic lesions of different vessels by efferent methods]. AB - The state of microcirculation was studied in 35 patients with obliterating atherosclerosis of the lower extremity vessels (1st group) and 36 patients with extensive atherosclerosis having a combined impairment of the peripheral basin: arteries of lower extremities and coronary arteries (2nd group) treated using the method of plasmapheresis (PPh) and PPh combined with intravascular laser irradiation of blood (IVLIB). The isolated use of PPh in patients of the 1st group was established to be effective, while combination of PPh and IVLIB was necessary for the patients of the 2nd group. PMID- 18411747 TI - [Sternolaparotomy as a variant of operative access in thoraco-abdominal wounds under conditions of a central regional hospital]. PMID- 18411748 TI - [Extraneural metastasizing glioblastoma]. PMID- 18411749 TI - [Panniculitis as a rare complication of panaritium]. PMID- 18411750 TI - [Strategy of the emergent diagnostics and therapy of hemolytic posttransfusion complications]. PMID- 18411751 TI - [Unsolved questions of prosthesis of the anterior abdominal wall in hernias (part 2)]. PMID- 18411752 TI - [Ultrasonic diagnostics of closed traumas of organs of the abdominal cavity and retroperitoneal space]. PMID- 18411753 TI - [The growth and differentiation of root cap columella cells and the proper root grown in the stationary conditions and under clinorotation]. AB - The results of light- and electron-microscopic investigations of root apices of Beta vulgaris 3-day-old seedlings grown in the stationary conditions and under clinorotation are presented. It was shown that ultrastructure and topography of organelles in root cap statocytes (graviperceptive cells) and in the cells of distal elongation zone clearly reflected the different direction in their growth and differentiation in space and time in dependence on specialization and functions. Cell growth and genetically determined differentiation occur similarly to control, although certain differences in ultrastructure are evident on metabolism changes. PMID- 18411754 TI - [The effects of alien chromosomes on common wheat resistance to biotrophic fungal pathogens]. AB - Effective, perhaps new genes of resistance to brown rust and powdery mildew expressed in common wheat genome in plant ontogenesis were revealed on additive chromosomes of Aegilops searsii, Aegilops longissima, Secale montanum and Elytrigia elongata. PMID- 18411755 TI - [Stable expression of promoterless bar gene in transgenic rape plants]. AB - Phosphinothricin resistant plants of two rapeseed (Brassica napus L. var. oleifera DC.) spring industrial cultivars were obtained by Agrobacterium tumefaciens leaf disk transformation. Vector constructions contained the promoterless coding sequence of phosphinothricin acetyltransferase (bar) gene located between two inverted lox-sites (elements of Cre/lox recombination system of P1 phage) and selective neomycinphosphotransferase II gene (nptII). Integration of the alien genes was confirmed by the PCR analyses. Stable and linked inheritance of foreign genes in T1 and T2 progeny was shown. PMID- 18411756 TI - [Cytogenetic variation of Ungernia victoris cell lines during cultivation on culture media of different composition]. AB - Chromosome numbers of U. victoris cell lines obtained from the same bulb and cultured for a long time on different agar-solidified and liquid nutrient media differed significantly. The components of the nutrient media including phytohormones did not influence the ratio of cells with different ploidy levels in various lines while transfer of the calluses to the liquid media resulted in the increase of diploid metaphase frequencies. PMID- 18411757 TI - [Mutations induced by X-rays and some chemical reagents changing Drosophila melanogaster life span]. AB - It has been shown that most of Drosophila melanogaster mutant lines obtained as a result of X-rays irradiation (XI) as well as of the combined action of XI and some chemical agents are characterized by decreased indexes of average (7-40 %) and maximal (1-35 %) life span. Insertion-excision processes at the instable genes white and cut are among the reasons of decreased vitality and shortened life span in induced mutants. Collection of neurodegenerative mutants has been obtained under the influence of ENU. Fast dying of flies and decreased vitality correlated with time point of neurodegenerations in brain structure. PMID- 18411758 TI - [Influence of genetic variation of oaks as forage substrate on the fitness components of green oak leaf roller]. AB - The article is focused on the study of influence of genetic variation in DNA (RAPD-PCR, primer OPA 14) of oaks (Quercus pubescens L. and Q. petraea L.) on the variation of some fitness-linked traits in different genotype classes of the Tortrix viridana adults. The results show that the leafroller individuals from the same genotype class may possess the maximum fitness (fecundity and body size components) on the trees of one genotype class and the minimum fitness on the trees of another genotype class. The interaction of the "oak genotype"- "leafroller genotype" factors results in rise of the ties which are expressed in statistically reliable associations between the leafroller and oak genotype classes. The results are discussed from the point of view of the newly developed field of community or ecosystem genetics. PMID- 18411759 TI - [The effects of actovegin on cell proliferation of permanent lines]. AB - The influence of Actovegin on proliferation activity and mitotic regimen of cells of permanent lines PK-15-IEKVM and BHK-21 clone 13/04 was investigated. Addition of Actovegin into growth media containing bovine serums of different components and concentrations stimulates cell proliferation. Conclusion has been made that Actovegin can be used in cell culture biotechnology. PMID- 18411760 TI - [Cytogenetic abnormalities in mammalian somatic cells]. AB - The different types of cytogenetic abnormalities are considered which are used in classic cytogenetics for the estimation of the levels of chromosome apparatus damages. The possible causes of cytogenetic anomalies and a number of methods of micronucleus tests are discussed. It was shown that the different levels of genetic material organization influence the realization of DNA defects into cytogenetic abnormality. PMID- 18411761 TI - [Interaction of RNA-containing bacteriophages with the host cells: MS2-induced E. coli mutants and formation of DNA-containing derivatives of MS2 bacteriophage]. AB - Sensitive cells of Escherichia coli AB 259 Hfr 3000 infected with RNA-containing phage MS2 produce phage particles and continue to divide showing segregation of sensitive cells maintaining new infection cycles. Phage multiplication in sensitive cells gives rise to phage resistant forms in their progeny. The described phenomenon has been shown to be due not to pre-existing phage-resistant cell selection but is a result of interaction of the phage and the cell. In contrast to the usual spontaneous or chemically induced Escherichia coli mutants MS2-induced phage-resistant cells are genetically unstable. During their reproduction they segregate new MS2-resistant types carrying more significant changes in the region coded by the sex factor. Cells belonging to two final MS2 induced mutants also produce a new type of phages; they are DNA-containing forms neutralized, however, by anti-MS2 serum. Production of such phage proves that genetic moiety of RNA-containing phage is able to be expressed as a part of the DNA structure. PMID- 18411762 TI - [Which infectious diseases should be prioritized in educating Japanese population?]. AB - OBJECTIVE: We studied which infections would be prioritized in educating the general population of Japan. MATERIALS AND METHODS: Subjects were 25 physicians and veterinarians in charge of infection control in infections control divisions of local and national governments. We conducted a questionnaire using the Delphi method. RESULTS: Based on (1) epidemiological and clinical characteristics, (2) knowledge level, awareness, and behavior of general population and healthcare professionals, and (3) social background and coping skills we selected 24 diseases for prioritization Tuberculosis was first, followed by influenza, HIV/AIDS, enterohemorrhagic Escherichia coli infection (O157), and genital chlamydial infection. Three animal-derived infections ranked in the top 10. DISCUSSION: We have not yet found which the priority of infections should be prioritized in educating the general population on infections. Although our findings are too few to make any generations about, several studies back the reasons why diseases were singled out in our study. Given the fact that most high ranked infections have ever been educated, education thus far appears to have been highly ineffective way, meaning that more effective ways of education on infection must be found in future. PMID- 18411763 TI - [Case of pneumothorax associated with pulmonary Mycobacterium fortuitum infection]. AB - A 39-year-old man with dyspnea was revealed to have severe pneumothorax and received partial resection of the left upper lobe after unsuccessful drainage. Necrotizing epitheloid granuloma was found in the resected lung and Mycobacterium fortuitum was detected from the lesion. Chemotherapy with levofloxacin and clarithromycin was started one year after surgery because of the newly found nodular shadow near the lesion. The case experienced pyothorax due to pulmonary tuberculosis three years before and Mycobacterium avium pleuritis one year before this episode. Three-time mycobacterial pleural infection in three years seems to be uncommon. Furthermore this is the first report of pneumothorax associated with pulmonary Mycobacterium fortuitum infection. PMID- 18411764 TI - [Fulminant Japanese spotted fever--the second fatal case in Japan]. AB - A 77-year-old woman who have no past history, was admitted in a local hospital in Muroto City, Kochi, Japan, after several days of fever and severe general fatigue and generalized skin erythema. She was suspected to have Japanease spotted fever, which was a local pandemic disease. She was treated with minocycline immediately. The next day, she had consciousness disturbance and low blood pressure. Laboratory findings indicated disseminated intravascular coagulation (DIC) and multiple organ failure. She was referred to our hospital. An eschar was identified in the back of It. femur. Treatment included minocycline, ciprofloxain, gabexate mesilate, methylprednisolone, hemodialysis and mechanical ventilation. In spite of the avobe treatment, she died 3 days after admission of the local hospital. Though the serological test showed no positive antibody titer against Rickettsia japonica, Rickettsia japonica was isoleted from blood culture, to confirm Japanese spotted fever, Japanese spotted fever is generaly a curative disease with early diagnosis and minocycline. In this case, the patient died 3 days after proper diagnosis and treatment was started. We reported the second fatal Japanese spotted fever case in Japan. PMID- 18411765 TI - [Case of Scedosporium apiospermum cutaneous soft tissue infection treated with voriconazole]. AB - A 69-year-old man treated with corticosteroids and immunosuppressive agents for acutely exacerbated interstitial pneumonia was found to have an ingrown nail in the left big toe and that suppurated despite treatment by dermatologists. Culture of the pus expressed from the toe yielded Scedosporium apiospermum. The patient suffered liver dysfunction a few days later when treated with intravenous voriconazole (VRCZ), which was discontinued due to the high plasma VRCZ concentration. Discrete erythema and subcutaneous nodules developed in left leg 2 or 3 weeks later. Ultrasonography showed tubular structures with substantial echoes that were not connected to veins in the subcutaneous tissue of the left leg. These findings suggested a nodular lymphangitic pattern of spreading of S. apiospermum soft tissue infection. Oral VRCZ at 100 mg/day was started, and increased to 200 mg/day after the plasma VRCZ concentration was measured. VRCZ was stopped after about 2 months, by which time the man had fully recovered. Because VRCZ-induced liver dysfunction was reported significantly associated with plasma level, we treated this case safely by administering VRCZ while measuring the plasma concentration. PMID- 18411766 TI - [Yersinia psuedotuberculosis septicemia in a healthy young woman]. AB - We report a case of Yersinia pseudotuberculosis (Y. pseudotuberculosis) septicemia in a healthy 22-year-old woman with clinical manifestations including high fever and general fatigue. The patient's ferer sudden elevation of her fever, and, liver damage and elevated inflammatory markers were indicated. General fatigue and appetite loss were noted on hospitalization. Exanthema was recognized, and all oral medications, including antibiotics, was stopped. The fever continued and high inflammatory parameters developed. After chemotherapy with imipenem, subjective symptoms ameliorated. Membranous desquamation without itching appeared between the fingers of both hands but improved naturally. Y. psuedotuberculosis septicemia was diagnosed as Y. psuedotuberculosis isolated from blood and elevated serum antibody titer against Y. psuedotuberculosis 4b detected at 1:160. PMID- 18411767 TI - [Case of multiple vertebral osteomyelitis due to Streptococcus pneumoniae]. AB - We report a rare case of multiple vertebral osteomyelitis due to Streptococcus pneumoniae. A 73-year-old man admitted for back pain and a low-grade fever was found in laboratory studies to have severe leukocytosis and increased C-reactive protein, but neither computed tomography (CT) nor vertebral magnetic resonance imaging (MRI) clarified the cause of infection in the painful hip lesion, and paralysis developed. in the left leg MRI eventually indicated a vertebral abscess involving multiple lesions at C4-7 and L4-5. We had started antibiotics before blood culture clarified Streptocccus pneumonaie, and antibiotics acted more effectively thereafter. The clinical course was good, little paralysis remained. PMID- 18411768 TI - [Use of the buccal fat pad for vocal fold injection]. AB - In recent years, autologous fat has been increasingly used for intracordal injection as a means of vocal rehabilitation in patients with deficient glottal closure, and the dipose tissue used has for the most part been harvested from the lower abdomen. However, patients differ considerably in the amount of fat available in the lower abdomen, and obtaining sufficient fat is often difficult in patents with a low body mass index (BMI). To overcome this problem we tried adopting a method that uses the buccal fat pad, as practiced in plastic surgery. The details of the method and the postoperative course are reported. The intracordal fat injection method for buccal fat pad tissue was performed in 10 cases during the 1-year from January 2005 to December 2005, and the patients' postoperative course was carefully monitored for more than 6 months after surgery. With the exception of one case, no signs of complications, such as swelling, infection, etc., were seen at the site of fat collection or in the vocal cord after the operation. Phonation was markedly improved after the injection, and the improvement was maintained throughout the observation period. Treatment of unilateral recurrent nerve paralysis by intracordal injection of autologous fat harvested from the buccal fat pad was successful. PMID- 18411769 TI - [Preoperative embolization and postoperative complications of carotid body tumors]. AB - Carotid body tumors are uncommon neoplasms that arise at the bifurcation of the common carotid artery. Surgical resection is generally recommended, but entails an inherent risk of cranial nerves injury and excessive blood loss. Preoperative embolization has been reported to decrease blood loss and shorten resection time. In this study, we analyzed the benefits of preoperative embolization and the postoperative complications when preoperative embolization was performed. Six patients with seven tumors were treated between 1990 and 2005. Each patient's preoperative evaluation included CT, MRI, US, and angiography, and preoperative embolization was performed in four patients with five tumors. Blood loss, in the patients who underwent preoperative embolization, ranged from 20 ml to 900 ml (mean: 291ml), and operation time ranged from 4 hours 34 minutes to 6 hours 40 minutes (mean: 4 hours 55 minutes). In the group that did not undergo preoperative embolization, blood loss ranged from 642 ml to 1390 ml (mean: 1016 ml), and operation time ranged from 9 hours 48 minutes to 10 hours 45 minutes (mean: 10 hours 17 minutes). Five patients had postoperative cranial nerve dysfunction, and it involved cranial nerve IX in one patient (14.3%), cranial nerve X in two patients (28.5%), and cranial nerve XII in five patients (71.4%). Resection of bilateral carotid body tumors in one patient resulted in baroreflex failure syndrome. In conclusion, preoperative embolization tends to decrease blood loss and shorten operation time, resulting in lower postoperative neurologic morbidity. PMID- 18411770 TI - [Efficacy and safety of propiverine on improvement of symptoms and QOL for female patients with wet overactive bladder]. AB - The present study was prospectively conducted for female patients with wet overactive bladder to assess the efficacy of propiverine hydrochloride in improving their symptoms and quality of life (QOL). Propiverine hydrochloride was administered orally to 58 patients for 8 weeks. Prior to administration and at 4 and 8 weeks after administration, symptoms and QOL were assessed by the micturition diary, the International Consultation on Incontinence-Short Form (ICIQ-SF), and the King's Health Questionnaire (KHQ). After administration, numbers of daily micturitions, incontinence episodes, urgency episodes and severity of urgency which were assessed based on the micturition diary and urgency scale questionnaire showed significant improvement when compared to baseline values. Furthermore, both ICIQ-SF and KHQ scores improved significantly after administration. The incidence of adverse events was 23.8% and none were serious. Propiverine hydrochloride was shown to contribute not only to the improvement of symptoms in female patients with wet overactive bladder but also to their QOL. PMID- 18411771 TI - [Clinical evaluation of supplemental administration of flavoxate hydrochloride in benign prostatic hyperplasia patients with nocturia resistant to an alpha1 adrenoceptor blocker]. AB - We examined the effectiveness of supplemental administration of flavoxate hydrochloride in patients with benign prostatic hyperplasia (BPH) whose nocturia was not adequately relieved by an alpha1-adrenoceptor blocker. Fifty-two patients who had two or more nocturnal micturition after administration of tamsulosin hydrochloride or naftopidil for 4 weeks or more received 400-600 mg of flavoxate hydrochloride in addition to an alpha1-adrenoceptor blocker for another 8-12 weeks. With supplemental administration of flavoxate hydrochloride, significant improvement was observed in the number of nocturnal micturition, total International Prostate Sympton Score, quality of life score and BPH impact index. No significant change was observed in the voided volume, Qmax, voiding time and residual urine volume. Supplemental administration of flavoxate hydrochloride is therefore effective for the improvement of nocturia and QOL in BPH patients resistant to an alpha1-adrenoceptor blocker. PMID- 18411772 TI - Changes in smooth muscle cell phenotype and contractile function following ischemia-reperfusion injury in the rat urinary bladder. AB - Twenty-eight adult male Sprague-Dawley rats were divided into four groups: Group 1 received 1 hour (h) of bilateral ischemia alone. Groups 2 and 3 received 1 h ischemia followed by 1 and 4 h of reperfusion (I-R), respectively. Group 4 consisted of age-matched control rats. Bladder strips were studied using electrical field stimulation (EFS) and KCl stimulation. Maximal contractile responses were recorded and analyzed. Temporal patterns of changes in phenotypic (non-contractile and contractile) expression of bladder smooth muscle cells were investigated using electron microscopy. The mean ratio of non-contractile to contractile phenotype (nc/c) of smooth muscle cells (SMCs) in the control group was 0.169. In the ischemia alone group, the ratio was 0.991. In the 1 h I-R group, the ratio 0.865 whereas in 4 h I-R group the ratio 1.601. The contractile responses to EFS and KCl showed decreased responses in all groups. These results clearly demonstrated that the ratio of nc/c increased significantly in the ischemia group and further increased significantly in both I-R groups. The contractile responses decreased in all ischemic groups although the magnitude of the contractile changes did not correspond in the change of phenotype ratio. PMID- 18411773 TI - [Clinical characteristics in patients with purple urine bag syndrome]. AB - Purple urine bag syndrome (PUBS) occurs predominantly in chronically catheterized and constipated patients. This syndrome is associated with bacterial urinary tract infections that produce sulfatase or phosphatase. Tryptophan is converted to indole and indigo-producing bacteria have indoxyl phosphatase or sulfatase that can produce indigo (blue) and/or indirubin (red) in patients with urinary tract infection. To further explore the metabolism of these amino acids, we evaluated the serum levels of amino acids in patients with PUBS. A total of 15 patients were enrolled in this case-control study (PUBS-positive case group: 5 patients, PUBS-negative control group: 10 patients). Data from urine tests (pH, sugar, protein, leukocyte counts), renal functions (BUN, creatinine), serum levels of amino acids, and performance status were compared between the two groups. No significant differences were seen between the two groups in urine sugar, protein, leukocyte counts, renal functions, and performance status. The mean urine pH was significantly higher in PUBS patients than in control patients (8.5+/-0.0 vs. 7.3+/-1.16, respectively, p=0.0321), and serum levels of alpha aminobutyric acid were significantly higher in PUBS patients than in control patients (16.2+/-3.08 vs. 12.4+/-3.20, respectively, p=0.0275). These data suggest that strong alkaline urine acts as an important factor in PUBS, in combination with other facilitating factors. PMID- 18411774 TI - [Transurethral resection of prostate just following high intensity focused ultrasound in localized prostate cancer--trial for early removal of the urethral catheter]. AB - We studied the impact of combined transurethral resection of the prostate (TURP) and high intensity focused ultrasound (HIFU) for localized prostate cancer (CaP) to decrease side effects such as prolonged urinary voiding disturbance observed after HIFU treatment. Included in this study were 18 patients with clinically localized CaP indicated for HIFU just followed by TURP (TUR combination group). Complete response was defined in accordance with ASTRO consensus statement and negative sample in biopsies performed 6 months after the HIFU treatment. Prostate specific antigen (PSA) nadir, International Prostate Symptom Score (IPSS) and morbidity during follow-up of TUR combination group were compared with those of a control of 18 patients who took HIFU treatment alone (HIFU monotherapy group). No statistical significances on the values of preoperative parameters (PSA, prostate volume, Gleason score, and IPSS) between these two groups. The median follow-up duration was 10 (5-15) months in both groups. A statistically significant impact was observed between TUR combination group and HIFU monotherapy group on median catheter time (5 versus 13 days, P<0.0001), PSA nadir (0.096 ng/ml versus 0.430 ng/ml in median, P<0.05) and the evolution of the post-treatment IPSS (8 versus 13.5 in median, P<0.0003) at 3 months after treatment. Urethral stricture necessary for urethral dilation was noted in 1 patient (5.6%) in the TUR combination group while in 2 (11.1%) in the HIFU monotherapy group. CR was obtained in 88.9% in the TUR combination group and 83.3% in the HIFU monotherapy group. Our study suggests that the combination of TURP with HIFU treatment improves posttreatment urinary status without additional morbidity. PMID- 18411775 TI - [Five-year results of prostate specific antigen (PSA) screening in Yokosuka City- comparative study between PSA-screened group and non-screened group]. AB - A mass screening of subjects for prostate specific antigen (PSA) was conducted to investigate whether this method is effective in detection of early stage prostate cancer. From 2001 to 2005, 1022 patients in Yokosuka City with pathologically diagnosed prostate cancer by using prostate needle biopsy were divided into screened (S: 276 patients) and non-screened (NS: 746 patients) groups. Clinical factors (mean age, PSA at diagnosis, clinical stage, Gleason score, WHO classification, cases of radical prostatectomy) were evaluated and analyzed. Statistical significance was analyzed by Mann-Whitney's U-test. The mean age was 70.7 and 72.7 (p<0.0001) in the S group and NS group, respectively; mean PSA at diagnosis was 10.5 and 18.6 ng/ml (P=0.0139); percentage of organ-confined disease (T2b or lower clinical stage), 75 and 55 (p<0.0001); percentage of non metastatic diseases was 92 and 77 (p<0.001) and percentage of pathologically poor risk cancer (8 or more Gleason score) was 22 and 38 (p=0.0004), respectively. In conclusion, PSA mass screening was found useful to detect early stage prostate cancer in Yokosuka City. Further studies should be conducted to determine whether PSA mass screening will be able to decrease mortality of prostate cancer. PMID- 18411776 TI - [The efficacy of portable 3-dimensional ultrasound scanning device (Bladder Scan BVI6100) for measurement of residual urine volume--comparison with transabdominal ultrasound estimation]. AB - In 177 patients, we evaluated the efficacy of a portable 3-dimensional ultrasound scanning device (Bladder Scan BVI6100) in measurement for residual urine volume, compared with transabdominal ultrasound estimation, using values measured by urethral catheterization as actual values. A high correlation was demonstrated between values measured by BVI6100 and actual values. Error rates of the values measured by BVI6100 male mode, female mode and transabdominal ultrasound estimation to actual values were 42.7 +/- 32.6, 50.6 +/- 63.3 and 65.8 +/- 43.7%, respectively. The error rate of the values measured by BVI6100 female mode was significantly higher than that by BVI6100 male mode or transabdominal ultrasound estimation. Irrespective of patient sex, the error rate of the values measured by BVI6100 in male mode was significantly lower than that in female mode. Therefore, BVI6100 male mode was considered to be a useful alternative for measurement of residual urine with accuracy similar to that of transabdominal ultrasound estimation. PMID- 18411777 TI - [Histopathological characteristics of radical prostatectomy specimen and long term PSA changes in men with PSA levels of 4.0 ng/ml or less]. AB - Out of 132 prostate cancer (Pca) patients who underwent radical prostatectomy 31 (mean age 65 +/- 5 years) had prostate specific antigen (PSA) levels of 4.0 ng/ml or less (low PSA group). The average PSA level was 3.3 +/- 0.5 ng/ml in the low PSA group and 8.5 +/- 5.5 ng/ml in patients with a higher PSA (high PSA group). The pT2 ratio of the radical prostatectomy specimens was 74% (23/31) in the low PSA group and 55% (55/101) in the high PSA group, pT3a was 16% (5/31) and 31% (31/101), pT3b was 10% (3/31) and 10% (10/101), pN1 was 0% and 5% (5/101), respectively. The digital rectal examination (DRE) gave a positive result significantly (p = 0.026) less frequently in the low PSA group (6/31 : 20%), than in the high PSA group (44/101 : 44%). However all three pT3b patients with a low PSA were positive in DRE. This suggests the importance of DRE to detect significant Pca with PSA < or = 4.0. PSA was measured at least three times for more than one year in 19 of the 31 patients with a low PSA level before diagnosis. In 14 of these 19 cases (74%), PSA velocity was more than 0.5 ng/ml/ year and PSA doubling time was less than 4 years. Some patients with significant Pca can not be detected with a PSA cutoff level at 4.0 ng/ml. We recommend that individuals have their own PSA levels, and that long-term changes of PSA are sometimes very important to detect cases of Pca with lower PSA. PMID- 18411778 TI - [Retroperitoneal Castleman's disease presenting characteristic image finding]. AB - A 30-year-old woman was hospitalized in our institute with fever up and abdominal pain. Computerized tomography showed a retroperitoneal mass (8 cm in diameter) with arborizing calcification, which was enhanced homogeneously in the arterial phase. The operation was performed and parthological diagnosis revealed hyaline vascular type Castleman's disease. This characteristic calcification pattern is considered unique to Castleman's disease, and could be useful for future diagnosis. PMID- 18411779 TI - [Renal infarction with renal abscess onset due to infective endocarditis: a case report]. AB - A 68- year-old male was referred with fever and urinary tract infection. Abdominal computed tomography showed a right renal infarction with abscess and abscesses in the liver and spleen. Cardiac ultrasonic examination showed a vegitation in the aortic valve. Based on these findings, the patient was diagnosed with infective endocarditis and multiple abdominal organ abscesses. The renal infarctions and multiple abscesses probably developed due to dissemination of warts associated with infective endocarditis. We performed emergency valve replacement at the cardiovascular surgery unit at our hospital. Improvements were made in the multiple abscesses after subsequent treatment with an antibiotic agent. The renal diseases associated with infective endocarditis were found to be diverse and we concluded that it is important to understand the conditions of the disease correctly for appropriate treatment. PMID- 18411780 TI - [Bilateral adrenal metastases from renal cell carcinoma: a case report]. AB - We report a case of renal cell carcinoma with bilateral adrenal metastases. A 57 year-old man was admitted to our hospital for a left renal mass. Computerized tomography and magnetic resonance imaging revealed a 4.5 cm left renal tumor and bilateral adrenal masses (3.0 cm on the right side and 2.0 cm on the left). A left nephrectomy and bilateral adrenalectomies were performed. The pathological findings showed clear cell carcinoma, G2 of left kidney metastasizing to both adrenal glands. The patient was administered supplementary hydrocortisone and was well 8 months after the nephrectomy and adrenalectomies without evidence of recurrence. Bilateral adrenal metastases from renal cell carcinoma are relatively rare. Our case seems to be the thirteenth case in the Japanese literature. PMID- 18411781 TI - A case of renal cell carcinoma with intrahepatic vena caval tumor thrombus successfully managed by surgery in a long-term chronic hemodialysis patient with liver cirrhosis. AB - A 54-year-old man who had been under hemodialysis therapy for 16 years presented with gross hematuria at our department in February 2005. Imaging findings revealed right renal tumor of8.2 cm in diameter. In addition, the tumor extended into inferior vena cava at the level of the hepatic vein. There were no findings of distant metastasis. Right radical nephrectomy and thrombectomy were performed on April 2006. Histopathological analysis showed that the tumor was renal cell carcinoma of clear cell type, grade 2. Postoperative course was uneventful, and the adjuvant therapy with interferon alpha was initiated. He has been free from recurrence for 22 months after surgery. PMID- 18411782 TI - [A case of adenocarcinoma of the urinary bladder arising 45 years after ileal replacement of ureter for tuberculous ureteral stricture]. AB - We report a case of adenocarcinoma arising in the urinary bladder 45 years after ileal replacement of ureter for tuberculous ureteral stricture. A 65-year-old man was admitted with gross hematuria. Cystoscopic examination demonstrated a broadbased non-papillary tumor on the left posterior wall of the bladder and a papillary tumor at the anastomotic site between the bladder and ileal segment. Total cystectomy and construction of ileal conduit were performed. The histopathological examination demonstrated well differentiated adenocarcinoma of the bladder and ileal ureter. Intestinal metaplasia widely infiltrated into the bladder epithelium around the anastomotic site. PMID- 18411783 TI - [Successful treatment of hemorrhagic cystitis after radiation therapy with intravesical instillation of aluminium hydroxide gel and magnesium hydroxide: report of a case]. AB - A 77-year-old man was admitted to our hospital due to macrohematuria and bladder tamponade. Bladder hemorrhage caused by radiation therapy had not improved after bladder lavage, intravesical drip infusion, medication of hemostatics, and transurethral coagulation. Bladder hemorrhage had completely disappeared 7 days after the intravesical instillation of 50-100 ml aluminium hydroxide gel and magnesium hydroxide for an hour per day. This method is easy and can be performed without complications. This method might be useful as first-line therapy in the case of severe bladder hemorrhage. PMID- 18411784 TI - A case of prostatic duct adenocarcinoma: its clinical significance in comparison with typical acinar adenocarcinoma. AB - We report a case of prostatic duct adenocarcinoma treated with radical prostatectomy. Advanced pathological stage (pT3bpN1) was beyond the prediction of the favorable preoperative parameters (cT1cN0, PSA 7.64 ng/ml). The main tumor of ductal adenocarcinoma was occupying the transitional zone and surrounded by scattered micro-foci of acinar adenocarcinoma. We identified coexistence of ductal and acinar adenocarcinoma cells side by side in the same gland. Pure ductal cancer cells were detected in the metastasized lymph node without acinar cancer cells. Strong staining of PSA and loss of p63 expression by both types of adenocarcinoma cells were confirmed immunohistochemically. We discuss the clinical significance of prostatic duct adenocarcinoma in comparision with typical acinar adenocarcinoma. PMID- 18411785 TI - [Disagreement between cytology and biochemistry/molecular biology in studies of organelle inheritance--observation]. PMID- 18411786 TI - [Molecular organization of the membrane cytoskeleton revealed by electron microscopy]. PMID- 18411787 TI - [Cell adhesion and the cytoskeleton]. PMID- 18411788 TI - [Nuclear envelope assembly by endoplasmic reticulum during cell cycle]. PMID- 18411789 TI - [Chromosome assembly controls: perspective from the observation of human artificial chromosomes]. PMID- 18411790 TI - [Mechanisms of spindle pole formation for the proper chromosome segregation]. PMID- 18411792 TI - [Regulation of cell polarity and intracellular organelles in retinal photoreceptor cells]. PMID- 18411791 TI - [Key questions about membrane dynamics during autophagy]. PMID- 18411793 TI - [Coordination of actin cytoskeleton and membrane transport in cell migration]. PMID- 18411794 TI - [Neutron capture therapy for glioblastoma]. PMID- 18411795 TI - [Microvascular compression syndrome: review and update of microvascular decompression surgery]. PMID- 18411796 TI - [Facial nerve motor evoked potentials elicited by transcranial electrical stimulation for intraoperative monitoring]. AB - OBJECTIVE: To determine whether monitoring facial nerve motor evoked potentials (FNMEPs) elicited by transcranial electrical stimulation during skull base tumor surgery is useful for predicting facial nerve outcome. METHODS: We analyzed FNMEP findings in 31 patients with skull base tumor. Surgery was performed twice in 2 of the 31 patients because of tumor regrowth. FNMEP monitoring was conducted 33 times in the present study. Corkscrew electrodes positioned at C3 or C4 and Cz were used to deliver supramaximal stimuli (140-550V). FNMEPs were recorded from the orbicularis oculi and oris muscles. Correlation between the final-to-baseline FNMEP ratio and postoperative facial nerve function (House & Brackmann grade) was examined. RESULTS: Valid FNMEPs were obtained in 26 of the 33 (78.8%) recordings from the orbicularis oculi muscle and in 31 of the 33 (93.9%) recordings from the orbicularis oris muscle. Facial nerve function correlated significantly with the FNMEP ratios in the orbicularis oculi (r = -0.52 N=26, p < 0.01) and orbicularis oris (r = -0.60, N=31, p < 0.001) muscles. An FNMEP ratio of -50% consistently predicted immediate postoperative facial palsy, although the degree of palsy differed among the patients. CONCLUSIONS: Intraoperative FNMEP monitoring is useful for predicting facial nerve function after skull base surgery. PMID- 18411797 TI - [Acute brain swelling during operations for three patients with moyamoya disease resulting in performance of indirect anastomosis as a substitute for direct anastomosis]. AB - Surgery for the treatment of moyamoya disease includes direct and indirect anastomosis. In the present study, three cases with moyamoya disease, in which acute brain swelling in the operative field occurred just after opening of the dura mater and indirect anastomosis was performed as a substitute for direct anastomosis to terminate the operation earlier than originally planned. They all waked immediately from general anesthesia and showed no neurological deterioration. Computed tomographay of the brain performed just after the operation showed no abnormalities such as acute infarction, hemorrhage, global brain swelling, and local brain swelling. One of the three cases showed signs of having caught a cold before the operation, another manifested allergic rhinitis just before the operation, and the other had taken a long walk and experienced hyperventilation resulting in TIA associated with quadriparesis a day before the operation. Abrupt return from hypocapnea to normocapnea during the induction of general anesthesia was thought to be the cause of the acute brain swelling. It was thought desirable that correction of hypocapnea needed to be performed gradually during the operation in those cases. PMID- 18411798 TI - [Endosaccular embolization of an intrasellar aneurysm with hypopituitarism: case report]. AB - An 81-year-old male presented with weight loss and hyponatremia. A baseline endocrinological examination revealed anterior pituitary dysfunction. A computed tomography and MRI revealed a large, unruptured intrasellar aneurysm protruding from the right internal carotid artery and pushing the pituitary stalk leftward. The patient developed polyuria and polydipsia from 10 days after commencing corticosteroid supplementation and was diagnosed with partial central diabetes insipidus (DI). Balloon assisted endosaccular embolization was performed about one month after the posterior pituitary dysfunction was identified. The unruptured aneurysm was successfully embolized with Guglielmi detachable coils (GDCs) without narrowing of the parent vessel. DI resolved completely and the posterior pituitary dysfunction improved soon after endosaccular embolization. The remission of DI after coil embolization suggested that the DI might have been induced by the progressive mass effect of the aneurysm rather than by the steroid. An endocrinological evaluation and cerebral angiography confirmed partial recovery of anterior pituitary dysfunction and almost complete obliteration of the aneurysm, respectively at 1 year after the operation. We report a case of hypopituitarism secondary to the large intrasellar aneurysm. This aneurysm was embolized with GDCs, resulting in partial recovery of anterior pituitary dysfunction and complete recovery of posterior pituitary dysfunction. PMID- 18411799 TI - [Massive epistaxis due to traumatic internal carotid artery pseudoaneurysm: a case report]. AB - We present a case of traumatic pseudoaneurysm of the internal carotid artery (ICA) accompanied by skull base fracture, which was treated by endovascular trapping of the internal carotid artery. A 70-year-old woman met with a traffic accident and was admitted to our institution with epistaxis and accomapanied with shock. Angiography on day 2 showed carotidcavernous fistula. The patient had no eye symptom and was treated conservatively. A second angiography, two weeks aftrer the trauma, revealed development of a pseudoaneurysm on the C3-4 portion. We attempted balloon test occlusion (BTO) of the right internal carotid artery, and if torelated, the ICA may have been occluded. The day before BTO, she rebleed massively. Endovascular trapping of the ICA was performed. Although epistaxis was controlled completely, she suffered left hemiparesis due to an embolism during the procedure. Epistaxis from a traumatic aneurysm of the internal carotid artery may be fatal and emergency occlusion is proposed. PMID- 18411800 TI - [Intramedullary spinal cord metastasis treated with radiation therapy: report of 3 cases]. AB - OBJECTIVES: Intramedurally spinal cord metastasis (ISCM) tends to be seen in the end period of the malignant tumor, so it is important to choose a therapeutic method regarding the QOL (quality of life) of the patient. In this study, we reported three cases of ISCM treated by radiation therapy alone, and demonstrated the utility of this therapeutic procedure. PATIENTS AND METHODS: From 2005 to 2006, 3 cases of ISCM underwent radiotherapy in our institution. The mean age of the patients was 64.0 years old, and all patients were men, having lung carcinoma as their primary lesions. The lesions were located at the cervical cord in one case and the conus medullaris in the other two cases. They were treated by radiotherapy at the department of radiology in our institute. RESULTS: The tumor size and intramedullary high signal area on MR images after the radiotherapy were reduced in all patients. During the clinical course, neurological symptoms were improved in two patients, and progression of the symptoms was stopped in one patient. All of the patients obtained a better ADL due to the radiotherapy. CONCLUSIONS: Radiotherapy has an advantage over surgical treatment in that it involves less invasion. The treatment brought improvements of clinical symptoms and radiological findings in the patients with ISCM. So, radiotherapy could be used as one of the major treatments for patients with ISCM who desire a better QOL after treatment. PMID- 18411801 TI - [Genetic diagnoses and individualized treatment of gliomas]. PMID- 18411802 TI - [Characterization of a large deletion that leads to congenital factor XIII deficiency]. AB - Congenital factor XIII deficiency is a rare bleeding disorder that is inherited in an autosomal recessive manner with a frequency of 1 per 2 million individuals in the human population. In Japan, 53 cases of factor XIII deficiency were registered in the national survey for blood coagulation disorders in 2006. One hundred twenty-three cases were listed in the international Factor XIII Registry (http://www.f13-database.de/) by October 2007. The most frequent genomic abnormalities among the registered cases are point mutations; nucleotide deletions have been identified in only 16 cases. Most deletions are less than 20 bp; only 2 large deletions have been reported. However, detailed studies in either of these 2 cases have not been performed. We analyzed a case of congenital factor XIII deficiency. The patient is Japanese born to consanguineous parents, and his factor XIII A antigen and activity levels are both less than 10% of normal. The LA-PCR product for exons 4-6 of the factor XIII gene was 5 kb smaller than expected. The deletion is exactly 5984 bp long, including the entire exon 5. This finding suggests that the deletion caused a frameshift that produced a premature termination codon in exon 6. Deletions usually occur in repetitive sequences, but repetitive sequences were not found around this deletion. The semiquantified F13A1 mRNA level in the patient sample was only 1% of normal, and suggests that the mRNA surveillance system (nonsense-mediated mRNA decay) may be involved. PMID- 18411803 TI - [Study on usefulness of different APTT test kit in variable coagulopathy]. AB - A number of test kits are available for measuring activated partial thromboplastin time (APTT) and are used to screen for intrinsic coagulation reactions. However, results obtained with the same sample by different test kits often vary, causing confusion regarding potential hemostatic activity in the specimen. We investigated the usefulness of 6 different APPT kits, which utilize various phospholipids and activators, to detect prolonged clotting time in plasma from subjects with abnormal coagulopathy, including lupus anticoagulant(LA). In samples from subjects with intrinsic coagulation factor deficiencies and subjects taken heparin, the abnormal APTT detection ratio was high regardless of the kit used, thus any would be acceptable for measuring APTT in such patients. In contrast, that ratio in patients with von Willebrand disease was relatively low regardless of the kit, probably because factor VIII activities in those patients were slightly decreased. The ratio of detected subjects with LA and subjects taking warfarin varied among the APTT kits, however, those that utilized synthetic phospholipids were useful for the detection of LA. Our results suggest that an APTT kit should be selected according to the kind of disorder in the patient. Further, kits that employ synthetic phospholipids are useful for detecting abnormal coagulopathy in patients with intrinsic coagulation factor deficiencies and patients taken heparin, as well as for detection of LA. PMID- 18411804 TI - [Laboratory medicine for blood transfusion and transplantation medicine]. AB - We discussed the usefulness of routine technologies of laboratory medicine in blood transfusion and transplantation medicine. New parameters that can be measured by automated hematology analyzers have been clinically evaluated and proven to be useful so far. Based on our experience, detection systems for fragmented red cells (FRC), immature platelets (immature platelet function, IPF), and hematopoietic progenitor cells (HPC) are useful for the diagnosis of thrombotic microangiopathy, differential diagnosis of thrombocytopenia, and decision regarding the optimal timing to collect peripheral stem cells, respectively. Moreover, IPF were suggested to be an indicator of the platelet transfusion requirement. The establishment of non invasive assaying technology has been eagerly anticipated. We evaluated a hemoglobin measurement tool, and revealed that it might be applicable in predeposited, autologous blood donation. Some adverse transfusion reactions are related to neutrophil activation. Thus, we investigated the effects of serum from patients and blood donors, in the context of adverse reactions, on adhesion molecule expressions of neutrophils from volunteers using flow-cytometry. This kind of simple technology is expected to be useful in future studies to clarify the mechanisms and prevent adverse reactions. PMID- 18411805 TI - [Problems of pre-analysis in genetic testing]. AB - Genetic testing has been developed to confirm various disorders and is applied widely as a fast-growing diagnostic tool. Laboratories performing genetic testing are needed to ensure quality assurance. Accurate and precise testing using nucleic acid extracted from various samples is important as pre-analysis as the results can be affected by bias introduced by sample preparations. The assays estimate the purification and isolation of nucleic acid from samples. Pre analytic processes such as clinical sampling affected the outcome of genetic testing. Analysis of variance of gene expression revealed small but significant differences between handling methods. Great care has to be taken to measure pre analytic changes in gene expression. Internal quality control programs for genetic testing are also needed. Thus, well-controlled sample processing and storage conditions are critical for sensitive and potentially quantitative analysis of genetic testing. PMID- 18411806 TI - [Pre-analytical problems on assay conditions for blood midkine]. AB - We have established the assay conditions of midkine (MK) measurement, the reference intervals and evaluation for clinical significance of blood MK measurement. MK is a kind of cytokines and basic protein which is a heparin binding growth factor of various cells. The increase of MK expression suggests a prognostic value in early stage on cancers or inflammation. But significant problems in the MK measurement are alterations resulting from standing time and temperature instability after blood collection. Assay of MK was performed with solid phase human MK immunoassay recently developed sensitive enzyme linked immunosorbent method. The assay condition of MK was required to be separated immediately after blood sampling within 24 hrs at 4 degrees C or within 2 hrs at room temperature-standing. Plasma sample obtained with EDTA-2Na or citric acid Na, and serum obtained from plain tube container showed good results. Linearity was obtained up to 1500 pg/ml and repeatability and reproducibility were within 10% as CV%. The recovery of MK was 101.1+/-3.8% with 10 specimens ranged 97-105%. Addition of interfering substances showed no effect on assay results when hemoglobin, EDTA-Na, citrate and turbidity check, but conjugated bilirubin (over 0.68 mmol/l) and gave negative errors within 10% in the assay results and heparin gave negative errors. The reference interval was 550 +/- 160 pg/ml in healthy individuals serum. PMID- 18411807 TI - [The significance of preventing pre-analytical errors in the pathology laboratory to obtain accurate pathological and cytological reports]. AB - Pathological and cytological reports are unique, because the results are not numerical, and they may become the final diagnosis, having a direct bearing on further therapeutic decisions. Recently, the numbers of medical incidents/accidents, in which pathology technologists and/or pathologists have been involved, have been increasing. It is of concern that a lack of proper procedures in pathology practice may give rise to serious accidents affecting patients' lives. For accurate pathology diagnoses, highly skilled pathological technicians and well-knowledgeable pathologists are necessary but, in addition, precise quality assurance is essential. Efforts to prevent pre analytical errors are presented and discussed. PMID- 18411808 TI - [Pre-test blood samples at clinical laboratories--effects of temperature and duration of blood sample storage on measurements]. AB - Increases in lactate dehydrogenase (LD) were observed from 24 hours for samples stored at normal temperature, and were noted from 12 hours for those stored at room temperature. Potassium (K) increased from 3 hours for samples stored under refrigeration, and increased from 6 hours for those stored at normal temperature. For those stored at room temperature, this parameter decreased from 6 through 12 hours after blood sampling and increased from 24 hours. Inorganic phosphorus (P) increased from 36 hours for samples stored at normal temperature. For those stored at room temperature, this parameter decreased from 3 through 6 hours after blood sampling and increased from 24 hours. Increases in magnesium (Mg) were noted from 24 hours for samples stored at normal temperature, and were observed from 12 hours for those stored at room temperature. Increases in iron (Fe) were noted from 36 hours for samples stored at normal temperature, and were observed from 24 hours for those stored at room temperature. Blood urea nitrogen(BUN) increased from 24 hours for samples stored at room temperature. Creatinine (CRE) increased from 24 hours for samples stored at room temperature. Uric acid (UA) decreased from 36 hours for samples stored at room temperature. Serum glucose decreased from 6 hours for samples stored under refrigeration, decreased from 6 hours for those stored at normal temperature, and decreased from 3 hours for those stored at room temperature. Although it has generally been accepted that it may be good practice to store blood samples under refrigeration, there have been no established theories to prove this. The authors investigated the effects of the temperature and duration of blood sample storage on measurements and found that for some parameters, the measurements varied depending on the temperature and, duration of storage. No specific temperature for the storage of blood samples could be identified in all cases and accordingly, the personnel concerned should refer to the relevant materials and documents so as to identify an appropriate storage temperature. PMID- 18411809 TI - The curious lives of surrogates. PMID- 18411810 TI - NMC under scrutiny. PMID- 18411811 TI - 'Mental well-being is a health promotion priority'. PMID- 18411812 TI - 'It's not ageist to say older people's needs may differ from those of younger people'. PMID- 18411813 TI - Can you eat well at work? PMID- 18411814 TI - Report finds inequity in access to breast surgery services. PMID- 18411815 TI - Patient assessment part 5--measuring pulse. PMID- 18411816 TI - Communication in end-of-life cardiac care 2: skills. AB - This is a two-part unit on communicating with patients with end-stage heart failure. Part 1 identified the potential issues patients may face. This second part aims to enable nurses to develop an awareness of the skills needed to help patients identify and express their fears and anxieties. PMID- 18411817 TI - Island life sees Ailsa hit the heights. Interview by Jonathan Baillie. AB - Brought up in a small Shropshire village, Ailsa says she enjoyed most subjects at school, but particularly English, Geography and Chemistry. However, in a possible early pointer to her subsequent career, she says she had "always been interested in food"; her mother was trained as a home economics teacher and, from an early age, Ailsa herself wanted to be a chef. PMID- 18411818 TI - Deep cleaning--valid or publicity stunt? AB - While deep cleaning is not new--hospitals have undertaken deep cleans ward-by ward for some time, often following a "superbug" outbreak, Health Secretary Alan Johnson and Prime Minister Gordon Brown both emphasised on announcing the mandate that this was the first time all English NHS hospitals had been asked to deep clean their entire site, "restoring surfaces and fabrics to as close as possible as their original condition". PMID- 18411820 TI - Sensitive side shown in pounds 3.7m new build. AB - Heatherwood and Wexham Park Hospitals (HWPH) NHS Foundation Trust provides hospital services and over 550 beds to a population of 400,000 across East Berkshire and South Buckinghamshire. The nine-month refurbishment and fit out programme at Wexham Park to provide the new sterile services unit got under way in January; it is scheduled to begin operating this October, employing 30 staff. PMID- 18411819 TI - Facility lifts cardiac care to new level. AB - Notably favourable patient outcomes are being recorded at the new pounds 60 million Essex Cardiothoracic Centre, reflecting not only how the facility is being run but also the care, precision and openness with which it was designed, procured and constructed. PMID- 18411821 TI - Fire safety--the law and one's obligations. AB - Early January's dramatic fire at London's Royal Marsden hospital, during which 79 patients and 200 staff were safely evacuated, highlighted the vital importance to healthcare facilities, large and small, of having in place proper, well co ordinated fire evacuation procedures. PMID- 18411822 TI - Fire safety in the firing line. AB - Broadcast on 29 January, the File on 4 programme, titled: How safe are hospitals from fires?, was aired in the wake of the fire at London's Royal Marsden Hospital on 2 January. While 79 patients and 200 staff were safely evacuated, the program me's presenter Allan Urry said that, over a month later, all five of the hospital's operating theatres and its critical care unit remained out of action; the fire's cause is still being investigated. PMID- 18411823 TI - Hospital sanitation examined. AB - Important medical and technical advances have been made in recent years. Hospital planning has become more and more specialised and, therefore, more and more complicated. PMID- 18411824 TI - Medical supply units--what are you given? AB - With recent changes in legislation throughout the European Union (EU) and the publication of revised and up-to-date Department of Health, NHS and professional body guidance documents, the ever-increasing burden of compliance now falls firmly upon manufacturers and installers. PMID- 18411825 TI - Youngsters' views should inform design. AB - Children and adolescents' differing views on what constitutes a comfortable hospital environment are examined in a study by Sanaz Litkouhi, Dr. Farhang Mozaffar and Dr. S. Bagher Hoseini of the Iran University of Science and Technology, Tehran. PMID- 18411826 TI - Breathing life into family processes: introduction to the special issue on families and asthma. AB - This introduction to the special issue dedicated to families and asthma proposes that the study of asthma highlights general systems topics such as integration of individual needs into the group, developmental trajectories of risk and resilience, supportive and destructive patterns of interaction, and the cultural adaptation of family therapy. It briefly introduces papers in the special issue and concludes that asthma can serve as an exemplar in the study of family health due to its comorbidity with mental health problems, potential to affect multiple members of the family, disproportionate influence on low-income and minority families, and multiple avenues for intervention. PMID- 18411827 TI - Caregiver depressive symptoms and observed family interaction in low-income children with persistent asthma. AB - This study examined the relationship between caregiver depressive symptoms and observed parenting behaviors and family processes during interactions among 101 urban, low-income Africtan American families with children with persistent asthma. Caregivers (primarily female) were assessed on four dimensions (i.e., warmth/ involvement, hostility, consistent discipline, relationship quality) in three videotaped interaction tasks (loss, conflict, cohesion). The results indicated that increased depressive symptoms were significantly associated with lower warmth/involvement and synchrony scores and greater hostility scores during the loss and conflict tasks. In the total sample, the highest levels of hostility and the lowest levels of warmth/involvement were found for the conflict task; nevertheless, caregivers with moderate/severe depressive symptoms showed a significantly greater increase in hostility from the loss to the conflict task than caregivers with minimal/mild depressive symptoms. The findings highlight the salience of considering task content in family observational process research to expand our understanding of depressed and nondepressed caregivers abilities to modulate appropriately their behaviors and affect across various family interactions. Implications for improving asthma management for low-income children with persistent asthma are discussed, including the utility of multidisciplinary interventions that combine asthma education with family therapy. PMID- 18411828 TI - Testing the Biobehavioral Family Model in pediatric asthma: pathways of effect. AB - This study uses a laboratory-based multiinformant, multimethod approach to test the hypothesis that a negative family emotional climate (NFEC) contributes to asthma disease severity by way of child depressive symptoms, and that parent child relational insecurity mediates the effect. Children with asthma (n = 199; aged 7-17; 55% male) reported parental conflict, parent-child relational security, and depressive symptoms. Parent(s) reported demographics, asthma history, and symptoms. Asthma diagnosis was confirmed by clinical evaluation and pulmonary function tests, with disease severity rated by an asthma clinician according to NHLBI guidelines. Family interactions were evoked using the Family Process Assessment Protocol, and rated using the Iowa Family Interaction Rating Scales. Path analysis indicated a good fit of data to the hypothesized model (chi2[1] = .11, p =.74, NFI = .99, RMSEA = .00). Observed NFEC predicted child depression (beta = .19, p < .01), which predicted asthma disease severity beta = .23, p < .01). Relational security inversely predicted depressive symptoms (p = .40, p < .001), and was not a mediator as predicted, but rather an independent contributor. The findings are consistent with the Biobehavioral Family Model, which suggests a psychobiologic influence of specific family relational processes on asthma disease severity by way of child depressive symptoms. PMID- 18411829 TI - Family psychological factors in relation to children's asthma status and behavioral adjustment at age 4. AB - The objectives of this study were to determine whether family psychosocial factors influenced asthma development by age 4, and whether family factors and early wheezing illness were associated with behavioral adjustment at age 4. Participants were 98 children enrolled in an intervention study at 9-24 months and followed to age 4. Baseline evaluations assessed infants' respiratory illness severity, family psychosocial characteristics, and parental risk factors for asthma development. Active asthma categorization at age 4 utilized both parent report and objective data. Parents completed the Child Behavior Checklist (CBCL). Caregiver single-parent status, a composite of baseline family stresses, and early wheezing illness severity were associated with active asthma at age 4. The contribution of prenatal smoke exposure and early hospitalization to active asthma varied with racial/ethnic group membership. Maternal mental health and family stresses predicted CBCL scores at age 4, whereas early illness severity and hospitalization were unrelated to CBCL scores. CBCL scores were not elevated for children with active asthma at age 4. Family factors consistent with a negative emotional environment were associated with both active asthma and adjustment problems at age 4, suggesting that both outcomes may be influenced by a common factor. PMID- 18411830 TI - Family climate of routine asthma care: associating perceived burden and mother child interaction patterns to child well-being. AB - This preliminary report links the literatures on family asthma management practices and on the characteristics of family interaction patterns thought to influence children's adjustment to a chronic physical illness. Specifically, this study of 60 families with a child with asthma examined the extent to which perceived burden of routine asthma care affected child mental health via its influence on parent-child interaction patterns. Mothers completed a measure of asthma management routine burden, mother and child were observed in a 15-minute interaction task, and children completed measures of child anxiety and asthma quality of life (QOL). Perceived routine burden significantly predicted child anxiety and QOL through its effect on mother-child rejection/criticism. The same pattern did not hold for mother intrusiveness/control. The results are discussed in terms of how overall family climate and regulation of routines affects child well-being. Implications for clinical practice and limitations of the study are provided. PMID- 18411831 TI - Correlates of household smoking bans in low-income families of children with and without asthma. AB - Exposure to secondhand smoke (SHS) harms all children's health, especially children with asthma. Yet, children with asthma are as likely to live with smokers as healthy children. Household smoking bans are being advocated to reduce children's harm from SHS. To measure the effect of household smoking bans on child SHS exposure and to examine correlates of strict smoking bans in a low income, diverse sample, 91 children with asthma were matched to 91 healthy children. All had at least one smoker living in their homes. Nicotine dosimeters, child cotinine assays, and maternal reports quantified child SHS exposures. Maternal reports of household smoking rules, behaviors, and beliefs, and other family characteristics were also gathered. The presence of a strict household smoking ban vastly reduced children's SHS exposures and was associated with fewer cigarettes smoked by the mother and by other family members, the belief that SHS was a personal health risk, having children with asthma, and living in a single family home. Many children are exposed to high levels of SHS at home. Strict household smoking bans greatly decrease, but do not eliminate children's SHS exposure. Even in disadvantaged families, mutable factors were associated with strict smoking bans. Increased dissemination and use of established public health strategies are needed to reduce children's SHS exposures. PMID- 18411832 TI - Feasibility and impact of a school-based intervention for families of urban adolescents with asthma: results from a randomized pilot trial. AB - The purpose of this study was to test the feasibility and short-term outcomes of Asthma: It's a Family Affair!, a school-based intervention for adolescents with asthma and their caregivers. Twenty-four ethnic minority families with a middle school student with asthma were randomized to immediate intervention or no treatment control. Intervention students received six group sessions on prevention and management of asthma. Caregivers received five group sessions teaching child-rearing skills to support the youth's autonomy and asthma self management. All students attended all sessions; caregivers attended an average of three. Two months post-intervention, relative to controls, intervention caregivers reported better problem-solving with children. Intervention students were more responsible for carrying medication, took more prevention steps, and woke fewer nights from asthma. The intervention resulted in positive short-term changes in family relations, asthma management by students, and health status. PMID- 18411833 TI - Incorporating family therapy into asthma group intervention: a randomized waitlist-controlled trial. AB - Asthma psychoeducational programs have been found to be effective in terms of symptom-related outcome. They are mostly illness-focused, and pay minimal attention to systemic/familial factors. This study evaluated a novel asthma psychoeducation program that adopted a parallel group design and incorporated family therapy. A randomized waitlist-controlled crossover clinical trial design was adopted. Children with stable asthma and their parents were recruited from a pediatric chest clinic. Outcome measures included, for the patients: exhaled nitric oxide (eNO), spirometry, and adjustment to asthma; and for the parents: perceived efficacy in asthma management, Hospital Anxiety and Depression Scale anxiety subscale, Body Mind Spirit Well-being Inventory emotion subscale, and Short Form 12 health-related quality of life scale. Forty-six patients participated in the study. Attrition rates were 13.0% and 26.0% for the active and control groups, respectively. Repeated-measures ANOVA revealed a significant decrease in airway inflammation, as indicated by eNO levels, and an increase in patient's adjustment to asthma and parents' perceived efficacy in asthma management. Serial trend analysis revealed that most psychosocial measures continued to progress steadily after intervention. Significant improvements in both symptom-related measures and mental health and relationship measures were observed. The findings supported the value of incorporating family therapy into asthma psychoeducation programs. PMID- 18411834 TI - Asthma theory and practice: it's not too simple. PMID- 18411835 TI - Disciplinary baptisms: a comparison of the naming stories of genetics, molecular biology, genomics, and systems biology. AB - Understanding how scientific activities use naming stories to achieve disciplinary status is important not only for insight into the past, but for evaluating current claims that new disciplines are emerging. In order to gain a historical understanding of how new disciplines develop in relation to these baptismal narratives, we compare two recently formed disciplines, systems biology and genomics, with two earlier related life sciences, genetics and molecular biology. These four disciplines span the twentieth century, a period in which the processes of disciplinary demarcation fundamentally changed from those characteristic of the nineteenth century. We outline how the establishment of each discipline relies upon an interplay of factors that include paradigmatic achievements, technological innovation, and social formations. Our focus, however, is the baptism stories that give the new discipline a founding narrative and articulate core problems, general approaches and constitutive methods. The highly plastic process of achieving disciplinary identity is further marked by the openness of disciplinary definition, tension between technological possibilities and the ways in which scientific issues are conceived and approached, synthesis of reductive and integrative strategies, and complex social interactions. The importance--albeit highly variable--of naming stories in these four cases indicates the scope for future studies that focus on failed disciplines or competing names. Further attention to disciplinary histories could, we suggest, give us richer insight into scientific development. PMID- 18411836 TI - Historical ecology of the Raja Ampat Archipelago, Papua Province, Indonesia. AB - This work presents a review of the status of marine resources of the Raja Ampat Archipelago, Papua Province, Indonesia, based on narratives of early European expeditions in various museums and libraries in Europe, Canada, and local archives in Papua. More than 500 pertinent documents on the study area were identified and located in various European museums and at the University of British Columbia library. About half of these were scanned (25,000 pages), which yielded the equivalent of 900 pages of text (or 4% of the total number of pages scanned) with observations on abundance and impact of the human population on the marine ecosystem within 2 degrees North and 2 degrees South between 127 degrees and 132 degrees East. In general, these observations, which spanned the period from 1810 to the present, suggest a decrease in the perceived occurrences of turtles, fish, and invertebrates; perceived abundance of turtles, fish, and algae; percieved subsistence exploitation of marine resources; and an increase in perceived commercial exploitation of marine resources. We conclude with a discussion of the problems and potential of contents analysis, and its use in the historical reconstruction of broad biodiversity trends. PMID- 18411837 TI - Darwin's artificial selection analogy and the generic character of "phyletic" evolution. AB - This paper examines the way Charles Darwin applied his domestic breeding analogy to the practical workings of species evolution: that application, it is argued, centered on Darwin's distinction between methodical and unconscious selection. Methodical selection, which entailed pairing particular individuals for mating purposes, represented conditions of strict geographic isolation, obviously useful for species multiplication (speciation). By contrast, unconscious selection represented an open landmass with a large breeding population. Yet Darwin held that this latter scenario, which often would include multiple ecological subdistricts and thus partial isolation, was better suited for speciation than were isolated conditions. At the same time, many passages in Darwin's writings that apparently portrayedphyletic evolution exclusively (these including references to unconscious selection), actually applied to speciation as well, for phyletic change in a single district could constitute a local manifestation of a larger common-descent pattern. This generic use of "phyletic" change was reflected in Darwin's deployment of the unconscious selection analogy in his published writings as well as in his dispute with Moritz Wagner over the necessity of geographic isolation for speciation. We can thus understand Darwin's otherwise puzzling declaration in The Origin of Species that unconscious selection was 'more important' than the methodical approach. PMID- 18411838 TI - Collective phenomena and the neglect of molecules: a historical outlook on biology. PMID- 18411839 TI - An outline of the foundations of systematics and biogeography. PMID- 18411840 TI - Modelling NiFe hydrogenases: nickel-based electrocatalysts for hydrogen production. AB - NiFe hydrogenases are unique enzymes that catalyze the H+/H2 interconversion with remarkable efficiency. The determination of the tridimensional structure of their active site (a sulfur-rich dinuclear nickel-iron cluster with diatomic cyanide and carbonyl ligands) has stimulated the synthesis of a variety of nickel-based complexes as potential electrocatalysts for hydrogen production. These catalysts may provide an adequate alternative to platinum. This paper gives an historical perspective of this biomimetic structural approach and then focusses on recently reported bio-inspired functional mimics displaying electrocatalytic activity for hydrogen production. PMID- 18411841 TI - V2O5/SiO2 surface inspired, silsesquioxane-derived oxovanadium complexes and their properties. AB - Inspired by surface species proposed to occur on heterogeneous catalysts novel oxovanadium(v) silsesquioxanes were synthesised. Reaction of a T8-silsequioxane containing two geminal OH groups with O=V(O(i)Pr)3 led to a dinuclear compound where the geminal disiloxide functions of two silsesquioxanes are bridging two O=V(O(i)Pr) moieties (2). Formation of 2 shows that--in contrast to proposals made for silica surfaces--in molecular chemistry a bidentate coordination of geminal siloxides to one vanadium centre is not favourable. With the background that species being doubly anchored to a support have been suggested to play active roles on V2O5/SiO2 catalysts an anionic complex has been prepared where a divalent dioxovanadium unit replaces one Si corner of a (RSiO1.5)8, cube (a Si-OH function remains pending) (3). 3 has been intensely investigated by vibrational spectroscopy, and to support assignments not only of the v(V=O) bands but also of the v(V-O-Si) bands, whose positions are of interest in the area of heterogeneous catalysis, isotopic enrichment studies and DFT calculations have been performed. The corresponding investigations were aided by the synthesis and analysis of a silylated derivative of 3, 4. Moreover, with regard to their potential as structural and spectroscopic models all complexes were characterised by single crystal X-ray diffraction. Finally, 2 and 3 were tested as potential catalysts for the photooxidation of cyclohexane and benzene with O2. While 2 shows a slightly higher activity than vanadylacetylacetonate, 3 leads to significantly increased turnover numbers for the conversion of benzene to phenol. PMID- 18411842 TI - Bulky aryl functionalized carbazolyl ligands: amido alternatives to the 2,6 diarylphenyl ligand class? AB - Sterically encumbered amido ligands based on a 1,8-diarylcarbazol-9-yl backbone have been investigated as electronically distinct alternatives to the widely-used terphenyl ligand class in the stabilization of low-coordinate metal complexes, and structurally characterized for the first time. While 1,8-diphenylcarbazol-9 yl derivatives are readily available, facile rotation about the Cipso-Cipso bonds leads to structurally characterized main group derivatives {e.g., [(1,8-Ph,-3,6 Me2C12H4N)K]2} in which the coordination geometry at the metal centre is augmented by secondary metal arene interactions. By contrast, the extra bulk inherent in the corresponding 1,8-dimesityl ligand results in essentially perpendicular alignments of the arene and carbazole planes, and a substituent enforced sterically protected pocket. Comparative structural studies of complexes containing the GaCl2 fragment imply that the 1,8-dimesitylcarbazol-9-yl framework offers greater steric protection at the metal centre than does the corresponding 2,6-dimesitylphenyl ligand. PMID- 18411843 TI - Investigating CN- cleavage by three-coordinate M[N(R)Ar]3 complexes. AB - Three-coordinate Mo[N((t)Bu)Ar]3 binds cyanide to form the intermediate [Ar((t)Bu)N]3Mo-CN-Mo[N((t)Bu)Ar]3 but, unlike its N2 analogue which spontaneously cleaves dinitrogen, the C-N bond remains intact. DFT calculations on the model [NH2]3Mo/CN- system show that while the overall reaction is significantly exothermic, the final cleavage step is endothermic by at least 90 kJ mol(-1), accounting for why C-N bond cleavage is not observed experimentally. The situation is improved for the [H2N]3W/CN- system where the intermediate and products are closer in energy but not enough for CN- cleavage to be facile at room temperature. Additional calculations were undertaken on the mixed-metal [H2N]3Re+/CN- /W[NH2]3 and [H2N]3Re+/CN-/Ta[NH2]3 systems in which the metals ions were chosen to maximise the stability of the products on the basis of an earlier bond energy study. Although the reaction energetics for the [H2N]3Re+/CN /W[NH2]3 system are more favourable than those for the [H2N]3W/CN- system, the final C-N cleavage step is still endothermic by 32 kJ mol(-1) when symmetry constraints are relaxed. The resistance of these systems to C-N cleavage was examined by a bond decomposition analysis of [H2N]M-L1[triple bond]L2-M[NH2]3 intermediates for L1[triple bond]L2 = N2, CO and CN which showed that backbonding from the metal into the L1[triple bond]L2 pi* orbitals is significantly less for CN than for N2 or CO due to the negative charge on CN- which results in a large energy gap between the metal d(pi), and the pi* orbitals of CN-. This, combined with the very strong M-CN- interaction which stabilises the CN intermediate, makes C-N bond cleavage in these systems unfavourable even though the C[triple bond]N triple bond is not as strong as the bond in N2 or CO. PMID- 18411844 TI - Synthesis, reactivity and complexation studies of N,S exo-heterodisubstituted o carborane ligands. Carborane as a platform to produce the uncommon bidentate chelating (pyridine)N-C-C-C-s(H) motif. AB - The synthesis of N,S-heterodisubstituted 1-(2'-pyridyl)-2-SR-1,2-closo-C2B10H10 compounds (R = Et, 2; R = (i)Pr, 3) has been accomplished starting from 1-(2' pyridyl)-l,2-closo-C2B10H11 (1), and their partial deboronation reaction leading to the structurally chiral [7-(2'-pyridyl)-8-SR-7,8-nido-C2B9H10]-derivatives (R = Et, [4]-; R = (i)Pr, [5]-) has been studied. Capillary electrophoresis combined with the chiral selector alpha-cyclodextrin has permitted the separation of the electrophoretically pure racemic [7-(2'-pyridyl)-8-SR-7,8-nido-C2B9H11]- ions into two peaks each one corresponding to the interaction of one enantiomer with the alpha-cyclodextrin. The N,S-heterodisubstituted o-carborane containing a mercapto group, 1-(2'-pyridyl)-2-SH-1,2-closo-C2B10H10, 1, is one of the two examples of a rigid bidentate chelating (pyridine)N-C-C-C-S(H) motif having been structurally fully characterized. To study the potential of such a binding site, 1 has been tested as a ligand with metal ions requiring different coordination numbers, two (Au(+)) and four (Pd2+ and Rh+). The crystal structures of the Pd(II) and Au(I) complexes are reported. For the Pd(II) complex, 1 acts as a bidentate ligand whereas for Au(I), 1 acts as a monodentate ligand through the thiolate. PMID- 18411845 TI - Strong electron-accepting methylviologen dication confined in magnetic hosts: synthesis, structural characterization, charge-transfer and magnetic properties of {(MV)2[Ni(SCN)5].Cl.2H2O}n and {(MV)[m(N3)2(SCN)2]}n (M=Mn, Co). AB - Three hybrid host-guest compounds {(MV)2[Ni(SCN)5].Cl.2H2O}n (1), {(MV)[Mn(N3)2(SCN)2]}n (2) and {(MV)[Co(N3)2(SCN)2}n (3) (where MV2+ = methylviologen dication) have been obtained by self-assembly methods and characterized by X-ray crystallography, spectral methods and magnetic measurements. Compound 1 shows a quasi-two-dimensional structure which is formed by novel single thiocyanate-bridged Ni(II) chains connected through S...S interactions. Compounds 2 and 3 are isostructural, containing single micro(1,3) azide bridged Mn(II)/Co(II) square layers. MV2+, as a strong electron-acceptor and a template, is embedded between the anionic layers in all three compounds. The charge-transfer (CT) interactions between MV2+ and the anionic hosts have been revealed by structural analysis and UV-vis diffuse reflection spectroscopy. The magnetic studies of the compounds show antiferromagnetic interactions between adjacent metal ions (J = -34.52(7) cm(-1) for 1, J = -3.90(2) cm(-1) for 2 and J = -10.96(6) cm(-1) for 3). PMID- 18411846 TI - Alkali metal complexes of a naphthylamine-substituted phosphanide. AB - The reaction between {(Me3Si)2CH}PCl2 and one equivalent of [C10H6-8-NMe2]Li, followed by in situ reduction with LiAlH4, gives the secondary phosphane {(Me3Si)2CH}(C10H6-8-NMe2)PH(1) in good yield as a colourless crystalline solid. Metalation of 1 with Bu(n)Li in diethyl ether gives the lithium phosphanide [{[{(Me3Si)2CH}(C10H6-8-NMe2)P]Li}2(OEt2)](2), which undergoes metathesis with either NaOBu(t) or KOBu(t) to give the heavier alkali metal derivatives [[{(Me3Si)2CH}(C10H6-8-NMe2)P]-Na(tmeda)](3) and [[{(Me3Si)2CH}(C10H6-8 NMe2)P]K(pmdeta)](4), after recrystallisation in the presence of the corresponding amine co-ligand [tmeda = N,N,N',N'-tetramethylethylenediamine, pmdeta = N,N,N',N",N"-pentamethyldiethylenetriamine]. Compounds 2-4 have been characterised by 1H, 13C{1H} and 31P{1H} NMR spectroscopy, elemental analyses and X-ray crystallography. Dinuclear 2 crystallises with the phosphanide ligands arranged in a head-to-head fashion and is subject to dynamic exchange in toluene solution; in contrast, compounds 3 and 4 crystallise as discrete monomers which exhibit no dynamic behaviour in solution. DFT calculations on the model compound [{[(Me)(C10H6-8-NMe2)P]Li},(OMe2)] (2a) indicate that the most stable head-to head form is favoured by 15.0 kcal mol(-1) over the corresponding head-to-tail form. PMID- 18411847 TI - Expansion of iridaborane clusters by addition of monoborane. Novel metallaboranes and mechanistic detail. AB - This work reports the results of a thermally driven cluster expansion of arachno 1-{eta5-C5Me5IrH2}B3H7, 1, with BH3.THF. In addition to the previously reported product, arachno-1-{eta5-C5Me5IrH}B4H9, 2, formed at lower temperatures, reaction at 100 degrees C permits the isolation of four new iridaboranes. Two products, nido-1-(eta5-C5Me5Ir)B5H9, 3, and nido-3-(eta5-C5Me5Ir)B9H13, 4, contain a single Ir atom and five and nine framework boron atoms, respectively. One, nido-3,4 (eta5-C5Me5Ir)2B8H12, 5, contains two Ir atoms and eight framework boron atoms. Their structures are predicted by the electron counting rules to be a nido iridahexaborane, 3, nido-iridadecaborane, 4, and nido-diiridadecaborane, 5. The accuracy of these predictions in each case is established experimentally by spectroscopic characterization in solution and structure determinations in the solid state. A less stable metallaborane has been identified and the available spectroscopic and crystallographic information are consistent with the formulation nido-3,4-(eta5-C5Me5Ir)2B8H13(eta-BH2), 6, i.e., a species containing an exopolyhedral bridging BH group. These new observations, along with earlier ones on ruthenaborane cluster systems, are used to fully define a general mechanism for a cluster expansion reaction, i.e., addition of borane to form an exopolyhedral adduct followed by cage insertion. PMID- 18411848 TI - Vectorial property dependence in bis {4'-(n-pyridyl)-2,2':6',2" terpyridine}iron(II) and ruthenium(II) complexes with n = 2, 3 and 4. AB - A comparative structural and spectroscopic investigation of the complexes [M(1)2]2+, [M(2)2]2+ and [M(3)2]2+ in which M = Fe or Ru, and ligands 1, 2 and 3 are 4'-(2-pyridyl)-, 4'-(3-pyridyl)- and 4'-(4-pyridyl)-2,2':6',2"-terpyridine, respectively, is reported. The complexes [Ru(1)2]2+, [Ru(2)2]2+ and [Ru(3)2]2+ undergo mono- and bis-N-methylation. The consequences of methylation on the absorption spectra and electrochemical properties are discussed; the solid-state structure of the bis(N-methylated) derivative of [Ru(2)2][PF6]2 is presented. PMID- 18411849 TI - Efficient addition of alcohols, amines and phenol to unactivated alkenes by Au(III) or Pd(II) stabilized by CuCl2. AB - The nucleophilic addition of alcohols, amines and phenol to unactivated alkenes catalyzed by cationic gold and palladium becomes limited due to the fast reduction into metallic gold under reaction conditions. The presence of CuCl2 retards the reduction of Au(III) and Pd", strongly increasing the turnover number of gold and palladium catalysts. It is shown that new Au(III)-CuCl2 and Pd(II) CuCl2 catalysts are active and selective for the nucleophilic addition of alcohols, amines and phenol to unactivated alkenes. PMID- 18411850 TI - Molecular structures of M(Bu(t))3 (M = Al, Ga, In) using gas-phase electron diffraction and ab initio calculations: experimental and computational evidence for charge-transfer processes leading to photodissociation. AB - The gas-phase structures of AI(Bu')3 and Ga(Bu')3 have been investigated by electron diffraction and are shown to consist of monomeric units with very slightly pyramidal geometries. Salient structural parameters (r(hl)) include r(A1 C) = 2.008(2) A and r(Ga-C) = 2.032(2) A. For both compounds the ligand orientations and geometries are controlled by interligand interactions. The structures of M(Bu(t))3 (M = Al, Ga, In) have been calculated ab initio and those for the aluminium and gallium derivatives are in good agreement with the electron diffraction structures. Comparison of the ab initio calculated structure of In(Bu')3 with those of Al(Bu(t))3 and Ga(Bu(t))3 suggests that the significantly different photochemistry exhibited by the former does not result from structural factors. In fact the compounds undergo a charge-transfer process in the UV region, with the wavelength required calculated to be slightly longer for the indium compound than for the other two. PMID- 18411851 TI - The future is Bright Futures. PMID- 18411852 TI - Updates in health supervision guidelines for children, Part 1. PMID- 18411853 TI - A 17-year-old girl with fever, chills, rib, and pelvic pain. PMID- 18411854 TI - Neonatal extrapyramidal movements. Neonatal withdrawal due to maternal citalopram and ondansetron use. PMID- 18411855 TI - Poor neonatal adaptation in term infant. Fluoxetine toxicity. PMID- 18411856 TI - History of Bright Futures. PMID- 18411857 TI - Well-child care: looking back, looking forward. PMID- 18411858 TI - Bright Futures: the screening table recommendations. PMID- 18411859 TI - Practice improvement: child healthcare quality and Bright Futures. PMID- 18411860 TI - Care for children and evidence-based medicine. AB - Child healthcare sits between two worlds: in the past, with a focus on the prevention and treatment of infectious and chronic disease and the current more expansive view of child health and our role in child health promotion. As a result, disagreements concerning the evidence basis for our work have also arisen. Clinicians may stay on track by considering multiple sources of evidence in making clinical decisions. In doing so, it helps to consider each activity from the perspective of the child, family, and community. Screening and intervention may look for existing problems, for risk and resilience factors, and for opportunities to optimize each child's developmental potential. Integration of these various concerns underlies the art of modern practice, provides the assistance that families want, and leads to continued professional satisfaction. PMID- 18411861 TI - Discerning bright futures of electronic health records. AB - The recent release of the Bright Futures Guidelines, third edition, brings new opportunity and new power to our work with children and their families. Pediatricians, with their colleague nurse practitioners and family physicians, now address "new morbidities" of behavioral and psychosocial problems, developmental disabilities, and environmental stressors, in addition to the traditional morbidities of infection or malnutrition. Properly designed EHRs can add efficiency to this important work. Schor has discussed the changing nature of the health supervision encounter. If these well child visits are to remain relevant and vibrant, if clinicians are to effectively address the physical, emotional, and social health needs of our children and adolescents, new models of data management are essential. This is the challenge in the development of electronic health records for the primary care of children and youth. PMID- 18411862 TI - Providing well child care for military families: what every provider needs to consider. PMID- 18411863 TI - Animal-assisted therapy with children suffering from insecure attachment due to abuse and neglect: a method to lower the risk of intergenerational transmission of abuse? AB - Children suffering from insecure attachment due to severe abuse and/or neglect are often characterized by internal working models which, although perhaps adaptive within the original family situation, are inappropriate and maladaptive in other relationships and situations. Such children have a higher probability than the general population of becoming abusing or neglecting parents. Besides the usual goals of psychotherapy, an overall goal is to stop the cycle of abuse in which abused children may grow up to be abusing parents. Therapy with these children is complicated by their distrust in adults as well as difficulties in symbolization due to trauma during the preverbal stage. Animal-Assisted Therapy (AAT) provides avenues for circumventing these difficulties, as well as providing additional tools for reaching the inner world of the client. This article gives a brief background of the connection between insecure attachment and intergenerational transmission of abuse and neglect as well as a brief overview of the principles of AAT in a play therapy setting. A rationale for the use of AAT as a unique therapy technique for children having suffered from abuse and neglect is followed by a number of clinical examples illustrating AAT. PMID- 18411864 TI - Early identification of mental health needs for children in care: a therapeutic assessment programme for statutory clients of child protection. AB - Infants, children and young people enter out of home care in distressing circumstances. They have often suffered traumatic experiences and have had their primary relationships disrupted. They and their families and carers have specific mental health needs at this time but few have attended a mental health service. A systematic comprehensive therapeutic assessment approach is described for all child protection clients who entered out-of-home care for the first time in one calendar year. The work of the programme is described using both case study and assessment findings. More than 60 per cent of participants met criteria for a major psychiatric diagnosis, with Post Traumatic Stress and Adjustment Disorders being the most common diagnostic categories. Nearly three-quarters of participants over the age of 5 years scored in the borderline or abnormal range on the Strengths and Difficulties Questionnaire (SDQ; Goodman, 1997). There was an over-representation of infants less than 1 year of age and adolescents in the cohort, representing particular developmental challenges. The findings are discussed within the context of specific mental health service delivery to this vulnerable population of maltreated infants, children and young people. PMID- 18411865 TI - Self- and parent assessment of mental health: disagreement on externalizing and internalizing behaviour in young refugees from the Middle East. AB - Self- and parent assessment of mental health problems yield a limited degree of cross-informant agreement in adolescent populations. Working with data from 122 refugee children, adolescents and young adults from the Middle East, the aims of this study were to analyse levels of agreement and disagreement between self- and parent ratings of externalizing and internalizing behaviour and to identify predictors for the differences between the two sets of ratings. Parents and children were interviewed separately using structured questionnaires. Mental health was assessed using the Achenbach System of Empirically-based Assessment. Self- and parent-rated scale scores correlated moderately. The mean score differences between self- and parent-rated internalizing and externalizing behaviour were 2.0 and 2.7, p < 0.005, respectively. A larger mean difference was found among boys concerning externalizing behaviour and among girls concerning internalizing behaviour. Individual (age, and sex) family (father's health situation) and ethnic background predicted this difference. This could indicate that parent ratings and children's self-ratings are two, qualitatively different constructs and not just a result of expected inter-observer disagreement. When assessing young refugees for possible treatment, this difference needs to be understood and taken into consideration. PMID- 18411866 TI - Making child mental health 'everybody's business': a training course for family support workers. AB - This article provides an overview of the development and delivery of a mental health teaching and training programme offered to Family Support Workers (FSWs) working in a range of schools in an economically deprived part of Bedfordshire. The FSWs had received no prior teaching in the area of child mental health, or in ways of engaging and communicating with children and families. Both topics were considered crucial to their role. The training was delivered in two phases. Phase 1 consisted of 4 full days of teaching. Topics covered included (1) engaging children and families, (2) child development, (3) self-awareness, and (4) systemic thinking. Phase 2 was delivered through 'practice sets', which were held monthly for 4 months, and lasted 2 hours. These were used to develop the FSWs' theoretical understanding of the issues, and link this with their practice. The training was evaluated using a semistructured questionnaire. Results suggested that participants had changed the ways in which they engaged families and children. They were more able to adopt a collaborative, nonexpert, position which focused on empowering families to make decisions for themselves. This represented a significant shift in the way they worked. This process was enhanced by their more sophisticated level of understanding of child development and behaviour problems, which took into account the impact of wider systemic and relational factors and moved away from locating problems within individuals. Participants also demonstrated the ability to reflect on how their personal beliefs and experiences impacted on the work they undertook, and why this was important. Recommendations following the training included the need for regular, ongoing practice support, the need for the training to be more widely disseminated (e.g. to school staff), and the benefits of having clear descriptions of the roles and responsibilities of Family Support Workers. PMID- 18411867 TI - Use of the ADOS and ADI-R in children with psychosis: importance of clinical judgment. AB - The Autism Diagnostic Observation Schedule (ADOS) and the Autism Diagnostic Interview - Revised (ADI-R) are considered to be the 'gold standard' in diagnostic evaluations for autism. Developed as research tools and now gaining wide clinical use, the ADOS/ADI-R assessment package has been demonstrated to differentiate children with autism from those with other developmental disabilities; however, little work concerning the reliability and validity of the tools in children with a known history of psychosis has been undertaken. We report on the administration of the ADOS, ADI-R and clinical judgment in three cases of Childhood-Onset Schizophrenia. All 3 children met both ADOS and ADI-R criteria for an autism spectrum diagnosis, even though none of them received a clinical diagnosis of autism from either a research child psychiatrist or an experienced clinically trained, research psychologist with expertise in autism. Issues concerning overlap of symptom presentation and implications for research and clinical use of these assessment tools are discussed. PMID- 18411868 TI - E-mail: a new technique for forming a therapeutic alliance with high-risk young people failing to engage with mental health services? A case study. AB - This article describes e-mail therapy with a teenager whom professionals found difficult to communicate with and understand. Anne had a 4-year history of low mood and unpredictable self-harming behaviour that in the past had led to admission to a secure unit. Therapy involved weekly e-mails over a 3-month period and ended when she was transferred to adult services. The article focuses on how e-mail enabled engagement to occur. The therapeutic relationship allowed Anne to explore and understand past experiences, and start resolving present problems. Diagnosis also became clearer. PMID- 18411870 TI - Coping, social relations, and communication: a qualitative exploratory study of children of parents with cancer. AB - The purpose of this qualitative study of families where a parent has cancer was to explore ways of informing the child of the parent's illness, how the child perceives the parent's emotional state, how the child copes with the parent's illness, and how this coping relates to the parent's coping and concerns for the child. Twenty-one children from 15 families and their parents were interviewed. In 13 families the mother was ill, in two the father. Children were aware of the facts of the illness, but there was limited emotional communication between the generations. The children were very observant of both the ill and the healthy parent's emotional condition. The children's observations and expressions led us to identify five coping strategies the younger generation used: Helping others, parentification, distraction, keeping it in the head, and wishful thinking. Both adaptive and destructive examples of parentification were found. Communication patterns and parental coping seemed to be highly related to the child's coping repertoire. Even though most children seemed to manage rather well, all children were strongly affected by the illness. The 'healthiest' adaptation related to factors within the family system, which has implications for the provision of help. PMID- 18411869 TI - HIV-positive mothers' disclosure of their serostatus to their young children: a review. AB - One challenge faced by mothers living with HIV (MLWHs) is the decision about whether or not to disclose their HIV status to their young children, and how best to carry out the disclosure. Disclosure of their serostatus has emerged as one of the main concerns MLWHs have, and that decision can result in high levels of psychological distress. Concerns are exacerbated among MLWHs with younger children, due to the fact that they face additional worries, such as whether the child is old enough to understand, or will be able to keep the information confidential. A great deal of recent research--within approximately the past decade--has been conducted to investigate maternal disclosure of HIV, and the outcomes on children. This article reviews the current state of the research literature, focusing on factors that appear to influence whether or not mothers chose to disclose; characteristics of children who have been made aware of their mothers' serostatus relative to children who remain unaware; factors that appear to influence children's reactions to maternal disclosure; and implications of this research as well as future research directions. PMID- 18411871 TI - Psychoanalytic psychotherapy assessment in a child and adolescent mental health setting: an exploratory study. AB - Undertaking assessment is a significant component of a Child Psychotherapist's work within Child and Adolescent Mental Health Services (CAMHS), yet as an activity it has been relatively neglected in the research literature. This study made use of a small-scale, qualitative design to explore the nature and function of psychoanalytic assessment work within a single CAMHS team. After an initial audit of child psychotherapy assessment work, indepth interviews with 5 Child and Adolescent Psychotherapists, exploring the nature of assessment work, were analysed using Interpretative Phenomenological Analysis (IPA). Results suggested that assessment is a major part of the Child Psychotherapist's work, although there are different types of assessment done in different contexts. Among the participants there was a certain shared understanding of the psychoanalytic approach to assessment, although with significant differences in regard to process, technique (e.g. use of interpretation, the role of countertransference) and the reporting of assessments. The analysis also suggested tensions between the role of the assessor as an 'expert' and as a 'therapist'. PMID- 18411872 TI - Mothers' explanations of communicating past health and future risks to survivors of childhood cancer. AB - Mothers of survivors of Retinoblastoma (Rb) experience unique challenges communicating with their child about the condition. Children are mostly diagnosed within their first year but the consequences continue into young adult life. Here 39 mothers of Rb survivors (23 males, mean age = 10.26 years) were interviewed about their experiences. Mothers were asked about communication with their children about Rb, and future health risks. Interviews were analysed using thematic analysis. Mothers reported that they had informed children about past diagnosis and treatment but had spoken less about genetic risk or risk of secondary cancer. The child's age and information-seeking behaviour were associated with mothers' disclosure, along with mothers' perceptions that information would facilitate child coping. Findings suggest that mothers may need more guidance during follow-up care in communicating about the disease and its consequences for future health. Medical staff should also take extra care to ensure that mothers are aware of genetic counselling services and how to access them before the child is discharged from specialist care. PMID- 18411873 TI - Service delivery in child psychiatry: provider shortage isn't the only problem. AB - Although the shortage of pediatric mental health practitioners in the United States is often cited as the major stumbling block to good care, several deficiencies and problems related to service delivery may be just as great a threat to the delivery of child and adolescent psychological and psychiatric clinical care. Problems and suggestions for change related to research, training, insurance, hospitalization, practitioner collaboration and role definition are discussed. PMID- 18411874 TI - Malpractice consult. What to do when patients complain. PMID- 18411875 TI - Selling your practice in a buyer's market. PMID- 18411876 TI - Still a privilege to be a doctor. PMID- 18411877 TI - Why we've never been sued. PMID- 18411879 TI - Best ways to communicate with specialists. PMID- 18411878 TI - He was a captain to the end. PMID- 18411880 TI - Try diet counseling the easier way. PMID- 18411881 TI - A lighthearted way to attract new patients. PMID- 18411882 TI - Choosing the right life insurance. PMID- 18411883 TI - The foolproof way to hire office staff. PMID- 18411885 TI - If your EHR web host locks you out. PMID- 18411884 TI - Fighting through the EHR jungle. PMID- 18411886 TI - Writing for publication--getting started. PMID- 18411887 TI - The association of Nursing Home Compare quality measures with market competition and occupancy rates. AB - Since 2002, the Centers for Medicare and Medicaid Services have reported quality measures on the Nursing Home Compare Web site. It has been assumed that nursing homes are able to make improvements on these measures. In this study researchers examined nursing homes to see whether they have improved their quality scores, after accounting for regression to the mean. Researchers also examined whether gains varied according to market competition or market occupancy rates. They identified some regression to the mean for the quality measure scores over time; nevertheless, they also determined that some nursing homes had indeed made small improvements in their quality measure scores. As would be predicted based on the market-driven mechanism underlying quality improvements using report cards, the greatest improvements occurred in the most competitive markets and in those with the Lowest average occupancy rates. As policies to promote more competition in Long-term care proceed, further reducing occupancy rates, further, albeit small, quality gains will likely be made in the future. PMID- 18411888 TI - The $6 million question: can process improvement ensure appropriate hospitalizations? AB - Short, unnecessary hospitalizations are the largest contributor to erroneous Medicare payment. A team of medical, nursing, case management, and coding and billing professionals used process improvement techniques to reduce inappropriate 1-day admissions among 20 hospitals with high 1-day-stay utilization. Interventions included performance feedback, root cause analyses, process redesign, monthly progress monitoring, and quarterly pattern analyses. Over a 6 month period, the unweighted average admission error rate was reduced from 39% to 21% (p < .01). An estimated 1,396 1-day stays were prevented or denied payment, resulting in an annual savings of $6 million for Medicare's Hospital Insurance Trust Fund. PMID- 18411889 TI - Providing reliable care for patients with emergent acute myocardial infarction. AB - Reducing inconsistency and variability in emergency care processes increases chances for survival. Development of consistent processes for early risk stratification of patients with acute coronary syndrome and provision of evidence based therapy will reduce variability. A reproducible system of care for patients with acute coronary syndrome was developed. Results show that the current mean time to electrocardiogram was 9 minutes. The percent of patients receiving aspirin within 24 hours of arrival was 96%. Average door-to-reperfusion time was less than 90 minutes in 55% of patients. Reperfusion was accomplished by percutaneous coronary intervention for 92% of patients. Mortality for patients experiencing an acute myocardial infarction decreased 30%, from 8.9% in fiscal year 2003 to 6.2% in fiscal year 2007. PMID- 18411890 TI - Pediatric healthcare quality: A novel approach to lifelong child health research. AB - Medical science historically focuses on biomedical research about causes and treatment of disease. McMaster Child Health Research Institute (MCHRI), in Hamilton, Ontario, Canada, takes a different approach to research about lifelong health in children with varied chronic conditions. Staff at MCHRI propose to develop research programs to help "children with complicated lives and their families" meet the challenges of growing up in the face of threats to their health and development in order to help both children and families reach their full potential. This "noncategorical approach" to health services research is timely for addressing chronic childhood illnesses and disabilities across the life span. PMID- 18411891 TI - Factors affecting electronic health record adoption in long-term care facilities. AB - Electronic health records (EHRs) hold the potential to significantly improve the quality of care in long-term care (LTC) facilities, yet limited research has been done on how facilities decide to adopt these records. This study was conducted to identify factors that hinder and facilitate EHR adoption in LTC facilities. Study participants were LTC nurses, administrators, and corporate executives. Primary barriers identified were costs, the need for training, and the culture change required to embrace technology. Primary facilitators were training programs, well defined implementation plans, government assistance with implementation costs, evidence that EHRs will improve care outcomes, and support from state regulatory agencies. These results offer a framework of action for policy makers, LTC Leaders, and researchers. PMID- 18411892 TI - Spreading improvement strategies within a large home healthcare organization. AB - This article describes the process of the spread of improvement strategies to improve relationships between professional and paraprofessional service providers within a large home healthcare organization and its partnering home health aide vendors. We describe the method for spreading the strategies, which emerged from a learning collaborative, and the successes and challenges of the spread. Two case studies highlight how the attributes of innovations and the methods for spreading them can influence the effectiveness of an improvement effort. PMID- 18411893 TI - New York recognizes Joint Commission accreditation for office-based surgery. PMID- 18411895 TI - Update: methods to transmit fire alarm signals--manual transmission permitted under certain circumstances. PMID- 18411896 TI - Preventing accidents and injuries in the MRI suite. PMID- 18411897 TI - Update: Standards Improvement Initiative. PMID- 18411898 TI - New patient flow system tracer for critical access hospitals and hospitals. PMID- 18411899 TI - 2007 legislative activity in review: the past as prologue. PMID- 18411900 TI - A look back at 2007 regulatory activities. PMID- 18411901 TI - Hospice 2007 regulatory issues. PMID- 18411902 TI - 2008 Planning Congress: strategic recommendations meet industry challenges. PMID- 18411903 TI - A salute to Aaron Brown. Interview by Val J. Halamandaris. PMID- 18411904 TI - Strategic Planning Congress: Barbara Kennelly uplifts attendees. PMID- 18411905 TI - U.S. health system reform: must include long-term care fix. PMID- 18411906 TI - Family caregiving: heavy toll on caregivers, businesses. PMID- 18411907 TI - Chronic disease in the United States: failure to contain the containable. PMID- 18411908 TI - Telehomecare is essential to long-term home care. PMID- 18411909 TI - What makes an agency a great agency to sell for? PMID- 18411910 TI - Using your Web site as a marketing tool for long-term health care. PMID- 18411911 TI - Odyssey of CR 5550 and hospice physician certification signatures. PMID- 18411912 TI - [Perioperative antibiotic prophylaxis in obstetrics and gynecology]. AB - This article reviews current opinions originating from Evidence Based Medicine regarding perioperative antibiotic prophylaxis in obstetrics and gynecology. The appropriate use of antibiotic prophylaxis constitutes one of the most effective preventive tools available for the obstetricians and gynecologists. Lack of antibiotic prophylaxis or their improper use may result in serious iatrogenic harm with potential medical but also economic and legal consequences. PMID- 18411913 TI - [Biochemical aspects of modified, transdermal replacement hormone therapy]. AB - OBJECTIVES: The aim of the study is to evaluate the influence of modified, transdermal, hormone replacement therapy [HRT] and hormone supplement therapy [HST] on concentration of FSH , LH, E1, E2, PR, IGF1, GH and bone density of lumbar vertebrae in women with osteopenia in postmenopausal period. MATERIAL AND METHODS: 65 women were enrolled, aged from 43 to 58, divided into three groups based on a randomized list. Group I control used transdermal placebo, group II used transdermal hormonal therapy [HRT] and group III used hormonal therapy per oss [HST]. The concentrations of hormones were estimate by radioimmunoenzymatic methods. Statistic analysis was based on Statistical PL. RESULT: After transdermal hormonal therapy [HRT], the concentration of hormones is normalized and there is a significant increase of bone density of lumbar vertebrae. Hormonal supplement therapy per oss [HST]. There is the decrease of IGF1, BMD, and increase of concentrations of estrogens, prolactin, GH. CONCLUSION: 1. Transdermal, hormone replacement therapy [HRT] modulates concentration of sex hormones and bone metabolism. 2. Hyperestrogenism, hyperprolactinemia, decreased IGF1 concentration and low mineral density in women after HST may cause disorders of chemical estrogens metabolism. PMID- 18411914 TI - [Polymorhism of CYP17 gene and age of menarche]. AB - OBJECTIVES: It is believed that the age of menarche may be related to polymorphisms in the genes involved in estrogen biosynthesis. A candidate gene is CYP 17 encoding cytochrome P450 c17alpha, which mediates activity of steroid 17 hydroxylase and 17,20-lyase. A single base-pair change (T-->C) in the promoter region of CYP 17 creates a polymorphic variant with an additional SP-1 transcription factor binding site. The aim of our study was to evaluate the relationship between CYP 17 gene polymorphism and the age of menarche. MATERIALS AND METHODS: The study group was composed of 124 healthy Caucasian girls inhabiting the Lublin region (Poland). The observation had began from prepubertal period, thus allowing for the age of menarche to be recorded precisely. CYP 17 polymorphism was assessed with RLFP-PCR using Msp Al restriction endonuclease. An allele with the respective restriction site was designated as A2, whereas one resistant to restriction enzyme as A1. RESULTS: The following frequency of CYP 17 genotypes has been observed: A1/A1-36 (29.0%), A1/A2-67 (54.0%) and A2/A2-21 (17.0%). The mean age of menarche for these three genotypes did not differ significantly and was: A1/A1-13.14+/-0.92, A1/A2-12.93+/-1.04 and A2/A2-13.40+/ 0.91 years, although later age of first menstrual period in carriers of A2/A2 genotype compared with carriers of A1/A2 was nearly statistically significant (p=0.058). The distribution of CYP 17 genotypes in groups of girls with onset of menarche before and after thirteenth year of age did not differ significantly. CONCLUSION: The age of menarche in population of Polish girls is not associated with the polymorphism of CYP 17 gene. PMID- 18411916 TI - [Assessment of expression of luteinizing hormone (LH)/human chorionic gonadotropin (hCG) receptor (LH/hCGR) and hCG protein in ovarian cancer tissues]. AB - OBJECTIVES: Ovarian cancers still remain a significant worldwide epidemiological problem. The vast majority of newly diagnosed cases are in advanced clinical stages, and the five-year survival rate in all clinical stages amounts up to less than fifty percent. A better understanding of the biology of ovarian cancer can bring us closer to successful treatment and recovery. The aim of this study was to evaluate the expression of luteinizing hormone (LH)/human chorionic gonadotropin (hCG) receptor (LH/hCGR) and the expression of hCG protein in the ovarian cancer tissue. MATERIALS: Histologically proven primary ovarian cancer samples (n=20) were obtained from women undergoing gynecologic surgery. Frozen sections of ovarian cancer tissues were evaluated by means of indirect immunofluorescence. RESULTS: The expression of hCG protein was ubiquitous throughout all samples, while LH/hCGR was detected in only sixty percent of samples. Co-expression of LH/hCGR and hCG protein was detected in some individual cells in tumor tissue. Some cancer cells expressed only hCG protein, but not LH/hCGR. CONCLUSIONS: Expression of LH/hCG receptor is a characteristic feature of various histological types of ovarian cancer. Co-expression of LH/hCGR and hCG protein in individual cancer cells may point to an autocrine or paracrine mechanism of _hCG activity in the tumor microenvironment. Further studies are needed to evaluate LH/hCGR and hCG protein function in the course of neoplastic development. PMID- 18411915 TI - [Comparative efficacy and tolerability of drotaverine 80 mg and ibuprofen 400 mg in patients with primary dysmenorrhoea--protocol DOROTA]. AB - OBJECTIVE: To compare efficacy and tolerability of drotaverine 80 mg and ibuprofen 400 mg in patients with primary dysmenorrhoea. MATERIALS AND METHODS: Nine-month, multicenter (11 study centers in Poland), randomised, double-blind, phase III study, conducted in two parallel group, included 345 women between the ages of 18 and 35, suffering from primary dysmenorrhoea with presence of moderate to severe pain in each of the last 3 cycles. Patients had regular menstrual cycles and used an adequate barrier contraception method with a negative pregnancy test before randomization. Patients were given 80 mg drotaverine or 400 mg ibuprofen and asked to assess the pain intensity rated on a 4 point categorical scale (0--none, 1--mild, 2--moderate, 3--severe) at baseline and 0.5, 1, 2, 3, 4, 5 and 6 hours after the first intake and evaluate the efficacy (excellent, good, fair, poor) at the end of the treatment period. They were also asked to asses the tolerance of the medication (excellent, good, fair, poor). RESULTS: The weighted sum of pain intensity differences over the 6-hour observation period (SPID-6) was calculated. The pain intensity before the first intake was comparable in the two groups. At the time-points, the decrease of pain was always greater in the ibuprofen 400 mg group, than in the drotaverine 80 mg group and was maximum at the 4th hour in the drotaverine 80 mg group (-1.2+/ 1.18) and at the 3rd hour in the ibuprofen 400 mg group (-1.7+/-0.99). 41.8% patients treated with drotaverine and 68.6% patients treated with ibuprofen assessed drug efficacy as excellent or good. Although the patients' global assessment of tolerability was significantly better (p=0.02) with ibuprofen 400 mg (excellent or good: 86.8%), than with drotaverine 80 mg (excellent or good: 78.4%), no relevant difference between the both groups was observed for frequency, nature intensity and causality of the reported adverse events. CONCLUSIONS: After analizing the data statistically and clinically, significant differences in favour of ibuprofen 400 mg were evidenced for efficacy criteria. Ibuprofen was more efficient and the effect was faster, than drotaverine in relieving pain in dysmenorrhoea. Both drugs were well tolerated. The reported adverse events were typical drug reactions or expected events in patients suffering from dysmenorrhoea. PMID- 18411917 TI - [Laparoscopic dissection of uterine artery and coagulation utero-ovarian ligament for the treatment of symptomatic myomas]. AB - OBJECTIVES: Our purpose was to evaluate the effects and safety of laparoscopic dissection of the uterine artery and coagulation of the utero-ovarian ligament in treating symptomatic myomas. MATERIAL AND METHODS: We studied 40 women, aged 31 to 50, with symptomatic uterine fibroids undergoing laparoscopic dissection of the uterine artery and coagulation of the utero-ovarian ligament Ultrasound examination of uterus and dominant fibroid were performed. Their volume reduction was measured. Clinical response was evaluated according to questionnaire assessing the level of menstrual bleeding, pain and urgency. RESULTS: There were no complications during operations. Within 6 months after the surgery the mean uterus volume was reduced by 22% and mean volume of dominant fibroid was reduced by 51%. Six months after the surgery menstrual bleeding was reduced in case of 34/38 patients (85%), completed pain relief has been observed in case of 19/25 patients (76%). In case of 11/15 (73%) patients, a regression of urgency has been observed. CONCLUSIONS: Laparoscopic dissection of the uterine artery and coagulation of the utero-ovarian ligament is a safe and effective method of treating symptomatic myomas. It is an alternative to hysterectomy, especially for women who wish to preserve their uterus. PMID- 18411918 TI - [Rapid-FISH--fast and reliable method of detecting common numerical chromosomal aberrations in prenatal diagnosis]. AB - OBJECTIVE: In recent years, new possibilities of prenatal diagnosis have opened up, due to the development of techniques which guarantee shorter time of obtaining results. One of those methods, called Rapid-FISH (rapid fluorescence in situ hybridization), for detecting numerical aberrations of chromosomes 13, 18, 21, X and Y without culturing, enables to have the results in 2-5 days. The time necessary to obtain fetal karyotype result with the usage of the classical cytogenetic methods is about 2-3 weeks and depends mainly on the culture growth rate. DESIGN: The aim of the study was to evaluate the effectiveness of the Rapid FISH technique in detecting numerical chromosome aberrations of 13, 21, 18, X and Y in amniocytes' nuclei from amniotic fluid. MATERIALS AND METHODS: Rapid-FISH and cytogenetic analysis has been performed for 161 pregnancies in the Department of Genetics at Wroclaw Medical University during years 2005 and 2006. The FISH was performed using AneuVysion kit (Vysis), according to a standard protocol. RESULTS: All normal and abnormal results were confirmed by classical cytogenetic method (GTG banding and karyotyping). Additional chromosomal aberrations, not possible to be detected in FISH, were observed in case of two patients with normal results from FISH analysis. CONCLUSIONS: Rapid-FISH is a reliable and fast method for detecting numerical chromosomal aberrations in prenatal diagnosis and should be implemented as a routine diagnostic procedure in pregnancies with high risk of fetal aneuploidy (of chromosomes 13, 18, 21, X i Y). PMID- 18411919 TI - [Markers of oxidative stress in pregnancies complicated by pregnancy induced hypertension and intrahepatic cholestasis]. AB - OBJECTIVE: The aim of the study was to investigate levels of superoxide dismutase (CuZnSOD-1), catalase (CAT), glutathione peroxidase (GPx) and malonodialdehyde (MDA) in groups of pregnant women with pregnancy induced hypertension (PIH), hypertension recognized before pregnancy (HA) or intrahepatic cholestasis. MATERIAL AND METHODS: 33 women with PIH, 6 with HA and 12 with cholestasis were compared with 33 healthy pregnant women. Levels of enzymes were assessed in blood samples. Methods of delivery and obstetric results were presented. RESULTS: SOD and GPx levels did not differ significantly in any of the investigated groups. A tendency for lower mean levels of CAT in the group of PIH women, and a higher level of MDA in the group of women with HA has been noted. The mean CAT level was significantly lower in PIH and HA patients delivered instantaneously due to the risk of eclampsia. All mean levels of enzymes in the group of women with cholestasis were similar to the ones in the group of healthy women. Patients with PIH and HA gave birth more often by cesarean section, but the overall condition of the newborns was satisfactory. CONCLUSIONS: There is no substantial evidence that the level of oxidative enzymes in a blood sample can be an indicator of oxidative stress in pregnant women with PIH, HA or cholestasis. Although CAT levels were lower in PIH and HA women who had cesarean section due to the risk of eclampsia, there was no correlation between these enzyme levels and the clinical outcome of patients or the condition of the newborns. PMID- 18411920 TI - [Nucleated red blood cells as an early marker of intraventricular haemorrhage in newborn infants]. AB - INTRODUCTION: The aim of the paper was to evaluate the usability of nucleated blood cells count in newborn infants in the first 12 hours after their birth as an early marker of intraventricular haemorrhage. MATERIAL AND METHODS: The retrospective research included 210 newborns (both born at term??? and prematurely), delivered at the Department of Clinical Sciences of Perinatology of the I Chair of Gynecology and Obstetrics of the Medical University in Lodz, Poland from 1 January 2003 to 31 December 2006. During the first 12 hours after the birth, the abovementioned infants were diagnosed with nucleated blood cells in their blood count. The correlation between the number of nucleated blood cells and the occurrence of intraventricular haemorrhage has been investigated. Intraventicular haemorrhage was diagnosed with the use of transfontanel ultrasonography between the third and the seventh day of life. The increased number of erythroblasts was established to be on the level above 20 nucleated blood cells/100 white blood. The material was subject to statistical analysis which included the use of a Spearman's rank correlation coefficient, a chi-square test, and a Mann-Whitney test. The statistically significant level adopted equaled p<0.05. RESULTS: Out of 210 newborns included in the survey, among 71 (33.81%) intraventricular haemorrhage were diagnosed in the first week of their life. The I and II grade haemorrhage occurred in 61 newborns, which equals 85.92%, and the III or IV grade haemorrhage occurred in 10 patients (14.08%). In case of 50 newborns with IVH (70.42%) the number of nucleated blood cells exceeded 20/100 white blood cells. The average level of erythroblast for newborns with IVH equaled 87.46 (146.49), and in the group without IVH 38.86 (7.82). In order to examine the relation between the number of nucleated blood cells and the intraventricular haemorrhage a Spearman's rank correlation coefficient was used, as well as a chi-square test and a Mann-Whitney test On their basis it was concluded that there is a statistically significant relation between the level of erythroblast and the occurrence of intraventricular haemorrhage p=0.0048. CONCLUSIONS: The evaluation of the number of nucleated red blood cells in newborn infants can be an early, noninvasive marker of the intraventricular haemorrhage in newborns. PMID- 18411921 TI - [The latent infection of human papilloma virus in pregnat woman and colonization of placenta--preliminary report]. AB - Human papilloma virus infection may have a latent form without characteristic changes in Pap-smears. Some data suggests there exists a possibility of materno foetal transmission of the HPV infection. HPV infection may be one of the possible reasons for IUGR. AIM: The main aim of the study was to find DNA HPV in the Pap-smear and placentas in pregnancy complicated by the intrauterine growth restriction. MATERIAL AND METHODS: In two groups of women with normal Pap-smears, the material for DNA presence was taken from the uterine cervix and from the central part of placenta after the delivery. The study group consisted of pregnant women with the pregnancy complicated by fetal growth restriction. The control group consisted of women with normal fetal weight pregnancy. In cervical smears and placental fragments the presence of HPV DNA and typing of HPV using the PCR method has been done. RESULTS: In the control group the presence of low risk types of HPV was found but DNA HPV wasn't present in the placental fragments. In the study group, in 4 cases high risk HPV DNA was found in cervical smears and in 3 of those cases HPV DNA was also present in the placental fragments. In two cases it was type 16, in one--type 18. CONCLUSIONS: The latent form of high risk HPV infection might be the reason for materno-foetal transmission of HPV. The high rate of high oncogenic risk types of HPV in the group of pregnancies complicated by IUGR might suggest the correlation between HPV infection and IUGR etiology. PMID- 18411922 TI - [Future of prenatal cytogenetic studies: rapid aneuploidy testing or full karyotype]. AB - Thrombophilia is a congenital or acquired disorder of haemostatic imbalance leading to clot formation. Congenital thrombophilia is a result of different genetic polymorphisms in the genes coding for particular elements in coagulation and fibrinolysis processes and is connected with excessive readiness to thrombosis in the carriers the mutated alleles. A higher coagulation activity has been observed in case of pregnant women who are carriers of congenital thrombophilia, when compared to the pre-pregnancy activity. These changes concern first of all utero-placental circulation, and may lead to many complications during pregnancy such as: recurrent miscarriages, intrauterine fetal death in second and third trimester, preeclampsia/eclampsia, intrauterine growth restriction and placental abruption. Numerous research indicates that anticoagulation prophylaxis in pregnant women with the abovementioned complications in medical history might prevent a similar condition in the following pregnancies. What is more, it underlines that administration of low molecular weight heparin and acetylsalicylic acid may improve perinatal outcome in thrombophilic women. However, the notion whether anticoagulant prophylaxis should be applied in women with preeclampsia, fetal hypotrophy or fetal loss remains disputable. Furthermore, the question of when the prophylaxis should start and of its duration remains unanswered. The following summary focuses on congenital thrombophilia in pregnant women with burdened anamnesis and suggested pattern of anticoagulation prophylaxis. PMID- 18411923 TI - [DNA adducts in human female genital organs]. AB - DNA adducts, one of genetic damages markers, precede and finally can lead to oncogenic mutations. They appear in genome as a result of DNA bases damages caused by various and numerous environmental factors eg. ultraviolet light, ionic radiation, toxins and also endogenic substances, for example estrogens. It is believed that the creation of DNA adducts is a necessary but insufficient process for the neoplastic transformation of the cell. The following review presents concise knowledge about the DNA adducts creation and their sequels served in healthy and cancerous tissues of the female genital organs, on the base of the available data. PMID- 18411924 TI - [Pathogenesis and genetics of neural tube defects]. AB - After congenital heart defects neural tube defects (NTDs) is the second most prevalent congenital malformation among birth defects. The average rate of isolated NTDs is 1.4-2.0 per 1000 live births worldwide. The etiology of isolated (nonsyndromic) NTDs is believed to be the result of a combination of genetic predisposition and environmental factors. Over 80 genes believed to be engaged in the neurulations have been identified during the investigation and research of the mouse models. Despite exhaustive research efforts, now spanning several decades, little is known about the actual genetic mechanisms governing the primary events involved in neural tube closure (NTC). PMID- 18411925 TI - [New views on hyperplastic endometrial lesions classification--endometrial intraepithelial neoplasia (EIN)]. AB - The aim of the study was to present a new EIN classification of premalignant endometrial lesions. The diagnosis of precancerous disease of the endometrium remains non-standardized because the most widely used World Health Organisation classification is a poorly reproducible system, which does not specify objective architectural criteria for each category of hyperplasia and does not correspond to an appropriate clinical management (undertreatment, overtreatment of the lesions). The new proposed EIN diagnostic schema, based on integrated morphological, genetic molecular, objective histomorphometric (D-score) and clinical outcome studies, divides endometrial lesions into three categories: benign hyperplasia, endometrial intraepithelial neoplasia, and cancer. PMID- 18411926 TI - [How to treat anorexia nervosa?--case report]. AB - Anorexia nervosa is an eating disorder characterized by low body weight and body image distortion with obsessive fear of gaining weight. It is a complex condition involving psychological, neuroendocrine, hormonal and metabolic components. The following is a case report of a young patient diagnosed as anorexia nervosa and primary amenorrhea, without the development of secondary female characteristics. Hormonal examination confirmed hypothalamic origin of the described disorders. There was an important impairment in the mood sphere (Beck's Depression Inventory and Hamilton Depression Rating Scale) of this young woman. Densitometry of lumbar spine revealed significant bone loss (diagnozed as osteoporosis) in this patient. All of the abovementioned examinations were repeated after one and a half years of hormonal treatment (estrogen-progestins). A return of normal menstrual cycle (with ovulation function) and normal hormonal profile have been observed. After one and a half years of treatment the patient had regular feeding patterns and none of the anorexia nervosa symptons have been found. A significant improvement in Beck's Depression Inventory and Hamilton Depression Rating Scale was found after one and a half years of hormonal treatment. The patient was also characterized by important increase of bone mass density--osteopenia was described by the densitometry of the lumbar spine. PMID- 18411927 TI - [Pelvic actynomycosis as the result of a long standing use of an intrauterine device. Case report]. AB - The aim of the study was to introduce a case of a 49-year-old woman who had been using an intrauterine contraceptive device for 20 years. The pelvic actynomycosis in her case led to sepsis and consequently to her death. Colonisation Actinomyces israeli in women using IUD may amount up to even 20%, however infections occur very seldom. Following the recommended time of IUD usage, cytology and early start of antibiotic therapy may protect patients against the Actinomyces infection. PMID- 18411928 TI - [Central Coordinating Center. Summary of Cervical Cancer Population Screening Program in the period from January the 1st 2007 till September 30th 2007]. PMID- 18411929 TI - Highly pathogenic avian influenza: an update. PMID- 18411930 TI - Disease intelligence for highly pathogenic avian influenza. AB - A comprehensive approach to highly pathogenic avian influenza (HPAI) is crucial for identifying all the factors that contribute to its emergence, spread and persistence. Epidemiological understanding makes it possible to predict the evolution of the virus and to prevent and control the socioeconomic, environmental, institutional and policy consequences. At FAO, risk assessment and intelligence with regard to HPAI are based on lessons learnt from assisting countries to design strategies and on implementation of technical assistance programmes, which reveal important elements, such as the roles of ducks, live bird markets and trade. Wild birds were found to contribute, by transporting the H5N1 virus over long distances. The contributions of different poultry farming systems and market chains in the epidemiology of HPAI are well recognized; however, the respective roles of smallholder systems and commercial farms are unclear. FAO considers that smallholders will continue to be an important factor and should be taken into account in control and prevention programmes. Changes in poultry farming are essentially driven by the private sector and market forces and could have negative consequences on the livelihoods of smallholders and on ecologically balanced production systems and agricultural biodiversity. Biosecurity can, however, be improved at the level of farms and markets. Institutional factors, such as the capacity of animal health systems to deliver control programmes, are also important, requiring strengthening and innovation in risk analysis and management. PMID- 18411931 TI - Control strategies for highly pathogenic avian influenza: a global perspective. AB - Comprehensive programmes for the prevention, detection and control of highly pathogenic avian influenza (HPAI) require a national dimension and relevant national legislation in which veterinary services can conduct surveillance, competent diagnosis and rapid response. Avian influenza was controlled and prevented by vaccination long before the current H5N1 crisis. The use of vaccine cannot be separated from other essential elements of a vaccination campaign, which include education in poultry production practices, such as hygiene, all in all out production concepts, separation of species, biosecurity (bio-exclusion to keep the disease out and biocontainment to keep the disease from spreading once suspected or detected), competence in giving the vaccine and the role of vaccination teams, post-vaccination monitoring to ensure efficacy and to detect the circulation of wild-type virus, surveillance and buffer zones in outbreak areas, and performance indicators to determine when vaccination can cease. Reporting of disease can be improved through well-structured, adequately financed veterinary services and also by fair compensation for producers who suffer financial loss. A rapid response to suspected cases of HPAI should be ensured in simulation exercises involving various sectors of the food production and marketing chain, policy-makers, official veterinary structures and other government personnel. As for other transboundary animal diseases, national approaches must be part of a regional strategy and regional networks for cooperation and information sharing, which in turn reflect global policies and international standards, such as the quality of vaccines, reporting obligations, humane interventions, cleaning and disinfection methods, restocking times, monitoring and safe trade. PMID- 18411932 TI - International standards and guidelines for vaccination of poultry against highly pathogenic avian influenza. AB - The current strain of highly pathogenic avian influenza (HPAI), H5N1, has caused an unprecedented situation, spreading over three continents, with severe economic and social consequences. The strategy of the World Organisation for Animal Health (OIE) focuses on the following key actions: early warning, early detection, rapid confirmation of suspected cases, rapid response and rapid and transparent notification. Vaccination is one means that can be used to control the virus. During the current H5N1 outbreak, the OIE received many requests from member countries for guidance in deciding whether to vaccinate and in the design of vaccination programmes. The OIE has published a general information document on vaccination against avian influenza and a document giving guidelines for decision making, including a checklist of essentials for establishing a vaccination programme. PMID- 18411933 TI - Experiences in control of avian influenza in Europe, the Russian Federation and the Middle East. AB - An unprecedented global epidemic of highly pathogenic avian influenza virus H5N1 has and continues to present enormous challenges to the international community for control in the animal reservoir. Enhanced biosecurity, good surveillance, both passive and active, supplemented by strong veterinary services, can reduce the risk for incursion and subsequent spread in free countries. Surveillance of mortality and laboratory testing among wild birds are useful early indicators of incursion of the virus into areas in which domestic poultry are not infected. Conventional control methods used widely in Europe and the Middle Eastern region involve stamping-out, zoning, quarantine, movement restrictions, enhanced surveillance and disinfection. Use of preventive vaccination is increasing in the region. In the Russian Federation, all backyard poultry considered to be at high risk for infection have been vaccinated since 2006. Several countries in the Middle East permit the use of vaccine, although rarely as part of a formal statutory programme. In the European Union, conventional approaches for control have proved effective, but both emergency and preventive vaccination could be used. Application of such programmes would have to be preceded by an evaluation of the risks for introduction and spread and might be restricted. PMID- 18411934 TI - Experience in control of avian influenza in Asia. AB - Highly pathogenic H5N1 avian influenza viruses have been circulating in Asia for over ten years, providing considerable experience on which to base appropriate long-term strategies for their control. Experience in Hong Kong SAR demonstrates that existing production and marketing practices should be changed and a range of parallel measures used. It also shows the extent of surveillance required to ensure continuing freedom from infection. Certain high-risk practices should be changed or otherwise overcome in order to control and prevent disease, including intensive rearing of large numbers of poultry in premises without biosecurity commensurate with the level of risk for exposure; complex market chains involving many smallholders selling poultry through large numbers of transporters and middlemen in poorly regulated live poultry markets; and rearing of large numbers of ducks outdoors. These high-risk practices are compounded by weak veterinary services and poor reporting systems. In many parts of Asia, these methods of rearing and marketing are an integral way of life, support the poorest members of the community or cannot be changed quickly without severe socioeconomic consequences. The gains made so far will be ephemeral unless there is a shift from an emergency focus to one of consolidation in which these high-risk practices are identified and sustainable measures implemented to minimize the risks they pose, taking account of the socioeconomic effects of interventions. Vaccination will play a key role, as it currently does in China and Viet Nam. PMID- 18411935 TI - Experience in control of avian influenza in Africa. AB - The first outbreak of H5N1 highly pathogenic avian influenza (HPAI) in Africa was confirmed at Kaduna, Nigeria, on 8 February 2006. Within three months, seven other countries on the continent, Burkina Faso, Cameroon, Cote d'Ivoire, Djibouti, Egypt, Niger and Sudan, were infected. More recently Ghana and Togo became infected. The origin of the introduction of the disease to Nigeria and the other infected countries is still unknown, owing to lack of adequate tracing of the movements of poultry and poultry products and lack of reliable epidemiological data from the affected countries. The preventive measures adopted in countries free from H5N1 HPAI include selective or total bans on the importation of poultry and poultry products from infected countries. All the infected countries have implemented more or less the same internationally recommended disease control measures including quarantine, stamping-out and active surveillance, while poultry vaccination was carried out in Cote d'Ivoire and Egypt. These control measures, adopted and implemented by weak veterinary services, cannot explain the apparent 'epidemiological silence' of H5N1 HPAI in Africa, and further studies are needed to explain the different behaviour of the H5N1 HPAI virus in Africa and Asia. PMID- 18411936 TI - Economic issues in vaccination against highly pathogenic avian influenza in developing countries. AB - We consider the use of vaccination against highly pathogenic avian influenza (HPAI) in three contexts: as part of a stamping-out programme, as a government led action for disease prevention and as private insurance by farmers. Poultry systems in developing countries cover all four of the poultry sectors defined by FAO and the OIE, each with particular economic aspects that might motivate farmers to take part in vaccination programmes or to initiate and finance them. Outbreaks in flocks of different types have different potential impacts in terms of disease spread and economic effects, which influence the potential benefits of vaccination as a means to prevent or control outbreaks. We use data from three countries to illustrate the costs of vaccination and discuss measures of cost effectiveness and ways to improve it. We also consider the question of funding sources and their impact on the sustainability of vaccination programmes. PMID- 18411937 TI - Challenges and constraints to vaccination in developing countries. AB - The challenges and constraints to vaccinating poultry in areas where adequate infrastructure and human resources are lacking are addressed in both a technical and a socioeconomic framework. The key issues discussed are: (1) selection of an appropriate vaccine and vaccination technique, including the advantages and disadvantages of a combined vaccine against highly pathogenic avian influenza (HPAI) and Newcastle disease and addressing the differences between endemic disease and emergency disease control; (2) vaccine conservation and distribution; (3) evaluation of the flocks to be vaccinated in terms of their disease status, immunocompetence and production systems; (4) design of effective information, education and communication materials and methods with and for veterinary and extension staff as well as commercial and smallholder producers and community vaccinators in rural areas; (5) evaluation and monitoring systems for technical and socioeconomic factors that affect vaccination; (6) support and coordination of and by relevant public and private agencies; (7) the role of simultaneous implementation of other control activities in addition to vaccination; (8) the importance of assessing the costs and cost-effectiveness of various approaches to the control of HPAI, including the prevention of other endemic killer diseases and options for cost-sharing; (9) evaluation of the incentives for poultry holders, vaccinators and vaccine producers to contribute to and participate in effective vaccination campaigns; and (10) policy development and the organizational framework for short- and long-term implementation and communication to decision-makers. PMID- 18411938 TI - Needs assessment for the use of vaccination. PMID- 18411939 TI - Risk modelling for vaccination: a risk assessment perspective. AB - Any risk assessment involves a number of steps. First, the risk manager, in close liaison with the risk assessor, should identify the question of interest. Then, the hazards associated with each risk question should be identified. Only then can the risks themselves be assessed. Several questions may reasonably be asked about the risk associated with avian influenza vaccines and their use. Some apply to any vaccine, while others are specific to avian influenza. Risks may occur during manufacture and during use. Some concern the vaccines themselves, while others address the effect of failure on disease control. The hazards associated with each risk question are then identified. These may be technical errors in design, development or production, such as contamination or failure to inactivate appropriately. They may relate to the biological properties of the pathogens themselves displayed during manufacture or use, for example, reversion to virulence, shedding or not being the right strain for the subsequent challenge. Following a consideration of risks and hazards, the information needed and an outline of the steps necessary to assess the risk is summarized, for an illustrative risk question using, as an example, the risks associated with the use of vaccines in the field. A brief consideration of the differences between qualitative and quantitative risk assessments is also included, and the potential effects of uncertainty and variability on the results are discussed. PMID- 18411940 TI - Planning and executing a vaccination campaign against avian influenza. AB - Vaccination against avian influenza infection caused by H5 or H7 virus subtypes has been used on several occasions in recent years to control and in some cases eradicate the disease. In order to contain avian influenza infection effectively, immunization should be combined with a coordinated set of control and monitoring measures. The outcome of an immunization campaign depends on the territorial strategy; whereas the capacity of the veterinary services in developed countries permits enforcement of strategies aimed at eradicating avian influenza, many countries currently affected by highly pathogenic avian influenza (HPAI) H5N1 viruses have a limited veterinary infrastructure and a limited capacity to respond to such epidemics. In these countries, resources are still insufficient to conduct adequate surveillance for identification and reaction to avian influenza outbreaks when they occur. When properly applied in this scenario, immunization can reduce mortality and production losses. In the long term, immunization might also decrease the prevalence of infection to levels at which stamping-out and surveillance can be applied. Countries should adapt their immunization programmes to local conditions in order to guarantee their efficacy and sustainability. In the initial emergency phase, human resources can be mobilized, with reliance on personal responsibility and motivation, thus compensating for potential shortcomings in organization. A more appropriate allocation of resources must be pursued in the long term, remembering that biosecurity is the main component of an exit strategy and must always be improved. PMID- 18411941 TI - Current availability of vaccines including vaccine banks. AB - Avian influenza can be considered one of the greatest global challenges the animal health sector has ever had to face. It is primarily a disease of animals and must be effectively managed by the veterinary community. Infection of humans in contact with the H5N1 virus continues to occur, however, with the possibility of mutations or re-assortment and the ever-present threat of the emergence of a pandemic. Therefore, whatever measures can be taken when outbreaks of avian influenza occur to reduce the amount of virus in circulation and potential human exposure must be reviewed constantly. Conventional methods for controlling diseases of epizootic proportions, such as avian influenza, have tended to rely on bio-security and culling (stamping out) of enormous numbers of animals. Such approaches might not be adequate in areas of intensive animal husbandry, and society is questioning more and more the ethics of slaughtering millions of animals. The costs and economic impact of this strategy also has a considerable effect on the economy of the country or region in which the disease outbreak occurs. PMID- 18411942 TI - Vaccination: a means for preserving poultry genetic resources? AB - The genetic diversity of poultry worldwide is under threat. About 30% of reported poultry breeds were at risk of extinction before the crisis of highly pathogenic avian influenza (HPAI). Precise national statistics on the numbers of poultry producers, stocks, volume of production and the relative contributions of commercial and local stocks to production in developing countries are lacking. Although HPAI outbreaks have been occurring in East and South-East Asia for a number of years, many countries are still not sufficiently prepared to control outbreaks, and no contingency plans or information on spatial production include data on poultry genetic diversity. Comprehensive programmes for preventing HPAI disease should take into account the conservation of genetic diversity, which may include isolation or targeted protection of valuable stocks, through, for instance, vaccination and precautionary cryo-conservation of genetic material or other measures to ensure that genetic material can be spared from generalized culling measures in an outbreak. First, it will be necessary to characterize poultry genetic resources, and then to build inventories, including spatial information on breeds and valuable breeding stocks. Countries with low capacity to conduct vaccination campaigns often also have limited capacity to characterize or make inventories of genetic resources. Therefore, capacity- and institution building are required in the animal production and health sector to ensure effective genetic stock assessment and successful vaccination programmes to preserve poultry genetic diversity in the event of disease introduction. PMID- 18411943 TI - Current developments in avian influenza vaccines, including safety of vaccinated birds as food. AB - Until recently, most vaccines against avian influenza were based on oil emulsified inactivated low- or high-pathogenicity viruses. Now, recombinant fowl pox and avian paramyxovirus type 1 vaccines with avian influenza H5 gene inserts (+ or - N1 gene insert) are available and licensed. New technologies might overcome existing limitations to make available vaccines that can be grown in tissue culture systems for more rapid production; provide optimized protection, as a result of closer genetic relations to field viruses; allow mass administration by aerosol, in drinking-water or in ovo; and allow easier strategies for identifying infected birds within vaccinated populations (DIVA). The technologies include avian influenza viruses with partial gene deletions, avian influenza-Newcastle disease virus chimeras, vectored vaccines such as adenoviruses and Marek's disease virus, and subunit vaccines. These new methods should be licensed only after their purity, safety, efficacy and potency against avian influenza viruses have been demonstrated, and, for live vectored vaccines, restriction of viral transmission to unvaccinated birds. Use of vaccines in countries affected by highly pathogenic avian influenza will not only protect poultry but will provide additional safety for consumers. Experimental studies have shown that birds vaccinated against avian influenza have no virus in meat and minimal amounts in eggs after HPAI virus challenge, and that replication and shedding from their respiratory and alimentary tracts is greatly reduced. PMID- 18411944 TI - Efficacy of immunization in different poultry species. PMID- 18411945 TI - Diagnosing avian influenza infection in vaccinated populations by systems for differentiating infected from vaccinated animals (DIVA). AB - Vaccination against avian influenza is recommended as a tool to support control measures in countries affected by avian influenza. Vaccination is known to increase the resistance of susceptible birds to infection and also to reduce shedding; however, it does not always prevent infection. Vaccinated infected flocks can therefore be a source of infection and thus be responsible for the perpetuation of infection. To avoid the spread of infection in a vaccinated population, immunization strategies must allow differentiation of infected from vaccinated animals (DIVA), combined with an appropriate monitoring system. Vaccinated exposed flocks must be identified and managed by restriction policies that include controlled marketing and stamping-out. Several vaccines and diagnostic tests to detect infection in vaccinated populations are available, the tests having various properties and characteristics. In order to achieve eradication, the most appropriate DIVA vaccination strategy must be identified and an appropriate monitoring programme be designed, taking into account risk factors, the epidemiological situation and the socioeconomic implications of the policy. PMID- 18411946 TI - The vaccination programme in Indonesia. AB - The Indonesian response to the outbreak of highly pathogenic avian influenza (HPAI) is being strengthened by increased intersectoral commitment and greater availability of staff and resources. Vaccination against avian influenza has been used widely in large commercial sectors but less so in other sectors. Generally, there has been a reduction in outbreaks and in the impact of HPAI on the commercial industry. Afield trial is described that might provide insight into the efficacy of vaccination on farms in sector 3. Preliminary data suggest that vaccination of layers induces high titres, whereas vaccination of native chickens might be difficult owing to a low response in these breeds. A much greater commitment of management, staff and resources is required before vaccination can become part of a successful sustainable campaign to eradicate HPAI. For success, the commercial poultry industry must become an integral part of the control programme, providing information and having the opportunity to identify or modify the priorities of the control programme. PMID- 18411949 TI - [Functional recovery in elderly vascular hemiplegics]. PMID- 18411950 TI - [A night home care service, a useful or superfluous initiative?]. PMID- 18411951 TI - [From Lebanon to France, a journey for a nurse for whom life counts above all. Interview by Julie Bouchara]. PMID- 18411952 TI - [Urination and it's disorders in the elderly person]. PMID- 18411953 TI - [From urination to urinary incontinence]. PMID- 18411954 TI - [The mechanisms responsible for urinary incontinence]. PMID- 18411955 TI - [Clinical nursing management of bladder sphincter disorders]. PMID- 18411956 TI - [Nursing tools used for effective management of urinary disorders]. PMID- 18411957 TI - [Better knowing the elderly incontinent patient for optimal individualized management]. PMID- 18411958 TI - [Management of urinary incontinence, a process of care]. PMID- 18411959 TI - [2/6 The consultant nurse in nursing homes]. PMID- 18411960 TI - [The Chart of Rights and Liberties for the Elderly Dependent Person. 6/7 Care of the person at the end of life]. PMID- 18411961 TI - [Enteral feeding]. PMID- 18411962 TI - [2/6--Hygiene and maintenance of removable dental prostheses]. PMID- 18411963 TI - Fledgling firms offer hope on health costs. AB - Julia Adler-Milstein and Ashish Jha Regional health information organizations- RHIOs--are springing up in the United States to meet a vital need: to connect the nation's disparate patient-health information systems. If RHIOs can find a viable business model, they stand to improve the quality and decrease the cost of U.S. health care dramatically. PMID- 18411964 TI - Authenticity: is it real or is it marketing? AB - Marty Echt, the new head of marketing at Hunsk Engines, is determined to bring the motorcycle maker back to its roots. He says it's not enough to project authenticity to customers--employees must personally subscribe to the brand's values. Should the company's CEO support Marty's "real deal" vision? Five experts comment on this fictional case study. Bruce Weindruch, the founder and CEO of the History Factory, says that an authenticity-based campaign can be effective--but only if it's truly drawn from history. Marketers like Marty often remember their organization's past in a golden haze. Weindruch recommends exploring old engineering drawings, ads, and product photos in order to understand what customers and employees really valued back in the day. Gillian Arnold, a consultant to luxury fashion and fine jewelry brands, thinks Marty's approach is right: People in key marketing posts must be passionate about their products and know them inside and out. She argues that the CEO needs to commit more fully to the new campaign and address the significant gap between the staff and the brand. James H. Gilmore and B. Joseph Pine II, the cofounders of Strategic Horizons, point out that Hunsk needs to manage customers' perceptions rather than trying to be a "real company" or forming a management team whose personal interests match the brand. People purchase a product if it conforms to their self-image; that alone determines the brand's authenticity. Glenn Brackett of Sweetgrass Rods, a maker of bamboo fly-fishing rods, says Marty seems to be one of the few people who understand Hunsk motorcycles. If employees bring blood, sweat, heart, and soul to a product, it will manifest that spirit, and customers will line up for it. PMID- 18411965 TI - Timeless leadership. AB - The historian David McCullough, a two-time Pulitzer Prize winner and well-known public television host, has spent his career thinking about the qualities that make a leader great. His books, including Truman, John Adams, and 1776, illustrate his conviction that even in America's darkest moments the old fashioned virtues of optimism, hard work, and strength of character endure. In this edited conversation with HBR senior editor Bronwyn Fryer, McCullough analyzes the strengths of American leaders past and present. Of Harry Truman he says, "He wasn't afraid to have people around him who were more accomplished than he, and that's one reason why he had the best cabinet of any president since George Washington....He knew who he was." George Washington--"a natural born leader and a man of absolute integrity"--was unusually skilled at spotting talent. Washington Roebling, who built the Brooklyn Bridge, led by example: He never asked his people to do anything he wouldn't do himself, no matter how dangerous. Franklin Roosevelt had the power of persuasion in abundance. If McCullough were teaching a business school leadership course, he says, he would emphasize the importance of listening--of asking good questions but also noticing what people don't say; he would warn against "the insidious disease of greed"; he would encourage an ambition to excel; and he would urge young MBAs to have a sense that their work maters and to make their good conduct a standard for others. PMID- 18411966 TI - Talent management for the twenty-first century. AB - Most firms have no formal programs for anticipating and fulfilling talent needs, relying on an increasingly expensive pool of outside candidates that has been shrinking since it was created from the white-collar layoffs of the 1980s. But the advice these companies are getting to solve the problem--institute large scale internal development programs--is equally ineffective. Internal development was the norm back in the 1950s, and every management-development practice that seems novel today was routine in those years--from executive coaching to 360 degree feedback to job rotation to high-potential programs. However, the stable business environment and captive talent pipelines in which such practices were born no longer exist. It's time for a fundamentally new approach to talent management. Fortunately, companies already have such a model, one that has been well honed over decades to anticipate and meet demand in uncertain environments: supply chain management. Cappelli, a professor at the Wharton School, focuses on four practices in particular. First, companies should balance make-versus-buy decisions by using internal development programs to produce most--but not all--of the needed talent, filling in with outside hiring. Second, firms can reduce the risks in forecasting the demand for talent by sending smaller batches of candidates through more modularized training systems in much the same way manufacturers now employ components in just-in-time production lines. Third, companies can improve their returns on investment in development efforts by adopting novel cost-sharing programs. Fourth, they should seek to protect their investments by generating internal opportunities to encourage newly trained managers to stick with the firm. Taken together, these principles form the foundation for a new paradigm in talent management: a talent-on-demand system. PMID- 18411967 TI - A more rational approach to new-product development. AB - Companies often treat new-product development as a monolithic process, but it can be more rationally divided into two parts: an early stage that focuses on evaluating prospects and eliminating bad bets, and a late stage that maximizes the remaining candidates' market potential. Recognizing the value of this approach, Eli Lilly designed and piloted Chorus, an autonomous unit dedicated solely to the early stage. This article demonstrates how segmenting development in this way can speed it up and make it more cost-effective. Two classes of decision-making errors can impede NPD, the authors say. First, managers often ignore evidence challenging their assumptions that projects will succeed. As a result, many projects go forward despite multiple red flags; some even reach the market, only to fail dramatically after their introduction. Second, companies sometimes terminate projects prematurely because people fail to conduct the right experiments to reveal products' potential. Most companies promote both kinds of errors by focusing disproportionately on late-stage development; they lack the early, truth-seeking functions that would head such errors off. In segmented NPD, however, the early-stage organization maintains loyalty to the experiment rather than the product, whereas the late-stage organization pursues commercial success. Chorus has significantly improved NPD efficiency and productivity at Lilly. Although the unit absorbs just one-tenth of Lilly's investment in early-stage development, it delivers a substantially greater fraction of the molecules slated for late Phase II trials--at almost twice the speed and less than a third of the cost of the standard process, sometimes shaving as much as two years off the usual development time. PMID- 18411968 TI - Is yours a learning organization? AB - An organization with a strong learning culture faces the unpredictable deftly. However, a concrete method for understanding precisely how an institution learns and for identifying specific steps to help it learn better has remained elusive. A new survey instrument from professors Garvin and Edmondson of Harvard Business School and assistant professor Gino of Carnegie Mellon University allows you to ground your efforts in becoming a learning organization. The tool's conceptual foundation is what the authors call the three building blocks of a learning organization. The first, a supportive learning environment, comprises psychological safety, appreciation of differences, openness to new ideas, and time for reflection. The second, concrete learning processes and practices, includes experimentation, information collection and analysis, and education and training. These two complementary elements are fortified by the final building block: leadership that reinforces learning. The survey instrument enables a granular examination of all these particulars, scores each of them, and provides a framework for detailed, comparative analysis. You can make comparisons within and among your institution's functional areas, between your organization and others, and against benchmarks that the authors have derived from their surveys of hundreds of executives in many industries. After discussing how to use their tool, the authors share the insights they acquired as they developed it. Above all, they emphasize the importance of dialogue and diagnosis as you nurture your company and its processes with the aim of becoming a learning organization. The authors' goal--and the purpose of their tool--is to help you paint an honest picture of your firm's learning culture and of the leaders who set its tone. PMID- 18411969 TI - Save the children. PMID- 18411970 TI - Documentation and proper office protocol: avoiding allegations of pregnancy discrimination. PMID- 18411971 TI - And now for something completely different... PMID- 18411972 TI - New insights into ischemic heart disease in women. AB - Coronary artery disease is different in women than in men in its pathogenesis, symptoms, and prognosis. Needed is a strategy for detecting and assessing coronary disease specifically in women. This review highlights recent evidence on sex differences in coronary artery disease. PMID- 18411973 TI - Addiction in older women: American health care's best-kept secret. AB - Older women are not immune from alcohol and drug dependence. In fact, substance use disorders are becoming more common in women over 60 and will become a larger public health issue as the baby boomers reach retirement age. Addicted women in their senior years present special challenges in identification and intervention, and have special needs in treatment. Incidence of co-occurring medical and psychiatric problems is increased. They respond best to a compassionate, nonjudgmental counseling approach. PMID- 18411974 TI - Coming from all directions: protecting girls and women from the impact of substance use. AB - Dentists may assist in protecting their female (and male) patients from unhealthy substance use conditions. Prevalence is so high that daily, nearly every U.S. health care provider sees patients either at risk themselves or experiencing negative effects of substance use by a friend, family member, or co-worker. Health care practice-based interventions employ simple universal standardized screening, brief advice, and occasional referrals to specialists and/or community resources. Youth prevention strategies reduce risk and build protective factors. PMID- 18411975 TI - Musculoskeletal health of the woman dentist: distinctive interventions for a growing population. AB - Female dentists face unique musculoskeletal demands and inherent gender differences that may place them at higher risk for occupational pain and injury than their male counterparts. By familiarizing themselves with unique muscle imbalances, specific pain syndromes, and positioning challenges to which they are predisposed, female dentists may take pre-emptive action and initiate ergonomic intervention in the operatory and specific exercise at home to reduce the risk of developing musculoskeletal disorders. PMID- 18411976 TI - A whiter shade of pale. PMID- 18411977 TI - More prevention than cure. PMID- 18411978 TI - Exorcising their demons. PMID- 18411979 TI - Scotland may no longer remain 'the free' for those over 65. PMID- 18411980 TI - Setting a good example. PMID- 18411981 TI - The global perspective. PMID- 18411982 TI - Mind the gap. PMID- 18411983 TI - Brushing up on oral hygiene. PMID- 18411984 TI - How to promote a healthier tomorrow. PMID- 18411985 TI - The MCA and end-of-life care. PMID- 18411986 TI - Providing rehabilitation services for people with dementia through intermediate care. AB - This article describes a Department of Health-funded study that mapped rehabilitation services--and examples of good practice--for older people with dementia in England. The findings suggest that there are potential benefits in locating rehabilitation services within intermediate care, including a higher service profile with clearer care pathways and access to a wider range of professionals across health and social care. The author concludes that a seamless range of services can be provided via intermediate care, but argues that nurses need to challenge the perception that rehabilitation cannot be achieved for people with dementia. PMID- 18411987 TI - End-of-life care for patients with dementia. AB - Studies have shown that people with dementia receive sub-optimal end-of-life care (McCarthy et al, 1997; Morrison and Siu 2000; Mitchell et al, 2004). The National Institute of Clinical Excellence (NICE) and the National Council for Palliative Care (NCPC) have highlighted the importance of palliative care for people with dementia (NICE 2006; NCPC 2007). This article explores the palliative care needs of older people with dementia and considers how they may be addressed. PMID- 18411988 TI - Carer-driven dementia education for professionals. AB - The pilot project described here took the form of action research, focusing on dementia awareness education for nurses on acute wards and in accident and emergency departments. An essential feature of the educational package was the involvement of four carers, each of whom described specific experiences. Their accounts were recorded on DVD and presented in the education sessions, which the carers also attended to add comments and answer questions. Feedback from the nurses was positive with all of them claiming new awareness and understanding of patient and carer needs. The education sessions are to be continued with other nurses. PMID- 18411989 TI - Can a new pension scheme prevent staff shortages? PMID- 18411990 TI - 'We have a generation of nurses who do not perform many aspects of basic care'. PMID- 18411991 TI - Why are there so few men in nursing? PMID- 18411992 TI - Nursing in the 1970's. PMID- 18411993 TI - New IBS guidance focuses on improving diagnosis and care. PMID- 18411994 TI - Patient assessment. Part 3--measurement of gastric fluid pH. PMID- 18411996 TI - Illicit drug use and its effect on the lungs. AB - Charlotte Rayner and Samantha Prigmore describe the prevalence of illicit drug use, how these drugs are inhaled or injected and the pulmonary consequences of such drug use. PMID- 18411995 TI - Chronic kidney disease anaemia. 2: Overview of guidance. AB - This is part 2 of a two-part unit on managing anaemia of chronic kidney disease (CKD). Part 1 outlined its prevalence, causes, signs and symptoms, and screening for it. Part 2 presents key points from published guidance and a summary of treatment options, and discusses nurses' role in managing people who have anaemia in CKD. PMID- 18411997 TI - How a nurse-led clinic cut outpatient waiting times. AB - Joe Annandale describes how changing the respiratory nurse specialist role can improve outpatient waiting times and meet patients' needs. PMID- 18411998 TI - Nursing patients with interstitial lung disease. AB - Annette Duck considers the important factors for supporting patients with a diagnostic label of interstitial lung disease. PMID- 18411999 TI - Striking the right balance. PMID- 18412000 TI - Leaving in good time. PMID- 18412001 TI - The ancient art of leadership. PMID- 18412002 TI - Knowing the score. PMID- 18412003 TI - Co-ordinating care. PMID- 18412004 TI - How clinical champions can improve quality. PMID- 18412005 TI - Essence of Care: a collaborative audit. PMID- 18412006 TI - Making good decisions: part 2. PMID- 18412007 TI - [The feto-fetal transfusion syndrome]. PMID- 18412008 TI - [Mental retardation and sexuality. Interview with book author Ilse Achilles (interview by Angelika Staub)]. PMID- 18412009 TI - [Munich coordination center of pediatric palliative medicine--a model for the future?]. PMID- 18412010 TI - [Brain death and organ donation in childhood--an explanation approach]. PMID- 18412011 TI - [Early intervention by pediatric nurses]. PMID- 18412012 TI - [Home parenteral nutrition of children with short bowel syndrome]. PMID- 18412013 TI - [Pesticide residues on fruit and vegetables--a problem for the consumer?]. PMID- 18412014 TI - [Unlikely neighbors. Hospice was built next to kindergarten]. PMID- 18412015 TI - [A virus as "global player". Chronic hepatitis B infection: frequently diagnosed too late--World Hepatitis Day October 1]. PMID- 18412016 TI - [Grades in nursing and pediatric nursing examination]. PMID- 18412017 TI - [Incidence of severe medical errors in intensive care medicine: results of the Critical Care Safety Study]. PMID- 18412018 TI - Reversal of GABA-mediated inhibition of the electrically and potassium chloride evoked [3H]-GABA release from rat substantia nigra slices by DL-3-hydroxy-3 phenyl pentanamide. AB - The phenyl alcohol amides, DL-2-hydroxy-2-phenyl butyramide (CAS 52839-87-9), DL 3-hydroxy-3-phenyl pentanamide (CAS 131802-69-2, DL-HEPP) and DL-4-hydroxy-4 phenyl hexanamide (CAS 67880-30-2) and their fluorine and chlorine analogs, at a concentration of 100 micromol/L, did not displace [3H]-gamma-aminobutyric acid ([3H]-GABA, CAS 108158-36-7) from GABAA receptors and only weakly displaced [3H] GABA and [3H]-CGP62349 (CAS 186986-97-0), a GABAB receptor antagonist, from GABAB receptors in rat brain crude synaptic membranes. The electrically and potassium chloride (15 mmol/L) evoked [3H]-GABA release in the presence of DL-HEPP, GABA and GABAB receptor ligands from rat brain substantia nigra (SN) slices was studied. R-Baclofen (CAS 69308-37-8) (10 micromol/L), a GABAB receptor agonist, produced an inhibition of the electrically evoked [3H]-GABA release and this inhibition was blocked by CGP 55845A (CAS 149184-22-5) (10 micromol/L), a GABAB receptor antagonist, but was not affected by DL-HEPP (100 micromol/L). CGP 55845A (10 micromol/L) did not alter the electrically evoked [3H]-GABA release in the absence of baclofen. The addition of DL-HEPP (100 micromol/L) alone did not affect the electrically-evoked release of [3H]-GABA release control, but it was able to significantly reduce the inhibitory effect of GABA (CAS 56-12-2) (10 micromol/L) on [3H]-GABA release evoked both by electrical and potassium chloride stimulation, in the presence of tiagabine (CAS 115103-54-3) (10 micromol/L), a GABA uptake blocker. In three preliminary experiments, bicuculline (CAS 485-49-4) (10 micromol/L) and picrotoxinin (CAS 17617-45-7) (10 micromol/L), two GABAA antagonists, inhibited the electrically evoked release of [3H]-GABA from rat SN slices, and DL-HEPP (100 micromol/L) reversed this inhibition. The mechanism of action of DL-HEPP is not known but, it might act as a negative GABA modulator in rat brain slices. PMID- 18412019 TI - Absorption, protein binding, pharmacokinetics and excretion of the anti-ischemic and anti-hypertensive arylpiperazine derivative CDRI-93/478 in rats. AB - CDRI-93/478 (1- [4-(4-fluorophenyl) piperazine-1-yl]-3-(2-oxopyrrolidin-1-yl) propane hydrochloride, an arylpiperazine derivative, is a potent anti-ischemic and anti-hypertensive agent and is in advanced stage of preclinical trials. In order to develop CDRI-93/478 into a clinical agent, the absorption, protein binding, pharmacokinetics, and excretion of the compound were investigated in male Sprague-Dawley rats. Oral absorption was evaluated in situ and in vivo, using the portal-venous concentration difference method. The compound showed negligible absorption (ka = 0.01 h(-1)) at pH 2.6. However, the rate of absorption of the compound at pH 7.4 was 0.6 h(-1) and was comparable to that observed in the in vivo study (ka, >0.58 h(-1)) in rats after a single 2 mg/kg oral dose. In vitro and in vivo protein binding studies using the ultrafiltration method showed that the compound was subject to low protein binding (<40%) and was independent of the substrate concentration over a range of 1-16 microg/ml. Pharmacokinetic parameters of the compound were determined after intravenous and oral administration of 0.6, 2 and 8 mg/kg doses using a model independent method. After oral administration, the compound showed the double-peak phenomenon, which could be due to the high water solubility (log P, 1.01 +/- 0.01), regional differences in the gastrointestinal absorption and enterohepatic recirculation effects. The absorption of CDRI-93/478 was rapid and showed a bioavailability of 69.9 +/- 5.1% (mean +/- S. D.) after 2 and 8 mg/kg oral dose. However, the pharmacokinetic parameters of the compound could not be determined after the 0.6 mg/kg oral dose due to insufficient data points. The studies following intravenous and oral administration demonstrated linear pharmacokinetics, low clearance and high volume of distribution over the dose range studied. The excretion studies after the 8 mg/kg oral dose indicated that the compound was not excreted through the feces and the urinary excretion was very low (<2%). PMID- 18412020 TI - Nitric oxide-independent activation of soluble guanylate cyclase by BAY 60-2770 in experimental liver fibrosis. AB - Liver cirrhosis is a chronic disease with high mortality rate and need for effective pharmacological intervention. The fibrotic remodelling of liver tissue is crucially dependent on hepatic stellate cell activation. Activation of hepatic stellate cells is reduced by an increase in cyclic guanosine monophosphate (cGMP). Stable cGMP analogues also reduce the contractile response of hepatic stellate cells. However, cGMP production is downregulated in the cirrhotic liver due to the reduced activity of the endothelial nitric oxide synthase. OBJECTIVE: Here we report that the novel activator of soluble guanylate cyclase (sGC), BAY 60-2770 (4-({(4-carboxybutyl) [2- (5-fluoro-2-{[4'-(trifluoromethyl) biphenyl-4 yl]methoxy}phenyl)ethyl] amino}methyl)benzoic acid), which increases the activity of sGC in a nitric oxide-independent manner, attenuates liver fibrosis in two rat models. METHODS: The compound was studied in the pig serum model and the carbon tetrachloride model. Fibrosis was assessed by estimating the increase in fibrous collagen by micromorphometry of histological sections stained with Sirius Red/Fast Green and by measuring total hepatic collagen. RESULTS: BAY 60-2770, on a recombinant sGC reporter cell line, stimulated the luminescence signals with an EC50 value of 5.4 +/- 1.2 nmol/L. In the presence of [1,2,4]oxadiazolo[4,3 a]quinoxalin-1-one (ODQ; 10 pmol/L) the EC50 was shifted to 0.39 +/- 0.11 nmol/L. In both fibrosis models, once daily oral administration of BAY 60-2770 concomittantly with the fibrotic stimulus prevented 60-75% of fibrosis, the lowest effective dose being 0.1 mg/kg in the pig serum model and 0.3 mg/kg in the carbon tetrachloride model. The treatment was well tolerated by all animals. The doses used were devoid of any significant influence on systemic blood pressure. CONCLUSION: Nitric oxide-independent activation of sGC might be an innovative therapeutic approach for the treatment of liver fibrosis of necro-inflammatory and immunological origin. PMID- 18412021 TI - Effects of D-004, a lipid extract from the royal palm (Roystonea regia) fruits, tamsulosin and their combined use on urodynamic changes induced with phenylephrine in rats. AB - 5-alpha-Reductase inhibitors, alpha1-adrenoreceptors blockers and herbal drugs, like lipid extracts from saw palmetto fruits, are used to treat benign prostatic hyperplasia (BPH). D-004, a lipid extract from the Royal palm fruits, prevented prostate hyperplasia (PH) induced with testosterone and the atypical PH induced with phenylephrine (PHE) in the rat, its effect in this last model being comparable to that of saw palmetto, but lesser than that of tamsulosin (CAS 106133-20-4). It was investigated whether single doses of D-004, tamsulosin and their combined therapy can prevent urodynamic changes induced with PHE in the rat. Firstly, the effects of PHE on rat volume voided per micturition (VM) were explored in rats that were distributed in three groups: a negative control and two groups injected s. c. with PHE (5 and 10 mg/kg, respectively). In the other two experiments, rats were distributed in four groups: a negative control and three groups injected with PHE (a positive control and two groups treated with either tamsulosin 0.05 and 0.1 mg/kg, or D-004 400 and 800 mg/kg. In another experiment, the effects of the combined therapy were assessed using four groups: a negative control, a positive control and three groups treated orally with tamsulosin 0.05 mg/kg, D-004 400 mg/kg or D-004 400 mg/kg + tamsulosin 0.05 mg/kg, respectively. Sixty min later, all rats (except negative controls) were injected s. c. with PHE (5 mg/kg), and all (including the negative controls) received a fluid-loading dose. Thirty min later, they were placed in metabolic cages and theVM was measured for 1 h. The VM was significantly reduced with PHE (5 and 10 mg/kg), the high dose producing anuria in 50% of the rats. The reduction of VM was significantly and dose-dependently prevented with tamsulosin (0.05 and 0.1 mg/kg) (42.9% and 60.3%, respectively) and with D-004 (400 and 800 mg/kg) (25.2% and 43.1%, respectively). The inhibition reached (70.9%) with the combined therapy was greater than that reached with each monotherapy and also greater than the sum (56.8%) of the inhibitions reached with tamsulosin (35%) or D-004 (21.8%) alone. In conclusion, tamsulosin (0.05 and 0.1 mg/kg) and D-004 (400-800 mg/kg) dose-dependently inhibited the VM reduction induced with PHE, their combined therapy producing the greater effects. PMID- 18412022 TI - Anti-tumor-Promoting activity of tibolone and its metabolites. AB - The aim of this study was to evaluate the cancer chemopreventive potential of the widely prescribed drug tibolone (17alpha-ethynyl-7alpha-methyl-5(10)-estren-3 one, CAS 5630-53-5) and its main metabolites, 17alpha-ethynyl-7alpha-methyl-4 estren-3-one (CAS 1162-60-3), 17alpha-ethynyl-7alpha-methyl-5(10)-estrene 3alpha,17beta-diol (CAS 100239-44-9) and 17alpha-ethynyl-7alpha-methyl-5(10) estrene-3beta,17beta-diol (CAS 100239-45-0), by studying their anti-tumor promoting activity. To this aim the test compounds were submitted to the short term in vitro assay for the inhibition of Epstein-Barr virus early antigen (EBV EA) activation induced by the tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA) as a primary screening for anti-tumor promoters. All the compounds showed high inhibitory activity and low cytotoxicity as compared to literature data. To extend the study to an animal model, tibolone and its 3alpha-hydroxy metabolite (CAS 100239-44-9) were also assayed in the in vivo two-stage on mouse skin carcinogenesis test, exhibiting significant inhibitory effects on TPA promoted mouse skin papillomas formation. A comparison with literature data indicated them as more potent compounds than other steroids previously studied such as digitoxigenin, cortisone, hydrocortisone, and prednisolone. PMID- 18412023 TI - Determination of ampicillin in human plasma by solid-phase extraction-liquid chromatography-tandem mass spectrometry (SPE-LC-MS/MS) and its use in bioequivalence studies. AB - A simple, fast, sensitive and selective solid-phase extraction-liquid chromatography-tandem mass spectrometry (SPE-LC-MS/MS) method for the quantitative analysis of ampicillin (CAS 69-53-4) in human plasma was developed using amoxicillin as internal standard, and sample extraction by solid-phase extraction (SPE). Extracts were separated by reversed-phase C18 with aqueous mobile phase (acetonitrile, 80:20, v/v) with 0.1% formic acid. The method was validated and successfully applied in a bioequivalence study of capsules 500 mg of ampicillin. Using a short running time of 2.5 min, the lower limit of quantification (LLOQ) for obtained ampicillin was 0.1 microg/ml for a plasma sample of 250 microl and a recovery of 94.38% +/- 4.05. Bioequivalence between the products was determined by calculating 90% confidence intervals (CI) for the ratio of Cmax, AUC0-t and AUC0-inf values for the test and reference products, which were within the 0.80-1.25 interval proposed by FDA and EMEA. It is concluded that the two formulations are bioequivalent in their rate and extent of absorption, and thus, may be used interchangeably. PMID- 18412024 TI - Comparative bioavailability study of two cefixime formulations administered orally in healthy male volunteers. AB - The bioavailability of a new cefixime ((6R,7R)-7-[(Z)-2-(2-amino-4-thiazolyl)-2 (carboxymethoxyimino) acetamido]-8-oxo-3-vinyl-5-thia-1-azabicyclo-[4,2,0]-oct-2 ene-2-carboxylic acid, CAS 79350-37-1) tablet preparation (Loprax) was compared with that of a reference preparation of the drug in 24 healthy male volunteers. The trial was designed as an open, randomized, single-blind, two-sequence, two period crossover study. Under fasting conditions, each subject received a single oral dose of 400 mg cefixime tablet as a test or reference formulation on 2 treatment days. The treatment periods were separated by a one-week washout period. The plasma concentrations of the drug were analyzed by a rapid and sensitive HPLC method with UV detection. The pharmacokinetic parameters included AUC0-24h, AUC0-infinity, Cmax, t1/2, and Ke. The mean AUC0-infinity of cefixime was 45008.7 +/- 10989.9 and 45221.3 +/- 2155.7 n x h/ml for the test and reference formulation, respectively. The maximum plasma concentration (Cmax) of cefixime was on average 4746.9 +/- 1284 ng/ml for the test and 4726.3 +/- 1206.9 ng/ml for the reference product. No statistical differences were observed for Cmax and the area under the plasma concentration-time curve for test and reference tablets. The calculated 90% confidence intervals based on the ANOVA analysis for the mean test/reference ratios of Cmax, AUC0-infinity and AUC0-24h of cefixime were in the bioequivalence range (94%-112%). Therefore, the two formulations were considered to be bioequivalent. PMID- 18412025 TI - Keel bone condition in laying hens: a histological evaluation of macroscopically assessed keel bones. AB - The objective of the present study was to conduct a corresponding histological analysis of 162 macroscopically assessed keel bones (1: severe, 2: moderate, 3: slight, 4: no deformity). Four layer lines were used and hens were kept in furnished cages, small group systems (both allowing more activities due to the provision of perches) and an aviary system, which fully conformed to the EU standards. Investigations were carried out in the 3rd, 6th, 9th and 12th laying month of two experimental trials. In 97.9% of grade 4 keel bones, no histological deviations were found, whereas in keel bones manifesting deformities of grade 1 and 2, the predominant histological observation was the incidence of fracture callus material (FCM) and new bone in the form of woven bone. FCM was also detected in 50.9% of grade 3 keel bones, whereas in 40.7%, only s-shaped deviations of keel bones were found, which were related to extended pressure loading while perching activities rather than short-duration trauma. Histological analysis showed that keel bones of grade 1 and 2 were mainly attributed to traumatic origin and therefore associated with pain experience in layers. Grade 3 keel bones manifested either FCM as a result of trauma or adaptational deformities without any evidence of a preceding fracture in response to mechanical pressure loading and were most likely not associated with pain. Therefore, histological analysis was found to be a mandatory tool when evaluating grade 3 keel bones with respect to layers'welfare. Furthermore, this analysis corroborates the findings that in aviary systems deformities of keel bones are predominantly caused by painful fractures. PMID- 18412026 TI - Is moisture really the most important influencing factor on the mechanical properties of claw horn? Modulus of elasticity and dry-matter content in flat and contracted claws. AB - The modulus of elasticity (E) and dry matter content (DMC) of horn samples from two types of claw deformity (contracted and flat claws) were determined according to EN-ISO 527 and ASTM D 638-03. The overall E values for all segments combined were 168 +/- 154 MPa (mean +/- SD, n = 79) and 248 +/- 196 (n = 90) for contracted and flat claws, respectively. These values were approximately 60% and 40% less than the E value of sound claw material. The high correlation between E and DMC, as previously found for sound claw horn material was not present in horn obtained from these types of pathologically altered claws. PMID- 18412027 TI - Genetic and phenotypic trends in canine hip dysplasia in the German population of German shepherd dogs. AB - Phenotypic and genetic trends were examined in 157,455 German shepherd dogs of the German population born between 1985 and 2003 which were Xrayed for canine hip dysplasia (CHD). A subset of 40,270 dogs born between 1998 and 2003 was examined for which information was available on age at examination and X-raying veterinary practitioner. Relative breeding values were predicted using best linear unbiased prediction methods (BLUP) and Gibbs Sampling. There was an almost linear and significant decline in both phenotypic and genetic trends for CHD scores (scale 1 to 5 for CHD free to severe CHD) over a period of 19 years, with b = -0.021 +/- 0.002, and b = -1.69 +/- 0.05, respectively. After introduction in 1999 of a selection scheme based on relative breeding values (RBVs, mean 100, standard deviation 20) of a linear animal model, phenotypic and genetic trends declined less than before 1999, with b = -0.019 +/- 0.011 and b = -1.43 +/- 0.29. Analysis of the effects of RBVs of parental combinations among sires and dams on the percentage of non-affected progeny showed that lower RBVs were associated with a higher percentage of non-affected progeny than were higher RBVs. In addition, it was also shown that selection procedures based on RBVs are more efficient than selection schemes based on phenotypic records of parents. When the maternal RBV was above 100, the proportion of non-affected progeny was between 10% and 20% lower than when the maternal RBV was below or equal to 100. There was a significant relationship between higher inbreeding coefficients and higher RBVs of the animal. PMID- 18412028 TI - [Use of GnRH in livestock production--current position and new research data]. AB - The hypothalamic gonadotropin-releasing hormone (GnRH) is seen as the key hormone of neuroendocrine regulation of reproduction. The ability of GnRH and its analogues to stimulate the release of the gonadotropins FSH and LH is world-wide utilized for various veterinary purposes, including treatment of certain hormone dependent disturbances and stimulation of ovulation in controlled breeding programmes. A large difference is striking, however, when comparing the efficiencies reported. This may underline the importance of accurate treatment and reflect the manifold influences by animals and their environment on reproductive performance. During the last years, novel analytical methods have been established enabling a significant progress in reproductive research. The discovery and characterization of natural GnRH variants and their receptors in several vertebrate species may become more important. The reason is, that these GnRHs affect the release of the gonadotropins FSH and LH, but they may transmit, moreover, seasonal and nutritive signals to reproductive organs. It might be expected that the further exploration of these functions may serve as basis for the development of new and effective biotechnical methods in farm animal treatment. PMID- 18412029 TI - The development of microflora and production of short-chain fatty acids in the digestive tract of suckled piglets and replacer-fed piglets. AB - The objective of this study was to obtain knowledge about the postnatal development of microflora and the production of short-chain fatty acids in 24 piglets suckled by sows and 26 piglets fed on milk replacement. On day 14 piglets which had received no colostrum had higher counts of Enterobacteriaceae (p < 0.001) and coliform bacteria (p <0.001) in the jejunum contents than piglets suckled by their mother sow. Depending on age, concentrations of both lactic and acetic acid were higher in the contents of the small intestine of piglets suckled by sows compared to milk replacer-fed piglets. Replacement of maternal milk by artificial feeding adversely affected the postnatal development of the piglets. This resulted in higher morbidity and mortality in those piglets. PMID- 18412030 TI - Effects of different crating periods on selected biochemical indices in broiler chickens. AB - Effects of different crating periods on selected biochemical indices were monitored in a group of unsexed ROSS 308 broiler chickens aged 42 days. 120 randomly selected chickens were weighted and placed in 15 heavy duty plastic crates with reduction in floor space approx. 160 cm2/kg of body weight of broilers (average body weight was 2.75 kg). Five crates were kept for 4 hours (Group 4h), another five crates for 8 hours (Group 8 h) and another five crates for 12 hours (Group 12h). After tested periods of crating the broilers were released and blood samples were withdrawn from Vena bosilico in 24 randomly selected birds from each group (Groups 4 h, 8 h, 12 h respectively) and also in other 24 randomly selected non-crated control birds. The results of our experiment proved that duration of crating significantly influenced the stress response in broiler chickens. The duration of crating period was positively correlated with the plasma corticosterone level and negatively correlated with the lactate dehydrogenase, cholesterol, triglycerides, glucose and lactate levels in the blood of broiler chickens. PMID- 18412031 TI - [Influence of subclinical pulmonary findings on cardiac parameters in Icelandic horses]. AB - In the present study we examined, if in Icelandic horses an increase in heart and/ or breathing rate is physiological and breed dependend or a sign of a pulmonary or cardiac disease. Therefore we examined 37 Icelandic horses with the prereport of being healthy. During clinical lung examination four horses showed symptoms of a pulmonary disease like increased breathing rate and enforced breathing at rest. These horses were excluded from the study. The other 33 horses were clinically normal. 17 of these horses were unridden (untrained) and 16 horses were regularly worked (trained). After clinical examination in all horses analysis of arterial blood gas, endoscopy with tracheo- bronchial secret analysis and radiographic examination of the lung were carried out. Additionally electro- and echocardiographic examinations and standardised exercise tests with determination of heart and breathing rate as well as plasma lactate values were performed in all horses. During electro- and echocardiographic examination no pathological findings were observed. In total 22 of the 33 horses showed abnormal lung findings. Seven horses had mild signs of RAO and 15 horses had mild signs of interstitial bronchitis. Three horses had additional pulmonary haemorrhage. Eleven out of the 33 horses showed no abnormal lung findings. The breathing rate at rest differed not significantly between horses with (21 +/- 1/min) or without (23 +/- 2/min) pulmonary findings. The heart rate also did not differ significantly between horses with (39 +/- 1/min) or without (42 +/- 1/min) pulmonary findings. In contrast to this the trained Icelandic horses with abnormal pulmonary findings had significantly higher heart rates (p = 0.01) and significantly lower breathing rates (p = 0.009) compared to those without abnormal pulmonary findings. During echocardiography Icelandic horses with abnormal pulmonary findings had significantly larger left atrial diameter (without abnormal pulmonary findings: 82 +/- 7 mm, with abnormal pulmonary findings: 90 +/- 8 mm, p = 0.02). Compared to the untrained Icelandic horses (5.4 +/- 2 mmol/l) the trained horses showed significantly lower plasma lactate values (3.1 +/- 2 mmol/l, p = 0.001) immediately after exercise. After exercise the icelandic horses with abnormal pulmonary findings had significantly higher breathing rates (p < 0.05) and longer recovery periods (30 minutes) than horses without abnormal respiratory findings (15 minutes). Recovery of heart rate after exercise showed no differences between groups. PMID- 18412032 TI - [Ultrasonographic findings of disorders of the tarsal region in 97 cattle- arthritis, bursitis, tenosynovitis, periarticular abscess and vein thrombosis]. AB - The clinical, radiographic, ultrasonographic and centesis findings of 97 cattle with disorders of the tarsal region (arthritis, bursitis, tenosynovitis, abscess, vein thrombosis) were reviewed (1994-2006), and the typical ultrasonographic findings of different tarsal disorders in cattle are presented. The following criteria were assessed: echogenicity, echopattern, size, shape, appearance of the borders of synovial or other cavities. Ultrasonographic findings were compared with the clinical, radiographic and arthrocentesis results and with intraoperative and/or necropsy findings. 50 cattle suffered from one tarsal disease only, in 47 cattle--up to three different--concurrent disorders in one tarsal region were identified. The afflicted synovial cavities and abscesses were definitely differentiated ultrasonographically in all cattle suffering from tarsal disorders a comprehensive diagnosis could be achieved due to the easy and rapid ultrasonographic differentiation of the incriminated soft tissues. PMID- 18412033 TI - [Canine intervertebral disk disease: a retrospective study of clinical outcome in 238 dogs (2003-2004)]. AB - The aim of the study was the assessment of the prognosis of intervertebral disk diseases on the basis of a large case load with the help of currently available therapy modalities. Medical records of 238 dogs presented within 2 years (2003 2004) with neurological deficits resulting from intervertebral disk disease and surgical treatment at the Department of Small Animal Medicine and Surgery of the University of Veterinary Medicine Hannover were reviewed. 85% of the patients showed a good and 15% a poor long-term functional outcome. Both the presence of deep pain sensation and the weight of the dog were the only variables detected as having a significant impact on prognosis. The time needed to motor function improve depended on the degree of neurological deficits before surgery, and patients with a cervical lesion had a more rapid recovery than patients with a thoracolumbal disease. All other variables, including age, previous signs of intervertebral disk disease, preoperative steroid medication, a lower motor neuron involvement and durotomy did not significantly influence the outcome. Radiographic changes of the vertebral column such as spondylosis, spondylarthrosis, a narrowed disk space up to a fusion of the vertebral bodies mostly were found at the site of a removed intervertebral disk. In dogs with intervertebral disk disease evaluated in the current study a good result could be obtained after surgical decompression. PMID- 18412034 TI - A survey of subjective sleepiness and consequences in attending physicians. AB - This article surveyed attending physicians on their work hours, sleep schedule, daytime sleepiness, and the perceived relation of these factors to patient safety, quality of care, and personal well-being. Physicians answered demographic and workload questions and attitudinal questions regarding work-hour limitations; the Epworth Sleepiness Scale (ESS) was used to measure subjective sleepiness, and an 18-item Impact Questionnaire was also used. Of 180 participants, 41 (23%) attending physicians manifested abnormal ESS scores (11 or greater). Private practice- and surgically-based subspecialties had higher ESS scores. Reduced sleep, but not hours worked, was associated with increased sleepiness. Sleepy physicians were more likely to associate sleep loss with medical errors and driving impairment. Sleepiness may be attenuated by education regarding consequences of insufficient sleep and institution of effective countermeasures. PMID- 18412035 TI - University students and "the all nighter": correlates and patterns of students' engagement in a single night of total sleep deprivation. AB - This study evaluated the self-reported patterns, motivations, and correlates of engagement in a single night of total sleep deprivation (SN-TSD) in a sample of 120 university students at a 4-year liberal arts college. Participants' mean age . was 20.1 years; the sample was 63% female. Measures included the Owl-Lark Scale of circadian preference; the Beck Depression Inventory; and the Pittsburgh Sleep Quality Index (PSQI); as well as self-reported usual bedtime, rise time, and total sleep time. Students' cumulative grade point averages (GPAs) were also obtained; these were reported from the university registrar. Sixty percent of students surveyed reported engaging in SN-TSD once or more since coming to college. No differences between genders or among class standing with respect to use of SN-TSD were detected. Engagement in SN-TSD was associated with later self reported bedtimes, evening preference, and poorer academic achievement as measured by GPA. A trend for increased symptoms of depression was detected; sleep quality as measured by the PSQI was not related to engagement in SN-TSD. PMID- 18412036 TI - Chronic insomnia and cognitive functioning among older adults. AB - Chronic insomnia and cognitive impairment are both common complaints among older adults. This study explores the association between chronic insomnia and changes in cognitive functioning among older adults. The study population comprised two groups: 64 older adults without insomnia and 35 older adult insomniacs. The cognitive capacity of each participant was tested at the participant's home using the computerized "MindFit" test (CogniFit, Inc.). In five categories of cognitive functioning, older adult insomniacs displayed impaired performance compared to older adult good sleepers. Specifically, significant differences were found between insomniacs and good sleepers on memory span, allocating attention to a target, time estimation, executive functioning, and integration of two dimensions (visual and semantic) tests. The findings imply that insomnia may have detrimental effects on some cognitive functions in healthy older adults. PMID- 18412037 TI - Circadian phase in sleep-disturbed adolescents with a history of substance abuse: a pilot study. AB - The relationship between actigraphy- or diary-based sleep parameters and salivary melatonin-based dim light melatonin onsets (DLMOs) was examined in 21 adolescents with a history of substance abuse and current complaints of sleep difficulties. The adolescents displayed relationships between diary-based sleep times and DLMO that were of comparable strength with those reported for adult insomniacs and healthy adolescents during the school year, but weaker than those observed in healthy adults and healthy adolescents on summer vacation. When the sample was divided into adolescents with late and early DLMOs, the 2 groups had significantly different phase angles between DLMO and sleep variables but no other significant differences in sleep parameters. However, circadian phase and its relationship to sleep may have sleep and behavioral consequences. PMID- 18412038 TI - Toxoplasma gondii infection in horses. A review. AB - This review updates those written by Dubey and Beattie in 1988 (1988a) and by Tenter et al in 2000, on pathological and epidemiological aspects of Toxoplasma infection in horses. Under natural conditions, seroprevalence may variate from 0% up to 90%. This wide variation may be due to the sensitivity of the serological methods, to the age of animals, to the geographical area, and even to the hygienic condition of the farms and farm management. With few exceptions, horses are considered one of the less sensitive specie to the pathogenic effect of Toxoplasma gondii. In fact, neither under experimental nor under natural condition a genuine pathologic picture related to the toxoplasmic infection has been described. In one occasion the organism has been isolated from an eye condition and in others a connection between a higher frequency of unspecified pathological conditions and a positive response to serological test for Toxoplasma has been speculated. Diaplacental transmission and the following abortion have been only occasionally reported, and at least in one case in a quite trustworthy way, therefore it must be considered possible, though rare. Although infection of humans due to the consumption of horse meat has never been reported, the existence of a possible risk arouses by the demonstration of the presence of parasite stages in either naturally or experimentally infected horses, which resulted to be infective for mice and/or cats. PMID- 18412039 TI - Importance of phoresy in the transmission of Acarina. AB - Dispersal capacity plays a central role in the radiation of animals, facilitating the exploitation of habitats variously distributed in space or in time or both. Many living species are unable to leave a host, crawl around, and find a new host, so they must rely on external factors to be transmitted. Biotical factors may be important in passive transport and the process, by means of which an animal is passively transported by a selected carrier of different species, is known as "phoresy". Phoresy is a phenomenon in which one animal (the phoretic) seeks out and attaches to an animal of another species, with which it does not share any phase of the life cycle, for dispersal, during which time the phoretic animal becomes quiescent, stopping feeding and development. Activity starts again beginning with detachment, induced by stimuli originating from its carrier or the microhabitat. The adaptive traits of phoresy may be categorized as follow: host surface, quiescence, recognition of signals to abandon the carrier and, if needed, synchronization with the host life cycle. Phoresy is exploited by many Arthropods. In Acarina, there are basically four main types of phoresy. First, there is a type in which adult females are the only forms becoming phoretic and attachment is by means of chelicerae, palpal hooks and ambulacral claws, which grasp a seta or a fold of the integument of carrier-host. The second type is represented by mites, in which deutonymphs are phoretic; there is generally no cheliceral or sucker attachment in this group, mites instead hanging on by their ambulacral claws. The third type is similar to the second in that deutonymphs are phoretic; however, in this case, attachment to the host is by means of an anal pedicel formed by a substance, extruded through the anus, which hardens upon coming in contact with air and literally glues the mite to its host. In the fourth type there is a very highly modified deutonymph stage, called hypope, which only occurs at certain times, presumably when environmental conditions are no longer appropriate for the mite. Hypope is simplified morphologically, may have many sucker-like discs or claspers for efficient attachment, and is much more resistant to desiccation than are other stages of the life cycle. PMID- 18412040 TI - A comparison of five methods for DNA isolation from liver and rumen flukes to perform ITS-2+ amplification. AB - Five different DNA isolation methods (4 commercial kits and a modification of phenol-chloroform method) were compared for the discrimination of adults of Fasciola hepatica and Dicrocoelium dendriticum (liver flukes), and Calicophoron daubneyi (rumen fluke) collected from sheep in southern Italy. The second internal transcribed spacer (ITS-2) of ribosomal DNA (rDNA) plus flanking 5.8S and 28S sequence (ITS-2+) was amplified by polymerase chain reaction (PCR) from serial diluted DNA templates (6 ng - 60 fg) of each fluke species. Overall, in terms of efficiency in detection limit, the best results were obtained either with phenol-chloroform purification or with QIAamp DNA Mini Kit (Qiagen), but using this latter method, rapid, safe and not expensive, an increased level of sensitivity sufficient to detect small amounts of target-DNA was achieved. In addition, electrophoresis analysis following PCR also showed that ITS-2+ could be useful as a genetic marker for the molecular identification of F. hepatica, D. dendriticum and C. daubneyi in definitive and intermediate hosts. Furthermore, for the first time, the ITS-2 sequence of D. dendriticum was defined. PMID- 18412041 TI - Case report: effects of diethylcarbamazine and thiabendazole combination against Mansonella perstans filariasis. AB - Mansonella perstans filariasis is widely present in Africa and equatorial America and its pathogenicity has been recently reconsidered. Effective treatment is lacking and there is no consensus on optimal therapeutic approach. We present the results of a new combination treatment against M. perstans filariasis. Two cases of M. perstans filariasis were treated with the combination of diethylcarbamazine (DEC) and thiabendazole. The treatment was able to significantly reduce microfilaria burden in a case and to achieve complete clearance of blood microfilariae in another case. PMID- 18412042 TI - A simple duplex-PCR protocol for routine diagnosis and follow up of canine leishmaniasis. AB - The introduction of PCR has efficiently improved diagnosis of canine leishmaniasis. In order to provide a robust, efficient and reliable diagnostic method, a duplex PCR assay targeting the Leishmania infantum kDNA minicircle and the canine GAPDH gene as inner control was designed. Sensitivity of the assay reached 0.15 parasites/ml blood. Development, testing and application of this system on a group of 10 dogs during therapy administration (60 days) are also described. Six dogs (out of eight that have been showing a positive PCR result on peripheral blood during the study) were tested negative at day 62, indicating a reduction of parasitaemia at the end of the treatment period. All the animals had a positive PCR on lymph node aspirate both at the beginning and at the end of treatment. These findings seem to suggest that, in order to test therapy efficacy, PCR on whole blood could be a useful assay in dogs that have a positive PCR at the beginning of the treatment, while PCR positivity on lymph nodes lasts longer than the observation period during therapy administration. The presence of the GAPDH inner control band efficiently contributed to prevent false negatives, by highlighting samples affected by haemoglobin inhibition or inappropriate DNA isolation. PMID- 18412043 TI - Amebic liver abscess: report of three cases. AB - Amebic abscess is a common manifestation of extraintestinal amebiasis and it is associated with relatively high morbidity and mortality. We present three cases seen in Bari, Southern Italy, one of which was autochthonous and the other two were not. Diagnosis was performed by elevated antibody titre for E. histolytica through immunofluorescence assay and positive antigen determination by ELISA in stools and in abscess aspirate. Fever often accompanied by chills, abdominal pain, weight loss and hepatomegaly were present. Laboratory findings also revealed leukocytosis with neutrophilia. Pleural effusion was observed in two patients. In all our patients multiple abscesses were observed. All the patients were treated with metronidazole and two of them also underwent the aspiration of the amoebic abscess. In all of them there was improvement of the clinical picture, as demonstrated by computerized tomography. PMID- 18412044 TI - Blood parasites in birds from Burkina Faso. AB - This study aims at evaluating the prevalence and density of haemoparasites in a native population of immature and adult male and female birds of Burkina Faso. Protists from the orders Eucoccidiorida, Kinetoplastida, as well as microfilaria were found. During the period from the 27th to the 29th of November 2003, we trapped 110 birds: on 101 sampled birds, 47 harboured blood parasites, with an overall prevalence of infection of 46.5%; 15 birds (14.8%) harboured mixed infections. Haemoproteus (prevalence of infection 30.7%), Plasmodium (13.9%), Trypanosoma (10.9%), microfilaria (8.9%) and Leucocytozoon (5.9%) were most frequently recorded. Occurrence of blood parasites was very high in comparison to that seen in birds from other areas of Africa. PMID- 18412045 TI - Structural changes in the salivary glands of Ixodes ricinus larvae. AB - The authors analysed the structure of Ixodes ricinus (L.) larvae in specimens immediately after leaving the egg sheaths, in those which have not fed for 2 months after hatching, and in feeding larvae on the second day of feeding. The results showed that salivary glands in tick larvae are formed by alveoli aligned in strands on both sides of the central nervous system. These alveoli open into central efferent ducts via short ducts. The constituent elements of salivary glands include pyramidal alveoli (with numerous lipid droplets) and granular alveoli of varied structure. It is worth noting that salivary alveoli containing secretory material are present even in the larvae which had just left egg sheaths and were still endowed with deutoplasm. PMID- 18412046 TI - The genetic resistance to gastro-intestinal strongylids in Appenninica sheep: preliminary results. AB - Heritability parameters of resistance to gastro-intestinal strongylids, measured as FEC (Faecal Egg Count), were evaluated in the Appenninica sheep breed. FEC heritability coefficient was 0.11 +/- 0.061 while FEC repeatability coefficients were 0.58 +/- 0.085 and 0.76 +/- 0.223 in adult females and lambs respectively. Subjects were classified, based on FEC, into three different levels of resistance to strongylids. Ewes belonging to the 'resistant class' should be conveniently exploited in mating schemes, in order to provide a method, alternative to drug administration, for a long-term parasite control; this would result particularly helpful under those production systems, such as organic farming, where the use of drugs is not allowed or limited. PMID- 18412047 TI - Reappearance of Aedes albopictus in Sardinia. AB - Aedes (Stegomyia) albopictus is a vector of major human diseases. From 1979 established populations were found in the Mediterranean Region. The first record of Ae. albopictus in Italy goes back to 1990 and at present it is permanently established in several provinces of central and northern Italy. Ae. albopictus was detected for the first time in Sardinia in 1995, and a new record was reported in 1997. After these two isolated episodes, the "tiger-mosquito" was considered apparently eradicated. At the end of September 2006, two natural breeding populations including adults of Ae. albopictus were recorded in two different localities of South-Sardinia: in the botanical garden of downtown Cagliari and in a private garden of a small village, Teulada, on the south western coast of Sardinia. The origin of this new introduction of Ae. albopictus in Sardinia is still unclear. In both cases the species has been found in artificial containers in densely populated areas. PMID- 18412048 TI - Actinosporean stages of Myxozoan parasites of oligochaetes from Nera River basin, Central Italy. AB - The aim of this study was to check the occurrence of actinosporean stages of Myxozoan parasites of oligochaetes in the Nera River basin (Central Italy). The investigation was conducted near twelve trout farms (rainbow and brown trout) along the Nera River and in the littoral of the Piediluco Lake. Four actinosporean types, belonging to the collective groups aurantiactinomyxon, triactinomyxon, raabeia and echinactinomyxon, were detected in some of the biotopes investigated (Piediluco Lake and Sordo River). Differential diagnosis, carried out on a sub-sample of spores, showed a complete fitting with forms previously described in literature, with the exception of the triactinomyxon type. PMID- 18412049 TI - A checklist of Digenea parasitic helminths from the North Adriatic Sea. AB - The author presents an overview of isolated trematodes along their morphological description, isolated from the hosts that are for the first time reported in the North Adriatic Sea. The endohelminth parasites invasion of Northern Adriatic fish was studied during a 10-year research program. A total of 63 marine fish species of pelagic and bentic fish of the classes Chondrichthyes and Osteichthyes were analysed. Twenty-two fish species were found to host Digenean trematodes (33.33% prevalence), and 12 new host fish were reported for this class. During this endohelminths parasitofauna research work 63 species of sea-fish from Northern Adriatic (total of 2659 fish) were examined and 25.16% found invaded by some of endohelminths. The presence of 22 different species of flukes (Trematoda, Digenea) was found. Flukes were present in 33.33% of invaded fish. Furthermore 12 new fish hosts for Trematodes were found and this is the object of the present paper. The names of the investigated species of Trematodes together with those of their new hosts are listed in Table 1. The data referring to each parasite are given in the specific part of this study. PMID- 18412050 TI - New species of Choleoeimeria (Apicomplexa: Eimeriidae), coccidia of bile-bladders of reptiles, illustrating a multiplicity of host cell-parasite interrelations. AB - Oocyst characteristics and histological features of the endogenous development of bile-bladder coccidia of the genus Choleoeimeria Paperna and Landsberg, 1989 are described and the main features for species differentiation are discussed for the following new species: C. allogamae n. sp. from Agama sp., Cameroon, West Africa; C. allogehyrae n. sp. from Gehyra australis, Magnetic Island (type) and mainland N Queensland, Australia; C. boulii n. sp. from Gehyra variegata, SW Queensland, Australia; C. calotesi n. sp. from Calotes mystaceus, Xiang-Mai, Thailand; C. heteronotis n. sp. from Heteronotia binoei, N Queensland, Australia; C. lygosomis n. sp. from Lygosoma buringi, Kon-Kaen, Thailand; C. sylvatica n. sp. from Carlia rhomboidalis, N Queensland, Australia, and C. xiangmaii n. sp. from Hemidactylus frenatus, Xiang-Mai, Thailand. Oocyst characteristic of Choleoeimeria are also reported from Oedura castelnaui, N Queensland. The described species demonstrate a diversity of associations with the bile-bladder epithelial lining, from a single parasite in a single hypertrophic host cell to multiple infections inducing the hypertrophied cells to form stratified layers, or merge into branched clumps. PMID- 18412051 TI - A new pronocephalid, Pleurogonius tortugueroi n. sp. (Digenea), from the intestine of green sea turtles (Chelonia mydas) in Costa Rica. AB - A new species of trematode, Pleurogonius tortugueroi n. sp. (Digenea: Pronocephalidae) is described from the lower intestine of green sea turtles (Chelonia mydas) from Tortuguero National Park, Costa Rica. The new species differs from all other species of Pleurogonius by having a short oesophagus and oval testes close to lateral posterior limit of the body. It differs from all other species, except P. malaclemys Hunter 1961, by having an ovary between the testes; moreover the latter species is a parasite of freshwater turtles. All others members of the genus have a long oesophagus, testes placed to some distance from the posterior end, and the ovary is pretesticular. The new species appears most closely related to P. linearis Looss, 1901 but differs from it by having a different body shape, lappets of the head collar close at the cecal bifurcation level, a longer vitellarian field, different testis shape and position, ovary intertesticular, and different egg size. PMID- 18412052 TI - Parasites of the digestive tract in beef cattle imported from France to Italy. AB - Beef cattle heads (195 heads, 6 batches) imported for fattening from France to Italy were examined. Coprological qualitative and quantitative tests were performed, and the results analysed in relation to sex, breed, age, date of arrival, geographical origin (French department in which the animal was bred), and gathering centre (French department in which the animal was grouped with the others before travelling to Italy). The following parasites were identified: Eimeriidae (overall prevalence 60.5%); Strongyles (59%); Nematodirus spp. (14.3%); Trichuris spp. (4.1%); Capillaria spp. (2.0%); Paramphistomum spp. (27.6%); Dicrocoelium spp. (3.0%); Moniezia spp. (8.7%). All the observed parasites are widely reported in beef cattle either in Italy or in France. However, the seeming absence of Fasciola hepatica was unexpected, as well as the high prevalence of Paramphistomum spp. The variables that appeared to be more linked to parasite epidemiology were sex, altitude of the geographical origin and season. PMID- 18412053 TI - The cause of death of Emperor Charles V. AB - Abstract. Plasmodium falciparum malaria was the cause of death of Emperor Charles V. This confirmation was achieved by microscopy analysis of the Emperor's remnants. PMID- 18412054 TI - Cardiac CTA, three-dimensions, and the chronic total occlusion: a window to the future. PMID- 18412055 TI - Dosing radiation: limiting the invisible toxicity. PMID- 18412057 TI - The nosologic relationship between generalized anxiety disorder and major depression. AB - Generalized anxiety disorder (GAD) has undergone a series of revisions in its diagnostic criteria that has moved it, nosologically, away from its original affiliation with panic disorder (PD) and closer to major depressive disorder (MDD). This, together with its high comorbidity and putative shared genetic risk with MDD, has brought into question its place in future psychiatric nosology, prompting the planners of Diagnostic and Statistical Manual-V (DSM-V) and International Classification of Diseases-11 (ICD-11) to set up a workgroup tasked to better understand the relationship between GAD and MDD. This review attempts to summarize the extant data to compare GAD and MDD on a series of research validators to explore this relationship. Although insufficient data currently exist for GAD in several key validator classes, tentative conclusions can be drawn on the diagnostic status of GAD in relation to MDD. Although GAD possesses substantial overlap with MDD in the areas of genetics, childhood environment, demographics, and personality traits, this tends to hold true for other anxiety disorders (ADs) as well, with the strongest evidence for PD. Data from life events, personality disorders, biology, comorbidity, and pharmacology are mixed, showing some areas of similarity between GAD and MDD but some clear differences, again with a moderate degree of nonspecificity. Thus, although the bulk of evidence supports a close underlying relationship between them, the relatively nonspecific nature of these findings provides little more reason to question the nosologic validity of GAD in relation to MDD than that of some other anxiety disorders. PMID- 18412056 TI - Is generalized anxiety disorder an anxiety or mood disorder? Considering multiple factors as we ponder the fate of GAD. AB - Generalized anxiety disorder (GAD) and major depressive disorder (MDD) demonstrate a strong relationship to each other at both genotypic and phenotypic levels, and both demonstrate substantial loadings on a higher-order negative affectivity factor [see Watson, 2005: J Abnorm Psychol 114:522-536]. On the basis of these findings, there have been a number of calls to reclassify GAD in the same category as MDD (the "distress disorders"). However, any consideration of the reclassification of GAD should also take into account a number of other factors not only related to GAD and MDD but also to the overlap of these disorders with other anxiety and mood disorders. First, GAD has established reliability and validity in its own right, and specific features (e.g., worry) may become obscured by attempts at reclassification. Second, examination of the nature of the overlap of GAD and MDD with each other and with other disorders suggests a more complex pattern of differences between these conditions than has been suggested (e.g., MDD has strong relationships with other anxiety disorders, and GAD may be more strongly related to fear than it may first appear). Third, although findings suggest that GAD and MDD may have overlapping heritable characteristics, other evidence suggests that the two disorders may be distinguished by both environmental factors and temporal presentations. Finally, although overlap between GAD and MDD is reflected in their relationships to negative affectivity, temporal relationships between these disorders may be demonstrated by functional changes in emotional responsivity. PMID- 18412058 TI - Should OCD leave the anxiety disorders in DSM-V? The case for obsessive compulsive-related disorders. AB - Recently in 2006, a group of experts in obsessive compulsive disorder (OCD) and obsessive compulsive-related disorders (OCRDs) convened in Washington, DC, to review existing data on the relationships between these various disorders, and to suggest approaches to address the gaps in our knowledge, in preparation for the upcoming Diagnostic and Statistical Manual (Fifth Edition) (DSM-V). As a result of this meeting, the Research Planning Agenda for DSM-V: OCRD Work Group suggested removing OCD from the anxiety disorders, where it is currently found. This proposal is in accordance with the current International Classification of Mental Disorders (ICD-10) classification of OCD as a separate category from the anxiety disorders. Although the ICD-10 places both OCD and the anxiety disorders under the umbrella category of "neurotic, stress-related, and somatoform disorders," they are two separate categories, distinct from one another. As OCD and other putative OCRDs share aspects of phenomenology, comorbidity, neurotransmitter/peptide systems, neurocircuitry, familial and genetic factors, and treatment response, it was proposed to create a new category in DSM-V entitled OCRDs. Alternatively, the OCRDs might be conceptualized as a new category within the broader category of anxiety disorders. Future studies are needed to better define the relationships among these disorders, and to study boundary issues for this proposed category. There are both advantages and disadvantages in creating a new diagnostic category in DSM-V, and these are discussed in this article. PMID- 18412059 TI - Is disorder X in category or spectrum Y? General considerations and application to the relationship between obsessive-compulsive disorder and anxiety disorders. AB - Is obsessive-compulsive disorder (OCD) best categorized as an anxiety disorder? This question has been raised previously, but advances in the psychobiology of OCD and the anxiety disorders, and preparations for Diagnostic and Statistical Manual of Mental Disorders-V and International Classification of Diseases-11, make reconsideration timely. The debate in turn raises the more general issue of how best to address any question of the form "is disorder x in category or spectrum y?" Such questions are related to a number of key debates in philosophy of science and language and have also increasingly been addressed by the cognitive-affective neuroscience of categorization. Here, we review this background debate and use OCD as a relevant exemplar. PMID- 18412060 TI - Where does obsessive-compulsive disorder belong in DSM-V? AB - A reclassification of obsessive-compulsive disorder (OCD) into a new diagnostic category spectrum of "obsessive-compulsive spectrum disorders" (OCSDs) has recently been proposed, with considerable debate, for the forthcoming Diagnostic and Statistical Manual-Fifth Edition (DSM-V). This paper provides a critical analysis of the available empirical data regarding this conceptual and nosological shift. Specifically, we review research on shared commonalities and differences between OCD and the putative OCSDs in relation to their clinical presentation, phenotype, neurobiology, and treatment response. We conclude that a reclassification of OCD into a separate OCSD spectrum is premature and not supported by the currently available data. PMID- 18412062 TI - Behavioral inhibition. AB - Over the past 25 years, our understanding of the risks conferred by "behavioral inhibition to the unfamiliar" (BI) has grown tremendously, yet many questions remain. BI represents the persistent tendency to show extreme reticence, fearfulness, or avoidance in novel situations or with unfamiliar people. Prospective studies of high-risk offspring, selected community children, and unselected epidemiologic samples converge to suggest that BI confers specific risk for social anxiety disorder in early and middle childhood and adolescence. Later outcomes are less clear, with some studies suggesting associations with depression or panic disorder. Studies that find broad associations between BI and anxiety proneness in general may be limited by the absence of information about parental psychopathology (an important potential confound associated with both BI and anxiety disorders in offspring). A critical area for further inquiry is the degree to which BI confers risk for social anxiety disorder in the absence of family history of anxiety disorders. Additionally, although progress has been made in identifying risk factors, protective factors, and treatments that may influence the course of BI and associated anxiety, more work is needed. Also, several exciting inroads have been made into the genetic and neurobiologic underpinnings of BI, and future studies promise greater elucidation of these areas. For now, the clinical take-home message is that preschool-age children presenting with extreme and persistent BI are at elevated risk for social anxiety disorder and possibly for other future disorders; preliminary evidence suggests that these children may be helped by early cognitive-behavioral intervention. PMID- 18412061 TI - The role of neuroimaging for the diagnosis and treatment of anxiety disorders. AB - Neuroimaging comprises a set of tools, which include different types of magnetic resonance imaging such as functional magnetic resonance imaging (fMRI), Magnetic Resonance Spectroscopy (MRS), Arterial Spin Labeling (ASL), and radiotracer imaging such as positron emission tomography and single photon emission computed tomography. The focus of this review is to address the question whether fMRI can contribute to the diagnosis and treatment of anxiety disorders. Key anxiety processes and neural substrates are reviewed. The main findings and shortcomings of fMRI in the context of anxiety are briefly summarized. Finally, the next stages of developing fMRI for diagnosis and treatment are highlighted. The main conclusion of this review is that fMRI could become a clinical tool for the diagnosis and treatment of anxiety disorders, but neuroimaging groups will need to better develop its specificity and sensitivity so that fMRI results can be meaningful for an individual patient not just for groups of individuals. PMID- 18412063 TI - Genetics of anxiety: would the genome recognize the DSM? AB - The nosology of anxiety disorders has undergone substantial evolution over the past several decades. The modern classification of these disorders dates to the publication of Diagnostic and Statistical Manual-III (DSM-III) in 1980, but the validity of the current diagnostic categories has been the subject of controversy. Genetic research can help clarify the boundaries of diagnostic categories by examining the etiologic relationships among them. The question posed in the title of this article asks to what degree the DSM-IV definitions of the anxiety disorders are supported by the evolving body of research on the genetic basis of pathologic anxiety. With DSM-V on the horizon, there is a renewed imperative to examine the structure of these disorders. In this article, we address this issue by, first, providing a brief update about the current status of genetic research on anxiety disorders and then considering whether the evidence suggests that genetic influences conform to or transcend DSM definitions. Finally, we discuss future directions for the genetic dissection of anxiety disorders. PMID- 18412064 TI - The Carotid Artery Revascularization and Endarterectomy (CARE) registry: objectives, design, and implications. AB - This is the first comprehensive national registry that will provide data characterizing contemporary results of carotid endarterectomy (CEA) and carotid artery stenting (CAS). Carotid endarterectomy (CEA) has become the standard revascularization therapy to prevent stroke in patients with carotid artery disease, while carotid artery stenting (CAS) offers a percutaneous alternative in selected patients. Given the rapid growth in the numbers of CAS procedures being performed, there is a critical need for a national program to assess quality outcomes. The Carotid Artery Revascularization and Endarterectomy (CARE) Registry was developed through a multispecialty collaboration resulting in a comprehensive data collection tool for carotid revascularization procedures. The intent of the CARE registry is to collect and analyze clinical data to measure clinical practice, patient outcomes, and enable quality improvement for carotid revascularization. Finally, the CARE Registry satisfies the Center for Medicare and Medicaid Services (CMS) data reporting criteria for reimbursement. PMID- 18412065 TI - Novel use of ultrasound guidance for recanalization of iliac, femoral, and popliteal arteries. AB - OBJECTIVES: We attempted to demonstrate the safety and feasibility of intravascular ultrasound (IVUS)-guided endluminal endovascular therapy for chronic total occlusion (CTO) of the iliac and/or femoropopliteal arteries using Treasure 12, a stiff guidewire for peripheral CTO of 0.018 in. in monodirectional approach setting. BACKGROUND: Endovascular therapy for CTO of the iliac and femoropopliteal arteries is still technically challenging. METHODS: Endovascular therapy was performed in 110 patients who were admitted to Hyogo College of Medicine Hospital with iliac and/or femoropopliteal artery disease from January 2006 to October 2007. We retrospectively analyzed the data of consecutive 47 patients who underwent the endovascular therapy by monodirectional approach for CTO lesions of the iliac and femoropopliteal arteries. From January 2006 to November 2006 (phase 1), standard technique was applied for the treatment of 21 lesions. From December 2006 to October 2007 (phase 2), IVUS-guided endovascular therapy was applied for the treatment of 31 lesions. RESULTS: Clinical and lesion characteristics in phase 1 were not significantly different from those in phase 2. The overall initial technical success rate improved from 81% in phase 1 to 97% in phase 2. There were no significant differences in radiation exposure time between phase 1 and phase 2. Total volume of contrast material was significantly smaller in phase 2 than in phase 1 (P < 0.01). CONCLUSIONS: The IVUS-guided endovascular therapy for CTO using Treasure 12 guidewire is feasible and safe, minimizes contrast material, and has a high initial technical success rate. PMID- 18412066 TI - Safety and effectiveness of drug eluting stent in patients with ST elevation myocardial infarction undergoing primary angioplasty. AB - BACKGROUND: Drug eluting stents (DES) have recently been proven to further reduce restenosis and revascularization rate in comparison to bare metal stents in elective procedures. Most early DES trials did not include patients undergoing primary percutaneous coronary intervention (PCI) for ST-segment elevation MI, because these patients tend to have lower restenosis rates than other patient groups and delayed endothelization of these stents raises concern about a possible increase of thrombotic complications in the setting of STEMI. AIM: To confirm the safety and effectiveness of DES in patients with STEMI in a real world scenario. METHODS: From January 2004 to December 2006, clinical and angiographic data of 370 patients with STEMI treated with primary PCI have been analyzed. Patients were retrospectively followed for the occurrence of major adverse cardiac events (MACE): death, reinfarction and target vessel revascularization (TVR). RESULTS: Overall, 120 patients received DES (32%, DES group) and 250 received bare metal stents (68%, BMS group) in the infarct related artery. Compared with the BMS group, DES patients were younger, (mean age 56 +/- 12 vs. 65 +/- 10; P < 0.001) had more often diabetes mellitus (47% vs. 14% P < 0.001), anterior localization (65% vs. 45%; P < 0.0011) and less cardiogenic shock at admission (4% vs. 7%; P < 0.001). The angiographic characteristics in the DES group showed longer lesions (23 mm vs. 19 mm) and smaller diameter of vessels (2.5 mm vs. 3.0 mm). After a median follow-up of 24 +/- 9 months, there was no significant difference in the rate of stent thrombosis (1.6% in the DES group vs. 1.2% in the BMS group, P = ns). The incidence of MACE was significantly lower in the DES group compared with the BMS group (HR 0.56 [95% CI: 0.3-0.8]; P = 0.01), principally due to the lower rate of TVR (HR 0.41 [95% CI: 0.2-0.85]; P = 0.01). CONCLUSIONS: Utilization of DES in the setting of primary PCI for STEMI, in our "real world," was safe and improved the 3-year clinical outcome compared with BMS reducing the need of TVR. PMID- 18412067 TI - Don't you care: you should. PMID- 18412068 TI - Recanalization of chronic peripheral artery occlusions: moving forward by looking sideways. PMID- 18412069 TI - Common femoral artery stenosis after deployment of vascular clip closure device. AB - We describe a case of symptomatic common femoral artery stenosis following use of a vascular clip closure device (StarClose). Operative repair was performed, with removal of the clip device and subsequent vein patch angioplasty. PMID- 18412070 TI - The road to perfect closure--still unpaved. PMID- 18412071 TI - Distal embolization during intracranial carotid artery stenting. AB - Intracranial stenting, with or without balloon angioplasty, has emerged as an effective alternative to medical therapy for intracranial atherosclerosis. However, this strategy appears to involve the potential risk of distal embolization, which may lead to disability and death. Here, the authors have described a successful case of distal balloon-protected intracranial carotid artery stenting for clinically and angiographically unstable intracarotid artery stenosis revealing aspirated embolic materials containing yellow plaque, red thrombus, and white debris. PMID- 18412072 TI - In search of perfect closure. PMID- 18412073 TI - Cutaneous migration of the StarClose device. AB - We report the first case of a transcutaneous migration of a vascular clip closure device 10 weeks after placement during femoral artery puncture for percutaneous transluminal angioplasty. We postulate focal inflammatory vascular wall changes as the predisposing factor in this patient with giant cell arteritis. PMID- 18412074 TI - Safety and efficacy of drug-eluting stent for ST-segment elevation myocardial infarction in an unselected consecutive cohort. AB - OBJECTIVE: The objective of this study is to investigate the clinical outcome of a large cohort of patients with ST-segment elevation myocardial infarction (STEMI) treated with drug-eluting stents (DES) compared to bare metal stents (BMS). BACKGROUND: Several randomized controlled trials have demonstrated that PCI with the routine use of DES is safe and effective in patients with STEMI. However as randomized trials have strict inclusion criteria, further studies in unselected patient populations are needed. METHODS: We performed a retrospective cohort analysis of the Mayo Clinic PCI database. A total of 552 consecutive STEMI patients who underwent a DES implantation between May 2003 and April 2006 were included in the study and compared to 557 who had BMS for STEMI earlier. No specific patient subsets were excluded. RESULTS: Procedural success was achieved in 532 patients (96%). During initial hospitalization, 16 patients (2.9%) died and 8 (1.5%) suffered from a recurrent myocardial infarction. The median follow up was 23 months (IQR: 13-27 months). At 12 months post discharge, the rate of target lesion revascularization and death were 2.9% and 3.7%, respectively, and survival free of major adverse cardiac events (MACE) was 90.9%. These rates were similar to or lower than those of patients treated for STEMI with BMS prior to the availability of DES. CONCLUSION: DES are safe and effective in the treatment of STEMI in an unselected cohort; 90.9% of patients are free of MACE at 12 months post discharge. PMID- 18412075 TI - Drug eluting stents in AMI: keep the data coming. PMID- 18412076 TI - A novel use of a split sheath to maintain vessel access when guidewire exchange is not possible. AB - After inserting a catheter into a vessel, guidewire exchange technique must be used to maintain vascular access. If the lumen of the catheter is blocked and the guidewire exchange cannot be used, the catheter can only be removed, vascular access is lost, and an alternate vascular approach attempted. We describe a technique to remove the blocked catheter and still maintain vascular access. PMID- 18412077 TI - Physical aspects of excimer laser angioplasty for undilatable lesions. PMID- 18412078 TI - How to show the evidence. PMID- 18412079 TI - What anti-thrombotic therapy is best with primary PCI for acute ST elevation myocardial infarction: how should the HORIZONS trial change current practice? AB - The current standard of care for anti-thrombotic therapy with primary PCI for acute ST elevation myocardial infarction (STEMI) is aspirin, clopidogrel, unfractionated heparin and platelet glycoprotein IIb/IIIa inhibitors. However, heparin and glycoprotein IIb/IIIa inhibitors are associated with a high incidence of bleeding, and many of the trials documenting benefit with this therapy were performed before the widespread use of stents and clopidogrel. Bivalirudin is a direct thrombin inhibitor which has been found to have similar efficacy with less bleeding compared with heparin plus glycoprotein IIb/IIIa inhibitors when used with elective PCI and with PCI for unstable angina and non-ST elevation myocardial infarction. The HORIZONS trial evaluated bivalirudin compared with unfractionated heparin and IIb/IIIa inhibitors in patients with STEMI treated with primary PCI and found similar MACE (major adverse cardiac events) with less bleeding and a lower incidence of net adverse clinical events (MACE or major bleeding) at 30 days. Mortality at 30 days was also significantly less with bivalirudin. These results make a strong case for the use of bivalirudin with primary PCI in the great majority of patients with STEMI, with the possible exception of patients with cardiogenic shock or stent thrombosis, and patients with a large thrombus burden or no re-flow following PCI. In the latter case, platelet glycoprotein IIb/IIIa inhibitors would be used as a bail-out strategy. PMID- 18412080 TI - Another addition to the tool box. PMID- 18412081 TI - Stent implantation is effective treatment of vascular stenosis in young infants with congenital heart disease: acute implantation and long-term follow-up results. AB - BACKGROUND: Stents implantation in infants has been shown to be feasible, however, there are no published reports examining long-term outcomes. Concerns exist regarding creation of fixed obstructions secondary to the stent if expansion to larger diameters over time is not possible. METHODS: A retrospective analysis of the earliest consecutive series of infants who underwent stent placement at our institution between October 1995 and December 1999. RESULTS: Implantation of 33 stents were attempted in 27 infants, median age = 10 (25-24) months, wt = 8.1 (3.4-14.5) kg. Stents used were as follows: 16 large, 13 medium, and 4 coronary. Acute implant success was 94%. There were three nonprocedure related deaths within 30 days of implantation, 1 patient was lost to follow-up and 1 had acute stent thrombosis. The remaining 22 patients (26 stents) form the long-term follow-up study group. Nineteen stents underwent 33 redilations. Following latest redilation, 67.0 (37-113) months postimplantation, minimal luminal diameter increased from 7.0 +/- 1.8 mm immediately following implantation to 8.7 +/- 2.3 mm (P < 0.001). Seven stents were electively removed/ligated during a planned surgery. All 18 remaining in situ stents are patent without significant obstruction 102 (84-116) months following implantation. There was one late death 51 months after stent implantation. The remaining 21 patients are alive and well. CONCLUSIONS: Stent implantation in infants is safe and effective. Serial redilation is possible to keep pace with somatic growth; however, efforts should be made to implant stents with adult diameter potential in children who will not require further cardiac surgery. Implantation of small- and medium-sized stents can provide effective palliation and should be considered in carefully selected infants who will ultimately require future surgery. PMID- 18412082 TI - Percutaneous management of cyanosis in Fontan patients using Amplatzer occluders. AB - OBJECTIVES: To determine causes of cyanosis and to evaluate percutaneous management of cyanosis in a group of consecutive Fontan patients. BACKGROUND: A variety of communications allow a right-to-left shunt in Fontan circulation causing cyanosis and these communications are amenable to percutaneous closure. METHODS: Between November 1997 and November 2007, 45 consecutive patients ranging in age from 2.5 to 26 years (median 8 years) with Fontan circulation and cyanosis underwent cardiac catheterization and percutaneous closure of superfluous communications using different types of Amplatzer occluders. RESULTS: Altogether, 51 communications were detected and 50 of them were closed. In 5 patients (11.1%), multiple communications were present and were closed. Fenestration was detected in 37 of 41 patients (90.2%) after total cavopulmonary connection and all were closed by Amplatzer septal occluders. Five venous collaterals were revealed in 3 of 41 patients (7.3%) with total cavopulmonary anastomosis and were closed by Amplatzer vascular plugs. In 3 patients, lateral tunnel leaks were detected and were closed by Amplatzer PFO occluders. In 4 patients after Kawashima operation, 3 major pulmonary arterio-venous malformations and single venous collateral contributed to the cyanosis and all were closed using Amplatzer vascular plugs. CONCLUSIONS: Different communications cause cyanosis in Fontan patients. Progressive decline of percutaneous oxygen saturation is suggestive of development of venous collaterals or pulmonary arteriovenous malformations. Superfluous communications are amenable to percutaneous closure using various types of Amplatzer occluders. A novel use of an Amplatzer PFO occluder for the percutaneous closure of a lateral tunnel leak is described. PMID- 18412083 TI - Stent implantation as effective treatment of vascular stenosis in young infants with congenital heart disease: acute implantation and follow-up. PMID- 18412084 TI - Novel catheter positioning technique for atretic pulmonary valve perforation. AB - Percutaneous pulmonary valve perforation in selected neonates with pulmonary valve atresia and intact ventricular septum has been well established as a therapeutic option. Pulmonary valve perforation is associated with significant risks including perforation of the right ventricle. Optimal catheter positioning is crucial to the success of the procedure. This report describes a novel technique for accurate positioning of a guiding catheter during perforation of an atretic pulmonary valve. PMID- 18412085 TI - A new device to close secundum atrial septal defects: first clinical use to close multiple defects in a child. AB - Transcatheter device closure of secundum atrial septal defects (ASD) has become an accepted treatment modality. Currently, the only device that can be used for large defects is the Amplatzer Septal Occluder. We report on a new device (Occlutech Figulla), to close multiple ASDs in a child with two large defects using 12 and 15 mm devices. PMID- 18412086 TI - A statement on ethics from the HEART group. PMID- 18412088 TI - Commentary for "Comparison of drug eluting stents with bare metal stents in unselected patients with acute myocardial infarction". PMID- 18412090 TI - Another "nail in the coffin" for protection devices in acute MI? PMID- 18412091 TI - Recanalization of chronically occluded coronaries: a step forward in understanding its impact. PMID- 18412092 TI - Innovations in bifurcations. PMID- 18412093 TI - Transvaginal sonographic measurement of cervical length to predict preterm birth in asymptomatic women at increased risk: a systematic review. AB - OBJECTIVES: To estimate the ability of cervical length measured by transvaginal ultrasonography in asymptomatic high-risk women to predict spontaneous preterm birth. METHODS: MEDLINE, PubMed, EMBASE and the Cochrane Library were searched for articles published in any language between January 1980 and July 2006, using the keywords 'transvaginal ultrasonography' or ('cervix' and ('ultrasound' or 'ultrasonography' or 'sonography')); and ('preterm' or 'premature') and ('delivery' or 'labour/labor' or 'birth'), identifying cohort studies evaluating transvaginal ultrasonographic cervical length measurement in predicting preterm birth in asymptomatic women who were considered at increased risk (because of a history of spontaneous preterm birth, uterine anomalies or excisional cervical procedures), with intact membranes and singleton gestations. The primary analysis included all studies meeting the inclusion criteria. Secondary analyses were also performed specifically for (1) women with a history of spontaneous preterm birth; (2) those who had undergone an excisional cervical procedure; and (3) those with uterine anomalies. RESULTS: Fourteen of 322 articles identified (involving 2258 women) met the criteria for systematic review. Cervical length measured by transvaginal ultrasonography predicted spontaneous preterm birth. The shorter the cervical length cut-off the higher the positive likelihood ratio (LR). The most common cervical length cut-off was < 25 mm. Using this cut-off to predict spontaneous preterm birth at < 35 weeks, transvaginal ultrasonography at < 20 weeks' gestation revealed LR+ = 4.31 (95% CI, 3.08-6.01); at 20-24 weeks, LR+ = 2.78 (95% CI, 2.22-3.49); and at > 24 weeks, LR+ = 4.01 (95% CI, 2.53-6.34). In women with a history of spontaneous preterm birth (six studies involving 663 women) cervical length at < 20 weeks revealed LR+ = 11.30 (95% CI, 3.59-35.57) and at 20-24 weeks LR+ = 2.86 (95% CI, 2.12-3.87), but there were limited data on the use of cervical length of more than 24 weeks in this group (one study involving 42 women). In women who had had excisional cervical procedures, two studies presented data on cervical length (one at < 24 weeks and one at > 24 weeks), finding cervical length at < 24 weeks to be predictive of spontaneous preterm birth at < 35 weeks (LR+ = 2.91, 95% CI, 1.69-5.01). One study (of 64 women) evaluated cervical length in women with uterine anomalies, finding it predictive of spontaneous preterm birth at < 35 weeks (LR+ = 8.14, 95% CI, 3.12 21.25). CONCLUSION: Cervical length measured by transvaginal ultrasonography in asymptomatic high-risk women predicts spontaneous preterm birth at < 35 weeks. Further research may be warranted to evaluate the use of transvaginal ultrasonography after 24 weeks' gestation in women with a history of spontaneous preterm birth, and in women with uterine anomalies. PMID- 18412094 TI - Multi-class, multi-residue liquid chromatography/tandem mass spectrometry screening and confirmation methods for drug residues in milk. AB - This paper describes the development and optimization of a multi-residue veterinary drug screening method for whole milk. The drug residues of regulatory interest in milk include beta-lactams, sulfonamides, tetracyclines, fluoroquinolones, and macrolides. Milk samples were extracted with acetonitrile and the samples were then subjected to a clean-up procedure using a bonded solid phase extraction cartridge and a molecular weight cut-off filter. Liquid chromatography/tandem mass spectrometry (LC/MS/MS) triple quadrupole electrospray methods were developed to monitor for the drugs in milk. Since established tolerance levels are set for most of these drugs in milk, the initial screening procedure was semi-quantitative, where samples were compared to the response of a positive control. The positive control, consisting of an extract from a portion of milk fortified with the drugs at half their allowed levels, was used to set the laboratory's minimum response criteria for unknown samples. Confirmatory analyses, with additional ion transitions for each residue, were performed on the same extracts. PMID- 18412095 TI - Sonographic characteristics of the uterine cavity following first-trimester uterine evacuation. AB - OBJECTIVE: To characterize the sonographic appearance of the uterine cavity after first-trimester uterine evacuation and to follow the evolution of these findings in an attempt to reduce the number of unnecessary surgical interventions following evacuation. METHODS: We studied retrospectively the sonographic characteristics of the uterine cavity in 599 women, 5-8 days after first trimester uterine evacuation. The patients were grouped according to the sonographic appearance of the uterine cavity. Patients with abnormal sonographic patterns were followed weekly until sonographic resolution. RESULTS: Group 1 included 351 (58.6%) women with normal endometrium; Group 2 included 130 (21.7%) women with hypoechoic endometrial content only; Group 3 included 69 (11.5%) patients with mixed type (hypoechoic and hyperechoic) endometrial content; Group 4 included 49 (8.2%) patients with hyperechoic endometrial content only. The time needed until the uterine cavity was considered normal was significantly longer in Group 4 (median, 12 days) compared with Groups 3 and 2 (8 and 9 days, respectively, P < 0.0001). The duration of vaginal bleeding after the surgical procedure was longer in Group 4 (median, 10 days) compared with Groups 3, 2 and 1 (9, 7 and 5 days, respectively, P < 0.0001). Clinically, the patients were divided into two groups: asymptomatic (575 patients) and symptomatic (24 patients). The thickness of the abnormal endometrial content in the asymptomatic patients gradually decreased until normalization, around the time of menstruation. There was no such change in the symptomatic patients, who eventually needed surgical intervention. CONCLUSIONS: An abnormal intrauterine sonographic pattern 5-8 days following first-trimester uterine evacuation is common and usually resolves spontaneously around menses. Therefore, in asymptomatic patients, we recommend a conservative approach. PMID- 18412096 TI - PET-based investigation of cerebral activation following intranasal trigeminal stimulation. AB - The present study aimed to investigate cerebral activation following intranasal trigeminal chemosensory stimulation using O15-H2O-PET. A total of 12 healthy male participants underwent a PET scan presented with four scanning conditions; two left-sided intranasal CO(2)-stimuli and two matched baseline conditions consisting of odorless air. CO(2) was used as it produces burning and stinging sensations. Stimulation started 20 s before intravenous injection of the isotope and lasted for the first 60 s of the 5 min scan time. A comparison between CO(2) and baseline showed a pronounced activation of the trigeminal projection area at the base of the postcentral gyrus (primary and secondary somatosensory cortex) which was more intense for the right hemisphere, contralateral to the side of stimulation. In addition, activation was also found in the piriform cortex which is typically activated following odor presentation and thus thought of as primary olfactory cortex. In conclusion, and in line with previously published work, our data suggest that intranasal trigeminal stimulation not only activates somatosensory projection areas, but that it also leads to activation in cerebral areas associated with the processing of olfactory information. This may be interpreted in terms of the intimate relation between the intranasal chemosensory systems. PMID- 18412097 TI - Ectopic testicular xenografts from newborn hamsters (Phodopus sungorus) show better spermatogenic activity in aged compared with young recipients. AB - The mechanisms behind testicular aging are poorly understood. Previous studies suggest that the testicular microenvironment is more affected by age than the male germ cell lineage. Here we analyze male reproductive aging using a unique xenografting approach. By exposing young and aged mice to newborn hamster testicular tissue, we can explore (a) whether the development and endocrine activity of hamster testicular grafts and the initiation of stem cell activity within them are affected by age of the recipients and (b) whether the endocrine response to the xenografted hamster tissue varies with recipient age. Newborn Djungarian hamster (Phodopus sungorus) testes were grafted into young (12 weeks) and aged (1 year) adult castrated nude mice. We also analyzed intact and castrated young and old control groups. After 13 weeks, 100 grafts were recovered from a total of 15 recipients and were histologically analyzed. Anatomical and endocrine parameters were recorded for each recipient as well as for the controls. Xenografted recipients responded with a normalization of their endocrine and anatomical parameters to an extent typical for their age. Although recipient age did not significantly affect graft survival and size, histopathological changes as well as spermatogenic damage within the grafts were more pronounced in the young recipients (56% Sertoli-cell-only tubules vs. 32% in the old recipients). We conclude from our data that the androgen-related changes associated with male reproductive aging are not primarily controlled by the testis. We speculate that the better development of testicular grafts in aged recipients may be owing to immunosenescence. PMID- 18412098 TI - Genetics of post-traumatic stress disorder: informing clinical conceptualizations and promoting future research. AB - The purpose of this article is to provide an overview of genetic research involving post-traumatic stress disorder (PTSD). First, we summarize evidence for genetic influences on PTSD from family investigations. Second, we discuss the distinct contributions to our understanding of the genetics of PTSD permitted by twin studies. Finally, we summarize findings from molecular genetic studies, which have the potential to inform our understanding of underlying biological mechanisms for the development of PTSD. PMID- 18412099 TI - The genetics of obsessive compulsive disorder: a review of the evidence. AB - Obsessive compulsive disorder (OCD) is a common psychiatric disorder that can have disabling effects on both adults and children. Twin, family, segregation, and linkage studies have demonstrated that OCD is familial, that the familiality is due in part to genetic factors and there are regions of the genome which very likely harbor susceptibility loci for OCD. Over 60 candidate gene studies have been conducted. Most studies have focused on genes in the serotonergic and dopaminergic pathways. Unfortunately, none have achieved genome-wide significance and with the exception of the glutamate transporter gene, none have been reliably replicated. Future research will requite much larger samples and the collaboration of researchers to be able to identify susceptibility loci for OCD. PMID- 18412100 TI - Genetics of anxiety disorders: complexities and opportunities. PMID- 18412101 TI - What is the genetic relationship between anxiety and depression? AB - Anxiety and depression share a long, close history in psychiatric nosology and treatment. The anxiety disorders, individually and as a group, exhibit remarkably high rates of comorbidity with each other and with major depression. Analyses done in large-scale epidemiologic surveys have identified major patterns of phenomenological overlap between these conditions. Researchers have tested hypotheses of shared genetic etiologies as a potential basis of this relationship. In general, available family studies have found mixed evidence for co-aggregation of anxiety and depressive disorders, while twin studies more definitively indicate that shared genetic risk factors largely account for this comorbidity. Some of this appears to be accounted for by genetic variation in personality traits that broadly predispose to anxiety and depression. Molecular genetic studies of these conditions, though too early to draw firm conclusions, thus far provide tentative support for specific genetic loci that may generally influence susceptibility across the anxiety-depressive spectrum. PMID- 18412102 TI - Recent advances in the neurobiology of anxiety disorders: implications for novel therapeutics. AB - Anxiety disorders are a highly prevalent and disabling class of psychiatric disorders. This review focuses on new directions in neurobiological research and implications for the development of novel psychopharmacological treatments. Neuroanatomical and neuroimaging research in anxiety disorders has centered on the role of the amygdala, reciprocal connections between the amygdala and the prefrontal cortex, and, most recently, alterations in interoceptive processing by the anterior insula. Anxiety disorders are characterized by alterations in a diverse range of neurochemical systems, suggesting ample novel targets for drug therapies. Corticotropin-releasing factor (CRF) concentrations are elevated in a subset of anxiety disorders, which suggests the potential utility of CRF receptor antagonists. Pharmacological blockade of the memory-enhancing effects of stress hormones such as glucocorticoids and noradrenaline holds promise as a preventative approach for trauma-related anxiety. The glutamatergic system has been largely overlooked as a potential pharmacological target, although convergent preclinical, neuroimaging, and early clinical findings suggest that glutamate receptor antagonists may have potent anxiolytic effects. Glutamatergic receptor agonists (e.g., D-cycloserine) also have an emerging role in the treatment of anxiety as facilitators of fear extinction during concurrent behavioral interventions. The neuropeptides substance P, neuropeptide Y, oxytocin, orexin, and galanin are each implicated in anxiety pathways, and neuropeptide analogs or antagonists show early promise as anxiolytics in preclinical and/or clinical research. Each of these active areas of research holds promise for expanding and improving evidence-based treatment options for individuals suffering with clinical anxiety. PMID- 18412104 TI - Reintegrative Shaming Theory, moral emotions and bullying. AB - This article investigates the usefulness of Reintegrative Shaming Theory (RST) in explaining the bullying of siblings in families and peers in schools. Questionnaires were completed by 182 children aged 11-12 years in ten primary schools in Nicosia, Cyprus, about sibling and peer bullying. A vignette-based methodology was used to investigate children's expectations of the type of shaming their parents would offer in response to their possible wrong doing. Children were also asked questions about the emotions they would have felt (i.e. shame, remorse, guilt or anger) if they were in the position of the child in the vignette. The level of bonding toward each parent was also examined. In agreement with the theory, a path analysis showed that mother bonding influenced children's expectations of the type of shaming offered by parents. Disintegrative shaming (i.e. shaming offered in a stigmatizing or rejecting way) had a direct effect on the way children managed their shame. Shame management directly influenced sibling and peer bullying. Father bonding had no direct or indirect effects in the model. Against the theory, reintegrative shaming (i.e. shaming offered in the context of approving the wrongdoer while rejecting the wrongdoing) did not have a direct effect on shame management. Beyond the postulates of RST, mother bonding-a plausible indicator of family functioning-had a direct effect on sibling and peer bullying. Mother bonding had a stronger effect for boys than for girls. It is concluded that RST is useful in explaining the link between family factors and bullying, and that RST has cross-cultural applicability. PMID- 18412103 TI - Identification of neurogenetic pathways of risk for psychopathology. AB - Imaging genetics has been a highly effective and increasingly applied strategy for identifying the impact of genetic polymorphisms on individual differences in neural circuitry supporting complex behaviors. The application of imaging genetics towards further elucidating neural circuitry associated with the pathophysiology of psychiatric illness is of particular interest given its potential to guide the development and improvement of current therapeutic methods. The identification of genetic variants that contribute to or predict the disruption of specific neural pathways associated with psychopathology may also serve as useful markers of risk demarcating individuals with elevated susceptibility for psychiatric illness and affording early or even preemptive treatment strategies. In the continued development of this technique, recent multimodal neuroimaging strategies and studies examining the effects of multiple genes in concert within large subject populations have shown promise in the development of a more complete understanding of the interrelationships between genes, brain function, behavior and associated risk for psychopathology. PMID- 18412105 TI - Effects of cuminaldehyde on melanoma cells. AB - Cuminaldehyde (4-isopropylbenzaldehyde) suppressed melanin formation in cultured murine B16-F10 melanoma cells in a dose-dependent decrease up to 0.25 mm without affecting cell growth. Approximately 30% suppression in melanin production resulted when the cells were cultured with 0.25 mm of cuminaldehyde. This activity was not noticeable with cultured human A375 melanoma cells. PMID- 18412106 TI - In vivo MRI assessment of a novel GdIII-based contrast agent designed for high magnetic field applications. AB - Gd(3)L is a trinuclear Gd(3+) complex of intermediate size, designed for contrast agent applications in high field magnetic resonance imaging (H(12)L is based on a trimethylbenzene core bearing three methylene-diethylenetriamine- N,N,N'',N'' tetraacetate moieties). Thanks to its appropriate size, the presence of two inner sphere water molecules and a fast water exchange, Gd(3)L has remarkable proton relaxivities at high magnetic field (r(1) = 10.2 vs 3.0 mM(-1) s(-1) for GdDOTA at 9.4 T, 37 degrees C, in H(2)O). Here we report an in vivo MRI feasibility study, complemented with dynamic gamma scintigraphic imaging and biodistribution experiments using the (153)Sm-enriched analog. MRI experiments were performed at 9.4 T in mice with Gd(3)L and the commercial contrast agent gadolinium(III) 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetate (GdDOTA). Gd(3)L was well tolerated by the animals at the dose of 8 micromol Gd kg(-1) body weight. Dynamic contrast enhanced (DCE) images showed considerably higher signal enhancement in the kidney medulla and cortex after Gd(3)L injection than after GdDOTA injection at an identical dose. The relaxation rates, DeltaR(1), were calculated from the IR TrueFISP data. During the excretory phase, the DeltaR(1) for various tissues was similar for Gd(3)L and GdDOTA, when the latter was injected at a three-fold higher dose (24 vs 8 micromol Gd kg(-1) body weight). These results point to an approximately three times higher in vivo relaxivity (per Gd) for Gd(3)L relative to GdDOTA, thus the ratio of the relaxivities of the two compounds determined in vitro is retained under in vivo conditions. They also indicate that the two inner sphere water molecules per Gd in Gd(3)L are not substantially replaced by endogenous anions or other donor groups under physiological conditions. Gd(3)L has a pharmacokinetics typical of small, hydrophilic complexes, involving fast renal clearance and no retention in the blood pool. The dynamic gamma scintigraphic studies and the biodistribution experiments performed in Wistar rats with (153)Sm-enriched (*)Sm(3)L are also indicative of a fast elimination via the kidneys. PMID- 18412107 TI - Bottlenecks in molecular testing for rare genetic diseases. AB - Despite the impressive progress in our understanding of the genetic causes of genetic diseases over the past decade, molecular diagnosis for rare genetic disorders is still in its infancy, being slow, expensive, unreliable, insufficient, and ill-organized in many countries. This leaves the gap between the hype of the current genomic research and the hope for a simple genetic diagnosis too large for patients and families affected with genetic disease. The bottlenecks in the molecular testing for rare genetic disorders are discussed below. PMID- 18412108 TI - The genetic basis of panic and phobic anxiety disorders. AB - Panic disorder and phobic anxiety disorders are common disorders that are often chronic and disabling. Genetic epidemiologic studies have documented that these disorders are familial and moderately heritable. Linkage studies have implicated several chromosomal regions that may harbor susceptibility genes; however, candidate gene association studies have not established a role for any specific loci to date. Increasing evidence from family and genetic studies suggests that genes underlying these disorders overlap and transcend diagnostic boundaries. Heritable forms of anxious temperament, anxiety-related personality traits and neuroimaging assays of fear circuitry may represent intermediate phenotypes that predispose to panic and phobic disorders. The identification of specific susceptibility variants will likely require much larger sample sizes and the integration of insights from genetic analyses of animal models and intermediate phenotypes. PMID- 18412109 TI - Inactivation of the CDKL3 gene at 5q31.1 by a balanced t(X;5) translocation associated with nonspecific mild mental retardation. AB - We have investigated the breakpoints of a balanced reciprocal translocation between chromosomes X and 5, [46,X,t(X;5)(p11.1;q31.1)], in a woman with mild mental retardation (MR). Methylation studies showed a 100% skewed X-inactivation in patient-derived lymphocytes. Cloning and sequencing of the junction fragment from the X derivative showed that the breakpoint occurred in intron 3 of the CDKL3 gene on chromosome 5 and in a region devoid of genes on chromosome X. Quantitative RT-PCR analyses on patient-derived lymphoblastoid cells documented a significant 50% decrease of the CDKL3 transcript level. Allelic expression analysis, using an intronic SNP that was RT-PCR amplified from CDKL3 pre-mRNA, provided further evidence that the CDKL3 gene was transcribed from only one allele. Decreased CDKL3 gene expression was definitively confirmed at the protein level by immunoblot analysis. CDKL3 is a member of a subset of the cdc2-related protein kinase family that shows similarity to both mitogen-activated protein kinases (MAPK) and cyclin-dependant kinases (cdks). Importantly, one member of the family, CDKL5, has been implicated in atypical Rett syndrome, West syndrome, and X-linked infantile spasm, all including MR as a manifestation. Expression studies demonstrated that the mouse homologue, mCdkl3, was expressed in all brain regions investigated and throughout mouse development, a pattern that is consistent with a role in development and brain function. Together the data suggest that haploinsufficiency of CDKL3 in the t(X;5) patient contributes to her phenotype, and that the CDKL3 gene is a strong candidate for nonsyndromal autosomal dominant MR. PMID- 18412110 TI - Dominantly inherited lop ears. PMID- 18412111 TI - Partial Xp11.23-p11.4 duplication with random X inactivation: clinical report and molecular cytogenetic characterization. AB - Partial duplications of the short arm of the X chromosome are relatively rare and have been described in males and females. We describe a 4 10/12-year-old girl presenting with developmental delay, severe language retardation and minor anomalies with slightly elevated head circumference (+1.8 SD), prominent forehead, wide palpebral fissures and anteverted nares. No pigmentary dysplasia of the skin was present. The external genitalia were normal. The karyotype completed by cytogenetic analysis with the Whole Chromosome Painting probe of chromosome X revealed a de novo partial duplication of the short arm of an X chromosome. In order to further characterize the duplicated segment, we used a series of BAC probes extending from band Xp11.22 to Xp22.1. BACs from Xp11.23 to Xp11.4 were duplicated. The karyotype was finally defined as 46,X,dup(X)(p11p11).ish dup(X)(p11.23p11.4)(WCPX+,RP11-416I6++,RP11-386N14++,RP11 466C12++). The X-inactivation status was studied using the human androgen receptor (HUMARA) and the FRAXA locus methylation assay. Unexpectedly, the two X chromosomes were found to be randomly inactivated, in the proband. Indeed, usually, in women with structurally abnormal X chromosome, the abnormal X chromosome is preferentially inactivated and those patients share an apparent normal phenotype. So, we speculate that in the present case, the phenotype of the patient could be explained by a functional disomy of the genes present in the duplicated region. We will discuss the possible implication of these genes on the observed phenotype. PMID- 18412112 TI - The neural basis of response inhibition and attention allocation as mediated by gestational age. AB - Children and adolescents born before 33 weeks of gestation, that is very preterm, may experience problems with the inhibitory control of behaviour and the allocation of attention. Previous functional magnetic resonance imaging (fMRI) studies have found preterm-born adolescents to display altered brain activation in tasks measuring inhibitory control. However, adolescence is a period during which dynamic changes are occurring in the brain, and it is not yet known whether these functional alterations will persist into adulthood, or instead reflect developmental delay. This study used an event-related fMRI Go/No-Go motor response inhibition paradigm, which included an oddball task measuring attention allocation to infrequent stimuli, to compare blood-oxygen-level-dependent (BOLD) signal between 26 preterm-born adults and 21 controls. Group differences in brain activation were observed in inhibition and attention networks during both conditions. During motor response inhibition, preterm-born participants compared to controls showed increased BOLD signal in medial and right lateral posterior brain regions, including middle temporal/occipital gyrus, posterior cingulate gyrus and precuneus. During oddball trials, preterm-born young adults displayed attenuated brain activation in a fronto-parietal-cerebellar network which is involved in mediating attention allocation. This pattern of reduced brain activation in task-relevant regions of attention allocation, and increased activation in posterior brain regions during inhibitory control, suggests adult alteration of inhibition and attention processing following very preterm birth, which may reflect a developmental delay. PMID- 18412113 TI - An fMRI normative database for connectivity networks using one-class support vector machines. AB - The application of functional magnetic resonance imaging (fMRI) in neuroscience studies has increased enormously in the last decade. Although primarily used to map brain regions activated by specific stimuli, many studies have shown that fMRI can also be useful in identifying interactions between brain regions (functional and effective connectivity). Despite the widespread use of fMRI as a research tool, clinical applications of brain connectivity as studied by fMRI are not well established. One possible explanation is the lack of normal patterns and intersubject variability-two variables that are still largely uncharacterized in most patient populations of interest. In the current study, we combine the identification of functional connectivity networks extracted by using Spearman partial correlation with the use of a one-class support vector machine in order construct a normative database. An application of this approach is illustrated using an fMRI dataset of 43 healthy subjects performing a visual working memory task. In addition, the relationships between the results obtained and behavioral data are explored. PMID- 18412114 TI - Voluntary activation and cortical activity during a sustained maximal contraction: an fMRI study. AB - Motor fatigue is an exercise-induced reduction in the force-generating capacity. The underlying mechanisms can be separated into factors residing in the periphery or in the central nervous system. We designed an experiment in which we investigated central processes underlying motor fatigue by means of magnetic resonance imaging in combination with the twitch interpolation technique. Subjects performed a sustained maximal abduction (2 min) with the right index finger. Brain activation was recorded with an MR scanner, together with index finger abduction force, EMG of several hand muscles and interpolated twitches. Mean activity per volume was calculated for the primary motor cortex and the secondary motor areas (supplementary motor, premotor, and cingulate areas) as well as mean force and mean rectified EMG amplitude. Results showed a progressive decline in maximal index finger abduction force and EMG of the target muscles combined with an increase in brain activity in the contralateral primary motor cortex and secondary motor areas. Analysis of the twitches superimposed on the sustained contraction revealed that during the contraction the voluntary drive decreased significantly. In conclusion, our data showed that despite an increase in brain activity the voluntary activation decreased. This suggests that, although the CNS increased its input to the relevant motor areas, this increase was insufficient to overcome fatigue-related changes in the voluntary drive. PMID- 18412115 TI - Severe Marfan syndrome due to FBN1 exon deletions. AB - Marfan syndrome is an autosomal dominant condition, with manifestations mainly in the skeletal, ocular, and cardiovascular systems. The disorder is caused by mutations in fibrillin-1 gene (FBN1). The majority of these are family-specific point mutations, with a small number being predicted to cause exon-skipping. To date, there have only been five reports of in-frame exon deletions in FBN1, with the largest of these spanning three exons. Mosaicism is rarely recorded and has only been reported in the unaffected, or mildly affected, parents of probands. Here, we report on the clinical histories of two children with exon deletions in FBN1. Both have severe Marfan syndrome with significant signs in infancy. One patient has a deletion of exon 33, which has not previously been reported. The other has the largest reported deletion, which spans 37 exons, and also represents the first reported case of mosaicism in a patient with Marfan syndrome. PMID- 18412116 TI - Tracheomalacia in siblings with otopalatodigital syndrome. PMID- 18412117 TI - Mosaic FMR1 deletion causes fragile X syndrome and can lead to molecular misdiagnosis: a case report and review of the literature. AB - The most common cause of fragile X syndrome is expansion of a CGG trinucleotide repeat in the 5'UTR of FMR1. This expansion leads to transcriptional silencing of the gene. However, other mutational mechanisms, such as deletions of FMR1, also cause fragile X syndrome. The result is the same for both the expansion mediated silencing and deletion, absence of the gene product, FMRP. We report here on an 11-year-old boy with a cognitive and behavioral profile with features compatible with, but not specific to, fragile X syndrome. A mosaic deletion of 1,013,395 bp was found using high-density X chromosome microarray analysis followed by sequencing of the deletion breakpoints. We review the literature of FMR1 deletions and present this case in the context of other FMR1 deletions having mental retardation that may or may not have the classic fragile X phenotype. PMID- 18412118 TI - The clinical spectrum of homozygous HOXA1 mutations. AB - We describe nine previously unreported individuals from six families who have homozygous mutations of HOXA1 and either the Bosley-Salih-Alorainy syndrome (BSAS) or the Athabascan brainstem dysgenesis syndrome (ABDS). Congenital heart disease was present in four BSAS patients, two of whom had neither deafness nor horizontal gaze restriction, thus raising the possibility that cardiovascular malformations might be a clinically isolated, or relatively isolated, manifestation of homozygous HOXA1 mutations. Two ABDS probands had relatively mild mental retardation. These individuals blur the clinical distinctions between the BSAS and ABDS HOXA1 variants and broaden the phenotype and genotype of the homozygous HOXA1 mutation clinical spectrum. PMID- 18412119 TI - Delineation of a critical region on chromosome 18 for the del(18)(q12.2q21.1) syndrome. AB - Deletions involving the long arm of chromosome 18 have been reported in many patients. Most of these deletions are localized in the distal half of the long arm (18q21.1 --> qter) and are detectable by standard cytogenetic analysis. However, smaller interstitial deletions leading to a recognizable phenotype and residing in the region around chromosome band 18q12.3 (bands q12-q21) are less common. Here we report on an interstitial deletion of less than 1.8 Mb within chromosomal band 18q12.3. The phenotypic features of the propositus correspond well with those observed in patients with larger cytogenetically detectable deletions encompassing chromosome band 18q12.3. The deletion enabled us to define a critical region for the following features of the del(18)(q12.2q21.1) syndrome: hypotonia, expressive language delay, short stature, and behavioral problems. PMID- 18412120 TI - Report of a black South African child with oculodentodigital dysplasia and a novel GJA1 gene mutation. PMID- 18412121 TI - Cerebral impairment in chronic solvent-induced encephalopathy. AB - OBJECTIVE: Worldwide, many workers experience occupational exposure to organic solvents, which may induce chronic solvent-induced encephalopathy (CSE). Disturbances within the frontostriatothalamic (FST) circuitry might explain the symptomatology of CSE. We tested the hypothesis of FST circuitry abnormalities in CSE, as well as associations with performance of psychomotor speed, attention, and solvent exposure. To detect preclinical, solvent-related effects, we also studied the FST circuitry in solvent-exposed, but asymptomatic workers. METHODS: Ten CSE patients, 10 asymptomatic but solvent-exposed house painters (EC), and 11 nonexposed asymptomatic carpenters were included. Dopamine D(2) receptor (D2R) binding, central nervous system tissue metabolites, and fractional anisotropy were measured within the FST circuitry, using single-photon emission computed tomography, magnetic resonance spectroscopy, and diffusion tensor imaging. Performance of psychomotor speed and attention, and severity of solvent exposure were assessed. RESULTS: Striatal D2R binding was reduced in CSE. In the solvent exposed asymptomatic patients, striatal D2R binding and levels of N acetylaspartate + N-acetylaspartyl-glutamate in frontal gray matter were reduced. In both exposed groups, a trend was seen for reduced choline in frontal gray matter. In CSE, the fractional anisotropy in the thalamus, caudate nucleus, and striatal D2R binding significantly predicted reduced performance of attention and psychomotor speed. In CSE, striatal D2R binding showed a negative correlation with solvent exposure. INTERPRETATION: This is the first study in CSE showing pronounced disturbances within the FST circuitry that are related to the clinical findings and to exposure severity to solvents. The comparable, but milder, abnormalities within the FST circuitry in the exposed asymptomatic workers may imply a presymptomatic phase of CSE. PMID- 18412122 TI - Molecular aspects, clinical aspects and possible treatment modalities for Costello syndrome: Proceedings from the 1st International Costello Syndrome Research Symposium 2007. PMID- 18412123 TI - 12q12 deletion: a new patient contributing to genotype-phenotype correlation. PMID- 18412124 TI - Growth charts for patients affected with Morquio A disease. AB - Children with Morquio A disease grow poorly and become physically handicapped because of systemic bone disease. The purpose of this study was to describe observed growth patterns and their relationship with the physical condition of patients with Morquio A. In a one-center study, questionnaire-based longitudinal and cross sectional data were used to develop growth curves, to assess physical activity and to determine the incidence of surgical procedures in 354 patients with Morquio A. Mean birth lengths of boys and girls were 52.6 and 52.1 cm, respectively. The mean final heights for males and females at 18 years and older were 122.4 +/- 21.5 and 113.1 +/- 22.6 cm, respectively. These results corresponded to -7.4 SD for males and -7.7 SD for females compared to the normal healthy controls. Mean birth weights for boys and girls were 3.59 +/- 0.58 and 3.5 +/- 0.7 kg, respectively. The mean body mass index for males and females at over 18 years of age was 24.7 +/- 6.1 and 25.6 +/- 5.4 kg/m(2), respectively. The growth pattern in Morquio A patients was characterized by impaired growth velocity after 1 year of age. This is the first report providing growth charts for patients with Morquio A, which can help with monitoring the disease and assessing the clinical efficacy of treatments. PMID- 18412125 TI - Changes in germ cells population and in vitro oocyte maturation in female mice with imperforated vagina. AB - This study characterized female mice with a congenital defect in their reproductive tract. In females derived from an outbred colony maintained in the Department of Genetics and Evolution, the frequency of the imperforated vaginae was approximately 2%. A consequence of this defect is infertility. Affected animals developed hydrometrocolpos (the uterus and vagina were greatly distended by fluid). Morphology of the ovary and oviduct was normal and similar to that of control mice. In the females with an imperforated vagina numerous corpora lutea were observed. Accordingly, we have found that both spontaneous as well as exogenous gonadotrophin-induced ovulation occurs in such females. Nevertheless, oocytes derived from ovaries of occluded females exhibited a partial block during in vitro maturation. Histological analysis of ovarian tissue revealed an increase in the number of primary follicles (type I follicles) and a decrease in the number of secondary and antral follicles (type IV and type VI follicles) when compared with control mice. The concentration of androgens in the ovarian tissue was higher in the affected females. Our data show that females with an imperforated vagina can be a useful model for studying the mechanism of genetic control of the development of the urogenital tract in mammals. PMID- 18412126 TI - Atypical XX male with the SRY gene located at the long arm of chromosome 1 and a 1qter microdeletion. AB - Male individuals with a 46,XX karyotype have been designated as XX males. In 80% of the cases, the presence of Yp sequences, including the male sex-determining gene, SRY, has been demonstrated by molecular and/or fluorescence in situ hybridization (FISH) analyses. In most cases, Yp sequences are located on the short arm of the X chromosome, resulting from unequal recombination between Yp and Xp during paternal meiosis. Much less frequent in XX males is the localization of the SRY gene to an autosome. Here we report on the genetic investigation of an atypical XX male in which the SRY gene was located at the end of the long arm of chromosome 1. The patient, with a normal male phenotype, was referred for azoospermia. Conventional cytogenetic analysis showed a 46,XX karyotype. Molecular-cytogenetics (FISH) and molecular (PCR and MLPA) studies identified not only Yp-specific sequences located on the distal long arm of chromosome 1 but also the deletion of the subtelomeric 1qter region. A specific phenotype has been reported for a deletion of the 1qter region associated with mental retardation. The molecular investigation of the 1qter region showed that in our patient the microdeletion is more telomeric than in patients reported with mental retardation. To our knowledge, this is the first report of a XX male with the Yp region transferred to the terminal long arm of chromosome 1. This is also the first microdeletion of the subtelomeric 1qter region not associated with mental retardation. PMID- 18412127 TI - Conformational transitions of flanking purines in HIV-1 RNA dimerization initiation site kissing complexes studied by CHARMM explicit solvent molecular dynamics. AB - Dimerization of HIV-1 genomic RNA is initiated by kissing loop interactions at the Dimerization Initiation Site (DIS). Dynamics of purines that flank the 5' ends of the loop-loop helix in HIV-1 DIS kissing complex were explored using explicit solvent molecular dynamics (MD) simulations with the CHARMM force field. Multiple MD simulations (200 ns in total) of X-ray structures for HIV-1 DIS Subtypes A, B, and F revealed conformational variability of flanking purines. In particular, the flanking purines, which in the starting X-ray structures are bulged-out and stack in pairs, formed a consecutive stack of four bulged-out adenines at the beginning of several simulations. This conformation is seen in the crystal structure of DIS Subtype F with no interference from crystal packing, and was frequently reported in our preceding MD studies performed with the AMBER force field. However, as CHARMM simulations progressed, the four continuously stacked adenines showed conformational transitions from the bulged-out into the bulged-in geometries. Although such an arrangement has not been seen in any X-ray structure, it has been suggested by a recent NMR investigation. In CHARMM simulations, in the longer time scale, the flanking purines display the tendency to move to bulged-in conformations. This is in contrast with the AMBER simulations, which indicate a modest prevalence for bulged-out flanking base positions in line with the X-ray data. The simulations also suggest that the intermolecular stacking between purines from the opposite hairpins can additionally stabilize the kissing complex. PMID- 18412128 TI - Polymeric micelles anchored with TAT for delivery of antibiotics across the blood brain barrier. AB - Polymeric micelles self-assembled from cholesterol-conjugated poly(ethylene glycol) (PEG) and anchored with transcriptional activator TAT peptide (TAT-PEG-b Col) were fabricated for delivery of antibiotics across the blood-brain barrier (BBB). Ciprofloxacin, which demonstrated a high bactericidal effect, was efficiently loaded into the micelles by a membrane dialysis method. The ciprofloxacin-loaded micelles were characterized via dynamic light scattering and SEM. The micelles were spherical in nature, having an average diameter of smaller than 180 nm. Sustained release of ciprofloxacin was achieved over 6 h in phosphate-buffered saline (pH 7.4) at 37 degrees C. Confocal laser scanning microscopy reveals that the uptake of Fluorescein 5-isothiocyanate (FITC)-loaded TAT-PEG-b-Col micelles by human astrocytes was much higher than that of free FITC. Animal studies proved that these micelles crossed the BBB and entered the brain. The TAT-conjugated micelles may be used to deliver antibiotics across the BBB for treatment of brain infections. PMID- 18412129 TI - A new vibrator to stimulate muscle proprioceptors in fMRI. AB - Studying cognitive brain functions by functional magnetic resonance imaging (fMRI) requires appropriate stimulation devices that do not interfere with the magnetic fields. Since the emergence of fMRI in the 90s, a number of stimulation devices have been developed for the visual and auditory modalities. Only few devices, however, have been developed for the somesthesic modality. Here, we present a vibration device for studying somesthesia that is compatible with high magnetic field environments and that can be used in fMRI machines. This device consists of a poly vinyl chloride (PVC) vibrator containing a wind turbine and of a pneumatic apparatus that controls 1-6 vibrators simultaneously. Just like classical electromagnetic vibrators, our device stimulates muscle mechanoreceptors (muscle spindles) and generates reliable illusions of movement. We provide the fMRI compatibility data (phantom test), the calibration curve (vibration frequency as a function of air flow), as well as the results of a kinesthetic test (perceived speed of the illusory movement as a function of vibration frequency). This device was used successfully in several brain imaging studies using both fMRI and magnetoencephalography. PMID- 18412130 TI - Accessing newly learned names and meanings in the native language. AB - Ten healthy adults encountered pictures of unfamiliar archaic tools and successfully learned either their name, verbal definition of their usage, or both. Neural representation of the newly acquired information was probed with magnetoencephalography in an overt picture-naming task before and after learning, and in two categorization tasks after learning. Within 400 ms, activation proceeded from occipital through parietal to left temporal cortex, inferior frontal cortex (naming) and right temporal cortex (categorization). Comparison of naming of newly learned versus familiar pictures indicated that acquisition and maintenance of word forms are supported by the same neural network. Explicit access to newly learned phonology when such information was known strongly enhanced left temporal activation. By contrast, access to newly learned semantics had no comparable, direct neural effects. Both the behavioral learning pattern and neurophysiological results point to fundamentally different implementation of and access to phonological versus semantic features in processing pictured objects. PMID- 18412131 TI - Nonrigid registration of multiple sclerosis brain images using lesion inpainting for morphometry or lesion mapping. AB - Morphometric studies of medical images often include a nonrigid registration step from a subject to a common reference. The presence of white matter multiple sclerosis lesions will distort and bias the output of the registration. In this article, we present a method to remove this bias by filling such lesions to make the brain look like a healthy brain before the registration. We finally propose a dedicated method to fill the lesions and present numerical results showing that our method outperforms current state of the art method. PMID- 18412132 TI - White matter tract integrity in aging and Alzheimer's disease. AB - The pattern of degenerative changes in the brain white matter (WM) in aging, mild cognitive impairment (MCI), and Alzheimer's disease (AD) has been under debate. Methods of image analysis are an important factor affecting the outcomes of various studies. Here we used diffusion tensor imaging (DTI) to obtain fractional anisotropy (FA) measures of the WM in healthy young (n = 8), healthy elderly (n = 22), MCI (n = 8), and AD patients (n = 16). We then applied "tract-based spatial statistics" (TBSS) to study the effects of aging, MCI, and AD on WM integrity. Our results show that changes in WM integrity (that is, decreases in FA) are different between healthy aging and AD: in healthy older subjects compared with healthy young subjects decreased FA was primarily observed in frontal, parietal, and subcortical areas whereas in AD, compared with healthy older subjects, decreased FA was only observed in the left anterior temporal lobe. This different pattern of decreased anatomical connectivity in normal aging and AD suggests that AD is not merely accelerated aging. PMID- 18412133 TI - A cross-modal system linking primary auditory and visual cortices: evidence from intrinsic fMRI connectivity analysis. AB - Recent anatomical and electrophysiological evidence in primates indicates the presence of direct connections between primary auditory and primary visual cortex that constitute cross-modal systems. We examined the intrinsic functional connectivity (fcMRI) of putative primary auditory cortex in 32 young adults during resting state scanning. We found that the medial Heschl's gyrus was strongly coupled, in particular, to visual cortex along the anterior banks of the calcarine fissure. This observation was confirmed using novel group-level, tensor based independent components analysis. fcMRI analysis revealed that although overall coupling between the auditory and visual cortex was significantly reduced when subjects performed a visual perception task, coupling between the anterior calcarine cortex and auditory cortex was not disrupted. These results suggest that primary auditory cortex has a functionally distinct relationship with the anterior visual cortex, which is known to represent the peripheral visual field. Our study provides novel, fcMRI-based, support for a neural system involving low level auditory and visual cortices. PMID- 18412134 TI - Giving speech a hand: gesture modulates activity in auditory cortex during speech perception. AB - Viewing hand gestures during face-to-face communication affects speech perception and comprehension. Despite the visible role played by gesture in social interactions, relatively little is known about how the brain integrates hand gestures with co-occurring speech. Here we used functional magnetic resonance imaging (fMRI) and an ecologically valid paradigm to investigate how beat gesture a fundamental type of hand gesture that marks speech prosody-might impact speech perception at the neural level. Subjects underwent fMRI while listening to spontaneously-produced speech accompanied by beat gesture, nonsense hand movement, or a still body; as additional control conditions, subjects also viewed beat gesture, nonsense hand movement, or a still body all presented without speech. Validating behavioral evidence that gesture affects speech perception, bilateral nonprimary auditory cortex showed greater activity when speech was accompanied by beat gesture than when speech was presented alone. Further, the left superior temporal gyrus/sulcus showed stronger activity when speech was accompanied by beat gesture than when speech was accompanied by nonsense hand movement. Finally, the right planum temporale was identified as a putative multisensory integration site for beat gesture and speech (i.e., here activity in response to speech accompanied by beat gesture was greater than the summed responses to speech alone and beat gesture alone), indicating that this area may be pivotally involved in synthesizing the rhythmic aspects of both speech and gesture. Taken together, these findings suggest a common neural substrate for processing speech and gesture, likely reflecting their joint communicative role in social interactions. PMID- 18412135 TI - "Severe" wear challenge to 36 mm mechanically enhanced highly crosslinked polyethylene hip liners. AB - Our purpose was to compare the wear performance of mechanically enhanced 5Mrad highly crosslinked polyethylene (MEP, ArComXL) hip liners to (control) 3Mrad UHMWPE liners (ArCom) in 36 mm head size. As a more severe synergy of clinically relevant test models, we contrasted wear with custom roughened Co-Cr surfaces (Ra 500 nm) to the standard pristine Co-Cr heads (Ra < 20 nm) using a severe microseparation test mode in our hip simulator. We adopted a previously published model to estimate potential biological activity. On new Co-Cr heads, the MEP liners showed a 47% reduction in volumetric wear a 13% reduction in wear particle size and a 27% reduction in Functional Biological Activity (FBA) compared to our control. On rough Co-Cr heads, the MEP liners showed little advantage in terms of volumetric wear compared with the control. However, the MEP liners overall showed a 38% reduction in FBA compared to the control owing to a larger volume fraction of larger particles. Thus overall the MEP liners appeared to offer advantages in terms of reduced FBA indices. PMID- 18412136 TI - Instability of self-assembled monolayers as a model material system for macrophage/FBGC cellular behavior. AB - Novel self-assembled monolayers (SAMs) designed to present homogenous surface chemistries were utilized to further investigate the material surface chemistry dependent macrophage and foreign-body giant cell (FBGC) behaviors, including macrophage adhesion, fusion, and apoptosis. Contact angle analysis revealed instabilities in the --CH(3) and --COOH terminated SAM surfaces upon incubation in serum-free media (SFM) at 37 degrees C or under dry, room temperature conditions. Further analysis indicated that the --CH(3) terminated SAM surface degraded rapidly within 2 h and loss of sufficient SAM units to be comparable to the gold (Au) control surface, within 24 h of incubation in SFM at 37 degrees C. After 5 days of incubation in SFM at 37 degrees C, the contact angles for the - COOH terminated SAM surfaces increased markedly. AFM analysis confirmed the desorption of --CH(3) terminated SAM molecules from the surface with increased roughness and marked appearance of peaks and valleys within 2 h. A decrease in the thickness of the --COOH terminated SAM surface also suggests molecular desorption over time. No significant changes in contact angle or AFM analyses were observed on the --OH terminated SAM surfaces. Cellular adhesion decreased more rapidly on the Au control and --CH(3) terminated SAM surfaces in comparison to the other surfaces. However by day 10, cellular adhesion, fusion, and apoptosis were comparable on all SAM surfaces and the Au control. These studies suggest that SAM surfaces may not be suitable for long-term studies where material dependent properties are investigated. PMID- 18412137 TI - Co-expression of elastin and collagen leads to highly compliant engineered blood vessels. AB - Elastin synthesis and physiologic compliance are significant challenges in blood vessel tissue engineering. Here, we report that a biocompatible elastomeric scaffold can support the co-expression of elastin and collagen, which likely yielded the physiologic compliance in the constructs. A biodegradable elastomer, poly(glycerol sebacate), was fabricated into highly porous tubular scaffolds. Primary baboon arterial smooth muscle cells (SMCs) were seeded in the lumen of the scaffolds followed by a 1-week culture under gentle perfusion. Circulating endothelial progenitor cells (EPCs) isolated from baboon peripheral blood was seeded directly on the smooth muscle layer in the lumen on day 8. The constructs were perfused using a pulsatile flow system for another 2 weeks before characterization. In another set of experiments, the SMCs were cultured for 7 weeks and were co-cultured for 1 week with the EPCs. Constructs obtained using either set of culture conditions contained elastin and collagen: Masson's trichrome stain showed a circumferential collagen band in the constructs, and elastin was evident from its characteristic autofluorescence, Verhoff's stain, and amino acid analysis of insoluble remnants after hot alkali digestion. All constructs had a confluent cellular lumen with cells well-dispersed throughout the scaffolds. At physiologic pressures, the compliance of the 8-week construct was comparable to human arteries as observed in pressure-diameter testing. Combination of elastomeric scaffolds, co-culture of EPC and SMC, and mechanical conditioning appears to encourage the expression of a more natural extracellular matrix and lead to physiologically-relevant compliance; both are major challenges in blood vessel tissue engineering. PMID- 18412138 TI - Effect of short-range forces on the length distribution of fibrous cytoskeletal proteins. AB - The length distribution of cytoskeletal filaments is an important physical parameter, which can modulate physiological cell functions. In both eukaryotic and prokaryotic cells various biological cytoskeletal polymers form supramolecular structures due to short-range forces induced mainly by molecular crowding or cross linking proteins, but their in vivo length distribution remains difficult to measure. In general, based on experimental evidence and mathematical modeling of actin filaments in aqueous solutions, the steady state length distribution of fibrous proteins is believed to be exponential. We performed in vitro TIRF- and electron-microscopy to demonstrate that in the presence of short range forces, which are an integral part of any living cell, the steady state length distributions of the eukaryotic cytoskeletal biopolymer actin, its prokaryotic homolog ParM and microtubule homolog FtsZ deviate from the classical exponential and are either double-exponential or Gaussian, as recent theoretical modeling predicts. Double exponential or Gaussian distributions opposed to exponential can change for example the visco-elastic properties of actin networks within the cell, influence cell motility by decreasing the amount of free ends at the leading edge of the cell or effect the assembly of FtsZ into the bacterial Z ring thus modulating membrane constriction. PMID- 18412139 TI - Multiscale modeling of nucleic acids: insights into DNA flexibility. AB - The elastic rod theory is used together with all-atom normal mode analysis in implicit solvent to characterize the mechanical flexibility of duplex DNA. The bending, twisting, stretching rigidities extracted from all-atom simulations (on linear duplexes from 60 to 150 base pairs in length and from 94-bp minicircles) are in reasonable agreement with experimental results. We focus on salt concentration and sequence effects on the overall flexibility. Bending persistence lengths are about 20% higher than most experimental estimates, but the transition from low-salt to high-salt behavior is reproduced well, as is the dependence of the stretching modulus on salt (which is opposite to that of bending). CTG and CGG trinucleotide repeats, responsible for several degenerative disorders, are found to be more flexible than random DNA, in agreement with several recent studies, whereas poly(dA).poly(dT) is the stiffest sequence we have encountered. The results suggest that current all-atom potentials, which were parameterized on small molecules and short oligonucleotides, also provide a useful description of duplex DNA at much longer length scales. PMID- 18412140 TI - Methods of evaluating human exposure to electromagnetic fields radiated from operating base stations in Korea. AB - This article presents measurement methods used to determine the human exposure to electromagnetic fields radiated from operating base stations. In Korea, when evaluating the human exposure to electromagnetic fields from operating base stations, the measurement procedure is different between the following cases: in situ measurement and electromagnetic environment measurement. When performing an in situ measurement, compliance with human exposure limits is determined by the spatially averaged field value obtained within the space occupied by humans at one arbitrary position, but when performing an electromagnetic environment measurement, it is determined by the maximum value at the highest field position selected from several places. PMID- 18412141 TI - Epstein-Barr virus latent membrane protein 1 mediates serine 25 phosphorylation and nuclear entry of annexin A2 via PI-PLC-PKCalpha/PKCbeta pathway. AB - We have previously elucidated that Epstein-Barr-virus-encoded latent membrane protein 1 (LMP1) can increase the serine phosphorylation level of annexin A2 by activating the protein kinase C (PKC) signaling pathway and that LMP1 induces the nuclear entry of annexin A2 in an energy- and temperature-dependent manner. Here, we further confirm that LMP1 increases the serine phosphorylation level of annexin A2 by activating the phosphoinositide-specific phospholipase C (PI-PLC) PKC alpha/PKC beta pathway, mainly through the activation of the PKCbeta pathway. Additionally, active recombinant PKC alpha, PKC beta I, and PKC beta II kinases are able to phosphorylate annexin A2 in vitro. Annexin A2 in the nucleus plays an important role in DNA synthesis and cell proliferation. By site-specific substitution of glutamic acid in the place of serine 11 and 25 in the N-terminus, we show that serine 25 phosphorylation of annexin A2 was associated with the nuclear entry of annexin A2, DNA synthesis and cell proliferation, whereas serine 11 has no obvious influence. We demonstrate for the first time that the PI-PLC PKCalpha/PKCbeta pathway plays an important role in serine phosphorylation and in the nuclear entry of annexin A2 mediated by LMP1. In addition, we show that annexin A2 is the substrate protein of PKC alpha, PKC betaI, and PKC betaII kinases. Serine 25 phosphorylation of annexin A2 is shown to be associated with its nuclear entry, DNA synthesis, and cell proliferation. PMID- 18412142 TI - Fibroblast growth factor-2 regulates expression of osteopontin in periodontal ligament cells. AB - Osteopontin is a protein found in the bone-related matrix and plays multiple regulatory roles in mineralizing and non-mineralizing tissue. In osteogenic cell lines, the expression of osteopontin increases with the progression of differentiation, but both the expression and function of osteopontin vary with the cell type and its activation state. In this study, we examined the expression of osteopontin by clones established from mouse periodontal ligament, in response to inorganic phosphate and fibroblast growth factor (FGF)-2, which can induce periodontal tissue regeneration. The involvement of inorganic phosphate in the expression of osteopontin during the course of cell differentiation of a clone MPDL22 was confirmed by addition of foscarnet, an inorganic phosphate transport inhibitor. Although FGF-2 decreased the mRNA expression of almost every bone related protein in MPDL22, FGF-2 upregulated the expression of osteopontin in MPDL22 at both mRNA and protein levels. Interestingly, FGF-2 enhanced the concentration of osteopontin in the culture supernatant of MPDL22, whereas inorganic phosphate did not. The FGF-2-induced osteopontin in the culture supernatant seems to be involved in cell survival activity. An immunohistochemical study showed that the FGF-2-induced osteopontin was mainly present in perinuclear matrices while the inorganic phosphate-induced osteopontin was associated with extracellular matrices in addition to perinuclear matrices. The present results indicated that FGF-2 induces unique expression of osteopontin, which may play a role different from the other bone-related proteins during the process of periodontal tissue regeneration by FGF-2. PMID- 18412143 TI - The role of p38 MAPK and JNK in Arsenic trioxide-induced mitochondrial cell death in human cervical cancer cells. AB - Previously, we have shown that the release of AIF from mitochondria is required for As2O3-induced cell death in human cervical cancer cells, and that reactive oxygen species (ROS) is necessary for AIF release from mitochondria. In this study, we further investigated the role of MAPKs in ROS-mediated mitochondrial apoptotic cell death triggered by As2O3. As2O3-induced apoptotic cell death in HeLa cells was associated with activation and mitochondrial translocation of Bax, a marked phosphorylation of Bcl-2, reduction of Bcl-2 and Bax interaction, dissipation of mitochondrial membrane potential. Using small interfering RNA, reduced Bax expression effectively attenuated As2O3-induced mitochondrial membrane potential loss and apoptotic cell death. Moreover, the phosphorylation of Bcl-2 induced by As2O3 diminished its ability to bind to Bax. Treatment of cells with As2O3 activated both the p38 MAPK and JNK pathways. Mitochondrial translocation of Bax was completely suppressed in the presence of p38 MAPK inhibitor PD169316 or si-p38 MAPK. The As2O3-induced Bcl-2 phosphorylation was attenuated largely by JNK inhibition using SP600125 or si-JNK and to some extent by p38 MAPK inhibition with PD169316 or si-p38 MAPK. In addition, N-acetyl-L cystein (NAC), a thiol-containing anti-oxidant, completely blocked As2O3-induced p38 MAPK and JNK activations, mitochondria translocation of Bax, and phosphorylation of Bcl-2. These results support a notion that ROS-mediated activations of p38 MAPK and JNK in response to As2O3 treatment signals activation of Bax and phosphorylation of Bcl-2, resulting in mitochondrial apoptotic cell death in human cervical cancer cells. PMID- 18412144 TI - Quantification of myocardial iron overload by cardiovascular magnetic resonance imaging T2* and review of the literature. AB - Heart failure due to myocardial iron overload remains the leading cause of death in patients with transfusion-dependent anemias. Iron overload-induced cardiomyopathy is reversible if intensive chelation therapy is instituted on time. Thus, early detection of myocardial iron deposition is imperative to prevent overt heart failure. Conventional cardiac monitoring, including physical examination, electrocardiography, echocardiography or serum ferritin levels fail to predict manifest or subclinical myocardial involvement resulting from iron overload. Cardiovascular magnetic resonance imaging T2* (cMRI-T2*, pronounced T2 star) times correlate well with myocardial iron levels. This timely review focuses on the utility of cMRI-T2*, for the preclinical detection of myocardial iron overload and monitoring of myocardial iron content during chelation therapy. PMID- 18412145 TI - Rapid normalization of a highly thickened pericardium by chemotherapy in a patient with T-cell acute lymphoblastic lymphoma. AB - The most common tumor that affects the pericardium is malign lymphoma. T-cell lymphoblastic lymphoma (TLL) is a rare type of malign lymphomas. In this manuscript, we are reporting a patient with TLL with pericardial involvement diagnosed incidentally during the evaluation of pleural effusion. Echocardiographic examination showed thickened pericardium and pericardial effusion. The pericardial thickness was found to be 13 mm by computerized tomography and confirmed by echocardiography. The patient had systemic chemotherapy for TLL. On day 30 of chemotherapy, computerized tomography of the thorax and echocardiographic examination revealed normal pericardial thickness and minimal pericardial effusion. PMID- 18412146 TI - Antiperoxidative, antithyroidal, antihyperglycemic and cardioprotective role of Citrus sinensis peel extract in male mice. AB - An extract of Citrus sinensis (CS) peel was evaluated for its efficacy in ameliorating L-thyroxine (L-T(4)) induced tissue lipid peroxidation (LPO), hyperthyroidism and hyperglycemia in mice. In a preliminary investigation, of the three different doses of CS (12.5 mg/kg, 25 mg/kg and 50.0 mg/kg) peel extract, 25 mg/kg was found to be the most effective and antiperoxidative, while 50 mg/kg was proved to be hepatotoxic. Therefore in the pilot experiment the effects of 25 mg/kg/day of CS for 10 days were studied in L-T(4) induced hyperthyroid animals. L-T(4) (500 microg/kg/day for 10 days) increased the levels of thyroxine (T(4)) and triiodothyronine (T(3)) with a concomitant increase in serum glucose concentration, alpha-amylase activity, heart/body weight ratio (HW/BW), kidney/body weight ratio (KW/BW) and cardiac as well as hepatic LPO. However, it decreased the concentration of different serum lipids such as total cholesterol (TC), triglycerides (TG), high-density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C) and very low-density lipoprotein cholesterol (VLDL-C). Administration of CS extract (25 mg/kg/day) in hyperthyroid animals reversed most of these observations revealing the ameliorating potential of CS extract against various adverse effects of hyperthyroidism. It appears that the test extract primarily acts through its antioxidative/free radical scavenging, antithyroidal and HDL-C stimulating properties. PMID- 18412147 TI - Antiinflammatory effect of Daesiho, a Korean traditional prescription for cerebral infarct patients. AB - Daesiho, a prescription composed of eight herbal mixtures, has been widely used in the treatment of cerebral infarct in Oriental medicine. However, the mechanisms by which the formula affects the production of pro-inflammatory cytokines in cerebral infarct patients remains unknown. The levels of secretory protein pro-inflammatory cytokines, including tumor necrosis factor (TNF)-alpha, interleukin (IL)-1beta and IL-6, were significantly increased in both lipopolysaccharide (LPS) and phytohemagglutinin (PHA)-stimulated peripheral blood mononuclear cells (PBMCs) from cerebral infarct patients and LPS-stimulated THP-1 differentiated macrophage-like cells (THP-1/M). However, pretreatment with Daesiho significantly inhibited the secretion of pro-inflammatory cytokines, including TNF-alpha, IL-1beta, and IL-6, in stimulated PBMCs and THP-1/M cells. In addition, Daesiho significantly suppressed mRNA expression of pro-inflammatory cytokines. Therefore, these data indicate that Daesiho may be beneficial in the cessation of inflammatory processes of cerebral infarction through suppression of the production of pro-inflammatory cytokines via inhibition of mRNA expression. PMID- 18412148 TI - Effects of Chlorophytum borivilianum on sexual behaviour and sperm count in male rats. AB - This study was designed to evaluate the aphrodisiac and spermatogenic potential of the aqueous extract of dried roots of Chlorophytum borivilianum (CB) in rats. Male Wistar albino rats were divided into four groups. Rats were orally treated with (1) CONTROL GROUP: distilled water; (2) CB 125 mg/kg/day; (3) CB 250 mg/kg/day; and (4) Viagra((R)) group: 4 mg/kg/day sildenafil citrate and their sexual behaviour was monitored 3 h later using a receptive female. Their sexual behaviour was evaluated on days 1, 7, 14, 21 and 28 of treatment by pairing with a pro-oestrous female rat. For sperm count the treatment was continued further in all groups except the Viagra((R)) group for 60 days. At 125 mg/kg, CB had a marked aphrodisiac action, increased libido, sexual vigor and sexual arousal. Similarly, at the higher dose (250 mg/kg) all the parameters of sexual behaviour were enhanced, but showed a saturation effect after day 14. On day 60 the sperm count increased significantly in both the CB groups, 125 mg/kg and 250 mg/kg, in a dose dependent manner. Thus, roots of Chlorophytum borivilianum can be useful in the treatment of certain forms of sexual inadequacies, such as premature ejaculation and oligospermia. PMID- 18412149 TI - Gemcitabine plus oxaliplatin (GEMOX) combined with cetuximab in patients with progressive advanced stage hepatocellular carcinoma: results of a multicenter phase 2 study. AB - BACKGROUND: The authors conducted a phase 2 trial of the antiepidermal growth factor receptor (EGFR) monoclonal antibody cetuximab in combination with the gemcitabine plus oxaliplatin (GEMOX) regimen in patients with documented progressive hepatocellular carcinoma (HCC). METHODS: Forty-five untreated patients with advanced-stage progressive HCC were prospectively enrolled. Treatment consisted of cetuximab at a dose of 400 mg/m2 initially then 250 mg/m2 weekly, plus gemcitabine at a dose of 1000 mg/m2 on Day 1 and oxaliplatin at a dose of 100 mg/m2 on Day 2, every 2 weeks. Treatment was continued until disease progression, unacceptable toxicity, or patient refusal. RESULTS: Overall, 306 cycles were administered. Grade 3 to 4 hematologic toxicity consisted of thrombocytopenia (24%), neutropenia (20%), and anemia (4%). Grade 3 oxaliplatin induced neurotoxicity occurred in 5 patients (11%) and grade 3 cutaneous toxicity in 7 patients (16%). There were no treatment-related deaths. The confirmed response rate was 20% and disease stabilization was obtained in 40% of patients. The median progression-free and overall survival times were 4.7 months and 9.5 months, respectively. The 1-year survival rate was 40%. CONCLUSIONS: In poor prognosis patients with progressive advanced-stage HCC, the GEMOX-cetuximab combination appears to be active and to have manageable toxicity. A comparative randomized trial is now being planned. PMID- 18412150 TI - Measuring the significance of workload on performance of cytotechnologists in gynecologic cytology: a study using rapid prescreening. AB - BACKGROUND: Workload is extensively regulated and often used as a measure of quality in gynecologic cytology. Whether workload correlates with the sensitivity of screening in gynecologic cytology is not known. METHODS: The sensitivity of gynecologic cytology screening was measured over an 8-month period using the result of full screening coupled with the results of rapid prescreening. Sensitivity results were then correlated with daily workload volumes and the experience level of individual cytotechnologists. RESULTS: Rapid prescreening had an average sensitivity of 41.9% for atypical squamous cells of undetermined significance (ASCUS) and above. Full screening had a corrected sensitivity of 82.2% for ASCUS and above. Rapid prescreening increased the sensitivity of the laboratory to 89.9%. The sensitivity of full screening was significantly different between cytotechnologists (79.2% vs 99%, P < .001), but was not correlated with years of experience, sensitivity of rapid prescreening, or workload (all P > .05). When sensitivity and workload were examined on a monthly basis, there was no significant difference between sensitivity either as a group or individually at the highest and lowest workloads (P > .40 for all). CONCLUSIONS: Screeners sensitivity in gynecologic cytology appears to be unrelated to the experience level of individual cytotechnologists or to their workload at the levels examined. PMID- 18412151 TI - Activity against Mycobacterium smegmatis and M. tuberculosis by extract of South African medicinal plants. AB - Seven ethnobotanically selected medicinal plants were screened for their antimycobacterial activity. The minimum inhibitory concentration (MIC) of four plants namely Artemisia afra, Dodonea angustifolia, Drosera capensis and Galenia africana ranged from 0.781 to 6.25 mg/mL against Mycobacterium smegmatis. G. africana showed the best activity exhibiting an MIC of 0.78 mg/mL and a minimum bactericidal concentration (MBC) of 1.56 mg/mL. The MICs of ethanol extracts of D. angustifolia and G. africana against M. tuberculosis were found to be 5.0 and 1.2 mg/mL respectively. The mammalian cytotoxicity IC(50) value of the most active antimycobacterial extract, from G. africana, was found to be 101.3 microg/mL against monkey kidney Vero cells. Since the ethanol G. africana displayed the best antimycobacterial activity, it was subjected to fractionation which led to the isolation of a flavone, 5,7,2'-trihydroxyflavone. The MIC of this compound was found to be 0.031 mg/mL against M. smegmatis and 0.10 mg/mL against M. tuberculosis. This study gives some scientific basis to the traditional use of these plants for TB-related symptoms. PMID- 18412152 TI - The evolution of resistant Candida species in cancer centers: implications for treatment and prophylaxis. PMID- 18412153 TI - The changing epidemiology of invasive candidiasis: Candida glabrata and Candida krusei as the leading causes of candidemia in hematologic malignancy. AB - BACKGROUND: The objective of the current retrospective study was to compare the epidemiology of candidemia and its risk factors in patients who had hematologic malignancies(HM) with those in patients who had solid tumors (ST). METHODS: The medical and electronic records of all patients with cancer who had candidemia at the authors' institution from 1993 to 2003 were reviewed for demographic data and clinical information, including the use of prophylactic fluconazole, the infecting Candida species, and the source of candidemia (catheter-related vs other apparent sources). RESULTS: Six hundred thirty-five patients with candidemia were analyzed. C. glabrata and C. krusei were the leading causes of candidemia in 31% and 24% of patients with HM, respectively, and in 18% and 2% of patients with ST, respectively (P < .001). A catheter was the source of candidemia in 36% of the patients with ST and in 12% of the patients with HM (P < .001). Response to antifungal therapy occurred in 73% of the ST group compared with 49% of the HM group (P < .001). Multivariate logistic regression analysis revealed that fluconazole prophylaxis was a risk factor for both C. glabrata and C. krusei candidemia. The analysis also identified neutropenia as a risk factor for all candidemia and catheter-related infection as a risk factor for C. parapsilosis candidemia. CONCLUSIONS: The results of this study indicated that C. glabrata and C. krusei were the leading causes of candidemia in patients with HM. Neutropenia was the leading risk factor for all candidemia, whereas the catheter was the leading risk factor for C. parapsilosis candidemia. PMID- 18412154 TI - Oncofetal protein IMP3: a novel molecular marker that predicts metastasis of papillary and chromophobe renal cell carcinomas. AB - BACKGROUND: Whether an oncofetal protein, IMP3, can serve as a prognostic biomarker to predict metastasis for patients with localized papillary and chromophobe subtypes of renal cell carcinomas (RCCs) was investigated. METHODS: The expression of IMP3 in 334 patients with primary papillary and chromophobe RCC from multiple medical centers was evaluated by immunohistochemistry. The 317 patients with localized papillary and chromophobe RCCs were further evaluated for outcome analyses. RESULTS: IMP3 was significantly increased in a subset of localized papillary and chromophobe RCCs that subsequently metastasized. Patients with localized IMP3-positive tumors (n=33; 10%) were over 10 times more likely to metastasize (risk ratio [RR], 11.38; 95% confidence interval [CI], 5.40-23.96; P<.001) and were nearly twice as likely to die (RR, 1.91; 95% CI, 1.13-3.22; P=.016) compared with patients with localized IMP3 negative tumors. The 5-year metastasis-free and overall survival rates were 64% and 58% for patients with IMP3-positive localized papillary and chromophobe RCCs compared with 98% and 85% for patients with IMP3 negative tumors, respectively. In multivariable analysis adjusting for the TNM stage and nuclear grade, patients with IMP3-positive tumors were still over 10 times more likely to progress to distant metastasis (RR, 13.45; 95% CI, 6.00-30.14; P<.001) and were still nearly twice as likely die (RR, 1.95; 95% CI, 1.15-3.31; P=.013) compared with patients with IMP3-negative tumors. CONCLUSIONS: IMP3 is an independent prognostic biomarker that can be used to identify a subgroup of patients with localized papillary and chromophobe RCC who are at high risk for developing distant metastasis. PMID- 18412155 TI - Two pathologic types of hepatocellular carcinoma with lymph node metastasis with distinct prognosis on the basis of CK19 expression in tumor. AB - BACKGROUND: Few studies have investigated the pathologic types and prognosis of hepatocellular carcinoma (HCC) with lymph node metastasis (LNM). The purpose was to explore pathologic types and pertinent therapy of HCC with LNM. METHODS: An immunohistochemical study for CK19 and OV-6 was performed on tissue microarrays of HCC with LNM (n=47) and those without LNM (n=125). The clinicopathologic factors and patient survival were analyzed. RESULTS: Immunopositivity of CK19 and OV-6 in HCC with LNM were higher than that in 125 HCC without LNM (27.7% vs 5.6%, P=.000; 29.8% vs 12.8%, P=.009); their expressions were significantly correlated in HCC with LNM (correlation coefficient: 0.637, P=.000). The CK19 expression and tumor (T) classification of American Joint Committee on Cancer (AJCC)/International Union Against Cancer (UICC) tumor staging system were 2 independent risk factors for developing LNM (odds ratio [OR], 5.170, 95% confidence interval [CI], 1.840-14.528, P=.002; OR, 1.879, 95% CI, 1.236-2.857, P=.003). The CK19(+) group had shorter median survival (7.7 months vs 21.7 months, P=.013); CK19 expression was the independent prognostic factor for overall survival in HCC with LNM and was correlated with proliferating cell nuclear antigen labeling index and matrix metalloproteinase-9 expression (correlation coefficient: 0.484, P=.001 and 0.459, P=.001, respectively). CONCLUSIONS: CK19 expression and AJCC/UICC T classification were 2 independent risk factors for developing LNM in HCC. CK19 expression was the independent prognostic factor for HCC with LNM. It is of clinical significance for treatment modalities to differentiate HCC with intrahepatic cholangiocarcinoma-like differentiation (CK19[+]) from one with a higher T classification (CK19[-]). PMID- 18412156 TI - Dynamic compartmentalization of protein tyrosine phosphatase receptor Q at the proximal end of stereocilia: implication of myosin VI-based transport. AB - Hair cell stereocilia are apical membrane protrusions filled with uniformly polarized actin filament bundles. Protein tyrosine phosphatase receptor Q (PTPRQ), a membrane protein with extracellular fibronectin repeats has been shown to localize at the stereocilia base and the apical hair cell surface, and to be essential for stereocilia integrity. We analyzed the distribution of PTPRQ and a possible mechanism for its compartmentalization. Using immunofluorescence we demonstrate that PTPRQ is compartmentalized at the stereocilia base with a decaying gradient from base to apex. This distribution can be explained by a model of transport directed toward the stereocilia base, which counteracts diffusion of the molecules. By mathematical analysis, we show that this counter transport is consistent with the minus end-directed movement of myosin VI along the stereocilia actin filaments. Myosin VI is localized at the stereocilia base, and exogenously expressed myosin VI and PTPRQ colocalize in the perinuclear endosomes in COS-7 cells. In myosin VI-deficient mice, PTPRQ is distributed along the entire stereocilia. PTPRQ-deficient mice show a pattern of stereocilia disruption that is similar to that reported in myosin VI-deficient mice, where the predominant features are loss of tapered base, and fusion of adjacent stereocilia. Thin section and freeze-etching electron microscopy showed that localization of PTPRQ coincides with the presence of a dense cell surface coat. Our results suggest that PTPRQ and myosin VI form a complex that dynamically maintains the organization of the cell surface coat at the stereocilia base and helps maintain the structure of the overall stereocilia bundle. PMID- 18412157 TI - Continuous generation of colitogenic CD4(+) T cells in persistent colitis. AB - Inflammatory bowel diseases take chronic courses due to the expansion of colitogenic CD4(+) cells. However, it is unclear whether the persistent disease is driven by continuous reactivation of colitogenic memory CD4(+) cells to generate effector CD4(+) cells or by continuous generation of effector CD4(+) cells from naive cells. To clarify this issue, we performed a series of sequential adoptive transfers of Ly5.2(+) and Ly5.1(+) CD4(+)CD45RB(high) cells into RAG-2(-/-) mice at different time points. We show here that the secondarily transferred CD4(+)CD45RB(high) cells can be converted to CD4(+)CD44(high)CD62L( )IL-7Ralpha(high) effector-memory T cells even in the presence of pre-existing effector-memory CD4(+) cells. Although the total cell numbers of CD4(+) cells in established colitic mice were consistently equivalent irrespective of the number of primarily transferred cells, the ratio of primarily and secondarily transferred cells was dependent on the ratio of the transferred cell numbers, but not on the order of the transfer. Of note, we found that primarily transferred CD4(+) cells produced significantly lower amounts of IFN-gamma and IL-17 than CD4(+) cells arising from secondary transfer. In conclusion, the continuous generation of colitogenic CD4(+) cells that compensate for exhausted CD4(+) cells may be one of the mechanisms involved in the persistence of colitis. PMID- 18412158 TI - Interferon-gamma limits Th1 lymphocyte adhesion to inflamed endothelium: a nitric oxide regulatory feedback mechanism. AB - CD4(+) T helper (Th1 and Th2) cell localization to a site of inflammation is important for the development, maintenance and regulation of an immune response. The factors that regulate Th1 and Th2 cell recruitment into tissue are not fully understood. The aim of the present study was to examine the effect of different cytokine microenvironments on the recruitment of Th1 and Th2 lymphocytes into tissue. Fluorescently labelled Th1 or Th2 lymphocyte-endothelial interactions were observed via intravital microscopy of the cytokine-treated cremaster muscle. Our results show that TNF-alpha alone is sufficient to maximally recruit Th1 cells. Surprisingly, treatment with TNF-alpha + IFN-gamma significantly decreased Th1 adhesion and emigration in comparison to TNF-alpha treatment alone. The decreased adhesion of Th1 cells in response to TNF-alpha + IFN-gamma reflected a decreased ability to bind to ICAM-1 and was iNOS-dependent. This phenomenon was not observed with Th2 cells. These results suggest that IFN-gamma may play a key immunomodulatory role in the recruitment of different T lymphocyte subsets. Indeed, blockade of IFN-gamma or iNOS function during the Th1-mediated contact hypersensitivity response resulted in an acceleration and exacerbation of the late-phase inflammatory response. PMID- 18412159 TI - Endoplasmic reticulum stress and the unfolded protein response are linked to synergistic IFN-beta induction via X-box binding protein 1. AB - Type I IFN are strongly induced upon engagement of certain pattern recognition receptors by microbial products, and play key roles in regulating innate and adaptive immunity. It has become apparent that the endoplasmic reticulum (ER) stress-induced unfolded protein response (UPR), in addition to restoring ER homeostasis, also influences the expression of certain inflammatory cytokines. However, the extent to which UPR signaling regulates type I IFN remains unclear. Here we show that cells undergoing a UPR respond to TLR4 and TLR3 ligands, and intracellular dsRNA, with log-fold greater IFN-beta induction. This synergy is not dependent on autocrine type I IFN signaling, but unexpectedly requires the UPR transcription factor X-box binding protein 1 (XBP-1). Synergistic IFN-beta induction also occurs in HLA-B27/human beta(2)m-transgenic rat macrophages exhibiting a UPR as a consequence of HLA-B27 up-regulation, where it correlates with activation of XBP-1 splicing. Together these findings indicate that the cellular response to endogenous 'danger' that disrupts ER homeostasis is coupled to IFN-beta induction by XBP-1, which has implications for the immune response and the pathogenesis of diseases involving the UPR. PMID- 18412160 TI - Protective anti-mycobacterial T cell responses through exquisite in vivo activation of vaccine-targeted dendritic cells. AB - Vaccine efficacy largely depends upon DC targeting and activation. The most potent TLR soluble ligands induce diffuse DC activation, which may be associated with marked pro-inflammatory responses and possibly adverse effects. This raises the concern that effective vaccine adjuvants may similarly rely on widespread DC activation. Using a promising candidate vaccine against tuberculosis (fusion protein of Ag85B and 6-kDa early secretory antigenic target (ESAT-6)) formulated in the potent IC31 adjuvant, DC targeting and activation was studied in vivo, following the fate of antigen and adjuvant in the draining lymph nodes, to define the magnitude of DC targeting/activation required in vivo to induce protective vaccine responses. Unexpectedly, protective IFN-gamma-mediated Ag85B-ESAT-6/IC31 responses were associated to the activation of a minute population (less than 0.3%) of CD11c(+) lymph node DC, without detectable systemic pro-inflammatory responses. This activated peripheral tissue-derived DC population, characterized by enhanced CD80, CD86, CD40 and IL-12p40 expression, was only identified when focusing on adjuvant- or antigen-labeled CD11c(+) DC, which were found to support T cell proliferation. Immunization with aluminum hydroxide adjuvant (Alum) resulted in a similar proportion of antigen-associated DC but without detectable enhancement of CD80, CD86, CD40 or IL-12p40 expression. Thus, potent protective IFN-gamma-producing responses may be elicited by the exquisite activation of a minute number of in vivo targeted DC. PMID- 18412161 TI - Immunosenescent colitogenic CD4(+) T cells convert to regulatory cells and suppress colitis. AB - Inflammatory bowel diseases progress steadily by the expansion of colitogenic CD4(+) cells. However, it remains unknown whether colitogenic CD4(+) cells are long-living like memory cells or exhausted like effector cells. To assess the longevity of colitogenic lamina propria (LP) CD4(+) cells, we performed sequential transfers of LP CD4(+) cells from colitic CD4(+)CD45RB(high) cell transferred SCID mice into new SCID mice. Although SCID mice transferred with colitic LP CD4(+) cells stably developed colitis until at least the sixth transfer, the interval to the development of colitis gradually lengthened as the number of transfers increased. The incidence of colitis gradually decreased after the seventh transfer. Furthermore, non-colitic LP CD4(+) cells from mice transferred over seven times expressed significantly higher levels of PD-1 and produced significantly lower amounts of IFN-gamma, TNF-alpha, and IL-17 than colitic LP CD4(+) cells recovered after the first transfer. Most notably, we found that re-transfer of non-colitic LP CD4(+) cells recovered after multiple transfers prevented the development of colitis in SCID mice co-transferred with CD4(+)CD45RB(high) cells. Thus, colitogenic LP CD4(+) cells may be exhausted over time, become non-functional, convert to regulatory cells, and finally suppress colitis in the process of immunosenescence. PMID- 18412162 TI - Intercellular MHC transfer between thymic epithelial and dendritic cells. AB - Thymic dendritic cells (DC) and epithelial cells play a major role in central tolerance but their respective roles are still controversial. Epithelial cells have the unique ability to ectopically express peripheral tissue-restricted antigens conferring self-tolerance to tissues. Paradoxically, while negative selection seems to occur for some of these antigens, epithelial cells, contrary to DC, are poor negative selectors. Using a thymic epithelial cell line, we show the functional intercellular transfer of membrane material, including MHC molecules, occurring between epithelial cells. Using somatic and bone marrow chimeras, we show that this transfer occurs efficiently in vivo between epithelial cells and, in a polarized fashion, from epithelial to DC. This novel mode of transfer of MHC-associated, epithelial cell-derived self-antigens onto DC might participate to the process of negative selection in the thymic medulla. PMID- 18412163 TI - C-reactive protein specifically binds to Fcgamma receptor type I on a macrophage like cell line. AB - C-reactive protein (CRP) is a prototype acute-phase protein that may be intimately involved in human disease. Its cellular receptors are still under debate; the main candidates are FcR for immunoglobulin G, as CRP was shown to bind specifically to FcgammaRI and FcgammaRIIa. Using ultrasensitive confocal live-cell imaging, we have studied CRP binding to FcgammaR naturally expressed in the plasma membranes of cells from a human leukemia cell line (Mono Mac 6). These macrophage-like cells express high levels of FcgammaRI and FcgammaRII. They were shown to bind fluorescently labeled CRP with micromolar affinity, KD = (6.6 +/- 1.5) microM. CRP binding could be inhibited by pre-incubation with human but not mouse IgG and was thus FcgammaR-specific. Blocking of FcgammaRI by an FcgammaRI specific antibody abolished CRP binding essentially completely, whereas application of antibodies against FcgammaRII did not have a noticeable effect. In fluorescence images of Mono Mac 6 cells, the intensity patterns of bound CRP were correlated with those of FcgammaRI, but not FcgammaRII. These results provide clear evidence of specific interactions between CRP and FcgammaR (predominantly FcgammaRI) naturally expressed on macrophage-like cells. PMID- 18412164 TI - Induction of HIV-specific T and B cell responses with a replicating and conditionally infectious lentiviral vaccine. AB - The development of an HIV vaccine that induces broad and potent immunity is critically needed. Viruses, including lentiviruses, have been used as vectors for ex vivo transduction of antigens into dendritic cells (DC). We hypothesized that DC transduced with a vector that allows selective infection of DC could induce potent immunity by continually priming DC. A lentiviral vector encoding HIV gag pol without env would form viral cores in transduced DC, but would release non infectious particles by budding into endosomes and releasing apoptotic bodies or exosomes containing viral cores. DC function by endocytosing DC-derived apoptotic bodies, and they are specialized in their ability to move endocytic contents into the cytoplasm. We postulated that endocytosis of vector cores could lead to transduction of a second round of DC. In this report, we demonstrate accumulation of viral cores inside transduced DC and show second-round transduction of immature DC that endocytose transduced DC in vitro. The effectiveness of immunization of mice with transduced DC to induce specific lymphocyte activation was assessed. Mice developed antigen-specific T cell responses and specific antibodies after immunization. Transduction of DC with a replication-competent but conditionally infectious lentivirus could be a novel vaccine strategy for HIV. PMID- 18412167 TI - Pd-catalyzed cross-coupling reactions with carbonyls: application in a very efficient synthesis of 4-aryltetrahydropyridines. PMID- 18412165 TI - Epstein-Barr virus-induced gene 3 negatively regulates IL-17, IL-22 and RORgamma t. AB - Epstein-Barr virus-induced gene 3 (EBI3) associates with p28 to form IL-27 and with IL-12p35 to form IL-35. IL-27Ralpha(-/-) mice studies indicate that IL-27 negatively regulates Th17 cell differentiation. However, no EBI3, p28 or p35 deficiency studies that directly address the role of EBI3, p28 or p35 on Th17 cells have been done. Here, we demonstrate that spleen cells derived from EBI3(-/ ) mice produce significantly higher levels of IL-17 as well as IL-22 upon stimulation with OVA. In vitro derived EBI3(-/-) Th17 cells also produced significantly higher levels of IL-17 and IL-22 than WT cells. The frequency of IL 17-producing cells was also elevated when EBI3(-/-) cells were cultured under Th17 conditions. In addition, spleen cells from EBI3(-/-) mice immunized with Listeria monocytogenes produced significantly elevated levels of IL-17 and IL-22. Furthermore, the Th17 transcription factor RORgamma t was significantly enhanced in EBI3(-/-) cells. Finally, EBI3(-/-) mice exhibited a reduced bacterial load following an acute challenge with L. monocytogenes or a re-challenge of previously immunized mice, suggesting that EBI3 negatively regulates both innate and adaptive immunity. Taken together, these data provide direct evidence that EBI3 negatively regulates the expression of IL-17, IL-22 and RORgamma t as well as protective immunity against L. monocytogenes. PMID- 18412166 TI - Role of PI3Kdelta and PI3Kgamma in inflammatory arthritis and tissue localization of neutrophils. AB - The p110delta isoform of class I phosphoinositide 3-kinase (PI3K) plays a major role in B cell receptor signaling, while its p110gamma counterpart is thought to predominate in leukocyte chemotaxis. Consequently, emphasis has been placed on developing PI3Kgamma selective inhibitors to treat disease states that result from inappropriate tissue accumulation of leukocytes. We now demonstrate that PI3Kdelta blockade is effective in treating an autoimmune disorder in which neutrophil infiltration is required for tissue injury. Using the K/BxN serum transfer model of arthritis, in which neutrophils and leukotriene B(4) (LTB(4)) participate, we show that genetic deletion or selective inhibition of PI3Kdelta diminishes joint erosion to a level comparable to its PI3Kgamma counterpart. Moreover, the induction and progression of joint destruction was profoundly reduced in the absence of both PI3K isoforms and correlated with a limited ability of neutrophils to migrate into tissue in response to LTB(4). However, the dynamic interplay between these isoforms is not pervasive, as fMLP-induced neutrophil extravasation was primarily reliant on PI3Kgamma. Our results not only demonstrate that blockade of PI3Kdelta has potential therapeutic value in the treatment of chronic inflammatory conditions, but also provide evidence that dual inhibition of these lipid kinases may yield superior clinical results. PMID- 18412168 TI - Quaternary stereogenic carbon atoms in complex molecules by an asymmetric, organocatalytic, triple-cascade reaction. PMID- 18412169 TI - Oligodendrocyte-specific protein is encephalitogenic in rhesus macaques and induces specific demyelination of the optic nerve. AB - Oligodendrocyte-specific protein (OSP) is a candidate autoantigen in the development of multiple sclerosis (MS). We evaluated the potential of OSP to induce EAE in rhesus monkeys, an out bred animal model for MS that is immunologically close to humans. Since OSP is a four-membrane spanning protein with highly hydrophobic regions, we synthesized recombinant proteins encompassing only the hydrophilic regions of human OSP (soluble (s)hOSP). Immunization with shOSP proteins induced clinical signs and histological features of optic neuritis in four out of ten rhesus monkeys. The development of clinical disease was associated with the presence of a strong cellular proliferative response to the immunizing shOSP protein. Analysis of the cellular responses in combination with neuropathological observations also indicates an important role for neutrophils in the disease process. Interestingly, all immunized monkeys developed antibody responses to OSP peptide 103-123, a B cell epitope previously identified in MS patients. These responses did not correlate with the development of clinical disease, but may have relevance as a biomarker for immunoreactivity towards OSP in myelin disorders. Our data demonstrate that in rhesus monkeys immune responses directed at OSP are encephalitogenic, leading to inflammatory responses throughout the central nervous system and to selective demyelination of the optic nerve. PMID- 18412170 TI - Degenerate TCR recognition and dual DR2 restriction of autoreactive T cells: implications for the initiation of the autoimmune response in multiple sclerosis. AB - TCR degeneracy may facilitate self-reactive T cell activation and the initiation of an autoimmune response in multiple sclerosis (MS). MHC class II alleles of the DR2 haplotype DR2a (DRB5*0101) and DR2b (DRB1*1501) are associated with an increased risk for MS in Caucasian populations. In order to selectively expand and characterize T cells with a high degree of TCR degeneracy that recognize peptides in the context of disease-associated DR2 alleles, we developed DR2 anchored peptide mixtures (APM). We report here that DR2-APM have a high stimulatory potency and can selectively expand T cells with a degenerate TCR (TCR(deg)). Due to the low concentration of individual peptides in the mixtures, T cell clones' proliferative response to DR2-APM implies that multiple peptides stimulate the TCR, which is a characteristic of TCR(deg). The frequency of DR2 APM-reactive T cells is significantly higher in MS patients than in healthy controls, suggesting that they may play a role in the development of the autoimmune response in MS. DR2-APM-reactive cells have a dual DR2 restriction: they recognize DR2-APM in the context of both DR2a and DR2b molecules. The DR2 APM-reactive cells' IL-17 secretion, together with cross-reactivity against myelin peptides, may contribute to their role in the development of autoimmune response in MS. PMID- 18412171 TI - Forced overexpression of either of the two common human Foxp3 isoforms can induce regulatory T cells from CD4(+)CD25(-) cells. AB - The forkhead/winged helix transcription factor (Foxp3) is expressed as two different isoforms in humans: the full-length isoform (Foxp3FL) and an alternative-splicing product lacking the exon 2 (Foxp3DeltaE2). We here studied the cellular distribution of Foxp3 isoforms by quantitative PCR and evaluated the functional outcome of retroviral transduction of Foxp3FL and Foxp3DeltaE2 genes into CD4(+)CD25(-) cells. In PBMC, both isoforms were preferentially expressed in CD4(+)CD25(hi) cells. In single-cell-sorted and expanded Treg, both Foxp3 isoforms were expressed simultaneously but without a fixed ratio. Forced expression of Foxp3FL or Foxp3DeltaE2 genes in CD4(+)CD25(-) T cells induced bona fide Treg that not only displayed Treg phenotype but also were anergic and mediated significant suppressive activity against CD3-activated CD4(+)CD25(-) cells. GFP(-) nontransduced cells or cells transduced with an empty vector showed no Treg phenotype, anergy or suppressive activities. In conclusion, our results reveal that both Foxp3 isoforms possess similar capacities to induce Treg; however, unnaturally high expression levels are required to convey Treg functions to CD4(+)CD25(-) cells. As both Foxp3 isoforms appear to be expressed in an independent fashion, studies aiming at quantification of Treg in peripheral blood or in tissue samples can benefit from determination of total Foxp3 levels rather than one of the isoforms. PMID- 18412173 TI - Mexico, the father of the pill and the race for cortisone. PMID- 18412174 TI - Computational chemistry approaches to drug discovery in signal transduction. AB - The advent of therapeutic strategies aimed at targeting specific macromolecular components of deregulated signaling pathways associated with particular disease states has given rise to the idea that it should be possible to design ligands as drug candidates to these targets from first principles. This concept has been beckoning for a long time but structure-based ligand design only became feasible once it was possible to determine the 3-D structures of molecular targets at atomic resolution. However, structure-based design turned out to be difficult, chiefly because under physiological conditions both receptors and ligands are not static but they behave dynamically. While it is possible to design ligands with high steric and electronic complementarity to a receptor site, it is always uncertain how biologically relevant the assumed conformations of both ligand and receptor actually are. The fact that it remains beyond our current abilities to predict with sufficient accuracy the affinity between hypothetical ligand and receptor poses is in part connected with this problem and continues to confound the reliable prediction of drug-like ligands for therapeutic targets. Nevertheless, significant progress has been made and so-called virtual screening methods that use computational methods to dock candidate ligands into receptor sites and to score the resulting complexes are now used routinely as one of the components in drug discovery screening campaigns. Here an overview is given of the underlying principles, implementations, and applications of structure-guided computational design technologies. Although the emphasis is on receptor-based strategies, mention will also be made of some of the more established ligand based approaches, such as similarity analyses and quantitative structure-activity relationship methods. PMID- 18412175 TI - The application of cell-based label-free technology in drug discovery. AB - Cell-based assays are an important part of the drug discovery process allowing for interrogation of targets and pathways in a more physiological setting compared to biochemical assays. One of the main hurdles in the cell-based assay field is to design sufficiently robust assays with adequate signal to noise parameters while maintaining the inherent physiology of the pathway or target being investigated. Conventional label and reporter-based cell assays may be more prone to artifacts due to considerable manipulation of the cell either by the label or over-expression of targets or reporter proteins. Cell-based label-free technologies preclude the need for cellular labeling or over-expression of reporter proteins, utilizing the inherent morphological and adhesive characteristics of the cell as a physiologically relevant and quantitative readout for various cellular assays. Furthermore, these technologies utilize non invasive measurements allowing for time resolution and kinetics in the assay. In this article, we have reviewed the various label-free technologies that are being used in drug discovery settings and have focused our discussion on impedance based label-free technologies and its main applications in drug discovery. PMID- 18412180 TI - Editorial: Novel approaches to drug discovery in signal transduction. PMID- 18412181 TI - Cell-based assays in GPCR drug discovery. In this issue. PMID- 18412184 TI - Sustainable metal catalysis with iron: from rust to a rising star? PMID- 18412188 TI - Carbon-nitrogen bond-forming reactions of dialkyl azodicarboxylate: a promising synthetic strategy. AB - Azodicarboxylates have found applications in electrophilic amination reactions and in pericyclic reactions. The nucleophilic trigger in Mitsunobu reactions, that is, the zwitterion formed from triphenylphosphine and dialkyl azodicarboxylate, has been utilized recently in various heterocyclic constructions. This Focus Review summarizes the potential utility of azodicarboxylates in various carbon-nitrogen bond-forming reactions. PMID- 18412189 TI - Domain-domain interactions in the iterative type I polyketide synthase ATX from Aspergillus terreus. PMID- 18412190 TI - Identifying selective protein tyrosine phosphatase substrates and inhibitors from a fluorogenic, combinatorial peptide library. PMID- 18412191 TI - The mildiomycin biosynthesis: initial steps for sequential generation of 5 hydroxymethylcytidine 5'-monophosphate and 5-hydroxymethylcytosine in Streptoverticillium rimofaciens ZJU5119. AB - Mildiomycin (MIL) is a peptidyl nucleoside antibiotic with strong activity against powdery mildew disease of plants. We have cloned the MIL biosynthetic gene cluster in Streptoverticillum rimofaciens ZJU5119 and shown that this organism also produces the related antifungal compound, deshydroxymethyl mildiomycin (dHM-MIL). A cosmid genomic library was screened for a putative nucleotide hydrolase gene that is related to blsM from the blasticidin S cluster. Six cosmids were identified that contained a 3.5 kb DNA fragment that harbors a homologue of blsM. The sequence of the fragment revealed two open-reading frames that are likely to function in MIL formation: milA is a CMP hydroxymethylase gene and milB is the homologue of the CMP hydrolase gene blsM. Insertional disruption of milA abolished the production of MIL but not dHM-MIL, whereas a milB knockout strain did not produce either of the peptidyl nucleosides. Recombinant MilA was produced in E. coli and shown to specifically introduce a C-5 hydroxymethyl group on CMP, but it did not accept cytosine or dCMP as a substrate. MilB was also expressed and purified from E. coli and shown to efficiently hydrolyze both hydroxymethyl-CMP (HMCMP) and could accept CMP as an alternative substrate. The ratio of free HMC and cytosine released by MilB was ca. 9:1 in in vitro assays, and is consistent with the higher levels of MIL compared to dHM-MIL that are produced by Streptoverticillum rimofaciens. PMID- 18412192 TI - Direct growth of optically stable gold nanorods onto polyelectrolyte multilayered capsules. PMID- 18412193 TI - Synthesis and application of fluorescein- and biotin-labeled molecular probes for the chemokine receptor CXCR4. AB - The design, synthesis, and bioevaluation of fluorescence- and biotin-labeled CXCR4 antagonists are described. The modification of D-Lys8 at an epsilon-amino group in the peptide antagonist Ac-TZ14011 derived from polyphemusin II had no significant influence on the potent binding of the peptide to the CXCR4 receptor. The application of the labeled peptides in flow cytometry and confocal microscopy studies demonstrated the selectivity of their binding to the CXCR4 receptor, but not to CXCR7, which was recently reported to be another receptor for stromal cell derived factor 1 (SDF-1)/CXCL12. PMID- 18412196 TI - Dedication to Professor Hans-Gunter Gassen. On the occasion of his 70th birthday and retirement. PMID- 18412198 TI - Macrocyclic high-spin (S=2) molecule: spin identification of a sterically rigid metacyclophane-based nitroxide tetraradical by two-dimensional electron spin transient nutation spectroscopy. PMID- 18412199 TI - Is deuterium always smaller than protium? PMID- 18412201 TI - The molecular cluster [Bi10Au2](SbBi3Br9)2. PMID- 18412200 TI - Benzopyrenomycin, a cytotoxic bacterial polyketide metabolite with a benzo[a]pyrene-type carbocyclic ring system. PMID- 18412202 TI - Simultaneous copper(I)-catalyzed azide-alkyne cycloaddition (CuAAC) and living radical polymerization. PMID- 18412203 TI - The "complex-in-a-complex" cations [(acac)2M subset Ru6(p iPrC6H4Me)6(tpt)2(dhbq)3]6+: A trojan horse for cancer cells. PMID- 18412204 TI - Heterobimetallic cooperation mediates the transformation of white phosphorus into zwitterionic catena-phosphonium(+)diphosphenide(-) ligands. PMID- 18412205 TI - A carbonyl ylide cycloaddition approach to platensimycin. PMID- 18412206 TI - The low basicity of phosphabenzenes: first examples of protonation, alkylation, and silylation reactions. PMID- 18412207 TI - Self-assembly of phthalocyanine nanotubes by vapor-phase transport. PMID- 18412208 TI - Regioselective H/D exchange at the side-chain of ethylbenzene on dealuminated zeolite H-Y studied by in situ MAS NMR-UV/Vis spectroscopy. PMID- 18412209 TI - Amyloids: not only pathological agents but also ordered nanomaterials. AB - Amyloid fibers constitute one of the most abundant and important naturally occurring self-associated assemblies. A variety of protein and peptide molecules with various amino acid sequences form these highly stable and well-organized assemblies under diverse conditions. These assemblies display phase states ranging from liquid crystals to rigid nanotubes. The potential applications of these supramolecular assemblies exceed those of synthetic polymers since the building blocks may introduce biological function in addition to mechanical properties. Here we review the structural characteristics of amyloidal supramolecular assemblies, their potential use as either natural or de novo designed sequences, and the range of applications that have been demonstrated so far. PMID- 18412210 TI - Growing molecular crystals on inorganic crystals: polar structure induced by noncentrosymmetric templates. PMID- 18412211 TI - Clockwork PCR including sample preparation. PMID- 18412212 TI - Two-phase (bio)catalytic reactions in a table-top centrifugal contact separator. PMID- 18412213 TI - A highly selective luminescent switch-on probe for histidine/histidine-rich proteins and its application in protein staining. PMID- 18412214 TI - The cation-anion interaction in ionic liquids probed by far-infrared spectroscopy. PMID- 18412215 TI - Substituent effects, reactant preorganization, and ligand exchange control the reactivity in Rh(I)-catalyzed (5+2) cycloadditions between vinylcyclopropanes and alkynes. PMID- 18412218 TI - A novel method for the detection of viable human pancreatic beta cells by flow cytometry using fluorophores that selectively detect labile zinc, mitochondrial membrane potential and protein thiols. AB - Improvement over current methods of beta cell viability assessment is highly warranted in order to efficiently predict the viability and function of beta cells prior to transplantation into type 1 diabetes patients. Dispersed human islet cells were stained with the cell-permeable zinc-selective dye, FluoZin-3 AM, along with the mitochondrial membrane potential indicator [(tetramethylrhodamine ethylester (TMRE)] and the thiol-binding dye, monochlorobimane (mBcl), and analyzed by flow cytometry. Islets were subjected to various experimental conditions to validate the usefulness of this method to accurately determine the viability and function of beta cells. Staining with FluoZin-3 revealed the presence of higher amounts of chelatable zinc ions in beta cells than in lymphoid cells and fibroblasts. An intracellular zinc chelator competitively inhibited the binding of FluoZin-3 to zinc ions. Mitochondrial depolarization or oxidative stress minimally affected the binding of mBcl and FluoZin-3, respectively, to thiols and zinc ions. The combination of FluoZin-3, TMRE, and mBcl was sufficient and necessary for the determination of the viability and function of beta cells. The data demonstrate the usefulness of the zinc-specific dye and the indicators of mitochondrial function and thiol levels, to accurately estimate the beta cell viability and function. This novel flow cytometry method has implications for islet transplantation in type 1 diabetes patients. PMID- 18412216 TI - Basophil activation test by flow cytometry: present and future applications in allergology. AB - The diagnosis of allergic reactions in clinical practice rests upon both clinical history and the demonstration of specific immunoglobulin E (sIgE), either in the serum or via skin tests. However, for various reasons, identification of the offending allergen(s) is not always possible. Moreover, not all allergies are IgE mediated. In an attempt to find reliable methods to investigate hypersensitivity reactions, histamine and sulfidoleukotriene release tests have long been introduced. However, relatively few comprehensive quality reports have been published so far. Upon challenge with a specific allergen, basophils not only secrete quantifiable bioactive mediators but also upregulate the expression of different markers which can be detected efficiently by flow cytometry using specific monoclonal antibodies. This review addresses the principals, particular technical aspects and pitfalls as well as the clinical and research applications of flow-assisted analysis of in vitro activated basophils. PMID- 18412219 TI - Retinoic acid-induced inner ear teratogenesis caused by defective Fgf3/Fgf10 dependent Dlx5 signaling. AB - BACKGROUND: Retinoic acid (RA) is essential for inner ear development. However, exposure to excess RA at a critical period leads to inner ear defects. These defects are associated with disruption in epithelial-mesenchymal interactions. METHODS: This study investigates the role of Dlx5 in the epithelial-mesenchymal interactions that guide otic capsule chondrogenesis, as well as the effect of excess in utero RA exposure on Dlx5 expression in the developing mouse inner ear. Control of Dlx5 by Fgf3 and Fgf10 under excess RA conditions is investigated by examining the developmental window during which Fgf3 and Fgf10 are altered by in utero RA exposure and by testing the ability of Fgf3 and Fgf10 to mitigate the reduction in chondrogenesis and Dlx5 expression mediated by RA in high-density cultures of periotic mesenchyme containing otic epithelium, a model of epithelial mesenchymal interactions in which chondrogenic differentiation of periotic mesenchyme ensues in response to induction by otic epithelium. RESULTS: Dlx5 deletion alters expression of TGFbeta(1), important for otic capsule chondrogenesis, in the developing inner ear and compromises the ability of cultured periotic mesenchyme containing otic epithelium, harvested from Dlx5 null embryos, to differentiate into cartilage when compared with control cultures. Downregulation in Dlx5 ensues as a consequence of in utero RA exposure in association with inner ear dysmorphogenesis. This change in Dlx5 is noted at embryonic day 10.5 (E10.5), but not at E9.5, suggesting that Dlx5 is not a direct RA target. Before Dlx5 downregulation, Fgf3 and Fgf10 expression is modified in the inner ear by excess RA, with the ability of exogenous Fgf3 and Fgf10 to rescue chondrogenesis and Dlx5 expression in RA-treated cultures of periotic mesenchyme containing otic epithelium supporting these fibroblast growth factors (FGFs) as intermediary genes by which RA mediates its effects. CONCLUSIONS: Disruption in an Fgf3, -10/Dlx5 signaling cascade is operant in molecular mechanisms of inner ear teratogenesis by excess RA. PMID- 18412220 TI - Phosphorylation of neurofilament subunit NF-M is regulated by activation of NMDA receptors and modulates cytoskeleton stability and neuronal shape. AB - The cytoskeleton is essential for the structural organization of neurons and is influenced during development by excitatory stimuli such as activation of glutamate receptors. In particular, NMDA receptors are known to modulate the function of several cytoskeletal proteins and to influence cell morphology, but the underlying molecular and cellular mechanisms remain unclear. Here, we characterized the neurofilament subunit NF-M in cultures of developing mouse cortical neurons chronically exposed to NMDA receptor antagonists. Western blots analysis showed that treatment of cortical neurons with MK801 or AP5 shifted the size of NF-M towards higher molecular weights. Dephosphorylation assay revealed that this increased size of NF-M observed after chronic exposure to NMDA receptor antagonists was due to phosphorylation. Neurons treated with cyclosporin, an inhibitor of the Ca(2+)-dependent phosphatase calcineurin, also showed increased levels of phosphorylated NF-M. Moreover, analysis of neurofilament stability revealed that the phosphorylation of NF-M, resulting from NMDA receptor inhibition, enhanced the solubility of NF-M. Finally, cortical neurons cultured in the presence of the NMDA receptor antagonists MK801 and AP5 grew longer neurites. Together, these data indicate that a blockade of NMDA receptors during development of cortical neurons increases the phosphorylation state and the solubility of NF-M, thereby favoring neurite outgrowth. This also underlines that dynamics of the neurofilament and microtubule cytoskeleton is fundamental for growth processes. PMID- 18412221 TI - Accidental Nd:YAG laser-induced choroidal neovascularization. AB - BACKGROUND: There are few reports in the literature to describe the natural history of an occupational Nd:YAG laser-induced retinal injury. METHODS: The chronological response to a Nd:YAG laser-induced injury was studied in a 48-year old male using the techniques of color fundus photography, fluorescein angiography and optical coherence tomography. When he developed choroidal neovascularization his response to photodynamic therapy with verteporfin was recorded. RESULTS: The patient initially developed a macular hole, which spontaneously closed. Three months after the injury a choroidal neovascular membrane developed. This stabilized after the use of photodynamic therapy. CONCLUSIONS: Surgical closure of Nd:YAG induced macular holes should be delayed to allow spontaneous closure. Photodynamic therapy was successful in treating choroidal neovascularization in this case. PMID- 18412222 TI - High-resolution two-photon excitation microscopy of ocular tissues in porcine eye. AB - BACKGROUND AND OBJECTIVE: Two-photon excitation laser scanning microscopy (TPM), based on nonlinear optical (NLO) response under high irradiance, is currently being extensively employed for diagnostic purposes in biomedical fields and becomes more and more an interesting imaging technique in the intact bulk tissue examination. In this study, this nonlinear-excitation imaging technique including two-photon-mediated autofluorescence (2PF) and second harmonic generation (SHG) was employed to investigate the microstructures in the whole-mount scleral, retinal, and corneal tissues of porcine eyes with intracellular spatial resolution and high signal-to-noise ratio. MATERIALS AND METHODS: Image acquisition was based on the intense 80 MHz femtosecond (fs) near-infrared (NIR) laser pulses, emitted from a mode-locked solid-state titanium:sapphire system. By integrating a high-numerical aperture diffraction-limited objective, the whole mount ocular specimens could be viewed from the surface of eye globes further to a 200 microm depth. Under high light irradiance at the order of MW-GW/cm2, more than one photon was simultaneously absorbed by endogenous molecules in ocular tissues. RESULTS: The cellular and fibrous components of whole-mount scleral and corneal tissues were selectively displayed in situ by in-tandem detection of 2PF and SHG with high efficiency without the assistance of any exogenous dye. NLO images of fibroblasts and mature elastic fibers in sclerae as well as of the retina radial Muller glial cells, ganglion cells, bipolar cells, photoreceptors, and retina pigment epithelial (RPE) cells were acquired with subcellular spatial resolution. In particular, the microstructural topography of cells and extracellular components in the whole-mount ocular tissues was elucidated in situ. CONCLUSION AND OUTLOOK: The combination of the sensitive image acquisition technique allows to selectively studying of three-dimensional (3-D) architecture of cellular microstructures and extracellular matrix arrangement in situ at substantial depths in bulk tissues. The data obtained provided the primary knowledge for further studies of imaging entire eye globes based on two-photon excitation microscopy. PMID- 18412223 TI - Laser induced fragmentation of salivary stones: an in vitro comparison of two different, clinically approved laser systems. AB - BACKGROUND: Clinical laser lithotripsy in urology promises a good fragmentation combined with a minimal risk of soft tissue damage and low medical complications. This in vitro study investigates the fragmentation of salivary stones by means of two clinically used laser systems. MATERIALS AND METHODS: The effects induced by the FREDDY laser (WOM, Germany, lambda = 532 nm/1,064 nm, E(pulse) = 120-160 mJ/pulse) and the Ho:YAG (AURIGA, StarMedTec, Germany, lambda = 2,100 nm, E(pulse) = 300-800 mJ/pulse) on clinical salivary calculi (n = 15) and on salivary gland tissue were investigated using clinical laser parameter settings. All experiments were performed in an under water experimental set-up using flexible fibres (core diameter 230 microm) positioned in front of each specimen. In order to assess fragmentation efficacy, each stone was placed on a grating (rhombic mash-diameter 1-3 mm). The fragmentation rate was calculated with respect to the energy applied (mg/J), to the number of pulses (mg/pulse), and to the time needed (mg/minute). In addition the composition of the stones were analysed spectrographically. The soft tissue interaction on human salivary duct mucosa was examined histologically (HE-staining). RESULTS: Spectrographic composition of the salivary stones showed a two component ratio of protein/carbonate apatite varying between 5/95 and 25/75. Stones treated by the Ho:YAG were vaporised in a milling-like process, while using the FREDDY laser stones are cracked into pieces and fragmentation failed in two cases. The fragmentation rates achieved by the FREDDY laser were greater than those of the Ho:YAG laser, but fragments mainly bigger. A dependency on the composition of the stones could not be found. Laser pulse effects on soft tissue were found slightly beyond the mucosa. CONCLUSION: This study clearly demonstrated the different processes of destroying salivary stones using two different laser systems. While the Ho:YAG vaporises the calculi in a more milling and soft sense, the FREDDY shows a more cracking and explosive destruction. Although both laser systems showed little direct risk to the surrounding tissue, it has to be proven whether cracked and accelerated particles could cause harm to soft tissue. With respect to this, further in vitro studies and clinical treatments in selected cases are needed to proof these results. PMID- 18412224 TI - Photodynamic treatment as a novel approach in the therapy of arthritic joints. AB - BACKGROUND AND OBJECTIVE: Minimal invasive local treatment of joints is a desirable option in the therapy of rheumatoid arthritis (RA). Aim of this study was to evaluate the effects of photodynamic treatment (PDT) with different doses of the photosensitizer meta-tetra(hydroxyphenyl)chlorin (m-THPC; or temoporfin) in a murine model of RA (antigen-induced arthritis, AIA). METHODS IN VIVO DISTRIBUTION: The distribution of native and liposomal m-THPC (including a formulation with polyethylene glycol [PEG] coating) was assessed by fluorescence spectrometry in arthritic joints, normal joints, and skin. TREATMENT: AIA mice received different concentrations of pegylated liposomal m-THPC (0.1, 0.05, 0.01, or 0.005 mg/kg body weight; n = 5 per group) and subjected to PDT with a laser system 12 hours post-injection of the photosensitizer. Treatment effects were evaluated histologically in comparison to untreated AIA (n = 5). RESULTS: Pegylated liposomal m-THPC showed the most favorable accumulation in arthritic joints compared to native m-THPC and to non-peg-liposomal m-THPC, therefore it was selected as photosensitizer for PDT treatment. In comparison to untreated AIA, PDT reduced the arthritic score with all doses of pegylated liposomal m THPC; statistical significant effects were obtained with doses of 0.05 and 0.01 mg/kg. CONCLUSION: Our study demonstrated that local PDT of arthritic joints is feasible. Application of pegylated liposomal m-THPC for PDT resulted in significant reduction of arthritis scores. PMID- 18412226 TI - Transient immunoreactivity after laser tattoo removal: report of two cases. AB - BACKGROUND AND OBJECTIVE: Laser tattoo removal is one of most commonly used indications for medical lasers. Professional tattoos contain a multitude of potentially immunogenic chemicals that are released or modified by laser treatment. We studied potential immunologic reactions following laser tattoo removal. STUDY DESIGN/PATIENTS AND METHODS: Case report of two patients with immunologic reactions after laser tattoo removal. RESULTS: Two patients developed transient immunoreactivity that presented as regional lymphadenopathy after laser tattoo removal of professional black and blue-green tattoos. These reactions resolved without any complications. CONCLUSIONS: Tattoo pigments released or modified by laser therapy may trigger transient immunoreactivity in susceptible individuals. PMID- 18412227 TI - Rosacea treatment using the new-generation, high-energy, 595 nm, long pulse duration pulsed-dye laser. AB - BACKGROUND AND OBJECTIVES: Rosacea results from sun-induced hyper-vascularity of exposed facial skin, often accompanied by acneiform papules and pustules. The pulsed-dye laser has an unparalleled safety record and emits an ideal wavelength for treating the abnormal vessels that occur in rosacea patients. In this study we investigate the ability of the long pulse-duration pulsed-dye laser to improve rosacea. STUDY DESIGN/MATERIALS AND METHODS: Twenty subjects with rosacea were treated with the high-energy, long pulse-duration pulsed-dye laser. An elliptical spot and long pulse-duration was used to spot-treat linear telangiectasias, while a circular spot and short pulse-duration was used to treat the entire face. Improvement was determined by blinded evaluation of photographs and by the treating physician's subjective evaluation, before and 8 weeks following the final treatment. RESULTS: The average rosacea score as estimated by the treating physician decreased from 2.7+/-1.1 to 1.4+ 0.7 (mean+SD) on a 0-6 scale with 0 representing no rosacea and 6 representing the most severe rosacea (P<0.001 level). The average rosacea score as rated by blinded physician observers scoring digital photos was 2.3+/-1.3 before treatment and 1.4+/-0.9 8 weeks following treatment, using a 0 (mild) to 6 (severe) scale. CONCLUSIONS: The high-energy, long pulse-duration pulsed-dye laser improves rosacea with a very favorable safety profile, and less purpura than resulted from earlier generation pulsed-dye lasers. PMID- 18412228 TI - Scattered UV irradiation during VISX excimer laser keratorefractive surgery. AB - PURPOSE: To evaluate the potential occupational health hazards associated with scattered ultraviolet (UV) radiation during photorefractive keratectomy (PRK) using the VISX Star S3 excimer laser. SETTING: The Laser Vision Center, National Naval Medical Center, Bethesda, Maryland, USA. METHODS: Intraoperative radiometric measurements were made with the Ophir Power/Energy Meter (LaserStar Model PD-10 with silicon detector) during PRK treatments as well as during required calibration procedures at a distance of 20.3 cm from the left cornea. These measurements were evaluated using a worst-case scenario for exposure, and then compared with the American Conference of Governmental Industrial Hygeinists (ACGIH) Threshold Value Limits (TVL) to perform a risk/hazard analysis. RESULTS: During the PRK procedures, the highest measured value was 248.4 nJ/pulse. During the calibration procedures, the highest measured UV scattered radiation level was 149.6 nJ/pulse. The maximum treatment time was 52 seconds. Using a worst-case scenario in which all treatments used the maximum power and time, the total energy per eye treated was 0.132 mJ/cm2 and the total UV radiation at close range (80 cm from the treated eye) was 0.0085 mJ/cm2. With a workload of 20 patients, the total occupational exposure at 80 cm to actinic UV radiation in an 8-hour period would be 0.425 mJ/cm2. CONCLUSIONS: The scattered actinic UV laser radiation from the VISX Star S3 excimer laser did not exceed occupational exposure limits during a busy 8-hour workday, provided that operating room personnel were at least 80 cm from the treated eye. While the use of protective eyewear is always prudent, this study demonstrates that the trace amounts of scattered laser emissions produced by this laser do not pose a serious health risk even without the use of protective eyewear. PMID- 18412229 TI - Multiphoton excitation fluorescence microscopy of 5-aminolevulinic acid induced fluorescence in experimental gliomas. AB - BACKGROUND AND OBJECTIVE: The clinical usefulness of 5-ALA guided detection of tumor tissue has been demonstrated for a number of malignancies. However, current techniques of intraoperative detection of protoporphyrin IX fluorescence in situ do not offer subcellular resolution. Therefore, discrimination of non-specific 5 ALA induced fluorescence remains difficult. MATERIALS AND METHODS: In this study we have used an orthotopic glioma model to analyze PpIX fluorescence in tumor tissue and normal brain by multiphoton excitation microscopy after intraperitoneal administration of 5-ALA. A DermaInspect in vivo imaging system was used for autofluorescence measurements at 750 nm excitation and detection in the green channel of a standard photomultiplier module. For detection of PpIX fluorescence at different excitation wavelengths a red sensitive version of the photomultiplier and a filter combination of short pass filters and a color glass long pass filter was used restricting the sensitivity in the red channel to a range of 580-700 nm. RESULTS: Multiphoton microscopy allowed a higher structural definition of tumor tissue based on the excitation of 5-ALA induced PpIX fluorescence compared to autofluorescence imaging. The high resolution of multiphoton microscopy allowed discrimination of fluorescence from the cytoplasm of tumor cells and 5-ALA induced PpIX fluorescence of normal brain parenchyma adjacent to tumor. Fluorescence lifetime imaging showed significantly longer fluorescence lifetimes of 5-ALA induced PpIX fluorescence in tumor tissue compared to normal brain. This allowed definition and visualization of the tumor/brain interface based on this parameter alone. CONCLUSION: Multiphoton microscopy of 5-ALA induced PpIX fluorescence in brain tumor tissue conceptually provides a high resolution diagnostic tool, which in addition to structural information may also provide photochemical/functional information. PMID- 18412230 TI - In vitro resistance of articular chondrocytes to 5-Aminolevulinic acid based photodynamic therapy. AB - OBJECTIVE: 5-Aminolevulinic acid based photodynamic therapy (5-ALA-PDT) has revealed promising results in the treatment of inflammatory joint diseases due to the sensitivity of inflamed synovial tissue. For 5-ALA-PDT to be safe and beneficial for intra-articular applications, resistance of chondrocytes is essential to prevent cartilage damage. As no data yet exist, the aim of the present study was to assess in vitro the response of the chondrocytes to 5-ALA PDT and to compare with osteoblasts and synovial tissue derived cells. METHODS: Bovine articular chondrocytes, osteoblasts, and synovial cells were subjected to 5-ALA-PDT in cell culture. The PpIX accumulation and the function of the cells were assessed for up to 12 days. RESULTS: Bovine chondrocytes showed lower PpIX fluorescence upon incubation with 5-ALA (0.0-2.0 mM) for 4 hours as compared to osteoblasts and synovial cells suggesting a low PpIX accumulation. After incubation with 0.5 mM 5-ALA and application of light at a dose of 20 J/cm2, chondrocytes were functionally not affected (collagen type II and aggrecan mRNA, glycosaminoglycan synthesis) whereas a decrease in the proportion of viable cells was observed in osteoblasts and synovial cells (2+/-2% and 14+/-8%, respectively; chondrocytes 91+/-13%). Chondrocytes showed a 58% reduction of 5-ALA uptake using [3H]5-ALA as compared to osteoblasts and a lower mitochondrial content as assessed by the activity of the mitochondrial marker enzyme citrate synthase (9.2+/- 3.6 mU/mg protein) than osteoblasts (32.6+/-10.5 mU/mg) and synovial cells (60.0+/-10.8 mU/mg). The reduced uptake of 5-ALA and/or the low mitochondrial content, an adaptation to their in vivo environment and the site of PpIX synthesis, presumably explains the lower PpIX content in chondrocytes and their resistance against 5-ALA-PDT. CONCLUSION: 5-ALA-PDT might represent a treatment strategy in inflammatory joint diseases without endangering the cartilage function. However, further in vitro and in vivo experiments are required to confirm this data in the authentic environment of chondrocytes, the articular cartilage. PMID- 18412232 TI - Association between X-linked lissencephaly with ambiguous genitalia syndrome and lenticulostriate vasculopathy in neonate. AB - X-linked lissencephaly with ambiguous genitalia syndrome (XLAG) (OMIM #3000215) is a rare, severe malformation of the brain cortex with abnormal neuronal migration caused by mutations of the ARX gene. All the reported patients with lissencephaly are males who presented with a posterior-to-anterior gradient, moderately increased thickness of the brain cortex, agenesis of corpus callosum, micropenis, and cryptorchidism. We describe the neurosonographic findings associated with the XLAG syndrome. To our knowledge, the association between XLAG and lenticulostriate vasculopathy has not been reported before. PMID- 18412233 TI - Sonographic findings of uterine rupture with expulsion of the fetus into broad ligament. AB - We report the sonographic findings of a rare case of uterine rupture with extrusion of the fetus into the broad ligament during a second-trimester abortion. Sonography revealed the empty uterus with an indistinct defect on the side wall and the dead fetus lying outside, surrounded by a thin membrane. At surgery, the uterine rupture was confirmed with the fetus lying in the broad ligament. This study shows the importance of timely sonography in second trimester abortion, enabling immediate management and preventing further complications. PMID- 18412234 TI - Two years experience of a simple technique of precise ostial coronary stenting. AB - OBJECTIVE: To evaluate a new technique of precise ostial coronary stenting without relying solely on angiography. BACKGROUND: Precise stent positioning at ostial coronary stenosis is difficult because angiography may not be able to profile the coronary ostium, due to vessel overlap and/or foreshortening. This problem is compounded by bobbing or to and fro movement of the stent with cardiac contraction. METHODS: A new technique of precise ostial stenting not dependant on angiography was utilized. A guidewire in a side branch was threaded through the most proximal stent cell and the stent was advanced into the target vessel until it was stopped at the carina. The stent was deployed and the side guidewire withdrawn. All results were documented by intravascular ultrasound (IVUS). RESULTS: From October 2005 to October 2007, 58 patients with significant ostial coronary stenosis required stenting. Seventeen patients were treated in the conventional manner and the remaining 41 patients with the new technique. The ostial locations included 8 left main, 25 left anterior descending, 3 circumflex, 1 obtuse marginal, 3 right coronary, and 1 posterior descending artery. Success, as confirmed by IVUS, was achieved in 40 patients (97.6%). Failure occurred in a right coronary ostial stenosis, which was subsequently treated by the conventional method. There were no complications. CONCLUSION: This new technique is highly successful in cases of difficulty in stenting ostial stenosis guided solely by conventional angiography. PMID- 18412235 TI - Self-expanding and balloon expandable covered stents in the treatment of aortic coarctation with or without aneurysm formation. AB - OBJECTIVES: We describe our complete experience with covered stent implantation for aortic coarctation including short- to medium-term outcomes. BACKGROUND: Coarctation of the aorta is a heterogeneous disease process with multiple associated complications both with and without treatment. Covered stents have evolved to provide greater support to the aortic wall and a varied approach with choice of stent tailored to the anatomy of the patient is required. METHODS: We retrospectively analyzed consecutive patient data from two congenital heart centers from March 2003 to October 2007. RESULTS: We implanted 38 covered stents in 37 patients (20 female) of which three were self-expanding stents. Median age was 29.6 years (9-65) with median weight of 71.5 kg (35-95). The indications for stent placement were native coarctation (n = 13); recurrent coarctation following surgical treatment (n = 11); aneurysm associated with previous coarctation surgery (n = 7); aorto-bronchial fistula leading to acute hemoptysis (n = 2); stent fracture (n = 1); associated arterial duct (n = 2). One patient had a combination of acquired coarctation and false aneurysm. The median screening time was 10 min (3-22). The median systolic gradient of 26 mm Hg (10-60) was reduced to 4 mm Hg (0-20) postprocedure (P < 0.001). There were no deaths on median follow up of 11.5 months (1-56). One patient developed aortic rupture during the procedure and required emergency surgery. CONCLUSION: Covered stent implantation for treatment of aortic coarctation is safe and highly effective in selected patients. Self-expanding stent grafts may be preferable to balloon expandable stents when there is aneurysm formation in the setting of aortopathy. PMID- 18412236 TI - Carotid artery revascularization in high surgical risk patients with the NexStent and the Filterwire EX/EZ: 1-year results in the CABERNET trial. AB - OBJECTIVE: The multicenter, single-arm CABERNET trial evaluated outcomes in high surgical-risk patients with carotid artery stenosis treated with the NexStent plus FilterWire EX/EZ Emboli Protection System. BACKGROUND: For patients at high surgical risk, carotid artery stenting (CAS) offers a less invasive alternative to carotid endarterectomy (CEA). METHODS: The trial enrolled 454 high-surgical risk patients with carotid stenosis by angiography > or = 50% for symptomatic patients and > or = 60% for asymptomatic patients. The comparator primary endpoint was the 1-year major adverse event (MAE, defined as any death, stroke, or myocardial infarction [MI]) rate. It was compared with a proportionally weighted objective performance criterion (OPC) of 12.1% representative of published CEA results in similar patients plus a prespecified noninferiority margin (delta) of 4%. A second primary endpoint was the composite rate of 30-day MAE plus late (31-365 days) ipsilateral stroke. RESULTS: Symptoms of carotid stenosis were present in 24.2% of patients; 36.6% of patients were considered high-surgical-risk due to comorbid risk factors and 63.4% due to anatomic risk factors. The rate of 30-day MAE plus late ipsilateral stroke was 4.7% (20/438). The comparator primary endpoint of 1-year MAE was 11.6% (51/438) and was noninferior to the OPC of 12.1% (95% upper confidence interval of 14.5% versus OPC plus delta of 16.1%, P = 0.005). Late ipsilateral stroke was 0.7% and target vessel revascularization at 1 year was 2.4%. CONCLUSIONS: The CABERNET trial demonstrates that CAS with NexStent and FilterWire is noninferior to (equivalent or better than) traditional CEA at 1 year in high-surgical-risk patients based on historical controls. PMID- 18412237 TI - Thrombectomy during PCI for acute myocardial infarction: are the randomized controlled trial data relevant to the patients who really need this technique? AB - Macro and microembolization during percutaneous coronary intervention (PCI) in ST elevation acute myocardial infarction (STEAMI) is frequent and may result in obstruction of the microvascular network with subsequent reduction in efficacy of reperfusion. Numerous mechanistic studies have shown that the presence and size of the culprit thrombus is the most powerful predictor of incidence of embolization and slow flow/no reflow. Techniques that have been used to reduce the incidence of these events include thrombectomy devices and embolic protection devices. Although numerous prospective randomized clinical trials have been performed to evaluate the role of thrombectomy devices in patients with STEAMI, the results of these trials are conflicting and they speak to both sides of the controversy. The Achilles heal of the majority of these trials is the premise that thrombectomy devices should be routinely used in all patients presenting with STEAMI even irrespective of the presence and size of the thrombus. Such a hypothesis is naively optimistic and it ignores the basic knowledge available to us regarding the relationship between thrombus burden and embolization. Nonetheless, clinicians are faced every day with the reality of making difficult decisions on how to best treat patients presenting with STEAMI and large thrombus burden. The current available "evidence-based medicine" cannot and should not be generalized to these patients because only a minority of these patients was included in these randomized clinical trials. In these patients, thrombectomy devices should be strongly considered as an integral part of the armamentarium available to reduce thrombus burden prior to definitive treatment. Whether a future clinical trial will provide a definitive answer in terms of clinical outcome difference is doubtful because such a trial will need to include large number of selected patients with STEAMI who both have large amount of myocardium at jeopardy and large thrombus burden, a difficult and possibly undoable study. PMID- 18412239 TI - Late outcomes of drug-eluting versus bare metal stents in saphenous vein grafts: Propensity score analysis. AB - OBJECTIVE: To compare late outcomes with the routine use of drug-eluting stents (DES) compared with bare-metal stents (BMS) during percutaneous intervention (PCI) of saphenous vein grafts (SVGs). BACKGROUND: Safety concerns >1 year from stent implantation have been raised about DES used for PCI of SVGs in a small randomized clinical trial. However, there are few studies comparing late outcomes of DES and BMS in routine clinical practice. METHODS: Clinical outcomes (nonfatal MI, cardiac mortality) were assessed in 74 consecutive patients who received BMS and 74 consecutive propensity score matched patients that received DES for PCI of SVGs. Clinical follow-up was censored at 2 years +/- 30 days for both stent groups. RESULTS: At 2 years, the hazard ratio for DES compared with BMS for PCI of SVGs for target vessel revascularization was 0.54 (0.21-1.36), nonfatal MI or cardiac death was 0.68 (0.27-1.68), cardiac mortality 1.19 (0.32-4.45), and stent thrombosis 0.49 (0.09-2.66). Similar outcomes were observed stratified by propensity score quintile. CONCLUSIONS: The routine clinical use of DES for PCI of SVGs was associated with a safety profile that was similar to that of bare metal stents with a clear trend toward a less frequent need for reinterventions. PMID- 18412238 TI - Long-term clinical outcomes of real-world experience using sirolimus-eluting stents in saphenous vein graft disease. AB - OBJECTIVE: To evaluate the long-term clinical outcomes of patients undergoing percutaneous coronary intervention for saphenous vein graft (SVG) disease. Specifically, we compared clinical endpoints of patients who received sirolimus eluting stents (SES) versus bare-metal stents (BMS) for SVG disease. BACKGROUND: A recent small randomized-controlled trial (RCT) reported increased mortality with the use of SES in SVG disease. METHODS: We retrospectively identified patients who underwent SES placement for a SVG lesion(s) at our institutions over a 4-year period. The procedural and medical records were reviewed to identify predetermined clinical outcomes. RESULTS: 318 patients who underwent SES placement for a SVG lesion were identified. 7 patients were lost to follow-up. 141/311 patients (45%) received SES, while 170/311 (55%) received BMS. At a mean follow-up of 34 months, there was a reduction in target lesion revascularization (TLR) (7% vs. 14%, P = 0.07) without an increased risk of mortality (6% vs. 12%, P = 0.06) in patients who received SES compared to patients who received BMS. When compared to the recent RCT's SES patients at long-term follow-up, our SES patients had significantly less mortality; rates of myocardial infarction, TLR, target vessel revascularization, and major adverse cardiac events; and were more likely to be taking dual antiplatelet and statin medications. CONCLUSION: Our results support that SES used in SVG lesions result in a reduction in TLR without an increased risk of mortality, and therefore may be an equally safe and feasible technique for revascularization with excellent long-term clinical outcomes. These patients may benefit from prolonged dual antiplatelet and statin medication regimens. PMID- 18412240 TI - Pleurocutaneous fistula after tube thoracostomy: sonographic findings. AB - Pleurocutaneous fistula is defined as a pathologic communication between the pleural space and the subcutaneous tissues. It can occur as a complication of an infectious process, neoplasm, foreign body aspiration, or iatrogenic procedures. Diagnosis is usually made on imaging studies, such as CT. We describe a 72-year old woman with esophageal cancer who underwent chest tube placement for postesophagectomy drainage. After removal of the chest tube, a right chest wall mass was noted. Transcutaneous sonographic evaluation confirmed the diagnosis of pleurocutaneous fistula. PMID- 18412241 TI - Dietary prevention of malignant glioma aggressiveness, implications in oxidant stress and apoptosis. AB - Our study explored the influence of diet on gliomagenesis and associated systemic effects (SE) in rats. The experimental diet contained various ingredients supposed to interfere with carcinogenesis, mainly phytochemicals (PtcD for phytochemical diet) and its effects were compared to those of the same diet without the phytochemicals (BD for basal diet). Glioma was induced by ethylnitrosourea to pregnant females fed the diets from the start of gestation until the moment of sacrifice of the offpsrings. In male rats fed the PtcD or the BD the incidence of gliomas was markedly reduced compared to rats fed a standard diet (StD). In females this effect was weaker and was limited to the PtcD. A significant proportion of rats with brain tumors and fed the StD exhibited SE evidenced by weight loss, a shorter survival, reduction in liver weight and an increased proportion of liver mitochondria, effects that were not observed in their counterpart fed PtcD. Comparison of the expression of genes involved in the balance proliferation/apoptosis and in the response to oxidative stress in male brain tumors showed that the prevention of SE was associated with an increase in bcl-2 and catalase and a decrease in ki-67, sod-1 and sod-2 transcripts. These results show that the degree of agressiveness of gliomas can be modulated by dietary interventions and suggest that some phytochemicals with antioxidant properties could participate to the mechanism. PMID- 18412242 TI - Fetal growth and the risk of childhood CNS tumors and lymphomas in Western Australia. AB - The etiology of childhood cancers is largely unknown, although the early age at diagnosis has led to particular interest in in utero and perinatal factors. Birth weight is the most frequently studied perinatal factor in relation to risk of childhood cancers, and results have been inconsistent. We investigated whether the risk of CNS tumors and lymphomas in children was associated with three measures of the appropriateness of intra-uterine growth: proportion of optimal birth weight (POBW), birth length (POBL) and weight for length (POWFL). A cohort of 576,633 infants born in Western Australia in 1980-2004 were followed from birth to diagnosis of a CNS tumor (n = 183) or lymphoma (n = 84) before age 15, death, or December 31, 2005, and analyzed with Cox regression. Overall, there was little evidence of any association between fetal growth and risk of CNS tumors, although risk of ependymoma/choroid plexus tumors was positively associated with POBL and negatively associated with POWFL. The risk of Hodgkin and Burkitt lymphoma increased with increasing fetal growth among boys only, whereas the increased risk observed with non-Hodgkin lymphoma was only in girls. These associations between fetal growth and disease risk were also observed among children not classified as high birth weight, suggesting that accelerated growth is more important than birth weight per se. Results were similar when cases were compared with their unaffected siblings, suggesting that the increased growth associated with cancer risk was not general to the family. The associations we observed are consistent with causal pathways involving fetal growth factors. PMID- 18412243 TI - TNF-alpha-inducing protein, a carcinogenic factor secreted from H. pylori, enters gastric cancer cells. AB - TNF-alpha inducing protein (Tip alpha) is secreted from Helicobacter pylori (H. pylori): it is a potent inducer of TNF-alpha and chemokine genes, mediated through NF-kappaB activation, and it also induces tumor-promoting activity in Bhas 42 cells. To investigate the carcinogenic mechanisms of H. pylori with Tip alpha, we first examined how Tip alpha acts on gastric epithelial cells. We found that fluorescent-Tip alpha specifically bound to, and then entered, the cells in a dose- and temperature-dependent manner, whereas deletion mutant of Tip alpha (del-Tip alpha), an inactive form, neither bound to nor entered the cells, suggesting the presence of a specific binding molecule. Mutagenesis analysis of Tip alpha revealed that a dimer formation of Tip alpha with a disulfide bond is required for both specific binding and induction of TNF-alpha gene expression. A confocal laser scanning microscope revealed some Tip alpha in the nuclei, but del Tip alpha was not present, which indicated that an active form of Tip alpha can penetrate the nucleus and may be involved in the induction of TNF-alpha gene expression. Examination of Tip alpha production and secretion in 28 clinical isolates revealed that H. pylori obtained from gastric cancer patients secreted Tip alpha in significantly higher amounts than did H. pylori from patients with chronic gastritis, suggesting that Tip alpha is an essential factor in H. pylori inflammation and cancer microenvironment in the human stomach. Tip alpha is thus a new carcinogenic factor of H. pylori that can enter the nucleus through a specific binding molecule, and its mechanism of action is completely different from that of CagA. PMID- 18412244 TI - Degradation of HPV20E6 by p53: Delta Np63 alpha and mutant p53R248W protect the wild type p53 mediated caspase-degradation. AB - The E6 and E7 proteins of human papillomaviruses (HPV) play a crucial role in the pathogenesis of malignant tumors. E6 protein of high-risk mucosal papillomaviruses targets a number of proteins for proteosomal degradation through complex formation with ubiquitin ligase E6AP. These proteins include, amongst others, p53, paxillin and PDZ-domain proteins e.g. Dlg and MAGUK. The mechanism by which the E6 protein of cutaneous HPV types interacts with cellular proteins to induce either benign or malignant cutaneous lesions, has not been elucidated, although extensive ultraviolet exposure and mutated p53 (hot-spot mutations) are known to be associated with non-melanoma skin cancer. We demonstrate two mechanisms in which HPV20E6 may be involved in the infected cell. One pathway is the wtp53 mediated degradation of HPV20E6 through caspase-3. Mutated p53R248W and Delta Np63 alpha, as well as other unknown proteins involved in proteosome dependent degradation, convey a protective effect on HPV20E6 under these conditions. This unveils a remarkable opposite regulation to the well-known mechanism of E6-E6AP mediated degradation of p53 for mucosal HPV types. In a second interaction, ectopically expressed HPV20E6 induces cleavage of procaspase 3 to active caspase-3. We demonstrate, in addition, in vivo binding of HPV20E6 to the intermediate filament vimentin. PMID- 18412245 TI - Cancer and mortality among users and nonusers of snus. AB - Scandinavian moist snuff (snus) is claimed to be a safer alternative to smoking. We aimed to quantify cancer incidence among male snus users and to shed light on the net health outcome by studying their overall mortality. A cohort, comprised of 9,976 men who participated in a population-based survey, was compiled in 1973 74. Follow-up until January 31, 2002, was accomplished through record-linkages with nation-wide and essentially complete registers of demographics, cancer and causes of deaths. Adjusted relative risks among exposed relative to unexposed men were estimated using Cox proportional hazards regression. The cohort members contributed more than 220,000 person-years at risk for cancer. A statistically significant increase in the incidence of the combined category of oral and pharyngeal cancer among daily users of snus (incidence rate ratio 3.1, 95% confidence interval 1.5-6.6) was found. Overall mortality was also slightly increased (hazard ratio 1.10, 95% confidence interval 1.01-1.21). Although the combined previous literature on snus and oral cancer weigh toward no association, this population-based prospective study provided suggestive evidence of snus related risks that cannot be lightly ignored. PMID- 18412246 TI - Opposing effects of fibrosarcoma cell-derived IL-1 alpha and IL-1 beta on immune response induction. AB - There is evidence that cell-associated IL-1 alpha supports immune response induction. Here we explored the impact of malignant cell-derived IL-1 on immunogenicity, immune response induction and tumor-induced immunosuppression using 3-methylcholanthrene-induced fibrosarcoma lines derived from wild-type (wt), IL-1 alpha-, IL-1 beta- or IL-1a beta-knockout (IL-1 alpha(-/-), IL-1 beta( /-), IL-1 alphabeta(-/-)) C57BL6 mice. The wt, IL-1 alpha(-/-), IL-1 beta(-/-) and IL-1 alphabeta(-/-) fibrosarcoma lines express MHC class I molecules at a high level. The lines do not differ in their susceptibility toward NK cells, macrophages, and allogeneic CTL, or in their capacity as stimulators of an allogeneic response. However, IL-1 beta(-/-) tumors rarely grow in the syngeneic host, which is the consequence of a strong T helper and CTL response induction by IL-1 alpha-competent, IL-1 beta(-/-) tumors. On the other hand, IL-1 beta competent, IL-1 alpha(-/-) tumors strongly assist CD11b(+)Gr-1(+) myeloid-derived suppressor cell and regulatory T cell expansion, which both suppress with high efficacy activated T helper cell proliferation and CTL lysis. In IL-1 alphabeta( /-) tumors, the absence of IL-1 alpha becomes decisive, i.e. despite reduced suppressor cell recruitment, tumor growth was unimpaired due to inefficient immune response induction. Thus, sarcoma cell-derived IL-1 alpha and IL-1 beta do not act in concert. Induction of a strong immune response by IL-1 alpha demands therapeutic exploitation, which may become more efficient if systemic induction of immunosuppression by IL-1 beta can also be circumvented. PMID- 18412247 TI - Determinants of overall quality of life in preterminal cancer patients. AB - Recently, a few studies reported that fatigue was a predominant contributor to patient-perceived overall QoL in patients with different types of cancer in a relatively early stage of disease. In the present study, we aimed to investigate whether fatigue is also a major contributor to overall QoL in preterminal cancer patients. Ninety-eight preterminal cancer patients, mainly lung (44%) and gastrointestinal cancer (25%), with an estimated life expectancy of 1-6 months were included. QoL domains as well as overall QoL were measured using the EORTC QLQ-C30 questionnaire. Data were analysed by Pearson's correlations and multivariate modelling. Fatigue showed the strongest correlation with overall QoL (r = -0.63, p < 0.001), followed in decreasing order by role functioning (r = 0.53), physical functioning (r = 0.47), social functioning (r = 0.44), nausea (r = -0.37), cognitive functioning (r = 0.33), appetite loss (r = -0.31), dyspnea (r = -0.26) and emotional functioning (r = 0.24). Multivariate analysis confirmed that the fatigue scale paid by far the highest individual contribution to overall QoL (standardized regression coefficient (SRC): -0.41, p = 0.002), followed by social functioning (SRC: 0.18, p = 0.05). None of the other domains or symptom scales contributed independently to overall QoL. Our results clearly demonstrate that, in preterminal cancer patients, fatigue is a major contributor of overall QoL, corroborating reports in cancer patients in earlier disease stages. PMID- 18412248 TI - Utilization of an aspiration thrombectomy catheter (Pronto) to treat acute atherothrombotic embolization during percutaneous revascularization of the lower extremity. AB - Acute limb ischemia is typically a surgical emergency and can occur secondary to distal embolization during catheter-based percutaneous revascularization. We present a case of acute atherothrombotic embolization to the left lower extremity after stenting of the left proximal popliteal and anterior tibial arteries. This was successfully treated with the use of the coronary aspiration thrombectomy device Pronto (Vascular Solutions, Minneapolis, MN) resulting in flow restoration. PMID- 18412249 TI - Novel guidewire-based stent delivery system: Examination by intravascular ultrasound. AB - This report describes the first-in-man experience with a novel 0.014-in. guidewire-based, self-expanding stent delivery system designed for small or tortuous coronary arteries that may be difficult to access with conventional stent-delivery systems. PMID- 18412250 TI - Avoiding pacemaker lead entrapment during PFO closure. AB - We present a case of a patient with a history of cryptogenic stroke and implanted dual-chamber pacemaker referred for transcatheter closure of patent foramen ovale (PFO). During the procedure ventricular lead of the pacemaker was found to run alongside the PFO. This interrupted correct positioning and deployment of the Amplatzer occluder without sandwiching of the lead. A self-prepared pigtail catheter with a cut off tip was used to move the lead away from the PFO. Subsequently, an occluder was successfully placed. In conclusion, difficulties in Amplatzer occluder placement in patients with implanted pacemaker may be overcome by a manoeuvre which prevents lead entrapment by the device. PMID- 18412251 TI - Feasibility of patent foramen ovale closure with no-device left behind: first-in man percutaneous suture closure. AB - OBJECTIVES: This study evaluated the feasibility of percutaneous patent foramen ovale (PFO) closure using a transcatheter suture (Superstitch), leaving no device behind. BACKGROUND: PFO has been implicated in cryptogenic strokes and migraine with auras. Percutaneous PFO closure, being less invasive than surgical closure, is increasingly performed. There are, however, early and long-term risks including device embolization, fracture, thrombosis, or infection, erosions into the free atrial wall and aorta, arrhythmias, and death. Furthermore, device implantation may complicate future percutaneous access to the left atrium. Partially reabsorbable devices and tissue welding to close PFO have recently been introduced. The first-in-man transcatheter suture closure of a PFO in an 18-year old female with chronic migraine with aura and a well documented stroke is described. METHODS: The right femoral vein was cannulated under mild sedation and local anesthesia. Using intracardiac echocardiography imaging, bubble study demonstrated a right-to-left shunt through the PFO at rest. A Superstitch device was advanced across the PFO and sutures were delivered through the septum primum and secundum. The sutures were exteriorized and a knot was advanced to the right atrial septum and cut. RESULTS: Bubble study confirmed successful PFO suture closure. Transesophageal and transthoracic echocardiograms with bubble studies at 1 and 2 months, respectively showed complete closure with no right-to-left shunt even during Valsalva maneuver. At 6 months, the patient remained free of symptoms or migraine episodes. CONCLUSION: Percutaneous transcatheter suture closure of a PFO can be successfully achieved with no residual shunt and leaving no device behind. Technological refinements are required for wider scale use. PMID- 18412252 TI - Spectrum of gait impairments in presymptomatic and symptomatic Huntington's disease. AB - The purpose of this study was to quantify gait impairments in presymptomatic and symptomatic Huntington's disease (HD) subjects, and examine sensitivity of gait measures. Our sample (n = 65) included presymptomatic mutation carriers (PMC) (n = 15), symptomatic HD subjects (SHD) (n = 30) and healthy controls (n = 20). Participants were requested to walk at their preferred speed on a computerized walkway that recorded spatiotemporal variables. We administered the Unified HD Rating Scale (UHDRS) for PMC and SHD. PMC demonstrated decreased gait velocity (P < 0.01), stride length (P < 0.008), and increased time in double support (P < 0.001); and demonstrated higher variability in stride length (P < 0.01) and step time (P < 0.004) compared with controls. These impairments worsened with increasing disease severity for SHD. Gait impairments were correlated with predicted years to onset in PMC (velocity = -0.65; cadence = -0.70, step time = 0.71) and demonstrated high sensitivity and specificity in distinguishing between controls and mutation carriers. In contrast, UHDRS scores did not reveal impairments in gait and balance. Gait bradykinesia and dynamic balance impairments begin in the presymptomatic stage of HD and continue to worsen in the symptomatic stages. Gait measures are sensitive in differentiating between mutation positive and negative individuals even when impairments were not detected by clinical neurological examination. (c) 2008 Movement Disorder Society. PMID- 18412253 TI - Localization of betav tubulin in the cochlea and cultured cells with a novel monoclonal antibody. AB - Tubulin, the dimeric structural protein of microtubules, is a heterodimer of alpha and beta subunits; both alpha and beta exist as numerous isotypes encoded by different genes. In vertebrates the sequence differences among the beta(I), beta(II), beta(III), beta(IV) and beta(V) isotypes are highly conserved in evolution, implying that the isotypes may have functional significance. Isotype specific monoclonal antibodies have been useful in determining the cellular and sub-cellular distributions and possible functions of the beta(I), beta(II), beta(III), and beta(IV) isotypes; however, little is known about the beta(V) isotype. We here report the creation and purification of a monoclonal antibody (SHM.12G11) specific for beta(V). The antibody was designed to be specific for the C-terminal sequence EEEINE, which is unique to rodent and chicken beta(V). The antibody was found to bind specifically to the C-terminal peptide EEEINE, and does not cross-react with the carboxy-termini of either alpha-tubulin or the other beta-tubulin isotypes. However, the antibody also binds to the peptide EEEVNE, but not to the peptide EEEIDG, corresponding respectively to the C terminal peptides of bovine and human beta(V). Immunofluorescence analysis indicates that beta(V) is found in microtubules of both the interphase network and the mitotic spindle. In gerbils, beta(V) also occurs in the cochlea where it is found largely in the specialized cells that are unique in containing bundled microtubules with 15 protofilaments. PMID- 18412254 TI - Dynamic rearrangement of F-actin organization triggered by host-specific plant signal is linked to morphogenesis of Aphanomyces cochlioides zoospores. AB - Cochliophilin A (5-hydroxy-6,7-methylenedioxyflavone), a root releasing host specific plant signal triggers chemotaxis and subsequent morphological changes in pathogenic Aphanomyces cochlioides zoospores before host penetration. The present study illustrates time-course changing patterns of cytoskeletal filamentous actin (F-actin) organization in the zoospores of A. cochlioides during rapid morphological changes (encystment and germination) after exposure to cochliophilin A. Confocal laser scanning microscopic analysis revealed that F actin microfilaments remained concentrated at ventral groove and diffusely distributed in peripheral cytoplasm of the zoospore. These microfilaments dramatically rearranged and changed into granular F-actin plaques interconnected with fine arrays during encystment. A large patch of actin arrays accumulated at one pole of the cystospores just before germination. Then the actin plaques moved to the emerging germ tube where a distinct cap of microfilaments was seen at the tip of the emerging hypha. Zoospores treated with an inhibitor of F-actin polymerization, latrunculin B or motility halting and regeneration inducing compound nicotinamide, displayed different patterns of F-actin in both zoospores and cystospores than those obtained by the induction of cochliophilin A. Collectively, these results indicate that the host-specific plant signal cochliophilin A triggers a dynamic polymerization/depolymerization of F-actin in pathogenic A. cochlioides zoospores during early events of plant peronosporomycete interactions. PMID- 18412256 TI - Energetics of K+ permeability through Gramicidin A by forward-reverse steered molecular dynamics. AB - The estimation of ion channel permeability poses a considerable challenge for computer simulations because of the significant free energy barriers involved, but also offers valuable molecular information on the ion permeation process not directly available from experiments. In this article we determine the equilibrium free energy barrier for potassium ion permeability in Gramicidin A in an efficient way by atomistic forward-reverse non-equilibrium steered molecular dynamics simulations, opening the way for its use in more complex biochemical systems. Our results indicate that the tent-shaped energetics of translocation of K+ ions in Gramicidin A is dictated by the different polarization responses to the ion of the external bulk water and the less polar environment of the membrane. PMID- 18412255 TI - The incidence of clinically significant contrast-induced nephropathy following non-emergent coronary angiography. AB - OBJECTIVES: The primary aim of this study was to assess the incidence of clinically significant contrast-induced nephropathy (CIN) among patients undergoing non-emergent coronary angiography. BACKGROUND: Although retrospective analyses have emphasized the association of CIN with adverse patient outcomes, the actual incidence of clinically significant CIN following non-emergent coronary angiography is not clear. METHODS: We prospectively identified patients with baseline estimated glomerular filtration rates (eGFR) < 60 ml/min/1.73 m(2) undergoing non-emergent coronary angiography. We measured serum creatinine 48-96 hr following angiography and assessed the incidence of CIN using two definitions, a rise in Scr > or = 25% and > or = 0.5 mg/dl. We tracked the need for dialysis, hospitalization related to kidney injury, and 30-day mortality to examine the association of CIN with these outcomes. RESULTS: We enrolled 181 patients with a median eGFR of 52 ml/min/1.73 m(2). Of the 165 patients (91%) with post-procedure Scr data, the incidence of CIN was 6.1-8.5%. One patient required dialysis (0.55%) and one (0.55%) died within 30 days. Although 38 patients required hospital admission, CIN was not associated with the need for hospitalization. Patients with an increase in Scr > or = 25% demonstrated a trend toward increased risk for 30-day mortality (P = 0.09), whereas those with increments in Scr > or = 0.5 mg/dl had a marginally higher risk for dialysis (P = 0.06) and 30-day mortality (P = 0.06), although these associations failed to meet the level of statistical significance. CONCLUSIONS: Biochemically defined CIN occurs in a small, but notable proportion of patients undergoing non-emergent coronary angiography. However, clinically significant CIN is very uncommon. PMID- 18412257 TI - A QM/MM study of proton transport pathways in a [NiFe] hydrogenase. AB - A theoretical QM/MM study of the [NiFe] hydrogenase from Desulfovibrio fructosovorans has been performed to investigate possible routes of proton transfer between the active site and the protein surface. We obtained the minimum energy paths, with a modified version of the nudged elastic band method, for a set of proposed pathways. The calculations were carried out for the crystallographic structure and for several structures of the protein obtained from a molecular dynamics simulation. The results show one of the studied pathways to be preferred for transport from the active site to the surface, but the preference is not so strong when transport occurs in the opposite direction. PMID- 18412258 TI - Discriminative learning for protein conformation sampling. AB - Protein structure prediction without using templates (i.e., ab initio folding) is one of the most challenging problems in structural biology. In particular, conformation sampling poses as a major bottleneck of ab initio folding. This article presents CRFSampler, an extensible protein conformation sampler, built on a probabilistic graphical model Conditional Random Fields (CRFs). Using a discriminative learning method, CRFSampler can automatically learn more than ten thousand parameters quantifying the relationship among primary sequence, secondary structure, and (pseudo) backbone angles. Using only compactness and self-avoiding constraints, CRFSampler can efficiently generate protein-like conformations from primary sequence and predicted secondary structure. CRFSampler is also very flexible in that a variety of model topologies and feature sets can be defined to model the sequence-structure relationship without worrying about parameter estimation. Our experimental results demonstrate that using a simple set of features, CRFSampler can generate decoys with much higher quality than the most recent HMM model. PMID- 18412259 TI - Prostaglandin E2 modulates the expression of antimicrobial peptides in the fat body and midgut of Anopheles albimanus. AB - Prostaglandins (PGs) participate in the regulation of vertebrate and in at least six insect orders' immune responses. We identified PGE2 in midgut, fat body, Malpighian tubules, and ovarioles of Anopheles albimanus (Aa) mosquitoes. Our data indicate that PGE2 synthesis in cultured midguts responds to the presence of two bacterial species, Micrococcus luteus and Klebsiella pneumoniae. The production of mRNA coding for antimicrobial peptides Aa-Attacin, Aa-Cecropin, and Aa-Gambicin was observed in cultured fat bodies and midguts. The production of these messengers was reduced in the presence of dexamethasone, and this effect was reversed by arachidonic acid. Adding PGE2 to cultures resulted in increased Aa-cecropin mRNA and decreased Aa-attacin and Aa-gambicin mRNAs. PMID- 18412260 TI - Low recurrence of preexisting extrahepatic malignancies after liver transplantation. AB - The incidence of de novo malignancies is increased in organ transplant recipients, and patients with hepatic carcinomas are at high risk for tumor recurrence after liver transplantation. Data about recurrent cancer after orthotopic liver transplantation (OLT) in patients with a history of nonhepatic malignancy are very limited. We retrospectively analyzed data from 606 adult OLT recipients and identified 37 patients (6.1%) with a preexisting extrahepatic malignancy. In the same group, 43 patients (7.0%) developed de novo cancer. Preexisting malignancies included 26 solid tumors and 11 hematological malignancies, including 7 patients with Budd-Chiari syndrome due to myeloproliferative disorders (MPDs). Patients had been selected for OLT because of the expected good prognosis of their preexisting malignancy. Except for 3 patients, recipients were tumor-free at OLT. The median interval from tumor diagnosis to OLT was 44 months (range, <1-321). After a median follow-up of 66 months post transplantation (range, 4-131), all but 1 recipient with incidental colon carcinoma were free of recurrence. No patient with MPD showed leukemic transformation, whereas a patient with neurofibromatosis experienced growth of skin fibromas. Our data and an included review of published OLT recipients with preexisting malignancies have enabled us to show that recurrence rates are comparable for nontransplanted patients and renal-transplant recipients. In conclusion, cancer recurrence is low if OLT recipients are carefully selected. Therefore, previous extrahepatic malignancy should not be considered a contraindication for OLT per se, but the oncologic/hematologic prognosis should be considered, particularly with respect to the current 5-year survival rate of OLT. PMID- 18412261 TI - Coarse-grained dynamics of the receiver domain of NtrC: fluctuations, correlations and implications for allosteric cooperativity. AB - Receiver domains are key molecular switches in bacterial signaling. Structural studies have shown that the receiver domain of the nitrogen regulatory protein C (NtrC) exists in a conformational equilibrium encompassing both inactive and active states, with phosphorylation of Asp54 allosterically shifting the equilibrium towards the active state. To analyze dynamical fluctuations and correlations in NtrC as it undergoes activation, we have applied a coarse-grained dynamics algorithm using elastic network models. Normal mode analysis reveals possible dynamical pathways for the transition of NtrC from the inactive state to the active state. The diagonalized correlation between the inactive and the active (phosphorylated) state shows that most correlated motions occur around the active site of Asp54 and in the region Thr82 to Tyr101. This indicates a coupled correlation of dynamics in the "Thr82-Tyr101" motion. With phosphorylation inducing significant flexibility changes around the active site and alpha3 and alpha4 helices, we find that this activation makes the active-site region and the loops of alpha3/beta4 and alpha4/beta5 more stable. This means that phosphorylation entropically favors the receiver domain in its active state, and the induced conformational changes occur in an allosteric manner. Analyses of the local flexibility and long-range correlated motion also suggest a dynamics criterion for determining the allosteric cooperativity of NtrC, and may be applicable to other proteins. PMID- 18412262 TI - Structure of YraM, a protein essential for growth of Haemophilus influenzae. AB - Nontypeable Haemophilus influenzae is an obligate human parasite that often causes middle ear infections in children and exacerbates chronic obstructive pulmonary disorder, the fourth leading cause of death in the United States. There are no effective vaccines available for this strain. The lipoprotein YraM (gene HI1655) was identified as essential for the growth and viability of H. influenzae but its function is unknown. Sequence comparisons showed that YraM is a fusion of two protein modules. We grew crystals of the carboxyl-terminal module of YraM comprising residues 257-573 (YraM-C), phased the diffraction data by the multiwavelength anomalous diffraction technique, and refined the model to a crystallographic R-factor of 0.16 (R(free) = 0.19) with data to 1.35 A resolution. The two-domain structure of YraM-C adopts a fold similar to that observed for the open, unliganded forms of several periplasmic binding proteins (PBPs) involved in bacterial active transport. Sequence alignments of YraM homologues from other Gram-negative species showed that the most conserved residues of YraM-C cluster between the two domains in the location where other PBPs bind their cognate ligand. Modeling of YraM-C into a closed conformation similar to the leucine-bound form of the Leu/Ile/Val-binding protein (LIVBP) shows a putative binding pocket larger than the leucine-binding site in LIVBP. The pocket has both polar and nonpolar surfaces, with the latter located in the same area where a leucine side chain binds to LIVBP. We discuss possible biological functions of YraM considering its predicted location in the outer membrane, a novel place for such a binding protein. PMID- 18412263 TI - Crystal structure of the methyltransferase domain of human TARBP1. PMID- 18412264 TI - Functional imaging: I. Relative predictive value of intracranial electroencephalography. AB - OBJECTIVE: To gain information on the predictive and prognostic value of magnetic source imaging (MSI), 2-[18F]fluoro-2-deoxy-D-glucose positron emission tomography ((18)FDG-PET), and ictal single-photon emission computed tomography (SPECT) as compared with intracranial electroencephalography (ICEEG) localization in epilepsy surgery. METHODS: This work was part of a cohort study of epilepsy surgery candidates not sufficiently localized with noninvasive studies. Of 160 patients enrolled over 4 years, 77 completed ICEEG seizure monitoring. Sensitivity, specificity, and predictive values relative to ICEEG were computed for each modality. RESULTS: Seizures were not captured in five patients. Of the 72 diagnostic ICEEG studies, seizure localization results were 74% localized, 10% multifocal, and 17% nonlocalized. Sixty-one percent were localized to neocortical regions. Depending on patient subgroup pairs, sensitivity ranged from 58 to 64% (MSI), 22 to 40% (PET), and 39 to 48% (SPECT); specificity ranges were 79 to 88% (MSI), 53 to 63% (PET), and 44 to 50% (SPECT). Gains in diagnostic yield were seen only with the combination of MSI and PET or MSI and ictal SPECT. Localization concordance with ICEEG was greatest with MSI, but a significant difference was demonstrated only between MSI and PET. Moderate redundancy was seen between PET and ictal SPECT (kappa = 0.452; p = 0.011). INTERPRETATION: Conclusively positive MSI has a high predictive value for seizures localized with ICEEG. Diagnostic gain may be achieved with addition of either PET or ictal SPECT to MSI. Diagnostic values for imaging tests are lower than "true values" because of the limitations of ICEEG as a gold standard. PMID- 18412265 TI - Analysis of Lrrk2 R1628P as a risk factor for Parkinson's disease. AB - Common genetic variants that increase the risk for Parkinson's disease may differentiate patient subgroups and influence future individualized therapeutic strategies. Herein we show evidence for leucine-rich repeat kinase 2 (LRRK2) c.4883G>C (R1628P) as a risk factor in ethnic Chinese populations. A study of 1,986 individuals from 3 independent centers in Taiwan and Singapore demonstrates that Lrrk2 R1628P increases risk for Parkinson's disease (odds ratio, 1.84; 95% confidence interval, 1.20-2.83; p = 0.006). Haplotype analysis suggests an ancestral founder for carriers approximately 2,500 years ago. These findings support the importance of LRRK2 variants in sporadic Parkinson's disease. Ann Neurol 2008. PMID- 18412266 TI - ANGIOCARE: an automated system for fast three-dimensional coronary reconstruction by integrating angiographic and intracoronary ultrasound data. AB - OBJECTIVES: The development of an automated, user-friendly system (ANGIOCARE), for rapid three-dimensional (3D) coronary reconstruction, integrating angiographic and, intracoronary ultrasound (ICUS) data. METHODS: Biplane angiographic and ICUS sequence images are imported into the system where a prevalidated method is used for coronary reconstruction. This incorporates extraction of the catheter path from two end-diastolic X-ray images and detection of regions of interest (lumen, outer vessel wall) in the ICUS sequence by an automated border detection algorithm. The detected borders are placed perpendicular to the catheter path and established algorithms used to estimate their absolute orientation. The resulting 3D object is imported into an advanced visualization module with which the operator can interact, examine plaque distribution (depicted as a color coded map) and assess plaque burden by virtual endoscopy. RESULTS: Data from 19 patients (27 vessels) undergoing biplane angiography and ICUS were examined. The reconstructed vessels were 21.3-80.2 mm long. The mean difference was 0.9 +/- 2.9% between the plaque volumes measured using linear 3D ICUS analysis and the volumes, estimated by taking into account the curvature of the vessel. The time required to reconstruct a luminal narrowing of 25 mm was approximately 10 min. CONCLUSION: The ANGIOCARE system provides rapid coronary reconstruction allowing the operator accurately to estimate the length of the lesion and determine plaque distribution and volume. PMID- 18412268 TI - Transcatheter occlusion of pulmonary arteriovenous malformations using the Amplatzer Vascular Plug II. AB - Pulmonary arteriovenous malformations (PAVMs) are a relatively uncommon but potentially life-threatening condition manifested by cyanosis, paradoxical embolization, brain abscess, and rupture. A variety of transcatheter closure devices have been used to occlude PAVMs however risks of device embolization, incomplete closure, or large delivery systems have made each of these methods sub optimal. The Amplatzer Vascular Plug II (AVP II) is a new multisegmented, woven nitinol cylinder that can be deployed through a small delivery catheter. The AVP II differs from the original AVP in having a finer, more tightly woven nitinol frame and three, rather than one occlusive segment. The authors report the first use of the AVP II for occlusion of bilateral large arteriovenous malformations in a patient with hereditary hemorrhagic telangiectasia. PMID- 18412267 TI - Alzheimer and frontotemporal pathology in subsets of primary progressive aphasia. AB - OBJECTIVE: To identify predictors of Alzheimer's disease (AD) versus frontotemporal lobar degeneration pathology in primary progressive aphasia (PPA), and determine whether the AD pathology is atypically distributed to fit the aphasic phenotype. METHODS: Neuropsychological and neuropathological analyses of 23 consecutive PPA autopsies. All had qualitative determination of neurofibrillary tangle (NFT) density. Additional quantitation was done in four of the PPA/AD cases and four AD cases with the typical amnestic dementia of the Alzheimer type. RESULTS: The sample contained mostly logopenic, agrammatic, and mixed forms of PPA. All six agrammatics had frontotemporal lobar degeneration (five of six with tauopathy). Seven of the 11 logopenics had AD. In logopenics, lower memory scores increased the probability of AD, but there were exceptions. The PPA/AD group showed predominance of entorhinal NFT typical of the amnestic dementia of the Alzheimer type. In the small subgroup examined quantitatively, neocortical NFTs were more numerous in the left hemisphere of PPA/AD. However, the asymmetry was low and inconsistent. Neuritic plaques did not display consistent asymmetry. Apolipoprotein E4, a major risk factor for typical AD, did not predict AD pathology in PPA. INTERPRETATION: Subtyping PPA helps to predict AD versus frontotemporal lobar degeneration pathology at the group level. However, our results and the literature also indicate that no clinical predictor is completely reliable in individual patients. The inconsistent concordance of NFT distribution with the asymmetric atrophy and the nonamnestic phenotype also raises the possibility that the AD markers encountered at autopsy in PPA may not always reflect the nature of the initiating neurodegenerative process. PMID- 18412269 TI - Percutaneous transcatheter communication between the pulmonary artery and atrium following an extra-cardiac Fontan: an alternative approach to fenestration avoiding conduit perforation. AB - In a nonfenestrated modified Fontan, transcatheter creation of a fenestration presents technical difficulties, especially with the extra-cardiac modification where a Gore-Tex tube is placed between the inferior caval vein and the pulmonary arteries. The authors describe an alternative approach to perforating the Gore Tex tube to create a fenestration by making a communication between the pulmonary artery and the atrial chamber, thus bridging the two circulations through native tissue. PMID- 18412270 TI - Prospective randomized comparison of sirolimus- versus paclitaxel-eluting stents for the treatment of acute ST-elevation myocardial infarction: pROSIT trial. AB - OBJECTIVE: The aim of this study was to compare effectiveness of the Sirolimus- (SES) and Paclitaxel-eluting stent (PES) in primary angioplasty for acute ST elevation myocardial infarction (STEMI). BACKGROUND: It has been reported that SES and PES have been more effective than bare-metal stents in reducing restenosis and cardiac events in a broad range of patients with coronary artery disease. However, it is unknown whether there may be differences between these two drug-eluting stents in terms of efficacy in the setting of acute STEMI. METHODS: Acute STEMI patients (n = 308) undergoing primary angioplasty were randomly assigned to SES (n = 154) or PES (n = 154) deployment. The routine angiographic follow-up was performed at 6 months and clinical follow-up data was obtained at 12 months. The primary end point was major adverse cardiac events (MACE) including death, reinfarction, stent thrombosis, and target lesion revascularization (TLR) at 12 months. RESULTS: The baseline clinical, angiographic, and procedural characteristics were similar between the 2 groups. Two patients (all from the PES group) experienced stent thrombosis (1 acute and 1 subacute). The SES group revealed lower in-segment restenosis (5.9% vs. 14.8%, P = 0.03) and in-segment late loss (0.09 +/- 0.45 vs. 0.33 +/- 0.68 mm, P = 0.002) than PES group on follow-up angiography. Twelve-month TLR rates (2.6% vs. 6.5%, P = 0.17) were similar between two groups. MACE rates were lower in the SES group than in the PES group, but it did not reach statistical significance (5.8% vs. 11.7%, P = 0.07). CONCLUSION: In the setting of primary angioplasty for STEMI, there were no statistically significant differences between the SES and the PES in terms of 12-month MACE. However, binary angiographic in-segment restenosis and in-segment late loss were significantly lower in the SES group. PMID- 18412271 TI - The COURAGE trial in perspective. AB - The indications for percutaneous coronary intervention (PCI) continue to evolve because of the steady improvement in technology, broadened patient and lesion selection criteria, and new evidence from clinical trials. Recently, the role of PCI in patients with chronic stable angina has received considerable scrutiny and has been the subject of great controversy. In these patients, the goals of therapy include the relief of symptom, treatment of ischemia, and reducing the need for subsequent interventions. Medical therapy is the cornerstone in the management of coronary artery disease and should be optimized in all patients. The COURAGE trial investigated the efficacy of combined PCI and optimal medical therapy (OMT) versus OMT alone in patients with stable disease. The trial confirmed several issues that have been already well delineated: (1) in low risk patients, the hard endpoints of death and MI are relatively infrequent and are not reduced by PCI - for prevention of these, OMT may be sufficient, (2) crossover from OMT to PCI is frequent, even in low risk patients, (3) PCI is very effective in reducing symptoms and myocardial ischemia, and (4) significant untreated ischemia is associated with greater likelihood of death and MI. PMID- 18412272 TI - Drug eluting stents for below the knee lesions in patients with critical limb ischemia : long-term follow-up. AB - OBJECTIVES: The purpose of this study was to assess the long-term limb preservation and/or healing of ulcers in patients with critical limb ischemia (CLI) and severe infrapopliteal atherosclerotic disease treated with drug eluting stents (DES). BACKGROUND: Percutaneous revascularization has become an effective treatment for CLI in patients with infrapopliteal atherosclerotic disease. Recent reports using DES in patients with CLI have documented excellent short-term infrapopliteal vessel patency. Higher primary patency rates in infrapopliteal vessels treated with DES could translate into better long-term clinical outcomes and improved limb salvage rates. METHODS: Twenty-four consecutive patients with CLI (defined as rest pain, nonhealing ulcers, or gangrene) because of severe infrapopliteal disease were treated with DES from August 2004 to June 2006. RESULTS: Procedural success was achieved in 96% (27/28) of targeted lesions. There were no procedure-related deaths, acute vessel thrombosis events, or need for urgent surgical intervention. There was one case of distal embolization. Clinical follow up, ranging 8-34 months, is available for 100% of patients of which 83% (20/24) achieved limb preservation and healing. Angiographic and/or sonographic follow up, ranging 6-34 months, is available in 79% (19/24) of patients of which 95% (18/19) had patent target vessels. CONCLUSIONS: DES is a safe and effective long-term option for CLI due to severe infrapopliteal arterial disease. Long-term vascular patency led to a high rate of limb preservation and low amputation rate. A multicenter trial should further elucidate the role of DES in the treatment of CLI. PMID- 18412275 TI - Intensive cardiac management in patients with trisomy 13 or trisomy 18. AB - Intensive cardiac management such as pharmacological intervention for ductal patency (indomethacin and/or mefenamic acid for closure and prostaglandin E1 for maintenance) and palliative or corrective surgery is a standard treatment for congenital heart defects. However, whether it would be a treatment option for children with trisomy 13 or trisomy 18 syndrome is controversial because the efficacy on survival in patients with these trisomies has not been evaluated. We retrospectively reviewed 31 consecutive neonates with trisomy 13 or trisomy 18 admitted to our neonatal ward within 6 hr of birth between 2000 and 2005. The institutional management policies differed during three distinct periods. In the first period, both pharmacological ductal intervention and cardiac surgery were withheld. In the second, pharmacological ductal intervention was offered as an option, but cardiac surgery was withheld. Both strategies were available during the third period. The median survival times of 13, 9, and 9 neonates from the first, second, and third periods were 7, 24, and 243 days, respectively. Univariate and multivariate analyses confirmed that the patients in the third period survived significantly longer than the others. Intensive cardiac management consisting of pharmacological intervention for ductal patency and cardiac surgery was demonstrated to improve survival in patients with trisomy 13 or trisomy 18 in this series. Therefore, we suggest that this approach is a treatment option for cardiac lesions associated with these trisomies. These data are helpful for clinicians and families to consider in the optimal treatment of patients with these trisomies. PMID- 18412274 TI - Live birth prevalence of Down syndrome in Tottori, Japan, 1980-1999. AB - One hundred sixty-four patients with Down syndrome (DS) were confirmed in Tottori Prefecture, Japan, from 1980 to 1999. The sex ratio of 1.52 (99 males and 65 females) was comparable to that reported in previous studies. The live birth prevalence per 1,000 was 1.52 (95% CI: 1.29-1.75) from 1980 to 1999, with a prevalence of 1.34 (95% CI: 1.05-1.63) recorded between 1980 and 1989, and 1.74 (95% CI: 1.37-2.11) between 1990 and 1999. There was no statistically significant change between these two decades (chi(2)-test). Live birth prevalence in these two decades showed a significant increase (chi(2)-test, P < 0.005) compared with that recorded in 1969-1978 in Tottori Prefecture (0.803, 95% CI: 0.677-0.929). Mean ages of mothers at the birth of a DS patient were 31.0 years in 1980-1989 and 32.4 years in 1990-1999 (t-test, no significant difference). Dispersion analysis on the mean age of mothers at birth for patients born between 1969-1978, 1980-1989, and 1990-1999 showed a significant difference (t-test, P < 0.005), while comparing the mean age of mothers in 1969-1978 to those in 1990-1999 also revealed a significant difference (t-test, P < 0.001). Live birth prevalence has increased due to the rise in fertility rates among older women, although maternal age-specific risk rates remain unchanged. The widespread introduction of induced abortion following prenatal diagnosis decreased live birth prevalence of DS largely in European (and a few Asian) countries after 1990, or kept prevalence steady, despite increasing fertility rates among women aged 30 and over. In contrast, all published studies have reported an increase in live birth prevalence of this syndrome in Japan, probably resulting from the fact that prenatal diagnoses are used only exceptionally in this country (due to the negative attitude toward selection of life in Japanese culture). PMID- 18412276 TI - Predicted prevalence of Down syndrome live births in Japan, 1970-2006. PMID- 18412277 TI - Absence of decussation of the superior cerebellar peduncles in patients with Joubert syndrome. AB - Joubert syndrome (JS) is characterized by absence of decussation of both corticospinal tracts and superior cerebellar peduncles (SCP). Our hypothesis was that evidence for absence of SCP decussation may be found on routine brain MRI studies. Midsagittal T1-weighted images from 20 JS patients and 30 age-matched controls were retrospectively reviewed. An ill-defined area of lower T1 signal in the inferior midbrain was considered the sign of SCP decussation, and its presence or absence was noted. Fractional anisotropy (FA) maps were obtained in three JS patients and four controls. The SCP decussation was not visualized in any subject under 30 months of age. In subjects above 30 months of age the sign of the SCP decussation was absent in all 6 JS patients and present in all 16 controls. FA maps confirmed the absence of the SCP decussation in the JS patients. The SCP decussation is well seen on routine brain MRI studies in controls older than 30 months of age and absent in all JS patients, confirming the proposed lack of commissural fibers in JS, and accounting in part for the molar tooth sign. PMID- 18412278 TI - Low-level mosaicism of trisomy 14: phenotypic and molecular characterization. AB - Trisomy 14 mosaicism is a rare cytogenetic abnormality with a defined and recognizable clinical phenotype. We present a detailed clinical history and physical findings of five patients with low-level mosaicism of trisomy 14 detected by array-based comparative genomic hybridization (array-CGH) analysis or by routine chromosome analysis. These patients exhibited growth and developmental delays with variable severity, congenital anomalies, pigmentary skin lesions, and dysmorphic features. The phenotype of our patients was compared with previously described cases. This report suggests that trisomy 14 mosaicism may be more common than has been previously appreciated and also illustrates the important application of array-CGH to detect low-level mosaic chromosome abnormalities. We predict that a wider application of the array-CGH technology will significantly increase the detection rate of low-level mosaicism and will subsequently improve our ability to provide a diagnosis for patients with dysmorphic features, congenital anomalies, and developmental delay. PMID- 18412279 TI - A novel mutation in NFKBIA/IKBA results in a degradation-resistant N-truncated protein and is associated with ectodermal dysplasia with immunodeficiency. AB - Alterations in nuclear factor kappa B (NF-kappaB) essential modulator (NEMO; HUGO approved symbol IKBKG) underlie most cases of ectodermal dysplasia with immune deficiency (EDI), a human disorder characterized by anhidrosis with diminished immunity. EDI has also been associated with a single heterozygous mutation at position Ser32 of the NF-kappaB inhibitor IkappaBalpha, one of two phosphorylation sites that are essential for targeting IkappaBalpha for proteasomal degradation and hence for activation of NF-kappaB. We report a novel heterozygous nonsense mutation in the IKBA (HUGO-approved symbol, NFKBIA) gene of a 1-year-old male child with EDI that introduces a premature termination codon at position Glu14. An in-frame methionine downstream of the nonsense mutation allows for reinitiation of translation. The resulting N-terminally truncated protein lacks both serine phosphorylation sites and inhibits NF-kappaB signaling by functioning as a dominant negative on NF-kappaB activity in lymphocytes and monocytes. These findings support the scanning model for translation initiation in eukaryotes and confirm the critical role of the NF-kappaB in the human immune response. PMID- 18412280 TI - Cerebellar ataxia, myoclonus, cervical lipomas, and MERRF syndrome. Case report. PMID- 18412281 TI - Microsubthalamotomy: an immediate predictor of long-term subthalamic stimulation efficacy in Parkinson disease. AB - A microsubthalamotomy (mSTN) effect is commonly described after implantation that improves Parkinson's motor disability and is considered to be an obvious sign of good placement of the definitive electrode within the subthalamic nucleus (STN). There has been no formal study, however, demonstrating whether this mSTN effect can predict the long-term efficacy of STN stimulation in Parkinson Disease. The mSTN effect was defined by the percentage improvement of unified parkinson's disease rating scale (UPDRS III) baseline score assessed the third day morning following STN implantation, after at least a 12 hour withdrawal of dopaminergic treatment and before the programmable pulse generator was switched on. It was assessed in 30 consecutive patients with PD submitted for STN stimulation. Multiple stepwise regression analysis showed that mSTN effect (P = 0.005) and global mean intensity of stimulation (P = 0.004) were accurate independent predictors of the 6-month postoperative UPDRS III motor score improvement in the off-drug/on-stimulation condition. PMID- 18412282 TI - rCBF changes associated with PPN stimulation in a patient with Parkinson's disease: a PET study. AB - Gait disturbances and akinesia are disabling symptoms in advanced Parkinson's disease (PD). The pedunculopontine nucleus (PPN) is involved in locomotion, control of posture, and behavioral states [i.e. wakefulness, rapid eye movement (REM) sleep]. Some reports have suggested that modulation of the activity of the PPN with deep brain stimulation (DBS) may be beneficial in the treatment of gait dysfunction and akinesia. To gain some insights on effects of PPN-DBS in the human brain, we used [(15)O] H(2)O positron emission tomography (PET) to measure changes in regional cerebral blood flow (rCBF) at rest during Off and On stimulation in an advanced PD patient with unilateral PPN-DBS. PPN-DBS increased rCBF in different subcortical areas most notably the thalamus, bilaterally. Double-blinded clinical evaluation revealed an improvement in motor function by approximately 20%. The PET changes provide for the first time evidence in the human brain that PPN-DBS may be able to influence and modify rCBF of closely connected subcortical structures. Given the importance of the PPN in locomotion, control of posture, and behavioral states, DBS may have significant implication for more complicated forms of movement disorders where deterioration of gait, postural instability, and REM sleep behavior disorders are very disabling. PMID- 18412283 TI - PET study of brain acetylcholinesterase in cerebellar degenerative disorders. AB - To elucidate characteristic changes of brain acetylcholinesterase (AChE) in cerebellar degenerative disorders. Eight patients with the cerebellar variant of multiple system atrophy (MSA-C), 7 patients with spinocerebellar ataxia type-3 (SCA-3), 3 patients with SCA-6, and 13 healthy age-matched volunteers participated in this study. Brain AChE activity was measured by [(11)C] N methylpiperidin-4-yl propionate PET in all subjects. Brain AChE activities were significantly decreased in the thalamus (-27%) and the posterior lobe of cerebellar cortex (-36%) in patients with MSA-C and in the thalamus (-23%) in patients with SCA-3 compared with healthy controls (P < 0.01). Thalamic AChE activities of SCA-3 patients were negatively correlated with the unified Parkinson's disease rating scale motor subscore (P < 0.001). AChE activities were not significantly altered in the cerebral cortex in any disease group. Reduction of AChE activities in the thalamus and cerebellum in MSA and in the thalamus in SCA-3 suggest that cholinergic modulating drugs may have a role in the treatment of ataxia and other symptoms in these disorders. PMID- 18412284 TI - Transcriptional expression of cis-acting and trans-acting splicing mutations cause autosomal dominant retinitis pigmentosa. AB - Two types of mutations may lead to deficient pre-mRNA splicing: cis-acting mutations that inactivate a constitutive or alternative splice site within the pre-mRNA, and trans-acting mutations that affect the function of a basal factor of the splicing machinery. Autosomal dominant retinitis pigmentosa (adRP) is caused by mutations in at least 12 genes, with mutations in rhodopsin being the most prevalent. Two cis-acting mutations, g.3811A>G and g.5167G>T at the splice site in the rhodopsin gene (RHO; GenBank U49742.1) are linked to adRP in a Spanish population; while a cis-acting mutation, g.4335G>T, has been linked to recessive RP (arRP). Transcriptional expression analysis showed that the cis acting splicing mutations linked to adRP promoted alternative splice sites, while the arRP linked mutation results in exclusion of exon 4. Trans-acting splicing mutations associated with adRP have also been found, and mutations in the pre mRNA splicing factors PRPF3, PRPF8, PRPF31, and RP9 are associated with adRP in several populations. This report describes a new mutation in PRPF3 in a Spanish adRP family. We also investigated the transcriptional patterns in Epstein-Barr virus (EBV)-transformed lymphoblastoid cells from patients carrying a mutation in PRPF8. Despite the role of PRPF8 in the minor U12 splicing processes, microarray analysis revealed that mutations in PRPF8 not only did not result in significant differences in splicing efficiency of rhodopsin, but no apparent changes in expression of U12-type intron genes and splicing processes was observed. Microarray analysis revealed a panel of differentially expressed genes mapped to the RP loci, and future work will determine their role in RP. PMID- 18412285 TI - Psychogenic propriospinal myoclonus. AB - We report a case of probable psychogenic propriospinal myoclonus (PSM) in a patient who developed a sudden onset of disabling axial flexor myoclonus following a cosmetic surgical procedure. The electrophysiological findings were consistent with previous reports of PSM. Spontaneous remissions and disappearance of the jerks, sustained for 2 years, following removal of superficial surgical screws support the diagnosis of a psychogenic movement disorder. PMID- 18412286 TI - RPS19 mutations in patients with Diamond-Blackfan anemia. AB - Diamond-Blackfan anemia (DBA) is an inherited disease characterized by pure erythroid aplasia. Thirty percent (30%) of patients display malformations, especially of the hands, face, heart, and urogenital tract. DBA has an autosomal dominant pattern of inheritance. De novo mutations are common and familial cases display wide clinical heterogeneity. Twenty-five percent (25%) of patients carry a mutation in the ribosomal protein (RP) S19 gene, whereas mutations in RPS24, RPS17, RPL35A, RPL11, and RPL5 are rare. These genes encode for structural proteins of the ribosome. A link between ribosomal functions and erythroid aplasia is apparent in DBA, but its etiology is not clear. Most authors agree that a defect in protein synthesis in a rapidly proliferating tissue, such as the erythroid bone marrow, may explain the defective erythropoiesis. A total of 77 RPS19 mutations have been described. Most are whole gene deletions, translocations, or truncating mutations (nonsense or frameshift), suggesting that haploinsufficiency is the basis of DBA pathology. A total of 22 missense mutations have also been described and several works have provided in vitro functional data for the mutant proteins. This review looks at the data on all these mutations, proposes a functional classification, and describes six new mutations. It is shown that patients with RPS19 mutations display a poorer response to steroids and a worse long-term prognosis compared to other DBA patients. PMID- 18412287 TI - Poly(ether ester amide) microspheres for protein delivery: influence of copolymer composition on technological and biological properties. AB - The production of PEEA microspheres with potential as carriers for protein oral delivery is described. PEEAs with different hydrophilicity were synthesized and characterized. Experiments showed that an increase in copolymer hydrophilicity gave particles less prone to cell interaction. BSA release profiles from PEEA microspheres demonstrated that an increase in polymer hydrophilicity was useful in limiting protein burst and modulating drug delivery rate by increasing PEEA degradability. These results show that fine-tuning of the hydrophilic/hydrophobic properties of PCL is essential for the formulation protein-loaded microspheres with specific properties. PMID- 18412288 TI - Novel types of carborane-carrier hyaluronan derivatives via "click chemistry". AB - Two new HA derivatives bearing carborane rings were synthesized by click chemistry. The optimal conditions were assessed for the preparation of biocompatible boron carriers, potentially suitable for application in BNCT and capable of targeting the CD44 antigen. The new polymeric samples were characterized by means of NMR-spectroscopy techniques that gave degrees of 17 and 8% for HAAACB and HapACB, respectively. Both HAAACB and HApACB turned out to be nontoxic for colorectal, ovarian and bladder tumor cell lines, to disclose a specific interaction with the CD44 antigen as the native hyaluronan moiety, and to deliver boron-atom concentrations largely sufficient for BNCT therapy when accumulated in cancer cells. PMID- 18412289 TI - Diagnostic flow cytometry for low-grade myelodysplastic syndromes. AB - It has long been considered that flow cytometry (FCM) has little role in clinical practice in the diagnosis of myelodysplastic syndromes (MDS). However, recent advances in the analytical method and knowledge of MDS FCM are changing this stereotype. This paper reviews the concept and current status of FCM in the diagnosis of low-grade MDS. The diagnosis of low-grade MDS in the absence of ringed sideroblasts and chromosomal aberration is not always straightforward, and a report from a recent international working conference has proposed FCM as an adjunctive diagnostic test for such cases. Currently, only a limited number of laboratories are applying FCM to the diagnosis of MDS. Furthermore, standard analytical methods in FCM for MDS have not been established, and no single FCM parameter is sufficiently sensitive and specific to make the diagnosis of MDS. To establish MDS FCM as a widely accepted, dependable diagnostic tool, prospective studies should increase flow parameters that can be analysed reproducibly and determine their sensitivity and specificity, either alone or in combination. CD34+ cell-related parameters that are applicable for diagnosing low-grade MDS in many laboratories are introduced here. PMID- 18412290 TI - Chronic statin therapy and the risk of colorectal cancer. AB - BACKGROUND AND AIMS: Epidemiologic studies on a potential chemopreventive effect of statin therapy have yielded conflicting results. We sought to clarify whether long-term statin therapy has a chemopreventive effect on the risk of colorectal cancer (CRC) in a large, population-representative cohort. METHODS: A nested case control study was conducted among patients > or =50 years of age and with > or =5 years of CRC-free initial follow-up in the General Practice Research Database (GPRD; 1987-2002). Cases consisted of all patients with incident CRC. Up to 10 controls were matched with each case on practice site and both duration and calendar time of follow-up prior to the index date. The primary exposure of interest was > or =5 years of cumulative statin use. RESULTS: We identified 4432 incident CRC cases and 44 292 controls. The adjusted odds ratio (OR) for > or =5 years of statin exposure was 1.1 (95% confidence interval (CI): 0.5-2.2). Chronic NSAID/aspirin use did not modify this primary association (test for interaction, p = 0.5). Compared to statin non-users, the adjusted OR for 10 years of statin exposure was 1.3 (95% CI: 0.6-2.7), and the adjusted OR associated with the highest quartile of cumulative statin dose was 1.2 (95% CI: 0.9-1.7). There was a non-statistically significant trend towards a possible reduction in CRC risk among users of high daily statin dose. CONCLUSION: Long-term statin therapy at usual doses was not associated with a significantly reduced risk of CRC. A chemopreventive effect at high daily doses cannot be excluded. PMID- 18412292 TI - Peritoneal lavage cytology in patients with oesophagogastric adenocarcinoma. AB - BACKGROUND: The aim of the study was to determine the value of performing peritoneal lavage cytology during laparoscopy in the management of oesophagogastric adenocarcinoma. METHODS: Laparoscopy combined with peritoneal cytology was performed in patients with potentially resectable oesophagogastric adenocarcinoma. Macroscopic peritoneal findings at laparoscopy and the presence of free peritoneal tumour cells were recorded. All patients were followed to death or the census point. Patients with overt peritoneal disease or positive cytology were offered palliative chemotherapy, subject to performance status. RESULTS: Forty-eight (18.8 per cent) of 255 patients had overt peritoneal metastases at staging laparoscopy. Fifteen (7.2 per cent) of the remaining 207 patients had positive cytology; these patients had a median (95 per cent confidence interval) survival of 13 (3.1 to 22.9) months, versus 9 (7.4 to 10.6) months for those with overt peritoneal metastases (P = 0.517). Of patients receiving chemotherapy, those without overt metastases had a slight survival advantage over patients with metastases (median 15 (10.8 to 19.2) versus 9 (7.4 to 10.7) months; P = 0.045). CONCLUSION: Positive peritoneal cytology in the absence of overt peritoneal metastases is not uncommon in oesophagogastric adenocarcinoma. It is a marker of poor prognosis even in the absence of overt peritoneal metastases. PMID- 18412291 TI - The MRI brain correlates of depressed mood, anhedonia, apathy, and anergia in older adults with and without cognitive impairment or dementia. AB - OBJECTIVES: We examined the magnetic resonance imaging (MRI) correlates of depressed mood, apathy, anhedonia, and anergia in older adults with and without cognitive impairment or dementia. METHODS: This analysis included 270 community dwelling older adults (59% male; 79% Caucasian; mean age 74.4 years) who were recruited into a multi-center longitudinal observational study of subcortical ischemic vascular disease (SIVD).The distribution of cognitive status included: cognitively intact (38%), cognitively impaired (27%), or demented (35%). All subjects underwent MRI and 41% were classified as having subcortical lacunes. MRI measures included cortical gray and white matter volumes, lacunar volumes in subcortical white and gray matter structures, volume of white matter hyperintensities, and total hippocampal volume. Depressed mood, anhedonia, anergia, and apathy apparent at the time of assessment were assessed using a behavioral assessment Associations between neuropsychiatric symptoms and MRI variables were evaluated using logistic regression. RESULTS: Subjects with neuropsychiatric symptoms were more likely to be cognitively impaired or demented than those without neuropsychiatric symptoms. In multivariate models controlling for cognitive status, age, gender, and education, higher lacunar volume in white matter was independently associated with the presence of all four neuropsychiatric symptoms. CONCLUSIONS: We report an association between the lacunar volumes in the white matter and depressed mood, anhedonia, apathy, and anergia, thus supporting the role of subcortical ischemic vascular disease in the pathogenesis of late-life neuropsychiatric disorders. PMID- 18412293 TI - Sacral nerve stimulation for faecal incontinence alters colorectal transport. AB - BACKGROUND: Sacral nerve stimulation reduces the frequency of defaecation in patients with faecal incontinence. The aim of this study was to examine the mechanism behind the reduced number of bowel movements in incontinent patients treated with sacral nerve stimulation. METHODS: The study included 20 patients with faecal incontinence and a positive percutaneous nerve evaluation test: 19 women and one man, with a median age of 63 (range 28-78) years. Colorectal scintigraphy was performed to assess colorectal emptying at defaecation before and after implantation. Segmental colorectal transit times were determined using radio-opaque markers. RESULTS: The median frequency of defaecation per 3 weeks decreased from 56 (range 19-136) to 26 (range 12-78) (P < 0.002). At defaecation, antegrade transport from the ascending colon decreased from a median score of 8 (range 0-23) to 0 (range 0-11) per cent (P = 0.001), while retrograde transport from the descending colon increased from a median score of 0 (range 0-14) to 2 (range 0-30) per cent (P = 0.039). The median defaecation score was unchanged. There was a non-significant increase in median total gastrointestinal transit time from 2.5 (range 0.9-6.2) to 3.3 (range 0.8-6.2) days (P = 0.079). CONCLUSION: Sacral nerve stimulation reduces antegrade transport from the ascending colon and increases retrograde transport from the descending colon at defaecation. This may prolong colonic transit time and increase the storage capacity of the colon. PMID- 18412294 TI - Risk factors for complications after partial splenic embolization for liver cirrhosis. AB - BACKGROUND: Partial splenic embolization (PSE) in patients with cirrhosis can achieve a prolonged increase in blood cell count. However, there is little information on the risk factors for complications after PSE for liver cirrhosis. METHODS: Seventy patients (41 men and 29 women) with cirrhosis-induced hypersplenism underwent PSE. Based on the Child-Pugh classification, 30, 35 and five patients had class A, B and C disease respectively. Risk factors for complications after PSE were examined retrospectively by logistic regression analysis. RESULTS: The mean splenic infarction rate was 77.1 per cent. Twelve patients (17 per cent) had complications following PSE. In univariable analysis, Child-Pugh class C, total bilirubin level of 1.4 mg/dl or more, serum albumin level of 2.8 g/dl or less and infarcted splenic volume of 540 ml or more were significantly associated with complications after PSE. In multivariable analysis, Child-Pugh class C (P = 0.012) and infarcted splenic volume of 540 ml or more (P = 0.031) were identified as risk factors, with an odds ratio of 22.92 and 5.01 respectively. CONCLUSION: Child-Pugh class C and a large splenic infarction volume are risk factors for complications after PSE for liver cirrhosis. PMID- 18412295 TI - Cost-effectiveness of endarterectomy in patients with asymptomatic carotid artery stenosis. AB - BACKGROUND: Long-term health outcomes and costs are important when deciding whether a strategy of carotid endarterectomy in addition to best medical management should be recommended for patients with asymptomatic carotid artery stenosis. This study investigated the cost-effectiveness of such a strategy compared with a strategy of best medical management alone. METHODS: Based on data from the randomized Asymptomatic Carotid Surgery Trial (ACST), a national vascular database and other published sources, expected costs and health outcomes in terms of quality-adjusted life years (QALYs) of both treatment strategies were estimated using decision-analytical modelling. Cost-effectiveness was established for a Swedish setting from a societal perspective. RESULTS: Base-case analysis showed that the incremental cost per QALY of a strategy with carotid endarterectomy for 65- and 75-year-old men (women) was 34,557 euros (311,133 euros) and 58,930 euros (779,776 euros) respectively. Sensitivity analyses indicated that the duration of the treatment effect after 5 years of follow-up in the ACST was important for the cost-effectiveness results. CONCLUSION: Carotid endarterectomy in addition to best medical management can be considered cost effective in men aged 73 years or less but is less likely to be cost-effective in older men or in women. PMID- 18412296 TI - Nitric oxide reduces organ injury and enhances regeneration of reduced-size livers by increasing hepatic arterial flow. AB - BACKGROUND: Reduced-size livers suffer from portal hyperperfusion, diminished arterial blood flow and the risk of postoperative liver injury. The aim of this experimental study was to unravel the role of nitric oxide in this setting. METHODS: Rats underwent 85 per cent partial hepatectomy and either substitution of nitric oxide with molsidomine or inhibition of nitric oxide synthase (NOS) with N(G)-nitro-L-arginine methyl ester. Untreated hepatectomized animals served as controls and unresected animals as the sham group. RESULTS: Ultrasonic flowmetry following partial hepatectomy revealed a marked increase in portal venous inflow with a concomitant decrease in hepatic arterial inflow. Nitric oxide substitution counteracted the decline in hepatic arterial inflow and caused a significantly greater increase in cell proliferation after partial hepatectomy compared with control or NOS-inhibited animals. Hepatectomized animals further profited from nitric oxide substitution, as indicated by reduced aminotransferase release and improved liver function. CONCLUSION: Nitric oxide improves the postoperative course of rats with reduced-size livers by modulating hepatic macrohaemodynamics and mediating regeneration and cytoprotection, but not by reducing hepatic hyperperfusion and the accompanying sinusoidal shear stress. PMID- 18412297 TI - Outcome of younger patients with acute diverticulitis. AB - BACKGROUND: There is controversy over whether patients presenting with a primary attack of acute diverticulitis at a younger age are more prone to complications and recurrence than older patients. METHODS: A review, including postal questionnaires, was undertaken of 234 patients who had a primary episode of acute diverticulitis. The diagnosis was confirmed by computed tomography (CT) and/or pathology report. The mean length of follow-up was 30 (range 16-45) months. RESULTS: In 58 patients aged 50 years or less no differences in fever or white blood cell count were found in comparison with findings in 176 patients aged above 50 years. The rate of severe diverticulitis observed with CT was lower in the younger patients (2 versus 11.9 per cent; P = 0.025). Surgical management during the first admission was undertaken less commonly in younger patients (2 versus 6.8 per cent; P = 0.271). Rates of subsequent events (recurrent diverticulitis and/or further surgery) during follow-up were higher in younger patients (25 versus 19.5 per cent), but this was not significant (P = 0.423). A type II error cannot be excluded. CONCLUSION: First episodes of acute diverticulitis were not more aggressive in patients aged 50 years or less. Recurrence rates were slightly higher than in older patients. PMID- 18412298 TI - Cutaneous granulomas during infliximab therapy for spondyloarthropathy. PMID- 18412299 TI - Chikungunya viral polyarthritis. PMID- 18412300 TI - Arthritis in Aboriginal Manitobans: evidence for a high burden of disease. AB - OBJECTIVE: To evaluate the relative burden of arthritis and patterns of care in Aboriginal Manitobans, using multiple data sets to ensure a representative picture. METHODS: Arthritis burden and healthcare utilization was ascertained using 3 separate data sources. Physician claims for 3 common ICD-9 musculoskeletal diagnoses were abstracted from the Population Health Research Data Repository for First Nations (FN) Manitobans and compared to all other Manitobans. Self-reported arthritis rates were obtained from the Manitoba First Nations Regional Longitudinal Health Survey (MFN Survey), which surveyed FN persons living on-reserve. Data on ethnicity and diagnoses were abstracted from the Arthritis Centre research database, which contains records of all patients seen at the Arthritis Centre. RESULTS: Twice as many FN Manitobans had physician claims for rheumatoid arthritis, degenerative arthritis, and unspecified arthropathy compared to all other Manitobans. MFN Survey data identified a self reported arthritis rate of 21.0% and a rheumatoid arthritis (RA) rate of 3.0%. Data for 687 Aboriginal patients and 4135 Caucasian patients were abstracted from the Arthritis Centre database. Aboriginal patients seen in the Arthritis Centre were 2 to 4 times more likely to have a diagnosis of inflammatory disease, and less than half as likely to have noninflammatory disease. CONCLUSION: The data highlight the increased burden of arthritis in Aboriginal Manitobans, and draw attention to large gaps in our knowledge of how, why, and when Aboriginals access medical care. PMID- 18412301 TI - Patient pain and tissue trauma during syringe procedures: a randomized controlled trial. AB - OBJECTIVE: To investigate the relationship of needle control to tissue trauma and hemorrhage during syringe procedures. METHODS: Forty-seven subjects with a palpable knee effusion underwent needle and syringe aspiration. Subjects were randomized to the conventional syringe or a safety technology, the reciprocating procedure device (RPD). This trial was registered at clinicaltrials.gov. Pain was measured with the Visual Analog Pain Scale (VAPS). Cell count, crystal examination, culture, and aspirated fluid volume were determined. Red blood cell (RBC) counts were used to measure blood in aspirated fluid. RESULTS: Patient pain during the syringe procedure significantly predicted blood (RBC) in aspirated fluid (r = 0. 53, p = 0.001). When compared to the conventional syringe, the RPD safety device reduced blood in aspirated fluid by 66.7% (RBC, 10(3)/ml: RPD 8.9 +/- 11.4; syringe 26.7 +/- 90.2; p 0.01), reduced patient pain by 73.9% (VAPS: RPD 1.68 +/- 2.34; syringe 6.44 +/- 2.86; p < 0.01), and improved fluid aspirate yield by 132% (aspirate volume: RPD 20.9 +/- 19.7 ml; syringe 9.00 +/- 6.58 ml; p < 0.01). CONCLUSION: Inadequate control of needle and syringe during physician performed syringe procedures is an important cause of trauma to patient tissues resulting in hemorrhage, increased patient pain, and decreased aspirate yield. The RPD -- a safety device that improves needle control and decreases needle trauma to tissues -- reduces hemorrhage and improves the safety, outcome, and aspirate yield of physician-performed syringe procedures. PMID- 18412302 TI - Association between crystals and cartilage degeneration in the ankle. AB - OBJECTIVE: Monosodium urate (MSU) and calcium pyrophosphate dihydrate (CPPD) crystals have been observed in synovial joints both before and after the onset of osteoarthritis (OA). The relationship between crystals and OA, however, remains controversial. We compared histologic and immunohistochemical patterns in articular cartilage of ankle joints with and without crystals. METHODS: A sample of 7,855 human cadaveric tali was examined for the presence of surface and beneath-the-surface crystals. A random subsample of tali with and without crystals underwent crystal analysis by Fourier transform infrared spectroscopy (FTIR), histologic examination, and immunohistochemistry for S100 protein, superficial zone protein, collagen X, cSRC. RESULTS: The prevalence of grossly visible crystals in the pool of donors over 18 years of age was 4.7% and correlated with advanced age, male sex, and obesity. Crystals were strongly associated with cartilage lesions and these lesions appeared to be biomechanically induced, being located where opposing articular surfaces might not be in congruence with each other. Thirty-four percent of the random subsamples of crystals upon which FTIR was performed contained CPPD, and the remainder were MSU crystals. Both crystal types were associated with higher levels of superficial zone protein and collagen X. CONCLUSION: We show that the presence of surface crystals of either MSU or CPPD is strongly correlated with cartilage lesions in the talus. The histologic similarities in cartilage from joints with CPPD crystals compared to those with MSU crystals, together with what is known about the dramatically different etiologic factors producing these crystals, strongly suggest that these lesions are biomechanically induced. PMID- 18412303 TI - Evidence of silicon dioxide crystals in synovial fluid of patients with osteoarthritis. AB - OBJECTIVE: Synovial fluid (SF) may contain a number of crystals that optical microscopy is unable to identify with certainty. Scanning electron microscopy (SEM) was utilized in this study to characterize SF crystals in the context of knee osteoarthritis (OA). METHODS: SF was collected from the knees of 25 patients with OA and examined under optical light microscopy. Calcium pyrophosphate dihydrate (CPPD) crystals were assessed by means of compensated polarized light microscopy, while alizarin red S staining was performed to identify apatite (BCP) crystals. All the specimens were also analyzed by SEM and x-ray diffractometry, as gold standards. RESULTS: CPPD crystals were found in 32% and BCP in 24% of the SF examined by SEM. The degree of concordance between polarized light microscopy and SEM was 0.83 for CPPD and 0.46 for BCP (kappa statistic). The secondary and backscatter electron SEM observations allowed identification of silicon dioxide (SiO2) crystals in 8 out of 10 patients in whom polarized light microscopy revealed irregular and polymorph crystals. CONCLUSION: SiO2 crystals cannot be readily identified by their morphology or polarization properties under optical microscopy. Their presence, nevertheless, did not lead to misclassification. PMID- 18412304 TI - Correlation between inflammatory cells and sulfated glycosaminoglycan concentration in synovial fluid of subjects with secondary knee osteoarthritis. AB - OBJECTIVE: To analyze the presence of inflammatory cells in the synovial fluid (SF) of subjects with osteoarthritis (OA) and correlate them with the concentration of sulfated glycosaminoglycan (GAG). METHODS: The degree of chondral injury from 20 subjects (17 men, 3 women, mean age 48.06 +/- 14.35 yrs) was evaluated macroscopically by means of arthroscopy and graded according to the International Cartilage Repair Society classification. All subjects presented anterior cruciate ligament (ACL) injuries, associated or not to menisci injuries. SF was aspirated and centrifuged. Sulfated GAG concentration was quantified by dimethylene blue staining. Manual morphometry was used to determine the inflammatory cell count (mononuclear and polymorphonuclear) by optical microscopy, after staining with May-Grunwald-Giemsa. The time of injury and degree of chondral injury were considered, and correlations among the variables were obtained by Spearman nonparametric correlation test. RESULTS: There was no significant correlation between the amount of mononuclear cells and the GAG concentration. There was a significant positive correlation between the polymorphonuclear cells and the GAG concentration, and a significant negative correlation between the degree of chondral injury and the GAG concentration. CONCLUSION: Polymorphonuclear cells may be either a cause or an effect in the metabolic and biochemical processes occurring in chondral injuries. The correlations support the hypothesis that inflammatory cells have a significant role in the progression and chronicity of chondral injury in secondary OA. PMID- 18412305 TI - Patients with scleroderma may have increased risk of osteoporosis. A comparison to rheumatoid arthritis and noninflammatory musculoskeletal conditions. AB - OBJECTIVE: To investigate if subjects with scleroderma (systemic sclerosis, SSc) have increased risk for developing osteoporosis (OP). METHODS: A survey assessing demographics, diagnosis/investigations for OP, and risk factors for OP was mailed to 129 patients with SSc, 158 controls with noninflammatory musculoskeletal (MSK) disease, and 230 positive controls with rheumatoid arthritis (RA). All available charts were reviewed and results were included in analyses of demographics, OP status, past bone mineral density (BMD), and past steroid use. In addition, we recorded BMD results (T score) of SSc patients with their matched RA controls. Analyses adjusted for age were done for SSc versus MSK and SSc versus RA. RESULTS: The response rate was 61% for patients with SSc (n = 28 diffuse, 51 limited disease), RA 67%, and MSK 59%; however, through chart review, 159 SSc, 140 MSK, and 235 RA patients were included in the analyses. Mean age and proportion of women did not differ between groups. Disease duration was longer in RA versus SSc group (16.5 vs 11.5 yrs; p < 0.0001). The prevalence of OP in SSc was similar to RA controls (19.4% vs 16.7%; p = 0.38) but likely higher than MSK controls (12.2%; p = 0.054). Subjects with SSc reported a higher rate of disability (41.0% vs 15.6%; p = 0.0001) and less family history of OP (22.8% vs 46.7%; p = 0.0006) compared with the MSK control group. There were no differences between groups in reports of fracture (35% SSc, 43% MSK, 37% RA; p = 0.5) or OP related fractures (4% SSc, 11% MSK, 11% RA; p = 0.5). Subjects with SSc were less likely to have had a BMD done in the past compared to RA (40.9% vs 62.6%; p = 0.0001). Subjects with RA who reported OP had longer disease duration than RA without OP (18 +/- 1.7 yrs vs 12 +/- 0.8; p = 0.0009). Results from the chart review showed that the T scores of SSc (n = 56, mean age 62.9 +/- SD 10.1 yrs) at lumbar spine (SSc -1.01 vs RA -0.97), femoral neck (SSc -2.07 vs RA -1.46; p = 0.01, adjusting for age p = 0.26), and total hip region (SSc -1.52 vs RA -1.25) were comparable to or even lower than the RA group (n = 56, mean age 62.2 +/- SD 10.7 yrs). CONCLUSION: The prevalence of OP in patients with SSc was comparable to those with RA, but higher than in the MSK group. Age was found to be an important factor, as expected. Also, our results indicated that BMD (T score) in SSc was similar to or even lower than in patients with RA. Increasing the awareness to order BMD measurements in patients with SSc may be warranted based on our results, especially for older patients. PMID- 18412306 TI - Longterm visual prognosis of patients with ocular Adamantiades-Behcet's disease treated with interferon-alpha-2a. AB - OBJECTIVE: Relapsing ocular involvement is one of the major manifestations in Adamantiades-Behcet's disease (ABD). Combining systemic corticosteroids with cyclosporin A is currently the treatment of choice. Interferon-alpha (IFN-alpha) has proven to be effective in mucocutaneous ABD and has been reported to improve ocular lesions. We examined the longterm effects of IFN-alpha-2a in a case series of 45 patients with ocular involvement. METHODS: Since 1988, 45 patients (79 eyes of 90 eyes) with ocular involvement in ABD have been treated with IFN-alpha (3 x 6-9 Mio IU per wk). In the initial acute phase of the disease, patients additionally received short-term corticosteroids (oral prednisolone 100 mg/day), tapered to a maintenance dose of 10 mg/day within 2 weeks. IFN-alpha-2a was administered as longterm therapy with a mean duration of 30 months (range 1.1-101 mo). RESULTS: IFN-alpha-2a/prednisolone treatment was effective against vasculitis, optic nerve neuropathy, and iritis. Sixty-four eyes had no recurrence under therapy. To date, recurrences have been seen in 26 eyes under IFN-alpha treatment. Flu-like symptoms were recorded in nearly all patients (n = 43). Further side effects were dose-dependent reversible thrombocytopenia (n = 1), psychosis (n = 3), depression (n = 13), thyroiditis (n = 1), and reversible diffuse alopecia (n = 7). In our series, 92% of all eyes showed stable or improved visual acuity in longterm followup. CONCLUSION: Longterm remission of ocular inflammation can be achieved with the combination of IFN-alpha and low dose corticosteroids. PMID- 18412308 TI - HLA-DRB1 alleles and Henoch-Schonlein purpura: susceptibility and severity of disease. AB - OBJECTIVE: The genetic basis of susceptibility to Henoch-Schonlein purpura (HSP) may be conferred by a number of gene loci, including the MHC. Associations between human leukocyte antigen (HLA) and disease can help to establish a basis for susceptibility and assist in the prediction of the outcome and clinical heterogeneity. We aimed to investigate the implications of the HLA-DRB1 locus and the susceptibility to HSP, and to determine if there are associations with joint, gastrointestinal, and renal manifestations of the disease. METHODS: We studied 110 Turkish patients (men/women: 66/44) with HSP. Patients and ethnically matched controls with respect to age and sex (n = 250) were HLA-DRB1 genotyped from DNA determined using molecular based methods. RESULTS: HLA-DRB1 genotype differences between patients with HSP and controls were observed. The frequency of HLA-DRB1 11/14 was higher [odds ratio (OR) 1.97, 95% confidence interval (95% CI) 1.25 3.12, p = 0.003; OR = 1.83, 95% CI = 1.02-3.28, p = 0.035, respectively] and the frequency of HLA-DRB1 10/17 was lower (OR = 1.04, 95% CI = 1.01-1.86, p = 0.035; OR = 3.96, 95% CI = 1.17-13.33, p = 0018, respectively) in patients with HSP compared to controls. No HLA-DRB1 associations with gastrointestinal and renal manifestations were found (p > 0.05). In contrast, HLA-DRB1*11 positivity was increased and HLA-DRB1 14 positivity reduced in HSP patients with joint manifestations (OR = 2.68, 95% CI = 1.09-6.66, p = 0.029; OR = 9.34, 95% CI = 3.38-25.64, p = 0.000, respectively). Also, HLA-DRB1 13 positivity was found to be increased in patients with nephrotic proteinuria (OR = 3.76, 95% CI = 1.25 11.23, p = 0.025). CONCLUSION: These results suggest that genetic factors from HLA-DRB1 genotypes might be related to the susceptibility to HSP for Turkish children but not to the severity of this disease. Additional studies are required to confirm the association of alleles encoded in the HLA region with the disease progression and severity. PMID- 18412309 TI - TCRBV20S1 and TCRBV3S1 gene segment polymorphisms in systemic sclerosis. AB - OBJECTIVE: To compare the frequencies of variants of TCRBV20S1 and TCRBV3S1 gene segments in patients with systemic sclerosis (SSc) and in controls. The null allele (allele 2) of TCRBV20S1 is associated with reduced levels of Vbeta20+ T cells in the peripheral blood, while allele 1 of TCRBV3S1 is related to a low frequency of Vbeta3.1+ T-cells. METHODS: One hundred thirty patients with SSc and 118 healthy volunteer controls were genotyped for TCRBV20S1, and 117 patients and 85 controls were genotyped for TCRBV3S1 variants by PCR-RFLP. Patients underwent clinical evaluation, serology, pulmonary function tests, high resolution computed tomography, and Doppler echocardiography. RESULTS: The genotypic frequencies of TCRBV20S1 were 0.46 (allele 1/allele 1), 0.43 (allele 1/allele 2), and 0.11 (allele 2/allele 2) in SSc patients; in controls the frequencies were 0.70, 0.26, and 0.04, respectively (p < 0.001). The Mantel-Haenszel odds ratio (stratified by race and sex) of the allele 2 carrier state was 3.88 (95% CI 1.94 to 7.75). The allelic and genotypic frequencies of the TCRBV3S1 gene segment did not differ significantly in patients and controls. However, among patients, allele 1 (TCRBV3S1) carriers had a higher prevalence of interstitial lung disease (adjusted p = 0.032). CONCLUSION: The null allele of the TCRBV20S1 and the allele 1 of TCRBV3S1 gene segments may be considered risk factors for the development of SSc and interstitial lung disease, respectively, suggesting a protective role of Vbeta20+ and Vbeta3.1+ cells in the pathogenic immune responses in SSc. PMID- 18412310 TI - Palindromic rheumatism is a common disease: comparison of new-onset palindromic rheumatism compared to new-onset rheumatoid arthritis in a 2-year cohort of patients. AB - OBJECTIVE: To determine the prevalence of palindromic rheumatism (PR) compared to new-onset rheumatoid arthritis (RA). METHODS: We reviewed 145 patients that had been newly diagnosed by a rheumatologist with either RA or PR between May 2004 and May 2006. RESULTS: Of these 145 patients, 51 were diagnosed with PR and 94 with RA. There was a similar female predominance with both conditions. The average age at diagnosis of PR was 49 years as compared to 56 years for RA. CONCLUSION: Palindromic rheumatism occurs more frequently than previously recognized. PMID- 18412311 TI - The problem of rheumatoid arthritis disease activity and remission in clinical practice. AB - OBJECTIVE: To investigate the results and feasibility of available scales to measure minimal disease activity (MDA) and remission in rheumatoid arthritis (RA) in the clinic. METHODS: We studied 849 consecutive patients with RA seen in a community rheumatology practice for routine RA care by 4 rheumatologists, beginning in March 2007 and ending in August 2007. Patients and physicians completed a simple form at each visit. We calculated the Disease Activity Score 28 (DAS28), Clinical Disease Activity Index (CDAI), physician assessment of global activity, and the Patient Activity Scale (PAS-II). From these we calculated remission and MDA prevalence in this community practice. RESULTS: The DAS28 could not be determined in more than 50% of patients because of referring physician and insurance company restrictions. Remission prevalences differed by assessment method: DAS28 28.5%, CDAI 6.5%-8.1%, physician global 12.5%, PAS 13.7%. MDA was 26.9% using the American College of Rheumatology core set variables, 34.7% using the DAS28, and 26.8% using the PAS-II. The kappa statistic was only fair (0.2 to 0.4) for most comparisons between assessment methods. No significant differences were noted for remission and MDA according to biologic therapy. CONCLUSION: The CDAI and/or physician global and PAS-II are simple acceptable ways to measure RA activity in the clinic, but results differ strikingly according to method. Further standardization appears to be required for full implementation of the CDAI. Caution is urged before using these methods for regulatory purposes. PMID- 18412312 TI - Mortality trends in rheumatoid arthritis: the role of rheumatoid factor. AB - OBJECTIVE: We previously demonstrated a widening in the mortality gap between subjects with rheumatoid arthritis (RA) and the general population. We examined the contribution of rheumatoid factor (RF) positivity on overall mortality trends and cause-specific mortality. METHODS: A population-based RA incidence cohort (1955-1995, and aged >or= 18 yrs) was followed longitudinally until death or January 1, 2006. The underlying cause of death as coded from national mortality statistics and grouped according to ICD-9/10 chapters was used to define cause specific mortality. Expected cause-specific mortality rates were estimated by applying the age-, sex-, and calendar-year-specific mortality rates from the general population to the RA cohort. Poisson regression was used to model the observed overall and cause-specific mortality rates according to RF status, accounting for age, sex, disease duration, and calendar year. RESULTS: A cohort of 603 subjects (73% female; mean age 58 yrs) with RA was followed for a mean of 16 years, during which 398 died. Estimated survival at 30 years after RA incidence was 26.0% in RF+ RA subjects compared to 36.0% expected (p < 0.001), while in RF- RA subjects, estimated survival was 29.1% compared to 28.3% expected (p = 0.9). The difference between the observed and the expected mortality in the RF+ RA subjects increased over time, resulting in a widening of the mortality gap, while among RF- RA subjects, observed mortality was very similar to the expected mortality over the entire time period. Among RF+ RA subjects, cause specific mortality was higher than expected for cardiovascular [relative risk (RR) 1.50; 95% confidence interval (CI) 1.22, 1.83] and respiratory diseases [RR 3.49; 95% CI 2.51, 4.72]. Among RF- RA subjects, no significant differences were found between observed and expected cause-specific mortality. CONCLUSION: The widening in the mortality gap between RA subjects and the general population is confined to RF+ RA subjects and largely driven by cardiovascular and respiratory deaths. PMID- 18412314 TI - Low-dose prednisolone in rheumatoid arthritis: adverse effects of various disease modifying antirheumatic drugs. AB - OBJECTIVE: To assess the incidence and severity of disease modifying antirheumatic drug (DMARD)-induced adverse effects (AE) in patients with rheumatoid arthritis (RA) taking/not taking glucocorticoids (GC). More specifically, we tested whether GC can prolong the survival time of DMARD in patients receiving combination therapy. METHODS: In a retrospective study of 154 patients with RA, data were examined for DMARD therapy and duration of low-dose GC ((3/4) 7.5 mg prednisone equivalent/day). Patients were followed for 2-62 months, and AE were graded following WHO criteria. RESULTS: GC therapy significantly increased the duration of therapy with sulfasalazine (SSZ) from 10.4 +/- 2.3 to 22.5 +/- 1.9 months and for methotrexate (MTX) from 21.8 +/- 2.9 to 43.3 +/- 2.7 months. Stratifying the withdrawal of DMARD for occurrence of AE and loss of efficacy revealed that GC comedication significantly increased the time until AE for users of MTX (3.0 +/- 0.6 vs 18.8 +/- 1.3 mo; p < 0.05), hydroxychloroquine (HCQ; 34.5 +/- 4.6 vs 54.4 +/- 5.1 mo; p < 0.05), and gold (6.6 +/- 0.9 vs 10.5 +/- 0.9 mo; p < 0.05). In patients taking SSZ the time until cessation due to loss of efficacy increased significantly under GC comedication (16.8 +/- 1.2 vs 31.3 +/- 2.9 mo; p < 0.05). However, in patients taking azathioprine (AZA) the duration of therapy decreased from 44.4 +/- 2.6 to 22.3 +/ 1.6 months under GC due to both time until AE and loss of efficacy. Patients under comedication of MTX + GC, HCQ + GC, and AZA + GC experienced significantly more AE compared to the respective DMARD monotherapy. A highly significant reduction was observed in the frequency of erosive RA in patients with GC comedication (n = 30; 49.1%) compared to patients without low-dose GC (n = 81, 80.4%; OR 4.05, 95% CI 1.91-8.66, p < 0.0001). CONCLUSION: Low-dose GC retard radiological progression of RA and exhibit a differential effect on survival of DMARD and degree of AE due to DMARD. Further studies are warranted to address safety and interactions of chronic low-dose GC in RA patients treated with DMARD. PMID- 18412313 TI - Relative responsiveness of physician/assessor-derived and patient-derived core set measures in rheumatoid arthritis trials. AB - OBJECTIVE: We assessed whether individual American College of Rheumatology core set measures (CSM), and the CSM grouped as composite patient-derived (CPD) or composite physician/assessor-derived (CMD), performed differently in rheumatoid arthritis (RA) clinical trials. METHODS: We used data from 9 RA trials [anti tumor necrosis factor-alpha (TNF-alpha) and disease modifying antirheumatic drug (DMARD)] in which CSM had been assessed, conducted from the early 1990s to present, with a total of 2969 patients. We grouped the CSM as CPD (pain, patient global assessment, function) and CMD [tender joint count (TJC), swollen joint count (SJC), physician global, inflammatory marker]. Using bootstrap simulation, we estimated the sample size that would be required to distinguish active treatment from placebo with the Wilcoxon rank-sum test in the clinical trials for the outcomes of percentage change of each individual CSM, of the Disease Activity Score (DAS), and average percentage change of the CMD or of the CPD. RESULTS: Comparing the performance of individual CSM relative to one another, the physician and patient global assessments and TJC would require the lowest sample sizes to distinguish active treatment from placebo, while use of the SJC, inflammatory marker, and function would require the highest. The CMD performed similarly to the DAS, requiring similar sample sizes, while the CPD would require 1.7 times greater sample size to distinguish treatment from placebo. The results were similar across DMARD and anti-TNF-alpha trials. CONCLUSION: Because of their demonstrated sensitivity to change, composite measures assessing RA outcomes in clinical trials should continue to include physician/assessor-derived core set measure assessments. PMID- 18412315 TI - LIGHT induces cell proliferation and inflammatory responses of rheumatoid arthritis synovial fibroblasts via lymphotoxin beta receptor. AB - OBJECTIVE: To investigate the effects of LIGHT (lymphotoxin-like, exhibits inducible expression and competes with herpes simplex virus glycoprotein D for herpes virus entry mediator, a receptor expressed by T lymphocytes) on the proliferation and gene expression of fibroblast-like synoviocytes (FLS) from patients with rheumatoid arthritis (RA). METHODS: We measured LIGHT levels in RA synovial fluids (SF) by ELISA, and compared them with those in osteoarthritis (OA) SF. Levels of LIGHT and its receptors in RA-FLS and synovium were assessed using real-time quantitative polymerase chain reaction (PCR). RA-FLS proliferation was examined by a bromodeoxyuridine assay. Expression of intercellular adhesion molecule-1 (ICAM-1) and several chemokines, such as interleukin 8 (IL-8), monocyte chemoattractant protein-1 (MCP-1), and macrophage inflammatory protein-1alpha (MIP-1alpha), was examined by real-time quantitative PCR, ELISA, and flow cytometry. The effects of LIGHT on nuclear factor-kappaB (NF kappaB) activation were investigated using immunofluorescence and Western blotting. RESULTS: LIGHT was upregulated in both SF and synovium of RA patients compared with OA patients. Herpes virus entry mediator (HVEM) and lymphotoxin beta receptor (LTbetaR), but not LIGHT, were detected in RA-FLS. LIGHT significantly promoted RA-FLS proliferation and induced expression of MCP-1, IL 8, MIP-1alpha, and ICAM-1 by RA-FLS. As well, LTbetaR small interfering RNA (siRNA), but not HVEM siRNA, inhibited these effects of LIGHT. LIGHT induced IkappaBa degradation and NF-kappaB translocation, and a NF-kappaB inhibitor suppressed the effects of LIGHT on RA-FLS. CONCLUSION: Our findings suggest that LIGHT signaling via LTbetaR plays an important role in the pathogenesis of RA by affecting key processes such as the proliferation and activation of RA-FLS. Regulation of LIGHT-LTbetaR signaling may represent a new therapeutic target for RA treatment. PMID- 18412316 TI - Design and microwave-assisted synthesis of novel macrocyclic peptides active at melanocortin receptors: discovery of potent and selective hMC5R receptor antagonists. AB - Differentiation of the physiological role of the melanocortin receptor 5 MC5R from that of other melanocortin receptors will require development of high affinity and selective antagonists. To date, a few synthetic antagonist ligands active at hMC5 receptor are available, but most do not have appreciable selectivity. With the aim to gain more potent and selective antagonists for the MC5R ligands, we have designed, synthesized, and pharmacologically characterized a series of alkylthioaryl-bridged macrocyclic peptide analogues derived from MT II and SHU9119. These 20-membered macrocycles were synthesized by a tandem combination using solid phase peptide synthesis and microwave-assisted reactions. Biological assays for binding affinities and adenylate cyclase activities for the hMC1R, hMC3R, hMC4R, and hMC5R showed that three analogues, compounds, 9, 4, and 7, are selective antagonists at the hMC5 receptor. In particular, compound 9(PG 20N) is a selective and competitive hMC5R antagonist, with IC 50 of 130 +/- 11 nM, and a pA 2 value of 8.3, and represents an important tool for further biological investigations of the hMC5R. Compounds 4 and 7 (PG14N, PG17N) show potent and selective allosteric inhibition at hMC5R with IC 50 values of 38 +/- 3 nM and 58 +/- 6 nM, respectively. Compound 9 will be used to further investigate and more clearly understand the physiological roles played by the MC5 receptor in humans and other animals. PMID- 18412317 TI - (3R,5S,E)-7-(4-(4-fluorophenyl)-6-isopropyl-2-(methyl(1-methyl-1h-1,2,4-triazol-5 yl)amino)pyrimidin-5-yl)-3,5-dihydroxyhept-6-enoic acid (BMS-644950): a rationally designed orally efficacious 3-hydroxy-3-methylglutaryl coenzyme-a reductase inhibitor with reduced myotoxicity potential. AB - 3-hydroxy-3-methylglutaryl coenzyme-A reductase (HMGR) inhibitors, more commonly known as statins, represent the gold standard in treating hypercholesterolemia. Although statins are regarded as generally safe, they are known to cause myopathy and, in rare cases, rhabdomyolysis. Statin-dependent effects on plasma lipids are mediated through the inhibition of HMGR in the hepatocyte, whereas evidence suggests that myotoxicity is due to inhibition of HMGR within the myocyte. Thus, an inhibitor with increased selectivity for hepatocytes could potentially result in an improved therapeutic window. Implementation of a strategy that focused on in vitro potency, compound polarity, cell selectivity, and oral absorption, followed by extensive efficacy and safety modeling in guinea pig and rat, resulted in the identification of compound 1b (BMS-644950). Using this discovery pathway, we compared 1b to other marketed statins to demonstrate its outstanding efficacy and safety profile. With the potential to generate an excellent therapeutic window, 1b was advanced into clinical development. PMID- 18412318 TI - Design and synthesis of conformationally constrained glucagon-like peptide-1 derivatives with increased plasma stability and prolonged in vivo activity. AB - A series of conformationally constrained derivatives of glucagon-like peptide-1 (GLP-1) were designed and evaluated. By use of [Gly (8)]GLP-1(7-37)-NH2 (2) peptide as a starting point, 17 cyclic derivatives possessing i to i + 4, i to i + 5, or i to i + 7 side chain to side chain lactam bridges from positions 18 to 30 were prepared. The effect of a helix-promoting alpha-amino-isobutyric acid (Aib) substitution at position 22 was also evaluated. The introduction of i to i + 4 glutamic acid-lysine lactam constraints in c[Glu (18)-Lys (22)][Gly (8)]GLP 1(7-37)-NH2 (6), c[Glu (22)-Lys (26)][Gly (8)]GLP-1(7-37)-NH2 (10), and c[Glu (23)-Lys (27)][Gly (8)]GLP-1(7-37)-NH2 (11) resulted in potent functional activity and receptor affinities comparable to native GLP-1. Selected GLP-1 peptides were chemoselectively PEGylated in order to prolong their in vivo activity. PEGylated peptides [Gly (8),Aib (22)]GLP-1(7-37)-Cys ((PEG))-Ala-NH2 (23) and c[Glu (22)-Lys (26)][Gly (8)]GLP-1(7-37)-Cys ((PEG))-Ser-Gly-NH2 (24) retained picomolar functional potency and avid receptor binding properties. Importantly, PEGylated GLP-1 peptide 23 exhibited sustained in vivo efficacy with respect to blood glucose reduction and decreased body weight for several days in nonhuman primates. PMID- 18412319 TI - Starburst triarylamine based dyes for efficient dye-sensitized solar cells. AB - We report here on the synthesis and photophysical/electrochemical properties of a series of novel starburst triarylamine-based organic dyes (S1, S2, S3, and S4) as well as their application in dye-sensitized nanocrystalline TiO2 solar cells (DSSCs). For the four designed dyes, the starburst triarylamine group and the cyanoacetic acid take the role of electron donor and electron acceptor, respectively. It was found that the introduction of starburst triarylamine group to form the D-D-pi-A configuration brought about superior performance over the simple D-pi-A configuration, in terms of bathochromically extended absorption spectra, enhanced molar extinction coefficients and better thermo-stability. Moreover, the HOMO and LUMO energy levels tuning can be conveniently accomplished by alternating the donor moiety, which was confirmed by electrochemical measurements and theoretical calculations. The DSSCs based on the dye S4 showed the best photovoltaic performance: a maximum monochromatic incident photon-to current conversion efficiency (IPCE) of 85%, a short-circuit photocurrent density (J(sc)) of 13.8 mA cm(-2), an open-circuit photovoltage (V(oc)) of 0.63 V, and a fill factor (ff) of 0.69, corresponding to an overall conversion efficiency of 6.02% under 100 mW cm(-2) irradiation. This work suggests that the dyes based on starburst triphenylamine donor are promising candidates for improvement of the performance of the DSSCs. PMID- 18412320 TI - A carbohydrate-based approach for the total synthesis of aculeatin D and 6-epi aculeatin D. AB - A concise approach for the total synthesis of aculeatin D and 6-epi-aculeatin D employing differentially protected anti, anti-1,3,5-triol alkyne prepared from alpha-D-glucoheptonic-gamma-lactone derivative is documented. Phenol protecting group manipulation for selective O-debenzylation during the hydrogenation of the diyne intermediate and one-pot phenolic oxidation with concomitant spiroketalization highlight the accomplished total synthesis. PMID- 18412321 TI - Effect of molecular and lattice structure on hydrogen transfer in molecular crystals of diamino-dinitroethylene and triamino-trinitrobenzene. AB - We have studied the intra- and intermolecular hydrogen transfer in a crystalline 1,1-diamino-2,2-dinitroethylene (DADNE) and 1,3,5-triamino-2,4,6-trinitrobenzene (TATB) by means of an embedded cluster method and density functional theory (DFT). We found that, even though both of these materials have similar amino- and nitro- functional groups and layered crystalline structures, there are important differences in the mechanisms of hydrogen transfer. In particular, our calculations suggest that the proton migration from an amino-group to a nitro group of the same molecule is a feasible process in TATB but not in DADNE. At the same time, we have found that no intermolecular hydrogen transfer occurs in either molecular crystal. These results imply that the activation of the decomposition reactions proceeds via different paths in these two materials. PMID- 18412322 TI - Implementation of a density functional theory-based method for the calculation of the hyperfine A-tensor in periodic systems with the use of numerical and Slater type atomic orbitals: application to paramagnetic defects. AB - The A-tensor parameterizes the "hyperfine" interaction of an "effective" electronic spin with the magnetic field due to the nuclear spin as monitored in an electron paramagnetic resonance (EPR) experiment. In this account, we describe an implementation for the calculation of the A-tensor in systems with translational invariance based on the Kohn-Sham form of density functional theory (KS DFT). The method is implemented in the periodic program BAND, where the Bloch states are expanded in the basis of numerical and Slater-type atomic orbitals (NAOs/STOs). This basis is well-suited for the accurate representation of the electron density near the nuclei, a prerequisite for the calculation of highly accurate hyperfine parameters. Our implementation does not rely on the frozen core approximation tacitly assumed in the pseudopotential schemes. The implementation is validated by performing calculations on the A-tensor for small atoms and molecules within the supercell approach as well as for paramagnetic defects in solids. In particular, we consider the A-tensor of "normal" and "anomalous" muonium defects in diamond and of the hydrogen cyanide anion radical HCN(-) in a KCl host crystal lattice. PMID- 18412324 TI - Synthesis and screening of mono- and di-aryl technetium and rhenium metallocarboranes. A new class of probes for the estrogen receptor. AB - A series of mono and diaryl rhenium(I)-carborane derivatives were prepared using microwave heating and screened for their affinity for two isoforms of the estrogen receptor (ER). The rhenacarborane derivative [(RR'C 2B9H9)Re(CO)3](-) (R = p-PhOH, R' = H), which was generated by taking advantage of a recently discovered cage isomerization process, and the neutral nitrosated analogue [(RR'C2B9H9)Re(CO)2(NO)] (R = p-PhOH, R' = H) showed the highest affinities of the compounds screened. As a result, the (99m)Tc analogue of one of the leads was produced in high yield (84%) and specific activity in a manner that is suitable for routine production in support of future preclinical and molecular imaging studies. PMID- 18412325 TI - Degradation of bidentate-coordinated platinum(II)-based DNA intercalators by reduced L-glutathione. AB - We have examined the interaction of [(5,6-dimethyl-1,10-phenanthroline)(1S,2S diaminocyclohexane)platinum(II)] (2+) (1, 56MESS), [(5-methyl-1,10 phenanthroline)(1S,2S-diaminocyclohexane)platinum(II)] (2+) (2, 5MESS), [(5,6 dimethyl-1,10-phenanthroline)(1R,2R-diaminocyclohexane)platinum(II)] (2+) (3, 56MERR), and [(5,6-dimethyl-1,10-phenanthroline)(ethylenediamine)platinum(II)] (2+) (4, 56MEEN) with reduced L-glutathione and L-methionine. Both thiols degrade all four complexes, mainly by displacing the ancillary ligand and forming a doubly bridged dinuclear complex. The degradation half-life of all the complexes with methionine is >7 days, indicating that these reactions are not biologically relevant. The rate of degradation by glutathione appears to be particularly important and shows an inverse correlation to cytotoxicity. The least active complex, 4 (t 1/2 glutathione: 20 h), degrades fastest, followed by 3 (31 h), 2 (40 h), and 1 (68 h). The major degradation product, [bis-mu-{reduced L glutathione}bis{5,6-dimethyl-1,10-phenanthroline}bis{platinum(II)}] (2+) (5, 56MEGL), displays no cytotoxicity and is excluded as the source of the anticancer activity. Once bound by glutathione, these metal complexes do not then form coordinate bonds with guanosine. Partial encapsulation of the complexes within cucurbit[n]urils is able to stop the degradation process. PMID- 18412326 TI - Novel potent and selective thrombin inhibitors based on a central 1,4-benzoxazin 3(4H)-one scaffold. AB - Novel thrombin inhibitors with the central 1,4-benzoxazine-3(4 H)-one scaffold, benzamidine P1 arginine side chain mimetic and various P3 moieties are described. 3-(Benzyl(2-(4-carbamimidoylbenzyl)-4-methyl-3-oxo-3,4-dihydro-2 H-1,4-benzoxazin 7-yl)amino)-3-oxopropanoic acid (7b), the most potent compound in the series, exhibited a K i of 2.6 nM in vitro for thrombin and high selectivity against trypsin and factor Xa. PMID- 18412327 TI - Structural origin of selectivity in class II-selective histone deacetylase inhibitors. AB - The development of class- and isoform-selective histone deacetylase (HDAC) inhibitors is highly desirable for the study of the complex interactions of these proteins central to transcription regulation as well as for the development of selective HDAC inhibitors as drugs in epigenetics. To provide a structural basis for the rational design of such inhibitors, a combined computational and experimental study of inhibition of three different histone deacetylase isoforms, HDAC1, -6, and -8, with three different hydroxamate inhibitors is reported. While SAHA was found to be unselective for the inhibition of class I and class II HDACs, the other inhibitors were found to be selective toward class II HDACs. Molecular dynamics simulations indicate that this selectivity is caused by both the overall shape of the protein surface leading to the active site and specific interactions of an aspartate residue in a polar loop and two phenylalanines and a methionine in a nonpolar loop. Monitoring the specific interactions as a function of the simulation time identifies a key sulfur-pi interaction. The implications of the structural motifs for the design of class II-selective HDAC inhibitors are discussed. PMID- 18412328 TI - Ranking targets in structure-based virtual screening of three-dimensional protein libraries: methods and problems. AB - Structure-based virtual screening is a promising tool to identify putative targets for a specific ligand. Instead of docking multiple ligands into a single protein cavity, a single ligand is docked in a collection of binding sites. In inverse screening, hits are in fact targets which have been prioritized within the pool of best ranked proteins. The target rate depends on specificity and promiscuity in protein-ligand interactions and, to a considerable extent, on the effectiveness of the scoring function, which still is the Achilles' heel of molecular docking. In the present retrospective study, virtual screening of the sc-PDB target library by GOLD docking was carried out for four compounds (biotin, 4-hydroxy-tamoxifen, 6-hydroxy-1,6-dihydropurine ribonucleoside, and methotrexate) of known sc-PDB targets and, several ranking protocols based on GOLD fitness score and topological molecular interaction fingerprint (IFP) comparison were evaluated. For the four investigated ligands, the fusion of GOLD fitness and two IFP scores allowed the recovery of most targets, including the rare proteins which are not readily suitable for statistical analysis, while significantly filtering out most false positive entries. The current survey suggests that selecting a small number of targets (<20) for experimental evaluation is achievable with a pure structure-based approach. PMID- 18412329 TI - Technique for generating three-dimensional alignments of multiple ligands from one-dimensional alignments. AB - We describe and demonstrate a method for the simultaneous, fully flexible alignment of multiple molecules with a common biological activity. The key aspect of the algorithm is that the alignment problem is first solved in a lower dimensional space, in this case using the one-dimensional representations of the molecules. The three-dimensional alignment is then guided by constraints derived from the one-dimensional alignment. We demonstrate using 10 hERG channel blockers, with a total of 72 rotatable bonds, that the one-dimensional alignment is able to effectively isolate key conserved pharmacophoric features and that these conserved features can effectively guide the three-dimensional alignment. Further using 10 estrogen receptor agonists and 5 estrogen receptor antagonists with publicly available cocrystal structures we show that the method is able to produce superpositions comparable to those derived from crystal structures. Finally, we demonstrate, using examples from peptidic CXCR3 agonists, that the method is able to generate reasonable binding hypotheses. PMID- 18412330 TI - Regioselectivity prediction of CYP1A2-mediated phase I metabolism. AB - A kinetic, reactivity-binding model has been proposed to predict the regioselectivity of substrates meditated by the CYP1A2 enzyme, which is responsible for the metabolism of planar-conjugated compounds such as caffeine. This model consists of a docking simulation for binding energy and a semiempirical molecular orbital calculation for activation energy. Possible binding modes of CYP1A2 substrates were first examined using automated docking based on the crystal structure of CYP1A2, and binding energy was calculated. Then, activation energies for CYP1A2-mediated metabolism reactions were calculated using the semiempirical molecular orbital calculation, AM1. Finally, the metabolic probability obtained from two energy terms, binding and activation energies, was used for predicting the most probable metabolic site. This model predicted 8 out of 12 substrates accurately as the primary preferred site among all possible metabolic sites, and the other four substrates were predicted into the secondary preferred site. This method can be applied for qualitative prediction of drug metabolism mediated by CYP1A2 and other CYP450 family enzymes, helping to develop drugs efficiently. PMID- 18412331 TI - Functional drift of sequence attributes in the FK506-binding proteins (FKBPs). AB - Diverse members of the FK506-binding proteins (FKBPs) group and their complexes with different macrocyclic ligands of fungal origins such as FK506, rapamycin, ascomycin, and their immunosuppressive and nonimmunosuppressive derivatives display a variety of cellular and biological activities. The functional relatedness of the FKBPs was estimated from the following attributes of their aligned sequences: 1 degrees conservation of the consensus sequence; 2 degrees sequence similarity; 3 degrees pI; 4 degrees hydrophobicity; 5 degrees amino acid hydrophobicity and bulkiness profiles. Analyses of the multiple sequence alignments and intramolecular interaction networks calculated from a series of structures of the FKBPs revealed some variations in the interaction clusters formed by the AA residues that are crucial for sustaining peptidylprolyl cis/trans isomerases (PPIases) activity and binding capacity of the FKBPs. Fine diversification of the sequences of the multiple paralogues and orthologues of the FKBPs encoded in different genomes alter the intramolecular interaction patterns of their structures and allowed them to gain some selectivity in binding to diverse targets (functional drift). PMID- 18412332 TI - Transfer of copper between bis(thiosemicarbazone) ligands and intracellular copper-binding proteins. insights into mechanisms of copper uptake and hypoxia selectivity. AB - Bis(thiosemicarbazonato) complexes Cu(II)(Btsc) have attracted interest as promising metallodrugs and, in particular, as copper radiopharmaceuticals. Prototypes Cu(Atsm) and Cu(Gtsm) are membrane-permeable, but their metabolisms in cells are distinctly different: copper that is delivered by Cu(Gtsm) is trapped nonselectively in all cells, whereas copper that is delivered by Cu(Atsm) is retained selectively in hypoxic cells but is "washed out" readily in normal cells. We have studied copper-transfer reactions of these two complexes under various conditions, aiming to model their cellular chemistry. In Me2SO, both complexes exhibited reversible one-electron-reduction processes with Cu(Atsm) being more difficult to reduce than Cu(Gtsm) (E(1/2)'=-0.60 and -0.44 V, respectively, vs AgCl/Ag). Upon introduction of an aqueous buffer into Me2SO, the electrochemical reduction remained chemically reversible for Cu(Atsm) but became irreversible for Cu(Gtsm). However, the estimated difference in their reduction potentials did not change. Chromophoric ligand anions bicinchonate (Bca) and bathocuproine disulfonate (Bcs) were used as Cu(I) indicators to trace the destinations of copper in the reactions and to mimic cellular Cu(I)-binding components ("sinks"). While both BtscH2 ligands have high affinities for Cu(I) (KD in the picomolar range), they cannot compete with Cu(I) sinks such as the copper-binding proteins Atx1 and Ctr1c (or a mimic such as Bcs). In the presence of these proteins, reduction of Cu(II)(Btsc) leads to irreversible transfer of copper to the protein ligands. Endogenous reductants ascorbate and glutathione can reduce Cu(II)(Gtsm) in the presence of such protein ligands but cannot reduce Cu(II)(Atsm). These properties establish a strong correlation between the contrasting cellular retention properties of these complexes and their different reduction potentials. The endogenous reductants in normal cells appear to be able to reduce Cu(II)(Gtsm) but not Cu(II)(Atsm), allowing the latter to be washed out. The more reducing environment of hypoxic cells leads to reduction of Cu(II)(Atsm) and retention of its copper. PMID- 18412333 TI - Intrinsic carrier doping in antiferromagnetically interacted supramolecular copper complexes with (pyrazino)tetrathiafulvalene (pyra-TTF) as the ligand, [CuIICl2(pyra-TTF)] and (pyra-TTF)2[CuI3Cl4(pyra-TTF)]. AB - New supramolecular copper complexes with pyrazinotetrathiafulvalene (pyra-TTF) as the ligand, [Cu(II)Cl2(pyra-TTF)] (1) and (pyra-TTF) 2[Cu(I)3Cl4(pyra-TTF)] (2), have been synthesized by the diffusion method. Complex 1 is a black block crystal with a three-dimensional (3-D) supramolecular network; the linear chain [ Cu(II)Cl2-(pyra-TTF)-] n extends along the b axis, where the coordinated pyra-TTF donors are stacked in a head-to-tail and ring-over-bond configuration to construct two-dimensional (2-D) sheets, and between the sheets, there are C...Cl( ) or H...Cl(-) contacts. Even though the electron spin resonance (ESR) measurement reveals the nearly Cu(II) state, complex 1 is a semiconductor with sigmaRT=1.0 x 10(-4) S cm(-1) and Ea=0.33 eV. The high-frequency conductivity measurement also confirmed the intrinsic slight carrier doping from Cu(II) to the pyra-TTF donor. This slight doping enhances not only the real and imaginary dielectric constants but also the antiferromagnetic interaction between Cu(II) spins following the 2-D Heisenberg model with 2J=-20 K. In contrast, complex 2 is a very thin black needle. This needle crystal has two crystallographically independent pyra-TTF molecules, which are coordinated and noncoordinated donors. The coordinated donors composed a supramolecular chain [Cu(I)3Cl4(pyra-TTF)(0)]n , whereas the noncoordinated donors formed conducting alpha''-type pyra-TTF(+0.5) sheets. This complex is semiconducting with sigmaRT=0.1 S cm(-1) and Ea=0.15 eV. Both complexes 1 and 2 demonstrate that the pyra-TTF molecule works not only as an oxidized donor by Cu(II) to construct conducting sheets but also as a ligand coordinated to a Cu cation to form supramolecuar chains. PMID- 18412334 TI - Binuclear copper(II) complexes with N4O3 coordinating heptadentate ligand: synthesis, structure, magnetic properties, density-functional theory study, and catecholase activity. AB - The N4O3 coordinating heptadentate ligand afforded binuclear complex [Cu 2(H 2L)(mu-OH)](ClO4)2 (1) and [Cu2(L)(H2O)2]PF6 (2). In complex 1, two copper ions are held together by mu-phenoxo and mu-hydroxo bridges, whereas in complex 2, the copper centers are connected only by a mu-phenoxo bridge. In 1, both the Cu(II) centers have square pyramidal geometry (tau=0.01-0.205), whereas in the case of 2, one Cu(II) center has square pyramidal (tau=0.2517) and other one has square based pyramidal distorted trigonal bipyramidal (tau=0.54) geometry. Complexes 1 and 2 show an strong intramolecular and very weak antiferromagnetic interaction, respectively. Density-functional theory calculations were performed to establish the magneto structural correlation between the two paramagnetic copper(II) centers. Both of the complexes display a couple of one-electron reductive responses near -0.80 and -1.10 V. The complexes show significant catalytic activity at pH 8.5 on the oxidation of 3,5-di- tert-butylcatechol (3,5-DTBC) to 3,5-di- tert-butylquinone (3,5-DTBQ), and the activity measured in terms of kcat=29-37 h(-1). PMID- 18412335 TI - ZnO twin-cones: synthesis, photoluminescence, and catalytic decomposition of ammonium perchlorate. AB - ZnO twin-cones, a new member to the ZnO family, were prepared directly by a solvothermal method using a mixed solution of zinc nitrate and ethanol. The reaction and growth mechanisms of ZnO twin-cones were investigated by X-ray diffraction, UV-visible spectra, infrared and ion trap mass spectra, and transmission electron microscopy. All as-prepared ZnO cones consisted of tiny single crystals with lengths of several micrometers. With prolonging of the reaction time from 1.5 h to 7 days, the twin-cone shape did not change at all, while the lattice parameters increased slightly and the emission peak of photoluminescence shifted from the green region to the near orange region. ZnO twin-cones are also explored as an additive to promote the thermal decomposition of ammonium perchlorate. The variations of photoluminescence spectra and catalytic roles in ammonium perchlorate decomposition were discussed in terms of the defect structure of ZnO twin-cones. PMID- 18412336 TI - Synthesis and characterization of three homoleptic alkoxides of uranium: [Li(THF)]2[UIV(OtBu)6], [Li(Et2O)][UV(OtBu)6], and UVI(OtBu)6. AB - Addition of 6 equiv of LiOtBu to a THF/Et2O solution of UCl4 at -25 degrees C generates [Li(THF)]2[U(OtBu)6] (1) in 61% yield. 1 is soluble in polar organic solvents and is stable for several days in THF. However, 1 slowly decomposes in benzene or hexanes, forming the dinuclear uranium(IV) species [Li(THF)][U2(OtBu)9] (2) as one of the decomposition products. Alternatively, 2 can be directly prepared in moderate yield by the addition of 4.5 equiv LiOtBu to UCl4 in hexanes/THF at room temperature. The decomposition of 1 has been studied by 1H and 7Li{1H} NMR spectroscopies to elucidate the nature of this transformation. Oxidation of 1 occurs readily in the presence of 0.5 or 1 equiv of I2 to give [Li(Et2O)][U(OtBu)6] (3) and U(OtBu)6 (4), respectively, in good yields. Alternately, 3 can be generated by comproportionation of 1 and 4. 1-4 have been fully characterized, including analysis by X-ray crystallography. In the solid-state these complexes possess large U-O-Cq bond angles, suggestive of a significant U-O pi interaction. In addition, we have studied the redox properties of 4 by cyclic voltammetry. PMID- 18412337 TI - Reactivity of a phosphinito-bridged PtI-PtI complex with nucleophiles: substitution versus addition. AB - As a result of the strong electrophilic character of the Pt bound to O, the phosphinito-bridged PtI complex [(PHCy2)Pt(micro-PCy2){kappa2P,O-micro P(O)Cy2}Pt(PHCy2)](Pt-Pt) (1) undergoes attack at the O-bound Pt atom by molecules such as di- and tricyclohexylphosphane, dicyclohexylphosphane oxide, and dicyclohexylphosphane sulfide. Thus, reaction of 1 with PHCy2 gives the symmetric PtI dimer [(PHCy2)Pt(micro-PCy2)]2(Pt-Pt) (2), while the hydrido bridged complex syn-[(PHCy2){kappaP-P(O)Cy2}Pt(micro-PCy2)(micro H)Pt(PHCy2){kappaP-P(O)Cy2}](Pt-Pt) (4) is obtained from reaction of 1 with P(O)HCy2; the thiophosphinito complex [(PHCy2)Pt(micro-PCy2){kappa2P,S-micro P(S)Cy2}Pt(PHCy2)](Pt-Pt) (8) forms selectively in reaction of 1 with P(S)HCy2. For comparison, the reaction with PCy3 results only in ligand substitution, affording [(PCy3)Pt(micro-PCy2){kappa2P,O-micro-P(O)Cy2}Pt(PHCy2)](Pt-Pt) (5). DFT studies confirmed the remarkable electrophilicity of the oxygen-bound Pt and shed light on the nature of the metal-metal bond in Pt dimers. PMID- 18412338 TI - Ambipolar, high performance, acene-based organic thin film transistors. AB - We present a high performance, ambipolar organic field-effect transistor composed of a single material. Ambipolar molecules are rare, and they can enable low-power complementary-like circuits. This low band gap, asymmetric linear acene contains electron-withdrawing fluorine atoms, which lower the molecular orbital energies, allowing the injection of electrons. While hole and electron mobilities of up to 0.071 and 0.37 cm2/V.s, respectively, are reported on devices measured in nitrogen, hole mobilities of up to 0.12 cm2/V.s were found in ambient, with electron transport quenched. These devices were fabricated on octadecyltrimethoxysilane-treated surfaces at a substrate temperature of 60 degrees C. PMID- 18412339 TI - Measurement of the binding of cholera toxin to GM1 gangliosides on solid supported lipid bilayer vesicles and inhibition by europium (III) chloride. AB - In this paper the immobilization of small unilamellar DMPC/GM1 lipid vesicles containing a water-soluble bodipy dye is described. The binding of the complete alphabeta toxin expressed by Vibrio cholerae to the attached vesicles was measured using Surface Plasmon Resonance (SPR) and a value of the dissociation constant K d obtained. Further measurements showed that the interaction of both the alphabeta-toxin and the beta-subunit alone resulted in the permeation of the lipid membrane, with release of a fluorophore contained within the vesicle being measured by combined SPR and Surface Plasmon enhanced Fluorescence Spectroscopy (SPFS). The leakage of dye through the membrane, measured by following the change in fluorescence, was fitted to a simple diffusion model. Finally, SPFS measurements of the effect of europium(III) chloride (EuCl 3) showed that cholera toxin binding and subsequent membrane permeation could be blocked by 1 micromol dm (-3) europium chloride. In view of the low oral toxicity of europium chloride, we speculate on the potential pharmaceutical applications of this molecule in the treatment of cholera infection. PMID- 18412340 TI - Effect of a tridentate ligand on the structure, electronic structure, and reactivity of the copper(I) nitrite complex: role of the conserved three histidine ligand environment of the type-2 copper site in copper-containing nitrite reductases. AB - It is postulated that the copper(I) nitrite complex is a key reaction intermediate of copper containing nitrite reductases (Cu-NiRs), which catalyze the reduction of nitrite to nitric oxide (NO) gas in bacterial denitrification. To investigate the structure-function relationship of Cu-NiR, we prepared five new copper(I) nitrite complexes with sterically hindered tris(4 imidazolyl)carbinols [Et-TIC = tris(1-methyl-2-ethyl-4-imidazolyl)carbinol and iPr-TIC = tris(1-methyl-2-isopropyl-4-imidazolyl)carbinol] or tris(1 pyrazolyl)methanes [Me-TPM = tris(3,5-dimethyl-1-pyrazolyl)methane; Et-TPM = tris(3,5-diethyl-1-pyrazolyl)methane; and iPr-TPM = tris(3,5-diisopropyl-1 pyrazolyl)methane]. The X-ray crystal structures of all of these copper(I) nitrite complexes were mononuclear eta(1)-N-bound nitrite complexes with a distorted tetrahedral geometry. The electronic structures of the complexes were investigated by absorption, magnetic circular dichroism (MCD), NMR, and vibrational spectroscopy. All of these complexes are good functional models of Cu NiR that form NO and copper(II) acetate complexes well from reactions with acetic acid under anaerobic conditions. A comparison of the reactivity of these complexes, including previously reported (iPr-TACN)Cu(NO2) [iPr-TACN = 1,4,7 triisopropyl-1,4,7-triazacyclononane], clearly shows the drastic effects of the tridentate ligand on Cu-NiR activity. The copper(I) nitrite complex with the Et TIC ligand, which is similar to the highly conserved three-histidine ((His)3) ligand environment in the catalytic site of Cu-NiR, had the highest Cu-NiR activity. This result suggests that the (His)3 ligand environment is essential for acceleration of the Cu-NiR reaction. The highest Cu-NiR activity for the Et TIC complex can be explained by the structural and spectroscopic characterizations and the molecular orbital calculations presented in this paper. Based on these results, the functional role of the (His)3 ligand environment in Cu-NiR is discussed. PMID- 18412342 TI - Functionalized methanofullerenes used as n-type materials in bulk-heterojunction polymer solar cells and in field-effect transistors. AB - The synthesis of two well-solubilized [60]methanofullerene derivatives ( p- EHO PCBM and p- EHO-PCBA) is presented for usage in organic solar cells and in field effect transistors. The para position of the PCBM's phenyl ring was substituted with a branched alkoxy side chain, which contributes to higher solubility, facilitating synthesis, purification, and processing. We find a small change of the open-circuit voltage ( V oc) as a slight improvement in performance upon application in P3HT/[60]methanofullerene bulk-heterojunction-photovoltaic cells, when compared to PCBM, because of the electron donation of the alkoxy group. In the case of the devices with a TiO x layer, the best power conversion efficiencies (PCE, eta e) is observed in a layered structure of P3HT/ p- EHO PCBA/TiO x (eta e = 2.6%), which slightly exceeds that of P3HT/PCBM/TiO x (eta e = 2.3%) under conditions reported here. This can be attributed, in part, to the carboxylic acid group in p- EHO-PCBA that leads to an effective interface interaction between the active layer and TiO x phase. In addition, n-channel organic field-effect transistor (OFET) devices were fabricated with thin films of p- EHO-PCBM and p- EHO-PCBA, respectively cast from solution on SiO 2/Si substrates. The values of field-effect mobility (mu) for p- EHO-PCBM and p- EHO PCBA are 1 x 10 (-2) and 1.6 x 10 (-3) cm (2)/V.s, respectively. The results in this paper demonstrate the effects of a carboxylic acid group and an electron donating substituent in [60]methanofullerenes as n-type materials with respect to organic solar cells and OFET applications. PMID- 18412341 TI - Carbon-deuterium bonds as probes of dihydrofolate reductase. AB - Much effort has been directed toward understanding the contributions of electrostatics and dynamics to protein function and especially to enzyme catalysis. Unfortunately, these studies have been limited by the absence of direct experimental probes. We have been developing the use of carbon-deuterium bonds as probes of proteins and now report the application of the technique to the enzyme dihydrofolate reductase, which catalyzes a hydride transfer and has served as a paradigm for biological catalysis. We observe that the stretching absorption frequency of (methyl- d 3) methionine carbon-deuterium bonds shows an approximately linear dependence on solvent dielectric. Solvent and computational studies support the empirical interpretation of the stretching frequency in terms of local polarity. To begin to explore the use of this technique to study enzyme function and mechanism, we report a preliminary analysis of (methyl- d 3) methionine residues within dihydrofolate reductase. Specifically, we characterize the IR absorptions at Met16 and Met20, within the catalytically important Met20 loop, and Met42, which is located within the hydrophobic core of the enzyme. The results confirm the sensitivity of the carbon-deuterium bonds to their local protein environment, demonstrate that dihydrofolate reductase is electrostatically and dynamically heterogeneous, and lay the foundation for the direct characterization protein electrostatics and dynamics and, potentially, their contribution to catalysis. PMID- 18412343 TI - Synthesis, resolution, and VCD analysis of an enantiopure diazaoxatricornan derivative. AB - Using simple organic synthetic transformations, a novel diazaoxatricornan derivative, the 12 c-methyl-12-phenyl-8-propyl-12,12 c-dihydro-8 H-4-oxa-8,12 diazadibenzo[ cd, mn]pyrene ( 6a), was prepared. This novel chiral cup-shaped molecule was isolated in racemic form and in excellent yield after the addition of methyl lithium to the BF 4 salt of a novel unsymmetrical diazaoxatriangulenium cation. Compound 6a was found to be stable under classical laboratory conditions something not obvious considering the extreme stability of the carbenium ion precursor, the electron-rich nature of the core, and the strain induced by the pyramidalization of the central carbon. The enantiomers were readily separated by chiral stationary phase chromatography, and the absolute configuration of (-)-( S)- 6a was determined by a comparison of the experimental and theoretical vibrational circular dichroism (VCD) spectra. This isolation of (-)-( S)- 6a and (+)-( R)- 6a constitutes thus the first report of a nonracemic closed-capped chiral bowl molecule for which the chirality is due to the intrinsic dissymmetry of the central core of the structure only. PMID- 18412344 TI - The total synthesis of roquefortine C and a rationale for the thermodynamic stability of isoroquefortine C over roquefortine C. AB - The first total synthesis of roquefortine C is achieved by implementation of a novel elimination strategy to construct the thermodynamically unstable E dehydrohistidine moiety. Molecular modeling studies are presented which explain the instability of the roquefortine C structure compared to that of isoroquefortine C. PMID- 18412345 TI - Hydrogen bonding contributes to the selectivity of nucleotide incorporation opposite an oxidized abasic lesion. AB - The ability of DNA polymerases to maintain the integrity of the genome even after it has been structurally altered is vital. There is considerable interest in determining the structural properties of the DNA template that polymerases recognize when determining which nucleotide to add to a nascent strand. Mechanistic, synthetic, and structural chemistries have been used to study how DNA polymerase activity is affected by size, shape, pi-stacking, and hydrogen bonds of the template molecules. Herein, we probe the structural aspects of abasic lesions that result in their distinct coding potential in Escherichia coli despite lacking a Watson-Crick base. In particular, we investigate why bypass of 2-deoxyribonolactone (L) results in significant amounts of dG incorporation opposite the lesion, whereas other abasic lesions (e.g., AP) adhere to the "A rule". Experiments using synthetic analogues reveal that DNA polymerase V bypasses L and increased levels of dG incorporation result from a hydrogen bonding interaction between the carbonyl oxygen and dG. These results show that a DNA polymerase utilizes hydrogen bonding as one structural parameter when decoding an abasic lesion. PMID- 18412347 TI - Chemical reactivity of D3h C78 (metallo)fullerene: regioselectivity changes induced by Sc3N encapsulation. AB - We report here for the first time a full comparison of the exohedral reactivity of a given fullerene and its parent trinitride template endohedral metallofullerene. In particular, we study the thermodynamics and kinetics for the Diels-Alder [4 + 2] cycloaddition between 1,3-butadiene and free D3h'-C78 fullerene and between butadiene and the corresponding endohedral D3h-Sc3N@C78 derivative. The reaction is studied for all nonequivalent bonds, in both the free and the endohedral fullerenes, at the BP86/TZP//BP86/DZP level. The change in exohedral reactivity and regioselectivity when a metal cluster is encapsulated inside the cage is profound. Consequently, the Diels-Alder reaction over the free fullerene and the endohedral derivative leads to totally different cycloadducts. This is caused by the metal nitride situated inside the fullerene cage that reduces the reactivity of the free fullerene and favors the reaction over different bonds. PMID- 18412346 TI - Structure and dynamics of the Abeta(21-30) peptide from the interplay of NMR experiments and molecular simulations. AB - We combine molecular dynamics simulations and new high-field NMR experiments to describe the solution structure of the Abeta(21-30) peptide fragment that may be relevant for understanding structural mechanisms related to Alzheimer's disease. By using two different empirical force-field combinations, we provide predictions of the three-bond scalar coupling constants ((3)J(H(N)H(alpha))), chemical-shift values, (13)C relaxation parameters, and rotating-frame nuclear Overhauser effect spectroscopy (ROESY) crosspeaks that can then be compared directly to the same observables measured in the corresponding NMR experiment of Abeta(21-30). We find robust prediction of the (13)C relaxation parameters and medium-range ROESY crosspeaks by using new generation TIP4P-Ew water and Amber ff99SB protein force fields, in which the NMR validates that the simulation yields both a structurally and dynamically correct ensemble over the entire Abeta(21-30) peptide. Analysis of the simulated ensemble shows that all medium-range ROE restraints are not satisfied simultaneously and demonstrates the structural diversity of the Abeta(21-30) conformations more completely than when determined from the experimental medium-range ROE restraints alone. We find that the structural ensemble of the Abeta(21-30) peptide involves a majority population (approximately 60%) of unstructured conformers, lacking any secondary structure or persistent hydrogen-bonding networks. However, the remaining minority population contains a substantial percentage of conformers with a beta-turn centered at Val24 and Gly25, as well as evidence of the Asp23 to Lys28 salt bridge important to the fibril structure. This study sets the stage for robust theoretical work on Abeta(1-40) and Abeta(1-42), for which collection of detailed NMR data on the monomer will be more challenging because of aggregation and fibril formation on experimental timescales at physiological conditions. In addition, we believe that the interplay of modern molecular simulation and high quality NMR experiments has reached a fruitful stage for characterizing structural ensembles of disordered peptides and proteins in general. PMID- 18412348 TI - Nickel-catalyzed decarbonylative addition of phthalimides to alkynes. AB - An intermolecular nickel-catalyzed addition reaction has been developed where N arylphthalimides react with alkynes to afford substituted isoquinolones. A mechanistic rationale is proposed, implying nucleophilic attack of Ni(0) to an amide as the primary step of the catalytic cycle. PMID- 18412350 TI - Palladium vacataporphyrin reveals conformational rearrangements involving Huckel and Mobius macrocyclic topologies. AB - 5,10,15,20-Tetraaryl-21-vacataporphyrin (butadieneporphyrin, an annulene porphyrin hybrid) which contains a vacant space instead of heteroatomic bridge acts as a ligand toward palladium(II). The metal ion of square-planar coordination geometry is firmly held via three pyrrolic nitrogen atoms where the fourth coordination place is occupied by a monodentate ligand or by an annulene part of vacataporphyrin. The macrocycle reveals the unique structural flexibility triggered by coordination of palladium. The structural rearrangements engage the C(20)C(1)C(2)C(3)C(4)C(5) annulene fragment which serves as a linker between two pyrrolic rings of vacataporphyrin albeit the significant ruffling of the tripyrrolic block is also of importance. Two fundamental modes of interactions between the palladium ion and annulene moiety have been recognized. The first one resembles an eta(2)-type interaction and involves the C(2)C(3) unit of the butadiene part. Alternatively the profound conformational adjustments allowed an in-plane coordination through the deprotonated trigonally hybridized C(2) center of butadiene. The coordinated vacataporphyrin acquires Huckel or extremely rare Mobius topologies readily reflected by spectroscopic properties. The palladium vacataporphyrin complexes reveal Huckel aromaticity or Mobius antiaromaticity of [18]annulene applying the butadiene fragment of vacataporphyrin as a topology selector. The properties of specific conformers were determined using (1)H NMR and density functional theory calculations. PMID- 18412351 TI - An Efficient, one-pot synthesis of isomeric ellipticine derivatives through intramolecular imino-Diels-Alder reaction. AB - New analogues of isomeric ellipticine derivatives fused with biologically important pyrroloindole or chromene moiety have been synthesized by utilizing an intramolecular imino Diels-Alder reaction in a single step. PMID- 18412349 TI - HIV-1 protease inhibitors from inverse design in the substrate envelope exhibit subnanomolar binding to drug-resistant variants. AB - The acquisition of drug-resistant mutations by infectious pathogens remains a pressing health concern, and the development of strategies to combat this threat is a priority. Here we have applied a general strategy, inverse design using the substrate envelope, to develop inhibitors of HIV-1 protease. Structure-based computation was used to design inhibitors predicted to stay within a consensus substrate volume in the binding site. Two rounds of design, synthesis, experimental testing, and structural analysis were carried out, resulting in a total of 51 compounds. Improvements in design methodology led to a roughly 1000 fold affinity enhancement to a wild-type protease for the best binders, from a Ki of 30-50 nM in round one to below 100 pM in round two. Crystal structures of a subset of complexes revealed a binding mode similar to each design that respected the substrate envelope in nearly all cases. All four best binders from round one exhibited broad specificity against a clinically relevant panel of drug-resistant HIV-1 protease variants, losing no more than 6-13-fold affinity relative to wild type. Testing a subset of second-round compounds against the panel of resistant variants revealed three classes of inhibitors: robust binders (maximum affinity loss of 14-16-fold), moderate binders (35-80-fold), and susceptible binders (greater than 100-fold). Although for especially high-affinity inhibitors additional factors may also be important, overall, these results suggest that designing inhibitors using the substrate envelope may be a useful strategy in the development of therapeutics with low susceptibility to resistance. PMID- 18412352 TI - Synthesis of maremycins A and D1 via cycloaddition of a nitrone with (E)-3 ethylidene-1-methylindolin-2-one. AB - A concise synthesis of maremycins A and D1 has been accomplished via cycloaddition of a chiral cyclic nitrone with ( E)-3-ethylidene-1-methylindolin-2 one as a key step. This synthesis clarifies the stereochemistry of the maremycins and is suitable for large-scale synthesis for biological screening. PMID- 18412353 TI - First total synthesis of cassiarin A, a naturally occurring potent antiplasmodial alkaloid. AB - The first total synthesis of cassiarin A, an antiplasmodial alkaloid isolated from Cassia siamea, was achieved via sequential alkynylation of arenes with Sonogashira coupling and 6- endo-dig-cyclization of phenolic oxygens to the resulting alkynes. PMID- 18412354 TI - A facile preparation of imidazolinium chlorides. AB - A process for the preparation of symmetric and unsymmetric imidazolinium chlorides that involves reaction of a formamidine with dichloroethane and a base (a) is described. This method makes it possible to obtain numerous imidazolinium chlorides under solvent-free reaction conditions and in excellent yields with purification by simple filtration. Alternatively, symmetric imidazolinium chlorides can be prepared directly in moderate yields from substituted anilines by utilizing half of the formamidine intermediate as sacrificial base (b). PMID- 18412355 TI - Regio- and stereoselective synthesis of alkyl allylic ethers via gold(I) catalyzed intermolecular hydroalkoxylation of allenes with alcohols. AB - Reaction of 1-phenyl-1,2-butadiene with 2-phenyl-1-ethanol catalyzed by a 1:1 mixture of a gold(I) N-heterocyclic carbene complex and AgOTf at room temperature for 1 h led to isolation of (E)-(3-phenethoxy-1-butenyl)benzene in 96% yield as a single regio- and stereoisomer. Gold(I)-catalyzed intermolecular allene hydroalkoxylation was effective for monosubsituted, 1,1- and 1,3-disubstituted, trisubstituted, and tetrasubstituted allenes and for a range of primary and secondary alcohols, methanol, phenol, and propionic acid. PMID- 18412356 TI - Increasing the persistency of stable free-radicals: synthesis and characterization of a nitroxide based [1]rotaxane. AB - A persistent paramagnetic [1]rotaxane based on beta-cyclodextrin showing an increased persistency under reductive conditions has been prepared and characterized for the first time. PMID- 18412357 TI - Phenolic composition in grape (Vitis vinifera L. cv. Malbec) ripened with different solar UV-B radiation levels by capillary zone electrophoresis. AB - The responses of Vitis vinifera L. cv. Malbec to different solar ultraviolet-B radiation (UV-B) levels were assessed in two contrasting situations, under sunlight with full UV-B (+UV-B) and filtered UV-B (-UV-B), in three different locations at 500, 1000, and 1500 m above sea level (asl). To evaluate the effects of radiation, a simple, accurate, and rapid method for the separation and simultaneous determination of representative phenolic compounds in grape berry skins by capillary zone electrophoresis was developed. Separation was carried out in less than 20 min with 20 mM sodium tetraborate buffer containing 30% methanol, pH 9.00. The procedure is fast and reliable, and extracted grape berry skins can be directly analyzed without prior sample cleanup procedure. Berry skins from the +UV-B treatment at 1500 m asl showed the highest levels of total polyphenols anthocyanins, and resveratrol, compared with the -UV-B treatment at this altitude. PMID- 18412358 TI - Quantification of zearalenone in various solid agroenvironmental samples using D6 zearalenone as the internal standard. AB - Because of its pronounced estrogenicity, zearalenone may be of concern not only in the aqueous but also in the terrestrial environment. Therefore, we developed several analytical methods to quantify zearalenone in different solid matrices of agroenvironmental relevance (i.e., plant organs, soil, manure, and sewage sludge). The use of D(6)-zearalenone as the internal standard (IS) was essential to render the analytical method largely matrix-independent because it compensated for target analyte losses during extract treatment and ion suppression during ionization. Soil and sewage sludge samples were extracted with Soxhlet, whereas plant material and manure samples were extracted by liquid solvent extraction at room temperature. Absolute recoveries for zearalenone were 70-104% for plant materials, 105% for soil, 76% for manure, and 30% for sewage sludge. Relative recoveries ranged from 86 to 113% for all matrices, indicating that the IS was capable to largely compensate for losses during analysis. Ion suppression, between 8 and 74%, was in all cases compensated by the IS but influenced the method quantification levels. These were 3.2-26.2 ng/g(dryweightdw) for plant materials, 0.7 ng/g(dw) for soil, 12.3 ng/g(dw) for manure, and 6.8 ng/g(dw) for sewage sludge. Plant material concentrations varied from 86 ng/g(dw) to more than 16.7 microg/g(dw), depending on the organ and crop. Soil concentrations were between not detectable and 7.5 ng/g(dw), depending on the sampling depth. Zearalenone could be quantified in all manure samples in concentrations between 8 and 333 ng/g(dw). Except for two of the 85 investigated sewage sludge samples, zearalenone concentrations were below quantification limit. PMID- 18412359 TI - Synthesis of oligosaccharides derived from lactulose and pectinex ultra SP-L. AB - The beta-galactosidase activity of the commercial enzymatic preparation Pectinex Ultra SP-L derived from Aspergillus aculeatus has been used to hydrolyze and transgalactosylate the prebiotic carbohydrate lactulose. During this reaction, new oligosaccharides derived from lactulose have been detected by high performance anion-exchange chromatography with pulsed amperometric detection (HPAEC-PAD). The presence of the trisaccharide 6'-galactosyl-lactulose, the major compound formed, has been confirmed by NMR. In addition, disaccharides and other oligosaccharides with higher retention times have been also detected. The effect of transgalactosylation conditions such as time, temperature, pH, and initial lactulose and enzyme concentrations, as well as product inhibition on oligosaccharide synthesis, has been studied. The optimal conditions for the formation of tri and higher oligosaccharides were 60 degrees C, pH 6.5, 450 g/L lactulose, 16 units/mL of enzyme, and 7 h of reaction. Selective formation of disaccharides was achieved under the same conditions with the exception of pH (4.5). The present work provides additional knowledge on the synthesis of new oligosaccharides with potential prebiotic properties. PMID- 18412360 TI - Characterization of steam volatiles in the essential oil of black currant buds and the antioxidant properties of different bud extracts. AB - Currently essential oil from black currant ( Ribes nigrum L.) buds is mainly used as a valuable perfumery ingredient. This study reports more comprehensive characterization of dormant buds of various black currant ( Ribes nigrum L.) cultivars which are grown in Northern European countries. Essential oils were isolated from the buds by hydrodistillation and analyzed by gas chromatography mass spectrometry (GC-MS), GC-flame ionization detection (GC-FID), and GC- olfactometry (GC-O). The most abundant compounds in black currant bud essential oil were sabinene, delta-3-carene, and terpinolene. The most frequent descriptors of the essential oil components assessed by GC-O were "woody", "terpene", "fruity", "sweet", "citrus", "herbaceous", "pine", "green", "oily", "herbal", and "musty". The residues obtained after hydrodistillation were separated into liquid and solid fractions. The solid fraction was dried and extracted with acetone (AE), while the liquid fraction (water extract) was divided in two parts, one of which was spray-dried (SDWF extracts) and the other freeze-dried (FDWF extract). In addition, a portion of whole frozen buds was extracted with methanol (ME). The radical scavenging capacity (RSC) of black currant bud extracts varied in a wide range; in the DPPH * reaction system FDWF at the applied concentration scavenged 43-79%; SDWF, 54-80%; AE, 16-36%; ME, 42-60% of radicals; while in the ABTS *+ reaction system the RSC was 39-72, 38-53, 1-5, and 30-49%, respectively. The total amount of phenolic compounds expressed in gallic acid equivalents in FDWF varied in the range of 132-192 mg/g; in SDWF, 140-209 mg/g; in AE, 49-107 mg/g; and in ME extracts, 111-180 mg/g. PMID- 18412361 TI - Berry extracts exert different antiproliferative effects against cervical and colon cancer cells grown in vitro. AB - Polyphenol-rich berry extracts were screened for their antiproliferative effectiveness using human cervical cancer (HeLa) cells grown in microtiter plates. Rowan berry, raspberry, lingonberry, cloudberry, arctic bramble, and strawberry extracts were effective but blueberry, sea buckthorn, and pomegranate extracts were considerably less effective. The most effective extracts (strawberry > arctic bramble > cloudberry > lingonberry) gave EC 50 values in the range of 25-40 microg/(mL of phenols). These extracts were also effective against human colon cancer (CaCo-2) cells, which were generally more sensitive at low concentrations but conversely less sensitive at higher concentrations. The strawberry, cloudberry, arctic bramble, and the raspberry extracts share common polyphenol constituents, especially the ellagitannins, which have been shown to be effective antiproliferative agents. However, the components underlying the effectiveness of the lingonberry extracts are not known. The lingonberry extracts were fractionated into anthocyanin-rich and tannin-rich fractions by chromatography on Sephadex LH-20. The anthocyanin-rich fraction was considerably less effective than the original extract, whereas the antiproliferative activity was retained in the tannin-rich fraction. The polyphenolic composition of the lingonberry extract was assessed by liquid chromatography-mass spectrometry and was similar to previous reports. The tannin-rich fraction was almost entirely composed of procyanidins of linkage type A and B. Therefore, the antiproliferative activity of lingonberry was caused predominantly by procyanidins. PMID- 18412362 TI - Quali-quantitative analyses of Flavonoids of Morus nigra L. and Morus alba L. (Moraceae) fruits. AB - Morus nigra L., belonging to the Moraceae family, is a decidious tree widely cultivated in Europe and West Asia. It has a long history of medicinal use in Chinese medicine, as a remedy for many kinds of diseases. The aim of the present study was to evaluate the profile of the phenolic constituents of M. nigra fruits and to compare their content with the fruits of another species of Morus, Morus alba, which is also very well known in folklore medicine. The fruits of black and white mulberries have been studied, and five compounds from the methanol extract have been identified by means of HPLC/PDA/ESI-MS. Four compounds (quercetin 3- O glucoside, quercetin 3- O-rutinoside, kaempferol 3- O-rutinoside, and 5- O caffeoylquinic acid) have been isolated by use of Sephadex LH-20 column chromatography and HPLC and characterized by means of NMR and ESI-MS. Furthermore, HPLC/PDA/ESI-MS analysis of the red pigment of M. nigra fruits revealed the presence of four anthocyanins recognized as cyanidin 3- O-glucoside, cyanidin 3- O-rutinoside, pelargonidin 3- O-glucoside, and pelargonidin 3- O rutinoside. All of the compounds were quantified. PMID- 18412363 TI - Comparison of metal oxide-based electronic nose and mass spectrometry-based electronic nose for the prediction of red wine spoilage. AB - Taints caused by Brettanomyces sp. spoilage are of concern to winemakers and consumers. Typically the taints are described as "barnyard", "sweaty saddle", and "Band-aid" when present in red wine at concentrations of several hundred micrograms per liter or more. The two main components of the taint are 4 ethylphenol (4EP) and 4-ethylguaiacol (4EG), which are metabolites produced by Brettanomyces yeasts. There is a need for a rapid instrumental method to quantify these compounds in wines. In this paper are compared two techniques, the metal oxide sensor-based electronic nose (MOS-Enose) and the mass spectrometry-based electronic nose (MS-Enose). Gas chromatography-mass spectrometry (GC-MS) was used for quantification and prediction purposes. Following ethanol removal, the limits of detection of a MOS-Enose were determined as 44 microg L(-1) for 4EP and 91 microg L(-1) for 4EG, using the SY/gCT sensor. These values are significantly lower than the reported human sensory thresholds. Partial least-squares (PLS) regression of electronic nose signals against known levels of 4EP and 4EG in 46 Australian red wines showed that the MOS-Enose was unable to identify "brett" spoilage reliably because of the response of the gas sensors to intersample variation in volatile compounds other than ethylphenols. Conversely, the MS-Enose was capable of reliably estimating concentrations of 4EP higher than 20 microg L( 1). Correlations (r2) of 0.97 and 0.98 were obtained between estimates of 4EP and 4EG concentrations with the concentrations determined by conventional GC-MS. It is concluded that, following ethanol removal, existing metal oxide sensors are sufficiently sensitive to detect brett taints in wine but lack the selectivity needed to perform this task when the aroma volatile background varies. PMID- 18412364 TI - Exposure assessment of prepubertal children to steroid endocrine disruptors. 2. Determination of steroid hormones in milk, egg, and meat samples. AB - In the present study, the occurrence of the main sex steroid hormones in milk, egg, and meat was evaluated on the basis of a highly specific gas chromatography tandem mass spectrometry measurement method. Globally, the results indicated that targeted estrogens and androgens occurred at similar levels (concentration levels in the 10-100 ng kg (-1) range) in the analyzed muscle and milk samples. The same compounds occurred at about 10-fold higher concentrations (i.e., in the 100-1000 ng kg (-1) range) in eggs and kidney samples. More precisely, egg and milk appeared as a non-negligible sources of estradiol (i.e., 2.2 +/- 0.8 and 3.1 +/- 2.0 ng day (-1), respectively), whereas testosterone exposure is caused by ingestion of meat and/or egg (i.e., 12.2 +/- 48.2 and 5.2 +/- 2.3 ng day (-1), respectively). The provided exposure data will be further exploited in the scope of a risk assessment study regarding endocrine disruption associated with these molecules. PMID- 18412365 TI - Influence of thiol metabolism of lactobacilli on egg white proteins in wheat sourdoughs. AB - In wheat sourdoughs, the degradation of gluten proteins is favored by acidification and reducing conditions. This study aimed to determine the proteolytic degradation of egg white proteins in wheat sourdoughs acidified with lactobacilli differing in their thiol metabolism. Ovotransferrin was the only major egg white protein that degraded during sourdough fermentations. An extensive degradation of ovotransferrin required a heterofermentative lactobacilli starter, Lactobacillus sanfranciscensis, with glutathione reductase activity. Ovotransferrin was more resistant to breakdown when sourdoughs were acidified with homofermentative lactobacilli or a mutant strain of L. sanfranciscensis lacking the glutathione reductase. Its susceptibility to proteolysis in L. sanfranciscensis sourdoughs is thus attributable to thiol accumulation by L. sanfranciscensis, which apparently altered the structure of ovotransferrin through a reduction of disulfide bonds. Proteolytic degradation of ovotransferrin was attributable to wheat aspartic proteinases. In addition to the susceptibility to proteolysis, other functional properties of egg proteins may be influenced by thiol-exchange reactions. PMID- 18412366 TI - Spermidine and flavonoid conjugates from peanut (Arachis hypogaea) flowers. AB - A new spermidine triamide derivative has been isolated from peanut flowers and identified as N (1)-acetyl- N (5), N (10)-di- p-( EE)-coumaroylspermidine on the basis of detailed analysis of NMR, MS, and UV data. Two other spermidine conjugates, N (1), N (5), N (10)-tri- p-( EEE)-coumaroylspermidine and di- p-( EE)-coumaroylspermidine, as well as four flavonoid conjugates (quercetin-3 glucoside, quercetin-3-glucuronide, isorhamnetin-3-glucoside, and isorhamnetin-3 glucuronide) that have been previously reported in organs of other plants, have been found in this study in peanut ( Arachis hypogaea L.), a representative of the Leguminosae family, for the first time. The dynamics of photoisomerization in the spermidine conjugates have been investigated. PMID- 18412367 TI - Flavanol and flavonol contents of cocoa powder products: influence of the manufacturing process. AB - Major brands of cocoa powder products present in the Spanish market were analyzed for monomeric flavanols [(+)-catechin and (-)-epicatechin] and flavonols [quercetin-3-glucuronide, quercetin-3-glucoside (isoquercitrin), quercetin-3 arabinoside, and quercetin]. In addition, the influence of the manufacturing process of cocoa powder products, in particular, the alkalinization treatment ( Dutching), on the original content of these flavonoids has been studied. (-) Epicatechin was in the range of 116.02-730.26 microg/g, whereas (+)-catechin was in the range of 81.40-447.62 microg/g in the commercial cocoa products studied. Among flavonols, quercetin-3-arabinoside and isoquercitrin were the major flavonols in the cocoa powder products studied, ranging from 2.10 to 40.33 microg/g and from 3.97 to 42.74 microg/g, respectively, followed by quercetin-3 glucuronide (0.13-9.88 microg/g) and quercetin aglycone (0.28-3.25 microg/g). To our knowledge, these results are the first quantitative data in relation to the content of individualized flavonol derivatives in commercial cocoa powder products. The alkalinization treatment resulted in 60% loss of the mean total flavonoid content. Among flavanols, (-)-epicatechin presented a larger decline (67%, as a mean percentage difference) than (+)-catechin (38%), probably because of its epimerization into (-)-catechin, a less bioavailable form of catechin. A decline was also confirmed for di-, tri-, and tetrameric procyanidins. In the case of flavonols, quercetin presented the highest loss (86%), whereas quercetin 3-glucuronide, quercetin-3-arabinoside, and isoquercitrin showed a similar decrease (58, 62, and 61%, respectively). It is concluded that the large decrease found in the flavonoid content of natural cocoa powder, together with the observed change in the monomeric flavanol profile that results from the alkalinization treatment, could affect the antioxidant properties and the polyphenol biovailability of cocoa powder products. PMID- 18412368 TI - Predicting the clinical lethality of osteogenesis imperfecta from collagen glycine mutations. AB - Osteogenesis imperfecta (OI), or brittle bone disease, often results from missense mutation of one of the conserved glycine residues present in the repeating Gly-X-Y sequence characterizing the triple-helical region of type I collagen. A composite model was developed for predicting the clinical lethality resulting from glycine mutations in the alpha1 chain of type I collagen. The lethality of mutations in which bulky amino acids are substituted for glycine is predicted by their position relative to the N-terminal end of the triple helix. The effect of a Gly --> Ser mutation is modeled by the relative thermostability of the Gly-X-Y triplet on the carboxy side of the triplet containing the substitution. This model also predicts the lethality of Gly --> Ser and Gly --> Cys mutations in the alpha2 chain of type I collagen. The model was validated with an independent test set of six novel Gly --> Ser mutations. The hypothesis derived from the model of an asymmetric interaction between a Gly --> Ser mutation and its neighboring residues was tested experimentally using collagen like peptides. Consistent with the prediction, a significant decrease in stability, calorimetric enthalpy, and folding time was observed for a peptide with a low-stability triplet C-terminal to the mutation compared to a similar peptide with the low-stability triplet on the N-terminal side. The computational and experimental results together relate the position-specific effects of Gly --> Ser mutations to the local structural stability of collagen and lend insight into the etiology of OI. PMID- 18412369 TI - Residence time of receptor-ligand complexes and its effect on biological function. AB - The formation and duration of binary receptor-ligand complexes are fundamental to many physiologic processes. Most often, the effectiveness of interaction between a receptor and its ligand is quantified in terms of closed system, equilibrium affinity measurements, such as IC50 and Kd. In the context of in vivo biology, however, the extent and duration of responses to receptor-ligand interactions depend greatly on the time period over which the ligand is in residence on its receptor. Here we define receptor-ligand complex residence time in quantitative terms and describe its significance to biological function. Examples of the importance of residence time are presented for natural ligands of different receptor types. The impact of residence time on the optimization of potential ligands as drugs for human medicine is also described. PMID- 18412370 TI - Two-dimensional X-ray photoelectron spectroscopy for composite surface analysis. AB - We describe a method for obtaining two-dimensional X-ray photoelectron spectroscopic data derived from the frequency dependence of the XPS peaks recorded under electrical square-wave pulses, which control and affect the binding energy positions via the electrical potentials developed as a result of charging. By using cross-correlations between various peaks, our technique enables us to elucidate electrical characteristics of surface structures of composite samples and bring out various correlations between hidden/overlapping peaks. PMID- 18412371 TI - Identification of surface heterogeneity effects in cyclic voltammograms derived from analysis of an individually addressable gold array electrode. AB - Voltammetric behavior at gold electrodes in aqueous media is known to be strongly dependent on electrode polishing and history. In this study, an electrode array consisting of 100 nominally identical and individually addressable gold disks electrodes, each with a radius of 127 microm, has been fabricated. The ability to analyze both individual electrode and total array performance enables microscopic aspects of the overall voltammetric response arising from variable levels of inhomogeneity in each electrode to be identified. The array configuration was initially employed with the reversible and hence relatively surface insensitive [Ru(NH 3) 6] (3+/2+) reaction and then with the more highly surface sensitive quasi-reversible [Fe(CN) 6] (3-/4-) process. In both these cases, the reactants and products are solution soluble and, at a scan rate of 50 mV s (-1), each electrode in the array is assumed to behave independently, since no evidence of overlapping of the diffusion layers was detected. As would be expected, the variability of the individual electrodes' responses was significantly larger than found for the summed electrode behavior. In the case of cytochrome c voltammetry at a 4,4'-dipyridyl disulfide modified electrode, a far greater dependence on electrode history and electrode heterogeneity was detected. In this case, voltammograms derived from individual electrodes in the gold array electrode exhibit shape variations ranging from peak to sigmoidal. However, again the total response was always found to be well-defined. This voltammetry is consistent with a microscopic model of heterogeneity where some parts of each chemically modified electrode surface are electroactive while other parts are less active. The findings are consistent with the common existence of electrode heterogeneity in cyclic voltammetric responses at gold electrodes, that are normally difficult to detect, but fundamentally important, as electrode nonuniformity can give rise to subtle forms of kinetic and other forms of dispersion. PMID- 18412372 TI - Charge-coupled device operated in a time-delayed integration mode as an approach to high-throughput flow-based single molecule analysis. AB - Single molecule detection (SMD) readouts are particularly attractive for assays geared toward high-throughput processing, because they can potentially reduce assay time by eliminating various processing steps. Unfortunately, most flow based SMD experiments have generated low throughputs due primarily to the fact that they are configured in single assay formats. The use of a charge-coupled device (CCD) with flow-based SMD can image multiple single molecule assays simultaneously to realize high-throughput processing capabilities. We present, for the first time, the ability to simultaneously track and detect single molecules in multiple microfluidic channels by employing a CCD camera operated in time-delayed integration (TDI) mode as a means for increasing the throughput of any single molecule measurement. As an example of the technology, we have configured a CCD to operate in a TDI mode to detect single double-stranded DNA molecules (lambda and pBR322) labeled with an intercalating dye (TOTO-3) in a series of microfluidic channels poised on a poly(methyl methacrylate), PMMA, chip. A laser beam was launched into the side of the chip, which irradiated a series of fluidic channels (eight) with the resulting fluorescence imaged onto a CCD. Using this system, we were able to identify single DNA molecules based on the fluorescence burst intensity arising from differences in the extent of dye labeling associated with the DNA molecule length. The CCD/TDI approach allowed increasing sample throughput by a factor of 8 compared to a single-assay SMD experiment. A sampling throughput of 276 molecules s (-1) per channel and 2208 molecules s (-1) for an eight channel microfluidic system was demonstrated. Operated in its full capacity, this multichannel format was projected to yield a sample throughput of 1.7 x 10 (7) molecules s (-1), which represents a 170-fold improvement over previously reported single molecule sampling rates. PMID- 18412373 TI - Beta-hairpin peptidomimetics: design, structures and biological activities. AB - The folded 3D structures of peptides and proteins provide excellent starting points for the design of synthetic molecules that mimic key epitopes (or surface patches) involved in protein-protein and protein-nucleic acid interactions. Protein epitope mimetics (PEMs) may recapitulate not only the structural and conformational properties of the target epitope but also their biological activities. By transferring the epitope from a recombinant to a synthetic scaffold that can be produced by parallel combinatorial methods, it is possible to optimize properties through iterative cycles of library synthesis and screening, and even to evolve new biological activities. One very interesting scaffold is found in beta-hairpin motifs, which are used by many proteins to mediate molecular recognition events. This motif is readily amenable to PEM design, for example, by transplanting hairpin loop sequences from folded proteins onto hairpin-stabilizing templates, such as the dipeptide D-Pro-L-Pro. In addition, beta-hairpin peptidomimetics can also be exploited to mimic other types of epitopes, such as those based on alpha-helical secondary structures. The size and shape of beta-hairpin PEMs appear well suited for the design of inhibitors of both protein-protein and protein-nucleic acid interactions, endeavors that have so far proven difficult using small "drug-like" molecules. In recent work, it was shown that beta-hairpin PEMs can be designed that mimic the canonical conformations of antibody hypervariable loops, suggesting that novel small molecule antibody mimics may be feasible. Using naturally occurring peptides as starting points, beta-hairpin mimetics have been discovered that possess antimicrobial activity, while others are potent inhibitors of the chemokine receptor CXCR4. Beta-hairpin PEMs have also been designed and optimized that mimic an alpha-helical epitope in p53 and so block its interaction with HDM2. A crystal structure of one HDM2-mimetic complex revealed how the surface of the protein had adapted to the shape of the hairpin, thereby enhancing inhibitor affinity. Small folded RNA motifs also make interesting targets for inhibitor design. For example, beta-hairpin mimetics have been designed and optimized that bind with high affinity and good selectivity to the TAR and RRE RNA motifs from HIV-1. Solution structures of the mimetics both free and bound to the RNA target provided some surprises, as well as an improved understanding of the mechanisms of binding. These mimetics represent still a relatively new family of RNA-binding molecules, but clearly one with potential for development into novel antiviral agents. PMID- 18412374 TI - Synthesis of ellipsoidal hematite/silica/polymer hybrid materials and the corresponding hollow polymer ellipsoids. AB - Ellipsoidal trilayer hematite/silica/poly(divinylbenzene) hybrid particles were prepared by distillation precipitation polymerization of divinyl benzene (DVB) in the presence of hematite/3-(methacryloxy)propyl trimethoxysilane (MPS)-modified silica (SiO(2)) core-shell particles as the seeds. The polymerization of DVB was performed in neat acetonitrile with 2,2'-azobisisobtyronitrile (AIBN) as initiator to coat the hematite/MPS-modified SiO(2) seeds through the capture of DVB oligomer radicals with the aid of a vinyl group on the surface of the hematite/MPS-modified silica core-shell particles in the absence of any stabilizer or surfactant. The other hematite/silica/polymer trilayer hybrid particles with different polarity and various functionality, such as hematite/silica/poly(ethylene glycol dimethacrylate) and hematite/silica/poly(divinyl benzene- co-methacrylic acid) could also be prepared by this procedure. Hematite/silica/poly( N, N'-methylenebisacrylamide) composite particles could be prepared with unmodified hematite/silica particles as seeds. Hollow poly(divinyl benzene) (PDVB) and poly( N, N'-methylenebisacrylamide) (PMBAAm) ellipsoids with movable hematite cores were subsequently developed after the selective etching of the silica midlayer in diluted hydrofluoric acid from hematite/silica/PDVB and hematite/silica/PMBAAm trilayer hybrids. Hollow PDVB ellipsoids were obtained by removal of the silica midlayer and hematite core of the trilayer hybrids with concentrated HF solution. The resultant trilayer hybrid particles and hollow polymer ellipsoids were characterized by transmission electron microscopy and vibrating sample magnetometer. PMID- 18412375 TI - Near-infrared emission from novel Tris(8-hydroxyquinolinate)lanthanide(III) complexes-functionalized mesoporous SBA-15. AB - A novel mesoporous material covalently bonded with 8-hydroxyquinoline (HQ) was synthesized (designated as Q-SBA-15). The 5-formyl-8-hydroxyquinoline grafted to (3-aminopropyl)triethoxysilane, that is, alkoxysilane modified 8-hydroxyquinoline (Q-Si), was used as one of the precursors for the preparation of the Q-SBA-15 material. On the basis of the other function of the Q-Si of coordinating to lanthanide (Ln) ions, for the first time, the LnQ 3 complexes (Ln = Er, Nd, Yb) have been covalently bonded to the SBA-15 materials. The derivative materials, denoted as LnQ 3-SBA-15, were characterized by field emission scanning electron microscopy (FE-SEM), powder X-ray diffraction (XRD), transmission electron microscopy (TEM), nitrogen adsorption-desorption, and fluorescence spectra. Upon excitation at the ligands absorption bands, all of these materials show the characteristic near-infrared (NIR) luminescence of the corresponding lanthanide ions through the intramolecular energy transfer from the ligands to the lanthanide ions. The NIR luminescence of these mesoporous materials was compared with that of the corresponding pure LnQ 3 complexes and discussed in detail. PMID- 18412376 TI - Reduced hydrophobic interaction of polystyrene surfaces by spontaneous segregation of block copolymers with oligo (ethylene glycol) methyl ether methacrylate blocks: force measurements in water using atomic force microscope with hydrophobic probes. AB - Reduction of hydrophobic interaction in water is important in biological interfaces. In our previous work, we have found that poly(styrene- b-triethylene glycol methyl ether methacrylate) (PS-PME3MA) segregates the PME3MA block to the surface in hydrophobic environment, such as in air or in a vacuum, and shows remarkable resistance against adsorption or adhesion of proteins, platelets, and cells in water. In this paper, we report that atomic force microscopy (AFM) with hydrophobic probes can directly monitor the reduced hydrophobic interaction of the PS surfaces modified by poly(styrene- b-origoethylene glycol methyl ether methacrylate) (PS-PME NMA), where N is the number of ethylene glycol units. The pull-off forces between the hydrophobic probes that are coated with octyltrichlorosilane (OLTS) and the PS-PME NMA modified polystyrene (PS) surfaces in water were measured. The absolute spring constants and tip-curvatures of the AFM cantilevers were measured to compute the work of adhesion by the Johnson, Kendall, and Roberts (JKR) theory, which relates the pull-off force at which the separation occurs between a hemisphere and a plane to the work of adhesion. The hydrophobic interactions between the hydrophobic tip and polymer surfaces in water were greatly reduced with the segregated PME NMA blocks. The hydrophobic interactions decrease with increasing N of the series of PS-PME NMA and show a correlation with the amount of protein adsorbed. PMID- 18412377 TI - On the mechanism of low-pressure imprint lithography: capillarity vs viscous flow. AB - Dominant mechanisms in low-pressure imprint lithography processes have been identified for the regimes that are definable in terms of applied pressure, temperature, and mold material characteristics. Capillarity is found to be the dominant mechanism at high temperature and low pressure when stiff, hard molds are used. In the case of flexible thin-film ( approximately 20 microm) molds, both the capillarity and the viscous flow are involved. Both mechanisms are operative in the initial stage of the imprinting, but the capillarity takes over as time progresses. PMID- 18412378 TI - Multidentate surface ligand exchange for the immobilization of CdSe/ZnS quantum dots and surface quantum dot-oligonucleotide conjugates. AB - A method for synthesizing multidentate thiol ligands on fused silica surfaces (e.g., optical fibers) was developed for the immobilization of CdSe/ZnS quantum dots (QDs) capped with hydrophilic or hydrophobic ligands. This work was motivated by the poor stability of QDs immobilized via monodentate thiol ligands and the need for stable immobilization strategies in the development of sensor technologies based on QDs. Multi-dentate immobilization was able to withstand washing protocols, and surface ligand exchange occurred via self-assembly through the zinc-metal affinity interaction. Atomic force and scanning electron microscopy images suggested that the QDs were immobilized at high density, approximately 2-4 x 10 (13) cm (-2). It was possible to immobilize one, two, or three colors of QD. Upon immobilization, 1-2 nm bathochromic shifts in the PL spectra were observed. This was attributed to both ligand exchange and the change in local environment. The change in environment was accompanied by a decrease in PL lifetime. Self-assembly of immobilized QD-oligonucleotide and QD-avidin conjugates was also demonstrated. These conjugates were able to hybridize with complementary oligonucleotide and bind biotin, respectively. This versatile immobilization chemistry is an important step in the development of surface-based QD nanosensors. Such technology requires QDs to be immobilized such that they remain accessible to target molecules in solution. PMID- 18412379 TI - Synthesis and optical characterization of submicrometer gold nanotubes grown on goethite rods. AB - Goethite (FeOOH) rods were used as templates for growing gold nanotubes with a length of a few hundred nanometers and an aspect ratio between 3 and 4. Successful uniform growth required surface modification, followed by the attachment of small Au seeds and one-step seeded growth using formaldehyde as a reducing agent, as previously reported for the growth of Au shells on silica spheres and hematite spindles. The thickness and surface roughness of the obtained shells could be adjusted by simply varying the concentration ratio between seeds (modified goethite rods) and growth reagents (HAuCl 4 and formaldehyde). The morphology of the synthesized gold nanotubes was thoroughly characterized by TEM, SEM, and AFM/MFM. The resulting gold nanotubes display well defined plasmon resonances, with a strong longitudinal mode centered around ca. 1400 nm and a broad band in the visible resulting from the overlap of a transverse mode and a multipolar mode, as was found from theoretical modeling using the boundary element method, which provides reasonable agreement with the experimental results. PMID- 18412380 TI - Evidence for effect of GM1 on opioid peptide conformation: NMR study on leucine enkephalin in ganglioside-containing isotropic phospholipid bicelles. AB - Enkephalins are endogenous neuropeptides that have opioid-like activities and compete with morphines for the receptor binding. The binding of these neuropeptides to membrane appears crucial since enkephalins interact with the nerve cell membranes to achieve bioactive conformations that fit onto multiple receptor sites (micro, delta, and kappa). Using NMR spectroscopy, we have determined the solution structure of the small opiate pentapeptide leucine enkephalin in the presence of isotropic phospholipid bicelles: phosphocholine bicelles (DMPC:CHAPS 1:4) and phosphocholine bicelles doped with ganglioside GM1 (DMPC:CHAPS:GM1 1:4:0.3). Bicelles containing GM1 were found to interact strongly with leucine enkephalin, whereas a somewhat weaker interaction was observed in the case of bicelles without GM1. Structure calculation from torsion angles, chemical shifts, and NOE-based distance constraints explored that the peptide could flexibly switch between several mu- and delta-selective conformations in both the bicelles though micro-selective conformations turned out to be geometrically preferred in each bicellar system. A detailed analysis of the structures presented supports the variance over the singly associated conformation of enkephalin in nerve cell membranes. PMID- 18412381 TI - Water in nanoconfinement between hydrophilic self-assembled monolayers. AB - Molecular dynamics (MD) simulations of water confined to subnanometer thicknesses between carboxyl-terminated alkanethiol self-assembled monolayers (SAMs) on gold were performed to address conflicts in the literature on the structure and response of water in confinement. The amount of water was varied to yield submonolayer to bilayer structures. The orientation of the water is affected by the confinement, especially in the submonolayer case. We find that the diffusion coefficient decreases as the film becomes thinner and at higher pressures. However, in all cases studied, liquid diffusion is always found. At maximal suppression, the diffusion constant is 2 orders of magnitude smaller than the bulk value. PMID- 18412382 TI - Modified ligand-exchange for efficient solubilization of CdSe/ZnS quantum dots in water: a procedure guided by computational studies. AB - One of the methods to render CdSe/ZnS core-shell quantum dots(QDots) water soluble is to functionalize the surface with carboxylate groups by the use of heterobifunctional ligands such as 3-mercaptopropionic acid, where the thiolic end binds onto the outer ZnS shell. However, currently available ligand-exchange procedures starting with TOPO-capped quantum dots often lead to significant loss of quantum yields and poor stability of the colloids in water. As part of our efforts to overcome these problems, we used computational methods to understand the nature of binding between alkyl thiols and ZnS wurtzite surfaces. Guided by the computational results, we modified the ligand-exchange method and increased the reactivity of 3-mercaptopropionic acid toward the ZnS surface in chloroform. The functionlization reaction required only mild reaction conditions and led to QDot nanoparticles that were individually dispersed in water with good colloidal stability. Importantly, the photoluminescence performance of the QDots was highly preserved. PMID- 18412383 TI - The influence of sample preparation and replicate analyses on HeLa Cell phosphoproteome coverage. AB - Ongoing optimization of proteomic methodologies seeks to improve both the coverage and confidence of protein identifications. The optimization of sample preparation, inclusion of technical replicates (repeated instrumental analysis of the same sample), and biological replicates (multiple individual samples) are crucial in proteomic studies to avoid the pitfalls associated with single point analysis and under-sampling. Phosphopeptides were isolated from HeLa cells and analyzed by nano-reversed phase liquid chromatography electrospray ionization tandem mass spectrometry (nano-RP-LC-MS/MS). We observed that a detergent-based protein extraction approach, followed with additional steps for nucleic acid removal, provided a simple alternative to the broadly used Trizol extraction. The evaluation of four technical replicates demonstrated measurement reproducibility with low percent variance in peptide responses at approximately 3%, where additional peptide identifications were made with each added technical replicate. The inclusion of six technical replicates for moderately complex protein extracts (approximately 4000 uniquely identified peptides per data set) affords the optimal collection of peptide information. PMID- 18412384 TI - Different signaling pathways expressed by chicken naive CD4(+) T cells, CD4(+) lymphocytes activated with staphylococcal enterotoxin B, and those malignantly transformed by Marek's disease virus. AB - Proteomics methods, based on liquid chromatography and tandem mass spectrometry, produce large "shotgun" proteomes that are most appropriately compared not at the level of differentially expressed proteins only but at the more comprehensive level of biological networks and pathways. This is now possible with the emergence of functional annotation databases and tools, databases of canonical pathways and molecular interactions and computational text mining tools. Here, we used shotgun proteomics, and the differential proteomics modeling functionalities available in the Pathwaystudio network modeling program to define the cell physiology of Hodgkin's disease antigen-overexpressing (CD30 (hi)) CD4 (+) T cell lymphomas using the unique Marek's disease (MD) natural animal model. CD30 (hi) lymphoma cells have characteristics of activated T cells but are also fundamentally different from their nontransformed healthy counterparts. We compared the cell physiology of naive, superantigen-activated and MD-transformed CD4 (+) T cell proteomes. While the superantigen-activated cells had signaling pathways associated with cell activation, inflammation, proliferation and cell death, the MD-transformed cells had growth factor, cytokine, adhesion, and transcription factor signaling responses associated with oncogenicity, cell proliferation, angiogenesis, motility, and metastasis. PMID- 18412385 TI - Label-free detection of differential protein expression by LC/MALDI mass spectrometry. AB - Protein abundance changes during disease or experimental perturbation are increasingly analyzed by label-free LC/MS approaches. Here we demonstrate the use of LC/MALDI MS for label-free detection of protein expression differences using Escherichia coli cultures grown on arabinose, fructose or glucose as a carbon source. The advantages of MALDI, such as detection of only singly charged ions, and MALDI plate archiving to facilitate retrospective MS/MS data collection are illustrated. MALDI spectra from RP chromatography of tryptic digests of the E. coli lysates were aligned and quantitated using the Rosetta Elucidator system. Approximately 5000 peptide signals were detected in all LC/MALDI runs spanning over 3 orders of magnitude of signal intensity. The average coefficients of variation for all signals across the entire intensity range in all technical replicates were found to be <25%. Pearson correlation coefficients from 0.93 to 0.98 for pairwise comparisons illustrate high replicate reproducibility. Expression differences determined by Analysis of Variance highlighted over 500 isotope clusters ( p < 0.01), which represented candidates for targeted peptide identification using MS/MS. Biologically interpretable protein identifications that could be derived underpin the general utility of this label-free LC/MALDI strategy. PMID- 18412386 TI - Room-temperature Kumada cross-coupling of unactivated aryl chlorides catalyzed by N-heterocylic carbene-based nickel(II) complexes. AB - The Kumada cross-coupling reaction of a variety of unactivated aryl chlorides, vinyl chlorides, and heteroaryl chlorides catalyzed by nickel(II) complexes containing pyridine-functionalized NHC ligands is described. The catalysts are so active that the reactions proceed at room temperature in excellent yields. PMID- 18412387 TI - Phorbasides A-E, cytotoxic chlorocyclopropane macrolide glycosides from the marine sponge Phorbas sp. CD determination of C-methyl sugar configurations. AB - Five new cytotoxic macrolide glycosides phorbasides A-E (3-7), each possessing a macrolide ring appended to a rare ene-yne-trans-2-chlorocyclopropane, were isolated from the same Western Australian sponge (Phorbas sp.) that provided phorboxazoles A and B. The structures of 3-7 were solved by analysis of spectroscopic data including NMR, MS, and CD. A synthesis of methyl 2-O-methyl alpha-L-evalose from L-rhamnose was completed and used for configurational assignment of the sugar residue in 3. Acid-catalyzed methanolysis of 3 followed by two-step derivatization of the liberated O-methyl glycoside gave a vicinal 4-O naphthoyl/tertiary 3-N-(2-aminonaphthyl)carbamate derivative that exhibited exciton coupled CD identical with that of the derivative prepared from synthetic 1,2- O-dimethyl-alpha-L-evalose. PMID- 18412388 TI - Efficient synthesis of cis- and trans-3,4-dihydroxy-3,4-dihydromollugin. AB - An efficient synthesis of naturally occurring compounds isolated from Pentas longiflora, cis-3,4-dihydroxy-3,4-dihydromollugin 2, and trans-3,4-dihydroxy-3,4 dihydromollugin 3 is described. The O-protected mollugins were dihydroxylated using OsO4 to achieve the corresponding cis-dihydroxy derivatives in excellent yield. The synthesis of trans-3,4-dihydroxy-3,4-dihydromollugin was achieved using Oxone in good yield. A mechanism for the formation of cis-3,4 dihydroxymollugin acetonide from the reaction of mollugin with Oxone is proposed. PMID- 18412389 TI - Functionalization of organotrifluoroborates: reductive amination. AB - Herein we report the conversion of aldehyde-containing potassium and tetrabutylammonium organotrifluoroborates to the corresponding amines through reductive amination protocols. Potassium formate facilitated by catalytic palladium acetate, sodium triacetoxyborohydride, and pyridine borane have all served as effective hydride donors, reducing the initially formed imines or iminium ions to provide the corresponding amines. PMID- 18412390 TI - Efficient synthesis of trifluoromethyl and related trisubstituted alkene dipeptide isosteres by palladium-catalyzed carbonylation of amino acid derived allylic carbonates. AB - A novel stereoselective synthetic approach to (Z)-trifluoromethylalkene dipeptide isosteres (CF(3)-ADIs) is described. Starting from readily available N-Boc-L phenylalanine, Phe-Gly type CF(3)-ADIs were obtained through palladium-catalyzed carbonylation of allylic carbonates under CO. While the reaction of N-Boc derivatives proceeds in excellent yields but lower stereoselectivity (E: Z = 62:38-43:57), the reaction of the N, N-diBoc derivative exclusively affords the desired (Z)-isomer in 61% yield. We also present a highly stereoselective synthesis of several Phe-Gly type trisubstituted alkene dipeptide isosteres by palladium-catalyzed carbonylation. PMID- 18412391 TI - Physical organic chemistry on the brain. AB - The challenges to obtaining chemical-scale information on the molecules of neuroscience are considerable. Most targets are complex integral membrane proteins that are not amenable to direct structural characterization. However, by combining the tools of organic synthesis, molecular biology, and electrophysiology, rational and systematic structure-function studies can be performed in what we have termed physical organic chemistry on the brain. Using these tools, we have probed hydrophobic effects, hydrogen bonding, cation-pi interactions, and conformational changes associated with channel gating. The insights gained provide important guidance for drug discovery efforts targeting ion channels and neuroreceptors and mechanistic insights for the complex proteins of neuroscience. PMID- 18412392 TI - A convenient route to N-[2-(Fmoc)aminoethyl]glycine esters and PNA oligomerization using a Bis-N-Boc nucleobase protecting group strategy. AB - A simple and practical synthesis of the benzyl, allyl, and 4-nitrobenzyl esters of N-[2-(Fmoc)aminoethyl]glycine is described starting from the known N-(2 aminoethyl)glycine. These esters are stored as stable hydrochloride salts and were used in the synthesis of peptide nucleic acid monomers possessing bis-N-Boc protected nucleobase moieties on the exocyclic amino groups of ethyl cytosin-1 ylacetate, ethyl adenin-9-ylacetate and ethyl (O(6)-benzylguanin-9-yl)acetate. Upon ester hydrolysis, the corresponding nucleobase acetic acids were coupled to N-[2-(Fmoc)aminoethyl]glycine benzyl ester or to N-[2-(Fmoc)aminoethyl]glycine allyl ester in order to retain the O(6) benzyl ether protecting group of guanine. The Fmoc/bis-N-Boc-protected monomers were successfully used in the Fmoc-mediated solid-phase peptide synthesis of mixed sequence 10-mer PNA oligomers and are shown to be a viable alternative to the currently most widely used Fmoc/Bhoc protected peptide nucleic acid monomers. PMID- 18412393 TI - Phenolic glycosides from the stem bark of Caryocar villosum and C. glabrum. AB - Mushroom tyrosinase inhibitory activity of methanol extracts and polar fractions of the stem bark of Caryocar villosum and C. glabrum has been assessed. Seven new phenolic glycosides (1-7) were isolated from the most active fractions, along with 15 known compounds (8-22). The structures of these compounds were established on the basis of spectroscopic methods including 1D and 2D NMR analysis, HRESIMS, and comparison with literature experimental data for known compounds. PMID- 18412394 TI - Effect of the major glycosides of Harpagophytum procumbens (Devil's Claw) on epidermal cyclooxygenase-2 (COX-2) in vitro. AB - Harpagophytum procumbens, commonly known as Devil's Claw, is indigenous to southern Africa, and extracts of the tubers have been used for centuries in the treatment of a variety of inflammatory disorders. Its major active components, harpagoside (1), harpagide (2), 8-coumaroylharpagide (3), and verbascoside (4), are believed to interact either synergistically or antagonistically in modulating the enzymes responsible for inducing inflammation, although this has not been probed hitherto. In the current work, the ability of these compounds to inhibit the expression of COX-2 following administration to freshly excised porcine skin has been investigated. An ethanol-soluble extract of H. procumbens tubers and two of the pure compounds tested showed promising activity in Western blotting and immunocytochemical assays, with harpagoside (1) and 8-coumaroylharpagide (3) exhibiting greater reductions in COX-2 expression than verbascoside (4). Harpagide (2) caused a significant increase in the levels of COX-2 expression after 6 h of topical application. The data suggest that the efficacy of H. procumbens is dependent upon the ratios of compounds 1-4 present, which is inconsistent with some current official monograph specifications based solely on harpagoside (1) content. PMID- 18412395 TI - Corymbones A and B, phloroglucinols with thyrotropin releasing hormone receptor 2 binding affinity from the flowers of Corymbia peltata. AB - High-throughput screeing of a plant and marine invertebrate extract library to find natural products with rat thytotropin releasing hormone receptor 2 binding affinity led to the isolation of two new active acylphloroglucinols, corymbones A and B (1 and 2) from flowers of the Queensland tree Corymbia peltata. Their structures were assigned from interpretation of 2D NMR and high-resolution ESIMS data. Compounds 1 and 2 showed rat TRH receptor 2 binding affinity with IC 50 values of 23 and 19 microM, respectively. PMID- 18412396 TI - Itosides J-N from Itoa orientalis and structure - anti-COX-2 activity relationship of phenolic glycosides. AB - Two new phenolic glycosides, itosides J (1) and K (2), two new cylcohexenoyl glycosides, itosides L (3) and M (4), a new flavone glycoside, itoside N (5), and echitin (6) were isolated from the extract of the bark, twigs, and leaves of Itoa orientalis, together with 22 known compounds. The structures were elucidated by means of UV, IR, MS, and NMR techniques, and the relative configuration of compound 3 was confirmed by X-ray crystallography. NMR data for 6 are reported for the first time. Compounds 1, 3, 5, and phenolic glycosides 7- 22 were also assayed for anti-inflammatory activity against COX-2. Compounds 8, 10, 12- 14, 16, 19, 24, 26, and 27 showed significant inhibitory effects, with inhibitory rates of 49.7-85.3% at 10 microM. PMID- 18412397 TI - Alkaloids from the Australian rainforest tree Ochrosia moorei. AB - High-throughput screening of a plant and marine invertebrate extract library to find natural products that down-regulate expression of pro-inflammatory genes associated with the glucocorticoid receptor ligand complex led to the identification of bioactive CH2Cl 2 extracts from stems and leaves of the Queensland tree Ochrosia moorei. Bioassay-guided purification of the stem extract enabled the isolation of four alkaloids including two new compounds, ochrosamines A (1) and B (2), and the known compounds ellipticine (3) and 9-methoxyellipticine (4). The leaf extract also afforded 3 and 4 as well as apparicine (5) and desoxycordifoline (6). The structures of the two new compounds were assigned from interpretation of 2D NMR and high-resolution ESIMS data. Ellipticine and 9 methoxyellipticine were the most active components, and both displayed IC 50 values of 90 microM. Apparicine and desoxycordifoline were only very weakly active, and ochrosamines A and B were inactive. PMID- 18412398 TI - Scopularides A and B, cyclodepsipeptides from a marine sponge-derived fungus, Scopulariopsis brevicaulis. AB - Two novel cyclodepsipeptides, scopularides A and B, were found in the fungus Scopulariopsis brevicaulis, which was isolated from the marine sponge Tethya aurantium. In addition, the known fungal metabolite paxilline was identified. The structures of the scopularides were elucidated by NMR, MS, and chemical derivatization methods as cyclo-(3-hydroxy-4-methyldecanoyl-Gly-L-Val-D-Leu-L-Ala L-Phe) and cyclo-(3-hydroxy-4-methyloctanoyl-Gly-L-Val-D-Leu-L-Ala-L-Phe) for scopularide A and B, respectively. Antibiotic activity against Gram-negative bacteria was absent and against Gram-positive bacteria was weak, but activity against several tumor cell lines was significant at 10 microg/mL. PMID- 18412399 TI - Microwave-assisted combinatorial synthesis of hexa-substituted 1,4 dihydropyridines scaffolds using one-pot two-step multicomponent reaction followed by a S-alkylation. AB - A sequential one-pot two-step protocol for microwave-assisted synthesis of Hantzsch-type hexa-substituted 1,4-dihydropyridines has been developed. The three component reactions of beta-aroylthioamides with aldehydes and acetonitrile derivatives produce the intermediates in situ followed by a S-alkylation to afford hexa-substituted 1,4-dihydropyridines. The reaction presumably proceeds via a Knoevenagel condensation-Michael addition-cyclocondensation-rearrangement-S N2 reaction sequence. Target compounds were obtained in high yields and simply purified by recrystallization. The novel method is complementary to the classical Hantzsch synthesis in that it is well-suited to the preparation of hexa substituted 1,4-dihydropyridines. PMID- 18412400 TI - Biomimetic interfaces for high-performance optics in the deep-UV light range. AB - We report an innovative approach for the fabrication of highly light transmissive, antireflective optical interfaces. This is possible due to the discovery that metallic nanoparticles may be used as a lithographic mask to etch nonstraightforward structures into fused silica, which results in a quasihexagonal pattern of hollow, pillar-like protuberances. The far reaching optical performance of these structures is demonstrated by reflection and transmission measurements at oblique angles of incidence over a broad spectral region ranging from deep-ultraviolet to infrared light. PMID- 18412401 TI - A nonvolatile plasmonic switch employing photochromic molecules. AB - We demonstrate a surface plasmon-polariton (SPP) waveguide all-optical switch that combines the unique physical properties of small molecules and metallic (plasmonic) nanostructures. The switch consists of a pair of gratings defined in an aluminum film coated with a 65 nm thick layer of photochromic (PC) molecules. The first grating couples a signal beam consisting of free space photons to SPPs that interact effectively with the PC molecules. These molecules can reversibly be switched between transparent and absorbing states using a free space optical pump. In the transparent (signal "on") state, the SPPs freely propagate through the molecular layer, and in the absorbing (signal "off") state, the SPPs are strongly attenuated. The second grating serves to decouple the SPPs back into a free space optical beam, enabling measurement of the modulated signal with a far field detector. In a preliminary study, the switching behavior of the PC molecules themselves was confirmed and quantified by surface plasmon resonance spectroscopy. The excellent (16%) overlap of the SPP mode profile with the thin layer of switching molecules enabled efficient switching with power densities of approximately 6.0 mW/cm2 in 1.5 microm x 8 microm devices, resulting in plasmonic switching powers of 0.72 nW per device. Calculations further showed that modulation depths in access of 20 dB can easily be attained in optimized designs. The quantitative experimental and theoretical analysis of the nonvolatile switching behavior in this letter guides the design of future nanoscale optically or electrically pumped optical switches. PMID- 18412402 TI - Modulation of in vivo tumor radiation response via gold nanoshell-mediated vascular-focused hyperthermia: characterizing an integrated antihypoxic and localized vascular disrupting targeting strategy. AB - We report noninvasive modulation of in vivo tumor radiation response using gold nanoshells. Mild-temperature hyperthermia generated by near-infrared illumination of gold nanoshell-laden tumors, noninvasively quantified by magnetic resonance temperature imaging, causes an early increase in tumor perfusion that reduces the hypoxic fraction of tumors. A subsequent radiation dose induces vascular disruption with extensive tumor necrosis. Gold nanoshells sequestered in the perivascular space mediate these two tumor vasculature-focused effects to improve radiation response of tumors. This novel integrated antihypoxic and localized vascular disrupting therapy can potentially be combined with other conventional antitumor therapies. PMID- 18412403 TI - Quantum dot-fluorescent protein pairs as novel fluorescence resonance energy transfer probes. AB - Fluorescence resonance energy transfer (FRET) characteristics, including the efficiency, donor-acceptor distance, and binding strength of six fluorescent protein (FP)-quantum dot (QD) pairs were quantified, demonstrating that FPs are efficient acceptors for QD donors with up to 90% quenching of QD fluorescence and that polyhistidine coordination to QD core-shell surface is a straightforward and effective means of conjugating proteins to commercially available QDs. This provides a novel approach to developing QD-based FRET probes for biomedical applications. PMID- 18412404 TI - Defining process design space for biotech products: case study of Pichia pastoris fermentation. AB - The concept of "design space" has been proposed in the ICH Q8 guideline and is gaining momentum in its application in the biotech industry. It has been defined as "the multidimensional combination and interaction of input variables (e.g., material attributes) and process parameters that have been demonstrated to provide assurance of quality." This paper presents a stepwise approach for defining process design space for a biologic product. A case study, involving P. pastoris fermentation, is presented to facilitate this. First, risk analysis via Failure Modes and Effects Analysis (FMEA) is performed to identify parameters for process characterization. Second, small-scale models are created and qualified prior to their use in these experimental studies. Third, studies are designed using Design of Experiments (DOE) in order for the data to be amenable for use in defining the process design space. Fourth, the studies are executed and the results analyzed for decisions on the criticality of the parameters as well as on establishing process design space. For the application under consideration, it is shown that the fermentation unit operation is very robust with a wide design space and no critical operating parameters. The approach presented here is not specific to the illustrated case study. It can be extended to other biotech unit operations and processes that can be scaled down and characterized at small scale. PMID- 18412406 TI - Free radical conformations and conversions in X-irradiated single crystals of L cysteic acid by electron magnetic resonance and density functional theory studies. AB - Single crystals of L-cysteic acid monohydrate were X-irradiated and studied at 295 K using EPR, ENDOR, and EIE techniques. Three spectroscopically different radicals were observed. These were a deamination radical reduction product (R1), and two oxidation products formed by hydrogen abstraction (radicals R2, R3). R2 and R3 were shown to exhibit the same chemical structure while exhibiting very different geometrical conformations. Cluster DFT calculations at the 6-31G(d,p) level of theory supported the experimental observations for radicals R1 and R2. It was not possible to simulate the R3 radical in any attempted cluster; hence, for this purpose a single molecule approach was used. The precursor radicals for R1, R2, and R3, identified in the low-temperature work on L-cysteic acid monohydrate by Box and Budzinski, were also investigated using DFT calculations. The experimentally determined EPR parameters for the low-temperature decarboxylated cation could only be reproduced correctly within the cluster when the carboxyl group remained in the proximity of the radical. Only one of the two observed low-temperature carboxyl anions (stable at 4 and 48 K) could be successfully simulated by the DFT calculations. Evidence is presented in support of the conclusions that the carboxyl reduction product already is protonated at 4 K and that the irreversible conversion between the two reduction products is brought forward by an umbrella-type inversion of the carboxyl group. PMID- 18412405 TI - Role of network branching in eliciting differential short-term signaling responses in the hypersensitive epidermal growth factor receptor mutants implicated in lung cancer. AB - We study the effects of EGFR inhibition in wild-type and mutant cell lines upon tyrosine kinase inhibitor TKI treatment through a systems level deterministic and spatially homogeneous model to help characterize the hypersensitive response of the cancer cell lines harboring constitutively active mutant kinases to inhibitor treatment. By introducing a molecularly resolved branched network systems model (the molecular resolution is introduced for EGFR reactions and interactions in order to distinguish differences in activation between wild-type and mutants), we are able to quantify differences in (1) short-term signaling in downstream ERK and Akt activation, (2) the changes in the cellular inhibition EC50 associated with receptor phosphorylation (i.e., 50% inhibition of receptor phosphorylation in the cellular context), and (3) EC50 for the inhibition of activated downstream markers ERK-(p) and Akt-(p), where (p) denotes phosphorylated, upon treatment with the inhibitors in cell lines carrying both wild-type and mutant forms of the receptor. Using the branched signaling model, we illustrate a possible mechanism for preferential Akt activation in the cell lines harboring the oncogenic mutants of EGFR implicated in non-small-cell lung cancer and the enhanced efficacy of the inhibitor erlotinib especially in ablating the cellular Akt-(p) response. Using a simple phenomenological model to describe the effect of Akt activation on cellular decisions, we discuss how this preferential Akt activation is conducive to cellular oncogene addiction and how its disruption can lead to dramatic apoptotic response and hence remarkable inhibitor efficacies. We also identify key network nodes of our branched signaling model through sensitivity analysis as those rendering the network hypersensitive to enhanced ERK-(p) and Akt-(p); intriguingly, the identified nodes have a strong correlation with species implicated in oncogenic transformations in human cancers as well as in drug resistance mechanisms identified for the inhibitors in non-small-cell lung cancer therapy. PMID- 18412408 TI - Molecular dynamics simulation study on stabilities and reactivities of NADH cytochrome B5 reductase. AB - Binding free energies between coenzyme (FAD and NADH) and the apoenzyme of NADH cytochrome b5 reductase (b5R) were estimated by applying the continuum Poisson Boltzmann (PB) model to structures sampled from molecular dynamics simulations in explicit water molecules. Important residues for the enzymatic catalysis were clarified using a computational alanine scanning method. The binding free energies calculated by applying an alanine scanning method can successfully reproduce the trends of the measured steady-state enzymatic activities kcatNADH/KmNADH. Significant decreases in the binding free energy are expected when one of the four residues Arg91, Lys110, Ser127, and Thr181 is mutated into Ala. According to the results of the molecular dynamics simulation, Thr181 is considered to be one of the key residues that helps NADH to approach the isoalloxazine in FAD. Finally, we have constructed very simplified model systems and carried out density functional theory calculations using B3LYP/LANL2DZ//ROHF(or RHF)/LANL2DZ level of theory in order to elucidate a realistic and feasible mechanism of the hydride-ion transfer from NADH to FAD affected by HEME(Fe3+) as an electron acceptor. Our calculated results suggest that the electron and/or hydride-ion transfer reaction from NADH to FAD can be accelerated in the presence of HEME(Fe3+). PMID- 18412407 TI - Improved sampling for simulations of interfacial membrane proteins: application of generalized shadow hybrid Monte Carlo to a peptide toxin/bilayer system. AB - The computational costs associated with performing molecular dynamics (MD) simulations are still somewhat prohibitive and therefore limit the time and length scales that can be currently achieved. One approach to overcoming the limited size and duration of a simulation is to reduce the amount of detail when representing a system of interest, generally termed "coarse-graining". An alternative approach is via more efficient sampling methods that offer an enhanced search of a complex multidimensional energy landscape. One could also combine enhanced sampling methods with a coarse-grained (CG) force field. Here, we apply generalized shadow hybrid Monte Carlo (GSHMC), a recently proposed simulation protocol, to a biomolecular system of moderate size and show that GSHMC offers improved sampling compared to standard MD simulation. Our test system is a CG representation of a small peptide toxin interacting with a phospholipid bilayer. Specifically, we show that GSHMC allows for a quicker localization of the toxin to its equilibrium location of interaction at the headgroup/water interface of the bilayer. GSHMC therefore potentially allows for future exploration of larger and more complex systems over longer periods, which would otherwise be impractical to perform using conventional simulation methodology. PMID- 18412409 TI - Vibrational analysis of N-acetyl-alpha-D-glucosamine and beta-D-glucuronic acid. AB - Infrared spectra of solid and aqueous solutions of N-acetyl-alpha-D-glucosamine and beta-D-glucuronic acid have been investigated by means of Fourier transform infrared (FT-IR) spectroscopy and quantum chemical density functional theory (DFT) calculations. The errors of the computed harmonic force field were corrected according to the scaled quantum mechanical (SQM) method of Pulay, with scale factors partly from the literature and partly developed here. Scale factors for the hydrogen-bonded OH groups were determined by SQM treatment of ethylene glycol. The IR spectra and test computations revealed that beta-D-glucuronic acid is present as a dimer, formed by hydrogen-bonding between the COOH groups, in the solid phase. On the basis of the calculated results, 64 and 56 bands in the 4000 50 cm(-1) range of the FT-IR spectra have been assigned for N-acetyl-alpha-D glucosamine and beta-D-glucuronic acid, respectively. PMID- 18412410 TI - Quantum mechanically based estimation of perturbed-chain polar statistical associating fluid theory parameters for analyzing their physical significance and predicting properties. AB - We have computed molecular descriptors for sizes, shapes, charge distributions, and dispersion interactions for 67 compounds using quantum chemical ab initio and density functional theory methods. For the same compounds, we have fitted the three perturbed-chain polar statistical associating fluid theory (PCP-SAFT) equation of state (EOS) parameters to experimental data and have performed a statistical analysis for relations between the descriptors and the EOS parameters. On this basis, an analysis of the physical significance of the parameters, the limits of the present descriptors, and the PCP-SAFT EOS has been performed. The result is a method that can be used to estimate the vapor pressure curve including the normal boiling point, the liquid volume, the enthalpy of vaporization, the critical data, mixture properties, and so on. When only two of the three parameters are predicted and one is adjusted to experimental normal boiling point data, excellent predictions of all investigated pure compound and mixture properties are obtained. We are convinced that the methodology presented in this work will lead to new EOS applications as well as improved EOS models whose predictive performance is likely to surpass that of most present quantum chemically based, quantitative structure-property relationship, and group contribution methods for a broad range of chemical substances. PMID- 18412411 TI - Modeling membranes under a transmembrane potential. AB - Accurate modeling of ion transport through synthetic and biological transmembrane channels has been so far a challenging problem. We introduce here a new method that allows one to study such transport under realistic biological conditions. We present results from molecular dynamics simulations of an ion channel formed by a peptide nanotube, embedded in a lipid bilayer, and subject to transmembrane potentials generated by asymmetric distributions of ions on both sides of the membrane. We show that the method is efficient for generating ionic currents and allows us to estimate the intrinsic conductance of the channel. PMID- 18412412 TI - Electrochemical sensing of membrane potential and enzyme function using gallium arsenide electrodes functionalized with supported membranes. AB - We deposit phospholipid monolayers on highly doped p-GaAs electrodes that are precoated with methyl-mercaptobiphenyl monolayers and operate such a biofunctional electrolyte-insulator-semiconductor (EIS) setup as an analogue of a metal-oxide-semiconductor setup. Electrochemical impedance spectra measured over a wide frequency range demonstrate that the presence of a lipid monolayer remarkably slows down the diffusion of ions so that the membrane-functionalized GaAs can be subjected to electrochemical investigations for more than 3 days with no sign of degradation. The biofunctional EIS setup enables us to translate changes in the surface charge density Q and bias potentials Ubias into the change in the interface capacitance Cp. Since Cp is governed by the capacitance of semiconductor space charge region CSC, the linear relationships obtained for 1/Cp2 vs Q and 1/Cp2 vs Ubias suggests that Cp can be used to detect the surface charges with a high sensitivity (1 charge per 18 nm2). Furthermore, the kinetics of phospholipids degradation by phospholipase A2 can also be monitored by a significant decrease in diffusion coefficients through the membrane by a factor of 104. Thus, the operation of GaAs membrane composites established here allows for electrochemical sensing of surface potential and barrier capability of biological membranes in a quantitative manner. PMID- 18412413 TI - Parametrization, molecular dynamics simulation, and calculation of electron spin resonance spectra of a nitroxide spin label on a polyalanine alpha-helix. AB - The nitroxide spin label 1-oxyl-2,2,5,5-tetramethylpyrroline-3-methyl methanethiosulfonate (MTSSL), commonly used in site-directed spin labeling of proteins, is studied with molecular dynamics (MD) simulations. After developing force field parameters for the nitroxide moiety and the spin label linker, we simulate MTSSL attached to a polyalanine alpha-helix in explicit solvent to elucidate the factors affecting its conformational dynamics. Electron spin resonance spectra at 9 and 250 GHz are simulated in the time domain using the MD trajectories and including global rotational diffusion appropriate for the tumbling of T4 Lysozyme in solution. Analysis of the MD simulations reveals the presence of significant hydrophobic interactions of the spin label with the alanine side chains. PMID- 18412414 TI - Accelerating QM/MM free energy calculations: representing the surroundings by an updated mean charge distribution. AB - Reliable studies of enzymatic reactions by combined quantum mechanical/molecular mechanics (QM(ai)/MM) approaches with an ab initio description of the quantum region presents a major challenge to computational chemists. The main problem is the need for very large computer time to evaluate the QM energy, which in turn makes it extremely challenging to perform proper configurational sampling. One of the most obvious options for accelerating QM/MM simulations is the use of an average solvent potential. In fact, the idea of using an average solvent potential is rather obvious and has implicitly been used in Langevin dipole/QM calculations. However, in the case of explicit solvent models the practical implementations are more challenging, and the accuracy of the averaging approach has not been validated. The present study introduces the average effect of the fluctuating solvent charges by using equivalent charge distributions, which are updated every m steps. Several models are evaluated in terms of the resulting accuracy and efficiency. The most effective model divides the system into an inner region with N explicit solvent atoms and an external region with two effective charges. Different models are considered in terms of the division of the solvent system and the update frequency. Another key element of our approach is the use of the free energy perturbation (FEP) and/or linear response approximation treatments that guarantees the evaluation of the rigorous solvation free energy. Special attention is paid to the convergence of the calculated solvation free energies and the corresponding solute polarization. The performance of the method is examined by evaluating the solvation of a water molecule and a formate ion in water and also the dipole moment of water in water solution. Remarkably, it is found that different averaging procedures eventually converge to the same value but some protocols provide optimal ways of obtaining the final QM(ai)/MM converged results. The current method can provide computational time saving of 1000 for properly converging simulations relative to calculations that evaluate the QM(ai)/MM energy every time step. A specialized version of our approach that starts with a classical FEP charging and then evaluates the free energy of moving from the classical potential to the QM/MM potential appears to be particularly effective. This approach should provide a very powerful tool for QM(ai)/MM evaluation of solvation free energies in aqueous solutions and proteins. PMID- 18412415 TI - Standard partial molar volumes of some aqueous alkanolamines and alkoxyamines at temperatures up to 325 degrees C: functional group additivity in polar organic solutes under hydrothermal conditions. AB - Apparent molar volumes of dilute aqueous solutions of monoethanolamine (MEA), diethanolamine (DEA), triethanolamine (TEA), N,N-dimethylethanolamine (DMEA), ethylethanolamine (EAE), 2-diethylethanolamine (2-DEEA), and 3-methoxypropylamine (3-MPA) and their salts were measured at temperatures from 150 to 325 degrees C and pressures as high as 15 MPa. The results were corrected for the ionization and used to obtain the standard partial molar volumes, Vo2. A three-parameter equation of state was used to describe the temperature and pressure dependence of the standard partial molar volumes. The fitting parameters were successfully divided into functional group contributions at all temperatures to obtain the standard partial molar volume contributions. Including literature results for alcohols, carboxylic acids, and hydroxycarboxylic acids yielded the standard partial molar volume contributions of the functional groups >CH-, >CH2, -CH3, OH, -COOH, -O-, -->N, >NH, -NH2, -COO-Na+, -NH3+Cl-, >NH2+Cl-, and -->NH+Cl- over the range (150 degrees C d excitation energies of atoms, atomic ionization energies, and the dissociation energies of the diatomic hydrides. The Brueckner orbital-based BD(T) method performs substantially better than Hartree-Fock-based CCSD(T) for molecules such as NiH, where the starting unrestricted Hartree-Fock wavefunction suffers from a high degree of spin contamination. Comparison with available data for geometries of transition metal hydrides also shows good agreement. A smaller basis set without core polarization functions, G3MP2Large, is also defined. PMID- 18412439 TI - Computed lifetimes of metastable states of the NO2+ dication. AB - Based on the ab initio potential energy, spin-orbit coupling, electronic transition dipole moment, and radial nonadiabatic coupling functions, the energy level positions, lifetimes, and radiative transition probabilities (Einstein A coefficients) have been determined for the lowest electronic states of NO2+ using the log-amplitude-phase, stabilization, and complex-scaling methods. The calculated characteristics are in reasonable agreement to the available experimental data, thus, evidencing the reliability of the theoretical predictions for the characteristics unobserved to date. With the exception of the vKF+H reaction and comparison with quasiclassical trajectory results. AB - Extensive quantum real wave packet calculations within the helicity decoupling approximation are used to analyze the influence of the HF vibrational excitation on the K+HF(v=0-2,j=0)-->KF+H reaction. Quantum reaction probabilities P and reaction cross sections sigma are compared with corresponding quasiclassical trajectory (QCT) results. Disregarding threshold regions for v=0 and 1 (v=2 has no threshold), both approaches lead to remarkably similar results, particularly for sigma, validating the use of the QCT method for this system. When moving from v=0 to v=1 there is a large increase in P and sigma, as expected for a late barrier system. For v=2 the reaction becomes exoergic and P approximately 0.95 (with the exception of large total angular momenta where centrifugal barriers play a role). While substantial vibrational enhancement of the reactivity is thus seen, it is still quite less than that inferred from experimental data in the intermediate and high collision energy ranges. The origin of this discrepancy is unclear. PMID- 18412441 TI - Excitation levels and magic numbers of small parahydrogen clusters (NSF+ +F that relates to the REMPI spectrum was discussed. PMID- 18412445 TI - Structures of MAu16 (-) (M=Ag, Li, Na, and K): how far is the endohedral doping? AB - The structural and electronic properties of MAu16 (-) (M=Ag, Li, Na, and K) have been studied by the scalar relativistic all-electron density-functional calculations, in which particular attention is paid to the stability of the endohedral Au16 (-) cage doped by different dopant atoms. It is found that only the smaller atoms, such as Cu, Li, and Na, can be stably encapsulated in the Au16 (-) cage, while the addition of the larger Ag or K atom prefers to locate in the surface or outside of the cage, which is inconsistent with the previous hypothesis that the Au16 (-) cage could act as a container to hold an arbitrary heterometal atom. The stable endohedral Li@Au16 (-) and Na@Au16 (-) have a large energy gap between the highest occupied molecular orbital and the lowest unoccupied molecular orbital gap, indicating that they are chemically stable and may be used as potential building blocks for designing cluster-assembled materials. PMID- 18412446 TI - Ab initio investigation of the electronic structure and bonding of BH, BH(-), and HBBH molecules. AB - By correlating all electrons and employing core-tuned correlation consistent basis sets of quintuple-zeta quality, we applied multireference and coupled cluster methods to study 32 electronic states of the diatomic BH molecule, two bound states of BH(-), and three states of the linear HBBH molecule. We have constructed full potential energy curves and profiles, reporting binding energies, geometries, spectroscopic parameters, dipole moments, and energy separations, whereas our numerical results are in excellent agreement with available experimental numbers. We are trying as well to interpret the binding modes of a large number of the examined states. 18 states of BH are of Rydberg character, with the BH(-) anion revealing similar structural characteristics to the isoelectronic CH species. The first three states of HBBH X 3Sigma g (-), a 1Delta g, and b 1Sigma g + diabatically correlate to two a 3Pi BH fragments, they are similar to the states b 3Sigma g (-), B 1Delta g, and B' 1Sigma g + of the isoelectronic molecule C2, however, their ordering follows that of the first three states of the O2 molecule. PMID- 18412447 TI - An ab initio study based on a finite nucleus model for isotope fractionation in the U(III)-U(IV) exchange reaction system. AB - Isotope fractionation in the U(III)-U(IV) reaction system was investigated by a series of atomic relativistic ab initio calculations using the multiconfigurational Dirac-Coulomb Hartree-Fock method. To evaluate the nuclear volume effect on the fractionation, the Fermi statistical distribution function was adopted for nuclear charge density. The isotope fractionation coefficient epsilon resulting from the nuclear volume difference was evaluated from the total electronic energies of U3+ and U4+, based on the theoretical equation proposed by Bigeleisen [J. Am. Chem. Soc. 118, 3676 (1996)]. The calculated fractionation coefficient epsilon in the present work for the isotopic pair 235U and 238U at 293 K is 0.0031, which is quite close to the experimentally observed value of 0.0027. Discussion is extended to the nuclear volume effects on isotopic fractionations in the Pu(III)-Pu(IV) and Eu(II)-Eu(III) exchange systems. PMID- 18412448 TI - The role of the excited electronic states in the C+ + H2O reaction. AB - The electronic excited states of the [COH2]+ system have been studied in order to establish their role in the dynamics of the C+ + H2O-->[COH]+ +H reaction, which is a prototypical ion-molecule reaction. The most relevant minima and saddle points of the lowest excited state have been determined and energy profiles for the lowest excited doublet and quartet electronic states have been computed along the fragmentation and isomerization coordinates. Also, nonadiabatic coupling strengths between the ground and the first excited state have been computed where they can be large. Our analysis suggests that the first excited state could play an important role in the generation of the formyl isomer, which has been detected in crossed beam experiments [D. M. Sonnenfroh et al., J. Chem. Phys. 83, 3985 (1985)], but could not be explained in quasiclassical trajectory computations [Y. Ishikawa et al., Chem. Phys. Lett. 370, 490 (2003); J. R. Flores, J. Chem. Phys. 125, 164309 (2006)]. PMID- 18412449 TI - Recoil excitation of vibrational structure in the carbon 1s photoelectron spectrum of CF4. AB - The carbon 1s photoelectron spectrum of CF4 measured at photon energies from 330 to 1500 eV shows significant contributions from nonsymmetric vibrational modes. These increase linearly as the photon energy increases. The excitation of these modes, which is not predicted in the usual Franck-Condon point of view, arises from the recoil momentum imparted to the carbon atom in the ionization process. A theory is presented for quantitative prediction of the recoil effect; the predictions of this theory are in agreement to the measurements. The experiments also yield the vibrational frequencies of the symmetric and asymmetric stretching modes in core-ionized CF4, the change in CF bond length upon ionization, -0.61 pm, and the Lorentzian linewidth of the carbon 1s hole, 67 meV. PMID- 18412450 TI - The permanent electric dipole moment of calcium monodeuteride. AB - The sub-Doppler laser induced fluorescence spectra of numerous branch features in the B 2Sigma+ -X 2Sigma+(0,0) band of calcium monodeuteride were recorded field free and in the presence of a static electric field of up to 7 kV/cm. The field free spectra were analyzed to produce an improved set of fine structure parameters for the B 2Sigma+(v=0) state. The observed electric field induced splittings and shifts were analyzed to produce permanent electric dipole moments of 2.57(3) and 2.51(3) D for B 2Sigma+(v=0) and X 2Sigma+(v=0) states, respectively. The permanent electric dipole moment for the X 2Sigma+(v=0) state of CaH is estimated to be 2.53(3) D. PMID- 18412451 TI - The photodissociation of the water dimer in the A band: a twelve-dimensional quasiclassical study. AB - The quasiclassical absorption spectrum of the water dimer in the A band was calculated taking into account motion in all degrees of freedom of the system. The ab initio excited state potentials employed were interpolated by the modified Shepard interpolation method using QMRCI energies and state-averaged MCSCF gradients and Hessians. The ground state vibrational wavefunction was variationally calculated using an adiabatic separation between the high and low frequency normal modes of the system. The calculated spectrum of water dimer shows a clear blueshift with respect to the monomer, but also a small red tail, in agreement with the prediction by Harvey et al. [J. Chem. Phys. 109, 8747 (1998)]. Previous three-dimensional model studies of the photodissociation of the water dimer by Valenzano et al. [J. Chem. Phys. 123, 034303 (2005)] did not show this red tail. A thorough analysis of the dependence of the spectrum on the modes coupled explicitly in the calculation of the spectrum shows that the red tail is due to coupling between the intramolecular stretch vibrations on different monomers. PMID- 18412452 TI - Prediction of metastable metal-rare gas fluorides: FMRgF (M=Be and Mg; Rg=Ar, Kr and Xe). AB - The structure, stability, charge redistribution, bonding, and harmonic vibrational frequencies of rare gas containing group II-A fluorides with the general formula FMRgF (where M=Be and Mg; Rg=Ar, Kr, and Xe) have been investigated using second order Moller-Plesset perturbation theory, density functional theory, and coupled cluster theory [CCSD(T)] methods. The species, FMRgF show a quasilinear structure at the minima and a bent structure at the transition state. The predicted species are unstable with respect to the two-body dissociation channel, leading to the global minima (MF2+Rg) on the singlet potential energy surface. However, with respect to other two-body dissociation channel (FM+RgF), they are found to be stable and have high positive energies on the same surface. The computed binding energy for the two-body dissociation channels are 94.0, 164.7, and 199.7 kJ mol(-1) for FBeArF, FBeKrF, FBeXeF, respectively, at CCSD(T) method. The corresponding energy values are 83.4, 130.7, and 180.1 kJ mol(-1) for FMgArF, FMgKrF, and FMgXeF, respectively, at the same level of theory. With respect to the three-body dissociation (FM+Rg+F) channel as well as dissociation into atomic constituent, they are also found to be stable and have high positive energies. The dissociation of the predicted species typically proceeds via MRgF bending mode at the transition state. The computed barrier heights for the transition states are 11.4, 32.2, and 57.6 kJ mol(-1) for FBeArF, FBeKrF, and FBeXeF, respectively, at the CCSD(T) method. The corresponding barrier heights for the Mg containing species are 2.1, 9.2, and 32.1 kJ mol(-1) along the series Ar--Kr--Xe, respectively. The M--Rg bond energies of the FMRgF species is significantly higher than the corresponding bond energies of the M+--Rg species ( approximately 53 and approximately 15 kJ mol(-1) for Be+--Ar and Mg+--Ar, respectively). The computed energy diagram as well as the geometrical parameters along with the AIM results suggest that the species are metastable with partial covalent character in the M--Rg bonding. Thus, it may be possible to prepare and to characterize these species using low temperature matrix isolation technique. PMID- 18412453 TI - A hybrid explicit/implicit solvation method for first-principle molecular dynamics simulations. AB - In this work, we present a hybrid explicit/implicit solvation model, well suited for first-principles molecular dynamics simulations of solute-solvent systems. An effective procedure is presented that allows to reliably model a solute with a few explicit solvation shells, ensuring solvent bulk behavior at the boundary with the continuum. Such an approach is integrated with high-level ab initio methods using localized basis functions to perform first-principles or mixed quantum mechanics/molecular mechanics simulations within the extended-Lagrangian formalism. A careful validation of the model along with illustrative applications to solutions of acetone and glycine radical are presented, considering two solvents of different polarity, namely, water and chloroform. Results show that the present model describes dynamical and solvent effects with an accuracy at least comparable to that of conventional approaches based on periodic boundary conditions. PMID- 18412454 TI - Proton transfer in a polar solvent from ring polymer reaction rate theory. AB - We have used the ring polymer molecular dynamics method to study the Azzouz Borgis model for proton transfer between phenol (AH) and trimethylamine (B) in liquid methyl chloride. When the A-H distance is used as the reaction coordinate, the ring polymer trajectories are found to exhibit multiple recrossings of the transition state dividing surface and to give a rate coefficient that is smaller than the quantum transition state theory value by an order of magnitude. This is to be expected on kinematic grounds for a heavy-light-heavy reaction when the light atom transfer coordinate is used as the reaction coordinate, and it clearly precludes the use of transition state theory with this reaction coordinate. As has been shown previously for this problem, a solvent polarization coordinate defined in terms of the expectation value of the proton transfer distance in the ground adiabatic quantum state provides a better reaction coordinate with less recrossing. These results are discussed in light of the wide body of earlier theoretical work on the Azzouz-Borgis model and the considerable range of previously reported values for its proton and deuteron transfer rate coefficients. PMID- 18412455 TI - Nature of metal-nonmetal transition in metal-ammonia solutions. II. From uniform metallic state to inhomogeneous electronic microstructure. AB - Applying semianalytical models of nonideal plasma, we evaluate the behavior of the metallic phase in metal-ammonia solutions (MAS). This behavior is mainly controlled by the degenerate electron gas, which remains stable down to 5 MPM due to high solvent polarizability and strong dielectric screening of solvated ions. Comparing the behavior of the metallic state with those of localized solvated electrons, we have estimated the miscibility gap Delta n for various alkali metals and found Delta n(Na)>Delta n(K). It is rather narrow in Rb-NH3 and does not occur in Cs-NH3 solutions, which is in full agreement with the experiments. The case of Li is discussed separately. The difference calculated in the excess free energies of the metallic and nonmetallic phases is in the order of kBT, yielding a thermally fluctuating mixed state at intermediate metal concentrations. It results in a continuous metal-nonmetal (MNM) transition above the consolute point Tc and a phase separation below Tc. We propose a criterion for the MNM transition which may be attributed to the line of the maximum of compressibility above Tc. This line crosses the spinodal one at the critical temperature. Finally, we assert that a new electronic phase similar to microemulsion should also arise between the spinodal and the binodal lines. PMID- 18412456 TI - Theoretical and numerical study of the phase diagram of patchy colloids: ordered and disordered patch arrangements. AB - We report theoretical and numerical evaluations of the phase diagram for a model of patchy particles. Specifically, we study hard spheres whose surface is decorated by a small number f of identical sites ("sticky spots") interacting via a short-ranged square-well attraction. We theoretically evaluate, solving the Wertheim theory, the location of the critical point and the gas-liquid coexistence line for several values of f and compare them to the results of Gibbs and grand canonical Monte Carlo simulations. We study both ordered and disordered arrangements of the sites on the hard-sphere surface and confirm that patchiness has a strong effect on the phase diagram: the gas-liquid coexistence region in the temperature-density plane is significantly reduced as f decreases. We also theoretically evaluate the locus of specific heat maxima and the percolation line. PMID- 18412457 TI - Thermodynamics and dynamics of a monoatomic glass former. Constant pressure and constant volume behavior. AB - We report constant-volume and constant-pressure simulations of the thermodynamic and dynamic properties of the low-temperature liquid and crystalline phases of the modified Stillinger-Weber (SW) model. We have found an approximately linear temperature increase of the effective Gaussian width of the distribution of inherent structures. This effect comes from non-Gaussianity of the landscape and is consistent with the predictions of the Gaussian excitations model representing the thermodynamics of the configurational manifold as an ensemble of excitations, each carrying an excitation entropy. The SW model provides us with both the configurational and excess entropies, with the difference mostly attributed to vibrational anharmonicity. We therefore can address the distinction between the excess thermodynamic quantities, often used to interpret experiments, and configurational thermodynamics used to describe the dynamics in the Adam-Gibbs (AG) equation. However we are limited computationally to work at temperatures above the "crossover" temperature at which the breakdown in the Adam-Gibbs relation has been identified in laboratory studies. We find a new break in the slope of the constant pressure AG plot (in the same sense but at much higher temperature than with laboratory data) when the excess entropy is used in the AG equation. This break, which we associate with anharmonic vibrational effects, is not seen when the configurational entropy is used. The simulation diffusivity data are equally well fitted by the AG equation and by a new equation, derived within the Gaussian excitations model, that emphasizes enthalpy over entropy as the thermodynamic control variable for transport in viscous liquids. We show that the modified SW model has close links to the behavior observed for bulk metallic glasses, both in its diffusional and in its thermodynamic properties. PMID- 18412458 TI - Optically controlled resonance energy transfer: mechanism and configuration for all-optical switching. AB - In a molecular system of energy donors and acceptors, resonance energy transfer is the primary mechanism by means of which electronic energy is redistributed between molecules, following the excitation of a donor. Given a suitable geometric configuration it is possible to completely inhibit this energy transfer in such a way that it can only be activated by application of an off-resonant laser beam: this is the principle of optically controlled resonance energy transfer, the basis for an all-optical switch. This paper begins with an investigation of optically controlled energy transfer between a single donor and acceptor molecule, identifying the symmetry and structural constraints and analyzing in detail the dependence on molecular energy level positioning. Spatially correlated donor and acceptor arrays with linear, square, and hexagonally structured arrangements are then assessed as potential configurations for all-optical switching. Built on quantum electrodynamical principles the concept of transfer fidelity, a parameter quantifying the efficiency of energy transportation, is introduced and defined. Results are explored by employing numerical simulations and graphical analysis. Finally, a discussion focuses on the advantages of such energy transfer based processes over all-optical switching of other proposed forms. PMID- 18412459 TI - Vibrational dynamics and structural investigation of 2,2'-dipyridylketone using Raman, IR and UV-visible spectroscopy aided by ab initio and density functional theory calculation. AB - Detailed investigation on the vibrational and electronic spectra has been carried out in order to study various properties of 2,2'-dipyridylketone molecule in its ground and excited electronic states. To get insight into the structural and symmetry features of the molecule, Raman excitation profiles of several normal modes have been analyzed. The polarized Raman spectra in different environments along with their IR counterpart have been critically surveyed and different normal modes have been assigned. The knowledge in regard to the positions of different excited electronic states has been acquired from the study of electronic absorption spectra. All the experimental observations have been substantiated and corroborated theoretically by the quantum chemical calculation. Possibility of exciton splitting of the 1La band has been explored both from theoretical and experimental points of view. PMID- 18412460 TI - A mode-coupling theory treatment of the transport coefficients of the Lennard Jones fluid. AB - We apply mode-coupling theory to study shear viscosity and self-diffusion coefficient of the Lennard-Jones fluid throughout the entire fluid region of the phase diagram. Theoretical results are compared with the extensive simulation data and good agreement is found. In addition, theory is compared to the experimental data on the transport coefficients of inert gas fluids. PMID- 18412461 TI - Scaling law of stable single cells in density fingering of chemical fronts. AB - Stable single fingers evolving in a thin layer of hydrodynamically unstable reactive solution are investigated in moderately wide Hele-Shaw cells. The mixing length associated with the final pattern is shown to be proportional to the width of the container. The scaling law, which is best illustrated by the overlay of temporal average front profiles, is demonstrated for two chemical compositions and boundaries of different thermal conductances. The observed single finger is found to be independent of the orientation of the reaction vessel for a wide range of tilt angles. PMID- 18412462 TI - Polarized pump-probe measurements of electronic motion via a conical intersection. AB - Polarized femtosecond pump-probe spectroscopy is used to observe electronic wavepacket motion for vibrational wavepackets centered on a conical intersection. After excitation of a doubly degenerate electronic state in a square symmetric silicon naphthalocyanine molecule, electronic motions cause a approximately 100 fs drop in the polarization anisotropy that can be quantitatively predicted from vibrational quantum beat modulations of the pump-probe signal. Vibrational symmetries are determined from the polarization anisotropy of the vibrational quantum beats. The polarization anisotropy of the totally symmetric vibrational quantum beats shows that the electronic wavepackets equilibrate via the conical intersection within approximately 200 fs. The relationship used to predict the initial electronic polarization anisotropy decay from the asymmetric vibrational quantum beat amplitudes indicates that the initial width of the vibrational wavepacket determines the initial speed of electronic wavepacket motion. For chemically reactive conical intersections, which can have 1000 times greater stabilization energies than the one observed here, the same theory predicts electronic equilibration within 2 fs. Such electronic movements would be the fastest known chemical processes. PMID- 18412463 TI - Test of the consistency of various linearized semiclassical initial value time correlation functions in application to inelastic neutron scattering from liquid para-hydrogen. AB - The linearized approximation to the semiclassical initial value representation (LSC-IVR) is used to calculate time correlation functions relevant to the incoherent dynamic structure factor for inelastic neutron scattering from liquid para-hydrogen at 14 K. Various time correlations functions were used which, if evaluated exactly, would give identical results, but they do not because the LSC IVR is approximate. Some of the correlation functions involve only linear operators, and others involve nonlinear operators. The consistency of the results obtained with the various time correlation functions thus provides a useful test of the accuracy of the LSC-IVR approximation and its ability to treat correlation functions involving both linear and nonlinear operators in realistic anharmonic systems. The good agreement of the results obtained from different correlation functions, their excellent behavior in the spectral moment tests based on the exact moment constraints, and their semiquantitative agreement with the inelastic neutron scattering experimental data all suggest that the LSC-IVR is indeed a good short-time approximation for quantum mechanical correlation functions. PMID- 18412464 TI - Thermal history effects and methyl tunneling dynamics in a supramolecular complex of calixarene and para-xylene. AB - The low-temperature structure and dynamics of guest molecules of p-xylene incorporated in the isopropyl-calix[4] arene(2:1) p-xylene complex have been investigated by solid state nuclear magnetic resonance (NMR). Using one dimensional 1H-decoupled 13C cross-polarization magic-angle-spinning (MAS) NMR and two-dimensional 1H-13C correlation spectroscopy, a full assignment of the 13C and 1H chemical shifts has been made. Using 1H NMR relaxometry, the effects of thermal history on the structure of the system have been investigated. Rapidly cooled samples have 1H spin-lattice relaxation times T1, which at low temperature (T<60 K) are typically two orders of magnitude faster than those observed in annealed samples which have been cooled slowly over many hours. In both forms, the low-temperature relaxation is driven by the dynamics of the weakly hindered methyl rotors of the p-xylene guest. The substantial difference in T1 is attributed in the rapidly cooled sample to disorder in the structure of the complex leading to a wide distribution of correlation times and methyl barrier heights. A comparison of the linewidths and splittings in the high resolution 13C MAS spectra of the two forms provides structural insight into the nature of the disorder. Using 1H field-cycling NMR relaxometry, the methyl dynamics of the p xylene guest in the annealed sample have been fully characterized. The B-field dependence of the 1H T1 maps out the spectral density from which the correlation times are directly measured. The methyl barrier heights are determined from an analysis of the temperature dependence. PMID- 18412465 TI - Combining density functional theory and cluster expansion methods to predict H2 permeance through Pd-based binary alloy membranes. AB - First-principles calculations offer a useful complement to experimental approaches for characterizing hydrogen permeance through dense metal membranes. A challenge in applying these methods to disordered alloys is to make quantitative predictions for the net solubility and diffusivity of interstitial H based on the spatially local information that can be obtained from first-principles calculations. In this study, we used a combination of density functional theory calculations and a cluster expansion method to describe interstitial H in alloys of composition Pd96M4, where M=Ag, Cu, and Rh. The cluster expansion approach highlights the shortcomings of simple lattice models that have been used in the past to study similar systems. We use Sieverts' law to calculate H solubility and a kinetic Monte Carlo scheme to find the diffusivity of H in PdAg, PdCu, and PdRh alloys at a temperature range of 400neutral>cation, but the values are rather similar for all three charge states. The thermodynamic stability for the loss of GaAs is reported. PMID- 18412472 TI - Pressure effects on CrCl6(3-) embedded in cubic Cs2NaMCl6 (M=Sc,Y) lattices: study through periodic and cluster calculations. AB - The structural, elastic, vibrational, and optical properties of cubic elpasolites Cs2NaMCl6 (M=Sc,Y) containing CrCl6(3-) complexes have been investigated by means of both periodic and cluster calculations as a function of pressure in the framework of density functional theory. Aside from calculating the host lattice bulk modulus BH and the local modulus B1 associated with the CrCl6(3-), complex particular attention is paid to the pressure dependence of Huang-Rhys factors, Sa and Se (related to local a1g and eg modes), and the Stokes shift associated with the first electronic excited state 4T2g (t2g 2eg) of CrCl6(3-). The present calculations provide a big difference between BH=231 kbars and B1=676 kbars derived for Cs2NaScCl6:Cr3+ at zero pressure which plays a key role for a right interpretation of pressure effects on vibration frequencies and optical parameters due to CrCl6(3-). The significant decrease of Huang-Rhys factors, Sa and Se, due to the pressure observed experimentally is well accounted for by the present work which supports that partial differential Sa/ partial differential P is determined by the Gruneisen constant gamma a of the a1g local mode (whose frequency is nu a) and the dependence of 10Dq on the metal-ligand distance. At the same time, the present results point out that the Stokes shift would be little pressure dependent in the range of 0-50 kbars. Accordingly the Ham effect in the 4T2g (t2g 2eg) state of CrCl6(3-) in the cubic elpasolites would also happen for a pressure up to 50 kbars but the spin-orbit constant would increase with respect to that at zero pressure. From the analysis carried out in this work it is also concluded that the figures d nu a/dP=0.55 cm(-1)kbar and dSa/dP=-7.2 x 10(-3) kbar(-1) extracted from the complex emission band of Cs2NaScCl6:Cr3+ are hardly compatible. This fact underlines the usefulness of ab initio calculations for helping in the analysis of complex experimental findings. Finally, as the CrCl6(3-) unit is found to be to a good extent elastically decoupled from the rest of the elpasolite lattice, a model is shown to lead to an approximate relation between the pressure derivative of the local modulus and the Gruneisen constant gamma a. PMID- 18412473 TI - Characterization of enantiospecific chemisorption on chiral Cu surfaces vicinal to Cu(111) and Cu(100) using density functional theory. AB - Surfaces of simple fcc metals such as Cu with nonzero and unequal Miller indices are intrinsically chiral. Density functional theory (DFT) calculations are a useful way to study the enantiospecific adsorption of small chiral molecules on these chiral metal surfaces. We report DFT calculations of seven chiral molecules on several structurally distinct chiral Cu surfaces. These surfaces include two surfaces with (111)-oriented terraces and one with (100)-oriented terraces. Calculations are also described on a surface that was modified to mimic the surface structures that typically appear on real metal surfaces following thermally driven fluctuations in step edges. Our results provide initial information on how variation in the surface structure of intrinsically chiral metal surfaces can affect the enantiospecific adsorption of small molecules on these surfaces. PMID- 18412474 TI - Cage and tube structures of medium-sized zinc oxide clusters (ZnO)n (n=24, 28, 36, and 48). AB - Following our recent work which revealed that the lowest-energy structures of (ZnO)n (n=9-18) follow cage and tube structural growth patterns with stacks of small subunits of (ZnO)2 and (ZnO)3 [Wang et al., J. Phys. Chem. C 111, 4956 (2007)], we have extended the search for the most stable structures to some larger clusters, i.e., (ZnO)n (n=24, 28, 36, and 48) by using gradient-corrected density-functional theory (DFT). A number of starting configurations belonging to different structural motifs were generated from handmade constructions with chemical intuition and then optimized via DFT calculations. Within the size range studied, cage and tube structures were found to be the most preferred structural motifs for the (ZnO)n clusters. PMID- 18412475 TI - Extracting maximum information from polarized surface vibrational spectra: application to etched, H-terminated Si(110) surfaces. AB - A general method to maximize the information extracted from polarized surface absorption spectra is developed and applied to the study of etched Si(110) surfaces. In essence, this technique transforms spectra from the experimental reference frame, which is defined by the direction of the surface electric field during irradiation by s- and p-polarized light, into a more appropriate Cartesian reference frame defined by the surface normal and the plane of incidence. If the Cartesian reference frame is aligned with high symmetry directions of the system, significant spectral simplification can result. This analysis relies on the well known boundary conditions on interfacial electric fields and is independent of any adsorbate screening or the effective dielectric constant of the adsorbate layer. The validity of this analysis is demonstrated on the spectra of NH4F etched, H-terminated Si(110). The transition dipole moments of the symmetric and antisymmetric Si[Single Bond]H stretch modes associated with flat terraces are polarized along the [110] and [001] directions, respectively. Two additional modes with transition dipoles polarized along the [001] and [110] directions are assigned to defect species associated with microfaceting and other surface roughness. Data taken in two different experimental geometries are shown to be in excellent quantitative agreement, confirming the validity of the technique. Additionally, the measured adsorbate layer dielectric constant is in good agreement with previously reported values for hydrogen-terminated silicon surfaces. PMID- 18412476 TI - Conformational behavior of polymers adsorbed on nanotubes. AB - The importance of hydrophobic interactions in determining polymer adsorption and wrapping of carbon nanotubes is still under debate. In this work, we concentrate on the effect of short-ranged weakly attractive hydrophobic interactions between polymers and nanotubes (modeled as an infinitely long and smooth cylindrical surface), neglecting all other interactions apart for chain flexibility. Using coarse-grained Monte Carlo simulation of such simplified systems, we find that uniform adsorption and wrapping of the nanotube occur for all degrees of chain flexibility for tubes with sufficiently large outer radii. However, the adsorbed conformations depend on chain stiffness, ranging from randomly adsorbed conformations of the flexible chain to perfect helical or multihelical conformations (in the case of more concentrated solutions) of the rigid chains. Adsorption appears to occur in a sequential manner, wrapping the nanotube nearly one monomer at a time from the point of contact. Once adsorbed, the chains travel on the surface of the cylinder, retaining their helical conformations for the semiflexible and rigid chains. Our findings may provide additional insight to experimentally observed ordered polymer wrapping of carbon nanotubes. PMID- 18412477 TI - Multiparticle collision dynamics modeling of viscoelastic fluids. AB - In order to investigate the rheological properties of viscoelastic fluids by mesoscopic hydrodynamics methods, we develop a multiparticle collision (MPC) dynamics model for a fluid of harmonic dumbbells. The algorithm consists of alternating streaming and collision steps. The advantage of the harmonic interactions is that the integration of the equations of motion in the streaming step can be performed analytically. Therefore, the algorithm is computationally as efficient as the original MPC algorithm for Newtonian fluids. The collision step is the same as in the original MPC method. All particles are confined between two solid walls moving oppositely, so that both steady and oscillatory shear flows can be investigated. Attractive wall potentials are applied to obtain a nearly uniform density everywhere in the simulation box. We find that both in steady and oscillatory shear flows, a boundary layer develops near the wall, with a higher velocity gradient than in the bulk. The thickness of this layer is proportional to the average dumbbell size. We determine the zero-shear viscosities as a function of the spring constant of the dumbbells and the mean free path. For very high shear rates, a very weak "shear thickening" behavior is observed. Moreover, storage and loss moduli are calculated in oscillatory shear, which show that the viscoelastic properties at low and moderate frequencies are consistent with a Maxwell fluid behavior. We compare our results with a kinetic theory of dumbbells in solution, and generally find good agreement. PMID- 18412478 TI - Dissipative particle dynamics simulation of depletion layer and polymer migration in micro- and nanochannels for dilute polymer solutions. AB - The flows of dilute polymer solutions in micro- and nanoscale channels are of both fundamental and practical importance in variety of applications in which the channel gap is of the same order as the size of the suspended particles or macromolecules. In such systems depletion layers are observed near solid-fluid interfaces, even in equilibrium, and the imposition of flow results in further cross-stream migration of the particles. In this work we employ dissipative particle dynamics to study depletion and migration in dilute polymer solutions in channels several times larger than the radius of gyration (Rg) of bead-spring chains. We compare depletion layers for different chain models and levels of chain representation, solvent quality, and relative wall-solvent-polymer interactions. By suitable scaling the simulated depletion layers compare well with the asymptotic lattice theory solution of depletion near a repulsive wall. In Poiseuille flow, polymer migration across the streamlines increases with the Peclet and the Reynolds number until the center-of-mass distribution develops two symmetric off-center peaks which identify the preferred chain positions across the channel. These appear to be governed by the balance of wall-chain repulsive interactions and an off-center driving force of the type known as the Segre Silberberg effect. PMID- 18412479 TI - Normal mode partitioning of Langevin dynamics for biomolecules. AB - We propose a novel normal mode multiple time stepping Langevin dynamics integrator called NML. The aim is to approximate the kinetics or thermodynamics of a biomolecule by a reduced model based on a normal mode decomposition of the dynamical space. Our basis set uses the eigenvectors of a mass reweighted Hessian matrix calculated with a biomolecular force field. This particular choice has the advantage of an ordering according to the eigenvalues, which have a physical meaning of being the square of the mode frequency. Low frequency eigenvalues correspond to more collective motions, whereas the highest frequency eigenvalues are the limiting factor for the stability of the integrator. In NML, the higher frequency modes are overdamped and relaxed near their energy minimum while respecting the subspace of low frequency dynamical modes. Our numerical results confirm that both sampling and rates are conserved for an implicitly solvated alanine dipeptide model, with only 30% of the modes propagated, when compared to the full model. For implicitly solvated systems, NML gives a twofold improvement in efficiency over plain Langevin dynamics for sampling a small 22 atom (alanine dipeptide) model and in excess of an order of magnitude for sampling an 882 atom (bovine pancreatic trypsin inhibitor) model, with good scaling with system size subject to the number of modes propagated. NML has been implemented in the open source software PROTOMOL. PMID- 18412480 TI - Protein dynamics from a NMR perspective: networks of coupled rotators and fractional Brownian dynamics. AB - Nuclear magnetic resonance (NMR) has proven to be the most valuable tool for investigating internal dynamics of proteins. In this perspective, the interpretation of NMR relaxation data eventually relies on a model of the motions. In this article, we propose to compare two radically different approaches that aim at describing internal dynamics in proteins. It is shown that the correlation functions predicted by a network of coupled rotators can be interpreted in terms of a heuristic approach based on fractional Brownian dynamics for each of the vectors in the network. Our results are interpreted in terms of the probability distributions of relaxation modes in both processes, the median of which turns out to be the relevant quantity for the comparison of both models. PMID- 18412481 TI - Approximating nonequilibrium processes using a collection of surrogate diffusion models. AB - The surrogate process approximation (SPA) is applied to model the nonequilibrium dynamics of a reaction coordinate (RC) associated with the unfolding and refolding processes of a deca-alanine peptide at 300 K. The RC dynamics, which correspond to the evolution of the end-to-end distance of the polypeptide, are produced by steered molecular dynamics (SMD) simulations and approximated using overdamped diffusion models. We show that the collection of (estimated) SPA models contain structural information "orthogonal" to the RC monitored in this study. Functional data analysis ideas are used to correlate functions associated with the fitted SPA models with the work done on the system in SMD simulations. It is demonstrated that the shape of the nonequilibrium work distributions for the unfolding and refolding processes of deca-alanine can be predicted with functional data analysis ideas using a relatively small number of simulated SMD paths for calibrating the SPA diffusion models. PMID- 18412482 TI - Efficient chain moves for Monte Carlo simulations of a wormlike DNA model: excluded volume, supercoils, site juxtapositions, knots, and comparisons with random-flight and lattice models. AB - We develop two classes of Monte Carlo moves for efficient sampling of wormlike DNA chains that can have significant degrees of supercoiling, a conformational feature that is key to many aspects of biological function including replication, transcription, and recombination. One class of moves entails reversing the coordinates of a segment of the chain along one, two, or three axes of an appropriately chosen local frame of reference. These transformations may be viewed as a generalization, to the continuum, of the Madras-Orlitsky-Shepp algorithm for cubic lattices. Another class of moves, termed T+/-2, allows for interconversions between chains with different lengths by adding or subtracting two beads (monomer units) to or from the chain. Length-changing moves are generally useful for conformational sampling with a given site juxtaposition, as has been shown in previous lattice studies. Here, the continuum T+/-2 moves are designed to enhance their acceptance rate in supercoiled conformations. We apply these moves to a wormlike model in which excluded volume is accounted for by a bond-bond repulsion term. The computed autocorrelation functions for the relaxation of bond length, bond angle, writhe, and branch number indicate that the new moves lead to significantly more efficient sampling than conventional bead displacements and crankshaft rotations. A close correspondence is found in the equilibrium ensemble between the map of writhe computed for pair of chain segments and the map of site juxtapositions or self-contacts. To evaluate the more coarse-grained freely jointed chain (random-flight) and cubic lattice models that are commonly used in DNA investigations, twisting (torsional) potentials are introduced into these models. Conformational properties for a given superhelical density sigma may then be sampled by computing the writhe and using White's formula to relate the degree of twisting to writhe and sigma. Extensive comparisons of contact patterns and knot probabilities of the more coarse-grained models with the wormlike model show that the behaviors of the random-flight model are similar to that of DNA molecules in a solution environment with high ionic strengths, whereas the behaviors of the cubic lattice model with excluded volume are akin to that of DNA molecules under low ionic strengths. PMID- 18412483 TI - Comment on "Markovian approximation in a coarse-grained description of atomic systems" [J. Chem. Phys. 125, 204101 (2006)]. PMID- 18412488 TI - Paraspinal muscle vibration alters dynamic motion of the trunk. AB - Loss in dynamic stability of the low back has been identified as a potential factor in the etiology of low back injuries. A number of factors are important in the ability of a person to maintain an upright trunk posture including the preparatory stiffness of the trunk and the magnitude and timing of the neuromotor response. A neuromotor response requires appropriate sensing of joint motion. In this research, the role of this sensory ability in dynamic performance of the trunk was examined using a simple pendulum model of the trunk with neuromotor feedback. An increased sensory threshold was found to lead to increased torso flexion and increased delay in neuromotor response. This was confirmed experimentally using paraspinal muscle vibration which is known to alter proprioception of the muscle spindle organs. Before, during and after exposure to bilateral, paraspinal muscle vibration for 20 minutes, the dynamic response of subjects to an unexpected torso flexion load was examined. Subjects were found to have a 19.5% slower time to peak muscle activity and a 16.1% greater torso flexion during exposure to paraspinal muscle vibration. Torso flexion remained significantly increased after vibration exposure relative to before exposure. These results suggest that the neuromotor response plays an important role in trunk dynamics. Loss in sensitivity of the sensory system can have a detrimental effect on trunk dynamics, increasing delays in neuromotor response and increasing the motion of the trunk in response to an unexpected load. PMID- 18412489 TI - Study of micromotion in modular acetabular components during gait and subluxation: a finite element investigation. AB - Polyethylene wear after total hip arthroplasty may occur as a result of normal gait and as a result of subluxation and relocation with impact. Relocation of a subluxed hip may impart a moment to the cup creating sliding as well as compression at the cup liner interface. The purpose of the current study is to quantify, by a validated finite element model, the forces generated in a hip arthroplasty as a result of subluxation relocation and compare them to the forces generated during normal gait. The micromotion between the liner and acetabular shell was quantified by computing the sliding track and the deformation at several points of the interface. A finite element analysis of polyethylene liner stress and liner/cup micromotion in total hip arthroplasty was performed under two dynamic profiles. The first profile was a gait loading profile simulating the force vectors developed in the hip arthroplasty during normal gait. The second profile is generated during subluxation and subsequent relocation of the femoral head. The forces generated by subluxation relocation of a total hip arthroplasty can exceed those forces generated during normal gait. The induced micromotion at the cup polyethylene interface as a result of subluxation can exceed micromotion as a result of the normal gait cycle. This may play a significant role in the generation of backsided wear. Minimizing joint subluxation by restoring balance to the hip joint after arthroplasty should be explored as a strategy to minimize backsided wear. PMID- 18412490 TI - Importance of collagen orientation and depth-dependent fixed charge densities of cartilage on mechanical behavior of chondrocytes. AB - The collagen network and proteoglycan matrix of articular cartilage are thought to play an important role in controlling the stresses and strains in and around chondrocytes, in regulating the biosynthesis of the solid matrix, and consequently in maintaining the health of diarthrodial joints. Understanding the detailed effects of the mechanical environment of chondrocytes on cell behavior is therefore essential for the study of the development, adaptation, and degeneration of articular cartilage. Recent progress in macroscopic models has improved our understanding of depth-dependent properties of cartilage. However, none of the previous works considered the effect of realistic collagen orientation or depth-dependent negative charges in microscopic models of chondrocyte mechanics. The aim of this study was to investigate the effects of the collagen network and fixed charge densities of cartilage on the mechanical environment of the chondrocytes in a depth-dependent manner. We developed an anisotropic, inhomogeneous, microstructural fibril-reinforced finite element model of articular cartilage for application in unconfined compression. The model consisted of the extracellular matrix and chondrocytes located in the superficial, middle, and deep zones. Chondrocytes were surrounded by a pericellular matrix and were assumed spherical prior to tissue swelling and load application. Material properties of the chondrocytes, pericellular matrix, and extracellular matrix were obtained from the literature. The loading protocol included a free swelling step followed by a stress-relaxation step. Results from traditional isotropic and transversely isotropic biphasic models were used for comparison with predictions from the current model. In the superficial zone, cell shapes changed from rounded to elliptic after free swelling. The stresses and strains as well as fluid flow in cells were greatly affected by the modulus of the collagen network. The fixed charge density of the chondrocytes, pericellular matrix, and extracellular matrix primarily affected the aspect ratios (height/width) and the solid matrix stresses of cells. The mechanical responses of the cells were strongly location and time dependent. The current model highlights that the collagen orientation and the depth-dependent negative fixed charge densities of articular cartilage have a great effect in modulating the mechanical environment in the vicinity of chondrocytes, and it provides an important improvement over earlier models in describing the possible pathways from loading of articular cartilage to the mechanical and biological responses of chondrocytes. PMID- 18412491 TI - Numerical simulation of pre- and postsurgical flow in a giant basilar aneurysm. AB - Computational modeling of the flow in cerebral aneurysms is an evolving technique that may play an important role in surgical planning. In this study, we simulated the flow in a giant basilar aneurysm before and after surgical takedown of one vertebral artery. Patient-specific geometry and flowrates obtained from magnetic resonance (MR) angiography and velocimetry were used to simulate the flow prior to and after the surgery. Numerical solutions for steady and pulsatile flows were obtained. Highly three-dimensional flows, with strong secondary flows, were computed in the aneurysm in the presurgical and postsurgical conditions. The computational results predicted that occlusion of a vertebral artery would result in a significant increase of the slow flow region formed in the bulge of the aneurysm, where increased particle residence time and velocities lower than 2.5 cms were computed. The region of slow flow was found to have filled with thrombus following surgery. Predictions of numerical simulation methods are consistent with the observed outcome following surgical treatment of an aneurysm. The study demonstrates that computational models may provide hypotheses to test in future studies, and might offer guidance for the interventional treatment of cerebral aneurysms. PMID- 18412492 TI - Modeling the thermal responses of the skin surface during hand-object interactions. AB - The objective of this research is to analyze and model the decreases in skin temperature when the hand makes contact with an object at room temperature so that thermal feedback can be incorporated into haptic displays. A thermal model is proposed that predicts the thermal responses of the skin and object surface as well as the heat flux exchanged during hand-object interactions. The model was evaluated by comparing the theoretical predictions of temperature changes to those experimentally measured using an infrared thermal measurement system. The thermal measurement system was designed to overcome the limitations imposed by contact thermal sensors, and was able to measure skin temperature during contact, together with the contact area and contact force. The experimental results indicated that over the pressure range of 0.73-10.98 kPa, changes in skin temperature were well localized to the contact area and were affected by contact pressure. The pressure in turn influenced both thermal contact resistance and blood flow. Over the range of contact forces typically used in manual exploration, blood perfusion and metabolic heat generation do not appear to have a significant effect on the skin's thermal responses. The theoretical predictions and the measured data were consistent in characterizing the time course and amplitude of the skin temperature change during contact with differences typically being less than 1 degrees C between the two for pressures greater than 4 kPa. These findings indicate that the proposed thermal model is able to characterize and predict the skin temperature responses during hand-object interactions and could be used in a thermal display that simulates the properties of different materials. PMID- 18412493 TI - Continuum mechanics analysis of fracture progression in the vitrified cryoprotective agent DP6. AB - As part of an ongoing effort to study the continuum mechanics effects associated with cryopreservation, the current report focuses on the prediction of fracture formation in cryoprotective agents. Fractures had been previously observed in 1 ml samples of the cryoprotective agent cocktail DP6, contained in a standard 15 ml glass vial, and subjected to various cooling rates. These experimental observations were obtained by means of a cryomacroscope, which has been recently presented by the current research team. High and low cooling rates were found to produce very distinct patterns of cracking. The current study seeks to explain the observed patterns on the basis of stresses predicted from finite element analysis, which relies on a simple viscoelastic constitutive model and on estimates of the critical stress for cracking. The current study demonstrates that the stress, which results in instantaneous fracture at low cooling rates, is consistent with the stress to initiate fracture at high cooling rate. This consistency supports the credibility of the proposed constitutive model and analysis, and the unified criterion for fracturing, that is, a critical stress threshold. PMID- 18412494 TI - Dissection properties of the human aortic media: an experimental study. AB - Aortic dissection occurs frequently and is clinically challenging; the underlying mechanics remain unclear. The present study investigates the dissection properties of the media of 15 human abdominal aortas (AAs) by means of direct tension tests (n=8) and peeling tests (n=12). The direct tension test demonstrates the strength of the media in the radial direction, while the peeling test allows a steady-state investigation of the dissection propagation. To explore the development of irreversible microscopic changes during medial dissection, histological images (n=8) from four AAs at different peeling stages are prepared and analyzed. Direct tension tests of coin-shaped medial specimens result in a radial failure stress of 140.1+/-15.9 kPa (mean+/-SD, n=8). Peeling tests of rectangular-shaped medial strips along the circumferential and axial directions provide peeling force/width ratios of 22.9+/-2.9 mN/mm (n=5) and 34.8+/-15.5 mN/mm (n=7); the related dissection energies per reference area are 5.1+/-0.6 mJ/cm(2) and 7.6+/-2.7 mJ/cm(2), respectively. Although student's t tests indicate that force/width values of both experimental tests are not significantly different (alpha=0.05, p=0.125), the strikingly higher resisting force/width obtained for the axial peeling tests is perhaps indicative of anisotropic dissection properties of the human aortic media. Peeling in the axial direction of the aorta generates a remarkably "rougher" dissection surface with respect to the surface generated by peeling in the circumferential direction. Histological analysis of the stressed specimens reveals that tissue damage spreads over approximately six to seven elastic laminae, which is about 15-18% of the thickness of the abdominal aortic media, which forms a pronounced cohesive zone at the dissection front. PMID- 18412495 TI - Primary stability of cementless stem in THA improved with reduced interfacial gaps. AB - Large interfacial gaps between the stem and the bone in cementless total hip arthroplasty may prevent successful bone ingrowth at the sites, and can also be a passage for wear particles. Furthermore, interfacial gaps between the stem and the bone are believed to compromise the primary stability of the implant. Thus, a broaching method that serves to reduce gaps is expected to give clinically preferable results. A modified broach system with a canal guide is introduced to enhance the accuracy of femoral canal shaping in comparison with the conventional broach system for a Versys fibermetal taper stem. The primary stability of the hip systems and the ratios of the stem surface in contact with the femur were measured in a composite femur model. With the conventional method, an average of 67% of the stem surface was shown to be in contact with the bone, and an average stem micromotion/migration of 35 microm 290 microm was observed under 1000 cycles of stair climbing loads. With the modified method, the stem-bone contact ratio significantly increased to 82% (p<0.05), and the average micromotion/migration reduced to 29 microm 49 microm, respectively (p<0.05 for migration). Our finite element models of the hip systems supported that the difference in micromotion could be attributed to the difference in interfacial contact. Interfacial gaps occurring with the conventional broach system were effectively reduced by the proposed method, resulting in improved primary stability. PMID- 18412496 TI - Design and calibration of load sensing orthopaedic implants. AB - Contact forces and moments act on orthopaedic implants such as joint replacements. The three forces and three moment components can be measured by six internal strain gauges and wireless telemetric data transmission. The accuracy of instrumented implants is restricted by their small size, varying modes of load transfer, and the accuracy of calibration. Aims of this study were to test with finite element studies design features to improve the accuracy, to develop simple but accurate calibration arrangements, and to select the best mathematical method for calculating the calibration constants. Several instrumented implants, and commercial and test transducers were calibrated using different loading setups and mathematical methods. It was found that the arrangement of flexible elements such as bellows or notches between the areas of load transfer and the central sensor locations is most effective to improve the accuracy. Increasing the rigidity of the implant areas, which are fixed in bones or articulate against joint surfaces, is less effective. Simple but accurate calibration of the six force and moment components can be achieved by applying eccentric forces instead of central forces and pure moments. Three different methods for calculating the measuring constants proved to be equally well suited. Employing these improvements makes it possible to keep the average measuring errors of many instrumented implants below 1-2% of the calibration ranges, including cross talk. Additional errors caused by noise of the transmitted signals can be reduced by filtering if this is permitted by the sampling rate and the required frequency content of the loads. PMID- 18412497 TI - Effect of blood flow on near-the-wall mass transport of drugs and other bioactive agents: a simple formula to estimate boundary layer concentrations. AB - Transport of bioactive agents through the blood is essential for cardiovascular regulatory processes and drug delivery. Bioactive agents and other solutes infused into the blood through the wall of a blood vessel or released into the blood from an area in the vessel wall spread downstream of the infusion/release region and form a thin boundary layer in which solute concentration is higher than in the rest of the blood. Bioactive agents distributed along the vessel wall affect endothelial cells and regulate biological processes, such as thrombus formation, atherogenesis, and vascular remodeling. To calculate the concentration of solutes in the boundary layer, researchers have generally used numerical simulations. However, to investigate the effect of blood flow, infusion rate, and vessel geometry on the concentration of different solutes, many simulations are needed, leading to a time-consuming effort. In this paper, a relatively simple formula to quantify concentrations in a tube downstream of an infusion/release region is presented. Given known blood-flow rates, tube radius, solute diffusivity, and the length of the infusion region, this formula can be used to quickly estimate solute concentrations when infusion rates are known or to estimate infusion rates when solute concentrations at a point downstream of the infusion region are known. The developed formula is based on boundary layer theory and physical principles. The formula is an approximate solution of the advection-diffusion equations in the boundary layer region when solute concentration is small (dilute solution), infusion rate is modeled as a mass flux, and there is no transport of solute through the wall or chemical reactions downstream of the infusion region. Wall concentrations calculated using the formula developed in this paper were compared to the results from finite element models. Agreement between the results was within 10%. The developed formula could be used in experimental procedures to evaluate drug efficacy, in the design of drug-eluting stents, and to calculate rates of release of bioactive substances at active surfaces using downstream concentration measurements. In addition to being simple and fast to use, the formula gives accurate quantifications of concentrations and infusion rates under steady-state and oscillatory flow conditions, and therefore can be used to estimate boundary layer concentrations under physiological conditions. PMID- 18412498 TI - Ageing changes in the tensile properties of tendons: influence of collagen fibril volume fraction. AB - Connective tissues are biological composites comprising of collagen fibrils embedded in (and reinforcing) the hydrated proteoglycan-rich (PG) gel within the extracellular matrices (ECMs). Age-related changes to the mechanical properties of tissues are often associated with changes to the structure of the ECM, namely, fibril diameter. However, quantitative attempts to correlate fibril diameter to mechanical properties have yielded inconclusive evidence. Here, we described a novel approach that was based on the rule of mixtures for fiber composites to evaluate the dependence of age-related changes in tendon tensile strength (sigma) and stiffness (E) on the collagen fibril cross-sectional area fraction (rho), which is related to the fibril volume fraction. Tail tendons from C57BL6 mice from age groups 1.6-35.3 months old were stretched to failure to determine sigma and E. Parallel measurements of rho as a function of age were made using transmission electron microscopy. Mathematical models (rule of mixtures) of fibrils reinforcing a PG gel in tendons were used to investigate the influence of rho on ageing changes in sigma and E. The magnitudes of sigma, E, and rho increased rapidly from 1.6 months to 4.0 months (P-values <0.05) before reaching a constant (age independent) from 4.0 months to 29.0 months (P-values >0.05); this trend continued for E and rho (P-values >0.05) from 29.0 months to 35.3 months, but not for sigma, which decreased gradually (P-values <0.05). Linear regression analysis revealed that age-related changes in sigma and E correlated positively to rho (P-values <0.05). Collagen fibril cross-sectional area fraction rho is a significant predictor of ageing changes in sigma and E in the tail tendons of C57BL6 mice. PMID- 18412499 TI - The Phan-Thien and Tanner model applied to thin film spherical coordinates: applications for lubrication of hip joint replacement. AB - The Phan-Thien and Tanner (PTT) model is one of the most widely used rheological models. It can properly describe the common characteristics of viscoelastic non Newtonian fluids. There is evidence that synovial fluid in human joints, which also lubricates artificial joints, is viscoelastic. Modeling the geometry of the total hip replacement, the PTT model is applied in spherical coordinates for a thin confined fluid film. A modified Reynolds equation is developed for this geometry. Several simplified illustrative problems are solved. The effect of the edge boundary condition on load-carrying normal stress is discussed. Solutions are also obtained for a simple squeezing flow. The effect of both the relaxation time and the PTT shear parameter is to reduce the load relative to a Newtonian fluid with the same viscosity. This implies that the Newtonian model is not conservative and may overpredict the load capacity. The PTT theory is a good candidate model to use for joint replacement lubrication. It is well regarded and derivable from molecular considerations. The most important non-Newtonian characteristics can be described with only three primary material parameters. PMID- 18412500 TI - Measurement of the dynamic shear modulus of mouse brain tissue in vivo by magnetic resonance elastography. AB - In this study, the magnetic resonance (MR) elastography technique was used to estimate the dynamic shear modulus of mouse brain tissue in vivo. The technique allows visualization and measurement of mechanical shear waves excited by lateral vibration of the skull. Quantitative measurements of displacement in three dimensions during vibration at 1200 Hz were obtained by applying oscillatory magnetic field gradients at the same frequency during a MR imaging sequence. Contrast in the resulting phase images of the mouse brain is proportional to displacement. To obtain estimates of shear modulus, measured displacement fields were fitted to the shear wave equation. Validation of the procedure was performed on gel characterized by independent rheometry tests and on data from finite element simulations. Brain tissue is, in reality, viscoelastic and nonlinear. The current estimates of dynamic shear modulus are strictly relevant only to small oscillations at a specific frequency, but these estimates may be obtained at high frequencies (and thus high deformation rates), noninvasively throughout the brain. These data complement measurements of nonlinear viscoelastic properties obtained by others at slower rates, either ex vivo or invasively. PMID- 18412501 TI - The effects of femoral fixed body coordinate system definition on knee kinematic description. AB - Understanding the differences in knee kinematic descriptions is important for comparing data from different laboratories and observing small but important changes within a set of knees. The purpose of this study was to identify how differences in fixed body femoral coordinate systems affect the described tibiofemoral and patellofemoral kinematics for cadaveric knee studies with no hip present. Different methods for describing kinematics were evaluated on a set of seven cadaveric knees during walking in a dynamic knee simulator. Three anatomical landmark coordinate systems, a partial helical axis, and an experimental setup-based system were examined. The results showed that flexion extension was insensitive to differences in the kinematic systems tested, internal-external rotation was similar for most femoral coordinate systems although there were changes in absolute position, varus-valgus was the most sensitive to variations in flexion axis direction, and anterior-posterior motion was most sensitive to femoral origin location. Femoral coordinate systems that define the sagittal plane using anatomical landmarks and locate the flexion axis perpendicular to the femur's mechanical axis in the frontal plane were typically similar and described kinematics most consistently. PMID- 18412502 TI - PIV-measured versus CFD-predicted flow dynamics in anatomically realistic cerebral aneurysm models. AB - Computational fluid dynamics (CFD) modeling of nominally patient-specific cerebral aneurysms is increasingly being used as a research tool to further understand the development, prognosis, and treatment of brain aneurysms. We have previously developed virtual angiography to indirectly validate CFD-predicted gross flow dynamics against the routinely acquired digital subtraction angiograms. Toward a more direct validation, here we compare detailed, CFD predicted velocity fields against those measured using particle imaging velocimetry (PIV). Two anatomically realistic flow-through phantoms, one a giant internal carotid artery (ICA) aneurysm and the other a basilar artery (BA) tip aneurysm, were constructed of a clear silicone elastomer. The phantoms were placed within a computer-controlled flow loop, programed with representative flow rate waveforms. PIV images were collected on several anterior-posterior (AP) and lateral (LAT) planes. CFD simulations were then carried out using a well validated, in-house solver, based on micro-CT reconstructions of the geometries of the flow-through phantoms and inlet/outlet boundary conditions derived from flow rates measured during the PIV experiments. PIV and CFD results from the central AP plane of the ICA aneurysm showed a large stable vortex throughout the cardiac cycle. Complex vortex dynamics, captured by PIV and CFD, persisted throughout the cardiac cycle on the central LAT plane. Velocity vector fields showed good overall agreement. For the BA, aneurysm agreement was more compelling, with both PIV and CFD similarly resolving the dynamics of counter rotating vortices on both AP and LAT planes. Despite the imposition of periodic flow boundary conditions for the CFD simulations, cycle-to-cycle fluctuations were evident in the BA aneurysm simulations, which agreed well, in terms of both amplitudes and spatial distributions, with cycle-to-cycle fluctuations measured by PIV in the same geometry. The overall good agreement between PIV and CFD suggests that CFD can reliably predict the details of the intra-aneurysmal flow dynamics observed in anatomically realistic in vitro models. Nevertheless, given the various modeling assumptions, this does not prove that they are mimicking the actual in vivo hemodynamics, and so validations against in vivo data are encouraged whenever possible. PMID- 18412503 TI - Carousel effect in alveolar models. AB - Experimental work over the past decade has shown that recirculation in alveoli substantially increases the transport of particles. We have previously shown that, for nondiffusing passive particles, this can be understood with the aid of Moffatt's famous corner flow model. Without wall motion, passive particles recirculate in a regular fashion and no chaos exists; however, wall motion produces extensive chaotic flow. Aerosols typically do not follow this flow as they are fundamentally different from fluid particles. Here, we construct a simple model to study diffusing particles in the presence of recirculation. We assume that all particles are passive, that is to say that they do not significantly alter the underlying flow. In particular, we consider particles with high Peclet number and neglect inertial effects. We modify the Lagrangian system for corner eddies to accommodate diffusing particles. Particle transport is governed by Langevin equations. Ensembles of diffusing particles are tracked by numerical integration. We show that transport of diffusing particles is enhanced by sufficiently strong underlying recirculation through a mechanism that we call the "carousel effect." However, as the corner is approached, the recirculation rapidly decreases in intensity, favoring motion by diffusion. Far from the corner's apex, recirculation dominates. For real alveoli, the model indicates that sufficiently strong recirculation can enhance transport of diffusing particles through the carousel effect. PMID- 18412504 TI - Wavelet analysis of head acceleration response under dirac excitation for early oedema detection. AB - The present work deals with the application of an innovative in-house developed wavelet-based methodology for the analysis of the acceleration responses of a human head complex model as a simulated diffused oedema progresses. The human head complex has been modeled as a structure consisting of three confocal prolate spheroids, whereas the three defined regions by the system of spheroids, from the outside to the inside, represent the scull, the region of cerebrospinal fluid, and the brain tissue. A Dirac-like pulse has been used to excite the human head complex model and the acceleration response of the system has been calculated and analyzed via the wavelet-based methodology. For the purpose of the present analysis, a wave propagation commercial finite element code, LS-DYNA 3D, has been used. The progressive diffused oedema was modeled via consecutive increases in brain volume accompanied by a decrease in brain density. It was shown that even a small increase in brain volume (at the level of 0.5%) can be identified by the effect it has on the vibration characteristics of the human head complex. More precisely, it was found that for some of the wavelet decomposition levels, the energy content changes monotonically as the brain volume increases, thus providing a useful index of monitoring an oncoming brain oedema before any brain damage appears due to uncontrolled intracranial hypertension. For the purpose of the present work and for the levels of brain volume increase considered in the present analysis, no pressure increase was assumed into the cranial vault and, associatively, no brain compliance variation. PMID- 18412505 TI - Numerical study of the uniformity of balloon-expandable stent deployment. AB - Stents are small tubelike structures, implanted in coronary and peripheral arteries to reopen narrowed vessel sections. This endovascular intervention remains suboptimal, as the success rate is limited by restenosis. This renarrowing of a stented vessel is related to the arterial injury caused by stent artery and balloon-artery interactions, and a local subsequent inflammatory process. Therefore, efforts to optimize the stent deployment remain very meaningful. Several authors have studied with finite element modeling the mechanical behavior of balloon-expandable stents, but none of the proposed models incorporates the folding pattern of the balloon. We developed a numerical model in which the CYPHER stent is combined with a realistic trifolded balloon. In this paper, the impact of several parameters such as balloon length, folding pattern, and relative position of the stent with respect to the balloon catheter on the free stent expansion has been investigated. Quantitative validation of the modeling strategy shows excellent agreement with data provided by the manufacturer and, therefore, the model serves as a solid basis for further investigations. The parametric analyses showed that both the balloon length and the folding pattern have a considerable influence on the uniformity and symmetry of the transient stent expansion. Consequently, this approach can be used to select the most appropriate balloon length and folding pattern for a particular stent design in order to optimize the stent deployment. Furthermore, it was demonstrated that small positioning inaccuracies may change the expansion behavior of a stent. Therefore, the placement of the stent on the balloon catheter should be accurately carried out, again in order to decrease the endothelial damage. PMID- 18412506 TI - In vitro fatigue failure of cemented acetabular replacements: a hip simulator study. AB - Although hip simulators for in vitro wear testing of prosthetic materials used in total hip arthroplasty (THA) have been available for a number of years, similar equipment has yet to appear for endurance testing of fixation in cemented THA, despite considerable evidence of late aseptic loosening as one of the most significant failure mechanisms in this type of replacements. An in vitro study of fatigue behavior in cemented acetabular replacements has been carried out, utilizing a newly developed hip simulator. The machine was designed to simulate the direction and the magnitude of the hip contact force under typical physiological loading conditions, including normal walking and stair climbing, as reported by Bergmann et al. (2001, Hip 98, Freie Universitaet, Berlin). A 3D finite element analysis has been carried out to validate the function of the hip simulator and to evaluate the effects of boundary conditions and geometry of the specimen on the stress distribution in the cement mantle. Bovine pelvic bones were implanted with a Charnley cup, using standard manual cementing techniques. Experiments were carried out under normal walking and descending stairs loading conditions with selected load levels from a body weight of 75-125 kg. Periodically, the samples were removed from the test rigs to allow CT scanning for the purpose of monitoring damage development in the cement fixation. The hip simulator was found to be satisfactory in reproducing the hip contact force during normal walking and stair climbing, as reported by Bergmann et al. Finite element analysis shows that the stress distributions in the cement mantle and at the bone-cement interface are largely unaffected by the geometry and the boundary conditions of the model. Three samples were tested up to 17 x 10(6) cycles and sectioned post-testing for microscopic studies. Debonding at the bone-cement interface of various degrees in the posterior-superior quadrant was revealed in these samples, and the location of the failures corresponds to the highest stressed region from the finite-element analysis. Preliminary experimental results from a newly developed hip simulator seem to suggest that debonding at the bone-cement interface is the main failure mechanism in cemented acetabular replacements, and descending stairs seem to be more detrimental than normal walking or ascending stairs with regard to fatigue integrity of cement fixation. PMID- 18412507 TI - Biomechanical properties of abdominal organs in vivo and postmortem under compression loads. AB - Accurate knowledge of biomechanical characteristics of tissues is essential for developing realistic computer-based surgical simulators incorporating haptic feedback, as well as for the design of surgical robots and tools. As simulation technologies continue to be capable of modeling more complex behavior, an in vivo tissue property database is needed. Most past and current biomechanical research is focused on soft and hard anatomical structures that are subject to physiological loading, testing the organs in situ. Internal organs are different in that respect since they are not subject to extensive loads as part of their regular physiological function. However, during surgery, a different set of loading conditions are imposed on these organs as a result of the interaction with the surgical tools. Following previous research studying the kinematics and dynamics of tool/tissue interaction in real surgical procedures, the focus of the current study was to obtain the structural biomechanical properties (engineering stress-strain and stress relaxation) of seven abdominal organs, including bladder, gallbladder, large and small intestines, liver, spleen, and stomach, using a porcine animal model. The organs were tested in vivo, in situ, and ex corpus (the latter two conditions being postmortem) under cyclical and step strain compressions using a motorized endoscopic grasper and a universal-testing machine. The tissues were tested with the same loading conditions commonly applied by surgeons during minimally invasive surgical procedures. Phenomenological models were developed for the various organs, testing conditions, and experimental devices. A property database-unique to the literature-has been created that contains the average elastic and relaxation model parameters measured for these tissues in vivo and postmortem. The results quantitatively indicate the significant differences between tissue properties measured in vivo and postmortem. A quantitative understanding of how the unconditioned tissue properties and model parameters are influenced by time postmortem and loading condition has been obtained. The results provide the material property foundations for developing science-based haptic surgical simulators, as well as surgical tools for manual and robotic systems. PMID- 18412508 TI - Modeling of dynamic fracture and damage in two-dimensional trabecular bone microstructures using the cohesive finite element method. AB - Trabecular bone fracture is closely related to the trabecular architecture, microdamage accumulation, and bone tissue properties. Micro-finite-element models have been used to investigate the elastic and yield properties of trabecular bone but have only seen limited application in modeling the microstructure dependent fracture of trabecular bone. In this research, dynamic fracture in two dimensional (2D) micrographs of ovine (sheep) trabecular bone is modeled using the cohesive finite element method. For this purpose, the bone tissue is modeled as an orthotropic material with the cohesive parameters calculated from the experimental fracture properties of the human cortical bone. Crack propagation analyses are carried out in two different 2D orthogonal sections cut from a three dimensional 8 mm diameter cylindrical trabecular bone sample. The two sections differ in microstructural features such as area fraction (ratio of the 2D space occupied by bone tissue to the total 2D space), mean trabecula thickness, and connectivity. Analyses focus on understanding the effect of the rate of loading as well as on how the rate variation interacts with the microstructural features to cause anisotropy in microdamage accumulation and in the fracture resistance. Results are analyzed in terms of the dependence of fracture energy dissipation on the microstructural features as well as in terms of the changes in damage and stresses associated with the bone architecture variation. Besides the obvious dependence of the fracture behavior on the rate of loading, it is found that the microstructure strongly influences the fracture properties. The orthogonal section with lesser area fraction, low connectivity, and higher mean trabecula thickness is more resistant to fracture than the section with high area fraction, high connectivity, and lower mean trabecula thickness. In addition, it is found that the trabecular architecture leads to inhomogeneous distribution of damage, irrespective of the symmetry in the applied loading with the fracture of the entire bone section rapidly progressing to bone fragmentation once the accumulated damage in any trabeculae reaches a critical limit. PMID- 18412509 TI - Development and validation of subject-specific finite element models for blunt trauma study. AB - This study developed and validated finite element (FE) models of swine and human thoraxes and abdomens that had subject-specific anatomies and could accurately and efficiently predict body responses to blunt impacts. Anatomies of the rib cage, torso walls, thoracic, and abdominal organs were reconstructed from X-ray computed tomography (CT) images and extracted into geometries to build FE meshes. The rib cage was modeled as an inhomogeneous beam structure with geometry and bone material parameters determined directly from CT images. Meshes of soft components were generated by mapping structured mesh templates representative of organ topologies onto the geometries. The swine models were developed from and validated by 30 animal tests in which blunt insults were applied to swine subjects and CT images, chest wall motions, lung pressures, and pathological data were acquired. A comparison of the FE calculations of animal responses and experimental measurements showed a good agreement. The errors in calculated response time traces were within 10% for most tests. Calculated peak responses showed strong correlations with the experimental values. The stress concentration inside the ribs, lungs, and livers produced by FE simulations also compared favorably to the injury locations. A human FE model was developed from CT images from the Visible Human project and was scaled to simulate historical frontal and side post mortem human subject (PMHS) impact tests. The calculated chest deformation also showed a good agreement with the measurements. The models developed in this study can be of great value for studying blunt thoracic and abdominal trauma and for designing injury prevention techniques, equipments, and devices. PMID- 18412510 TI - Mechanical stresses in abdominal aortic aneurysms: influence of diameter, asymmetry, and material anisotropy. AB - Biomechanical studies suggest that one determinant of abdominal aortic aneurysm (AAA) rupture is related to the stress in the wall. In this regard, a reliable and accurate stress analysis of an in vivo AAA requires a suitable 3D constitutive model. To date, stress analysis conducted on AAA is mainly driven by isotropic tissue models. However, recent biaxial tensile tests performed on AAA tissue samples demonstrate the anisotropic nature of this tissue. The purpose of this work is to study the influence of geometry and material anisotropy on the magnitude and distribution of the peak wall stress in AAAs. Three-dimensional computer models of symmetric and asymmetric AAAs were generated in which the maximum diameter and length of the aneurysm were individually controlled. A five parameter exponential type structural strain-energy function was used to model the anisotropic behavior of the AAA tissue. The anisotropy is determined by the orientation of the collagen fibers (one parameter of the model). The results suggest that shorter aneurysms are more critical when asymmetries are present. They show a strong influence of the material anisotropy on the magnitude and distribution of the peak stress. Results confirm that the relative aneurysm length and the degree of aneurysmal asymmetry should be considered in a rupture risk decision criterion for AAAs. PMID- 18412511 TI - Contributions of the individual muscles of the shoulder to glenohumeral joint stability during abduction. AB - The aim of this study was to determine the relative contributions of the deltoid and rotator cuff muscles to glenohumeral joint stability during arm abduction. A three-dimensional model of the upper limb was used to calculate the muscle and joint-contact forces at the shoulder for abduction in the scapular plane. The joints of the shoulder girdle-sternoclavicular joint, acromioclavicular joint, and glenohumeral joint-were each represented as an ideal three degree-of-freedom ball-and-socket joint. The articulation between the scapula and thorax was modeled using two kinematic constraints. Eighteen muscle bundles were used to represent the lines of action of 11 muscle groups spanning the glenohumeral joint. The three-dimensional positions of the clavicle, scapula, and humerus during abduction were measured using intracortical bone pins implanted into one subject. The measured bone positions were inputted into the model, and an optimization problem was solved to calculate the forces developed by the shoulder muscles for abduction in the scapular plane. The model calculations showed that the rotator cuff muscles (specifically, supraspinatus, subscapularis, and infraspinatus) by virtue of their lines of action are perfectly positioned to apply compressive load across the glenohumeral joint, and that these muscles contribute most significantly to shoulder joint stability during abduction. The middle deltoid provides most of the compressive force acting between the humeral head and the glenoid, but this muscle also creates most of the shear, and so its contribution to joint stability is less than that of any of the rotator cuff muscles. PMID- 18412512 TI - Heat transfer to blood flow in a small tube. AB - Blood flow in a small tube (30-1000 mum) can be successfully modeled by the two fluid model. The fully developed, constant heat flux convective heat transfer problem is studied. The velocity and temperature profiles are determined in closed form. Formulas for friction-factor-Reynolds number product, axial temperature gradient, and Nusselt number are found. PMID- 18412513 TI - Development of a testing methodology to quantify bone load transfer patterns for multiple stemmed implants in a single bone with an application in the distal ulna. AB - Optimal parameters for many orthopaedic implants, such as stem length and material, are unknown. Geometry and mechanical properties of bone can vary greatly amongst cadaveric specimens, requiring a large number of specimens to test design variations. This study aimed to develop an experimental methodology to measure bone strains as a function of multiple implant stem designs in a single specimen, and evaluate its efficacy in the distal ulna. Eight fresh-frozen cadaveric ulnae were each instrumented with 12 uniaxial strain gauges on the medial and lateral surfaces of the bone. The proximal portion of each ulna was cemented in a custom-designed jig that allowed a medially directed force to be applied to the distal articular surface. An implant with a finely threaded stem was cemented into the canal by an experienced upper extremity orthopaedic surgeon. Six loads (5-30 N) were applied sequentially to the lateral surface of the prosthetic head using a materials testing machine. Testing was repeated after breaking the stem-cement bond, and after removing and reinserting the stem several times into the threaded cement mantle. Near the end of the testing period, the initial stem was reinserted and data were collected to determine if there was any change in bone properties or testing setup over time. Finally, a smooth stem was inserted for comparison to the threaded stem. Strain varied linearly with load (R(2)> or =0.99) for all testing scenarios. Bending strains were not affected by breaking the stem-cement bond (P=0.7), testing durations up to 18 h (P=0.7), nor the presence of threads when compared to a smooth stem (P>0.4). Furthermore, for all gauges, there was no interaction between the effect of the threads and level of applied load (P>0.1). This methodology should prove to be useful to compare stem designs of varying lengths and materials in the same bone, allowing for a direct comparison between implant designs for the ulna and other bones subjected primarily to bending loads. Furthermore, it will minimize the need for large numbers of specimens to test multiple implant designs. The ultimate goal of using this protocol is to optimize implant stem properties, such as length and material, with respect to load transfer. PMID- 18412515 TI - The effects of pertussis toxin-treatment on integrated vasoactive response of vascular system in spontaneously hypertensive rats. AB - We investigated the effect of pertussis toxin (PTX) on hypotensive response induced by acetylcholine (ACh) and bradykinin (BK) and on noradrenaline (NA) induced pressor response in spontaneously hypertensive rats (SHR). Fifteen-week old Wistar rats and age-matched SHR were used. Half of SHR received PTX (10 microg/kg/i.v.) and the experiments were performed 48 h later. After the anesthesia the right carotid artery was cannulated in order to record blood pressure (BP). The hypotensive response to ACh was enhanced in SHR compared to Wistar rats. After pretreatment of SHR with PTX the hypotensive response to ACh was reduced compared to untreated SHR and it was also diminished in comparison to Wistar rats. Similarly, the hypotensive response to BK was also decreased after PTX pretreatment. The pressor response to NA was increased in SHR compared to Wistar rats. NA-induced pressor response was considerably decreased after PTX pretreatment compared to untreated SHR. In conclusion, the enhancement of hypotensive and pressor responses in SHR was abolished after PTX pretreatment. Our results suggested that the activation of PTX-sensitive inhibitory G(i) proteins is involved in the regulation of integrated vasoactive responses in SHR and PTX pretreatment could be effectively used for modification of BP regulation in this type of experimental hypertension. PMID- 18412516 TI - Characterization of complete particles (VSV-G/SIN-GFP) and empty particles (VSV G/EMPTY) in human immunodeficiency virus type 1-based lentiviral products for gene therapy: potential applications for improvement of product quality and safety. AB - Lentiviral vectors persist in the host and are therefore ideally suited for long term gene therapy. To advance the use of lentiviral vectors in humans, improvement of their production, purification, and characterization has become increasingly important and challenging. In addition to cellular contaminants derived from packaging cells, empty particles without therapeutic function are the major impurities that compromise product safety and efficacy. Removal of empty particles is difficult because of their innate similarity in particle size and protein composition to the complete particles. We propose that comparison of the properties of lentiviral products with those of purposely expressed empty particles may reveal potential differences between empty and complete particles. For this, three forms of recombinant lentiviral samples, that is, recombinant vesicular stomatitis virus glycoprotein (VSV-G) proteins, empty particles (VSV G/Empty), and complete particles (VSV-G/SIN-GFP) carrying viral RNA, were purified by size-exclusion chromatography (SEC). The SEC-purified samples were further analyzed by immunoblotting with six antibodies to examine viral and cellular proteins associated with the particles. This study has demonstrated, for the first time, important differences between VSV-G/Empty particles and complete VSV-G/SIN-GFP particles. Differences include the processing of Gag protein and the inclusion of cellular proteins in the particles. Our findings support the development of improved production, purification, and characterization methods for lentiviral products. PMID- 18412517 TI - Isoforms of bone alkaline phosphatase, stem cells, and osteoblast phenotypes. PMID- 18412520 TI - Thoughts on protecting veterinary responders following natural disasters. PMID- 18412521 TI - What is your diagnosis? Duroc gilt evaluated for a 1-month history of left forelimb lameness. PMID- 18412522 TI - Diagnostic imaging in veterinary dental practice: right mandibular first molar. PMID- 18412523 TI - Use of systemically administered lidocaine in horses with gastrointestinal tract disease. PMID- 18412524 TI - Seroprevalences of feline leukemia virus and feline immunodeficiency virus in cats with abscesses or bite wounds and rate of veterinarian compliance with current guidelines for retrovirus testing. AB - OBJECTIVE: To determine the seroprevalences of and seroconversion rates for FeLV and FIV infection in cats treated for bite wounds and cutaneous abscesses and to evaluate compliance with recommendations to determine the retrovirus infection status of cats at acquisition and 60 days after a high-risk event. DESIGN: Prospective study. ANIMALS: 967 cats from 134 veterinary practices in 30 states. PROCEDURES: Cats with bite wounds or abscesses were evaluated by use of a point of-care immunoassay for blood-borne FeLV antigen and FIV antibody. Veterinarians were asked to retest cats approximately 60 days later to determine whether seronegative cats had seroconverted after injury. RESULTS: The combined FeLV-FIV status of only 96 (9.9%) cats was known prior to wound treatment. At the time of treatment, 187 (19.3%) cats were seropositive for 1 or both viruses. Age (adult), sex (male), history of cutaneous wounds, and outdoor access were significantly associated with seropositivity. At 73 of 134 (54.5%) veterinary practices, retesting of cats for retrovirus infection status was recommended to owners of 478 cats. Only 64 (13.4%) cats were retested; of these, 3 of 58 (5.2%) cats that were initially seronegative for FIV antibody seroconverted. CONCLUSIONS AND CLINICAL RELEVANCE: A high proportion of cats with abscesses or bite wounds were seropositive for FeLV antigen or FIV antibody. Compliance with recommendations to test cats for retrovirus infection status at acquisition or after treatment for injury was low. The FeLV-FIV infection status of cats with potential fight wounds should be determined at time of treatment and again 60 days later. PMID- 18412526 TI - Attitudes toward and perceptions of free-roaming cats among individuals living in Ohio. AB - OBJECTIVE: To characterize attitudes toward and perceptions of free-roaming cats among individuals living in Ohio. DESIGN: Cross-sectional study. SAMPLE POPULATION: Households in Ohio. PROCEDURES: A random-digit-dial telephone survey was performed, and 1,250 households were contacted. RESULTS: 703 of the 1,250 (56.2%) households completed interviews. Five hundred fifty-three (78.7%) participants reported seeing free-roaming cats at least occasionally, and 184 (26.2%) reported having fed free-roaming cats during the previous year. However, only 42 (22.8%) participants who fed free-roaming cats had ever taken one to a veterinarian, and 43 (23.4%) participants who fed free-roaming cats reported that at least one of the free-roaming cats had produced a litter in the preceding year. Differences existed between cat owners and other participants and among urban, suburban, and rural residents in regard to their attitudes toward free roaming cats and the need for government regulations. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that free-roaming cats were common in Ohio, but that attitudes toward and perceptions of free-roaming cats differed between cat owners and other participants and among participants grouped on the basis of residential area. Thus, developing statewide approaches for regulating free-roaming cats may be challenging or unrealistic. PMID- 18412527 TI - Suspected albuterol toxicosis in a dog. AB - CASE DESCRIPTION: A 6-year-old male castrated Shetland Sheepdog was evaluated because of severe hypokalemia and progressive paresis. CLINICAL FINDINGS: Physical examination revealed fever, tachypnea, mydriasis, hyperemic mucous membranes, severe forelimb paresis, and hind limb paraplegia. The dog had superficial and deep pain sensation in all 4 limbs. Forelimb spinal reflexes were considered normal, but hind limb reflexes were normal to slightly hyperreflexive. The panniculus reflex was considered to be normal, and cranial nerve reflexes were intact. A CBC revealed mild leukocytosis and erythrocytosis, and serum biochemical analysis revealed severe hypokalemia. Thoracic and abdominal imaging did not reveal relevant findings. Blood pressure and ECG findings were within reference limits. Questioning of the owner revealed possible exposure to albuterol via ingestion of medication intended for the owner's horse. Results of serum testing via immunoassay were suggestive of albuterol toxicosis. TREATMENT AND OUTCOME: Treatment included IV administration of an electrolyte solution and supplemental potassium chloride. The rate of potassium chloride supplementation was slowly decreased as serum potassium concentration increased. No other medical intervention was required, and the dog made a rapid and complete recovery. CLINICAL RELEVANCE: Ingestion of albuterol can lead to profound physical and serum biochemical abnormalities. Appropriate historical information should be obtained to identify possible sources and routes of exposure to intoxicants. Albuterol-induced hypokalemia can be successfully managed medically. PMID- 18412529 TI - Acquired pulmonary artery stenosis in four dogs. AB - CASE DESCRIPTION: 4 dogs with acquired pulmonary artery stenosis (PAS) were examined for various clinical signs. One was a mixed-breed dog with congenital valvular PAS that subsequently developed peripheral PAS, one was a Golden Retriever with pulmonary valve fibrosarcoma, one was a Pembroke Welsh Corgi in which the left pulmonary artery had inadvertently been ligated during surgery for correction of patent ductus arteriosus, and one was a Boston Terrier with a heart base mass compressing the pulmonary arteries. CLINICAL FINDINGS: All 4 dogs were evaluated with 2-dimensional and Doppler echocardiography to characterize the nature and severity of the stenoses; other diagnostic tests were also performed. TREATMENT AND OUTCOME: The mixed-breed dog with valvular and peripheral PAS was euthanized, surgical resection of the pulmonic valve mass was performed in the Golden Retriever, corrective surgery was performed on the Pembroke Welsh Corgi with left pulmonary artery ligation, and the Boston Terrier with the heart-base mass was managed medically. CLINICAL RELEVANCE: Acquired PAS in dogs may manifest as a clinically silent heart murmur, syncope, or right-sided heart failure. The diagnosis is made on the basis of imaging findings, particularly results of 2 dimensional and Doppler echocardiography. Treatment may include surgical, interventional, or medical modalities and is targeted at resolving the inciting cause. PMID- 18412530 TI - Goitrous hypothyroidism associated with treatment with trimethoprim sulfamethoxazole in a young dog. AB - CASE DESCRIPTION: A 16-week-old female Boxer that had been treated for 5 weeks with trimethoprim-sulfamethoxazole and chloramphenicol because of aspiration pneumonia was evaluated for bilaterally symmetric masses in the subcutaneous tissues of the ventral neck, in the region of the larynx. CLINICAL FINDINGS: Fine needle aspirates were obtained from the neck masses; cytologic examination revealed well-differentiated thyroid epithelial tissue. A blood sample was collected for serum biochemical and thyroid function analyses. Mild hyperphosphatemia, severe hypercholesterolemia, mild hyperkalemia, and a mild increase in creatine kinase activity were identified. Serum concentration of total thyroxine was less than the lower reference limit, and that of thyroid stimulating hormone was greater than the upper reference limit. Findings were consistent with a diagnosis of clinical hypothyroidism in a skeletally immature dog. TREATMENT AND OUTCOME: Treatment with trimethoprim-sulfamethoxazole was discontinued. The dog was reevaluated 3 weeks later, at which time the neck masses were markedly decreased in size. Serum concentrations of cholesterol and potassium were lower; serum concentrations of total thyroxine and thyroid stimulating hormone were near or within respective reference ranges. Age appropriate increases in serum phosphorus concentration and serum alkaline phosphatase activity were also detected. CLINICAL RELEVANCE: To the authors' knowledge, this is the first report of antimicrobial-induced goiter in a dog. Cytologic examination of fine-needle aspirates and interpretation of data from serum biochemical and thyroid function analyses were needed to obtain a definitive diagnosis. Practitioners should include goiter among the differential diagnoses for ventral neck swellings in young dogs receiving potentiated sulfonamide antimicrobials. PMID- 18412532 TI - Results of excision of thymoma in cats and dogs: 20 cases (1984-2005). AB - OBJECTIVE: To provide long-term follow-up information for a series of dogs and cats with invasive and noninvasive thymomas treated by excision alone. DESIGN: Retrospective case series. ANIMALS: 9 cats and 11 dogs with thymoma. PROCEDURES: Medical records were reviewed. The following factors were analyzed for their effect on prognosis: age of dog or cat, invasiveness of the tumor, percentage of lymphocytes in the mass (percentage lymphocyte composition) on histologic evaluation, and mitotic index of the mass. RESULTS: All patients were treated with excision of the tumor alone. Median overall survival time for the cats was 1,825 days, with a 1-year survival rate of 89% and a 3-year survival rate of 74%. Median overall survival time for the dogs was 790 days, with a 1-year survival rate of 64% and a 3-year survival rate of 42%. Recurrence of thymoma was observed in 2 cats and 1 dog, and a second surgery was performed in each, with subsequent survival times of 5, 3, and 4 years following the first surgery. Percentage lymphocyte composition of the mass was the only factor that was significantly correlated with survival time; animals with a high percentage of lymphocytes lived longer. CONCLUSIONS AND CLINICAL RELEVANCE: Results of this study indicated that most cats and dogs with thymomas did well after excision. Even cats and dogs with invasive masses that survived the surgery and the few cats and dogs with recurrent thymomas or paraneoplastic syndromes had a good long-term outcome. Excision should be considered an effective treatment option for dogs and cats with thymomas. PMID- 18412533 TI - Analysis of prognostic factors associated with injection-site sarcomas in cats: 57 cases (2001-2007). AB - OBJECTIVE: To identify prognostic factors in cats with injection-site sarcomas (ISSs). DESIGN: Retrospective case series. ANIMALS: 57 cats with ISSs. PROCEDURES: Medical records of cats were reviewed with regard to sex, age, anatomic site of tumor, tumor size, histologic grade, excision of a primary tumor versus excision of a recurrent ISS, use of excision alone versus excision plus adjuvant therapy, local tumor recurrence, and development of distant metastasis to predict overall survival time (ie, time from tumor excision to death). RESULTS: In univariate analyses, local recurrence and development of distant metastasis were significantly associated with survival time in cats. On multivariate analysis, development of distant metastasis remained a significant prognostic factor. Histologic grade was associated with distant metastasis, with cats having grade 3 tumors being significantly more likely to develop metastasis than cats with grade 1 and 2 tumors. Factors associated with local recurrence of ISSs were not identified. CONCLUSIONS AND CLINICAL RELEVANCE: The development of distant metastasis, which may occur later during the course of the disease, was identified as a prognostic factor for overall survival time in cats with ISSs. In addition, cats with histologic grade 3 ISSs should be considered for further interventional studies with chemotherapy to prevent the high rate of distant metastasis. PMID- 18412534 TI - Lomustine for treatment of mast cell tumors in cats: 38 cases (1999-2005). AB - OBJECTIVE: To determine clinical activity and toxic effects of lomustine when used to treat cats with mast cell tumors (MCTs). DESIGN: Retrospective case series. ANIMALS: 38 cats with measurable, histologically or cytologically confirmed MCTs treated with lomustine at a dosage > or = 50 mg/m(2). PROCEDURES: Medical records were reviewed to determine response to treatment and evidence of drug toxicoses. The Kaplan-Meier method was used to estimate remission duration. RESULTS: 26 cats had cutaneous MCTs, 7 had MCTs of the mesenteric lymph nodes, 2 had gastrointestinal tract MCTs, 2 had hepatic MCTs, and 1 had MCTs involving multiple organs. Targeted lomustine dosage was 50 mg/m(2) in 22 cats and 60 mg/m(2) in 16 cats. Median administered dosage of lomustine was 56 mg/m(2) (range, 48 to 65 mg/m(2)), and median number of doses administered was 2 (range, 1 to 12). Seven cats had a complete response and 12 had a partial response, for an overall response rate of 50%. Median response duration was 168 days (range, 25 to 727 days). The most common toxicoses were neutropenia and thrombocytopenia. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that lomustine had activity against MCTs in cats and was well tolerated. Further, findings suggested that treatment with lomustine should be considered for cats with MCTs for which local treatment is not an option. PMID- 18412535 TI - Hematuria and transitional cell papilloma of the renal pelvis treated via unilateral nephrectomy in an alpaca. AB - CASE DESCRIPTION: An 11-year-old 72-kg (158-lb) sexually intact female alpaca was examined for diagnosis and treatment of hematuria of 4 months' duration. CLINICAL FINDINGS: Pigmenturia was detected by the owner when the alpaca was 8 months pregnant. Radiographic, ultrasonographic, vaginal speculum, and cystoscopic evaluation of the urinary tract revealed normal vaginal and urethral epithelia and increased bladder vessel tortuosity, with pulses of hemorrhage from the left ureter. Regenerative anemia and mild leukopenia were detected and serum urea nitrogen and creatinine concentrations were within reference ranges. TREATMENT AND OUTCOME: Chronic hematuria resolved after unilateral nephrectomy of the left kidney, and no dysfunction was detected in the remaining kidney. Histologic evaluation of the kidney revealed a transitional cell tumor in the renal pelvis. CLINICAL RELEVANCE: Although anemia is common in South American camelids, hematuria is an uncommon sign of this condition. Chronic urinary tract infection, toxin ingestion, and neoplasia causing hematuria or hemoglobinuria should be considered in South American camelids with pigmenturia. Thorough and systematic evaluation of the urinary tract should be performed to locate the site of hemorrhage to treat hematuria appropriately. PMID- 18412536 TI - Targeted drug-aerosol delivery in the human respiratory system. AB - Inhalation of drug aerosols is a modern pathway to combat lung diseases. It is also becoming the preferred route for insulin delivery, pain management, cancer therapy, and nanotherapeutics. Popular delivery devices include nebulizers, metered-dose inhalers, and dry-powder inhalers. They are all nondirectional and hence have typically low particle deposition efficiencies in desired nasal or lung areas. Thus, for specific disease treatment with costly and/or aggressive medicine, it is necessary to provide targeted drug-aerosol delivery to predetermined sites in the human respiratory system. Experimental measurements and computer models of particle transport and deposition in nasal and lung airway models are presented. Furthermore, the underlying methodology and performance of pressurized metered dose inhalers as well as new smart inhaler systems are discussed. To maximize respiratory drug delivery to specific sites, an optimal combination of particle characteristics, inhalation waveform, particle release position, and drug-aerosol dosage has to be achieved. PMID- 18412537 TI - Single-molecule studies of protein folding. AB - Although protein-folding studies began several decades ago, it is only recently that the tools to analyze protein folding at the single-molecule level have been developed. Advances in single-molecule fluorescence and force spectroscopy techniques allow investigation of the folding and dynamics of single protein molecules, both at equilibrium and as they fold and unfold. The experiments are far from simple, however, both in execution and in interpretation of the results. In this review, we discuss some of the highlights of the work so far and concentrate on cases where comparisons with the classical experiments can be made. We conclude that, although there have been relatively few startling insights from single-molecule studies, the rapid progress that has been made suggests that these experiments have significant potential to advance our understanding of protein folding. In particular, new techniques offer the possibility to explore regions of the energy landscape that are inaccessible to classical ensemble measurements and, perhaps, to observe rare events undetectable by other means. PMID- 18412538 TI - Advances in single-molecule fluorescence methods for molecular biology. AB - Ever since their introduction two decades ago, single-molecule (SM) fluorescence methods have matured and branched out to address numerous biological questions, which were inaccessible via ensemble measurements. Among the current arsenal, SM fluorescence techniques have capabilities of probing the dynamic interactions of nucleic acids and proteins via Forster (fluorescence) resonance energy transfer (FRET), tracking single particles over microns of distances, and deciphering the rotational motion of multisubunit systems. In this exciting era of transitioning from in vitro to in vivo and in situ conditions, it is anticipated that SM fluorescence methodology will become a common tool of molecular biology. PMID- 18412540 TI - Development of affinity columns for the removal of high-abundance proteins in cerebrospinal fluid. AB - Various approaches for removal of high-abundance components in body fluids are currently available. While most methods are constructed for plasma depletion, there is a need for body-fluid-specific strategies. The aim of the present study was to design an affinity matrix suitable for the depletion of high-abundance proteins in CSF (cerebrospinal fluid). Hence, molecules with specific affinity towards proteins present at high concentration in CSF were desired. Affibody molecules are specific binders of small size that have shown high stability under various conditions and are therefore good candidates for such a matrix. The protein composition in CSF resembles that in plasma. However, 20% of the proteins are brain-derived and are therefore present in higher proportions in CSF than in plasma, whereas larger plasma-derived proteins are less abundant in CSF. Therefore five high-abundance CSF proteins were chosen for the design of a CSF specific depletion setup. Affibody molecules with specificity towards HSA (human serum albumin), IgG, transferrin and transthyretin were combined in an affinity column. In addition, polyclonal antibodies against cystatin C were coupled to chromatographic beads and packed in a separate column. Highly reproducible and efficient removal of the five target proteins was observed. The proportion of depleted proteins were estimated to be 99, 95, 74, 92 and 83% for HSA, IgG, transferrin, transthyretin and cystatin C respectively. SDS/PAGE analysis was used for monitoring and identifying proteins in native CSF, depleted CSF samples and the captured fractions. Moreover, shotgun proteomics was used for protein identification in native as well as depleted CSF and the achieved data were compared. Enhanced identification of lower-abundance components was observed in the depleted fraction, in terms of more detected peptides per protein. PMID- 18412541 TI - Characterization of heat-labile uracil-DNA glycosylase from Psychrobacter sp. HJ147 and its application to the polymerase chain reaction. AB - The gene encoding Psp HJ147 UDG (Psychrobacter sp. HJ147 uracil-DNA glycosylase) was cloned and sequenced. The gene consists of 735 bp for coding a protein with 244 amino acid residues. The deduced amino acid sequence of Psp HJ147 UDG showed a high similarity to that of Psychrobacter articus, Psychrobacter cryohalolentis K5 and Psychrobacter sp. PRwf-1. The PCR-amplified Psp HJ147 UDG gene was expressed under the control of the T7lac promoter on pTYB1 in Escherichia coli BL21(DE3). The expressed enzyme was purified with IMPACT-CN (intein-mediated purification with an affinity chitin-binding tag) system. The optimum pH and temperature of the purified enzyme were 7.0-7.5 and 20-25 degrees C respectively. The optimum NaCl and KCl concentrations for the activity of the purified enzyme ranged from 50 to75 mM. The half-life of the enzyme at 50 degrees C was approx. 45 s. These heat-labile characteristics enabled Psp HJ147 UDG to control carry over contamination in direct PCR without loss of the PCR product. Psp HJ147 UDG's contaminant control in both direct PCR and indirect PCR exhibited superiority over the UDG of the marine psychrophilic bacterium strain BMTU 3346 and that of E. coli. PMID- 18412542 TI - Chaperone ligand-discrimination by the TPR-domain protein Tah1. AB - Tah1 [TPR (tetratricopeptide repeat)-containing protein associated with Hsp (heat shock protein) 90] has been identified as a TPR-domain protein. TPR-domain proteins are involved in protein-protein interactions and a number have been characterized that interact either with Hsp70 or Hsp90, but a few can bind both chaperones. Independent studies suggest that Tah1 interacts with Hsp90, but whether it can also interact with Hsp70/Ssa1 has not been investigated. Amino acid-sequence alignments suggest that Tah1 is most similar to the TPR2b domain of Hop (Hsp-organizing protein) which when mutated reduces binding to both Hsp90 and Hsp70. Our alignments suggest that there are three TPR-domain motifs in Tah1, which is consistent with the architecture of the TPR2b domain. In the present study we find that Tah1 is specific for Hsp90, and is able to bind tightly the yeast Hsp90, and the human Hsp90alpha and Hsp90beta proteins, but not the yeast Hsp70 Ssa1 isoform. Tah1 acheives ligand discrimination by favourably binding the methionine residue in the conserved MEEVD motif (Hsp90) and positively discriminating against the first valine residue in the VEEVD motif (Ssa1). In the present study we also show that Tah1 can affect the ATPase activity of Hsp90, in common with some other TPR-domain proteins. PMID- 18412543 TI - Biliverdin reductase is a transporter of haem into the nucleus and is essential for regulation of HO-1 gene expression by haematin. AB - hBVR (human biliverdin reductase) is an enzyme that reduces biliverdin (the product of haem oxygenases HO-1 and HO-2 activity) to the antioxidant bilirubin. It also functions as a kinase and as a transcription factor in the MAPK (mitogen activated protein kinase) signalling cascade. Fluorescence correlation spectroscopy was used to investigate the mobility of hBVR in living cells and its function in the nuclear transport of haematin for induction of HO-1. In transiently transfected HeLa cells only kinase-competent hBVR translocates to the nucleus. A reduced mobility in the nucleus of haematin-treated cells suggests formation of an hBVR-haematin complex and its further association with large nuclear components. The binding of haematin is specific, with the formation of a 1:1 molar complex, and the C-terminal 7-residue fragment KYCCSRK(296) of hBVR contributes to the binding. The following data suggest formation of dynamic complexes of hBVR-haematin with chromatin: (i) the reduction of hBVR mobility in the presence of haematin is greater in heterochromatic regions than in euchromatic domains and (ii) hBVR mobility is not retarded by haematin in nuclear lysates that contain only soluble factors. Moreover, hBVR kinase activity is stimulated in the presence of double-stranded DNA fragments corresponding to HO-1 antioxidant and HREs (hypoxia response elements), as well as by haematin. Experiments with nuclear localization, export signal mutants and si-hBVR [siRNA (small interfering RNA) specific to hBVR] indicate that nuclear localization of hBVR is required for induction of HO-1 by haematin. Because gene regulation is energy-dependent and haematin regulates gene expression, our data suggest that hBVR functions as an essential component of the regulatory mechanisms for haem responsive transcriptional activation. PMID- 18412544 TI - An amino acid at position 142 in nitrilase from Rhodococcus rhodochrous ATCC 33278 determines the substrate specificity for aliphatic and aromatic nitriles. AB - Nitrilase from Rhodococcus rhodochrous ATCC 33278 hydrolyses both aliphatic and aromatic nitriles. Replacing Tyr-142 in the wild-type enzyme with the aromatic amino acid phenylalanine did not alter specificity for either substrate. However, the mutants containing non-polar aliphatic amino acids (alanine, valine and leucine) at position 142 were specific only for aromatic substrates such as benzonitrile, m-tolunitrile and 2-cyanopyridine, and not for aliphatic substrates. These results suggest that the hydrolysis of substrates probably involves the conjugated pi-electron system of the aromatic ring of substrate or Tyr-142 as an electron acceptor. Moreover, the mutants containing charged amino acids such as aspartate, glutamate, arginine and asparagine at position 142 displayed no activity towards any nitrile, possibly owing to the disruption of hydrophobic interactions with substrates. Thus aromaticity of substrate or amino acid at position 142 in R. rhodochrous nitrilase is required for enzyme activity. PMID- 18412545 TI - Similarity and diversity in mechanisms of muscle fate induction between ascidian species. AB - Developmental processes can change during evolution at many levels of the ontogeny of an individual. Embryos of solitary ascidians have a largely invariant mode of development, with fixed cleavage patterns and fate maps. Thus the cell lineages and final body plan of the two quite distantly related species considered in this review, Ciona intestinalis and Halocynthia roretzi, are highly similar. However, close comparison of the developmental mechanisms used by these two species provide examples of evolutionary changes and help pinpoint which aspects of development are evolutionarily flexible. Examples of both similarity and diversity are observed in the mechanisms used to generate the full complement of larval muscle. We will describe the changes in muscle-cell lineage, as well as some striking differences in the intercellular signalling pathways used to induce muscle fate. The somewhat surprising conclusion is that in ascidians, as in nematode vulval development, different signalling mechanisms have been adopted to mediate similar interactions between equivalently positioned cells. PMID- 18412546 TI - Insights into the substrate specificity of plant peptide deformylase, an essential enzyme with potential for the development of novel biotechnology applications in agriculture. AB - The crystal structure of AtPDF1B [Arabidopsis thaliana PDF (peptide deformylase) 1B; EC 3.5.1.88], a plant specific deformylase, has been determined at a resolution of 2.4 A (1 A=0.1 nm). The overall fold of AtPDF1B is similar to other peptide deformylases that have been reported. Evidence from the crystal structure and gel filtration chromatography indicates that AtPDF1B exists as a symmetric dimer. PDF1B is essential in plants and has a preferred substrate specificity towards the PS II (photosystem II) D1 polypeptide. Comparative analysis of AtPDF1B, AtPDF1A, and the type 1B deformylase from Escherichia coli, identifies a number of differences in substrate binding subsites that might account for variations in sequence preference. A model of the N-terminal five amino acids from the D1 polypeptide bound in the active site of AtPDF1B suggests an influence of Tyr(178) as a structural determinant for polypeptide substrate specificity through hydrogen bonding with Thr(2) in the D1 sequence. Kinetic analyses using a polypeptide mimic of the D1 N-terminus was performed on AtPDF1B mutated at Tyr(178) to alanine, phenylalanine or arginine (equivalent residue in AtPDF1A). The results suggest that, whereas Tyr(178) can influence catalytic activity, other residues contribute to the overall preference for the D1 polypeptide. PMID- 18412547 TI - Reduction of S-nitrosoglutathione by alcohol dehydrogenase 3 is facilitated by substrate alcohols via direct cofactor recycling and leads to GSH-controlled formation of glutathione transferase inhibitors. AB - GSNO (S-nitrosoglutathione) is emerging as a key regulator in NO signalling as it is in equilibrium with S-nitrosated proteins. Accordingly, it is of great interest to investigate GSNO metabolism in terms of competitive pathways and redox state. The present study explored ADH3 (alcohol dehydrogenase 3) in its dual function as GSNOR (GSNO reductase) and glutathione-dependent formaldehyde dehydrogenase. The glutathione adduct of formaldehyde, HMGSH (S hydroxymethylglutathione), was oxidized with a k(cat)/K(m) value approx. 10 times the k(cat)/K(m) value of GSNO reduction, as determined by fluorescence spectroscopy. HMGSH oxidation in vitro was greatly accelerated in the presence of GSNO, which was concurrently reduced under cofactor recycling. Hence, considering the high cytosolic NAD(+)/NADH ratio, formaldehyde probably triggers ADH3 mediated GSNO reduction by enzyme-bound cofactor recycling and might result in a decrease in cellular S-NO (S-nitrosothiol) content in vivo. Formaldehyde exposure affected S-NO content in cultured cells with a trend towards decreased levels at concentrations of 1-5 mM, in agreement with the proposed mechanism. Product formation after GSNO reduction to the intermediate semimercaptal responded to GSH/GSNO ratios; ratios up to 2-fold allowed the spontaneous rearrangement to glutathione sulfinamide, whereas 5-fold excess of GSH favoured the interception of the intermediate to form glutathione disulfide. The sulfinamide and its hydrolysis product, glutathione sulfinic acid, inhibited GST (glutathione transferase) activity. Taken together, the findings of the present study provide indirect evidence for formaldehyde as a physiological trigger of GSNO depletion and show that GSNO reduction can result in the formation of GST inhibitors, which, however, is prevented under normal cellular redox conditions. PMID- 18412548 TI - Campylobacter infection of broiler chickens in a free-range environment. AB - Campylobacter jejuni is the most common cause of bacterial gastroenteritis worldwide, with contaminated chicken meat considered to represent a major source of human infection. Biosecurity measures can reduce C. jejuni shedding rates of housed chickens, but the increasing popularity of free-range and organic meat raises the question of whether the welfare benefits of extensive production are compatible with food safety. The widespread assumption that the free-range environment contaminates extensively reared chickens has not been rigorously tested. A year-long survey of 64 free-range broiler flocks reared on two sites in Oxfordshire, UK, combining high-resolution genotyping with behavioural and environmental observations revealed: (i) no evidence of colonization of succeeding flocks by the C. jejuni genotypes shed by preceding flocks, (ii) a high degree of similarity between C. jejuni genotypes from both farm sites, (iii) no association of ranging behaviour with likelihood of Campylobacter shedding, and (iv) higher genetic differentiation between C. jejuni populations from chickens and wild birds on the same farm than between the chicken samples, human disease isolates from the same region and national samples of C. jejuni from chicken meat. PMID- 18412549 TI - Identification of intermediates formed during anaerobic benzene degradation by an iron-reducing enrichment culture. AB - Anaerobic benzene degradation is an important process in contaminated aquifers but is poorly understood due to the scarcity of microbial cultures for study. We have enriched a ferric iron-reducing culture that completely mineralizes benzene to CO(2). With (13)C(6)-labelled benzene as the growth substrate, ring-labelled benzoate was identified as a major intermediate by liquid chromatography/tandem mass spectrometry (LC/MS/MS) analysis of culture supernatants. With increasing incubation time, (13)C(7)-labelled benzoate appeared, indicating that the carboxyl group of benzoate derived from CO(2) that was produced from mineralization of labelled benzene. This was confirmed by growing the culture in (13)C-bicarbonate-buffered medium with unlabelled benzene as the substrate, as the label appeared in the carboxyl group of benzoate produced. Phenol was also identified as an intermediate at high concentration. However, it was clearly shown that phenol was formed abiotically by autoxidation of benzene during the sampling and analysis procedure as a result of exposure to air. The results suggest that, in our culture, anaerobic benzene degradation proceeds via carboxylation and that caution should be exercised in interpreting hydroxylated benzene derivatives as metabolic intermediates of anaerobic benzene degradation. PMID- 18412550 TI - Oxygen-dependent autoaggregation in Shewanella oneidensis MR-1. AB - In aerobic chemostat cultures maintained at 50% dissolved O(2) tension (3.5 mg l( 1) dissolved O(2)), Shewanella oneidensis strain MR-1 rapidly aggregated upon addition of 0.68 mM CaCl(2) and retained this multicellular phenotype at high dilution rates. Confocal microscopy analysis of the extracellular matrix material contributing to the stability of the aggregate structures revealed the presence of extracellular DNA, protein and glycoconjugates. Upon onset of O(2)-limited growth (dissolved O(2) below detection) however, the Ca(2+)-supplemented chemostat cultures of strain MR-1 rapidly disaggregated and grew as motile dispersed cells. Global transcriptome analysis comparing aerobic aggregated to O(2)-limited unaggregated cells identified genes encoding cell-to-cell and cell to-surface adhesion factors whose transcription increased upon exposure to increased O(2) concentrations. The aerobic aggregated cells also revealed increased expression of putative anaerobic electron transfer and homologues of metal reduction genes, including mtrD (SO1782), mtrE (SO1781) and mtrF (SO1780). Our data indicate that mechanisms involved in autoaggregation of MR-1 are dependent on the function of pilD gene which encodes a putative prepilin peptidase. Mutants of S. oneidensis strain MR-1 deficient in PilD and associated pathways, including type IV and Msh pili biogenesis, displayed a moderate increase in sensitivity to H(2)O(2). Taken together, our evidence indicates that aggregate formation in S. oneidensis MR-1 may serve as an alternative or an addition to biochemical detoxification to reduce the oxidative stress associated with production of reactive oxygen species during aerobic metabolism while facilitating the development of hypoxic conditions within the aggregate interior. PMID- 18412551 TI - Diversity, function and evolution of genes coding for putative Ni-containing superoxide dismutases. AB - We examined the phylogenetic distribution, functionality and evolution of the sodN gene family, which has been shown to code for a unique Ni-containing isoform of superoxide dismutase (Ni-SOD) in Streptomyces. Many of the putative sodN sequences retrieved from public domain genomic and metagenomic databases are quite divergent from structurally and functionally characterized Ni-SOD. Structural bioinformatics studies verified that the divergent members of the sodN protein family code for similar three-dimensional structures and identified evolutionarily conserved amino acid residues. Structural and biochemical studies of the N-terminus 'Ni-hook' motif coded for by the putative sodN sequences confirmed both Ni (II) ligating and superoxide dismutase activity. Both environmental and organismal genomes expanded the previously noted phylogenetic distribution of sodN, and the sequences form four well-separated clusters, with multiple subclusters. The phylogenetic distribution of sodN suggests that the gene has been acquired via horizontal gene transfer by numerous organisms of diverse phylogenetic background, including both Eukaryotes and Prokaryotes. The presence of sodN correlates with the genomic absence of the gene coding for Fe SOD, a structurally and evolutionarily distinct isoform of SOD. Given the low levels of Fe found in the marine environment from where many sequences were attained, we suggest that the replacement of Fe-SOD with Ni-SOD may be an evolutionary adaptation to reduce iron requirements. PMID- 18412552 TI - Human skin buffering capacity: an overview. AB - Skin surface acidity can be measured according to two criteria, its value given by pH and its strength determined by the ability of the skin to resist an acidic/alkaline aggression (i.e. acidic/alkaline resistance and neutralization tests). It is the quantitative extent to which the skin resists these changes that defines the term buffer capacity or acid/alkali resistance and neutralization capacity of skin. We review studies investigating the possible mechanisms contributing to the buffering capacity of the epidermis via alkali/acidic aggression tests. This paper discerns which components of the epidermis are most likely responsible for the epidermal buffering ability. PMID- 18412553 TI - An in vivo study of the mechanical properties of facial skin and influence of aging using digital image speckle correlation. AB - BACKGROUND/PURPOSE: The skin on the face is directly attached to the muscle through the superficial musculoaponeurotic system. We show that this can be used to probe skin mechanical properties, in vivo, using, digital image speckle correlation (DISC), a technique that measures the intrinsic cutaneous pore structure displacement following a natural facial deformation. METHODS: We take a series of images, which are then analyzed with DISC to create a displacement vector diagram, from which we can obtain spatially resolved information regarding facial deformation. We then studied the functional form of the displacement as a function of age, location on the face, and skin treatment. Finally, through DISC vector field analysis we investigate the mechanism of wrinkle formation. RESULTS: We first show that facial skin displacement follows the direction of muscular movement and reflects the magnitude of the applied forces. Using DISC vector field analysis, we find that as the skin ages the distribution of forces becomes more condensed, with a marked spatial asymmetry. Analysis of the data, in the perioral region, we find that the skin elasticity decreases exponentially with age, with a decay constant of approximately 32 years. Similar results, but with a larger amplitude, were also found for the periorbital region. Finally, DISC vector field analysis also shows that the location of maximal stress correlated with the location of existing facial wrinkles. CONCLUSION: The DISC method, as a non-contact technique, is a potential clinical research tool for the diagnosis of facial skin condition and underlying muscular activity. We demonstrate how these factors can be used to monitor the effects of aging, formation of wrinkles, and the efficacy of topical applications of skin creams. PMID- 18412554 TI - Evaluation of dark circles of the lower eyelid: comparison between reflectance meters and image processing and involvement of dermal thickness in appearance. AB - BACKGROUND/AIMS: Dark circles of the lower eyelid (DCLE) represent a well-known beauty problem. The pathogenesis of DCLE is obscure, although stasis and hyperpigmentation of the eyelids have been considered to be involved. One reason for the small number of studies on DCLE may be the difficulty in measuring such soft and curved skin as the eyelids using bulky reflectance meters. The purpose of this study was to quantitatively analyze DCLE using various bioengineering methods. METHODS: The lower eyelid and cheek areas of 14 subjects with DCLE and 28 without DCLE were examined using two kinds of reflectance meters and image analysis to measure erythema index (EI), melanin index (MI), and oxygenation index (OX). Ultrasound echo images were also recorded to evaluate the thickness and echo density of the dermis. An in vitro model using collagen gel and hemoglobin solution was also examined as a phantom of eyelid skin. RESULTS: When contact-type reflectance meters were used, no significant differences in EI, MI, and OX were found between groups with and without DCLE. However, mean values of both MI and EI at eyelids were significantly higher in subjects with DCLE on image analysis, paralleling the results of inspection. Mean dermal thickness was significantly smaller in subjects with DCLE. CONCLUSION: Evaluation of EI and MI by image processing methods seems suitable for quantitative evaluation of DCLE, since inadequate contact of the measuring head with the skin is avoided. Whether stasis or hyperpigmentation is more responsible for DCLE remains uncertain. Dermal thickness of eyelid skin may be involved in the appearance of DCLE. PMID- 18412555 TI - Water sorption and electrical properties of a human nail. AB - BACKGROUND/PURPOSE: Water absorption is important for the overall function of keratinized tissues like the stratum corneum and nail. Hence, measurement of water sorption dynamics and water content of these tissues is of great interest. METHODS: We have studied water sorption and electrical properties of a human nail in a measuring cell where the temperature and relative hydration could be controlled. RESULTS: We found the amount of absorbed water to be linearly dependent on ambient relative humidity up to about 70%, followed by a stronger dependency. Furthermore, we found that electrical conductance and capacitance are exponentially dependent on the water content. CONCLUSION: Both electrical conductance and susceptance are good indicators of water content in the nail. The results also possibly indicate that the capacitance is dependent on the mobility of the keratin chains, while the conductance is probably more dependent on water molecule mobility. PMID- 18412556 TI - Biomechanical properties of human hair with different parameters. AB - BACKGROUND/PURPOSE: Investigations on the mechanical properties of human hair are inadequate. This research explores the break force of anagen hair by forming criteria according to the diameter of hair, gender, age, strain, use of drugs and hair dyes and how these criteria affect the mechanical resistance of hair. METHODS: The resistances of anagen hair of 120 volunteers were investigated in our biomechanics laboratory. The relationship between the value of break force of hair and hair diameter, gender, age, strain, use of drugs and hair dyes was examined. Tensile tests were applied to each hair with a 50 N load cell device. RESULTS: The statistical evaluation demonstrated a significant difference between the break forces of hairs with different hair diameters. The hairs with a diameter of 51 mum and above had a significantly higher break force than those of 50 mum and below (P=0.000). An insignificant correlation was observed between break force and strain (P=0.554, r=-0.055) whereas the correlation between diameter and strain was statistically significant (P=0.000, r=0.361). CONCLUSION: The break force of hairs did not show any significant dependency on age, presence of hair dyes, gender and usage of drugs. Yet, a statistically significant difference was observed between the break forces of hairs when compared according to their diameters and strain. More research on the mechanical properties of human hair consistent with biological properties like protein structure, alpha keratin fibers and microfibril-matrix composite behaviors would guide dermatologists for diagnostic symptoms. PMID- 18412557 TI - A nonlinear elastic behavior to identify the mechanical parameters of human skin in vivo. AB - BACKGROUND/PURPOSE: Various analyses have been performed to identify the mechanical properties of the human skin tissue in vivo. They generally use different approaches and hypotheses (behavior laws as well as mechanical tests) and the obtained results are consequently difficult to analyze and compare. In this paper, an inverse method that can be adapted to any kind of mechanical tests and behavior laws is presented. METHOD: A suction deformation performed on the volar aspect of the forearm of a subject is considered. This test is modeled with the finite element method to compare the experimental and simulated curves using an inverse method that allows the skin mechanical parameters identification. This process is based on two optimization algorithms, Kalman's filter and Gauss Newton's methods. To account for the nonlinear behavior of the skin, a specific nonlinear elastic law, which is then compared with standard linear elastic and neo-Hookean's mechanical behaviors, was developed. RESULTS: The obtained results first prove that neither linear elasticity nor neo-Hookean's laws can be used to model the skin. On the contrary, the nonlinear elastic model presents a relevant fit of the experimental curves. The skin thickness is also proved to be another key point to be taken into consideration. CONCLUSIONS: The obtained results are successfully compared with literature and the reliability of the proposed method is underlined with the identification of 300 additional experimental curves. The different works we are currently focusing on are finally introduced. PMID- 18412558 TI - Interfacial kinetics effects on transdermal drug delivery: a computer modeling. AB - BACKGROUND/PURPOSE: Percutaneous permeation is a frequently used approach in drug delivery, but the detailed physics process in the patch--stratum corneum (SC)- viable epidermis system remains unclear: the influence of the interphases in the multilayered structure has been little studied. METHODS: This paper applied the finite-element method to develop a contact algorithm with an interphase element to account for the interphase barrier on drug diffusion and chemical absorption during a transdermal drug delivery process. A more realistic multilayer structure, including the patch, SC and viable epidermis, are incorporated into the algorithm. Both interphases between the patch and SC, and between SC and viable epidermis are considered. RESULTS: Our study confirms that the interphase transfer coefficients have a direct connection with drug concentration and flux distribution along the diffusion paths. The simulation results suggested a potential for the optimal control of drug diffusion. The partition coefficients and other interphase barrier factors can be incorporated into the model. CONCLUSIONS: The algorithm can deal with complicated geometrical conditions, which is difficult using classical analytical approaches. Furthermore, calibrated against experiments, the model may predict more realistically the drug delivery process and drug distribution profiles so as to assist in the patch and even drug design. PMID- 18412559 TI - Reflectance of human skin using colour photometric stereo: with particular application to pigmented lesion analysis. AB - BACKGROUND/PURPOSE: The optical appearance of human skin is highly dependent on the interaction between the illumination (type and position), observer position and the skin surface structure. Different currently available photographic techniques record different aspects of this appearance, each providing its own incomplete description. This limits their usefulness, especially for pigmented skin lesion diagnosis. In this paper a new, easy to use, low-cost photographic method is described,which aims to generate an efficiently encoded yet reasonably complete representation of skin appearance. MATERIAL AND METHODS: A prototype hand-held camera was developed that rapidly acquires six colour images, each with the skin illuminated from a different direction. A novel photometric stereo processing was used to combine these into a colour image of the skin's diffuse reflectance, independent of the skin surface topography, as well as a separate representation of that topography in the form of a surface gradient image. Images of four clinical pigmented skin lesions were evaluated in comparison with conventional digital photographs by both visual judgement and automated lesion boundary detection. RESULTS: The new colour reflectance images were free from the effects of topographical shading, shadowing and specular reflections. Lesion boundaries obtained automatically from the reflectance images were always closer to the outline drawn by a dermatologist than those obtained from conventional photographs. Finally, recombining the colour reflectance and surface gradient data to form a virtual image of the skin surface that is highly realistic in appearance. CONCLUSIONS: The new colour photometric stereo camera produces images of skin and skin tumours in which the reflectance information that is related to subsurface pigment distribution is separated from the surface topographic information. The total information generated by the system, for use in visual or automated analysis, is potentially greater than that for either conventional photography or dermatoscopy alone. Its further development and broader clinical evaluation are warranted to determine its usefulness and role in a wide range of dermatological tasks, including tele-dermatology applications. PMID- 18412560 TI - Relationships between skin properties and environmental parameters. AB - BACKGROUND/PURPOSE: Many authors have written about skin physiological parameters and their changes according to different environmental conditions. Nevertheless, the literature puts in evidence disagreement among different studies due to the great variability in these parameters and due to the difficulty in comparing the results obtained under different working conditions. Hence, the aim of this work is to attempt to clarify the relationship between some skin properties, such as transepidermal water loss (TEWL), skin hydration and mean skin temperature (T(sk)), and the environmental parameters of ambient temperature (T(a)) and relative humidity (RH), with the help of a climatic chamber to make the environment reliable. This work must be considered as the preliminary step of a wider project dealing with textile engineering: the results will be used in identifying criteria for textile design with the aim of producing more comfortable clothing. METHODS: Experiments were carried out in a climatic chamber with independently controlled T(a) and RH. All the combinations between three levels of T(a) (20 degrees C, 25 degrees C and 30 degrees C) and four levels of RH (25%, 45%, 65% and 85%) were used on a panel of six young female subjects. The assessments made were: skin surface hydration using an electrical capacitance method, TEWL using a Tewameter and T(sk) using a set of thermistors. RESULTS: The results showed a significant correlation between TEWL and T(a), while the RH had a weaker effect on TEWL in the temperature range under investigation. Also, T(sk) showed a higher correlation with T(a) compared with RH. Finally, skin surface hydration was found to be strongly affected by both environmental parameters. CONCLUSIONS: The analysis of experimental data resulted in the elaboration of some easy empirical models useful to evaluate the changes in TEWL, skin hydration and T(sk) in different climatic conditions. These relationships must be considered to be valid only in a restricted range of T(a) (20-30 degrees C) and RH (25-85%) for young female subjects (25-35 years old). PMID- 18412561 TI - Alternative evaluation method in vitro for the water-resistant effect of sunscreen products. AB - BACKGROUND/PURPOSE: Sunscreen products today represent a trend of providing not only simple sun protection factor (SPF)/protection of UVA (PFA) but also other additional benefits. For example, as popularized by seasonless use of sunscreens, the special function of water resistance or sand proof is added to sunscreens as well as for leisure. Because a human in vivo test is time consuming and expensive, a screening process has been tried using an accurate in vitro system. In this study, we suggest the development of an in vitro test that can predict the result of in vivo water resistance of sunscreens. METHODS: Water resistance is presented as a comparison of initial SPF and water-exposed SPF by immersion and washing. In order to be comparable with the in vivo test, water immersion and flow were defined as the basic statements. Also, substrate, revolutions per minute (r.p.m.)--rotative velocity--of propeller inducing water flow, and time of immersion were defined as controlled factors. Considering the strength, separation of test material and adhesive texture, a PMMA plate was selected as suitable among commercial substrates: Transpore tape, VITRO SKIN, and PMMA plate. Also, when the PMMA plate was adhered on the wall of a water bath, the water turbulence of the rotational propeller alone was not strong enough to wash off the test material from the substrate. Therefore, PMMA plates were fixed on the axis. In this experiment, the most important thing is whether this in vitro system can predict correctly. Hence, we tried to match the in vitro water resistance following from our control factors and water resistance value of the in vivo test. RESULTS: We found the immersion time and r.p.m. of controlled factors to obtain the target water resistance using design of experiment, MiniTab statistical package. Response optimization yielded the optimal in vitro conditions of 150 r.p.m./60 min. The repeatability and reproducibility of this in vitro system were also good in validation studies. CONCLUSIONS: This study enables to modify an in vivo water resistance test and predict the result of in vivo water resistance by the manufacture of effective equipment and choosing a suitable substrate. Compared with in vivo results, our in vitro system is more time and cost effective, and provides reliable results. PMID- 18412562 TI - Quantitative evaluation of elderly skin based on digital image analysis. AB - BACKGROUND: The evaluation of the skin state when it is healthy at the time of examination, but predisposed to disease, is based solely on the subjective assessment of clinicians. This assessment may vary from moment to moment and from rater to rater. PURPOSE: We focused on skin texture and aimed to develop a new method to evaluate skin integrity of the elderly using a digital skin image, by verifying the reliability of the clinical evaluation of elderly skin image by specialists to create a 'gold standard,' and by analyzing a digital skin image to identify the indices that explain the skin condition as evaluated by specialists. METHODS: A total of 208 skin images were collected from 34 subjects. And then we processed and analyzed images. Modified Kobayashi's method and texture analysis method were used in image analysis. Kobayashi's method included the process of density transformation, cross binarization, short straight line matching and extraction of main sulcus, and calculation of each index. Texture analysis was performed on cross-binarized images for quantification of uniformity. On the other hand, skin textures in images were scored by six plastic surgeons, using a 10-point Likert scale, where 1 represented 'very bad regularity' and 10 represented 'very good regularity.' The inter-rater reliability was verified by means of the intraclass correlation coefficients (ICC). Finally, stepwise multiple regression analysis was used to extract useful indices; where the clinical evaluation of the physicians (gold standard) was considered to be a dependent variable, and indices obtained from digital skin image analysis to be independent variables. RESULTS: The ICC of raters was 0.92 (95% confidence interval; 0.91-0.94) when including all raters, therefore the score of all raters was used. As a result of stepwise multiple regression, the index of interval (L), thickness (T(ave)), energy 0 degrees , and entropy 45 degrees independently explained the clinician evaluation. R(2) in multiple regression equation was 0.59. CONCLUSIONS: It is shown that quantitative evaluation using skin images and their analysis is one method of determining skin integrity in the elderly. The indices to explain the clinical evaluation of specialists were defined and these indices were obtained simply by using a skin image. PMID- 18412563 TI - Fluorescent image analysis for evaluating the condition of facial sebaceous follicles. AB - BACKGROUND/PURPOSE: Conventional methods, such as 'Sebutape' and 'Sebumeter,' can provide quantitative information on sebum excretion but cannot reflect the condition of sebaceous follicles that can be indirectly evaluated with fluorescent colors of sebum. The images of sebum excretion can be obtained with an ultraviolet-A light that is generally called 'Wood's Lamp.' In this study, we describe fluorescent image analysis methods for the detection of sebum and the color segmentation of sebum to evaluate the condition of sebaceous follicles. METHODS/RESULTS: For subject-dependent automatic image analysis, we extracted calibrated image analysis methods that were optimized for digital fluorescent images acquired from our imaging system. The calibrated subjective threshold values for sebum detection were determined by statistically analyzing the number of sebum detected by the automatic threshold value method and the subjective threshold value method (R=0.947, the number of volunteers: 29). In fluorescent color analysis of sebum, the calibrated reference color markers for the red and white colors were extracted with a coefficient of variance of <10%. CONCLUSION: We are confident that such calibrated image analysis methods in combination with our imaging system can provide useful quantitative diagnostic information for sebum-related skin pathologies. PMID- 18412564 TI - Confocal laser microscopic imaging of conspicuous facial pores in vivo: relation between the appearance and the internal structure of skin. AB - BACKGROUND/PURPOSE: Conspicuous facial pores are one of the more serious esthetic defects of most concern to women. Previous microscopic observations of the skin surface around conspicuous pores have discovered large hollows and uneven skin tone. In this study, the observation area was extended from the skin surface to deeper skin to find the characteristic features of conspicuous pores in a wider spectrum. METHODS: First, a magnified surface image of the cheek skin was obtained using a video microscope. Second, replicas were collected from the same area. Third, the horizontal cross-sectioned images of the epidermis and papillary dermis in different depths were non-invasively obtained using in vivo confocal laser scanning microscopy. These images were compared with each other to find a correlation between features of the skin surface and those of deeper layers. RESULTS: In cross-sectioned images of conspicuous pores, a strongly undulated epidermal-dermal junction was commonly observed around a pore's opening. Areas with this feature correlated well to the areas with larger hollows and an uneven skin tone. CONCLUSION: Our results indicate that there is a positive correlation between the incidence of the characteristic feature at the epidermal-dermal junction and the visual appearance of a pore. PMID- 18412565 TI - Influence of climatic conditions on antiperspirant efficacy determined at different test areas. AB - BACKGROUND: The efficacy of antiperspirants is a current topic among the developers of cosmetic products. According to the Food and Drug Administration (FDA) for the US market, efficacy testing performed in the axilla of human volunteers is mandatory. Another method is yet available, which enables comparison of more than one antiperspirant formula in a single study by performing the test on the backs of volunteers. However, how reproducible are these methods, comparing between the back and axilla? Do they differ as a result of seasonal variation? Is a correlation between the results of the two methods possible? METHODS: To answer these questions, the antiperspirant efficacy of aluminium chlorohydrate (ACH) aqueous solutions was investigated in the axilla and on the backs of volunteers, in four separate clinical studies covering cold and warm seasons. Four days of product application were followed by thermal sweat induction on the fifth day, using a sauna. The amount of sweat recovered by weighing cotton pads before and after sweat induction was used to calculate sweat reduction. Testing in the axilla and on the back was performed on the same volunteers simultaneously to achieve the best comparable data. For this reason, the FDA guideline was slightly modified to thermal stimulation in a sauna instead of in a hot room. RESULTS: Increasing concentrations of ACH in aqueous solutions on the backs of volunteers showed a saturation for 8% ACH with a sweat reduction of approximately 50%. The antiperspirant efficacy of solutions containing 4%, 8% or 12% ACH was repeatedly found at the same levels, when tested on the backs during summer, autumn and winter time. Axilla tests, with an 8% ACH aqueous solution, showed strongly varying results for summer and winter time, represented by sweat reduction values of -2% to 25%. As an assumption, these high variations might result from reduced gel formation in cold seasons due to low humidity in the axillae during the application phase. On the back, this effect was avoided by applying occlusive foils after product application. To gain further insight, a study, during which summer conditions were artificially simulated by thermal stimulation during the application phase, again showed decreased antiperspirant efficacy in the axilla for winter conditions with sweat reduction values of 2%, compared with 25% under simulated summer conditions. CONCLUSION: These strongly varying values of sweat reduction in the axilla under summer and winter conditions make comparisons between antiperspirant products difficult and a statement about correlation between the two test sites back and axilla impossible. A standardization of the application phase, comparable to the simulated summer conditions described here, could be a solution to reduce the high variation of results in the axilla. Consequently, testing on the back is not only a more cost-effective method to investigate the antiperspirant efficacy of more than one formulation, but a reproducible method more independent of climatic influences during test implementation than the axilla test method. It could, therefore, be regarded as the method of choice for discriminating antiperspirant efficacy between several products during development of new antiperspirant formulations. PMID- 18412566 TI - Influence of age and hormone replacement therapy on the functional properties of the lips. AB - AIMS: To investigate the influence of age and hormonal status on some functional properties of the lips of women. METHODS: Lips properties were measured and compared through four groups of women (young with normal menses, aged with normal menses, aged and menopaused, aged, menopaused receiving hormonal replacement therapy). The following parameters were recorded: sebum excretion rate on the forehead. On the lower lip: TEWL, mechanical damping, color, tactile acuity. Moreover, capacitance images of each lips were recorded and the mean capacitance measured. RESULTS: Changes in TEWL, mechanical damping and tactile acuity appear clearly linked to age while the increase in lip darkness could be due to hormones. Sebum excretion rate is also clearly linked to hormonal status. Surprisingly, no changes of lips capacitance were detected vs. age or hormonal status. This study confirms that upper lip is more hydrated than the lower one. CONCLUSION: Most of the parameters measured on the lips have similar variations than the same parameters measured on the skin. Only sebum and color appear being dependant on the hormonal status. PMID- 18412567 TI - Identification of melanoma with a gas sensor array. AB - BACKGROUND/PURPOSE: The relationship between diseases and alterations of the airborne chemicals emitted from the body has been found in many different pathologies and in particular for various forms of cancer. Metabolism of cancer cells is greatly altered during their lifetime; then, modification of chemicals is supposed to be large around cancer tissues. Positive hints in this direction were provided, as an example, on studying the breath composition of lung cancer affected subjects. Besides the conventional analytical approaches, in recent years sensor arrays were also applied to these researches considering the chemical composition changes as those occurring in other applications such as for instance, those dealing with food quality measurements. METHODS: In this paper, the first application of sensor arrays to study the differentiation between melanomas and nevi, namely malignant and benign affection of melanocytary cells, respectively, is presented and discussed. The localization of lesions on the skin surface made possible the utilization of differential measurements aimed at capturing the differences between two adjacent skin regions. This approach strongly reduces the influence of skin headspace variability due to the peculiar subjective odour background and the skin odour variability. The measurement campaign involved 40 cases; 10 of these were diagnosed melanomas referred to surgical intervention. Nine of these diagnoses were further confirmed by histological examinations of the removed tissue and one was a false positive. RESULTS: The differences in the chemical composition of headspace were verified with a gas-chromatographic investigation, and the classification of electronic nose data provided an estimated cross-validated accuracy of the same order of magnitude as the currently used diagnostic instruments. CONCLUSION: Electronic nose sensors have been shown to have good sensitivity towards volatile organic compounds emitted by skin lesions, and the method seems to be effective for malign lesions identification. The results presented in this paper encourage a second experimental campaign with a larger number of participants and a systematic use of gas chromatography mass spectrometer technology in order to identify some possible melanoma biomarkers. PMID- 18412568 TI - Changes of skin characteristics during and after local Parafango therapy as used in physiotherapy. AB - BACKGROUND/AIMS: In physiotherapy, fango (mud) application is a frequently used heat therapy. The main therapeutic effects are due to the elevated temperature of the different tissues with a significant redistribution of blood towards the heated area. This may influence several cardiovascular parameters. There is only limited information on the effect of fango application on skin characteristics. It was the aim of the present study to evaluate the effects of fango application on skin temperature, perfusion of the microcirculation and skin colour. At the same time, cardiovascular parameters such as heart rate, systolic blood pressure (SBP) and diastolic blood pressure (DBP) were recorded. METHOD: Eighteen healthy subjects (age 23.7+/-3.8 years) entered the study. The skin characteristics and cardiovascular parameters were measured before, during and after a 21-min fango application at 44.5 degrees C. RESULTS: Skin temperature and perfusion of the microcirculation increased significantly during fango application: from 35.5+/ 0.4 degrees C to 44.3+/-1.2 degrees C for skin temperature and from 23.2+/-8.8 to 197+/-41 p.u. for the skin microcirculation. These two parameters remained elevated during the fango application and decreased slowly to baseline values within 21 min after fango removal. Skin colour (CIELAB, a(*) parameter) increased from 11.0+/-2.5 to 17.9+/-1.9 when comparing pre- with post-treatment values. At the end of the measuring period, the a(*) parameter did not return to baseline values (15.8+/-2.1). Heart rate increased with 8 bpm during the fango therapy and returned to baseline within 3 min after removal of the fango. SBP and DBP varied slightly during the fango application. They returned to baseline values within 21 min after fango removal. CONCLUSION: The skin parameters indicate a transient temperature effect with an increased perfusion of the microcirculation and a flooding of the superficial capacitance system. The cardiovascular parameters were only slightly influenced and remained in the physiological range. Fango application seems not to be too demanding for the cardiovascular system in healthy subjects. PMID- 18412569 TI - Cosmetic effectiveness of topically applied hydrolysed keratin peptides and lipids derived from wool. AB - BACKGROUND/PURPOSE: Skin moisturisation, elasticity, feel and appearance can all be improved through the topical application of protein hydrolysates. Recent studies suggest that supplementing intercellular lipids of the stratum corneum can enhance the functioning of the skin. METHODS: In this study, a hydrolysed keratin peptide (molecular weight <1000 Da) was prepared from wool and tested on skin in two different formulations: an aqueous solution and an internal wool lipids (IWL) liposome suspension. In vivo long-term studies were performed to evaluate the water barrier function of the skin after topical application of different formulations. During the treatment period, hydration and elasticity were determined. A sorption-desorption test was also performed to assess the hygroscopic properties and water-holding capacity of the different treated skin sites. RESULTS: Significant differences were found between the control and treated sites, with the treated areas showing an increase in hydration and elasticity as a result of keratin peptide application. Measurements also indicated that the keratin formulations reinforce the skin barrier integrity, improving its water-holding capacity. CONCLUSION: A combination of the keratin peptide with the IWL showed beneficial effects, indicating that this combination is suitable for designing new cosmetics products. PMID- 18412570 TI - Dental implants: matters of course and controversies. PMID- 18412571 TI - State of the art of oral implants. PMID- 18412572 TI - To save or to extract, that is the question. Natural teeth or dental implants in periodontitis-susceptible patients: clinical decision-making and treatment strategies exemplified with patient case presentations. PMID- 18412573 TI - Implant stability measurements using resonance frequency analysis: biological and biomechanical aspects and clinical implications. PMID- 18412574 TI - The posterior maxilla: clinical considerations and current concepts using Branemark System implants. PMID- 18412575 TI - Immediate implant placement: treatment planning and surgical steps for successful outcome. PMID- 18412576 TI - Immediate/early loading of dental implants. Clinical documentation and presentation of a treatment concept. PMID- 18412577 TI - Soft tissue enhancement around dental implants. PMID- 18412578 TI - Do long-term followed-up Branemark implants commonly show evidence of pathological bone breakdown? A review based on recently published data. PMID- 18412579 TI - Strategies in reconstruction of the atrophic maxilla with autogenous bone grafts and endosseous implants. PMID- 18412580 TI - The use of zygomatic implants for prosthetic rehabilitation of the severely resorbed maxilla. PMID- 18412581 TI - Bone substitutes and growth factors as an alternative/complement to autogenous bone for grafting in implant dentistry. PMID- 18412582 TI - Sinus membrane elevation and simultaneous insertion of dental implants: a new surgical technique in maxillary sinus floor augmentation. PMID- 18412583 TI - Bone morphogenetic proteins and osseointegration: current knowledge - future possibilities. PMID- 18412584 TI - Ceramic abutments and ceramic oral implants. An update. PMID- 18412585 TI - Uveal melanoma: the good and the bad. PMID- 18412586 TI - Who should manage contact lens related microbial keratitis in Australia and New Zealand? PMID- 18412587 TI - Use of an anterior chamber maintainer in the surgical management of traumatic hyphaemas. AB - Surgical intervention in traumatic hyphaemas becomes necessary when high intraocular pressure (IOP) is intractable and there is persistent corneal staining. Anterior chamber (AC) washout and elimination of blood clots have been described previously using one paracentesis. We describe the value of creating two paracenteses. The first one is made in the lower temporal quadrant and accommodates a 20-gauge anterior chamber maintainer (ACM) that is connected to a bottle of Balanced Salt Solution. The second paracentesis is made in an upper quadrant and serves to evacuate liquefied blood and blood clots. With an ACM in place, the fluctuations of intraoperative IOP are minimized and the AC depth is stabilized throughout the operation. The risk of renewed bleeding is reduced because of the continuous positive intraoperative IOP. The ACM is an important tool in the surgical management of traumatic hyphaemas because it facilitates AC washout and reduces iatrogenic damage to the iris and corneal endothelium. PMID- 18412588 TI - Referral pathways and management of contact lens-related microbial keratitis in Australia and New Zealand. AB - BACKGROUND: We examine the referral pathways and treatment for contact lens related microbial keratitis in Australia and New Zealand. METHODS: Cases were reported in May 2003-September 2004; data on presentation, referral and treatment collected from practitioners and via patient interview. Severity was graded, 1 week cure rate estimated, delays in treatment and medications documented. Hospital and private clinic managements were compared. RESULTS: A total of 297 eligible cases were reported; detailed information on treatment and referral pathways was available on a subset of these cases. Presentation was to optometrists (81/200, 41%), general practitioners (GPs) (69/200, 34%) or emergency departments (46/200, 23%). Optometrists referred to private ophthalmologists (47/79, 60%) more often than hospitals (27/79, 34%). GPs initiated treatment (39/68, 57%) but also referred to hospitals (22/68, 32%) and to private ophthalmologists (7/68, 10%). Of all cases, 67% (195/297) were managed in hospitals (29% admitted, 87/297). Hospitalized cases were predominantly managed with fortified aminoglycoside/cephalosporin (66/81, 82%) and others fluoroquinolones (168/195, 86%). Steroids were used in 36% (98/276) commencing on day 5 (median, interquartile range = 3-7). One-week cure rate was 60% (49/82) in private clinics, 72% (62/86) for hospital outpatient cases and 37% (25/67, P < 0.001) for inpatient cases, which were more severe diseases (47%, 52% and 0% mild, respectively). Delays (>/=12 h) receiving therapy were experienced by 33% (55/168) because of initial inappropriate treatment (48/55), time delays (7/55) but not remoteness (P = 0.6). CONCLUSIONS: The majority of treatment is via hospital clinics, but milder disease is managed in private clinics. The referral process via optometrists, GPs and emergency departments is generally efficient; however, one-third of cases experienced some delays before receiving appropriate therapy highlighting the need for timely diagnosis. PMID- 18412589 TI - Risk factors for first presentation of glaucoma with significant visual field loss. AB - PURPOSE: To identify risk factors associated with visual field (VF) loss on first presentation of glaucoma. METHODS: A case-control study of 107 consecutive newly diagnosed glaucoma patients was performed. Information collected included demographic data, reasons for referral, ophthalmological assessment and VF testing. VF grading was based on Advanced Glaucoma Intervention Study (AGIS) scoring and was additionally classified for the presence of a nasal step, paracentral scotoma and/or arcuate scotoma. RESULTS: Demographic parameters were: mean age 59 years (standard deviation [SD] 14), mean deviation -4.37 dB (SD 4.5), intraocular pressure 21.4 mmHg (SD 6.6) and cup:disc ratio 0.69 (SD 0.1). Thirty seven per cent of all participants had a positive family history of glaucoma. Older age was associated with a VF defect at presentation (mean age of 54 years with no VF defect, compared with 63 years for mild, moderate or severe VF defect, P = 0.0014). Multivariate logistic regression analysis identified positive family history (odds ratio [OR] 10.43; 95% confidence interval [CI] 1.67-67.49) and age (OR 1.15; 95% CI 1.06-1.26) as independent risk factors for VF loss at presentation. Fifty-six per cent had a paracentral VF defect at first diagnosis and 29% of patients showed evidence of substantial VF loss with AGIS score of >/=6 on initial presentation. CONCLUSIONS: Patients who have a positive family history of glaucoma are 10 times more likely to have a VF defect at the time of glaucoma diagnosis. An increased focus on family history may help identify these patients earlier and prevent significant visual loss at the time of presentation. PMID- 18412590 TI - Modified Bahasa Malaysia version of VF-14 questionnaire: assessing the impact of glaucoma in rural area of Malaysia. AB - PURPOSE: To evaluate the functional impairment of glaucoma patients, using a modified Bahasa Malaysia version of VF-14 questionnaire, and to correlate the score with the severity of the disease. METHODS: One-to-one interview by trained interviewers was conducted on glaucoma patients seen in the eye clinic of Hospital Universiti Sains Malaysia, using a modified validated Bahasa Malaysia version of VF-14 questionnaire. The severity of glaucoma was determined based on the better-eye Advanced Glaucoma Intervention Study Scale (AGIS) score of visual field analysis on the latest most reliable visual field. The literacy rate, living situation, better-eye visual acuity and lens status were also documented. RESULTS: A total of 110 glaucoma patients were recruited (54.5% primary open angle glaucoma, 21.8% primary angle-closure glaucoma, 19.2% normotensive glaucoma and 4.5% pseudoexfoliative glaucoma) and majority with bilateral involvement. Based on the better-eye AGIS score, 41.5% were in advanced stage, 29.1% moderate and 29.1% mild. There was a significant association between VF-14 scoring with the better-eye AGIS score (r =-0. 579, P < 0.001), age (r = -0.313, P = 0.000) and better-eye visual acuity (r = -0.752, P = 0.000). Based on the multivariate analysis, there was a significant association of the questionnaire score and better-eye AGIS score (P < 0.001). CONCLUSION: The Bahasa Malaysia version of modified VF-14 questionnaire is a useful tool in quantifying quality of life among glaucoma patients in rural area with high illiteracy rate and provides moderate correlation with severity of the disease. Customization of quality of life questionnaire according to custom and culture of the community will provide better insight to the functional impairment of glaucoma patients. PMID- 18412591 TI - Verteporfin photodynamic therapy for anterior segment neovascularization secondary to ischaemic central retinal vein occlusion. AB - PURPOSE: To evaluate the efficacy of photodynamic therapy (PDT) with verteporfin for anterior segment neovascularizations (ASNVs) in patients affected by ischaemic form of central retinal vein occlusion (CRVO). METHODS: Prospective non comparative case series including 10 consecutive patients (10 eyes) affected by ischaemic CRVO. Main outcome measure was the obliteration of ASNV. RESULTS: One month after PDT, biomicroscopic examination showed partial obliteration of iris new vessels and complete closure of angle neovascularization. Iris fluorescein angiography performed 1 week after treatment showed partial closure of the iris new vessels with no evidence of leakage in the late phases. During the subsequent examinations, a partial reopening of the iris and angle new vessels in association with dye leakage on fluorescein angiography was evident. In any case, the fluorescein leakage turned out to be still reduced with respect to the baseline aspects. CONCLUSIONS: Our results show that PDT with verteporfin can partially obliterate ASNVs in eyes affected by ischaemic CRVO preventing from the evolution towards advanced stages of neovascular glaucoma, but is not effective in cases with complete angle synechial closure. PMID- 18412592 TI - The analysis of predicted capsular bag diameter using modified model of capsule measuring ring in Asians. AB - BACKGROUND: This study measured the capsular bag diameter (CBD) of Asian eyes using a modified capsular measuring ring (CMR) after cataract surgery, and identified preoperative predictive factors for postoperative CBD and the change of capsular bag diameter after surgery. METHODS: This prospective study followed 25 eyes undergoing cataract surgery inserted with a CMR before intraocular lens (IOL) implantation at the Department of Ophthalmology, Ilsan Paik Hospital, Inje University, Republic of Korea. Axial length (AL), anterior chamber depth (ACD), lens thickness, corneal power and corneal size were measured preoperatively. CBD was calculated using the CMR in a dilated state at postoperative 1 day, 1 week, 1 month, 2 months, and 1 year. RESULTS: AL (R = 0.713) and ACD (R = 0.402) were found to positively correlate with CBD at postoperative 1 week. AL (P < 0.001) and ACD (P = 0.046) were found to be predictors of CBD. The mean CBD gradually decreased to 11.30 +/- 1.76 mm at 1 day, 10.86 +/- 1.06 mm at 1 week, 10.23 +/- 1.43 mm at 1 month, 9.88 +/- 1.31 mm at 2 months, and 9.85 +/- 1.43 mm at 1 year. CONCLUSION: CBD size could be determined using a CMR implanted during surgery. AL and ACD were predictors of postoperative CBD. Predicting CBD in this way may assist in selecting appropriately sized IOL, which will lead to improved visual quality after cataract surgery. PMID- 18412593 TI - Functional presbyopia in a rural Kenyan population: the unmet presbyopic need. AB - BACKGROUND: Presbyopia is the most common reason for requiring spectacles in low income regions, although the unmet need for presbyopic spectacles in these regions is very high. The aim of this study was to estimate the prevalence of presbyopia, and the functional impairment and spectacle use among persons with presbyopia in a rural Kenyan population. METHODS: A cross-sectional study was carried out in the Rift Valley, Kenya. Clusters were selected through probability proportionate to size sampling, and people aged >/=50 years within the clusters were identified through compact segment sampling. Within the context of this survey, 130 eligible participants were selected for interview and underwent near vision testing. Functional presbyopia was defined as requiring at least +1.00 dioptre in order to read the N8 optotype at a distance of 40 cm in the participant's usual visual state. Participants were corrected to the nearest 0.25 dioptre in order to see N8. Unmet and met presbyopic need, and presbyopic correction coverage were calculated. RESULTS: Functional presbyopia was found in 111 participants (85.4%). Mean age was lower in those with presbyopia (64.1 years vs. 71.5 years, P = 0.004). Increasing degree of addition required to see N8 was significantly associated with increased difficulty with reading (P = 0.04), sewing (P = 0.03), recognizing small objects (P = 0.02) and harvesting grains (P = 0.05). Among participants with functional presbyopia, 5.4% wore reading glasses and 25.2% had prior contact with an eye care professional. The unmet presbyopic need was 80.0%, met presbyopic need was 5.4% and presbyopic correction coverage was 6.3%. Cost was cited as the main barrier to spectacle use in 62% of participants with presbyopia. CONCLUSION: In low-income regions, there is a high prevalence of uncorrected presbyopia, which is associated with near-vision functional impairment. Provision of spectacles for near vision remains a priority in low-income regions. PMID- 18412594 TI - Retinal pigment epithelial tears after intravitreal bevacizumab injection for exudative age-related macular degeneration. AB - PURPOSE: To determine the incidence of and the risk factors for the development of retinal pigment epithelial (RPE) tears after intravitreal bevacizumab (Avastin) injection for the treatment of exudative age-related macular degeneration (AMD). METHODS: A retrospective, multicentre, consecutive interventional case series of all patients with subfoveal exudative AMD treated with intravitreal bevacizumab between August 2005 and April 2007. The main outcome measures were pre- and post-RPE tear visual acuity and choroidal neovascular membrane lesion types, incidence of tears and time from first injection until development of the tear. RESULTS: A total of 920 eyes with exudative AMD were treated with intravitreal bevacizumab. Fifteen eyes from 15 patients developed a RPE tear for an incidence of 1.6%. The average patient age was 79 years. Fourteen of the fifteen eyes (93%) had an occult subfoveal choroidal neovascular membrane. Forty-seven per cent (7/15) of the RPE tears occurred within the first 6 weeks of treatment, and all tears occurred within the first 18 weeks of treatment initiation. The mean pre-injection visual acuity was 20/100 with a mean post-tear visual acuity of 20/200. In all 10 eyes in which the tear involved the fovea, the final visual acuity was poor. Six of the 15 eyes continued with bevacizumab/ranibizumab (Lucentis) injections after tear development, and four of these six eyes continued to have visual improvement. CONCLUSION: RPE tears occur after intravitreal bevacizumab injections for exudative AMD in approximately 1.6% of eyes and can cause severe vision loss. Maintenance of therapy may help preserve vision after RPE tear development. PMID- 18412595 TI - Prevalence of idiopathic macular hole in adult rural and urban south Indian population. AB - BACKGROUND: This study was undertaken to evaluate the prevalence of idiopathic macular hole in a defined community in Southern India. METHODS: In all, 7774 of the 9546 enumerated (81.43%) subjects availed themselves for an ophthalmic assessment which included a detailed ophthalmic examination and fundus photography. All data were entered and stored in a secure computerized database and statistical analysis was performed using spss for Windows. RESULTS: Thirteen subjects comprising six males and seven females were diagnosed with macular holes equating to a risk of 0.17%. Bilateral macular holes were found in two subjects. The mean age of subjects with a macular hole was 67 years. Increasing age and history of cataract surgery was strongly associated with an increased prevalence of macular holes (P < 0.0001). There was no difference in the prevalence of macular hole between the rural and urban communities. The mean logMAR visual acuity of subjects in the eye with a macular hole was 1.31 +/- 0.45 whereas the acuity in the unaffected fellow eye was 0.70 +/- 0.51. The mean spherical equivalent in the subjects with macular hole was -0.56 +/- 1.92 (-4.25 to +1.00). CONCLUSION: Prevalence rate of idiopathic macular hole in South India appears to be comparable to that seen worldwide. PMID- 18412596 TI - Densiron intraocular tamponade: a case series. AB - PURPOSE: To report on the use of Densiron (heavy silicone oil), a new endotamponade agent with proposed benefits in complex retinal detachments involving inferior proliferative vitreoretinopathy. METHOD: Retrospective case note review. RESULTS: Ten cases (10 eyes) in which Densiron was used in eyes which had failed initial retinal detachment surgery. The overall success rate of long-term total reattachment was 70%. Mean preoperative visual acuity was logMAR 2.03 (SD 0.406), which improved to a mean postoperative visual acuity of logMAR 1.00 (SD 0.571) (P = 0.001). CONCLUSION: Densiron was well tolerated with few side-effects and appears to be a useful agent in patients with inferior proliferative vitreoretinopathy. PMID- 18412597 TI - Awareness of diabetic eye disease among general practitioners and diabetic patients in Yangon, Myanmar. AB - BACKGROUND: Diabetes is an increasing problem in Myanmar with more than three million people affected. There are no data on awareness of diabetic retinopathy among the general practitioners (GPs) or diabetic population of Myanmar. This study aims to evaluate the awareness of diabetes-related eye disease among GPs and diabetic patients in Yangon, Myanmar. DESIGN: A cross-sectional survey. METHODS: From the Myanmar Medical Association Registry of 978 practicing GPs in Yangon, 200 were randomly selected and a structured questionnaire was sent to each. Each GP was asked to give a separate questionnaire to the first five diabetic patients who attended their practice. RESULTS: One hundred GPs and 480 patients returned the questionnaires. Although 99% of GPs were aware that diabetes could result in loss of vision, 49% never examined the fundi of their diabetic patients. Of the diabetic patients, 86% were aware that diabetes could damage their eyesight. Although 92% realized they should visit an ophthalmologist regularly, only 57% had seen an ophthalmologist. Patients who never attended school were less likely to visit an ophthalmologist than those with tertiary education (odds ratio 0.24; 95% confidence interval 0.09, 0.66). Patients with diabetes for less than 2 years were less likely to visit an ophthalmologist than those with diabetes for more than 10 years (odds ratio 0.21; 95% confidence interval 0.9, 0.44). There was no association between age, gender or work status and the likelihood of having seen an ophthalmologist. CONCLUSION: Although both GPs and diabetic patients are aware of the need for regular fundal screening, just over half the patients had been screened. There exists a need for programmes in Myanmar to induce a behavioural change in diabetic patients with regards to screening examinations. PMID- 18412598 TI - Eosinophilic angiocentric fibrosis of the orbit. AB - A 30-year-old woman complained of right-sided epiphora for 2 years. She also reported diplopia on certain gaze positions and felt a hard mass behind the right medial inferior orbital rim. Magnetic resonance imaging studies demonstrated a relatively well-delineated mass in the right inferomedial orbit with minimal ethmoid sinus involvement. Histopathological evaluation following a large incisional biopsy showed massive eosinophilic infiltration and fibrosis with the final diagnosis of eosinophilic angiocentric fibrosis. She was then discovered to have significant peripheral eosinophilia and elevated serum IgE levels and clinical findings of allergic rhinitis and sinusitis. She was treated with systemic fluorocortolon and desloratadin for 4 months. She remained stable without recurrence for 32 months. The patient with this exceptionally rare tumour of the orbit benefited from debulking surgery followed by systemic corticosteroids and antihistaminics. PMID- 18412599 TI - The hunt for the secrets of uveal melanoma. AB - The many secrets of uveal melanoma are being uncovered. Information on host and environmental factors that predispose to uveal melanoma has been published. The most important factors include light eye colour, fair skin, inability to tan and chronic sun exposure. Clinical clues that are visible on ophthalmoscopy have been shown to be significant factors in predicting growth of small borderline tumours and allow for early detection of melanoma. These factors include thickness over 2 mm, subretinal fluid, symptoms, orange pigment overlying the tumour and tumour margin within 3 mm of the disc. Refined methods of cytogenetic analysis have identified several chromosomal mutations associated with uveal melanoma. Currently, the most important mutation proves to be chromosome 3 monosomy, an abnormality associated with greater risk for metastatic disease. PMID- 18412600 TI - Meta-analysis of randomized controlled trials comparing timolol with brimonidine in the treatment of glaucoma. AB - This paper aims to compare the efficacy and tolerability of timolol versus brimonidine in the treatment of glaucoma. Comprehensive searches were performed using Medline, Embase and the Cochrane Controlled Trials Register for randomized controlled trials comparing timolol and brimonidine. Two reviewers independently assessed trials for eligibility and quality and extracted data. A random effects model was used to combine studies. Outcome was defined as the absolute mean intraocular pressure (IOP) reduction from baseline to end-point for efficacy, and relative risk (RR) for adverse events. Subgroup analysis and meta-regression were used to explore heterogeneity according to trial design and quality. Ten publications reporting on eight trials with 2387 participants were included in the meta-analysis. Two further trials were commented on qualitatively. IOP reduction was not significantly different between timolol and brimonidine. Weighted mean difference (WMD) of IOP reduction was 0.24 mmHg (favouring brimonidine) with a 95% confidence interval of -0.57 to 1.04 mmHg. There was significant heterogeneity between studies (chi(2) (13) = 73.75, P < 0.00001, I(2) = 91%). Subgroup analysis showed no significant WMD for studies where data were analysed from end-points >/=6 months or <6 months. Meta-regression analysis showed increased WMD IOP reduction in favour of brimonidine with increased trial quality (t(3) = -4.58, P = 0.01), but no significant association with trial duration (t(3) = 0.73, P = 0.51) or size (t(3) = -0.59, P = 0.57). The RR of ocular allergy was much lower with timolol than brimonidine (RR = 0.08, 95% confidence interval 0.01 to 0.47). Publication bias was not evident on a funnel plot, although the number of studies was small. The conclusion is that both drugs are equally effective in lowering IOP. Brimonidine is associated with a higher rate of allergy. PMID- 18412601 TI - Reverse horseshoe tear morphology for traumatic retinal dialysis. PMID- 18412602 TI - Benign paroxysmal tonic upgaze of childhood in a hypotonic infant with Beckwith Wiedemann syndrome. PMID- 18412603 TI - Intravitreal bevacizumab for choroidal neovascularisation secondary to punctate inner choroidopathy. PMID- 18412604 TI - A piece of my mind. PMID- 18412605 TI - Similar to acne vulgaris, bacteria may produce the biological glue that causes plugging of the meibomian gland leading to chalazions. PMID- 18412607 TI - Helicobacter pylori infection as an environmental familial clustering risk factor for primary open-angle glaucoma. PMID- 18412610 TI - Regulation of neurofilament dynamics by phosphorylation. AB - Neurofilament (NF) phosphorylation has long been considered to regulate axonal transport rate and in doing so to provide stability to mature axons. Studies utilizing mice in which the C-terminal region of NF subunits (which contains the vast majority of phosphorylation sites) has been deleted has prompted an ongoing challenge to this hypothesis. We evaluate the collective evidence to date for and against a role for NF C-terminal phosphorylation in regulation of axonal transport and in providing structural support for axons, including some novel studies from our laboratory. We present a few suggestions for further experimentation in this area, and expand upon previous models for axonal NF dynamics. Finally, we address how C-terminal phosphorylation is regionally and temporally regulated by a balance of kinase and phosphatase activities, and how misregulation of this balance can contribute to motor neuron disease. PMID- 18412611 TI - The role of the subthalamic nucleus in 'compulsive' behavior in rats. AB - Different lines of evidence point to dysfunction of basal ganglia-thalamocortical circuits in obsessive-compulsive disorder (OCD). It has been hypothesized that the circuits' dysfunction in OCD may be characterized by a relative under activity of the indirect compared with the direct pathway within these circuits. The present study tested whether lesions of the subthalamic nucleus (STN), a major node of the indirect pathway, would affect compulsive behavior, using the signal attenuation rat model of OCD. In this model, compulsive lever-pressing is induced by the attenuation of an external signal of reward delivery; an attenuation that is hypothesized to simulate the deficient response feedback suggested to underlie obsessions and compulsions in patients with OCD. Rats sustaining lesions to the STN showed a selective increase in compulsive lever pressing compared with sham-operated rats. A post mortem biochemical analysis revealed a decrease in serotonin content in the prelimbic and infralimbic cortices, caudate-putamen (but not nucleus accumbens), globus pallidus and substantia nigra-ventral tegmental area, as well as a decrease in dopamine content in the caudate-putamen in STN-lesioned compared with sham rats. A comparison to recent findings that lesions to the orbitofrontal cortex, which also result in a selective increase in compulsive lever-pressing, lead to a decrease in serotonin and dopamine content in the caudate-putamen suggests that there may be a final common pathway by which different brain pathologies may lead to a pro-compulsive state. PMID- 18412612 TI - Neuroplasticity in brain reward circuitry following a history of ethanol dependence. AB - Mitogen-activated and extracellular regulated kinase (MEK) and extracellular signal-regulated protein kinase (ERK) pathways may underlie ethanol-induced neuroplasticity. Here, we used the MEK inhibitor 1,4-diamino-2,3-dicyano-1,4 bis(2-aminophenylthio)butadiene (UO126) to probe the role of MEK/ERK signaling for the cellular response to an acute ethanol challenge in rats with or without a history of ethanol dependence. Ethanol (1.5 g/kg, i.p.) induced expression of the marker genes c-fos and egr-1 in brain regions associated with both rewarding and stressful ethanol actions. Under non-dependent conditions, ethanol-induced c-fos expression was generally not affected by MEK inhibition, with the exception of the medial amygdala (MeA). In contrast, following a history of dependence, a markedly suppressed c-fos response to acute ethanol was found in the medial pre frontal/orbitofrontal cortex (OFC), nucleus accumbens shell (AcbSh) and paraventricular nucleus (PVN). The suppressed ethanol response in the OFC and AcbSh, key regions involved in ethanol preference and seeking, was restored by pre-treatment with UO126, demonstrating a recruitment of an ERK/MEK-mediated inhibitory regulation in the post-dependent state. Conversely, in brain areas involved in stress responses (MeA and PVN), an MEK/ERK-mediated cellular activation by acute ethanol was lost following a history of dependence. These data reveal region-specific neuroadaptations encompassing the MEK/ERK pathway in ethanol dependence. Recruitment of MEK/ERK-mediated suppression of the ethanol response in the OFC and AcbSh may reflect devaluation of ethanol as a reinforcer, whereas loss of an MEK/ERK-mediated response in the MeA and PVN may reflect tolerance to its aversive actions. These two neuroadaptations could act in concert to facilitate progression into ethanol dependence. PMID- 18412613 TI - Genetic enhancement of trace fear memory and cingulate potentiation in mice overexpressing Ca2+/calmodulin-dependent protein kinase IV. AB - Long-term potentiation (LTP) is a key cellular model for studying mechanisms for learning and memory. Previous studies reported that the Ca(2+)/calmodulin dependent protein kinase IV (CaMKIV) is critical for gene regulation, and behavioral learning and memory. Less is known about the roles of CaMKIV in cortical plasticity and trace fear memory. Here we have found that LTP was significantly enhanced in the anterior cingulate cortex (ACC) of the mice overexpressing CaMKIV. By contrast, neither alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionic acid (AMPA) receptor-mediated basal excitatory synaptic transmission nor N-methyl-d-aspartate (NMDA) receptor-mediated excitatory postsynaptic currents were affected. Furthermore, paired-pulse ratio in the transgenic mice is normal. In behavioral tests, we found that the CaMKIV transgenic mice exhibited significant enhancement in trace fear memory, while the acute sensory thresholds were not affected. Our results provide strong evidence that forebrain CaMKIV contributes to trace fear memory by enhancing synaptic potentiation in the ACC. PMID- 18412614 TI - Unstable CA1 place cell representation in rats with entorhinal cortex lesions. AB - Recent studies emphasize the importance of the entorhinal cortex in spatial representation and navigation. Furthermore, evidence is accumulating to show that spatial processing depends on interactions between the entorhinal cortex and the hippocampus. To investigate these interactions, we examined the effects of entorhinal cortex lesions on the activity of hippocampal CA1 place cells. Rats received bilateral radiofrequency lesions of the entorhinal cortex or sham lesions before place cell recording. Place cells were recorded as the rats performed a pellet-chasing task in a cylinder containing three cue-objects. Entorhinal cortex lesions did not abolish place cell spatial firing but reduced noticeably discharge rate and field size. Most importantly, the lesions affected firing field stability when cells were recorded both in constant conditions and following cue manipulations (object rotation, object removal). These findings indicate that the entorhinal cortex is necessary for the stability of hippocampal representations across exposures to a familiar environment. Consistent with the recent discovery of grid cells in the medial entorhinal cortex, our results suggest that the entorhinal cortex contributes to providing a spatial framework that would enable the hippocampus to maintain stable environment-specific representations. PMID- 18412615 TI - Oxytocin reduces anxiety via ERK1/2 activation: local effect within the rat hypothalamic paraventricular nucleus. AB - The neuropeptide oxytocin (OT) modulates social behaviours and is an important anxiolytic substance of the brain. However, sites of action and the intracellular signalling pathways downstream of OT receptors (OTR) within the brain remain largely unknown. In the present studies, we localized the anxiolytic effect of OT by bilateral microinfusion of OT (0.01 nmol/0.5 microL) into the hypothalamic paraventricular nucleus (PVN) in male rats using both the elevated plus-maze and the light-dark box. Moreover, intracerebroventricular administration of OT, but not of the related neuropeptide vasopressin (VP), dose-dependently activated the extracellular signal-regulated kinase 1/2 (ERK1/2) cascade. Specifically, OT induced the phosphorylation of Raf-1, MEK1/2 and ERK1/2 in the hypothalamus in vivo and in hypothalamic H32 neurons via EGF receptors. OT-induced ERK1/2 phosphorylation was immunohistochemically localized within VP neurons of the PVN and the supraoptic nucleus. Importantly, the anxiolytic effect of OT within the PVN was prevented by local inhibition of the MAP kinase cascade with a MEK1/2 inhibitor (U0126, 0.5 nmol/0.5 microL) locally infused prior to OT, indicating the causal involvement of this intracellular signalling cascade in the behavioural effect of OT. OT effects within the hypothalamus may have far reaching implications for the regulation of emotionality and social behaviours and, consequently, for the development of possible therapeutic strategies to treat affective disorders. Thus, OTR agonism or activation of the ERK1/2 cascade, specifically within the hypothalamus, may provide therapeutically relevant mechanisms. PMID- 18412616 TI - Long-term potentiation in the nucleus accumbens requires both NR2A- and NR2B containing N-methyl-D-aspartate receptors. AB - N-methyl-d-aspartate (NMDA) receptors play crucial roles in several forms of long term changes in the efficacy of glutamatergic synaptic transmission. The suggestion that the NR2A subunit of the NMDA receptor may be selectively involved in the induction of long-term potentiation (LTP) in the hippocampus and cortex has been challenged. However, the contribution of NR2B in the induction of LTP is not always clearly established. The present study investigates the role of NR2A and NR2B in the induction of LTP in the nucleus accumbens (NAc), a brain region that expresses high levels of NR2B and an NMDA-dependent form of LTP. We recorded extracellular field excitatory postsynaptic potentials/population spikes in slices of mouse NAc. High-frequency stimulation of glutamatergic fibers consistently induced LTP of the field excitatory postsynaptic potential/population spike in the NAc. LTP was abolished in the presence of selective antagonists of either NR2B [R-(R*,S*)-alpha-(4-hydroxyphenyl)-beta methyl-4-(phenyl-methyl)-1-piperidine propanol and Ifenprodil] or NR2A ([(R)-[(S) 1-(4-bromo-phenyl)-ethylamino]-(2,3-dioxo-1,2,3,4-tetrahydroquinoxalin-5-yl) methyl]-phosphonic acid) subunits. Recordings performed in a low concentration of Mg(2+) ions in the perfusion solution did not reveal a selective involvement of a particular NMDA receptor subunit because either NR2A or NR2B antagonists were able to block LTP. LTP was also abolished in the presence of a low concentration of the non-subunit-selective NMDA receptor antagonist dl-2-amino-5 phosphonopentanoic acid in normal Mg(2+) and low Mg(2+) in the perfusion solution. These results show that the degree of NMDA receptor activation, and not their subunit composition, determines whether LTP is induced in the NAc. PMID- 18412617 TI - A circulating ghrelin mimetic attenuates light-induced phase delay of mice and light-induced Fos expression in the suprachiasmatic nucleus of rats. AB - Anatomical evidence suggests that the ventromedial arcuate nucleus (vmARC) is a route for circulating hormonal communications to the suprachiasmatic nucleus (SCN). Whether this vmARC-SCN connection is involved in the modulation of circadian activity of the SCN is not yet known. We recently demonstrated, in rats, that intravenous (i.v.) injection of a ghrelin mimetic, GHRP-6, during the daytime activated neurons in the vmARC and reduced the normal endogenous daytime Fos expression in the SCN. In the present study we show that i.v. administration of GHRP-6 decreases light-induced Fos expression at ZT13 in the rat SCN by 50%, indicating that light-induced changes in the SCN Fos expression can also be reduced by GHRP-6. Because it is difficult to study light-induced phase changes in rats, we examined the functional effects of GHRP-6 on light-induced phase shifts in mice and demonstrated that peripherally injected GHRP-6 attenuates light-induced phase delays at ZT13 by 45%. However, light-induced Fos expression in the mice SCN was not blocked by GHRP-6. These results illustrate that acute stimulation of the ghrelinergic system may modulate SCN activity, but that its effect on light-induced phase shifts and Fos expression in the SCN might be species related. PMID- 18412618 TI - Classical and melanopsin photoreception in irradiance detection: negative masking of locomotor activity by light. AB - Studies in mice lacking either classical or melanopsin photoreception have been useful in describing the photoreceptor contribution to irradiance detection in accessory visual responses. However, application of these findings to irradiance detection in intact animals is problematical because retinal degeneration or manipulation can induce secondary changes in the retina. Among responses dependent on irradiance detection, the suppression of activity by light (negative masking) has had limited study. To further understand the function of classical and melanopsin photoreceptors we studied irradiance and spectral sensitivity of masking by light, primarily in mice with intact retinae. The sensitivity of negative masking was equivalent for medium ( approximately 500 nm) and short wavelengths ( approximately 365 nm) in three strains of wild-type mice, identifying a marked short-wavelength-sensitive-cone input. At medium wavelengths, spectral sensitivity above 500 nm had closest fit to the nomogram for the medium-wavelength-sensitive-cone, but a combined input of cone and melanopsin photoreceptors in wild-type mice seems likely. Under white light a decompression of the irradiance range of masking in C3H rd/rd cl mice, lacking rods and cones, identified a functional deficiency presumably resulting from the absence of classical photoreceptor input. Together the evidence demonstrates a pronounced and sustained classical photoreceptor input to irradiance detection for negative masking, and suggests one role of classical photoreceptor input is to constrain dynamic range. PMID- 18412619 TI - Changes in Fos expression in the accessory olfactory bulb of sexually experienced male rats after exposure to female urinary pheromones. AB - We studied Fos-immunoreactive (Fos-ir) structures in the accessory olfactory bulb (AOB) of rats after the vomeronasal organ was exposed to urine. Exposure of the vomeronasal organ of male Wistar rats to oestrous and dioestrous female Wistar urine led to the appearance of many Fos-ir cells in the rostral region of the periglomerular cell (PGC) layer, but induced few Fos-ir cells in the caudal region. These results suggest that the regionalization of Fos-ir cells after exposure to female urine is remarkable in the PGC layer of the AOB. Sexually experienced male rats have been shown to prefer oestrous to dioestrous female urine, while sexually inexperienced males do not exhibit these preferences. In the present study, we compared the expression of Fos-ir cells in the AOB of sexually experienced and sexually inexperienced male rats following exposure to oestrous and dioestrous urine. In the localized region (lateral and rostral sectors) of the PGC layer, many more Fos-ir cells were expressed in the sexually experienced rats than in the inexperienced rats. These results suggest that sexual experience in males enhances the transmission of reproductively salient information concerning potential oestrous status to a specific PGC region of the AOB. PMID- 18412620 TI - Osmotic modulation of stimulus-evoked responses in the rat supraoptic nucleus. AB - Neural information is conveyed by action potentials along axons to downstream synaptic targets. Synapses permit functionally relevant modulation of the information transmitted by converging inputs. Previous studies have measured the amount of information associated with a given stimulus based either on spike counts or on the relative frequencies of spike sequences represented as binary strings. Here we apply information theory to the phase-interval stimulus histogram (PhISH) to measure the extent of the stimulus-evoked response using the statistical relationship between each interspike interval and its phase within the stimulus cycle. We used the PhISH as a novel approach to investigate how different osmotic states affect the flow of information through the osmoreceptor complex of the hypothalamus. The amount of information conveyed from one (afferent) element of the complex, the anteroventral region of the third ventricle (AV3V), to another (an efferent element), the supraoptic nucleus, was increased by hypertonic stimulation (intravenous mannitol, z = 4.39, P < 0.001) and decreased by hypotonic stimulation (intragastric water, z = -3.37, P < 0.001). Supraoptic responses to AV3V stimulation differed from those that follow stimulation of a hypothalamic element outside the osmoreceptor complex, the suprachiasmatic nucleus (SCN), which also projects to the supraoptic nucleus. Thus osmosensitive gain control mechanisms differentially modulate osmotically dependent and osmotically independent inputs, and enhance the osmoresponsiveness of supraoptic cells within a physiological range. The value of the novel approach is that its use is not limited to the osmoreceptor ensemble but it can be used to investigate the flow of information throughout the central nervous system. PMID- 18412622 TI - Balance training and ballistic strength training are associated with task specific corticospinal adaptations. AB - The aim of this study was to investigate the role of presumably direct corticospinal pathways in long-term training of the lower limb in humans. It was hypothesized that corticospinal projections are affected in a training-specific manner. To assess specificity, balance training was compared to training of explosive strength of the shank muscles and to a nontraining group. Both trainings comprised 16 1-h sessions within 4 weeks. Before and after training, the maximum rate of force development was monitored to display changes in motor performance. Neurophysiological assessment was performed during rest and two active tasks, each of which was similar to one type of training. Hence, both training groups were tested in a trained and a nontrained task. H-reflexes in soleus (SOL) muscle were tested in order to detect changes at the spinal level. Corticospinal adaptations were assessed by colliding subthreshold transcranial magnetic stimulation to condition the SOL H-reflex. The short-latency facilitation of the conditioned H-reflex was diminished in the trained task and enhanced in the nontrained task. This was observable in the active state only. On a functional level, training increased the rate of force development suggesting that corticospinal projections play a role in adaptation of leg motor control. In conclusion, long-term training of shank muscles affected fast corticospinal projections. The significant interaction of task and training indicates context specificity of training effects. The findings suggest reduced motor cortical influence during the trained task but involvement of direct corticospinal control for new leg motor tasks in humans. PMID- 18412621 TI - Bilateral coordination of walking and freezing of gait in Parkinson's disease. AB - Freezing of gait (FOG) in Parkinson's disease (PD) occurs most frequently during turns or step initiation, two tasks that likely demand a high degree of bilateral coordination between the legs. Our objective was to test the hypothesis that impairments in bilateral coordination of stepping are associated with FOG in PD. We compared locomotion features while walking on level ground between patients with PD that experience FOG (PD + FOG; n = 21) and patients with PD that do not (PD - FOG; n = 13). To study bilateral stepping coordination, we defined the stride duration of one foot as a gait cycle or 360 degrees , determined the relative timing of contralateral heel-strikes and defined this as the phase, phi (ideally, phi = 180 degrees ). The sum of the coefficient of variation of phi and the mean absolute difference between phi and 180 degrees was defined as the phase coordination index (PCI), representing variability and inaccuracy, respectively, in phase generation. During the 'Off' state (= 12 h off anti-parkinsonian medication), PCI values were higher (poorer coordination) in PD + FOG compared with PD - FOG (P < 0.024). Stride-to-stride phase adjustments, Deltaphi, were also studied. Both groups scaled their 'converging' adjustments (towards 180 degrees ) to the same extent, but when generating diverging Deltaphi (away from 180 degrees ), PD + FOG patients exhibited larger errors compared with PD - FOG patients (P < 0.006). This study demonstrates that patients with PD who experience FOG have distinctive impairments in the bilateral coordination of locomotion. Poor bilateral coordination of walking may predispose to FOG, especially during challenging tasks that demand a high degree of left-right coordination. PMID- 18412623 TI - Insulin increases excitability via a dose-dependent dual inhibition of voltage activated K+ currents in differentiated N1E-115 neuroblastoma cells. AB - A role in the control of excitability has been attributed to insulin via modulation of potassium (K(+)) currents. To investigate insulin modulatory effects on voltage-activated potassium currents in a neuronal cell line with origin in the sympathetic system, we performed whole-cell voltage-clamp recordings in differentiated N1E-115 neuroblastoma cells. Two main voltage activated K(+) currents were identified: (a) a relatively fast inactivating current (I(fast) - time constant 50-300 ms); (b) a slow delayed rectifying K(+) current (I(slow) - time constant 1-4 s). The kinetics of inactivation of I(fast), rather than I(slow), showed clear voltage dependence. I(fast) and I(slow) exhibited different activation and inactivation dependence for voltage, and have different but nevertheless high sensitivities to tetraethylammonium, 4 aminopyridine and quinidine. In differentiated cells - rather than in non differentiated cells - application of up to 300 nm insulin reduced I(slow) only (IC(50) = 6.7 nm), whereas at higher concentrations I(fast) was also affected (IC(50) = 7.7 microm). The insulin inhibitory effect is not due to a change in the activation or inactivation current-voltage profiles, and the time-dependent inactivation is also not altered; this is not likely to be a result of activation of the insulin-growth-factor-1 (IGF1) receptors, as application of IGF1 did not result in significant current alteration. Results suggest that the current sensitive to low concentrations of insulin is mediated by erg-like channels. Similar observations concerning the insulin inhibitory effect on slow voltage activated K(+) currents were also made in isolated rat hippocampal pyramidal neurons, suggesting a widespread neuromodulator role of insulin on K(+) channels. PMID- 18412624 TI - Serotonin modulates transmitter release at central Lymnaea synapses through a G protein-coupled and cAMP-mediated pathway. AB - Neuromodulation is central to all nervous system function, although the precise mechanisms by which neurotransmitters affect synaptic efficacy between central neurons remain to be fully elucidated. In this study, we examined the neuromodulatory action of serotonin [5-hydroxytryptamine (5-HT)] at central synapses between identified neurons from the pond snail Lymnaea stagnalis. Using whole-cell voltage-clamp and sharp electrode recording, we show that 5-HT strongly depresses synaptic strength between cultured, cholinergic neuron visceral dorsal 4 (VD4 - presynaptic) and its serotonergic target left pedal dorsal 1 (LPeD1 - postsynaptic). This inhibition was accompanied by a reduction in synaptic depression, but had no effect on postsynaptic input resistance, indicating a presynaptic origin. In addition, serotonin inhibited the presynaptic calcium current (I(Ca)) on a similar time course as the change in synaptic transmission. Introduction of a non-condensable GDP analog, GDP-beta-S, through the presynaptic pipette inhibited the serotonin-mediated effect on I(Ca.) Similar results were obtained with a membrane-impermeable inactive cAMP analog, 8OH-cAMP. Furthermore, stimulation of the serotonergic postsynaptic cell also inhibited presynaptic currents, indicating the presence of a negative feedback loop between LPeD1 and VD4. Taken together, this study provides direct evidence for a negative feedback mechanism, whereby the activity of a presynaptic respiratory central pattern-generating neuron is regulated by its postsynaptic target cell. We demonstrate that either serotonin or LPeD1 activity-induced depression of presynaptic transmitter release from VD4 involves voltage-gated calcium channels and is mediated through a G-protein-coupled and cAMP-mediated system. PMID- 18412625 TI - Postsynaptic expression of an epidermal growth factor receptor regulates cholinergic synapse formation between identified molluscan neurons. AB - Epidermal growth factor (EGF) family members are conserved in both vertebrates and invertebrates. Recent studies suggest that EGF ligands in invertebrates may have neurotrophic actions that possibly compensate for the apparent absence of neurotrophins in these species. In this study, we have cloned an EGF receptor from the mollusk Lymnaea stagnalis (L-EGFR), and shown that L-EGFR is the receptor for a previously identified EGF-like peptide in Lymnaea, named Lymnaea EGF (L-EGF). Knock-down of L-EGFR expression prevented L-EGF-induced excitatory synapse formation between identified cholinergic neuron visceral dorsal 4 (VD4) and its postsynaptic partner left pedal dorsal 1 (LPeD1). Moreover, knock-down of L-EGFR also prevented synapse formation induced by Lymnaea brain conditioned medium, suggesting that L-EGF is the most important, if not the only, brain derived factor that promotes excitatory cholinergic synapse formation in Lymnaea. Thus, our data establish canonical EGF/EGFR signaling as an important synaptotrophic mechanism in invertebrates. PMID- 18412626 TI - Cortical spreading depression modulates synaptic transmission of the rat lateral amygdala. AB - Clinical and pathophysiological evidence connects migraine and the amygdala. Cortical spreading depression (CSD) plays a causative role in the generation of aura symptoms. However, the role of CSD in the pathophysiology of other symptoms of migraine needs to be investigated. An in vitro brain slice technique was used to investigate CSD effects on tetanus-induced long-term potentiation (LTP) in the lateral amygdala (LA) of the combined rat amygdala-hippocampus-cortex slices. More than 75% of CSD induced in temporal cortex propagated to LA. Induction of CSD in combined amygdala-hippocampus-cortex slices in which CSD propagated from neocortex to LA significantly augmented LTP in LA. LTP was inhibited when CSD travelled only in the neocortical tissues. Separation of the amygdala from the remaining neocortical part of the slice, in which CSD propagation was limited to the neocortex, increased LTP close to the control levels. Pharmacological manipulations of the slices, in which CSD reached LA, revealed the involvement of NMDA and AMPA glutamate subreceptors as well as dopamine D2 receptors in the enhancement of LTP in LA. However, neither blocking of GABA receptors nor activation of dopamine D1 receptors affected LTP in these slices. The results indicate the disturbances of LA synaptic transmission triggered by propagation of CSD. This perturbation of LA synaptic transmission induced by CSD may relate to some symptoms occurring during migraine attacks. PMID- 18412627 TI - Synaptic sodium spikes trigger long-lasting depolarizations and slow calcium entry in rat olfactory bulb granule cells. AB - In the mammalian olfactory bulb, axonless granule cells mediate self- and lateral inhibitory interactions between mitral/tufted cells via reciprocal dendrodendritic synapses. Synaptic output from granule cells occurs on both fast and slow timescales, allowing for multiple granule cell functions during olfactory processing. We find that granule cell sodium action potentials evoked by synaptic activation of the sensory input via mitral/tufted cells are followed by a long-lasting depolarization that is not observed after current-evoked action potentials or large excitatory postsynaptic potentials in the same cell. Using two-photon imaging in acute rat brain slices, we demonstrate that this prolonged electrical response is paralleled by an unusual, long-lasting postsynaptic calcium signal. We find that this slow synaptic Ca(2+) signal requires sequential activation of NMDA receptors, a nonselective cation conductance I(CAN) and T-type voltage-dependent Ca(2+) channels. Remarkably, T-type Ca(2+) channels are of critical importance for the 'globalization' of Ca(2+) transients. In individual active spines, the local synaptic Ca(2+) signal summates at least linearly with the global spike-mediated Ca(2+) signal. We suggest that this robust slow synaptic Ca(2+) signal triggers dendritic transmitter release and thus contributes to slow synaptic output of the granule cell. Therefore, the synaptic sodium spike signal could represent a special adaptation of granule cells to the wide range of temporal requirements for their dendritic output. Our findings demonstrate with respect to neuronal communication in general that action potentials evoked by somatic current injection may lack some of the information content of 'true' synaptically evoked spikes. PMID- 18412628 TI - Local oscillations of spiking activity in organotypic spinal cord slice cultures. AB - The origin of rhythm generation in mammalian spinal cord networks is still poorly understood. We have previously proposed that disinhibition-induced rhythms are based on intrinsic firing, recurrent excitation and several mechanisms to de activate the network. In order to clarify these mechanisms we here investigated spontaneous spike discharge oscillations in rat spinal cord slice cultures using multi-electrode arrays and patch clamp. Episodes of such oscillations at 8.5 Hz spontaneously appeared in the ventral parts of the cultured slices. The rising phase of their initial cycles was entirely based on AMPA/kainate receptor dependent recurrent excitation. Initial oscillations were changed into persistent activity by bicuculline and other blockers of GABA A, but not by blockers of glycine receptors, suggesting a role for GABAergic synaptic inhibition in network de-activation during oscillation cycles. Blockade of glycine receptors by strychnine caused a prolongation of oscillations and their spreading in the slice, suggesting that these receptors are mainly involved in the spatial and temporal restriction of oscillations. In most cultures, oscillations reappeared under disinhibition after an initial phase of persistent activity. Both spontaneous and disinhibition-induced oscillations were facilitated by riluzole, which enhances fast sodium current inactivation and thus leads to early cessation of firing during strong depolarization (depolarization block). In single cell recordings, episodes of strong depolarization were mostly seen during oscillations induced by disinhibition, but occasionally also during spontaneous oscillations. We conclude that both GABA A-mediated synaptic inhibition and depolarization block contribute to the de-activation of spinal cord networks during oscillation cycles. PMID- 18412629 TI - Interaction between neuropeptide Y (NPY) and brain-derived neurotrophic factor in NPY-mediated neuroprotection against excitotoxicity: a role for microglia. AB - The neuroprotective effect of neuropeptide Y (NPY) receptor activation was investigated in organotypic mouse hippocampal slice cultures exposed to the glutamate receptor agonist alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA). Exposure of 2-week-old slice cultures, derived from 7-day-old C57BL/6 mice, to 8 microm AMPA, for 24 h, induced degeneration of CA1 and CA3 pyramidal cells, as measured by cellular uptake of propidium iodide (PI). A significant neuroprotection, with a reduction of PI uptake in CA1 and CA3 pyramidal cell layers, was observed after incubation with a Y(2) receptor agonist [NPY(13-36), 300 nm]. This effect was sensitive to the presence of the selective Y(2) receptor antagonist (BIIE0246, 1 microm), but was not affected by addition of TrkB-Fc or by a neutralizing antibody against brain-derived neurotrophic factor (BDNF). Moreover, addition of a Y(1) receptor antagonist (BIBP3226, 1 microm) or a NPY-neutralizing antibody helped to disclose a neuroprotective role of endogenous NPY in CA1 region. Cultures exposed to 8 microm AMPA for 24 h, displayed, as measured by an enzyme-linked immunosorbent assay, a significant increase in BDNF. In such cultures there was an up-regulation of neuronal TrkB immunoreactivity, as well as the presence of BDNF-immunoreactive microglial cells at sites of injury. Thus, an increase of AMPA-receptor mediated neurodegeneration, in the mouse hippocampus, was prevented by neuroprotective pathways activated by NPY receptors (Y(1) and Y(2)), which can be affected by BDNF released by microglia and neurons. PMID- 18412630 TI - Neurotoxic and neurotrophic roles of proNGF and the receptor sortilin in the adult and ageing nervous system. AB - The precursor form of the nerve growth factor (proNGF), forms a heterotrimeric complex with the receptors p75 and sortilin; this complex has been implicated in neuron cell death. However, it is not known whether proNGF and the receptors p75 and sortilin contribute to age- and disease-related neurodegeneration. Here we show that proNGF induces cell death in subpopulations of basal forebrain and peripheral sympathetic neurons of old, but not of young, adult rodents. In contrast, proNGF appears to induce neurite outgrowth rather than cell death of young adult sympathetic neurons. We have examined the neurotoxic role of proNGF in old age, and find that proNGF protein is elevated during ageing in the projection areas of some populations of vulnerable central and peripheral neurons; caloric restriction, which has known neuroprotective effects, partially prevents these increases. Sortilin was found to play a significant part in the observed patterns of age-related proNGF-mediated neurotoxicity. In particular, survival of aged neurons was rescued by neurotensin, an alternative sortilin ligand that blocks the sortilin-mediated effects of proNGF. Furthermore, sortilin immunoreactivity increases markedly in ageing rodent basal forebrain and sympathetic neurons; in contrast, p75 levels are either unchanged or reduced. From these data we propose that selective age-related neuronal atrophy and neurodegeneration may be mediated by increased sortilin expression in neurons, together with elevated levels of proNGF expression in some targets. PMID- 18412631 TI - Cerebellar pathology in transgenic mice expressing the pseudorabies virus immediate-early protein IE180. AB - Pseudorabies virus is an alphaherpesvirus causing fatal neurological diseases in animals. Pseudorabies virus carries a gene encoding immediate-early (IE) protein IE180, which controls the transcription of other viral and host cell genes. Previously, we reported that transgenic expression of IE180 in mice causes severe ataxia and cerebellar deformity. Here we identified profound abnormalities in adult IE180 transgenic mice, including malpositioning of Purkinje cells (PCs), granule cells (GCs) and Bergmann glia (BG), impaired dendritogenesis and synaptogenesis in PCs, disoriented BG fibers, absence of molecular layer interneurons, and increased apoptosis of neurons and glia. In accordance with the cellular defects, we found the expression of IE180 in PCs, GCs and astrocytes during cerebellar development. We next examined transgenic mice expressing truncated IE180 mutants: dlN132 lacking the acidic transcriptional active domain, dlC629 lacking the nuclear localization signal and dlC1081 having all known domains but lacking the carboxyl-terminal sequence. Despite similar expression levels of the transgenes, ataxia and cerebellar defects were only manifested in the dlC1081 transgenic mice but their phenotypes were milder compared with the IE180 transgenic mice. In the dlC1081 transgenic mice, cerebellar neurons and glia were normally positioned but cerebellar size was severely reduced due to GC deficits. Interestingly, dlC1081 was mainly expressed in the GCs with low expression in a few BG. Taken together, the present findings clarified a causal relationship between cerebellar pathology and cellular expression of IE180, and further afforded an experimental insight into different symptomatic severity as a consequence of different cellular defects caused by such cytotoxic viral agents. PMID- 18412632 TI - Stress accelerates neural degeneration and exaggerates motor symptoms in a rat model of Parkinson's disease. AB - The causes of most cases of Parkinson's disease (PD) are still poorly understood. Here we show that chronic stress and elevated corticosterone levels exaggerate motor deficits and neurodegenerative events in a Parkinson's disease rat model. Animals were tested in skilled and non-skilled movement while being exposed to daily restraint stress or oral corticosterone treatment. Stress and corticosterone compromised normal motor function and exaggerated motor deficits caused by unilateral 6-hydroxydopamine lesion of the nigrostriatal bundle. Moreover, stress and corticosterone treatments diminished the ability to acquire compensatory strategies in limb use during skilled reaching and skilled walking. In contrast, lesion control animals were able to significantly improve in the ability of skilled limb use during the repeated test sessions. The exaggerated motor impairments in stress-treated animals were related to accelerated loss of midbrain dopamine-producing neurons during the first week postlesion. Correlation analysis revealed a significant connection between loss of tyrosine hydroxylase positive cells and increase in Fluoro-Jade-positive cells only in stress- and corticosterone-treated animals. Furthermore, stress and elevated corticosterone levels caused greater permanent loss of midbrain neurons than found in non treated lesion animals. These findings demonstrate that stress and elevated corticosterone levels can exaggerate nigral neuronal loss and motor symptoms in a rat analogue of PD. It is therefore possible that stress represents a key factor in the pathogenesis of human PD by impeding functional and structural compensation and exaggerating neurodegenerative processes. PMID- 18412633 TI - Stress-induced analgesia in mice: evidence for interaction between endocannabinoids and cholecystokinin. AB - Stress-induced analgesia (SIA) is an important endogenous mechanism in response to stressful stimuli. Opioid peptides, as well as endocannabinoids, are known mediators of SIA. We were interested in whether the endocannabinoid tone and the resulting SIA could be influenced by changing the activity of cholecystokinin (CCK) in mice. This hypothesis is related to recent evidence showing a cellular colocalization of neuropeptide CCK and cannabinoid type 1 (CB(1)) receptors in many central nervous system structures. However, the physiological relevance of this colocalization is unknown. Our aim was to test whether abolishing the endogenous CCK tone might influence the cannabinoid-mediated form of SIA. As expected, the CB(1) antagonist rimonabant prevented the development of analgesia in response to footshock-induced stress in wild-type mice. In contrast, CCK type 2 (CCK(2)) receptor-deficient mice developed SIA regardless of rimonabant treatment. Naloxone, an opioid antagonist, antagonized SIA in both wild-type and CCK(2) receptor-deficient homozygous mice. This finding suggests that CCK, through CCK(2) receptors, modulates the action of endocannabinoids. Gene expression analysis revealed an up-regulation of CCK(2) receptor gene in the lumbar spinal cord and mesolimbic area in wild-type mice in response to stress. In addition, wild-type mice displayed up-regulation of genes implicated in endocannabinoid-mediated neurotransmission (elevation of CB(1) receptor, 2 arachidonoylglycerol and the anandamide-synthesizing enzymes sn-1-diacylglycerol lipase alpha and N-acyl-phosphatidylethanolamine-hydrolysing phospholipase D) in response to stress in the lumbar spinal cord and mesolimbic area. We did not find any of these changes in CCK(2) receptor-deficient homozygous mice. Altogether, behavioural and gene expression studies reflect an involvement of CCK(2) receptors in the development of endocannabinoid-mediated SIA. PMID- 18412634 TI - Repeated seizures lead to altered skilled behaviour and are associated with more highly efficacious excitatory synapses. AB - People with epilepsy have a high incidence of interictal behavioural problems that appear to be related to the location of their seizure focus. This study investigated a novel test of the hypotheses that repeated seizures result in behavioural deficits and altered performance during the interictal state, and that those behaviours are related to the presence of more highly efficacious excitatory synapses. We tested these hypotheses by first repeatedly eliciting seizures with electric current through indwelling electrodes in the corpus callosum at the level of the caudal forelimb area of sensorimotor neocortex in the rat. We then assessed learned skilled behaviours that primarily utilize the forelimbs on tasks that are sensitive to the functional integrity of that structure. We observed both behavioural deficits and altered kinematic performance in rats that experienced repeated neocortical seizures relative to an electrode-implanted control group. From a separate set of rats, tissue was prepared for quantification of thickness and excitatory synaptic subtypes from neocortical layer V. We observed significantly increased numbers of perforated synapses that make their connections directly onto the dendritic shaft at 3 weeks following the last seizure. Altered reaching behaviours are likely due to neural reorganization in the neocortex including more efficacious synapses. PMID- 18412636 TI - Adaptations of glutamatergic synapses in the striatum contribute to recovery from cerebellar damage. AB - Recent findings proposed that the cerebellum and the striatum, key structures in motor control, are more interconnected than commonly believed, and that the cerebellum may influence striatal activity. In the present study, the possible changes of synaptic transmission in the striatum of hemicerebellectomized rats have been investigated. Neurophysiological recordings showed a significant facilitation of glutamate transmission in the contralateral striatum occurring early following hemicerebellectomy. This process of synaptic adaptation appears to be relevant for the compensation of cerebellar deficits. Accordingly, pharmacological blockade of glutamate N-methyl-d-aspartate (NMDA) receptors with MK-801 prevented the rearrangement of excitatory synapses in the striatum and interfered with the recovery from motor disturbances in rats with cerebellar lesions. Hemicerebellectomy also perturbed gamma-aminobutyric acid (GABA) transmission in contralateral but not ipsilateral striatum. The present findings advance the role of striatal excitatory transmission in the compensation of cerebellar deficits, providing support to the notion that adaptations of striatal function exert a role in the recovery of cerebellar symptoms. PMID- 18412635 TI - Treatment of epilepsy: the GABA-transaminase inhibitor, vigabatrin, induces neuronal plasticity in the mouse retina. AB - Vigabatrin was a major drug in the treatment of epilepsy until the discovery that it was associated with an irreversible constriction of the visual field. Nevertheless, the drug is still prescribed for infantile spasms and refractory epilepsy. Disorganization of the photoreceptor nuclear layer and cone photoreceptor damage have been described in albino rats. To investigate the vigabatrin-elicited retinal toxicity further, we examined the retinal tissue of albino mice treated with two vigabatrin doses. The higher dose did not always cause the photoreceptor layer disorganization after 1 month of treatment. However, it triggered a massive synaptic plasticity in retinal areas showing a normal layering of the retina. This plasticity was shown by the withdrawal of rod but not cone photoreceptor terminals from the outer plexiform layers towards their cell bodies. Furthermore, both rod bipolar cells and horizontal cells exhibited dendritic sprouting into the photoreceptor nuclear layer. Withdrawing rod photoreceptors appeared to form ectopic contacts with growing postsynaptic dendrites. Indeed, contacts between rods and bipolar cells, and between bipolar cells and horizontal cells were observed deep inside the outer nuclear layer. This neuronal plasticity is highly suggestive of an impaired glutamate release by photoreceptors because similar observations have been reported in different genetically modified mice with deficient synaptic transmission. Such a synaptic deficit is consistent with the decrease in glutamate concentration induced by vigabatrin. This description of the neuronal plasticity associated with vigabatrin provides new insights into its retinal toxicity in epileptic patients. PMID- 18412637 TI - Enhanced cell death in hippocampus and emergence of cognitive impairments following a localized mini-stroke in hippocampus if preceded by a previous episode of acute stress. AB - This series of experiments represents a test of a theory concerning the etiology of age-related cognitive decline, including Alzheimer's disease (AD). The theory suggests that multiple combinations of cofactors produce variants of these disorders. Two factors that have been linked to the etiology of AD, that are of interest to our laboratories, are stress and vascular strokes. The current experiments tested the cofactors theory by evaluating the neuronal and functional effects of localized subthreshold strokes in the hippocampus of different groups of rats. One group experienced episodes of stress prior to stroke induction while the other did not. The results showed that a low dose of endothelin-1 (ET-1) injected into the hippocampus of groups of rats that had previously experienced stressful episodes showed enhanced hippocampal cell death and neurodegeneration that did not occur in the rats that did not experience stress prior to stroke induction. The results also showed that the stressed rats given subthreshold ET-1 injections into the hippocampus showed hippocampal-based learning and memory deficits that were not present in the non-stressed group given the same injections. This pattern of results suggests that individuals that are under stress are more vulnerable to insults to the hippocampus that have little effect on an individual that is not stressed. This vulnerability might be due to the actions of stress hormones, like the glucocorticoids, that have been previously shown to endanger hippocampal neurons. PMID- 18412641 TI - Improving UK trauma care: the NCEPOD trauma report. PMID- 18412642 TI - The influence of changes in end-tidal carbon dioxide upon the Bispectral Index. AB - Carbon dioxide is known to affect consciousness in animals and humans. We surmised that changes in end-tidal carbon dioxide during anaesthesia might affect the Bispectral Index. Twenty-four patients due to undergo surgery were anaesthetised with fentanyl and a propofol infusion. The Bispectral Index, pulse rate and blood pressure were recorded while end-tidal carbon dioxide levels were changed. The patients acted as their own controls as they were subjected to high, normal and low levels of end-tidal carbon dioxide (3-12 kPa) according to a randomised sequence. There were no changes in the Bispectral Index or haemodynamic variables resulting from manipulation of the end-tidal carbon dioxide. At the level of hypnosis involved in this study, changes in end-tidal carbon dioxide, within the range tested, do not result in changes in the Bispectral Index. PMID- 18412643 TI - The effect of pre-operative anxiety on induction of anaesthesia with propofol. AB - In this prospective study, we investigated the effects of anxiety on the induction dose of propofol and subsequent cardiovascular changes in 197 patients. Pre-operative state and trait anxiety scores were measured using the State Trait Anxiety Inventory. Propofol was administered at 40 mg x kg(-1) x h(-1). Propofol dose was recorded at loss of verbal response and when EEG Bispectral Index decreased to 50. Thereafter, propofol infusion rate was reduced to 8 mg x kg(-1) x h(-1). Cardiovascular data were collected for 15 min after starting induction. Maximum percentage decreases in heart rate and mean arterial pressure, and the point at which the latter occurred, were recorded. On multivariate analysis, anxiety scores did not significantly affect propofol dose or cardiovascular end points, although Bispectral Index at loss of verbal response decreased with increasing trait anxiety (p = 0.02). Anxiety, measured using State Trait Anxiety Inventory, does not appear independently to affect the induction characteristics of propofol. PMID- 18412644 TI - Intra-operative awareness in children and post-traumatic stress disorder. AB - Adults who experience intra-operative awareness can develop disturbing long lasting after-effects, such as daytime anxiety, sleep disturbances, nightmares, flashbacks and, in the worst case, a post-traumatic stress disorder (PTSD). It is unknown whether intra-operative awareness has a similar psychological impact in children. We designed the present study in order to evaluate the incidence of psychological symptoms in children who had either confirmed or possible intra operative awareness. Attempts were made to locate 11 children who had been identified in a previous study, approximately 1 year following their experience. A PTSD questionnaire was administered to the children and their parents in order to detect any long-term or short-term psychological symptoms (the 1-month postoperative data were evaluated retrospectively). Factors believed to be associated with PTSD, such as intra-operative perceptions, the children's temperament and cognitive strategies, and the parents' coping strategies, were also analysed. Seven children were successfully located and interviewed and no short or long-term psychological symptoms were identified. None of them offered negative appraisals of the traumatic event and none had displayed dysfunctional behaviour or cognitive strategies. Thus, none of them had developed a PTSD syndrome. In contrast with what has been reported in adults, these children claimed not to have experienced major pain, terror or helplessness during their surgery. Despite the small sample size, the results of the present study suggest that children suffer less psychological sequelae than adults following intra operative awareness. This may be due to the fact that the children reported less frightening intra-operative sensations as compared with the adults, and had less understanding of the anaesthesia procedure, and this may have influenced their appraisal of their awareness and protected them from the full impact of this potentially traumatic experience. PMID- 18412645 TI - The use of N-terminal pro-B type natriuretic peptide in a pre-operative setting to predict left ventricular systolic dysfunction on echocardiogram. AB - Heart failure is a major risk factor for adverse postoperative events following non-cardiac surgery. The use of transthoracic echocardiogram as a pre-operative investigation to assess cardiac dysfunction has limitations in this setting. The N-Terminal fragment of B-Type natriuretic peptide (NT proBNP) has been used in screening for heart failure. We have investigated the use of NT proBNP as a screening tool for left ventricular systolic dysfunction to reduce the requirement for pre-operative echocardiograms. Ninety-eight pre-operative non cardiac surgical patients scheduled to undergo echocardiography were assessed clinically and with an NT proBNP measurement. Echocardiogram was used to define two groups of patients depending on the presence or absence of abnormal left ventricular function and the NT proBNP level was compared between the groups using non-parametric and receiver-operator-characteristic (ROC) curve analysis. In terms of pre-operative screening, a NT proBNP of <38.2 pmol x l(-1) had a 100% negative predictive value in predicting patients with normal left ventricular systolic function and would have prevented the requirement for echocardiogram in 43% of pre-operative patients. NT proBNP was superior to electrocardiological and clinical criteria for detection of a normal echocardiogram. This may have significant impact in the pre-operative assessment of patients undergoing non cardiac surgery. PMID- 18412646 TI - The relationship between arterial PO2 and mixed venous PO2 in response to changes in positive end-expiratory pressure in ventilated patients. AB - The response of arterial PO(2) (P(a)O(2)) to airway pressure has been used as a measure of recruitment in mechanically ventilated patients. We hypothesised that mixed venous PO(2) (P(mv)O(2)) directly affects P(a)O(2). Sixteen patients with acute lung injury (ALI, lung injury score > or = 1) on volume-controlled mechanical ventilation (F(I)O(2) 0.40) were studied. Positive end-expiratory pressure (PEEP) was increased and decreased. Incremental PEEP increased median values of P(a)O(2), diminished venous admixture (Q(va)/Q(t)) and cardiac index, but maintained arterial PCO(2) and tissue O(2) uptake. These changes were reversed during decremental PEEP. However P(a)O(2) did not increase in 37% of PEEP steps and changes in P(a)O(2) correlated to those in P(mv)O(2) (r(s) = 0.45, p < 0.001). Changes in P(mv)O(2) contributed to changes in Q(va)/Q(t) in determining changes in P(a)O(2) (p < 0.05). P(mv)O(2) may be an independent determinant of P(a)O(2) during mechanical ventilation for ALI, so that dosing PEEP to recruit the lung should not be guided by arterial blood oxygenation alone. Arterial hypoxaemia with increasing PEEP may improve by reducing PEEP (or increasing tissue O(2) delivery), when the fall in P(mv)O(2) is greater than about 0.133 kPa. PMID- 18412647 TI - The effect of lidocaine on remifentanil-induced cough. AB - This study was performed to investigate the incidence of remifentanil-induced cough and evaluate the efficacy of lidocaine on its prevention. Five-hundred patients, aged 18-70 years, were randomly allocated into two groups to receive either lidocaine 0.5 mg x kg(-1) or 0.9% normal saline intravenously 1 min before remifentanil administration at a target effect-site concentration of 4 ng x ml( 1). Any episode of cough was classified as coughing and graded as mild (1-2), moderate (3-4) or severe (5 or more). We found that the overall incidence of cough was significantly higher in the saline group (69 patients; 27.6%) than that in the lidocaine group (38 patients; 15.2%) (p < 0.001). The results of logistic regression indicated that age and smoking were associated with remifentanil induced cough. This study demonstrated that intravenously administered lidocaine 0.5 mg x kg(-1) effectively suppresses remifentanil-induced cough without possible systemic lidocaine toxicity. PMID- 18412648 TI - Determinants of hospital costs associated with traumatic brain injury in England and Wales. AB - Using data from the Trauma Audit Research Network, we investigated the costs of acute care in patients > or = 18 years of age hospitalised for traumatic brain injury between January 2000 and December 2005 in England and Wales. Traumatic brain injury patients were defined and stratified using the Abbreviated Injury Scale. A total of 6484 traumatic brain injury patients were identified; 22.3% had an Abbreviated Injury Scale score of three, 38.0% of four and 39.7% of five. Median age (IQR) was 42 years (28-59) and 76.7% were men. Primary cause of injury was motor vehicle collisions (42.4%) followed by falls (38.0%). In total 23.7% of the patients died before discharge. Hospitalisation costs averaged 15,462 pounds sterling (SD 16,844 pounds sterling). Costs varied significantly by age, Glasgow Coma Score, Injury Severity Score, coexisting injuries of the thorax, spine and lower limb, hospital mortality, availability of neurosurgical services, and specialty of attendants seen in the Accident and Emergency department. PMID- 18412649 TI - A prospective observational study of ICU patient position and frequency of turning. AB - Positioning and turning critically ill patients may be beneficial but there are little data on current practice. We prospectively recorded patient position every hour over two separate days in 40 British intensive care units and analysed 393 sets of observation. Five patients were prone at any time and 3.8% (day 1) and 5% (day 2) were on rotating beds. Patients were on their back for 46.1% of observations, turned left for 28.4% and right for 25.5%, and head up for 97.4%. A turn was defined as a change between on back, turned left or turned right. The average time (SD) between turns was 4.85 (3.3) h. There was no significant association between the average time between turns and age, weight, height, gender, respiratory diagnosis, intubated and ventilated, sedation score, day of week or nurse:patient ratio. There was a significant difference between hospitals in the frequency with which patients were turned. PMID- 18412650 TI - A national survey of safe practice with epidural analgesia in obstetric units. AB - The National Patient Safety Agency (NPSA) identified practice improvements with regard to epidural injections and infusions and released a patient safety alert on 28th March 2007. Prior to this, the Obstetric Anaesthetists' Association had considered the draft document and wished to assess current compliance in UK obstetric units. A postal survey of consultant-led obstetric anaesthetic units in the UK was performed in September 2006 to look at practice prior to the release of the safety alert. The response rate was 89%. Many units are already following the guidance from the NPSA but nearly one in four units have experience of wrong route drug errors related to confusion between systems for intravenous and regional drug administration. PMID- 18412651 TI - Lightwand guided intubation in paediatric patients with a known difficult airway: a report of four cases. AB - The anaesthetic management of children with craniofacial abnormalities often presents unique challenges because soft tissue and bony abnormalities can affect the airway and influence airway management. We report four paediatric patients with predicted difficult airways due to craniofacial abnormalities. They all had a laryngeal view of Cormack-Lehane grade IV and were impossible to intubate using direct laryngoscopy. Fibreoptic intubation was also repeatedly attempted but was not successful. All the tracheal intubations were completed using a lightwand on the first attempt in less than 30 s. We consider that lightwand guided intubation technique may be a useful alternative approach to fibreoptic intubation technique in managing the difficult paediatric airway. PMID- 18412652 TI - Controlled non-heart beating organ donation: neither the whole solution nor a step too far. AB - While many intensive care clinicians in the UK continue to express significant concerns regarding controlled non-heart beating organ donation, others are involved in established programmes that make an increasingly significant contribution to the total number of cadaveric donations each year. The successful introduction of a controlled non-heart beating organ donation programme requires local resolution of any apparent ethicolegal obstacles to the process, with specific attention needing to be given to three areas: the potential conflict of interest between decision making over futility and any subsequent approach regarding organ donation; a belief that it may be unlawful to adjust in any way an end of life care pathway in order to allow donation to take place, and, finally, an uncertainty over how soon after cardiac death organ retrieval can begin. It is proposed that recent changes in legislation provide, through an emphasis on patient autonomy and best interests, a solid ethicolegal foundation for donation after cardiac death. PMID- 18412653 TI - Use of the HemoCue near patient testing device to measure the concentration of haemoglobin in suction fluid at elective caesarean section. AB - We have investigated the suitability of the HemoCue photometer to measure the concentration of haemoglobin in suction fluid obtained at elective caesarean section in 30 women. Laboratory analysis was used as a gold standard against which values generated by the HemoCue were compared. We used the method of Bland and Altman to analyse the data. The bias and the limits of agreement were -0.013 and -0.39 to 0.36 mg x dl(-1) respectively, indicating a good level of agreement. Mean (SD) total blood loss calculated using these data, combined with the weight of the swabs, was consistently greater than clinical estimation: 768 (496) ml versus 506 (249) ml respectively (p < 0.001). We have found that the HemoCue near patient testing device may be used to estimate blood loss accurately in the suction fluid obtained at elective Caesarean section. PMID- 18412654 TI - Techniques for emergency ventilation through a needle cricothyroidotomy. AB - We examined the pressures produced by a construction intended for emergency ventilation through a needle cricothyroidotomy. This construction consisted of a standard hospital wall oxygen supply, flowmeter, oxygen tubing and a three-way tap. We measured the flow achieved through a transtracheal catheter and compared the construction to a Manujet jet ventilator and to a Sanders injector. The construction performed similarly to the Manujet set at low pressures (0-100 kPa). To achieve similar pressures and flow to the Manujet set at pressures higher than 100 kPa required opening of the flowmeter beyond its calibrated range. The flow through the transtracheal catheter was almost three times higher when the flowmeters were fully opened than when they were opened to the 15 l x min(-1) mark (44.5 vs 15.8 l x min(-1), respectively; p < 0.0001). When the flowmeters were fully opened the pressure measured before the catheter was over four times higher than when they were only opened to the 15 l x min(-1) mark (285.3 vs 66.4 kPa, respectively; p < 0.0001). This system of ventilation is inferior to a Manujet in terms of robustness and calibration throughout its range of pressures and flows, but seems appropriate for emergency use in the absence of a purpose made jet ventilator. PMID- 18412655 TI - Malignant hyperthermia presenting during laparoscopic adrenalectomy. AB - A 44-year-old man presented for elective laparoscopic adrenalectomy. During the procedure his end-tidal carbon dioxide readings rose steadily. We assumed that this was due to a prolonged carbon dioxide pneumoperitoneum until he developed ST segment depression on his electrocardiogram and a rapid rise in temperature. A diagnosis of malignant hyperthermia was made in view of the rising temperature and carbon dioxide. He responded to cooling and intravenous dantrolene. He was later confirmed to be malignant hyperthermia-susceptible on in vitro contracture testing of a muscle biopsy. The diagnosis was delayed as the early signs of malignant hyperthermia are the same as the expected physiological changes in laparoscopic surgery. As laparoscopic surgery continues to expand we advocate vigilance to ensure early identification of this rare but potentially devastating condition. PMID- 18412656 TI - Caesarean section in a complicated case of central core disease. AB - We describe the anaesthetic management of a 21-year-old lady with central core disease for elective Caesarean section. Central core disease is characterised by muscle weakness, skeletal deformities and susceptibility to malignant hyperthermia. Total intravenous anaesthesia was used because of the combination of potential malignant hyperthermia, severe kyphoscoliosis and extensive spinal scarring. The authors believe there is no previous report of propofol and remifentanil being used in these circumstances. A short review of central core disease and its anaesthetic implications is provided. PMID- 18412657 TI - Postoperative muscle weakness in a patient recently treated with infliximab. AB - We present a case of profound postoperative muscle weakness in a patient who had been treated with infliximab, and whose weakness had not manifested pre operatively. We believe this to be the first case report of infliximab-related muscle weakness manifesting immediately after anaesthesia. PMID- 18412659 TI - Interpleural block and respiratory side effects. PMID- 18412660 TI - Interpleural block and safe high quality analgesia after thoracotomy. PMID- 18412661 TI - Epidural analgesia: first do no harm. PMID- 18412662 TI - Interpleural blocks and clotting abnormalities: a case report. PMID- 18412664 TI - Preventing pacemaker problems with nerve stimulators. PMID- 18412667 TI - Bilateral greater occipital nerve block for post-dural puncture headache. PMID- 18412668 TI - Hypertonic saline after traumatic brain injury in UK neuro-critical care practice. PMID- 18412669 TI - Tissue adhesive as an alternative to sutures for securing central venous catheters. PMID- 18412670 TI - Unilateral sensorineural hearing loss after general anaesthesia. PMID- 18412671 TI - Newton valve: a word of caution. PMID- 18412672 TI - How do you fix reinforced tracheal tubes? PMID- 18412676 TI - Changes in ANZJPH Instructions for Authors. PMID- 18412677 TI - Ten citation classics from the Australian and New Zealand Journal of Public Health. PMID- 18412678 TI - Window of opportunity--positioning food and nutrition policy within a sustainability agenda. AB - Public health professionals have an opportunity to refocus national attention on food and nutrition policy, within a sustainability agenda. A broadly based national Food and Nutrition Policy was developed in 1992. However, its implementation has been selective and primarily based within the health sector. Other major policy areas, for example; industry, agriculture and trade, have dominated Australian nutrition and health policy. A broad, whole-of-government commitment to a comprehensive food and nutrition policy that engages with the community is required to achieve outcomes in terms of public health, a sustainable environment and viable food production for future generations. PMID- 18412679 TI - The social determinants of being an Indigenous non-smoker. AB - OBJECTIVE: To examine the association between various social factors and being a non-smoker in a national survey of Aboriginal and Torres Strait Islander people aged 15 and over. METHODS: We analysed data from the 2002 National Aboriginal and Torres Strait Islander Social Survey (n=9,400) using logistic regression. RESULTS: About a half (51.2%) of the Aboriginal and Torres Strait Islander population aged 15 years and over smoked, 33.4% had never smoked, and 15.4% were ex-smokers. Higher socio-economic position (as measured by each of nine variables) was strongly associated with being a non-smoker rather than a smoker, after controlling for age and gender. There was a clear income gradient: increasing household income was associated with increasing likelihood of being a non-smoker. Indigenous people who had been arrested in the last five years were 4.5 times less likely to be non-smokers, adjusted for age and gender. Indigenous people who had been removed from their natural family were half as likely to be a non-smoker. CONCLUSIONS: Different groups within the Indigenous population have quite different smoking behaviours, although the prevalence of smoking is very high in all groups. The poorest and most socially disadvantaged are the least likely to be non-smokers. IMPLICATIONS: Indigenous tobacco control programs need to consider additional targeting of more disadvantaged groups. Tobacco control programs should work with broader campaigns to ameliorate social disadvantage among Indigenous peoples. PMID- 18412680 TI - Did it fall or was it pushed? The contribution of trends in established risk factors to the decline in premature coronary heart disease mortality in New Zealand. AB - OBJECTIVE: To estimate the contribution of trends in three risk factors--systolic blood pressure (SBP), total blood cholesterol (TBC) and cigarette smoking--to the decline in premature coronary heart disease (CHD) mortality in New Zealand from 1980-2004. METHOD: Risk factor prevalence data by 10-year age group (35-64 years) and sex was sourced from six national or Auckland regional health surveys and three population censuses (the latter only for smoking). The data were smoothed using two-point moving averages, then further smoothed by fitting quadratic regression equations (SBP and TBC) or splines (smoking). Risk factor/CHD mortality hazard ratios estimated by expert working groups for the World Health Organization Global Burden of Disease Study 2001 were used to translate average annual changes in risk factor prevalences to the corresponding percentage changes in premature CHD mortality. The expected trends in CHD mortality were then compared with the observed trend to estimate the contribution of each risk factor to the decline. FINDINGS: Approximately 80% (73% for males, 87% for females) of the decline in premature CHD mortality from 1980 to 2004 is estimated to have resulted from the joint trends in population SBP and TBC distributions and smoking prevalence. Overall, approximately 42%, 36% and 22% of the joint risk factor effect was contributed by trends in SBP, TBC and smoking respectively. CONCLUSION: Our estimate for the joint risk factor contribution to the CHD mortality decline of 80% exceeds those of two earlier New Zealand studies, but agrees closely with a similar Australian study. This provides an indicator of the scope that still remains for further reduction in CHD mortality through primary and secondary prevention. PMID- 18412681 TI - Deaths due to suicide: the effects of certification and coding practices in Australia. AB - OBJECTIVE: To highlight issues relating to suicide coding that have an impact on the final reported mortality data by the Australian Bureau of Statistics. METHOD: This paper presents an outline of the way in which the official Australian suicide data is captured and coded and highlights issues relating to the classification used, coronial processes, documentation requirements and data sources that have an impact on the final reported data, especially deaths associated with coronial investigations. RESULTS: Issues related to the coding of Australian suicide data are: *Disparity between jurisdictions due to differences in documentation about accidental or undetermined causes of death. *Lack of standardisation in the way that coronial deaths are reported across Australia. *Lack of a standard form for police reports. *Administrative processes that cause delays in reporting the results of coronial investigations. *Reluctance on the part of some coroners to report deaths as suicides. CONCLUSIONS: Researchers and policy makers need to be aware of the constraints under which suicide can be reported as such in the official data before interpreting time trends. PMID- 18412682 TI - Residential mobility in a cohort of primiparous women during pregnancy and post partum. AB - OBJECTIVE: To quantify the residential mobility rate in a population of pregnant women expecting their first baby. METHOD: We verified residential mobility in a cohort of 585 primiparous Australian women who were enrolled in late pregnancy and had participated in a randomised controlled trial and followed-up to at least 16 weeks post-partum. RESULTS: We found a residential mobility rate of 19%. Movers and non-movers differed by socio-demographic factors, with movers more likely be younger, relative risk (RR)=2.14 (95% confidence interval (CI) 1.41 3.13), and not living with a partner RR=2.46 (95% CI 1.60-3.77). CONCLUSION: Most prospective epidemiological studies can expect some attrition in the study population. The family formation period is acknowledged as a highly mobile time and this mobility may contribute to loss to follow-up. IMPLICATIONS: Researchers planning prospective studies in pregnant populations should consider the impact of residential mobility, especially differential mobility, and implement strategies to reduce attrition and optimise response rates. PMID- 18412683 TI - Perceived weight versus Body Mass Index among urban Aboriginal Australians: do perceptions and measurements match? AB - OBJECTIVE: To examine the relationship between perceived body weight and measured Body Mass Index (BMI) among urban Aboriginal Australian adults. METHODS: We compared responses to a question on perceived weight with BMI based on measured health and weight among 248 Aboriginal volunteers aged>or=15 years who took part in a larger health study in the Darwin area between September 2003 and March 2004. Logistic regression was used to examine associations between socio economic, demographic and cultural factors and under-assessment of weight. RESULTS: Being male and having diabetes were significantly associated with under assessment of weight. Despite under-assessment being common, most participants with a BMI>or=25--and almost all (>90%) those with BMI>or=25 plus high waist circumference--described themselves as overweight. CONCLUSIONS: Study participants with BMI>or=25 were generally able to classify themselves appropriately as overweight. IMPLICATIONS: Lack of awareness of weight is unlikely to represent a major barrier to engaging Aboriginal people. However, other barriers exist, and both individual-level and environmental/structural approaches are required to reduce the burden of obesity among Aboriginal Australians. PMID- 18412684 TI - Chronic obstructive pulmonary disease (COPD) is a major personal and public health burden in Australia. AB - OBJECTIVE: Chronic obstructive pulmonary disease (COPD) is a serious disease that is increasing in prevalence, yet is under-diagnosed and under-recognised in Australia. The goal is to bring this to the attention of public health and offer recommendations in order to reduce the present and future burden from COPD. APPROACH: The increase in prevalence, particularly in women, and the impact upon personal and public health demonstrate that COPD is a major public health issue. A proactive approach for public health is suggested, including strategies to increase smoking cessation among at-risk groups, reduce delayed diagnosis, extend disease management strategies, and establish an Australian research base. CONCLUSION: The reduction of the burden from COPD is contingent upon a proactive approach by public health that includes approaching it as a research priority and the provision of adequately resourced prevention and management strategies. PMID- 18412685 TI - Migrants and tuberculosis: analysing epidemiological data with ethnography. AB - OBJECTIVE: Media portrayals of tuberculosis (TB) in New Zealand are of immigrants who enter the country with active disease and pose a threat to inhabitants, which fosters a popular perception that border control is the best and only response to disease control. This paper reviews both New Zealand and international data on TB rates, causes and transmission among migrant populations to elucidate the precise nature of the link between immigration and TB rates. METHODS: Recent information from scholarly journals on immigration and TB was reviewed. Surveillance data from New Zealand and comparable information from other low-incidence countries were reviewed. CONCLUSIONS AND IMPLICATIONS: The importation of active TB is only a minor part of the total TB burden. While effective border control is essential, equally, if not more important, are the circumstances that promote the reactivation of latent TB infection in migrant communities, including migrants' experiences in transit and after arrival, structural conditions, and personal characteristics. For sound prevention strategies, attention needs to be paid to the existence of transnational communities and the conditions for migrants, rather than placing a singular focus on place of birth. PMID- 18412686 TI - Physical activity, Body Mass Index and health care costs in mid-age Australian women. AB - OBJECTIVE: This study examined the relationships between combined categories of physical activity (PA) and Body Mass Index (BMI) with health care costs in women and assessed the potential cost savings of improving PA and BMI in sedentary mid age women. METHODS: Cross-sectional analysis of 2001 survey data linked to health service use data for the same year from 7,004 mid-age women (50-55 years) participating in the Australian Longitudinal Study on Women's Health. RESULTS: The mean (median; interquartile range) annual cost of Medicare-subsidised services was $542 (355; 156-693) per woman. Costs were 17% higher in obese than in healthy-weight women and 26% higher in sedentary than in moderately active women. For sedentary obese women, mean costs were 43% higher than in healthy weight, moderately active women. After adjustment for potential confounders, the relative risk of 'high' claims (>or=15 claims per year) for overweight women who reported 'moderate' or 'high' PA were lower than for women with healthy BMI who reported no PA. CONCLUSIONS AND IMPLICATIONS: Lower PA and higher BMI are both associated with higher health care costs, but costs are lower for overweight active women than for healthy-weight sedentary women. At the population level these data suggest that there would be significant cost savings if all sedentary mid-age women could achieve at least 'low' levels of PA (60-150 minutes a week). PMID- 18412687 TI - Estimating the costs of drug-related hospital separations in Australia. AB - OBJECTIVE: To estimate the total hospital costs of drug-related separations in Australia from 1999/2000 to 2004/05, and separate costs for the following illicit drug classes: opioids, amphetamine, cannabis and cocaine. METHODS: Australian hospital separations between 1999/2000 to 2004/05 from the National Hospital Morbidity Dataset (NHMD) with a principal diagnosis of opioids, amphetamine, cannabis or cocaine were included, as were indirect estimates of additional 'drug caused' separations using aetiological fractions. The costs were estimated using the year-specific case weights and costs for each respective Diagnostic Related Group (DRG). RESULTS: Total constant costs decreased from $50.8 million in 1999/2000 to $43.8 million in 2002/03 then increased to $46.7 million in 2004/05. The initial decrease was driven by a decline in numbers of opioid-related separations (with costs decreasing by $11.5 million) between 1999/2000 and 2001/02. Decreases were evident in separations within the opioid use, dependence and poisoning DRGs. Increases in costs were observed between 1999/00 and 2004/05 for amphetamine (an increase of $2.4 million), cannabis ($1.8 million) and cocaine ($330,000) related separations. Several uncommon but very expensive drug related separations constituted 12.7% of the total drug-related separations. CONCLUSIONS AND IMPLICATIONS: Overall, the costs of drug-related hospital separations have decreased by $4.1 million between 1999 and 2005, which is primarily attributable to fewer opioid-related separations. Small reductions in the number of costly separations through harm reduction strategies have the potential to significantly reduce drug-related hospital costs. PMID- 18412688 TI - NZ Government's trend analysis of hospitalised self-harm is misleading. AB - OBJECTIVE: The aim of this paper is to demonstrate that the trends published in the New Zealand (NZ) Government's 2006 Suicide Trends document for hospitalised self-harm are misleading. METHODS: Analysis of incident self-harm events resulting in hospitalisation and reference to published material on injury outcome indicators for the NZ Injury Prevention Strategy (NZIPS). RESULTS: The significant increase in rates of self-harm hospitalisation presented in Suicide Trends from 1989 to a large extent reflect changes in recording practice rather than any change in self-harm in the community. Indicators with significantly fewer threats to validity suggest there has been little, if any, increase in the incidence of self-harm. The authors of Suicide Trends did not adequately specify how they defined a case and, moreover, their methods were not consistent with those used for the NZIPS indicators. CONCLUSIONS AND IMPLICATIONS: The methodological challenges to producing valid indicators for the purposes of measuring trends in important non-fatal injury are substantial. Unless we accept that the usual methods of measuring trends in non-fatal injury are misleading and commit to taking up the challenge to produce and use better indicators, we will continue to run the risk of misleading ourselves and the public. PMID- 18412689 TI - Lower limb injuries in New Zealand Defence Force personnel: descriptive epidemiology. AB - OBJECTIVE: To describe the epidemiology of lower limb injuries in the New Zealand Defence Force (NZDF). METHOD: Data from all NZDF lower limb injury claims from an 11-month period were examined for type, site, and circumstances of injury. Both injury codes and narratives were analysed, allowing each injury event to be classified according to mechanism of injury, object involvement, and activity at the time of injury, as well as type and site. RESULTS: The commonest lower limb musculoskeletal injuries were ankle sprains or strains (35%) and knee sprains or strains (16%). Most commonly, injuries were due to acute over-exertion (37%), involved no other person (50%), and occurred while running (28%) or playing team sports (25%). The injury rate for recruits was more than five times that of trained personnel. CONCLUSIONS: Potential interventions should target ankle sprains primarily, but also knee sprains and fractures. Fractures, while accounting for only 6% of lower limb injuries, should be a priority because of their high medical and time-lost costs. Interventions must also take into account the high incidence of injuries involving individuals alone and sustained during recruit training. The study also demonstrated that analysis of military injury narratives provides valuable extra information on injury causation and the circumstances of injury, and allows more accurate characterisation of the injury process. IMPLICATIONS: This study will provide the basis for development of an injury prevention strategy for lower limb training injuries in the NZDF. PMID- 18412690 TI - Internet access for delivery of health information to South Australians older than 50. AB - OBJECTIVE: An exploratory study to determine the proportion of people aged 50-76 years who have Internet access, the location of this access, predictive characteristics of such access, and preparedness to receive unsolicited health information through this medium. METHODS: A random sample of 1,000 South Australians was identified from the Australian Electoral Commission roll and contacted by telephone in 2006. They were invited to answer questions about their access to the Internet and attitude towards receiving unsolicited health information via the Internet. RESULTS: Of those contacted, 41% provided information. Of this group, 59% indicated that they had Internet access, mostly at home. Men were more likely than women to have home access (p<0.05). Higher educational achievement, higher economic status and younger age were all significantly associated with access. Of those who had access, 65% would be willing to receive health information through the Internet. CONCLUSIONS: More than half the population older than 50 years has access to the Internet at some location, and there is a general acceptance of the possibility of receiving health-promoting material via this medium. However, delivery via the Internet alone would disadvantage those who are less educated, less financially well off and older. IMPLICATIONS: It is likely that a generational change in the entire nature of public communication will influence the efficiency and effectiveness of preventive health promotion delivery via the Internet. In order to facilitate improved health outcomes, issues of disparity of access must be addressed. PMID- 18412691 TI - Innovation and effectiveness: changing the scope of school nurses in New Zealand secondary schools. AB - OBJECTIVE: To describe the changing role of school nurses in eight New Zealand (NZ) secondary schools from low socio-economic areas with high Pacific Island and Maori rolls. METHODS: An evaluation of a pilot addressing under-achievement in low-decile schools in Auckland, NZ (2002-05). Annual semi-structured school nurse interviews and analysis of routinely collected school health service data were undertaken. RESULTS: Two patterns of school nurse operation were identified: an embracing pattern, where nurses embraced the concept of providing school-based health services; and a Band-Aid pattern, where only the basics for student health care were provided by school nurses. CONCLUSIONS AND IMPLICATIONS: School nurses with an embracing pattern of practice provided more effective school-based health services. School health services are better served by nurses with structured postgraduate education that fosters the development of a nurse-practitioner role. Co-ordination of school nurses either at a regional or national level is required. PMID- 18412692 TI - Is concern about waterpipe tobacco smoking warranted? PMID- 18412693 TI - Response to Calver et al. on the WA regulatory scheme for stimulants. PMID- 18412694 TI - Do Australians perceive their weight status differentially and accurately? Implications for health promotion. PMID- 18412695 TI - The effectiveness of the provision of free nicotine replacement therapy on quit rates among health staff. PMID- 18412709 TI - Infection control measures to limit the spread of Clostridium difficile. Introduction. PMID- 18412710 TI - Infection control measures to limit the spread of Clostridium difficile. AB - Clostridium difficile-associated diarrhoea (CDAD) presents mainly as a nosocomial infection, usually after antimicrobial therapy. Many outbreaks have been attributed to C. difficile, some due to a new hyper-virulent strain that may cause more severe disease and a worse patient outcome. As a result of CDAD, large numbers of C. difficile spores may be excreted by affected patients. Spores then survive for months in the environment; they cannot be destroyed by standard alcohol-based hand disinfection, and persist despite usual environmental cleaning agents. All these factors increase the risk of C. difficile transmission. Once CDAD is diagnosed in a patient, immediate implementation of appropriate infection control measures is mandatory in order to prevent further spread within the hospital. The quality and quantity of antibiotic prescribing should be reviewed to minimise the selective pressure for CDAD. This article provides a review of the literature that can be used for evidence-based guidelines to limit the spread of C. difficile. These include early diagnosis of CDAD, surveillance of CDAD cases, education of staff, appropriate use of isolation precautions, hand hygiene, protective clothing, environmental cleaning and cleaning of medical equipment, good antibiotic stewardship, and specific measures during outbreaks. Existing local protocols and practices for the control of C. difficile should be carefully reviewed and modified if necessary. PMID- 18412712 TI - The multidisciplinary management of rectal cancer. PMID- 18412714 TI - Evaluation of a radiographer-provided barium enema service. AB - AIM: Radiographers performed and reported 5516 double contrast barium enemas (RDCBEs) over 4 years to October 2001. This study was undertaken to assess the accuracy of RDCBE and the sensitivity for diagnosing colorectal cancer (CRC). METHOD: A total of 224 consecutive outpatient RDCBEs were reported; normal (C1), diverticulosis (C2), diverticulosis with filling defect (C3), diverticulosis & other pathology (C4) and abnormal (C5). RDCBEs were then reported by a radiologist (AL, NS) and the two reports compared. Of 450 CRCs, 153 had undergone DCBE; 152 RDCBEs. Reports were analysed to establish concurrence between radiographer and radiologist and final CRC diagnosis. RESULTS: By category: C1 - 37%, C2 - 31%, C3 - 21%, C4 - 11%, C5 - 0%. C4s included polyps (50%), cancer (12.5%), disrupted anastomosis (8%) and colitis (4%). There was no discrepancy between RDCBE and radiologist reports. Radiology and CRC diagnosis agreed in 145 of 152 DCBEs. There were three exclusions: DCBEs occurred outside the study period (2), one only with raised possibility of malignancy. Of eight remaining RDCBEs, seven were false negatives and one false positive. Sensitivity score for RDCBE was 94.5%. Double reporting by a radiologist did not improve sensitivity. CONCLUSION: RDCBEs are as accurate as those performed by radiologists and have a very high sensitivity for CRC. In a time of ever increasing demands for complex staging investigations for CRC and interventional radiology, the ACPGBI needs to reconsider its guidelines on radiographers not only performing but also reporting DCBE. PMID- 18412715 TI - Multidisciplinary team meetings for pelvic floor disorders. PMID- 18412718 TI - Scleroderma: considerations for dental hygienists. AB - Scleroderma, the general name of a group of progressive diseases affecting the connective tissues is the most deadly of the varying connective tissue disorders. Characterized by abnormal thickening of the skin, this collagen-vascular disease is associated with immune dysfunction. Hallmark signs of scleroderma include fibrosis, vascular instability and initial inflammation resulting from excessive collagen deposition. Oral facial involvement is considerable, necessitating adaptations in patient oral self-care and influencing oral hygiene. Appropriate dental hygiene management of patients with this autoimmune disorder requires an understanding of clinical characteristics, the recognition of oral facial involvement, treatment considerations and pharmacological interventions. With this information, dental hygienists will be better prepared to provide compassionate, safe and effective dental hygiene management and care to patients with scleroderma. PMID- 18412719 TI - Post-treatment effects of subantimicrobial dose doxycycline on clinical parameters and gingival crevicular fluid transforming growth factor-beta1 in severe, generalized chronic periodontitis. AB - OBJECTIVE: Present study aimed to evaluate the effect of 3-month adjunctive subantimicrobial dose doxycycline (SDD) on clinical parameters and gingival crevicular fluid (GCF) transforming growth factor-beta1 (TGF-beta(1)) levels in chronic periodontitis patients over 12 months. METHODS: Thirty-five patients with severe, generalized periodontitis participated in the present randomized, placebo controlled study. Patients received scaling and root planing (SRP) plus 3 months adjunctive SDD or placebo. Clinical measurements and GCF sampling were performed at baseline, 3, 6, 9 and 12 months. Eleven periodontally healthy subjects served as controls for GCF TGF-beta(1) analysis. RESULTS: Clinical parameters of both SDD and placebo groups significantly improved during the study (P < 0.0125). SDD group exhibited significantly higher PD reduction at deep sites (baseline PD > or =7 mm) compared with placebo group at 6 months (P < 0.05). In SDD group significantly higher percentage of deep pockets resolved (PD reduction > or =3 mm from baseline) when compared with placebo group at 6 and 9 months (73.4% versus 49.7%; 79.9% versus 50.6%, respectively, P < 0.05). PD reduction > or =4 mm for deep pockets from baseline was also greater in SDD group than placebo at 6 months (53.4% versus 36.3%, P < 0.05). GCF TGF-beta(1) levels of SDD group was significantly higher than baseline (P < 0.0125) and placebo group (P < 0.017) at 3 months. CONCLUSIONS: These results ensure further data for beneficial effects of adjunctive SDD therapy in the management of severe chronic periodontitis. PMID- 18412720 TI - Oral health-related quality of life in children: part I. How well do children know themselves? A systematic review. AB - OBJECTIVE: Paediatric oral disorders are likely to have a negative effect on the quality of life. Until recently, children's oral health-related quality of life (OHRQoL) was measured using parents as informants. Instruments have now been developed, which have demonstrated that with appropriate questionnaire techniques, valid and reliable information can be obtained from children. The aim of this study was to make a systematic review of the existing literature about child perceptions of OHRQoL and their validation. METHODS: A computerized search was conducted using Medline, ISI, Lilacs and Scielo for children's perception of OHRQoL. The inclusion criteria were: the articles should contain well-validated instruments and provide child perceptions of OHRQoL. RESULTS: From 89 records found, 13 fulfilled the criteria. All studies included in the critical appraisal of the project suggested good construct validity of overall child perceptions of OHRQoL. However, children's understanding of oral health and well-being are also affected by variables (age, age-related experiences, gender, race, education, culture, experiences related to oral conditions, opportunities for treatment, childhood period of changes, back-translating questionnaire, children self perceived treatment need). CONCLUSIONS: The structure of children's self-concept and health cognition is age-dependent as a result of their continuous cognitive, emotional, social and language development. By using appropriate questionnaire techniques, valid and reliable information can be obtained from children concerning their OHRQoL. PMID- 18412721 TI - Oral health-related quality of life in children: part II. Effects of clinical oral health status. A systematic review. AB - OBJECTIVE: Children are affected by numerous oral and orofacial disorders, which have the potential to compromise functioning, well-being and the quality of life (QoL). The purpose of this paper was to review the literature about children's clinical oral health status and health-related quality of life (HRQoL) and to assess the respective association. MATERIALS AND METHODS: The authors searched Medline, ISI, Lilacs and Scielo for articles from 1985 to 2007. The inclusion criteria were randomized, cross-sectional, longitudinal or retrospective studies that used well-validated oral health-related QoL instruments, children self applied questionnaires and quantitative measurements of clinical oral health status. RESULTS: Of the 402 articles that were critically assessed, 12 studies were included in the critical appraisal of the project. CONCLUSIONS: There is a relationship between clinical oral health status and HRQoL in children. In the studies that suggested weak relationships between children's oral conditions and HRQoL, the explanations were low disease levels in the sample, the conditions under investigation may have caused immeasurably low levels of impact or the impacts were mediated by inter- and intravariables according to culture and education. Moreover, relationships between biological or clinical variables and HRQoL outcomes are not direct, but mediated by a variety or personal, social and environmental variables, as well as by the child development, which have influence on the comprehension about the relationship among health, illness and QoL. So, longitudinal studies are necessary to determine validity, responsiveness and minimal clinically important difference. PMID- 18412722 TI - Oral health-related quality of life in children: part III. Is there agreement between parents in rating their children's oral health-related quality of life? A systematic review. AB - OBJECTIVE: One issue that receives a great deal of attention is the comparison between measurement of children's oral health-related quality of life (OHRQoL) reports with those of their parents. However, the extent to which parents understand the effects of ill-health on their children's lives remains unanswered. The purpose of this systematic review was to identify the literature on the nature, extent and the pattern of agreement/disagreement between parent and child reports about child OHRQoL and assess the association between them. MATERIALS AND METHODS: The literature was searched using MEDLINE, ISI, Lilacs and Scielo, from January 1985 to March 2007. The selected studies used well-validated instruments and provided children's and parent's perceptions of child OHRQoL. RESULTS: A total of 87 articles were retrieved and five were selected for the review, which showed that children and parents do not necessarily share similar views about child OHRQoL. Some parents may have limited knowledge about their children's OHRQoL, particularly the impact on social and emotional well-being. CONCLUSIONS: Valid and reliable information can be obtained from parents and children using appropriate questionnaire techniques. Although the parents' reports may be incomplete due to lack of knowledge about certain experiences, they still provide useful information. PMID- 18412723 TI - Self-reported oral health perceptions of Somali adults in Minnesota: a pilot study. AB - OBJECTIVE: To assess self-reported oral health perceptions and associated factors in an adult Somali population living in Minnesota, USA. METHODS: We analysed data from a cross-sectional study of Somali adults aged 18 to 65+ years attending a dental school clinic for care. A comprehensive oral examination was performed by the dental school outreach team on all patients who attended a 2-week designated Somali dental clinic. Adults who consented were given an oral health questionnaire to collect information on sociodemographics, marital status, language preference and self-rated oral and general health. We performed summary statistics and differences between proportions using Fisher's exact test and a comparison of means using one-way anova or a two-sample t-test. RESULTS: The sample consisted of 53 adults, 75% of whom were females. About 49% of subjects reported poor/fair oral health and 38% reported poor/fair general health. Seventy four percent rated their access to dental care as poor/fair and 83% reported that they did not have a regular source of dental care. Self-rated oral health was significantly associated with marital status (P < 0.05) and self-rated general health (P < 0.01) using Fisher's exact test. CONCLUSION: A substantial proportion of Somali adults rated their oral health and access to dental care as poor/fair. These findings suggest that this population would benefit from improved access to oral health care and culturally appropriate oral health education and promotion programs. PMID- 18412724 TI - Study of oral changes in patients with eating disorders. AB - BACKGROUND: The prevalence of eating disorders is very high in industrialized countries, especially in young women. The principal orodental manifestations are erosion of the dental enamel, caries and dental sensitivity. OBJECTIVES: Our aim was to study oral complications in female with eating disorders. METHODS: We study oral complications (dental, mucosal and salivary) in 17 female patients from the Hospitalized Patients Eating Disorders Unit. RESULTS: We found alterations in salivary flow and pH, as well as salivary gland enlargement. CONCLUSIONS: A significant alteration in oral tissue occurs; this has an adverse impact on oral health, producing an accumulation of local irritants which favour the appearance of oral diseases. Preventative measures are therefore recommended for this group of patients. PMID- 18412725 TI - Effects of surgical removal of mandibular third molar on the periodontium of the second molar. AB - OBJECTIVE: The effects on periodontal tissues of adjacent second molars after semi-impacted mandibular third molar surgery were evaluated. The influence of flap design was studied. METHODS: Twenty volunteers randomly underwent the three cornered flap technique (group A) or the distal wedge flap technique (group B). The periodontal probing depth was measured by using a 'Williams'-type probe just prior to surgery and three months post-operatively. Six sites, mesio-buccal, buccal, disto-buccal, disto-lingual, lingual and mesio-lingual, around the second molar were selected for measurement. Kruskal-Wallis test and Dunn test (post hoc) were used. Significance level was set at 5%. RESULTS: There were no complications (oedema, alveolitis, etc.) in any of the patients of the study. The results showed that both methods caused shallow pocket depth (P > 0.05) and there were no statistically significant differences between the flap techniques (P > 0.05). Flap design was not an important factor affecting the periodontal status of the second molar. CONCLUSION: The decision to use any of the various flap designs for access to mandibular third molars should be based on operator preference rather than on the assumption that periodontal health of the adjacent second molar will be improved. PMID- 18412726 TI - Effects of fixed orthodontic appliances on subgingival microflora. AB - OBJECTIVES: Fixed orthodontic appliances cause plaque accumulation around bands and brackets. Since the microbiological composition of dental plaque is closely connected to periodontal tissue health, the aim of this study was to determine the effects of fixed orthodontic appliances on subgingival microflora and periodontal status. METHODS: This prospective study was carried out on 32 adolescents scheduled for fixed orthodontic treatment. Subgingival dental plaque samples and periodontal records (pocket probing depth and clinical attachment level) were obtained in four recording times: before bonding of fixed appliances (T0), 1 (T1), 3 (T2) and 6 (T3) months after the beginning of orthodontic therapy, in order to detect the changes in periodontopathic anaerobe microbial flora and its effects on periodontal status. RESULTS: The values of pocket probing depth, total number of microorganisms and number of patients with positive findings of Prevotella intermedia and other periodontopathic anaerobes increased from T0 to the maximum obtained in T2 recording time. Both clinical and microbiological values decreased 6 months after the beginning of orthodontic therapy. CONCLUSIONS: The therapy with fixed appliances may transitionally increase the growth of periodontopathogenic bacteria and consequently result in gingival inflammatory response but without destructive effect on deep periodontal tissues. PMID- 18412727 TI - Gingival inflammation assessment by image analysis: measurement and validation. AB - AIM: Gingival inflammation may be caused by injury or plaque-related diseases and reduction in inflammation can be a useful indicator of gingival recovery. There has been little research on development of non-index methods to measure gingival condition. The aims of the study were to investigate the reliability of the measurement of changes in gingival redness and swelling, using image analysis, and to compare this approach with an established method for assessing gingival overgrowth [J Clin Periodontol 28 (2001) 81]. METHOD: Twenty volunteers with gingival inflammation were recruited and digital images were taken. Duplicate measurements were made on the first visit by two examiners. At a subsequent visit following periodontal treatment, second images were taken. Gingival changes were determined by assessing redness and tooth surface area visible between the level of the inter-proximal papillae and the gingival margin. Tooth area measurements were compared with the established gingival overgrowth method. RESULTS: The method showed excellent reliability for both intra- and inter-examiner measurements of 0.968-0.998 and 0.769-0.947, respectively, according to the classification by Donner and Eliasziw of the Fleiss coefficient of reliability (repeat measures taken during the patients' first attendance). High correlation was found for gingival encroachment when compared with the established gingival overgrowth method. CONCLUSION: This technique proved a reliable method for investigating changes in gingival redness. High correlation was found for gingival encroachment when compared with an established method. PMID- 18412728 TI - The effectiveness of a novel optical probe in subgingival calculus detection. AB - OBJECTIVES: The aim of the present study was to evaluate the effectiveness of a novel optical calculus detection system under in vivo conditions. METHODS: One hundred and seventy-six tooth surfaces from 44 adult teeth that were indicated for extraction were selected for the present study. The patients were randomly assigned to one of the two experimental groups. In group A (n = 96), clinical presence or absence of subgingival calculus deposits was determined using the light-emitting diode-based optical probe (OP). In group B (n = 80), the subgingival deposits were first recorded with the OP followed by root surface debridement until no subgingival deposits could be detected by the device. Teeth were then extracted and examined under a stereomicroscope by two trained dentists (DENT 1, DENT 2) and a dental student. Results were compared with the measurements of the OP and direct visual control. RESULTS: In group A, post extraction results revealed 89% and 90% agreement with the positive and negative OPs detection. In group B, 17% of the surfaces demonstrated even after subgingival debridement and control with the detecting device still calcified deposits in the microscopic evaluation. The highest inter-examiner agreements were observed between DENT 1 and DENT 2. CONCLUSION: It was shown that the optical detection system identifies subgingival calculus with a high efficacy and therefore, may be a support for the operator to determine the endpoint of root surface instrumentation. PMID- 18412729 TI - Empowering dental hygienists: tobacco cessation resources. PMID- 18412730 TI - Is your knowledge up-to-date? PMID- 18412732 TI - Psychological stress and health in undergraduate dental students: fifth year outcomes compared with first year baseline results from five European dental schools. AB - Psychological stress in undergraduate dental students: fifth year outcomes compared with first year baseline results from five European dental schools. OBJECTIVE: To compare the levels of a series of health-related indicators from a cohort of fifth year dental students from five European schools with their first year scores, and to investigate the relationship between these follow-up measures. METHODS: Burnout was measured using the Maslach Burnout Inventory (MBI), consisting of three scales: Emotional Exhaustion (EE, alpha = 0.90), Depersonalisation (alpha = 0.80) and Personal Accomplishment (alpha = 0.72). Physical health was measured by the Physical Symptoms Questionnaire (alpha = 0.82), psychological distress was measured using the General Health Questionnaire (GHQ, alpha = 0.89) and student stress was captured using seven subscales of the Dental Environment Stress questionnaire (DES, alpha = 0.92). A total of 132 fifth year students responded from five dental schools (Manchester, Belfast, Cork, Helsinki and Amsterdam), a 51% response. RESULTS: Fifth year students showed relatively high mean MBI scores when compared with first year results, especially on EE; 39% could be labelled 'high scorers'; 44% of the students met the criteria for 'cases' on the GHQ. Highest mean scores on the DES were obtained on the subscales: Study Obligations, Patient-Related Aspects and Study Pressure respectively. Between schools interesting differences were detected on all variables. As hypothesised, a clear direct effect of stress on both burnout and physical symptoms was shown. An indirect effect of stress on mental health via burnout was shown. CONCLUSIONS: Dental students showed a negative development through the years from first to fifth year with regard to EE and psychological distress. Both burnout constructs related to physical and mental health. It is recommended that dental faculty focus on the importance of prevention and intervention of stress amongst undergraduates. PMID- 18412733 TI - What becomes of dental research trainees once they leave the Dental Research Institute? An analysis over 53 years. AB - The careers of 131 dental research trainees were followed (1954-2007) to establish whether and how they utilised their research training in keeping with clinical research workforce needs. The Dental Research Institute database was used to obtain trainee demographic, teaching and research outputs which were examined according to degree types: PhD (18); MSc (55); MDent (42) and dropout (16). Current careers show that 48% are in exclusive private practice and 15% in exclusive academia with further 15% practitioners having academic links via sessional teaching or research at a dental school. Most (63%) have remained in South Africa but emigration is high amongst the PhD and MSc groups. Forty-one per cent of the cohort is of age or=14 years. MEASUREMENTS: Alcohol consumption levels associated with National Health and Medical Research Council (NHMRC) guidelines for short- and long-term harm were identified and compared with self reported measures of absenteeism due to alcohol use and due to any illness/injury. FINDINGS: More than 40% of the Australian work-force consumed alcohol at risky or high-risk levels at least occasionally. High-risk drinkers were up to 22 times more likely to be absent from work due to their alcohol use compared to low-risk drinkers. Short-term high-risk drinkers were also significantly more likely to be absent from work due to any illness or injury than employed low-risk drinkers. Young employees and males were more likely to report alcohol-related absenteeism compared to older workers and females. CONCLUSIONS: The relationship between workers' alcohol consumption patterns and absenteeism is more substantial than previously recognized or documented. Alcohol related absenteeism is not restricted to small numbers of chronic heavy drinkers, but also involves the much larger number of risky non-dependent drinkers who drink less frequently at risky levels. To improve workers' health and wellbeing and enhance productivity and economic prosperity, appropriate education, prevention and policy strategies are warranted and necessitate revision of previously narrow approaches undertaken with work-place programmes. PMID- 18412753 TI - The joint association of average volume of alcohol and binge drinking with hazardous driving behaviour and traffic crashes. AB - BACKGROUND: Previous studies on alcohol-related road safety have not assessed the joint impact of average volume of alcohol and binge drinking. AIM: To examine the joint and separate association of average volume of alcohol and binge drinking with hazardous driving behaviour and traffic crashes. METHODS: Data were drawn from telephone interviews conducted in the period 2000-2005, with 12 037 individuals representative of the population aged 18-64 years in the Madrid region, Spain. The threshold between average moderate and heavy volumes was 40 g of alcohol/day in men and 24 g/day in women. Binge drinking was defined as intake of >or= 80 g of alcohol in men and >or= 60 g in women, during any drinking occasion in the preceding 30 days. Individuals were classified into the following categories: (i) non-drinkers; (ii) moderate drinkers with no binge drinking (MDNB); (iii) moderate drinkers with binge drinking (MDB); (iv) heavy drinkers with no binge drinking (HDNB); and (v) heavy drinkers with binge drinking (HDB). Analyses were performed using logistic regression, with adjustment for sex, age and educational level. FINDINGS: Frequency of inadequate seat-belt use increased progressively across categories of alcohol consumption, with odds ratio (OR) 1 in non-drinkers, 1.19 [95% confidence interval (CI) 1.06-1.33] in MDNB, 1.69 (1.41 2.03) in MDB, 1.68 (1.24-2.29) in HDNB and 2.41 (1.83-3.18) in HDB (P for trend <0.001). Compared with MDNB, alcohol-impaired driving was also more frequent in MDB (OR 7.43; 95% CI: 5.52-10.00), HDNB (OR 7.31; 95% CI: 4.37-12.25) and in HDB (OR 15.50; 95% CI: 10.62-22.61). Lastly, compared with non-drinkers, frequency of traffic crashes increased progressively across categories of alcohol consumption (P for trend=0.028), although it only reached statistical significance in HDB (OR 2.01; 95% CI: 1.00-4.09). CONCLUSIONS: Self-reported average volume of alcohol and binge drinking are both associated with self-reported hazardous driving behaviour and traffic crashes. The strength of the association is greater when average heavy consumption and binge drinking occur jointly. PMID- 18412755 TI - Low identification of alcohol use disorders in general practice in England. AB - AIMS: The prevalence of alcohol use disorders (AUDs) in the United Kingdom is estimated at 25%, and primary care has been identified as the first line of treatment for this population. However, there is a paucity of evidence regarding the current rates of identification of AUDs in primary care. The aim of the present study was to compare the observed rates of AUDs in general practice with expected rates, which are based on general population prevalence rates of AUDs. DESIGN, PARTICIPANTS AND MEASUREMENTS: Epidemiological data on individuals aged 16-64 years with an AUD was obtained from the General Practice Research Database. General population prevalence rates of AUDs were obtained from the Psychiatric Morbidity Survey. Chi(2) tests and identification ratios were used to analyse the data. RESULTS: There was a significant relationship between type of AUD and identification (chi(2)=1466.89, P<0.001), and general practitioners were poorer at identifying harmful/hazardous drinkers when compared with dependent drinkers. No gender differences in the identification of hazardous/harmful drinking were found, but female dependent drinkers were significantly more likely to be identified than males (identification ratio 0.07; 95% confidence interval 0.06 0.07). The identification of AUDs was significantly lower for the 16-24-year age group compared with all other age groups. CONCLUSION: Despite attempts at targeting hazardous/harmful drinkers for brief interventions in primary care, the present findings suggest that this group are still under-identified. Furthermore, this under-identification is even more apparent in men and in young people who have high general population prevalence rates for AUDs. In conclusion, increasing identification rates could be incorporated into brief intervention strategies in primary care. PMID- 18412754 TI - The incidence and severity of hangover the morning after moderate alcohol intoxication. AB - AIMS: To determine the incidence and covariates of hangover following a night of moderate alcohol consumption at a targeted breath alcohol level. DESIGN: Data were combined from three randomized cross-over trials investigating the effects of heavy drinking on next-day performance. A total of 172 participants received either alcoholic beverage (mean=0.115 g% breath alcohol concentration) or placebo on one night and the other beverage a week later. The next day, participants completed a hangover scale. PARTICIPANTS: Participants were 54 professional merchant mariners attending a recertification course at Kalmar Maritime Academy (Kalmar, Sweden) and 118 university students or recent graduates recruited from greater Boston. SETTING: One trial was conducted at Kalmar Maritime Academy (Sweden); the other two were conducted at the General Clinical Research Center at Boston Medical Center. MEASUREMENTS: A nine-item scale assessed hangover. FINDINGS: Hangover was reported by 76% of participants. Neither alcoholic beverage type nor participant characteristics was associated with incidence of hangover. CONCLUSIONS: Our findings on the propensity of hangover suggest that 25 30% of drinkers may be resistant to hangover. PMID- 18412756 TI - Working memory and multi-tasking in paranoid schizophrenia with and without comorbid substance use disorder. AB - AIMS: Addiction is a frequent comorbid disorder in schizophrenia and associated with poor outcome. The present study sought to determine whether addicted and non addicted schizophrenic patients are impaired differentially on the executive abilities of working memory and multi-tasking which are relevant for maintaining abstinence. DESIGN: Comparisons of executive performance in clinical and control groups. SETTING: In-patient setting. PARTICIPANTS: The cognitive profile of schizophrenic patients with and without comorbid substance abuse disorder was compared with that of patients suffering from major depression or alcoholism and healthy participants. MEASUREMENTS: A range of cognitive tasks was used to assess: (i) the ability to update continuously context information in working memory and to use it for action selection; and (ii) the capacity to divide attention between different sensory input channels and to coordinate verbal and manual responses. FINDINGS: Single-diagnosis schizophrenic patients showed pronounced impairments on measures of online maintenance and use of context information. Their ability to coordinate different sensory input channels (divided attention) was also impaired. Addicted schizophrenics showed evidence of impaired sensory input management and of reduced context sensitivity, when age differences were controlled. CONCLUSIONS: The present study indicates severe working memory and multi-tasking deficits in schizophrenia which are, however, not exacerbated by comorbid addiction. PMID- 18412757 TI - Cannabis withdrawal predicts severity of cannabis involvement at 1-year follow-up among treated adolescents. AB - AIMS: Controversy exists regarding the inclusion of cannabis withdrawal as an indicator of dependence in the next revision of the Diagnostic and Statistical Manual of Mental Disorders (DSM) and International Classification of Diseases (ICD). This study contrasted the concurrent and predictive validity of three operational definitions of cannabis withdrawal in a sample of treated adolescents. DESIGN: Prospective study of treated adolescents with 1-year follow up. SETTING AND PARTICIPANTS: Adolescents (n=214) were recruited from intensive out-patient treatment programs for substance abuse, and followed at 1 year (92% retention). Youth who were included in the analyses reported regular cannabis use. MEASUREMENTS: The number of DSM-IV cannabis abuse and dependence symptoms at baseline and 1-year follow-up, past year frequency of cannabis use at baseline and follow-up, and periods of abstinence at 1-year follow-up. Cannabis withdrawal was defined based on (i) the presence of two or more cannabis withdrawal symptoms; (ii) a definition proposed by Budney and colleagues (2006) that requires four or more withdrawal symptoms (four-symptom definition); and (iii) the use of latent class analysis to identify subgroups with similar cannabis withdrawal symptom profiles. FINDINGS AND CONCLUSIONS: All three definitions of cannabis withdrawal demonstrated some concurrent validity. Only the four-symptom and latent class-derived definitions of withdrawal predicted severity of cannabis related problems at 1-year follow-up. No cannabis withdrawal definition predicted frequency of use at follow-up. Further research is needed to determine the clinical utility and validity of the four-symptom definition, as well as alternative definitions of cannabis withdrawal, to inform revisions leading to DSM-V and ICD-11. PMID- 18412758 TI - Longitudinal changes in methamphetamine and cocaine use in untreated rural stimulant users: racial differences and the impact of methamphetamine legislation. AB - AIMS: To examine how race and methamphetamine legislation are associated with changes in cocaine and methamphetamine use among untreated rural stimulant users. DESIGN: A longitudinal study of stimulant users identified through respondent driven sampling. SETTING: Rural areas of three US states. PARTICIPANTS: Participants at baseline were current users of methamphetamine and/or cocaine. MEASURES: Self-reports of methamphetamine, crack cocaine and powder cocaine use were assessed at 6-month intervals over a 2-year period. Generalized estimating equations were performed to account for correlations between the repeated measurements. FINDINGS: Compared to whites, African Americans were much more likely to use crack cocaine, equally likely to use powder cocaine and much less likely to use methamphetamine. Both whites and African Americans reduced their consumption of methamphetamine and both forms of cocaine over 2 years. Exposure to laws restricting the purchase of over-the-counter cold medications containing methamphetamine precursors was not associated with methamphetamine use, but associated with a slight rise in powder and crack cocaine use. CONCLUSIONS: The study yielded disconcerting as well as promising findings regarding the natural history of stimulant use in rural areas. Of some concern is that methamphetamine precursor laws were correlated with increased cocaine consumption, diminishing their net public health benefits. However, despite its insurgence in rural areas of the United States, very few African Americans have initiated methamphetamine use. Regardless of race, many stimulant users stopped using cocaine and methamphetamine without formal substance abuse treatment over 24 months. PMID- 18412760 TI - How when matters for quitting and relapse. PMID- 18412759 TI - Menstrual phase effects on smoking relapse. AB - AIMS: To examine if menstrual phase affects relapse in women attempting to quit smoking. DESIGN: An intent-to-treat randomized smoking cessation trial where women were assigned to quit smoking in either the follicular (F) or luteal (L) menstrual phase and were followed for up to 26 weeks. They were assessed for relapse by days to relapse and relapse phase to determine if those who begin a quit attempt during the F phase were more successful than those who begin during the L phase. SETTING: Tobacco Use Research Center, University of Minnesota, Minneapolis, Minnesota. PARTICIPANTS: A total of 202 women. MEASUREMENTS: Latency to relapse from continuous and prolonged abstinence, point prevalence, phase of relapse, first slip within the first 3 and 5 days post-quit date, subject completion rates and symptomatology (i.e. withdrawal and craving). FINDINGS: The mean days to relapse from continuous abstinence and relapse from prolonged abstinence for the F group were 13.9 and 20.6 days, respectively, and 21.5 and 39.2 days, respectively, for the L group. Using point prevalence analysis at 14 days, 84% of the F group had relapsed compared with 65% of the L group [chi(2)=10.024, P=0.002; odds ratio (OR) = 2.871, 95% confidence interval (CI), 1.474-5.590]. At 30 days, 86% of the F group relapsed, compared with 66% of the L group (chi(2)=11.076, P=0.001; OR=3.178, 95% CI, 1.594-6.334). CONCLUSION: Women attempting to quit smoking in the F phase had less favorable outcomes than those attempting to quit in the L phase. This could relate to ovarian hormones, which may play a role in smoking cessation for women. PMID- 18412761 TI - The role of national policies intended to regulate adolescent smoking in explaining the prevalence of daily smoking: a study of adolescents from 27 European countries. AB - AIMS: This study seeks to examine whether contextual factors influence adolescents' daily smoking. A focus was placed on three modifiable policies operating at a national level, non-smoking policy at educational facilities, price and minimum age for buying tobacco. DESIGN: This study is based on a merged data set consisting of the 2001/02 Health Behavior in School-aged Children (HBSC) study and national-level data collected from the 2003 WHO European Tobacco Control Database and the World Development Indicators Database. HBSC is an international study including adolescents from 32 countries in Europe, Israel and North America. Data were analysed with multi-level hierarchical regression models. FINDINGS: The study found large differences in the prevalence of daily smoking among adolescents, and also large differences between boys and girls within some countries. The study found that smoking bans in schools were associated with lower odds ratios of daily smoking, which was the one positive association in the study. The study found no association between cigarette prices and adolescent daily smoking prevalence, and also the somewhat unexpected finding that having an age limit for allowing adolescents to purchase tobacco was associated with an increased risk of daily smoking. CONCLUSIONS: There was an association between mandatory national bans on smoking and lower smoking prevalence. This should be confirmed by studies that examine whether mandatory bans are more rigorously implemented than voluntary bans. If this association is causal, introducing mandatory bans may reduce adolescent smoking prevalence. The findings that price was unrelated to smoking prevalence undermine findings elsewhere that adolescent smokers are more price-sensitive than adult smokers, but longitudinal studies are needed. PMID- 18412762 TI - The problems of making policy. PMID- 18412763 TI - The cost-effectiveness of a smoking cessation program for out-patients in treatment for depression. AB - AIMS: Smoking cessation programs are highly cost-effective. The cost effectiveness of programs for psychiatric patients may be affected by differences in cost, efficacy, survival and quality of life. We evaluated cost-effectiveness of a program for smokers being treated for depression. DESIGN: A randomized trial compared brief contact to a stepped smoking cessation program in 322 cigarette smoking mental health out-patients. We determined the intervention's direct cost. Because smoking cessation may affect short-term use of mental health care, we used administrative databases and self-report to find mental health-care cost. FINDINGS: The 163 individuals randomized to stepped care received an average of $346 of smoking cessation services, including $221 for computer-mediated assessments of readiness to quit, and $124 for counseling, nicotine replacement therapy and bupropion. The cessation program was used by 53 participants. Total cost of smoking cessation and mental health services was $4805 in the stepped care group and $4173 in the brief-contact care group (not significantly different). After 18 months of follow-up, the stepped care group had 5.5% greater abstinence from smoking. Smoking cessation services cost $6204 per successful quit. Cessation services and mental health care cost was $11 496 per successful quit. CONCLUSION: If smoking cessation yields 1.2 years additional life, the cessation services cost $5170 per life-year, and cessation services and mental health care cost $9580 per life-year. Even if quitting does not increase survival of depressed individuals as well as in other smokers, the stepped care intervention is likely to be regarded as cost-effective. PMID- 18412764 TI - A comparison of the Fagerstrom Test for Nicotine Dependence and smoking prevalence across countries. AB - AIMS: To examine the correlation between the Fagerstrom Test for Nicotine Dependence (FTND) score and smoking prevalence across countries. DESIGN: Cross sectional study. SETTING: Fifteen studies from 13 countries with FTND score data. PARTICIPANTS: Samples of smokers were identified through systematic literature searches, web queries and colleagues. Smokers were considered representative of their country's smoking population if they were drawn from population-based sources, were not seeking smoking cessation treatment and did not have significant comorbidities. Smoking prevalence data were derived from the study itself or the country's population rate of daily smoking for the study year. MEASUREMENTS: A Pearson correlation coefficient was used to examine the direction and magnitude of the correlation between FTND score and smoking prevalence across countries. FINDINGS: FTND scores ranged from 2.8 to 4.6. Smokers in Germany and Norway had the lowest FTND scores, while smokers in Sweden and the United States had the highest FTND scores. The prevalence of daily smoking in these countries was very different: 37% and 30% in Germany and Norway, 19% and 16% in the United States and Sweden, respectively. An inverse correlation towards higher FTND scores in countries with lower smoking prevalence was found (r=-0.73, P=0.001). Current smokers had higher FTND scores than former smokers. CONCLUSIONS: The significant inverse correlation between FTND score and smoking prevalence across countries and higher FTND score among current smokers supports the idea that remaining smokers may be hardening. Less dependent smokers may quit more easily and remaining dependent smokers may need more intensive treatment. PMID- 18412765 TI - Parental smoking and smoking experimentation in childhood increase the risk of being a smoker 20 years later: the Childhood Determinants of Adult Health Study. AB - AIMS: To examine the long-term effects of childhood smoking experimentation and exposure to parental smoking on adult smoking risk. METHODS: Data were from a 20 year follow-up of 9-15-year-olds who completed questionnaires in the 1985 Australian Schools Health and Fitness Survey (n=6559). The relative risks (RR) of adult current smoking in 2004-05 for childhood exposure to smoking experimentation (never, a few puffs, <10 cigarettes, >10 cigarettes) and parental smoking (none, father, mother, both parents) in 1985, with adjustment for confounders, were estimated by log binomial modelling. Analyses were stratified by age (9-13 and 14-15 years) and sex. FINDINGS: Participation at follow-up was 54% (n=3559). Childhood smoking experimentation increased the risk of being a current smoker particularly for 14-15-year-old experimenters of more than 10 cigarettes [males, RR 2.72, 95% confidence interval (CI) 1.74-4.25; females, RR 6.39, 95% CI 2.85-14.33]. Parental smoking was associated with adult current smoking risk, particularly for 9-13-year-olds with two smoking parents (males, RR 1.53, 95% CI 1.19-1.96; females, RR 1.99, 95% CI 1.52-2.61) and older males with smoking mothers (RR 1.82, 95% CI 1.22-2.73). Parental smoking was not associated with childhood smoking experimentation. CONCLUSIONS: These findings suggest that any childhood smoking experimentation increases the risk of being a smoker 20 years later. As exposure to parental smoking predicted current smoking, parents should be aware of the association between their own smoking behaviour and that of their children. PMID- 18412766 TI - Ethics of alcohol policy in Brazil: why it is possible to be independent when we sit at the same table with the alcohol industry. PMID- 18412767 TI - No one can serve two masters. PMID- 18412768 TI - Response to Dr Kari Poikolainen: The persistent, alternative argument to apparent cardioprotective effects of alcohol. PMID- 18412773 TI - Pregnancy in uremic patients: an eventful journey. AB - Renal damage, which can be caused by gestational anomalies such as pre-eclampsia, carries a risk of gestational complications; the greatest risk being in women who become pregnant while on hemodialysis or peritoneal dialysis. If this rare event occurs, there is a marked increase in the risk of pre-eclampsia, early uterine contractions and hydramnios, hypertensive crisis, preterm delivery and intrauterine growth retard. Furthermore, newborns are almost always of low birthweight. Patients who undergo renal transplantation are another high-risk category. In such cases, the pregnancy itself can compromise the success of the transplant and the immunosuppressive therapy correlated to it can become a threat to the course of the pregnancy and normal fetal growth. Therefore, in view of the lack of appropriate guidelines for the best possible approach to the treatment of women on dialysis or of those with a renal transplantation, it is best to advise these patients against becoming pregnant and/or to provide a valid counselling service illustrating the extreme difficulties and dangers involved in becoming pregnant. PMID- 18412772 TI - An integrated view of L-selectin and trophinin function in human embryo implantation. AB - Determining molecular mechanisms of human embryo implantation is an extremely challenging task due to the limitation of materials and significant differences underlying this process among mammalian species. Recently, L-selectin and its ligand carbohydrate have been proposed as a system that mediates initial adhesion of human blastocysts to the uterine epithelia. We have also identified trophinin as a unique apical cell adhesion molecule potentially involved in the initial adhesion of trophectoderm of the human blastocyst to endometrial surface epithelia. In the mouse, the binding between ErbB4 on the blastocyst and heparin binding epidermal growth factor-like growth factor on the endometrial surface enables the initial step of the blastocyst implantation. The evidence suggests that L-selectin and trophinin are included in human embryo implantation. This review summarizes findings relevant to the functions of L-selectin and trophinin in human embryo implantation, and proposes a model that reconciles these cell adhesion mechanisms. PMID- 18412774 TI - Stimuli through Toll-like receptor (TLR) 3 and 9 affect human chorionic gonadotropin (hCG) production in a choriocarcinoma cell line. AB - AIM: Toll-like receptors (TLR) are expressed mostly on mucosal innate immune cells which recognize microorganism-associated structures, the so called pathogen associated molecular patterns (PAMP). Recently, possible roles of TLR in placental tissue, especially their role in protecting the conceptus from maternal infection, have been discussed, but the physiopathological roles of TLR in placental functions have yet to be examined. METHODS: The choriocarcinoma cell line BeWo was used as a model of trophoblast stem cells. First, the expression of TLR 1-9 on BeWo cells was investigated using reverse transcriptase-polymerase chain reaction. Then, the cells were treated with TLR 1-9 agonists, using the human TLR 1-9 agonist kit (Pam3CKS4, HKLM, Poly (I:C), Escherichia coli K12 lipopolysaccharide (LPS), Salmonella typhimurium flagellin, FSL1, imiquimod, ssRNA 40, and ODN 2006). The concentration of human chorionic gonadotropin (hCG) in the cell culture supernatants was examined on ELISA. RESULTS: BeWo cells constitutively expressed TLR 1-9 mRNA in the presence or absence of forskolin. The TLR 3 agonist, Poly(I:C), and the TLR 9 agonist, CpG-DNA (ODN2006), upregulated hCG production but had minimal effects in the absence of forskolin. Stimulation with the other TLR agonists produced no remarkable increase, whether in the presence or absence of forskolin. CONCLUSIONS: TLR 3 and TLR 9 agonists upregulated hCG production in BeWo cells. The present observations suggest alternative roles of TLR signaling in the control of placental functions and blastocyst-endometrial dialogue, as well as in pathophysiology, including for infertility and ectopic pregnancy. PMID- 18412775 TI - Comparison of cell cycle regulatory gene mRNA in three different types of human trophoblasts and effect of transforming growth factor. AB - AIM: Identifying the factors responsible for reducing the proliferation, syncytialization, and invasiveness of trophoblast tissues, as seen with preeclampsia, intrauterine growth restriction, and spontaneous miscarriage, is a current challenge in reproductive biology. These factors, transforming growth factor (TGF)-beta as an example, can work by altering trophoblast differentiation or proliferation. We therefore investigated and compared specific markers of trophoblast proliferation and differentiation in three commonly used trophoblast tissue cell models, and also investigated the influence of TGF-beta on these markers. METHODS: In this study, we isolated human trophoblasts from first trimester and term placentas, and additionally used human choriocarcinoma cells (JEG-3). Baseline values of human chorionic gonadotropin (hCG) secretion and relative mRNA levels of cell cycle regulators (cyclin E, p21, p27, and p57) were investigated for each cell type. We also investigated the influence of TGF-beta on these parameters. RESULTS: Quantitative and longitudinal production of hCG differed between the three cell types. Significantly different amounts of cyclin E, p21, p27, and p57 mRNA were demonstrated within each cell type, as well as between all the cell types, throughout the culture time period. Each trophoblast type demonstrated a reduction of hCG secretion in response to TGF-beta. TGF-beta did not show a consistent effect on the cell cycle mRNA of any of the cell types. CONCLUSION: We were able to characterize and compare the differential production of hCG, as well as the differential expression of cell cycle-associated mRNA of early trophoblasts, term trophoblasts, and choriocarcinoma cells. The production of hCG was altered by TGF-beta, although mRNA levels were not markedly altered by TGF-beta. PMID- 18412776 TI - Perinatal magnetic resonance fetal lung volumetry and fetal lung-to-liver signal intensity ratio for predicting short outcome in isolated congenital diaphragmatic hernia and cystic adenomatoid malformation of the lung. AB - AIM: To evaluate the usefulness of magnetic resonance imaging (MRI) in predicting the risk of lethal pulmonary hypoplasia in fetuses. METHODS: The subjects consisted of 15 fetuses (29-40 weeks' gestation), including fetuses with major malformation diagnosed on prenatal ultrasonography. MRI using a 1.5 T magnet and half-Fourier acquisition single-shot fast spin-echo sequences were applied to all fetuses at 29-36 weeks. Fetal lung-to-liver signal intensity ratio (LLSIR) was calculated by medians of region-of-interest analysis; estimated fetal bodyweight (FBW), by ultrasonography; and estimated fetal lung volume (FLV), by planimetric measurement of total lung volume. FLV/FBW was also calculated. The presence of the pulmonary hypoplasia in neonates was identified based on clinical and anatomico-pathological findings. Differences in LLSIR and FLV/FBW were analyzed for surviving and non-surviving neonates. RESULTS: Ten surviving neonates had a median LLSIR of 3.00, range: 1.60-4.40, while that in seven non-surviving neonates was 2.21, range: 0.70-3.72; no significant difference was found between the groups. Surviving neonates had a median FLV/FBW of 11.4, range: 7.1-15.7, while that in non-surviving neonates was 4.4, range: 3.1-5.7. FLV/FBW in non surviving neonates was significantly lower than that of the FLV/FBW for surviving fetuses (P<0.05). CONCLUSIONS: Low FLV/FBW may be useful in prenatally predicting mortality in fetuses with pulmonary hypoplasia. PMID- 18412777 TI - Positive rates for rubella antibody in pregnant women and benefit of post-partum vaccination in a Japanese perinatal center. AB - AIM: This study of pregnant women was undertaken to clarify their immune status for rubella and to evaluate the efficacy of, and adverse reaction to, post-partum rubella vaccination. METHODS: We determined the levels of both rubella antibody by hemagglutination inhibition test and IgM antibody by enzyme-linked immunosorbent assay in the first gestational trimester and surveyed the positive rates for rubella antibody in pregnant women at our perinatal center; we recommended post-partum vaccination for women with poor immune status. We also evaluated the effectiveness of the vaccine and any adverse reaction. RESULTS: Among 2741 pregnant women, 185 cases (6.7%) were seronegative for rubella virus. This rate was especially high (12.4%) in the population who had opted out of the vaccination because of a change in Japanese government policy. A total of 30 cases (1.1%) were positive or false-positive for IgM antibody, but no congenital rubella syndrome (CRS) developed. The seropositive rate was 93.3% in total, but this figure includes those who required post-partum vaccination (558 cases or 20.3%) because their titers were 16x or lower. Among 145 cases who received post partum rubella vaccine, no severe adverse reaction was detected, and in all 37 cases with hemagglutination inhibition antibody titers of 8x or lower, the efficacy rate was 100%. CONCLUSIONS: It is necessary to appropriately evaluate the CRS risk. Also, we need to emphasize the importance and safety of post-partum rubella vaccination in Japan. PMID- 18412778 TI - Thrombophilic mutations in pre-eclampsia and pregnancy-induced hypertension. AB - AIM: The aim of the present study was to determine the existence or prevalence of thrombophilic markers such as Factor V Leiden, prothrombin G20210A, protein S, protein C, activated protein C and anti-thrombin in pre-eclampsia and pregnancy induced hypertensive patients. METHODS: Blood samples were collected from a total number of 124 women at the maternity unit, University of Malaya Medical Center. These included 49 patients with pre-eclampsia, 63 patients with pregnancy-induced hypertension and 12 normal pregnant women. DNA was extracted from the blood samples. Factor V Leiden (Taq I) and prothrombin G20210A (Hind III) genotyping was done on polymerase chain reaction-restriction fragment length polymorphism. Anti-thrombin activity and the concentrations of protein C, protein S and activated protein C were measured using the IL Coagulation System (Hemosil). RESULTS: Of the 124 subjects, one pre-eclampsia patient was homozygous for Factor V Leiden mutation but prothrombin G20210A mutation was not present in any of the subjects. The subject with Factor V Leiden mutation also had a low activated protein C resistance and a low protein S concentration. CONCLUSIONS: Factor V Leiden mutation is present in the Asian population and may very well serve as one of the genetic factors responsible for pre-eclampsia and other adverse pregnancy outcomes. PMID- 18412779 TI - Pre-eclampsia outcomes in different hemodynamic models. AB - AIM: To evaluate whether there is a significant relationship between hemodynamic models and pre-eclampsia outcomes. METHOD: A controlled experimental study was performed. We analyzed 2910 hemodynamic series systematically sampled from 970 pregnant women three times every 2 weeks from the definite diagnosis of pre eclampsia until delivery. Women were divided into three groups based on total peripheral resistance (TPR): a low-TPR group, a normal-TPR group and a high-TPR group. Every group was divided into three subgroups based on cardiac index (CI): a low-CI subgroup, a normal-CI group and high-CI group. Common lab tests, electrocardiographic examination, fundus examination, cardiac function, liver function and kidney function were measured after every hemodynamic monitoring. Primary outcomes included various maternal and neonatal morbidity and neonatal and infant mortality. RESULTS: In our study we found seven hemodynamic models in pre-eclampsia during the third trimester of pregnancy. No significant differences in maternal age, weight, and height were observed between the three groups. The low-TPR and normal-TPR groups showed better disease results than the high-TPR group in respect of HELLP (P<0.01), lung edema (P<0.01), acute renal failure (P<0.01), heart failure (P<0.01), neonatal intensive care unit admission (P<0.01), infant weight (P<0.01), neonatal mortality (P<0.01) and infant mortality (P<0.01). The low-TPR group had better results than the normal-TPR group in respect of eclampsia (P<0.01), liver hemorrhage (P<0.01), birthweight (P<0.01) and gestational week at birth (P<0.05). CONCLUSION: Among all the subgroups, the highest maternal and neonatal morbidity was in the high-TPR-high CI subgroup. There is clear relationship between hemodynamic and disease outcomes during the third trimester. PMID- 18412780 TI - Safety and usefulness of emergency maternal transport using helicopter. AB - AIM: Japan has a shortage of tertiary medical care facilities for maternal and fetal medicine. Establishment of efficient medical transport systems is needed for pregnant women and fetuses with severe complications. Maternal transport by helicopters is expected to shorten transportation time to advanced facilities, although its feasibility has not yet been evaluated. The aim of the present study was to investigate the status of maternal helicopter transport, and conditions of the pregnant patients and children transferred by helicopter to Kameda Medical Center (KMC). METHODS: Between August 2005 and July 2006, 26 pregnant women were transported by helicopters to KMC. RESULTS: The median net flight time was 24 min (range 15-29 min), and the median of estimation of ground transportation time was 125 min (range 90-180 min). The causes for transfers were preterm labor in eight, preterm premature rupture of the membrane in five, cervical incompetence in five, pre-eclampsia in three and other medical reasons in five. Five of the 26 patients were discharged with restored stability of pregnancy. The remaining 21 patients underwent delivery at KMC. The median gestational age was 26 weeks (range 22-33 weeks) at the time of transfer and 31 weeks (range 22-37 weeks) at delivery. Four of 26 neonates who were born at KMC died after birth due to severe premature or congenital anomaly. Seventeen of the remaining 22 neonates, including 10 twins, received treatment in the neonatal intensive care unit. All of the 22 neonates and all the mothers were discharged in good condition. No patients developed any complications requiring treatment during flights. CONCLUSION: Helicopter transfer is feasible for pregnant patients with severe complications. PMID- 18412781 TI - Adolescent primiparas: changes in obstetrical risk between 1983-1987 and 1999 2005. AB - AIMS: Teenage pregnancies have always been considered at increased risk for obstetric complications. Deliveries in adolescent primiparas in the 5-year time periods 1983-1987 and 1999-2005 were compared against each other, the general population and against primiparas aged 20-29 years in order to reveal trends and differences in obstetric outcome. METHODS: A total of 186 primiparas delivering at an age of 17 or less between October 1999 and October 2005 were compared with 353 adolescent primiparas delivered between 1983 and 1987. Type of delivery and complications such as low birthweight, pre-eclampsia, breech presentation and third stage complications were studied. RESULTS: The percentage of adolescents in the overall obstetric population decreased. The cesarean section rate remained the same in the adolescents while increasing in the general population. Rates of low birthweight and operative vaginal delivery increased in the adolescent group and overall. Third stage complications (abnormally adherent or incomplete placentas) decreased in both groups. There were no intrauterine fetal deaths in adolescent pregnancies in either time period. Other obstetric variables were unchanged in the adolescent as well as in the general population between 1999 and 2005. When comparing the adolescents' outcome with the outcome of the 20-29-year old primiparas between 1999 and 2005, it was noted that the rates of abstracted obstetric variables were higher in the population of the 20-29-year-olds. CONCLUSIONS: The obstetric outcome of adolescent pregnancies has remained favorable over the last 18 years. We do not consider adolescence as an obstetrical risk. We suggest that adolescent pregnancy is more a public health issue than a clinical problem. PMID- 18412782 TI - Randomized comparison of dry tablet insertion versus gel form of vaginal misoprostol for second trimester pregnancy termination. AB - AIM: To compare the effectiveness of vaginal misoprostol between dry tablet insertion and gel form for second trimester pregnancy termination. METHODS: A non blinded block randomized controlled trial was conducted on 148 pregnant women with live fetuses in the second trimester undergoing pregnancy termination. They were randomly allocated to receive vaginal misoprostol (400 microg) either dry tablet insertion (n=72) or gel form (n=76). The same dose was then repeated every 3 h if adequate uterine contraction was not achieved until 48 h after the initiation of misoprostol. If abortion did not occur within this period, the treatment was considered a failure and other technique of termination was then given based on the decision of the attending physicians and the cervical status. RESULTS: The mean induction-abortion interval in group 1 (20.9+/-12.3 h) was not significantly different from that in group 2 (17.7+/-10.2 h). The mean total dose of misoprostol was also not significantly different between the two groups (group 1, 1556.9 microg; group 2, 1350.9 microg), but the adverse effects of misoprostol (chill and diarrhoea) were more common in the gel group. CONCLUSION: Tablet insertion or gel form of vaginal misoprostol have similar effectiveness but the gel form was associated with more common adverse effects. PMID- 18412783 TI - Underlying pathology of women with "atypical squamous cells, cannot exclude high grade squamous intraepithelial lesion" smears, in a region with a high incidence of cervical cancer. AB - AIM: To evaluate the histopathology of women who had "atypical squamous cells, cannot exclude high-grade squamous intraepithelial lesions" (ASC-H) on cervical cytology in a region with high incidence of cervical cancer. METHODS: This study was conducted at Chiang Mai University Hospital, Chiang Mai, Thailand. All women with ASC-H, who had undergone colposcopic and histolopathologic evaluation between October 2004 and January 2007, were recruited. Similar cohorts with other squamous cell abnormalities on a Pap-smear, who had undergone colposcopy during the same period, were included as comparative groups. RESULTS: During the study period, 85 women who had ASC-H smears underwent colposcopic and histopathologic evaluation. The mean age was 45.3 years (range, 20-64 years). The histopathologic results of these 85 women were as follows: cervical intraepithelial neoplasia (CIN) II-III, 52 (61.2%); invasive cancer, 7 (8.2%); CIN I, 6 (7.1%); and no lesions, 20 (23.5%). The incidence of underlying CIN II or higher in an ASC-H smear (69.4%) was intermediate between atypical squamous cell of undetermined significance (22.7%), low-grade squamous intraepithelial lesion (44.7%) and high grade squamous intraepithelial lesion (90.5%) smears. There was no statistically significant difference in the incidence of CIN II or higher between women who were 40 years old or more and those who were younger (68.7% and 71.4%, respectively, P=0.81), or between pre-menopausal and post-menopausal women (71.4% and 63.6%, respectively, P=0.49). CONCLUSION: Reporting ASC-H cytology in our population is strongly associated with significant cervical pathology, particularly invasive cancer that is possibly at a rate higher than previously reported. Women who have ASC-H smears should therefore be referred for immediate colposcopy regardless of age and menopausal status. PMID- 18412784 TI - Role of human papillomavirus in the development of epithelial ovarian neoplasms in Italian women. AB - AIM: In this article, for the first time, we investigate the presence of human papillomavirus (HPV) DNA in ovarian epithelial neoplasms from a group of 71 Italian women. The follow ups of the patients with or without HPV DNA were also considered to evaluate whether HPV DNA in these tumors could be an indicator of prognosis. METHODS: HPV DNA was evaluated by polymerase chain reaction (PCR). Additionally P16 immunoreactivity was evaluated in all positive cases to demonstrate the presence of HPV. RESULTS: Only three cases out of 71 epithelial ovarian neoplasms (4.22%) in this series were weakly HPV positive. The presence of HPV DNA in these cases did not seem to be related with the type of neoplasm, their grade of differentiation, staging of development, patient's age or survival. CONCLUSION: The presence of HPV could be an expression of a latent infection. In both positive and negative HPV ovarian neoplasms, cervix showed only chronic cervicitis and no signal for HPV DNA on PCR analysis. PMID- 18412785 TI - Surgical adhesives and tubal sterilization: an experimental study. AB - AIM: Our aim was to assess the application of three currently used surgical adhesives in the tubal lumen of rabbits, to promote sterilization, using a transvaginal approach. METHODS: Fifty-seven female albino New Zealand rabbits (114 uterine tubes), which became pregnant and delivered before the experiment, were divided into four groups: GS (sham-24 tubes), GEFIBRI (0.25 mL of fibrin adhesive in 30 tubes), GE-GRF (0.25 mL of resorcin adhesive in 30 tubes) and GEBUTYL (0.25 mL of n-butyl-2-cyanoacrylate adhesive in 30 tubes). The animals were mated with proven fertile males after the experiment and observed over 30, 90 and 180 days. Pregnancy and patency were macroscopically evaluated. The tubal diameter, tubal mucosa, myosalpinx, total optical density and inflammatory process were microscopically evaluated. The statistical analysis was performed by McNemar and Wilcoxon tests for the subgroups, and Fisher's exact test and Kruskal Wallis test for the groups, the differences identified by Dunn's multiple comparisons test (P=5%). RESULTS: GS showed patency and pregnancies in all subgroups. GEFIBRI showed patency and pregnancies in all subgroups. GE-GRF did not show patency or pregnancies, but was associated with severe inflammatory process and tubal morphology alterations. GEBUTYL did not show patency, pregnancies or morphological tubal mucosa alterations. CONCLUSIONS: The n-butyl-2 cyanoacrylate adhesive effectively promoted tubal obstruction, did not cause tubal morphological alterations, nor did it impair the rabbit pregnancy. The fibrin adhesive failed to cause the occlusion. The GRF adhesive, in spite of producing tubal occlusion, caused severe uterine tubes damage. PMID- 18412786 TI - Abnormal glucose tolerance in polycystic ovary syndrome. AB - AIM: To investigate the incidence of abnormal glucose tolerance (AGT) in women with polycystic ovary syndrome (PCOS), and the reliability of fasting plasma glucose level (FPG) as a screening test for this metabolic abnormality. METHODS: Two hundred and three women (62 adolescents and 141 adults) diagnosed with PCOS underwent estimation of FPG and 2-h plasma glucose after ingestion of 75 g of glucose on an empty stomach. In each case, body mass index (BMI) was calculated from height and weight measurements. RESULTS: AGT was found in 16.3% of cases (33/203). AGT was found in 9.7% of adolescent cases (6/62) and 19.1% of adults (27/141) (P=0.03). Women with abnormal glucose tolerance had higher BMI than those with normal glucose tolerance in the two subgroups (not statistically significant). Difference in BMI between adults and adolescents with normal glucose tolerance was statistically significant, but not in cases with AGT. CONCLUSION: AGT was found in 16.3% of women with PCOS (19.1% of adults and 9.7% of adolescents). BMI alone is not a significant contributory factor causing deterioration of glucose tolerance as a woman with PCOS matures. Other clinical parameters of obesity such as upper body obesity need to be studied. All women with PCOS should undergo periodic screening for abnormal glucose tolerance. Fasting plasma glucose is a poor screening test to detect abnormal glucose tolerance. PMID- 18412788 TI - Sexual violence among pregnant women in India. AB - AIM: To discover the extent of sexual violence during pregnancy, its correlation to various variables and effects, and to consider future possibilities for creating awareness in health providers, health seekers and communities for prevention. METHODS: Two thousand pregnant women were interviewed using a semi structured, pre-designed, pre-tested questionnaire with some open-ended questions in the local language. RESULTS: Of 2000 women, 1959 (97.95%) reported that they had had sex with their partners during the current pregnancy. Of the 2000, 615 (30.7%) women had not wished to have sex during the pregnancy, but had been forced to do so. CONCLUSION: Women suffer with recurrent marital rape during pregnancy with considerable after effects. This sexual violence takes place irrespective of education or socioeconomic status. PMID- 18412787 TI - Trichomonas vaginalis infection: risk indicators among women attending for routine gynecologic examination. AB - OBJECTIVES: Trichomonas vaginalis is one of the most common agents of sexually transmitted disease, but trichomoniasis is still considered as an infection of minor importance. The aim of this study was to investigate the risk indicators for trichomoniasis in women attending for routine gynecologic examination. METHODS: The study was conducted comparing demographic and behavioral characteristics in two groups of 122 women with T. vaginalis infection and 853 uninfected women. RESULTS: Univariate analysis showed a significant increase of T. vaginalis infection among women with older age and multiple sexual partners. In multiple logistic regression analysis, the higher number of lifetime partners and older age remained significantly associated with T. vaginalis infection. CONCLUSIONS: This study suggests that T. vaginalis infection might be mainly related to lifestyle risk factors. Therefore, in our opinion, the diagnosis and treatment of this disease should be encouraged on routine gynecologic examination. PMID- 18412789 TI - Oligohydramnios and pulmonary hypoplasia: a case in which involvement of an angiotensin II receptor antagonist was suspected. AB - Administration of an angiotensin II receptor antagonist (ARB) during the second trimester of pregnancy is known to cause irreversible renal damage in the fetus. We report a case in which ARB was given to the mother from the first trimester until 26 weeks' gestation. The patient had diabetic nephropathy with accompanying nephrotic syndrome. At 8 weeks' gestation, she was started on candesartan cilexetil (an ARB). At 26 weeks' gestation, she was transferred to our center. Severe oligohydramnios was noted. The pregnancy was terminated, and she delivered at 27 weeks' gestation. The neonate weighed 884 g and died 1 h after birth. Autopsy revealed that the lung/bodyweight ratio was 0.0096 (>0.015) and pulmonary hypoplasia was noted. Histological examination of the kidneys showed tubular dysgenesis with poor differentiation of the proximal tubules. PMID- 18412790 TI - Surviving 27 weeks fetus expelled out of the ruptured rudimentary horn and detected a month later as a secondary abdominal pregnancy. AB - A pregnant woman, gravida 3 with two living children, who frequently experienced syncope from 23(+5) weeks of pregnancy onwards and recurring every week for a period of 3 weeks, was repeatedly treated in line for a case of acid peptic disease/appendicitis in various peripheral hospitals of Nepal, until ultrasonogram/magnetic resonance imaging diagnosis of an (undisturbed) live 27(+5) weeks abdominal pregnancy was made at our hospital. On laparotomy, this materialized to be secondary to the rupture of a left rudimentary horn pregnancy (evidenced from its sealed margin) which still retained a complete placenta, from where an umbilical cord was seen, traversing across towards the right side of the abdominal cavity just below the liver, securing its attachment to the surviving fetus and enclosed in an intact amniotic sac. Excision of the rudimentary horn containing the placenta was accomplished, after the delivery of a live baby weighing 650 g who unfortunately died on the third day of life. PMID- 18412791 TI - Antibiotic prophylaxis in cesarean section causing anaphylaxis and intrauterine fetal death. AB - Intrauterine fetal death and maternal shock occurred as a result of a type-1 hypersensitivity reaction following antibiotic prophylaxis in a cesarean section. Amniotic fluid embolism may mimic the condition. The ability to diagnose and treat such an event as early as possible is necessary in all maternity centers. The selection of antibiotic regimen and the type of anesthesia should be individualized depending upon the existing facilities and the patient's profile, especially in a resource-scarce developing country. PMID- 18412792 TI - Umbilical cord strangulation by an amniotic band resulting in a stillbirth. AB - BACKGROUND: Amniotic band syndrome with umbilical cord strangulation is extremely rare and is usually described during second trimester. We present a case of umbilical cord strangulation causing fetal demise in a full-term otherwise healthy fetus. CASE: A 39-year-old gravida 15 para 12 after one previous cesarean section presented with reduced fetal movements at the 41st gestational week. On admission, fetal heart tracing, biophysical profile and oxytocin challenge test were normal. Later, an intrauterine fetal demise was diagnosed. After delivery, an amniotic band causing strangulation of the umbilical cord was observed. The fetus was without apparent anomalies. CONCLUSION: Although extremely rare, constriction of the umbilical cord by an amniotic band can cause its strangulation followed by a stillbirth even in full-term otherwise healthy fetus. PMID- 18412793 TI - Abruptio placentae and fetal death following a Malayan pit viper bite. AB - Reports of venomous snakebites during pregnancy are uncommon. Little is known about maternal and fetal outcome following the venomous snakebite of a pregnant woman, and there is no consensus for proper management. In southern Thailand, Malayan pit viper (Calloselasma rhodostoma) bites are quite common, and we have some experience with bites of pregnant women. With these victims, the toxicity is most severely expressed in a bleeding disorder that is a significant cause of both morbidity and mortality in both the gravid woman and the fetus. Herein, because there are few such published reports, we report the case of a 43-year-old woman, gravida 5 para 4, 32 weeks pregnant who was bitten by a Malayan pit viper and, as a result, developed abruptio placentae, coagulopathy, and death of fetus in utero. She otherwise responded well to antivenom and blood components. A hysterotomy was performed and the postoperative course was unremarkable. PMID- 18412794 TI - Prenatal diagnosis and management of a ranula. AB - A ranula is a mucous extravasation cyst which appears as a swelling in the submental and submandibular regions. We describe a case of congenital ranula or retention salivary cyst diagnosed at 22 weeks' gestation. The fetal growth was normal, as was the amniotic fluid volume. Follow-up ultrasound scans revealed no changes in the size or the position of the cyst. Surgical treatment was performed two days after the delivery. There were no complications and no recurrences have occurred to date. PMID- 18412795 TI - Diagnosis of ovarian torsion by three-dimensional power Doppler in first trimester of pregnancy. AB - Three-dimensional ultrasonography in power Doppler mode (3D power Doppler) allows the appropriate quantitative assessment of the ovarian tissue vascularization. The present case is of an ovarian torsion in a pregnant woman at 6.5 weeks with ovarian hyperstimulation syndrome due to in vitro fertilization. Gray-scale images of endovaginal two-dimensional ultrasonography revealed enlargement of the right ovary and multiple peripheral follicles. Two-dimensional ultrasonography in color Doppler mode revealed the absence of venous flow and reduction in arterial flow to the impaired ovary. 3D power Doppler allowed the appropriate quantitative assessment of tissue vessel density of the impaired ovary, which greatly influenced the decision for conservative surgical treatment. The 3D power Doppler assessment performed after the surgery confirmed the integrity of the ovarian tissue. 3D power Doppler can help in the selection of patients with ovarian torsion for conservative surgical treatment. PMID- 18412796 TI - Mucoepidermoid carcinoma of the parotid gland with ovarian and peritoneal metastasis. AB - Metastatic ovarian carcinoma generally arises from the gastrointestinal tract. Mucoepidermoid carcinoma is the most common malignant tumor of the salivary gland. It typically disseminates to the regional lymph nodes and surrounding soft tissues. Distant metastasis of the parotid mucoepidermoid carcinoma is very rare, and it usually occurs to the lungs, bones and liver. A 28-year-old woman with a history of high grade mucoepidermoid carcinoma of the parotid gland was admitted with ascites and bilateral ovarian masses. Breast examination, endoscopic evaluation of the gastrointestinal tract, total bone scintigraphy, abdominopelvic and chest tomography and cytological analysis of the ascites did not detect a primary malignancy. Nevertheless, pathological examination of the ovaries revealed a metastasis from the mucoepidermoid carcinoma. Metastatic ovarian tumors usually originate from the gastrointestinal tract. Other sources of tumor metastasis should also be considered. PMID- 18412797 TI - Uterine choriocarcinoma accompanied by an extremely high human chorionic gonadotropin level and thyrotoxicosis. AB - The conventional treatments given to a 24-year-old woman with metastatic uterine choriocarcinoma and clinical and biochemical thyrotoxicosis did not appear to have any effect, probably due to an extremely high serum human chorionic gonadotropin (hCG) level which was up to 11,910,000 mIU/mL, and were initially underscored in light of the 'high-dose hook effect'. To our knowledge, no extremely high hCG level in a uterine choriocarcinoma patient has been reported in the literature. Her decapacitating symptoms subsided after the first course of chemotherapy by etoposide, methotrexate, and actinomycin D-cyclophosphamide and vincristine (EMA-CO) regimen. The serum hCG level, which reflects the quantification of host tumor burden, returned to the reference range after the fifth course of chemotherapy and the thyroid function reached euthyroid status before the third course of chemotherapy; two final courses were administered after the hCG level became undetectable. Two years after remission of disease, the patient experienced a normal pregnancy, and a term baby girl was delivered vaginally. No recurrence of uterine choriocarcinoma has been noted for 7 years. PMID- 18412798 TI - Endometrioid endometrial adenocarcinoma recurring as carcinosarcoma. AB - Mullerian carcinosarcoma is currently regarded as a metaplastic (sarcomatous) carcinoma. Only five cases of pure ovarian adenocarcinoma recurring as carcinosarcoma have been documented in the literature. There are no documented cases of endometrial adenocarcinoma recurring as metaplastic carcinoma. We report of a case of endometrial adenocarcinoma, endometrioid type, recurring as metaplastic carcinoma showing sarcomatous differentiation. The tumor evolution in this case supports the prevailing opinion that Mullerian carcinosarcomas are derived from carcinomas and represent tumor progression. PMID- 18412799 TI - Ovarian stromal luteoma in a patient with endometrial carcinoma. AB - A 65-year-old patient with endometrial adenocarcinoma underwent hysterectomy. Pathological examination of the right ovary revealed a coexisting stromal luteoma. The possible relationship between the two neoplasms as well as the intriguing pathogenesis and controversial nomenclature of ovarian steroid cell tumors are discussed. The incidental discovery of a stromal luteoma in a total hysterectomy specimen removed for endometrial carcinoma raises interesting questions about the perceived innocuous nature of these rare lipid cell tumors. PMID- 18412800 TI - Granulocyte-colony stimulating factor for the treatment of ritodrine-induced neutropenia. AB - We report on three pregnant women with ritodrine-induced neutropenia who were successfully treated with granulocyte-colony stimulating factor (G-CSF). The neutropenia occurred after continuous intravenous infusion of ritodrine for preterm labor. Ritodrine was discontinued and G-CSF was administered. Neutrophil counts returned to normal an average of 4.3 days after the administration. No infectious morbidity or adverse side-effects occurred in the mothers or infants. G-CSF is one possible treatment in women with ritodrine-induced neutropenia. PMID- 18412801 TI - Partial and complete HELLP, does the difference matter? PMID- 18412802 TI - Abstracts of the 11th Asian Pacific Congress of Nephrology, incorporating the 15th Asian Colloquium in Nephrology and 2nd Asian Forum of CKD Initiative. May 5 8, 2008. Kuala Lumpur, Malaysia. PMID- 18412803 TI - Abstracts of the Australasian College of Dermatologists 41st Annual Scientific Meeting. May 18-21, 2008. Sydney, Australia. PMID- 18412804 TI - Filaggrin and the great epidermal barrier grief. AB - One of the principal functions of human skin is to form an effective mechanical barrier against the external environment. This involves the maturation and death of epidermal keratinocytes as well as the assembly of a complex network of differentially and spatially expressed proteins, glycoproteins and lipids into the keratinocyte cell membrane and surrounding extracellular space. In 2006, the key role of the granular cell layer protein filaggrin (filament-aggregating protein) in maintaining the skin barrier was determined with the identification of loss-of-function mutations in the profilaggrin gene (FLG). These mutations have been shown to be the cause of ichthyosis vulgaris and a major risk factor for the development of atopic dermatitis, asthma associated with atopic dermatitis as well as systemic allergies. Mutations in the FLG gene are extremely common, occurring in approximately 9% of individuals from European populations. The remarkable discovery of these widespread mutations is expected to have a major impact on the classification and management of many patients with ichthyosis and atopic disease. It is also hoped that the genetic discovery of FLG mutations will lead to the future development of more specific, non immunosuppressive treatments capable of restoring effective skin barrier function and alleviating or preventing disease in susceptible individuals. PMID- 18412805 TI - Effect and persistency of botulinum toxin iontophoresis in the treatment of palmar hyperhidrosis. AB - The purpose of this study was to investigate the efficacy and persistence of the hypohidrosis induced by iontophoresis of botulinum toxin type A in patients with palmar hyperhidrosis. Eight patients with severe palmar hyperhidrosis participated in this study. Iontophoresis of botulinum toxin was applied to the patients' dominant (right) hand and the other hand was the control treated with normal saline. Gravimetry and iodine-starch tests were performed to evaluate the rate of sweating. The evaluation sessions were at baseline, 2 and 4 days, 1, 2, 3, 4, 8 and 12 weeks after treatment. The patients were also asked to fill out the Dermatology Life Quality Index questionnaire at 2, 4, 8 and 12 weeks after treatment. The mean sweating rate significantly reduced at the first, second, third and fourth week and the second and third month (P < 0.05) after the treatment in the right hand, and the third and fourth week (P < 0.05) after treatment in the left hand. Iontophoresis of botulinum toxin rapidly induced a quick reduction of sweating rate for at least 3 months with no pain, muscle weakness or other side-effects. PMID- 18412807 TI - Degos' disease with delayed involvement of the gastrointestinal tract. AB - A 75-year-old woman presented with pink papules over the trunk and limbs. Over the next few weeks, the papules became umbilicated and then developed into porcelain-white lesions with a rose or violaceous rim. A biopsy of one of these lesions revealed superficial and deep dermal lymphohistiocytic inflammation, mainly perivascular with a central zone of infarction consistent with Degos' disease. Three years later, she presented with increasing abdominal pain, anaemia, raised inflammatory markers and an abdominal bruit. Angiography showed splanchnic vessel stenosis possibly secondary to vasculitis and she underwent successful balloon dilatation. Treatment with prednisolone was commenced. There has been no recurrence of lesions in the 12 months since the balloon dilatation. PMID- 18412806 TI - Prospective study of patch testing in patients with vulval pruritus. AB - Forty-three patients with the primary complaint of vulval pruritus were recruited to take part in this prospective patch-testing study. A detailed questionnaire was administered to each and patch testing to an extended battery of allergens was undertaken. This included the European standard series, preservatives, corticosteroids and a battery of common over-the-counter topical vulval treatments. Analysis of demographic data and prior treatments used, and various other parameters, were studied in the context of patch-test results. This prospective study reports a very high rate of contact sensitivity in patients presenting with vulval pruritus, with 81.4% of those tested having at least one contact allergen detected. One or more clinically relevant allergens were found in 44% of the subjects patch tested. The variables found to predict a greater likelihood of a contact allergy were a biopsy diagnosis of vulval dermatitis, the use of multiple topical treatments, sexual inactivity and patients with severe pruritus on a self-reported scale. Most importantly, many clinically relevant allergens these patients reacted to were outside the European standard series. This highlights the need for an extended series in this patient population. PMID- 18412808 TI - Bullous systemic lupus erythematosus responding to dapsone. AB - A 29-year-old woman with a 4-week history of systemic lupus erythematosus presented acutely with a severe generalized tense vesicular and bullous eruption with involvement of mucosal surfaces. At the time of her initial diagnosis of systemic lupus erythematosus, she had declined treatment, preferring to explore complementary medical therapies. Skin biopsy showed subepidermal blister formation with inflammation at the dermoepidermal junction. Direct immunofluorescence revealed strongly positive linear deposition of IgG and IgM, and positive linear granular deposition of IgA along the basement membrane zone. Electron microscopy showed that the level of the basement membrane split was below the lamina densa. A diagnosis of bullous systemic lupus erythematosus was made and dapsone was commenced, with a dramatic improvement in her skin eruption. The patient again declined further treatment of her systemic disease and sought complementary therapies, and subsequently presented with cerebral involvement. PMID- 18412809 TI - Painful digital infarction in a male smoker treated as Buerger's disease. AB - A 40-year-old male smoker presented with a 3-week history of painful digital infarction involving the hands. Histology was unhelpful, showing lichen simplex chronicus. A provisional diagnosis of a vasculitic disease was made. Treatment included aspirin, azathioprine, iloprost, methylprednisolone, nifedipine and prednisolone. After failure of these treatments an alternative clinical diagnosis of Buerger's disease was made. Treatment was to advise the patient of the importance of continuing to abstain from smoking, to perform bilateral video assisted thoracoscopic sympathectomies and to commence folate supplementation. This led to marked improvement of his symptoms and healing of the digital infarction. PMID- 18412810 TI - Cutaneous chancroid in a visitor from Vanuatu. AB - A 23-year-old woman from Vanuatu presented to an Australian hospital with a 3 week history of a non-healing ulcer on the lower leg. A swab was submitted for a multiplex polymerase chain reaction designed to investigate genital ulcerative conditions. Haemophilus ducreyi was detected and the gene product was subsequently sequenced, confirming the diagnosis of cutaneous chancroid. The lesion responded to intramuscular benzathine penicillin. This report adds further evidence that cutaneous chancroid should be considered in the evaluation of skin ulcers in the south Pacific. PMID- 18412811 TI - Late-onset Epstein-Barr virus-negative post-transplant lymphoproliferative disorder presenting as ulcerated necrotic papules and nodules in a renal transplant patient. AB - We report a case of B-cell post-transplant lymphoproliferative disorder in a 57 year-old female 19 years postrenal transplant patient who presented with multiple, progressive, painful ulcerated necrotic papules and nodules over the left leg. Histopathological examination showed diffuse infiltration of the dermis by large atypical B-lymphoid cells, with a negative in situ hybridization test for Epstein-Barr virus. Gastrointestinal involvement was evident by the presence of atypical lymphoid cells in the peritoneal fluid. She only had partial response to localized radiotherapy and intravenous rituximab and died 4 months later of septicaemia. Unusual features highlighted in this case include the very late onset of disease 19 years post-transplant, Epstein-Barr virus negativity and the aggressive course of disease that did not respond to the reduction of immunosuppression, localized electron beam therapy and intravenous rituximab. PMID- 18412812 TI - Erythroplasia of Queyrat of the glans penis on a background of Zoon's plasma cell balanitis. AB - Erythroplasia of Queyrat of the glans penis developed in a 79-year-old uncircumcised man on a background of biopsy proven Zoon's plasma cell balanitis affecting the same site on the glans. The Zoon's plasma cell balanitis had been treated with topical pimecrolimus for 1 month prior to the development of clinically evident erythroplasia of Queyrat. He was subsequently treated with topical 5-fluorouracil 5% for 2 weeks, which resulted in clinical clearance. He has since been circumcised. PMID- 18412813 TI - Plaque-like dermatofibroma: A distinct and rare benign neoplasm? AB - Unusual large dermatofibromata are reported in a 40-year-old man and a 48-year old man, who both presented with plaques on a lower limb. The largest plaque in each case was well-defined, reddish brown, indurated and measured 50 mm x 30 mm and 70 mm x 40 mm, respectively. Several satellite lesions were present around the large plaques. Dermoscopic examination showed diffuse homogenous pigmentation in the absence of other diagnostic criteria for dermatofibroma. Light microscopy of biopsies from each patient displayed similar features. There was a dermal proliferation of fibrohistiocytic cells that entrapped intervening thickened collagen fibres. The overlying epidermis was acanthotic, and in some instances this showed basal hyperpigmentation. There was no evidence of malignancy. Immunohistochemical staining was positive for Factor XIIIa and negative for CD34. Based on the histological findings, a diagnosis of dermatofibroma was made for each of these cases. Fewer than 20 adult cases of large dermatofibroma of this scale, designated giant dermatofibroma, have been reported to date; and only two have shown a plaque-like appearance, the remainder being pedunculated. The authors propose plaque-like dermatofibroma as a variety of large dermatofibroma distinct to pedunculated giant dermatofibroma. PMID- 18412814 TI - Acitretin for lichen amyloidosus. AB - We present two cases of lichen amyloidosus treated with retinoids. A 57-year-old Vietnamese woman has had extensive generalized recalcitrant lichen amyloidosus for 23 years. Treatment with oral etretinate (25 mg/day) for 3 years, and later oral acitretin (10 mg/day) for the past 10 years, has controlled the pruritus and flattened the hyperkeratotic papules. Whenever the acitretin was ceased her symptoms flared within weeks. On each occasion reintroduction of acitretin was effective within 1-2 months. The second case is that of a 51-year-old Australian Aboriginal woman who had a 2-year history of lichen amyloidosus affecting her lower legs. A 2-month course of oral acitretin (25 mg b.d.) produced a marked improvement in both the pruritus and hyperkeratotic papules. She was then lost to follow up for 2 years, during which time her symptoms recurred. PMID- 18412815 TI - A case of sexually induced episodic urticaria. AB - A 33-year-old male experienced three episodes of urticaria precipitated by exposure to a hidden source of ampicillin. Each occurrence coincided temporally with his girlfriend taking ampicillin for a chest cold and/or allergies. We postulate that he was exposed to ampicillin in the body fluids of his sexual partner. PMID- 18412816 TI - Hypersensitivity reaction to the ingestion of mango flesh. AB - A 42-year-old woman presented with a hypersensitivity reaction after the ingestion of a small amount of fresh mango gelato. She developed itchy palpable purpuric lesions over her arms, legs, neck and abdomen 4 days after ingestion. The lesions persisted for 5 weeks despite treatment with betamethasone-17 valerate 0.05% ointment and avoidance of mango. Resolution of these lesions was eventually achieved with continuing treatment. The patient denied any prior contact with mango skin but had experienced previous sensitizing reactions to mango flesh. Patch testing was strongly positive to mango skin and mango flesh. Skin-prick testing was negative. This case describes a systemic contact dermatitis to mango flesh, an entity less common than allergic contact dermatitis. PMID- 18412821 TI - JMFT special section on mass trauma. PMID- 18412823 TI - Evaluating a multiple-family group access intervention for refugees with PTSD. AB - The purpose of this study was to analyze the effects of a multiple-family group in increasing access to mental health services for refugees with posttraumatic stress disorder (PTSD). This study investigated a nine-session multiple-family group called Coffee and Families Education and Support with refugee families from Bosnia-Herzegovina in Chicago. Adults with PTSD (n = 197) and their families were randomly assigned to receive either the intervention or a control condition. The results indicated that a multiple-family group was effective in increasing access to mental health services and that depression and family comfort with discussing trauma mediated the intervention effect. Further well-designed studies of family interventions are needed for developing evidence-based interventions for refugee families. PMID- 18412822 TI - When the levee breaks: treating adolescents and families in the aftermath of hurricane katrina. AB - Hurricane Katrina brought to the surface serious questions about the capacity of the public health system to respond to community-wide disaster. The storm and its aftermath severed developmentally protective family and community ties; thus its consequences are expected to be particularly acute for vulnerable adolescents. Research confirms that teens are at risk for a range of negative outcomes under conditions of life stress and family disorganization. Specifically, the multiple interacting risk factors for substance abuse in adolescence may be compounded when families and communities have experienced a major trauma. Further, existing service structures and treatments for working with young disaster victims may not address their risk for co-occurring substance abuse and traumatic stress reactions because they tend to be individually or peer group focused, and fail to consider the multi-systemic aspects of disaster recovery. This article proposes an innovative family-based intervention for young disaster victims, based on an empirically supported model for adolescent substance abuse, Multidimensional Family Therapy (MDFT; Liddle, 2002). Outcomes and mechanisms of the model's effects are being investigated in a randomized clinical trial with clinically referred substance-abusing teens in a New Orleans area community impacted by Hurricane Katrina. PMID- 18412824 TI - Beyond individual war trauma: domestic violence against children in Afghanistan and Sri Lanka. AB - To date, research on the psychosocial consequences of mass trauma resulting from war and organized violence on children has primarily focused on the individual as the unit of treatment and analysis with particular focus on mental disorders caused by traumatic stress. This body of research has stimulated the development of promising individual-level treatment approaches for addressing psychological trauma. In contrast, there is virtually no literature addressing the effects of mass trauma on the family and community systems. Research conducted in Sri Lanka and Afghanistan, two long-standing war-torn societies, found that in addition to multiple exposure to war or disaster-related traumatic events children also indicated high levels of exposure to family violence. These findings point to the need for conjoint family- and community-based programs of prevention and intervention that are specifically tailored for the context of the affected society. In particular, programs should take issues such as poverty, child labor, and parental alcohol use into account in assessing and treating children in the aftermath of mass trauma. PMID- 18412825 TI - Parenting practices as potential mechanisms for child adjustment following mass trauma. AB - Trauma research has identified a link between parental adjustment and children's functioning and the sometimes ensuing intergenerational impact of traumatic events. The effects of traumatic events on children have been demonstrated to be mediated through their impact on children's parents. However, until now, little consideration has been given to the separate and more proximal mechanism of parenting practices as potential mediators between children's adjustment and traumatic events. To shed some light in this arena, we review literature on trauma, adversity, and resilience, and discuss how parenting practices may mediate trauma and adverse environmental contexts. Using a social interaction learning perspective (Forgatch & Knutson, 2002; Patterson, 2005), we propose a prevention research framework to examine the role that parenting practices may play in influencing children's adjustment in the wake of trauma exposure. The article concludes by providing a specific model and role for evidence-based parenting interventions for children exposed to mass trauma. PMID- 18412826 TI - Linking human systems: strengthening individuals, families, and communities in the wake of mass trauma. AB - This article presents an overview of the philosophy and practical principles underlying the Linking Human Systems Approach based on the theory of resilience in individuals, families, and communities facing crisis, trauma, and disaster. The Link Approach focuses on tapping into the inherent strength of individuals and their families and emphasizes resilience rather than vulnerability. It has been successfully used in combating critical public health problems, such as addiction, HIV/AIDS, and recovery from major trauma or disaster. Also, three specific models of Link intervention aimed at the individual, family, and community levels are discussed, with special emphasis on the family-level intervention. These interventions are directed toward mobilizing resources for long-term physical, emotional, psychological, and spiritual healing. PMID- 18412827 TI - A Virginia Tech MFT ethics class reflects on the shootings at Virginia Tech. AB - The authors of this article include the professor and most of the students in a doctoral course on marriage and family therapy ethical and professional issues that met the semester that a disturbed student shot and killed 32 Virginia Tech students and faculty before killing himself. In this article, we reflect through short essays on issues related to the tragedy, ethics, and recovery. PMID- 18412828 TI - A systematic review of the research base on sexual reorientation therapies. AB - In the past few years, members of the AAMFT, like members of other professional groups, have engaged in a discourse as to the necessity and effectiveness of sexual reorientation therapies. The purpose of this article is to review, critique, and synthesize the scientific rigor of the literature base underpinning sexual reorientation therapy research. Using a systematic narrative analysis approach, 28 empirically based, peer-reviewed articles meeting eligibility criteria were coded for sample characteristics and demographics as well as numerous methodology descriptors. Results indicate the literature base is full of omissions which threaten the validity of interpreting available data. PMID- 18412829 TI - Family therapist comfort with and willingness to discuss client sexuality. AB - Limited empirical information exists on whether or not marriage and family therapists are having sexuality-related discussions with their clients. When helping professionals ignore client sexuality, the potential for unintended negative outcomes increases. The researchers surveyed 175 clinical members of the American Association for Marriage and Family Therapy to assess how their clinical training and education, their perceived sexual knowledge, and their comfort with sexual material influenced their willingness to engage in sexuality-related discussions with their clients. The results indicate that sexuality education and supervision experiences are the cornerstone for a therapist's base level of comfort. It is through sexuality education and supervision that sex knowledge is acquired and comfort levels are increased. Once comfort with sexual discussions increases, then therapists are more likely to engage in sexuality discussions with their clients. PMID- 18412830 TI - Relational safety and liberating training spaces: an application with a focus on sexual orientation issues. AB - This article describes and discusses a teaching case of a clinical training situation involving a gay marriage and family therapy student working with a same sex affectional couple. The conceptual pillars of this teaching case, relational safety and liberating spaces, are advanced as illustrations of how the student developed his voice in the training process. Pivotal moments in this process are discussed, as are implications for training and personal and professional growth. PMID- 18412833 TI - Consultation frequency and perceived consultation time in a Japanese psychiatric clinic: their relationship with patient consultation satisfaction and depression and anxiety. AB - AIM: To examine the relationship of the consultation frequency and the perceived consultation time of psychiatric clinic attenders with their satisfaction towards the consultation and depression and anxiety (measured by the Hospital Anxiety and Depression Scale). METHODS: Questionnaires were distributed to 186 outpatients attending a psychiatric clinic. RESULTS: In a path analytic model, a longer session with shorter interval ('dense' session) predicted lowered depression and anxiety level of the patientsthrough the perceived satisfaction of the patients, and it was also predicted by the higher anxiety level. The dense session was also influenced by shorter duration of clinic attendance. CONCLUSION: Dense psychiatric sessions in busy Japanese outpatient clinics may benefit patients. PMID- 18412834 TI - Measurement of development of cognitive and attention functions in children using continuous performance test. AB - AIM: The development of attention function in children is still not sufficiently clear. Although it is difficult to objectively assess attention function, continuous performance tests (CPT) can be used to objectively assess cognitive function along with attention. The development of cognitive and attention functions was examined in children using a CPT. METHODS: A total of 541 healthy girls aged 5-12 years participated. Ten parameters were calculated: numbers of cancellations for either target stimuli (T-cancel) or non-target stimuli (N cancel), numbers of omission errors (Omission) and commission errors (Commission), hit rate (Hit), false alarm rate (False), mean reaction time for correct response (RT), coefficient of variance for mean reaction time (CVRT), sensitivity index (d'), and lnbeta. RESULTS: The parameters were divided into three types based on pattern of change. T-cancel, False, and Commission, which are related to inhibition of response, N-cancel, Hit, and Omission, which are related to inattention to stimuli, and CVRT, which is related to stability of processing time, exhibited significant change until 5 or 6 years of age. d', which is related to ability to discriminate between target or non-target, exhibited significant change until 8 years of age. RT, which is related to processing time, exhibited significant change until 11 years of age. lnbeta exhibited no significant differences among age groups. CONCLUSIONS: These findings indicate that inhibition function, inattention to stimuli, and stability of processing time develop first. Discrimination ability subsequently increases based on these developments, and finally processing time is reduced. PMID- 18412835 TI - Factor structure of the Japanese Interpersonal Competence Scale. AB - AIM: Assessing social competence is important for clinical and preventive interventions of depression. The aim of the present paper was to examine the factor structure of the Japanese Interpersonal Competence Scale (JICS). METHODS: Exploratory and confirmatory factor analysis was performed on the survey responses of 730 participants. Simultaneous multigroup analyses were conducted to confirm factor stability across psychological health status and sex differences. RESULTS: Two factors, which represent Perceptive Ability and Self-Restraint, were confirmed to show a moderate correlation. Perceptive Ability involves a more cognitive aspect of social competence, while Self-Restraint involves a more behavioral aspect, both of which are considered to reflect the emotion-based relating style specific to the Japanese people: indulgent dependence (amae) and harmony (wa). In addition, Self-Restraint may be linked to social functioning. Both constructs may confound a respondent's perceived confidence. CONCLUSION: Despite its shortcomings, the JICS is a unique measure of social competence in the Japanese cultural context. PMID- 18412836 TI - Reevaluating the incidence of pervasive developmental disorders: impact of elevated rates of detection through implementation of an integrated system of screening in Toyota, Japan. AB - AIM: Although recent epidemiological studies on the pervasive development disorders (PDD) appear to be reporting higher rates of incidence than previously believed, great variation in the reported figures suggests a need for review of the methodology involved. As such, a survey on the incidence of PDD was conducted and compared with data from a previous survey to examine the effects of screening and diagnostic methodology on incidence. METHODS: The incidence of pervasive developmental disorders was surveyed in all children (12 589) born between January 1994 and December 1996 in Toyota, Japan. RESULTS: Incidence was 1.81% and the ratio of boys to girls was 2.80. Definitive diagnoses were made between 13 months and 7 years 2 months, the average age at diagnosis being 3 years 4 months. Among the cases of PDD, children with normal or borderline intelligence amounted to 66.4%, mild mental retardation (MR) 17.5%, moderate MR 10.3% and severe MR 5.8%. CONCLUSION: An approximately 11-fold increase was noted in prevalence of PDD compared to a previous survey two decades ago, and two main factors were believed to account for this apparent sharp increase. First, inclusion of high functioning subjects detected during infancy, and second, higher rates of diagnosis resulting from an integrated process of screening. PMID- 18412837 TI - Patterns of temperament and character in a clinical sample of Korean children with attention-deficit hyperactivity disorder. AB - AIMS: The purpose of the present study was to assess patterns of temperament and character in a clinical sample of Korean children with attention-deficit hyperactivity disorder (ADHD) and to investigate the relationship between patterns of temperament and character and the severity of ADHD symptoms. METHODS: Fifty-one children who met DSM-IV criteria for ADHD and 51 age- and sex-matched healthy comparison subjects completed the Junior Temperament and Character Inventory (JTCI) and the DuPaul's ADHD Rating Scale (ARS-IV). RESULTS: Children with ADHD had significantly higher scores of Novelty Seeking and lower scores of Self-directedness in both parents' rating and children's self-rating. In addition, low Self-directedness scores predicted higher total scores of ARS-IV. CONCLUSIONS: In a clinical sample of Korean children with ADHD, a distinct pattern of temperament and character was found. The current findings suggest that low Self-directedness may be related to the severity of ADHD symptoms. PMID- 18412838 TI - Poor performance in Clock-Drawing Test associated with visual memory deficit and reduced bilateral hippocampal and left temporoparietal regional blood flows in Alzheimer's disease patients. AB - AIM: To investigate the associations of Clock-Drawing Test (CDT) score with neuropsychological test scores and regional cerebral blood flow. METHOD: Twenty five patients (normal aging, n = 2; mild cognitive impairment, n = 7; Alzheimer's disease [AD], n = 16) participated in this study. Their average age was 74.8 years. RESULTS: CDT score correlated well with the neuropsychological test scores of Mini-Mental State Examination, Clinical Dementia Rating, immediate visual memory, delayed visual memory, and IQ evaluated by Koh's block design. CDT score also had a statistically significant correlation with the regional blood flow in the left hippocampal region as evaluated on 3-D stereotaxic region-of-interest template analysisapplied to single-photon emission computed tomography images. Using a cut-off point of 8/9 in the CDT, the high-CDT group had significantly higher delayed visual memory and IQ scores than the low-CDT group. Moreover, the high-CDT group had significantly higher regional blood flows in the left parietal, left angular and bilateral hippocampal regions than the low-CDT group. CONCLUSION: CDT score correlates well with regional cerebral blood flow that is decreased in the early stage of AD. PMID- 18412839 TI - Relationships among burnout, coping style and personality: study of Japanese professional caregivers for elderly. AB - AIM: To investigate relationships among burnout, coping style and personality. METHODS: Seventy-two professional caregivers working at nursing homes in Japan were assessed using the Maslach Burnout Inventory (MBI), NEO Five-Factor Inventory (NEO-FFI), 30-item General Health Questionnaire (GHQ) and Coping Inventory for Stressful Situation (CISS). RESULTS: GHQ (beta = 0.34, P < 0.01) and emotion-oriented coping (CISS-E; beta = 0.31, P < 0.05) were the predictors of emotional exhaustion (MBI-EE), and neuroticism (NEO-N; beta = 0.45, P < 0.001) and age (beta = -0.23, P < 0.05) were the predictors of depersonalization (MBI DP). Structural equation modeling showed that NEO-N affected the burnout scores through CISS-E, and that age and GHQ affected the burnout scores independently. CONCLUSION: Personality affects burnout through coping style in professional caregivers. PMID- 18412840 TI - Alterations in prefrontal cortical activity in the course of treatment for late life depression as assessed on near-infrared spectroscopy. AB - AIM: To evaluate the severity of depression by measuring alterations in prefrontal cortical activity associated with mood disorders, as assessed on near infrared spectroscopy. METHODS: Ten of 27 subjects hospitalized for late-life depression from May 2006 to June 2007, were examined. In these 10 subjects changes in hemoglobin concentration were measured on near-infrared spectroscopy during two types of the rock, paper, scissors game as the cognitive tasks affecting prefrontal cortical activity on 2 days, including 1 day on which depressive symptoms had slightly improved due to treatment, and then on another day >4 weeks later. Severity of depression and cognitive impairment were also simultaneously assessed. RESULTS: The change in oxygenated hemoglobin concentration during a difficult task (intentional loss task) was significantly larger than that during an easy task (try to win task) on the left side (left, P = 0.010; right, P = 0.059). On the left side there was a significant negative correlation between the ratio of oxygenated hemoglobin measurements on the second day to those on the first day, and the severity of depression on the second day (left, P = 0.012; right, P = 0.090). Thus, the more left prefrontal cortical activity tended to increase, the fewer depressive symptoms tended to be present on the second day of testing. CONCLUSIONS: Measuring of alterations in prefrontal cortical activity associated with mood disorders, as assessed on near-infrared spectroscopy, is feasible in subjects with depression. PMID- 18412841 TI - Effect of mood states on QT interval and QT dispersion in eating disorder patients. AB - AIM: Prolonged QT interval and QT dispersion have been reported in patients with eating disorders. Although the factors that cause prolongation remain unclear, mood states such as anxiety have been reported to influence QT interval and dispersion, probably via the autonomic nervous system. Therefore the aim of the present paper was to investigate mood effect on prolonged QT interval and QT dispersion. METHOD: The subjects were 47 female anorexia nervosa (AN) and 48 female bulimia nervosa (BN) patients. In all of the patients, serum electrolyte levels were normal. QT interval and QT dispersion were measured from 12-lead electrocardiographic recordings. Mood states in each patient were measured using a Profile of Mood States (POMS) evaluation, and the patients were divided into high- and low-score groups for each POMS subscale. The differences in QT variables were compared between the two groups for each subscale. RESULTS: In the BN group, QT interval and QT dispersion in the high depression score group were significantly longer than those in the low depression score group, and QT dispersion was significantly greater in the high anxiety score group than in the low anxiety score group. In addition, QT interval and QT dispersion were significantly correlated with depression scores. In the AN group there were no significant differences in QT interval or QT dispersion between the high- and low score groups for any POMS subscale. CONCLUSIONS: BN patients with worse states of depression or anxiety had longer QT intervals and larger QT dispersion. In BN patients, mood disturbance might increase the risk of arrhythmias. PMID- 18412842 TI - Neurocognition in first-degree healthy relatives (siblings) of bipolar affective disorder patients. AB - AIM: Cognitive deficits have been presupposed to be endophenotypic markers in bipolar disorder, but few studies have ascertained the cognitive deficits in healthy relatives of bipolar disorder patients. The aim of the present study was to assess the cognitive functions of first-degree relatives of patients with bipolar disorder and compare them with healthy controls. METHODS: Ten first degree apparently healthy relatives of patients with bipolar disorder were compared with 10 age- and education-matched control subjects on computer-based cognitive tests. RESULTS: As compared to the control group, the relatives group performed significantly poorly on tests for executive function and vigilance, while on the test for working memory the performance was not significantly different on most of the parameters. CONCLUSIONS: Executive functioning and vigilance could be potential markers of the endophenotype in bipolar patients. PMID- 18412843 TI - No predictors of antidepressant patient response to milnacipran were obtained using the three-factor structures of the Montgomery and Asberg Depression Rating Scale in Japanese patients with major depressive disorders. AB - AIMS: Milnacipran, a new specific serotonin and norepinephrine re-uptake inhibitor, is as effective as tricyclic antidepressants. Symptomatological predictors of antidepressant response to milnacipran have not been studied until now. METHODS: This study included 101 Japanese patients who fulfilled the DSM-IV criteria for the diagnosis of major depressive disorders and whose score on the Montgomery and Asberg Depression Rating Scale (MADRS) was > or =21. Eighty-three patients were finally included. Patients with a pretreatment MADRS score > or =31 points were defined as severe (n = 28), and the rest as non-severe (n = 55). The three-factor model of MADRS was used for analysis; the first factor was defined by three items, the second factor was defined by four items and the third factor was defined by three items representing dysphoria, retardation, and vegetative symptoms, respectively. Milnacipran was administered twice daily for 6 weeks. The initial dose was 50 mg/day; after a week it was increased to 100 mg/day. RESULTS: No significant difference was observed in the mean score of first factor, second factor and third factor at pretreatment time between responders and non responders in both severe and non-severe patients. CONCLUSIONS: No predictor of antidepressant response to milnacipran was obtained using the three-factor structures of the MADRS in Japanese patients with major depressive disorders. PMID- 18412844 TI - Reward pathways in Parkinson's disease: clinical and theoretical implications. AB - AIMS: The mesolimbic and mesocortical circuits are particularly involved in reward-related behavior in humans. Because these systems may be in some way altered in Parkinson's disease (PD), it is likely that some psychiatric manifestations of PD, such as hedonistic homeostatic dysregulation and pathological gambling, as well as impulsive decision making, may be ascribed to their involvement. The aim of the current article was to review recent literature on this topic in order to analyze whether these disturbances share a common ground and whether actual theoretical frameworks on addiction prove a useful tool for their interpretation. METHODS: Data were identified on searches of MEDLINE/PubMed databases from relevant articles published up until March 2007. RESULTS: All clinical manifestations (hedonistic homeostatic dysregulation, pathological gambling and impulsive decision making) seem to share a common multifaceted ground in which factors related to antiparkinsonian therapy, premorbid personality and progression of disease interact. Theoretical interpretations and conclusions drawn from experimental studies may help to shed light on the underlying pathological mechanisms. CONCLUSIONS: Further studies are needed to analyze why, despite a common ground, only some patients develop those neuropsychiatric complications described here. PMID- 18412845 TI - Enhanced activation in the extrastriate body area by goal-directed actions. AB - AIM: Neuroimaging studies on biological motion have established the view that the posterior superior temporal sulcus (pSTS) is involved in detecting intention of others. Those studies have consistently reported other regions such as body selective extrastriate body area (EBA) and motion-sensitive middle temporal, in close proximity to pSTS. Whether EBA responds only to static body parts or has a more extended role as part of a system for inferring intention of others has remained an elusive issue. The aim of the present study was to investigate the role of EBA in processing goal-directed actions. METHODS: Twelve healthy volunteers participated in the present study. Using sports-related motions as visual stimuli, brain activations were examined during observation of goal directed actions and non-goal-directed actions on functional magnetic resonance imaging. RESULTS: Compared to non-goal-directed actions, goal-directed actions produced greater activations in EBA along with the mirror neuron system. CONCLUSIONS: EBA might contribute to understanding others' actions by representing the dynamic aspects of human motions. PMID- 18412846 TI - Confabulation behavior and false memories in Korsakoff's syndrome: role of source memory and executive functioning. AB - AIMS: Confabulation behavior is common in patients with Korsakoff's syndrome. A distinction can be made between spontaneous and provoked confabulations, which may have different underlying cognitive mechanisms. Provoked confabulations may be related to intrusions on memory tests, whereas spontaneous confabulations may be due to executive dysfunction or a source memory deficit. METHODS: In 19 chronic Korsakoff patients, spontaneous confabulations were quantified by third party rating (Likert scale). Provoked confabulations were assessed using the Dalla Barba Confabulation Battery. Furthermore, assessment of executive function was performed using an extensive neuropsychological battery. False memories (i.e. intrusions) and source memory were measured using twoparallelversions of a word list learning paradigm (a modification of the Rey Auditory Verbal Learning Test). RESULTS: There were deficits in source memory, in which patients incorrectly assigned previously learned words to an incorrect word list. Also, Korsakoff patients had extensive executive deficits, but no relationship between the severity of these deficits and the severity of confabulation or intrusions on a memory task was found. CONCLUSION: The present findings provide evidence for a dissociation between spontaneous confabulation, provoked confabulation and false memories. PMID- 18412847 TI - Reliability and validity of the Pervasive Developmental Disorders Assessment System. AB - AIM: To test the reliability and validity of the Pervasive Developmental Disorders Assessment System (PDDAS), a Japanese semistructured interview system. METHODS: The PDDAS, consisting of 91 items including 12 major items corresponding to 12 items in criterion A of DSM-IV autistic disorder criteria, 36 items on autistic symptoms and three Asperger's disorder (AS) screening items for diagnosing pervasive developmental disorders (PDD) and their subtypes and 40 items for other information including early development and past/family histories, was administered to mothers of 77 PDD children and 64 non-PDD children. RESULTS: The PDDAS had satisfactory interrater reliability (ranges of kappa, r and raw agreement rate were 0.69-1.00 in 76 items, 1.00 in 11 items and 0.91-1.00 in four kappa un-calculable items, respectively). Thirty-three of the 36 items and all of the 12 major items scored significantly higher in the PDD than non-PDD groups to show satisfactory discriminant validity. PDDAS and consensus DSM-IV diagnoses agreed in the 77 children in PDD diagnosis and disagreed in only two children in subtype diagnoses of autistic disorder and PDD not otherwise specified. CONCLUSIONS: The PDDAS, which takes 1.5 h to administer, seems to have clinical and research utility, although further investigation is necessary. PMID- 18412848 TI - Characteristics of fatigue in panic disorder patients. AB - It was suggested that fatigue is one of characteristics of panic disorder. Fatigue was assessed in 360 patients with panic disorder using the Japanese version of the Multidimensional Fatigue Inventory (MFI-J). The scores for general fatigue and reduced activity were significantly higher in the patients than in the controls. These tendencies were also observed in men when the subject group was differentiated according to sex, but not in women. In contrast, the trend for higher score for physical fatigue was observed only in the female patients. Thus, the present study suggests that the characteristics of fatigue vary with sex in panic disorder. PMID- 18412849 TI - Comorbid childhood gender identity disorder in a boy with Asperger syndrome. PMID- 18412850 TI - Large-scale case-control study of a functional polymorphism in the glutamate receptor, metabotropic 3 gene in patients with schizophrenia. PMID- 18412851 TI - Venlafaxine: Successful treatment in impulsive disorders. PMID- 18412852 TI - 2C-T-4 intoxication: acute psychosis caused by a designer drug. PMID- 18412853 TI - Decreased blood perfusion in right thalamus after transient global amnesia demonstrated by an automated program, 3DSRT. PMID- 18412854 TI - Pramipexole-induced psychosis in Parkinson's disease. PMID- 18412855 TI - Serotonin syndrome induced by augmentation of SSRI with methylphenidate. PMID- 18412856 TI - Orthogonal polarization technique in the assessment of human skin microcirculation. AB - BACKGROUND: The "gold standard" for the study of the in vivo microcirculation is intravital microscopy. The recently developed method of orthogonal polarization of light [orthogonal polarization spectral (OPS) imaging] allows for the in vivo transcutaneous evaluation of the microcirculation without the need for invasive surgical procedures. METHODS: The application of polarized light originating from a 100 W halogen tungsten lamp is able to penetrate tissues at a depth of up to 3 mm, and generates reissued light from this depth. The evaluation of this depolarized light, from a deeper origin, may be carried out separately from the light reflected by the more superficial layers of the tissue under study because this light retains photon polarization, whereas the former light undergoes real depolarization. RESULTS: The process of validation of the OPS technique, when compared with intravital microscopy, the "gold standard" for the in vivo observation of the microcirculation, has shown that it is as effective and reliable as the gold standard, reaching the same resolution level in the visualization of blood vessels, but without the need for invasive surgical procedures. CONCLUSIONS: The OPS technique is a very promising tool for dermatologists and researchers, especially in the study of vasculitis, chronic venous insufficiency, and skin tumors. PMID- 18412857 TI - Olmsted syndrome: a case report and review of literature. AB - We report a case of an 18-month-old boy with slightly whitened fingernails and toenails since birth. At the age of 100 days, he progressively developed bilateral palmoplantar keratoderma which resulted in painful walking and disabled grasping. Perianal keratotic plaques and perioral hyperkeratotic erythema could also be observed. Both fingernails and toenails were dystrophic. Scalp hairs were sparse, but total alopecia was no present. The histopathologic changes of the biopsy from the inner side of the right foot showed nonspecific changes, which mainly showed highly hyperkeratosis and acanthosis with slight superficial perivascular inflammatory infiltration. A clinical diagnosis of Olmsted syndrome was established according to the typical feature of the lesions, which is the presence of symmetrical palmoplantar keratoderma with periorificial keratotic plaques. We review the literature and present a summary of all reported cases to date. PMID- 18412858 TI - Immunohistochemical characterization of the cellular infiltrate in localized scleroderma. AB - BACKGROUND: Localized scleroderma is a connective tissue disorder with hardening of the skin and fibrosis of the affected tissue as the most prominent features. The etiology of localized scleroderma is still unknown, but immunologic factors may play an important role in the pathogenesis. This study was performed to determine the immunohistochemical features of the cellular infiltrate in localized scleroderma. METHODS: Skin samples were obtained from six patients by 6 mm punch biopsy. The samples were stained with monoclonal antibodies against CD1a, CD3, CD4, CD8, CD20, CD25, CD30, and CD57. The number of cells stained with each monoclonal antibody was calculated. RESULTS: There were more CD1a+, CD3+, CD4+, CD8+, CD20+, CD25+, and CD57+ cells in the dermal infiltrate in localized scleroderma relative to those in normal controls. The numbers of CD1a+, CD3+, CD4+, CD8+, and CD57+ cells in localized scleroderma were significantly greater than those in normal skin (P < 0.05). The number of CD30+ cells in localized scleroderma was almost the same as that in normal skin. CONCLUSIONS: This study reveals that T lymphocytes, Langerhans cells, and natural killer cells may play important roles in the pathogenesis of localized scleroderma. PMID- 18412859 TI - Connexin 26 mutation in keratitis-ichthyosis-deafness (KID) syndrome in mother and daughter with combined conductive and sensorineural hearing loss. AB - BACKGROUND: Keratitis-ichthyosis-deafness (KID) syndrome most commonly results from a mutation in the gap-junctional protein connexin 26 (Cx26) gene, GJB2. Most cases are sporadic and are associated with sensorineural hearing loss. METHODS: We encountered a mother and daughter with KID syndrome, and pursued genetic analysis and an extensive hearing loss evaluation. RESULTS: The analysis of genomic DNA of both affected patients revealed the mutation 148G --> A in GJB2 (D50N). No mutation was found in an unaffected son. Auditory phenotype analysis showed a combined conductive and sensorineural hearing loss in both affected patients. CONCLUSIONS: This is the second vertical transmission of the D50N mutation. These are the first two cases with combined sensorineural and conductive hearing loss without any significant history of middle ear disease. This points to the possibility that the Cx26 D50N mutation can cause conductive hearing loss. PMID- 18412860 TI - Matrine inhibits invasiveness and metastasis of human malignant melanoma cell line A375 in vitro. AB - BACKGROUND: Matrine is a traditional Chinese medicine with significant inhibitory activity against malignant tumors. Its effects on the invasiveness and metastasis of malignant tumors have rarely been reported. AIM: To investigate whether matrine can inhibit the metastasis-related activities of the human malignant melanoma cell line A375 in vitro. METHODS: 3-(4,5-Dimethylthiazol-2-yl)-2,5 diphenyl tetrazolium bromide (MTT) assay and Annexin-V-fluorescein isothiocyanate/propidium iodide (Annexin-V-FITC/PI) affinity assay were used to examine the effects of matrine on the proliferation and apoptosis induction of A375 cells. The morphologic changes of A375 cells were observed by light and electron microscopy. Semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting were performed to evaluate the expression of heparanase mRNA and protein. The effect of matrine on the adhesion ability and invasiveness of treated A375 cells was tested by cell-Matrigel adhesion assay and Matrigel invasion assay, respectively. RESULTS: Matrine showed significant inhibition of the proliferation of A375 cells in a dose- and time-dependent manner. It also induced apoptosis in a dose-dependent manner. Compared with the control group, the levels of heparanase mRNA and protein expression of A375 cells treated with different concentrations of matrine were decreased significantly, as were their adhesion ability and invasiveness. CONCLUSIONS: These findings indicate that matrine inhibits the invasiveness and metastasis of A375 cells in vitro. The mechanisms may be linked to the inhibition of cellular proliferation, induction of apoptosis, and downregulation of heparanase mRNA and protein expression. PMID- 18412861 TI - LL-37 regulates the overexpression of vascular endothelial growth factor (VEGF) and c-IAP-2 in human keratinocytes. AB - BACKGROUND: The antimicrobial peptide PR39 is a porcine cathelicidin with angiogenic and antiapoptotic activities, as it can regulate the expression of vascular endothelial growth factor (VEGF) and inhibitor apoptosis protein-2 (c IAP-2) in endothelial cells. The human homolog LL-37 has been found to be highly expressed in human keratinocytes from psoriatic patients, but it is not known whether LL-37 can modulate the expression of VEGF and c-IAP-2 in keratinocytes, as both molecules are involved in the overgrowth of psoriatic skin. Therefore, in this work, we studied the possible role of CAP18/LL-37 in the modulation of VEGF and c-IAP-2 expression in human keratinocytes. METHODS: The CAP18/LL-37 gene was cloned into a plasmid that contained green fluorescent protein (GFP). This plasmid was called pGFP-CAP18/LL-37. The expression of LL-37, VEGF, and c-IAP-2 was determined by reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blotting in HaCaT cells transfected with pGFP-CAP18/LL-37. Specific DNAzymes were used to break the CAP18/LL-37 mRNA (DNAz-CAP18/LL-37). RESULTS: HaCaT cells transfected with pGFP-CAP18/LL-37 showed the upregulation of VEGF and c-IAP-2 mRNAs. Hypoxia-inducible factor-1alpha (HIF-1alpha) mRNA expression did not change during the assays; however, its protein was increased, as well as the VEGF protein. HaCaT cells cotransfected with pGFP-CAP18/LL-37 and DNAz-CAP18/LL 37 showed depleted expression of LL-37, VEGF, and c-IAP-2 mRNAs. CONCLUSIONS: These results suggest that LL-37 may modulate the expression of VEGF and c-IAP-2 via HIF-1alpha in human keratinocytes. PMID- 18412862 TI - Skin problems in amputees: a descriptive study. AB - BACKGROUND: Skin problems are common in amputee patients. These problems may restrict the normal use of a prosthetic limb. We aimed to determine the range, incidence, causes and patterns of dermatological problems seen in a population of amputees. MATERIALS AND METHODS: One hundred and forty two amputees, were enrolled to the study. Age, sex, age at the time of amputation, level of amputation, reason for amputation, and types of prosthesis were noted. Dermatological problems were recorded. Stumps were swabbed for bacteriological and mycological examination, and patch tests were performed in suspected patients. RESULTS: Of these 142 patients, 139 (97.9%) were males and 3 (2.1%) were females. The reasons for amputation in the majority of the cases were wounds due to mine explosion (n = 114, 80.3%) and gunshot wounds (n = 19, 13.4%). The other reasons were arterial diseases, traffic accidents, congenital absence of the tibia, and vascular complication of diabetes. At least one skin problem was detected in 105 (73.9%) of 142 cases. Positive reactions to allergens have been detected in 28 (43%) of 65 cases with dermatitis. Bacterial infection was detected in 12 patients and fungal infection was detected in 4 patients. CONCLUSION: Our descriptive study shows that skin problems have a high prevalence, up to 73.9% in amputee patients. This high percentage indicates that dermatological problems are important in amputees. Early recognition and treatment of these problems can prevent the amputee's mental, social, and economic losses. PMID- 18412863 TI - Metabolic syndrome associated with toenail onychomycosis in Taiwanese with diabetes mellitus. AB - BACKGROUND: Onychomycosis is a complication of diabetes mellitus (DM), which has a deleterious impact on the quality of life. Aim To explore the prevalence of onychomycosis amongst Taiwanese diabetics, and to analyze the factors associated with onychomycosis after adjusting for age and sex. METHODS: A total of 1245 Taiwanese diabetics were enrolled, and a nested case-control study was performed by onychomycosis outcome and the exposures were compared. RESULTS: The overall prevalence of onychomycosis among DM patients was 30.76% (383/1245), with a significantly higher prevalence in men than in women (P = 0.024). The factors associated with onychomycosis in matched pairs by gender and age were analyzed in 375 pairs. It was found that metabolic syndrome, obesity, triglyceride (TG) levels, and glycosylated hemoglobin (HbA1c) were associated with onychomycosis (P < 0.05). CONCLUSION: Higher prevalence rates of onychomycosis were found in men and older DM patients. Metabolic syndrome, obesity, high TG levels, and poor glycemic control were associated with onychomycosis. PMID- 18412864 TI - Dermatitis cruris pustulosa et atrophicans--a frequent but poorly understood tropical skin condition--a case report from Burkina Faso. AB - Dermatitis cruris pustulosa et atrophicans (DCPA) is a benign inflammatory skin disease of the younger population in the tropics. Although this pustular skin condition of particular topography is frequently seen by dermatologists in tropical countries, its origin remains unknown. We report the case of a young woman with DCPA associated with prurigo nodularis. A bacterial origin has not been demonstrated in this case. Histology showed an intraepidermal neutrophilic pustule with dermal and subcutaneous infiltration by neutrophils and eosinophils forming flame figures. Different pathogenic hypotheses are discussed with special regard to a potential relationship between DCPA and eosinophilic cellulitis. PMID- 18412865 TI - Multiple facial seborrheic keratosis-like lesions in a young woman with epidermodysplasia verruciformis. PMID- 18412866 TI - Tuberculous primary complex of the skin. PMID- 18412867 TI - Terra firma-forme dermatosis. PMID- 18412868 TI - Tinea pedis in athletes. PMID- 18412869 TI - A case of Degos disease with pleuropericardial fibrosis, jejunal perforation, hemiparesis, and widespread cutaneous lesions. PMID- 18412870 TI - Primary cutaneous carcinosarcoma: case report with expanded immunohistochemical analysis. PMID- 18412871 TI - Cutaneous pseudolymphoma associated with molluscum contagiosum: a case report. PMID- 18412872 TI - Type B pigmentary demarcation lines in a Chinese amenorrheic woman. PMID- 18412873 TI - A randomized investigator-blind trial of different passes of microdermabrasion therapy and their effects on skin biophysical characteristics. AB - BACKGROUND: Microdermabrasion (MDA) is a safe, simple, and beneficial technique for superficial skin resurfacing. Despite its popular usage, few studies have assessed the efficacy of different MDA protocols applied at the present time. Objectives To assess the effects of MDA generally, as well as to compare the effects of two vs. three passes of MDA in each session for a total number of six therapeutic sessions on skin biophysical characteristics. METHODS: In this randomized, investigator-blind, split-face study, 10 patients underwent a series of six MDA treatments with an interval of 2 weeks. One side of the face was treated with two passes of MDA and the other side was treated with three passes, randomly. Stratum corneum hydration, sebum secretion, and skin pH measurements were obtained before and after the procedure on all sessions and also 1 and 4 weeks after the last treatment. RESULTS: After six sessions of MDA, a decrease in sebum content compared to baseline was shown at the end of treatment sessions, but no statistical difference was observed between two vs. three passes groups ( 30.0 [interquartile range, IQR = 50.0] vs. -27.5 [IQR = 125.3], respectively, P = 0.58). Comparison of two treatment groups showed significant higher values of sebum content in the first follow-up after treatment with three passes of MDA. (64.0 [IQR = 52.0] for three passes vs. 45.0 [IQR = 46.0] for two passes, P = 0.04) A significant increase was observed in pH values at the end of treatment series, first and second follow-up after treatment with two passes of MDA. CONCLUSIONS: MDA may have remarkable effects on skin barrier function changes resulting in skin clinical improvements (Cochrane Skin Group identifier: CSG No. 37). PMID- 18412874 TI - Combining systemic retinoids with biologic agents for moderate to severe psoriasis. AB - BACKGROUND: Moderate to severe psoriasis, which is defined as psoriasis affecting more than 20% of the body surface area, often requires a combination of therapies to achieve remission. Although numerous data exist regarding the use of acitretin and biologic agent therapy alone for psoriasis, little is known about the efficacy, safety, and tolerability of acitretin combined with biologic agents. METHODS: Fifteen patients with psoriasis treated with concomitant acitretin and a biologic agent were identified, and their charts were reviewed for response to therapy, additional therapy necessary for disease management, side-effects, and laboratory abnormalities whilst on combination therapy. The Institutional Review Board did not require approval for this chart review. RESULTS: Twenty-nine per cent of patients showed clearance of psoriasis, 43% of patients showed an improvement of 90%, 14% showed an improvement of 75%, and 7.1% showed no change. During treatment with acitretin and biologic agent, five patients required no adjunctive treatment. Three patients were able to stop narrow-band ultraviolet-B (UV-B) therapy after an average of 2.33 months of combination therapy. Only one patient continued to require phototherapy (UV-B) in addition to the biologic agent. Three patients developed squamous cell carcinoma (SCC) whilst on combination therapy, but all patients had a previous history of SCC. One patient developed non-Hodgkin's lymphoma after 3 years of etanercept and acitretin, and the etanercept was discontinued. CONCLUSIONS: Acitretin combined with biologic agents offers a promising method of managing refractory psoriasis. More research is needed to determine the long-term safety and efficacy of this combination. PMID- 18412875 TI - Cryosurgery during topical imiquimod: a successful combination modality for lentigo maligna. AB - BACKGROUND: Either cryosurgery or topical imiquimod have been used to treat patients with lentigo maligna in cases where surgery is not feasible. METHODS: We report a patient with lentigo maligna, who was treated with the combination of topical imiquimod and cryosurgery, and review the rationale, which led us to design the present combined cryo-immunological treatment modality. RESULTS: Sustained clearance of lentigo maligna to date (26 months after treatment). The successful treatment of this patient was based on the following rationale: A cryosurgery session during continuing imiquimod application may: (i) reinforce apoptosis of tumor cells; (ii) strengthen antiangiogenesis in the treated tumor; and (iii) build-up a potent tumor-destructive immune response by a cascade of events starting with imiquimod-promoted attraction of immature dendritic antigen presenting cells (DCs) into the tumor. DCs further mature within the tumor antigen-rich environment of subsequently cryo-destructed tumor and upon imiquimod driven migration into the peripheral lymph nodes can stimulate a specific antineoplastic cell-mediated immunity. Finally, continuing imiquimod application after cryosurgery may increase recruitment of activated effector cells into the tumor tissue leading to its destruction. CONCLUSION: Cryosurgery during continued topical imiquimod seems to be a promising treatment for lentigo maligna. PMID- 18412876 TI - Global warming and its dermatologic implications. PMID- 18412877 TI - Chronic cutaneous lupus in childhood: a report of two cases and review of the literature. PMID- 18412878 TI - Treatment of dermal and subcutaneous pheohyphomycosis of 55 years' duration. PMID- 18412879 TI - Fluorescence in situ hybridization analysis is useful for the diagnosis of the carrier state of X-linked ichthyosis. PMID- 18412880 TI - Co-occurrence of nevus comedonicus with accessory breast tissue. PMID- 18412881 TI - Disseminated subcutaneous nodules alone as manifestations of Churg-Strauss syndrome. PMID- 18412883 TI - In the age of professionalism, student harassment is alive and well. PMID- 18412884 TI - Reflection: understanding its influence on practice. PMID- 18412885 TI - Differences in the perceived impact of sleep deprivation among surgical and non surgical residents. AB - OBJECTIVE: Resident work hour restrictions have been mandated in the USA largely out of concern that sleep deprivation compromises doctor performance and patient care. However, individuals' ability to recognise the effects of sleep deprivation has not been studied in medical education. We examined the perceived impact of sleep deprivation among different groups of postgraduate medical trainees. METHODS: A survey addressing work hours, sleepiness and daily functioning was mailed to all residents in the internal medicine, surgery and psychiatry programmes at the University of Toronto who were working at 6 different teaching hospitals. The mailing included the Epworth Sleepiness Scale (ESS), measuring acute sleepiness, and a new Sleep Deprivation Impact (SDI) scale, consisting of 12 items designed to measure the perceived impact of sleep deprivation on an individual's own performance. RESULTS: Overall, 62.5% of surgery (95/152) and 59.5% of non-surgery residents (194/326) completed the survey. Surgery residents reported working longer hours per week (83.0 versus 62.5 hours; P < 0.01), and scored higher on the ESS (12.8 versus 9.2; P < 0.01) compared with other residents. Surgery residents scored significantly lower than others on the SDI scale (45.2 versus 51.5, P < 0.01), indicating less perceived impact of sleep deprivation on performance. CONCLUSIONS: These results are consistent with the presence of an underlying culture within surgery in which individuals may be less willing to accept a natural limitation of individual performance. Whether these findings represent an actual resilience to sleep deprivation among surgery residents or a misperception within this group remains to be determined. PMID- 18412886 TI - Effects of reflective practice on the accuracy of medical diagnoses. AB - CONTEXT: Reflective practice has been suggested to be an important instrument in improving clinical judgement and developing medical expertise. Empirical evidence supporting this suggestion, however, is absent. This paper reports on an experiment conducted to study the effects of reflective practice on diagnostic accuracy. METHODS: Participants were 42 internal medicine residents in hospitals in 2 states in the northeast of Brazil. They diagnosed 16 clinical cases. The experiment employed a repeated measures design, with 2 independent variables: the complexity of clinical cases (simple or complex), and the reasoning approach induced to diagnose the case (participants were instructed to diagnose each case either through pattern recognition or reflective reasoning). The dependent variable was the accuracy of the diagnosis provided for each case. All participants participated in each of the 2 levels of both independent variables. RESULTS: A main effect of case complexity emerged. There was no statistically significant main effect of reflective practice. However, a significant interaction effect was found between case complexity and mode of processing (F[1,41] = 4.48, P < 0.05), indicating that although reflective practice did not make a difference to accuracy of diagnosis in simple cases, it had a positive effect when diagnosing complex cases. CONCLUSIONS: Reflective practice had a positive effect on diagnosis of complex, unusual cases. Non-analytical reasoning was shown to be as effective as reflective reasoning for diagnosing routine clinical cases. Findings support the idea that reflective practice may particularly improve diagnoses in situations of uncertainty and uniqueness, reducing diagnostic errors. PMID- 18412888 TI - Medical education. Introduction. PMID- 18412889 TI - 'Professor, I'm tired and stressed!'. PMID- 18412890 TI - Predicting student performance using artificial neural network analysis. PMID- 18412891 TI - Helping students to gain confidence in personal performance. PMID- 18412892 TI - Preliminary evaluation of a hybrid problem-based learning curriculum. PMID- 18412893 TI - Using interprofessional team-based learning to develop health care policy. PMID- 18412894 TI - Mind the gap! PMID- 18412895 TI - Evaluation of basic science knowledge retention in clinical teaching. PMID- 18412896 TI - Studying published cases and the medical literature curriculum. PMID- 18412897 TI - Changing students' performance in and perception of radiology. PMID- 18412898 TI - 'Who wants to be a neurologist?' An undergraduate quiz. PMID- 18412899 TI - Using OSCEs to teach WHO patient safety solutions. PMID- 18412900 TI - Clinical skills evaluation in a resource-constrained environment. PMID- 18412901 TI - Can cultural competency self-assessment predict OSCE performance? PMID- 18412902 TI - Creating an OSCE/OSPE in a resource-limited setting. PMID- 18412903 TI - Computer-assisted rating of surgical skills: introduction to urate. PMID- 18412905 TI - Online reflective learning supported by student facilitators. PMID- 18412904 TI - A model for the teaching and assessment of professionalism. PMID- 18412906 TI - Learning about plagiarism using Turnitin detection software. PMID- 18412907 TI - The favour: a professional boundaries OSCE station. PMID- 18412908 TI - Stigma: a curriculum to change attitudes, knowledge and skills. PMID- 18412909 TI - Teaching interprofessional health science students medical error disclosure. PMID- 18412910 TI - Using unannounced standardised patients to assess residents' professionalism. PMID- 18412911 TI - Understanding illicit substance use in the real world. PMID- 18412912 TI - Multi-disciplinary service learning: a medico-legal collaboration. PMID- 18412913 TI - Role plays to build counselling competencies. PMID- 18412914 TI - Enhancing communication skills using an OSCE and peer review. PMID- 18412915 TI - Return visits by simulated patients. PMID- 18412916 TI - Improving medical student communication skills through improvisational theatre. PMID- 18412917 TI - Remediation workshop for medical students in patient-doctor interaction skills. PMID- 18412918 TI - Teaching communication skills specific to paediatrics in 40 minutes. PMID- 18412919 TI - Extending the vision: cost-effective relay of real-time simulation. PMID- 18412920 TI - Improving teaching by teaching feedback. PMID- 18412921 TI - Online faculty development for community health in Brazil. PMID- 18412922 TI - Collegial conflict: experiencing attribution theory through simulation. PMID- 18412923 TI - Evaluation of the 'Strongest Link': interactive continuing education. PMID- 18412924 TI - Use of retrospective pre/post assessments in faculty development. PMID- 18412925 TI - ENHANCE: food for the ostriches. PMID- 18412926 TI - Master of nothing but knower of all. PMID- 18412927 TI - Extended-release metformin in Asian patients. PMID- 18412928 TI - Calcium channel blocker-based combination therapy. The best approach in the treatment of high-risk hypertension? PMID- 18412929 TI - A new look at the hormonal and metabolic changes in the ageing male. PMID- 18412930 TI - Lipoproteins: from A to B and maybe C-III. PMID- 18412931 TI - Food, fatness and facts: the need for a standard message. PMID- 18412932 TI - The metabolic effects of once daily extended-release metformin in patients with type 2 diabetes: a multicentre study. AB - AIM: To investigate the effects of extended-release metformin (MXR) compared with immediate-release metformin (MIR) on post-prandial glycaemic excursion, chronic glycaemia, lipid profiles, insulin resistance and islet function in type 2 diabetes. METHODS: A randomised, open-labelled, positive-controlled multicentre study was conducted on 150 Chinese patients with type 2 diabetes. After 2 weeks of run-in period with MIR, 150 subjects were randomised into MXR group and MIR group. The patients in MXR group were assigned to take MXR 1500 mg once daily after dinner, while the patients in MIR group were assigned to continue MIR 500 mg thrice daily after meals for 12 weeks. Standard meal tests were carried out at baseline and at the end of this study. Plasma glucose, serum insulin, HbA1c and lipid profiles were measured. Homeostasis model assessment (HOMA) was used to evaluate insulin resistance index (HOMA-IR) and islet beta-cell function index (HOMA-B). RESULTS: Either MIR or MXR modestly, but significantly decreased HbA1c levels and body mass index (BMI) after 12 weeks of treatment. However, there were no significant differences between two groups. The post-prandial glycaemia at 120 min after a standard meal in MXR group was higher than in MIR group (11.02 +/- 3.08 mmol/l vs. 9.74 +/- 2.61 mmol/l, p < 0.05). Moreover, no differences in the areas under curve of insulin release response, HOMA-B, HOMA-IR and lipid profiles were found within or between groups after 12 weeks of treatment. CONCLUSION: The effects of once daily MXR on chronic glycaemia, BMI, lipid profiles, insulin resistance and islet function are comparable with that of thrice daily MIR in oriental population. PMID- 18412933 TI - A 'one-stop clinic' for the diagnosis and management of rotator cuff pathology: Getting the right diagnosis first time. AB - BACKGROUND: We have introduced a one-stop clinic for the diagnosis and management of rotator cuff tears (RCTs). The aim of the study was to determine the effect of the one-stop clinic on the interval and delay in management for patients with suspected rotator cuff pathology. PATIENTS AND METHODS: Seventy-four patients were reviewed retrospectively; 39 had an ultrasound of the shoulder before (group 1), and 35 patients after (group 2) the new protocol was introduced. Patients in group 2 had a shoulder ultrasound scan (USS) on the day of their clinic appointment or beforehand based upon the General Practitioners (GP) letter. Indications included patients over the age of 30, or patients with a history of trauma or a painful shoulder. RESULTS: Twenty-four patients were diagnosed with RCTs on USS with an overall sensitivity of 93.75%, specificity 100% and accuracy of 91% for full thickness tears. Clinical detection alone had an overall sensitivity of 80%, a specificity of 91% and accuracy of 87%. Mean time from GP referral to definitive management plan was 6.49 months (SD 2.74) in group 1, compared with 4.63 months (SD 1.43) in group 2 with an overall reduction in half the number of clinic appointments. This was statistically significant (p < 0.001). CONCLUSIONS: Ultrasound scan of the shoulder is an accurate and reliable method of detecting full thickness RCTs. The one-stop clinic significantly shortened the interval between GP referral and definitive management. PMID- 18412934 TI - Radiofrequency ablation in the treatment of primary breast cancer: no surgical redundancies yet. AB - BACKGROUND/INTRODUCTION: Radiofrequency energy has emerged as a new tool for the local destruction of cancer by inducing thermal tissue necrosis in the target region. Radiofrequency ablation (RFA) has recently been used to treat breast cancer primaries, potentially offering all the advantages of minimally invasive techniques. METHODS/EVIDENCE: Nine published studies addressing the role of RFA in the treatment of breast cancer have been identified and analysed, in six, first-line RFA was followed by surgical removal and there were 12 failures in 108 ablations. Three further studies involved RFA without subsequent excision and in 1/60 there was a local relapse after 4 months (follow-up range: 15-29 months). DISCUSSION: Existing RFA techniques may not be able to destroy the whole of the malignant lesion, because of local conditions allowing cancer cells to survive within the target area or because electrodes cannot be accurately directed to the tumour site with ultrasound. Additionally, distant in-breast cancers can be missed on pre/intraoperative imaging. Histological information is unavailable after tissue destruction so that the opportunity to reassess tumour grade based on more extensive sampling is lost, but this can be improved by more extensive sampling with vacuum-assisted core biopsy. CONCLUSIONS: Before RFA can be safely used in the treatment of breast cancer primaries, several criteria need to be met. These include development of RFA devices and techniques, standardisation of the treatment protocol, including imaging and selection of patients, and establishment of a feasible post-treatment follow-up strategy. PMID- 18412935 TI - Testicular torsion: a race against time. AB - Torsion of the testis is a medical emergency that is most commonly encountered in adolescents. Patients usually present with sudden onset scrotal pain associated with nausea and vomiting. On physical examination the involved testis is tender, high riding and usually horizontal. The cremasteric reflex is usually absent. If left untreated irreversible ischaemia starts appearing in 6 h. Doppler ultrasound is the diagnostic imaging of choice. The ideal treatment is surgical exploration and orchidectomy with contralateral orchidopexy or bilateral orchidopexy depending on the condition of the affected testis. If surgical options are delayed then manual detorsion should be attempted. PMID- 18412936 TI - Is there any link between cardiovascular disease and depression? PMID- 18412937 TI - The first report of Prosopis farcta ingestion in children: is it serious? PMID- 18412938 TI - Trigeminal neuralgia as a presentation of infective endocarditis. PMID- 18412939 TI - The Joint British Societies' algorithm for the diagnosis of diabetes mellitus in routine clinical practice. AB - The diagnosis of diabetes mellitus (DM) requires a logical approach to identify new cases while not subjecting every patient to an oral glucose tolerance test (OGTT). The Joint British Societies' (JBS2) have recommended an algorithm for the diagnosis of DM. We retrospectively obtained the results of 100 OGTTs requested by primary care physicians, along with previous glucose results and whether the patient was fasting or not with a view to determining how many OGTTs could be avoided by following the recommended algorithm. We found that 47% of the OGTTs could have been avoided, of which the algorithm would have identified all but 4%. Two cases of DM and eight cases of impaired glucose tolerance (IGT) would have been missed. We conclude that following the JBS2 algorithm for the diagnosis of DM will significantly reduce the number of OGTTs, with only a few patients with IGT or DM being missed when such a policy is used. PMID- 18412940 TI - Interpreting and applying the EUFEST results using number needed to treat: antipsychotic effectiveness in first-episode schizophrenia. PMID- 18412942 TI - Analysis of bacterial DNA in synovial tissue of Tunisian patients with reactive and undifferentiated arthritis by broad-range PCR, cloning and sequencing. AB - INTRODUCTION: Bacteria and/or their antigens have been implicated in the pathogenesis of reactive arthritis (ReA). Several studies have reported the presence of bacterial antigens and nucleic acids of bacteria other than those specified by diagnostic criteria for ReA in joint specimens from patients with ReA and various arthritides. The present study was conducted to detect any bacterial DNA and identify bacterial species that are present in the synovial tissue of Tunisian patients with reactive arthritis and undifferentiated arthritis (UA) using PCR, cloning and sequencing. METHODS: We examined synovial tissue samples from 28 patients: six patients with ReA and nine with UA, and a control group consisting of seven patients with rheumatoid arthritis and six with osteoarthritis (OA). Using broad-range bacterial PCR producing a 1,400-base-pair fragment from the 16S rRNA gene, at least 24 clones were sequenced for each synovial tissue sample. To identify the corresponding bacteria, DNA sequences were compared with sequences from the EMBL (European Molecular Biology Laboratory) database. RESULTS: Bacterial DNA was detected in 75% of the 28 synovial tissue samples. DNA from 68 various bacterial species were found in ReA and UA samples, whereas DNA from 12 bacteria were detected in control group samples. Most of the bacterial DNAs detected were from skin or intestinal bacteria. DNA from bacteria known to trigger ReA, such as Shigella flexneri and Shigella sonnei, were detected in ReA and UA samples of synovial tissue and not in control samples. DNA from various bacterial species detected in this study have not previously been found in synovial samples. CONCLUSION: This study is the first to use broad-range PCR targeting the full 16S rRNA gene for detection of bacterial DNA in synovial tissue. We detected DNA from a wide spectrum of bacterial species, including those known to be involved in ReA and others not previously associated with ReA or related arthritis. The pathogenic significance of some of these intrasynovial bacterial DNAs remains unclear. PMID- 18412943 TI - Investment in online self-evaluation tests: A theoretical approach. AB - BACKGROUND: Large-scale traumatic events may burden any affected public health system with consequential charges. One major post-disaster, expense factor emerges form early psychological interventions and subsequent, posttraumatic mental health care. Due to the constant increase in mental health care costs, also post-disaster public mental health requires best possible, cost-effective care systems. Screening and monitoring the affected population might be one such area to optimize the charges. METHODS: This paper analyzes the potential cost effectiveness of monitoring a psychologically traumatized population and to motivate individuals at risk to seek early treatment. As basis for our model served Grossman's health production function, which was modified according to fundamental concepts of cost-benefit analyzes, to match the basic conditions of online monitoring strategies. We then introduce some fundamental concepts of cost benefit analysis. RESULTS: When performing cost-benefit analyses, policy makers have to consider both direct costs (caused by treatment) and indirect costs (due to non-productivity). Considering both costs sources we find that the use of Internet-based psychometric screening instruments may reduce the duration of future treatment, psychological burden and treatment costs. CONCLUSION: The identification of individuals at risk for PTSD following a disaster may help organizations prevent both the human and the economic costs of this disease. Consequently future research on mental health issues should put more emphasis on the importance of monitoring to detect early PTSD and focus the most effective resources within early treatment and morbidity prevention. PMID- 18412944 TI - Design of a placebo-controlled, randomized study of the efficacy of repetitive transcranial magnetic stimulation for the treatment of chronic tinnitus. AB - BACKGROUND: Chronic tinnitus is a frequent condition, which can have enormous impact on patient's life and which is very difficult to treat. Accumulating data indicate that chronic tinnitus is related to dysfunctional neuronal activity in the central nervous system. Repetitive transcranial magnetic stimulation (rTMS) is a non-invasive method which allows to focally modulate neuronal activity. An increasing amount of studies demonstrate reduction of tinnitus after repeated sessions of low-frequency rTMS and indicate that rTMS might represent a new promising approach for the treatment of tinnitus. However available studies have been mono-centric and are characterized by small sample sizes. Therefore, this multi-center trial will test the efficacy of rTMS treatment in a large sample of chronic tinnitus patients. METHODS/DESIGN: This is a randomized, placebo controlled, double-blind multi-center trial of two weeks 1 Hz rTMS-treatment in chronic tinnitus patients. Eligible patients will be randomized to either 2 weeks real or sham rTMS treatment. Main eligibility criteria: male or female individuals aged 18-70 years with chronic tinnitus (duration > 6 months), tinnitus-handicap-inventory-score > or = 38, age-adjusted normal sensorineural hearing (i.e. not more than 5 dB below the 10% percentile of the appropriate age and gender group (DIN EN ISO 7029), conductive hearing loss < or = 15dB. The primary endpoint is a change of tinnitus severity according to the tinnitus questionnaire of Goebel and Hiller (baseline vs. end of treatment period). A total of 138 patients are needed to detect a clinical relevant change of tinnitus severity (i.e. 5 points on the questionnaire of Goebel and Hiller; alpha = 0.05; 1-beta = 0.80). Assuming a drop-out rate of less than 5% until the primary endpoint, 150 patients have to be randomized to guarantee the target number of 138 evaluable patients. The study will be conducted by otorhinolaryngologists and psychiatrists of 7 university hospitals and 1 municipal hospital in Germany. DISCUSSION: This study will provide important information about the efficacy of rTMS in the treatment of chronic tinnitus. TRIAL REGISTRATION: Current Controlled Trials ISRCTN89848288. PMID- 18412945 TI - Pharmacokinetic characteristics and anticancer effects of 5-fluorouracil loaded nanoparticles. AB - BACKGROUND: It is expected that prolonged circulation of anticancer drugs will increase their anticancer activity while decreasing their toxic side effects. The purpose of this study was to prepare 5-fluorouracil (5-FU) loaded block copolymers, with poly(gamma-benzyl-L-glutamate) (PBLG) as the hydrophobic block and poly(ethylene glycol) (PEG) as the hydrophilic block, and then examine the 5 FU release characteristics, pharmacokinetics, and anticancer effects of this novel compound. METHODS: 5-FU loaded PEG-PBLG (5-FU/PEG-PBLG) nanoparticles were prepared by dialysis and then scanning electron microscopy (SEM) and transmission electron microscopy (TEM) were used to observe the shape and size of the nanoparticles, and ultraviolet spectrophotometry was used to evaluate the 5-FU in vitro release characteristics. The pharmacokinetic parameters of 5-FU/PEG-PBLG nanoparticles in rabbit plasma were determined by measuring the 5-FUby high performance liquid chromatography (HPLC). To study in vivo effects, LoVo cells (human colon cancer cell line) or Tca8113 cells (human oral squamous cell carcinoma cell line) were implanted in BALB/c nude mice that were subsequently treated with 5-FU or 5-FU/PEG-PBLG nanospheres. RESULTS: 5-FU/PEG-PBLG nanoparticles had a core-shell spherical structure with a diameter of 200 nm and a shell thickness of 30 nm. The drug loading capacity was 27.1% and the drug encapsulation was 61.5%. Compared with 5-FU, 5-FU/PEG-PBLG nanoparticles had a longer elimination half-life (t1/2, 33.3 h vs. 5 min), lower peak concentration (C, 4563.5 microg/L vs. 17047.3 microg/L), and greater distribution volume (VD, 0.114 L vs. 0.069 L). Compared with a blank control, LoVo cell xenografts and Tca8113 cell xenografts treated with 5-FU or 5-FU/PEG-PBLG nanoparticles grew slower and had prolonged tumor doubling times. 5-FU/PEG-PBLG nanoparticles showed greater inhibition of tumor growth than 5-FU (p < 0.01). In the PEG-PBLG nanoparticle control group, there was no tumor inhibition (p > 0.05). CONCLUSION: In our model system, 5-FU/PEG-PBLG nanoparticles changed the pharmacokinetic behavior of 5-FU, thus increasing its anticancer activity. 5-Fluorouracil loaded nanoparticles have potential as a novel anticancer drug that may have useful clinical applications. PMID- 18412946 TI - Quantifying the effectiveness of climate change mitigation through forest plantations and carbon sequestration with an integrated land-use model. AB - BACKGROUND: Carbon plantations are introduced in climate change policy as an option to slow the build-up of atmospheric carbon dioxide (CO2) concentrations. Here we present a methodology to evaluate the potential effectiveness of carbon plantations. The methodology explicitly considers future long-term land-use change around the world and all relevant carbon (C) fluxes, including all natural fluxes. Both issues have generally been ignored in earlier studies. RESULTS: Two different baseline scenarios up to 2100 indicate that uncertainties in future land-use change lead to a near 100% difference in estimates of carbon sequestration potentials. Moreover, social, economic and institutional barriers preventing carbon plantations in natural vegetation areas decrease the physical potential by 75-80% or more.Nevertheless, carbon plantations can still considerably contribute to slowing the increase in the atmospheric CO2 concentration but only in the long term. The most conservative set of assumptions lowers the increase of the atmospheric CO2 concentration in 2100 by a 27 ppm and compensates for 5-7% of the total energy-related CO2 emissions. The net sequestration up to 2020 is limited, given the short-term increased need for agricultural land in most regions and the long period needed to compensate for emissions through the establishment of the plantations. The potential is highest in the tropics, despite projections that most of the agricultural expansion will be in these regions. Plantations in high latitudes as Northern Europe and Northern Russia should only be established if the objective to sequester carbon is combined with other activities. CONCLUSION: Carbon sequestration in plantations can play an important role in mitigating the build-up of atmospheric CO2. The actual magnitude depends on natural and management factors, social barriers, and the time frame considered. In addition, there are a number of ancillary benefits for local communities and the environment. Carbon plantations are, however, particularly effective in the long term. Furthermore, plantations do not offer the ultimate solution towards stabilizing CO2 concentrations but should be part of a broader package of options with clear energy emission reduction measures. PMID- 18412947 TI - Differential prefrontal-like deficit in children after cerebellar astrocytoma and medulloblastoma tumor. AB - BACKGROUND: This study was realized thanks to the collaboration of children and adolescents who had been resected from cerebellar tumors. The medulloblastoma group (CE+, n = 7) in addition to surgery received radiation and chemotherapy. The astrocytoma group (CE, n = 13) did not receive additional treatments. Each clinical group was compared in their executive functioning with a paired control group (n = 12). The performances of the clinical groups with respect to controls were compared considering the tumor's localization (vermis or hemisphere) and the affectation (or not) of the dentate nucleus. Executive variables were correlated with the age at surgery, the time between surgery-evaluation and the resected volume. METHODS: The executive functioning was assessed by means of WCST, Complex Rey Figure, Controlled Oral Word Association Test (letter and animal categories), Digits span (WISC-R verbal scale) and Stroop test. These tests are very sensitive to dorsolateral PFC and/or to medial frontal cortex functions. The scores for the non-verbal Raven IQ were also obtained. Direct scores were corrected by age and transformed in standard scores using normative data. The neuropsychological evaluation was made at 3.25 (SD = 2.74) years from surgery in CE group and at 6.47 (SD = 2.77) in CE+ group. RESULTS: The Medulloblastoma group showed severe executive deficit (0.30). For ICU length of stay (LoS) of greater than or equal to 3 days, hospital mortality was reduced from 34.1% to 25.4% (-26%) (P = 0.05). For ICU LoS of greater than or equal to 4 days, hospital mortality was reduced from 34.3% to 23.5% (-32%) (P = 0.02). For ICU LoS of greater than or equal to 5 days, hospital mortality was reduced from 31.9% to 20.6% (-35%) (P = 0.02). ICU mortality was also reduced but the P value was less than 0.13 for ICU LoS of greater than or equal to 4 and 5 days. CONCLUSION: SPRINT achieved a high level of glycaemic control on a severely ill critical cohort population. Reductions in mortality were observed compared with a retrospective hyperglycaemic cohort. Range and peak blood glucose metrics were no longer correlated with mortality outcome under SPRINT. PMID- 18412979 TI - Risk factors for antenatal depression, postnatal depression and parenting stress. AB - BACKGROUND: Given that the prevalence of antenatal and postnatal depression is high, with estimates around 13%, and the consequences serious, efforts have been made to identify risk factors to assist in prevention, identification and treatment. Most risk factors associated with postnatal depression have been well researched, whereas predictors of antenatal depression have been less researched. Risk factors associated with early parenting stress have not been widely researched, despite the strong link with depression. The aim of this study was to further elucidate which of some previously identified risk factors are most predictive of three outcome measures: antenatal depression, postnatal depression and parenting stress and to examine the relationship between them. METHODS: Primipara and multiparae women were recruited antenatally from two major hoitals as part of the beyondblue National Postnatal Depression Program 1. In this subsidiary study, 367 women completed an additional large battery of validated questionnaires to identify risk factors in the antenatal period at 26-32 weeks gestation. A subsample of these women (N = 161) also completed questionnaires at 10-12 weeks postnatally. Depression level was measured by the Beck Depression Inventory (BDI). RESULTS: Regression analyses identified significant risk factors for the three outcome measures. (1). Significant predictors for antenatal depression: low self-esteem, antenatal anxiety, low social support, negative cognitive style, major life events, low income and history of abuse. (2). Significant predictors for postnatal depression: antenatal depression and a history of depression while also controlling for concurrent parenting stress, which was a significant variable. Antenatal depression was identified as a mediator between seven of the risk factors and postnatal depression. (3). Postnatal depression was the only significant predictor for parenting stress and also acted as a mediator for other risk factors. CONCLUSION: Risk factor profiles for antenatal depression, postnatal depression and parenting stress differ but are interrelated. Antenatal depression was the strongest predictor of postnatal depression, and in turn postnatal depression was the strongest predictor for parenting stress. These results provide clinical direction suggesting that early identification and treatment of perinatal depression is important. PMID- 18412980 TI - Effects of pectin liquid on gastroesophageal reflux disease in children with cerebral palsy. AB - BACKGROUND: The use of thickeners is a standard therapy for decreasing episodes of regurgitation or vomiting in infants. However, it remains to be investigated whether thickener is effective for vomiting and/or chronic respiratory symptoms in children with cerebral palsy. METHODS: We enrolled 18 neurologically impaired children caused by cerebral palsy, with gastroesophageal reflux disease. In the first part of this study (pH monitoring), subjects were randomly allocated to two groups: fed with a high-pectin diet [enteral formula: pectin liquid = 2:1 (v/v)], or a low-pectin diet [enteral formula: pectin liquid = 3:1 (v/v)]. Two-channel esophageal pH monitoring was performed over 48 h. In the second part (clinical trial), subjects were fed a high- or low-pectin diet and non-pectin diet for 4 weeks in a crossover manner. Nurses recorded the feeding volume, number of episodes of vomiting, volume of gastric residue, episodes of cough and wheeze, frequency of using oxygen for dyspnea, and the day when the child could return to school. Cough and wheeze were recorded as a cough-score. RESULTS: The median value for the % time pH < 4 at the lower and upper esophagus was significantly decreased with a high-pectin diet [9.2% (6.2-22.6) vs. 5.0% (3.1-13.1); P < 0.01, 3.8% (2.9-11.2) vs. 1.6% (0.9-8.9); P < 0.01 (interquartile range), non-pectin and high-pectin, respectively]. The number of reflux episodes per day and duration of longest reflux were decreased significantly with a high-pectin, but not with a low-pectin diet. The median number of episodes of vomiting decreased significantly with a high-pectin diet [2.5/week (1.0-5.0) vs. 1.0 (1.0-1.5), P < 0.05]. The median cough-score was significantly decreased by both concentrations of pectin [8.5/week (1.0-11.5) vs. 2.0/week (0.0-3.0), fed with a high-pectin diet; 7.0/week (1.0-14.5) vs. 1.0/w (0.0-5.0), fed with a low-pectin diet, P < 0.05]. CONCLUSION: Pectin liquid partially decreased gastroesophageal reflux as measured by eshophageal pH monitoring, and might improve vomiting and respiratory symptoms in children with cerebral palsy. TRIAL REGISTRATION: ISRCTN19787793. PMID- 18412981 TI - Plasma intact fibroblast growth factor 23 levels in women with anorexia nervosa. AB - BACKGROUND: Fibroblast growth factor (FGF)23 is a novel phosphaturic factor associated with inorganic phosphate homeostasis. Previous human studies have shown that serum FGF23 levels increase in response to a high phosphate diet. For anorexia nervosa (AN) patients, inorganic phosphate homeostasis is important in the clinical course, such as in refeeding syndrome. The purpose of this study was to determine plasma levels of intact FGF23 (iFGF23) in restricting-type AN (AN-R) patients, binge-eating/purging-type AN (AN-BP) patients, and healthy controls. METHODS: The subjects consisted of 6 female AN-R patients, 6 female AN-BP patients, and 11 healthy female controls; both inpatients and outpatients were included. Plasma iFGF23, 1,25-dihydroxyvitamin D (1,25-(OH)2D), and 25 hydroxyvitamin D (25-OHD) levels were measured. Data are presented as the median and the range. A two-tailed Mann-Whitney U-test with Bonferroni correction was used to assess differences among the three groups, and a value of p < 0.017 was considered statistically significant. RESULTS: There were no differences between AN-R patients and controls in the iFGF23 and 1,25-(OH)2D levels. In AN-BP patients, the iFGF23 level (41.3 pg/ml; range, 6.1-155.5 pg/ml) was significantly higher than in controls (3.8 pg/ml; range, not detected-21.3 pg/ml; p = 0.001), and the 1,25-(OH)2D was significantly lower in AN-BP patients (7.0 pg/ml; range, 4.2-33.7 pg/ml) than in controls (39.7 pg/ml; range, 6.3-58.5 pg/ml; p = 0.015). No differences in plasma 25-OHD levels were observed among the groups. CONCLUSION: This preliminary study is the first to show that plasma iFGF23 levels are increased in AN-BP patients, and that these elevated plasma FGF23 levels might be related to the decrease in plasma 1,25-(OH)2D levels. PMID- 18412982 TI - Supernormal functional reserve of apical segments in elite soccer players: an ultrasound speckle tracking handgrip stress study. AB - BACKGROUND: Ultrasound speckle tracking from grey scale images allows the assessment of regional strain derived from 2D regardless of angle intonation, and it is highly reproducible. The study aimed to evaluate regional left ventricular functional reserve in elite soccer players. METHODS: 50 subjects (25 elite athletes and 25 sedentary controls), aged 26 +/- 3.5, were submitted to an echo exam, at rest and after the Hand Grip (HG) test. Both standard echo parameters and strain were evaluated. RESULTS: Ejection fraction was similar in athletes and controls both at rest (athletes 58 +/- 2 vs controls 57 +/- 4 p ns) and after HG (athletes 60 +/- 2 vs controls 58 +/- 3 p ns). Basal (septal and anterior) segments showed similar strain values in athletes and controls both at rest (athletes S% -19.9 +/- 4.2; controls S% -18.8 +/- 4.9 p = ns) and after HG (athletes S% -20.99 +/- 2.8; controls S% -19.46 +/- 4.4 p = ns). Medium-apical segments showed similar strain values at rest (athletes S% -17.31 +/- 2.3; controls S% -20.00 +/- 5.3 p = ns), but higher values in athletes after HG (athletes S% -24.47 +/- 2.8; controls S% -20.47 +/- 5.4 p < 0.05) CONCLUSION: In athletes with physiological myocardial hypertrophy, a brief isometric effort produces enhancement of the strain in medium-apical left ventricular segments, suggesting the presence of a higher regional function reserve which can be elicited with an inotropic challenge and suitable methods of radial function quantification such as 2D-derived strain. PMID- 18412983 TI - Multiway real-time PCR gene expression profiling in yeast Saccharomyces cerevisiae reveals altered transcriptional response of ADH-genes to glucose stimuli. AB - BACKGROUND: The large sensitivity, high reproducibility and essentially unlimited dynamic range of real-time PCR to measure gene expression in complex samples provides the opportunity for powerful multivariate and multiway studies of biological phenomena. In multiway studies samples are characterized by their expression profiles to monitor changes over time, effect of treatment, drug dosage etc. Here we perform a multiway study of the temporal response of four yeast Saccharomyces cerevisiae strains with different glucose uptake rates upon altered metabolic conditions. RESULTS: We measured the expression of 18 genes as function of time after addition of glucose to four strains of yeast grown in ethanol. The data are analyzed by matrix-augmented PCA, which is a generalization of PCA for 3-way data, and the results are confirmed by hierarchical clustering and clustering by Kohonen self-organizing map. Our approach identifies gene groups that respond similarly to the change of nutrient, and genes that behave differently in mutant strains. Of particular interest is our finding that ADH4 and ADH6 show a behavior typical of glucose-induced genes, while ADH3 and ADH5 are repressed after glucose addition. CONCLUSION: Multiway real-time PCR gene expression profiling is a powerful technique which can be utilized to characterize functions of new genes by, for example, comparing their temporal response after perturbation in different genetic variants of the studied subject. The technique also identifies genes that show perturbed expression in specific strains. PMID- 18412984 TI - Transcriptional regulation by poly(ADP-ribose) polymerase-1 during T cell activation. AB - BACKGROUND: Accumulating evidence suggests an important role for the enzyme poly(ADP-ribose) polymerase-1 (PARP-1) as an integral part of the gene expression regulatory machinery during development and in response to specific cellular signals. PARP-1 might modulate gene expression through its catalytic activity leading to poly(ADP-ribosyl)ation of nuclear proteins or by its physical association with relevant proteins. Recently, we have shown that PARP-1 is activated during T cell activation. However, the proposed role of PARP-1 in reprogramming T cell gene expression upon activation remains largely unexplored. RESULTS: In the present study we use oligonucleotide microarray analysis to gain more insight into the role played by PARP-1 during the gene expression reprogramming that takes place in T cells upon activation with anti-CD3 stimulation alone, or in combination with anti-CD28 co-stimulation. We have identified several groups of genes with expression modulated by PARP-1. The expression of 129 early-response genes to anti-CD3 seems to be regulated by PARP 1 either in a positive (45 genes) or in a negative manner (84 genes). Likewise, in the presence of co-stimulation (anti-CD3 + anti-CD28 stimulation), the expression of 203 genes is also regulated by PARP-1 either up (173 genes) or down (30 genes). Interestingly, PARP-1 deficiency significantly alters expression of genes associated with the immune response such as chemokines and genes involved in the Th1/Th2 balance. CONCLUSION: This study provides new insights into changes in gene expression mediated by PARP-1 upon T cell activation. Pathway analysis of PARP-1 as a nuclear signalling molecule in T cells would be of relevance for the future development of new therapeutic approaches targeting PARP-1 in the acquired immune response. PMID- 18412985 TI - Imp-L2, a putative homolog of vertebrate IGF-binding protein 7, counteracts insulin signaling in Drosophila and is essential for starvation resistance. AB - BACKGROUND: Insulin and insulin-like growth factors (IGFs) signal through a highly conserved pathway and control growth and metabolism in both vertebrates and invertebrates. In mammals, insulin-like growth factor binding proteins (IGFBPs) bind IGFs with high affinity and modulate their mitogenic, anti apoptotic and metabolic actions, but no functional homologs have been identified in invertebrates so far. RESULTS: Here, we show that the secreted Imaginal morphogenesis protein-Late 2 (Imp-L2) binds Drosophila insulin-like peptide 2 (Dilp2) and inhibits growth non-autonomously. Whereas over-expressing Imp-L2 strongly reduces size, loss of Imp-L2 function results in an increased body size. Imp-L2 is both necessary and sufficient to compensate Dilp2-induced hyperinsulinemia in vivo. Under starvation conditions, Imp-L2 is essential for proper dampening of insulin signaling and larval survival. CONCLUSION: Imp-L2, the first functionally characterized insulin-binding protein in invertebrates, serves as a nutritionally controlled suppressor of insulin-mediated growth in Drosophila. Given that Imp-L2 and the human tumor suppressor IGFBP-7 show sequence homology in their carboxy-terminal immunoglobulin-like domains, we suggest that their common precursor was an ancestral insulin-binding protein. PMID- 18412986 TI - The vitamin D receptor in cancer. AB - Over the last 25 years roles have been established for vitamin D receptor (VDR) in influencing cell proliferation and differentiation. For example, murine knock out approaches have revealed a role for the VDR in controlling mammary gland growth and function. These actions appear widespread, as the enzymes responsible for 1alpha,25-dihydroxycholecalciferol generation and degradation, and the VDR itself, are all functionally present in a wide range of epithelial and haematopoietic cell types. These findings, combined with epidemiological and functional data, support the concept that local, autocrine and paracrine VDR signalling exerts control over cell-fate decisions in multiple cell types. Furthermore, the recent identification of bile acid lithocholic acid as a VDR ligand underscores the environmental sensing role for the VDR. In vitro and in vivo dissection of VDR signalling in cancers (e.g. breast, prostate and colon) supports a role for targeting the VDR in either chemoprevention or chemotherapy settings. As with other potential therapeutics, it has become clear that cancer cells display de novo and acquired genetic and epigenetic mechanisms of resistance to these actions. Consequently, a range of experimental and clinical options are being developed to bring about more targeted actions, overcome resistance and enhance the efficacy of VDR-centred therapeutics. PMID- 18412987 TI - Obesity and cancer. AB - The prevalence of obesity, defined as a BMI of > or =30.0 kg/m2, has increased substantially over previous decades to about 20% in industrialized countries, and a further increase is expected in the future. Epidemiological studies have shown that obesity is a risk factor for: post-menopausal breast cancer; cancers of the endometrium, colon and kidney; malignant adenomas of the oesophagus. Obese subjects have an approximately 1.5-3.5-fold increased risk of developing these cancers compared with normal-weight subjects, and it has been estimated that between 15 and 45% of these cancers can be attributed to overweight (BMI 25.0 29.9 kg/m2) and obesity in Europe. More recent studies suggest that obesity may also increase the risk of other types of cancer, including pancreatic, hepatic and gallbladder cancer. The underlying mechanisms for the increased cancer risk as a result of obesity are unclear and may vary by cancer site and also depend on the distribution of body fat. Thus, abdominal obesity as defined by waist circumference or waist:hip ratio has been shown to be more strongly related to certain cancer types than obesity as defined by BMI. Possible mechanisms that relate obesity to cancer risk include insulin resistance and resultant chronic hyperinsulinaemia, increased production of insulin-like growth factors or increased bioavailability of steroid hormones. Recent research also suggests that adipose tissue-derived hormones and cytokines (adipokines), such as leptin, adiponectin and inflammatory markers, may reflect mechanisms linked to tumourigenesis. PMID- 18412988 TI - Genome-health nutrigenomics and nutrigenetics: nutritional requirements or 'nutriomes' for chromosomal stability and telomere maintenance at the individual level. AB - It is becoming increasingly evident that (a) risk for developmental and degenerative disease increases with more DNA damage, which in turn is dependent on nutritional status, and (b) the optimal concentration of micronutrients for prevention of genome damage is also dependent on genetic polymorphisms that alter the function of genes involved directly or indirectly in the uptake and metabolism of micronutrients required for DNA repair and DNA replication. The development of dietary patterns, functional foods and supplements that are designed to improve genome-health maintenance in individuals with specific genetic backgrounds may provide an important contribution to an optimum health strategy based on the diagnosis and individualised nutritional prevention of genome damage, i.e. genome health clinics. The present review summarises some of the recent knowledge relating to micronutrients that are associated with chromosomal stability and provides some initial insights into the likely nutritional factors that may be expected to have an impact on the maintenance of telomeres. It is evident that developing effective strategies for defining nutrient doses and combinations or 'nutriomes' for genome-health maintenance at the individual level is essential for further progress in this research field. PMID- 18412989 TI - Biomarkers of bone health and osteoporosis risk. AB - The assay features of biochemical markers of bone turnover have markedly improved in the past few years. The most sensitive and specific markers of bone formation include serum bone alkaline phosphatase, total osteocalcin (including the intact molecule and the large N-mid fragment) and the procollagen type I N-terminal propeptide assay. Among the various markers of bone resorption, measurements of the urinary excretion of N- and C-terminal cross-linked telopeptides) and of serum C-terminal cross-linked telopeptides are the most sensitive and specific. Markers of bone turnover can be used to predict the rate of bone loss in post menopausal women and can also be used to assess the risk of fractures. In osteoporosis-treatment studies (with alendronate, risedronate, raloxifene) markers of bone turnover appear even more strongly associated with fracture risk reduction than bone mineral density (BMD). These observations support the use of markers of bone turnover as surrogates for fracture risk reduction, perhaps even more so than BMD. Bone markers can also be used to monitor the efficacy of antiresorptive therapy such as hormone-replacement therapy, raloxifene and bisphosphonates in individual patients. Furthermore, they have also proved to be helpful in monitoring the response to nutritional interventions and have the advantage over BMD in that they provide information about mechanism of effect and changes are often observed much more rapidly. PMID- 18412990 TI - Importance of calcium, vitamin D and vitamin K for osteoporosis prevention and treatment. AB - Throughout the life cycle the skeleton requires optimum development and maintenance of its integrity to prevent fracture. Bones break because the loads placed on them exceed the ability of the bone to absorb the energy involved. It is now estimated that one in three women and one in twelve men aged >55 years will suffer from osteoporosis in their lifetime and at a cost in the UK of > 1.7 pounds x 10(9) per year. The pathogenesis of osteoporosis is multifactorial. Both the development of peak bone mass and the rate of bone loss are determined by key endogenous and exogenous factors. Ca supplements appear to be effective in reducing bone loss in women late post menopause (>5 years post menopause), particularly in those with low habitual Ca intake (<400 mg/d). In women early post menopause (<5 years post menopause) who are not vitamin D deficient, Ca supplementation has little effect on bone mineral density. However, supplementation with vitamin D and Ca has been shown to reduce fracture rates in the institutionalised elderly, but there remains controversy as to whether supplementation is effective in reducing fracture in free-living populations. Re defining vitamin D requirements in the UK is needed since there is evidence of extensive hypovitaminosis D in the UK. Low vitamin D status is associated with an increased risk of falling and a variety of other health outcomes and is an area that requires urgent attention. The role of other micronutrients on bone remains to be fully defined, although there are promising data in the literature for a clear link between vitamin K nutrition and skeletal integrity, including fracture reduction. PMID- 18412991 TI - Nutritional factors affecting poultry bone health. AB - Outlined are two main current research concerns relating to skeletal disorders in poultry: (a) osteoporosis in egg-laying hens; (b) leg problems caused by rapid bone growth in broiler chickens. Surveys indicate that 30% of caged laying hens suffer at least one lifetime fracture (a severe welfare issue). Modern hybrids produce one egg per d for 50 weeks. For this period 'normal' bone turnover ceases; only medullary bone (MB) is formed, a woven bone type of limited structural value. MB is resorbed for eggshell formation alongside structural bone, leading to increased fracture risk. Avian osteoporosis is reduced by activity and genetic selection but nutrition is also important. Fluoride and vitamin K are beneficial but the timing of nutritional intervention is important. Ca, inorganic P and vitamin D must be adequate and the form of Ca is critical. Limestone fed as particulates benefits skeletal and eggshell quality. In hens fed particulate limestone compared with flour-fed hens the tibiotarsus breaking strength and radiographic density are increased at 56 weeks of age (P<0.01 and P<0.001 respectively) and the number of tartrate-resistant acid phosphatase positive stained active osteoclasts (mean number per microscopic field) is decreased (P<0.001). In broiler (meat) chickens selection for rapid growth from approximately 50 g to 3 kg in 42 d has inadvertently produced skeletal disorders such as tibial dyschondroplasia, rickets and associated valgus-varus deformities leading to lameness. The beneficial skeletal effects during growth of increased dietary n-3 PUFA:n-6 PUFA (utilising salmon oil) have been demonstrated. Experiments simulating daylight UVB levels have produced beneficial skeletal effects in Ca- and vitamin D-deficient chicks. PMID- 18412992 TI - Phyto-oestrogens and bone health. AB - As oestrogen deficiency is the main cause in the pathogenesis of osteoporosis hormone-replacement therapy remains the mainstay for prevention. However, prophylaxis by hormone-replacement therapy is limited. Phyto-oestrogens, which are weakly-oestrogenic compounds present in plants, deserve particular mention because emerging data support the suggestion that they may prevent bone loss associated with the menopause. In the past few years extensive research using animal models has provided convincing data to indicate a significant improvement in bone mass or other end points following feeding with soyabean. Moreover, observational studies relate the lower incidence of osteoporosis among women in the Eastern world to a diet rich in phyto-oestrogens. However, it is not valid to extrapolate to the Western situation. The varied clinical trials that have been published suggest that isoflavones reduce bone loss in women in the early period post menopause, but a definitive result requires more investigations of the effect of phyto-oestrogens on bone health that have substantial sample size and are of long duration. In addition, the clinical efficacy of soya foods in preventing osteopenia depends on their intestinal metabolism. Thus, phyto oestrogens are a source for putative innovative dietary health intervention for post-menopausal women. However, more data are necessary, particularly in relation to their effect on the risk of fracture. PMID- 18412993 TI - Altered bone metabolism in inflammatory disease: role for nutrition. AB - Osteoporosis is a major public health problem, and as life expectancy and the world's population continue to increase will become even more important. Thus, there is an urgent need to develop and implement nutritional approaches and policies for the prevention and treatment of osteoporosis. Patients with some chronic inflammatory diseases appear to be more likely to develop osteopenia, and in some cases earlier in life, which is of particular concern as the incidence of inflammatory diseases in the Western world is increasing. While the cause of bone loss in patients with inflammatory disease is multifactorial, nutrition may have a role. Many of these patients may have one or more nutritional deficiencies, which can lead to altered rates of bone metabolism. On the other hand, some nutritional factors may attenuate the inflammatory process itself, and thus may indirectly benefit bone metabolism and bone health in patients with inflammatory disease. The present review will consider these issues, particularly in the context of inflammatory bowel disease, coeliac disease and atherosclerosis. PMID- 18412994 TI - Nutrigenetics and CVD: what does the future hold? AB - CVD is a common killer in both the Western world and the developing world. It is a multifactorial disease that is influenced by many environmental and genetic factors. Although public health advice to date has been principally in the form of prescribed population-based recommendations, this approach has been surprisingly unsuccessful in reducing CVD risk. This outcome may be explained, in part, by the extreme variability in response to dietary manipulations between individuals and interactions between diet and an individual's genetic background, which are defined by the term 'nutrigenetics'. The shift towards personalised nutritional advice is a very attractive proposition. In principle an individual could be genotyped and given dietary advice specifically tailored to their genetic make-up. Evidence-based research into interactions between fixed genetic variants, nutrient intake and biomarkers of CVD risk is increasing, but still limited. The present paper will review the evidence for interactions between dietary fat and three common polymorphisms in the apoE, apoAI and PPARgamma genes. Increased knowledge of how these and other genes influence dietary response should increase the understanding of personalised nutrition. While targeted dietary advice may have considerable potential for reducing CVD risk, the ethical issues associated with its routine use need careful consideration. PMID- 18412995 TI - Antioxidants and CVD. AB - The involvement of free radicals and reactive oxygen and nitrogen species in the pathology of inflammatory and degenerative disease has been widely accepted, although the centrality of these species to the outcome of these diseases is still a matter for debate. In the case of CVD, and particularly the development of the atherosclerotic plaque, the oxidation of LDL is of particular importance and appears to explain many of the events that occur during the life history of the plaque. The corollary of this situation is that antioxidants must be a benign force to protect the population from the modern scourge of heart disease. In fact, recent evidence from intervention studies with large doses of the antioxidant vitamins and other antioxidants in foods has been very disappointing. Here, the background for the belief that antioxidants ought to be beneficial is examined and an attempt made to explain why the results of these intervention studies have been unsuccessful. It is agreed that a diet rich in fruit and vegetables is protective for both CVD and cancer, but the explanation for this effect may not necessarily lie with the presence of antioxidants. PMID- 18412996 TI - New and emerging risk factors for CVD. AB - Morphological and immunocytochemical studies have elucidated the complex processes involved in atherogenesis. The notion of plaque instability has emerged from this work and underscored the importance of inflammation in determining clinical complications associated with atherosclerosis, such as acute coronary syndrome. Cells of the immune system have been detected within atherosclerotic lesions and auto-antibodies directed against modified LDL and heat-shock proteins have been identified in the blood of individuals with atherosclerosis. The use of risk 'engines', e.g. the Framingham coronary risk score, has facilitated the identification of individuals at high risk, but the constituent classical risk factors used in these algorithms do not adequately differentiate individuals at moderate risk. As age is a major component of the equations used in these algorithms they are not particularly useful in young adults, and their applicability to non-Caucasian populations has been questioned. Biomarkers of early disease and plaque instability have therefore both been sought. Although some of these markers have been shown individually to be associated with a significant hazard ratio, no substantial improvement in discrimination has been demonstrated when they are incorporated into a risk 'engine'. The latter has generally been assessed by receiver operator characteristic curve analysis, although this approach has been criticised. Other modalities, including imaging and functional assessments of vascular function, are now being developed for clinical use. PMID- 18412997 TI - Homocysteine, B-vitamins and CVD. AB - There is considerable interest in plasma homocysteine (tHcy) as a CVD risk factor. Although the secondary prevention trials published to date have been inconclusive in confirming a benefit of tHcy-lowering treatment with B-vitamins on CVD events generally, such studies are widely recognised to have been insufficiently powered to detect a significant effect for the predicted magnitude of association between tHcy and heart disease risk, and therefore cannot be interpreted as evidence that no relationship exists. In fact, a recent meta analysis of clinical trials has confirmed that folic acid supplementation reduces the risk of stroke, particularly in individuals without a history of stroke. Evidence supporting a causal relationship between elevated tHcy and heart disease also comes from genetic studies. The most important genetic determinant of tHcy in the general population is the common C677T variant in methylenetetrahydrofolate reductase (MTHFR) that results in higher tHcy. Individuals with the homozygous mutant (TT) genotype have a significantly higher (14-21%) risk of heart disease. Plasma tHcy is very responsive to intervention with the B-vitamins required for its metabolism, in particular folic acid, and to a lesser extent vitamins B12 and B6. Thus, although primarily aimed at reducing neural-tube defects, folic acid fortification may have an important role in the primary prevention of CVD via tHcy lowering. Besides folate, riboflavin is required as a cofactor for MTHFR and enhanced riboflavin status results in a marked lowering in tHcy specifically in individuals with the TT genotype, presumably by neutralising the variant form of the enzyme. About 10% of the UK and Irish populations have the TT genotype. In the present paper the potential role of folate and related B-vitamins in the primary prevention of CVD and the implications for nutrition policy are explored. PMID- 18412998 TI - Food for thought: the role of dietary flavonoids in enhancing human memory, learning and neuro-cognitive performance. AB - Emerging evidence suggests that dietary-derived flavonoids have the potential to improve human memory and neuro-cognitive performance via their ability to protect vulnerable neurons, enhance existing neuronal function and stimulate neuronal regeneration. Long-term potentiation (LTP) is widely considered to be one of the major mechanisms underlying memory acquisition, consolidation and storage in the brain and is known to be controlled at the molecular level by the activation of a number of neuronal signalling pathways. These pathways include the phosphatidylinositol-3 kinase/protein kinase B/Akt (Akt), protein kinase C, protein kinase A, Ca-calmodulin kinase and mitogen-activated protein kinase pathways. Growing evidence suggests that flavonoids exert effects on LTP, and consequently memory and cognitive performance, through their interactions with these signalling pathways. Of particular interest is the ability of flavonoids to activate the extracellular signal-regulated kinase and the Akt signalling pathways leading to the activation of the cAMP-response element-binding protein, a transcription factor responsible for increasing the expression of a number of neurotrophins important in LTP and long-term memory. One such neurotrophin is brain-derived neurotrophic factor, which is known to be crucial in controlling synapse growth, in promoting an increase in dendritic spine density and in enhancing synaptic receptor density. The present review explores the potential of flavonoids and their metabolite forms to promote memory and learning through their interactions with neuronal signalling pathways pivotal in controlling LTP and memory in human subjects. PMID- 18413000 TI - Neurodegeneration and the structure of time: clinical evidence for philosophical reasoning. PMID- 18412999 TI - A multi-centre prospective case-control study of campylobacter infection in persons aged 5 years and older in Australia. PMID- 18413001 TI - Innate immune response mechanisms in the intestinal epithelium: potential roles for mast cells and goblet cells in the expulsion of adult Trichinella spiralis. AB - SUMMARYGastrointestinal infection with the nematode Trichinella spiralis is accompanied by a rapid and reversible expansion of the mucosal mast cell and goblet cell populations in the intestinal epithelium, which is associated with the release of their mediators into the gut lumen. Both goblet cell and mast cell hyperplasia are highly dependent on mucosal T-cells and augmented by the cytokines IL-4 and IL-13. However, the contribution of both mast and goblet cells, and the mediators they produce, to the expulsion of the adults of T. spiralis is only beginning to be elucidated through studies predominantly employing T. spiralis-mouse models. In the present article, we review the factors proposed to control T. spiralis-induced mucosal mast cell (MMC) and goblet cell differentiation in the small intestine, and focus on some key MMC and goblet cell effector molecules which may contribute to the expulsion of adult worms and/or inhibition of larval development. PMID- 18413002 TI - Detection of Kobe-type Babesia microti associated with Japanese human babesiosis in field rodents in central Taiwan and southeastern mainland China. AB - Field rodent surveys for Babesia infection were performed from 2002 to 2005 in the vicinities of human babesiosis occurrences in Taiwan and mainland China. Babesia microti was identified by microscopical examination and/or PCR in 1 Rattus coxinga and 1 Crocidura horsfieldii in central Taiwan and in 13 Niviventer confucianus and 1 Apodemus agrarius in Zhejiang and Fujian Provinces of southeastern China. Of 15 B. microti samples detected by PCR, all except 1 were shown to be the Kobe-type, the aetiological small subunit rRNA gene-type of the first Japanese patient; the exception was also a Kobe-related type. The Kobe-type had been found in rodents only in a few places including the human infection occurrence place in Japan. The internal transcribed spacer 1 to 2 sequences of the Taiwanese and Chinese Kobe-types were very similar to each other but considerably different (approx. 94% pairwise identities) from that of the Japanese Kobe-type. A Taiwanese Kobe-type strain was serologically differentiated from the Kobe strain originating from the Japanese first patient. The distribution of the Kobe-type in the vicinities of human babesiosis occurrences in Taiwan and China as well as in Japan is suggestive of involvement of the Kobe type in Asian human babesiosis. PMID- 18413003 TI - Follow-up of antibody avidity in BALB/c mice infected with Toxocara canis. AB - In human Toxocara canis infection, an association has been shown between high IgG avidity in the chronic phase and low IgG avidity in recently acquired toxocarosis. The evolution of the antibody response in terms of avidity has been carried out through a T. canis infection in BALB/c mice. Infection with T. canis embryonated eggs (EE) was carried out with single doses (SD) of 6, 12, 50, 100, 200 or 1000 EE/mouse and with multiple doses (MD) of 200 and 1000 EE. Specific antibodies against T. canis (IgM+G, IgG, IgG1 and IgM) were detected by ELISA and Western Blot (WB) techniques in the presence and absence of urea. With the ELISA method, an increase in the avidity index (AI) of around 50% was detected from days 40-80 p.i. to the end of the study, with all the doses studied. The WB method showed the presence of high avidity antibodies bound to 100 kDa and 75 kDa T. canis proteins in all the cases when the IgM+G and the IgG1 antibodies were investigated. Antibodies of variable avidity were observed in those sera that recognized the group of low molecular weight proteins, between 37 kDa and 25 kDa. PMID- 18413005 TI - Multiplex PCR for differential diagnosis of emerging typhoidal pathogens directly from blood samples. AB - Classically Salmonella enterica serovar Typhi (S. Typhi) is associated with typhoid, a major health problem in developing countries. However, in recent years S. Paratyphi A and Vi-negative variants of S. Typhi have emerged rapidly. We have developed a nested multiplex PCR targeting five different genes for differential diagnosis of typhoidal pathogens which has been optimized to be directly applicable on clinical blood samples. Of 42 multiplex PCR-positive blood samples, 26, nine, and two were Vi-positive S. Typhi, Vi-negative S. Typhi and S. Paratyphi A, respectively, and five patients were found to have mixed infection. Seventeen patients grew Salmonella from blood culture and the remaining 25 were positive in the Salmonella-specific PCR. Tests with several common pathogens confirmed the specificity of the assay. We conclude that the proposed multiplex PCR is rapid, sensitive and specific for the diagnosis of typhoidal pathogens directly from blood samples. PMID- 18413004 TI - Socioeconomic disparities in the seroprevalence of cytomegalovirus infection in the US population: NHANES III. AB - There is a strong relationship between socioeconomic status (SES) and health outcomes in the United States, although the mechanisms are poorly understood. Increasing evidence points to links between lifelong exposure to infectious disease and subsequent chronic disease. Exposure and susceptibility to infections may be one way SES affects long-term health, although little population-based research to date has examined social patterning of infections in the United States. This paper tests the relationship between income, education, race/ethnicity and seroprevalence of cytomegalovirus (CMV) infection at different ages in a representative sample of the US population, and tests potential mediators for these relationships. The study finds significant racial and socioeconomic disparities in CMV seroprevalence beginning at early ages and persisting into middle age. Potential exposures do not explain the relationship between SES and CMV positivity. Because reactivation of latent CMV infections may contribute to chronic disease and immune decline later in life, future research should determine the exposure or susceptibility pathways responsible for these disparities in the prevalence of CMV infection. PMID- 18413006 TI - Hepatocellular integrity in patients requiring parenteral nutrition: comparison of structured MCT/LCT vs. a standard MCT/LCT emulsion and a LCT emulsion. AB - BACKGROUND AND OBJECTIVE: The aetiology of parenteral nutrition-associated hepatic injury remains unresolved. The aim of the study was to evaluate the effects of structured triglycerides in parenteral nutrition compared either to a physical medium-chain triglycerides (MCT)/long-chain triglcerides (LCT) mixture or to a LCT emulsion on hepatic integrity. METHODS: In a randomized, double blinded trial, we studied 45 patients undergoing abdominal surgery, who were expected to receive parenteral nutrition for 5 days. Patients were allocated to one of three nutrition regimens: Group A (n = 15) received structured triglycerides, Group B (n = 15) a MCT/LCT and Group C (n = 15) a LCT lipid emulsion. Before the start of parenteral nutrition (T0), 24 h (T1), 48 h (T2), 72 h (T3) and 120 h (T4) after start of infusion the following parameters were measured: Alpha-glutathione S-transferase (alpha-GST), alanine aminotransferase (ALT), aspartate aminotransferase (AST), glucose and serum triglycerides. RESULTS: At T3 and T4, alpha-GST levels were significantly higher in Group B (T3: 9.4 +/- 9.9; T4: 14.6 +/- 19.5 microg L-1) and Group C (T3: 14.2 +/- 20.8; T4: 22.4 +/- 39.3 microg L-1) compared with the patients receiving structured triglycerides (T3: 1.9 +/- 1.8; T4: 3.2 +/- 2.7 microg L-1). Whereas the mean alpha-GST-levels in structured triglycerides group always remained in the normal range, this was not the case in both other groups at T3 and T4. There were no significant differences concerning ALT, AST and glucose levels. At T3 and T4, triglyceride levels were significantly lower in Group A than in Groups B and C. CONCLUSIONS: Hepatic integrity was well retained with the administration of structured triglycerides, whereas both MCT/LCT emulsion and LCT emulsion caused subclinical hepatic injury. PMID- 18413007 TI - Acute normovolaemic haemodilution with a novel hydroxyethyl starch (130/0.4) reduces focal cerebral ischaemic injury in rats. AB - BACKGROUND AND OBJECTIVES: This study investigated the neuroprotective effect of acute normovolaemic haemodilution with a novel hydroxyethyl starch (130/0.4, HES) on focal cerebral ischaemia-reperfusion injury and determined optimum degree for haemodilution in the rat. METHODS: Male Sprague-Dawley rats were randomly divided into three groups (n = 10 each): hydroxyethyl starch, saline and controls. Animals in the hydroxyethyl starch and saline groups were haemodiluted until haematocrit decreased to 30% of baseline. In another experiment male Sprague Dawley rats were randomly assigned to four groups (n = 10 each): HES30, HES25 and HES20 and controls. Rats in the HES30, HES25 and HES20 groups were haemodiluted with hydroxyethyl starch until the haematocrit decreased to 30%, 25% and 20%, respectively. At 15 min after haemodilution, all the rats were subjected to a right middle cerebral artery occlusion for 120 min. Neurologic deficit scores and infarct volumes were assessed 24 h after reperfusion. RESULTS: Haemodilution with hydroxyethyl starch improved neurologic outcome (P < 0.01) and reduced infarct volume (P < 0.01) compared to saline and controls. Haemodilution with saline augmented infarct volume (P < 0.05 vs. controls). Neurologic deficit scores in HES30 and HES25 groups were significantly lower compared to controls (P < 0.05) while infarct volumes in controls and HES20 were significantly larger compared to HES30 and HES25 (P < 0.01). CONCLUSIONS: This study demonstrates that acute normovolaemic haemodilution with a novel hydroxyethyl starch (130/0.4) reduces infarct volume and improves neurological outcome after focal cerebral ischaemia in a rat stroke model. Neuroprotection by haemodilution was highest at haematocrit dilution between 25% and 30%, while ischaemic injury was exacerbates when haematocrit was diluted to 20%. PMID- 18413009 TI - Education and training in anaesthesia--revised guidelines by the European Board of Anaesthesiology, Reanimation and Intensive Care. AB - The expansion of the European Union during the last 2 yr has resulted in the need for a revision of existing guidelines to further harmonize education and training in the specialty of anaesthesiology throughout the European Union. Although each individual country is responsible for its own training and certification, these guidelines are intended to reflect minimum criteria for specialist training. It is the opinion of the Section and Board of Anaesthesia that specialist training will need to be for a minimum of 5 yr. PMID- 18413008 TI - Arterial blood gas derangement and level of comorbidity are not predictors of long-term mortality of COPD patients treated with mechanical ventilation. AB - BACKGROUND AND OBJECTIVE: Limited and inconsistent data exist on simple, readily available predictors of long-term mortality of critically ill chronic obstructive pulmonary disease patients requiring invasive mechanical ventilation. We therefore examined the influence of arterial blood gas derangement and burden of comorbidities on 90-day and 1-yr mortality of chronic obstructive pulmonary disease patients treated with invasive mechanical ventilation. METHODS: We identified all chronic obstructive pulmonary disease patients (n = 230) treated with invasive mechanical ventilation between 1994 and 2004 at a Danish primary level hospital. Data on arterial blood gas specimens and comorbidity were obtained from medical records and Hospital Discharge Registries. We used Cox's regression analysis to estimate mortality ratios according to arterial blood gas values and level of comorbidity. RESULTS: Ninety-day and 1-yr mortality among chronic obstructive pulmonary disease patients requiring invasive mechanical ventilation was 30.8% and 40.5%, respectively. All 90-day and 1-yr mortality ratios according to arterial blood gas values were close to one and one was included in all 95% CI. Among patients with a high level of comorbidity 90-day mortality ratio was 1.3 (95% CI: 0.6-2.7) when compared with patients without comorbidity. The corresponding 1-yr mortality ratio was 1.4 (95% CI: 0.7-2.9). CONCLUSION: Chronic obstructive pulmonary disease patients treated with invasive mechanical ventilation have substantial long-term mortality. Neither the levels of arterial blood gas values measured immediately before invasive mechanical ventilation was initiated nor the burden of comorbidity were strong determinants of long-term mortality among these patients. PMID- 18413010 TI - Regional anaesthesia for Caesarean section in an ankylosing spondylitic patient with twin pregnancy. PMID- 18413011 TI - The Public Health Nutrition workforce and its future challenges: the US experience. AB - OBJECTIVES: To describe the US public health nutrition workforce and its future social, biological and fiscal challenges. DESIGN: Literature review primarily for the four workforce surveys conducted since 1985 by the Association of State and Territorial Public Health Nutrition Directors. SETTING: The United States. SUBJECTS: Nutrition personnel working in governmental health agencies. The 1985 and 1987 subjects were personnel in full-time budgeted positions employed in governmental health agencies providing predominantly population-based services. In 1994 and 1999 subjects were both full-time and part-time, employed in or funded by governmental health agencies, and provided both direct-care and population-based services. RESULTS: The workforce primarily focuses on direct care services for pregnant and breast-feeding women, infants and children. The US Department of Agriculture funds 81.7 % of full-time equivalent positions, primarily through the WIC Program (Special Supplemental Nutrition Program for Women, Infants, and Children). Of those personnel working in WIC, 45 % have at least 10 years of experience compared to over 65 % of the non-WIC workforce. Continuing education needs of the WIC and non-WIC workforces differ. The workforce is increasingly more racially/ethnically diverse and with 18.2 % speaking Spanish as a second language. CONCLUSIONS: The future workforce will need to focus on increasing its diversity and cultural competence, and likely will need to address retirement within leadership positions. Little is known about the workforce's capacity to address the needs of the elderly, emergency preparedness and behavioural interventions. Fiscal challenges will require evidence-based practice demonstrating both costs and impact. Little is known about the broader public health nutrition workforce beyond governmental health agencies. PMID- 18413012 TI - Nutritional assessment interpretation on 22,007 Spanish community-dwelling elders through the Mini Nutritional Assessment test. AB - OBJECTIVES: To evaluate the prevalence of undernutrition among community-dwelling elderly people in Spain using the Mini Nutritional Assessment (MNA) and to analyse its distribution according to age, gender and residence region. DESIGN: Cross-sectional study assessing the nutritional status of elderly persons through both the short form and the full version of the MNA test. SETTING: Pharmacy offices across the whole country (Spain) were enrolled to recruit participants. SUBJECTS: A total of 22 007 participants (8014 men and 13 993 women), aged >or=65 years, assessed during the last two months of 2005. RESULTS: According to MNA screening, 4.3 % subjects were classified as undernourished (MNA score <17) and 25.4 % were at risk of undernutrition (MNA score >or=17 to 17 mm and serum oestradiol exceeded 200 (pg/ml). There was a non-significant shortening of induction cycles in group I. Oestradiol and LH concentrations were higher in group I compared with group II. Endometrial thickness, serum progesterone and clinical pregnancy rate were significantly higher in group I (8.9 +/- 1.4 mm versus 7.5 +/- 1.3 mm, P < 0.001; 13.3 +/- 3.1 ng/ml versus 9.3 +/- 2.0 ng/ml, P < 0.01; 36.7% versus 13.6%, P < 0.01, respectively). It is concluded that adding C. racemosa rhizome dry extract to clomiphene citrate induction can improve the pregnancy rate and cycle outcomes in these couples. PMID- 18413070 TI - Impact of assisted hatching on fresh and frozen-thawed embryo transfer cycles: a prospective, randomized study. AB - The aim of this study was to determine if assisted hatching (AH) could improve the rates of pregnancy and implantation for both fresh and frozen-thawed embryo transfer cycles. A total of 760 fresh embryo transfer cycles and 200 frozen thawed embryo transfer cycles were randomly assigned to either the treatment group (AH) or the control group (no AH). Zona thinning by laser was performed just before embryo transfer. In fresh embryo transfer cycles, the AH group and control group results were comparable. There were no significant differences in the rates of positive human chorionic gonadotrophin (HCG; 47.5 versus 48.8%), clinical pregnancy (42.4 versus 42.6%), or implantation (26.3 versus 25.2%) between the two groups. However, in frozen-thawed embryo transfer cycles, the rates of positive HCG (32.0 versus 17.0%), clinical pregnancy (25.0 versus 14.0%) and implantation (16.7 versus 7.3%) were significantly greater in the AH group than in the control group (P <: 0.05). The results of this investigation show that in the fresh embryo transfer cycles, laser-assisted hatching by zona thinning has no impact on the rates of positive HCG, clinical pregnancy and implantation, whereas in frozen-thawed cycles, assisted hatching by zona thinning significantly increases all three of these rates. PMID- 18413072 TI - Amniotic stem cell international. AB - Cells derived from the amniotic fluid for genetic diagnosis share many biological characteristics of stem cells. Recent studies at the single cell (clonal) level identified cells of the three embryonic germ layers in the amniotic fluid. It was therefore decided to establish an international non-profit alliance to create a repository of stem cells from the surplus cells remaining after completion of the cytogenetic analysis. PMID- 18413073 TI - Efficacy and safety of HMG versus r-FSH. PMID- 18413075 TI - Hyperreflexivity as a condition of mental disorder: a clinical and historical perspective. AB - Hyperreflexivity, understood as intensified self-consciousness in which subjects disengage from normal forms of involvement with nature and society, often considering themselves as objects of focal awareness, is proposed here as a condition of mental disorders, without which they would not exist. This thesis is argued from a dual perspective: clinical and historical. In the clinical perspective, it is shown that hyperreflexivity is not merely concomitant with mental disorders, but indeed has causal priority over them. Empirical evidence of a correlational, experimental and therapeutic nature, or deriving from cultural change, supports this claim of causal priority. In the historical perspective, it is shown that hyperreflexivity depends on certain historical-cultural circumstances that have prevailed since the Renaissance. These circumstances have to do with the emergence of the modern subject, displaced, autonomous and condemned to a hapless 'interior journey'. This means that mental disorders as such would not have existed prior to that era. Nor in the wake of the Renaissance would mental disorders automatically come into being, depending as they do on a reflexive, institutional clinical context, which would not emerge until practically the nineteenth century, but which would extend swiftly from then on. PMID- 18413076 TI - [Strategies of clinical hypothesis testing]. AB - This research examines whether Psychology students, when they test clinical hypotheses, follow either confirmatory or disconfirmatory reasoning strategies. Two hundred and six psychology students, divided in four groups, participated. One group received information about the probability that the hypothesis was correct by means of verbal labels, and another group, by means of numerical expressions. An additional group received the information that getting a precise diagnosis was clinically important. In a last group, diagnostic tests allowed them to increase certainty about the hypothesis. Results show a partial use of confirmatory strategies because, although participants did not seek confirming information, they indeed avoided collecting disconfirming information. When the information increased certainty about the hypothesis, confirmatory strategies became more likely. Neither the increase in the task importance nor the numerical expression of the likelihood that the hypothesis was correct seemed to affect the testing strategy used. PMID- 18413077 TI - [Personality disorders and psychopathy in men convicted for severe intimate partner violence]. AB - In this paper, a study of personality disorders and psychopathy in imprisoned male batterers is carried out. A sample of 76 batterers sentenced for a severe offence of violence against their intimate partner was selected. All the participants were assessed with the MCMI-II before beginning a treatment program in prison for gender violence. Likewise, all participants were assessed with the PCL-R in order to identify psychopathic symptoms. According to the results, 86.8% of the sample showed at least one personality disorder. The most prevalent one was the Obsessive-compulsive Personality Disorder (57.8% of cases), followed by the Dependent Personality Disorder (34.2% of cases) and Paranoid Personality Disorder (25%). Regarding psychopathy, the results of PCL-R showed that there were 11 people (14.4% of the sample) who met the criteria for psychopathy or probable psychopathy. Finally, implications of this study for clinical practice and future research in this field are commented upon. PMID- 18413078 TI - [Study of empathy in a Spanish sex offender sample]. AB - International research has emphasised that empathy is a relevant factor of sex offending. The main goal of this study is to explore, in a Spanish sex offender sample, the relationship between empathy deficits and sex crime for various types of victims. This study also analyses the positive effect that psychological treatment could have on empathy. For this purpose, 118 non-sex offenders were compared on an adapted version of Rape Empathy Measure to 39 treated rapists and 34 untreated rapists. Results show that the untreated rapists have lower empathy levels than non-sex offenders. Nevertheless, the treated rapists scored higher on empathy than the other two groups assessed, untreated rapists and non-sex offenders. Lastly, some suggestions are provided for future research in terms of the assessment of empathy and other factors related to sexual aggression. PMID- 18413079 TI - [Recidivism risk assessment in sex offenders]. AB - Violent behaviour risk assessment is one of the most relevant research areas in current Psychology of Crime. Various scales for violence risk assessment have recently been developed from research about crime careers and risk factors. One of these instruments is the Sexual Violence Risk Assessment-20 (SVR-20), translated and adapted to Spanish by the Group of Advanced Studies in Violence of the University of Barcelona. The goal of this study is to verify the predictive capacity of the SVR-20 to predict sexual violence recidivism in a Spanish sample of sexual offender inmates. The method used was a retrospective study based in 163 sexual offender files and a 4-year time lag. The data were analysed with the logistic regression technique. Of the sample, 79.9% non-recidivist individuals were correctly classified, and 70.8% recidivist individuals. The ROC curve obtained for the model shows a very good discriminant capacity for the SVR-20, with a 0.83 AUC value. The main conclusion of this study is that the Spanish adaptation of SVR-20 is a good instrument to predict the risk of sexual violence. PMID- 18413080 TI - [Alcohol drinking parties and consumption of alcohol and other drugs in youth]. AB - The main aim of this paper is to analyze the relationships of involvement in the "botellon" (meetings of youth in public places to drink, chat, etc.) with alcohol/other drug abuse and vandalism in young people. A school survey was performed in centres of Compulsory Secondary School (E.S.O.), high school, professional training and University education in Lugo, NW Spain. The sample (aged 12 to 30) was split into three groups according to alcohol use and botellon involvement; drug abuse, drug-related problems and vandalism were assessed in each group. Results indicate that regular botellon attendants display more problems with alcohol and other drugs, as well as higher involvement in vandalism; this relationship is stronger among young students. The increase in adolescent risk associated with the practice of the botellon is explained by peer group pressure to use drugs and the greater accessibility of alcohol/other drugs in these contexts. PMID- 18413081 TI - [Gender differences in alcohol dependence: personality variables, psychopathological profile and personality disorders]. AB - In this paper, gender differences in personality, psychopathology and personality disorders of alcohol-dependent patients are described. The sample consisted of 158 alcohol-dependent patients attending a psychiatric outpatient clinic (105 men and 55 women). All participants were assessed with various assessment tools related to personality (Impulsiveness Scale, Sensation Seeking Scale and STAI), psychopathology (SCL-90-R, BDI and Inadaptation Scale) and personality disorders (IPDE). There were no differences in personality variables, but the women had more anxiety and depressive symptoms and also more problems to adapt to everyday life than did the men. Personality disorders were not as prevalent as in the case of men, and the most frequent among women were obsessive-compulsive, dependent and histrionic personality disorders. Implications of this study for further research are commented on. PMID- 18413082 TI - [Efficient personality questionnaire for professional training]. AB - This study falls within the theoretical framework of the construct of the Efficient Personality. In this work, we present the study of validity and reliability of an original questionnaire made up of 60 items, applied to students from formative cycles of higher Professional Training. The questionnaire was reduced to 24 items, surpassing its initial .84 Cronbach alpha coefficient. Six first-order factors were obtained: Social Self-concept, Academic Self-concept, Solving Ability, Self-esteem, and Problem Coping. One second-order factor was obtained: Efficient Personality. The six-component factorial solution is similar both to the Spanish version for secondary school and to the Chilean versions for secondary and university populations, dividing the initial dimension of Self concept into Self-concept and Self-esteem. PMID- 18413083 TI - [Social support network and adjustment of minors in residential child care]. AB - In this study, the relationship between the social support network and the personal, social and academic adjustment of minors in residential care is analyzed. The sample consisted of 102 minors who lived in residential care and who completed the Social Support Questionnaire and the TAMAI. The results show that social support from the family, despite its being the greatest provider of support, does not affect the minors' adjustment. Neither does social support from peers affect their adjustment. However, social support from adults outside the family, and mainly their affection, seems to have a positive influence on the minors' adjustment, particularly at school and in social settings. These results are discussed in relation to their implications for the improvement of programs of residential child care. PMID- 18413084 TI - [Gender perspective in the context of Spanish academic psychology]. AB - The purpose of this study is to analyse the evolution of gender perspective in the context of the Spanish academic Psychology. The last three decades were analysed. In each decade, the study describes the groups and individuals who have contributed to the analysis of gender perspective in Psychology, the sponsoring institutions, and the most frequently used research strategies. Likewise, the study analyses the development of topics and areas in these three decades. The general trend shows that the interpretation of sex/gender has moved away from being considered an individual and internal factor to a more dynamic approach that analyses sex/gender through behavioural practices. PMID- 18413085 TI - [Tangibles as predictors of customer satisfaction in sports services]. AB - This study investigates the power of tangible dimensions of service quality to predict customer satisfaction. For this purpose, we statistically controlled the effects of dimensions of service quality that describe social interaction between employees and customers, both functionally and relationally. A field survey was conducted with the participation of 556 customers of a sports centre. The results showed that tangible dimensions of service quality predicted an additional and significant amount of customer satisfaction variance, beyond the effects of service quality of the social interaction. The article concludes with the discussion of the implications of these results. PMID- 18413086 TI - [Scale of expectations of control and responsibility: adaptation to the Spanish language and its relation to personal initiative]. AB - Personal initiative, understood as a proactive behaviour, self-initiated, persistent and pro-organization, is indispensable in contemporary organizations immersed in an environment of rapid and complex changes and innovations. In this work, one of the antecedents of personal initiative in the work setting is analyzed: the expectations or hopes of control and responsibility. We studied the adaptation to Spanish of the Scale of Expectations of Control and Responsibility of Frese and colleagues, using two cross-sectional samples (n=414 and 396, respectively). We performed exploratory and confirmatory factorial analyses that positively replicate the factor structure of the original scale. The relation of the expectations of control with personal initiative is confirmed, as suggested in the general model of antecedents and consequences of personal initiative. Likewise, the study confirms that control can be considered an antecedent of expectations of control and responsibility. PMID- 18413087 TI - [Motivational climate and coaches' communication style predict young soccer players' commitment]. AB - The purpose of this research was to study the role of coaches' communication style and coach-created motivational climate in young soccer players' enjoyment and commitment. Four hundred and fifteen young soccer players of high competitive level in the age range from 14 to 16 completed the following questionnaires: a) coach-induced perceived motivational climate (PMCSQ-2), b) coaches' behaviour perception (CBAS-PBS), and c) sport commitment (SCQ). Results showed that coach created motivational climate correlated highly with the perception of coaches' communication style. Moreover, coach-created motivational climate and communication style significantly determines players' sport commitment and enjoyment. Discussion focuses on the importance of seeking and training credible coaches that favours athletes' commitment. PMID- 18413088 TI - [Achievement and social goals, persistence-effort, and intentions of sport practice in physical education students]. AB - The present study examined the relationship between achievement and social goals, and explored how both types of goals can affect secondary school students' persistence/effort in Physical Education classes, as well as their intention to participate in sport activities after finishing secondary school. A sample of 350 students (aged 15-17) from two high schools in Asturias participated in the study. Multiple regression analysis showed: a) persistence/effort in physical education classes was better explained by social responsibility goals, followed by mastery-approach goals and mastery-avoidance goals; b) achievement goals are the only predictors of young people's intention of practicing sport in the future. Differences were found in these results according to gender and course. These findings help us better understand the processes involved in Physical Education. Finally, implications to improve classes and to achieve young people's more active and higher level of implication in sports are discussed. PMID- 18413089 TI - [Quality of life in transplant patients, compared to other stressful health situations in pulmonary patients]. AB - Transplantation may imply severe biopsychosocial impairments. In order to know the quality of life of patients one year after transplantation, 58 subjects were compared to three different groups of patients (stabilized and acute COPD patients, and lung cancer patients in a surgery unit). Patients filled in two questionnaires: EORTC QLQ-C30 (quality of life) and HAD (anxiety and depression). The quality of life dimensions with inter-group differences were physical, role, emotional and cognitive functioning, global health status, and a number of symptoms (fatigue, dyspnea, insomnia and appetite loss). There were differences in depression, and but not in anxiety. Transplant and surgical patients showed better quality of life and affective status than chronic pulmonary patients. Discriminant analysis showed that the transplant group was the best described group. We conclude that patients, one year after transplantation, show similar quality of life as asymptomatic hospitalised patients, somewhat better than chronic patients in a stabilized stage of the disease, and much better than severe chronic patients. PMID- 18413090 TI - [Impact of low-level expression dermatological diseases on the quality of life as a function of its level of specificity]. AB - The impact of dermatological diseases on the quality of life is relatively well established, although some results do not support this relationship. Whereas a few studies indicate a decrease in various indicators of quality of life in conditions of disease, in other studies, this effect has not been observed. These differences can be explained by the use of different concepts of quality of life and/or the comparison of different severity levels of the diseases. The purpose of this paper is to analyze the impact of low-level expression dermatological diseases on various concepts of quality of life. Two hundred-ninety-nine participants completed a questionnaire about symptoms, psychological and psychosocial impact, and general and specific satisfaction. Results show that minor dermatological diseases affect health-related quality of life, but not quality of life considered as well-being. PMID- 18413091 TI - [Effects of the iodine deficiency on intellectual variables among children]. AB - An association between severe iodine deficiency and poor mental development has been found in many studies. We examined the relationship between moderate or mild iodine deficiency and intellectual capacity in order to determine whether problems common to severe iodine deficiency (including mental retardation) also emerge in a more subtle form. We also wished to know whether the classic methodology (comparing iodine-deficient zones with nondeficient zones) is the most adequate, and propose to combine this grouping by zones with urinary iodine presented by individuals in each zone. We measured IQ, manipulative and verbal capacity, attention, visual motor ability and disruptive behaviour, variables that have barely been studied in this kind of investigations. The sample comprised 760 schoolchildren from the province of Jaen (southern Spain). Our results show that children with low levels of iodine intake and with urinary iodine concentration lower than 100 microg/litre had a lower IQ and displayed more disruptive behaviour than children with high levels of the criteria. The other variables were not associated with iodine deficiency. PMID- 18413092 TI - [Effects of extinction in multiple contexts on the renewal of associative tolerance to ethanol]. AB - The Pavlovian analysis of drug tolerance describes how the contexts of drug administration participate in the elicitation of compensatory conditioned responses that are, at least partly, responsible for tolerance. Findings indicate that if an association is acquired in one context and extinguished in another one, when exposing the subjects to the acquisition context, renewal of the conditioned response will take place. Ambiguous evidence exists about whether or not this effect diminishes when extinguishing the association in multiple contexts. This investigation evaluates the existence of the phenomenon of renewal of tolerance to ethanol, and whether it diminishes by the extinction in multiple contexts. Evidence of renewal of tolerance to ethanol in rats was found, but no diminution was observed when extinguishing in multiple contexts. PMID- 18413093 TI - [SA-45: a brief form of the SCL-90]. AB - The Symptom Assessment-45 Questionnaire (SA-45) is a 45-item self-report instrument of psychiatric symptomatology derived from the original SCL-90. The SA 45 consists of nine 5-item scales assessing each of the same symptom domains as its parent instrument with no item overlap across domains. This paper provides preliminary validation of the Spanish version of the questionnaire in an undergraduate sample. Exploratory and confirmatory factor analyses supported a 9 factor structure, which corresponds to the nine proposed scales. Normative data, reliability (internal consistency) and validity were also examined, finding support for sound psychometric properties. PMID- 18413094 TI - Randomization tests for ABAB designs: comparing-data-division- specific and common distributions. AB - Monte Carlo simulations were used to generate data for ABAB designs of different lengths. The points of change in phase are randomly determined before gathering behaviour measurements, which allows the use of a randomization test as an analytic technique. Data simulation and analysis can be based either on data division-specific or on common distributions. Following one method or another affects the results obtained after the randomization test has been applied. Therefore, the goal of the study was to examine these effects in more detail. The discrepancies in these approaches are obvious when data with zero treatment effect are considered and such approaches have implications for statistical power studies. Data-division-specific distributions provide more detailed information about the performance of the statistical technique. PMID- 18413095 TI - [Factorial confirmatory models of Ryff's scales in a sample of elderly people]. AB - Several recent works have analysed the factorial structure of well-being measures. The aim of our study is to analyse the factorial structure of a widely used well-being scale, Ryff's Scales of Psychological Well-being, but in a specific subpopulation of the Spanish population, the elderly. For this particular subpopulation, the construct of well-being has been employed in most theoretical models that explain quality of life, and its role is therefore pivotal. The sample comprised 169 elderly people (65 years or more), sampled within the Valencian Community. The 54-item version of Ryff's scales was used. An item parcelling process was analytically employed before confirmatory factor analyses, allowing a total of 18 well-being indicators. Confirmatory factor analyses were specified and tested, including all theoretical and empirical solutions found in the literature, either in the general population or in specific populations of different cultural contexts. Goodness-of-fit results were similar to the ones found in the literature. Best solutions were a six-factor model with correlated factors, as defended by the authors, and a five-factor correlated solution, collapsing environmental mastery and self-acceptance into a single factor. PMID- 18413096 TI - [Determining the most unfavourable variance to calculate the Measurement Scale Imprecision Factor, and extension to other types of sampling methods]. AB - The precision of estimates must be adequately reported in survey research, where ordinal and interval measurement scales are commonly used. Regarding mean estimate, absolute and relative errors exist as a function of the measurement scales. This manuscript discusses some assumptions underlying the development of the Measurement Scale Imprecision Factor--MSIF--, a tool to assess the degree of imprecision of estimates, regardless of the scale rank considered. Specifically, we propose a new method for determining the most unfavourable variance, which is consistent with the normal distribution assumption, unlike the original assumption based on the bimodal distribution. This method reduces the value of the most unfavourable variance, which is easily computed using the cumulative normal standard distribution function. In addition, we show the relationship between MSIF and other types of probabilistic sampling methods, such as stratified and cluster sampling. PMID- 18413097 TI - [Validity evidence of the Health-Related Quality of Life for Drug Abusers Test based on the Biaxial Model of Addiction]. AB - The aim of this work is to show evidence of the validity of the Health-Related Quality of Life for Drug Abusers Test (HRQoLDA Test). This test was developed to measure specific HRQoL for drugs abusers, within the theoretical addiction framework of the biaxial model. The sample comprised 138 patients diagnosed with opiate drug dependence. In this study, the following constructs and variables of the biaxial model were measured: severity of dependence, physical health status, psychological adjustment and substance consumption. Results indicate that the HRQoLDA Test scores are related to dependency and consumption-related problems. Multiple regression analysis reveals that HRQoL can be predicted from drug dependence, physical health status and psychological adjustment. These results contribute empirical evidence of the theoretical relationships established between HRQoL and the biaxial model, and they support the interpretation of the HRQoLDA Test to measure HRQoL in drug abusers, thus providing a test to measure this specific construct in this population. PMID- 18413098 TI - Spanish normative data of the Zuckerman-Kuhlman Personality Questionnaire in a general population sample. AB - The Zuckerman-Kuhlman Personality Questionnaire (ZKPQ), a tool designed to measure the alternative five-factor model of personality, was translated and adapted into the Spanish language. To date, there appears to be no normative data for the ZKPQ in general population samples in any culture. The aim of this study was to obtain Spanish normative data for the scales and subscales in a community sample of the general population. The questionnaire was administered to 1,678 participants of both genders (55.8 % females) with ages ranging from 18 to 93 years. The magnitude of the sample, as well as an adequate representativity of both genders and the wide age range being considered, provides a satisfactory description of each scale and subscale of this instrument. The normative data obtained showed a normal distribution of the scales. The norms (means and standard deviations) are presented by gender and five age ranges and are adjusted according to the level of education attained. Gender differences are in accordance with data obtained with instruments measuring similar personality traits. This is the first time the normative data of the ZKPQ is reported in a general community sample in any culture. PMID- 18413099 TI - [Evaluation of three sensitivity measures in magnitude estimation tasks]. AB - Taking into account the studies about the measure of sensitivity in magnitude estimation tasks, we analyze the three most common measures used in this topic: Pearson's product-moment correlation between the logarithm of the stimulus and the logarithm of the response (R), the exponent of Stevens' power function (K), and the measure "M" proposed by Garriga-Trillo. Using a sample of participants greater than usual in psychophysical studies (180 participants), we designed an experiment with two sets of stimuli with different stimulus ranges. In each of these sets, we used two kinds of stimuli (line segments and squares). Our conclusions were: (1) we rejected the use of K as a sensitivity measure because the results provided by this index were the opposite of those expected when we compared the two stimulus ranges. (2) We also rejected the use of M because this measure is a linear transformation of Kendall's coefficient of concordance. (3) Lastly, we suggest the mathematical transformation proposed by Fisher to achieve a normal distribution, and recommend this transformation as the best sensitivity measure. PMID- 18413100 TI - Ethanol-induced flushing with topical pimecrolimus use. AB - Topical calcineurin inhibitor use has been implicated in a flushing reaction associated with ethanol ingestion. This adverse effect has mostly been described as occurring with the application of tacrolimus ointment, and one report associated it with the use of pimecrolimus cream. We present three cases of ethanol-induced flushing in patients who used topical pimecrolimus. PMID- 18413101 TI - Contact allergy to p-tert-butylphenol formaldehyde resin and zinc diethyldithiocarbamate in a wet suit. AB - Rubber materials are common causes of contact dermatitis. Neoprene is a special synthetic rubber used in many products (eg, wet suits, elastic supports, gloves, shoes, and orthopedic devices). A 31-year-old man was admitted to our dermatoallergologic clinic with the development of a generalized itching erythematovesicular eruption. He reported that clinical manifestations occurred after he wore a neoprene wet suit that he was used to wearing for water sports. Although allergic contact dermatitis from a wet suit is not uncommon, it is usually due to thiourea derivatives whereas our patient presented with contact allergy to p-tert-butylphenol formaldehyde resin and zinc diethyldithiocarbamate. PMID- 18413102 TI - Contact allergic dermatitis from melamine formaldehyde resins in a patient with a negative patch-test reaction to formaldehyde. AB - Melamine paper is a basic material used in the furniture industry for home and office interiors. Contact allergic dermatitis from melamine formaldehyde resins (MFRs) should be considered in patients who work on melamine paper impregnation lines. We report a case of a 28-year-old female plywood worker who developed eczema on the dorsal side of her hands and wrists after 2 years of working on the melamine paper impregnation line. She had a relevant positive patch-test reaction to MFR, with a negative reaction to formaldehyde. Contact dermatitis due to MFR is not common, and it is usually related to products that are not fully cured or to close contact with intermediate products on the assembly line. Formaldehyde release from MFR can explain most of the positive responses. To our knowledge, this is the first report of MFR contact allergic dermatitis in a worker on a melamine paper impregnation line. PMID- 18413103 TI - Neomycin. PMID- 18413104 TI - Compositae. PMID- 18413107 TI - Grenz ray therapy in the new millennium: still a valid treatment option? AB - Grenz ray therapy has declined in the United States during the past several decades with respect to availability, use, and resident training. However, it remains a simple, affordable, and effective option for managing several refractory dermatoses, especially psoriasis, hand eczema, and allergic contact dermatitis. Though high-dose grenz irradiation has been associated with the development of nonmelanoma skin cancers, grenz ray therapy is considered a safe treatment modality when administered according to the guidelines recommended herein. We review the value of grenz irradiation in contemporary dermatology. PMID- 18413106 TI - Lanolin allergy: history, epidemiology, responsible allergens, and management. AB - Allergy to lanolin has been recognized by dermatologists for decades. This review summarizes the history, epidemiology, and allergenicity of lanolin and its derivatives. "The lanolin paradox" and the safety of pharmaceutical-grade lanolin products are also discussed. PMID- 18413108 TI - Patch-test reactions to topical anesthetics: retrospective analysis of cross sectional data, 2001 to 2004. AB - BACKGROUND: Allergy to topical anesthetics is not uncommon. The cross-reactivity among topical anesthetics and the screening value of benzocaine alone are not well understood. OBJECTIVES: The goals for this study were: (1) to evaluate the frequency and pattern of allergic patch-test reactions to topical anesthetics, using North American Contact Dermatitis Group (NACDG) data, and (2) to compare these results to allergen frequencies from other published studies. METHODS: The NACDG patch-tested 10,061 patients between 2001 and 2004. In this analysis patients were included who had positive patch-test reactions to one or more of the following: benzocaine, lidocaine, dibucaine, tetracaine, and prilocaine. RESULTS: Of patch-tested patients, 344 (3.4%) had an allergic reaction to at least one anesthetic. Of those, 320 (93.0%) had an allergic reaction to only one topical anesthetic. Overall, reactions to benzocaine (50.0%, 172 of 344) were most prevalent, followed by reactions to dibucaine (27.9%, 96 of 344); however, reactions to dibucaine were significantly more frequent in Canada than in the United States (relative risk [RR], 2.31; 95% confidence interval [CI], 1.67-3.20; p < .0001). Of patients reacting to more than one anesthetic, most (79%, 19 of 24) reacted to both an amide and an ester. CONCLUSIONS: Of the topical anesthetics tested, benzocaine was the most frequent allergen overall. Over 50% of allergic reactions to topical anesthetics in this study would have been missed had benzocaine been used as a single screening agent. Cross-reactivity patterns were not consistent with structural groups. PMID- 18413109 TI - Allergic contact dermatitis in children: the Ottawa hospital patch-testing clinic experience, 1996 to 2006. AB - BACKGROUND: Allergic contact dermatitis in children is a significant clinical problem. Patch testing is a diagnostic tool for the evaluation of patients with suspected allergic contact dermatitis. OBJECTIVES: To determine the frequency and relevance of positive patch-test results in children and to identify the most common allergens in children at our clinic. METHODS: Retrospective chart review of 100 children and adolescents, aged 4 to 18 years, who were patch-tested at the Ottawa Hospital patch-testing clinic between 1996 and 2006. The children were patch-tested with the North American Contact Dermatitis Group (NACDG) standard series, supplementary series if indicated, and their own products if available. RESULTS: Seventy percent of children had at least one positive patch-test reaction; 55.8% of positive patch-test reactions were relevant. The ratio of females to males was 62:38. The most common allergens were nickel sulfate (26%), cobalt (14%), fragrance mix (7%), neomycin (7%), colophony (6%), formaldehyde (4%), lanolin (4%), quaternium-15 (4%), and para-phenylenediamine (4%). Nickel co reacted with cobalt (68%) and palladium (100%). Of children tested, 41% had a history of atopic dermatitis. CONCLUSIONS: The frequency of positive and relevant allergens in children is similar to that in adults as compared with data from the NACDG 2001-2002 study period. Differences between the top 10 allergens in children and adults were seen. Nickel and cobalt were more common allergens in children, and colophony, lanolin, and para-phenylenediamine ranked in the top 10 allergens among children. PMID- 18413110 TI - Efficacy and patient perception of Grenz ray therapy in the treatment of dermatoses refractory to other medical therapy. AB - BACKGROUND: Grenz ray therapy (GRT) has been used to treat dermatoses refractory to other therapy. Patient's assessments to GRT alone have not been previously reported. OBJECTIVE: (1) To report the patterns of use of GRT at our center over a 10-year period and evaluate the efficacy of GRT in treating recalcitrant skin conditions, (2) to evaluate patient perceptions about GRT, and (3) to evaluate whether patients felt this form of treatment was worthwhile. METHOD: Cross sectional survey by mailed questionnaire to patients treated with GRT from 1990 to 2001. RESULTS: Of 351 patients treated with GRT from 1990 to 2001, 98 (28%) returned the questionnaire; 64% reported decreased severity or clearing of disease (p = .003), and 63% reported decreased or no discomfort (p = .006) 3 months following treatment. Overall, 54% said GRT was worthwhile, and 53% would choose it again; 40% reported mild side effects. Number of treatments (p = .2) or total dose (p = .25) were not significantly different among responders to GRT and nonresponders to GRT. In a subgroup of treated patients with a diagnosis of contact dermatitis (94% with hand dermatitis), 64% felt GRT was worthwhile and 77% indicated that they would choose this therapy again if needed. CONCLUSIONS: Many patients treated with GRT for recalcitrant dermatitis reported that this treatment was an effective therapy in decreasing the discomfort and severity of their skin condition. Overall, just more than half of treated patients believed GRT was a worthwhile therapy that they would use again. PMID- 18413111 TI - Determination of in vivo dose response and allergen-specific T cells in subjects contact-sensitized to squaric acid dibutyl ester. AB - BACKGROUND: Squaric acid dibutyl ester (SADBE) is a known contact sensitizer, but dose-response data are not defined. OBJECTIVE: To determine the relationship between sensitization dose and contact hypersensitivity (CHS) response to SADBE in human volunteers. The study also aimed to investigate whether SADBE-reactive blood T cells could be detected using ex vivo mature dendritic cells (DCs) as antigen-presenting cells. METHOD: Forty healthy volunteers were sensitized to either 12.5, 25, 50, or 250 microg of SADBE in a 48 microL volume. This was followed by elicitation 2 weeks later with five doses (0, 0.2, 2, 20, and 200 microg in 20 microL). An additional 10 subjects received the elicitation doses without prior sensitization. Blood samples obtained after sensitization were purified into T cells and mature DCs. RESULTS: A direct relationship between sensitization dose and in vivo CHS response was observed. The SADBE dose that effectively sensitized 50% of the population (ED50) was 22 microg/cm2. Significant SADBE-specific T-cell proliferation in vitro was not observed 2 weeks after sensitization but became evident after elicitation. CONCLUSION: This study establishes the in vivo dose-response characteristics of immune reactivity to SADBE and antigen-specific T-cell reactivity. PMID- 18413112 TI - Spa contact dermatitis. AB - Potassium monopersulfate (MPS) is widely used in spa and pool "shock" treatments, yet contact dermatitis associated with MPS has been rarely reported. A patient presented with a generalized scattered dermatitis from the neck down that worsened after spa use. Patch testing elicited a ++ reaction to ammonium persulfate. Contact with ammonium persulfate was ruled out; however, MPS, which can cross-react with ammonium persulfate, was found to be the active ingredient in the patient's spa shock treatments. The dermatitis cleared after the patient switched to a hydrogen peroxide-based shock treatment. PMID- 18413113 TI - Self-treatment with bleach by a Latino farmworker. AB - Skin disease is common among migrant Latino farmworkers. These workers rarely use formal health care services but commonly engage in self-treatment of their skin disease. We present a patient with dermatitis who self-treated with bleach. This patient illustrates a common practice that exacerbates skin disease and sheds light on social and cultural factors of which health care providers serving this community should be aware. PMID- 18413114 TI - Systemic contact dermatitis from propylene glycol. AB - A 39-year-old woman presented with pruritic eczematous plaques on her face, neck, and right hand that she had had for approximately 2 months, following an abrasive injury caused by the deployment of an airbag in a car accident. Results of patch testing were positive for several medicaments and propylene glycol (PG). The patient's condition cleared after discontinuation of all topical products containing PG and her other identified allergens, but she noted flares of her contact dermatitis following the ingestion of foods containing PG. A subset of patients will have a recurrence of dermatitis after the ingestion of a contact sensitizer. Recurrent dermatitis despite complete avoidance of identified topical allergens and a history of recurrent eczema at the patch-test site are clues to the diagnosis of systemic contact dermatitis. Even weak patch reactions to PG, if they persist to a day-7 reading, should be considered potentially relevant. Avoidance of dietary PG includes attention to labels on food and medication and the avoidance of certain foods in restaurants when ingredients cannot be verified. PMID- 18413117 TI - Improvement of the T.R.U.E. Test: launch of panel 3. PMID- 18413115 TI - Contact dermatitis from a prosthesis. AB - Patients wearing a prosthesis face a wide variety of medical problems. Skin complications have long been recognized, but their prevalence is still unknown. The most frequently reported disorders are allergic contact dermatitis (ACD), acroangiodermatitis, epidermoid cysts, epidermal hyperplasia, follicular hyperkeratosis, verrucous hyperplasia, bullous diseases, hyperhidrosis, infections, malignancies, and ulcerations. Contact dermatitis represents one third of the dermatoses in amputees wearing prostheses. All patients who are suspected of having ACD should be patch tested with standard allergen series as well as materials from the patient's own prosthesis, topical medicaments, moisturizers, and cosmetics. We report a patient with an ACD to mixed dialkyl thiourea present in the rubber parts of his below-the-knee prosthesis. Thiourea derivates are used as accelerators in the manufacture of chloroprene rubber and as fixatives in photography and photocopy paper. Allergy to thiourea is relatively uncommon; different studies have shown a prevalence of 0.7% up to 2.4% in patch-tested patients. Thiourea derivates are often the allergic sources in ACD involving high-grade rubber products made of neoprene such as diving suits, protective goggles, knee braces, and continuous positive airway pressure masks. They are also present in the rubber material of prostheses, as in the case of our patient. PMID- 18413118 TI - Successful patch testing despite concomitant low-dose prednisone use. PMID- 18413119 TI - [Characteristics and prognosis of patients with very long stay in an Intensive Care Unit]. AB - OBJECTIVE: To describe the characteristics and prognostic factors of elderly patients hospitalized for > or = 30 days in an Intensive Care Unit (ICU). DESIGN: Retrospective analysis of prospectively collected simple data over 6 consecutive years. SETTING: Polyvalent ICU of the Insular University Hospital in the Canary Islands (Spain). PATIENTS: Adult patients > or = 70 years who were hospitalized in the ICU for a period of 30 or more days. PRIMARY VARIABLES OF INTEREST: Demographic data, clinical diagnosis on ICU admission, Apache II, days of renal replacement therapy (RRT), days of mechanical ventilation and the outcome of the survivors one year later were collected. Mortality at one year of the surviving patients was studied. RESULTS: During the study period, 3,786 patients were admitted to the ICU. Of these, 853 (22.5%) patients were > or = 70 years old and only 42 (4.92%) of these patients remained in the ICU for > or = 30 days. We compared the latter with the > or = 70 year old patients whose stay in the ICU stay was < 30 days. No statistically significant differences in ICU mortality, Apache II, age, gender and the need for RRT were found. As independent variables associated with the long stay, the multivariate analysis showed only the days of mechanical ventilation (p < 0.05). The surviving patients (> or = 70 years old and whose stay in the ICU was > or = 30 days) were older and 21 (65.62%) were still alive one year later. CONCLUSIONS: ICU mortality rates in elderly patients with a stay < or > or = 30 days in the ICU were comparable. Survival at one year of the > or = 70 year-old patients whose long-term intensive care unit stay was > or = 30 days was high. These results are sufficient in our unit to justify prolonged ICU care for elderly patients. PMID- 18413120 TI - [Acute renal function deterioration in a Coronary Unit in Spain]. AB - OBJECTIVE: To assess the incidence and prognostic value of acute renal function deterioration (ARFD) in patients with acute heart disease. SETTING: Coronary Care Unit in a tertiary university hospital. DESIGN: Retrospective study. PARTICIPANTS: One hundred consecutive patients admitted during 2004. INTERVENTIONS: No randomized interventions were done. Diagnostic and therapeutic procedures were performed according to local protocols and current Clinical Practice Guidelines. PRIMARY VARIABLES: The primary aim of the study was to analyze the incidence of acute renal function deterioration and its effect in mortality during hospitalization. ARFD was defined as the increase of serum creatinine by 0.5 mg/dl and/or by 50% over baseline. RESULTS: Incidence of ARFD was 26%, with a mean increase of serum creatinine of 1.5 +/- 0.9 mg/dl. ARFD was significantly associated with age, background of hypertension and chronic kidney disease. Patients with ARFD had a more complicated course, longer hospitalizations, and received fewer catheterisms. Acute renal function deterioration was associated with higher mortality during hospitalization (33% versus 6%, p = 0.002). CONCLUSIONS: Acute renal function deterioration is frequent in patients with acute heart disease and its presence is linked with higher mortality. PMID- 18413121 TI - [Acridine orange staining method in the diagnosis of catheter-related bloodstream infections]. AB - OBJECTIVE: To study if the utility of acridine orange (AO) staining method on blood extracted through intravenous device (ID) is a reliable method to diagnose catheter-related bloodstream infection (CRB). DESIGN: Prospective and observational study. PATIENTS: Patients with central ID and clinical data consistent with CRB who gave their consent to participate. Patients having another infection site were excluded. INTERVENTION: At the moment of the clinical suspicion of CRB and before removing the ID, blood samples were extracted from peripheral veins and through the ID to be analyzed by AO staining. After extracting the samples, the catheter was removed and sent for microbiological analysis with Linares et al and Maki et al techniques. CRS was defined as development of the same microorganism in the tip of the catheter (endoluminal surface with > or = 10 (3) UFC/ml and/or extraluminal surface > or = 15 UFC/ml) and in the peripheral blood. VARIABLES OF INTEREST: Sensitivity, specificity, negative and positive and negative predictive values and positive likelihood ratios (LR) were calculated for the diagnosis of CRB. RESULTS: A total of 121 patients were studied and 4 were diagnosed with CRB: 2 infected with Staphylococcus aureus, 1 with Pseudomonas aeruginosa and 1 with Candida albicans. AO sensitivity was 87.5%, specificity 92.7% and the negative predictive value was 99.5%. Positive likelihood ratio was 12.04 and negative LR 0.13. CONCLUSIONS: Although the number of events does not allow for the estimation of the efficacy of AO to diagnose CRB, its high negative predictive value would make it possible to rule out this infectious complication with some degree of safety. PMID- 18413122 TI - [The neurocritical patient]. PMID- 18413123 TI - [Status epilepticus]. AB - Status epilepticus is a neurological emergency that requires prompt care. The diagnosis and treatment must be continuous from the first minutes to its resolution. The most frequent cause in epileptic patients is drug change or non compliance and in the non-epileptic patients are cerebrovascular diseases, head trauma, drug toxicity and metabolic disturbances. Status epilepticus can be convulsive or non-convulsive and continuous electroencephalographic monitoring is useful for diagnosis and to evaluate response to treatment. Benzodiazepines and phenytoin or fosphenytoin are first-line and second-line therapy. There is no agreement on third and fourth line therapy: phenobarbital, valproate, levetiracetam, propofol, midazolam, barbiturates and others. The prognosis of status epilepticus is related to etiology, age, type and duration of the status. Thus, drug treatment for status epilepticus should be started without delay. PMID- 18413124 TI - [Bird flu: what the intensivist must know]. AB - In the last century, humankind has faced 3 major pandemics of influenza virus infections. The first one occurred in 1918 and caused a significant amount of deaths. It was also capable of crossing over species barrier and affecting mammals, and most worrisome, humans. Since then several outbreaks have been reported in the Southeast of Asia. Many patients with the flu-like illness have a severe course and the patient develops pneumonia and in some cases multiorgan failure involving liver, kidneys, brain and lungs. Since the virus lacks regulatory control of genetic division it undergoes constant mutations leading to new subtypes and, sometimes, new strains. The only drugs that have shown some protection are oseltamivir and zanamivir. It is crucial to develop effective and non-expensive vaccines to prevent the virus spread and infection not only in humans but in birds too. PMID- 18413125 TI - [Management of thoracic aorta traumatism in 5 multiple traumatized patients]. AB - Thoracic aorta traumatism is a lesion associated to high morbidity-mortality at the site of the accident. In the 90's, treatment by placement of an endovascular stent was generalized with good results. In this work, we present a series of 5 clinical cases of patients admitted to the Intensive Care Unit of our hospital during the year 2006. Out of a total of 619 patients admitted in the same period, 121 (19.5%) were traumatic and 5 (4.13%) had thoracic aorta traumatisms. All of the patients were diagnosed by thoracic helical computed tomography on admission. Four patients had a pseudoaneurism of the aorta and underwent an operation. Three received endovascular treatment within the first 36 hours and the fourth on day 28 of admission. The fifth patient had a lesion of the intima and was not operated on. Posterior evolution was good in all the cases. PMID- 18413126 TI - [Recommendations in sedoanalgesia of the critical patient]. PMID- 18413127 TI - [Cerebral venous sinus thrombosis: an uncommon neurological emergency]. PMID- 18413129 TI - [False image of pneumothorax in simple X-ray]. PMID- 18413130 TI - Cellular pathogenesis in prion diseases. AB - Prion diseases are characterised by neuronal loss, vacuolation (spongiosis), reactive astrocytosis, microgliosis and in most cases by the accumulation in the central nervous system of the abnormal prion protein, named PrP(Sc). In this review on the "cellular pathogenesis in prion diseases", we have chosen to highlight the main mechanisms underlying the impact of PrP(C)/PrP(Sc) on neurons: the neuronal dysfunction, the neuronal cell death and its relation with PrP(Sc) accumulation, as well as the role of PrP(Sc) in the microglial and astrocytic reaction. PMID- 18413131 TI - Quantification of porcine circovirus type 2 (PCV-2) within- and between-pen transmission in pigs. AB - PCV-2 within- and between-pen transmission was quantified by estimating the daily transmission rate beta and the basic reproduction ratio (R(0)) using a stochastic SEIR (Susceptible, Exposed, Infectious, Removed) model fitted on experimental data. Within-pen transmission was quantified by using four groups of eight SPF (specific pathogen-free) pigs (four infected and four susceptible pigs having direct contact). Between-pen transmission was studied in two groups of 16 SPF pigs (eight infected and eight susceptible pigs having indirect contact (10 cm distance)). Pigs were monitored twice a week (blood samples) and were tested for PCV-2 antibodies (ELISA test) and viral genome load in sera (real-time PCR). Transmission parameters beta(within) and beta(between) were estimated using a maximum likelihood method and the duration of infectiousness, to compute R(0), was estimated with a parametric survival model. Different assumptions were made to determine the end of infectiousness (seroconversion, seroconversion and decline in viral genome load, permanent infectiousness). R(0[within]) (8.9 (5.1 15.4)) was greater when the end of infectiousness was assumed to be related to both seroconversion and a decline of PCV-2 genome load in sera (average duration of infectiousness = 32 days) compared with only seroconversion as the indicator of recovery (R(0[within]) = 5.5 (3.3-9.0)). Whatever the assumption, between-pen R(0) (0.58 (0.23-1.47)) was always significantly lower than within-pen R(0). Only beta(within) was sensitive to the assumption on end of infectiousness and decreased with increasing duration of infectiousness. These results showed that PCV-2 transmission is influenced by contact structure that appears worth being taken into account in an epidemic model. PMID- 18413132 TI - Biological activity of EDQM CRS for Interferon alfa-2a and Interferon alfa-2b - assessment in two in vitro bioassays. AB - The European Directorate for the Quality of Medicines (EDQM) supplies Chemical Reference Substances (CRS) for Interferon (IFN) alfa-2a (CRS I0320300) and for IFN alfa-2b (CRS I0320301) for specified physicochemical tests. However, no information is provided as to their biological activity. In contrast, the World Health Organization (WHO) provides the 2nd International Standards (IS) for IFN alfa-2a (code 95/650) and for IFN alfa-2b (code 95/566), with activity defined in International Units (IU) for calibration of biological activity of preparations of IFN. We have compared the EDQM CRSs with the WHO ISs in two bioassay systems, one measuring the anti-proliferative activity in the Daudi cell line and the other measuring a reporter gene activation in an A549 cell line. In each of these assay systems, the CRSs gave dose - response relations, which were similar to those for the WHO ISs. Estimates of relative activity for each CRS, in terms of the respective IS, showed specific biological activity for the CRSs of the same order as the nominal specific activity for the ISs. However, the estimates of relative activity were not consistent between the two assays systems, emphasizing the need for calibration within each system, if the CRS were to be used as a working standard for bioassays. For structure-activity studies, both physicochemical and biological activity characterisation are required for the same biopharmaceutical preparation. CRS I0320300 and CRS I0320301 may prove useful as working standards for some bioassay systems. PMID- 18413133 TI - Collaborative study for the establishment of a candidate equine influenza subtype 2 American-like strain A/EQ/South Africa/4/03 - horse antiserum biological reference preparation. AB - In 2004, the Office International des Epizooties (OIE) Expert Surveillance Panel on equine influenza recommended that the American lineage component (H3N8) of equine influenza vaccines (A/eq/Newmarket/1/93-like) be updated to an A/eq/South Africa/4/03-like virus. As a consequence the common European Pharmacopoeia (Ph. Eur.) - OIE reference for equine influenza subtype 2 American-like antiserum had to be complemented by an antiserum raised in horses against an A/eq/South Africa/4/03 strain. An international collaborative study run by the European Directorate for the Quality of Medicines (EDQM) in the frame of its Biological Standardisation Programme (BSP) under the aegis of the Ph. Eur. and the OIE was organised. The study was aimed at evaluating a candidate reference horse anti serum using the single radial haemolysis (SRH) and haemagglutination inhibition (HI) tests. The standard was to be established for use in immunogenicity and batch potency assay of equine influenza vaccines as a Ph. Eur. BRP and for use in clinical diagnostic tests as an OIE-approved International Standard. The evaluation performed in the collaborative study enabled the suitability of the candidate to be demonstrated and an SRH value to be assigned. The candidate was adopted as a BRP by the Ph. Eur. Commission and approved by the OIE Biological Standards Commission as an International Standard Serum in June and September 2006, respectively. PMID- 18413134 TI - Pre-validation study for testing of avian viral vaccines for extraneous agents by PCR. AB - The biological nature of vaccines imposes a permanent risk for contamination with extraneous agents. Therefore, testing of vaccines for freedom from extraneous agents is essential in the manufacturing process and quality control. Relevant methods for testing for extraneous agents of avian viral vaccines are specified in the monographs of the European Pharmacopoeia (Ph. Eur.). Currently, most of these methods involve the use of embryonated eggs or chickens. Polymerase chain reaction (PCR) is a widely used and suitable tool for the amplification and detection of extraneous nucleic acids. Different PCR assays have been developed for the application in routine testing of veterinary vaccines. However, before introduction of new methods in monographs of the Ph. Eur., they must undergo validation. Here we report about a pre-validation study performed in Official Medicines Control Laboratories (OMCLs). Diluted samples of avian infectious laryngotracheitis, avian infectious bronchitis and avian infectious bursal disease viruses have been analysed using standardised procedures and reagents. The study demonstrated that PCR methods can be transferred to other laboratories. The results also show that further work is warranted for full validation of the method. PMID- 18413135 TI - Establishment of a replacement batch for heparin sodium biological reference preparation. AB - An international collaborative study was organised to replace the current European Pharmacopoeia biological reference preparation for heparin sodium. The project was organised by the European Directorate for the Quality of Medicines & HealthCare in the frame of its Biological Standardisation Programme. A suitable candidate batch representative of the quality of heparin products currently marketed in Europe was donated to the EDQM and included in a collaborative study involving 19 laboratories from 10 European countries, the Americas, Australia and the Council of Europe. Laboratories were requested to perform their routine assays following the prescriptions of the Ph. Eur. for the assay and the identification of unfractionated heparin and for the assay of protamine. The results made it possible to demonstrate that the candidate batch was suitable for its intended use and it was therefore established by the European Pharmacopoeia Commission as the Ph. Eur. heparin sodium BRP batch 3 in June 2007. PMID- 18413136 TI - Establishment of replacement batches for heparin low-molecular-mass for calibration CRS, and the International Standard Low Molecular Weight Heparin for Calibration. AB - An international collaborative study involving fourteen laboratories has taken place, organised by the European Directorate for the Quality of Medicines & HealthCare (EDQM) with National Institute for Biological Standards & Control (NIBSC) (in its capacity as a World Health Organisation (WHO) Laboratory for Biological Standardisation) to provide supporting data for the establishment of replacement batches of Heparin Low-Molecular-Mass (LMM) for Calibration Chemical Reference Substance (CRS), and of the International Reference Reagent (IRR) Low Molecular Weight Heparin for Molecular Weight Calibration. A batch of low molecular-mass heparin was donated to the organisers and candidate preparations of freeze-dried heparin were produced at NIBSC and EDQM. The establishment study was organised in two phases: a prequalification (phase 1, performed in 3 laboratories in 2005) followed by an international collaborative study (phase 2). In phase 2, started in March 2006, molecular mass parameters were determined for seven different LMM heparin samples using the current CRS batch and two batches of candidate replacement material with a defined number average relative molecular mass (Mn) of 3,700, determined in phase 1. The values calculated using the candidates as standard were systematically different from values calculated using the current batch with its assigned number-average molecular mass (Mna) of 3,700. Using raw data supplied by participants, molecular mass parameters were recalculated using the candidates as standard with values for Mna of 3,800 and 3,900. Values for these parameters agreed more closely with those calculated using the current batch supporting the fact that the candidates, though similar to batch 1 in view of the production processes used, differ slightly in terms of molecular mass distribution. Therefore establishment of the candidates was recommended with an assigned Mna value of 3,800 that is both consistent with phase 1 results and guarantees continuity with the current CRS batch. In phase 2, participants also determined molecular weight parameters for the seven different LMM heparin samples using both the 1st IRR (90/686) and its Broad Standard Table and the candidate World Health Organization (WHO) 2nd International Standard (05/112) (2nd IS) using a Broad Standard Table established in phase 1. Mean molecular weights calculated using 2nd IS were slightly higher than with 1st IRR, and participants in the study indicated that this systematic difference precluded establishment of 2nd IS with the table supplied. A replacement Broad Standard Table has been devised on the basis of the central recalculations of raw data supplied by participants; this table gives improved agreement between values derived using the 1st IRR and the candidate 2nd IS. On the basis of this study a recommendation was made for the establishment of 2nd IS and its proposed Broad Standard Table as a replacement for the 1st International Reference Reagent Low Molecular Weight Heparin for Molecular Weight Calibration. Unlike the 1st IRR however, the candidate material 2nd IS is not suitable for use with the method of Nielsen. The candidate materials were established as heparin low-molecular-mass for calibration batches 2 and 3 by the Ph. Eur. Commission in March 2007 and as 2nd IS low-molecular-weight heparin for molecular weight calibration (05/112) by the Expert Committee on Biological Standardization in November 2007. PMID- 18413137 TI - Collaborative study for the establishment of erythropoietin BRP batch 3. AB - The European Pharmacopoeia (Ph. Eur.) monograph on Erythropoietin concentrated solution (1316) specifies that identification and assay are performed using pharmacopoeial methods requiring the use of a reference preparation. To replace the current erythropoietin Biological Reference Preparation (BRP) of Ph. Eur., in 2006, the European Directorate for the Quality of Medicines undertook a collaborative study designed to establish a replacement batch. In order to guarantee continuity, the formulation of the candidate batch was similar to that of previous batches (1 and 2). The methods chosen to qualify the new standard were those included in the current monograph. The study was defined to allow calibration of the candidate by in vivo bioassay in terms of the current World Health Organization (WHO) International Standard (IS) and to assign a unitage. The suitability of the candidate preparation to serve as a reference standard for the other pharmacopoeial analytical procedures was also investigated. The collaborative study involved 16 laboratories from Europe, Australia, Canada, China, Japan, South Korea and the United States of America. Participants carried out biological and physicochemical assays on the candidate erythropoietin BRP batch 3 (cBRP3), using batch 2 (BRP2) and where necessary the 2nd World Health Organization International Standard (WHO 2nd IS) for recombinant erythropoietin as the reference standards. It was demonstrated that the replacement batch is appropriate for use as erythropoietin BRP in the context of the control of erythropoietin concentrated solutions according to the Ph. Eur. monograph (1316). However as regards the potency of BRP2 and cBRP3 in the mouse bioassay unexpected observations were made. Direct calibration of BRP2 against the WHO 2nd IS yielded, in all laboratories, results that were systematically higher than the potency of 32,500 IU/vial assigned by direct calibration against the WHO 2nd IS in the former study. It was therefore recommended to assign the potency of cBRP3 against BRP2, using the average of all results that were not considered as outlying obtained in the collaborative study, in order to guarantee continuity of unitage between the successive BRP batches. The outcome of the study enabled the Ph. Eur. Commission to establish the proposed standard as 'erythropoeitin BRP batch 3' (BRP3). BRP3 was established in June 2007 for use as a reference preparation for the polycythaemic and normocythaemic mouse bioassay, with an assigned potency of 35,280 IU/vial, the identification by capillary zone electrophoresis, by polyacrylamide gel electrophoresis, immunoblotting and peptide mapping and as a reference for checking the system suitability of size exclusion chromatographic procedures used in the test for dimers and related substances of higher molecular mass in the Ph. Eur. monograph (1316). PMID- 18413138 TI - Fibrillogenesis in ADan peptides is inhibited by biphenyl ethers. AB - In this study, biphenyl ethers of diverse functionality were used to assess their effect on fibrillogenesis of both the oxidized and reduced ADan peptides, in vitro. It was noted that these compounds not only stalled fibrillogenesis but were also able to disrupt pre-formed fibers. The EC(50) values for the inhibition of this process lie in the nanomolar range for 50 microM of peptide concentration, indicating the high potency of these compounds as inhibitors. It was found that these compounds impart to the peptides, an alpha-helical conformation which does not allow them to aggregate and form fibrils. These studies also point out that the transition of peptides through alpha-helical conformation may be a prelude to the onset of fibrillogenesis for oxADan peptides. PMID- 18413139 TI - Enhanced anti-tumor effects achieved in a murine tumor model using combination therapy of recombinant human manganese superoxide dismutase and adriamycin. AB - Manganese superoxide dismutase (MnSOD) is the only primary antioxidant enzyme in mitochondria that scavenges superoxide radicals. Overexpressing MnSOD in cancer cells by cDNA transfection suppresses tumor formation and reverses malignant growth. In this study, we examined the effect of recombinant human manganese superoxide dismutase (rhMnSOD) alone and in combination with adriamycin (ADR) against solid tumors of sarcoma 180 in Institute of Cancer Research (ICR) mice. Administration of rhMnSOD alone and in combination with ADR significantly inhibited tumor growth in a dose-dependent manner. The use of rhMnSOD in combination with ADR enhanced ADR's anti-tumor potency without increasing toxicity. Histopathological examination provided evidence of the anti-tumor effect. In addition, we found lymphocyte infiltration of the tumors, with an increase in both CD4- and CD8-positive cells in the treated tumors. The expression of CD4 and CD8 was up-regulated with increasing dose of rhMnSOD, and the combination treatment with ADR further enhanced this up-regulation. Collectively, these data indicate that rhMnSOD may exhibit an anti-tumor effect by stimulating the immune system and promoting the recruitment of lymphocytes into the tumor to kill tumor cells. Thus MnSOD may constitute a potential new therapeutic agent to be exploited as an adjuvant in cancer therapy. PMID- 18413140 TI - alpha-Actinin links LPP, but not zyxin, to cadherin-based junctions. AB - The actin regulator VASP localizes to cell-cell junctions and has been implicated in cell-cell adhesion. VASP is recruited to sites of actin dynamics by interactions with proline rich FPPPPP motifs. Zyxin and its relative LPP use FPPPPP motifs to recruit VASP to specific cellular locations, thus directing changes in actin dynamics. It has been proposed that zyxin and LPP localize to cell-cell junctions by binding alpha-actinin. However, the role of alpha-actinin in recruiting zyxin and LPP to cell-cell contacts has not been experimentally tested. Here we use zyxin and LPP fragments to demonstrate that the alpha-actinin binding site of both proteins independently targets to cell-cell junctions. While the alpha-actinin binding site is required for LPP localization and function at cell-cell contacts, zyxin localization and function at cell-cell contacts is independent of the alpha-actinin binding site. Perturbation of LPP function, but not that of zyxin, results in changes in anchoring of alpha-actinin to detergent insoluble networks at cell-cell contacts. PMID- 18413141 TI - Calbindin-D28K prevents drug-induced dopaminergic neuronal death by inhibiting caspase and calpain activity. AB - Calbindin-D28K protects against apoptotic and necrotic cell death; these effects have been attributed to its ability to buffer calcium. In this study, we investigated the mechanisms underlying the neuroprotective effects of calbindin D28K in staurosporine (STS)-induced apoptosis and 1-methyl-4-phenylpyridinium (MPP(+))-induced necrosis. Treatment of the dopaminergic neuronal cell line MN9D with STS or MPP(+) induced cell death that was associated with increased levels of free intracellular calcium. However, only MPP(+)-induced death was inhibited by co-treatment of the cells with a calcium chelator or a sodium/calcium antiporter inhibitor. Overexpression of calbindin-D28K prevented MPP(+)-induced MN9D cell death, which occurs in the absence of any detectable caspase activation. These pro-survival effects of calbindin-D28K were associated with the inhibition of calcium-mediated calpain activation, as determined by processing of Bax. Overexpression of calbindin-D28K also blocked STS-induced MN9D death. However, this effect was accompanied by the inhibition of capase-3 cleavage, poly(ADP-ribose)polymerase cleavage, and caspase activity. These findings suggest that calbindin-D28K protects against both types of cell death by inhibiting caspase- or calcium-mediated death signaling pathway. PMID- 18413142 TI - Peptide array-based screening of human mesenchymal stem cell-adhesive peptides derived from fibronectin type III domain. AB - Human mesenchymal stem cell-adhesive peptides were screened based on the amino acid sequence of fibronectin type III domain 8-11 (FN-III(8-11)) using a peptide array synthesized by the Fmoc-chemistry. Using hexameric peptide library of FN III(8-11) scan, we identified the ALNGR (Ala-Leu-Asn-Gly-Arg) peptide that induced cell adhesion as well as RGDS (Arg-Gly-Asp-Ser) peptide. After incubation for 2h, approximately 68% of inoculated cells adhere to the ALNGR peptide disk. Adhesion inhibition assay with integrin antibodies showed that the ALNGR peptide interacts with integrin beta1 but not with alphavbeta3, indicating that the receptors for ALNGR are different from RGDS. Additionally, the ALNGR peptide expressed cell specificities for adhesion: cell adhesion was promoted for fibroblasts but not for keratinocytes or endotherial cells. The ALNGR peptide induced cell adhesion and promoted cell proliferation without changing its property. It is therefore useful for the construction of functional biomaterials. PMID- 18413143 TI - Differential phosphorylation of calreticulin affects AT1 receptor mRNA stability in VSMC. AB - The AT1 receptor plays a pivotal role for the pathogenesis of hypertension and atherosclerosis. AT1 receptor expression is regulated posttranscriptionally via destabilization of the AT1 receptor mRNA by mRNA binding proteins. Recently, we identified calreticulin as a novel binding protein within the 3'untranslated region of the AT1 receptor mRNA. Calreticulin phosphorylation is essential for binding of the AT1 receptor mRNA. In crosslink experiments, we identified src kinase as the key enzyme for calreticulin phosphorylation. Overexpression of src sense DNA resulted in vascular smooth muscle cells (VSMC) in destabilization, overexpression of src antisense resulted in stabilisation of the AT1 receptor mRNA. Furthermore, phosphorylation/dephosphorylation sites of calreticulin and their impact on the AT1 receptor mRNA stability were investigated. VSMC were stimulated with AngII before tyrosine phosphorylation as well as serine phosphorylation of calreticulin were analysed via immunoprecipitation. Stimulation of VSMC with AngII resulted in enhanced tyrosine and reduced serine phosphorylation. Both effects are essential for AT1 mRNA stability as assessed by use of pharmacological inhibitors of serine dephosphorylation (cantharidin/ocadaic acid) or tyrosine phosphorylation (tyrphostin/orthovanadat). These findings imply an important role of serine dephosphorylation and tyrosine phosphorylation on calreticulin mediated AT1 receptor mRNA stability. PMID- 18413144 TI - On the mechanisms and putative pathways involving neuroimmune interactions. AB - Bidirectional interdependence between the immune system and the CNS involves the intervention of common cofactors. Cytokines are endogenous to the brain, endocrine and immune systems. These shared ligands are used as a chemical language for communication. Such interaction suggests an immunoregulatory role for the brain, and a sensory function for the immune system. Interplay between the immune, nervous and endocrine systems is associated with effects of stress on immunity. Cytokines are thus capable of modulating responses in the CNS, while neuropeptides can exert their effects over cellular groups in the immune system. One way is controlled by the HPA axis, a coordinator of neuroimmune interactions that is essential to unravel in order to elucidate vital communications in a manner that this crosstalk remains a cornerstone in perpetuating a stance of homeostasis. PMID- 18413145 TI - The PPARgamma coding region and its role in visceral obesity. AB - Peroxisome proliferator-activated receptor gamma (PPARgamma) is a ligand activated transcription factor, plays many essential roles of biological function in higher organisms. The PPARgamma is mainly expressed in adipose tissue. It regulates the transcriptional activity of genes by binding with other transcription factor. The PPARgamma coding region has been found to be closest to that of monkey in ours and other research groups. Thus, monkey is a more suitable animal model for future PPARgamma studying, although mice and rat are frequently being used. The PPARgamma is involved in regulating alterations of adipose tissue masses result from changes in mature adipocyte size and/or number through a complex interplay process called adipogenesis. However, the role of PPARgamma in negatively regulating the process of adipogenesis remains unclear. This review may help we investigate the differential expression of key transcription factor in adipose tissue in response to visceral obesity-induced diet in vivo. The study may also provide valuable information to define a more appropriate physiological condition in adipogenesis which may help to prevent diseases cause by negative regulation of the transcription factors in adipose tissue. PMID- 18413146 TI - ABD56 causes osteoclast apoptosis by inhibiting the NFkappaB and ERK pathways. AB - We have previously shown that the biphenylcarboxylic acid butanediol ester (ABD56) inhibits osteoclast formation and activity in vitro and in vivo. However, the mechanism of action of this compound is unknown. ABD56 inhibited osteoclast formation and caused osteoclast apoptosis, but had no effects on osteoblasts or macrophages. As the NFkappaB and MAPK pathways are essential for osteoclast formation and survival, we studied the effects of ABD56 on these pathways. ABD56 caused phosphorylation of p38, JNK and nuclear translocation of c-jun in osteoclasts. ABD56-induced apoptosis was prevented by the caspase inhibitor zVAD fmk but was not prevented by the p38- or JNK-inhibitors. ABD56 completely abolished RANKL-induced IkappaB and ERK1/2 phosphorylation. Increasing the amount of RANKL partially rescued ABD56-induced apoptosis, indicating that the apoptosis is most probably due to the inhibition of survival signals such as ERK and NFkappaB, rather than activation of the p38 or Jnk MAPK pathways. PMID- 18413147 TI - Heterogeneic nature of adult cardiac side population cells. AB - Side population cells have been found in various types of adult tissue including heart and are presumed to be tissue-specific stem/progenitor cells. In the present study, we confirmed the presence of cardiac side population (cSP) cells, which showed both the Hoechst 33342 efflux ability and ABCG2 expression, in adult murine heart. Flow cytometric analysis showed that more than half of cSP cells expressed the endothelial marker VE-cadherin or the smooth muscle markers, alpha smooth muscle actin and desmin. In addition, immunohistochemical analysis demonstrated that ABCG2(+) cells were mainly localized within vascular walls. Quantitative RT-PCR analysis demonstrated that VE-cadherin(-) cSP cells progressively expressed Nkx2.5 and cardiac troponin T with time in culture. VE cadherin(-) cSP cells also expressed mesodermal-mesenchymal-associated markers and differentiated into osteocytes and adipocytes. These results highlight the heterogeneic nature of cSP cells, consisting of vascular endothelial cells, smooth muscle cells, and mesenchymal stem/progenitor cells including potential cardiomyogenic cells. PMID- 18413148 TI - Liver X receptor agonist downregulates hepatic apoM expression in vivo and in vitro. AB - It has been demonstrated that apolipoprotein M (apoM), a recently discovered HDL apolipoprotein, has antiatherosclerotic properties, which may be mediated by the enhancement of reversed cholesterol transportation and/or hepatic cholesterol catabolism. The detailed mechanisms are unknown yet. Liver X receptor (LXR) belongs to the nuclear receptor superfamily and is a ligand-activated transcription factor involved in the regulation of lipid metabolism and inflammation. Activation of LXR in the cell cultures results in an enhancement of cholesterol efflux to apoAI. In the present study, we investigated effects of the LXR agonist, T0901317 on hepatic apoM expression in vivo and in vitro. Serum apoM levels in mice given T0901317 at 10 mg or 100 mg/kg for 7 days were reduced by 12 17% (P<0.05). In HepG2 cell cultures, apoM mRNA levels were significantly lower in presence of 25 microM T0901317 (37.1%) than in control cells (P<0.001). A similar reduction was found by the addition of 9-cis retinoic acid (RA). Twenty five micromolar T0901317 together with 100 nM RA decreased apoM mRNA expression by 65% (P<0.001). Thus, the LXR agonist T0901317 significantly downregulates apoM mRNA expression in vivo and in vitro, which indicates that apoM is another novel target gene regulated by the LXR. The combination of RA and T0901317 showed additive effects, which suggests that apoM expression can be modulated by LXR/RXR pathway. PMID- 18413149 TI - Parental investment and its sensitivity to corticosterone is linked to melanin based coloration in barn owls. AB - Behavioral and physiological responses to unpredictable changes in environmental conditions are, in part, mediated by glucocorticoids (corticosterone in birds). In polymorphic species, individuals of the same sex and age display different heritable melanin-based color morphs, associated with physiological and reproductive parameters and possibly alternative strategies to cope with variation in environmental conditions. We examined whether the role of corticosterone in resolving the trade-off between self-maintenance and reproductive activities covaries with the size of melanin-based spots displayed on the ventral body side of male barn owls. Administration of corticosterone to simulate physiological stress in males revealed pronounced changes in their food provisioning rates to nestlings compared to control males. Corticosterone-treated males with small eumelanic spots reduced nestling provisioning rates as compared to controls, and also to a greater degree than did corticosterone-treated males with large spots. Large-spotted males generally exhibited lower parental provisioning and appear insensitive to exogenous corticosterone suggesting that the size of the black spots on the breast feathers predicts the ability to cope with stressful situations. The reduced provisioning rate of corticosterone treated males caused a temporary reduction in nestling growth rates but, did not affect fledgling success. This suggests that moderately elevated corticosterone levels are not inhibitory to current reproduction but rather trigger behavioral responses to maximize lifetime reproductive success. PMID- 18413150 TI - Human chorionic gonadotropin (a luteinizing hormone homologue) decreases spatial memory and increases brain amyloid-beta levels in female rats. AB - Numerous studies have suggested that estradiol (E) improves spatial memory as female rats with E perform better than those without E. However there is an inverse relationship between E and luteinizing hormone (LH) levels and LH could play a role. We examined whether treatment with the LH homologue human chorionic gonadotropin (hCG), would impair spatial memory of adult E-treated female rats. In the object location memory task, ovariectomized (ovxed) rats treated with E and either a single high dose (400 IU/kg) or a lower repeated dose of hCG (75 IU/kg hourly for 8 h) showed spatial memory disruption compared to ovxed rats treated with estradiol alone. Impairment was attributed to memory disruption as performance improved with shortened delay between task exposure and testing. Tests on another spatial memory task, the Barnes maze, confirmed that hCG (400 IU/kg) can impair memory: although E+veh treated animals made significantly fewer hole errors across time, E+hCG-treated did not. In humans, high LH levels have been correlated with Alzheimer's disease (AD). Because brain amyloid-beta (Abeta) species have been implicated as a toxic factor thought to cause memory loss in AD, we analyzed whether hCG-treated animals had increased Abeta levels. Levels of Abeta from whole brains or hippocampi were assessed by Western blot. hCG treatment to E-implanted females significantly increased soluble Abeta40 and Abeta42 levels. These results indicate that high levels of LH/hCG can impair spatial memory, and an increase in brain Abeta species may account for the memory impairment in hCG-treated rats. PMID- 18413152 TI - Association of haptoglobin phenotypes with markers of diabetic nephropathy in Type 2 diabetes mellitus. AB - BACKGROUND: Recent studies have suggested an inheritable influence of haptoglobin phenotypes on the development of microalbuminuria in diabetic patients. In this study, we aimed to investigate whether microalbuminuria and serum levels of Cystatin C (Cys C) were correlated with haptoglobin phenotypes in patients with Type 2 diabetes mellitus. METHODS: Fasting blood samples and first morning urine specimens were collected from 216 patients with type 2 diabetes (mean age 60.8+/ 6.8) and from 108 healthy controls (mean age 59.3+/-6.5 years). Serum was used for the determination of Cys C, creatinine, and haptoglobin (Hp) phenotypes, while urine specimens were used for the determination of microalbuminuria. Microalbuminuria was expressed as albumin (mg)/creatinine (g) ratio [ACR (mg/g)], and patients were divided into three groups according to the values of ACR: normoalbuminuria (<30), microalbuminuria (30-300) and macroalbuminuria (>300). RESULTS: Irrespective of Hp phenotypes, Cys C, but not creatinine, was significantly higher in patients than that in controls. According to haptoglobin phenotypes, however, the levels of Cys C, duration of diabetes, and age, were significantly higher in patients with Hp1-1 phenotype than that in patients with the other phenotypes. Moreover, micro- and/or macroalbuminuria were present in all diabetic patients, with no significant difference in frequency of occurrence among the three Hp phenotypes. Multiple linear regression revealed that duration of diabetes and age of patient were significant predictors of Cys C (P<.0001), whereas creatinine associated with age only (P=.016). Logistic regression analysis showed that duration of diabetes [P< or =.0001; odds ratio (OR)=1.678, 95% CI=1.43-1.97], and age (P=.0004; OR=0.847, 95% CI=0.77-0.93), significantly associated with micro- and/or macroalbuminuria. Hp phenotypes however, were not associated with any marker of kidney function. CONCLUSION: Findings from this study demonstrate no association between Hp phenotypes and markers of diabetic nephropathy. Markers of diabetic nephropathy however, are greatly influenced by duration of diabetes and age of patients. PMID- 18413151 TI - Changes in mood, cognitive performance and appetite in the late luteal and follicular phases of the menstrual cycle in women with and without PMDD (premenstrual dysphoric disorder). AB - Although it's been reported that women with premenstrual dysphoric disorder (PMDD) have increased negative mood, appetite (food cravings and food intake), alcohol intake and cognitive deficits premenstrually, few studies have examined these changes concurrently within the same group of women or compared to women without PMDD. Thus, to date, there is not a clear understanding of the full range of PMDD symptoms. The present study concurrently assessed mood and performance tasks in 29 normally cycling women (14 women who met DSM-IV criteria for PMDD and 15 women without PMDD). Women had a total of ten sessions: two practice sessions, 4 sessions during the follicular phase and 4 sessions during the late luteal phase of the menstrual cycle. Each session, participants completed mood and food related questionnaires, a motor coordination task, performed various cognitive tasks and ate lunch. There was a significant increase in dysphoric mood during the luteal phase in women with PMDD compared to their follicular phase and compared to Control women. Further, during the luteal phase, women with PMDD showed impaired performance on the Immediate and Delayed Word Recall Task, the Immediate and Delayed Digit Recall Task and the Digit Symbol Substitution Test compared to Control women. Women with PMDD, but not Control women, also showed increased desire for food items high in fat during the luteal phase compared to the follicular phase and correspondingly, women with PMDD consumed more calories during the luteal phase (mostly derived from fat) compared to the follicular phase. In summary, women with PMDD experience dysphoric mood, a greater desire and actual intake of certain foods and show impaired cognitive performance during the luteal phase. An altered serotonergic system in women with PMDD may be the underlying mechanism for the observed symptoms; correspondingly, treatment with specific serotonin reuptake inhibitors (SSRIs) remains the preferred treatment at this time. PMID- 18413153 TI - The effect of simvastatin on the serum monocyte chemoattractant protein-1 and intracellular adhesion molecule-1 levels in diabetic rats. AB - OBJECTIVE: This study aimed to observe the effect of simvastatin on the serum monocyte chemoattractant protein-1 (MCP-1) and intracellular adhesion molecule-1 (ICAM-1) levels and to probe its protective mechanisms on macroangiopathy in diabetic rats. METHODS: Twenty-four Wistar rats were randomly assigned to a normal control group (Group A, n=8), and STZ-induced diabetic group (Group B, n=8), or a simvastatin-treated diabetic group (Group C, n=8). Rats in Group C were treated with simvastatin (20 mg kg(-1) day(-1)) 1 week after the establishment of the diabetic model. Groups A and B were treated with corresponding sodium chloride. Peripheral blood glucose was tested weekly; serum MCP-1, ICAM-1, and HbA1c levels were tested at the eighth week. RESULTS: At the second, fourth, and eighth week, peripheral blood glucose levels in Group B were similar to those of Group C, which were much higher than those of Group A. Serum MCP-1 and ICAM-1 levels in Groups B and C were higher than those of Group A (P<.01), and serum MCP-1 and ICAM-1 levels in Group C were lower than those of Group B (P<.01); HbA1c was not significantly different between Group C and Group B. CONCLUSION: Simvastatin has the effect of anti-inflammation, which may play some protection against the progress of atherosclerosis in diabetic rats. PMID- 18413154 TI - Hand disorders, hand function, and activities of daily living in elderly men with type 2 diabetes. AB - AIMS/HYPOTHESIS: This study aimed to examine hand disorders, symptoms, overall hand function, activities of daily living (ADLs), and life satisfaction in elderly men with type 2 diabetes mellitus (DM), impaired glucose tolerance (IGT), and normal glucose tolerance (NGT). METHODS: Subjects were interviewed and evaluated with a battery of clinical and laboratory tests, including hand assessment, and a questionnaire. RESULTS: HbA1c differed between groups (highest in DM, especially in long-term DM). Limited joint motion (LJM), for example, prayer sign and Dupuytren's contracture, was most common in individuals with DM, followed by individuals with IGT, as compared to those with NGT. Vibrotactile sense was impaired symmetrically in the index and little fingers in DM. However, there were no differences for sensibility, dexterity, grip strength, and cold intolerance between groups. Individuals with long-term (>15 years) DM were more affected regarding sensibility and ADL than individuals with short-term DM, who had more sleep disturbances. ADL difficulties were less among IGT subjects. Vibrotactile sense showed correlations with Semmes-Weinstein monofilament test and static two-point discrimination. CONCLUSIONS/INTERPRETATION: Dupuytren's contracture and impaired vibrotactile sense in finger pulps occurred in patients with DM but not in those with IGT, although LJM occurred in both IGT and DM patients. A longer duration of DM was associated with more severe neuropathy and ADL difficulties. Life satisfaction was high, and hand disorders did not have a significant impact on ADL. PMID- 18413155 TI - The effects of sulfonylurea glyburide on superoxide dismutase, catalase, and glutathione peroxidase activities in the brain tissue of streptozotocin-induced diabetic rat. AB - OBJECTIVES: A member of the second-generation sulfonylureas, glyburide (GLY; glibenclamide) provides an effective therapy for patients with type 2 diabetes. It stimulates pancreatic insulin secretion, suggesting that it is effective in the treatment of type 2 diabetes primarily by elevating the circulating insulin levels. However, experimental evidences have indicated that sulfonylureas have also had an extrapancreatic effect, which may directly contribute toward maintaining blood glucose homeostasis during diabetes. METHODS: In this study, we administrated GLY to streptozotocin-induced diabetic rats and determined the effects of such treatment on activities of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) from brain tissue. RESULTS AND DISCUSSION: Brain CAT and GPx activities were not significantly different in the diabetic group compared to controls (P>.05), but the SOD activity was significantly reduced in the diabetic group compared to controls (P<.001). GLY treatment of 4 weeks had restored the SOD and CAT enzyme activities in diabetic rat brain (P<.05). In addition, high blood glucose levels of untreated diabetic rats were decreased following the GLY treatment (P<.01). Administration of GLY to diabetic rats restored the diabetes-induced changes, suggesting that GLY could restore the brain SOD and CAT activities. PMID- 18413156 TI - Candida albicans osteomyelitis in a diabetic foot ulcer. PMID- 18413157 TI - The kidney expression of matrix metalloproteinase-9 in the diabetic nephropathy of Kkay mice. AB - BACKGROUND: Disorder of the extracellular matrix has been suggested to play a role in the development of diabetic nephropathy. The aim of the present study was to examine the expression of matrix metalloproteinase-9 (MMP-9) in the kidney of Kkay mice, a type 2 diabetic model. METHODS: Eight Kkay control mice (KA) and eight Kkay diabetic mice (KB) were fed with the same diets for 4 months and sacrificed after glucose levels and weight were measured. The renal pathological changes were examined by periodic acid-Schiff and periodic acid-silver methenamine staining, and the glomerulosclerosis indexes (GIs) were calculated. The levels of MMP-9 protein in the kidney tissue were assessed by immunohistochemistry. The gene expression of MMP-9 was measured by reverse transcription polymerase chain reaction. RESULTS: The diabetic group had morphological findings of diabetic nephropathy, and their GI (240.0+/-17.3) was much higher than that of the control group (118.4+/-15.1) (P<.01). The mean positive area of MMP-9 protein in the glomeruli of Kkay mice in the KB group was higher than that in the KA group. The expression of MMP-9 messenger RNA level in the diabetic KB group was up-regulated when compared with the KA group. CONCLUSION: The expression of MMP-9 in the kidney of mice with diabetic nephropathy was enhanced, and MMP-9 might be involved in the development of diabetic nephropathy. PMID- 18413158 TI - The incidence of heart failure among nondiabetic patients with and without impaired fasting glucose. AB - OBJECTIVE: The purpose of this study was to elucidate the relationship between fasting plasma glucose (FPG), development of diabetes, and incident heart failure (HF) in a large, community sample of nondiabetic subjects. RESEARCH DESIGN AND METHODS: From Kaiser Permanente Northwest medical records, we identified 10,113 subjects with an FPG level of 100-125 mg/dl in 1997 or 1998 who were free of diabetes and HF and matched them to an equal number of subjects with an FPG level of <100 mg/dl on sex and 5-year age groups. Subjects were followed until a new diagnosis of HF was entered into the medical record, death, termination of health plan membership, or December 31, 2005, whichever came first. RESULTS: After controlling for known HF risk factors, each 10 mg/dl increase in FPG was independently associated with an 8% increase in the risk of HF over a mean follow up of 79 months [hazard ratio (HR)=1.08, 95% confidence interval (CI) 1.03-1.13, P=.003]. However, in a subsequent analysis that included only those HF cases that occurred prior to diabetes onset and censored follow-up at the time of diabetes development, FPG was not a significant predictor of HF risk (HR=1.01, 95% CI 0.96 1.07, P=.621). Age, male sex, body mass index, smoking, and cardiovascular disease were highly predictive of HF incidence. CONCLUSIONS: Although the risk of HF is increased among subjects with higher FPG, the increased risk is explained by greater likelihood of developing diabetes. Risk factors other than FPG are much stronger independent predictors of incident HF. PMID- 18413159 TI - Factors affecting hospitalization costs in Type 2 diabetic patients. AB - OBJECTIVE: To evaluate Type 2 diabetes hospitalization costs and their determinants by applying a proper methodological approach, taking into account the presence of several observations with zero costs. METHODS: A cohort study using per-patient hospital discharge abstracts in a period of 4.5 years of follow up (from 1/1/1996 to 30/6/2000). Potential cost predictors were age, sex, body max index, hypertension, diabetes duration, hemoglobin A1c levels, insulin treatment, retinopathy, coronary artery disease, peripheral artery disease, nephropathy, death and presence of comorbidity (cancer, chronic liver disease, chronic obstructive pulmonary disease, and psychiatric disease). Among risk factors, total cholesterol, HDL cholesterol and smoking were considered. A two part model has been adopted in order to take into account the presence of patients with zero hospital costs: the probability of any hospitalization has been modeled via a standard logit generalized linear model (GLM); the actual level of total costs has been modeled via a GLM, with a gamma cost distribution and a LOG link function. RESULTS: In 4.5 years the median total cost per hospitalized person was $4404 (mean $8180). In line with existing evidence, diabetes complications showed a high impact on average costs. In particular, peripheral and coronary artery diseases determined more than $1000 increase in the median costs. Chronic comorbidity were responsible for the highest incremental hospitalization costs ($1771). CONCLUSIONS: Hospitalization costs were significantly increased by the presence of diabetes complications and chronic conditions. The adoption of a two-part model has allowed to obtain estimates not neglecting the effect of covariates on the probability of having no hospital care. PMID- 18413160 TI - Effect of bitter gourd and spent turmeric on glycoconjugate metabolism in streptozotocin-induced diabetic rats. AB - AIM: Changes in glycoconjugate metabolism during the development of diabetic complications and their modulation by feeding bitter gourd and spent turmeric as fiber-rich source. METHOD: This was studied by measuring the contents of total sugar, uronic acid, amino sugar, and sulfate in the streptozotocin-induced diabetic rats. RESULTS: Total sugar content decreased in liver, spleen, and brain, while an increase was observed in heart and lungs. Uronic acid content in liver, spleen, and brain decreased, and marginal increase was observed in testis. Amino sugar content decreased in liver, spleen, lungs and heart during diabetes, and augmentation was observed to different extents. Decrease in sulfation of glycoconjugates was observed in liver, spleen, lungs and heart during diabetes and was significantly ameliorated by bitter gourd and spent turmeric, except brain. Protein content decreased in liver, while an increase was observed in brain. The studies clearly showed alteration in glycoconjugate metabolism during diabetes and amelioration to different extents by feeding bitter gourd and spent turmeric. CONCLUSION: Improvement is due to slow release of glucose by fiber in the gastrointestinal track and short-chain fatty acid production from fiber by colon microbes. PMID- 18413161 TI - Effect and safety of spinal cord stimulation for treatment of chronic pain caused by diabetic neuropathy. AB - AIM: Spinal cord stimulation (SCS) has been shown effective as a therapy for different chronic painful conditions, but the effectiveness of this treatment for pain as a result of peripheral diabetic neuropathy is not well established. The primary objectives of this study were to evaluate the effect and safety of SCS for treatment of pain and the effects on microcirculatory blood flow in the affected areas in patients with refractory peripheral diabetic neuropathy. METHOD: The study was designed as a prospective, open-label study. Data were collected during screening, at implant and at regular intervals, after initiation of therapy. Eleven diabetic patients with chronic pain in their lower limbs and no response to conventional treatment were studied. The SCS electrode was implanted in the thoracic epidural space. Neuropathic pain relief was assessed by Visual Analogue Scale (VAS) and microcirculatory skin perfusion was measured with Laser Doppler flowmetry. RESULTS: Nine subjects had significant pain relief with the percutaneous electrical stimulator. Average pain score for all nine patients was 77 at baseline and 34 at 6 months after implantation. At the end of the study, eight of nine patients continued to experience significant pain relief and have been able to significantly reduce their pain medication. For six of them, the stimulator was the sole treatment for their neuropathic pain. No significant changes in microcirculatory perfusion were recorded. CONCLUSION: Spinal cord stimulation offers an effective and safe therapy for chronic diabetic neuropathic pain. PMID- 18413162 TI - Association of the angiotensinogen M235T and angiotensin-converting enzyme insertion/deletion gene polymorphisms in Turkish type 2 diabetic patients with and without nephropathy. AB - OBJECTIVE: Recent studies have suggested an association between a deletion variant of the angiotensin-converting enzyme (ACE) gene and diabetic nephropathy. However, this finding has not been confirmed by all investigators. Furthermore, an M235T variant of the angiotensinogen (AGT) gene has been associated with hypertension, an important risk factor for the development and progression of diabetic nephropathy. RESEARCH DESIGN AND METHODS: We investigated the relationship of the ACE insertion/deletion (I/D) and AGT M235T gene polymorphisms in Turkish patients with type 2 diabetes mellitus (DM) with and without diabetic nephropathy. A total of 102 individuals were screened for the presence of the ACE I/D and AGT M235T polymorphism: 46 individuals who had type 2 DM with diabetic nephropathy and, as controls, 56 individuals who had type 2 DM without diabetic nephropathy. Gene polymorphisms were determined by the specific melting temperature (T(m)) values of the resulting amplicons after real-time online polymerase chain reaction and melting curve analysis. RESULTS: The frequencies of the ACE DD, ID, and II genotypes were 34.8%, 37.0%, and 28.3%, respectively, among type 2 diabetic patients with nephropathy, and 33.9%, 42.9%, 23.2%, respectively (P=.788), in the control subjects without diabetic nephropathy. On the other hand, the frequencies of the AGT MM, MT, and TT genotypes among the same groups were 26.1%, 52.2%, 21.7% and 26.8%, 57.1%, 16.1%, respectively (P=.758). CONCLUSIONS: There were no differences in the frequencies of the AGT M235T and ACE I/D genotypes between Turkish patients with type 2 DM with and without nephropathy. PMID- 18413163 TI - Increasing severity of cardiac autonomic neuropathy is associated with increasing prevalence of nephropathy, retinopathy, and peripheral neuropathy in Turkish type 2 diabetics. AB - BACKGROUND: Cardiac autonomic neuropathy (CAN) is the most frequent and clinically important form of diabetic autonomic neuropathy. CAN is associated with increased frequency of other microvascular complications of diabetes mellitus (DM). In this study, we evaluated the prevalence diabetic nephropathy, retinopathy, and peripheral neuropathy, and measured gastric emptying time and bladder emptying time in type 2 diabetic patients with varying degrees of CAN. PATIENTS AND METHODS: Fifty-three patients with DM complicated by CAN (30 women and 23 men; mean age, 58.8+/-9.15 years; duration of diabetes, 13.9+/-7.7 years) were included in this study. The patients were grouped according to the degree of CAN as early, definite, and severe CAN. RESULTS: There were significant differences regarding the prevalence of nephropathy, retinopathy, and peripheral neuropathy diagnosed by EMG among those groups (P<.05). However, there was no significant difference regarding the prevalence of peripheral neuropathy diagnosed by neurological examination (P>.05), and scintigraphic measurements of gastric and bladder emptying time were comparable among the groups (P>.05). CONCLUSION: The prevalence of other diabetic microvascular complications increase as the severity of CAN increases in patients with type 2 DM. This study emphasizes the need for an early screening for peripheral neuropathy, retinopathy, and nephropathy in type 2 diabetic patients with CAN, especially with severe involvement. PMID- 18413164 TI - Effects of mitemcinal (GM-611), an orally active erythromycin-derived prokinetic agent, on delayed gastric emptying and postprandial glucose in a new minipig model of diabetes. AB - AIMS: This study was conducted to evaluate the suitability of a new minipig model for investigating aspects of diabetes such as delayed gastric emptying and glucose metabolism abnormalities, and to test the effects of mitemcinal (GM-611), an orally active erythromycin-derived motilin receptor agonist, on gastric emptying and postprandial glucose in normal and diabetic minipigs. METHODS AND RESULTS: Intravenous injection of 300 mg/kg streptozotocin (STZ) to 5-week-old minipigs induced moderate hyperglycemia (about 200 mg/dl) for >80 weeks without insulin treatment. Decreased insulin production (P<.05), increased area under the glucose curve (P<.05), and slower glucose disappearance (P<.05) were demonstrated, and there was no severe inhibition of body weight gain, liver failure, or renal failure. Gastric emptying was significantly delayed in diabetic minipigs (P<.05) at 80 weeks, but not at 40 weeks, post-STZ. Oral administration of mitemcinal (5 mg/kg) at 80 weeks accelerated gastric emptying and induced a similar postprandial glucose profile in normal and diabetic minipigs with delayed gastric emptying. CONCLUSIONS: The new diabetic minipig model showed suitability for investigating diabetes, gastric emptying, and plasma glucose excursions. Since delayed gastric emptying and irregular plasma glucose excursions are characteristic of diabetic gastroparesis, the accelerating and regulating effects of mitemcinal on this model add to the existing evidence that mitemcinal is likely to be useful for treating diabetic gastroparesis. PMID- 18413165 TI - Noninvasive blood glucose monitoring using the optical signal of pulsatile microcirculation: a pilot study in subjects with diabetes. AB - OBJECTIVE: The objective of this study was to determine the conditions for optimizing measurements obtained with a noninvasive blood glucose monitor using the optical signal of pulsatile microcirculation (OSPM) in both prediabetic and diabetic subjects receiving medication. RESEARCH DESIGN AND METHODS: Eighteen subjects (3 prediabetic, 15 diabetic) aged 61.8 [15.9] years (mean [S.D.]) were studied. OSPM was the pulsatile component (P) of the signal obtained and analyzed by a blood glucose monitor. The measurement was calibrated to the fingerstick meter for each subject for personal calibration. Data were obtained from all subjects using both meters. RESULTS: A total of 179 data pairs were measured and analyzed. The validity of the position of the tested finger was assessed using the position criterion, which resulted in the removal of 38 data pairs. The criterion for the intensity of the P signal was satisfied by 141 data pairs, with nonconforming data (with a much lower P signal) mainly occurring below 26 degrees C. A total of 113 data points passed both criteria, and 100% of them fell within Zones A and B of the Clarke error grid. Data in Zones A and B exhibited a linear relationship (r=.81; slope=0.82; intercept=28.0) between noninvasive and fingerstick measurements. CONCLUSIONS: Environmental temperature has the greatest influence on the capability of the OSPM technique to monitoring blood glucose concentration, which is subject dependent. The position of the tested finger is the second major factor, hence a carefully designed finger adaptor is essential. PMID- 18413166 TI - Evaluation of in vivo cerebral metabolism on proton magnetic resonance spectroscopy in patients with impaired glucose tolerance and type 2 diabetes mellitus. AB - The aim of this study was to investigate possible metabolic alterations in cerebral tissues on magnetic resonance spectroscopy (MRS) in patients with impaired glucose tolerance (IGT) and with type 2 diabetes mellitus (T2-DM). Twenty-five patients with T2-DM, 13 patients with IGT, and 14 healthy volunteers were included. Single-voxel spectroscopy (TR: 2000 ms, TE: 31 ms) was performed in all subjects. Voxels were placed in the frontal cortex, thalamus, and parietal white matter. N-acetylaspartate (NAA)/creatine (Cr), choline (Cho)/Cr, and myo inositol (MI)/Cr ratios were calculated. Frontal cortical Cho/Cr ratios were increased in patients with IGT compared to control subjects. Parietal white matter Cho/Cr ratios were significantly higher in patients with IGT when compared to patients with T2-DM. In the diabetic group, frontal cortical MI/Cr ratios were increased, and parietal white matter Cho/Cr ratios were decreased when compared to the control group. Frontal cortical NAA/Cr and Cho/Cr ratios and parietal white matter Cho/Cr ratios were decreased in diabetic patients with poor glycemic control (A1C>10%). A1C levels were inversely correlated with frontal cortical NAA/Cr and Cho/Cr ratios and with parietal white matter Cho/Cr ratios. T2-DM and IGT may cause subtle cerebral metabolic changes, and these changes may be shown with MRS. Increased Cho/Cr ratios may suggest dynamic change in membrane turnover in patients with IGT. Diabetic patients with poor glycemic control may be associated with neuronal dysfunction/damage in brain in accordance with A1C levels and, in some, extend with insulin resistance. PMID- 18413167 TI - Elevated third-trimester haemoglobin A 1c predicts preterm delivery in type 1 diabetes. AB - The prevalence of preterm delivery is considerably elevated in women with type 1 diabetes. The aim of the study was to evaluate haemoglobin A(1c) (HbA(1c)) as a predictor of preterm delivery. Two hundred thirteen consecutive pregnant women with type 1 diabetes and normal urinary albumin excretion were included prospectively. HbA(1c) was analyzed at 10, 20 and 28 weeks of gestation. Seventy one women (33%) delivered pre term and 142 at term. At 10 weeks of gestation, HbA(1c) was 7.3% (S.D. 1.0) vs. 6.9% (S.D. 0.9) (P<.01), at 20 weeks of gestation 6.6% (S.D. 0.7) vs. 6.1% (S.D. 0.7) (P<.001) and at 28 weeks of gestation 6.7% (S.D. 0.8) vs. 6.1% (S.D. 0.7) (P<.001). When comparing HbA(1c) at 10, 20 and 28 weeks of gestation, HbA(1c) at 28 weeks of gestation (P<.001) was the best predictor of preterm delivery. The adjusted odds ratio per 1% increment in HbA(1c) at 28 weeks of gestation was 2.8 (95% CI 1.7-4.4). HbA(1c) at 28 weeks of gestation was a clinical significant predictor of preterm delivery in type 1 diabetes. PMID- 18413168 TI - Psychosocial factors contributing to persistent depressive symptoms in type 2 diabetic patients: a Croatian survey from the European Depression in Diabetes Research Consortium. AB - OBJECTIVE: The study was aimed at exploring a 1-year course of depression in persons with type 2 diabetes and analysing demographic, disease-related, and psychological variables that may predict persistent depressive symptoms. PATIENTS AND METHODS: One hundred patients from a randomly selected sample of 470 outpatients were found to be suffering from severe depressive symptoms. They were followed and re-examined for depression after 1 year. Baseline depression was assessed by the Center for Epidemiologic Studies--Depression scale (CES-D) and a face-to-face diagnostic interview relying on the DSM-IV. Nonparametric tests for between-group differences were used to compare patients who recovered from depression with those who still suffered from severe depressive symptoms. Multiple logistic regression was used to determine predictors of depression persistence. RESULTS: Seventy-nine of 100 patients with baseline depression scores indicative of severe depression were reached at 1-year follow-up. Among them, 53% were shown to have improved depressive symptoms to CES-D <16, while 47% continued to suffer from severe depressive disturbances (CES-D >or=16). Logistic regression analysis indicated that psychosocial variables predicted persistently elevated depressive symptoms better than demographic and diabetes-related ones. Clinical depression at baseline (OR=3.8, CI 1.31-10.98, P=.01), diabetes-related distress (OR=3.3, CI 1.01-10.98, P=.05), and social and physical quality-of-life aspects (OR=0.92, CI 0.88-0.97, P=.0005 and OR=0.94, CI 0.90-0.98, P=.002, respectively) were shown to be independent predictors of 1-year depression outcomes. CONCLUSIONS: Severity of baseline depression, a degree to which depression disrupted the patients' quality of life, and concomitant emotional problems related to diabetes were shown to be associated with persistently elevated depressive symptoms. PMID- 18413169 TI - Pheochromocytoma presenting as diabetic ketoacidosis. AB - Pheochromocytoma is a rare form of secondary hypertension and may be potentially lethal if left untreated. The classical symptoms are paroxysmal hypertension, headaches, palpitations and sweating. They are caused by sudden catecholamine release. Hyperglycemia is reported in some patients with pheochromocytoma but diabetic ketoacidosis is an extremely rare complication of pheochromocytoma with only four cases reported. We report a case of a young woman with pheochromocytoma manifested as diabetic ketoacidosis. PMID- 18413170 TI - Synergistic interaction between semicarbazide-sensitive amine oxidase and angiotensin-converting enzyme in diabetes: functional analysis by gene ontology. AB - Plasma semicarbazide-sensitive amine oxidase (SSAO) and angiotensin-converting enzyme (ACE) were studied for their correlation with diabetes (DM) complication. The effect of interaction between SSAO and ACE in DM complication is of interest. Studying the functional change due to interaction between SSAO and ACE is difficult. In this work, the author used a new gene ontology technology to predict the functional change resulting from the interaction between SSAO and ACE. According to this study, there is a synergetic effect resulting from the interaction between SSAP and ACE. This can imply that co-expression of SSAP-ACE leads to more severe complication of DM. However, the author can also demonstrate that some molecular functions such as proteasome activator activity of SSAO and hydrolase activity, metallopeptidase activity, and zinc ion binding of ACE are suppressed after co-expression. These results provide good information for further study in diabetes medicine. However, further experimental works are required to support this in silico research. PMID- 18413171 TI - Postural blood pressure changes and associated factors in long-term Type 1 diabetes: Wisconsin Epidemiologic Study of Diabetic Retinopathy. AB - PURPOSE: To describe the frequency of orthostatic hypotension and hypertension and associations with risk factors in a cohort of persons with long-term Type 1 diabetes (n=440) participating in the Wisconsin Epidemiologic Study of Diabetic Retinopathy. METHODS: Evaluations included detailed medical history, electrocardiography (ECG), and laboratory tests. Blood pressure (BP) was measured in supine and standing positions. Standing decrease in systolic (SBP) or diastolic (DBP) BP of at least 20 or 10 mmHg, respectively, was defined as orthostatic hypotension; increase of SBP from <140 to >or=140 mmHg or DBP from <90 to >or=90 mmHg was defined as orthostatic hypertension. RESULTS: Prevalence of orthostatic hypotension and orthostatic hypertension was 16.1% and 15.2%, respectively. Some ECG measurements of cardiac autonomic dysfunction were significantly associated with orthostatic hypotension. Association between SBP and orthostatic hypotension and orthostatic hypertension were significant [odds ratio, 1.02 (95% confidence interval, or CI, 1.01-1.05) and 1.02 (95% CI, 1.01 1.04), per 1 mmHg, respectively] after adjusting for confounders. Interaction between SBP and age was observed. SBP was significantly associated with orthostatic hypotension and orthostatic hypertension in people 25 kg/m(2)), with poor glycemic control (hemoglobin A(1c) >7%), but free from overt diabetic vascular complications. Anthropometric parameters, blood pressure (BP), cardiorespiratory fitness, glycemic and lipid profile, fibrinogen, plasminogen activator inhibitor-1, high-sensitivity C-reactive protein (hsCRP), insulin resistance, and 24-h urinary albumin excretion (UAE) were measured. Based on the median Vo(2) peak value, participants were placed into low fitness (LF, n=46) or moderate fitness group (MF, n=46). RESULTS: In univariate analysis, exercise capacity correlated with systolic (r=-0.349) and diastolic BP (r=-0.441), waist circumference (r=-0.345), total cholesterol (r= 0.348), high-density lipoprotein (HDL) (r=0.362), UAE (r=-0.486), homeostasis model assessment (HOMA-IR) (r=-0.467), uric acid (r=-0.316), and hsCRP (r=-0.217, only in women subgroup) (P<.05). With the exception of the latter three variables, the above associations remained significant after controlling for age and sex in multiple regression analysis (P<.05). Compared to LF group, patients in MF group showed significantly higher levels of HDL and lower levels of BP, waist circumference, hsCRP, and HOMA-IR (P<.05). In addition to this, UAE tended to be lower in fit patients (P=.054). CONCLUSIONS: Low cardiorespiratory fitness seems to be independently associated with most traditional and emerging cardiovascular risk factors in patients with T2DM. Even a moderate increase of cardiorespiratory fitness exerts beneficial effects on cardiovascular risk profile. PMID- 18413174 TI - The effect of hemodialysis on accelerated atherosclerosis in diabetic patients: correlation of carotid artery intima-media thickness with oxidative stress. AB - OBJECTIVE: Both diabetes and hemodialysis (HD) are associated with increased oxidative stress. The aim of this study was to clarify the effect of maintenance HD on oxidative stress parameters in diabetic patients and to explore any relation between carotid artery intima-media thickness (CIMT) and oxidative stress markers. METHODS: Twenty Type 2 diabetic patients undergoing chronic maintenance HD, 20 type 2 diabetic patients with normal renal function, and 20 age- and sex-matched healthy subjects were included. Serum thiobarbituric acid reactive substances (TBARS), protein carbonyl content (PCO), and nitrite/nitrate levels were determined as oxidative stress markers. Serum vitamin E, plasma sulfhydryl (P-SH), erythrocyte glutathione (GSH) levels, and superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) activities were measured as antioxidants. CIMT was assessed by carotid artery ultrasonography. RESULTS: Both diabetic patient groups had enhanced oxidative stress indicated by higher levels of TBARS, PCO, and nitrate/nitrite and lower activities of SOD, CAT, and GPx compared to controls. Diabetic patients undergoing HD had significantly higher CIMT (P=.001) and higher levels of nitrite/nitrate (P=.05), PCO (P=.03), and GSH (P=.04) but significantly lower levels of P-SH (P<.001), serum vitamin E (P=.04), SOD (P=.02), CAT (P=.001), and GPx (P=.006) compared to diabetic patients with normal renal functions. There were significant negative correlations between CIMT and SOD (r=-0.50, P<.001), CAT (r=-0.41, P=.003), and P SH levels (r=-0.51, P<.001) and significant positive correlation between CIMT and nitrite/nitrate levels (r=0.41, P=.003) and TBARS (r=0.35, P=.02). Linear regression analysis showed TBARS was significantly and positively correlated with CIMT (P=.04), while SOD and P-SH were significantly and negatively correlated with CIMT (P=.05 and P=.02, respectively). CONCLUSIONS: Hemodialysis exacerbates oxidative stress and disturbances in antioxidant enzymes in diabetic patients. Serum nitrite/nitrate and TBARS can be used as positive determinants, while erythrocyte SOD, CAT activities, and P-SH level can be used as negative determinants of atherosclerosis assessed by CIMT in diabetic patients. PMID- 18413175 TI - The effects of two different hypocaloric diets on glucagon-like peptide 1 in obese adults, relation with insulin response after weight loss. AB - OBJECTIVE: Few studies have investigated the effect of type of diets on GLP-1 concentrations. The aim of this study was to compare the effect of two diets on circulating GLP-1 levels and the relation with insulin response after weight loss. METHODS: A population of 118 obese patients were analyzed. Patients were randomly allocated to two groups: (a) Diet I (low carbohydrate) and (b) Diet II (low fat). Biochemical and anthropometric parameters were measured before and after 3 months of hypocaloric diet. RESULTS: Fifty-two patients (12 male/40 female) were treated with Diet I and 66 patients (21 male/45 female) with Diet II. In Group I, basal GLP-1 levels did not change after dietary treatment (9.4+/ 3.3 vs. 9.9+/-3.1 ng/ml; ns). In Group II, GLP-1 levels decreased significantly (8.4%) (9.2+/-3.3 vs 8.7+/-3.1 ng/ml; P<.05). In the multivariate analysis with a dependent variable (levels of GLP-1), only insulin levels remained as an independent predictor in the model (F=5.9; P<.05), with an increase of 0.6 ng/ml (95% CI 0.1-1.1) GLP-1 concentrations with each increase of 1 mUI/ml of insulin. CONCLUSION: A hypocaloric diet with a low fat percentage decreased GLP-1 levels with a direct correlation with insulin levels. Nevertheless, patients with a hypocaloric diet with a low carbohydrate percentage treatment did not change GLP 1 levels. Diet macronutrient manipulation on GLP-1 response could be useful in an obesity nutrition therapy. PMID- 18413176 TI - Mortality in diabetic and nondiabetic patients after amputations performed from 1996 to 2005 in a tertiary hospital population: a 3-year follow-up study. AB - AIMS: Diabetes is the leading cause of lower-extremity amputations worldwide. The objective of this study was to look at the survival after first amputation between subjects with and without diabetes in a sample of Greek population. METHOD: We performed a retrospective study of all nontrauma, nonneoplasm-related amputations performed in a tertiary centre during the years 1996-2005 in diabetic (n=183) and nondiabetic patients (n=75). Survival status was assessed from the first amputation until December 31, 2005. RESULTS: A total of 54.6% of amputees with diabetes and 51.6% of those without diabetes died in a mean [95% confidence interval (CI)] time of 4.3 (3.5-5.1) and 6.6 (4.6-8.6) years after the first amputation, respectively (P=.65). Diabetic patients underwent a second amputation (P=.003) and contralateral amputations (P=.02) more often in comparison with nondiabetic subjects. Predictors of all-cause mortality in the diabetic group, after adjustment for sex, were age [hazard ratio (HR) (95% CI), 1.04 (1.02-1.06); P<.001] and the level of amputation (major vs. minor) [HR, 1.55 (1.00-2.40), P=.05]. The respective values in the nondiabetic patients were HR of 1.06 (1.03 1.08; P<.001) and HR of 3.12 (1.27-7.64; P=.01). Median length of hospital stay was comparable between the two groups. CONCLUSION: Mortality rates after amputation were high in both patients with and without diabetes. Older age and a higher level of amputation were associated with poorer survival. Diabetic patients more often underwent a second amputation to the same and the contralateral limb. Additionally, mortality rates, length of hospital stay, and perioperative mortality were not different between patients with and without diabetes. PMID- 18413177 TI - Endothelial dysfunction and history of recurrent depression in postmenopausal women with Type 2 diabetes: a case-control study. AB - OBJECTIVES: This study of postmenopausal women with Type 2 diabetes mellitus (T2DM) investigated (1) history of depression as a predictor of endothelium dependent flow-mediated dilation (FMD); (2) the relative associations of single and recurrent depressive disorders with FMD; and (3) cortisol as a potential mechanism. METHODS: Participants were nonsmoking, naturally postmenopausal women with T2DM with no known vascular disease. All were free of current mood disorder. On average, the 44 participants were 63 years of age, White, diabetic for 6 years, and were in adequate glycemic control. Thirty-eight percent were never depressed, 19% had experienced one disorder, and 43% had experienced recurrent disorders. History of depression was assessed with Structured Clinical Interview for Diagnostic and Statistical Manual-IV. Current depressive symptoms were measured with Center for Epidemiological Studies Depression (CESD) scale. FMD was assessed by standard procedures and calculated as percent change in brachial artery diameter from baseline. RESULTS: Women with history of recurrent depression showed vasoconstriction (mean=-1%), which was significantly different from women with history of single depression (mean=+6) and never depressed women (mean=+5) (P<.05), both of whom showed similar levels of vasodilation. In logistic regression controlling for hypertension, duration of diabetes, and glycemic control, history of recurrent depressive disorders predicted greater likelihood of vasoconstriction (P<.05, odds ratio=4.23) but history of single depressive disorder did not. Controlling for current depressive symptoms did not account for effects of past recurrent depressive disorders. Cortisol was not related to FMD. CONCLUSIONS: In postmenopausal women with T2DM, recurrent depressive disorders, even in full remission, are associated with endothelial dysfunction. Potential mechanisms of the relationship between depression and endothelial dysfunction other than cortisol warrant investigation. PMID- 18413178 TI - Iron status in women with and without gestational diabetes mellitus. AB - OBJECTIVE: Gestational diabetes mellitus (GDM) affects approximately 7% of all pregnancies. Pregnancy, mostly because of the mitochondria-rich placenta, is a condition that favors oxidative stress. A transitional metal, especially iron, which is particularly abundant in the placenta, is important in the production of free radicals. Also, studies have shown that free radicals have a role in GDM. As there are little data about iron status in GDM, this study was performed to compare iron status in GDM and control group. RESEARCH DESIGN AND METHODS: In this case-control study, 34 women with diagnosed GDM were compared with 34 non GDM women in the control group at 24-28 weeks of pregnancy in terms of iron status, including ferritin, serum iron, total iron-binding capacity (TIBC), hemoglobin (Hb), mean corpuscular volume (MCV), and mean corpuscular hemoglobin (MCH). RESULTS: In this study, concentration of serum ferritin, iron, transferrin saturation and hemoglobin, MCV, and MCH was significantly higher in the GDM group and TIBC was significantly lower in this group (P<.05). No significant association was observed with the other variables including familial history of diabetes and GDM. CONCLUSION: Our findings indicate an association between increased iron status and GDM. The role of iron excess from iron supplementation in the pathogenesis of GDM needs to be examined. PMID- 18413179 TI - Racial differences in diabetes self-management and quality of care in Texas. AB - AIM: To assess racial/ethnic differences in diabetes self-management behaviors and quality of care in Texas. METHODS: This cross-sectional study assessed self management behaviors and quality of care in 1720 adults with diabetes in the 2002 2004 Texas Behavioral Risk Factor Surveillance Survey. Multiple logistic regression models were used for assessing the independent association between race/ethnicity, self-management behaviors, and quality of care variables controlling for covariates. SAS (SAS Institute Inc, 2002-2003) was used for statistical analysis. RESULTS: Eighteen percent of Hispanics, 14% of Blacks, and 10% of Whites reported never performing home glucose testing. Seventeen percent of Hispanics, 11% of Blacks, and 10% of Whites reported never doing home foot exam. Thirty-two percent of Hispanics, 21% of Blacks, and 16% of Whites did not have an A1C test in the prior 12 months. Twelve percent of Hispanics, 10% of Blacks, and 6% of Whites did not have a dilated eye exam in the prior 12 months. Fifty-four percent of Whites, 42% of Blacks, and 40% of Hispanics received a flu shot. Forty-nine percent of Whites, 30% of Blacks, and 26% of Hispanics received a pneumonia shot. In adjusted models, Hispanics were more likely to be sedentary (OR, 1.64; 95% CI, 1.08-2.49) compared to Whites. Blacks did not differ significantly from Whites (OR, 1.46; 95% CI, 0.92-2.34). Hispanics (OR, 1.61; 95% CI, 1.01-2.57) and Blacks (OR, 1.83; 95% CI, 1.10-3.03) were more likely to get an annual foot exam by a provider compared to Whites. Hispanics (OR, 0.54; 95% CI, 0.34-0.85) and Blacks (OR, 0.58; 95% CI, 0.35-0.98) were less likely to get a pneumonia shot compared to Whites. CONCLUSIONS: Hispanics have poorer access to care and poorer health status compared to Whites or Blacks. Controlling for socioeconomic and access to care variables eliminated disparities in self management but did not eliminate disparities in quality of diabetes care. PMID- 18413180 TI - Clinical resolution of glycogenic hepatopathy following improved glycemic control. AB - Glycogenic hepatopathy is an underrecognized complication of long-standing poorly controlled diabetes mellitus. It is characterized by abnormal glycogen accumulation in hepatocytes, elevated liver enzymes and hepatomegaly. This is a distinct entity from other causes of hepatomegaly and elevated liver enzymes in diabetics such as nonalcoholic fatty liver disease. It is important to distinguish this entity as it has the potential for resolution following improved glycemic control as described by this case report. PMID- 18413181 TI - Ethnic and gender differences in psychosocial factors, glycemic control, and quality of life among adult type 2 diabetic patients. AB - OBJECTIVE: To examine ethnic and gender differences in psychosocial factors, e.g., social support and acceptance of the disease, knowledge levels, perceived difficulty in adherence behaviors, and diabetes outcome (glycemic control and quality of life) in Type 2 diabetic patients. METHODS: Data were collected via telephone interviews from 180 diabetic subjects (34% Hispanics, 27% Non-Hispanic whites, 18% African-Americans, and 20% Asian-Indians; 52% females) from 2 clinics. Hemoglobin A1c levels were obtained from patient charts. RESULTS: Significant ethnic and gender differences existed in acceptance of the disease, in receiving social support, disease knowledge, perceived difficulty in self management behaviors, glycemic control, and quality of life among Type 2 diabetic patients; differences were more pronounced by gender than by ethnicity. In general, social support and acceptance of the disease were high. However, perceived difficulty in self-management behaviors varied by racial/ethnic groups with self-monitoring of blood glucose perceived as most difficult by Hispanic respondents, dietary management was most difficult for non-Hispanic whites, while physical activity was the most difficult for African-Americans. Hispanic respondents had greatest severity of the disease (poorest metabolic control). CONCLUSIONS: Ethnic and gender variations exist in social support, acceptance of diabetes, quality of life, and adherence behaviors. The outcomes of diabetes care can be improved if practitioners factor these differences in tailoring diabetes education and supportive care for individuals with Type 2 diabetes. PMID- 18413182 TI - Hepatic glycogenosis: a rare cause of hepatomegaly in Type 1 diabetes mellitus. AB - Hepatomegaly, with or without abnormal liver function tests, was a common feature of both pediatric and adult patients with diabetes mellitus. We are reporting a case of a 16-year-old diabetic boy in whom we found hepatomegaly, mildly elevated transaminases and elevated serum lipids never noticed before. Abdominal ultrasound confirmed hepatomegaly; liver biopsy pointed out a picture compatible with glycogenosis. The patient's abnormal liver function tests, elevated serum lipids and hepatomegaly decreased over a period of 4 weeks with tight metabolic control. This situation was due to overinsulinization because the patient assumed an excessive quantity of food and therefore took an excessive quantity of insulin. In conclusion, hepatomegaly may be seen in diabetic patients due to hepatic glycogen accumulation as a result of excessive food and insulin consumption. In hepatic glycogenosis, the pathological findings improve in 4 weeks when good metabolic control is provided. Therefore, the other reasons must be investigated when hepatomegaly persists for a longer period. PMID- 18413183 TI - National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK) meeting summary: Advances toward measuring diabetic retinopathy and neuropathy: from the bench to the clinic and back again (April 4-5, 2007, Baltimore, Maryland). AB - The National Institute of Diabetes and Digestive and Kidney Diseases sponsored a meeting recently to explore new ways to assess diabetic retinopathy and neuropathy, both in diabetic patients and in diabetic mice. The workshop compared current gold standards for assessment of retinopathy and neuropathy, new improvements of existing techniques, and new functional biomarkers measured with nontraditional technologies. Since the anatomical changes that comprise diabetic retinopathy and neuropathy take long to develop and have proven difficult to arrest once initiated, some talks highlighted the value of methods that are based on the pathophysiology that precedes, and might contribute to, the histopathology. In addition, a goal of the workshop was to produce a set of working criteria on phenotyping diabetic retinopathy and neuropathy that could be reviewed by the scientific community. PMID- 18413184 TI - Renal connective tissue growth factor correlates with glomerular basement membrane thickness and prospective albuminuria in a non-human primate model of diabetes: possible predictive marker for incipient diabetic nephropathy. AB - Diabetic renal disease is characterized by accumulation of extracellular matrix, glomerulosclerosis, and tubulointerstitial fibrosis. Connective tissue growth factor (CTGF) is implicated in these changes, as it contributes to new matrix synthesis and is increased in the diabetic kidney. CTGF also inhibits mesangial matrix degradation through up-regulation of the tissue inhibitor of matrix metalloproteinase 1 (TIMP-1). In a non-human primate model of diabetes, we determined whether the level of renal CTGF protein before development of albuminuria correlated with renal matrix and TIMP-1 changes and whether renal CTGF predicts progression to albuminuria. METHODS: In a group of diabetic (n=9) and control (n=6) baboons after a 5-year duration of diabetes, renal tissue CTGF and TIMP-1 were detected by immunohistochemistry and compared with glomerular basement membrane (GBM) thickness and mesangial volume measurements from electron photomicrographs of renal biopsies. Urinary albumin levels were measured at 5 and 10 years of diabetes. RESULTS: GBM thickness, CTGF protein, and TIMP-1 protein were increased after 5 years of diabetes (each P<.05). Tubular fibronectin scores correlated with tubular CTGF scores (r=0.72, P=.002). In diabetic animals, GBM thickness correlated with tubular and total CTGF levels (P=.002 and P=.04, respectively), whereas mesangial cell and total matrix volume correlated with glomerular TIMP-1 (P=.02 and P=.01, respectively). Tubular CTGF scores (P=.008) and GBM thickness (P=.03) at 5 years in diabetes each predicted the degree of albuminuria at 10 years. CONCLUSIONS: These results suggest that early increases in renal CTGF protein contribute to incipient diabetic nephropathy and that renal CTGF may have utility as an early marker for progression to dysfunction in the diabetic kidney. PMID- 18413185 TI - Peroxynitrite-induced protein nitration contributes to liver mitochondrial damage in diabetic rats. AB - Oxidative stress, especially peroxynitrite (ONOO(-))-mediated oxidative stress, plays a key role in diabetes. Mitochondria, as the generating source of ONOO(-), may also be the major damaging target of ONOO(-), which can cause a series of mitochondrial proteins nitration. Therefore, this study aimed to clarify the relationship between the nitration of entire mitochondrial proteins induced by ONOO(-) and liver mitochondrial structural damage in diabetes. Sprague-Dawley male rats were injected with streptozotocin to induce diabetes. After 10 weeks, transmission electron microscopy was used to observe the ultrastructure of liver mitochondria, and reverse transcription-polymerase chain reaction was used to detect liver inducible nitric oxide synthase (iNOS) mRNA expression. Nitrotyrosine (NT) content and distribution were detected with Western blot analysis and immunohistochemistry. In addition, some biochemical indicators were detected to represent oxidative stress and metabolic disorders. In diabetic rats, increasing levels of iNOS mRNA and NT content (P<.05) were observed, in accord with pathological alterations of the ultrastructure of liver mitochondria. Meanwhile, some alterations in biochemical indicators were observed in diabetes. Treatment with aminoguanidine could significantly attenuate these alterations (P<.01 or P<.05). In conclusion, the nitration of mitochondrial proteins induced by ONOO(-) may be responsible for structural damage to liver mitochondria, and aminoguanidine can reduce ONOO(-) generation and attenuate mitochondrial damage. PMID- 18413186 TI - Association of the -250G/A promoter polymorphism of the hepatic lipase gene with the risk of peripheral arterial disease in type 2 diabetic patients. AB - AIM: This study aimed to investigate the association between a polymorphism in the hepatic lipase (LIPC) gene promoter and the presence of peripheral arterial disease (PAD) in persons with type 2 diabetes. PATIENT AND METHODS: We evaluated 120 type 2 diabetics and identified those with PAD according to the ankle-arm index. The G-250A polymorphisms in the promoter of the LIPC gene were studied by PCR restriction. A logistic regression analysis was performed to determine the association between the rare allele and PAD. RESULTS: The prevalence of PAD was 19%. The frequency of the -250A allele was 0.211 in the group without PAD and 0.395 in the group with PAD (P<.05). Carriers of the -250A allele differed only in the ankle-arm index (0.92+/-0.12 for carriers vs. 1.00+/-0.12 for noncarriers, P<.05), with the difference remaining significant after adjustment for covariates (age; sex; waist-to-hip ratio; body mass index; duration of diabetes; smoking; hypertension; glycated hemoglobin; triglycerides; HDL cholesterol; LDL cholesterol; small, dense LDL cholesterol). Only smoking [odds ratio (OR)=6.93, 95% confidence interval (CI)=2.12-22.69, P=.001] and the -250A allele (OR=2.89, 95% CI=1.07-7.84, P=.036) were significantly associated with vascular disease in the logistic regression analysis. CONCLUSIONS: Patients with type 2 diabetes who are carriers of the rare -250A allele in the promoter of the hepatic lipase gene are susceptible to PAD. PMID- 18413187 TI - Distribution of glyoxalase I polymorphism among Zuni Indians: the Zuni Kidney Project. AB - Zuni Indians are experiencing simultaneous epidemics of type 2 diabetes mellitus (T2DM) and renal disease [Scavini, M., Stidley, C. A., Shah, V. O., Narva, A. S., Tentori, F., Kessler, D. S., et al. (2003). Prevalence of diabetes is higher among female than male Zuni Indians: Diabetes among Zuni Indians. Diabetes Care, 26 (1), 55-60; Shah, V. O., Scavini, M., Stidley, C., Tentori, F., Welty, T., Maccluer, J. W., et al. (2003). Epidemic of diabetic and nondiabetic renal disease among the Zuni Indians: The Zuni Kidney Project. Journal of the American Society of Nephrology, 14, 1320-1329]. Methylglyoxal (MG), a highly reactive, cytotoxic, cross-linking endogenous aldehyde involved in the modification of biologic macromolecules, is elevated among patients with T2DM. Glyoxalase I (Glo1) is the initial enzyme involved in the detoxification of MG. Glo1 is a dimeric enzyme with three isoforms Glo1-1, Glo2-1, and Glo2-2, resulting from a point mutation (A-->C) at position 332 of cDNA. The present study was conducted to explore the hypothesis that specific polymorphisms of the Glo1 gene are associated with diabetes and/or albuminuria in Zuni Indians. We studied four groups of Zuni Indians stratified by diabetes status and albuminuria, as assessed by the urinary albumin:creatinine ratio (UACR): Group I--normal controls; Group II--T2DM and UACR<0.03; Group III--T2DM and UACR>or=0.03; and Group IV- nondiabetic participants with UACR>or=0.03. Genomic DNA was used as template for polymerase chain reaction amplification of the Glo1 gene. Products were digested to yield 110-bp bands (homozygous, CC); 54- and 45-bp bands (homozygous, AA); or all three bands (heterozygous CA). Data on age, gender, UACR, serum creatinine, hemoglobin A1(c), serum glucose, systolic and diastolic blood pressures, and the duration of T2DM among participants in Groups II and III were analyzed using analysis of variance. A generalized linear model logistic regression analysis was used to assess the relationships between specific Glo1 polymorphisms to T2DM and UACR. All three Glo1 genotypes were present among Zuni Indians. There were no significant differences in the distributions of Glo1 genotypes among the study groups (chi-square test, P=.5590). The prevalence of Glo1 A allele was higher among diabetic participants (Groups II and III combined) than among nondiabetic participants (Groups I and IV combined) (chi-square test, P=.0233). There was an association (odds ratio=2.9; 95% confidence interval=1.3-7.2) between the Glo1 A allele and T2DM. PMID- 18413188 TI - A longitudinal study of patients with diabetes and foot ulcers and their health related quality of life: wound healing and quality-of-life changes. AB - OBJECTIVE: The objectives of this study were to assess ulcer healing in patients with diabetic foot ulcers during a 1-year follow-up, to compare health-related quality of life (HRQL) between those with healing ulcers and those with ulceration, and to assess whether changes in healing affect patients' HRQL. METHODS: Data were collected at baseline, after 6 months (T(2)), and after 12 months (T(3)). Adult patients (n=127) with type 1 or type 2 diabetes and foot ulcers were recruited from outpatient clinics. Self-assessed HRQL was measured using the SF-36 Health Survey at all measurement points. Sociodemographic and clinical data were obtained at baseline, and data on healing, ulceration, and amputation were obtained at T(2) and T(3). RESULTS: Of 127 patients included at baseline, 35% had persistent ulcer, 37% had healed ulcer, 6% had undergone amputation, and 7% had died at T(3). In addition, 9% had undergone amputation at T(2). Significant differences were observed between patients with healed ulcers and patients with persistent ulcers on general health at baseline; on physical functioning, general health, and social functioning at T(2); and on five of eight subscales in the SF-36 (e.g., physical functioning, role limitation--physical, general health, social functioning, and mental health) and on the physical summary score at T(3). Furthermore, HRQL improved significantly in social functioning and mental health in parallel with healing of ulcers, and deteriorated in the social functioning subscale in parallel with nonhealing of ulcers. CONCLUSIONS: Only 37% of ulcers healed during the 1-year follow-up. Poor HRQL during follow-up was associated with poor ulcer prognosis. PMID- 18413189 TI - The effect of polymorphisms in the renin-angiotensin-aldosterone system on diabetic nephropathy risk. AB - OBJECTIVES: The risk of diabetic nephropathy (DN) can be increased by elevated intraglomerular pressure and glomerular filtration rate, leading to glomerular damage. This can be controlled by the renin-angiotensin-aldosterone (RAA) system, which has an important function regulating both systemic and intrarenal blood pressure. Smoking increases the risk of DN, but not all diabetic patients who smoke develop DN. There is a possibility that smoking has different effects depending on the different genotypes of the individual. We investigated the association of DN with seven polymorphisms in the RAA system and their possible interaction with smoking. SUBJECTS AND METHODS: In the present case-control study, type 1 diabetic patients with diabetes duration > or =20 years, without albuminuria and without antihypertensive treatment (n=197), were included as controls. An albumin excretion rate (AER) of 20-200 microg/min (n=73) was considered as incipient DN, and an AER >200 microg/min was considered as overt DN (n=48). Smoking habits were obtained from questionnaires. RESULTS: Homozygosity for the A allele, of the angiotensin II type 1 receptor (AGTR1) A1166C polymorphism, was associated with increased risk of overt DN (OR=3.04; 99% CI=1.02-9.06), independently of the other associated variables: age, duration of diabetes, ever smoking, HbA1c, and sex. The effect of the AA genotype was enhanced to a four times risk increase among ever-smoking patients. Two alleles of the microsatellite marker adjacent to the angiotensinogen gene were less common among nephropathy cases than among controls, but this was not significant when controlling for the same variables as above. CONCLUSIONS: The risk of having overt DN was increased in patients homozygous for the A1166 allele, and smoking seemed to enhance the effect of the AGTR1 genotype. PMID- 18413190 TI - Prevalence of osteoporosis and factors affecting bone mineral density among postmenopausal Turkish women with type 2 diabetes. AB - OBJECTIVES: Controversy remains as to the effects of type 2 diabetes on bone metabolism. The aims of this study were to assess the association between type 2 diabetes and bone mineral density (BMD) and to evaluate the possible relationship between chronic diabetic complications and bone density. METHODS: Bone mineral densities at the lumbar spine, femur, and radius in 206 postmenopausal Turkish women with type 2 diabetes were evaluated by dual-energy X-ray absorptiometry and compared with those in 61 age-matched postmenopausal nondiabetic women. Medical and lifestyle characteristics, body mass index (BMI), hemoglobin A1c level, and status of microvascular and macrovascular diabetic complications were recorded. Frequency of osteoporosis and that of osteopenia as well as the relationship between microvascular and macrovascular complications and BMD were evaluated. RESULTS: The groups did not differ on BMDs and T scores at the hip, lumbar spine, and radius. Patients with radial and/or lumbar and/or hip osteoporosis had a longer duration of diabetes (P=.000), were older (P=.000), and had a lower BMI (P=.000). No correlation was found between osteopenia or osteoporosis and hemoglobin A1c level, presence of microalbuminuria, retinopathy, neuropathy, peripheral artery disease, cerebrovascular event, and coronary artery disease. Among the three sites, BMD at the hip was positively correlated with BMI (P=.000) but negatively correlated with age (P=.000) and duration of diabetes (P=.000). Presence of microalbuminuria revealed a negative correlation with BMD at the femoral neck (P=.042). CONCLUSION: There is no evidence that type 2 diabetes influenced BMD in our postmenopausal patient group. PMID- 18413191 TI - Inhibition of platelet GPIIb-IIIa and P-selectin expression by aspirin is impaired by stress hyperglycemia. AB - Increased aspirin resistance may contribute to the increase in thrombotic events observed in patients with type 2 diabetes. In this study, we examined if acute exposure to increased plasma glucose impaired the inhibitory effects of aspirin on platelet activation. Whole-blood samples were incubated with 100 (euglycemia), 200, 300, and 600 mg/dl glucose followed by incubation with aspirin [acetylsalicylic acid (ASA)]. Using flow cytometry, GPIIb-IIIa and P-selectin were analyzed in unstimulated and arachidonic acid (AA)-stimulated platelets. In euglycemic blood, AA caused a significant increase in platelet GPIIb-IIIa expression [unstimulated: 59.5+/-8.2 total fluorescence intensity (TFI), AA stimulated: 319.6+/-42.7 TFI, P=.002] and P-selectin (4.4+/-0.7 and 179.5+/-38.5 TFI, P<.001). In vitro, ASA significantly inhibited both GPIIb-IIIa expression (36.5%) and P-selectin expression (81%; P<.005). However, increased blood glucose (200 mg/dl) significantly impaired the inhibitory effect of ASA (84% for GPIIb IIIa, P<.005; 48% for P-selectin, P=NS). Increasing glucose to 600 mg/dl completely overwhelmed the inhibitory effect of ASA. A statistically significant interaction between glucose concentration and ASA dose was found (P<.001 for GPIIb-IIIa and P=.004 for P-selectin). In vitro, concentration-dependent stress hyperglycemia significantly impaired the inhibitory effects of aspirin on human platelet GPIIb-IIIa and P-selectin expression. Under acute hyperglycemic conditions, the effectiveness of ASA to inhibit platelets via the AA-activation pathway may be significantly reduced. PMID- 18413192 TI - Long-term benefits of insulin therapy and glycemic control in overweight and obese adults with type 2 diabetes. AB - PROBLEM: Obesity and type 2 diabetes have reached epidemic proportions in the United States. Obese patients are at especially high risk for the development of metabolic syndrome, a clustering of metabolic abnormalities associated with insulin resistance that usually precede the development of cardiovascular disease. Overweight or obesity, along with insulin resistance, is frequently present in people with type 2 diabetes. METHODS: A literature search of the PubMed and MEDLINE databases, using the terms diabetes, obesity, metabolic syndrome, glycemic control, antidiabetic therapy, and insulin, was performed. Articles published between 1985 and 2006 that examined diabetes management in the obese population were selected and reviewed. RESULTS: There is new evidence suggesting that tight glycemic control and earlier initiation of insulin therapy can improve outcomes in obese patients with type 2 diabetes, thereby reducing the risk for the development of both macrovascular and microvascular complications of the disease. Insulin also appears to exhibit anti-inflammatory effects, which may provide additional protection against the development of atherosclerosis. Despite the benefits of insulin therapy, many patients and physicians remain reluctant to start insulin due to concerns about weight gain. CONCLUSION: Newer insulin formulations can effectively improve glycemic control without significant effects on patient weight and, therefore, may be particularly useful in patients who are overweight or obese. Implementation of comprehensive treatment regimens that emphasize dietary modification, physical activity, and exercise, and aggressive use of pharmacological agents to achieve tight glycemic control through physiological regimens offer the most promise for reducing long-term complications in obese patients with type 2 diabetes. PMID- 18413193 TI - Hyperglycemia predicts mortality after CABG: postoperative hyperglycemia predicts dramatic increases in mortality after coronary artery bypass graft surgery. AB - OBJECTIVE: Risk of morbidity and mortality after coronary artery bypass graft surgery (CABG) is higher in patients with clinical diabetes mellitus (DM). We evaluated whether outcomes are affected by postoperative hyperglycemia in CABG patients independent of preoperative DM diagnosis. RESEARCH DESIGN AND METHODS: A total of 2297 consecutive CABG patients were studied. The first glucose value after surgery completion (mean 15 min) was tested as a predictor of outcome. Primary outcome variables were prolonged ventilation (>24 h), deep sternal wound infection, renal failure, permanent stroke, any reoperation, length of stay (>14 days) and mortality. All outcomes except for prolonged ventilation and length of stay were tracked out to 30 days postoperatively. Patients were stratified by glycemic control: Low (glucose <80), normal (referent, glucose 80-110), high (glucose 111-200) and very high (glucose >200 mg/dl). Multivariable logistic regression was used to determine the independent predictive value of glycemic groups, adjusted for outcome specific risk scores from the Society of Thoracic Surgeons model. RESULTS: Patient distribution among groups low through very high were 44 (1.9%), 476 (20.7%), 1425 (62.0%) and 352 (15.3%). Greater complication rates were noted in the very high group when compared with the referent group: prolonged ventilation (adjusted odds ratio (OR)=2.66, P<.001), length of stay >14 days (adjusted OR=2.06, P=.004) and mortality (adjusted OR=7.71, P<.001). CONCLUSION: Patients with blood glucose values >200 mg/dl immediately after CABG had an increased risk of complications, including mortality, independent of a clinical diagnosis of DM. This study documents the high risk associated with early postoperative hyperglycemia in this group, suggesting the need for prospective trials of glycemic control. PMID- 18413194 TI - Advanced glycation index and its association with severity of diabetic retinopathy in type 2 diabetic subjects. AB - BACKGROUND: This study investigates the association of advanced glycation index (AGI), a simple assay to detect advanced glycation endproducts (AGEs) in serum, with severity of diabetic retinopathy (DR) in type 2 diabetic subjects. METHODS: The study included 188 type 2 diabetic subjects without DR, 153 subjects with nonproliferative DR, 41 subjects with proliferative DR, and 188 control participants. Serum levels of AGEs were monitored with a spectrofluorimeter by recording Maillard-specific fluorescence. RESULTS: AGI values increased with severity of DR (analysis of variance, P<.0001). Among diabetic subjects, AGI (mean+/-S.E.) was higher among subjects with nonproliferative diabetic retinopathy (NPDR; 6.7+/-0.1 U) and proliferative diabetic retinopathy (PDR; 9.1+/-0.3 U) than among subjects without DR (P<.0001). By arranging the levels of serum AGI in quartiles, the proportion of PDR subjects increased with increasing AGI values, with maximum subjects in the last quartile (trend chi(2)=60.239, P<.0001). AGI was associated with NPDR even after adjusting for age, gender, duration of diabetes, and glycated hemoglobin [odds ratio (OR)=1.33; 95% confidence interval (95% CI)=1.12-1.57; P=.001]. Similarly, AGI showed a significant association with PDR even after adjusting for various risk factors (OR=2.47; 95% CI=1.75-3.47; P<.0001). Receiver-operating-characteristics curve analysis revealed that the threshold level of 8.07 U had a 78% sensitivity, an 83.6% specificity, and an 86.1% accuracy for detecting PDR. CONCLUSION: AGI showed a significant association with the severity of DR and, hence, could be used as a prognostic tool to predict the development and progression of DR. PMID- 18413195 TI - Beta-blockers have a beneficial effect upon endothelial function and microalbuminuria in African-American subjects with diabetes and hypertension. AB - BACKGROUND: Type 2 diabetes mellitus (T2DM) with microalbuminuria (MA) is associated with increased risk of cardiovascular events (CVE) that may be attenuated by angiotensin-converting enzyme inhibitors (ACEIs), unless microalbuminuria persists (PMA). African-Americans (AA) have a higher prevalence of nephropathy with suboptimal response to ACEIs. We studied the effects of beta blockers addition and comparative effects of carvedilol with metoprolol on 24-h urinary-albumin excretion (UAE) and endothelial function (EF) in AA with PMA. METHODS: Thirty-four AA 30-70 years of age with T2DM and PMA despite ACEI therapy were randomized to receive carvedilol or metoprolol in addition to ACEI and any other concurrent therapy. Carvedilol/metoprolol dose was titrated to achieve blood pressure (BP) <130/80 mm Hg. UAE and brachial-artery reactivity were studied at baseline and 12 weeks. We analyzed the effects of addition of beta blockers and whether there was any difference in response between the two beta blockers. RESULTS: Thirty-three subjects completed the study; BP decreased to <135/80 mm Hg. After 12 weeks, beta-blocker treatment resulted in significant increase in flow-mediated dilatation (FMD) from 3.5+/-1% to 8.5+/-1% (P=.004) and significant reduction in mean log-transformed UAE from 2.655 g/g Cr+/-0.087 to 2.533 g/g Cr+/-0.093 (P=.028). FMD increased by 240% (P=.033) with carvedilol and by 110% (P=.096; NS) with metoprolol. UAE decreased with carvedilol by 0.35 g/g Cr (P=.023) and with metoprolol by 0.23 g/g Cr (P=.298; NS). CONCLUSION: Our results clearly indicate that addition of beta-blockers to ACEI improves EF and reduces UAE in high-risk AA T2DM patients with PMA. Carvedilol but not metoprolol improves EF and reduces UAE in AA with identical BP control. Larger trials are needed to further elucidate the differential effects of carvedilol/metoprolol on EF and UAE and its impact on CVE in such patients. PMID- 18413196 TI - The role of inhaled insulin in the treatment of type 2 diabetes. AB - With the increasing prevalence of type 2 diabetes, it is imperative that health care providers adopt a more aggressive treatment approach to optimize glycemic control and to reduce the incidence of and costs associated with diabetes-related morbidity and mortality. Despite the well-recognized benefits of improved glycemic control in patients with type 2 diabetes, the majority of patients still have poor glycemic control. Diabetes is a progressive disease, and many patients who initially achieve glycemic control with oral therapy will eventually require the addition of insulin to maintain glycemic control. Subcutaneous injections, however, represent a barrier to patients' initiation of insulin therapy, and alternative methods of insulin delivery are likely to be viewed positively by patients and physicians. The recent approval of Exubera (Pfizer, New York, NY), a novel inhaled insulin delivery system for patients with type 1 or type 2 diabetes, may alleviate some of the barriers to the initiation of insulin. In studies in patients with type 2 diabetes, Exubera provided effective glycemic control that was comparable to that seen with subcutaneous insulin and superior to that seen with oral therapy, with a favorable safety and tolerability profile. Exubera can reduce the need for daily insulin injections and may become a useful alternative to existing treatments for the management of type 2 diabetes. Greater acceptance of new insulin delivery systems may lead to earlier introduction of insulin therapy, which should help more patients with diabetes achieve glycemic control sooner after diagnosis. PMID- 18413197 TI - Insulin resistance following cardiothoracic surgery in patients with and without a preoperative diagnosis of type 2 diabetes during treatment with intravenous insulin therapy for postoperative hyperglycemia. AB - OBJECTIVE: To assess insulin resistance postoperatively in patients with (DM) and without (nonDM) a prior diagnosis of diabetes. RESEARCH DESIGN AND METHODS: Following cardiac surgery, 122 nonDM and 33 DM were treated with insulin infusions to obtain glucose levels <110 mg dl(-1). Glucose levels, insulin infusion rates, and insulin infusion rate/glucose ratios were calculated to assess insulin resistance. RESULTS: The average blood glucose at insulin drip initiation (209 vs. 173 mg dl(-1); P<.001) and during the first 12 h (146 vs. 135 mg dl(-1); P<.05) was higher in DM, but during Hours 12-24 glucose levels were not different. The peak (5.7 vs. 4.1 U h(-1); P<.001) and average insulin drip rates (3.7 vs. 2.9 U h(-1); P<.01) were higher in DM. Insulin resistance (insulin drip rate/glucose ratio) was higher in DM during Hours 1-12 (0.029 vs. 0.022 U h( 1) mg(-1) dl(-1); P<.001), but not during Hours 12-24 (P=.57). To eliminate glucotoxicity as a cause of the insulin resistance, 23 DM patients were pair matched with 23 nonDM patients based first on glucose levels at drip initiation then by body mass index (BMI) and then catecholamine use to maintain blood pressure. The average blood glucose levels, insulin drip rates, and insulin resistance ratios were not significantly different between the pair-matched groups at all time points. CONCLUSIONS: When matched for initial glucose levels, insulin resistance is not different between DM and nonDM following cardiac surgery, likely due to the overwhelming stress response. Insulin drip protocols used postoperatively do not have to be modified for those with a prior diagnosis of diabetes. PMID- 18413198 TI - Influence of vitamin E supplementation on endothelial complications in type 2 diabetes mellitus patients who underwent coronary artery bypass graft. AB - BACKGROUND: Diabetes mellitus (DM) is associated with increased risk for complications following coronary artery bypass graft (CABG) surgery, in which tissue damage involves leukocyte-endothelial interactions mediated by endothelin 1 (ET-1) and adhesion molecules (AMs). AIM: This study compared lipids and their peroxidation product, malondialdehyde (MDA), high-sensitivity C-reactive protein (hsCRP), ET-1, platelet-selectin (P-selectin), intercellular AM-1 (ICAM-1), and vascular cell AM-1 (VCAM-1) between healthy controls and type 2 DM subjects who did not receive CABG surgery as well as those who did. Vitamin E as an adjunctive therapy in subjects who underwent CABG was evaluated. METHODS: ELISA was used to measure hsCRP, ET-1, and AMs. For all subjects, glycosylated hemoglobin (HbA(1c)) and lipid profile were estimated. RESULTS: Percentage of HbA(1c), lipids, MDA, hsCRP, ET-1, P-selectin, ICAM-1, and VCAM-1 levels were significantly higher in the diabetic groups than in healthy controls. Vitamin E supplementation for 3 successive months significantly lowered MDA, hsCRP, ET-1, ICAM-1, and VCAM-1 levels by 64%, 47%, 12%, 74%, and 25%, respectively. However, high-density lipoprotein cholesterol (HDL-C) and vitamin E serum levels were increased by 65% and 90.55%, respectively (P1.8 mmol/l) and creatinine (>115 mmol/l), smoking, proteinuria, microalbuminuria, lack of physical exercise, and retinopathy, while anti-platelet and lipid lowering drugs had lower hypertension hazard ratios. Cox proportional hazard showed that older age, male gender, higher BMI, diabetes duration (<5 years), and retinopathy were independent predictors of hypertension, while exercise, lipid lowering, and anti-platelet medications were associated with reduced hypertension incidence rate. CONCLUSIONS: Incidence of hypertension in Saudi T2DM patients is comparable to other communities, with older age, male gender, higher BMI, diabetes duration of <5 years and retinopathy being strong predictors for hypertension development. PMID- 18413200 TI - Oxidative stress pathway genes and chronic renal insufficiency in Asian Indians with Type 2 diabetes. AB - BACKGROUND: There are significant regional variations in prevalence of diabetes and diabetic chronic renal insufficiency (CRI) in India. Oxidative stress plays an important role in the development of diabetic complications. To determine the importance of the polymorphisms in the genes involved in maintenance of cellular redox balance, we performed a case control study in subjects from south and north India. METHODS: Successive cases presenting to the study centers with Type 2 diabetes of >2 years duration and moderate CRI (n=194, south India 104, north India 90) diagnosed by serum creatinine >or=2 mg/dl after exclusion of nondiabetic causes of CRI were compared with diabetes subjects with no evidence of renal disease (n=224, south India 149, north India 75). Twenty-six polymorphisms from 13 genes from the oxidative stress pathway were analyzed using polymerase chain reaction-restriction fragment length polymorphism. Genes included were superoxide dismutases (SOD1, 2, 3), uncoupling proteins (UCP1, 2), endothelial nitric oxide synthase (NOS3), glutathione-S-transferases (GST) (M1, T1, P1), vascular endothelial growth factor (VEGF), paraoxonase (PON) 1 and 2, and nicotinamide adenine dinucleotide phosphate reduced, oxidase p22(phox). Genes were tested for their association with CRI using chi(2) test. RESULTS: In south Indian (SI) subjects there was significant allelic and genotypic association of the wild-type allele in SOD2 (Ala9Val; P=.002 and P=.013, respectively), UCP1 ( 112 T>G, P=.012 and P=.009; Ala64Thr, P=.015 and P=.004), NOS3 (Glu298Asp, P=.002 and P=.009) and GSTP1 (Ile105Val, P=.003 and P=.004) genes with development of CRI. None of these observations were replicated in the north Indian (NI) subjects. A genotypic but not allelic association was observed for two markers, VEGF (-460 T>C) and PON1 (Arg192Gly) among NI diabetic CRI subjects. CONCLUSION: The nonreplication of association suggests differential genetic susceptibility of the two populations to diabetic chronic renal insufficiency. In the SI diabetic subjects, oxidative stress pathway genes might be an important predictor for the development of diabetic complications. Further, the association of wild-type alleles may suggest that they confer greater survival ability to comorbid complications and may be nephroprotective. PMID- 18413201 TI - Diabetic myocardial disease: pathophysiology, early diagnosis and therapeutic options. AB - Diabetes mellitus is a powerful risk factor for cardiovascular disease associated with high morbidity and mortality rates. Diabetic patients also have an increased incidence of heart failure which has been traditionally attributed to the concurrent presence of ischemic or hypertensive heart disease. Yet, nowadays, according to recent scientific evidence, diabetic myocardial disease (DMD) is more and more being considered as a distinct nosologic entity, independent of the co-existence of coronary artery disease, arterial hypertension or other risk factors, with the potential to lead to a self-existent progressive development of heart failure. In this article, we review the possible pathophysiologic mechanisms involved in the development of DMD as well as the structural and functional changes in the diabetic heart. We emphasize the importance of early detection of the syndrome, especially by novel echocardiographic techniques. Finally, we refer to the various therapeutic options for the optimal management of DMD according to the recent literature. PMID- 18413202 TI - Serum lipid profile in diabetic macular edema. AB - PURPOSE: To evaluate the correlation of lipid profile and clinical presentation of macular edema in Type 2 diabetes mellitus (DM) patients. MATERIALS AND METHODS: The study included 20 patients with chronic diabetic macular edema and plaque-like hard exudates (Group 1), 20 patients with diabetic macular edema (Group 2), and 20 DM patients but without retinopathy (Group 3). Diabetic retinopathy was classified according to the Early Treatment Diabetic Retinopathy Study grading system. Sample t test was used to evaluate the association between the fasting serum lipid [total cholesterol, triglyceride, low-density lipoprotein (LDL), high-density lipoprotein (HDL)], glycosylated hemoglobin (HbA1c), fasting blood glucose, creatinine levels, and the clinical findings. P values <.05 were considered statistically significant. RESULTS: There was no difference between fasting serum lipids and HbA1c levels. Duration of diabetes was shorter in Group 3 than in Groups 1 and 2. Patients in Group 1 had longer duration of diabetes than others (P<.05). Creatinine levels in Group 1 were higher than in other groups (P<.05). Although there was no correlation between fasting blood glucose and HbA1c levels, HbA1c was higher in all three groups from the baseline-normal limits (P<.05). CONCLUSION: No correlation was found between serum lipid levels and macular edema severity, but the duration of diabetes was demonstrated as a significant factor in the progression of macular edema. High HbA1c levels in all patients highlight the importance of intense glycemic control in diabetic patients. PMID- 18413203 TI - The association between erectile dysfunction and cardiovascular risk in men with Type 2 diabetes in primary care: it is a matter of age. AB - AIMS: Erectile dysfunction (ED) prevalence is usually based on questionnaires, too elaborate for daily practice. The single question for ED prevalence is unknown. Literature reports an independent association between ED and both cardiovascular disease (CVD) and diabetes. Whether routinely asking men with Type 2 diabetes (DM2) about ED identifies those at elevated risk for CVD is unknown. We assessed cardiovascular risk of DM2 men with ED. DESIGN AND METHODS: This was a cross-sectional study in primary care. During annual check-up, the practice nurse asked 1823 DM2 men: "Do you have erection problems? Yes/no." ED prevalence rate was calculated. Age, medication, and other known factors associated with ED and/or CVD were used in univariate analysis (odds ratio [OR], Student's t test, and Mann-Whitney test). This revealed confounding variables used in the multivariable analysis. The association between ED and history of cardiovascular disease (HCVD) was assessed by logistic regression analysis. In patients with no HCVD, we assessed the association between ED and 10-year United Kingdom Prospective Diabetes Study (UKPDS) coronary heart disease risk by linear regression analysis. RESULTS: The prevalence of ED in DM2 patients was 41.3%. There was no independent association between ED and HCVD [adjusted OR, 1.2 (95% CI, 0.9-1.5)]. The 10-year UKPDS CHD risk difference between men with and without ED was 5.9% (95% CI, 3.2-8.7), but after adjustment for age, this association disappeared [adjusted risk difference, 0.6% (95% CI, -1.5 to 2.7)]. CONCLUSION: The ED prevalence rate assessed by a single question was comparable to that assessed by questionnaires. ED neither did independently relate to patients' cardiovascular history nor to cardiovascular risk. PMID- 18413204 TI - Reductions in excess mortality rates among people with diabetes by selected cause of death. AB - We applied Bayesian methods to estimate excess mortality rates by selected causes of death for decedents with diabetes compared to those without diabetes in North Dakota and assessed changes in the excess rate between 1992-1998 and 1999-2003. We report the probability (Pr) of a rate decrease in the age-adjusted excess rate and considered the evidence strong if the probability was 0.90 or higher. Among men with diabetes, the evidence was strong for a probable decrease in excess rate for heart disease (8.7 per 1000 to 6.5), cerebrovascular disease (1.2 per 1000 to 0.75) and arterial disease (0.24 per 1000 to 0.08). Among women with diabetes, the evidence was strong for a probable decrease in excess rates for the overall (total) rate (17.8 per 1000 to 12.6), for heart disease (6.1 per 1000 to 4.4), IHD (4.4 per 1000 to 3.1), cerebrovascular disease (1.4 per 1000 to 0.5), arterial disease (0.17 per 1000 to 0.10) and cancer (2.1 per 1000 to 1.3) as underlying cause of death. The data reflect a high likelihood that cause-specific excess mortality is decreasing for men, and especially for women, with diabetes. PMID- 18413205 TI - Increased urinary levels of CXCL5, CXCL8 and CXCL9 in patients with Type 2 diabetic nephropathy. AB - CXC chemokines are particularly significant for leukocyte infiltration in inflammatory diseases. Recent reports have shown that inflammation is one of potential pathogenic mechanisms for diabetic nephropathy. However, information on inflammation related with CXC chemokines in human Type 2 diabetic nephropathy still remains scarce. We measured urinary and serum levels of three CXC chemokines, CXCL5, CXCL8 and CXCL9, in 45 Type 2 diabetic patients (DM), 42 primary renal disease (PRD) patients and 22 healthy controls by enzyme-linked immunosorbent assay. Urinary levels of CXCL5, CXCL8 and CXCL9 in DM were significantly elevated compared to those in controls (P<.0001, P<.01, P<.001; respectively). They increased consistent with urinary albumin excretion rate (UAER) and correlated with UAER in partial correlation analyses (r=0.41, P<.01; r=0.40, P<.01; r=0.45, P<.01; respectively). Urinary levels of CXCL5 in DM were significantly interrelated to HbA(1c) (r=0.42, P<.01). On the other hand, PRD showed significant increased levels of urinary CXCL8 and CXCL9 compared to controls (P<.001, P<.01; respectively), and so did PRD as UAER increased. However, there were no significant elevations of urinary levels of CXCL5 in PRD in spite of the increased UAER. We found significant associations of UAER in DM with diabetes duration, 1/serum creatinine, urinary CXCL5 (adjusted R(2)=0.67, P<.0001) or CXCL9 (adjusted R(2)=0.69, P<.0001) in a stepwise multiple regression analysis. These results suggest that these three CXC chemokines may be involved in the progression of human Type 2 diabetic nephropathy and that CXCL5 may be of use for telling diabetic nephropathy from primary renal diseases. PMID- 18413206 TI - Rosiglitazone protects diabetic rats against kidney disease through the suppression of renal moncyte chemoattractant protein-1 expression. AB - Although the pathogenetic mechanisms of diabetic nephropathy (DN) have not been elucidated thoroughly, an inflammatory mechanism has been suggested to contribute to its development and progression. Moncyte chemoattractant protein (MCP)-1 is a chemokine that can attract macrophages and T cells from the circulation to the local kidney, then activate them, and ultimately injure the renal tissue. Recent studies have demonstrated that thiazolidinediones decrease urinary albumin (ALB) excretion, which may be partly related to its anti-inflammatory action. Therefore, the effects of rosiglitazone on renal inflammation and renal injury were investigated in streptozotocin (STZ)-induced diabetic rats in this study. We examined the urinary excretion rates of ALB, retinal-binding protein (RBP), and MCP-1 of normal control group (Group C, n=8), STZ-induced diabetes mellitus group (Group D, n=8), and diabetes plus rosiglitazone (5 mg x kg-1 x day-1) treatment group (Group R, n=8) at the eighth week. The renal tissues of diabetic rats were obtained for reverse transcriptase-polymerase chain reaction to examine the expression of MCP-1 mRNA. Our results showed that compared to normal control, urinary excretion rates of ALB, RBP, and MCP-1 were significantly increased in untreated diabetic rats at the eighth week. However, rosiglitazone treatment could markedly decrease all the parameters above. In addition, urinary excretion rate of MCP-1 showed positive correlations with urinary ALB excretion, urinary RBP excretion, and kidney/body weight. The expressions of MCP-1 mRNA in renal tissues were markedly up-regulated in untreated diabetic rats, and these could be notably reduced by rosiglitazone treatment. In conclusion, rosiglitazone may have a potential therapeutic target in DN, which may be partly attributed to lowering of the expression of MCP-1 in the local kidney and the urinary excretion of MCP 1. PMID- 18413207 TI - Association of endothelial nitric oxide synthase Glu298Asp, 4b/a, and -786T>C gene variants with diabetic nephropathy. AB - BACKGROUND: Nitric oxide (NO) produced by endothelial NO synthase (eNOS) mediates a wide range of processes, and abnormal NO production mediated diabetes complications, including diabetic nephropathy (DN). In view of their impact on eNOS activity, polymorphisms in eNOS gene were described as candidates for atherosclerosis and DN. AIMS: We evaluated the association of -786T>C (promoter region), Glu298Asp (Exon 7), and 4b4a (Intron 4) polymorphisms in eNOS gene with Type 2 diabetes mellitus (T2DM) and DN by haplotype analysis. SUBJECTS AND METHODS: Study subjects comprised 515 DN patients, 402 normoalbuminuric [diabetes with no nephropathy (DWN)] T2DM patients, and 748 healthy subjects. -786T>C and Glu298Asp genotyping were done by PCR-RFLP analysis. RESULTS: Higher prevalence of mutant Asp298, 4a, and -786C alleles and homozygous Asp298/Asp298 and 4a/4a genotypes were seen in T2DM patients compared to healthy subjects, with increased Asp298/Asp298 seen in DN compared to DWN patients (P<.05). Three-loci haplotype analysis demonstrated significant association between eNOS variants and T2DM, with protective, neutral, T2DM, and DN-susceptible haplotypes identified, the latter including Asp298/4b/-786T and the Asp298/4a/-786C haplotypes that were present at higher frequencies among DN than among DWN patients. Multivariate regression analysis identified only Asp298/4a/-786T haplotype to be associated with DN (P=.047) after controlling for potential covariates. CONCLUSION: Genetic variation at the eNOS locus is associated with T2DM. It can serve as a useful genetic marker of increased susceptibility to T2DM and its complications, including the risk of nephropathy. PMID- 18413208 TI - The effects of Yucca schidigera and Quillaja saponaria on DNA damage, protein oxidation, lipid peroxidation, and some biochemical parameters in streptozotocin induced diabetic rats. AB - The aim of this study was to examine the effects of Yucca schidigera, Quillaja saponaria, and a mixture of both plants on streptozotocin-induced diabetic rats. Animals were allocated into five groups with 10 rats each. The control (C) and diabetic control group (D) were fed with standard rat feed (SRF). The other diabetic groups, the Y. schidigera group (DY), the Q. saponaria group (DQ), and the mix group (DQY), were fed ad libitum using SRF+100 ppm Y. schidigera powder (Sarsaponin 30), SRF+100 ppm Q. saponaria powder (Nutrafito), and SRF+100 ppm Y. schidigera-Q. saponaria powder (Nutrafito Plus), respectively, for 3 weeks. The blood glucose level was found to be significantly lower in the DY and DQ groups than in the D and DQY groups (P<.001). The insulin levels increased in the DY and DQY groups (P<.05). Plasma cholesterol and triglyceride levels in the DY, DQ, and DQY groups significantly decreased compared to those of the D group (P<.01, P<.001, respectively). HDL in the diabetic groups significantly increased in the DQ and DQY groups (P<.05), while LDL did not show any significant change. Mononuclear leukocyte DNA damage, plasma malondialdehyde, and plasma protein carbonyl levels were found to be significantly lower (P<.001, P<.001, P<.05, respectively) in the DY, DQ, and DQY groups according to the D group. The low level of nitric oxide in diabetic rats increased in the DQ group (P<.01). Total antioxidant capacity between groups did not differ. Our results thus suggested that Q. saponaria and Y. schidigera powders could help in the treatment of the disease owing to their hypoglycemic, hypocholesterolemic, and antioxidant effects. PMID- 18413209 TI - Fast and low-cost analysis of heart rate variability reveals vegetative alterations in noncomplicated diabetic patients. AB - The aim of the present preliminary study was to describe a simple protocol for the analysis of the heart rate variability (HRV) that can reveal the different autonomic control between noncomplicated diabetic patients and normal subjects within 15 min. The power spectrum of the HRV was evaluated on 5-min-long electrocardiographic recordings in both the supine and the seated positions in 30 noncomplicated non-insulin-dependent diabetic (NIDDM) patients and in 30 healthy volunteers. In healthy subjects the low-frequency (LF) value was higher in seated position than in supine position, while in diabetic patients the LF value in seated position did not differ from that in supine position and did not differ from that in healthy subjects in supine position. The present work demonstrates that the protocol described reveals a different autonomic regulation of the heart rate in healthy subjects and in NIDDM patients even if there is no clinically evident autonomic neuropathy. PMID- 18413210 TI - An increase in prevalence of diabetes mellitus in Jordan over 10 years. AB - OBJECTIVE: This study aimed to determine the prevalence of type 2 diabetes mellitus (DM) and impaired fasting glycemia (IFG), identify their associated factors, determine how the prevalence of type 2 DM has changed over 10 years, and assess the awareness and state of control of diabetes in Jordan. METHODS: Data were analyzed from a cross-sectional study that included a random sample of 1121 Jordanians aged 25 years and above. A subject was deemed affected by DM if this diagnosis was known to the patient or if his or her condition complies with the American Diabetes Association definition. IFG was defined as a fasting serum glucose level of >or=6.1 mmol/l (110 mg/dl) but <7 mmol/l. HbA(1c) >7.5% was defined as "unsatisfactory" metabolic control. RESULTS: The age-standardized prevalence of diabetes and IFG was 17.1% and 7.8%, respectively, with no significant differences between women and men. Of the 195 diabetic subjects, 146 (74.9%) had been previously diagnosed. More than half (54%) of those previously diagnosed were found to be with unsatisfactory glycemic control. Compared to the 1994 survey, there was a significant increase in the prevalence of diabetes by 31.5%. Increase in age, increase in body mass index, and having a family history of diabetes were associated with increased odds of diabetes and IFG. While the level of education had no effect on IFG, higher level of education was associated with a decrease in the odds of having diabetes. CONCLUSION: The prevalence of type 2 diabetes and IFG is high in Jordan and is increasing. More than half of the patients with diabetes have unsatisfactory control. Therefore, they are likely to benefit from programs aimed at encouraging behaviors toward achieving optimum weight as well as physical activity behaviors. Physicians caring for patients with diabetes may need to adopt a more vigorous approach for diabetes control. PMID- 18413211 TI - Obesity and the use of insulin: a study of patients with type 2 diabetes in the UK. AB - OBJECTIVE: This analysis examines how obesity affects the prescribing of insulin for individuals with type 2 diabetes and poor glycemic control. METHODS: Data were obtained from the UK General Practice Research Database for the years 2000 2004. Patients were eligible if they had been identified as having type 2 diabetes and had undergone at least two valid glycosylated hemoglobin (HbA(1c)) tests. Additionally, patients had to have poor glycemic control on the index date (HbA(1c)>7.4), no use of insulin 6 months prior to the index date, and at least 30 months of data after the index date (N=6468). Descriptive statistics were used to examine unadjusted differences between obese and nonobese patients. A Cox proportional hazards model was applied to examine the relationship between obesity and the relative likelihood of initiation of insulin while controlling for differences in patient characteristics, medication use, and HbA(1c) levels. RESULTS: Obese individuals were significantly younger (P<.01), significantly more likely to be treated with two oral antidiabetic agents (P=.02), and significantly less likely to be treated with oral monotherapy (P=.02). Controlling for a wide range of factors that may impact receipt of insulin, results revealed that obese individuals had a "hazard" of initiation of insulin significantly lower than that of nonobese patients (hazard ratio=0.814, P=.01). CONCLUSIONS: Patient age, severity of illness, and prior medication use all affect whether the individual will be initially prescribed insulin. Moreover, the results of this study demonstrate that obesity is an additional critical factor in physicians' decision to begin treatment with insulin. PMID- 18413212 TI - TNF-alpha is an independent serum marker for proliferative retinopathy in type 1 diabetic patients. AB - PURPOSE: This study aimed to determine if there are any associations between serum levels of inflammatory markers and proliferative retinopathy (PDR) in type 1 diabetic patients. DESIGN: A cross-sectional design was utilized for this study. METHODS: One hundred twenty-eight type 1 diabetic patients underwent stereo fundus photography according to the Early Treatment Diabetic Retinopathy Study and were divided into two retinopathy groups: no or nonproliferative retinopathy (NDR/NPDR; n=62) and PDR (n=66). Serum levels of tumor necrosis factor-alpha (TNF-alpha), interleukin (IL)-1 beta, IL-6, soluble vascular cellular adhesion molecule-1 (sVCAM-1), soluble intercellular adhesion molecule-1 (sICAM-1), P-selectin, and high-sensitivity C-reactive protein (hsCRP) were analyzed. Statistical analysis was performed using nonparametric Mann-Whitney U test and multivariate logistic regression analysis. RESULTS: Patients with PDR had higher levels of TNF-alpha [7.0 pg/ml (<4-17) vs. 6.0 pg/ml (<4-25); P=.009], sVCAM-1 [860 ng/ml (360-2120) vs. 700 ng/ml (310-1820); P<.001], and P-selectin [180 ng/ml (39-400) vs. 150 ng/ml (42-440); P=.017; figures are expressed as median (range)]. There were no differences in serum levels of sICAM-1 or hsCRP. IL-1 beta was not detectable in any patient, and IL-6 was detectable in only 22.7% of the patients. In multivariate logistic regression analysis, TNF-alpha was the single, persistent, independent determinant inflammatory marker for PDR. CONCLUSION: The association between TNF-alpha and PDR in type 1 diabetic patients suggests that inflammation might play a role in the pathogenesis of proliferative diabetic retinopathy. PMID- 18413213 TI - Measurements of sweat response and skin vasomotor reflex for assessment of autonomic dysfunction in patients with diabetes. AB - AIMS: Some physical or arousal stimuli induce a rise in sweat secretion (sympathetic sweat response or SSwR) and a reduction in skin blood flow (skin vasomotor reflex or SkVR) to the palm. We recorded SSwRs and SkVRs in diabetic patients and assessed the usefulness of these parameters for evaluating autonomic dysfunction in diabetes. METHODS: We studied 42 diabetic patients (58+/-12 years) and 42 normal control subjects (59+/-11 years). Focal sweat secretion and skin blood flow were measured on the palm by a sudorometer and a Doppler flowmeter, respectively. SSwRs and SkVRs to deep inspiration, mental arithmetic, and isotonic exercise were recorded. SSwR amplitude was measured from baseline to peak, and SkVR amplitude (reduction rate) was calculated as: (blood flow reduction/basal blood flow)x100%. We also conducted head-up tilt tests and R-R interval variation tests (coefficient of variation of R-R intervals or CV(R-R)). RESULTS: The SSwR or SkVR amplitudes in the diabetic group were significantly lower than those in the control group for any stimulus. CV(R-R) in the diabetic group was significantly less than that in the control group. The diabetic group showed a significantly greater reduction in systolic blood pressure during head up tilt compared with the control group. In the diabetic group, there were significant correlations in SSwR or SkVR amplitudes versus blood pressure falls during the head-up tilt test, and CV(R-R) values. CONCLUSION: We believe that SSwR and SkVR are useful indexes for the evaluation of autonomic involvement in diabetic patients. PMID- 18413214 TI - The effect of venlafaxine HCl on painful peripheral diabetic neuropathy in patients with type 2 diabetes mellitus. AB - OBJECTIVE: The objective of this study was to evaluate the efficacy of venlafaxine HCl in the symptomatic treatment of painful peripheral diabetic neuropathy (PPDN) among patients with type 2 diabetes mellitus (DM). DESIGN: This study was designed as a prospective, randomized, and controlled trial. SETTING: This study was conducted at the Dicle University Medical Faculty (Diyarbakir, Turkey). PATIENTS: Sixty type 2 DM outpatients (47 females and 13 males) with PPDN who had a minimum visual analog scale (VAS) score of 40 mm were enrolled in this study. INTERVENTIONS: Patients randomized to the treatment group (n=30) received venlafaxine HCl, whereas those randomized to the control group (n=30) received a combination of vitamins B(1)and B(6) tablets. MEASURES: Severity of pain was measured by VAS, Short-Form McGill Pain Questionnaire, and numerical analog scale scores at admission and at the second, fourth, and eighth weeks of the study. Polyneuropathy was supported by electromyelography. OUTCOME: In the treatment group, severity of pain was measured as 70.0+/-13.0 in the VAS, as 24.9+/-6.2 in the Short-Form McGill Pain Questionnaire, and as 7.2+/-1.1 in the numerical analog scale. In the control group, it was measured as 73.0+/-8.0 in the VAS, as 26.8+/-6.2 in the Short-Form McGill Pain Questionnaire, and as 7.4+/ 0.8 in the numerical analog scale (P>.05). RESULTS: The most common form of PPDN was distal symmetrical sensorimotor polyneuropathy in both groups (46.8% vs. 50.0%). At the end of the study, there was a significant difference in severity of pain between the groups. In the treatment group, scores were 8.5+/-5.2 and 3.1+/-1.6 in the Short-Form McGill Pain Questionnaire and numerical analog scale, respectively; in the control group, these were 20.5+/-7.0 and 5.5+/-1.6, respectively (P<.001). CONCLUSIONS: Venlafaxine HCl is a safe and well-tolerable analgesic drug in the symptomatic treatment of PPDN; however, it has minimal adverse effects. It showed its efficacy markedly in the second week of therapy. PMID- 18413215 TI - A possible new risk factor in diabetic patients with erectile dysfunction: homocysteinemia. AB - INTRODUCTION AND OBJECTIVE: Increasing levels of homocysteine (Hcys) are related to decreased nitric oxide synthesis in the vascular endothelium and also to erectile dysfunction (ED). We investigated the traditional risk factors of ED and plasma levels of Hcys in diabetic patients. METHODS: Sixty-two men with type 2 diabetes and ages of between 40 and 70 years were included in the study. Detailed medical and sexual history and physical examination were performed. Erectile function was assessed with the International Index of Erectile Function. All participants were evaluated for fasting and postprandial plasma glucose, HbA1c, total cholesterol, triglyceride, high-density lipoprotein cholesterol, low density lipoprotein cholesterol, apolipoprotein A, apolipoprotein B-100, vitamin B(12), folic acid, and plasma Hcys levels. Patients were evaluated for diabetic neuropathy. Colored penile Doppler ultrasonography was used to detect vascular abnormalities in diabetic patients with ED. RESULTS: Fasting plasma glucose, postprandial plasma glucose, and HbA1c levels were significantly higher in diabetic patients with ED (P<.05). In addition, Hcys levels were slightly high in diabetic patients with ED (P=.079). Logistic regression analysis revealed that high levels of HbA1c (P<.01) and Hcys (P<.05), and smoking status (P<.01) were the main determinants of the presence of ED in our diabetic population. CONCLUSION: We conclude that traditional risk factors, poor metabolic control, and smoking are important in ED generation in diabetic patients. In addition, Hcys, which is an important risk factor in atherogenesis, seems to be important in ED generation in diabetic patients. PMID- 18413216 TI - Placental weight and placental weight-to-birth weight ratio are increased in diet and exercise-treated gestational diabetes mellitus subjects but not in subjects with one abnormal value on 100-g oral glucose tolerance test. AB - The aim of the present study was to determine whether the placental weight and placental weight-to-birth weight ratio (PW/BW) increased in pregnant women with one abnormal value (OAV) on 100-g oral glucose tolerance test (OGTT) and diet- and exercise-treated, non-insulin-requiring gestational diabetes mellitus (GDM) subjects. The 50-g glucose challenge test (GCT) was administered to 324 pregnant women. Women with abnormal 50-g test received a 100-g, 3-h OGTT using National Diabetes Data Group criteria. Women with GDM and OAV were treated with diet and exercise. Twenty subjects who required insulin or met exclusion criteria were excluded from the study. After the exclusion of 20 subjects, the GDM group consisted of 30 (9.7%) pregnant women and the OAV group consisted of 32 (9.9%) pregnant women. The control group consisted of 242 pregnant women. Birth weight (GDM: 3288.3+/-364.2 g; OAV: 3278.1+/-409.9 g; control group: 3270.6+/-346.5 g) did not differ significantly between groups (P>.05). Significantly higher placental weights (GDM: 694.8+/-152.1 g; OAV: 622.2+/-105.3 g; control group: 610.2+/-116.6 g; P<.01) and PW/BW (GDM: 0.21+/-0.03; OAV: 0.193+/-0.04; control group: 0.188+/-0.04; P<.01) were observed in GDM group compared to OAV and control group. No significant difference was found for OAV group in terms of placental weight and PW/BW compared to the control group. Our data indicated that women with OAV delivered infants and placenta of similar weight to those of normal pregnancies. PMID- 18413217 TI - Different accelerators to early-onset Type 2 diabetes: a comparison of Anglo Celtic and Chinese patients. AB - AIMS: The "accelerator hypothesis" postulates that metabolic syndrome (MS) factors-overweight and insulin resistance-increase functional demand on islets, accelerating diabetes onset to a younger age in both Type 1 and Type 2 diabetes (T2DM). Previous research has focused only on the former. We examine to what extent the MS and individual components are accelerators to the earlier onset of T2DM in Anglo-Celtic and Chinese populations. METHODS: A cross-sectional study of 1016 Anglo-Celtic and 1514 Chinese patients with recent-onset diabetes (duration <2 years) evaluated over a 12-year period. The MS syndrome and components were analyzed after stratification by age at presentation. RESULTS: The Anglo-Celtic group shows a high prevalence of MS in early-onset disease and a striking inverse relationship of body mass index (BMI) with age at presentation. For every increase in BMI of 1 kg/m(2), there is a reduction in the age of presentation by 0.5 years (r=-0.3; P<.0001) .Younger groups had a higher prevalence of insulin resistance, elevated triglyceride (Tg), and low high-density lipoprotein cholesterol (HDL-C) (P<.0001 for trend for all three indices). In contrast, the Chinese group showed no relationship between age of presentation with BMI, insulin resistance, Tg, or HDL-C. CONCLUSIONS: MS factors are important accelerators for T2DM in the Anglo-Celtic but not the Chinese population. This suggests that earlier onset of pancreatic beta cell deficiency is more important as an accelerator of diabetes presentation in Chinese. These data confirm the heterogeneity of T2DM and support the need for more ethnic specific strategies in diabetes prevention. PMID- 18413219 TI - Predictors of development and progression of microvascular complications in a cohort of Brazilian type 2 diabetic patients. AB - AIMS: Microvascular complications are associated with increased mortality in diabetes. The objective of this study was to investigate the predictors of microvascular complication development and progression in a prospective study of Brazilian type 2 diabetic patients. METHODS: A prospective follow-up study was carried out with 471 type 2 diabetic outpatients. Primary end points were the development or progression of retinopathy, peripheral neuropathy, and clinical nephropathy. Predictors were assessed for each individual microvascular complication and also as a composite outcome by Kaplan-Meier estimation of survival curves and by uni- and multivariate Cox analysis. RESULTS: During a median follow-up of 57 months (range 2-84 months), 196 patients (41.6%) developed or had a progression in microvascular disease. Retinopathy occurred in 22.5%, nephropathy in 19.1%, and neuropathy in 15.5% of the patients. In Cox multivariate analysis, increased echocardiographic left ventricular mass (LVM) and longer diabetes duration were selected as predictors for all end points. Higher mean fasting glycemia was a predictor for retinopathy and neuropathy, lower serum high-density lipoprotein (HDL) cholesterol for neuropathy, and higher total cholesterol for nephropathy. Increased LVM [hazard ratio (HR): 1.39, 95% CI: 1.23-1.56], higher fasting glycemia (HR: 1.19, 95% CI: 1.04-1.36), and longer diabetes duration (HR: 1.28, 95% CI: 1.11-1.47) were the predictors of the composite end point. CONCLUSIONS: Development and progression of microvascular complications in Brazilian type 2 diabetic patients are associated with worse hypertension and metabolic control. Additional studies are necessary to show if modification of these risk factors can reduce the burden of morbidity and mortality related to microvascular disease in type 2 diabetes. PMID- 18413218 TI - Cross-sectional associations of C-reactive protein with vascular risk factors and vascular complications in the DCCT/EDIC cohort. AB - OBJECTIVE: To determine the relationships between C-reactive protein (CRP) levels and features of Type 1 diabetes. RESEARCH DESIGN AND METHODS: Serum CRP was measured by nephelometry in a cross-sectional study of the Diabetes Control and Complications Trial/Epidemiology of Diabetes Interventions and Complications (DCCT/EDIC) cohort (n=983) and nondiabetic subjects (n=71). RESULTS: CRP levels [geometric mean (95% CI)] were higher in diabetic than in control subjects, 1.6 (1.5-1.7) vs. 1.2 (1.1-1.5) mg/l, P=.019. CRP was higher in diabetic women (n=438) than in men (n=545) [2.0 (1.8-2.3) vs. 1.3 (1.2-1.5), P<.001]. Diabetic subjects formerly in the DCCT intensive treatment group had higher CRP levels than those who were randomized to the conventional treatment group [1.8 (1.6 1.9), n=479 vs. 1.5 (1.3-1.6), n=456, P=.010], attributable to greater BMI in the prior intensive group. In diabetes, CRP correlated with HbA(1c) (r=0.13, P<.0001) and with insulin resistance traits: BMI (r=0.34, P<.0001), waist-to-hip ratio (WHR; males: r=0.35, P<.0001; females: r=0.22, P<.0001), diastolic blood pressure (r=0.07, P=.025), triglycerides (r=0.19, P<.0001), apoB (r=0.22, P<.0001), LDL particle concentration (r=0.26, P<.0001), and LDL particle size (r=-0.22, P<.0001). CRP was not associated with complications. Significant independent predictors of CRP in diabetes were gender, BMI, WHR, concurrent HbA(1c), and oral contraceptive pill use. CONCLUSIONS: CRP was elevated relative to nondiabetic subjects, and in diabetes was higher in females. Elevated CRP in Type 1 diabetes was associated with poor glycemic control, larger body habitus, and other factors that comprise the insulin resistance syndrome. Nevertheless, CRP levels were not associated with complications. Longitudinal studies are warranted. PMID- 18413220 TI - Renal effects of contrast media in diabetic patients undergoing diagnostic or interventional coronary angiography. AB - BACKGROUND: The use of safe iodinated contrast media (CM) to prevent contrast induced nephropathy (CIN) is an important consideration among renally impaired diabetic patients during coronary angiography. HYPOTHESIS: Diabetic patients with normal or mild renal dysfunction are less likely to receive renal protective measures during angiography, yet they may also be at risk for CIN. We compared the renal effects of iopamidol and iodixanol in diabetic patients who were referred for angiography. METHODS: Diabetic patients (N=122) with a serum creatinine (SCr) level of < or = 2 mg/dl were double-blind randomized to receive nonionic CM: iopamidol-370 (low osmolar, monomeric) or iodixanol-320 (iso osmolar, dimeric). Renal stability was evaluated at baseline and at Days 1, 3, and 7 post-angiography. The primary endpoint was a > or = 25% increase in SCr. RESULTS: Seventeen (10 iopamidol, 7 iodixanol; P=NS) patients had an increase in SCr > or = 25% over baseline. Over all days, analysis revealed nonsignificant differences in the incidence of CIN between the two study groups regardless of how CIN was defined. CONCLUSIONS: Diabetic patients with normal or mild renal dysfunction are at risk for CIN. No significant difference in renal response was observed for these CM in this at-risk population. PMID- 18413221 TI - Significant thresholds for the 75-g oral glucose tolerance test in pregnancy. AB - The significant threshold values for the 75-g oral glucose tolerance test (oGTT) during pregnancy have yet to be conclusively determined. This study aimed to identify the risk significance of various set thresholds for the oGTT result. Women undergoing a 75-g oGTT during the third trimester of pregnancy were classified into three groups: mild gestational impaired glucose tolerance (GIGT; 2-h postload glucose, 8.0-8.5 mmol/l; n=75), moderate-severe GIGT (8.6-10.9 mmol/l; n=167), and GDM (> or =11.0 mmol/l; n=76). Outcome indicators of these three groups of women were compared to the parameters of the women with a presumed normal carbohydrate metabolism (n=12,185). The results show that with increasing oGTT thresholds, there was an increasing risk of maternal morbidity in the form of hypertensive disorders complicating pregnancy, as well as obstetric intervention such as induction of labor, cesarean delivery, and preterm delivery. The infant was also at increasing risk with increasing oGTT thresholds from respiratory distress, macrosomia, and associated shoulder dystocia. It would appear, therefore, that abnormal glucose tolerance in pregnancy, even as defined by the World Health Organization criteria, has proportionate risks to both mother and child. PMID- 18413222 TI - Angiotensinogen gene T235 variant: a marker for the development of persistent microalbuminuria in children and adolescents with type 1 diabetes mellitus. AB - AIM: We examined genetic polymorphisms in the renin-angiotensin system (RAS) coding for angiotensin I-converting enzyme (ACE) insertion/deletion (I/D) for angiotensinogen (AGT) M235T and angiotensin II receptor type 1 (AGTR1) A1166C as predictors for the development of microalbuminuria (MA) in children with type 1 diabetes mellitus (T1DM). METHODS: Four hundred fifty-three (215 males, 238 females) T1DM children [median (interquartile range): age, 16.7 years (13.9 18.3); diabetes duration, 6.9 years (3.3-10.8); age at diagnosis, 9.1 years (5.8 11.8)] were followed prospectively from diagnosis until the development of MA (two of three consecutive overnight urine samples with albumin excretion rates of > or =20 and <200 microg/min). Kaplan-Meier survival curves and Cox proportional multivariate model estimated the probability of developing MA and the relative risk for MA among different variables. RESULTS: MA developed in 41 (9.1%) subjects. The frequencies of genotypes were as follows: ACE-II 112 (25%), ACE-ID 221 (49%), and ACE-DD 117 (26%) (n=450); AGT-MM 144 (32%), AGT-MT 231 (51%), and AGT-TT 77 (17%) (n=452); AGTR1-AA 211 (47%), AGTR1-AC 204 (45%), and AGTR1-CC 37 (8%) (n=452). The cumulative risk for the development of MA was higher in ACE-DD versus ACE-ID/II groups (log-rank test, P=.05), and a trend was noticed when AGT TT was compared to AGT-MT/MM groups (log-rank test, P=.08). AGT-TT polymorphism conferred a fourfold increased risk for MA compared to AGT-MM/MT (hazard ratio=3.8; 95% confidence interval=1.43-10.3; P=.008). INTERPRETATION: Our findings suggest that RAS gene polymorphism at AGT M235T is a strong predictor for early MA in young T1DM subjects. PMID- 18413223 TI - Influence of Lys656Asn polymorphism of the leptin receptor gene on insulin resistance in nondiabetic obese patients. AB - BACKGROUND: Alterations of the normal leptin receptor (LEPR) gene may be involved in the development of obesity. Leptin has been shown to be able to modulate insulin secretion. Different polymorphisms in the LEPR gene have been studied, albeit with unclear results. The polymorphism on codon 656 produces a change in charge, making this change possibly functional. OBJECTIVE: The objective of this study was to investigate the influence of Lys656Asn polymorphism in the LEPR gene on serum insulin, glucose values, and insulin resistance in the fasted state among obese men and women without diabetes mellitus. DESIGN: Two hundred thirty three (body mass index, >30 kg/m(2)) nondiabetic obese patients were analyzed. Indirect calorimetry, tetrapolar electrical bioimpedance, blood pressure determination, serial assessment of nutritional intake with 3-day written food records, and biochemical analysis were performed. Statistical analysis was performed for Lys656/Asn656 and Asn656/Asn656 jointly as a mutant allelic group and for Lys656/Lys656 as a wild allelic group. RESULTS: The subjects' (67 males and 166 females) mean age and mean body mass index were 43.6+/-16.6 years and 35.3+/-5.6 kg/m(2), respectively. One hundred forty-three patients (61.9%) had the genotype Lys656/Lys656 (wild group), whereas 88 (38.1%) had either the genotype Lys656/Asn656 (n=81; 30.7%) or the genotype Asn656/Asn656 (n=7; 7.4%) (mutant group). Age and sex distribution were similar in both groups. No difference was detected between the mutant and wild allelic groups in anthropometric parameters and dietary intakes. Homeostasis model assessment (HOMA; 2.8+/-1.7 vs. 5.6+/-4.8; P<.05) and insulin (18.1+/-10.7 vs. 32.1+/-25 mUI/ml; P<.05) levels were higher in males with the genotypes Lys656/Asn656 and Asn656/Asn656 than in males with the genotype Lys656/Lys656. Leptin levels were higher in males with a mutant genotype than in males with a wild genotype (39.3+/ 23 vs. 63.5+/-28 ng/ml; P<.05). CONCLUSION: The novel findings of our study are those of the association of the Lys656/Asn656 and Asn656/Asn656 genotypes with higher levels of insulin, HOMA, and leptin in males and the lack of such an association in females. PMID- 18413224 TI - Association of insulin sensitivity and glucose tolerance with the c.825C>T variant of the G protein beta-3 subunit gene. AB - The risk of macrovascular complications of diabetes mellitus is greatly enhanced by the presence of high blood pressure. In addition, hypertension and diabetes share insulin resistance as a common pathophysiological mechanism. Despite evidence for a common molecular genetic background of insulin resistance, glucose intolerance, and hypertension, few candidate genes have been shown to influence all of these features simultaneously. We examined the association of insulin sensitivity with the c.825C>T variant of the g-protein beta-3 subunit (GNB3), a candidate gene of hypertension, in families of Mexican-American hypertensive patients. METHODS: One hundred eighty subjects enrolled in a family study of Mexican-American hypertensive patients were recruited from hypertension clinics in Los Angeles. Subjects underwent pretreatment blood pressure recording, an oral glucose tolerance test, euglycemic hyperinsulinemic clamp, and anthropometric measurements. DNA from peripheral blood leukocytes was genotyped by polymerase chain reaction and restriction enzyme digest with BseD1 (GNB). Statistical analysis was performed by transmission disequilibrium testing. RESULTS: In carriers of the T-allele, blood glucose was significantly lower [(mean+S.D.) fasting: 96.7+22.9 vs. 106.7+51.7mg/dl, P=.009; oral glucose tolerance test (oGTT) 120 min: 131.7+48.7 vs. 137.8+64.9 mg/dl, P=.036], and insulin sensitivity was significantly higher (229.0+108.7 vs. 188.5+94.2 mg/kg per minute, P=.037) than in homozygous carriers of the C-allele. Blood pressure did not differ significantly between the phenotypes. CONCLUSION: In a Mexican-American hypertensive population, we found evidence for higher insulin sensitivity in carriers of the T allele of the c.825C>T variant of GNB3. PMID- 18413225 TI - Segmental blood flow and rheological determinants in diabetic patients with peripheral occlusive arterial disease. AB - Vascular abnormalities are more prevalent in the lower extremities in diabetic patients and may cause diminished perfusion to surrounding tissues. We sought to identify blood flow abnormalities in the leg of diabetic patients with peripheral occlusive arterial disease (POAD) and to determine whether these were associated with abnormalities in rheological determinants, namely, plasma fibrinogen concentration (PFC), relative plasma viscosity (RPV), hematocrit (Hct), and whole blood viscosity (WBV). Seventeen diabetic patients with POAD were compared with 40 diabetic patients without POAD and 19 nondiabetic control subjects. Blood flow was measured by venous occlusion plethysmography, RPV was measured by capillary viscometry, WBV was measured by a Wells-Brooksfield viscometer [at high (230 s( 1)) and low (23 s(-1)) shear rates], and PFC was measured by the clot-weight method of Ingram [Ingram, G. I. C. (1961). A suggested schedule for the rapid investigation of acute haemostatic failure. Journal of Clinical Pathology, 14, 356-360]. Ankle blood flow (Q(ak)) was significantly lower in diabetic patients with POAD than in diabetic patients without POAD (P<.05). PFC was higher and Hct was lower in diabetic patients with POAD than in diabetic patients without POAD (P<.05). RPV was 1.97+/-0.15 versus 1.92+/-0.15 in diabetic patients with POAD and diabetic patients without POAD, respectively (P>.05). There was no significant difference in WBV at low or high shear rates between the groups studied. There was a correlation between WBV at low shear rate and arterial flow in the calf (Q(c)) (r=.94) and great toe (r=.95) in diabetic patients with POAD, and between Q(c) and WBV at high (r=-.465) and low (r=-.472) shear rates in diabetic patients without POAD (P<.05). We conclude that vasodilatation occurring in diabetic patients without POAD is severely restricted or absent in diabetic patients with POAD. Increased plasma fibrinogen and plasma viscosity may contribute to this phenomenon. PMID- 18413226 TI - Predictors of quality of life among adults with type 2 diabetes mellitus. AB - Using a path model, this study examined the relationships among psychosocial factors (social support and acceptance of diabetes), knowledge of the disease, perceived difficulty of self-care behaviors (SCBs), and disease outcome [quality of life (QoL)] among 180 adults with type 2 diabetes mellitus. In general, respondents exhibited high levels of acceptance. The hypothesis that higher levels of social support and acceptance lead to lower perceived difficulty with SCBs was supported. High levels of social support increased acceptance and reduced SCBs. Knowledge reduced SCBs but was not associated with QoL. In the path model, SCBs influenced QoL and mediated the relationship between social support, acceptance, and QoL. PMID- 18413227 TI - Aspirin resistance associated with HbA1c and obesity in diabetic patients. AB - INTRODUCTION: Diabetes is known to be a prothrombotic state. Since serotonin uptake plays a role in both platelet activation and depression, we undertook to examine a hypothesis that aspirin resistance (AR) may be associated with both HbA1c and depressive symptoms and to assess other potential determinants of AR in diabetic patients. METHODS: A whole-blood desktop platelet function analyzer (PFA 100) with an epinephrine agonist was used to assess AR among patients with type 2 diabetes. AR was defined as PFA closure times <192 s. Depression symptoms were assessed with the Physicians Health Questionnaire. Patients being treated for type 2 diabetes (N=48) who took aspirin within the past 24 h constituted the study sample. Associations with AR were assessed with the use of the Mann-Whitney test and Fisher's Exact Test as well as with logistic regression models. RESULTS: AR was observed in 11 patients (23%) and was not significantly associated with age, sex, or race. AR was significantly associated with HbA1c > or = 8% (P=.002) and obesity (BMI> or = 30 kg/m(2); P=.01) and borderline associated with having > or = 1 depressive symptom (P=.07). Results were similar after multivariable adjustment in logistic regression models. No statistically significant associations of AR with age, sex, race, plasma glucose, blood pressure, cholesterol, or smoking were observed. CONCLUSION: These data suggest that AR may be of special concern for diabetic patients with poor glucose control and obesity. Whether the PFA-100 or any other practical measure of AR can be used in clinical practice to identify added cardiovascular disease risk and to inform platelet inhibition therapy needs further study. PMID- 18413229 TI - Engineering of protease variants exhibiting altered substrate specificity. AB - By using an improved genetic screening system, variants of the HAV 3CP protease which exhibit altered P2 specificity were obtained. We randomly mutated the His145, Lys146, Lys147, and Leu155 residues that constitute the S2 pocket of 3CP and then isolated variants that preferred substrates with Gln over the original Thr at the P2 position using a yeast-based screening method. One of the isolated variants cleaved the Gln-containing peptide substrate more efficiently in vitro, proving the efficiency of our method in isolating engineered proteases with desired substrate selectivity. PMID- 18413228 TI - A fluorescence polarization assay for identifying ligands that bind to vascular endothelial growth factor. AB - Vascular endothelial growth factor (VEGF) is a homodimeric proangiogenic protein that induces endothelial cell migration and proliferation primarily through interactions with its major receptors, VEGFR-1 and VEGFR-2. Inhibitors of one or both of these VEGF-receptor interactions could be beneficial as therapeutics for diseases caused by dysfunctional angiogenesis (e.g., cancer). Others have reported small peptides that bind to the VEGF dimer at surface regions that are recognized by the receptors. Here we report the development of a fluorescence polarization assay based on the binding to VEGF of a derivative of one of these peptides that has been labeled with BODIPY-tetramethylrhodamine (BODIPY(TMR)). This 384-well format assay is tolerant to dimethyl sulfoxide (DMSO, up to 4% [v/v]) and has a Z' factor of 0.76, making it useful for identifying molecules that associate with the receptor-binding surface of the VEGF dimer. PMID- 18413230 TI - Transient HMGB protein interactions with B-DNA duplexes and complexes. AB - HMGB proteins are abundant, non-histone proteins in eukaryotic chromatin. HMGB proteins contain one or two conserved "HMG boxes" and can be sequence-specific or nonspecific in their DNA binding. HMGB proteins cause strong DNA bending and bind preferentially to deformed DNAs. We wish to understand how HMGB proteins increase the apparent flexibility of non-distorted B-form DNA. We test the hypothesis that HMGB proteins bind transiently, creating an ensemble of distorted DNAs with rapidly interconverting conformations. We show that binding of B-form DNA by HMGB proteins is both weak and transient under conditions where DNA cyclization is strongly enhanced. We also detect novel complexes in which HMGB proteins simultaneously bind more than one DNA duplex. PMID- 18413231 TI - ERK activation is only one role of PKC in TCR-independent cytotoxic T cell granule exocytosis. AB - Cytotoxic T cells (CTLs) kill target cells by releasing lytic agents via regulated exocytosis. Three signals are known to be required for exocytosis: an increase in intracellular Ca(2+), activation of protein kinase C (PKC) and activation of extracellular signal regulated signal kinase (ERK). ERK activation required for exocytosis depends on activity of PKC. The simplest possibility is that the sole effect of PKC required for exocytosis is ERK activation. Testing this requires dissociating ERK and PKC activation. We did this using TCR independent stimulation of TALL-104 human leukemic CTLs. When cells are stimulated with thapsigargin and PMA, agents that increase intracellular Ca(2+) and activate PKC, respectively, PKC-dependent ERK activation is required for lytic granule exocytosis. Expressing a constitutively active mutant MAP kinase kinase activates ERK independent of PKC. However, activating ERK without PKC does not support lytic granule exocytosis, indicating that there are multiple effects of PKC required for granule exocytosis. PMID- 18413232 TI - Plasma kallikrein is activated on dermatan sulfate and cleaves factor H. AB - When human plasma is applied to a dermatan sulfate column, amidase activity is detected in the bound fraction and complement factor H is cleaved [A. Saito, H. Munakata, Factor H is a dermatan sulfate-binding protein: identification of a dermatan sulfate-mediated protease that cleaves factor H, J. Biochem. 137 (2005) 225-233]. Here, the amidase-active fraction was purified by sequential gel filtration and hydroxyapatite chromatography, and the amidase-active protein was identified to be plasma kallikrein by mass spectrometry. The activation of plasma kallikrein was further investigated by Western blotting using plasma deficient in prekallikrein or coagulation factor Xll. The dermatan sulfate column-bound fraction of the prekallikrein- and factor Xll-deficient plasmas did not show any amidase activity and factor H remained intact. Addition of kallikrein, but not activated factor Xll, to factor H purified from plasma resulted in cleavage of factor H. Thus, dermatan sulfate induces contact activation and activates kallikrein-mediated cleavage of FH. PMID- 18413233 TI - Global and focused transcriptional profiling of small molecule aminopeptidase N inhibitor reveals its mechanism of angiogenesis inhibition. AB - We recently developed a specific small molecule inhibitor of aminopeptidase N (APN), named as HNSA, through a high throughput screening. In the present study, we investigated the major cellular phenotypes of HNSA in comparison with those of APN knock-down in human fibrosarcoma cells and the mechanism of angiogenesis inhibition by the compound using DNA microarray analyses. Global gene expression analyses showed that HNSA signatures are significantly correlated with those of APN knock-down in HT1080 cells, suggesting that APN is a primary target of HNSA in the cells. Using the angiogenesis-focused DNA microarrays, nine of angiogenesis-related genes were identified as crucial mediators of angiogenesis inhibition by HNSA. These data demonstrate that HNSA can be used as a valuable tool to decipher the APN function in angiogenesis. PMID- 18413234 TI - Galectin-3 regulates RasGRP4-mediated activation of N-Ras and H-Ras. AB - Galectin-3 (Gal-3) is a pleiotropic beta-galactoside-binding protein expressed at relatively high levels in human neoplasms. Its carbohydrate recognition domain (CRD) contains a hydrophobic pocket that can accommodate the farnesyl moiety of K Ras. Binding of K-Ras to Gal-3 stabilizes K-Ras in its active (GTP-bound) state. Gal-3, which does not interact with N-Ras, was nevertheless shown to reduce N-Ras GTP in BT-549 cells by an unknown mechanism that we explored here. First, comparative analysis of various cancer cell lines (glioblastomas, breast cancer cells and ovarian carcinomas) showed a positive correlation between low N-Ras GTP/high K-Ras-GTP phenotype and Gal-3 expression levels. Next we found that epidermal growth factor-stimulated GTP loading of N-Ras, but not of K-Ras, is blocked in cells expressing high levels of Gal-3. Activation of Ras guanine nucleotide releasing proteins (RasGRPs) by phorbol 12-myristate 13-acetate (PMA) or downregulation of Gal-3 by Gal-3 shRNA increased the levels of N-Ras-GTP in Gal-3 expressing cells. We further show that the N-terminal domain of Gal-3 interacts with and inhibits RasGRP4-mediated GTP loading on N-Ras and H-Ras proteins. Growth of BT-549 cells stably expressing the Gal-3 N-terminal domain was strongly attenuated. Overall, these experiments demonstrate a new control mechanism of Ras activation in cancer cells whereby the Gal-3 N-terminal domain inhibits activation of N-Ras and H-Ras proteins. PMID- 18413235 TI - Bioimaging nitric oxide in activated macrophages in vitro and hepatic inflammation in vivo based on a copper-naphthoimidazol coordination compound. AB - Nitric oxide (NO) serves as a messenger for cellular signaling and physiological reactions such as inflammatory responses in vivo. Fluorescent bioimaging of nitric oxide is a very useful tool in NO functional research. Although many encouraging results have been achieved in the field of NO fluorescent detection, there is rarely satisfying result in inflammatory NO imaging in vivo. Here we report that fluorescent 5'-chloro-2-(2'-hydroxyphenyl)-1H-naphtho[2,3-d]imidazol can coordinate with Cu(II) to form a non-fluorescent coordination compound, which is able to directly and quickly image NO in cellular system or in vivo inflammation system with a turn-on fluorescence, based on a redox action of Cu(II). It was used to image NO produced by inducible nitric oxide synthase (iNOS) in lipopolysaccharide (LPS) activated murine macrophages. More importantly, it could image the NO production in an acute severe hepatic injury (ASHI) model of BALB/c mice induced by integrative LPS and D-galactosamine (GalN) treatment. The results prove that the 5'-chloro-2-(2'-hydroxyphenyl)-1H naphtho[2,3-d]imidazol coordinated with cupric ions can serve as an excellent NO bioimaging agent in different biological systems especially in inflammation related systems, and it may be valuable for diagnostic and pathological studies of NO related diseases. PMID- 18413236 TI - The kinetic properties producing the perfunctory pH profiles of catalase peroxidases. AB - Many structure-function relationship studies performed on the catalase-peroxidase enzymes are based on limited kinetic data. To provide a more substantive understanding of catalase-peroxidase function, we undertook a more exhaustive evaluation of catalase-peroxidase catalysis as a function of pH. Kinetic parameters across a broad pH range for the catalase and peroxidase activities of E. coli catalase peroxidase (KatG) were obtained, including the separate analysis of the oxidizing and reducing substrates of the peroxidase catalytic cycle. This investigation identified ABTS-dependent inhibition of peroxidase activity, particularly at low pH, unveiling that previously reported pH optima are clearly skewed. We show that turnover and efficiency of peroxidase activity increases with decreasing pH until the protein unfolds. The data also suggest that the catalase pH optimum is more complex than it is often assumed to be. The apparent optimum is in fact the intersection of the optimum for binding (7.00) and the optimum for activity (5.75). We also report the apparent pK(a)s for binding and catalysis of catalase activity as well as approximate values for certain peroxidatic and catalatic steps. PMID- 18413237 TI - Rab1b silencing using small interfering RNA for analysis of disease-specific function. AB - Rab1 GTPase is a critical component required for endoplasmic reticulum (ER)-to Golgi as well as intra-Golgi trafficking. It is required for the proper recruitment of tethering factors to mediate vesicle docking and subsequent fusion to the target membrane compartment. Much is known about the role of Rab1 in ER-to Golgi trafficking through overexpression of dominant negative mutation that inhibit GTP binding or GTPase activity of the protein, as well as through the use of antibodies to inhibit endogenous protein activity. These techniques have allowed for the establishment of a central role for Rab1 in trafficking in the early secretory pathway; however, the use of these techniques is limited. The introduction of antibodies relies on permeabilization of the cell membrane for their introduction. The use of dominant negative mutations relies on the mutation overwhelming the endogenous protein activity without removing it from the cell. The advent of siRNA to silence genes of interest provides a means to overcome this limitation. Here we describe optimal conditions for the efficient silencing of Rab1b using siRNA to analyze its role in disease. PMID- 18413238 TI - Rab8-optineurin-myosin VI: analysis of interactions and functions in the secretory pathway. AB - The small GTPase Rab8 has been shown to regulate polarized membrane trafficking pathways from the TGN to the cell surface. Optineurin is an effector protein of Rab8 and a binding partner of the actin-based motor protein myosin VI. We used various approaches to study the interactions between myosin VI and its binding partners and to analyze their role(s) in intracellular membrane trafficking pathways. In this chapter, we describe the use of the mammalian two-hybrid assay to demonstrate protein-protein interactions and to identify binding sites. We describe a secretion assay that was used in combination with RNA interference technology to analyze the function of myosin VI, optineurin, and Rab8 in exocytic membrane trafficking pathways. PMID- 18413239 TI - Characterization of Rab27a and JFC1 as constituents of the secretory machinery of prostate-specific antigen in prostate carcinoma cells. AB - Prostate-specific antigen (PSA) and prostate-specific acid phosphatase (PSAP) are produced by prostate carcinoma cells. Their secretion has implications in both prostate cancer diagnosis and progression. The mechanisms involved in PSA and PSAP secretion in response to androgens have remained relatively unknown. The small GTPase Rab27a regulates exocytosis in several tissues. Here, we present methods for the characterization of Rab27a and its effector JFC1/Slp1 as key components of the secretory machinery that regulates exocytosis in prostate carcinoma cells. PMID- 18413240 TI - In vitro assays to characterize inhibitors of the activation of small G proteins by their guanine nucleotide exchange factors. AB - Guanine nucleotide exchange factors (GEFs) are essential regulators of the spatiotemporal conditions of small GTP-binding protein (SMG) activation. Their cellular activities combine the biochemical stimulation of GDP/GTP exchange, which leads to the active conformation of the SMG, to the detection of upstream signals and, in some cases, interaction with downstream effectors. Inhibition of GEF activities by small molecules has become recently a very active field, both for understanding biology with the tools of chemistry and because GEFs are emerging as therapeutic targets. The natural compound brefeldin A (BFA) was the first inhibitor of a GEF to be characterized, and several inhibitors of SMG activation have since been discovered using a variety of screening methods. An essential step toward their use in basic research or as leads in therapeutics is the characterization of their mechanism of inhibition. GEFs function according to a multistep mechanism, involving transient ternary (nucleotide-bound) and binary (nucleotide-free) intermediates. This mechanism thereby offers many opportunities for blockage, but a thorough analysis is necessary to define the inhibition mechanism and the steps of the reaction that are affected by the inhibitor. Here, based on the case study of how BFA inhibits the activation of Arf activation by Sec7 domains, we describe a flowchart of assays to decipher the mechanism of inhibitors of the activation of SMGs by their GEFs. PMID- 18413241 TI - Analysis of small GTPase function in trypanosomes. AB - Trypanosomatids are protozoan parasites, of interest due to both their disease burden and deeply divergent position within the eukaryotic lineage. The African trypanosome, Trypanosoma brucei, has emerged as a very amenable model system, with a considerable toolbox of methods available, including inducible overexpression, RNA interference, and a completed genome. Here we describe some of the special considerations that need to be addressed when studying trypanosome gene function, and in particular small GTPases; we provide protocols for transfection, RNA interference, overexpression and basic transport assays, in addition to an overview of available vectors, cell lines, and strategies. PMID- 18413242 TI - Use of dynasore, the small molecule inhibitor of dynamin, in the regulation of endocytosis. AB - The large GTPase dynamin is essential for clathrin-dependent coated-vesicle formation. Dynasore is a cell-permeable small molecule that inhibits the GTPase activity of dynamin1, dynamin2 and Drp1, the mitochondrial dynamin. Dynasore was discovered in a screen of approximately 16,000 compounds for inhibitors of the dynamin2 GTPase. Dynasore is a noncompetitive inhibitor of dynamin GTPase activity and blocks dynamin-dependent endocytosis in cells, including neurons. It is fast acting (seconds) and its inhibitory effect in cells can be reversed by washout. Here we present a detailed synthesis protocol for dynasore, and describe a series of experiments used to analyze the inhibitory effects of dynasore on dynamin in vitro and to study the effects of dynasore on endocytosis in cells. PMID- 18413243 TI - Identification and verification of Sro7p as an effector of the Sec4p Rab GTPase. AB - Effectors are operationally defined as proteins that recognize a specific GTPase preferentially in its GTP-bound conformation. Here we present the use of affinity chromatography to identify potential effectors of Sec4p, the Rab GTPase that controls the final stage of the yeast secretory pathway. We describe the preparation of the Rab protein affinity matrix and the yeast lysate used in the purification. We also describe the methods used to identify and verify one candidate, Sro7p, as a bona fide Sec4p effector. This includes tests of the specificity and efficiency of binding both in vitro and in vivo. PMID- 18413244 TI - Characterization of Rab18, a lipid droplet-associated small GTPase. AB - Lipid droplets are the major intracellular store of lipids in eukaryotic cells. Understanding lipid storage and regulated mobilization of lipids from lipid droplets is essential for understanding the syndromes and diseases associated with excess lipid accumulation. Lipid droplets have been traditionally considered relatively inert structures. However, in recent years it has become apparent that lipid droplets are highly dynamic regulated organelles, which show complex interactions with other cellular compartments. The cellular components involved in regulation of lipid accumulation and release from lipid droplets, and in mediating the complex interactions with other organelles, are only now starting to be unraveled. A particularly important family of proteins in this respect is the Rab GTPases, crucial regulators of membrane traffic. Here we describe the techniques that we used to characterize the regulated association of Rab18 with the surface of lipid droplets. Rab18 provides an excellent marker to follow the dynamics of lipid droplets in living cells. In addition, the study of Rab18 provides insights into the mechanisms involved in the release of lipids from lipid droplets in adipocytes. In 3T3-L1 adipocytes, stimulation of lipolysis increases the association of Rab18 with lipid droplets, suggesting that recruitment of Rab18 is regulated by the metabolic state of individual lipid droplets. The study of Rab18 and its interacting proteins will provide new insights into the complex regulatory mechanisms involved in lipid storage and release. PMID- 18413245 TI - Analysis on the emerging role of Rab3 GTPase-activating protein in Warburg Micro and Martsolf syndrome. AB - Evidence is accumulating that Rab3A plays a key role in neurotransmitter release and synaptic plasticity. Recently mutations in the catalytic subunit p130 and the noncatalytic subunit p150 of Rab3 GTPase-activating protein were found to cause Warburg Micro syndrome and Martsolf syndrome, respectively, both of which exhibit mental retardation. We have found that loss of p130 in mice results in inhibition of Ca2+-dependent glutamate release from cerebrocortical synaptosomes and alters short-term plasticity in the hippocampal CA1 region, probably through the accumulation of the GTP-bound form of Rab3A. Here, we describe the procedures for the measurement of the GTP-bound pool of Rab3A with pull-down assay using mouse brains and the biochemical method for the measurement of glutamate release from mouse synaptosomes. PMID- 18413246 TI - Identification and characterization of JRAB/MICAL-L2, a junctional Rab13-binding protein. AB - The Rab family small G proteins are localized to distinct subsets of intracellular membranes and play a key role in membrane traffic through the interaction with their specific effector protein(s). Rab13 is identified as a plaque protein at tight junctions (TJs) and has been shown to regulate the assembly of functional TJs in epithelial cells. We have demonstrated that Rab13 mediates the endocytic recycling of integral TJ protein occludin, and identified a junctional Rab13-binding protein (JRAB)/molecule interacting with CasL-like 2 (MICAL-L2) as a Rab13 effector protein using a yeast two-hybrid system. JRAB/MICAL-L2 has a calponin-homology domain in the N-terminus, a LIM domain in the middle, and a coiled-coil domain at the C-terminus, and specifically binds to the GTP-bound form of Rab13 via its C-terminus. It is localized to TJs in epithelial cells and distributed along stress fibers in fibroblasts. In epithelial cells, JRAB/MICAL-L2 as well as Rab13 mediates the endocytic recycling of occludin, but not transferrin receptor, and the formation of functional TJs. This chapter describes the procedures for the isolation of JRAB/MICAL-L2 and the analysis of its functions. PMID- 18413247 TI - A SMAP gene family encoding ARF GTPase-activating proteins and its implication in membrane trafficking. AB - SMAP1 and SMAP2 proteins constitute a subfamily of the Arf-specific GTPase activating proteins. Both SMAP proteins bind to clathrin heavy chains and are involved in the trafficking of clathrin-coated vesicles. In cells, SMAP1 regulates Arf6-dependent endocytosis of transferrin receptors from the coated pits of the plasma membrane, whereas SMAP2 regulates Arf1-dependent retrograde transport of TGN38 from the early endosome to the trans-Golgi network. The common and distinct features of SMAP1 and SMAP2 activity provide a valuable opportunity to examine the differential regulation of membrane trafficking by these two proteins. In this chapter, we describe several basic experimental procedures that have been used to study the regulation of membrane trafficking using SMAP proteins, including a GAP assay as well as procedures to study the transport of transferrin receptors and TGN38. In addition, a yeast two-hybrid system is described because of its utility in identifying novel molecules that interact with SMAP. PMID- 18413248 TI - In vitro and in vivo analysis of neurotrophin-3 activation of Arf6 and Rac-1. AB - Arf GTP-binding proteins and Rho-family GTPases play key roles in regulating membrane remodeling and cytoskeletal reorganization involved in cell movement. Several studies have implicated neurotrophins and their receptors as upstream activators of these small GTP-binding proteins, however, the mechanisms and the cell type specificity of this neurotrophin activity are still under investigation. Here we describe the rationale and protocols used for the dissection of an NT3 activated pathway that leads to the specific activation of Arf6 and Rac1. PMID- 18413249 TI - Methods for analysis of rab27a/Munc13-4 in secretory lysosome release in hematopoietic cells. AB - Secretory lysosomes constitute a heterogeneous organelle of hematopoietic cells that combines the properties of regular lysosomes with those of secretory granules. Although secretory lysosomes serve essential functions, such as in the immune system and blood clotting, the mechanisms underlying the release of contents are incompletely understood. It is clear, however, that rab27a and the C2 domain protein munc13-4 serve essential functions. Mutations in these genes lead to immune disorders where the lytic granule function of cytotoxic T cells is jeopardized in humans. We identified munc13-4 as a rab27a binding protein from spleen. Munc13-4 is highly expressed in several hematopoietic cells including cytotoxic T cells and mast cells. We describe the molecular features of the interaction and requirements for localization, and show that munc13-4 is a positive regulator of secretory lysosome exocytosis. PMID- 18413250 TI - Analysis and expression of Rab38 in oculocutaneous lung disease. AB - Rab38 is a low-molecular-weight G-protein highly expressed in melanocytes of the skin and alveolar type II cells in the lung. A point mutation in the postulated GTP/GDP-interacting domain of Rab38 has been identified as the genetic lesion responsible for oculocutaneous albinism (OCA) in chocolate (cht) mice. Another point mutation that prevents translation of Rab38 mRNA is the molecular basis of the Ruby gene mutation causing the phenotype of OCA and prolonged bleeding time in Fawn-Hooded and Tester-Moriyama rats. Cht mice show conspicuously enlarged lamellar bodies in alveolar type II cells and abnormal lung structure. Triton X 114 phase partitioning of cht mouse lung showed that Rab38cht-protein was recovered in the aqueous phase. We produced recombinant Rab38cht-protein using a baculovirus/insect cell-protein expression system. The results demonstrate that Rab38cht-protein is inactive due to reduced membrane binding and enhanced intracellular degradation. Rab38 is a new strong candidate gene for human Hermansky-Pudlak syndrome (HPS) that is characterized by OCA, bleeding diathesis, and lung disease. PMID- 18413251 TI - Analysis of Rab1 function in cardiomyocyte growth. AB - Protein transport between intracellular organelles is coordinated by Rab GTPases. As an initial approach to defining the function of Rab GTPases in cardiomyocytes, our laboratory focused on Rab1, which regulates protein transport specifically from the endoplasmic reticulum (ER) to the Golgi apparatus. Our studies have demonstrated that adenovirus-driven expression of Rab1 promotes cell growth of primary cultures of neonatal cardiomyocytes in vitro and that transgenic expression of Rab1 in the myocardium induces cardiac hypertrophy in mouse hearts in vivo. These data provide strong evidence implicating that ER-to-Golgi protein transport functions as a regulatory site for control of cardiomyocyte growth. Here we describe a sets of methods used in our laboratory to characterize the function of Rab1 GTPase in modulating cardiac myocyte growth. PMID- 18413252 TI - Regulation of the trafficking and function of G protein-coupled receptors by Rab1 GTPase in cardiomyocytes. AB - G protein-coupled receptors (GPCRs) play a crucial role in regulating cardiac growth and function under normal and diseased conditions. It has been well documented that the precise function of GPCRs is controlled by intracellular trafficking of the receptors. Compared with the extensive studies on the events of the endocytic pathway, molecular mechanism underlying the transport process of GPCRs from the endoplasmic reticulum (ER) through the Golgi to the cell surface and regulation of receptor signaling by these processes in cardiac myocytes remain poorly defined. This chapter describes the methods to characterize the function of Rab1 GTPase, which modulates protein transport from the ER to the Golgi apparatus, in the trafficking and signaling of angiotensin II type 1 receptor (AT1R), alpha1-adrenergic receptor (AR), and beta-AR, and in hypertrophic growth in response to agonist stimulation in neonatal cardiac myocytes. PMID- 18413253 TI - Regulation of endosome dynamics by Rab5 and Huntingtin-HAP40 effector complex in physiological versus pathological conditions. AB - Vesicular transport of signaling molecules, specifically neurotrophins, in neurons is essential for their differentiation, survival, and plasticity. Neurotrophins such as neuron growth factor (NGF) and brain-derived neurotrophic factor (BDNF) are internalized by receptor-mediated endocytosis at synaptic terminals and loaded into endosomes for microtubule-based transport along axons to the cell body where they exert their signaling function in the nucleus. The molecular mechanisms underlying this intracellular transport are not only relevant from a basic knowledge viewpoint, but have also important implications for neurodegenerative diseases. Defects in trafficking are increasingly implicated in the pathology of Huntington's disease (HD) and other neurodegenerative disorders. The small GTPases Rab5 and Rab7 play important roles in the endocytic trafficking of neurotrophins. We have recently identified Huntingtin (Htt) and Huntingtin associated protein of 40 kDa (HAP40) as a novel Rab5 effector complex that regulates endosome motility. In HD, we detected higher HAP40 protein levels compared with normal cells. Such increase causes an augmented recruitment of Htt onto Rab5-positive early endosomes that drastically reduces their motility by "switching" these organelles from microtubules to F actin. These findings suggest a mechanism by which impaired Rab5-mediated trafficking of neurotrophic factors may be a key event of the pathogenetic process leading to neurodegeneration in HD. To dissect the mechanisms by which Htt, HAP40, and Rab5 function in early endosome interactions with the cytoskeleton, we developed assays to investigate endosome-cytoskeleton interactions that can be applied to normal and pathological conditions. We provide here detailed protocols for, first, an assay that measures binding of early endosomes to microtubules and F-actin. Second, we describe an improved protocol for a cell-free assay that recapitulates the motility of early endosomes along microtubules in vitro. These assays provide mechanistic insights into the dysfunction of endosome motility occurring in HD as well as other neurodegenerative disorders. PMID- 18413254 TI - Biochemical and biophysical analyses of Ras modification by ubiquitin. AB - Ras proteins are small GTPases that play key roles in the regulation of several cellular processes such as growth, differentiation, and transformation. Although Ras signaling was thought to occur uniformly on the inner leaflet of the plasma membrane, a growing body of evidence indicates that Ras activation happens dynamically within defined plasma membrane microdomains and at other specific intracellular compartments, thus ensuring the generation of distinct signal outputs. Yet the mechanisms that control the spatiotemporal segregation of Ras proteins remain poorly characterized. We have recently shown that the differential modification of Ras proteins by ubiquitination is a crucial factor that controls Ras intracellular trafficking and signaling potential. To better understand the process of Ras ubiquitination, it is important to establish assays that not only provide information about the nature of the ubiquitin modification involved, but also enable the monitoring of the dynamics of this process. In this chapter, we will describe biochemical and biophysical methodologies, namely immunoprecipitation, nickel-chelate affinity chromatography, and bioluminescence resonance energy transfer (BRET), for monitoring the ubiquitination of Ras proteins. Although the description focuses on Ras, the assays described can in principle be applied to the study of a range of proteins of interest that may be subject to ubiquitination, and the use of the different methods in parallel should provide new insights into the nature and dynamics of protein ubiquitination. PMID- 18413255 TI - Biochemical characterization of novel germline BRAF and MEK mutations in cardio facio-cutaneous syndrome. AB - Cardio-facio-cutaneous syndrome (CFC) is a sporadic, complex developmental disorder involving characteristic craniofacial features, cardiac defects, ectodermal abnormalities, growth deficiency, hypotonia, and developmental delay. CFC is caused by alteration of activity through the mitogen-activated protein kinase (MAPK) pathway due to heterogeneous de novo germline mutations in B-Raf mutant proteins, MEK1 and MEK2. Approximately 75% of individuals with CFC have mutations in BRAF. In vitro functional studies demonstrate that many of these mutations confer increase activity upon the mutant protein as compared to the wildtype protein. However, as is seen cancer, some of the B-Raf mutant proteins are kinase impaired. Western blot analyses corroborate kinase assays as determined by mutant proteins phosphorylating downstream effectors MEK and ERK. Approximately 25% of individuals with CFC have mutations in either MEK1 or MEK2 that lead to increased MEK kinase activity as judged by increased phosphorylation of its downstream effector ERK. Unlike BRAF, no somatic mutations have ever been identified in MEK genes. The identification of novel germline BRAF and MEK mutations in CFC will help understand the pathophysiology of this syndrome. Furthermore, it will also provide insight to the normal function of B-Raf and MEK, and contribute to the knowledge of the role of the MAPK pathway in cancer. Since the MAPK pathway has been studied intensively in the context of cancer, numerous therapeutics that specifically target this pathway may merit investigation in this population of patients. PMID- 18413256 TI - Biochemical and biological characterization of tumor-associated mutations of p110alpha. AB - Signaling by class I phosphatidylinositol 3-kinase (PI3K) controls cell growth, replication, motility, and metabolism. The PI3K pathway commonly shows gain of function in cancer. Two small GTPases, Rheb (Ras homolog enriched in brain) and Ras (rat sarcoma viral oncogene), play important roles in PI3K signaling. Rheb activates the TOR (target of rapamycin) kinase in a GTP-dependent manner; it links TOR to upstream signaling components, including the tuberous sclerosis complex (TSC) and Akt (homolog of the Akt8 murine lymphoma viral oncoprotein). Constitutively active, GTP-bound Rheb is oncogenic in cell culture, and activity that requires farnesylation. Ras activates PI3K by recruitment to the plasma membrane and possibly by inducing a conformational change in the catalytic subunit p110 of PI3K. In return, Ras signaling through the MAP kinase (MAPK) pathway is activated by PIP(3), the product of PI3K. Loss of Ras function can interfere with PI3K signaling. Various lines of evidence suggest complementary roles for PI3K and MAPK signaling in oncogenesis. PMID- 18413257 TI - Characterization of the Rheb-mTOR signaling pathway in mammalian cells: constitutive active mutants of Rheb and mTOR. AB - Rheb (Ras homolog enriched in brain) is a GTPase conserved from yeast to human and belongs to a unique family within the Ras superfamily of GTPases. Rheb plays critical roles in the activation of mTOR, a serine/threonine kinase that is involved in the activation of protein synthesis and growth. mTOR forms two distinct complexes, mTORC1 and mTORC2. While mTORC1 is implicated in the regulation of cell growth, proliferation, and cell size in response to amino acids and growth factors, mTORC2 is involved in actin organization. However, the mechanism of activation is not fully understood. Therefore, studies to elucidate the Rheb-mTOR signaling pathway are of great importance. Here we describe methods to characterize this pathway and to evaluate constitutive active mutants of Rheb and mTOR that we recently identified. Constitutive activity of the mutants can be demonstrated by the phosphorylation of ribosomal protein S6 kinase 1 (S6K1) and eIF4E-binding protein 1 (4E-BP1) both in vivo and in vitro after starving cells for amino acids and growth factors. In addition, formation and activity of mTORC1 and mTORC2 can be measured by immunoprecipitating these complexes and carrying out in vitro kinase assays. We also describe a protocol for rapamycin treatment, which directly inhibits mTOR and can be used to investigate the mTOR signaling pathway in cell growth, cell size, etc. PMID- 18413258 TI - Characterization of RalB-Sec5-TBK1 function in human oncogenesis. AB - The Ras-like GTPases, RalA and RalB, are key components of the oncogenic Ras signaling network. Recent evidence suggests that RalA and RalB collaborate to support tumorigenic transformation through distinct cell regulatory events. While RalA is apparently required to bypass normal restraints on cell proliferation, RalB is required to bypass normal restraints on cell survival. A direct Ral effector protein, Sec5, is a subunit of the exocyst complex, and is required to mediate RalB-dependent survival signals in transformed cells. Further analysis identified TBK1, a key mediator of the host defense response to viral challenge, as a novel Sec5 interacting protein essential for the capacity of RalB and Sec5 to deflect cell death in transformed cells. RalB activation promotes a direct interaction between Sec5 and TBK1 that results in TBK1 kinase activation via an unknown mechanism. Accordingly, both RalB and Sec5 are required for initiating host defense pathway activation upon virus infection. These observations revealed a novel relationship between molecular components of cell-autonomous innate immune signaling pathways and oncogenic transformation, and identified TBK1 as a potential target for therapeutic intervention in cancer. Here we describe details of methods, including protein complex analysis, protein kinase assays, host defense-response pathway activation, and cell transformation analysis, that can be used to investigate the contribution of the RalB-Sec5-TBK1 signaling cascade to both innate immune signaling and cell transformation. PMID- 18413259 TI - High-throughput lung cancer cell line screening for genotype-correlated sensitivity to an EGFR kinase inhibitor. AB - Human cancer cell lines that can be propagated and manipulated in culture have proven to be excellent models for studying many aspects of gene function in cancer. In addition, they can provide a powerful system for assessing the molecular determinants of sensitivity to anticancer drugs. They have also been used in recent studies to identify genomic alterations and gene expression patterns that provide important insights into the genetic features that distinguish the properties of tumor cells associated with similar histologies. We have established a large repository of human tumor cell lines (>1000) corresponding to a wide variety of tumor types, and we have developed a methodology for profiling the collection for sensitivity to putative anticancer compounds. The rationale for examining tumor cell lines on this relatively large scale reflects accumulating evidence indicating that there is substantial genetic heterogeneity among human tumor cells-even those derived from tumors of similar histologies. Thus, to develop an accurate picture of the molecular determinants of tumorigenesis and response to therapy, it is essential to study the nature of such heterogeneity in a relatively large sample set. Here, we describe the methodologies used to conduct such screens and we describe a "proof-of-concept" screen using the EGFR kinase inhibitor, erlotinib (Tarceva), with a panel of lung cancer lines to demonstrate a correlation between EGFR mutations and drug sensitivity. PMID- 18413260 TI - In vitro signaling by MAPK and NFkappaB pathways inhibited by Yersinia YopJ. AB - A procedure for an in vitro signaling assay is described for the MAPK and NFkappaB pathways. The method uses a membrane-cleared lysate that contains all the soluble components required for activating these signaling cascades. The pathways can be activated by variety of molecules, including kinases, G-proteins, and E3 ligases. We demonstrate that YopJ inhibits downstream of all these activators. The in vitro signaling assay is ideal for initial biochemical studies on activators and inhibitors of the MAPK and NFkappaB pathways. PMID- 18413261 TI - Biological and biochemical characterization of anthrax lethal factor, a proteolytic inhibitor of MEK signaling pathways. AB - The secretion of factors that block critical intracellular signaling pathways is a common strategy used by pathogenic bacteria for disabling host defenses and causing disease. Anthrax lethal toxin (LeTx) has been shown to cleave and inactivate mitogen-activated protein kinase (MAPK) kinases (MKKs or MEKs) and to inhibit MKK signaling. Cleavage of MKKs by LeTx prevents activation of their downstream substrates, the MAPKs. Because MAPK pathways regulate a variety of crucial cellular functions including proliferation, survival, differentiation, adhesion, and motility, LeTx has become a focus of study as an investigative tool as well as for the treatment and prevention of diseases due to malfunctions in MAPK signaling. This chapter describes methods for expressing and purifying the components of LeTx and focuses on techniques available for assessing its activity. PMID- 18413262 TI - Genetic analyses of the role of RCE1 in RAS membrane association and transformation. AB - Proteins terminating with a CAAX motif, such as the nuclear lamins and the RAS family of proteins, undergo post-translational modification of a carboxyl terminal cysteine with an isoprenyl lipid--a process called protein prenylation. After prenylation, the last three residues of CAAX proteins are clipped off by an endoprotease of the endoplasmic reticulum. RCE1 is responsible for the endoproteolytic processing of the RAS proteins and is likely responsible for endoproteolytic processing of the vast majority of CAAX proteins. Prenylation has been shown to be essential for the proper intracellular targeting and function of several CAAX proteins, but the physiologic importance of the endoprotease step has remained less certain. Here, we will review methods that have been used to define the physiologic importance of the endoproteolytic processing step of CAAX protein processing. PMID- 18413263 TI - Characterization of Kras-mediated pancreatic tumorigenesis in zebrafish. AB - Activating Kras mutations are a pervasive and characteristic feature of human pancreatic cancer. In order to examine the earliest in vivo effects of oncogenic Kras expression in the exocrine pancreas, we generated two lines of zebrafish expressing eGFP alone or eGFP fused to human Kras with an activating mutation in codon 12 (Kras G12V) driven by ptf1a regulatory elements using a BAC recombineering strategy (Park et al., 2008). In this review, we describe the techniques that we used to observe the effects of eGFP-Kras G12V expression in pancreatic progenitor cells of the zebrafish embryo, as well as techniques used to characterize malignant pancreatic tumors in the adult zebrafish. This zebrafish model of pancreatic neoplasia provides a unique view of the effects of oncogenic Kras in the embryonic pancreas and suggests that the zebrafish will be a useful model organism in which to study the biology of Kras-initiated pancreatic neoplasia. PMID- 18413264 TI - Genetic modeling of Ras-induced human rhabdomyosarcoma. AB - Rhabdomyosarcoma is the most common soft tissue sarcoma of childhood and adolescence. Historically, rhabdomyosarcoma has been studied by the manipulation of human cell lines derived from primary rhabdomyosarcoma tumor tissue adapted to grow in culture. Recently, mouse models have been added to the arsenal of tools to study this disease in vivo. However, given the emerging understanding of the genetic variability and mutability of human tumor-derived cell lines, and the existing differences between human and murine tumorigenesis, we sought to uniformly dissect the genetic events required to generate rhabdomyosarcoma from primary human skeletal muscle precursors. To this end, primary human skeletal muscle cells were transformed with defined genetic elements to corrupt the p53, Rb, Myc, telomerase, and Ras pathways, resulting in cells that, when assayed as subcutaneous xenografts in immunocompromised mice, formed tumors indistinguishable at the immunohistochemical level from the embryonal histologic variant of rhabdomyosarcoma. This chapter will discuss the techniques used to transform primary human skeletal muscle cells, the assays used to verify expression of the ectopically expressed genetic elements, and the methods used to evaluate the tumorigenic capacity of the resulting cell lines. PMID- 18413265 TI - Developmental processes and ectodermal contribution to the anal canal in mice. AB - The anorectal canal has two origins; the upper part is derived from endoderm and the lower part is derived from ectoderm. The process of ectodermal contribution to the canal remains unclear. To understand the development of this area, serial sagittal sections of mouse embryos were made every 12h from embryonic day 13.0 (E13.0) to E18.5. Three-dimensional (3-D) reconstructions were obtained from these sections. At the time of the disappearance of the cloacal membrane (E13.5), the endodermal lining reached the site of disintegrated membrane. Thus, the whole canal was of endodermal origin. The transitional zone between the dorsal end of the primary perineum and tail was thicker than other ectodermal epithelia. In this region, it changed from an acute to obtuse angle. After it straightened out and formed the canal, the secondary perineum appeared caudally. During these processes, the external sphincter appeared in the underlying mesenchyme of the thick ectoderm and functioned as a drawstring to form the ectodermal anal canal. PMID- 18413266 TI - Cementum thickness in multirooted human molars: a histometric study by light microscopy. AB - Cementum distribution was studied on transversely sectioned roots of 33 three rooted maxillary and 33 two-rooted mandibular permanent molars. The roots were sectioned at a level midway between furcation and apical region. Cementum thickness was measured at the midpoint of each of the four root surfaces and at a site exhibiting maximal cementum thickness. A reflected light microscope was used. Midpoint cementum thickness ranged between 5 and 800 microm in maxillary molars and between 5 and 700 microm in mandibular molars. Maximal cementum thickness ranged between 25 and 1140 microm in maxillary molars and between 20 and 700 microm in mandibular molars. The results indicate that cementum tends to accumulate along interradicular surfaces of multirooted molars. In addition, a tendency was noted for more cementum to occur orally than vestibularly in roots of mandibular molars as well as in buccal roots of maxillary molars. Furthermore, this study indicates that cementum accumulates in root concavities (developmental depressions), although the exact mechanism still awaits a clear explanation. Previous studies have suggested that tensile forces stimulate cementum apposition. It may be assumed, however, that masticatory function and continuing eruption of the teeth exert no discernible effect on cementum thickness in the examined region of the root. In conclusion, the observed cementum distribution could be attributed to at least two factors: one is buccal drift and the second may be the effect of root concavities. From a clinical viewpoint, the established pattern of cementum distribution in multirooted molars should be considered in endodontic and periodontal treatment. PMID- 18413267 TI - Mitigation of nociception via transganglionic degenerative atrophy: possible mechanism of vinpocetine-induced blockade of retrograde axoplasmic transport. AB - Vinpocetine, a derivative of vincamine, widely used in the clinical pharmacotherapy of cerebral circulatory diseases, inhibits retrograde axoplasmic transport of nerve growth factor (NGF) in the peripheral nerve, resulting in transganglionic degenerative atrophy (TDA) in the related ipsilateral superficial spinal dorsal horn, as shown in our previous publications. TDA induced by vinpocetine has been demonstrated to be followed by depletion of the marker enzyme fluoride-resistant acid phosphatase (FRAP) and its isoenzyme thiamine monophosphatase (TMP), and by the decrease in the pain-related neuropeptide substance P from laminae I-II-(III) from the segmentally related, ipsilateral substance of Rolando of the spinal cord. In the present paper, we report on the behavioral effects of perineurally administered vinpocetine. Nociception, induced by intraplantar injection of formalin, was mitigated by vinpocetine; increased expression of c-fos in the ipsilateral, segmentally related upper dorsal horn was also prevented. Since vinpocetine is not a microtubule inhibitor, and its chemical structure differs from that of vincristin and vinblastin (used formerly by us in the therapy of intractable, chronic neuropathic pain), its mode of action is enigmatic. We assume that the effect of vinpocetine in blocking retrograde axoplasmic transport of NGF might be related to its interaction with membrane trafficking proteins, such as signalling endosomes and the endocytosis mediating "pincher" protein. Temporary, locally restricted decrease of nociception, induced by vinpocetine, might be useful in the clinical treatment of intractable, chronic neuropathic pain, since vinpocetine can successfully be applied by transcutaneous iontophoresis. PMID- 18413268 TI - The topographic anatomy of the central artery: a neuronavigational-based study. AB - BACKGROUND/AIM: The central area of the brain, including the pre- and post central gyri with their vascular structures, is one of the most crucial regions to preserve in neurosurgical intervention. The central sulcus is also a landmark of utmost importance for neurosurgeons. Although the arteries supplying the central region have been briefly described, their exact course has not been studied. The aim of the study is to establish the position and course of the central sulcus artery in relation to the central sulcus and its convolutions. METHODS: Computer-assisted three-dimensional (3-D) anatomical reconstructions of the central area and of the central sulcus artery were performed with the aid of neuronavigational software in 13 patients operated at the Montreal Neurological Institute (MNI). RESULTS: The central sulcus artery was coincident with the central sulcus and course(s) was almost similar on both hemispheres. CONCLUSIONS: The knowledge of the exact course of the central sulcus artery is of paramount importance in cases of lesions or epileptic foci involving the central area. 3-D reconstruction with neuronavigation has proven to be a reproducible and reliable technique to provide the surgeon with the necessary surgical topographic anatomy of the central sulcus artery and central area. PMID- 18413269 TI - Innervation of pulmonary veins: morphologic pattern and pathways of nerves in the human fetus. AB - The aim of the study was to determine the anatomy of intrinsic nerves supplying human pulmonary veins (PVs). Twenty-two hearts of human fetuses with full sets of PVs were examined using a histochemical method for acetylcholinesterase in order to stain transmurally intrinsic neural structures on non-sectioned PVs for subsequent stereomicroscopic examination. Findings of the study demonstrate that epicardiac nerve extensions from both the dorsal right atrial and the middle dorsal subplexuses reached the right superior as well as the right inferior PVs, whereas the left superior PV was supplied by nerve extensions from the left dorsal subplexus. The left and middle dorsal subplexuses contributed nerves to the left inferior PV. The ganglia related topographically to PVs were patchy in distribution. On the left and right superior PVs, 38+/-6 and 31+/-3 ganglia were found, respectively, whereas 46+/-7 and 38+/-7 ganglia were identified on the left and right inferior PVs. The size of ganglia was similar for all four veins, ranging in area from 0.004+/-0.0003 to 0.007+/-0.0004 mm(2). The total area of ganglia distributed on a given PV was similar, ranging from 0.15+/-0.0003 to 0.25+/-0.0004 mm(2). The present findings demonstrate that the richest ganglion sites supplying intrinsic nerves to the human PVs are located on the posterior sides of both inferior and the left superior PVs and, therefore, these sites may be considered primary targets for focal pulmonary vein ablation in catheter-based therapy of atrial fibrillation. PMID- 18413270 TI - Effects of pyrophosphate on desmal and endochondral mineralization and TNAP activity in organoid culture. AB - During endochondral and desmal osteogenesis, mineralization of bone and cartilage matrix requires an appropriate solubility product of calcium and phosphate, collagen as a nucleator and deactivation of inhibitors, in order to prevent heterotopic calcification. In the 1960s, Fleisch and coworkers detected pyrophosphate (PPi) as an inhibitor of hydroxyapatite crystal growth, which should be removed by cleavage to tissue non-specific alkaline phosphatase (TNAP) activity. This theory had been established by basic experiments performed with collagen gels and demineralized matrices. In order to investigate the effect of PPi on matrix mineralization in bone and cartilage, calcium content and TNAP activity were measured in organoid cultures of mouse calvarial osteoblasts and limb bud cartilage after treatment with PPi and/or levamisole. In organoid cultures, bone and cartilage develop in a clear histotypical manner. PPi did not induce mineralization. Beta-glycerophosphate (beta-GP) and inorganic phosphate (Pi) induced mineralization which could be significantly reduced by PPi. Levamisole, an inhibitor of TNAP, also reduced mineralization; the combination with PPi was additive. TNAP activity was increased after treatment with PPi and levamisole in both osteoblast and cartilage cultures. Mineralization induced by beta-GP and Pi decreased TNAP activity in the osteoblast but not in cartilage organoid culture. In this culture system, PPi reduced mineralization as predicted by Fleisch's theory. Indications of cleavage of PPi were indirectly found because inhibition of hydrolysis of PPi by levamisole further reduced mineralization, probably due to the higher amounts of PPi available for binding to hydroxyapatite. PMID- 18413271 TI - Secondary induction and the development of tooth nerve supply. AB - During embryogenesis, dental trigeminal axon navigation and patterning in the developing tooth take place in a highly spatio-temporally directed manner that is tightly linked to tooth morphogenesis and cell differentiation. Tooth formation is regulated by sequential and reciprocal tissue interactions between dental epithelium and neural crest-derived ectomesenchymal cells. This odontogenic secondary induction is mediated by signal molecules of different conserved families. Recent molecular and experimental data have provided evidence that local instructive signaling from the early odontogenic epithelium also controls dental axon navigation in the dental mesenchyme. In this review, we discuss recent molecular data regarding tooth formation and innervation and the putative role of the secondary induction in coordinating these two developmental processes. Importantly, because it has not yet been shown that the interactions that regulate tooth innervation include signaling to the dental epithelium and that they are reciprocal, it remains to be demonstrated that secondary induction controls the establishment of tooth nerve supply. Moreover, the key question of which molecule(s), if any, integrate tooth morphogenesis and the development of dental sensory trigeminal innervation remains to be answered. PMID- 18413272 TI - Attention to self-referential stimuli: can I ignore my own face? AB - Auto-referential materials (i.e., the own name) have been described as particularly prone to capture attention. Some recent studies have questioned this view and shown that these own name effects are temporary and appear only in specific conditions: when enough resources are available (Harris, C. R., & Pashler, H. (2004). Attention and the processing of emotional words and names: Not so special after all. Psychological Science, 15, 171-178) or when the own name is presented within the focus of attention if it is a task-irrelevant stimulus (Gronau, N., Cohen, A., & Ben-Shakhar, G. (2003). Dissociations of personally significant and task-relevant distractors inside and outside the focus of attention: A combined behavioral and psychophysiological study. Journal of Experimental Psychology: General, 132, 512-529). In the present study, a stimulus that is unique to each individual was used: the self-face. In Experiment 1, the self-face produced a temporary distraction when presented at fixation during a digit-parity task. However, this distraction was not different from that triggered by another highly familiar face. In Experiment 2, the self-face failed to produce interference when presented outside the focus of attention. These results confirm recent findings showing that auto-referential materials do not automatically summon attention and have a distractive power only in specific conditions. PMID- 18413273 TI - Cannabis and the developing brain: insights from behavior. AB - The isolation and identification, in 1964, of delta-9-tetrahydrocannabinol (THC), the primary psychoactive compound in cannabis, opened the door to a whole new field of medical research. The exploration of the therapeutic potential of THC and other natural and synthetic cannabinoid compounds was paralleled by the discovery of the endocannabinoid system, comprising cannabinoid receptors and their endogenous ligands, which offered exciting new insights into brain function. Besides its well-known involvement in specific brain functions, such as control of movement, memory and emotions, the endocannabinoid system plays an important role in fundamental developmental processes such as cell proliferation, migration and differentiation. For this reason, changes in its activity during stages of high neuronal plasticity, such as the perinatal and the adolescent period, can have long-lasting neurobehavioral consequences. Here, we summarize human and animal studies examining the behavioral and neurobiological effects of in utero and adolescent exposure to cannabis. Since cannabis preparations are widely used and abused by young people, including pregnant women, understanding how cannabinoid compounds affect the developing brain, leading to neurobehavioral alterations or neuropsychiatric disorders later in life, is a serious health issue. In addition, since the endocannabinoid system is emerging as a novel therapeutic target for the treatment of several neuropsychiatric diseases, a detailed investigation of possible adverse effects of cannabinoid compounds on the central nervous system (CNS) of immature individuals is warranted. PMID- 18413274 TI - Giardia lamblia: biochemical characterization of an ecto-ATPase activity. AB - In this work, we describe the ability of living trophozoites of Giardia lamblia to hydrolyze extracellular ATP. In the absence of any divalent cations, a low level of ATP hydrolysis was observed (0.78+/-0.08 nmol Pi x h(-1)x10(-6) cells). The ATP hydrolysis was stimulated by MgCl(2) in a dose-dependent manner. Half maximum stimulation of ATP hydrolysis was obtained with 0.53+/-0.07 mM. ATP was the best substrate for this enzyme. The apparent K(m) for ATP was 0.21+/-0.04 mM. In the pH range from 5.6 to 8.4, in which cells were viable, this activity was not modified. The Mg(2+)-stimulated ATPase activity was insensitive to inhibitors of intracellular ATPases such as vanadate (P-ATPases), bafilomycin A(1) (V ATPases), and oligomycin (F-ATPases). Inhibitors of acid phosphatases (molybdate, vanadate and fluoride) or alkaline phosphatases (levamizole) had no effect on the ecto-ATPase activity. The impermeant agent DIDS and suramin, an antagonist of P2 purinoreceptors and inhibitor of some ecto-ATPases, decreased the enzymatic activity in a dose-dependent manner, confirming the external localization of this enzyme. Besides ATP, trophozoites were also able to hydrolyse ADP and 5 AMP, but the hydrolysis of these nucleotides was not stimulated by MgCl(2). Our results are indicative of the occurrence of a G. lamblia ecto-ATPase activity that may have a role in parasite physiology. PMID- 18413275 TI - Distribution of pigment dispersing hormone- and tachykinin-related peptides in the central nervous system of the copepod crustacean Calanus finmarchicus. AB - Peptides represent the largest class of signaling molecules used by nervous systems, functioning as locally-released paracrines and circulating hormones in both invertebrates and vertebrates. While many studies have focused on elucidating peptidergic systems in higher crustaceans, little is known about neuropeptides in the more primitive crustacean taxa. Here, we have begun an investigation of the peptides present in the central nervous system (CNS) of the copepod crustacean Calanus finmarchicus, presenting immunohistochemical data on the presence and distribution of pigment dispersing hormone (PDH) and tachykinin related peptide (TRP). In this species, strong PDH-like immunoreactivity was restricted to one pair of somata in the protocerebrum (PC) and the axonal projections emanating from them. TRP-like immunopositive structures were present in the PC, deutocerebrum (DC), tritocerebrum (TC), and ventral nerve cord (VNC). In the PC, a single soma in the left hemisphere was labeled. This neuron appears to be the source of a centrally located, bilaterally symmetric plexus present within the PC. In the DC, two pairs of intensely immunopositive somata were labeled, each projecting axons toward the posterior and producing an extensive collection of putative release terminals that spans the DC, TC, and anterior portion of the VNC. Several other more weakly labeled somata were also present in the DC. Double-labeling studies indicated that no co-localization of PDH- and TRP like peptides is present in the C. finmarchicus CNS. As preadsorption controls completely abolished each label, we feel these data represent accurate distributions of PDH- and TRP-like peptides within the C. finmarchicus CNS, thus providing a framework for future studies of the functional roles members of these peptide families play in this copepod species. PMID- 18413276 TI - Seasonal changes in imposex and tissue burden of butyltin compounds in Thais clavigera populations along the coastal area of Mirs Bay, China. AB - We assessed the current status of tributyltin (TBT) contamination of Thais clavigera (Gastropoda) along the coastal area of Mirs Bay, China for the first time. The snail samples were collected from 10 different sites in the summer (June) and winter (December) of 2006, respectively. They were analyzed for imposex status, i.e. relative penis size index (RPSI) and vas deferens sequence index (VDSI), followed by quantification of butyltins in their tissues. Most of the collected females suffered from imposex, but the level of butyltin contamination varied with the distance from Yantian Port, which is currently the fourth busiest container port in the world. The tissue concentration of TBT varied with season. For a particular site, the winter samples in general contained much higher concentration of TBT than the summer samples. RPSI, VDSI and organotin concentrations were higher in T. clavigera collected from sites closer to the Port, such as the Seafood Street and Kat O. Both RPSI and VDSI were positively correlated with the tissue burden of tributyltin. VDSI exhibited little seasonal variability, whereas RPSI showed marked seasonal variability, with lower values in the summer samples. The current results will serve as an important reference for long-term monitoring of butyltin contamination in this area. PMID- 18413277 TI - Evidence that exercise-induced heat storage is dependent on adrenomedullary secretion. AB - To investigate the influence of medullary adrenal secretion on thermoregulation during exercise, Phy (Eserine, 5x10(-3) M) was injected into the lateral cerebral ventricle of normal (INT) or bilaterally adrenodemedullated (ADM) untrained rats. Body temperature (Tb) and metabolic rate were measured in the rats while they were exercising on a treadmill (20 m min(-1), 5% inclination) until fatigue or while they were at rest after drug injection. In resting rats, Phy increased oxygen consumption in both INT or ADM rats without any effect on core temperature. During the dynamic phase of exercise (first 20 min), ADM attenuated the exercise-induced increase in core temperature (0.86+/-0.12 degrees C ADM Sal vs 1.48+/-0.21 degrees C INT Sal), thus reducing heat storage (HS) levels. Icv injection of Phy in ADM rats significantly reduced the increase in Tb (0.012+/ 0.10 degrees C min(-1) Phy vs 0.042+/-0.006 degrees C min(-1) Sal; p<0.02) and HS (65.8+/-56.1 cal Phy vs 207.7+/-32.7 cal Sal; p<0.04) compared to ADM Sal rats. In conclusion, the exercise-induced increase in heat storage was attenuated by adrenodemedullation in rats. Furthermore, the activation of heat loss mechanisms by the central cholinergic system during exercise occurs independently of adrenal medullary secretion suppression and can be improved by previous adrenodemedullation. Our data indicate the existence of a dual mechanism of heat loss control during the dynamic phase of exercise: one involving sympathoadrenal system activation that impairs heat loss and another that counteracts the increased sympathoadrenal activity through the hypothalamic cholinergic system to promote heat loss. PMID- 18413278 TI - Combating the "other diseases" of MDG 6: changing the paradigm to achieve equity and poverty reduction? AB - This paper suggests that the 'other diseases' of Millennium Development Goal 6 (MDG 6) are ignored by policy-makers and politicians who overfocus on unachievable objectives and targets around the 'big three' diseases of HIV, tuberculosis (TB) and malaria, which if the planet was viewed by aliens would be seen as the only diseases that existed on the planet. The diseases of the majority of the poor represent 'low hanging fruit' for control and elimination and opportunities are ignored despite the availability of cheap or donated drugs and ample evidence that such interventions are effective and reduce incidence, as well as mortality and morbidity. The time frame available to achieve the MDGs of some 7-8 years requires a re-evaluation of what can be done with the tools available now and which can address the problems faced by the majority of poor people afflicted by disabling conditions which together represent a global burden greater than malaria or TB. The author considers also the volume of research relevant to the MDGs and their achievement is distorted by the focus on high tech end research which cannot be delivered by 2015 and that in terms of the 90:10 gap in research relevant to the problems of the poorest the real gap is 99:1. The concepts of distortion of donor funding for diseases of MDG 6 for implementation of largely curative interventions which do not reduce incidence as well as research which addresses problems that cannot reach poor people in the time frame to 2015 is emphasised. New paradigms are required if any impact on MDG 6 is to be achieved recognising the needs of the majority via an equitable distribution of funding. PMID- 18413279 TI - Sorption kinetics and its effects on retention and leaching. AB - Sorption of pesticides to substrates used in biopurification systems is important as it controls the system's efficiency. Ideally, pesticide sorption should occur fast so that leaching of the pesticide in the biopurification system is minimized. Although modeling of pesticide transport commonly assumes equilibrium, this may not always be true in practice. Sorption kinetics have to be taken into account. This study investigated the batch sorption kinetics of linuron, isoproturon, metalaxyl, isoxaben and lenacil on substrates commonly used in a biopurification system, i.e. cow manure, straw, willow chopping, sandy loam soil, coconut chips, garden waste compost and peat mix. The first-order sorption kinetics model was fitted to the observed pesticide concentrations versus time resulting in an estimated kinetic rate constant alpha. Sorption appeared to be fast for the pesticides linuron and isoxaben, pesticides which were classified as immobile, while less mobile pesticides displayed an overall slower sorption. However, the substrate does not seem to be the main parameter influencing the sorption kinetics. Coconut chips, which is a substrate with a high organic matter content showed slow sorption for most of the pesticides. The effect of different estimated alpha values on the breakthrough of pesticides through a biopurification system was evaluated using the HYDRUS 1D model. Significant differences in leaching behavior were observed as a result of the obtained differences in sorption kinetics. PMID- 18413280 TI - Cyclodextrin-based supramolecular architectures: syntheses, structures, and applications for drug and gene delivery. AB - The supramolecular structures formed between cyclodextrins (CDs) and polymers have inspired interesting developments of novel supramolecular biomaterials. This review will update the recent progress in studies on supramolecular structures based on CDs and block copolymers, followed by the design and synthesis of CD based supramolecular hydrogels and biodegradable polyrotaxanes for potential controlled drug delivery, and CD-containing cationic polymers and cationic polyrotaxanes for gene delivery. Supramolecular hydrogels based on the self assembly of the inclusion complexes between CDs with biodegradable block copolymers could be used as promising injectable drug delivery systems for sustained controlled release of macromolecular drugs. Biodegradable polyrotaxanes with drug-conjugated CDs threaded on a polymer chain with degradable end-caps could be interesting supramolecular prodrugs for controlled and targeting delivery of drugs. CD-containing cationic polymers as gene carriers showed reduced cytotoxicity than non-CD-containing polymer counterparts. More importantly, the polyplexes of CD-containing cationic polymers with DNA could be pegylated through a supramolecular process using inclusion complexation between the CD moieties and a modified PEO. Finally, new cationic polyrotaxanes composed of multiple oligoethylenimine-grafted CDs threaded and end-capped on a block copolymer chain were designed and synthesized as a new class of polymeric gene delivery vectors, where the chain-interlocked cationic cyclic units formed an integrated supramolecular entity to function as a macromolecular gene vector. The development of the supramolecular biomaterials through inclusion complexation has opened up a new approach for designing novel drug and gene delivery systems, which may have many advantages over the systems based on the conventional polymeric materials. PMID- 18413281 TI - Combined action and regulation of phase II enzymes and multidrug resistance proteins in multidrug resistance in cancer. AB - A major limitation in the treatment of cancer patients is the ability of cancer cells to become resistant to chemotherapeutic drugs, a phenomenon known as multidrug resistance (MDR). Two important mechanisms involved in multidrug resistance are the increased activity of efflux pumps, such as those of the multidrug resistance proteins (MRPs) and the detoxification by phase II conjugating enzymes, like glutathione S-transferases and UDP glucuronosyltransferases. A synergistic interaction between these two mechanisms, MRPs and phase II enzymes, in conferring MDR has been shown for multiple anticancer drugs. In addition, there is substantial evidence of a coordinate regulation of the expression of phase II enzymes and MRPs, most likely mediated by the nuclear factor-erythroid 2 p45-related factor (Nrf2) and antioxidant response elements. Further elucidation of the combined action and regulation of phase II enzymes and MRPs in MDR will be an aid in the improvement of the chemotherapeutic treatment of cancer patients. PMID- 18413282 TI - Generalized comfort/placebo means should be used in cancer care. PMID- 18413283 TI - Methylene blue and iodine adsorption onto an activated desert plant. AB - Although frequently less toxic than many colorless effluents, colored effluents are generally considered by the public as an indicator of pollution. The present investigation aimed at identifying the effectiveness of a local desert plant characteristic of Southwest Algeria and known as Salsolavermiculata, which was pyrolyzed and treated chemically with a 50% zinc chloride solution, to remove methylene blue and iodine. The natural plant adsorption capacities were respectively 23mg/g and 272mg/g for methylene blue and iodine. Corresponding results for the pyrolyzed plant uptakes were 53mg/g and 951mg/g, while those for the pyrolyzed plant, chemically treated and activated at 650 degrees C, were 130mg/g and 1178mg/g, respectively. In comparison, the standard Merck activated carbon capacities were 200mg/g for methylene blue and 950mg/g for iodine. Consequently, this low-cost local plant may also prove useful for the removal of large organic molecules as well as potential inorganic contaminants. PMID- 18413285 TI - Biological testing of a digested sewage sludge and derived composts. AB - Aiming to evaluate a possible loss of soil habitat function after amendment with organic wastes, a digested sewage sludge and derived composts produced with green residues, where biologically tested in the laboratory using soil animals (Eisenia andrei and Folsomia candida) and plants (Brassica rapa and Avena sativa). Each waste was tested mimicking a field application of 6ton/ha or 12ton/ha. Avoidance tests did not reveal any impact of sludge and composts to soil biota. Germination and growth tests showed that application of composts were beneficial for both plants. Composts did not affect earthworm's mass increase or reproduction, but the highest sludge amendment revealed negative effects on both parameters. Only the amendment of composts at the highest dose originated an impairment of springtails reproductive output. We suggest that bioassays using different test species may be an additional tool to evaluate effects of amendment of organic wastes in soil. Biological tests are sensitive to pollutants at low concentrations and to interactions undetected by routine chemical analysis. PMID- 18413284 TI - Stability of agar in the seaweed Gracilaria eucheumatoides (Gracilariales, Rhodophyta) during postharvest storage. AB - The status of the cell-wall polysaccharide of the red seaweed, Gracilaria eucheumatoides upon postharvest storage was assessed in this study. The yield, chemical composition, physical and textural properties of alkali-treated agar extract was determined at different time intervals within 31 months of storage at dried state after harvest. Minimal fluctuation in agar yield was observed, ranging from 22.9% to 29.0%. The gel strength of agar extracts averaged 318gcm( 2) until the third month of storage but decreased considerably thereafter. The relative viscosity and molecular weight of the extracts varied inversely with storage time. Results indicated that both physical and textural parameters of agar generally decreased with storage time, likely due to depolymerization as indicated by decrease in molecular weight. Agar extracted from seaweeds up to 3 months of storage could be considered to exhibit gel quality suitable for food applications. Prolonged storage of the seaweed harvest is not recommended. PMID- 18413286 TI - Expression of thyroid transcription factor-1 in brain metastases: a useful indicator of pulmonary origin. AB - Determining the primary site of a brain metastasis can be difficult, particularly if there is no organ-specific tumor histology. Thyroid transcription factor-1 (TTF-1) is a marker commonly expressed in lung and thyroid carcinomas. We studied the value of TTF-1 staining to identify brain metastases from a lung primary. Forty-three consecutive patients metastatic carcinomas of the brain were reviewed from a 4 year period. Twenty-one (75%) of the 28 metastatic pulmonary carcinomas were TTF-1 positive (+), whereas only one metastatic papillary thyroid carcinoma (7%) out of 15 metastatic non-pulmonary carcinomas was TTF-1 (+). We also studied the cytokeratin profile of these tumors. However, cytokeratin profile, either alone or combined with TTF-1 staining, was not better than TTF-1 staining alone in identifying brain metastases of pulmonary origin. Overall, TTF-1 positivity is a useful indicator of pulmonary origin in brain metastases, with a very high positive predictive value (95%) and a moderate negative predictive value (67%). PMID- 18413287 TI - Nociceptin/orphanin FQ receptor blockade attenuates MPTP-induced parkinsonism. AB - Endogenous nociceptin/orphanin FQ (N/OFQ) inhibits the activity of dopamine neurons in the substantia nigra and affects motor behavior. In this study we investigated whether a N/OFQ receptor (NOP) antagonist, J-113397, can modify movement in naive mice and nonhuman primates and attenuate motor deficits in MPTP treated parkinsonian animals. J-113397 facilitated motor activity in naive mice at low doses (0.1-1 mg/kg) and inhibited it at higher ones (10 mg/kg). Likewise, in MPTP-treated mice, J-113397 reversed motor deficit at 0.01 mg/kg but worsened hypokinesia at higher doses (1 mg/kg). In naive nonhuman primates, J-113397, ineffective up to 1 mg/kg, produced inconsistent motor improvements at 3 mg/kg. Conversely, in parkinsonian primates J-113397 (0.01 mg/kg) reversed parkinsonism, being most effective against hypokinesia. We conclude that endogenous N/OFQ modulates motor activity in mice and nonhuman primates and contributes to parkinsonian symptoms in MPTP-treated animals. NOP receptor antagonists may represent a novel approach to Parkinson's disease. PMID- 18413288 TI - Autophagy and endocytosis in the amnion. AB - To search for the origin of nutrition in the amnion, we focused attention on both endocytotic and autophagic pathways. Using ultrastructural and biochemical methods, we examined 20 human amnions at term gestation. The uptake of horseradish peroxidase (HRP) was used for the detection of endocytosis. Transfection of the LC3-GFP plasmid and staining with monodansylcadaverine (MDC) and LysoTracker red (LTR) were used to demonstrate the formation of autophagic vacuoles. In addition, two autophagic genes, beclin 1 and Atg5, were assayed by RT-PCR. Within the amniotic epithelial (AE) cells, autophagic vacuoles contained organelles and cytoplasmic components and were enclosed by a double membrane. They contained autophagosomes with transfected LC3-GFP that stained positive for MDC and autolysosomes that stained positive for LTR. Endocytosis was an extremely active process in the cellular uptake of fluid and fluid contents and led to formation of vesicles and endosomes, which were found to be positive by HRP test. Many uniform vesicles were collected in the multivesicular bodies (MVBs). Finally, both endosomes and autophagosomes were fused and degraded by lysosomes. The data also demonstrated that large autophagosomes engulfed some endosomes or MVBs. Transcription of beclin 1 and Atg5 occurred in the amnion at term gestation. Taken together, these results show that AE cells have active endocytotic and autophagic capacities and that lysosomes are involved in the intracellular degradation of endosomes and autophagosomes. Sometimes the autophagic and endocytotic pathways converge. This study suggests that of endocytosis and autophagy activities in AE cells can be induced by nutrient limitation and are probably also evoked in response to some hormones in the amniotic fluid. Activation of both endocytotic and autophagic pathways plays different roles in the ability of the cell to acquire nutrients needed for its survival. PMID- 18413289 TI - Widespread reward-system activation in obese women in response to pictures of high-calorie foods. AB - Behavioral studies have suggested that exaggerated reactivity to food cues, especially those associated with high-calorie foods, may be a factor underlying obesity. This increased motivational potency of foods in obese individuals appears to be mediated in part by a hyperactive reward system. We used a Philips 3T magnet and fMRI to investigate activation of reward-system and associated brain structures in response to pictures of high-calorie and low-calorie foods in 12 obese compared to 12 normal-weight women. A regions of interest (ROI) analysis revealed that pictures of high-calorie foods produced significantly greater activation in the obese group compared to controls in medial and lateral orbitofrontal cortex, amygdala, nucleus accumbens/ventral striatum, medial prefrontal cortex, insula, anterior cingulate cortex, ventral pallidum, caudate, putamen, and hippocampus. For the contrast of high-calorie vs. low-calorie foods, the obese group also exhibited a larger difference than the controls did in all of the same regions of interest except for the putamen. Within-group contrasts revealed that pictures of high-calorie foods uniformly stimulated more activation than low-calorie foods did in the obese group. By contrast, in the control group, greater activation by high-calorie foods was seen only in dorsal caudate, whereas low-calorie foods were more effective than high-calorie foods in the lateral orbitofrontal cortex, medial prefrontal cortex, and anterior cingulate cortex. In summary, compared to normal-weight controls, obese women exhibited greater activation in response to pictures of high-calorie foods in a large number of regions hypothesized to mediate motivational effects of food cues. PMID- 18413291 TI - Phylogenetic relationships among sea anemones (Cnidaria: Anthozoa: Actiniaria). AB - Sea anemones (order Actiniaria) are among the most diverse and successful members of the anthozoan subclass Hexacorallia, being found at all depths and latitudes and in all marine habitats. Members of this group exhibit the greatest variation in anatomy, biology, and life history in Hexacorallia, and lack any morphological synapomorphy. Nonetheless, previous molecular phylogenetic studies have found that Actiniaria is monophyletic with respect to other extant hexacorallians. However, relationships within Actiniaria have remained unresolved, as none of these earlier works have included sufficient taxon sampling to estimate relationships within Actiniaria. We have analyzed sequences from two mitochondrial and two nuclear markers for representatives of approximately half of the family-level diversity within the order, and present the first phylogenetic tree for Actiniaria. We concur with previous studies that have suggested that molecular evolution is unusually slow in this group. We determine that taxonomic groups based on the absence of features tend not to be recovered as monophyletic, but that at least some classical anatomical features define monophyletic groups. PMID- 18413290 TI - Developmental changes in brain regions involved in phonological and orthographic processing during spoken language processing. AB - Developmental differences in brain activation of 9- to 15-year-old children were examined during an auditory rhyme decision task to spoken words using functional magnetic resonance imaging (fMRI). As a group, children showed activation in the left superior/middle temporal gyri (BA 22, 21), right middle temporal gyrus (BA 21), dorsal (BA 45, pars opercularis) and ventral (BA 46, pars triangularis) aspects of the left inferior frontal gyrus, and left fusiform gyrus (BA 37). There was a developmental increase in activation in the left middle temporal gyrus (BA 22) across all lexical conditions, suggesting that automatic semantic processing increases with age regardless of task demands. Activation in the left dorsal inferior frontal gyrus also showed developmental increases for the conflicting (e.g. PINT-MINT) compared to the non-conflicting (e.g. PRESS-LIST) non-rhyming conditions, indicating that this area becomes increasingly involved in strategic phonological processing in the face of conflicting orthographic and phonological representations. Left inferior temporal/fusiform gyrus (BA 37) activation was also greater for the conflicting (e.g. PINT-MINT) condition, and a developmental increase was found in the positive relationship between individuals' reaction time and activation in the left lingual/fusiform gyrus (BA 18) in this condition, indicating an age-related increase in the association between longer reaction times and greater visual-orthographic processing in this conflicting condition. These results suggest that orthographic processing is automatically engaged by children in a task that does not require access to orthographic information for correct performance, especially when orthographic and phonological representations conflict, and especially for longer response latencies in older children. PMID- 18413292 TI - New real-time PCR tests for species-specific detection of Chlamydophila psittaci and Chlamydophila abortus from tissue samples. AB - Chlamydophila psittaci and Chlamydophila abortus are the causative agents of avian chlamydiosis (psittacosis) and ovine enzootic abortion, respectively. Both pathogens are known to possess zoonotic potential. Due to their close genetic relatedness, direct and rapid species identification is difficult. In the present study, new real-time PCR assays are reported for both species. The tests are based on highly specific probes targeting the ompA gene region and were conducted as duplex PCRs including an internal amplification control. The Cp. psittaci assay successfully passed a proficiency test at national level. Examination of field samples revealed Cp. psittaci as the dominating species in birds, but also Cp. abortus in a few psittacines. Real-time PCR assays for species-specific detection of Cp. psittaci and Cp. abortus are suited for routine diagnosis, which renders them important tools for the recognition of outbreaks of psittacosis and ovine enzootic abortion. PMID- 18413293 TI - Methionine synthase reductase deficiency results in adverse reproductive outcomes and congenital heart defects in mice. AB - Low dietary folate and polymorphisms in genes of folate metabolism can influence risk for pregnancy complications and birth defects. Methionine synthase reductase (MTRR) is required for activation of methionine synthase, a folate- and vitamin B(12)-dependent enzyme. A polymorphism in MTRR (p.I22M), present in the homozygous state in 25% of many populations, may increase risk for neural tube defects. To examine the impact of MTRR deficiency on early development and congenital heart defects, we used mice harboring a gene-trapped (gt) allele in Mtrr. Female mice (Mtrr(+/+), Mtrr(+/gt), and Mtrr(gt/gt)) were mated with male Mtrr(+/g) mice. Reproductive outcomes and cardiac phenotype (presence of defects and myocardial thickness) were assessed at E14.5. Mtrr-deficient mothers had more resorptions and more delayed embryos per litter (resorptions per litter: 0.29+/ 0.13; 1.21+/-0.41; 1.87+/-0.38 and delayed embryos per litter: 0.07+/-0.07; 0.14+/-0.14; 0.60+/-0.24 in Mtrr(+/+), Mtrr(+/gt), and Mtrr(gt/gt) mothers respectively). Placentae of Mtrr(gt/gt) mothers were smaller and their embryos were smaller, with myocardial hypoplasia and a higher incidence of ventricular septal defects (VSD) per litter (0; 0.57+/-0.30; 1.57+/-0.67 in Mtrr(+/+), Mtrr(+/gt), and Mtrr(gt/gt) groups respectively). Embryonic Mtrr(gt/gt) genotype was associated with reduced embryonic length, reduced embryonic and placental weight, and higher incidence of VSD, but did not affect myocardial thickness or embryonic delay. We conclude that Mtrr deficiency adversely impacts reproductive outcomes and cardiac development in mice. These findings may have implications for nutritional prevention of heart defects, particularly in women with the common MTRR polymorphism. PMID- 18413294 TI - The removal of MO molecules from aqueous solution by the combination of ultrasound/adsorption/photocatalysis. AB - Methyl orange (MO) molecules had been removed from aqueous solution by nano TiO(2)/exfoliated graphite composites with the combination of ultrasound/adsorption/photocatalysis. The experimental results had showed that nano-TiO(2)/exfoliated graphite composites provided with the adsorptive capability of exfoliated graphite and the photocatalysis capability of nano TiO(2) synchronously. It had been proved that the removal ratios related to the adsorption and photocatalysis capability of the composites and were influenced by US and UV irradiation, the reaction times, the reaction temperatures, the initial pH values, the dosages and the initial MO solution concentrations. Moreover, it had been illuminated that the combination was very effective and useful on removing MO molecules from aqueous solution. PMID- 18413295 TI - Totally epicardial cardiac resynchronization therapy system implantation in patients with heart failure undergoing CABG. AB - BACKGROUND: Systolic dyssynchrony is present in a considerable number of patients with heart failure (HF) undergoing coronary artery bypass grafting (CABG). Surgical revascularization offers an optimal setting for totally epicardial cardiac resynchronization therapy (CRT) system implantation. AIM: To assess the efficacy of totally epicardial CRT implantation during CABG, in patients with HF. METHODS: Twenty three patients with HF and dyssynchrony underwent totally epicardial CRT system implantation during CABG. This randomised, single-blind, cross-over study compared clinical and echocardiographic parameters during two periods: 3 months of active CRT (CRT+) and 3 months of inactive CRT (CRT-) pacing. RESULTS: Twenty two patients underwent randomisation and completed both study periods. In the CRT+ group more patients improved by two NYHA classes (p=0.028), had a longer 6-minute walk test distance (p=0.047) and better quality of life (p=0.003) compared with the CRT- group. Echocardiography revealed an improved LV ejection fraction (p<0.001), smaller LV end-systolic volume (p=0.04), reduced mitral regurgitation (p=0.026) and improved LV synchrony in the CRT+ group compared with the CRT- group. CONCLUSION: CRT delivered by a totally epicardial system implanted during CABG is associated with additional improvement of clinical and echocardiographic parameters in patients with HF and systolic dyssynchrony. PMID- 18413296 TI - ABBA regulates plasma-membrane and actin dynamics to promote radial glia extension. AB - Radial glia play key roles in neuronal migration, axon guidance, and neurogenesis during development of the central nervous system. However, the molecular mechanisms regulating growth and morphology of these extended cells are unknown. We show that ABBA, a novel member of the IRSp53-MIM protein family, is enriched in different types of radial glia. ABBA binds ATP-actin monomers with high affinity and deforms PtdIns(4,5)P(2)-rich membranes in vitro through its WH2 and IM domains, respectively. In radial-glia-like C6-R cells, ABBA localises to the interface between the actin cytoskeleton and plasma membrane, and its depletion by RNAi led to defects in lamellipodial dynamics and process extension. Together, this study identifies ABBA as a novel regulator of actin and plasma membrane dynamics in radial glial cells, and provides evidence that membrane binding and deformation activity is critical for the cellular functions of IRSp53-MIM-ABBA family proteins. PMID- 18413298 TI - Clonal spread of highly beta-lactam-resistant Streptococcus pneumoniae isolates in Taiwan. AB - This study aimed to evaluate the antimicrobial susceptibility profiles of 364 Streptococcus pneumoniae isolates and studied the genotypes of S. pneumoniae with high level beta-lactam resistance in Taiwan. Clonal complexes related to Spain(23F)-1, Taiwan(19F)-14, and Taiwan(23F)-15 were responsible for the spread of isolates with high beta-lactam resistance. PMID- 18413297 TI - Thrombin hydrolysis of human osteopontin is dependent on thrombin anion-binding exosites. AB - The cytokine osteopontin (OPN) can be hydrolyzed by thrombin exposing a cryptic alpha(4)beta(1)/alpha(9)beta(1) integrin-binding motif (SVVYGLR), thereby acting as a potent cytokine for cells bearing these activated integrins. We show that purified milk OPN is a substrate for thrombin with a k(cat)/K(m) value of 1.14 x 10(5) m(-1) s(-1). Thrombin cleavage of OPN was inhibited by unsulfated hirugen (IC(50) = 1.2 +/- 0.2 microm), unfractionated heparin (IC(50) = 56.6 +/- 8.4 microg/ml) and low molecular weight (5 kDa) heparin (IC(50) = 31.0 +/- 7.9 microg/ml), indicating the involvement of both anion-binding exosite I (ABE-I) and anion-binding exosite II (ABE-II). Using a thrombin mutant library, we mapped residues important for recognition and cleavage of OPN within ABE-I and ABE-II. A peptide (OPN-(162-197)) was designed spanning the OPN thrombin cleavage site and a hirudin-like C-terminal tail domain. Thrombin cleaved OPN-(162-197) with a specificity constant of k(cat)/K(m) = 1.64 x 10(4) m(-1) s(-1). Representative ABE-I mutants (K65A, H66A, R68A, Y71A, and R73A) showed greatly impaired cleavage, whereas the ABE-II mutants were unaffected, suggesting that ABE-I interacts principally with the hirudin-like OPN domain C-terminal and contiguous to the thrombin cleavage site. Debye-Huckel slopes for milk OPN (-4.1 +/- 1.0) and OPN-(162-197) (-2.4 +/- 0.2) suggest that electrostatic interactions play an important role in thrombin recognition and cleavage of OPN. Thus, OPN is a bona fide substrate for thrombin, and generation of thrombin-cleaved OPN with enhanced pro-inflammatory properties provides another molecular link between coagulation and inflammation. PMID- 18413299 TI - Vitamin D and calcium dyshomoeostasis-associated heart failure. PMID- 18413300 TI - Cutaneous lesions and vitamin B12 deficiency: an often-forgotten link. PMID- 18413301 TI - Imaging breast augmentation and reconstruction. AB - Breast augmentation and breast reconstruction, either immediate or delayed, are increasingly common operations. All radiologists need to be able to recognize the normal appearances of the more commonly used implants on various imaging modalities, and breast radiologists in particular are facing new challenges when imaging the women involved. This article aims to review the normal and abnormal findings in women who have had breast augmentation and reconstruction, including implant insertion and reconstruction by autologous myocutaneous flaps. PMID- 18413302 TI - Fetal radiation dose from CT pulmonary angiography in late pregnancy: a phantom study. AB - Pulmonary embolism (PE) is the leading direct cause of maternal mortality in the UK. Accurate diagnosis is important but, even though CT pulmonary angiography (CTPA) is the recommended imaging modality for PE in the general population, there is limited guidance for pregnant patients. Knowledge of the radiation doses to both the mother and the fetus is therefore important in the justification of CTPA in this situation. Dose measurements were made on three helical CT scanners, with an anthropomorphic phantom representing the chest and abdomen in late gestation. Estimated fetal doses from CT scans of the maternal chest were in the range of 60-230 microGy. Fetal dose reduction strategies (mA modulation, shielding with a lead coat, and a 5 cm shorter scan length) were investigated. These reduced the fetal dose by 10%, 35% and 56%, respectively. Fetal doses from a scan projection radiograph (SPR) of the maternal chest were insignificant when compared with the dose from a CT scan. However, if the SPR was not stopped before the "fetus" was directly irradiated, the dose measured on one scanner was 20 microGy. PMID- 18413303 TI - Ethical recruitment of patients for PGS trial. PMID- 18413305 TI - The importance of the organization of partner knowledge in understanding perceptions of relationship quality and conflict resolution behavior in married couples. AB - This research investigated how the organization of partner knowledge was related to global relationship evaluations and specific interpersonal behaviors in a sample of 107 married couples. Spouses first completed several questionnaires and later participated in a 12-min videotaped conflict resolution task. As expected, wives in older relationships exhibited greater marital quality (self-reported and observed) when they had (a) an integrative knowledge structure and used many negative traits to describe their spouse or (b) a compartmentalized knowledge structure and used few negative traits to describe their spouse. Greater marital quality, however, was found for husbands when they were (a) in older relationships and possessed an integrative structure or (b) in newer relationships and possessed a compartmentalized structure. Exploratory analyses suggested that in some circumstances people may have greater marital quality when they share an organizational style with their spouse. PMID- 18413304 TI - Hypoxia is responsible for soluble vascular endothelial growth factor receptor-1 (VEGFR-1) but not for soluble endoglin induction in villous trophoblast. AB - BACKGROUND: Pre-eclampsia is a pregnancy disorder characterized by a maternal endothelial cell dysfunction associated with low levels of circulating placental growth factor (PlGF) and increased levels of total vascular endothelial growth factor (VEGF), soluble VEGF receptor-1 (sVEGFR-1), and soluble endoglin, a transforming growth factor beta1 and 3 coreceptor. Here, we tested the hypothesis that these altered levels of angiogenic cytokines and of the anti-angiogenic soluble forms of cytokine receptors could be the consequence of hypoxia. METHODS: Normal human umbilical vein endothelial cells, immortalized first trimester extravillous trophoblast cells (HTR8/SVneo) and first trimester placental villi explants (8-14 weeks) were used for culture under normoxia (20% O(2)) or hypoxia (1% O(2)). Culture media were collected for the measurement of cytokines by enzyme-linked immunosorbent assay. Total RNA was extracted for RT-PCR analysis. RESULTS: Under hypoxia, villous trophoblast expressed higher levels of VEGF, VEGFR-1, sVEGFR-1 and VEGFR-2 mRNAs (P < 0.001), and secreted more VEGF and sVEGFR-1 proteins (P < 0.05). In contrast, PlGF mRNA and protein were decreased in 1% O(2) (P < 0.001), whereas endoglin (Eng) was not modulated. Additionally, sVEGFR-1 directly abolished VEGF/PlGF-induced angiogenesis in the rat aortic ring assay. CONCLUSIONS: Our results support the hypotheses that, in pre-eclampsia, (i) overproduction of VEGF family factors by pre-eclamptic placenta is a consequence of induced hypoxia; (ii) overproduction of sVEGFR-1 by hypoxic villous trophoblast accounts for maternal free VEGF depletion; (iii) low circulating level of free PlGF is not only related to sVEGFR-1 overproduction, but also to hypoxia-induced mRNA down-regulation; (iv) Eng is not modulated by hypoxia. PMID- 18413306 TI - Is there benefit of cardiac slowing drugs in the treatment of hypertensive patients with elevated heart rate? PMID- 18413307 TI - Mild renal dysfunction associated with incident coronary artery disease in young males (from MELANY Study Investigators). PMID- 18413308 TI - Gene-load score of the renin-angiotensin-aldosterone system is associated with coronary heart disease in familial hypercholesterolaemia. AB - AIMS: Familial hypercholesterolaemia (FH) is characterized by premature coronary heart disease (CHD). However, the incidence of CHD varies considerably among FH patients. Genetic variation in the renin-angiotensin-aldosterone system (RAAS) and the adrenalin/noradrenalin system may be of importance in determining the CHD risk in FH, because of their involvement in CHD. We investigated the association between CHD risk and combined genetic variation in the RAAS and adrenalin/noradrenalin system. METHODS AND RESULTS: In 2190 FH patients, we genotyped six RAAS polymorphisms and five adrenalin/noradrenalin polymorphisms. For each patient, we calculated two gene-load scores by counting the number of risk genotypes within each pathway. Four of the six RAAS polymorphisms and none of the polymorphisms in the adrenalin/noradrenalin system were significantly associated with CHD (P < 0.05). The RAAS gene-load score was significantly associated with CHD (P(linear trend) < 0.001): in patients with a gene-load score of 5 or 6, the CHD risk was 2.3 times as high as in patients with a score of 0 or 1. The gene-load score of the adrenalin/noradrenalin system was not associated with CHD. CONCLUSION: Genetic variation in the RAAS contributes gene-dose dependently to CHD risk in patients with FH, whereas genetic variation in the adrenalin/noradrenalin system is not associated with CHD. PMID- 18413309 TI - Impact of diabetes on outcomes in patients with low and preserved ejection fraction heart failure: an analysis of the Candesartan in Heart failure: Assessment of Reduction in Mortality and morbidity (CHARM) programme. AB - AIMS: To determine whether the risk of adverse cardiovascular (CV) outcomes associated with diabetes differs in patients with low and preserved ejection fraction (EF) heart failure (HF). METHODS AND RESULTS: We analysed outcomes in the Candesartan in Heart failure-Assessment of Reduction in Mortality and morbidity (CHARM) programme which randomized 7599 patients with symptomatic HF and a broad range of EF. The prevalence of diabetes was 28.3% in patients with preserved EF (>40%) and 28.5% in those with low EF (15 years, and higher viral load, whereas death of a family member was associated with better retention. Among uninfected children, sites accruing low numbers of subjects, social worker responsible for retention, young age (1-2 years), and birth abnormalities were associated with higher risk of LTFU. Occurrences of certain stressful life events, such as a death in the family or financial instability, were associated with higher retention, but risk of LTFU increased when children started school or mothers began employment. Although participant characteristics are difficult to modify, the authors identified several potentially modifiable site practices that could be targeted to improve retention. PMID- 18413363 TI - Glutathione S-transferase Pi mediates proliferation of androgen-independent prostate cancer cells. AB - Prostate cancers generally acquire an androgen-independent growth capacity with progression, resulting in resistance to antiandrogen therapy. Therefore, identification of the genes regulated through this process may be important for understanding the mechanisms of prostate carcinogenesis. We here utilized androgen-dependent/independent transplantable tumors, newly established with the 'transgenic rat adenocarcinoma in prostate' (TRAP) model, to analyze their gene expression using microarrays. Among the overexpressed genes in androgen independent prostate cancers compared with the androgen-dependent tumors, glutathione S-transferase pi (GST-pi) was included. In line with this, human prostate cancer cell lines PC3 and DU145 (androgen independent) had higher expression of GST-pi compared with LNCaP (androgen dependent) as determined by semiquantitative reverse transcription-polymerase chain reaction analysis. To investigate the roles of GST-pi expression in androgen-independent human prostate cancers, GST-pi was knocked down by a small interfering RNA (siRNA), resulting in significant decrease of the proliferation rate in the androgen-independent PC3 cell line. In vivo, administration of GST-pi siRNA-atelocollagen complex decreased GST-pi protein expression, resulting in enhanced numbers of TdT mediated dUTP-biotin nick-end labering (TUNEL)-positive apoptotic cells. These findings suggest that GST-pi might play important roles in proliferation of androgen-independent human prostate cancer cells. PMID- 18413364 TI - Nrf2 enhances resistance of cancer cells to chemotherapeutic drugs, the dark side of Nrf2. AB - Drug resistance during chemotherapy is the major obstacle to the successful treatment of many cancers. Here, we report that inhibition of NF-E2-related factor 2 (Nrf2) may be a promising strategy to combat chemoresistance. Nrf2 is a critical transcription factor regulating a cellular protective response that defends cells against toxic insults from a broad spectrum of chemicals. Under normal conditions, the low constitutive amount of Nrf2 protein is maintained by the Kelch-like ECH-associated protein1 (Keap1)-mediated ubiquitination and proteasomal degradation system. Upon activation, this Keap1-dependent Nrf2 degradation mechanism is quickly inactivated, resulting in accumulation and activation of the antioxidant response element (ARE)-dependent cytoprotective genes. Since its discovery, Nrf2 has been viewed as a 'good' transcription factor that protects us from many diseases. In this study, we demonstrate the dark side of Nrf2: stable overexpression of Nrf2 resulted in enhanced resistance of cancer cells to chemotherapeutic agents including cisplatin, doxorubicin and etoposide. Inversely, downregulation of the Nrf2-dependent response by overexpression of Keap1 or transient transfection of Nrf2-small interfering RNA (siRNA) rendered cancer cells more susceptible to these drugs. Upregulation of Nrf2 by the small chemical tert-butylhydroquinone (tBHQ) also enhanced the resistance of cancer cells, indicating the feasibility of using small chemical inhibitors of Nrf2 as adjuvants to chemotherapy to increase the efficacy of chemotherapeutic agents. Furthermore, we provide evidence that the strategy of using Nrf2 inhibitors to increase efficacy of chemotherapeutic agents is not limited to certain cancer types or anticancer drugs and thus can be applied during the course of chemotherapy to treat many cancer types. PMID- 18413365 TI - Bioenergetic differences selectively sensitize tumorigenic liver progenitor cells to a new gold(I) compound. AB - A hallmark of cancer cells is their ability to evade apoptosis and mitochondria play a critical role in this process. Delineating mitochondrial differences between normal and cancer cells has proven challenging due to the lack of matched cell lines. Here, we compare two matched liver progenitor cell (LPC) lines, one non-tumorigenic [p53-immortalized liver (PIL) 4] and the other tumorigenic (PIL2). Analysis of these cell lines and a p53 wild-type non-tumorigenic cell line [bipotential murine oval liver (BMOL)] revealed an increase in expression of genes encoding the antiapoptotic proteins cellular inhibitor of apoptosis protein (cIAP) 1 and yes associate protein in the PIL2 cells, which resulted in an increase in the protein encoded by these genes. PIL2 cells have higher mitochondrial membrane potential (Deltapsi(m)) compared with PIL4 and BMOL and had greater levels of reactive oxygen species, despite the fact that the mitochondrial antioxidant enzyme, manganese superoxide disumutase, was elevated at transcript and protein levels. Taken together, these results may account for the observed resistance of PIL2 cells to apoptotic stimuli compared with PIL4. We tested a new gold compound to show that hyperpolarized Deltapsi(m) led to its increased accumulation in mitochondria of PIL2 cells. This compound selectively induces apoptosis in PIL2 cells but not in PIL4 or BMOL. The gold compound depolarized the Deltapsi(m), depleted the adenosine triphosphate pool and activated caspase-3 and caspase-9, suggesting that apoptosis was mediated via mitochondria. This investigation shows that the non-tumorigenic and tumorigenic LPCs are useful models to delineate the role of mitochondrial dysfunction in tumorigenesis and for the future development of mitochondria-targeted chemotherapeutics that selectively target tumor cells. PMID- 18413366 TI - Glypican-3-mediated oncogenesis involves the Insulin-like growth factor-signaling pathway. AB - Glypican-3 (gpc3) is the gene responsible for Simpson-Golabi-Behmel overgrowth syndrome. Previously, we have shown that GPC3 is overexpressed in hepatocellular carcinoma (HCC). In this study, we demonstrated the mechanisms for GPC3-mediated oncogenesis. Firstly, GPC3 overexpression in NIH3T3 cells gave to cancer cell phenotypes including growing in serum-free medium and forming colonies in soft agar, or on the other way, GPC3 knockdown in HuH-7 cells decreased oncogenecity. We further demonstrated that GPC3 bound specifically through its N-terminal proline-rich region to both Insulin-like growth factor (IGF)-II and IGF-1R. GPC3 stimulated the phosphorylation of IGF-1R and the downstream signaling molecule extracellular signal-regulated kinase (ERK) in an IGF-II-dependent way. Also, GPC3 knockdown in HCC cells decreased the phosphorylation of both IGF-1R and ERK. Therefore, GPC3 confers oncogenecity through the interaction between IGF-II and its receptor, and the subsequent activation of the IGF-signaling pathway. This data are novel to the current understanding of the role of GPC3 in HCC and will be important in future developments of cancer therapy. PMID- 18413367 TI - A novel role of thrombospondin-1 in cervical carcinogenesis: inhibit stroma reaction by inhibiting activated fibroblasts from invading cancer. AB - Thrombospondin (TSP)-1, a potent angiogenesis inhibitor, has been shown to exert different biological functions on various cell types. Here, we investigate the role of TSP-1 in tumor-stroma reaction, which is mainly characterized by fibroblast activation to create a permissive microenvironment for tumor progression. Immunohistochemistry examinations in the human surgical specimens have shown that a downregulation of TSP-1 during the progression of cervical carcinogenesis was accompanied by an emergence in the upregulation of stroma markers, alpha-smooth muscle actin (alpha-SMA) and desmin. Transfection of SiHa cervical cancer cells with a plasmid expressing the TSP-1 protein exhibited antiangiogenic activity in vitro and resulted in reduced tumor growth in severe combined immunodeficiency (SCID) mice, which was accompanied by a decrease in tumor vascularization and lower expressions of alpha-SMA and desmin than those in the vector controls. Transfection with TSP-1 and purified TSP-1 added to NIH3T3 cells did not alter the protein levels of alpha-SMA and desmin but significantly inhibited matrix metalloprotease-2 activity. Transforming growth factor-beta (TGF beta), a major factor in the activation of fibroblasts, increased alpha-SMA and desmin expression and the ability of cell migration and invasion in NIH3T3 cells. The increased migration ability and the invasive ability into tumor cluster of TGF-beta-treated NIH3T3 cells were dose dependently inhibited by TSP-1. In contrast, ectopic TSP-1 expression in SiHa cells has little effect on the invasive ability of the NIH3T3 cells. Together, our findings demonstrate a novel role of TSP-1 to inhibit tumor-stroma reaction that could be attributed to the blockage of activated fibroblasts from invading cancer cells. PMID- 18413368 TI - Meta-analysis of vascular endothelial growth factor variations in amyotrophic lateral sclerosis: increased susceptibility in male carriers of the -2578AA genotype. AB - BACKGROUND: Targeted delivery of the angiogenic factor, vascular endothelial growth factor (VEGF), to motor neurons prolongs survival in rodent models of amyotrophic lateral sclerosis (ALS), while mice expressing reduced VEGF concentrations develop motor neuron degeneration reminiscent of ALS, raising the question whether VEGF contributes to the pathogenesis of ALS. An initial association study reported that VEGF haplotypes conferred increased susceptibility to ALS in humans, but later studies challenged this initial finding. METHODS AND FINDINGS: A meta-analysis was undertaken to critically reappraise whether any of the three common VEGF gene variations (-2578C/A, 1154G/A and -634G/C) increase the risk of ALS. Over 7000 subjects from eight European and three American populations were included in the analysis. Pooled odds ratios were calculated using fixed-effects and random-effects models, and four potential sources of heterogeneity (location of disease onset, gender, age at disease onset and disease duration) were assessed. After correction, none of the genotypes or haplotypes was significantly associated with ALS. Subgroup analysis by gender revealed, however, that the -2578AA genotype, which lowers VEGF expression, increased the risk of ALS in males (OR = 1.46 males vs females; 95% CI = 1.19 to 1.80; p = 7.8 10E-5), even after correction for publication bias and multiple testing. CONCLUSIONS: This meta-analysis does not support the original conclusion that VEGF haplotypes increase the risk of ALS in humans, but the significant association of the low-VEGF -2578AA genotype with increased susceptibility to ALS in males reappraises the link between reduced VEGF concentrations and ALS, as originally revealed by the fortuitous mouse genetic studies. PMID- 18413369 TI - Expansion in size of a terminal deletion: a paradigm shift for parental follow-up studies. AB - BACKGROUND: Parental studies are often necessary subsequent to the identification of a chromosome abnormality. The recommended studies are based on assumptions about how chromosome rearrangements occur. One such assumption is that deletion size is stable through generations. RESULTS: We have identified a family where a small subtelomeric deletion in a phenotypically and cytogenetically normal mother expanded nearly 10-fold into a clinically consequential and cytogenetically visible deletion in her affected daughter. CONCLUSION: Traditional parental follow-up studies would have not identified this expansion, but would have rather classified the deletion in the daughter as either de novo or benign. Only by sizing the deletion by array comparative genomic hybridisation in both the mother and the daughter was the expansion recognised. Previous assumptions about chromosome behaviour suggest that this phenomenon may have been easily missed in other cases of chromosomal deletions. Therefore, this case illustrates the need for more comprehensive analyses of parental chromosome structure when characterising an abnormality in a child. PMID- 18413370 TI - X-linked ichthyosis (steroid sulfatase deficiency) is associated with increased risk of attention deficit hyperactivity disorder, autism and social communication deficits. AB - BACKGROUND: X-linked ichthyosis (XLI) (steroid sulfatase deficiency) is caused by deletions or point mutations of the steroid sulfatase (STS) gene on chromosome Xp22.32. Deletions of this region can be associated with cognitive behavioural difficulties including autism. Animal work suggests the STS gene may be involved in attentional processes. We have therefore undertaken a systematic study of autism and attention deficit hyperactivity disorder (ADHD) in boys with XLI. METHODS: Cases of XLI were recruited from families originally ascertained when pregnancies with STS deficiency were identified through a routine maternal screening programme. Boys with XLI were assessed for ADHD and autism using standardised questionnaires and interviews. Deletions of the STS gene were identified and characterised by analysis of genomic DNA and/or fluorescent in situ hybridisation. RESULTS: 25 boys with XLI were assessed for autism and ADHD. 40% fulfilled DSM-IV criteria for a diagnosis of ADHD, 80% of which were inattentive subtype. ADHD diagnoses were present in those with both deletions and presumed point mutations of STS. Additionally, five boys, from three unrelated families, fulfilled criteria for an autistic spectrum disorder or related language/communication difficulty, and all had an unusually large deletion of the STS gene with loss of the neuroligin 4 (NLGN4) gene. None of the boys with the typical deletion or presumed point mutations of STS demonstrated autistic difficulties. CONCLUSIONS: STS deficiency may be a risk factor for ADHD with predominantly inattentive symptoms. Boys with XLI and large deletions encompassing STS and NLGN4 are at increased risk of developing autism and related disorders. PMID- 18413371 TI - The fragile X prevalence paradox. PMID- 18413372 TI - Screening for familial ovarian cancer: poor survival of BRCA1/2 related cancers. AB - AIM: To assess the effectiveness of annual ovarian cancer screening (transvaginal ultrasound and serum CA125 estimation) in reducing mortality from ovarian cancer in women at increased genetic risk. PATIENTS AND METHODS: A cohort of 3532 women at increased risk of ovarian cancer was screened at five European centres between January 1991 and March 2007. Survival from diagnosis of ovarian cancer was calculated using Kaplan-Meier analysis and compared for proven BRCA1/2 carriers with non-carriers and whether the cancer was detected at prevalence or post prevalent scan. Screening was performed by annual transvaginal ultrasound and serum CA125 measurement. RESULTS: 64 epithelial ovarian malignancies (59 invasive and 5 borderline), developed in the cohort. 26 tumours were detected at prevalent round; there were 27 incident detected cancers and 11 interval. 65% of cancers were stage 3 or 4, however, stage and survival were little different for prevalent versus post-prevalent cancers. Five year and 10 year survival in 49 BRCA1/2 mutation carriers was 58.6% (95% CI 50.9% to 66.3%) and 36% (95% CI 27% to 45%), which was significantly worse than for 15 non-BRCA carriers (91.8%, 95% CI 84% to 99.6%, both 5 and 10 year survival p = 0.015). However, when borderline tumours were excluded, the difference in survival between carriers and non carriers was no longer significant. CONCLUSION: Annual surveillance, by transvaginal ultrasound scanning and serum CA125 measurement, in women at increased familial risk of ovarian cancer is ineffective in detecting tumours at a sufficiently early stage to influence substantially survival in BRCA1/2 carriers. PMID- 18413373 TI - Detection of known and novel genomic rearrangements by array based comparative genomic hybridisation: deletion of ZNF533 and duplication of CHARGE syndrome genes. AB - BACKGROUND: Mental retardation can be caused by copy number variations (deletions, insertions, duplications), ranging in size from 1 kb to several megabases. Array based comparative genomic hybridisation (array-CGH) allows detection of an increasing number of genomic alterations. METHODS: A series of 46 patients with mental retardation and congenital abnormalities (previously screened for subtelomeric rearrangements) were evaluated for cryptic chromosomal imbalances by array-CGH. This array contains 6465 large-insert BAC/PAC clones, representing sequences uniformly distributed throughout the human genome. The results were confirmed by alternative techniques. RESULTS: Four pathogenic rearrangements were detected: two of them were novel, a deletion at 2q31.2 and a duplication at 8q12 band; the other two have been previously reported--a duplication of the Williams-Beuren region and a deletion of 3q29. By adding the subtelomeric alterations previously identified, a total rate of 18% of pathogenic rearrangements was found in the series. CONCLUSION: Based on our results, ZNF533 is the only gene contained in the overlapping region with other deletions at 2q31.2, and it is most probably the fourth zinc-finger gene implied in mental retardation. On the other hand, we propose that the CHD7 gene, associated with CHARGE syndrome by haploinsufficiency, causes a different phenotype by gain-of dosage. PMID- 18413374 TI - Predicting the likelihood of carrying a BRCA1 or BRCA2 mutation: validation of BOADICEA, BRCAPRO, IBIS, Myriad and the Manchester scoring system using data from UK genetics clinics. AB - OBJECTIVES: Genetic testing for the breast and ovarian cancer susceptibility genes BRCA1 and BRCA2 has important implications for the clinical management of people found to carry a mutation. However, genetic testing is expensive and may be associated with adverse psychosocial effects. To provide a cost-efficient and clinically appropriate genetic counselling service, genetic testing should be targeted at those individuals most likely to carry pathogenic mutations. Several algorithms that predict the likelihood of carrying a BRCA1 or a BRCA2 mutation are currently used in clinical practice to identify such individuals. DESIGN: We evaluated the performance of the carrier prediction algorithms BOADICEA, BRCAPRO, IBIS, the Manchester scoring system and Myriad tables, using 1934 families seen in cancer genetics clinics in the UK in whom an index patient had been screened for BRCA1 and/or BRCA2 mutations. The models were evaluated for calibration, discrimination and accuracy of the predictions. RESULTS: Of the five algorithms, only BOADICEA predicted the overall observed number of mutations detected accurately (ie, was well calibrated). BOADICEA also provided the best discrimination, being significantly better (p<0.05) than all models except BRCAPRO (area under the receiver operating characteristic curve statistics: BOADICEA = 0.77, BRCAPRO = 0.76, IBIS = 0.74, Manchester = 0.75, Myriad = 0.72). All models underpredicted the number of BRCA1 and BRCA2 mutations in the low estimated risk category. CONCLUSIONS: Carrier prediction algorithms provide a rational basis for counselling individuals likely to carry BRCA1 or BRCA2 mutations. Their widespread use would improve equity of access and the cost effectiveness of genetic testing. PMID- 18413375 TI - Diagnostic accuracy of 11C-methionine PET for differentiation of recurrent brain tumors from radiation necrosis after radiotherapy. AB - We evaluated the diagnostic accuracy of PET with l-methyl-(11)C-methionine ((11)C MET) for the differentiation of recurrent brain tumors from radiation necrosis. METHODS: Seventy-seven patients who had been previously treated with radiotherapy after primary treatment for metastatic brain tumor (n=51) or glioma (n=26) were studied to clarify the diagnostic performance of (11)C-MET PET in differentiating between recurrent brain tumors and radiation necrosis. A total of 88 PET scans with (11)C-MET were obtained; sometimes more than one scan was obtained when there was an indication of recurrent brain tumor or radiation necrosis. A definitive diagnosis was made on the basis of pathologic examination for recurrent brain tumors and on the basis of pathologic examination or clinical course for radiation necrosis. Several indices characterizing the lesions were determined; these included mean and maximum standardized uptake values (SUV(mean) and SUV(max), respectively) and the ratios of lesion uptake to contralateral normal frontal-lobe gray matter uptake corresponding to the SUV(mean) and the SUV(max) (L/N(mean) and L/N(max), respectively). Receiver-operating characteristic (ROC) curve analysis was used to determine the optimal index of (11)C-MET PET and cutoff values for the differential diagnosis of tumor recurrence and radiation necrosis. RESULTS: The values of each index of (11)C-MET PET tended to be higher for tumor recurrence than for radiation necrosis. There were significant differences between tumor recurrence and radiation necrosis in all of the indices except for the L/N(max) for glioma. ROC analysis indicated that the L/N(mean) was the most informative index for differentiating between tumor recurrence and radiation necrosis. An L/N(mean) of greater than 1.41 provided the best sensitivity and specificity for metastatic brain tumor (79% and 75%, respectively), and an L/N(mean) of greater than 1.58 provided the best sensitivity and specificity for glioma (75% and 75%, respectively). CONCLUSION: (11)C-MET PET can provide quantitative values to aid in the differentiation of tumor recurrence from radiation necrosis, although these values do not appear to be absolute indicators. Quantitative analysis of (11)C-MET PET data may be helpful in managing irradiated brain tumors. PMID- 18413376 TI - Myeloablative 131I-tositumomab radioimmunotherapy in treating non-Hodgkin's lymphoma: comparison of dosimetry based on whole-body retention and dose to critical organ receiving the highest dose. AB - Myeloablative radioimmunotherapy using (131)I-tositumomab (anti-CD20) monoclonal antibodies is an effective therapy for B-cell non-Hodgkin's lymphoma. The amount of radioactivity for radioimmunotherapy may be determined by several methods, including those based on whole-body retention and on dose to a limiting normal organ. The goal of each approach is to deliver maximal myeloablative amounts of radioactivity within the tolerance of critical normal organs. METHODS: Records of 100 consecutive patients who underwent biodistribution and dosimetry evaluation after tracer infusion of (131)I-tositumomab before radioimmunotherapy were reviewed. We assessed organ and tissue activities over time by serial gamma camera imaging to calculate radiation-absorbed doses. Organ volumes were determined from CT scans for organ-specific dosimetry. These dose estimates helped us to determine therapy on the basis of projected dose to the critical normal organ receiving a maximum tolerable radiation dose. We compared organ specific dosimetry for treatment planning with the whole-body dose-assessment method by retrospectively analyzing the differences in projected organ-absorbed doses and their ratios. RESULTS: Mean organ doses per unit of administered activity (mGy/MBq) estimated by both methods were 0.33 for liver and 0.33 for lungs by the whole-body method and 1.52 for liver and 1.74 for lungs by the organ specific method (P=0.0001). The median differences between methods were 0.92 mGy/MBq (range, 0.36-2.2 mGy/MBq) for lungs, 0.82 mGy/MBq (range, 0.28-1.67 mGy/MBq) for liver, and -0.01 mGy/MBq (range, -0.18-0.16 mGy/MBq) for whole body. The median ratios of the treatment activities based on limiting normal-organ dose were 5.12 (range, 2.33-10.01) for lungs, 4.14 (range, 2.16-6.67) for liver, and 0.94 (range, 0.79-1.22) for whole body. We found substantial differences between the dose estimated by the 2 methods for liver and lungs (P=0.0001). CONCLUSION: Dosimetry based on whole-body retention will underestimate the organ doses, and a preferable approach is to evaluate organ-specific doses by accounting for actual radionuclide biodistribution. Myeloablative treatments based on the latter approach allow administration of the maximum amount of radioactivity while minimizing toxicity. PMID- 18413378 TI - Recombinant human TSH-assisted radioactive iodine remnant ablation achieves short term clinical recurrence rates similar to those of traditional thyroid hormone withdrawal. AB - Recent studies have confirmed that radioactive iodine therapy after recombinant human TSH (rhTSH) stimulation effectively ablates the normal thyroid remnant. However, no published study has determined the effectiveness of rhTSH preparations on the important endpoint of disease recurrence. METHODS: Disease recurrence was retrospectively assessed a median of 2.5 y after radioiodine remnant ablation (RRA) in 394 consecutive thyroid cancer patients (93% papillary, 71% female, 47+/-15 y old [mean +/- SD], median (131)I dose of 3,996 MBq [108 mCi]). RESULTS: Similar rates of clinically evident disease recurrence (4% rhTSH vs. 7% thyroid hormone withdrawal [THW], P=not statistically significant) and residual thyroid bed uptake without other evidence of persistent disease (4% rhTSH vs. 7% THW, P=not statistically significant) were seen in the 320 patients undergoing rhTSH-assisted RRA and the 74 patients prepared for RRA by THW. When the definition of no clinical evidence of disease included a suppressed thyroglobulin level of less than 1 ng/mL and a stimulated thyroglobulin level of less than 2 ng/mL, rhTSH-assisted RRA was associated with significantly higher rates of no clinical evidence of disease (74% rhTSH vs. 55% THW, P=0.02) and significantly lower rates of persistent disease (19% rhTSH vs. 32% THW, P=0.02) than was RRA after THW. Patients selected for rhTSH-assisted RRA were older (48+/ 15 vs. 44+/-15 y, P=0.03) and received a slightly higher administered activity of (131)I (median, 4,033 MBq [109 mCi] vs. 3,811 MBq [103 mCi], P=0.01) but did not differ with respect to sex, histology, disease stage, or mean time to recurrence (19+/-9 mo for rhTSH vs. 20+/-16 mo for THW). CONCLUSION: rhTSH-assisted RRA is associated with rates of clinically evident disease recurrence and persistent uptake in the thyroid bed that are similar to those for traditional THW. PMID- 18413379 TI - Preparation of a promising angiogenesis PET imaging agent: 68Ga-labeled c(RGDyK) isothiocyanatobenzyl-1,4,7-triazacyclononane-1,4,7-triacetic acid and feasibility studies in mice. AB - Arg-Gly-Asp (RGD) derivatives have been labeled with various radioisotopes for the imaging of angiogenesis in ischemic tissue, in which alpha(v)beta(3) integrin plays an important role. In this study, cyclic Arg-Gly-Asp-D-Tyr-Lys [c(RGDyK)] was conjugated with 2-(p-isothiocyanatobenzyl)-1,4,7-triazacyclononane-1,4,7 triacetic acid (SCN-Bz-NOTA) and then labeled with (68)Ga. The labeled RGD so produced was subjected to an in vitro binding assay and in vivo biodistribution and PET studies. METHODS: A mixture of SCN-Bz-NOTA (660 nmol) and c(RGDyK) (600 nmol) in 0.1 M sodium carbonate buffer (pH 9.5) was allowed to react for 20 h at room temperature in the dark for thiourea bond formation. The conjugate obtained was purified by semipreparative high-performance liquid chromatography (HPLC). The purified c(RGDyK)-SCN-Bz-NOTA (NOTA-RGD) was then labeled with (68)Ga from a (68)Ge/(68)Ga generator and purified by semipreparative HPLC. A competitive binding assay for c(RGDyK) and NOTA-RGD was performed with (125)I-c(RGDyK) as a radioligand and alpha(v)beta(3) integrin-coated plates as a solid phase. (68)Ga NOTA-RGD (0.222 MBq/100 microL) was injected, through a tail vein, into mice with hind limb ischemia and into mice bearing human colon cancer SNU-C4 xenografts. Biodistribution and imaging studies were performed at 1 and 2 h after injection. RESULTS: The labeling of NOTA-RGD with (68)Ga was straightforward. The K(i) values of c(RGDyK) and NOTA-RGD were 1.3 and 1.9 nM, respectively. In the biodistribution study, the mean +/- SD uptake of (68)Ga-NOTA-RGD by ischemic muscles was 1.6+/-0.2 percentage injected dose per gram (%ID/g); this uptake was significantly blocked by cold c(RGDyK) to 0.6+/-0.3 %ID/g (P<0.01). Tumor uptake was 5.1+/-1.0 %ID/g, and the tumor-to-blood ratio was 10.3+/-4.8. Small-animal PET revealed rapid excretion through the urine and high levels of tumor and kidney uptake. CONCLUSION: Stable (68)Ga-NOTA-RGD was obtained in a straightforward manner at a high yield and showed a high affinity for alpha(v)beta(3) integrin, specific uptake by angiogenic muscles, a high level of uptake by tumors, and rapid renal excretion. (68)Ga-NOTA-RGD was found to be a promising radioligand for the imaging of angiogenesis. PMID- 18413380 TI - Radionuclide evaluation of the lower gastrointestinal tract. AB - This review outlines the technical aspects and diagnostic performance parameters of nuclear medicine procedures used on patients with disorders of the lower gastrointestinal tract, with the exclusion of techniques using tumor-seeking radiopharmaceuticals. Chronic disorders of the lower gastrointestinal tract often reduce the quality of life because of discomfort from constipation or diarrhea. Five classes of radionuclide procedures are used to characterize these disorders: transit scintigraphy, searches for ectopic gastric mucosa in Meckel's diverticulum, scintigraphy of active inflammatory bowel disease, scintigraphic defecography, and scintigraphy to detect sites of gastrointestinal bleeding. Protocols for these procedures and their relative merit in patient management are discussed, with special emphasis on their potential for semiquantitative assessment of the pathophysiologic parameter investigated. Quantitation is particularly relevant for prognostic purposes and for monitoring the efficacy of therapy. PMID- 18413381 TI - New 133Xe gas trapping index for quantifying severe emphysema before partial lung volume reduction. AB - Lung volume reduction (LVR) is an effective therapy for end-stage emphysema. Preliminary and postprocedure imaging is usually limited to CT for anatomic delineation of the location and severity of the most acutely affected lung zones. The purpose of this study was to investigate the potential of using a new quantitative gas trapping index (GTI) derived from a (133)Xe ventilation scan to assess the severity of emphysema. METHODS: Using the equilibration and washout phases of a (133)Xe ventilation study, a GTI was compared with visual National Emphysema Treatment Trial (NETT) CT scoring, semiautomated CT densitometry, and (99m)Tc perfusion scintigraphy in 28 patients being evaluated for LVR. The GTI was calculated as the percentage of (133)Xe gas retention in a 3-min washout image compared with the peak equilibrium image for 6 lung zones. RESULTS: The GTI correlated best with the percentage of perfusion (-0.39, P<0.0001) and the CT density scoring with the percentage of severe emphysema (0.36, P<0.0001). There was less correlation with visual NETT CT scores (0.25, P<0.001). CONCLUSION: This GTI, based on widely available (133)Xe imaging, shows good correlation with other quantitative measures of emphysema that are anatomically based. Because this GTI provides a more functional assessment of gas trapping and airway disease, these results suggest that additional study is warranted to investigate its use as a functional measure of emphysema before and after LVR. PMID- 18413382 TI - Molecular and functional MRI of the tumor microenvironment. PMID- 18413384 TI - Reducing radiation dose in rest-stress cardiac PET/CT by single poststress cine CT for attenuation correction: quantitative validation. AB - Cardiac PET/CT is optimized by cine CT with dedicated shift software for manual correction of attenuation-emission misregistration. Separate rest and stress CT scans incur greater radiation dose to patients than does standard helical PET/CT or "pure" PET using rotating rod attenuation sources. To reduce radiation dose, we tested quantitative accuracy of using a single poststress cine CT attenuation scan for reconstructing rest perfusion images to eliminate resting CT attenuation scans. METHODS: A total of 250 consecutive patients underwent diagnostic rest dipyridamole myocardial perfusion PET/CT with (82)Rb and a 16-slice PET/CT scanner using averaged cine CT attenuation data during breathing at rest and stress. After correcting for any attenuation-emission misregistration, we quantitatively compared resting perfusion images reconstructed using rest cine CT attenuation data with the same resting emission data reconstructed with poststress cine CT attenuation data. Automated software quantifying average regional quadrant activity, severity, size, and combined size and severity of perfusion defects was used for this comparison. RESULTS: Resting perfusion images reconstructed using rest cine CT attenuation data were quantitatively comparable to resting images reconstructed with poststress cine CT attenuation data with no clinically significant differences. Twenty-five (10%) of 250 cases required shifting of stress cine CT attenuation data to achieve optimal attenuation emission coregistration with resting perfusion data. Eliminating rest CT attenuation scans reduced CT radiation dose by 50% below rest-plus-stress cine CT protocols. CONCLUSION: Resting perfusion images reconstructed using poststress cine CT attenuation data are quantitatively comparable to resting images reconstructed with resting cine CT attenuation data. Eliminating the rest CT scan reduces CT radiation dose by 50%. PMID- 18413385 TI - Long-term precision of 18F-fluoride PET skeletal kinetic studies in the assessment of bone metabolism. AB - (18)F-Fluoride PET allows noninvasive evaluation of regional bone metabolism and has the potential to become a useful tool for assessing patients with metabolic bone disease and evaluating novel drugs being developed for these diseases. The main PET parameter of interest, termed K(i), reflects regional bone metabolism. The aim of this study was to compare the long-term precision of (18)F-fluoride PET with that of biochemical markers of bone turnover assessed over 6 mo. METHODS: Sixteen postmenopausal women with osteoporosis or significant osteopenia and a mean age of 64 y underwent (18)F-fluoride PET of the lumbar spine and measurements of biochemical markers of bone formation (bone-specific alkaline phosphatase and osteocalcin) and bone resorption (urinary deoxypyridinoline) at baseline and 6 mo later. Four different methods for analyzing the (18)F-fluoride PET data were compared: a 4k 3-compartmental model using nonlinear regression analysis (K(i-4k)), a 3k 3-compartmental model using nonlinear regression analysis (K(i-3k)), Patlak analysis (K(i-PAT)), and standardized uptake values. RESULTS: With the exception of a small but significant decrease in K(i-3k) at 6 mo, there were no significant differences between the baseline and 6-mo values for the PET parameters or biochemical markers. The long-term precision, expressed as the coefficient of variation (with 95% confidence interval in parentheses), was 12.2% (9%-19%), 13.8% (10%-22%), 14.4% (11%-22%), and 26.6% (19%-40%) for K(i 3k), K(i-PAT), mean standardized uptake value, and K(i-4k), respectively. For comparison, the precision of the biochemical markers was 10% (7%-15%), 18% (13% 27%), and 14% (10%-21%) for bone-specific alkaline phosphatase, osteocalcin, and urinary deoxypyridinoline, respectively. Intraclass correlation between the baseline and 6-mo values ranged from 0.44 for K(i-4k) to 0.85 for K(i-3k). No significant correlation was found between the repeated mean standardized uptake value measurements. CONCLUSION: The precision and intraclass correlation observed for K(i-3k) and K(i-PAT) was equivalent to that observed for biochemical markers. This study provided initial data on the long-term precision of (18)F-fluoride PET measured at the lumbar spine, which will aid in the accurate interpretation of changes in regional bone metabolism in response to treatment. PMID- 18413386 TI - Diastolic filling parameters derived from myocardial perfusion imaging can predict left ventricular end-diastolic pressure at subsequent cardiac catheterization. AB - Morbidity and mortality increase when diastolic dysfunction accompanies coronary artery disease (CAD). An elevated stress (201)Tl lung-to-heart ratio (LHR) is a traditional marker of elevated left ventricular end-diastolic pressure (LVEDP), which adds prognostic value in CAD. Since the introduction of (99m)Tc-labeled agents, this valuable marker has been lost. Hence, there is only a limited ability to assess diastolic dysfunction by myocardial perfusion imaging (MPI). METHODS: Fifty-two consecutive patients with an ejection fraction of >or=45% underwent MPI and cardiac catheterization within 15 d. Peak filling rate (PFR), time to PFR (TPFR), and filling rate during the first third of diastole (1/3FR) were obtained from MPI with SPECT software. Resting (201)Tl LHR was calculated manually, and LVEDP was obtained at catheterization. RESULTS: PFR, TPFR, and 1/3FR correlated significantly with LVEDP (r= -0.53, 0.45, and -0.45, respectively; P=0.00005, 0.0009, and 0.0009, respectively), whereas resting (201)Tl LHR did not (r=0.10, P=0.49). Receiver-operating-characteristic curve analysis of PFR, TPFR, and 1/3FR for detecting LVEDPs of >or=18 mm Hg showed areas under the curve of 0.83, 0.75, and 0.80, respectively. The combination of PFR and 1/3FR showed a negative predictive value of 84%, a positive predictive value of 86%, and a specificity of 94%. CONCLUSION: Diastolic filling variables obtained with the SPECT software showed a significant correlation with LVEDP. PFR, TPFR, and 1/3FR were superior to resting (201)Tl LHR and showed good sensitivity, specificity, and predictive power for detecting LVEDPs of >or=18 mm Hg. Hence, combining data on the presence of perfusion defects with data on diastolic impairments can be achieved by adding these variables to MPI results. PMID- 18413387 TI - Visualization of brown adipose tissue with 99mTc-methoxyisobutylisonitrile on SPECT/CT. AB - Brown adipose tissue (BAT) is retained into adulthood in some patients. It has been imaged using several radiopharmaceuticals, including (18)F-FDG. Using SPECT/CT, we assessed whether and how frequently uptake of (99m)Tc methoxyisobutylisonitrile ((99m)Tc-MIBI) was present in expected locations of BAT. METHODS: A total of 205 SPECT/CT scans using (99m)Tc-MIBI for parathyroid imaging were reviewed for the presence of (99m)Tc-MIBI uptake in expected locations of BAT. RESULTS: We detected (99m)Tc-MIBI uptake in BAT in 13 of 205 patients (6.3%). When BAT was visualized, it was detected on both early and delayed scans. The patients with (99m)Tc-MIBI uptake in BAT were younger than the patients with no (99m)Tc-MIBI uptake (P=0.044). CONCLUSION: Uptake of (99m)Tc MIBI in BAT is relatively common in this adult patient population and should not be confused with (99m)Tc-MIBI-avid-tumors. SPECT/CT can be useful for defining such tracer uptake as a normal physiologic variant. PMID- 18413388 TI - A new acycloguanosine-specific supermutant of herpes simplex virus type 1 thymidine kinase suitable for PET imaging and suicide gene therapy for potential use in patients treated with pyrimidine-based cytotoxic drugs. AB - The herpes simplex virus type 1 thymidine kinase (HSV1-tk) gene is widely used as a suicide gene in combination with ganciclovir (GCV) and as a nuclear imaging reporter gene with an appropriate reporter probe. Wild-type HSV1-tk recognizes a variety of pyrimidine and acycloguanosine nucleoside analogs, including clinically used antiviral drugs. PET of HSV1-tk reporter gene expression will be compromised in patients receiving nucleoside-based antiviral treatment. With the use of an acycloguanosine-specific mutant of the enzyme, PET of HSV1-tk reporter gene expression can be successfully performed with acycloguanosine-based radiotracers without interference from pyrimidine-based antiviral drugs. METHODS: The levels of expression of wild-type HSV1-tk and HSV1-A167Ytk, HSV1-sr39tk, and HSV1-A167Ysr39tk mutants fused with green fluorescent protein (GFP) and transduced into U87 cells were normalized to the mean fluorescence of GFP measured by fluorescence-activated cell sorting. The levels of enzymatic activities of wild-type HSV1-tk and its mutants were compared by 2-h in vitro radiotracer uptake assays with (3)H-2'-fluoro-2'-deoxy-1-beta-d-arabinofuranosyl 5-ethyluracil ((3)H-FEAU), (3)H-pencyclovir ((3)H-PCV), and (3)H-GCV and by drug sensitivity assays. PET with (18)F-FEAU and (18)F-9-[4-fluoro-3 (hydroxymethyl)butyl]guanine ((18)F-FHBG) was performed in mice with established subcutaneous tumors, expressing wild-type HSV1-tk and its mutants, followed by tissue sampling. RESULTS: FEAU accumulation was not detected in HSV1-A167Ysr39tk expressing cells and xenografts. Lack of conversion of pyrimidine derivatives by the HSV1-A167Ysr39tk supermutant was also confirmed by a drug sensitivity assay, in which the 50% inhibitory concentrations for thymine 1-beta-d-arabinofuranoside and bromovinyldeoxyuridine were found to be similar to those in nontransduced cells. In contrast, we found that HSV1-A167Ysr39tk could readily phosphorylate (3)H-GCV at levels similar to those of wild-type HSV1-tk and HSV1-A167Ytk but showed enhanced activity with (3)H-PCV in vitro and with (18)F-FHBG in vivo. CONCLUSION: We developed a new reporter gene, HSV1-A167Ysr39tk, which exhibits specificity and high phosphorylation activity for acycloguanosine derivatives. The resulting supermutant can be used for PET with (18)F-FHBG and suicidal gene therapy protocols with GCV in patients treated with pyrimidine-based cytotoxic drugs. PMID- 18413389 TI - 111In-labeled galectin-3-targeting peptide as a SPECT agent for imaging breast tumors. AB - Galectin-3 is a member of the galectin family of beta-galactoside-binding animal lectins. Galectin-3 is overexpressed in a wide range of neoplasms and is associated with tumor growth and metastases. Given this fact, radiolabeled galectin-3-targeting molecules may be useful for the noninvasive imaging of tumors expressing galectin-3, as well as for targeted radionuclide therapy. In this study, the tumor cell-targeting and SPECT properties of a galectin-3-avid peptide identified from bacteriophage display were evaluated in human breast carcinoma cells and in human breast tumor-bearing mice. METHODS: The galectin-3 avid peptide G3-C12 (ANTPCGPYTHDCPVKR) was synthesized with a Gly-Ser-Gly (GSG) linker at the amino terminus. After conjugation with 1,4,7,10-tetra azacyclododecane-N,N',N''N'''-tetraacetic acid (DOTA), the peptide was labeled with (111)In. The radiochemical purity and stability of the compound was assessed by high-performance liquid chromatography. MDA-MB-435 human breast carcinoma cells expressing galectin-3 were used to characterize the in vitro binding properties of the radiolabeled compound. SCID mice bearing MDA-MB-435 xenografts were used as an in vivo model for biodistribution and imaging studies with the (111)In-labeled peptide. RESULTS: (111)In-DOTA(GSG)-G3-C12 bound specifically to galectin-3-expressing MDA-MB-435 cells. The radiolabeled peptide was stable in serum and was found intact in excreted urine for at least 1 h. Competitive binding experiments indicated that the radiolabeled peptide exhibited an inhibitory concentration of 50% of 200.00+/-6.70 nM for cultured breast carcinoma cells. In vivo biodistribution studies revealed that tumor uptake was 1.2+/-0.24, 0.75+/-0.05, and 0.6+/-0.04 (mean +/- SD) percentage injected dose per gram at 30 min, 1.0 h, and 2.0 h after injection of the radiotracer, respectively. SPECT/CT studies with (111)In-DOTA(GSG)-G3-C12 showed excellent tumor uptake and contrast in the tumor-bearing mice. Specificity of peptide binding was demonstrated by successful blocking (52%) of in vivo tumor uptake of (111)In-DOTA(GSG)-G3-C12 in the presence of its nonradiolabeled counterpart at 2 h after injection. CONCLUSION: This study demonstrated the successful use of a new radiolabeled peptide for the noninvasive imaging of galectin-3-positive breast tumors. This peptide may be a promising candidate for future clinical applications. PMID- 18413390 TI - Risks associated with therapeutic 131I radiation exposure. PMID- 18413391 TI - Scintigraphy or multidetector CT angiography for suspected pulmonary embolism? PMID- 18413392 TI - Small-animal PET imaging of human epidermal growth factor receptor type 2 expression with site-specific 18F-labeled protein scaffold molecules. AB - Human epidermal growth factor receptor type 2 (HER2) is a well-established tumor biomarker that is overexpressed in a wide variety of cancers and that serves as a molecular target for therapeutic intervention. HER2 also serves as a prognostic indicator of patient survival and as a predictive marker of the response to antineoplastic therapy. The development of (18)F-labeled biomolecules for PET imaging of HER2 (HER2 PET) is very important because it may provide a powerful tool for the early detection of HER2-positive tumor recurrence and for the monitoring of HER2-based tumor treatment. METHODS: In this study, anti-HER2 monomeric and dimeric protein scaffold molecules [Z(HER2:477) and (Z(HER2:477))(2), respectively] were radiofluorinated at a reasonable radiochemical yield (13%-18%) by use of site-specific oxime chemistry. The resulting radiofluorinated protein scaffold molecules were then evaluated as potential molecular probes for small-animal HER2 PET by use of a SKOV3 tumor bearing mouse model. RESULTS: The 4-(18)F-fluorobenzaldehyde conjugated aminooxy protein scaffolds [(18)F-N-(4-fluorobenzylidene)oxime (FBO)-Z(HER2:477) and (18)F FBO-(Z(HER2:477))(2)] both displayed specific HER2-binding ability in vitro. Biodistribution and small-animal PET imaging studies further revealed that (18)F FBO-Z(HER2:477) showed rapid and high SKOV3 tumor accumulation and quick clearance from normal tissues, whereas (18)F-FBO-(Z(HER2:477))(2) showed poor in vivo performance (low tumor uptake and tumor-to-normal tissue ratios). The specificity of (18)F-FBO-Z(HER2:477) for SKOV3 tumors was confirmed by its lower uptake on pretreatment of tumor-bearing mice with the HER2-targeting agents Z(HER2) and trastuzumab. Moreover, small-animal PET imaging studies revealed that (18)F-FBO-Z(HER2:477) produced higher-quality tumor imaging than (18)F-FBO (Z(HER2:477))(2). (18)F-FBO-Z(HER2:477) could clearly identify HER2-positive tumors with good contrast. CONCLUSION: Overall, these data demonstrate that (18)F FBO-Z(HER2:477) is a promising PET probe for imaging HER2 expression in living mice. It has a high potential for translation to clinical applications. The radiofluorination method developed can also be used as a general strategy for the site-specific labeling of other proteins with (18)F. The protein scaffold molecules used here are attractive for the further development of PET probes for other molecular targets. PMID- 18413395 TI - DPA-714, a new translocator protein-specific ligand: synthesis, radiofluorination, and pharmacologic characterization. AB - The translocator protein (18 kDa) (TSPO), formerly known as the peripheral benzodiazepine receptor, is dramatically upregulated under pathologic conditions. Activated microglia are the main cell type expressing the TSPO at sites of central nervous system pathology. Radioligands for the TSPO can therefore measure active disease in the brain. This article details the synthesis, radiofluorination, and pharmacologic evaluation of a new TSPO-specific pyrazolopyrimidine, DPA-714. METHODS: The affinity of DPA-714 for the TSPO was measured in rat kidney membranes with (3)H-PK11195. The in vitro functional activity of DPA-714 was measured in a steroidogenic assay in which the ability of DPA-714 to increase pregnenolone synthesis was measured with rat C6 glioma cells. The radiofluorination of DPA-714 was achieved by nucleophilic (18)F-fluoride displacement of the tosylate precursor. (18)F-DPA-714 was assessed in rats harboring unilateral quinolinic acid (QA) lesions. In addition, pretreatment experiments were performed with PK11195 (5 mg/kg), DPA-714 (1 mg/kg), and DPA-713 (1 mg/kg). The in vivo binding and biodistribution of (18)F-DPA-714 were determined in a baboon with PET. Experiments involving presaturation with PK11195 (1.5 mg/kg) and displacement with DPA-714 (1 mg/kg) were conducted to evaluate the specificity of radioligand binding. RESULTS: In vitro binding studies revealed that DPA-714 displayed a high affinity for the TSPO (dissociation constant, 7.0 nM). DPA-714 stimulated pregnenolone synthesis at levels 80% above the baseline. (18)F-DPA-714 was prepared at a 16% radiochemical yield and a specific activity of 270 GBq/mumol. In rats harboring unilateral QA lesions, an 8 fold-higher level of uptake of (18)F-DPA-714 was observed in the ipsilateral striatum than in the contralateral striatum. Uptake in the ipsilateral striatum was shown to be selective because it was inhibited to the level in the contralateral striatum in the presence of PK11195, nonlabeled DPA-714, or DPA 713. PET studies demonstrated rapid penetration and good retention of (18)F-DPA 714 in the baboon brain. Pretreatment with PK11195 effectively inhibited the uptake of (18)F-DPA-714 in the whole brain, indicating its selective binding to the TSPO. The injection of nonlabeled DPA-714 20 min after the injection of (18)F DPA-714 resulted in radioligand washout, demonstrating the reversibility of (18)F DPA-714 binding. CONCLUSION: (18)F-DPA-714 is a specific radioligand for the TSPO, displaying promising in vivo properties and thus warranting further investigation. PMID- 18413396 TI - Prognostic value of 18F-fluoroethyl-L-tyrosine PET and MRI in small nonspecific incidental brain lesions. AB - Nonspecific incidental brain lesions (NILs) are being detected more frequently because of an increasing number of screening or research MRI scans of the brain, and their natural course is uncertain. METHODS: In a prospective cohort study starting in 1999, we determined the outcomes of patients with incidental, nonenhancing, supratentorial, lobar, and small-volume (<10 mL) lesions, depending on the findings of MRI and PET with the (18)F-labeled amino acid fluoroethyl-l tyrosine ((18)F-FET). Patients with seizures, focal neurologic deficits, signs of local or systemic infection or inflammation, known brain disease, or any kind of previous cerebral treatment were excluded. Finally, 21 patients were eligible. MRI was performed in 19 of these patients because of nonspecific symptoms (such as headaches, dizziness, or sudden deafness), whereas 2 patients were healthy volunteers in MRI studies. Clinical follow-up and MRI scans were obtained at 4- to 6-mo intervals, and follow-up ranged from 3 to 8.5 y. Mean lesion-to-brain (L/B) ratios of >or=1.6 on (18)F-FET PET were rated as positive. RESULTS: Four different outcome groups were identified. In group A, 5 NILs regressed or vanished completely. All of these lesions were circumscribed on MRI, and (18)F FET uptake was negative, with an L/B ratio of 1.2+/-0.2 (mean +/- SD). In group B, 10 NILs were stable, without growth. All of these lesions were circumscribed on MRI, and (18)F-FET uptake was negative (L/B ratio: 1.0+/-0.1). In group C, 2 NILs grew slowly over years, and an astrocytoma of World Health Organization (WHO) grade II was diagnosed after resection in each case. The lesions were circumscribed on MRI, and (18)F-FET uptake was negative (L/B ratios: 0.7 and 1.0). In group D, 4 NILs showed sudden and rapid growth, with clinical deterioration, and a high-grade glioma of WHO grade III or IV was diagnosed after resection in all cases. The lesions were diffuse on MRI, and (18)F-FET uptake was significantly increased (L/B ratio: 2.0+/-0.4) (P<0.01 for group D vs. group A or group B). CONCLUSION: For NILs, a circumscribed growth pattern on MRI and normal or low (18)F-FET uptake on PET are strong predictors for a benign course, with the eventual development of a low-grade glioma. In contrast, NILs with a diffuse growth pattern on MRI and increased (18)F-FET uptake indicate a high risk for the development of a high-grade glioma. PMID- 18413397 TI - Egg labeling for gastric-emptying studies. PMID- 18413398 TI - Uncertainties in internal dose calculations for radiopharmaceuticals. AB - This paper presents a systematic analysis of the inherent uncertainty in internal dose calculations for radiopharmaceuticals. A generic equation for internal dose is presented, and the uncertainty in each of the individual terms is analyzed, with the relative uncertainty of all terms compared. The combined uncertainties in most radiopharmaceutical dose estimates will be typically at least a factor of 2 and may be considerably greater. In therapy applications, if patient individualized absorbed doses are calculated, with attention being paid to accurate data gathering and analysis and measurement of individual organ volumes, many of the model-based uncertainties can be removed, and the total uncertainty in an individual dose estimate can be reduced to a value of perhaps +/-10%-20%. Radiation dose estimates for different diagnostic radiopharmaceuticals should be appreciated and considered, but small differences in dose estimates between radiopharmaceuticals should not be given too much importance when one is choosing radiopharmaceuticals for general clinical use. Diagnostic accuracy, ease of use, image quality, patient comfort, and other similar factors should predominate in the evaluation, with radiation dose being another issue considered while balancing risks and benefits appropriately. PMID- 18413399 TI - Therapeutic administration of 131I for differentiated thyroid cancer: radiation dose to ovaries and outcome of pregnancies. AB - Radiation is known to be mutagenic. The present study updates a 10-y-old study regarding pregnancy outcome and the health of offspring of women previously exposed to radioiodine ((131)I) during thyroid carcinoma treatment, by doubling the number of pregnancies that occurred after exposure. METHODS: Data on 2,673 pregnancies were obtained by interviewing female patients who were treated for thyroid carcinoma but had not received significant external radiation to the ovaries. RESULTS: The incidence of miscarriages was 10% before any treatment for thyroid cancer; this percentage increased after surgery for thyroid cancer, both before (20%) and after (19%) (131)I treatment, with no variation according to the cumulative dose. In contrast to previously reported data, miscarriages were not significantly more frequent in women treated with radioiodine during the year before conception, not even in women who had received more than 370 MBq during that year. The incidences of stillbirths, preterm births, low birth weight, congenital malformations, and death during the first year of life were not significantly different before and after (131)I therapy. The incidences of thyroid and nonthyroid cancers were similar in children born either before or after the mother's exposure to radioiodine. CONCLUSION: There is no evidence that exposure to radioiodine affects the outcomes of subsequent pregnancies and offspring. The question as to whether the incidences of malformations and thyroid and nonthyroid cancers are related to gonadal irradiation remains to be established. The doubling dose is still being heatedly debated, and the value of 1 Gy as the doubling dose in humans should be reevaluated. PMID- 18413400 TI - Comparison of 18F-FDG PET and MRI in assessment of uterine smooth muscle tumors. AB - The purpose of this study was to prospectively determine whether combined MRI and (18)F-FDG PET is more accurate than MRI in assessing nonbenign uterine smooth muscle tumors (USMTs). METHODS: Seventy patients (mean age, 49+/-10 y; range, 28 77 y) suspected of having nonbenign USMTs underwent both MRI and (18)F-FDG PET before surgery. Results were evaluated using receiver-operating-characteristic (ROC) analyses and the Cochran Q test. RESULTS: The area under the ROC curve for MRI with (18)F-FDG PET was significantly higher than that for MRI (0.97 vs. 0.89, P<0.05). Although multiple comparisons using the Cochran Q test were not significant, the sensitivity, specificity, and accuracy for MRI with (18)F-FDG PET with probable nonbenign USMT cases considered to be positive were higher than those for MRI (93.3% vs. 73.3%; 92.7% vs. 85.5%; and 92.9% vs. 82.9%, respectively). CONCLUSION: MRI with (18)F-FDG PET is useful in assessing nonbenign USMTs, as compared with MRI. PMID- 18413401 TI - Increased serotonin and dopamine transporter binding in psychotropic medication naive patients with generalized social anxiety disorder shown by 123I-beta-(4 iodophenyl)-tropane SPECT. AB - There is circumstantial evidence for the involvement of serotonergic and dopaminergic systems in the pathophysiology of social anxiety disorder. In the present study, using SPECT imaging we examined the (123)I-beta-(4-iodophenyl) tropane binding potential for the serotonin and dopamine transporters in patients with a generalized social anxiety disorder and in age- and sex-matched healthy controls. METHODS: Twelve psychotropic medication-naive patients with social anxiety disorder, generalized type (5 women and 7 men) and 12 sex- and age matched healthy controls were studied. Volumes of interest were constructed on MRI-coregistered SPECT scans. Binding ratios were compared using the Mann-Whitney U test. Possible correlations between binding patterns and symptomatology were assessed using the Spearman rank correlation coefficient. RESULTS: Significantly higher binding potentials were found for the serotonin in the left and right thalamus of patients. Patients had also a significantly higher binding potential for the dopamine transporter in the striatum. CONCLUSION: The present study provided direct evidence for abnormalities in both the dopaminergic and the serotonergic systems in patients with generalized social anxiety disorder. PMID- 18413402 TI - Metabolic imaging of cerebral gliomas: spatial correlation of changes in O-(2-18F fluoroethyl)-L-tyrosine PET and proton magnetic resonance spectroscopic imaging. AB - The aim of this study was to determine the spatial correlation of O-(2-(18)F fluoroethyl)-L-tyrosine ((18)F-FET) uptake and the concentrations of choline (Cho), creatine (Cr), and total N-acetylaspartate (tNAA) determined with proton magnetic resonance spectroscopic imaging ((1)H MRSI) in cerebral gliomas for the multimodal evaluation of metabolic changes. METHODS: (18)F-FET PET and 2 dimensional (1)H MRSI were performed in 15 patients with cerebral gliomas of World Health Organization (WHO) grades II-IV. PET and (1)H MRSI datasets were coregistered by use of mutual information. On the basis of their levels of (18)F FET uptake, 4 different areas in a tumor (maximum, strong, moderate, and low (18)F-FET uptake) were defined on PET slices as being congruent with the volume of interest in the (1)H MRSI experiment. (18)F-FET uptake in lesions was evaluated as tumor-to-brain ratios. Metabolite concentrations for Cho, Cr, and tNAA and Cho/tNAA ratios were computed for these 4 areas in the tumor and for the contralateral normal brain. RESULTS: In the area with maximum (18)F-FET uptake, the concentration of tNAA (R= -0.588) and the Cho/tNAA ratio (R=0.945) correlated significantly with (18)F-FET uptake. In the areas with strong and moderate (18)F FET uptake, only the Cho/tNAA ratios (R=0.811 and R=0.531, respectively) were significantly associated with amino acid transport. At low (18)F-FET uptake, analysis of the correlations of amino acid uptake and metabolite concentrations yielded a significant result only for the concentration of Cr (R=0.626). No correlation was found for metabolite concentrations determined with (1)H MRSI and (18)F-FET uptake in normal brain tissue. Maximum (18)F-FET uptake and the tNAA concentration were significantly different between gliomas of WHO grades II and IV, with P values of 0.032 and 0.016, respectively. CONCLUSION: High (18)F-FET uptake, which is indicative of tumor cell infiltration, associates with neuronal cell loss (tNAA) and changes in ratios between parameters representing membrane proliferation and those of neuronal loss (Cho/tNAA ratio), which can be measured by (1)H MRSI. The significant correlation coefficients detected for Cr in regions with low (18)F-FET uptake suggests an association between the mechanism governing amino acid transport and energy metabolism in areas that are infiltrated by tumor cells to a lesser extent. These findings motivate further research directed at investigating the potential of (1)H MRSI to define tumor boundaries in a manner analogous to that of amino acid PET. PMID- 18413403 TI - Comparison of the biodistribution and tumor targeting of two 99mTc-labeled anti EGFR nanobodies in mice, using pinhole SPECT/micro-CT. AB - Camelidae possess an unusual class of antibodies devoid of light chains. Nanobodies are intact antigen-binding fragments that are stable, easily generated against different targets, and fully functional. Their rapid clearance from the blood circulation favors their use as imaging agents. We compared the in vivo tumor uptake and biodistribution of 2 anti-epidermal growth factor receptor (anti EGFR) Nanobodies, (99m)Tc-7C12 and (99m)Tc-7D12. METHODS: Nanobodies were labeled via their hexahistidine tail with (99m)Tc-tricarbonyl ((99m)Tc(CO)(3)) generated from a kit. Mice bearing subcutaneous A431 (EGFR-positive) and R1M (EGFR negative) xenografts were intravenously injected with (99m)Tc-7C12 and (99m)Tc 7D12 on separate days. Pinhole SPECT/micro-CT images were acquired at 1 h after administration to assess noninvasively the biodistribution and tumor targeting of the labeled compounds. Pinhole SPECT and micro-CT images from the same mouse were automatically fused on the basis of a mathematic rigid-body-transformation algorithm using six (57)Co sources. Images were quantified, and tracer uptake was expressed as percentage injected activity per gram per cubic centimeter (%IA/cm(3)) of tissue. Ex vivo biodistribution of mice bearing A431 injected with either (99m)Tc-7C12 or (99m)Tc-7D12 was also assessed; activity in the tumor and organs was recorded and expressed as percentage injected activity per gram (%IA/g). RESULTS: Binding of both tracers was receptor-specific. Image analysis showed high and similar tumor uptake values for both (99m)Tc-7C12 and (99m)Tc 7D12 (4.55+/-0.24 %IA/cm(3) and 4.62+/-0.36 %IA/cm(3), respectively) in A431 xenografts, whereas the uptake in the negative tumor (R1M) was low (1.16+/-0.14 for (99m)Tc-7C12 and 1.49+/-0.60 for (99m)Tc-7D12). (99m)Tc-7C12 showed significantly higher kidney uptake (63.48+/-2.36 vs. 56.25+/-2.46 %IA/cm(3)) and lower liver uptake (2.55+/-0.26 vs. 4.88+/-0.86 %IA/cm(3)) than did (99m)Tc-7D12. The ex vivo analysis confirmed the image quantification with high tumor-to background ratio; however, (99m)Tc-7C12 showed higher tumor uptake (9.11+/-1.12 %IA/g) than did (99m)Tc-7D12 (6.09+/-0.77 %IA/g). (99m)Tc-7D12 demonstrated significantly higher blood activity than did (99m)Tc-7C12, but both showed short plasma half-lives (<10 min). CONCLUSION: The Nanobody fragments used here show high tumor uptake, low liver uptake, and rapid blood clearance. Nanobodies are promising probes for noninvasive radioimmunodetection of specific targets early after administration. On the basis of its favorable biodistribution, (99m)Tc-7C12 was selected for further studies. PMID- 18413404 TI - 203Pb-labeled alpha-melanocyte-stimulating hormone peptide as an imaging probe for melanoma detection. AB - Peptide-targeted alpha-therapy with 7.4 MBq of (212)Pb-[1,4,7,10 tetraazacyclododecane-1,4,7,10-tetraacetic acid]-ReO-[Cys(3,4,10),d Phe(7),Arg(11)]alpha-MSH(3-13) ((212)Pb-DOTA-Re(Arg(11))CCMSH) cured 45% of B16/F1 murine melanoma-bearing C57 mice in a 120-d study, highlighting its melanoma treatment potential. However, there is a need to develop an imaging surrogate for patient-specific dosimetry and to monitor the tumor response to (212)Pb-DOTA-Re(Arg(11))CCMSH therapy. The purpose of this study was to evaluate the potential of (203)Pb-DOTA-Re(Arg(11))CCMSH as a matched-pair SPECT agent for (212)Pb-DOTA-Re(Arg(11))CCMSH. METHODS: DOTA-Re(Arg(11))CCMSH was labeled with (203)Pb in 0.5 M NH(4)OAc buffer at pH 5.4. The internalization and efflux of (203)Pb-DOTA-Re(Arg(11))CCMSH were determined in B16/F1 melanoma cells. The pharmacokinetics of (203)Pb-DOTA-Re(Arg(11))CCMSH was examined in B16/F1 melanoma bearing C57 mice. A micro-SPECT/CT study was performed with (203)Pb-DOTA Re(Arg(11))CCMSH in a B16/F1 melanoma-bearing C57 mouse at 2 h after injection. RESULTS: (203)Pb-DOTA-Re(Arg(11))CCMSH was easily prepared in NH(4)OAc buffer and completely separated from the excess nonradiolabeled peptide by reversed-phase high-performance liquid chromatography (RP-HPLC). (203)Pb-DOTA-Re(Arg(11))CCMSH displayed fast internalization and extended retention in B16/F1 cells. Approximately 73% of (203)Pb-DOTA-Re(Arg(11))CCMSH activity internalized after a 20-min incubation at 25 degrees C. After incubation of the cells in culture medium for 20 min, 78% of internalized activity remained in the cells. (203)Pb DOTA-Re(Arg(11))CCMSH exhibited a biodistribution pattern similar to that of (212)Pb-DOTA-Re(Arg(11))CCMSH in B16/F1 melanoma-bearing mice. (203)Pb-DOTA Re(Arg(11))CCMSH exhibited a peak tumor uptake of 12.00+/-3.20 percentage injected dose per gram (%ID/g) at 1 h after injection. The tumor uptake gradually decreased to 3.43+/-1.12 %ID/g at 48 h after injection. (203)Pb-DOTA Re(Arg(11))CCMSH exhibited a peak tumor-to-kidney uptake ratio of 1.53 at 2 h after injection. The absorbed doses to the tumor and kidneys were 4.32 and 4.35 Gy, respectively, per 37 MBq. Whole-body clearance of (203)Pb-DOTA Re(Arg(11))CCMSH was fast, with approximately 89% of the injected activity cleared through the urinary system by 2 h after injection. (203)Pb showed 1.6-mm SPECT resolution, which was comparable to (99m)Tc. Melanoma lesions were visualized through SPECT/CT images of (203)Pb-DOTA-Re(Arg(11))CCMSH at 2 h after injection. CONCLUSION: (203)Pb-DOTA-Re(Arg(11))CCMSH exhibited favorable pharmacokinetic and tumor imaging properties, highlighting its potential as a matched-pair SPECT agent for (212)Pb-DOTA-Re(Arg(11))CCMSH melanoma treatment. PMID- 18413405 TI - Hydroxychloroquine: the cornerstone of lupus therapy. PMID- 18413406 TI - An update in pulmonary hypertension in systemic lupus erythematosus - do we need to know about it? AB - Pulmonary hypertension (PH) is a serious form of pulmonary complication that occurs less frequently in lupus than in other connective tissue diseases like scleroderma; however, it is likely that it is under-recognized in lupus. The symptoms of PH in lupus are non-specific (dyspnea, fatigue, impaired exercise tolerance) and can also be caused by other factors such as pleural or pericardial effusions, interstitial lung disease and many more, making it possible to miss the diagnosis. There are several potential causes of PH in lupus including thromboembolic disease, pulmonary vasculitis, and hypoxia and fibrosis from interstitial lung disease. Endothelin-1 is elevated in lupus and may be associated with PAH. In some studies, pulmonary arterial hypertension (PAH) has been found to be a major cause of mortality in lupus patients. Echocardiograms are a screening tool, but may yield false positives, and a right heart catheterization must be performed to confirm PAH. Early identification is important and can alter the natural history of this dangerous complication of lupus. Treatment of PAH associated with lupus includes standard PAH treatment as well as immunosuppression. PMID- 18413407 TI - Defining undifferentiated connective tissue diseases: a challenge for rheumatologists. PMID- 18413408 TI - Previous antimalarial therapy in patients diagnosed with lupus nephritis: influence on outcomes and survival. AB - The aim of this study was to analyze the effect of exposure to antimalarial drugs at diagnosis of lupus nephritis on the outcome of the disease, especially renal failure, comorbid processes, and survival. We analyzed a cohort of 206 consecutive patients with biopsy-proven lupus nephritis. Renal biopsies were categorized according to the classification proposed by the ISN/RPS in 2003. Exposure to antimalarial drugs (chloroquine and hydroxychloroquine) was defined as the use of these drugs before the diagnosis of lupus nephritis independent of dose and duration. Fifty-six (27%) patients had received antimalarials before the diagnosis of lupus nephritis. During the follow-up, these patients had a lower frequency of creatinine values >4 mg/dL (2% vs 11%, P = 0.029) and end-stage renal failure (2% vs 11%, P = 0.044) in comparison with those never treated with antimalarials. Patients exposed to antimalarials also had a lower frequency of hypertension (32% vs 50%, P = 0.027), infections (11% vs 29%, P = 0.006), and thrombotic events (5% vs 17%, P = 0.039). Twenty patients (10%) died during the study period. Patients exposed to antimalarials had a lower mortality rate at the end of the follow-up (2% vs 13% for those not exposed to antimalarials, P = 0.029). Multivariate analysis identified thrombosis and infections as statistically significant independent variables. Kaplan-Meier plots showed a lower rate of end-stage renal failure (log rank = 0.04) in patients exposed to antimalarials. In conclusion, exposure to antimalarials before the diagnosis of lupus nephritis was negatively associated with the development of renal failure, hypertension, thrombosis and infection, and with a better survival rate at the end of the follow-up. This, together with other published data, suggests that antimalarials should be considered a mandatory therapeutic option in all patients diagnosed with systemic lupus erythematosus. PMID- 18413409 TI - Quantitative and qualitative normal regulatory T cells are not capable of inducing suppression in SLE patients due to T-cell resistance. AB - Previous reports have suggested that regulatory T cells (Treg) are abnormal in patients with systemic lupus erythematosus (SLE). In the present work, we quantified CD4+FOXP3+ Treg cells in patients with SLE and found no quantitative alterations. However, we found a clear defect in suppression assays. Surprisingly, SLE-derived Treg cells exhibited a normal phenotype and functional capacity. Conversely, SLE-derived CD4+CD25(-) effector T cells resisted suppression by autologous and allogeneic regulatory cells. Our findings strongly suggest that the defect in T-cell suppression observed in SLE is because of effector cell resistance and not because of an abnormal regulatory function. PMID- 18413410 TI - High prevalence of infections in patients with systemic lupus erythematosus and pulmonary haemorrhage. AB - The main objective of this study is to describe the presence of infections in patients with pulmonary haemorrhage and systemic lupus erythematosus. Patients with systemic lupus erythematosus and pulmonary haemorrhage were thoroughly evaluated in the first 48 hours with imaging plus bronchoscopy and bronchoalveolar fluid analysis. If needed, videoassisted thoracoscopy and lung biopsy were performed too. In all, search for bacterial, mycobacterial and fungal infections proceeded. Appropriate blood, bronchoalveolar fluid and tissue cultures were taken. Patients were treated with antibiotics and corticosteroids in case of infection. Otherwise, they received initial intravenous methylprednsiolone pulses for 3 days as standard therapy for pulmonary haemorrhage in systemic lupus erythematosus. Additional treatment with immunosuppressives was further decided by the treating physicians. Fourteen events in 13 patients were evaluated. In eight events (57%), an infection was demonstrated. Aetiological agents included Pseudomonas sp. and Aspergillus fumigatus. Four patients died, three of them because of the pulmonary infection and one because of cerebral haemorrhage secondary to severe systemic hypertension, 48 hours after methylprednisolone treatment. Patients with systemic lupus erythematosus and pulmonary haemorrhage have a high prevalence of infections. The influence of pulmonary haemorrhage in the setting of systemic lupus erythematosus needs further study to establish adequate treatment and to reduce the high mortality of this complication. PMID- 18413411 TI - Anti-citrullinated peptide antibodies in lupus patients with or without deforming arthropathy. AB - The objective was to study the association of antibodies against cyclic citrullinated peptides (anti-CCP) in patients with lupus articular damage. We studied 34 systemic lupus erythematosus patients (30 women) with (n = 14) or without (n = 20) deforming arthropathy. Anti-DNA and arthritis were mandatory inclusion criteria for both groups. As controls, 34 patients with rheumatoid arthritis and nine patients with rheumatoid arthritis and systemic lupus erythematosus (rhupus) were included. Anti-CCP and rheumatoid factor were determined by ELISA and nephelometry respectively. All patients had recent x-ray films of the hands that were evaluated according to Sharp's method. Systemic lupus erythematosus patients had a mean 6.50 +/- 0.86 (SD, range 5-8) American College of Rheumatology (ACR) criteria, rheumatoid arthritis patients met 5.38 +/ 0.60 (range 4-6) ACR criteria for rheumatoid arthritis and rhupus patients had 5.78 +/- 0.44 (range 5-6) criteria for rheumatoid arthritis and 5.11 +/- 0.78 (range 4-6) for systemic lupus erythematosus. Systemic lupus erythematosus patients, with or without deforming arthropathy, had normal serum anti-CCP concentrations. In contrast, rheumatoid arthritis and rhupus patients had 30- and 23-fold higher than normal amounts of anti-CCP (p < 0.001, both comparisons). Rheumatoid arthritis (97%) and rhupus (100%) patients were more frequently positive for anti-CCP than SLE patients with (7%) or without (5%) deforming arthropathy (p < 0.001, both comparisons). Patients with lupus deforming arthropathy were more frequently positive for rheumatoid factor (65%) than patients with non-deforming arthritis (15%) (p = 0.005). Patients with lupus deforming arthropathy had similar frequency of erosions and mean Sharp's score than rhupus patients. Anti-CCP antibodies do not associate with lupus arthropathy, whether deforming, non-deforming or erosive. PMID- 18413412 TI - Rapamycin prevents the development of nephritis in lupus-prone NZB/W F1 mice. AB - Rapamycin is a potent immunosuppressive drug currently used mainly for rejection prophylaxis in renal transplantation. The aim of this study was to determine the effect of rapamycin treatment on the development of nephritis in lupus-prone New Zealand Black/White F1 (NZB/W F1) mice. Twelve-week-old female NZB/W F1 mice were treated with rapamycin (3 mg/kg body weight) or saline once daily by oral gavage for 20 weeks. The severity of nephritis was assessed by clinical and biochemical parameters, renal histology, immunohistochemistry and gene expression studies. Rapamycin treatment markedly reduced proteinuria, improved renal function, decreased serum anti-double stranded DNA antibody levels and diminished splenomegaly. Kidney sections from saline-treated mice showed marked mesangial proliferation, tubular dilation with protein cast deposition and interstitial inflammatory cell infiltration. Rapamycin-treated mice had near normal renal histology, with marked reduction in glomerular immune deposition and the infiltration by T cells, B cells and macrophages. Rapamycin treatment was associated with down-regulation of intra-renal expression of monocyte chemoattractant protein-1 (MCP-1) mRNA and protein. We conclude that rapamycin is highly effective in preventing the development of nephritis in NZB/W F1 mice. The beneficial effects of rapamycin are mediated through inhibition of lymphoproliferation and reduced MCP-1 expression. PMID- 18413414 TI - Prognostic factors of lupus myelopathy. AB - Myelopathy is a rare but severe neurological manifestation of systemic lupus erythematosus (SLE) with a high morbidity. The factors that contribute to prognosis are unknown. In this study, 14 patients with lupus myelopathy (LM) from our centre were retrospectively studied. Another 23 patients with other neuropsychiatric SLE (NPSLE) features were enrolled as a comparison group. The morbidity of LM was evaluated by the ASIA Impairment Scale. The clinical and serological characteristics and prognostic factors for LM were investigated. The age, gender, duration of SLE, non-CNS disease activity and autoantibody profile in patients with LM was not different in the NPSLE cohort. A relatively low prevalence of anti-phospholipid antibodies (aPL) in LM sera compared to NPSLE (28.6% vs 52.2%, P = 0.19) was observed. Longitudinal lesion detected by magnetic resonance imaging (MRI) was identified in 33.3% of patients with LM, whereas 50% showed focal speckle-like lesions. The morbidity of LM is 50%. Muscle strength of grade 3 or higher on admission was a strong indicator for a better prognosis (P = 0.006), whereas other parameters including longitudinal lesion, sensory deficit level, disease activity and aPL did not discriminate good from poor outcome in LM. Early aggressive immunosuppressive therapy (within 2 weeks of onset of myelopathy) tend to associate with a favourable outcome (P = 0.07). PMID- 18413413 TI - Adolescent onset of lupus results in more aggressive disease and worse outcomes: results of a nested matched case-control study within LUMINA, a multiethnic US cohort (LUMINA LVII). AB - The objective of this study is to examine the clinical features and outcomes of patients with systemic lupus erythematosus (SLE) whose disease began in adolescence [juvenile-onset SLE (jSLE)] compared with adult-onset patients [adult onset SLE (aSLE)] from a large multiethnic cohort. Systemic lupus erythematosus patients of African-American, Caucasian, or Hispanic ethnicity and >or=1 year follow-up were studied in two groups: jSLE (diagnosed at 10 mg/day (OR = 3.69; 95% CI 1.22-11.11) were associated with metabolic syndrome. In conclusion, older age, low socioeconomic status, lack of exercise, thrombocytopenia, increased erythrocyte sedimentation rate , higher disease activity, and prednisone >10 mg/day were independently associated with metabolic syndrome in patients with systemic lupus erythematosus from Puerto Rico. PMID- 18413420 TI - The effect of sports injury on insulin-like growth factor-I and type 3 procollagen: implications for detection of growth hormone abuse in athletes. AB - CONTEXT: A method to detect exogenously administered growth hormone (GH) based on the measurement of two GH-dependent markers, IGF-I and type 3 procollagen (P-III P) has been proposed. Skeletal or soft tissue injury may alter these markers. Elevations in either of these proteins after injury might lead to a false accusation of doping with GH. OBJECTIVE: The objective of the study was to assess the effect of musculoskeletal or soft tissue injury on IGF-I and P-III-P concentrations in amateur and elite athletes and assess the effect of injury on the proposed GH detection method. DESIGN: This was a longitudinal observational study after sporting injury. SETTING: The study was conducted at Southampton General Hospital and British Olympic Medical Centre. SUBJECTS: Subjects included elite and amateur athletes after an injury. INTERVENTION: Interventions included measurement of IGF-I and P-III-P and application of the GH-2000 discriminant function score up to 84 d after an injury as well as classification of injury by type and severity. OUTCOME MEASURES: IGF-I and P-III-P concentration and ability to detect GH abuse in athletes without the risk of false accusation because of an injury were measured. RESULTS: There was no change in IGF-I concentration after an injury. By contrast, P-III-P concentrations rose by 41.1 +/- 16.6%, reaching a peak around 14 d after an injury. The rise in P-III-P varied according to injury type and severity. This rise had a trivial effect on the GH-2000 discriminant function score, and no subject reached the threshold needed for a doping offense. CONCLUSIONS: Although there was a rise in P-III-P after injury, this was insufficient to invalidate the GH-2000 detection method based on IGF-I and P-III P concentrations. PMID- 18413421 TI - Aerobic capacity and growth hormone deficiency after traumatic brain injury. AB - CONTEXT: GH deficiency occurs in approximately 20% of all individuals who suffer from a moderate to severe traumatic brain injury. OBJECTIVE: This study determined whether GH deficiency secondary to traumatic brain injury had an effect on aerobic capacity. DESIGN: Subjects were screened for GH deficiency by the glucagon stimulation test and performed a maximal treadmill exercise test. SETTING: Patients were studied in the postacute recovery phase after traumatic brain injury. PARTICIPANTS: Thirty-five individuals were studied. Groups were formed as follows: normal GH axis, greater than 8 ng/ml response (n = 12); insufficient, GH 3-8 ng/ml response (n = 11); and deficient, less than 3 ng/ml response (n = 12). INTERVENTION: There was no intervention. MAIN OUTCOME MEASURE: Aerobic capacity was assessed by measuring expired gases during a graded treadmill exercise test. One-way and two-way ANOVAs were carried out on all peak and submaximal cardiorespiratory variables, respectively. Appropriate post hoc comparisons followed as necessary. RESULTS: Significantly higher peak oxygen consumption was found in traumatic brain injury subjects with GH normal vs. GH insufficient and deficient [26.4 +/- 6.9, 20.8 +/- 4.6, and 19.7 +/- 5.0, respectively (P < 0.05)]. Submaximal oxygen consumption was significantly higher in the GH normal group. All other variables were statistically similar. CONCLUSIONS: This study shows that individuals with traumatic brain injury with normal GH secretion have below normal aerobic capacity and those patients who have GH insufficiency/deficiency are further deconditioned. Studies of GH replacement in these subjects should be conducted to assess whether GH therapy can improve cardiorespiratory fitness and prevent secondary disability. PMID- 18413422 TI - Iodine prophylaxis using iodized salt and risk of maternal thyroid failure in conditions of mild iodine deficiency. AB - CONTEXT: Mild to moderate iodine deficiency during pregnancy can cause transient maternal hypothyroidism and impaired mental development of the progeny. These unfavorable effects are preventable by iodine supplementation. In Europe, however, less than 50% pregnant women receive iodine-containing supplements, thus representing dietary iodized salt the only carrier of iodine for most women in this life stage. OBJECTIVE/DESIGN: This longitudinal study is aimed to investigate the effects of long-term iodized salt consumption on maternal thyroid function during gestation. PARTICIPANTS/OUTCOME MEASURES: We prospectively evaluated thyroid function in 100 consecutive thyroperoxidase antibody-negative pregnant women from a mildly iodine-deficient area. Sixty-two women who had regularly used iodized salt for at least 2 yr prior to becoming pregnant and 38 who commenced iodized salt consumption upon becoming pregnant were classified as long-term (LT) and short-term (ST) iodine supplemented, respectively. RESULTS: Long-term iodized salt consumption resulted in a very low prevalence of maternal thyroid failure (MTF) in LT women. Conversely, short-term iodine prophylaxis does not seem to protect against the risk of MTF, the prevalence of which was almost 6 fold higher in ST than LT women (36.8% vs. 6.4%; chi(2) 14.7, P < 0.0005; relative risk 5.7, 95% confidence interval 2.03-16.08, P < 0.001). The relative risk reduction amounted to 82.5%, this measure indicating the extent to which long-term iodine prophylaxis using iodized salt would reduce the risk of MTF in ST women. CONCLUSIONS: Prolonged iodized salt significantly improves maternal thyroid economy and reduces the risk of maternal thyroid insufficiency during gestation, probably because of a nearly restoring intrathyroidal iodine stores. PMID- 18413423 TI - Male hormonal contraception: a double-blind, placebo-controlled study. AB - BACKGROUND: This study was performed to assess spermatogenesis suppression and safety of a new combination of an etonogestrel (ENG) implant combined with testosterone undecanoate (TU) injections for male contraception. This is the first large placebo-controlled study for male hormonal contraception. DESIGN AND STUDY SUBJECTS: In this double-blind, multicenter study, we randomly assigned 354 healthy men to receive either a low- or high-release ENG implant sc combined with im TU injections (750 mg every 10 or 12 wk or 1000 mg every 12 wk) or placebo implant and injections. Treatment duration was 42 or 44 wk and posttreatment follow-up at least 24 wk. RESULTS: Overall, spermatogenesis was suppressed to 1 million/ml or less at wk 16 in 89% of men, with approximately 94% in two high release ENG groups. Suppression was maintained up to the end of the treatment period in 91% of men. For all men who completed the treatment period, 3% never achieved 1 million/ml or less. Median recovery time to a sperm concentration above 20 million/ml was 15 wk (mean 17 wk, 95% confidence interval 16-18 wk). Treatment was well tolerated. As compared with the placebo group, more men in the active treatment groups reported adverse events such as weight gain, mood changes, acne, sweating, or libido change. For both spermatogenesis suppression and safety, differences were small between the active treatment groups. CONCLUSIONS: The combination of an ENG implant with TU injections is a well tolerated male hormonal method, providing effective and reversible suppression of spermatogenesis. Although the results are good, there is still room for improvement, possibly by adjusting the dose regimen or changing the mode of application. PMID- 18413424 TI - The role of stereotactic radiotherapy in patients with growth hormone-secreting pituitary adenoma. AB - CONTEXT: Single-session stereotactic radiotherapy (SR) may be a potential adjuvant treatment in acromegaly. OBJECTIVE: We analyzed the safety and efficacy of SR in patients who had previously received maximal surgical debulking at our center. DESIGN: The study was a retrospective analysis of hormonal, radiological, and ophthalmologic data collected in a predefined protocol from 1994 through 2006. SETTING: The study was performed at a university hospital. PATIENTS: Eighty three acromegalic patients, 52 women and 31 men, with a mean age of 42.6 +/- 1.2 yr, participated in the study. The median follow-up was 69 months (interquartile range 44-107 months). INTERVENTION: The patients were treated with SR for residual or recurrent GH-secreting adenoma. MAIN OUTCOME MEASURE: Normalization of age- and sex-adjusted IGF-I levels together with a basal GH level below 2.5 microg/liter without concomitant GH-suppressive drugs was the goal of therapy. RESULTS: Fifty patients (60.2%) reached the main outcome of the study. The rate of remission was 52.6% at 5 yr [95% confidence interval (CI) 40.6-64.6%]. Another 13 patients (15.7%), who were resistant to somatostatin analogs, were in remission after SR. Multivariate analysis showed that low basal GH and IGF-I levels were associated with a favorable outcome. No serious side effects occurred after SR. The 5-yr cumulative risk of new onset hypogonadism, hypothyroidism, or hypoadrenalism was 3.6% (95% CI 0-8.6%), 3.3% (95% CI 0-7.7%), and 4.9% (95% CI 0 10.4%), respectively. CONCLUSION: In a highly selected group of acromegalic patients, SR treatment had good efficacy and safety. This may lead to reconsider the role of SR in the therapeutic algorithm of acromegaly. PMID- 18413425 TI - Role of growth hormone in regulating lipolysis, proteolysis, and hepatic glucose production during fasting. AB - CONTEXT: Fasting is associated with suppressed insulin and augmented GH secretion. The involvement of each mechanism in the regulation of fuel mobilization during fasting is unknown. OBJECTIVE: To ascertain the role of GH in the regulation of the rates of lipolysis, proteolysis, and hepatic glucose production (HGP) during the physiological daily feed/fast cycle and after 2 d of complete fasting, we used a model of selective GH suppression by the administration of GHRH receptor antagonist (GHRH-A). DESIGN AND SETTING: We conducted an open label in-patient study in the General Clinical Research Center at the University of Michigan. PARTICIPANTS: Six healthy, nonobese volunteers participated. MAIN OUTCOME MEASURES: We assessed 24-h plasma GH concentration and rates of lipolysis, proteolysis, and HGP using stable isotope techniques after an overnight fast and after 2 d of fasting. RESULTS: GHRH-A suppressed plasma GH by about 65% during the fed state (P = 0.015) but did not alter the rates of lipolysis, proteolysis, or HGP. Fasting for 2 d suppressed plasma insulin concentration by about 80% and elevated plasma GH about 4-fold (both P < 0.01). This was accompanied by a doubling in the rate of lipolysis, an approximately 40% increase in proteolysis, and an approximately 30% decline in HGP (all P < 0.05). Preventing the fasting-induced increase in GH with GHRH-A largely abolished the increase in the rate of lipolysis. GHRH-A also augmented the fasting-induced reduction in HGP but did not alter proteolysis. CONCLUSIONS: Endogenous GH plays a very limited metabolic role during the daily feed/fast cycle but is essential for the increased lipolytic rate found with more prolonged fasting. PMID- 18413426 TI - Treatment of hypovitaminosis D in infants and toddlers. AB - CONTEXT: Hypovitaminosis D appears to be on the rise in young children, with implications for skeletal and overall health. OBJECTIVE: The objective of the study was to compare the safety and efficacy of vitamin D2 daily, vitamin D2 weekly, and vitamin D3 daily, combined with supplemental calcium, in raising serum 25-hydroxyvitamin D [25(OH)D] and lowering PTH concentrations. DESIGN: This was a 6-wk randomized controlled trial. SETTING: The study was conducted at an urban pediatric clinic in Boston. SUBJECTS: Forty otherwise healthy infants and toddlers with hypovitaminosis D [25(OH)D < 20 ng/ml] participated in the study. INTERVENTIONS: Participants were assigned to one of three regimens: 2,000 IU oral vitamin D2 daily, 50,000 IU vitamin D2 weekly, or 2,000 IU vitamin D3 daily. Each was also prescribed elemental calcium (50 mg/kg.d). Infants received treatment for 6 wk. MAIN OUTCOME MEASURES: Before and after treatment, serum measurements of 25(OH)D, PTH, calcium, and alkaline phosphatase were taken. RESULTS: All treatments approximately tripled the 25(OH)D concentration. Preplanned comparisons were nonsignificant: daily vitamin D2 vs. weekly vitamin D2 (12% difference in effect, P = 0.66) and daily D2 vs. daily D3 (7%, P = 0.82). The mean serum calcium change was small and similar in the three groups. There was no significant difference in PTH suppression. CONCLUSIONS: Short-term vitamin D2 2,000 IU daily, vitamin D2 50,000 IU weekly, or vitamin D3 2,000 IU daily yield equivalent outcomes in the treatment of hypovitaminosis D among young children. Therefore, pediatric providers can individualize the treatment regimen for a given patient to ensure compliance, given that no difference in efficacy or safety was noted among these three common treatment regimens. PMID- 18413428 TI - Reductions in basal metabolic rate and physical activity contribute to hypothalamic obesity. AB - CONTEXT: Obesity after hypothalamic damage is often severe and resistant to lifestyle changes. It is postulated that differences in basal metabolic rate (BMR) and physical activity may contribute to hypothalamic obesity (HO). OBJECTIVE: Our objective was to investigate the role of energy expenditure, BMR, and physical activity in the etiology of hypothalamic obesity. DESIGN: This was a cross-sectional study of three groups of children: those with HO, congenital hypopituitarism (CH), and simple obesity (SO). RESULTS: A total of 47 children (HO = 18, CH = 13, and SO = 16) had BMR measured, using indirect calorimetry (Deltatrac II). A lower BMR was seen in the HO group, which remained even after adjusting for lean mass. Physical activity, assessed using triaxial accelerometry, demonstrated longer activity periods in the HO group, although the degree of activity was reduced. No significant differences were seen in calorie intake. CONCLUSION: Energy expenditure, rather than energy intake, has a greater role in the development of obesity after cranial tumor therapy. Reductions in BMR and physical activity, leading to a positive energy balance and weight gain despite an age-appropriate calorie intake, may contribute to hypothalamic obesity. PMID- 18413427 TI - Treatment of adrenocorticotropin-dependent Cushing's syndrome: a consensus statement. AB - OBJECTIVE: Our objective was to evaluate the published literature and reach a consensus on the treatment of patients with ACTH-dependent Cushing's syndrome, because there is no recent consensus on the management of this rare disorder. PARTICIPANTS: Thirty-two leading endocrinologists, clinicians, and neurosurgeons with specific expertise in the management of ACTH-dependent Cushing's syndrome representing nine countries were chosen to address 1) criteria for cure and remission of this disorder, 2) surgical treatment of Cushing's disease, 3) therapeutic options in the event of persistent disease after transsphenoidal surgery, 4) medical therapy of Cushing's disease, and 5) management of ectopic ACTH syndrome, Nelson's syndrome, and special patient populations. EVIDENCE: Participants presented published scientific data, which formed the basis of the recommendations. Opinion shared by a majority of experts was used where strong evidence was lacking. CONSENSUS PROCESS: Participants met for 2 d, during which there were four chaired sessions of presentations, followed by general discussion where a consensus was reached. The consensus statement was prepared by a steering committee and was then reviewed by all authors, with suggestions incorporated if agreed upon by the majority. CONCLUSIONS: ACTH-dependent Cushing's syndrome is a heterogeneous disorder requiring a multidisciplinary and individualized approach to patient management. Generally, the treatment of choice for ACTH-dependent Cushing's syndrome is curative surgery with selective pituitary or ectopic corticotroph tumor resection. Second-line treatments include more radical surgery, radiation therapy (for Cushing's disease), medical therapy, and bilateral adrenalectomy. Because of the significant morbidity of Cushing's syndrome, early diagnosis and prompt therapy are warranted. PMID- 18413430 TI - Low circulating insulin-like growth factor I bioactivity in elderly men is associated with increased mortality. AB - CONTEXT: Low IGF-I signaling activity prolongs lifespan in certain animal models, but the precise role of IGF-I in human survival remains controversial. The IGF-I kinase receptor activation assay is a novel method for measuring IGF-I bioactivity in human serum. We speculated that determination of circulating IGF-I bioactivity is more informative than levels of immunoreactive IGF-I. OBJECTIVE: Our objective was to study IGF-I bioactivity in relation to human survival. DESIGN, SETTING, AND STUDY PARTICIPANTS: We conducted a prospective observational study at a clinical research center at a university hospital of 376 healthy elderly men (aged 73-94 yr). MAIN OUTCOME MEASURES: IGF-I bioactivity was determined by the IGF-I kinase receptor activation assay. Total and free IGF-I were determined by IGF-I immunoassays. Mortality was registered during follow-up (mean 82 months). RESULTS: During the follow-up period of 8.6 yr, 170 men (45%) died. Survival of subjects in the highest quartile of IGF-I bioactivity was significantly better than in the lowest quartile, both in the total study group [hazard ratio (HR) = 1.8; 95% confidence interval (95% CI) = 1.2-2.8; P = 0.01] as well as in subgroups having a medical history of cardiovascular disease (HR = 2.4; 95% CI = 1.3-4.3; P = 0.003) or a high inflammatory risk profile (HR = 2.3; 95% CI = 1.2-4.5; P = 0.01). Significant relationships were not observed for total or free IGF-I. CONCLUSION: Our study suggests that a relatively high circulating IGF-I bioactivity in elderly men is associated with extended survival and with reduced cardiovascular risk. PMID- 18413429 TI - The association of testosterone levels with overall sleep quality, sleep architecture, and sleep-disordered breathing. AB - CONTEXT: Little is known about the association of low endogenous testosterone levels and abnormal sleep patterns in older men, although pharmacological doses of testosterone are associated with increased severity of sleep apnea and other sleep disturbances. OBJECTIVE: The objective of the study was to examine the association between serum testosterone levels with objectively measured sleep characteristics. DESIGN: This was a cohort study. SETTING: Community-dwelling men aged 65 yr or older from six clinical centers in the United States participated in the study. PARTICIPANTS AND MAIN OUTCOME MEASURES: A total of 1312 men had baseline total testosterone levels measured in 2000-2002, followed 3.4 yr later by 72-h (minimum) actigraphy and one-night in-home polysomnography to assess sleep duration, sleep fragmentation, and sleep apnea. Analyses were performed by quartile of total testosterone and categorically defined low vs. higher total testosterone (<250 ng/dl vs. > or =250 ng/dl). Lifestyle and body size were covariates. RESULTS: Total testosterone levels were unrelated to age or duration of sleep. Men with lower testosterone levels had lower sleep efficiency, with increased nocturnal awakenings and less time in slow-wave sleep as well as a higher apnea-hypopnea index and more sleep time with O(2) saturation levels below 90%. Low testosterone levels were associated with overweight, and all significant associations were attenuated or absent after adjusting for body mass index or waist circumference. In a post hoc analysis in men with higher body mass index (>27 kg/m2), testosterone was significantly associated with more periods awake after sleep onset and lower sleep efficiency. CONCLUSION: Low total testosterone levels are associated with less healthy sleep in older men. This association is largely explained by adiposity. Clinical trials are necessary to determine whether body weight acts directly or indirectly (via low testosterone) in the causal pathway for sleep-disordered breathing in older men. PMID- 18413431 TI - The HLA-DRB1*1501 allele is prevalent among Japanese patients with anti glomerular basement membrane antibody-mediated disease. AB - BACKGROUND: We aimed to clarify the relationship between HLA-DRB1(*)1501 and anti glomerular basement membrane (GBM) antibody-mediated disease in Japanese patients. MATERIALS: Samples were collected from 16 anti-GBM antibody-positive patients who were admitted to our department or related hospitals from December 1990 to October 2005. We analysed clinical and laboratory data, kidney biopsy findings, and the HLA-DR phenotypes and HLA-DRB1 alleles of the patients. RESULTS: Among the 16 patients, 15 had HLA-DR15 [the phenotype frequency (PF) was 93.8%], 7 were positive for DR4 (the PF was 43.8%) and 5 were positive for DR9 (the PF was 31.3%). The allele frequency of HLA-DRB1(*)1501 was 46.4% (13/28), which was significantly different from Japanese controls (11.6%) (P < 0.001). In contrast, the frequency of HLA-DRB1(*)1502 was not different from controls (0/28). The odds ratio of HLA-DRB1(*)1501 in these patients was 6.4 (95% CI: 2.4 16.5). CONCLUSION: The present study demonstrated that Japanese patients with anti-GBM antibody-mediated disease are very likely to carry the HLA-DRB1(*)1501 but not the HLA-DRB1(*)1502 allele. PMID- 18413432 TI - Class-related health inequalities are not larger in the East: a comparison of four European regions using the new European socioeconomic classification. AB - BACKGROUND: The article investigates whether people in Eastern Europe have larger health inequalities than their counterparts in three West European regions (North, Central and the South). METHODS: Data were obtained for 63,754 individuals in 23 countries from the first (2002) and second (2004) waves of the European Social Survey. The health outcomes were self-reported limiting longstanding illness and fair/poor general health. Occupational class was defined according to the European Socioeconomic Classification (ESeC). The magnitude of absolute and relative inequalities according to nine occupational classes for men and women separately were identified, analysed and compared in all four regions of Europe. RESULTS: For both sexes and within all European regions, the higher and lower professionals, self-employed and higher service workers reported fewer cases of ill health than other occupational classes. In contrast, lower technical and routine workers reported the poorest health, excluding the relatively small number of farmers. Income and education did not explain more, or less, of the class-related health inequalities in the East compared with the other regions. CONCLUSIONS: Little evidence was found for the hypothesis that East European countries have larger class-related health inequalities than other European regions. People's income and educational attainment both contribute to occupational health inequalities in the East as well as in the West. PMID- 18413434 TI - The effect of eradicating poverty on childhood unintentional injury mortality in New Zealand: a cohort study with counterfactual modelling. AB - BACKGROUND: The aim of this study was to examine the effect of household income on unintentional injury mortality in children and to model the potential impact of eradicating income poverty as an injury prevention strategy. METHODS: A national retrospective cohort study linking census to mortality records carried out in New Zealand during a 3-year period following the 1991 census and including children aged 0-14 years on census night. The main outcome measures are odds ratios (ORs) for unintentional injury death by equivalised household income category and proportional reductions (population-attributable risk) in unintentional injury mortality from modelled scenarios of nil poverty. RESULTS: One-third of children lived in households earning less than 60% of the national median household income. Age-adjusted odds of death from unintentional injury were higher for children from any income category compared with the highest, and were most elevated for children from households earning less than 40% of the national median income (OR 2.81, 95% CI 1.73 to 4.55). Adjusting for ethnicity, household education, family status and labour force status halved the effect size (OR 1.83, 1.02 to 3.28). Thirty per cent of injury mortality was attributable to low or middle household income using the highest income category as reference. Altering the income distribution to eradicate poverty, defined by a threshold of 50% or 60% of the national median income, reduced injury mortality in this model by a magnitude of 3.3% to 6.6%. CONCLUSIONS: Household income is related to a child's risk of death from unintentional injury independent of measured confounders. Most deaths attributable to low income occur among households that are not defined as "in poverty". The elimination of poverty may reduce childhood unintentional injury mortality by 3.3% to 6.6%. PMID- 18413433 TI - Socioeconomic position and hysterectomy: a cross-cohort comparison of women in Australia and Great Britain. AB - OBJECTIVES: To examine the associations between indicators of socioeconomic position (SEP) and hysterectomy in two Australian and two British cohorts. STUDY POPULATION: Women participating in the Australian Longitudinal Study on Women's Health (ALSWH), born 1921-1926 and 1946-1951, and two cohorts of British women, the British Women's Heart and Health Study and the MRC National Survey of Health and Development, born at similar times (1920 to 1939 and 1946, respectively) and surveyed at similar ages to the ALSWH cohorts. METHODS: Relative indices of inequality were derived for own and head of household occupational class, educational level attained and age at leaving school. Logistic regression was used to test the associations between these indicators of SEP and self-reported hysterectomy and/or oophorectomy. RESULTS: Inverse associations between indicators of SEP and hysterectomy were found in both the Australian and British cohorts of women born in 1946 or later. There was also evidence of an inverse association between education and hysterectomy in the older Australian cohort. However, the associations in this older cohort were weaker than those found in the mid-aged Australian cohort. In the older British cohort, born in the 1920s and 1930s, little evidence of association between SEP in adulthood and hysterectomy was found. CONCLUSIONS: These results suggest that inverse associations between indicators of SEP and hysterectomy are stronger in younger than in older cohorts in both Australia and Great Britain. They provide further evidence of the dynamic nature of the association between indicators of SEP and hysterectomy. PMID- 18413435 TI - Dentine lead levels in childhood and criminal behaviour in late adolescence and early adulthood. AB - INTRODUCTION: There have been claims in the literature that lead exposure makes a strong contribution to criminal behaviour. OBJECTIVES: To examine linkages between lead exposure in childhood and crime in late adolescence/early adulthood to (a) determine whether higher levels of lead exposure were associated with increased levels of criminal behaviour and (b) estimate the extent to which lead exposure was responsible for increases in criminal behaviour. METHODS: Negative binomial regression models were fitted using data from a longitudinal birth cohort of New Zealand-born children studied from birth to age 21. RESULTS: There were statistically significant (p<0.05) bivariate associations between dentine lead levels at ages 6-9 and (a) officially recorded violence/property convictions (ages 14-21) and (b) self-reported violent/property offending (ages 14-21). The mean rate of convictions was 1.89 (SD 6.86) and the mean rate of offences was 15.24 (SD 49.24) for those with the highest level of exposure. Those with the lowest level of exposure had a mean rate of convictions of 0.0, and a mean rate of offending of 1.97 (SD 6.34). Adjustment for confounding factors reduced the magnitude of these associations, but the associations remained statistically significant. Further analyses suggested that the associations were largely explained by the linkages between lead exposure and educational underachievement. Lead exposure accounted for less than 1% of the variance in crime. CONCLUSIONS: These results suggest that, although lead exposure was associated with criminal behaviour, the associations were somewhat weak, and were largely explained by linkages between lead exposure and educational underachievement. PMID- 18413436 TI - High agreement on family affluence between children's and parents' reports: international study of 11-year-old children. AB - OBJECTIVE: To examine the agreement between parents' and children's reports on four items of family affluence: number of cars, own bedroom, number of family holidays and number of computers, and to analyse predictors of disagreement. DESIGN: Cross-sectional child-parent validation study of selected items from an internationally standardised questionnaire. SETTING: Survey conducted in schools in Denmark, Germany, Hungary, Italy, Poland and Scotland. PARTICIPANTS: 972 11 year-old children and their parents responded to the questionnaires. RESULTS: The child item response rates were high (above 93%). The per cent agreement was low for holidays spent with family (52.5%), but high for the other three items of family affluence (76.2-88.1%). The kappa coefficients were good or excellent for all items (between 0.41 and 0.74) and the gamma coefficients were strong for all items (between 0.56 and 0.96). Children from single-parent families were more likely to over-report family affluence (OR 2.67; CI 1.83 to 3.89). CONCLUSIONS: Young adolescents' self-reports of family affluence are fairly valid across the six countries. This finding suggests that the variables measured can be used in epidemiological studies that aim at ranking children according to socioeconomic position. PMID- 18413437 TI - The effects of workplace downsizing on cause-specific mortality: a register-based follow-up study of Finnish men and women remaining in employment. AB - BACKGROUND: Experience of workplace downsizing (ie reduction in personnel) is common and may constitute a threat to public health in working populations. This study aimed to determine whether downsizing was associated with increased mortality among those remaining in the downsized workplaces. METHODS: Prospective population registration data containing detailed socioeconomic and demographic information on 85 833 Finnish employees aged 35-64 years at the beginning of 1994 or 1993 followed up for cause-specific mortality for 8 years. One-year changes in workplace staffing levels were obtained from Statistics Finland records on workplaces. RESULTS: There was no association between downsizing on any level (a 10-29%, 30-49% or 50-100% reduction in personnel) and increased all-cause mortality among those remaining in the downsized workplaces. No sex differences were observed in these effects among those who remained in the downsized workplaces, nor was a period of particular vulnerability immediately following the downsizing identified. Furthermore, no detrimental effects were observed for any particular cause of death studied. CONCLUSIONS: The results provide evidence that downsizing is not a significant determinant of excess mortality among those remaining in the downsized workplaces. PMID- 18413438 TI - Delayed lupus nephritis. AB - OBJECTIVE: To describe and analyse the clinical and immunological characteristics of a large series of patients with delayed lupus nephritis (LN). METHODS: A cross sectional study was carried out. Patients with systemic lupus erythematosus (SLE) who developed renal involvement >or=5 years after the first manifestation(s) of the disease (delayed LN, n = 48) were compared with patients with SLE in whom LN developed within 5 years or less after SLE appeared (early-onset LN, n = 187). A control group, the no LN (NLN) group, comprised patients with longstanding SLE (duration of disease >10 years) who had never shown signs of renal involvement (n = 164). RESULTS: The group with delayed LN was positively associated with Sjogren's syndrome, lung involvement and antiphospholipid syndrome as compared with early LN. However, its renal clinical expression and histopathological patterns were similar to those of early-onset LN. The frequency of anti-dsDNA, anti-Sm and anti-RNP antibodies was higher in patients with LN than in the NLN group, as was the frequency of low complement levels. Jaccoud's arthropathy was a protective factor for nephritis. CONCLUSIONS: Delayed LN is not uncommon in patients with SLE. The identified risk factors might aid in its diagnosis and enhance the ability to identify patients at risk for this complication of SLE. PMID- 18413439 TI - Acute cold stress in rheumatoid arthritis inadequately activates stress responses and induces an increase of interleukin 6. AB - OBJECTIVE: Acute stress in patients with rheumatoid arthritis (RA) should stimulate a strong stress response. After cryotherapy, we expected to observe an increase of hormones of the adrenal gland and the sympathetic nervous system. METHODS: A total of 55 patients with RA were recruited for whole-body cryotherapy at -110 degrees C and -60 degrees C, and local cold therapy between -20 degrees C and -30 degrees C for 7 days. We measured plasma levels of steroid hormones, neuropeptide Y (sympathetic marker), and interleukin (IL)6 daily before and after cryotherapy. RESULTS: In both therapy groups with/without glucocorticoids (GC), hormone and IL6 levels at baseline and 5 h after cold stress did not change over 7 days of cryotherapy. In patients without GC, plasma levels of cortisol and androstenedione were highest after -110 degrees C cold stress followed by -60 degrees C or local cold stress. The opposite was found in patients under GC therapy, in whom, unexpectedly, -110 degrees C cold stress elicited the smallest responses. In patients without GC, adrenal cortisol production increased relative to other adrenal steroids, and again the opposite was seen under GC therapy with a loss of cortisol and an increase of dehydroepiandrosterone. Importantly, there was no sympathetic stress response in both groups. Patients without GC and -110 degrees C cold stress demonstrated higher plasma IL6 compared to the other treatment groups (not observed under GC), but they showed the best clinical response. CONCLUSIONS: We detected an inadequate stress response in patients with GC. It is further shown that the sympathetic stress response was inadequate in patients with/without GC. Paradoxically, plasma levels of IL6 increased under strong cold stress in patients without GC. These findings confirm dysfunctional stress axes in RA. PMID- 18413440 TI - Safety and efficacy of combination of etanercept and methotrexate compared to treatment with etanercept only in patients with juvenile idiopathic arthritis (JIA): preliminary data from the German JIA Registry. AB - OBJECTIVE: Etanercept monotherapy has been studied and approved for treatment of polyarticular juvenile idiopathic arthritis (JIA). The following study evaluates the safety and efficacy of combination therapy of etanercept and methotrexate compared to etanercept monotherapy in JIA. METHODS: We perfomed an open, non randomised study on patients who had previously failed to respond to at least one disease-modifying antirheumatic drug (DMARD). A total of 722 patients with JIA in whom at least 1 item of follow-up data was recorded were identified; of these, 118 patients treated with further slow acting drugs were excluded. In all, 504 patients were treated with a combination of etanercept and methotrexate. A total of 100 patients treated with etanercept only were in the control group. Efficacy was calculated using the American College of Rheumatology paediatric scores for 30, 50 and 70% improvement (PedACR30/50/70). Adverse events (AEs) and serious adverse events (SAEs) were reported. RESULTS: After 12 months 55 patients in the monotherapy group and 376 patients in the etanercept and methotrexate group were available for comparison. For the intention to treat analysis, 65 patients discontinuing treatment prematurely were included. All activity parameters decreased significantly in both treatment groups. After 12 months 81%/74%/62% of patients of the etanercept and methotrexate group and 70%/63%/45% of patients of the etanercept monotherapy group achieved PedACR30/50/70 scores, respectively (p<0.05 for PedACR30, p<0.01 for PedACR70). The likelihood of achieving a PedACR70 increased with combination therapy with an odds ratio of 2.1 (95% CI 1.2 to 3.5). In total, 25 infectious and 23 non-infectious SAEs including 3 malignancies occurred in the etanercept and methotrexate group, and 1 infectious and 3 non-infectious SAEs occurred in the single etanercept group. CONCLUSIONS: The patients' disease activity improved during etanercept monotherapy and etanercept and methotrexate combination therapy. Tolerability in both treatment groups was comparable. PMID- 18413441 TI - EULAR recommendations for the management of large vessel vasculitis. AB - OBJECTIVES: To develop European League Against Rheumatism (EULAR) recommendations for the management of large vessel vasculitis. METHODS: An expert group (10 rheumatologists, 3 nephrologists, 2 immunolgists, 2 internists representing 8 European countries and the USA, a clinical epidemiologist and a representative from a drug regulatory agency) identified 10 topics for a systematic literature search through a modified Delphi technique. In accordance with standardised EULAR operating procedures, recommendations were derived for the management of large vessel vasculitis. In the absence of evidence, recommendations were formulated on the basis of a consensus opinion. RESULTS: Seven recommendations were made relating to the assessment, investigation and treatment of patients with large vessel vasculitis. The strength of recommendations was restricted by the low level of evidence and EULAR standardised operating procedures. CONCLUSIONS: On the basis of evidence and expert consensus, management recommendations for large vessel vasculitis have been formulated and are commended for use in everyday clinical practice. PMID- 18413442 TI - Recurrence of spondylarthropathy among first-degree relatives of patients: a systematic cross-sectional study. AB - OBJECTIVE: To examine the recurrence of manifestations belonging to the spectrum of spondylarthropathy (SpA) in first-degree relatives of patients with SpA, and to estimate the recurrence risk ratio. METHODS: Parents and siblings of consecutive SpA probands have been thoroughly investigated, including clinical data collection, pelvic x ray and human leukocyte antigen (HLA)-B27 status determination. The diagnosis of SpA was made according to European Spondylarthropathy Study Group and/or the Amor criteria. The recurrence risk ratio lambda(1), which gives an estimate of the weight of genetic factors, was calculated as the ratio of the recurrence risk of SpA in first-degree relatives compared with the population prevalence of SpA. The lambda(non-HLA) was obtained by similar calculations restricted to HLA-B27+ individuals. RESULTS: Most manifestations of SpA were more frequent among the 157 HLA-B27+ relatives of 83 probands than among their 111 HLA-B27- relatives. A diagnosis of SpA was made in 50 relatives of 31 (37%) probands. Recurrence was very similar between parents and siblings, without gender difference, resulting in overall recurrence risk of 12% in first-degree relatives and of 22.7% in HLA-B27+ relatives. The lambda(1) value was 40 and the lambda(non-HLA) value was 6.5, very close to the lambda(HLA) value of 6.25 estimated from linkage study in SpA. CONCLUSIONS: A similar recurrence risk of SpA was observed between parents and siblings, consistent with a model of inheritance with no dominance variance and without sex influence. The weight of the non-HLA genetic component was equivalent to that estimated for the HLA locus, and fitted a model of multiplicative interaction between HLA and non HLA genetic components. PMID- 18413443 TI - Long-term follow-up on effectiveness and safety of etanercept in juvenile idiopathic arthritis: the Dutch national register. AB - OBJECTIVE: We undertook an observational study to obtain a complete overview of the long-term effectiveness and safety of etanercept in patients with different juvenile idiopathic arthritis (JIA) subtypes. METHODS: At baseline we collected patient and disease characteristics of all Dutch patients with JIA who started treatment with etanercept. Disease activity was evaluated (at start of the study, after 3 months and then yearly) according to the JIA core set of the American College of Rheumatology paediatric definition for 30, 50 and 70% improvement (ACR Pedi 30, 50 and 70). Use of etanercept and concomitant drugs was monitored. Adverse events were recorded. RESULTS: We included 146 patients with JIA with a median follow-up of 2.5 years per patient (range 0.3-7.3). JIA subtypes represented: 27% systemic, 8% polyarticular rheumatoid factor positive, 38% polyarticular rheumatoid factor negative, 19% oligoarticular extended, 3% enthesitis-related and 5% psoriatica. Most patients (77%) met the criteria of the ACR Pedi 30 in the first 3 months of treatment. For the majority of patients this improvement was sustained; 53 (36%) of all patients met the remission criteria. No other second-line agents were needed in 43 patients. Although patients with systemic JIA responded initially less to etanercept therapy than patients from other subtypes, those who did respond showed equal effectiveness in the long term. Serious adverse events rate was low (0.029 per patient year). CONCLUSIONS: Etanercept is effective and safe in JIA, even for a large proportion of the patients with systemic JIA. The greatest improvement occurred in the first 3 months of treatment, and was sustained for a long time in most patients (up to 75 months). PMID- 18413444 TI - EULAR recommendations for the management of primary small and medium vessel vasculitis. AB - OBJECTIVES: To develop European League Against Rheumatism (EULAR) recommendations for the management of small and medium vessel vasculitis. METHODS: An expert group (consisting of 10 rheumatologists, 3 nephrologists, 2 immunologists, 2 internists representing 8 European countries and the USA, a clinical epidemiologist and a representative from a drug regulatory agency) identified 10 topics for a systematic literature search using a modified Delphi technique. In accordance with standardised EULAR operating procedures, recommendations were derived for the management of small and medium vessel vasculitis. In the absence of evidence, recommendations were formulated on the basis of a consensus opinion. RESULTS: In all, 15 recommendations were made for the management of small and medium vessel vasculitis. The strength of recommendations was restricted by low quality of evidence and by EULAR standardised operating procedures. CONCLUSIONS: On the basis of evidence and expert consensus, recommendations have been made for the evaluation, investigation, treatment and monitoring of patients with small and medium vessel vasculitis for use in everyday clinical practice. PMID- 18413445 TI - Smoking increases peptidylarginine deiminase 2 enzyme expression in human lungs and increases citrullination in BAL cells. AB - OBJECTIVES: A gene-environment interaction between HLA-DR shared epitope genes and smoking in anti-cyclic citrullinated peptide antibody-positive rheumatoid arthritis (RA) has been reported. Identification of citrullinated proteins in bronchoalveolar lavage (BAL) cells from smokers has led to the suggestion that citrullination induced by smoking might be the first step in the pathogenic chain of RA. OBJECTIVE: To confirm and extend these findings. METHODS: Immunohistochemistry was performed on BAL cells and bronchial mucosal biopsy sections obtained through bronchoscopy from 14 healthy smokers and 16 healthy non smokers. Two antibodies recognising citrullinated proteins, two antibodies recognising peptidylarginine deiminase (PAD)2 enzyme and one recognising PAD4 enzyme were used. RESULTS: Citrullinated proteins are upregulated in BAL cells of healthy smokers compared with healthy non-smokers. This was associated with higher expression of the PAD2 enzyme. The same level of citrullinated proteins was present in bronchial mucosal biopsy specimens of healthy smokers and non smokers, despite higher expression of PAD2 in smokers. CONCLUSION: This study provides evidence that smoking enhances PAD2 expression in the bronchial mucosal and alveolar compartment, with consequent generation of citrullinated proteins in the latter. Smoking is an environmental factor that may lead to citrulline autoimmunity in genetically susceptible subjects. PMID- 18413446 TI - Prevention of early in life mercury exposures: no more unnecessary delays. PMID- 18413447 TI - Health reform in El Salvador: a lost opportunity for reducing health inequity and social exclusion? PMID- 18413448 TI - Widening the scope, targeting interventions, creating risk groups: maternal and child health in Denmark and Sweden from 1930s and onwards. AB - The subject of this article is the health checks for pregnant women and children in Denmark and Sweden introduced in the 1930s and 1940s. It focuses on how political goals and means have changed up until today. The questions addressed are: Which health issues did the politicians and authorities find relevant to deal with? How should they be dealt with? Who were the interventions targeted towards? It is shown that from the 1970s changes gradually took place in two respects. (1) The scope widened inasmuch as increasingly more aspects were to be included in the examinations and guidance. Not only did they come to comprise the physical, social and mental wellbeing of the children but also the life of the entire family, including efforts to improve the parents' social networks and their ability to handle their relationship with each other. (2) Interventions became increasingly targeted towards those categorised as being in need, either for medical reasons or because they were seen as less capable of caring for their children. Thus, this implied that the universal model of welfare provision was partly abandoned. The categorisations necessitated definitions of normality in more aspects and more surveillance in order to assess whether women and children lived up to the criteria of normality. These two changes were probably inter related since the urge to target and therefore identify risk groups increased when more resources were invested as a consequence of the widening scope of the activities. PMID- 18413449 TI - Is the association between childhood socioeconomic circumstances and cause specific mortality established? Update of a systematic review. AB - OBJECTIVE: To update a systematic review on the association between childhood socioeconomic circumstances and cause-specific mortality. Studies published since 2003 include a far greater number of deaths than was previously available justifying an update of the previous systematic review. METHODS: Individual-level studies examining childhood socioeconomic circumstances and adult overall and cause-specific mortality published between 2003 and April 2007. RESULTS AND CONCLUSIONS: The new studies confirmed that mortality risk for all causes was higher among those who experienced poorer socioeconomic circumstances during childhood. As already suggested in the original systematic review, not all causes of death were equally related to childhood socioeconomic circumstances. A greater proportion of new studies included women and showed that a similar pattern is valid for both genders. In addition, the new studies show that this association persists among younger birth cohorts, despite temporal general improvements in childhood conditions across successive birth cohorts. The difficulties of establishing a particular life-course model were highlighted. PMID- 18413450 TI - Enhancing global capacity in the surveillance, prevention, and control of chronic diseases: seven themes to consider and build upon. AB - BACKGROUND: Chronic diseases are now a major health problem in developing countries as well as in the developed world. Although chronic diseases cannot be communicated from person to person, their risk factors (for example, smoking, inactivity, dietary habits) are readily transferred around the world. With increasing human progress and technological advance, the pandemic of chronic diseases will become an even bigger threat to global health. METHODS: Based on our experiences and publications as well as review of the literature, we contribute ideas and working examples that might help enhance global capacity in the surveillance of chronic diseases and their prevention and control. Innovative ideas and solutions were actively sought. RESULTS: Ideas and working examples to help enhance global capacity were grouped under seven themes, concisely summarised by the acronym "SCIENCE": Strategy, Collaboration, Information, Education, Novelty, Communication and Evaluation. CONCLUSION: Building a basis for action using the seven themes articulated, especially by incorporating innovative ideas, we presented here, can help enhance global capacity in chronic disease surveillance, prevention and control. Informed initiatives can help achieve the new World Health Organization global goal of reducing chronic disease death rates by 2% annually, generate new ideas for effective interventions and ultimately bring global chronic diseases under greater control. PMID- 18413451 TI - Familial risks for chronic obstructive pulmonary disease among siblings based on hospitalisations in Sweden. AB - BACKGROUND: Chronic obstructive pulmonary disease (COPD) is a progressive disabling condition, for which tobacco smoking, environmental pollution, inherited alpha-antitrypsin deficiency and their interactions are predisposing factors. We carried out a family study on COPD in order to address the role of heritable and environmental risk factors at a population level. METHODS: In a nationwide study on familial risks for COPD the Multigeneration Register on 0-72 year-old subjects was linked to the Hospital Discharge Register from years 1987 to 2004. Standardised incidence ratios (SIRs) were calculated for affected singleton siblings, twins and spouses by comparing them with those whose siblings or spouses had no hospitalisation for COPD. RESULTS: More than 14 300 hospitalised cases and 604 affected siblings were identified. The familial SIR for obstructive chronic bronchitis was 4.65, which was higher for those diagnosed at young age but independent of sex or the age differences between the siblings. When both siblings were diagnosed with obstructive chronic bronchitis or emphysema the SIRs were 4.45 and 14.22, respectively. The SIR for obstructive chronic bronchitis in 24 twin pairs was 11.87. The SIR for spouses was about 1.6. CONCLUSIONS: The much higher risk for siblings of patients with COPD than the risk for spouses suggests that heritable effects underlie familial susceptibility to this disease. For the rare familial emphysema, alpha-antitrypsin deficiency may be an important cause. To what extent it also contributes to familial obstructive chronic bronchitis remains to be established. The anticipated gene environment interactions with sufficient sample size need to be accommodated in future aetiological studies on COPD. PMID- 18413452 TI - Do mother's education and foreign born status interact to influence birth outcomes? Clarifying the epidemiological paradox and the healthy migrant effect. AB - INTRODUCTION: The unresolved "epidemiological paradox" concerns the association between low socioeconomic status and unexpectedly favourable birth outcomes in foreign born mothers. The "healthy migrant" effect concerns the association between foreign born status per se and birth outcomes. The epidemiological paradox and healthy migrant effect were analysed for newborns in a favourable sociopolitical environment. METHODS: 98,330 live births to mothers in Montreal, Canada from 1997 to 2001 were analysed. Mothers were categorised as foreign born versus Canadian born. Outcomes were: small for gestational age (SGA) birth; low birth weight (LBW) and preterm birth (PTB). Multilevel logistic regression was used to examine the interaction between maternal education and foreign born status, adjusting for covariates. RESULTS: Not having a high school diploma was associated with LBW in Canadian (odds ratio (OR) 3.20; 95% CI 2.61 to 3.91) but not foreign born (OR 1.14; 95% CI 0.99 to 2.10) mothers and was more strongly associated with SGA birth in Canadian (OR 2.03; 95% CI 1.84 to 2.22) than in foreign born (OR 1.26; 95% CI 1.07 to 1.49) mothers. Foreign born status was associated with SGA birth (OR 1.37; 95% CI 1.28 to 1.47), LBW (OR 1.51; 95% CI 1.27 to 1.79) and PTB (OR 1.12; 95% CI 1.03 to 1.22) in university-educated mothers only. CONCLUSIONS: The epidemiological paradox associated with low educational attainment was present for SGA birth and LBW but not PTB. Foreign born status was associated with adverse birth outcomes in university-educated mothers, the opposite of the healthy migrant effect. PMID- 18413453 TI - Factors related to depression during pregnancy: are there gender differences? AB - OBJECTIVE: To determine gender differences between women and their partners in the effect of psychosocial and personal factors on depression during the third trimester of pregnancy. METHOD: A cross-sectional survey was carried out among 687 women and their partners (n = 669) attending the prenatal programme of Valencia province (Spain). Data collection was carried out by means of a self reported questionnaire. The outcome variable was depression during the third trimester of pregnancy measured by the Edinburgh Postnatal Depression Scale. Predictor variables were psychosocial (marital dissatisfaction, confidant and affective social support) and personal (previous history of depression, partner depression and unplanned pregnancy) variables. The adjusted odds ratios (aOR) and their 95% confidence intervals were calculated by fitting a logistic regression model. RESULTS: The prevalence of pregnancy depression was higher among women (10.3%) than men (6.5%). In both sexes, the probability of depression during pregnancy was higher in those with marital dissatisfaction (aOR 3.05, 95% CI 1.59 to 5.82 for women and 3.14, 95% CI 1.24 to 7.99 for men) and among those whose with a previous history of depression (aOR 2.18, 95% CI 1.22 to 3.89 for women and 5.22, 95% CI 2.05 to 13.34 for men). Unplanned pregnancy did not increase the risk of pregnancy depression either in women or men. Gender differences were found on the impact of social support and partner's depression. Whereas among men low affective social support and partner depression were associated with a higher probability of reporting depression, none of these variables were related to women's depression. CONCLUSION: Most predictor factors of depression during pregnancy are similar for both sexes but a gender-different impact of social support and partner depression on pregnancy depression was appreciated. Health professionals should be aware of potentially vulnerable groups for early diagnosis of pregnancy depression and to provide effective interventions. PMID- 18413454 TI - Effects of allostatic load on the social gradient in ischaemic heart disease and periodontal disease: evidence from the Third National Health and Nutrition Examination Survey. AB - INTRODUCTION: Psychosocial stress and allostatic load have been postulated as a mechanism explaining socioeconomic inequalities in general and oral health. This study tested whether markers of allostatic load are associated with both ischaemic heart disease and periodontal disease and whether they affect education and income gradients for both conditions. METHODS: Data are from the Third National Health and Nutrition Examination Survey, conducted in the United States from 1988 to 1994. Ischaemic heart disease was determined by the presence of angina or diagnosis of heart attack. Four variables were used for periodontal disease. Individual and aggregate markers of allostatic load were used. RESULTS: Allostatic load (both aggregate and most individual markers) was associated with higher probabilities of all examined health outcomes. Adjusting for markers of allostatic load attenuated education and income gradients in both ischaemic heart disease and periodontal disease. The relationship between socioeconomic position and the examined health outcomes remained significant. CONCLUSION: Indicators of allostatic load were associated with ischaemic heart disease and periodontal disease and had a mediating effect partly explaining the social gradients in both diseases. The results suggest a possible common stress pathway linking socioeconomic position to both conditions. PMID- 18413455 TI - Prospective association between physical activity and falls in community-dwelling older women. AB - OBJECTIVE: To explore associations between physical activity and the risk of falls and fractured bones in community-dwelling older women. DESIGN, SETTING AND PARTICIPANTS: A prospective observational survey with three and six-year follow ups. The sample included 8188 healthy, community-dwelling women, aged 70-75 years in 1996, who completed surveys as participants in the Australian Longitudinal Study on Women's Health. Women who reported a recent serious injury from falling were excluded. Outcomes were reports of a fall to the ground, injury from a fall, and a fractured bone in 1999 and 2002. The main predictor variable was physical activity level in 1996, categorised on the basis of weekly frequency as none/very low, low, moderate, high and very high. Covariates were demographic and health related variables. Logistic regression models were computed separately for each outcome in 1999 and 2002. MAIN RESULTS: In multivariable models, very high physical activity was associated with a decreased risk of reporting a fall in 1999 (odds ratio (OR) 0.67; 95% CI 0.47 to 0.95) and in 2002 (OR 0.64; 95% CI 0.43 to 0.96). High/very high physical activity was associated with a decreased risk of a fractured bone in 2002 (OR 0.53; 95% CI 0.34 to 0.83). No significant association was found between physical activity and injury from a fall. CONCLUSIONS: The results suggest that at least daily moderate to vigorous intensity physical activity is required for the primary prevention of falls to the ground and fractured bones in women aged 70-75 years. PMID- 18413456 TI - Patient expectations are not always the same. AB - OBJECTIVE: To validate a scale that measures patients' expectations when seeking advice for health problems of different types. METHODS: 360 patients who had consulted their general practitioner (GP) during the previous 12 months were randomly selected from the lists of 30 GPs. A questionnaire, including a 13-item expectation scale, was administered by interview in the patient's home to assess expectations in relation to five health problems, three biomedical (strong chest pain, genital discharge and the common cold) and two psychosocial (depression/sadness and serious family problem), repeating the expectation scale for each one. The frequency distribution of items was analysed, multi-level factorial analysis was performed and the reliability of the expectation scale was tested for each hypothetical clinical condition. RESULTS: The response rate was 90%. Mean age of patients was 47.3 years (SD 16.5); 51% were women. Expectations were high but varied according to the nature and severity of the condition. The percentage of patients wanting the doctor alone to make decisions ranged from 50% for "family problem" to 68% for "chest pain". The five factorial structures differed and explained 49.3-63.9% of the variance. Similarities were observed depending on the type of problem. "Communication" and "Experience of disease" were thus separate dimensions for the biomedical diseases but mixed for the psychosocial conditions. CONCLUSIONS: The factorial structure of expectations varied, indicating that expectations are not homogeneous in all clinical situations. The desire of the patient to participate in decision-making also differs according to the type of health problem. PMID- 18413457 TI - Cultural identity, clothing and common mental disorder: a prospective school based study of white British and Bangladeshi adolescents. AB - OBJECTIVE: Cultural integration is the healthiest outcome for young people living in multicultural societies. This paper investigates the influence of different cultural identities on the risk of common mental disorders among Bangladeshi and white British pupils. DESIGN: The cultural identity of 11-14-year-old school pupils was assessed by their preferences for friends and clothes of their own or other cultural groups; using this information pupils were classified into traditional, integrated, assimilated or marginalised groups. We undertook prospective analyses of cultural identity and its impacts on the later mental health of young people. SETTING: East London. PARTICIPANTS: In 2001, white British (573) and Bangladeshi (682) school pupils from a representative sample of schools completed a self-report questionnaire that assessed their cultural, social and health characteristics. In 2003, 383 white British and 517 Bangladeshi pupils were resurveyed and completed measures of mental health. MAIN OUTCOME MEASURE: Strengths and difficulties questionnaire. RESULTS: Bangladeshi pupils preferring clothes from their own cultural group (traditional clothing) were less likely to have later mental health problems when compared with Bangladeshi pupils showing an equal preference for clothing from their own and other cultures (integrated clothing; odds ratio (OR) 0.3, 95% CI 0.1 to 0.9). In gender-specific analyses, this finding was sustained only among Bangladeshi girls (OR 0.1, 95% CI 0.1 to 0.7). Integrated clothing choices were least risky only for white British adolescents. Friendship choices showed no prospective associations with later mental health problems. CONCLUSIONS: Cultural identity, expressed by clothing preferences, influences mental health; the effects differ by gender and ethnic group. PMID- 18413458 TI - Atmospheric pollutants and mortalities in English local authority areas. AB - OBJECTIVES: To measure geographical co-relationships between disease-specific standardised mortality ratios (SMR) and different atmospheric emissions in 352 English local authorities. To link specific exposures with specific causes of death and to identify responsible polluting sources. To see whether long-term moderate exposures have the same lethal effects as short-term high-pollution (ie, smog) episodes. DESIGN: Geographical distributions of SMR, atmospheric emissions and social hazards, extracted from three different sources, were converted to a congruent format. Correlation coefficients were calculated within and between these different datasets. Mortality/pollutant correlations were recalculated after additionally standardising the SMR for social differences between local authorities. SETTING: The 352 English local authority areas, 1996-2004. MAIN RESULTS: SMR for one group of diseases (including upper alimentary and respiratory cancers, ischaemic heart disease, peptic ulcer, pneumonia) were related to a range of combustion emissions and to multiple social deprivation, cigarette smoking, binge drinking and a northern location. Additional standardisation of all SMR for these social hazards left a small subgroup independently related to atmospheric pollution, mainly from oil combustion. Correlations with pneumonia deaths were exceptional. CONCLUSIONS: High mortality rates were observed in areas with elevated ambient pollution levels. The strongest single effect was an increase in pneumonia deaths. Road transport was the chief source of the emissions responsible, although it was not possible to discriminate between the different chemical components. Many "pneumonia" deaths were probably caused by direct chemical injury, as in the 1952 London smog and are better regarded as "acute respiratory distress syndrome" or "acute lung injury". PMID- 18413459 TI - Increased use of antidepressants and decreasing suicide rates: a population-based study using Danish register data. AB - OBJECTIVE: The objective of the present study was to examine if the change in the suicide rate is associated with individuals' use of antidepressants as has been suggested by ecological studies. DESIGN: Decomposition of suicide rates by antidepressant treatment group. SETTING: Population-based record linkage. PARTICIPANTS: All individuals aged 50 years and older living in Denmark between 1 January 1996 and 31 December 2000 (N = 2,100,808). MAIN OUTCOME MEASURES: Suicide rates are calculated according to current antidepressant treatment status (no treatment, tricyclic antidepressants (TCA), selective serotonin reuptake inhibitors (SSRI), other antidepressants). The change in the suicide rate during 1996-2000 was decomposed by treatment group. RESULTS: Only one in five older adults dying by suicide was in treatment at the time of death. Whereas the male suicide rate declined by 9.7 suicides per 100,000, recipients of antidepressants contributed to the decline by 0.9 suicides. Women redeeming antidepressant prescriptions accounted for 0.4 suicides of the observed reduction of 3.3 per 100,000. The average suicide rates for men receiving TCA and SSRI were 153.3 and 169.0 per 100,000 person-years, respectively. Among older women, both TCA and SSRI users had an average suicide rate of 68.8 per 100,000 over the period examined. CONCLUSIONS: Just a small proportion of older adults dying by suicide were found to be in treatment with antidepressants at the time of death. Individuals in active treatment with antidepressants seem to account for 10% of the decline in the suicide rate. Nevertheless, suicides might be prevented by more effective treatment. PMID- 18413460 TI - Predictors of symptoms of post-traumatic stress disorder after the AZF chemical factory explosion on 21 September 2001, in Toulouse, France. AB - OBJECTIVE: To analyse in the general population the prevalence and predictors of symptomatology consistent with post-traumatic stress disorder (S-PTSD) 18 months after an industrial explosion. DESIGN: Cross-sectional survey. PARTICIPANTS AND OUTCOME MEASURES: A random sample of 1191 city inhabitants, including an oversample of the immediate area (<3 km). S-PTSD was measured by the self administered Impact of Event Scale-Revised. The relation between S-PTSD and individual vulnerability factors, immediate exposure and post-trauma factors was analysed by gender. RESULTS: S-PTSD was more prevalent in the immediate area than in the peripheral area (women 19% vs 8%; men 8% vs 2%, p<0.01). In the immediate area, S-PTSD was independently associated with birth outside France (men: OR(a) = 13.9, 95% CI 3.7 to 52.8; women: OR(a) = 2.1, 95% CI 1.0 to 4.2), age more than 40 years (men: OR(a) = 4.3, 95% CI 1.01 to 18.2; women: OR(a) = 2.3, 95% CI 1.1 to 4.5), previous psychotropic treatment (men: OR(a) = 11.5, 95% CI 2.4 to 53.6), proximity to the explosion (less educated men only) (OR(a) = 9.3, 95% CI 1.9 to 44.7), rescue efforts (men: OR(a) = 5.2, 95% CI 1.5 to 18.2), temporarily uninhabitable home (men: OR(a) = 5.8, 95% CI 1.9 to 18.1), personal injury (women: OR(a) = 3.7, 95% CI 1.7 to 8.4), financial difficulties (men: OR(a) = 17.4, 95% CI 4.2 to 72.1; women: OR(a) = 3.4, 95% CI 1.7 to 7.1) and inconvenience due to closure of public services (women: OR(a) = 4.1, 95% CI 1.6 to 9.9). CONCLUSIONS: Individual vulnerability, exposure and post-trauma factors were associated with S-PTSD. Vulnerable subgroups, defined by low socioeconomic characteristics may warrant focused screening after such disasters. PMID- 18413461 TI - Impact of parity on anthropometric measures of obesity controlling by multiple confounders: a cross-sectional study in Chilean women. AB - AIM: To find out whether there is an association between parity and obesity, evaluated through body mass index (BMI), waist circumference (WC), waist-to-hip ratio (WHR) and waist-to-height ratio (WHtR) in Chilean women after controlling for sociodemographic characteristics, health risk and gynaeco-obstetric factors. DESIGN: Cross-sectional study, using baseline data of the San Francisco Project. SETTING: San Francisco de Mostazal, located in the central region of Chile, 6512 Chilean-Hispanic women (Spanish heritage with a variable indigenous component). METHODS: A weighted random sample of 508 women who had their first pregnancy inside the primary child-bearing ages. Data were collected between 1997 and 1999. Statistical associations between parity and different anthropometric measurements of adiposity in multiple linear (MLnR) and logistic regression models (MLtR) were evaluated. RESULTS: In MLnR a modest parity-related increment in BMI and practically null increment in WC, WHR and WHtR was observed. Covariates that showed a statistically significant association with anthropometric measures of adiposity were age, low education, marital status, employment, smoking, smoking cessation, hypertension, diabetes, dyslipidaemia, parent's obesity, menarche and fetal macrosomia. Crude odds ratio (OR) showed a strong association between parity and anthropometric markers of obesity. Nevertheless, after adjustments in MLtR models, the association remained only for BMI. All the measures of abdominal obesity related to parous women showed OR smaller than 1 (95% confidence intervals 0.57 to 0.96). CONCLUSIONS: Parity modestly influences BMI, but does not seem to be related to WC, WHR and WHtR after controlling by confounders. Parity can increase adiposity but not necessarily following an abdominal pattern. PMID- 18413465 TI - How to predict the risk of Parkinson disease in relatives of parkin mutation carriers: a complex puzzle of age, penetrance, and number of mutated alleles. PMID- 18413466 TI - The changing face of neural stem cell therapy in neurologic diseases. AB - New insights into the biology of neural stem cells (NSCs) have raised expectations for their use in the treatment of neurologic diseases. Originally, NSC transplantation was proposed as a means of replacing cells in central nervous system diseases that result in cell loss. However, recent data regarding their beneficial effects in various animal models of neurologic diseases indicate that transplanted NSCs may also attenuate deleterious inflammation, protect the central nervous system from degeneration, and enhance endogenous recovery processes. Herein, we review recent developments and future prospects of NSC therapy in neurologic diseases. PMID- 18413464 TI - Serum urate as a predictor of clinical and radiographic progression in Parkinson disease. AB - OBJECTIVE: To determine whether concentration of serum urate, a purine metabolite and potent antioxidant that has been linked to a reduced risk of Parkinson disease (PD), predicts prognosis in PD. DESIGN: Prospective study. SETTING: The Parkinson Research Examination of CEP-1347 Trial (PRECEPT) study, which investigated the effects of a potential neuroprotectant on rates of PD progression, was conducted between April 2002 and August 2005 (average follow-up time 21.4 months). PARTICIPANTS: Eight hundred four subjects with early PD enrolled in the PRECEPT study. MAIN OUTCOME MEASURES: The primary study end point was progression to clinical disability sufficient to warrant dopaminergic therapy. Cox proportional hazards models were used to estimate the hazard ratio (HR) of reaching end point according to quintiles of baseline serum urate concentration, adjusting for sex, age, and other potential covariates. Change in striatal uptake of iodine I 123-labeled 2-beta-carbomethoxy-3-beta-(4 iodophenyl)tropane ([(123)I]beta-CIT), a marker for the presynaptic dopamine transporter, was assessed with linear regression for a subset of 399 subjects. RESULTS: The adjusted HR of reaching end point declined with increasing baseline concentrations of urate; subjects in the top quintile reached the end point at only half the rate of subjects in the bottom quintile (HR, 0.51; 95% confidence interval [CI], 0.37-0.72; P for trend < .001). This association was markedly stronger in men (HR, 0.39; 95% CI, 0.26-0.60; P for trend < .001) than in women (HR, 0.77; 95% CI, 0.39-1.50; P for trend = .33). The percentage of loss in striatal [(123)I]beta-CIT uptake also improved with increasing serum urate concentrations (overall P for trend = .002; men, P = .001; women, P = .43). CONCLUSIONS: These findings identify serum urate as the first molecular factor directly linked to the progression of typical PD and suggest that targeting urate or its determinants could be an effective disease-modifying therapy in PD. Trial Registration clinicaltrials.gov Identifier: NCT00040404. PMID- 18413467 TI - Refining frontotemporal dementia with parkinsonism linked to chromosome 17: introducing FTDP-17 (MAPT) and FTDP-17 (PGRN). AB - Frontotemporal dementia with parkinsonism (FTDP) is a major neurodegenerative syndrome, particularly for those with symptoms beginning before age 65 years. A spectrum of degenerative disorders can present as sporadic or familial FTDP. Mutations in the gene encoding the microtubule-associated protein tau (MAPT; OMIM +157140) on chromosome 17 have been found in many kindreds with familial FTDP. Several other kindreds with FTDP had been linked to chromosome 17, but they had ubiquitin-positive inclusions rather than tauopathy pathology and no mutations in MAPT. This conundrum was solved in 2006 with the identification of mutations in the gene encoding progranulin (PGRN; OMIM *138945), which is only 1.7 Mb centromeric to MAPT on chromosome 17. In this review, we compare and contrast the demographic, clinical, radiologic, neuropathologic, genetic, and pathophysiologic features in patients with FTDP linked to mutations in MAPT and PGRN, highlighting the many similarities but also a few important differences. Our findings describe an intriguing oddity of nature in which 2 genes can cause a similar phenotype through apparently different mechanisms yet reside so near to each other on the same chromosome. PMID- 18413469 TI - Glucose homeostasis in Huntington disease: abnormalities in insulin sensitivity and early-phase insulin secretion. AB - BACKGROUND: Patients with Huntington disease (HD) develop diabetes mellitus more often than do matched healthy controls. Recent studies in neurodegenerative diseases suggested that insulin resistance constitutes a metabolic stressor that interacts with a preexisting neurobiological template to induce a given disorder. OBJECTIVE: To investigate possible changes in insulin sensitivity and secretion, major determinants of glucose homeostasis, in a group of consecutive normoglycemic patients with HD. DESIGN: Metabolic investigations. PARTICIPANTS: Twenty-nine untreated, nondiabetic patients with HD and 22 control participants matched by age, sex, and socioeconomic background. MAIN OUTCOME MEASURES: Glucose tolerance, assessed by means of the glucose curve during oral glucose challenge; insulin sensitivity, assessed using homeostasis model assessment and minimal model analysis based on frequent sampling of plasma glucose and plasma insulin during the intravenous glucose tolerance test; and insulin secretion, determined by means of the acute insulin response and the insulinogenic index. RESULTS: The evaluation of insulin sensitivity using homeostasis model assessment demonstrated higher homeostasis model assessment insulin resistance indices, and a lower sensitivity index when the minimal model approach was used, in patients with HD compared with controls (P = .03 and P = .003, respectively). In the assessment of early-phase insulin secretion, the acute insulin response and the insulinogenic index were lower in patients with HD compared with controls (P = .02). The number of CAG repeats correlated significantly only with acute insulin response (P = .003). CONCLUSIONS: Besides impairment in insulin secretion capacity, a simultaneous decrease in insulin sensitivity, with an increase in the insulin resistance level, was found in normoglycemic patients with HD compared with controls. These data imply that progression of the insulin secretion defect in HD may lead to a failure to compensate for insulin resistance. PMID- 18413470 TI - Rapid eye movement sleep disturbances in Huntington disease. AB - BACKGROUND: Sleep disorders including insomnia, movements during sleep, and daytime sleepiness are common but poorly studied in Huntington disease (HD). OBJECTIVE: To evaluate the HD sleep-wake phenotype (including abnormal motor activity during sleep) in patients with various HD stages and the length of CAG repeats. Because a mild hypocretin deficiency has been found in the brains of some patients with HD (hereinafter referred to as HD patients), we also tested the HD patients for narcolepsy. DESIGN AND PATIENTS: Twenty-five HD patients (including 2 premanifest carriers) underwent clinical interview, nighttime video and sleep monitoring, and daytime multiple sleep latency tests. Their results were compared with those of patients with narcolepsy and control patients. RESULTS: The HD patients had frequent insomnia, earlier sleep onset, lower sleep efficiency, increased stage 1 sleep, delayed and shortened rapid eye movement (REM) sleep, and increased periodic leg movements. Three HD patients (12%) had REM sleep behavior disorders. No sleep abnormality correlated with CAG repeat length. Reduced REM sleep duration (but not REM sleep behavior disorders) was present in premanifest carriers and patients with very mild HD and worsened with disease severity. In contrast to narcoleptic patients, HD patients had no cataplexy, hypnagogic hallucinations, or sleep paralysis. Four HD patients had abnormally low (< 8 minutes) daytime sleep latencies, but none had multiple sleep onset REM periods. CONCLUSIONS: The sleep phenotype of HD includes insomnia, advanced sleep phase, periodic leg movements, REM sleep behavior disorders, and reduced REM sleep but not narcolepsy. Reduced REM sleep may precede chorea. Mutant huntingtin may exert an effect on REM sleep and motor control during sleep. PMID- 18413468 TI - Risk of Parkinson disease in carriers of parkin mutations: estimation using the kin-cohort method. AB - OBJECTIVE: To estimate the risk of Parkinson disease (PD) in individuals with mutations in the Parkin gene. DESIGN: We assessed point mutations and exon deletions and duplications in the Parkin gene in 247 probands with PD (age at onset < or =50 years) and 104 control probands enrolled in the Genetic Epidemiology of Parkinson's Disease (GEPD) study. For each first-degree relative, a consensus diagnosis of PD was established. The probability that each relative carried a mutation was estimated from the proband's Parkin carrier status using Mendelian principles and from the relationship of the relative to the proband. SETTING: Tertiary care movement disorders center. Patients Cases, controls, and their first-degree relatives were enrolled in the GEPD study. MAIN OUTCOME MEASURES: Estimated age-specific penetrance in first-degree relatives. RESULTS: Parkin mutations were identified in 25 probands with PD (10.1%), 18 (72.0%) of whom were heterozygotes. One Parkin homozygote was reported in 2 siblings with PD. The cumulative incidence of PD to age 65 years in carrier relatives (age specific penetrance) was estimated to be 7.0% (95% confidence interval, 0.4% 71.9%), compared with 1.7% (95% confidence interval, 0.8%-3.4%) in noncarrier relatives of the cases (P = .59) and 1.1% (95% confidence interval, 0.3%-3.4%) in relatives of the controls (compared with noncarrier relatives, P = .52). CONCLUSIONS: The cumulative risk of PD to age 65 years in a noncarrier relative of a case with an age at onset of 50 years or younger is not significantly greater than the general population risk among controls. Age-specific penetrance among Parkin carriers, in particular heterozygotes, deserves further study. PMID- 18413471 TI - Cryptogenic epileptic syndromes related to SCN1A: twelve novel mutations identified. AB - BACKGROUND: Sodium channel alpha 1 subunit gene, SCN1A, is the gene encoding the neuronal voltage-gated sodium channel alpha 1 subunit (Na(v)1.1) and is mutated in different forms of epilepsy. Mutations in this gene were observed in more than 70% of patients with severe myoclonic epilepsy of infancy (SMEI) and were also found in different types of infantile epileptic encephalopathy. OBJECTIVE: To search for disease-causing mutations in SCN1A in patients with cryptogenic epileptic syndromes (ie, syndromes with an unknown cause). DESIGN: Clinical characterization and molecular genetic analysis of a cohort of patients. SETTING: University hospitals, rehabilitation centers, and molecular biology laboratories. PATIENTS: Sixty unrelated patients with cryptogenic epileptic syndromes. MAIN OUTCOME MEASURES: Samples of DNA were analyzed for mutations and for large heterozygous deletions encompassing the SCN1A gene. A search for microdeletions in the SCN1A gene was also performed in the subset of patients with SMEI/SMEI borderland who had negative results at the point mutation screening. RESULTS: No large deletions at the SCN1A locus were found in any of the patients analyzed. In contrast, 13 different point mutations were identified in 12 patients: 10 with SMEI, 1 with generalized epilepsy with febrile seizures plus, and 1 with cryptogenic focal epilepsy. An additional search for SCN1A intragenic microdeletions in the remaining patients with SMEI/SMEI-borderland and no point mutations was also negative. CONCLUSIONS: These results confirm the role of the SCN1A gene in different types of epilepsy, including cryptogenic epileptic syndromes. However, large deletions encompassing SCN1A were not common disease causing rearrangements in this group of epilepsies. PMID- 18413472 TI - Abnormal nerve conduction features in fragile X premutation carriers. AB - BACKGROUND: Distal neuropathy is part of the clinical phenotype in most males with the fragile X-associated tremor/ataxia syndrome (FXTAS) caused by the 55 to 200 CGG repeat expansion. METHODS: We performed nerve conduction studies in 16 male carriers with FXTAS, 11 non-FXTAS carriers, and 11 control subjects and assessed the outcomes with respect to the fragile X mental retardation 1 genotype (FMR1) (Online Mendelian Inheritance in Man [OMIM] 309550; NT011681) and messenger RNA expression. RESULTS: Men with FXTAS had slower tibial nerve conduction velocities and prolonged F-wave latencies compared with controls (z = 2.06, P = .04; and z = 2.73, P = .005) and unaffected premutation males (z = 1.98, P = .04; and z = 2.00, P = .04). Compound muscle action potential amplitudes were smaller in the FXTAS group relative to controls. Sural nerve action potential amplitudes were reduced in the FXTAS group compared with controls. After controlling for age, there was a significant relationship between the longer CGG repeat number and tibial nerve conduction velocity slowing (r = 0.42, P = .04) and between elevated messenger RNA levels and reduction of the tibial compound muscle action potential velocity (r = -0.52, P = .01) in the permutation group. CONCLUSIONS: Male premutation carriers had significant conduction abnormalities of motor and sensory nerves that correlated with molecular measures, suggesting that the premutation FMR1 genotype is a causal factor. There was also evidence of nerve conduction abnormalities in non-FXTAS carriers compared with controls, which suggests that the neuropathy can occur without the full clinical presentation of FXTAS. PMID- 18413473 TI - Clinical and neuropathological features of the arctic APP gene mutation causing early-onset Alzheimer disease. AB - BACKGROUND: A majority of mutations within the beta-amyloid region of the amyloid precursor protein (APP) gene cause inherited forms of intracerebral hemorrhage. Most of these mutations may also cause cognitive impairment, but the Arctic APP mutation is the only known intra-beta-amyloid mutation to date causing the more typical clinical picture of Alzheimer disease. OBJECTIVE: To describe features of 1 Swedish and 1 American family with the previously reported Arctic APP mutation. DESIGN, SETTING, AND PARTICIPANTS: Affected and nonaffected carriers of the Arctic APP mutation from the Swedish and American families were investigated clinically. In addition, 1 brain from each family was investigated neuropathologically. RESULTS: The clinical picture, with age at disease onset in the sixth to seventh decade of life and dysfunction in multiple cognitive areas, is indicative of Alzheimer disease and similar to the phenotype for other Alzheimer disease APP mutations. Several affected mutation carriers displayed general brain atrophy and reduced blood flow of the parietal lobe as demonstrated by magnetic resonance imaging and single-photon emission computed tomography. One Swedish case and 1 American case with the Arctic APP mutation came to autopsy, and both showed no signs of hemorrhage but revealed severe congophilic angiopathy, region-specific neurofibrillary tangle pathological findings, and abundant amyloid plaques. Intriguingly, most plaques from both of these cases had a characteristic ringlike character. CONCLUSIONS: Overall, our findings corroborate that the Arctic APP mutation causes a clinical and neuropathological picture compatible with Alzheimer disease. PMID- 18413474 TI - Parietal lobe deficits in frontotemporal lobar degeneration caused by a mutation in the progranulin gene. AB - OBJECTIVE: To describe the clinical, neuropsychologic, and radiologic features of a family with a C31LfsX35 mutation in the progranulin gene CCDS11483.1). DESIGN: Case series. PATIENTS: A large British kindred (DRC255) with a PGRN mutation was assessed. Affected individuals presented with a mean age of 57.8 years (range, 54 67 years) and a mean disease duration of 6.1 years (range, 2-11 years). RESULTS: All patients exhibited a clinical and radiologic phenotype compatible with frontotemporal lobar degeneration based on current consensus criteria. However, unlike sporadic frontotemporal lobar degeneration, parietal deficits, consisting of dyscalculia, visuoperceptual /visuospatial dysfunction, and/or limb apraxia, were a common feature, and brain imaging showed posterior extension of frontotemporal atrophy to involve the parietal lobes. Other common clinical features included language output impairment with either dynamic aphasia or nonfluent aphasia and a behavioral syndrome dominated by apathy. CONCLUSION: We suggest that parietal deficits may be a prominent feature of PGRN mutations and that these deficits may be caused by disruption of frontoparietal functional pathways. PMID- 18413475 TI - Patients homozygous and heterozygous for SNCA duplication in a family with parkinsonism and dementia. AB - BACKGROUND: Multiplication of the alpha-synuclein gene (SNCA) (OMIM 163890) has been identified as a causative mutation in hereditary Parkinson disease or dementia with Lewy bodies. OBJECTIVE: To determine the genetic, biochemical, and neuropathologic characteristics of patients with autopsy-confirmed autosomal dominant Lewy body disease, with particular reference to the dosage effects of SNCA. DESIGN: Four-generation family study. SETTING: Academic research. Patients We fractionated samples extracted from frozen brain tissues of 4 patients for biochemical characterization, followed by immunoblot analysis. MAIN OUTCOME MEASURES: We determined the dosages of SNCA and its surrounding genes by quantitative polymerase chain reaction analysis. RESULTS: Quantitative polymerase chain reaction analysis revealed that 3 patients were heterozygous for SNCA duplication and 1 patient was homozygous for SNCA duplication. The homozygous patient showed earlier age at onset and earlier death, with more severe cognitive impairment than the heterozygous patients. Biochemical analysis revealed that phosphorylated alpha-synuclein accumulated in the sarkosyl-insoluble urea extracted fraction of the brains of the patients. CONCLUSIONS: Pathologically confirmed Lewy body disease clinically characterized by progressive parkinsonism and cognitive dysfunction is caused by SNCA duplication. The homozygous patient demonstrated the most severe phenotype, suggesting that SNCA dosage has a considerable effect on disease phenotype even within a family. SNCA duplication results in the hyperaccumulation of phosphorylated alpha-synuclein in the brains of patients. PMID- 18413476 TI - Lack of spartin protein in Troyer syndrome: a loss-of-function disease mechanism? AB - BACKGROUND: Hereditary spastic paraplegias (SPG1-SPG33) are characterized by progressive spastic weakness of the lower limbs. A nucleotide deletion (1110delA) in the (SPG20; OMIM 275900) spartin gene is the origin of autosomal recessive Troyer syndrome. This mutation is predicted to cause premature termination of the spartin protein. However, it remains unknown whether this truncated spartin protein is absent or is present and partially functional in patients. OBJECTIVE: To determine whether the truncated spartin protein is present or absent in cells derived from patients with Troyer syndrome. DESIGN: Case report. SETTING: Academic research. PATIENTS: We describe a new family with Troyer syndrome due to the 1110delA mutation. MAIN OUTCOME MEASURES: We cultured primary fibroblasts and generated lymphoblasts from affected individuals, carriers, and control subjects and subjected these cells to immunoblot analyses. RESULTS: Spartin protein is undetectable in several cell lines derived from patients with Troyer syndrome. CONCLUSIONS: Our data suggest that Troyer syndrome results from complete loss of spartin protein rather than from the predicted partly functional fragment. This may reflect increased protein degradation or impaired translation. PMID- 18413477 TI - Muscle excitability abnormalities in Machado-Joseph disease. AB - OBJECTIVES: To estimate the frequency of and to characterize muscle excitability abnormalities in Machado-Joseph disease (MJD). DESIGN: Machado-Joseph disease is a common autosomal dominant cerebellar ataxia caused by an unstable CAG trinucleotide repeat expansion. Muscle cramps and fasciculations are frequent and sometimes disabling manifestations. However, their frequency and pathophysiological mechanisms remain largely unknown. Symptomatic patients with MJD (hereinafter MJD patients) with molecular confirmation were assessed prospectively. A standard questionnaire addressing clinical features of muscle cramps and fasciculations was used. The Cramps Disability Scale was used to quantify cramps-related disability. Patients underwent neurophysiological testing with routine techniques. F waves of the right median nerves were obtained, and persistence indexes were calculated. Four muscles (deltoid, first dorsal interossei, tibialis anterior, and vastus lateralis) were examined by needle electromyography. A semiquantitative scale (from 0 [no activity] to 4 [continuous activity]) was used to determine the frequency of rest fasciculations in each muscle. RESULTS: Fifty MJD patients (29 men) were included in the study. Their mean age at examination was 46.3 years, their mean age at onset of the disease was 35 years, and the mean duration of disease was 11.2 years. Abnormal CAG(n) varied from 59 to 75 repeats. Forty-one patients presented with muscle cramps; in 10, this was their first symptom. The frequency of cramps varied between 1 and 90 episodes a week. For 15 patients, cramps were the chief complaint, frequently disturbing sleep or work (Cramps Disability Scale score, 2 or 3). Lower limbs were affected in 37 individuals, but unusual regions, such as the face and abdominal muscles, were also involved. Fasciculations were found in 25 individuals; in 8 patients, they included facial muscles. However, fasciculations were not a significant complaint for any of these patients. The clinical and neurophysiological profile of MJD patients with and without cramps was not significantly different. However, MJD patients with fasciculations had more severe damage to their peripheral nerves. CONCLUSIONS: Muscle excitability abnormalities were found in 41 MJD patients (82%), and they were the presenting complaint in 10 (20%). They are related to altered excitability of peripheral motor axons, but mechanisms underlying cramps and fasciculations are possibly distinct in MJD patients. PMID- 18413478 TI - Impaired eye movements in presymptomatic spinocerebellar ataxia type 6. AB - BACKGROUND: Early detection of impaired neurological function in neurodegenerative diseases may aid in understanding disease pathogenesis and timing of therapeutic trials. OBJECTIVE: To identify early abnormalities of ocular motor function in individuals who have the spinocerebellar ataxia type 6 (SCA6) gene (CACNA1A) but no clinical symptoms. DESIGN: Physiological techniques were used to record and analyze eye movements and postural sway. PATIENTS: Four presymptomatic and 5 ataxic patients with SCA6, genetically identified, and 10 healthy controls. RESULTS: Presymptomatic individuals had normal postural sway but definite ocular motor abnormalities. Two had a low-amplitude horizontal gaze evoked nystagmus, 1 of whom had a significantly decreased eye velocity for upward saccades and an abnormal frequency of square-wave jerks. Another had abnormal square-wave jerks and a fourth had a reduced gain for pursuit tracking. Not all of the presymptomatic patients had the same findings, but a multivariate analysis discriminated the presymptomatic patients, as a group, from healthy controls and the ataxic patients. CONCLUSIONS: Among the earliest functional deficits in SCA6 are eye movement abnormalities, including impaired saccade velocity, saccade metrics, and pursuit gain. This suggests that early functional impairments are caused by cellular dysfunction and/or loss in the posterior cerebellar vermis and flocculus. These findings might help to determine the timing of a treatment and to define variables that could be used as outcome measures for the efficacy of therapeutic trials. PMID- 18413479 TI - Geriatric neurogenetics: oxymoron or reality? AB - BACKGROUND: Primary genetic diseases are generally associated with pediatric and young adult populations. Little information is available about the occurrence of single-gene mendelian diseases in elderly populations. OBJECTIVE: To describe the occurrence of single-gene neurogenetic disorders in a group of elderly patients. DESIGN: Retrospective review of neurogenetic cases in an academic medical center. SETTING: Academic university and Veterans Affairs medical centers. PATIENTS: Eight elderly patients with single-gene neurogenetic diseases were studied. These patients included an 87-year-old man and an 85-year-old man with Huntington disease, an 84-year-old woman with limb-girdle muscular dystrophy type 2A, a 78 year-old man with spinocerebellar ataxia type 14, an 86-year-old man with spinocerebellar ataxia type 5, an 85-year-old man with a presenilin 1 familial Alzheimer disease mutation, an 87-year-old man with autosomal dominant hereditary neuropathy, and a 78-year-old man with spinocerebellar ataxia type 6. Three patients had no family history of neurologic disease. MAIN OUTCOME MEASURES: Medical histories, physical examination results, and genetic testing results. CONCLUSIONS: Single-gene mendelian neurogenetic diseases can be found in the oldest old population (> 85 years). Such cases are currently underrecognized and will become more commonly observed in the future. This phenomenon is a result of (1) the aging of the general population, (2) better recognition of the highly variable ages at onset of genetic diseases, and (3) the availability of specific DNA-based genetic testing. PMID- 18413480 TI - Carbon 11-labeled pittsburgh compound B positron emission tomographic amyloid imaging in patients with APP locus duplication. AB - OBJECTIVE: To investigate amyloid accumulation by carbon 11-labeled Pittsburgh Compound B (11C-PiB) in hereditary cerebral amyloid angiopathy and APP locus duplication. DESIGN, SETTING, AND PATIENTS: Positron emission tomography with 11C PiB and magnetic resonance imaging were performed for 2 patients, 49-year-old and 60-year-old siblings with APP locus duplication, with hereditary Alzheimer disease and cerebral amyloid angiopathy. MAIN OUTCOME MEASURE: Change in 11C-PiB uptake. RESULTS: Uptake of 11C-PiB was increased especially in the striatum (caudate nucleus to 225% and 280% of the control mean and putamen to 166% and 185% of the control mean) and in the posterior cingulate (to 168% and 198% of the control mean), and it was marginally increased in other cortical brain areas. The pattern of increased 11C-PiB uptake was different from that seen in sporadic Alzheimer disease. CONCLUSIONS: Amyloid imaging with 11C-PiB positron emission tomography is a useful tool for detecting in vivo amyloid accumulation in patients with hereditary cerebral amyloid angiopathy. However, the pattern of 11C PiB accumulation differs between patients with typical AD and patients with APP locus duplication. PMID- 18413481 TI - In vivo detection of thalamic gliosis: a pathoradiologic demonstration in familial fatal insomnia. AB - BACKGROUND: Increasing evidence supports the usefulness of brain magnetic resonance imaging (MRI) for the diagnosis of human prion diseases. From the neuroradiological point of view, fatal familial insomnia is probably the most challenging to diagnose because brain lesions are mostly confined to the thalamus. OBJECTIVE: To determine whether multisequence MRI of the brain can show thalamic alterations and establish pathoradiologic correlations in a patient with familial fatal insomnia. DESIGN: Radioclinical prospective study. We describe a patient with fatal familial insomnia and normal MRI images. Because the MRI study was performed only 4 days before the patient's death, we were able to compare radiological data with the lesions observed at the neuropathologic level. PATIENT: A 55-year-old man with familial fatal insomnia. MAIN OUTCOME MEASURE: Magnetic resonance spectroscopy combined with the measurement of apparent diffusion coefficient of water in different brain areas. RESULTS: The neuroradiological study showed, in the thalamus but not in the other brain regions studied, an increase of apparent diffusion coefficient of water and a metabolic pattern indicating gliosis. These alterations closely correlated with neuropathologic data showing an almost pure gliosis that was restricted to the thalami. CONCLUSION: Considering fatal familial insomnia as a model of thalamic restricted gliosis, this case demonstrates that multisequences of magnetic resonance can detect prion-induced gliosis in vivo, as confirmed by a neuropathologic examination performed only a few days after radiological examination. PMID- 18413482 TI - Disruption of sodium bicarbonate transporter SLC4A10 in a patient with complex partial epilepsy and mental retardation. AB - OBJECTIVE: To determine gene(s) disrupted in a patient with partial frontal lobe epilepsy and cognitive impairment with concomitant de novo balanced chromosomal translocation t(2;13)(q24;q31). DESIGN: Fluorescence in situ hybridization and array comparative genomic hybridization were used to map the locations of chromosomal translocation breakpoints. RESULTS: SLC4A10 (OMIM 605556), a sodium bicarbonate transporter gene with high expression in the cerebral cortex and hippocampus, was disrupted by the translocation breakpoint on chromosome 2q24. The breakpoint on chromosome 13q31 was in a 1-megabase (Mb)-gene desert. Genomewide array comparative genomic hybridization confirmed the absence of additional chromosomal abnormalities. CONCLUSION: SLC4A10 is the third SLC4 base transporter family member to be implicated in human cognition and epilepsy. PMID- 18413483 TI - Gender and blood pressure control. PMID- 18413484 TI - Intrarenal perfusion and angiotensin II levels regulate in vivo angiotensin II type 1 receptor imaging in the kidney. PMID- 18413485 TI - Orthostatic hypercoagulability: a novel physiological mechanism to activate the coagulation system. AB - Orthostatic stress causes significant plasma shift and raises transmural pressure in lower extremities, resulting in an increase in endothelial activation and plasma proteins concentrations, possibly including coagulation factors. This may lead to activation of the coagulation system during standing. To test this hypothesis, we recruited 18 healthy volunteers (9 females and 9 males; mean age: 25+/-1.2 years; body mass index: 21.7+/-0.5 kg/m(2)). Hemodynamics, plasma shift (extrapolated from sequential hematocrit concentration), plasma proteins, and coagulation tests, including procoagulants; fibrinogen, factor V, and factor VIII activity; prothrombin fragments 1 and 2; and endothelial activation-related factors (tissue factor and von Willebrand factor), as well as protein C global pathway, were determined at rest supine and at 15 minutes, 30 minutes, and 60 minutes of still standing. Thirty minutes of standing caused a decrease in plasma volume by 12.0+/-0.5% and an increase in plasma protein by 13.0+/-0.7%. Fibrinogen, factor V, and factor VIII activity rose by 12.0+/-1.2%, 13.0+/-1.0%, and 40.0+/-6.0% (P<0.002 for all), respectively. Prothrombin fragments 1 and 2 were elevated by 150.0+/-30.0%. Tissue factor and von Willebrand factor increased by 30.0+/-9.0% and 17.4+/-51.0% (P<0.02 for both), respectively. However, protein C assay results decreased from 0.95+/-0.20 to 0.83+/-0.16 (P<0.001). We hereby introduce a novel physiological mechanism, "orthostatic procoagulation," that should be considered during coagulation tests. Furthermore, it could be extrapolated to the pathophysiology of stasis and venous thromboembolism. PMID- 18413486 TI - Angiotensin-converting enzyme inhibitors and angioedema: estimating the risk. PMID- 18413487 TI - Angiotensin II in the elderly: impact of angiotensin II type 1 receptor sensitivity on peripheral hemodynamics. AB - Exercise hyperemia is attenuated in the elderly, which may be attributed to local vasoregulatory pathways within the skeletal muscle vasculature. Therefore, we sought to determine whether healthy aging is associated with changes in angiotensin II (Ang II) receptor sensitivity through measurements of leg blood flow in resting and exercising skeletal muscle. In 12 (n=6 young, 24+/-1 years; n=6 older, 68+/-3 years) healthy volunteers, we determined changes in leg blood flow (ultrasound Doppler) before and during intra-arterial infusion of Ang II (0.8 ng/mL of leg blood flow per minute). Heart rate, arterial blood pressure, common femoral artery diameter, and mean blood velocity were measured at rest and during knee-extensor exercise at 20% and 40% of the maximal work rate (WR(max)). At rest, Ang II infusion decreased leg blood flow to a greater extent in older ( 61+/-8%) subjects compared with younger subjects (-31+/-5%). Compared with rest, Ang II-mediated vasoconstriction (leg blood flow) during exercise was diminished in both older and younger subjects at 20% (older: -7+/-5%; younger: -21+/-2%) and 40% WR(max) (older: -5+/-4%; younger: -9+/-3%). These data identify a clear age related hypersensitivity to Ang II in the resting leg, which may contribute to the recognized decrement in leg blood flow in this cohort. However, the diminished vasoconstriction to Ang II during exercise suggests that the elevation in Ang II type 1 receptor sensitivity documented at rest does not contribute significantly to the blunted exercise hyperemia experienced with advancing age. PMID- 18413488 TI - Angioedema incidence in US veterans initiating angiotensin-converting enzyme inhibitors. AB - Angioedema is a rare but potentially serious complication of angiotensin converting enzyme inhibitor (ACE) use. We conducted a study to estimate incidence of ACE-related angioedema and explore its determinants in a large racially diverse patient population. We used linked medical and pharmacy records to identify all patients in the US Veterans Affairs Health Care System from April 1999 through December 2000 who received first prescriptions for antihypertensive medications. We studied 195 192 ACE initiators and 399 889 patients initiating other antihypertensive medications (OAH). New angioedema was identified by diagnosis codes using methods validated in a national sample of 869 angioedema cases with confirmation for over 95% of cases. Overall, 0.20% of ACE initiators developed angioedema while on the medication and the incidence rate was 1.97 (1.77 to 2.18) cases per 1000 person years. This compares with a rate of 0.51 (0.43 to 0.59) in OAH initiators and the adjusted relative risk estimate was 3.56 (2.82 to 4.44). Fifty five percent of cases occurred within 90 days of first ACE use but risk remained elevated with prolonged use, even beyond 1 year. We estimate that 58.3% of angioedema in patients starting antihypertensives was related to ACE. We also found that angioedema rates were nearly 4-fold higher in blacks, 50% higher in women, and 12% lower in those with diabetes. This study provides a reliable estimate of angioedema incidence associated with ACE use in a diverse nontrial patient population, confirming that the incidence is low, but finding substantial variation by race, sex, and diabetes status. PMID- 18413489 TI - C-reactive protein is elevated 30 years after eclamptic pregnancy. AB - Women with a history of preeclampsia or eclampsia (seizure during preeclamptic pregnancy) are at increased risk for cardiovascular disease after pregnancy for reasons that remain unclear. Prospective studies during pregnancy suggest that inflammation, dyslipidemia, and insulin resistance are associated with increased risk of preeclampsia. Elevated serum C-reactive protein (CRP >3 mg/L) is an indicator of inflammation and cardiovascular risk. We hypothesized that Icelandic postmenopausal women with a history of eclampsia would manifest higher concentrations of serum CRP than Icelandic postmenopausal controls with a history of uncomplicated pregnancies. We also asked whether elevated CRP is associated with the dyslipidemia and insulin resistance previously identified in this cohort. CRP, measured by high-sensitivity enzyme-linked immunoassay, was higher in women with prior eclampsia (n=25) than controls (n=28) (median mg/L [interquartile range]: 9.0 [0.9 to 13.2] versus 2.0 [0.3 to 5.1]; P<0.03). This difference remained significant after adjustment for body mass index, smoking, hormone replacement, and current age. Women with prior eclampsia clustered into either high CRP (range 8.97 to 40.6 mg/L, n=13) or lower CRP (median 1.0, range 0.05 to 3.77, n=12) subsets. The prior eclampsia/high CRP subset displayed significantly elevated systolic blood pressures, lower high-density lipoprotein (HDL) cholesterol, higher apolipoprotein B, and higher fasting insulin and homeostasis model of insulin resistance (HOMA) values compared to controls, whereas the prior eclampsia/low CRP subset differed from controls only by marginally increased apolipoprotein B. The triad of inflammation, low HDL, and insulin resistance may elevate risk for both preeclampsia/eclampsia and cardiovascular disease in later life. PMID- 18413491 TI - Blood pressure and renal sodium handling in relation to genetic variation in the DRD1 promoter and GRK4. AB - Activation of type-1 dopamine receptors (DRD1) reduces renal sodium reabsorption. In a family-based random sample of 611 untreated whites (women, 45.0%; mean age, 38.6 years), we measured blood pressure (BP). We used the endogenous lithium clearance to assess fractional sodium excretion (FE(Na)) and proximal (RNa(prox)) and distal (RNa(dist)) tubular sodium reabsorption. We investigated multivariate adjusted associations with the DRD1 promoter (A-48G, G-94A, and C-800T) and GRK4 (Ala142Val). The frequent DRD1 haplotypes were AGC (48.2%), GGT (34.4%), and AAC (14.3%). While standardizing to mean sodium excretion (8.7 mmol/h) and adjusting for covariates and relatedness, RNa(dist) was lower in DRD1 -94GG homozygotes than -94A allele carriers (effect size, -0.94%; P=0.005) with opposite findings for FE(Na) (+0.084%; P=0.014). AGC carriers (-0.88%; P=0.012) and AAC carriers (+1.00%; P=0.004) had different RNa(dist) compared to corresponding noncarriers. Furthermore, FE(Na) was lower in AAC carriers than in noncarriers (-0.082%; P=0.019). The family-based analyses identified a significant between-family component in the variance of the renal phenotypes associated with the DRD1 polymorphisms. Transmission of the DRD1 AGC haplotype was also associated with lower systolic (-3.54 mm Hg; P=0.016) and diastolic (-2.80 mm Hg; P=0.0064) BPs without significant between-family variance component. Plasma renin activity and urinary aldosterone excretion were not associated with DRD1 variation. The GRK4 Ala142Val polymorphism did not contribute to the phenotypes under study. In conclusion, renal sodium handling and BP were associated with genetic variation in the DRD1 promoter. The between-family variance component excluded population stratification for BP, but not for the renal phenotypes. PMID- 18413490 TI - Pharmacological inhibition of epsilon-protein kinase C attenuates cardiac fibrosis and dysfunction in hypertension-induced heart failure. AB - Studies on genetically manipulated mice suggest a role for epsilon-protein kinase C (epsilonPKC) in cardiac hypertrophy and in heart failure. The potential clinical relevance of these findings was tested here using a pharmacological inhibitor of epsilonPKC activity during the progression to heart failure in hypertensive Dahl rats. Dahl rats, fed an 8% high-salt diet from the age of 6 weeks, exhibited compensatory cardiac hypertrophy by 11 weeks, followed by heart failure at approximately 17 weeks and death by the age of approximately 20 weeks (123+/-3 days). Sustained treatment between weeks 11 and 17 with the selective epsilonPKC inhibitor epsilonV1-2 or with an angiotensin II receptor blocker olmesartan prolonged animal survival by approximately 5 weeks (epsilonV1-2: 154+/ 7 days; olmesartan: 149+/-5 days). These treatments resulted in improved fractional shortening (epsilonV1-2: 58+/-2%; olmesartan: 53+/-2%; saline: 41+/ 6%) and decreased cardiac parenchymal fibrosis when measured at 17 weeks without lowering blood pressure at any time during the treatment. Combined treatment with epsilonV1-2, together with olmesartan, prolonged animal survival by 5 weeks (37 days) relative to olmesartan alone (from 160+/-5 to 197+/-14 days, respectively) and by approximately 11 weeks (74 days) on average relative to saline-treated animals, suggesting that the pathway inhibited by epsilonPKC inhibition is not identical to the olmesartan-induced effect. These data suggest that an epsilonPKC selective inhibitor such as epsilonV1-2 may have a potential in augmenting current therapeutic strategies for the treatment of heart failure in humans. PMID- 18413492 TI - Nuclear hormone receptors as regulators of the renin-angiotensin-aldosterone system. PMID- 18413493 TI - The collecting duct is the major source of prorenin in diabetes. AB - In addition to the juxtaglomerular apparatus, renin is also synthesized in renal tubular epithelium, including the collecting duct (CD). Angiotensin (Ang) II differentially regulates the synthesis of juxtaglomerular (inhibition) and CD (stimulation) renin. Because diabetes mellitus, a disease with high intrarenal renin-Ang system and Ang II activity, is characterized by high prorenin levels, we hypothesized that the CD is the major source of prorenin in diabetes. Renin granular content was visualized using in vivo multiphoton microscopy of the kidney in diabetic Munich-Wistar rats. Diabetes caused a 3.5-fold increase in CD renin, in contrast to less pronounced juxtaglomerular changes. Ang II type 1 receptor blockade with Olmesartan reduced CD renin to control levels but significantly increased juxtaglomerular renin. Using a fluorogenic renin assay, the prorenin component of CD renin content was measured by assessing the difference in enzymatic activity of medullary homogenates before and after trypsin activation of prorenin. Trypsinization caused no change in control renin activity but a 5-fold increase in diabetes. Studies on a CD cell line (M1) showed a 22-fold increase in renin activity after trypsinization and a further 35-fold increase with Ang II treatment. Therefore, prorenin significantly contributes to baseline CD renin. Diabetes, possibly via Ang II, greatly stimulates CD prorenin and causes hyperplasia of renin-producing connecting segments. These novel findings suggest that, in a rat model of diabetes, prorenin content and release from the CD may be more important than the juxtaglomerular apparatus in contrast to the existing paradigm. PMID- 18413494 TI - Hereditary determinants of human hypertension: strategies in the setting of genetic complexity. PMID- 18413495 TI - Long-term mortality after percutaneous coronary intervention with drug-eluting stent implantation versus coronary artery bypass surgery for the treatment of multivessel coronary artery disease. AB - BACKGROUND: Although previous studies have compared the treatment effects of percutaneous coronary intervention and coronary artery bypass grafting (CABG), the long-term outcomes beyond 1 year among patients with multivessel coronary artery disease who underwent percutaneous coronary intervention with drug-eluting stents (DES) or CABG have not been evaluated. METHODS AND RESULTS: Between January 2003 and December 2005, 3042 patients with multivessel disease underwent coronary implantation of DES (n=1547) or CABG (n=1495). The primary end point was all-cause mortality. In a crude analysis, the rate of long-term mortality was significantly higher in patients who underwent CABG than in those who underwent DES implantation (3-year unadjusted mortality rate, 7.0% for CABG versus 4.4% for percutaneous coronary intervention; P=0.01). However, after adjustment for baseline differences, the overall risks of death were similar among all patients (hazard ratio, 0.85; 95% confidence interval [CI], 0.56 to 1.30; P=0.45), diabetic patients (hazard ratio, 1.76; 95% CI, 0.82 to 3.78; P=0.15), and patients with compromised ventricular function (hazard ratio, 1.39; 95% CI, 0.41 to 4.65; P=0.60). In the anatomic subgroups, mortality benefit with DES implantation was noted in patients with 2-vessel disease with involvement of the nonproximal left anterior descending artery (hazard ratio, 0.23; 95% CI, 0.01 to 0.78; P=0.016). The rate of revascularization was significantly higher in the DES than in the CABG group (hazard ratio, 2.81; 95% CI, 2.11 to 3.75; P<0.001). CONCLUSIONS: For the treatment of multivessel coronary artery disease, percutaneous coronary intervention with DES implantation showed equivalent long term mortality as CABG. PMID- 18413496 TI - Increased high-density lipoprotein cholesterol predicts the pioglitazone-mediated reduction of carotid intima-media thickness progression in patients with type 2 diabetes mellitus. AB - BACKGROUND: Measurement of carotid intima-media thickness (CIMT) has been validated as a measure of atherosclerosis and as a predictor of future cardiovascular events. Compared with glimepiride, pioglitazone has been shown to slow the progression of atherosclerosis measured by CIMT in patients with type 2 diabetes mellitus. METHODS AND RESULTS: We evaluated individual cardiovascular risk factors as predictors of the change in CIMT produced by pioglitazone treatment by determining whether their addition to a baseline model resulted in loss of significance for the treatment effect on CIMT. Pioglitazone treatment led to improvement in levels of multiple cardiovascular risk markers, including high sensitivity C-reactive protein, apolipoprotein B, apolipoprotein A1, high-density lipoprotein (HDL) cholesterol, triglyceride, insulin, and free fatty acid. At 24 weeks, there were significant differences in HDL cholesterol, triglyceride, total cholesterol, low-density lipoprotein cholesterol, insulin, body mass index, hip circumference, and high-sensitivity C-reactive protein between the pioglitazone and glimepiride treatment groups. After adjustment for 24-week on-treatment values of cardiovascular risk factors, only inclusion of the changes in HDL cholesterol and insulin significantly impacted the magnitude and significance of the treatment effect on CIMT. Furthermore, irrespective of treatment assignment, increased HDL cholesterol at 24 weeks was a significant predictor of reduced CIMT progression at 72 weeks. CONCLUSIONS: The beneficial effect of pioglitazone on HDL cholesterol at 24 weeks predicted its beneficial effect for reducing CIMT progression at 72 weeks. Changes in HDL cholesterol at 24 weeks, irrespective of treatment, predicted less progression of CIMT at 72 weeks. These results suggest that suppression of atherosclerosis with pioglitazone therapy is linked to its ability to raise HDL cholesterol. PMID- 18413497 TI - Local delivery of gene vectors from bare-metal stents by use of a biodegradable synthetic complex inhibits in-stent restenosis in rat carotid arteries. AB - BACKGROUND: Local drug delivery from polymer-coated stents has demonstrated efficacy for preventing in-stent restenosis; however, both the inflammatory effects of polymer coatings and concerns about late outcomes of drug-eluting stent use indicate the need to investigate innovative approaches, such as combining localized gene therapy with stent angioplasty. Thus, we investigated the hypothesis that adenoviral vectors (Ad) could be delivered from the bare metal surfaces of stents with a synthetic complex for reversible vector binding. METHODS AND RESULTS: We synthesized the 3 components of a gene vector binding complex: (1) A polyallylamine bisphosphonate with latent thiol groups (PABT), (2) a polyethyleneimine (PEI) with pyridyldithio groups for amplification of attachment sites [PEI(PDT)], and (3) a bifunctional (amine- and thiol-reactive) cross-linker with a labile ester bond (HL). HL-modified Ad attached to PABT/PEI(PDT)-treated steel surfaces demonstrated both sustained release in vitro over 30 days and localized green fluorescent protein expression in rat arterial smooth muscle cell cultures, which were not sensitive to either inhibition by neutralizing anti-Ad antibodies or inactivation after storage at 37 degrees C. In rat carotid studies, deployment of steel stents configured with PABT/PEI(PDT)/HL tethered adenoviral vectors demonstrated both site-specific arterial Ad(GFP) expression and adenovirus-luciferase transgene activity per optical imaging. Rat carotid stent delivery of adenovirus encoding inducible nitric oxide synthase resulted in significant inhibition of restenosis. CONCLUSIONS: Reversible immobilization of adenovirus vectors on the bare-metal surfaces of endovascular stents via a synthetic complex represents an efficient, tunable method for sustained release of gene vectors to the vasculature. PMID- 18413498 TI - Spontaneous myocardial infarction in mice lacking all nitric oxide synthase isoforms. AB - BACKGROUND: The roles of nitric oxide (NO) in the cardiovascular system have been investigated extensively in pharmacological studies with NO synthase (NOS) inhibitors and in studies with NOS isoform-deficient mice. However, because of the nonspecificity of the NOS inhibitors and the compensatory interactions among NOS isoforms (nNOS, iNOS, and eNOS), the ultimate roles of endogenous NO derived from the entire NOS system are still poorly understood. In this study, we examined this point in mice deficient in all 3 NOS isoforms (triply n/i/eNOS(-/-) mice) that we have recently developed. METHODS AND RESULTS: The triply n/i/eNOS( /-) mice, but not singly eNOS(-/-) mice, exhibited markedly reduced survival, possibly due to spontaneous myocardial infarction accompanied by severe coronary arteriosclerotic lesions. Furthermore, the triply n/i/eNOS(-/-) mice manifested phenotypes that resembled metabolic syndrome in humans, including visceral obesity, hypertension, hypertriglyceridemia, and impaired glucose tolerance. Importantly, activation of the renin-angiotensin system was noted in the triply n/i/eNOS(-/-) mice, and long-term oral treatment with an angiotensin II type 1 receptor blocker significantly suppressed coronary arteriosclerotic lesion formation and the occurrence of spontaneous myocardial infarction and improved the prognosis of those mice, along with ameliorating the metabolic abnormalities. CONCLUSIONS: These results provide the first direct evidence that genetic disruption of the whole NOS system causes spontaneous myocardial infarction associated with multiple cardiovascular risk factors of metabolic origin in mice in vivo through the angiotensin II type 1 receptor pathway, demonstrating the critical role of the endogenous NOS system in maintaining cardiovascular and metabolic homeostasis. PMID- 18413499 TI - Inflammation, oxidative stress, and repair capacity of the vascular endothelium in obstructive sleep apnea. AB - BACKGROUND: Indirect evidence implicates endothelial dysfunction in the pathogenesis of vascular diseases associated with obstructive sleep apnea (OSA). We investigated directly whether dysfunction and inflammation occur in vivo in the vascular endothelium of patients with OSA. The effects of continuous positive airway pressure (CPAP) therapy on endothelial function and repair capacity were assessed. METHODS AND RESULTS: Thirty-two patients with newly diagnosed OSA and 15 control subjects were studied. Proteins that regulate basal endothelial nitric oxide (NO) production (endothelial NO synthase [eNOS] and phosphorylated eNOS) and inflammation (cyclooxygenase-2 and inducible NOS) and markers of oxidative stress (nitrotyrosine) were quantified by immunofluorescence in freshly harvested venous endothelial cells before and after 4 weeks of CPAP therapy. Vascular reactivity was measured by flow-mediated dilation. Circulating endothelial progenitor cell levels were quantified to assess endothelial repair capacity. Baseline endothelial expression of eNOS and phosphorylated eNOS was reduced by 59% and 94%, respectively, in patients with OSA compared with control subjects. Expression of both nitrotyrosine and cyclooxygenase-2 was 5-fold greater in patients with OSA than in control subjects, whereas inducible NOS expression was 56% greater. Expression of eNOS and phosphorylated eNOS significantly increased, whereas expression of nitrotyrosine, cyclooxygenase-2, and inducible NOS significantly decreased in patients who adhered to CPAP > or = 4 hours daily. Baseline flow-mediated dilation and endothelial progenitor cell levels were lower in patients than in control subjects, and both significantly increased in patients who adhered to CPAP > or = 4 hours daily. CONCLUSIONS: OSA directly affects the vascular endothelium by promoting inflammation and oxidative stress while decreasing NO availability and repair capacity. Effective CPAP therapy is associated with the reversal of these alterations. PMID- 18413500 TI - Genetic components of the circadian clock regulate thrombogenesis in vivo. AB - BACKGROUND: Myocardial infarction, stroke, and sudden death undergo diurnal variation. Although genes relevant to hemostasis and vascular integrity undergo circadian oscillation, the role of the molecular clock in thrombotic events remains to be established. METHODS AND RESULTS: A diurnal variation in the time to thrombotic vascular occlusion (TTVO) subsequent to a photochemical injury was observed in wild-type mice: TTVO varied from 24.6+/-2.7 minutes at zeitgeber time (ZT) 2 to 40.3+/-4.3 minutes at ZT8, 24.3+/-2.3 minutes at ZT14, and 31.0+/-4.4 minutes at ZT20. This pattern was disrupted or altered when core clock genes BMAL1, CLOCK, and NPAS2-were mutated or deleted. Mutation of CLOCK abolished the diurnal variation in TTVO, whereas deletion of NPAS2 altered its temporal pattern. NPAS2 deletion prolonged TTVO and reduced blood pressure irrespective of clock time. Global BMAL1 deletion shortened TTVO at ZT8, and the diurnal variation in TTVO, but not in systemic blood pressure, was disrupted in mice in which BMAL1 had been selectively deleted in endothelium. CONCLUSIONS: Key components of the molecular clock regulate the response to a thrombogenic stimulus in vivo. Such a phenomenon may interact with environmental variables, and together with the influence of these genes on blood pressure may contribute to the diurnal variation in cardiovascular events observed in humans. PMID- 18413501 TI - Effects of off-pump versus on-pump coronary artery bypass grafting on early and late right ventricular function. AB - BACKGROUND: Off-pump CABG (OPCABG) results in better preservation of left ventricular function in the perioperative period than conventional on-pump CABG (ONCABG); however, evidence is conflicting as to the effect of OPCABG and ONCABG on right ventricular (RV) function, possibly because of the complexity involved in measuring this. METHODS AND RESULTS: In a single-center randomized pilot study, 60 patients with normal left ventricular function undergoing CABG were randomly assigned to OPCABG or ONCABG. Patients underwent cardiac magnetic resonance imagine for assessment of RV function preoperatively, early postoperatively, and at 6 months after surgery. Fifty-one patients completed the first 2 scans, and 47 completed all 3 scans. Preoperative characteristics and RV function did not differ significantly between the 2 groups (mean+/-SD): RV stroke volume index was 49+/-10 mL/m(2) for OPCABG and 49+/-16 mL/m(2) for ONCABG. After surgery, RV stroke volume index fell to 36+/-7 mL/m(2) in the OPCABG group and 39+/-11 mL/m(2) in the ONCABG group, but this did not differ significantly between the 2 groups (P=0.41). All markers of RV function recovered to preoperative levels by 6 months, with no long-term difference between the surgical techniques. CONCLUSIONS: RV function is impaired early after surgery but recovers by 6 months. The changes were similar in both the OPCABG and ONCABG groups. PMID- 18413502 TI - Role of left ventricular stiffness in heart failure with normal ejection fraction. AB - BACKGROUND: Increased left ventricular stiffness is a distinct finding in patients who have heart failure with normal ejection fraction (HFNEF). To elucidate how diastolic dysfunction contributes to heart failure symptomatology during exercise, we conducted a study using an invasive pressure-volume loop approach and measured cardiac function at rest and during atrial pacing and handgrip exercise. METHODS AND RESULTS: Patients with HFNEF (n=70) and patients without heart failure symptoms (n=20) were enrolled. Pressure-volume loops were measured with a conductance catheter during basal conditions, handgrip exercise, and atrial pacing with 120 bpm to analyze diastolic and systolic left ventricular function. During transient preload reduction, the diastolic stiffness constant was measured directly. Diastolic function with increased stiffness was significantly impaired in patients with HFNEF during basal conditions. This was associated with increased end-diastolic pressures during handgrip exercise and with decreased stroke volume and a leftward shift of pressure-volume loops during atrial pacing. CONCLUSIONS: Increased left ventricular stiffness contributed to increased end-diastolic pressure during handgrip exercise and decreased stroke volume during atrial pacing in patients with HFNEF. These data suggest that left ventricular stiffness modulates cardiac function in HFNEF patients and suggests that diastolic dysfunction with increased stiffness is a target for treating HFNEF. PMID- 18413503 TI - Essential features of designating out-of-hospital cardiac arrest as a reportable event: a scientific statement from the American Heart Association Emergency Cardiovascular Care Committee; Council on Cardiopulmonary, Perioperative, and Critical Care; Council on Cardiovascular Nursing; Council on Clinical Cardiology; and Quality of Care and Outcomes Research Interdisciplinary Working Group. AB - The 2010 impact goal of the American Heart Association is to reduce death rates from heart disease and stroke by 25% and to lower the prevalence of the leading risk factors by the same proportion. Much of the burden of acute heart disease is initially experienced out of hospital and can be reduced by timely delivery of effective prehospital emergency care. Many patients with an acute myocardial infarction die from cardiac arrest before they reach the hospital. A small proportion of those with cardiac arrest who reach the hospital survive to discharge. Current health surveillance systems cannot determine the burden of acute cardiovascular illness in the prehospital setting nor make progress toward reducing that burden without improved surveillance mechanisms. Accordingly, the goals of this article provide a brief overview of strategies for managing out-of hospital cardiac arrest. We review existing surveillance systems for monitoring progress in reducing the burden of out-of-hospital cardiac arrest in the United States and make recommendations for filling significant gaps in these systems, including the following: 1. Out-of-hospital cardiac arrests and their outcomes through hospital discharge should be classified as reportable events as part of a heart disease and stroke surveillance system. 2. Data collected on patients' encounters with emergency medical services systems should include descriptions of the performance of cardiopulmonary resuscitation by bystanders and defibrillation by lay responders. 3. National annual reports on key indicators of progress in managing acute cardiovascular events in the out-of-hospital setting should be developed and made publicly available. Potential barriers to action on cardiac arrest include concerns about privacy, methodological challenges, and costs associated with designating cardiac arrest as a reportable event. PMID- 18413504 TI - Images in cardiovascular medicine. Sinus of Valsalva aneurysms: a unique case of giant aneurysms involving all 3 sinuses. PMID- 18413505 TI - Letter by Violi and Pignatelli regarding article, "Effects of random allocation to vitamin E supplementation on the occurrence of venous thromboembolism: report from the Women's Health Study". PMID- 18413507 TI - Life versus death. PMID- 18413508 TI - Life and death in Denmark: lessons about diabetes and coronary heart disease. PMID- 18413509 TI - Patient selection and echocardiographic assessment of dyssynchrony in cardiac resynchronization therapy. PMID- 18413510 TI - Frontiers in intravascular imaging technologies. PMID- 18413511 TI - Images in cardiovascular medicine. Sudden cardiac death due to triple vessel coronary dissection. PMID- 18413512 TI - Histologic analysis of descemet stripping in posterior lamellar keratoplasty. AB - OBJECTIVE: To investigate how precise Descemet stripping works in posterior lamellar keratoplasty (Descemet stripping automated endothelial keratoplasty [DSAEK]) for the treatment of corneal endothelial disorders. METHODS: In a prospective, single-center, nonrandomized consecutive series, 20 Descemet membrane specimens obtained after Descemet stripping in DSAEK using a Price hook were examined using histologic analysis and transmission electron microscopy for the presence of residual stroma, thickness of the Descemet membrane, endothelial cell count, and presence of guttae or a posterior collagenous layer. Pathologic findings were correlated with the underlying clinical disease. RESULTS: Light and electron microscopy revealed no evidence of adherent rests of corneal stroma in all 20 specimens after Descemet stripping. The mean (SD) total thickness of the Descemet membrane was 21.5 (4.5) microm in peripheral localization and 17.6 (3.8) microm in central localization. The anterior banded layer measured a mean (SD) of 3.0 (0.8) microm thick; the posterior nonbanded layer, 16.7 (5.2) microm thick. The mean (SD) endothelial cell count was 1.7 (1.4) cells per high-power field. Guttae were seen in 15 specimens (75%), and a posterior collagenous layer was found in 3 (15%). CONCLUSION: Descemet stripping in DSAEK using the Price hook achieves complete and specific removal of the Descemet membrane without adherent stroma in different underlying endothelial pathologic abnormalities. PMID- 18413514 TI - Logistic regression analysis for early glaucoma diagnosis using optical coherence tomography. AB - OBJECTIVE: To determine and validate the diagnostic ability of a linear discriminant function (LDF) based on the retinal nerve fiber layer thickness at each of the 12 clock-hour positions obtained using optical coherence tomography for discriminating between healthy eyes and eyes with early glaucomatous visual field loss. METHODS: We prospectively selected 62 consecutive healthy individuals and 73 patients with open-angle glaucoma to calculate the LDF. Another independent prospective sample of 280 healthy eyes and 302 glaucomatous eyes was used to evaluate the diagnostic accuracy of the LDF. RESULTS: The proposed function was LDF = 15.584 - (12-o'clock segment thickness x 0.032) - (7-o'clock segment thickness x 0.041) - (3-o'clock segment thickness [nasal side] x 0.121). The greatest area under the receiver operating characteristic curve was observed for our LDF in both populations: 0.962 and 0.922. Our LDF and the average thickness yielded sensitivities of 74.5% and 67.8%, respectively, at a fixed specificity of 95%. CONCLUSIONS: The LDF increased the diagnostic ability of the isolated retinal nerve fiber layer thickness at the 12 clock-hour positions. Compared with optical coherence tomography-provided parameters, our LDF had the highest sensitivities at 85% and 95% fixed specificities to discriminate between healthy and early glaucomatous eyes. PMID- 18413515 TI - Total deviation probability plots for stimulus size v perimetry: a comparison with size III stimuli. AB - OBJECTIVE: To compare empirical probability plots in patients with glaucoma for size V and III perimetry testing. METHODS: We computed empirical probability plot percentile limits after testing 60 age-matched controls tested with both size III (Swedish interactive thresholding algorithm) and size V (full threshold) perimetry twice. Probability plots of 120 patients with glaucoma tested in the same way were computed. We compared the number of abnormal test locations in the 2 stimulus sizes; we then compared these results with those from size III StatPac software (Zeiss Humphrey Systems, Dublin, California) using 2-way repeated measures analysis of variance. RESULTS: We found a similar number of abnormal test locations (P < or = .05) for the size III and size V testing conditions identified by the probability plots (no significant difference); there were significantly fewer abnormal locations using StatPac (size III) than from our size III database. When results were stratified by mean deviation, the mild visual loss group again did not show any significant differences between sizes III and V. CONCLUSIONS: Size V full-threshold testing gives a similar number of abnormal test locations in patients with glaucoma compared with the size III Swedish interactive thresholding algorithm standard test. Size V testing, with its greater dynamic range and lower variability, may be a viable alternative to size III testing in patients with glaucoma. PMID- 18413516 TI - Surgical treatment of strabismus secondary to glaucoma drainage device. AB - OBJECTIVE: To describe surgical strategies in a series of patients with diplopia following implantation of a glaucoma drainage device. METHODS: Retrospective review of 9 consecutive patients who underwent strabismus surgery because of strabismus and diplopia after implantation of a glaucoma drainage device. RESULTS: Seven patients with marked limitation to ocular rotations and incomitant strabismus underwent surgery on the eye with the implant. Two patients with mild limitation to ocular rotations of the involved eye underwent surgery on the contralateral eye. All patients had a large fibrous capsule surrounding the implant plate, adjacent muscles, and sclera. Intraocular pressure was not elevated postoperatively. Postoperative diplopia in the primary position was eliminated in 5 patients and markedly improved in 3 patients. CONCLUSIONS: Strabismus following implantation of a glaucoma drainage device is an uncommon but serious complication. Restoration of ocular alignment is a complex undertaking requiring strabismus and glaucoma surgical expertise. Multiple surgical complications may occur. Surgical intervention may require complete removal of the fibrous capsule surrounding the implant and involved adjacent structures. Size reduction of the implant plate is helpful and did not interfere with postoperative intraocular pressure control in this study. Surgery on the contralateral eye is an option in patients with mild restriction. PMID- 18413517 TI - Preverbal photoscreening for amblyogenic factors and outcomes in amblyopia treatment: early objective screening and visual acuities. AB - BACKGROUND: Previous studies have suggested that infant photoscreening yields better results than visual acuity screening in preschool-aged children. With conventional vision screening, the patient must be able to provide monocular visual acuity cooperation, whereas objective screening for amblyogenic factors can be done at much younger ages. METHODS: From February 1996 through February 2006, Alaska Blind Child Discovery photoscreened 21,367 rural and urban Alaskan children through grade 2, with an 82% positive predictive value (ie, true number of those referred); 6.9% were referred for a complete eye examination and treatment. All "referred" interpreted images for children younger than 48 months who were then followed up and treated for more than 2 years were reviewed to determine whether treatment was successful. RESULTS: Of 411 "positive" screening photos from children younger than 4 years, 94 patients had more than 2 years follow-up. The 36 children photoscreened before age 2 years had a mean treated visual acuity of 0.17 logarithm of the minimum angle of resolution (logMAR), which was significantly better than that of 58 children screened between ages 25 and 48 months (mean, 0.26 logMAR). Despite similar levels of amblyogenic risk factors, the proportion of children failing to reach a visual acuity of 20/40 was significantly less among those screened before age 2 years (5%) than in those screened from ages older than 2.0 years and younger than 4.0 years (17%). CONCLUSION: Very early photoscreening yields better visual outcomes in amblyopia treatment compared with later photoscreening in preschool-aged children. PMID- 18413518 TI - Cost-utility analysis of telemedicine and ophthalmoscopy for retinopathy of prematurity management. AB - OBJECTIVE: To evaluate the cost-effectiveness of telemedicine and standard ophthalmoscopy for retinopathy of prematurity (ROP) management. METHODS: Models were developed to represent ROP examination and treatment using telemedicine and standard ophthalmoscopy. Cost-utility analysis was performed using decision analysis, evidence-based outcome data from published literature, and present value modeling. Visual outcome data were converted to patient preference-based time trade-off utility values based on published literature. Costs of disease management were determined based on 2006 Medicare reimbursements. Costs per quality-adjusted life year gained by telemedicine and ophthalmoscopy for ROP management were compared. One-way sensitivity analysis was performed on the following variables: discount rate (0%-7%), incidence of treatment-requiring ROP (1%-20%), sensitivity and specificity of ophthalmoscopic diagnosis (75%-100%), percentage of readable telemedicine images (75%-100%), and sensitivity and specificity of telemedicine diagnosis (75%-100%). RESULTS: For infants with birth weight less than 1500 g using a 3% discount rate for costs and outcomes, the costs per quality-adjusted life year gained were $3193 with telemedicine and $5617 with standard ophthalmoscopy. Sensitivity analysis resulted in ranges of costs per quality-adjusted life year from $1235 to $18,898 for telemedicine and from $2171 to $27,215 for ophthalmoscopy. CONCLUSIONS: Telemedicine is more cost effective than standard ophthalmoscopy for ROP management. Both strategies are highly cost-effective compared with other health care interventions. PMID- 18413519 TI - Prevention of exuberant granulation tissue and neovascularization in the rat cornea by naltrexone. AB - OBJECTIVE: To determine whether topical application of naltrexone prevents exuberant granulation tissue formation with neovascularization in diabetic rat corneas. METHODS: Diabetes was induced with streptozotocin. A 5-mm corneal abrasion at 9 or 11 weeks was treated topically for 7 days (4 times daily) with naltrexone or a sterile vehicle. RESULTS: Within 2 to 5 days after reepithelialization, diabetic rats given the sterile vehicle had a 41% incidence of corneal lesions represented by exuberant granulation tissue with corneal neovascularization extending from the limbus. These lesions exhibited edema, cellular and vascular inflammation, and disruption of stromal lamella by fibrovascular tissue and calcium mineralization, but infection was not detected. No corneal lesions were recorded in the diabetic group treated with naltrexone or the control group given the sterile vehicle. Diabetic rats with corneal lesions given the sterile vehicle reepithelialized more slowly than diabetic rats given the sterile vehicle without such lesions, but no difference in blood glucose levels were noted. CONCLUSIONS: Using a minimally invasive model in diabetic rats, topical naltrexone normalizes corneal wound healing and prevents neovascularization. CLINICAL RELEVANCE: Direct application of naltrexone may serve as an important strategy for facilitating corneal healing and inhibiting corneal neovascularization. PMID- 18413520 TI - The foveal avascular region of developing human retina. AB - OBJECTIVE: To study the development of the perifoveal retinal vasculature. METHODS: We studied 7 retinas aged between 26 weeks' gestation and 1 week postnatal (41 weeks' gestation). Sections were imaged using high-resolution digital photography and blood vessel profiles identified at 200% to 300% magnification. Flat mounts were immunolabeled using antibodies to CD31 and factor VIII to identify blood vessels and antibodies to rhodopsin to identify the rod free zone. RESULTS: The foveal region was identified by the absence of rod photoreceptors in the outer retina and/or presence of a shallow depression in the inner retina. The whole mount at 26 weeks' gestation showed a blood vessel-free region centered on the rod-free zone that was open along the horizontal meridian on the temporal side. At 37 weeks' gestation, the foveal avascular zone formed a complete circle. In sections, the foveal avascular zone was approximately 500 microm in diameter at 35 weeks' gestation and 300 to 350 microm at 40 weeks' gestation; in whole mounts, it was 150 to 170 microm in diameter at 37 and 41 weeks' gestation. CONCLUSIONS: The foveal region is normally avascular during development, as in adult life. We found no evidence of foveal vascularization during development of the human retina. Clinical Relevance Instances of vascularization of the foveal region are not due to failed regression of a transient vasculature. PMID- 18413521 TI - The 15-year cumulative incidence of retinal vein occlusion: the Beaver Dam Eye Study. AB - OBJECTIVES: To describe the 15-year incidence of retinal vein occlusion (central retinal vein occlusion and branch retinal vein occlusion) and associated risk factors. METHODS: A population-based study where branch retinal vein occlusion and central retinal vein occlusion were detected at baseline (n = 4068, 1988 1990) and three 5-year follow-up examinations by grading 30 degrees color fundus photographs. RESULTS: The 15-year cumulative incidences of branch retinal vein occlusion and central retinal vein occlusion were 1.8% and 0.5%, respectively. Using a generalized estimating equation model, incident retinal vein occlusion was related to baseline age (odds ratio [OR] per 10 years, 1.70; 95% confidence interval [CI], 1.36-2.12), history of barbiturate use (OR, 5.30; 95% CI, 2.28 12.31), focal retinal arteriolar narrowing (OR, 2.45; 95% CI, 1.29-4.66), glaucoma (OR, 3.17; 95% CI, 1.50-6.69), serum ionized calcium level (OR per 0.4 mg/dL, 0.43; 95% CI, 0.23-0.79), serum phosphorus level (OR per 0.3 mg/dL, 1.15; 95% CI, 1.01-1.30), and serum creatinine level (OR for > or = 1.4 vs < 1.4 mg/dL, 1.61; 95% CI, 1.00-2.59). Migraine headache history was associated with branch retinal vein occlusion (OR, 1.99; 95% CI, 1.08-3.67). Diabetes history was associated with central retinal vein occlusion (OR, 6.35; 95% CI, 1.90-21.27). CONCLUSIONS: Incident retinal vein occlusion is not infrequent in the population, especially after age 65 years. The relationships of barbiturate use, serum creatinine level, serum ionized calcium level, and serum phosphorus level with incident retinal vein occlusion require further assessment in other large population-based studies. PMID- 18413522 TI - Menopausal and reproductive factors and risk of age-related macular degeneration. AB - OBJECTIVE: To investigate whether estrogen exposures are associated with lower risks of age-related macular degeneration (AMD). METHODS: Postmenopausal hormone (PMH) use, past use of oral contraceptives (OCs), ages at menarche and menopause, and parity were assessed among 74,996 postmenopausal women. Over 22 years, cases of early (n = 554) and neovascular (n = 334) AMD with a visual acuity of 20/30 or worse were identified. Cox models were used to calculate the relative risk for each exposure, adjusted for smoking and other factors. RESULTS: Current PMH users had a notable 48% lower risk of neovascular AMD compared with those who had never used PMH, although risk did not decline linearly with longer durations of use. Risk was lowest for PMH users who had used OCs in the past (P value for interaction, .03). In contrast, risk of early AMD was a notable 34% higher among current PMH users and OC use was unassociated with risk. The only remarkable finding for the endogenous estrogenic factors was a 26% lower risk of early AMD for parous women. CONCLUSIONS: Although PMH and OC use were associated with a lower risk of neovascular AMD, no benefit was observed for early AMD. Factors influencing lifetime exposure to estrogens were not consistently associated with the disease. PMID- 18413523 TI - Myopia, lifestyle, and schooling in students of Chinese ethnicity in Singapore and Sydney. AB - OBJECTIVE: To compare the prevalence and risk factors for myopia in 6- and 7-year old children of Chinese ethnicity in Sydney and Singapore. METHODS: Two cross sectional samples of age- and ethnicity-matched primary school children participated: 124 from the Sydney Myopia Study and 628 from the Singapore Cohort Study on the Risk Factors for Myopia. Cycloplegic autorefraction was used to determine myopia prevalence (spherical equivalent < or = -0.5 diopter). Lifestyle activities were ascertained by questionnaire. RESULTS: The prevalence of myopia in 6- and 7-year-old children of Chinese ethnicity was significantly lower in Sydney (3.3%) than in Singapore (29.1%) (P < .001). The prevalence of myopia in 1 or more parents was 68% in Sydney and 71% in Singapore. Children in Sydney read more books per week (P < .001) and did more total near-work activity (P = .002). Children in Sydney spent more time on outdoor activities (13.75 vs 3.05 hours per week; P < .001), which was the most significant factor associated with the differences in the prevalence of myopia between the 2 sites. CONCLUSIONS: The lower prevalence of myopia in Sydney was associated with increased hours of outdoor activities. We hypothesize that another factor contributing to the differences in the prevalence of myopia may be the early educational pressures found in Singapore but not in Sydney. PMID- 18413524 TI - Finger's amniotic membrane buffer technique: protecting the cornea during radiation plaque therapy. AB - OBJECTIVE: To use amniotic membranes as a buffer between the cornea and radioactive eye plaques. METHODS: Six melanomas were treated with ophthalmic plaque radiation therapy. Plaque-tumor localization required that a portion of the gold plaque touch the cornea during treatment. To enhance patient comfort and protect the cornea, an (0.1-mm-thick) amniotic membrane was interposed between the metal plaque edge and the cornea. RESULTS: Minimal ocular discomfort was noted during plaque radiation therapy. On a scale of 1 (none) to 10 (severe), all 6 patients reported pain levels of 1. As a tissue equivalent and because the mean thickness was only 0.1 mm, amniotic membranes had no significant effect on radiation dose calculations. No adverse effects, infections, or abrasions were noted. CONCLUSION: The amniotic membrane buffer technique improves patient comfort and protects the cornea during ophthalmic plaque radiation therapy. PMID- 18413525 TI - Imaging the ocular anterior segment with real-time, full-range Fourier-domain optical coherence tomography. AB - We have demonstrated a novel Fourier-domain optical coherence tomography system and signal-processing algorithm for full-range, real-time, artifact-free quantitative imaging of the anterior chamber. Cross-sectional full-range images comprising 1024 x 800 pixels (axial x lateral) were acquired and displayed at 6.7 images/s. Volumetric data comprising 1024 x 400 x 60 pixels (axial x lateral x elevation) were acquired in 4.5 seconds with real-time visualization of individual slices and 3-dimensional reconstruction performed in postprocessing. Details of the cornea, limbus, iris, anterior lens capsule, trabecular meshwork, and Schlemm's canal were visualized. Quantitative surface height maps of the corneal epithelium and endothelium were obtained from the volumetric data and used to generate corneal thickness maps. PMID- 18413526 TI - Small primate, big eyes. PMID- 18413527 TI - Retinal imaging with adaptive optics scanning laser ophthalmoscopy in unexplained central ring scotoma. AB - Adaptive optics scanning laser ophthalmoscopy allows for noninvasive, in vivo visualization of retinal abnormalities at a cellular level. We herein describe for the first time, to our knowledge, the utility of high-resolution retinal imaging in studying the photoreceptor mosaic in an otherwise unexplained visual disturbance. Imaging of the cone mosaic was performed in a 64-year-old man with a unilateral ringlike paracentral distortion that could not be explained using common clinical imaging instruments. Adaptive optics scanning laser ophthalmoscopy findings revealed a parafoveal circular abnormality of the cone mosaic approximately 3 degrees in diameter that corresponded to the ring of visual disturbance. Visualization of the cone mosaic with adaptive optics scanning laser ophthalmoscopy can reveal photoreceptor damage that may not be detectable with standard imaging devices. Optical axial sectioning of the retina may help in identifying and localizing abnormalities within the retinal layers. PMID- 18413528 TI - First-time failure rates of candidates for board certification: an educational outcome measure. AB - BACKGROUND: Few objective standards are available to assess the educational effectiveness of ophthalmology residency programs. As a possible measure, we evaluated the first-time failure (FTF) rate in the examinations of the American Board of Ophthalmology, defined as a first-attempt failure in the written examination or a first-attempt failure in the oral examination after having passed the written examination on the first attempt. METHOD: We tracked data on all residents who graduated between June 30, 1999, and December 31, 2003, from commencement of training to certification, including rates of overall FTF, written and oral FTF, and program FTF. Performance was analyzed for several factors, including program size. RESULTS: The FTF rate was 28% overall and ranged from 0% to 89% across 118 programs (median, 27%). Programs with fewer than 16 graduates per 5 years were significantly more likely to have higher FTF rates than larger programs. Thirty-two programs accounted for 50% of the FTF rate. CONCLUSIONS: The FTF rate is a potentially useful measure. However, the small size of many programs contributes to some imprecision. Therefore, this measure should be used in conjunction with other factors when assessing the educational effectiveness of ophthalmology residency programs. Although the eventual certification rate was high, graduates from a few programs appeared inadequately prepared to take the examinations. PMID- 18413529 TI - Treatment of postcataract extraction endophthalmitis: a summary of the results from the Endophthalmitis Vitrectomy Study. PMID- 18413530 TI - Legacy of the endophthalmitis vitrectomy study. PMID- 18413531 TI - First-time failure rates of candidates for American Board of Ophthalmology certification. PMID- 18413532 TI - Yellow dye laser treatment of vascularized corneal stromal scars in pediatric patients. PMID- 18413533 TI - NMO antibody-positive recurrent optic neuritis without clear evidence of transverse myelitis. PMID- 18413534 TI - Optical coherence tomography provides insight into the effect of intacs in keratoconus. PMID- 18413535 TI - Pulmonary metastasis masquerading as anterior uveitis. PMID- 18413536 TI - Synergistic convergence in congenital extraocular muscle misinnervation. PMID- 18413537 TI - Pituitary apoplexy causing isolated blindness after cardiac bypass surgery. PMID- 18413539 TI - Accurate calculation of longer-term incidences from short-term incidence values. PMID- 18413538 TI - Treatment of iris melanoma and secondary neovascular glaucoma using bevacizumab and plaque radiotherapy. PMID- 18413540 TI - Preenucleation radiotherapy, uveal melanoma, and competing risks. PMID- 18413545 TI - Large optic disc. PMID- 18413546 TI - Cataract surgery with primary intraocular lens implantation in children with chronic uveitis. PMID- 18413547 TI - Dark adaptation is critical for accurate pupil measurement. PMID- 18413548 TI - Length of stay and mortality in pulmonary embolism: high time for evidence-based discharge criteria. PMID- 18413549 TI - Extracolonic findings with computed tomographic colonography: asset or liability? PMID- 18413550 TI - Pharmacist care of patients with heart failure: a systematic review of randomized trials. AB - BACKGROUND: While the role of multidisciplinary teams in the treatment of patients with heart failure (HF) is well established, there is less evidence to characterize the role of individual team members. To clarify the role of pharmacists in the care of patients with HF, we performed a systematic review evaluating the effect of pharmacist care on patient outcomes in HF. METHODS: We searched PubMed, MEDLINE, EMBASE, International Pharmaceutical Abstracts, Web of Science, Scopus, Dissertation Abstracts, CINAHL, Pascal, and Cochrane Central Register of Controlled Trials for controlled studies from database inception to August 2007. We included randomized controlled trials that evaluated the impact of pharmacist care activities on patients with HF (in both inpatient and outpatient settings). Summary odds ratios (ORs) with 95% confidence intervals (CIs) were calculated using a random-effects model for rates of all-cause hospitalization, HF hospitalization, and mortality. RESULTS: A total of 12 randomized controlled trials (2060 patients) were identified. Extent of pharmacist involvement varied among studies, and each study intervention was categorized as pharmacist-directed care or pharmacist collaborative care using a priori definitions and feedback from primary study authors. Pharmacist care was associated with significant reductions in the rate of all-cause hospitalizations (11 studies [2026 patients]) (OR, 0.71; 95% CI, 0.54-0.94) and HF hospitalizations (11 studies [1977 patients]) (OR, 0.69; 95% CI, 0.51-0.94),and a nonsignificant reduction in mortality (12 studies [2060 patients])(OR, 0.84; 95% CI, 0.61-1.15). Pharmacist collaborative care led to greater reductions in the rate of HF hospitalizations (OR, 0.42; 95%CI, 0.24-0.74) than pharmacist-directed care (OR, 0.89; 95% CI, 0.68-1.17). CONCLUSIONS: Pharmacist care in the treatment of patients with HF greatly reduces the risk of all-cause and HF hospitalizations. Since hospitalizations associated with HF are a major public health problem, the incorporation of pharmacists into HF care teams should be strongly considered. PMID- 18413551 TI - Computed tomographic colonography to screen for colorectal cancer, extracolonic cancer, and aortic aneurysm: model simulation with cost-effectiveness analysis. AB - BACKGROUND: In addition to detecting colorectal neoplasia, abdominal computed tomography (CT) with colonography technique (CTC) can also detect unsuspected extracolonic cancers and abdominal aortic aneurysms (AAA).The efficacy and cost effectiveness of this combined abdominal CT screening strategy are unknown. METHODS: A computerized Markov model was constructed to simulate the occurrence of colorectal neoplasia, extracolonic malignant neoplasm, and AAA in a hypothetical cohort of 100,000 subjects from the United States who were 50 years of age. Simulated screening with CTC, using a 6-mm polyp size threshold for reporting, was compared with a competing model of optical colonoscopy (OC), both without and with abdominal ultrasonography for AAA detection (OC-US strategy). RESULTS: In the simulated population, CTC was the dominant screening strategy, gaining an additional 1458 and 462 life-years compared with the OC and OC-US strategies and being less costly, with a savings of $266 and $449 per person, respectively. The additional gains for CTC were largely due to a decrease in AAA related deaths, whereas the modeled benefit from extracolonic cancer downstaging was a relatively minor factor. At sensitivity analysis, OC-US became more cost effective only when the CTC sensitivity for large polyps dropped to 61% or when broad variations of costs were simulated, such as an increase in CTC cost from $814 to $1300 or a decrease in OC cost from $1100 to $500. With the OC-US approach, suboptimal compliance had a strong negative influence on efficacy and cost-effectiveness. The estimated mortality from CT-induced cancer was less than estimated colonoscopy-related mortality (8 vs 22 deaths), both of which were minor compared with the positive benefit from screening. CONCLUSION: When detection of extracolonic findings such as AAA and extracolonic cancer are considered in addition to colorectal neoplasia in our model simulation, CT colonography is a dominant screening strategy (ie, more clinically effective and more cost-effective) over both colonoscopy and colonoscopy with 1-time ultrasonography. PMID- 18413552 TI - Length of hospital stay and postdischarge mortality in patients with pulmonary embolism: a statewide perspective. AB - BACKGROUND: The optimal length of stay (LOS) for patients with pulmonary embolism (PE) is unknown. Although reducing LOS is likely to save costs, the effects on patient safety are unclear. We sought to identify patient and hospital factors associated with LOS and assess whether LOS was associated with postdischarge mortality. METHODS: We evaluated patients discharged with a primary diagnosis of PE from 186 acute care hospitals in Pennsylvania (January 2000 through November 2002). We used discrete survival models to examine the association between (1) patient and hospital factors and the time to discharge and (2) LOS and postdischarge mortality within 30 days of presentation, adjusting for patient and hospital factors. RESULTS: Among 15 531 patient discharges with PE, the median LOS was 6 days, and postdischarge mortality rate was 3.3%. In multivariate analysis, patients from Philadelphia were less likely to be discharged on a given day (odds ratio [OR], 0.82; 95% confidence interval [CI], 0.73-0.93), as were black patients (OR, 0.88; 95% CI, 0.82-0.94).The odds of discharge decreased notably with greater patient severity of illness and in patients without private health insurance. Adjusted postdischarge mortality was significantly higher for patients with an LOS of 4 days or less (OR, 1.55; 95% CI, 1.21-2.00) relative to those with an LOS of 5 to 6 days. CONCLUSIONS: Several hospital and patient factors were independently associated with LOS. Patients with a very short LOS had greater postdischarge mortality relative to patients with a typical LOS, suggesting that physicians may inappropriately select patients with PE for early discharge who are at increased risk of complications. PMID- 18413553 TI - Adherence to a DASH-style diet and risk of coronary heart disease and stroke in women. AB - BACKGROUND: The Dietary Approaches to Stop Hypertension (DASH) diet has been shown to lower blood pressure, but little is known about its long-term effect on cardiovascular end points. Our objective was to assess the association between a DASH-style diet adherence score and risk of coronary heart disease (CHD) and stroke in women. METHODS: In this prospective cohort study, diet was assessed 7 times during 24 years of follow-up (1980-2004) with validated food frequency questionnaires. A DASH score based on 8 food and nutrient components (fruits, vegetables, whole grains, nuts and legumes, low-fat dairy, red and processed meats, sweetened beverages, and sodium) was calculated. Lifestyle and medical information was collected biennially with a questionnaire. The Cox proportional hazard model was used to adjust for potential confounders. The study population comprised 88,517 female nurses aged 34 to 59 years without a history of cardiovascular disease or diabetes in 1980. The main outcome measures were the numbers of confirmed incident cases of nonfatal myocardial infarction, CHD death, and stroke. RESULTS: We documented 2129 cases of incident nonfatal myocardial infarction, 976 CHD deaths, and 2317 [corrected] cases of stroke. After adjustment for age, smoking, and other cardiovascular risk factors, the relative risks of CHD across quintiles of the DASH score were 1.0, 0.99, 0.86, 0.87, and 0.76 (95% confidence interval, 0.67-0.85) (P<.001 for trend). The magnitude of risk difference was similar for nonfatal myocardial infarction and fatal CHD. The DASH score was also significantly associated with lower risk of stroke (multivariate relative risks across quintiles of the DASH score were 1.0, 0.92, 0.91, 0.89, and 0.82) (P=.002 for trend). Cross-sectional analysis in a subgroup of women with blood samples showed that the DASH score was significantly associated with lower plasma levels of C-reactive protein (P=.008 for trend) and interleukin 6 (P=.04 for trend). CONCLUSION: Adherence to the DASH-style diet is associated with a lower risk of CHD and stroke among middle-aged women during 24 years of follow-up. PMID- 18413554 TI - Reduction in blood pressure with statins: results from the UCSD Statin Study, a randomized trial. AB - BACKGROUND: Some studies have suggested reductions in blood pressure (BP)with statin treatment, particularly in persons with hypertension. Randomized trial evidence is limited. METHODS: We performed a randomized, double-blind, placebo controlled trial with equal allocation to simvastatin, 20 mg; pravastatin sodium,40 mg; or placebo for 6 months. Nine hundred seventy-three men and women without known cardiovascular disease or diabetes mellitus, with low-density lipoprotein cholesterol screening levels of 115 to 190 mg/dL, had assessment of systolic and diastolic BP (SBP and DBP, respectively). Blood pressure values were compared for placebo vs statins by intention-to-treat (ITT) analysis. Additional analyses were performed that (1) were confined to subjects with neither high baseline BP (SBP>140 mm Hg or DBP>90 mm Hg) nor receiving BP medications, to exclude groups in whom BP medications or medication changes may have influenced results, and (2) separately evaluated simvastatin and pravastatin (vs placebo). The time course of BP changes after statin initiation and the effect of stopping statins on BP were examined. RESULTS: Statins modestly but significantly reduced BP relative to placebo,by 2.2 mm Hg for SBP (P=.02) and 2.4 mm Hg for DBP (P<.001) in ITT analysis. Blood pressure reductions ranged from 2.4 to 2.8 mm Hg for both SBP and DBP with both simvastatin and pravastatin, in those subjects with full follow-up, and without potential for influence by BP medications (ie, neither receiving nor meriting BP medications). CONCLUSIONS: Reductions in SBP and DBP occurred with hydrophilic and lipophilic statins and extended to normotensive subjects. These modest effects may contribute to the reduced risk of stroke and cardiovascular events reported on statins. Trial Registration clinicaltrials.gov Identifier: NCT00330980. PMID- 18413555 TI - Association between cancer risk perception and screening behavior among diverse women. AB - BACKGROUND: We measured the perception of breast, cervical, and colon cancer risks and screening in diverse women to examine the association between risk perception and screening behavior. METHODS: Cross-sectional telephone and in person interviews of women aged 50 to 80 years were conducted in English, Spanish, or Chinese. The women were recruited from primary care practices in San Francisco, California (academic general internal medicine, family medicine, women's health practices, a community-based clinic in Chinatown, and the Community Health Network Clinics, which is affiliated with the San Francisco Department of Public Health), with at least 1 visit within the previous 2 years. Perceived personal risk for each cancer was measured on a word scale (no risk to very high risk) and compared with self-reported screening behavior by ethnicity. RESULTS: A total of 1160 women participated: 338 (29%) were White, 167 (14%) were African American, 239 (21%) were Latina, and 416 (36%)were Asian. The average participant was 61 years old and a high school graduate; 18% had a personal history of cancer, and 42% had a family history of cancer. The perceived lifetime risk of cancer varied by ethnicity. Compared with White women, Latinas had a higher perceived risk for cervical cancer (odds ratio [OR], 2.9; 95% confidence interval [CI], 1.8-4.6) and colon cancer (OR, 3.0; 95% CI, 1.8-5.0) after multivariate adjustment, and Asians had a lower perceived risk for cervical cancer (OR, 0.6; 95% CI, 0.4-0.9) and colon cancer (OR, 0.6; 95% CI, 0.3-0.9). Higher colon cancer risk perception was associated with having undergone colonoscopy within 10 years (OR, 2.8; 95% CI, 1.4-5.4). CONCLUSIONS: Risk perception was significantly associated with colon cancer screening behavior (P=.001). Evaluation of patients' perceived risk of cancer may be useful to clinicians who are recommending screening tests. PMID- 18413556 TI - Loop diuretic use and increased rates of hip bone loss in older men: the Osteoporotic Fractures in Men Study. AB - BACKGROUND: Older adults commonly use loop diuretics, which can increase urinary calcium excretion, leading to potential bone loss. Studies examining the association between loop diuretics and bone mineral density (BMD) are lacking, particularly those involving men. METHODS: In this cohort study, we ascertained medication use (interviewer-administered questionnaire verified with inspection of medication containers) and measured the BMD of the total hip and 2 subregions (by dual-energy x-ray absorptiometry) at baseline and at a second visit an average of 4.6 years later among 3269 men aged 65 years and older. RESULTS: Eighty-four men were categorized as continuous users of loop diuretics, 181 as intermittent users of loop diuretics, and 3004 men as nonusers of loop diuretics. After adjustment for age, baseline BMD, body mass index, weight change from baseline, physical activity,clinic site, perceived health status, cigarette smoking status, diabetes mellitus, chronic obstructive pulmonary disease, congestive heart failure, hypertension, and statin use, the average annual rate of decline in total hip BMD steadily increased from -0.33% (95% confidence interval [CI], -0.36% to -0.31%) for nonusers,to -0.58% (95% CI, -0.69% to 0.47%) for intermittent users, and to -0.78% (95% CI, -0.96% to -0.60%)for continuous users. Findings were similar for change in BMD at the femoral neck and trochanter. CONCLUSIONS: We conclude that loop diuretic use in older men is associated with increased rates of hip bone loss. These results suggest that the potential for bone loss should be considered when loop diuretics are prescribed to older patients in clinical practice. PMID- 18413557 TI - Clinical uncertainty, diagnostic accuracy, and outcomes in emergency department patients presenting with dyspnea. AB - BACKGROUND: Dyspnea is a common complaint in the emergency department (ED) and may be a diagnostic challenge. We hypothesized that diagnostic uncertainty in this setting is associated with adverse outcomes, and amino-terminal pro-B-type natriuretic peptide (NT-proBNP) testing would improve diagnostic accuracy and reduce diagnostic uncertainty. METHODS: A total of 592 dyspneic patients were evaluated from the ProBNP Investigation of Dyspnea in the Emergency Department (PRIDE) study. Managing physicians were asked to provide estimates from 0% to 100%of the likelihood of acutely destabilized heart failure (ADHF). A certainty estimate of either 20% or lower or 80% or higher was classified as clinical certainty, while estimates between 21% and 79% were defined as clinical uncertainty. Associations between clinical uncertainty,hospital length of stay, morbidity, and mortality were examined. The diagnostic value of clinical judgment vs NT-proBNP measurement was compared across categories of clinical certainty. RESULTS: Clinical uncertainty was present in 185 patients (31%), 103 (56%) of whom had ADHF. Patients judged with clinical uncertainty had longer hospital length of stay and increased morbidity and mortality,especially those with ADHF. Receiver operating characteristic analysis of clinical judgment yielded an area under the curve (AUC) of 0.88 in the clinical certainty group and 0.76 in the uncertainty group (P<.001); NT-proBNP testing alone in these same groups had AUCs of 0.96 and 0.91, respectively. The combination of clinical judgment with NT proBNP testing yielded improvements in AUC. CONCLUSIONS: Among dyspneic patients in the ED, clinical uncertainty is associated with increased morbidity and mortality, especially in those with ADHF.The addition of NT-proBNP testing to clinical judgment may reduce diagnostic uncertainty in this setting. PMID- 18413558 TI - Invasive pneumococcal disease among White Mountain Apache adults, 1991-2005. AB - BACKGROUND: Certain Native American populations have high rates of invasive pneumococcal disease (IPD). We aimed to determine the disease spectrum and risk factors of White Mountain Apache adults (age, >or=18 years) with IPD and the use and effectiveness of pneumococcal polysaccharide vaccine (PPV) in this population. METHODS: We conducted active surveillance for IPD between 1991 and 2005. Medical records were reviewed, and isolates were serotyped. Vaccine use was assessed in 2004-2005 among White Mountain Apache adults with an indication for pneumococcal vaccination. The effectiveness of PPV was determined through an indirect cohort method. RESULTS: Among the 115 IPD cases (in 109 persons), the mean age was 43 years; 62% were male; 91% had risk factors, and alcoholism predominated (73%). Alcoholic patients were younger (mean age, 40.1 years; P<.001) and more often male (70%; P=.001) compared with nonalcoholic patients. The case fatality rate was 15%; all deaths occurred among those with risk factors. Only age 65 years or older was associated with increased risk of death. Of 447 White Mountain Apache persons at high risk, 76% had received PPV. Vaccination rates were highest among subjects with pulmonary disease (95%) and diabetes (89%) and lowest among those aged 50 to 64 years (40%). Of the 115 IPD cases for which serotypes were available, 77% were due to serotypes contained in PPV. The effectiveness of PPV against serotype-specific IPD, as measured by the indirect cohort analysis of IPD cases, was 68% (95% confidence interval, 3%-90%). CONCLUSIONS: Among White Mountain Apache adults with IPD, alcoholism is common and contributes to the younger age and male predominance of cases. Pneumococcal vaccination rates are high, and there is suggestive evidence of the effectiveness of PPV in this population. Additional preventive strategies, including risk factor modification and vaccination of younger high-risk adults, should be pursued. PMID- 18413559 TI - Incidental findings on cardiac multidetector row computed tomography among healthy older adults: prevalence and clinical correlates. AB - BACKGROUND: With the widespread use of cardiac multidetector row computed tomography (MDCT), the issue of incidental findings is receiving increasing attention. Our objectives were to evaluate the prevalence of incidental findings discovered during cardiac MDCT scanning and to identify clinical variables associated with incidental findings. METHODS: This cross-sectional analysis involved a population-based sample recruited from an integrated health care delivery system in Northern California as part of the Atherosclerotic Disease, Vascular Function and Genetic Epidemiology (ADVANCE) Study. Healthy men and women aged 60 to 69 years without diagnosed cardiovascular disease underwent cardiac MDCT for the detection and quantification of coronary artery calcification. The images were prospectively evaluated for incidental findings. RESULTS: A total of 459 participants underwent MDCT scanning, and the overall prevalence of any incidental finding was 41%. Of the 459 participants, 105 (23%) had at least 1 incidental finding that was recommended for clinical or radiological follow-up examination, the most common of which was single or multiple pulmonary nodules (18%). Participants with and without incidental findings had comparable baseline demographics and selected clinical variables, although there were significantly fewer men and a significantly lower prevalence of the metabolic syndrome in those with incidental findings. CONCLUSIONS: Incidental findings, especially pulmonary nodules, are common in cardiac MDCT performed to assess coronary artery calcification in older healthy adults. The net risks and benefits of looking for noncardiac abnormalities during cardiac MDCT should be rigorously evaluated. PMID- 18413560 TI - Association of beta1-adrenergic receptor genetic polymorphism with mental stress induced myocardial ischemia in patients with coronary artery disease. AB - BACKGROUND: Mental stress is associated with sympathetic adrenergic stimulation and concomitant increases in blood pressure and heart rate. Heritable individual differences in cardiovascular functional response to mental stress may arise from genetic variations in adrenergic receptors, which might produce excessive hemodynamic response to mental stress or create other conditions favoring the development of myocardial ischemia. METHODS: We examined the relationship between hemodynamic response to mental stress and mental stress-induced myocardial ischemia (MSIMI) and 5 common functional polymorphisms of beta1-adrenergic receptors (ADRB1 [OMIM 109630, accession No. 153]) and beta2-adrenergic receptors (ADRB2 [OMIM 109690, accession No. 154]). Participants were 148 patients (45 female [30.4%]) with a documented history of coronary artery disease and a mean (SD) age of 64 (9) years. Patients were enrolled between December 9, 2004, and February 21, 2007. Mental stress was induced via a public-speaking task. Rest and stress myocardial perfusion imaging was performed. Blood samples were collected and genotyped for 5 common functional polymorphisms of ADRB1 (codons 49 and 389) and ADRB2 (codons 16 and 27 and nucleotide 523). The main outcome measures were hemodynamic and myocardial ischemic responses to mental stress. Mental stress induced myocardial ischemia was defined as new or worsening perfusion defects during mental stress with a summed (stress to rest) difference score of at least 3. RESULTS: A statistically significant difference was noted in the prevalences of MSIMI between genotype groups for codon 49 of ADRB1. Mental stress-induced myocardial ischemia occurred 3 times more frequently among patients homozygous for the Ser49 allele (31 of 104 patients [29.8%]) compared with 4 of 39 patients (10.3%) among the Gly49 allele carriers (P=.02). The adjusted odds ratio for the effect of genotype (Ser/Ser vs Gly carriers) on MSIMI was 3.9 (95% confidence interval, 1.2-12.5) (P=.02). CONCLUSIONS: Our findings indicate an association between a common genetic variation in ADRB1 and myocardial ischemic response to mental stress in patients with coronary artery disease. This polymorphic genetic marker may help identify patients at increased risk for mental stress-induced adverse outcomes. PMID- 18413561 TI - Vitamin D as an analgesic for patients with type 2 diabetes and neuropathic pain. PMID- 18413562 TI - Persistent Staphylococcus aureus bacteremia. PMID- 18413563 TI - Clinical trials of antioxidant supplementation in the prevention of cardiovascular events. PMID- 18413564 TI - Hunting for genes and mutations: it's worth remembering the basics. PMID- 18413565 TI - Complicated autosomal recessive hereditary spastic paraplegia: a complex picture is emerging. PMID- 18413566 TI - Cognitive impairments in multiple system atrophy: MSA-C vs MSA-P. AB - OBJECTIVE: We evaluated comprehensive neuropsychological tests and regional brain blood flow to compare cognitive dysfunction between two types of multiple system atrophy: predominant cerebellar ataxia (MSA-C) and predominant parkinsonism (MSA P). METHODS: Twenty-one patients with MSA-C, 14 patients with MSA-P, and 21 age- and education-matched control subjects were subjected to neuropsychological tests and SPECT. The neuropsychological tests examined general cognition, verbal and visual memory, working memory, visuospatial and constructional ability, language, executive function, depression, and anxiety, while SPECT analysis examined brain perfusion. RESULTS: Patients with MSA-P showed severe involvement of visuospatial and constructional function, verbal fluency, and executive function compared with control subjects. Patients with MSA-C showed involvement only in visuospatial and constructional function compared with control subjects and a milder degree of involvement compared with patients with MSA-P. Patients with MSA-P tended toward a wide and severe impairment in cognitive function compared with patients with MSA-C. In addition, neuropsychological impairment in patients with MSA-P was significantly correlated with a decrease in prefrontal perfusion. This significant relation was not correlated to other factors such as age, education, and severity of cerebellar ataxia and parkinsonism, which are relevant factors associated with cognitive performance. CONCLUSIONS: Patients with multiple system atrophy-parkinsonism show more severe and more widespread cognitive dysfunctions than patients with multiple system atrophy-cerebellar ataxia. Our results also indicate that cognitive dysfunction in patients with multiple system atrophy parkinsonism may be associated with prefrontal involvement. PMID- 18413567 TI - Parkinson disease and risk of mortality: a prospective comorbidity-matched cohort study. AB - OBJECTIVE: To evaluate the association between Parkinson disease (PD) and mortality after adjustment for comorbidities. METHODS: We conducted a matched cohort analysis among 22,071 participants in the Physicians' Health Study. Five hundred sixty incident PD cases were identified by self-report. We used a modified Charlson Comorbidity Index to calculate a comorbidity score. Each PD case was matched by age to a comparator who was alive and had an identical comorbidity score at the time of PD diagnosis of the case. Both cohorts were followed for all-cause mortality. We used proportional hazards models to calculate hazard ratios (HRs) for mortality. RESULTS: A total of 330 participants died over a median follow-up of 5.8 years, 200 (35.7%) in the PD group and 130 (23.2%) in the reference group. After adjustment for smoking and age at PD onset, the HR for mortality was 2.32 (95% CI 1.85-2.92). The mortality risk remained significant with increasing age at onset, even in those aged >or=80 years (HR = 2.10; 95% CI 1.44-3.00). The increased risk was apparent for short PD duration (<2 years) and remained stable with increasing duration. We found no different risk of mortality associated with PD according to smoking status. CONCLUSIONS: In this large prospective cohort of men and after matching on comorbidities, we found that Parkinson disease patients had an increased risk of all-cause mortality. Mortality was increased regardless of disease duration, did not diminish with increasing age at onset, and was not influenced by smoking status. PMID- 18413568 TI - Effects of subthalamic nucleus stimulation and levodopa on freezing of gait in Parkinson disease. AB - OBJECTIVE: We studied the effects of subthalamic nucleus (STN) stimulation vs levodopa on freezing of gait (FOG) and gait impairments in a large consecutive series of patients with Parkinson disease with bilateral STN stimulation. METHODS: One hundred twenty-three patients performed the Stand Walk Sit Test before and 1 year after surgery both off and on levodopa and off and on stimulation. RESULTS: Before surgery, 25 patients displayed FOG episodes and 48 were unable to complete the Stand Walk Sit Test when off levodopa. Both symptoms were alleviated by levodopa. After surgery, STN stimulation reproduced the improvement induced by levodopa before surgery in all but two patients with FOG and five others unable to walk. In 11 patients, FOG or inability to perform the test first occurred after surgery. In all patients but those experiencing FOG during the Stand Walk Sit Test before surgery, the benefit of STN stimulation did not reach that of levodopa before surgery. In patients with FOG before surgery, the effect of STN stimulation did not differ from that of levodopa either before or after surgery. CONCLUSIONS: Overall, subthalamic nucleus stimulation improved levodopa-responsive freezing of gait in most patients, although it was not always as effective as levodopa to improve gait impairments. In addition, surgery can induce gait problems in some patients. PMID- 18413569 TI - Subthalamic nucleus stimulation modulates audiospinal reactions in Parkinson disease. AB - BACKGROUND: Axial symptoms of Parkinson disease (PD) may result from dysfunctional basal ganglia-brainstem connections. In this study, we assessed whether modulation of basal ganglia activity by high-frequency stimulation of the subthalamic nucleus (STN-HFS) in PD had an impact on the brainstem-controlled startle system. METHODS: We assessed auditory startle responses (recorded from right orbicularis oculi, masseter, sternocleidomastoid, biceps brachii, and soleus muscle) and audiospinal facilitation (startle conditioned soleus H reflexes at interstimulus intervals of 0-250 msec) in 24 patients with PD with chronically implanted, bilateral STN electrodes in the stimulation on (STIM ON) and off condition (STIM OFF) and 20 healthy controls. RESULTS: The mixed linear analysis of variance model revealed a significant effect for the startle onset latency in the orbicularis oculi muscle for the factors GROUP (patients with PD vs controls; p < 0.0001, F = 44.66) and STIM (nested within GROUP) (p = 0.0034, F = 8.79). Audiospinal facilitation was modulated by STN-HFS as shown by highly significant effects for STIM [GROUP] (p < 0.0001, F = 15.9), ISI [GROUP] (p < 0.0001, F = 3.5), and the interaction of ISI x STIM [GROUP] (p = 0.0085, F = 2.65) in the mixed linear model. CONCLUSION: High-frequency stimulation of the subthalamic nucleus alters the excitability of the brainstem startle system in Parkinson disease, most likely by releasing the reticular motor system from abnormal descending input of the basal ganglia via pallidotegmental pathways. PMID- 18413570 TI - Pathologic findings in prospectively ascertained essential tremor subjects. AB - OBJECTIVE: To assess pathologic changes in prospectively characterized subjects with essential tremor (ET). METHODS: Subjects enrolled in the Sun Health Research Institute Brain and Body Donation Program were examined annually by a movement disorders neurologist, and semiannually by a behavioral neurologist and neuropsychologist. Twenty-four subjects without a prior diagnosis of dementia or other major movement disorder met clinical criteria for ET and came to autopsy. Subjects with mild cognitive impairment (n = 3) were included. These subjects were compared with 21 controls. Brains were examined postmortem according to standardized protocols for assessment of age-related changes and specific pathologic conditions (e.g., Parkinson disease, Alzheimer disease). RESULTS: Subjects had a mean age of 86.2 years and a mean duration of tremor of 11.1 years. Seven subjects had evidence for cerebellar pathology (Purkinje cell loss, cerebellar cortical sclerosis, and proliferation of Bergmann glia). Pigmented neurons were qualitatively depleted in the locus ceruleus in eight subjects and in the substantia nigra in five subjects. Of these, three had Lewy bodies, one subject had brainstem predominant disease, and two had limbic stage. Three subjects had a nonspecific cerebral tauopathy and another met pathologic criteria for progressive supranuclear palsy. However, when compared with controls, only changes in the locus ceruleus and gliosis of the cerebellum remained significant findings. CONCLUSIONS: This study supports previous findings of heterogenous pathology in essential tremor (ET). There is an increased frequency of cerebellar gliosis and locus ceruleus depletion. We did not find an increased incidence of Lewy bodies in subjects with ET. PMID- 18413571 TI - Functional imaging in Parkinson disease. AB - OBJECTIVE: Functional imaging techniques represent useful tools to assess in vivo the neurochemical alterations and functional connectivity in Parkinson disease (PD). Here, the authors review the various approaches and potential application of these imaging techniques to the study of PD. METHOD: Radiotracer imaging using dopaminergic markers facilitates assessment of pre- and postsynaptic nigrostriatal integrity, while imaging with other appropriate radiotracers explores nondopaminergic neurotransmitter function, local metabolism, blood flow, and mechanisms potentially related to disease progression and pathogenesis. Activation studies using functional MRI detect blood oxygen level dependent signal, as an indirect marker of neuronal activity. RESULT: Functional imaging techniques have been applied to infer the potential role of inflammation and other factors in etiopathogenesis as well as to study compensatory and regulatory mechanisms in early PD and subclinical disease in genetic forms of PD. Imaging studies also help to understand the neurobiological basis of motor and nonmotor complications. Recent reports suggest a role for striatal dopaminergic transmission in modulating neurobehavioral processes including the placebo effect in PD. Although functional imaging has been employed to monitor disease progression, the discordance between clinical outcome and imaging measures after therapeutic interventions precludes their use as surrogate end points in clinical trials. Beyond these limitations and potential challenges, imaging techniques continue to find wide application in the study of PD. CONCLUSION: Functional imaging can provide meaningful insights into mechanisms underlying various aspects of motor and nonmotor dysfunction in Parkinson disease and the role of striatal dopaminergic transmission in behavioral processes beyond motor control. These modalities hold promise to study the preclinical phase and to elucidate further the benefits and complications of surgical interventions and the utility of neuroprotective strategies. PMID- 18413572 TI - Postmortem proof of effectiveness of zona incerta stimulation in Parkinson disease. PMID- 18413573 TI - PARK9-linked parkinsonism in eastern Asia: mutation detection in ATP13A2 and clinical phenotype. PMID- 18413574 TI - Neuroferritinopathy in a Japanese family with a duplication in the ferritin light chain gene. PMID- 18413576 TI - Graves disease and isolated orthostatic tremor. PMID- 18413577 TI - Psychogenic tremor occurring after deep brain stimulation surgery for essential tremor. PMID- 18413575 TI - Lack of association between SLITRK1var321 and Tourette syndrome in a large family based sample. PMID- 18413578 TI - Thalamic Vo-complex vs pallidal deep brain stimulation for focal hand dystonia. PMID- 18413579 TI - Novel ATP1A3 mutation in a sporadic RDP patient with minimal benefit from deep brain stimulation. PMID- 18413581 TI - Emerging subspecialties: neurorehabilitation: training neurologists to retrain the brain. PMID- 18413582 TI - Right brain: the disentanglement. PMID- 18413584 TI - Calcitonin gene-related peptide (CGRP) antagonists and migraine: is this a new era? PMID- 18413585 TI - Will my Rett syndrome patient walk, talk, and use her hands? PMID- 18413586 TI - High pulse pressure protects against headache: prospective and cross-sectional data (HUNT study). AB - BACKGROUND: Many antihypertensive drugs are also used as migraine prophylactics, but the relationship between blood pressure and headache is not been well understood. The objective of the present study was to explore the association between blood pressure and headache prevalence, and the effect of antihypertensive medication on this relationship, using both cross-sectional and prospective data from a large population. METHODS: We used data from two large epidemiologic studies, the Nord-Trondelag Health Survey 1984-1986 (HUNT-1) and 1995-97 (HUNT-2), to evaluate the association between blood pressure (systolic, diastolic, mean arterial, and pulse pressure) and migraine and nonmigrainous headache. RESULTS: Increasing systolic blood pressure was associated with decreasing prevalence of having nonmigrainous headache or migraine. The most consistent and robust finding, however, was that increasing pulse pressure was linked to decreased prevalence of both nonmigrainous headache and migraine, evident for both sexes in the prospective and cross-sectional analyses. In subjects using antihypertensive medication, this finding was less clear. CONCLUSION: Both increased systolic blood pressure and pulse pressure are related to arterial stiffness and may decrease headache prevalence through modulation of the baroreflex arch, which in turn generates hypoalgesia. This is due to a phenomenon called hypertension-associated hypalgesia. Stimulation of the baroreflex arch in response to increased blood pressure is assumed to inhibit pain transmission at both spinal and supraspinal levels, possibly because of an interaction of the centers modulating nociception and cardiovascular reflexes in the brainstem. PMID- 18413587 TI - Validity of the ID-Migraine screener in the workplace. AB - OBJECTIVE: The impact of migraine on physical, social, and emotional performance is considerable, yet it remains an underdiagnosed disorder. ID-Migraine is a validated migraine-screening tool developed to facilitate diagnosis. This study evaluated the validity and use of the Turkish version of the ID-Migraine screener (ID-Ms) in the workplace, and measured the impact of headache on disability, productivity, and quality of life among the workforce. METHODS: A total of 465 employees from four companies were interviewed for screening with the ID-Ms. Subjects were included in the study if they reported two or more headaches in the past 3 months and gave a positive answer to one of the two ID-Ms prescreening questions. Eligible subjects completed the ID-Ms, the Migraine Disability Assessment Questionnaire, and the Medical Outcomes Study 36-Item Short Form Health Survey. Subjects were then evaluated for confirmation of their diagnosis according to the International Classification of Headache Disorders, 2nd edition (ICHD-2) criteria. RESULTS: A total of 227 subjects (mean age 31.9 +/- 5.9 years; 65.6% women) completed the study. Migraine was diagnosed in 106 of the 227 subjects (46.7%) according to the ID-Ms and in 117 of the 227 subjects (51.5%) according to ICHD-2 criteria. The sensitivity of the ID-Ms was 70.9%, specificity was 79.1% and Cohen kappa value was 0.50. Workdays lost over the previous 3 months due to headache amounted to 8.7 +/- 9.5 days for migraine-positive and 4.9 +/- 6.6 days for migraine-negative subjects. CONCLUSION: The Turkish version of the ID-Migraine screener is a valid tool for identifying subjects with migraine in the workplace. PMID- 18413588 TI - Sexual function in men with epilepsy: how important is testosterone? AB - OBJECTIVE: To ascertain the effects on sexual function of men with epilepsy (MWE) of testosterone levels and indices of anxiety and depression. METHODS: Sixty MWE taking one antiepileptic drug only (AED), with no comedication, were compared with 60 control men. Total testosterone (TT), free testosterone (FT), bioactive testosterone (BAT), dehydroepiandrosterone sulfate (DHEAS), androstenedione, and sex hormone-binding globulin (SHBG) were measured. Each man also completed validated questionnaires exploring sexual desire (Sexual Desire Inventory [SDI]), sexual response (Sexual Response Inventory [SRI]), erectile function (Sexual Self Efficacy Scale [SSES]), and anxiety and depression (Hospital Anxiety and Depression Scale). RESULTS: MWE reported lower levels of sexual desire and lower erectile function compared with controls. They had significantly higher levels of anxiety, depression, and psychological distress. MWE had significantly higher SHBG levels and significantly lower DHEAS. There were no significant differences between the groups' TT, FT, or BAT levels. BAT levels were significantly lower in men taking enzyme-inducing AEDs than in those taking non-enzyme-inducing AEDs. Visual inspection of TT and BAT levels showed that the majority of MWE and controls had TT and BAT levels above the "androgen threshold" levels of 12 nmol/L TT or 3.8 nmol/L BAT considered necessary for normal sexual function. There was a significant correlation (Spearman rank and simple linear regression) between sexual function and indices of anxiety and depression. There was no significant relationship between SDI and SSES and TT, FT, or BAT (Spearman rank correlation). CONCLUSIONS: Concentrating on hormone levels alone as an explanation of sexual dysfunction in epilepsy represents an overly simplistic approach to the problem. Future studies should include measures of quality of life, anxiety, and depression. PMID- 18413589 TI - N-acetylaspartate and N-acetylaspartylglutamate: neurobiology and clinical significance. PMID- 18413590 TI - Extension of the clinical spectrum of Danon disease. PMID- 18413591 TI - Mitochondrial diseases associated with cerebral folate deficiency. PMID- 18413592 TI - Extracephalic cluster (cluster sine headache). PMID- 18413593 TI - Imaging of Bing-Neel syndrome. PMID- 18413594 TI - Re: What is next in ALS clinical trials? PMID- 18413595 TI - Re: What is next in ALS clinical trials? PMID- 18413596 TI - Understanding epilepsy through network modeling. PMID- 18413597 TI - Adenovirus E1A targets p400 to induce the cellular oncoprotein Myc. AB - Adenovirus E1A drives oncogenesis by targeting key regulatory pathways that are critical for cellular growth control. The interaction of E1A with p400 is essential for many E1A activities, but the downstream target of this interaction is unknown. Here, we present evidence that the oncoprotein transcription factor Myc is the target of this interaction. We show that E1A stabilizes Myc protein via p400 and promotes the coassociation of Myc and p400 at Myc target genes, leading to their transcriptional induction. We also show that E1A requires Myc for its ability to activate Myc-dependent gene expression and induce apoptosis, and that forced expression of Myc is sufficient to rescue the activity of an E1A mutant defective in p400 binding. Together, these findings establish that Myc, via p400, is an essential downstream target of E1A. PMID- 18413598 TI - Adipogenic capacity and the susceptibility to type 2 diabetes and metabolic syndrome. AB - To determine whether adipocyte storage capacity influences the onset and severity of type 2 diabetes and other components of the metabolic syndrome, we made normal and db/db mice resistant to obesity by overexpressing leptin receptor-b on the aP2-Lepr-b promoter. On a 4% diet, these mice have no phenotype, but on a 60% fat diet, they resist diet-induced obesity because constitutive adipocyte-specific overexpression of Lepr-b prevents obesity via the antilipogenic autocrine/paracrine action of leptin on adipocytes. After 8 months on the same 60% fat diet, body fat of transgenic mice was 70% below WT controls. Cardiac and liver fat was elevated in the transgenics, and their hyperinsulinemia was more marked, suggesting greater insulin resistance. The aP2-Lepr-b transgene also prevented obesity in db/db mice; at 10 weeks of age their body fat was half that of the db/db mice. This lack of obesity was attributable to reduced expression of sterol regulatory element binding protein-1c and its target lipogenic enzymes in adipose tissue and a 6-fold increase in Pref-1 mRNA. Severe diabetes was present in transgenics at 4 weeks of age, 10 weeks before db/db controls. Echocardiographic evidence of cardiomyopathy appeared at 10 weeks, weeks before the db/db mice. Histologically, loss of beta cells and myocardial fibrosis was present in the transgenic group at least 6 weeks before the db/db mice. These results suggest that the expression level of genes that regulate the adipogenic response to overnutrition profoundly influences the age of onset and severity of diet-induced type 2 diabetes and co-morbidities. PMID- 18413599 TI - Global metabolic responses of mice to Trypanosoma brucei brucei infection. AB - Human African trypanosomiasis (HAT) is transmitted by tsetse flies and, if untreated, is fatal. Treatment depends on infection stage, and early diagnosis is crucial for effective disease management. The systemic host biochemical changes induced by HAT that enable biomarker discovery or relate to therapeutic outcome are largely unknown. We have characterized the multivariate temporal responses of mice to Trypanosoma brucei brucei infection, using (1)H nuclear magnetic resonance (NMR) spectroscopic metabolic phenotyping of urine and plasma. Marked alterations in plasma metabolic profiles were detected already 1 day postinfection. Elevated plasma concentrations of lactate, branched chain amino acids, and acetylglycoprotein fragments were noted. T. brucei brucei-infected mice also had an imbalance of plasma alanine and valine, consistent with differential gluconeogenesis (parasite)-ketogenesis (host) pathway counterflux, involving stimulated host glycolysis, ketogenesis, and enhanced lipid oxidation in the host. Histopathologic evidence of T. brucei brucei-induced extramedullary hepatic hemopoiesis, renal interstitial nephritis, and a provoked inflammatory response was also noted. Metabolic disturbance of gut microbiotal activity was associated with infection, as indicated by changes in the urinary concentrations of the microbial co-metabolites, including hippurate. Concluding, parasite infection results in multiple systemic biochemical effects in the host and disturbance of the symbiotic gut microbial metabolic interactions. Investigation of these transgenomic metabolic alterations may underpin the development of new diagnostic criteria and metrics of therapeutic efficacy. PMID- 18413600 TI - Identification and evolution of fungal mitochondrial tyrosyl-tRNA synthetases with group I intron splicing activity. AB - The bifunctional Neurospora crassa mitochondrial tyrosyl-tRNA synthetase (CYT-18 protein) both aminoacylates mitochondrial tRNA(Tyr) and acts as a structure stabilizing splicing cofactor for group I introns. Previous studies showed that CYT-18 has distinct tRNA(Tyr) and group I intron-binding sites, with the latter formed by three small "insertions" in the nucleotide-binding fold and other structural adaptations compared with nonsplicing bacterial tyrosyl-tRNA synthetases. Here, analysis of genomic sequences shows that mitochondrial tyrosyl tRNA synthetases with structural adaptations similar to CYT-18's are uniquely characteristic of fungi belonging to the subphylum Pezizomycotina, and biochemical assays confirm group I intron splicing activity for the enzymes from several of these organisms, including Aspergillus nidulans and the human pathogens Coccidioides posadasii and Histoplasma capsulatum. By combining multiple sequence alignments with a previously determined cocrystal structure of a CYT-18/group I intron RNA complex, we identify conserved features of the Pezizomycotina enzymes related to group I intron and tRNA interactions. Our results suggest that mitochondrial tyrosyl-tRNA synthetases with group I intron splicing activity evolved during or after the divergence of the fungal subphyla Pezizomycotina and Saccharomycotina by a mechanism involving the concerted differentiation of preexisting protein loop regions. The unique group I intron splicing activity of these fungal enzymes may provide a new target for antifungal drugs. PMID- 18413601 TI - TNF receptor I sensitizes neurons to erythropoietin- and VEGF-mediated neuroprotection after ischemic and excitotoxic injury. AB - CNS neurons use robust cytoprotective mechanisms to ensure survival and functioning under conditions of injury. These involve pathways induced by endogenous neuroprotective cytokines such as erythropoietin (EPO). Recently, in contrast to its well known deleterious roles, TNF has also been shown to exhibit neuroprotective properties. In the present study, we investigated the molecular mechanisms by which TNF receptor (TNFR)I mediates neuroprotection by comparing the gene expression profiles of lesioned cortex from WT and TNFRI KO mice after permanent middle cerebral artery occlusion. Several known neuroprotective molecules were identified as TNFRI targets, notably members of the Bcl-2 family, DNA repair machinery and cell cycle, developmental, and differentiation factors, neurotransmitters and growth factors, as well as their receptors, including EPO receptor (EPOR), VEGF, colony-stimulating factor receptor 1, insulin-like growth factor (IGF), and nerve growth factor (NGF). Further analysis showed that induction of EPOR and VEGF expression in primary cortical neurons after glucose deprivation (GD) largely depended on TNFRI and was further up-regulated by TNF. Also, EPO- and VEGF-induced neuroprotection against GD, oxygen-glucose deprivation, and NMDA excitotoxicity depended significantly on TNFRI presence. Finally, EPO prevented neuronal damage induced by kainic acid in WT but not TNFRI KO mice. Our results identify cross-talk between tissue protective cytokines, specifically that TNFRI is necessary for constitutive and GD-induced expression of EPOR and VEGF and for EPO-mediated neuroprotection. PMID- 18413602 TI - Blood oxygenation level-dependent contrast response functions identify mechanisms of covert attention in early visual areas. AB - Covert attention can lead to improved performance in perceptual tasks. The neural and functional mechanisms of covert attention are still under investigation. Using both rapid event-related and mixed designs, we measured the blood oxygenation level-dependent functional MRI contrast response functions over the full range of contrast (0-100%) in the retinotopically defined early visual areas (V1, V2, V3, V3A, and V4) in humans. Covert attention increased both the baseline activities and contrast gains in the five cortical areas. The effect on baseline can be decomposed into a transient trial-by-trial component and a component across an entire attention block. On average, increase in contrast gain accounted for approximately 88.0%, 28.5%, 12.7%, 35.9%, and 25.2% of the trial-by-trial effects of attention in the five areas, respectively, and 22.2%, 12.8%, 7.4%, 19.7%, and 17.3% of the total effects of attention in those areas, consistent with single-unit findings in V4 and MT. The results provide strong evidence for a stimulus enhancement mechanism of attention as demonstrated in various behavioral studies. PMID- 18413603 TI - Combinatorial antibody libraries from survivors of the Turkish H5N1 avian influenza outbreak reveal virus neutralization strategies. AB - The widespread incidence of H5N1 influenza viruses in bird populations poses risks to human health. Although the virus has not yet adapted for facile transmission between humans, it can cause severe disease and often death. Here we report the generation of combinatorial antibody libraries from the bone marrow of five survivors of the recent H5N1 avian influenza outbreak in Turkey. To date, these libraries have yielded >300 unique antibodies against H5N1 viral antigens. Among these antibodies, we have identified several broadly reactive neutralizing antibodies that could be used for passive immunization against H5N1 virus or as guides for vaccine design. The large number of antibodies obtained from these survivors provide a detailed immunochemical analysis of individual human solutions to virus neutralization in the setting of an actual virulent influenza outbreak. Remarkably, three of these antibodies neutralized both H1 and H5 subtype influenza viruses. PMID- 18413604 TI - Transfer of a large gene regulatory apparatus to a new developmental address in echinoid evolution. AB - Of the five echinoderm classes, only the modern sea urchins (euechinoids) generate a precociously specified embryonic micromere lineage that ingresses before gastrulation and then secretes the biomineral embryonic skeleton. The gene regulatory network (GRN) underlying the specification and differentiation of this lineage is now known. Many of the same differentiation genes as are used in the biomineralization of the embryo skeleton are also used to make the similar biomineral of the spines and test plates of the adult body. Here, we determine the components of the regulatory state upstream of these differentiation genes that are shared between embryonic and adult skeletogenesis. An abrupt "break point" in the micromere GRN is thus revealed, on one side of which most of the regulatory genes are used in both, and on the other side of which the regulatory apparatus is entirely micromere-specific. This reveals the specific linkages of the micromere GRN forged in the evolutionary process by which the skeletogenic gene batteries were caused to be activated in the embryonic micromere lineage. We also show, by comparison with adult skeletogenesis in the sea star, a distant echinoderm outgroup, that the regulatory apparatus responsible for driving the skeletogenic differentiation gene batteries is an ancient pleisiomorphic aspect of the echinoderm-specific regulatory heritage. PMID- 18413605 TI - Stable isotope evidence for an amphibious phase in early proboscidean evolution. AB - The order Proboscidea includes extant elephants and their extinct relatives and is closely related to the aquatic sirenians (manatees and dugongs) and terrestrial hyracoids (hyraxes). Some analyses of embryological, morphological, and paleontological data suggest that proboscideans and sirenians shared an aquatic or semiaquatic common ancestor, but independent tests of this hypothesis have proven elusive. Here we test the hypothesis of an aquatic ancestry for advanced proboscideans by measuring delta(18)O in tooth enamel of two late Eocene proboscidean genera, Barytherium and Moeritherium, which are sister taxa of Oligocene-to-Recent proboscideans. The combination of low delta(18)O values and low delta(18)O standard deviations in Barytherium and Moeritherium matches the isotopic pattern seen in aquatic and semiaquatic mammals, and differs from that of terrestrial mammals. delta(13)C values of these early proboscideans suggest that both genera are likely to have consumed freshwater plants, although a component of C(3) terrestrial vegetation cannot be ruled out. The simplest explanation for the combined evidence from isotopes, dental functional morphology, and depositional environments is that Barytherium and Moeritherium were at least semiaquatic and lived in freshwater swamp or riverine environments, where they grazed on freshwater vegetation. These results lend new support to the hypothesis that Oligocene-to-Recent proboscideans are derived from amphibious ancestors. PMID- 18413606 TI - Protection against heterologous human papillomavirus challenge by a synthetic lipopeptide vaccine containing a broadly cross-neutralizing epitope of L2. AB - Persistent infection with the high-risk subset of genitotropic human papillomavirus (HPV) genotypes is a necessary cause of cervical cancer. Given the global burden of cervical cancer, a low-cost, broadly protective vaccine is needed. RG-1 is a cross-neutralizing and protective monoclonal antibody that recognizes residues 17-36 of HPV16 minor capsid protein L2. Because this epitope is highly conserved in divergent HPV types, we determined whether vaccination with HPV16 L2 17-36 peptide is broadly protective. The peptide was administered to BALB/c mice three times at monthly intervals, either alone or in the context of a synthetic lipopeptide vaccine candidate (P25-P2C-HPV) produced by linkage of the HPV peptide with a broadly recognized T helper epitope (P25) and the Toll like receptor-2 (TLR2) ligand dipalmitoyl-S-glyceryl cysteine (P2C). In contrast to vaccination with the L2 17-36 peptide or P25-P2C alone, a potent L2-specific antibody response was generated to the P25-P2C-HPV lipopeptide when delivered either s.c. or intranasally. Sera from mice vaccinated with the P25-P2C-HPV lipopeptide neutralized not only HPV16 pseudovirions but also other evolutionarily divergent oncogenic genital (HPV18, HPV45) and cutaneous (HPV5, BPV1) types. The L2-specific antibody response depended on MHC class II, CD40, and MyD88 signaling. Additionally, vaccination with the P25-P2C-HPV lipopeptide protected mice from homologous challenge with HPV16 pseudovirions at cutaneous and genital sites and heterologous challenge with HPV45 pseudovirions. If provided in the appropriate context, therefore, HPV16 L2 17-36 might be used in a totally synthetic cross-protective HPV vaccine. PMID- 18413607 TI - Activation of the lifespan regulator p66Shc through reversible disulfide bond formation. AB - Cell fate and organismal lifespan are controlled by a complex signaling network whose dysfunction can cause a variety of aging-related diseases. An important protection against these failures is cellular apoptosis, which can be induced by p66(Shc) in response to cellular stress. The precise mechanisms of p66(Shc) action and regulation and the function of the p66(Shc)-specific N terminus remain to be identified. Here, we show that the p66(Shc) N terminus forms a redox module responsible for apoptosis initiation, and that this module can be activated through reversible tetramerization by forming two disulfide bonds. Glutathione and thioredoxins can reduce and inactivate p66(Shc), resulting in a thiol-based redox sensor system that initiates apoptosis once cellular protection systems cannot cope anymore with cellular stress. PMID- 18413608 TI - Scent evolution in Chinese roses. AB - The phenolic methyl ether 3,5-dimethoxytoluene (DMT) is a major scent compound of many modern rose varieties, and its fragrance participates in the characteristic "tea scent" that gave their name to Tea and Hybrid Tea roses. Among wild roses, phenolic methyl ether (PME) biosynthesis is restricted to Chinese rose species, but the progenitors of modern roses included both European and Chinese species (e.g., Rosa chinensis cv Old Blush), so this trait was transmitted to their hybrid progeny. The last steps of the biosynthetic pathways leading to DMT involve two methylation reactions catalyzed by the highly similar orcinol O methyltransferases (OOMT) 1 and 2. OOMT1 and OOMT2 enzymes exhibit different substrate specificities that are consistent with their operating sequentially in DMT biosynthesis. Here, we show that these different substrate specificities are mostly due to a single amino acid polymorphism in the phenolic substrate binding site of OOMTs. An analysis of the OOMT gene family in 18 species representing the diversity of the genus Rosa indicated that only Chinese roses possess both the OOMT2 and the OOMT1 genes. In addition, we provide evidence that the Chinese-rose specific OOMT1 genes most probably evolved from an OOMT2-like gene that has homologues in the genomes of all extant roses. We propose that the emergence of the OOMT1 gene may have been a critical step in the evolution of scent production in Chinese roses. PMID- 18413609 TI - Distinct error-correcting and incidental learning of location relative to landmarks and boundaries. AB - Associative reinforcement provides a powerful explanation of learned behavior. However, an unproven but long-held conjecture holds that spatial learning can occur incidentally rather than by reinforcement. Using a carefully controlled virtual-reality object-location memory task, we formally demonstrate that locations are concurrently learned relative to both local landmarks and local boundaries but that landmark-learning obeys associative reinforcement (showing "overshadowing" and "blocking" or "learned irrelevance"), whereas boundary learning is incidental, showing neither overshadowing nor blocking nor learned irrelevance. Crucially, both types of learning occur at similar rates and do not reflect differences in levels of performance, cue salience, or instructions. These distinct types of learning likely reflect the distinct neural systems implicated in processing of landmarks and boundaries: the striatum and hippocampus, respectively [Doeller CF, King JA, Burgess N (2008) Proc Natl Acad Sci USA 105:5915-5920]. In turn, our results suggest the use of fundamentally different learning rules by these two systems, potentially explaining their differential roles in procedural and declarative memory more generally. Our results suggest a privileged role for surface geometry in determining spatial context and support the idea of a "geometric module," albeit for location rather than orientation. Finally, the demonstration that reinforcement learning applies selectively to formally equivalent aspects of task-performance supports broader consideration of two-system models in analyses of learning and decision making. PMID- 18413610 TI - Global regulatory logic for specification of an embryonic cell lineage. AB - Explanation of a process of development must ultimately be couched in the terms of the genomic regulatory code. Specification of an embryonic cell lineage is driven by a network of interactions among genes encoding transcription factors. Here, we present the gene regulatory network (GRN) that directs the specification of the skeletogenic micromere lineage of the sea urchin embryo. The GRN now includes all regulatory genes expressed in this lineage up to late blastula stage, as identified in a genomewide survey. The architecture of the GRN was established by a large-scale perturbation analysis in which the expression of each gene in the GRN was cut off by use of morpholinos, and the effects on all other genes were measured quantitatively. Several cis-regulatory analyses provided additional evidence. The explanatory power of the GRN suffices to provide a causal explanation for all observable developmental functions of the micromere lineage during the specification period. These functions are: (i) initial acquisition of identity through transcriptional interpretation of localized maternal cues; (ii) activation of specific regulatory genes by use of a double negative gate; (iii) dynamic stabilization of the regulatory state by activation of a feedback subcircuit; (iv) exclusion of alternative regulatory states; (v) presentation of a signal required by the micromeres themselves and of two different signals required for development of adjacent endomesodermal lineages; and (vi) lineage-specific activation of batteries of skeletogenic genes. The GRN precisely predicts gene expression responses and provides a coherent explanation of the biology of specification. PMID- 18413611 TI - Dephosphorylation of 2,3-bisphosphoglycerate by MIPP expands the regulatory capacity of the Rapoport-Luebering glycolytic shunt. AB - The Rapoport-Luebering glycolytic bypass comprises evolutionarily conserved reactions that generate and dephosphorylate 2,3-bisphosphoglycerate (2,3-BPG). For >30 years, these reactions have been considered the responsibility of a single enzyme, the 2,3-BPG synthase/2-phosphatase (BPGM). Here, we show that Dictyostelium, birds, and mammals contain an additional 2,3-BPG phosphatase that, unlike BPGM, removes the 3-phosphate. This discovery reveals that the glycolytic pathway can bypass the formation of 3-phosphoglycerate, which is a precursor for serine biosynthesis and an activator of AMP-activated protein kinase. Our 2,3-BPG phosphatase activity is encoded by the previously identified gene for multiple inositol polyphosphate phosphatase (MIPP1), which we now show to have dual substrate specificity. By genetically manipulating Mipp1 expression in Dictyostelium, we demonstrated that this enzyme provides physiologically relevant regulation of cellular 2,3-BPG content. Mammalian erythrocytes possess the highest content of 2,3-BPG, which controls oxygen binding to hemoglobin. We determined that total MIPP1 activity in erythrocytes at 37 degrees C is 0.6 mmol 2,3-BPG hydrolyzed per liter of cells per h, matching previously published estimates of the phosphatase activity of BPGM. MIPP1 is active at 4 degrees C, revealing a clinically significant contribution to 2,3-BPG loss during the storage of erythrocytes for transfusion. Hydrolysis of 2,3-BPG by human MIPP1 is sensitive to physiologic alkalosis; activity decreases 50% when pH rises from 7.0 to 7.4. This phenomenon provides a homeostatic mechanism for elevating 2,3-BPG levels, thereby enhancing oxygen release to tissues. Our data indicate greater biological significance of the Rapoport-Luebering shunt than previously considered. PMID- 18413612 TI - Alteration of cyclin D1 transcript elongation by a mutated transcription factor up-regulates the oncogenic D1b splice isoform in cancer. AB - Pre-mRNA splicing and polyadenylation are tightly connected to transcription, and transcriptional stimuli and elongation dynamics can affect mRNA maturation. However, whether this regulatory mechanism has a physio/pathological impact is not known. In cancer, where splice variant expression is often deregulated, many mutated oncogenes are transcriptional regulators. In particular, the Ewing sarcoma (EwSa) oncogene, resulting from a fusion of the EWS and FLI1 genes, encodes a well characterized transcription factor. EWS-FLI1 directly stimulates transcription of the CCND1 protooncogene encoding cyclin D1a and a less abundant but more oncogenic splice isoform, D1b. We show that, although both EWS and EWS FLI1 enhance cyclin D1 gene expression, they regulate the D1b/D1a transcript ratio in an opposite manner. Detailed analyses of RNA polymerase dynamics along the gene and of the effects of an inhibitor of elongation show that EWS-FLI1 favors D1b isoform expression by decreasing the elongation rate, whereas EWS has opposite effects. As a result, the D1b/D1a ratio is elevated in EwSa cell lines and tumors. The endogenous D1b protein is enriched in nuclei, where the oncogenic activity of cyclin D1 is known to occur, and depleting D1b in addition to D1a results in a stronger reduction of EwSa cell growth than depleting D1a only. These data show that elevated expression of a splice isoform in cancer can be due to an alteration of the transcription process by a mutated transcriptional regulator and provide evidence for a physio/pathological impact of the coupling between transcription and mRNA maturation. PMID- 18413614 TI - Cost-effective global conservation spending is robust to taxonomic group. AB - Priorities for conservation investment at a global scale that are based on a single taxon have been criticized because geographic richness patterns vary taxonomically. However, these concerns focused only on biodiversity patterns and did not consider the importance of socioeconomic factors, which must also be included if conservation funding is to be allocated efficiently. In this article, we create efficient global funding schedules that use information about conservation costs, predicted habitat loss rates, and the endemicity of seven different taxonomic groups. We discover that these funding allocation schedules are less sensitive to variation in taxon assessed than to variation in cost and threat. Two-thirds of funding is allocated to the same regions regardless of the taxon, compared with only one-fifth if threat and cost are not included in allocation decisions. Hence, if socioeconomic factors are considered, we can be more confident about global-scale decisions guided by single taxonomic groups. PMID- 18413613 TI - The evolutionarily conserved G protein-coupled receptor SREB2/GPR85 influences brain size, behavior, and vulnerability to schizophrenia. AB - The G protein-coupled receptor (GPCR) family is highly diversified and involved in many forms of information processing. SREB2 (GPR85) is the most conserved GPCR throughout vertebrate evolution and is expressed abundantly in brain structures exhibiting high levels of plasticity, e.g., the hippocampal dentate gyrus. Here, we show that SREB2 is involved in determining brain size, modulating diverse behaviors, and potentially in vulnerability to schizophrenia. Mild overexpression of SREB2 caused significant brain weight reduction and ventricular enlargement in transgenic (Tg) mice as well as behavioral abnormalities mirroring psychiatric disorders, e.g., decreased social interaction, abnormal sensorimotor gating, and impaired memory. SREB2 KO mice showed a reciprocal phenotype, a significant increase in brain weight accompanying a trend toward enhanced memory without apparent other behavioral abnormalities. In both Tg and KO mice, no gross malformation of brain structures was observed. Because of phenotypic overlap between SREB2 Tg mice and schizophrenia, we sought a possible link between the two. Minor alleles of two SREB2 SNPs, located in intron 2 and in the 3' UTR, were overtransmitted to schizophrenia patients in a family-based sample and showed an allele load association with reduced hippocampal gray matter volume in patients. Our data implicate SREB2 as a potential risk factor for psychiatric disorders and its pathway as a target for psychiatric therapy. PMID- 18413615 TI - The influence of cochlear shape on low-frequency hearing. AB - The conventional theory about the snail shell shape of the mammalian cochlea is that it evolved essentially and perhaps solely to conserve space inside the skull. Recently, a theory proposed that the spiral's graded curvature enhances the cochlea's mechanical response to low frequencies. This article provides a multispecies analysis of cochlear shape to test this theory and demonstrates that the ratio of the radii of curvature from the outermost and innermost turns of the cochlear spiral is a significant cochlear feature that correlates strongly with low-frequency hearing limits. The ratio, which is a measure of curvature gradient, is a reflection of the ability of cochlear curvature to focus acoustic energy at the outer wall of the cochlear canal as the wave propagates toward the apex of the cochlea. PMID- 18413616 TI - Conservation of the regulated structure of folded myosin 2 in species separated by at least 600 million years of independent evolution. AB - The myosin 2 family of molecular motors includes isoforms regulated in different ways. Vertebrate smooth-muscle myosin is activated by phosphorylation of the regulatory light chain, whereas scallop striated adductor-muscle myosin is activated by direct calcium binding to its essential light chain. The paired heads of inhibited molecules from myosins regulated by phosphorylation have an asymmetric arrangement with motor-motor interactions. It was unknown whether such interactions were a common motif for inactivation used in other forms of myosin linked regulation. Using electron microscopy and single-particle image processing, we show that indistinguishable structures are indeed found in myosins and heavy meromyosins isolated from scallop striated adductor muscle and turkey gizzard smooth muscle. The similarities extend beyond the shapes of the heads and interactions between them: In both myosins, the tail folds into three segments, apparently at identical sites; all three segments are in close association outside the head region; and two segments are associated in the same way with one head in the asymmetric arrangement. Thus, these organisms, which have different regulatory mechanisms and diverged from a common ancestor >600 Myr ago, have the same quaternary structure. Conservation across such a large evolutionary distance suggests that this conformation is of fundamental functional importance. PMID- 18413617 TI - Endogenous steroids and financial risk taking on a London trading floor. AB - Little is known about the role of the endocrine system in financial risk taking. Here, we report the findings of a study in which we sampled, under real working conditions, endogenous steroids from a group of male traders in the City of London. We found that a trader's morning testosterone level predicts his day's profitability. We also found that a trader's cortisol rises with both the variance of his trading results and the volatility of the market. Our results suggest that higher testosterone may contribute to economic return, whereas cortisol is increased by risk. Our results point to a further possibility: testosterone and cortisol are known to have cognitive and behavioral effects, so if the acutely elevated steroids we observed were to persist or increase as volatility rises, they may shift risk preferences and even affect a trader's ability to engage in rational choice. PMID- 18413618 TI - Safety and efficacy of a testosterone metered-dose transdermal spray for treating decreased sexual satisfaction in premenopausal women: a randomized trial. AB - BACKGROUND: It is not known whether premenopausal women who report low sexual satisfaction and have low circulating testosterone levels will benefit from testosterone therapy. OBJECTIVE: To evaluate the effects of exogenous testosterone in premenopausal women reporting diminished sexual function. DESIGN: Randomized, double-blind, placebo-controlled, dose-ranging trial. SETTING: 6 Australian medical centers. PATIENTS: 261 women age 35 to 46 years who reported a decrease in satisfying sexual activity relative to their younger years and had a morning serum free testosterone level less than 3.8 pmol/L (<1.1 pg/mL). INTERVENTION: 3 different doses of testosterone administered by a metered-dose transdermal spray for 16 weeks or placebo. MEASUREMENTS: The primary outcome was the mean number of self-reported satisfactory sexual events (SSEs) over 28 days at week 16. The frequency of SSEs, total number of sexual events (every 4 weeks), scores from the modified Sabbatsberg Sexual Self-Rating Scale and the Psychological General Well-Being Index, and safety variables were also measured. RESULTS: The number of SSEs increased during the treatment period in the active treatment groups and the placebo group. The mean number of SSEs over 28 days at week 16 was statistically significantly greater for women treated with the intermediate dose of testosterone therapy (one 90-microL spray) than for women treated with placebo. The least-squares mean was 2.48 versus 1.70 SSEs, respectively (event rate ratio, 1.49 [95% CI, 1.01 to 2.18]; P = 0.04). The frequency of SSEs in women treated with low and high doses of testosterone did not differ from that in women who took placebo. The rate ratios based on the least-squares mean rates of SSEs during weeks 4 to 16 for each treatment group showed statistically significant or borderline significant increases in all testosterone groups compared with the placebo group. The rate ratios for the one 56-microL spray, one 90-microL spray, and two 90-microL sprays treatment groups were 1.34 (CI, 0.97 to 1.85; P = 0.081), 1.48 (CI, 1.07 to 2.06; P = 0.018), and 1.38 (CI, 1.00 to 1.92; P = 0.052), respectively. At week 16, 95% of women treated with the one 90-microL dose had a free testosterone level less than the upper limit of the reference range for women. The most frequently reported adverse event was hypertrichosis, which was dose-related and mostly confined to the application site. No clinically relevant changes in blood test values, serum biochemical variables, or vital signs occurred. LIMITATION: The study duration was short, and the placebo effect was strong. CONCLUSION: A daily 90-microL dose of transdermal testosterone improves self-reported sexual satisfaction for premenopausal women with reduced libido and low serum-free testosterone levels by a mean of 0.8 SSE per month. The rate of SSEs with higher and lower testosterone doses did not differ from that with placebo. PMID- 18413619 TI - Effect of unrelated comorbid conditions on hypertension management. AB - BACKGROUND: Quality-of-care assessment at a single visit can be affected by whether a patient's comorbid conditions are related or unrelated to a specific measure. OBJECTIVE: To examine the association of unrelated comorbid conditions with treatment of uncontrolled hypertension in primary care visits. DESIGN: Examination of a database derived from electronic medical records collected during routine care of a cohort of primary care patients. SETTING: 6 primary care practices in Philadelphia, Pennsylvania. PATIENTS: 15,459 patients with uncontrolled hypertension who made 70,557 visits to 200 clinicians from January 2004 through December 2006. MEASUREMENTS: Intensification of any antihypertensive treatment before the next visit was assessed. Patient and clinician information were obtained from electronic medical records and administrative data. Unrelated comorbid conditions included 28 conditions, such as arthritis and emphysema, whereas related comorbid conditions included vascular diseases. Generalized estimating equation logistic regression models were used to adjust for patient, health care, and provider characteristics and for clustering. Variation in the effect of unrelated comorbid conditions was examined at the visit, patient, and provider level. RESULTS: At study visits, patients had a mean of 2.2 (SD, 1.8) unrelated comorbid conditions. The adjusted odds of treatment intensification decreased with the number of unrelated comorbid conditions, from 0.85 (95% CI, 0.80 to 0.90) for 1 to 0.59 (CI, 0.51 to 0.69) for 7 or more versus none. The relationship between treatment intensification and unrelated comorbid conditions persisted at the visit, patient, and provider levels (P < 0.001). LIMITATIONS: The reasons for not intensifying treatments are unknown. The recorded blood pressure may be inaccurate. Physicians may vary in their recording of comorbid conditions. CONCLUSION: Patients with more unrelated comorbid conditions were less likely to have uncontrolled hypertension addressed at a visit. The effect of different types of comorbid conditions on meeting quality-of-care measures merits further investigation. PMID- 18413620 TI - Narrative review: paroxysmal nocturnal hemoglobinuria: the physiology of complement-related hemolytic anemia. PMID- 18413621 TI - Meta-analysis: antibiotics for prophylaxis against hemodialysis catheter-related infections. AB - BACKGROUND: Catheter-related infections cause morbidity and mortality in patients undergoing hemodialysis. PURPOSE: To examine whether topical or intraluminal antibiotics reduce catheter-related bloodstream infection compared with no antibiotic therapy in adults undergoing hemodialysis. DATA SOURCES: Electronic databases, trial registries, bibliographies, and conference proceedings up to October 2007, with no language restrictions. STUDY SELECTION: Two reviewers independently selected randomized, controlled trials using topical or intraluminal antibiotics for prophylaxis of infection in adults with catheters who are undergoing hemodialysis. DATA EXTRACTION: Two independent reviewers assessed studies for inclusion, quality, and extracted data. DATA SYNTHESIS: Fixed-effects models were used to estimate pooled rate ratios for outcomes. Topical antibiotics reduced the rate of bacteremia (rate ratio, 0.22 [95% CI, 0.12 to 0.40]; 0.10 vs. 0.45 case of bacteremia per 100 catheter-days), exit-site infection (rate ratio, 0.17 [CI, 0.08 to 0.38]; 0.06 vs. 0.41 case of infection per 100 catheter-days), need for catheter removal, and hospitalization for infection. Intraluminal antibiotics reduced the rate of bacteremia (rate ratio, 0.32 [CI, 0.22 to 0.47]; 0.12 vs. 0.32 case of bacteremia per 100 catheter-days) and need for catheter removal. Intraluminal antibiotics did not significantly reduce the rate of exit-site infection, and no hospitalization data were available for these agents. LIMITATIONS: The evidence base included only 16 trials, and most had less than 6 months of follow-up. Only one third of studies were blinded. Publication bias was evident. CONCLUSION: Both topical and intraluminal antibiotics reduced the rate of bacteremia as well as the need for catheter removal secondary to complications. Whether these strategies will lead to antimicrobial resistance and loss of efficacy over longer periods remains unclear. PMID- 18413622 TI - Emerging antimicrobial resistance in Neisseria gonorrhoeae: urgent need to strengthen prevention strategies. AB - Prevention and control of gonorrhea is an important public health concern due to the high burden of disease, the recent increase in reported infection rates, and the reproductive and economic consequences of infection. Effective antibiotic treatment is one essential component of an integrated approach to gonorrhea control. Over the past 60 years, however, development of resistance in Neisseria gonorrhoeae to multiple antimicrobial classes challenges this component of gonorrhea control. An integrated, comprehensive prevention strategy should include enhancement of national and international surveillance systems to monitor antimicrobial resistance and new strategies to maximize the benefit and prolong the utility of antimicrobials, including combination regimens, implementation of screening recommendations for individuals at high risk for infection, and the assurance of prompt and effective treatment for infected persons and their sexual partners. Progress in controlling the epidemic and avoiding a resurgence as treatment options wane will require careful attention to all components of a comprehensive prevention strategy. PMID- 18413623 TI - The international pharmaceutical market as a source of low-cost prescription drugs for U.S. patients. AB - In response to increasing prescription drug costs, more U.S. patients and policymakers are importing less-expensive pharmaceutical products from other countries. Large-scale prescription drug importation is currently illegal, but the U.S. Food and Drug Administration permits individuals to bring in 90-day supplies of drugs for personal use. As patient use of foreign-bought drugs has increased, federal legislators have continued to debate the full legalization of importation. Three factors help guide whether U.S. patients and policymakers can rely on other countries as sources of imported prescription drugs: whether the safety of the product can be ensured, how the import price compares with domestic prices, and how importation might affect the exporting country's pharmaceutical market. In wealthier countries with active regulatory systems, drug safety can be adequately ensured, and brand-name products are usually less expensive than in the United States (although generic drugs may be more expensive). However, implementing large-scale importation can negatively impact the originating country's market and can diminish the long-term cost savings for U.S. consumers. In low- and middle-income countries, prices may be reduced for both brand-name and generic drugs, but the prevalence of unauthorized products on the market makes ensuring drug safety more difficult. It may be reasonable for individual U.S. consumers to purchase essential medicines from certain international markets, but the most effective way to decrease drug costs overall is the appropriate use of domestic generic drugs, which are available for almost every major therapeutic class. PMID- 18413624 TI - Testosterone supplementation to improve women's sexual satisfaction: complexities and unknowns. PMID- 18413625 TI - Paying at the pump. PMID- 18413626 TI - Could increasing the duration of triple therapy be a clinically useful strategy? PMID- 18413627 TI - When to switch therapy in patients with severe community-acquired pneumonia. PMID- 18413628 TI - Does tiotropium reduce hospitalizations in chronic obstructive pulmonary disease? PMID- 18413629 TI - How to improve coordination of care. PMID- 18413630 TI - Is it safe to conclude that Beers criteria medications led to few adverse events? PMID- 18413631 TI - How to improve coordination of care. PMID- 18413632 TI - Are pay-for-performance programs a threat to medical professionalism? PMID- 18413633 TI - Are pay-for-performance programs a threat to medical professionalism? PMID- 18413634 TI - Are pay-for-performance programs a threat to medical professionalism? PMID- 18413635 TI - Memantine-induced hepatitis with cholestasis in a very elderly patient. PMID- 18413636 TI - Successful treatment of fulminant Clostridium difficile infection with fecal bacteriotherapy. PMID- 18413637 TI - Overanticoagulation with coumarin and cutaneous azole therapy. PMID- 18413638 TI - Ten-year follow-up of radiation therapy oncology group protocol 92-02: a phase III trial of the duration of elective androgen deprivation in locally advanced prostate cancer. AB - PURPOSE: To determine whether adding 2 years of androgen-deprivation therapy (ADT) improved outcome for patients electively treated with ADT before and during radiation therapy (RT). PATIENTS AND METHODS: Prostate cancer patients with T2c T4 prostate cancer with no extra pelvic lymph node involvement and prostate specific antigen (PSA) less than 150 ng/mL were included. All patients received 4 months of goserelin and flutamide before and during RT. They were randomized to no further ADT (short-term ADT [STAD] + RT) or 24 months of goserelin (long-term ADT [LTAD] + RT). A total of 1,554 patients were entered. RT was 45 Gy to the pelvic nodes and 65 to 70 Gy to the prostate. Median follow-up of all survival patients is 11.31 and 11.27 years for the two arms. RESULTS: At 10 years, the LTAD + RT group showed significant improvement over the STAD + RT group for all end points except overall survival: disease-free survival (13.2% v 22.5%; P < .0001), disease-specific survival (83.9% v 88.7%; P = .0042), local progression (22.2% v 12.3%; P < .0001), distant metastasis (22.8% v 14.8%; P < .0001), biochemical failure (68.1% v 51.9%; P 60 years, WHO performance status of 2, or elevated serum lactate dehydrogenase). Four doses of rituximab were infused on days -7, 1, 22, and 43, and CHOP was administered on days 3, 24, and 45, followed 3 weeks later by 40 to 46 Gy of IFRT. RESULTS: Sixty patients with aggressive NHL were eligible. With the median follow-up of 5.3 years, treatment resulted in a progression-free survival (PFS) of 93% at 2 years and 88% at 4 years. Overall survival (OS) was 95% at 2 years and 92% at 4 years. These results were compared with those from a historic group of patients treated without rituximab on S8736, demonstrating PFS of 78% and OS of 88% at 4 years. CONCLUSION: In limited-stage DLBCL, the addition of rituximab to three cycles of CHOP plus IFRT met prespecified study criteria of efficacy, with 2-year PFS of at least 84%, meriting further investigation. There is a pattern of continuing relapse with modest survival gains. We hypothesize that such a pattern may be the result of biologic differences between limited- and advanced-stage lymphoma. PMID- 18413641 TI - Intensive chemotherapy followed by hematopoietic stem-cell rescue for refractory and recurrent primary CNS and intraocular lymphoma: Societe Francaise de Greffe de Moelle Osseuse-Therapie Cellulaire. AB - PURPOSE: The prognosis of relapsing primary CNS lymphoma (PCNSL) is poor. We report the results of a prospective multicenter trial of intensive chemotherapy followed by autologous hematopoietic stem-cell rescue (IC + HCR) in immunocompetent adult patients with PCNSL or intraocular lymphoma (IOL) after failure of high-dose methotrexate-based treatment. PATIENTS AND METHODS: Salvage treatment consisted of two cycles of high-dose cytarabine and etoposide (CYVE). Intensive chemotherapy combined thiotepa, busulfan, and cyclophosphamide. Forty three patients (median age, 52 years; range, 23 to 65 years) were included, with relapse (n = 22), refractory disease (n = 17), or a partial response to first line treatment (n = 4). The response to CYVE was not assessable in three cases because of treatment-related death. Twenty patients (47%) were chemosensitive to CYVE: 15 of them proceeded to IC + HCR. IC + HCR was also administered to 12 patients who did not respond to CYVE. All but one of the 27 patients who underwent IC + HCR entered complete remission. RESULTS: With a median follow-up of 36 months, the median overall survival was 18.3 months in the overall population, and 58.6 months among patients who completed IC + HCR. The respective median progression-free survival (PFS) times after IC + HCR were 11.6 and 41.1 months. The 2-year overall survival probability was 45% in the whole population and 69% among the 27 patients who received IC + HCR. The 2-year PFS probability was 43% among all the patients and 58% in the IC + HCR subpopulation. CONCLUSION: IC + HCR is an effective treatment for refractory and recurrent PCNSL. PMID- 18413642 TI - Factors affecting response and survival in patients with myelodysplasia treated with immunosuppressive therapy. AB - PURPOSE: Marrow failure in some patients with myelodysplastic syndrome (MDS) responds to immunosuppressive treatment (IST), but long-term outcome after IST has not been described. We evaluated patients with MDS treated with IST at our institution to determine their clinical course compared with a comparable supportive care only group. PATIENTS AND METHODS: One hundred twenty-nine patients with MDS received IST with a median follow-up of 3.0 years (range, 0.03 to 11.3 years), using antithymocyte globulin (ATG) or cyclosporine (CsA) in combination or singly. Variables affecting response and survival were studied and outcomes were compared with those of 816 patients with MDS reported to the International Myelodysplasia Risk Analysis Workshop (IMRAW) who received only supportive care. RESULTS: Thirty-nine (30%) of 129 patients receiving IST responded either completely or partially: 18 (24%) of 74 patients responded to ATG, 20 (48%) of 42 patients responded to ATG plus CsA, and one (8%) of 13 patients responded to CsA. Thirty-one percent (12 of 39) of the responses were complete, resulting in transfusion independence and near-normal blood counts. In multivariate analysis, younger age was the most significant factor favoring response to therapy. Other favorable factors affecting response were HLA-DR15 positivity and combination ATG plus CsA treatment (P = .001 and P = .048, respectively). In multivariate analysis of the combined IMRAW and IST cohorts, younger age, treatment with IST, and intermediate or low International Prognostic Scoring System score significantly favored survival. CONCLUSION: IST produced significant improvement in the pancytopenia of a substantial proportion of patients with MDS and was associated with improved overall and progression-free survival, especially in younger individuals with lower-risk disease. PMID- 18413643 TI - Abstracts of Posters Presented at the 18th Annual Meeting & 28th Computers in Anesthesia Meeting of the Society for Technology in Anesthesia. San Diego, California, USA. January 16-19, 2008. PMID- 18413645 TI - Keratinocyte migration, proliferation, and differentiation in chronic ulcers from patients with diabetes and normal wounds. AB - Epithelialization of normal acute wounds occurs by an orderly series of events whereby keratinocytes migrate, proliferate, and differentiate to restore barrier function. The keratinocytes in the epidermis of chronic ulcers fail to execute this series of events. To better understand the epithelial dynamics of chronic ulcers, we used immunohistochemistry to evaluate proliferation, differentiation, adhesion, and migration in keratinocytes along the margin of chronic ulcers from patients with diabetes mellitus. We compared these features with keratinocytes from the migrating epithelial tongues of acute incisional and excisional wounds from normal volunteers. Keratinocytes at the chronic ulcer edge are highly proliferative (Ki67 proliferation marker), have an activated phenotype (K16), do not stain for keratins involved in epidermal differentiation (K10 and K2), and show a reduced expression of LM-3A32 (uncleaved, precursor of the alpha3 chain of laminin 5), a key molecule present on migrating epithelium. In contrast, keratinocytes in normal acute wound migrating epithelium do not express the proliferation marker Ki67 but do express K10, K2, and LM-3A32. A better understanding of molecular mechanisms involved in keratinocyte migration may lead to molecular targets for therapies for impaired wound healing. PMID- 18413646 TI - Phenotypic and genetic characterization of circulating tumor cells by combining immunomagnetic selection and FICTION techniques. AB - The presence of circulating tumor cells (CTCs) in breast cancer patients has been proven to have clinical relevance. Cytogenetic characterization of these cells could have crucial relevance for targeted cancer therapies. We developed a method that combines an immunomagnetic selection of CTCs from peripheral blood with the fluorescence immunophenotyping and interphase cytogenetics as a tool for investigation of neoplasm (FICTION) technique. Briefly, peripheral blood (10 ml) from healthy donors was spiked with a predetermined number of human breast cancer cells. Nucleated cells were separated by double density gradient centrifugation of blood samples. Tumor cells (TCs) were immunomagnetically isolated with an anti cytokeratin antibody and placed onto slides for FICTION analysis. For immunophenotyping and genetic characterization of TCs, a mixture of primary monoclonal anti-pancytokeratin antibodies was used, followed by fluorescent secondary antibodies, and finally hybridized with a TOP2A/HER-2/CEP17 multicolor probe. Our results show that TCs can be efficiently isolated from peripheral blood and characterized by FICTION. Because genetic amplification of TOP2A and ErbB2 (HER-2) in breast cancer correlates with response to anthracyclines and herceptin therapies, respectively, this novel methodology could be useful for a better classification of patients according to the genetic alterations of CTCs and for the application of targeted therapies. PMID- 18413647 TI - Production and histological application of affinity-purified antibodies to heat denatured green fluorescent protein. AB - Enhanced green fluorescent protein (GFP) irreversibly loses not only fluorescence but also antigenicity recognized with conventional anti-GFP antibodies by heat denaturation. This hinders combinatory applications of the GFP immunodetection technique with heat-requiring procedures, such as in situ hybridization histochemistry, antigen retrieval, and Western blot. Here we produced new rabbit and guinea pig antibodies against heat-denatured GFP. The polyclonal antibodies affinity-purified with the antigen column detected a single band corresponding to the molecular size of GFP in Western blot analysis, with mouse brain expressing GFP from the GAD67 locus. By immunofluorescence labeling, the new antibodies detected GFP molecules in heat (> or = 70 degrees C)-treated sections but not in untreated sections of the mouse brain. When the sections were incubated at > or = 37 degrees C with in situ hybridization buffer containing 50% formamide, a denaturing reagent, the sections lost immunoreactivity with the conventional anti GFP antibodies but acquired immunoreactivity with the new antibodies to heat denatured GFP. Finally, GFP immunofluorescence was successfully visualized with the new antibodies in sections of the GFP-expressing mice labeled by fluorescence in situ hybridization histochemistry against GAD67 mRNA. Thus, the antibodies produced in this study may provide an opportunity to combine GFP immunodetection with procedures requiring heat treatment. This manuscript contains online supplemental material at http://www.jhc.org. Please visit this article online to view these materials. PMID- 18413649 TI - C9orf10 protein, a novel protein component of Puralpha-containing mRNA-protein particles (Puralpha-mRNPs): characterization of developmental and regional expressions in the mouse brain. AB - Puralpha has been implicated in mRNA transport and translation in neurons. We previously reported that Puralpha is a component of mRNA/protein complexes (Puralpha-mRNPs) with several other proteins. Among them, we found the C9orf10 (Homo sapiens chromosome 9 open reading frame 10) protein, which was recently characterized as a component of RNA-containing structures. However, C9orf10 itself remains poorly understood. To characterize C9orf10 expression at the protein level, we raised an antibody against C9orf10 and compared the spatial and developmental expressions of this protein and Puralpha in the mouse brain. C9orf10 was expressed as early as embryo stage 12, whereas Puralpha was expressed from 5 days after birth. In adults, C9orf10 expression was most prominent in the hippocampus, caudate putamen, cerebral cortex, and cerebellum, unlike the uniform distribution of Puralpha. C9orf10-positive cells also showed immunoreactivity to Puralpha. C9orf10 expression was restricted to neurons, judging by the immunoreactivity to neuron-specific nuclear protein or CaM kinase II. These observations suggest an accessory role of C9orf10 for Puralpha in a limited brain region in addition to other possible functions that have not yet been determined. PMID- 18413648 TI - Leptin and the obesity receptor (OB-R) in the small intestine and colon: a colocalization study. AB - Leptin is a hormone that plays an important role in overall body energy homeostasis, and the obesity receptor, OB-R, is widely distributed in the organism. In the intestine, a multitude of leptin actions have been reported, but it is currently unclear to what extent the hormone affects the intestinal epithelial cells by an endocrine or exocrine signaling pathway. To elucidate this, the localization of endogenous porcine leptin and OB-R in enterocytes and colonocytes was studied. By immunofluorescence microscopy, both leptin and OB-R were mainly observed in the basolateral membrane of enterocytes and colonocytes but also in the apical microvillar membrane of the cells. By electron microscopy, coclustering of hormone and receptor in the plasma membrane and localization in endosomes was frequently detected at the basolateral surface of the epithelial cells, indicative of leptin signaling activity. In contrast, coclustering occurred less frequently at the apical cell surface, and subapical endosomal localization was hardly detectable. We conclude that leptin action in intestinal epithelial cells takes place at the basolateral plasma membrane, indicating that the hormone uses an endocrine pathway both in the jejunum and colon. In contrast, the data obtained did not provide evidence for an exocrine, lumenal action of the hormone in the intestine. PMID- 18413650 TI - Differential expression of decorin by human malignant and benign vascular tumors. AB - An increasing amount of evidence indicates that a small extracellular chondroitin/dermatan sulfate proteoglycan, decorin, is indirectly involved in angiogenesis. Given that angiogenesis is a sine qua non for tumor growth and progression, we attempted to examine whether human malignant vascular tumors differ from human benign vascular tumors in terms of their decorin expression and synthesis. CD31 immunostaining demonstrated that the human malignant vascular tumors Kaposi's sarcoma and angiosarcoma were filled with capillary-like structures, whereas in benign cavernous and capillary hemangiomas, blood vessels were not as abundantly present. By utilizing in situ hybridization and immunocytochemical assays for decorin, we showed that there was no detectable decorin mRNA expression or immunoreactivity within the tumor mass in the Kaposi's sarcoma or angiosarcoma group. Instead, decorin was expressed in the connective tissue stroma lining the sarcoma tissue. In contrast to sarcomas, in hemangiomas, decorin mRNA expression and immunoreactivity were observed also within the tumor mass, particularly in the connective tissue stroma surrounding the clusters of intratumoral blood vessels. Finally, distribution of type I collagen was found to be similar to that of decorin in these tumor tissues. Our findings can be explained with different states of angiogenesis in dissimilar growths. In sarcomas, angiogenesis is extremely powerful, whereas in hemangiomas, angiogenesis has ceased. Thus, decorin is likely to possess a suppressive effect on human tumor angiogenesis in vivo, as previously described by studies using different experimental models. Decorin certainly provides a usable biomarker for distinguishing between benign and malignant vascular tumors in patients. PMID- 18413651 TI - Site-specific expression of IQGAP1, a key mediator of cytoskeleton, in mouse renal tubules. AB - IQGAP1 is a multifunctional junction molecule that is involved in cell migration, proliferation, differentiation, cell polarity, and cell-cell adhesion. It is highly expressed in the kidney and has recently been identified in the glomerular basement membrane as a nephrin-associated protein. However, the distribution of IQGAP1 in renal tubular epithelial cells is unknown. We performed confocal microscopic studies to localize IQGAP1 in each nephron segment using dual immunofluorescence staining with various antibodies against segment-specific markers. We found that IQGAP1 was strongly expressed in the distal convoluted tubule (DCT), collecting duct, and macula densa and moderately in the thick ascending limb and proximal tubule. In the DCT, the IQGAP1-F-actin complex forms a comb-like structure with multiple parallel spikes sitting on the basal membrane. In the macula densa cells, IQGAP1 is strongly expressed in the apical membrane, whereas in type A intercalated cells, IQGAP1 is expressed in the basolateral membrane, where it colocalizes with anion exchanger 1, and in principal cells, it is diffusely expressed. In conclusion, we showed the expression and subcellular localization of IQGAP1 in various nephron segments. The site-specific expression pattern of this potent modulator of multiple biological pathways in the renal tubules suggests that IQGAP1 may have multiple important roles in various renal functions. PMID- 18413652 TI - Rapid microwave fixation of cell monolayers preserves microtubule-associated cell structures. AB - Microwave (MW) fixation has been suggested as a method to rapidly immobilize cellular dynamics for fine structural studies in the electron microscope. To show its suitability for studies on cell monolayers, one has to apply MW fixation systematically in correlation with samples on the light microscopy level. Examples for MW fixation of cell monolayers, however, are still rare. MW accelerated fixation for relatively long periods of time (1-2 min) has been reported without showing its suitability at the fine structural level. Here, we provide a rapid MW fixation protocol for cell monolayers on a subminute time scale. The impact of the MW-accelerated glutaraldehyde fixation on temperature sensitive cytoskeletal components such as microtubules was evaluated. For testing the effectiveness of MW-assisted primary fixation, saponin treatment of the monolayers was included. Simultaneous MW-accelerated fixation and extraction by saponin was necessary to achieve a gradual improvement in visualization of cytoskeletal aspects in association with cell junctions, mitochondria, and centrioles. To establish a valuable routine program for fine structural studies of resin-embedded cell models on substrata, a protocol combining MW fixation with automatic processing in a tissue processor is provided. PMID- 18413653 TI - Evaluation of chromogenic MRSA medium, MRSA select and Oxacillin Resistance Screening Agar for the detection of methicillin-resistant Staphylococcus aureus. AB - AIMS: An evaluation of Chromogenic MRSA medium (CMRSA), MRSASelect (MRSAS) and Oxacillin Resistance Screening Agar (ORSA) was performed to determine the optimum medium providing a rapid and sensitive method for methicillin-resistant Staphylococcus aureus (MRSA) detection. METHODS: A total of 632 clinical specimens were cultured on the three media in first phase of the study, while 720 clinical specimens were cultured on CMRSA and ORSA in the second phase. RESULTS: The sensitivity and specificity, respectively, of the media in the first phase were: CMRSA 88.9% and 98.45%; MRSAS 92.1% and 99.1%; ORSA (24 h incubation) 68.3% and 98.8%; and ORSA (48 h incubation) 85.7% and 96.3%. In the second phase the sensitivity and specificity, respectively, were CMRSA 91.2% and 98.6%; ORSA (24 h incubation) 58.9% and 98.2%; and ORSA (48 h incubation) 85.6% and 95.6%. The positive predictive values of the two chromogenic media were higher than that for ORSA. There were fewer false-positive results with the chromogenic media (1.4% for CMRSA and 0.8% for MRSAS) compared with ORSA (3.3%). CONCLUSIONS: Performing latex agglutination tests on growth from chromogenic media provides results for 93.8% of MRSA isolates within 24 h. There is a small increase in cost of chromogenic media compared with ORSA ( pound28 for MRSAS, and pound36 for CMRSA, per 1000 specimens) and direct agglutination tests ( pound80 per 1000 specimens). However early availability of MRSA screening results can reduce the burden of MRSA in hospitals because of early implementation of infection control measures. PMID- 18413654 TI - FOXP3 immunohistochemistry on formalin-fixed paraffin-embedded tissue: poor correlation between different antibodies. AB - Since its original description, there has been a substantial output of publications related to the FOXP3 gene. The FOXP3 protein, a member of the forkhead/winged-helix family of transcriptional regulators is a nuclear product and is not expressed in the cell cytoplasm or on the cell surface. Expression of this single transcription factor causes a developmental switch in naive T cells to a suppressor cell phenotype, more commonly referred to as regulatory T cells (Tregs). Tregs have been intensively studied in various autoimmune diseases, infections and different cancers. An increasing choice of commercially available monoclonal antibodies targeting FOXP3 is now available. This report describes the experience of using two commonly used monoclonal FOXP3 antibodies on formalin fixed paraffin-embedded sections of different organs, including the cervix and vulva. The antibodies targeting different FOXP3 epitopes unexpectedly resulted in significantly different staining patterns. This phenomenon has not been previously reported and is likely to be an important observation. PMID- 18413655 TI - Relationships of the woody Medicago species (section Dendrotelis) assessed by molecular cytogenetic analyses. AB - BACKGROUND AND AIMS: The organization of rDNA genes in the woody medic species from the agronomically important Medicago section Dendrotelis was analysed to gain insight into their taxonomic relationships, to assess the levels of infraspecific variation concerning ribosomal loci in a restricted and fragmented insular species (M. citrina) and to assess the nature of its polyploidy. METHODS: Fluorescence in situ hybridization (FISH) was used for physical mapping of 5S and 45S ribosomal DNA genes in the three species of section Dendrotelis (M. arborea, M. citrina, M. strasseri) and the related M. marina from section Medicago. Genomic in situ hybridization (GISH) was used to assess the genomic relationships of the polyploid M. citrina with the putatively related species from section Dendrotelis. KEY RESULTS: The diploid (2n = 16) M. marina has a single 45S and two 5S rDNA loci, a pattern usually detected in previous studies of Medicago diploid species. However, polyploid species from section Dendrotelis depart from expectations. The tetraploid species (2n = 32) M. arborea and M. strasseri have one 45S rDNA locus and two 5S rDNA loci, whereas in the hexaploid (2n = 48) M. citrina four 45S rDNA and five 5S rDNA loci have been detected. No single chromosome of M. citrina was uniformly labelled after using genomic probes from M. arborea and M. strasseri. Instead, cross-hybridization signals in M. citrina were restricted to terminal chromosome arms and NOR regions. CONCLUSIONS: FISH results support the close taxonomic interrelationship between M. arborea and M. strasseri. In these tetraploid species, NOR loci have experienced a diploidization event through physical loss of sequences, a cytogenetic feature so far not reported in other species of the genus. The high number of rDNA loci and GISH results support the specific status for the hexaploid M. citrina, and it is suggested that this species is not an autopolyploid derivative of M. arborea or M. strasseri. Further, molecular cytogenetic data do not suggest the hypothesis that M. arborea and M. strasseri were involved in the origin of M. citrina. FISH mapping can be used as an efficient tool to determine the genomic contribution of M. citrina in somatic hybrids with other medic species. PMID- 18413656 TI - Anatomy, ultrastructure and chemical composition of food bodies of Hovenia dulcis (Rhamnaceae). AB - BACKGROUND AND AIMS: Food bodies (FBs) are structures that promote mutualism between plants and ants, which help protect them against herbivores. The present study aims to describe the anatomical organization, ultrastructure and chemical composition of the FBs in Hovenia dulcis, which represent the first structures of this type described in Rhamnaceae. METHODS: Leaves in various stages of development were collected and fixed for examination under light, transmission and scanning electron microscopy. Samples of FBs were subjected to chemical analysis using thin-layer chromatography and nuclear magnetic resonance of (1)H and (13)C. KEY RESULTS: The FBs vary from globose to conical and are restricted to the abaxial leaf surface, having a mixed origin, including epidermis and parenchyma. The FB epidermis is uniseriate, slightly pilose and has a thin cuticle. The epidermal cells are vacuolated and pigments or food reserves are absent. The parenchyma cells of immature FBs have dense cytoplasm showing mitochondria, endoplasmic reticulum and plastids. Mature FB cells store oils, which are free in the cytosol and occupy a large portion of the cell lumen. In these cells the plastids accumulate starch. CONCLUSIONS: The lipids present in FBs are glycerin esters characteristic of plant energy reserves. Ants were observed collecting these FBs, which allows us to infer that these structures mediate plant-ant interactions and can help protect the young plants against herbivores, as these structures are prevalent at this developmental stage. PMID- 18413657 TI - Evidence-based guidelines for treating depressive disorders with antidepressants: a revision of the 2000 British Association for Psychopharmacology guidelines. AB - A revision of the 2000 British Association for Psychopharmacology evidence-based guidelines for treating depressive disorders with antidepressants was undertaken to incorporate new evidence and to update the recommendations where appropriate. A consensus meeting involving experts in depressive disorders and their management was held in May 2006. Key areas in treating depression were reviewed, and the strength of evidence and clinical implications were considered. The guidelines were drawn up after extensive feedback from participants and interested parties. A literature review is provided, which identifies the quality of evidence to inform the recommendations, the strength of which are based on the level of evidence. These guidelines cover the nature and detection of depressive disorders, acute treatment with antidepressant drugs, choice of drug versus alternative treatment, practical issues in prescribing and management, next-step treatment, relapse prevention, treatment of relapse, and stopping treatment. PMID- 18413658 TI - Molecular mechanisms of topical anti-inflammatory effects of lipoxin A(4) in endotoxin-induced uveitis. AB - Lipoxin A(4) (LXA(4)) is a lipid mediator that plays an important role in inflammation resolution. We assessed the anti-inflammatory effect of LXA(4) on endotoxin-induced uveitis (EIU) in rats. The inflammatory cell number and levels of tumor necrosis factor-alpha (TNF-alpha), interleukin-1beta (IL-1beta), prostaglandin E(2) (PGE(2)), and protein, as well as expression of cyclooxygenase 2 (COX-2) and vascular endothelial growth factor (VEGF), in the anterior chamber of the eye were determined 24 h after lipopolysaccharide (LPS; 200 mug/paw) intradermal injection. The immunohistochemical reactivities of nuclear factor kappaB (NF-kappaB) and c-Jun were also examined. Topical LXA(4) (1-10 ng/eye) pretreatment decreased the number of inflammatory cells and the protein leakage into the aqueous humor (AqH). In addition, topical LXA(4) (10 ng/eye) inhibited the LPS-induced production of IL-1beta, TNF-alpha, and PGE(2), and expression of COX-2 and VEGF. A decreased activation of NF-kappaB and c-Jun was also found in LXA(4)-treated eyes. It is very interesting that an anti-inflammatory effect was achieved even when LXA(4) (10 ng/eye) was applied topically after LPS challenge, as indicated by the reduction in the cellular and protein extravasations into the AqH. Moreover, topical treatment of corticosteroid prednisolone (200 mug/eye) beginning before or after LPS injection reduced all of the molecular and biochemical alterations promoted on EIU rats in an efficacy similar to that of LXA(4). Together, the present results provide clear evidence that pharmacological activation of LXA(4) signaling pathway potently reduces the EIU in rats. Therefore, LXA(4) stable analogs could represent promising agents for the management of ocular inflammatory diseases. PMID- 18413659 TI - Targeted disruption of murine organic anion-transporting polypeptide 1b2 (Oatp1b2/Slco1b2) significantly alters disposition of prototypical drug substrates pravastatin and rifampin. AB - Organic anion-transporting polypeptides (OATP) 1B1 and 1B3 are widely acknowledged as important and rate-limiting to the hepatic uptake of many drugs in clinical use. Accordingly, to better understand the in vivo relevance of OATP1B transporters, targeted disruption of murine Slco1b2 gene was carried out. It is noteworthy that Slco1b2(-/-) mice were fertile, developed normally, and exhibited no overt phenotypic abnormalities. We confirmed the loss of Oatp1b2 expression in liver using real-time polymerase chain reaction, Western Blot analysis, and immunohistochemistry. Expression of Oatp1a4 and Oatp2b1 but not Oatp1a1 was greater in female Slco1b2(-/-) mice, but expression of other non-OATP transporters did not significantly differ between wild-type and Slco1b2(-/-) male mice. Total bilirubin level was elevated by 2-fold in the Slco1b2(-/-) mice despite the fact that liver enzymes ALT and AST were normal. Pharmacological characterization was carried out using two prototypical substrates of human OATP1B1 and -1B3, rifampin and pravastatin. After a single intravenous dose of rifampin (1 mg/kg), a 1.7-fold increase in plasma area under the concentration time curve (AUC) was observed, whereas the liver-to-plasma ratio was reduced by 5 fold, and nearly 8-fold when assessed at steady-state conditions after 24 h of continuous subcutaneous infusion in Slco1b2(-/-) mice. Likewise, continuous subcutaneous infusion at low (8 microg/h) or high (32 microg/h) dose rates of pravastatin resulted in a 4-fold lower liver-plasma ratio in the in Slco1b2(-/-) mice. This is the first report of altered drug disposition profile in the Slco1b2 knockout mice and suggests the utility of this model for understanding the in vivo role of hepatic OATP transporters in drug disposition. PMID- 18413660 TI - Inhibition of Nod2 signaling and target gene expression by curcumin. AB - Nod2 is an intracellular pattern recognition receptor that detects a conserved moiety of bacterial peptidoglycan and subsequently activates proinflammatory signaling pathways. Mutations in Nod2 have been implicated to be linked to inflammatory granulomatous disorders, such as Crohn's disease and Blau syndrome. Many phytochemicals possess anti-inflammatory properties. However, it is not known whether any of these phytochemicals might modulate Nod2-mediated immune responses and thus might be of therapeutic value for the intervention of these inflammatory diseases. In this report, we demonstrate that curcumin, a polyphenol found in the plant Curcuma longa, and parthenolide, a sesquiterpene lactone, suppress both ligand-induced and lauric acid-induced Nod2 signaling, leading to the suppression of nuclear factor-kappaB activation and target gene interleukin-8 expression. We provide molecular and biochemical evidence that the suppression is mediated through the inhibition of Nod2 oligomerization and subsequent inhibition of downstream signaling. These results demonstrate for the first time that curcumin and parthenolide can directly inhibit Nod2-mediated signaling pathways at the receptor level and suggest that Nod2-mediated inflammatory responses can be modulated by these phytochemicals. It remains to be determined whether these phytochemicals possess protective or therapeutic efficacy against Nod2-mediated inflammatory disorders. PMID- 18413661 TI - Fluvastatin synergistically improves the antiproliferative effect of everolimus on rat smooth muscle cells by altering p27Kip1/cyclin E expression. AB - Multiple intracellular signaling pathways stimulate quiescent smooth muscle cells (SMCs) to exit from G(0) and re-enter the cell cycle. Thus, a combination of two drugs with different mechanisms of action may represent a suitable approach to control SMC proliferation, a prominent feature of in-stent restenosis. In the present study, we investigated the effect of everolimus, a mammalian target of rapamycin inhibitor, in combination with fluvastatin, a 3-hydroxy-3 methylglutaryl coenzyme A reductase inhibitor, on proliferation of rat SMCs. The antiproliferative action of everolimus was amplified by 2.5-fold by the addition of subliminal concentrations of fluvastatin (5 x 10(-7) M), lowering the IC(50) value from 2.5 x 10(-9) to 1.0 x 10(-9) M. The increased antiproliferative effect of everolimus by fluvastatin was prevented in the presence of mevalonate, farnesol, or geranylgeraniol, suggesting the involvement of prenylated proteins. Cell cycle analysis and [3H]thymidine incorporation assay demonstrated that the two drugs synergistically interfered with the progression of G(1) phase. In particular, the drug combination significantly up-regulated p27(Kip1) levels by 47.0%, suppressed cyclin E by 43.0%, and it reduced retinoblastoma (Rb) hyperphosphorylation by 79.0%, compared with everolimus alone. Retroviral overexpression of cyclin E conferred a significant resistance of rat SMCs to the antiproliferative action of the drug combination, measured by cell counting, [3H]thymidine incorporation, and cell cycle analysis, with higher levels of hyperphosphorylated form of Rb. Taken together, these results demonstrated that everolimus acts synergistically with fluvastatin to inhibit SMC proliferation by altering the expression of cyclin E and p27(kip1), which affects Rb phosphorylation and leads to G(1) phase arrest. PMID- 18413662 TI - Arrestin binds to different phosphorylated regions of the thyrotropin-releasing hormone receptor with distinct functional consequences. AB - Arrestin binding to agonist-occupied phosphorylated G protein-coupled receptors typically increases the affinity of agonist binding, increases resistance of receptor-bound agonist to removal with high acid/salt buffer, and leads to receptor desensitization and internalization. We tested whether thyrotropin releasing hormone (TRH) receptors lacking phosphosites in the C-terminal tail could form stable and functional complexes with arrestin. Fibroblasts from mice lacking arrestins 2 and 3 were used to distinguish between arrestin-dependent and -independent effects. Arrestin did not promote internalization or desensitization of a receptor that had key Ser/Thr phosphosites mutated to Ala (4Ala receptor). Nevertheless, arrestin greatly increased acid/salt resistance and the affinity of 4Ala receptor for TRH. Truncation of 4Ala receptor just distal to the key phosphosites (4AlaStop receptor) abolished arrestin-dependent acid/salt resistance but not the effect of arrestin on agonist affinity. Arrestin formed stable complexes with activated wild-type and 4Ala receptors but not with 4AlaStop receptor, as measured by translocation of arrestin-green fluorescent protein to the plasma membrane or chemical cross-linking. An arrestin mutant that does not interact with clathrin and AP2 did not internalize receptor but still promoted high affinity TRH binding, acid/salt resistance, and desensitization. A sterically restricted arrestin mutant did not cause receptor internalization or desensitization but did promote acid/salt resistance and high agonist affinity. The results demonstrate that arrestin binds to proximal or distal phosphosites in the receptor tail. Arrestin binding at either site causes increased agonist affinity and acid/salt resistance, but only the proximal phosphosites evoke the necessary conformational changes in arrestin for receptor desensitization and internalization. PMID- 18413663 TI - Atrial natriuretic peptide protects against histamine-induced endothelial barrier dysfunction in vivo. AB - Endothelial barrier dysfunction is a hallmark of many severe pathologies, including sepsis or atherosclerosis. The cardiovascular hormone atrial natriuretic peptide (ANP) has increasingly been suggested to counteract endothelial leakage. Surprisingly, the precise in vivo relevance of these observations has never been evaluated. Thus, we aimed to clarify this issue and, moreover, to identify the permeability-controlling subcellular systems that are targeted by ANP. Histamine was used as important pro-inflammatory, permeability increasing stimulus. Measurements of fluorescein isothiocyanate (FITC)-dextran extravasation from venules of the mouse cremaster muscle and rat hematocrit values were performed to judge changes of endothelial permeability in vivo. It is noteworthy that ANP strongly reduced the histamine-evoked endothelial barrier dysfunction in vivo. In vitro, ANP blocked the breakdown of transendothelial electrical resistance (TEER) induced by histamine. Moreover, as judged by immunocytochemistry and Western blot analysis, ANP inhibited changes of vascular endothelial (VE)-cadherin, beta-catenin, and p120(ctn) morphology; VE-cadherin and myosin light chain 2 (MLC2) phosphorylation; and F-actin stress fiber formation. These changes seem to be predominantly mediated by the natriuretic peptide receptor (NPR)-A, but not by NPR-C. In summary, we revealed ANP as a potent endothelial barrier protecting agent in vivo and identified adherens junctions and the contractile apparatus as subcellular systems targeted by ANP. Thus, our study highlights ANP as an interesting pharmacological compound opening new therapeutic options for preventing endothelial leakage. PMID- 18413664 TI - Formaldehyde-activated Pixantrone is a monofunctional DNA alkylator that binds selectively to CpG and CpA doublets. AB - The topoisomerase II poison mitoxantrone is important in the clinical management of human malignancies. Pixantrone, a novel aza-anthracenedione developed to improve the therapeutic profile of mitoxantrone, can efficiently alkylate DNA after formaldehyde activation. In vitro transcriptional analysis has now established that formaldehyde-activated pixantrone generates covalent adducts selectively at discrete CpG or CpA dinucleotides, suggesting that the activated complex binds to guanine or cytosine (or both) bases. The stability of pixantrone adduct-induced transcriptional blockages varied considerably, reflecting a mixture of distinct pixantrone adduct types that may include relatively labile monoadducts and more stable interstrand cross-links. 6,9-Bis-[[2 (dimethylamino)ethyl]amino]benzo[g]isoquinoline-5,10-dione (BBR 2378), the dimethyl N-substituted analog of pixantrone, could not form adducts, suggesting that pixantrone alkylates DNA through the primary amino functions located in each side chain of the drug. Pixantrone generated DNA adducts only when guanine was present in substrates and exhibited a lack of adduct formation with inosine containing polynucleotides, confirming that the N2 amino group of guanine is the site for covalent attachment of the drug. Mass spectrometric analysis of oligonucleotide-drug complexes confirmed that formation of covalent pixantrone DNA adducts is mediated by a single methylene linkage provided by formaldehyde and that this occurs only with guanine-containing double stranded oligonucleotide substrates. CpG methylation, an epigenetic modification of the mammalian genome, significantly enhanced the generation of pixantrone-DNA adducts within a methylated DNA substrate, indicating that the methylated dinucleotide may be a favored target in a cellular environment. PMID- 18413665 TI - AKT is activated in an ataxia-telangiectasia and Rad3-related-dependent manner in response to temozolomide and confers protection against drug-induced cell growth inhibition. AB - The phosphatidylinositol 3-kinase/AKT pathway is activated frequently in human cancer, and it has been implicated in tumor cell proliferation, survival, and chemoresistance. In this study, we addressed the role of AKT in cellular responses to the therapeutic methylating agent temozolomide (TMZ), and we investigated the possible link between TMZ-induced modulation of AKT function and activation of ataxia-telangiectasia and Rad3-related (ATR)- and ataxia telangiectasia mutated (ATM)-dependent signaling pathways. We found that clinically relevant concentrations of TMZ caused activation of endogenous AKT in lymphoblastoid cells, and in colon and breast cancer cells, and that this molecular event required a functional mismatch repair system. Transfection of a dominant-negative kinase-dead form of AKT1 into breast cancer cells abrogated TMZ induced activation of endogenous AKT, and it markedly enhanced cell sensitivity to the drug. Likewise, exposure of the MMR-proficient cell lines to the AKT inhibitor D-3-deoxy-2-O-methyl-myo inositol 1-[(R)-2-methoxy-3-(octadecyloxy) propyl hydrogen phosphate] (SH-5) impaired AKT phosphorylation in response to TMZ, and it significantly increased cell chemosensitivity. Furthermore, small interfering RNA (siRNA)-mediated reduction of AKT1 expression in colon cancer cells potentiated the growth inhibitory effects of TMZ. Inhibition of ATM expression in colon cancer cells by siRNA did not impair TMZ-induced activation of AKT, whereas siRNA-mediated inhibition of ATR prevented AKT activation in response to the drug and increased cell chemosensitivity. These results strongly support the hypothesis that clinical benefit could be obtained by combining TMZ with inhibitors of the AKT pathway. Moreover, they provide the first evidence of a novel function of ATR as an upstream activator of AKT in response to DNA damage induced by O(6)-guanine-methylating agents. PMID- 18413666 TI - Characterization of mENT1Delta11, a novel alternative splice variant of the mouse equilibrative nucleoside transporter 1. AB - Mammalian cells require specific transport mechanisms for the cellular uptake and release of endogenous nucleosides such as adenosine, and nucleoside analogs used in chemotherapy. We have identified a novel splice variant of the mouse equilibrative nucleoside transporter, mENT1, that results from the exclusion of exon 11 during pre-RNA processing. This variant encodes a truncated protein (mENT1Delta11) missing the last three transmembrane domains of the full-length mENT1. The mENT1Delta11 transcript and protein were found to be differentially distributed among tissues relative to full-length mENT1. PK15-NTD (nucleoside transport deficient) cells were transfected with mENT1 or mENT1Delta11 and assessed for nucleoside transport function. No significant differences were observed between the mENT1 and mENT1Delta11 in terms of transport function or inhibitor binding affinity. PK15-mENT1Delta11 transfected cells bound the ENT1 probe [3H]nitrobenzylthioinosine (NBMPR) with high affinity and mediated the cellular accumulation of both [3H]2-chloroadenosine and [3H]uridine. The only significant differences between the mENT1 variants were that mENT1Delta11 could not be photolabeled with [3H]NBMPR and that mENT1Delta11 was insensitive to the transporter-modifying effects of N-ethylmaleimide. These data suggest that the last three transmembrane domains of mENT1 are not necessary for transport activity, but this region does contain the cysteines responsible for the sensitivity of mENT1 to sulfhydryl reagents, and the residues important for covalent modification of the protein with NBMPR. These results provide important guidelines for future mutagenesis studies aimed at elucidating the tertiary structure of the ENT1 protein and the domains involved in inhibitor binding and substrate translocation. PMID- 18413668 TI - Patient--provider communication: understanding the role of patient activation for Latinos in mental health treatment. AB - This article highlights results from the Right Question Project-Mental Health (RQP-MH), an intervention designed to teach skills in question formulation and to increase patients' participation in decisions about mental health treatment. Of participants in the RQP-MH intervention, 83% were from a Latino background, and 75% of the interviews were conducted in Spanish. The authors present the steps participants undertook in the process of becoming "activated" to formulate effective questions and develop decision-making skills in relation to their care. Findings suggest that patient activation and empowerment are interdependent because many of the skills (i.e., question formulation, direct patient-provider communication) required to become an "activated patient" are essential to achieve empowerment. Also, findings suggest that cultural and contextual factors can influence the experience of Latinos regarding participation in health care interactions. The authors provide recommendations for continued research on the patient activation process and further application of this strategy in the mental health field, especially with Latinos. PMID- 18413669 TI - Desensitization of gonadotropin responses to kisspeptin in the female rat: analyses of LH and FSH secretion at different developmental and metabolic states. AB - Kisspeptins have emerged as potent elicitors of gonadotropin secretion and, therefore, putative targets for pharmacological intervention. In this context, desensitization of gonadotropin responses to continuous administration of kisspeptins has begun to be characterized, but information so far available is mostly restricted to LH responses in males, whereas the similar phenomenon in females, of obvious therapeutic interest, remains virtually unexplored. We report herein LH and FSH responses to continuous intracerebral administration of kisspeptin in female rats at different developmental and metabolic states. Infusion of kisspeptin-10 to adult female rats induced a transient elevation in serum LH concentrations, followed by a precipitous drop and normalization of LH levels thereafter. Elevation of LH after kisspeptin infusion was prolonged in underfed animals; a phenomenon mimicked by leptin administration. Conversely, FSH levels were persistently heightened along continuous kisspeptin infusion, but duration of this response was shortened by undernutrition. In pubertal females, LH and FSH levels remained elevated at the end of a 7-day infusion of kisspeptin; responses whose magnitude was augmented by subnutrition but not mimicked by leptin. In all settings, terminal gonadotropin-releasing hormone responses were fully preserved, suggesting that eventual desensitization must occur upstream from the pituitary. In summary, our current data document the pharmacological consequences of continuous administration of kisspeptin to female rats, with remarkable differences being detected between LH and FSH responses, in different developmental and metabolic states. These observations of potential pharmacological interest might help also to delineate the physiological roles of kisspeptins in the dynamic regulation of gonadotropin secretion in the female. PMID- 18413670 TI - Effects of c-MYC activation on glucose stimulus-secretion coupling events in mouse pancreatic islets. AB - Alteration of pancreatic beta-cell survival and Preproinsulin gene expression by prolonged hyperglycemia may result from increased c-MYC expression. However, it is unclear whether c-MYC effects on beta-cell function are compatible with its proposed role in glucotoxicity. We therefore tested the effects of short-term c MYC activation on key beta-cell stimulus-secretion coupling events in islets isolated from mice expressing a tamoxifen-switchable form of c-MYC in beta-cells (MycER) and their wild-type littermates. Tamoxifen treatment of wild-type islets did not affect their cell survival, Preproinsulin gene expression, and glucose stimulus-secretion coupling. In contrast, tamoxifen-mediated c-MYC activation for 2-3 days triggered cell apoptosis and decreased Preproinsulin gene expression in MycER islets. These effects were accompanied by mitochondrial membrane hyperpolarization at all glucose concentrations, a higher resting intracellular calcium concentration ([Ca(2+)](i)), and lower glucose-induced [Ca(2+)](i) rise and islet insulin content, leading to a strong reduction of glucose-induced insulin secretion. Compared with these effects, 1-wk culture in 30 mmol/l glucose increased the islet sensitivity to glucose stimulation without reducing the maximal glucose effectiveness or the insulin content. In contrast, overnight exposure to a low H(2)O(2) concentration increased the islet resting [Ca(2+)](i) and reduced the amplitude of the maximal glucose response as in tamoxifen-treated MycER islets. In conclusion, c-MYC activation rapidly stimulates apoptosis, reduces Preproinsulin gene expression and insulin content, and triggers functional alterations of beta-cells that are better mimicked by overnight exposure to a low H(2)O(2) concentration than by prolonged culture in high glucose. PMID- 18413671 TI - Effect of ovarian suppression with gonadotropin-releasing hormone agonist on glucose disposal and insulin secretion. AB - Several lines of evidence suggest that ovarian hormones influence glucose homeostasis, although their exact role in humans has not been clearly defined. In the present study, we sought to test the hypothesis that ovarian hormones regulate glucose homeostasis by examining the effect of pharmacologically induced ovarian hormone deficiency on glucose disposal and insulin secretion. Young, healthy women with regular menstrual patterns were studied during the follicular and luteal phases of their cycle at baseline and after 2 mo of treatment with gonadotropin-releasing hormone agonist (GnRHa; n = 7) or placebo (n = 6). Using hyperglycemic clamps, in combination with stable isotope-labeled (i.e., (13)C and (2)H) glucose tracers, we measured glucose disposal and insulin secretion. Additionally, we assessed body composition and regional fat distribution using radiologic imaging techniques as well as glucoregulatory hormones. Ovarian hormone suppression with GnRHa did not alter body composition, abdominal fat distribution, or thigh tissue composition. There was no effect of ovarian suppression on total, oxidative, or nonoxidative glucose disposal expressed relative to plasma insulin level. Similarly, no effect of ovarian hormone deficiency was observed on first- or second-phase insulin secretion or insulin clearance. Finally, ovarian hormone deficiency was associated with an increase in circulating adiponectin levels but no change in leptin concentration. Our findings suggest that a brief period of ovarian hormone deficiency in young, healthy, eugonadal women does not alter glucose disposal index or insulin secretion, supporting the conclusion that ovarian hormones play a minimal role in regulating glucose homeostasis. Our data do, however, support a role for ovarian hormones in the regulation of plasma adiponectin levels. PMID- 18413672 TI - Dimethyl amiloride improves glucose homeostasis in mouse models of type 2 diabetes. AB - Dimethyl amiloride (DMA) enhances insulin secretion in the pancreatic beta-cell. DMA also enhances time-dependent potentiation (TDP) and enables TDP to occur in situations where it is normally absent. As we have demonstrated before, these effects are mediated in part through inhibition of neuronal nitric oxide synthase (nNOS), resulting in increased availability of arginine. Thus both DMA and arginine have the potential to correct the secretory defect in diabetes by enabling or enhancing TDP. In the current study we have demonstrated the ability of these agents to improve blood glucose homeostasis in three mouse models of type 2 diabetes. The pattern of TDP under different conditions indicates that inhibition of NOS is not the only mechanism through which DMA exerts its positive effects. Thus we also have explored another possible mechanism through which DMA enables/enhances TDP, via the activation of mitochondrial alpha-ketoglutarate dehydrogenase. PMID- 18413673 TI - Characterization of a prospective human model for study of the reproductive hormone responses to major illness. AB - With critical illness, serum testosterone levels fall markedly, whereas estrogen levels rise. Although animal studies suggest adaptive advantages, no prospective model has been available for studies in humans. We hypothesized that coronary artery bypass graft (CABG) surgery would provide such a model by eliciting the same reproductive hormone and other endocrine responses as reported with major nonsurgical illnesses. We further hypothesized that those responses would occur consistently in all CABG patients with predictable time courses, providing reliable windows for prospective studies. In 17 men undergoing CABG, serum levels of reproductive hormones, cortisol, thyroid hormones, and IGF-I were measured before and for up to 5 wk after surgery. Changes in serum levels of reproductive and other hormones were similar to those reported in nonsurgical critically ill patients. Time course for onset, duration, and recovery of reproductive hormone changes were consistent among all patients. A window for studying the testosterone and estrogen responses was established as the first 5 days following CABG. Practical use of this model was demonstrated by evaluating, in another seven men, changes in gonadotroph responsiveness to GnRH following CABG. Finally, to determine whether our findings in CABG could be extended to other surgeries, we demonstrated similar endocrine responses in 12 men following abdominal aortic aneurysm resection. We conclude that patients undergoing CABG surgery provide a useful human model for the prospective evaluation of the reproductive axis responses to acute illness. Other major surgeries are likely to also be suitable for these studies. PMID- 18413674 TI - Regulation of cardiomyocyte hypertrophy in diabetes at the transcriptional level. AB - Diabetic cardiomyopathy, structurally characterized by cardiomyocyte hypertrophy and increased extracellular matrix (ECM) protein deposition, eventually leads to heart failure. We investigated the role of transcriptional coactivator p300 and its interaction with myocyte enhancer factor 2 (MEF2) in diabetes-induced cardiomyocyte hypertrophy. Neonatal rat cardiomyocytes were exposed to variable levels of glucose. Cardiomyocytes were analyzed with respect to their size. mRNA expression of p300, MEF2A, MEF2C, atrial natriuretic polypeptide (ANP), brain natriuretic polypeptide (BNP), angiotensinogen (ANG), cAMP-responsive element binding protein-binding protein (CBP), and protein analysis of MEF2 were done with or without p300 blockade. We investigated the hearts of STZ-induced diabetic rats and compared them with age- and sex-matched controls after 1 and 4 mo of followup with or without treatment with p300 blocker curcumin. The results were that cardiomyocytes, exposed to 25 mM glucose for 48 h, showed cellular hypertrophy and augmented mRNA expression of ANP, BNP, and ANG, molecular markers of cardiac hypertrophy. Glucose caused a duration-dependent increase of mRNA and protein expression in MEF2A and MEF2C and transcriptional coactivator p300. Curcumin, a p300 blocker, and p300 siRNA prevented these abnormalities. Similarly, ANP, BNP, and ANG mRNA expression was significantly higher in the hearts of diabetic rats compared with the controls, in association with increased p300, MEF2A, and MEF2C expression. Treatment with p300 blocker curcumin prevented diabetes-induced upregulation of these transcripts. We concluded that data from these studies demonstrate a novel glucose-induced epigenetic mechanism regulating gene expression and cardiomyocyte hypertrophy in diabetes. PMID- 18413675 TI - Cardiac overexpression of hormone-sensitive lipase inhibits myocardial steatosis and fibrosis in streptozotocin diabetic mice. AB - Intracellular lipid accumulation (steatosis) and resultant lipotoxicity are key features of diabetic cardiomyopathy. Since cardiac hormone-sensitive lipase (HSL) is activated in diabetic mice, we sought to explore a pathophysiological function of cardiac HSL in the development of diabetic cardiomyopathy. Transgenic (Tg) mice with heart-specific HSL overexpression were generated, and cardiac histology, function, lipid profile, and gene expressions were analyzed after induction of diabetes by streptozotocin. Electron microscopy showed numerous lipid droplets in wild-type (Wt) hearts after 3 wk of diabetes, whereas Tg mice showed no lipid droplet accumulation. Cardiac content of acylglycerides was increased approximately 50% with diabetes in Wt mice, whereas this was blunted in Tg hearts. Cardiac lipid peroxide content was twofold lower in Tg hearts than in Wt hearts. The mRNA expressions for peroxisome proliferator-activated receptor alpha, genes for triacylglycerol synthesis, and lipoprotein lipase were increased with diabetes in Wt hearts, whereas this induction was absent in Tg hearts. Expression of genes associated with lipoapoptosis was decreased, whereas antioxidant protein metallothioneins were increased in diabetic Tg hearts. Diabetic Wt hearts showed interstitial fibrosis and increased collagen content. However, Tg hearts displayed no overt fibrosis, concomitant with decreased expression of collagens, transforming growth factor-beta, and matrix metalloproteinase 2. Notably, mortality during the experimental period was approximately twofold lower in diabetic Tg mice compared with Wt mice. In conclusion, since HSL overexpression inhibits cardiac steatosis and fibrosis by apparently hydrolyzing toxic lipid metabolites, cardiac HSL could be a therapeutic target for regulating diabetic cardiomyopathy. PMID- 18413676 TI - Effect of GH on human skeletal muscle lipid metabolism in GH deficiency. AB - Adult-onset growth hormone (GH) deficiency (GHD) is associated with insulin resistance and decreased exercise capacity. Intramyocellular lipids (IMCL) depend on training status, diet, and insulin sensitivity. Using magnetic resonance spectroscopy, we studied IMCL content following physical activity (IMCL-depleted) and high-fat diet (IMCL-repleted) in 15 patients with GHD before and after 4 mo of GH replacement therapy (GHRT) and in 11 healthy control subjects. Measurements of insulin resistance and exercise capacity were performed and skeletal muscle biopsies were carried out to assess expression of mRNA of key enzymes involved in skeletal muscle lipid metabolism by real-time PCR and ultrastructure by electron microscopy. Compared with control subjects, patients with GHD showed significantly higher difference between IMCL-depleted and IMCL-repleted. GHRT resulted in an increase in skeletal muscle mRNA expression of IGF-I, hormone sensitive lipase, and a tendency for an increase in fatty acid binding protein-3. Electron microscopy examination did not reveal significant differences after GHRT. In conclusion, variation of IMCL may be increased in patients with GHD compared with healthy control subjects. Qualitative changes within the skeletal muscle (i.e., an increase in free fatty acids availability from systemic and/or local sources) may contribute to the increase in insulin resistance and possibly to the improvement of exercise capacity after GHRT. The upregulation of IGF-I mRNA suggests a paracrine/autocrine role of IGF-I on skeletal muscle. PMID- 18413677 TI - Pair feeding-mediated changes in metabolism: stress response and pathophysiology in insulin-resistant, atherosclerosis-prone JCR:LA-cp rats. AB - Rats of the JCR:LA-cp strain, which are homozygous for the cp gene (cp/cp), are obese, insulin-resistant, and hyperinsulinemic. They exhibit associated micro- and macrovascular disease and end-stage ischemic myocardial lesions and are highly stress sensitive. We subjected male cp/cp rats to pair feeding (providing the rats each day with the amount of food eaten by matched freely fed animals), a procedure that alters the diurnal feeding pattern, leading to a state of intermittent caloric restriction. Effects on insulin, glucose, and lipid metabolism, response to restraint stress, aortic contractile/relaxant response, and myocardial lesion frequency were investigated. Pair-fed young (12-wk-old) cp/cp rats had lower insulin and glucose levels (basal and following restraint), consistent with increased insulin sensitivity, but a greater increase in plasma nonesterified fatty acids in response to restraint. These effects were unrelated to lipolytic rates in adipose tissue but may be related to reduced fatty acid oxidation in skeletal muscle. Older (24-wk-old) pair-fed cp/cp rats had significantly reduced plasma triglyceride levels, improved micro- and macrovascular function, and reduced severity of ischemic myocardial lesions. These changes indicate a significant amelioration of end-stage disease processes in this animal model and the complexity of metabolic/physiological responses in studies involving alterations in food intake. The effects illustrate the sensitivity of the JCR:LA-cp rat, an animal model for the metabolic syndrome and associated cardiovascular disease, to the environmental and experimental milieu. Similar stress-related mechanisms may play a role in metabolically induced cardiovascular disease in susceptible human beings. PMID- 18413678 TI - Enhancement of tendon-bone osteointegration of anterior cruciate ligament graft using granulocyte colony-stimulating factor. AB - BACKGROUND: Whereas anterior cruciate ligament rupture usually requires reconstruction, the attachment between the tendon and the bone is the weakest region in the early posttransplantation period. In this process, the acquisition of appropriate vascularity is a key for early bone-tendon healing. HYPOTHESIS: Granulocyte colony-stimulating factor has an effect on the maturation of bone tendon integration of anterior cruciate ligament reconstruction. STUDY DESIGN: Controlled laboratory study. METHODS: Twenty-eight healthy adult beagle dogs underwent bilateral anterior cruciate ligament reconstruction using the ipsilateral flexor digitorum superficialis tendon and were divided into 2 groups. A granulocyte colony-stimulating factor-incorporated gelatin surrounded the graft in the granulocyte colony-stimulating factor group, and the same gelatin without granulocyte colony-stimulating factor was used as the control group. Assessment was done at 2 and 4 weeks. RESULTS: Histological analysis at week 2 demonstrated that, in addition to more Sharpey fibers, microvessels were significantly enhanced in the granulocyte colony-stimulating factor group's grafts. Computed tomography at week 4 showed a significantly smaller tibial bone tunnel in the granulocyte colony-stimulating factor group. Real-time polymerase chain reaction revealed significantly elevated messenger ribonucleic acid expression levels of vascular endothelial growth factor and osteocalcin in the tibial bone tunnel and graft compared with controls. Furthermore, biomechanical testing of force during loading to ultimate failure at week 4 demonstrated a significant increase in strength in the granulocyte colony-stimulating factor group. CONCLUSION: This study demonstrated that a local application of granulocyte colony-stimulating factor-incorporated gelatin significantly accelerates bone-tendon interface strength via enhanced angiogenesis and osteogenesis. CLINICAL RELEVANCE: Granulocyte colony-stimulating factor has therapeutic potential in promoting an environment conductive to angiogenesis and osteogenesis in bone tunnels. PMID- 18413679 TI - High tibial osteotomy for unloading osteochondral defects in the medial compartment of the knee. AB - BACKGROUND: High tibial osteotomy is a well-established method for the treatment of symptomatic medial unicompartmental cartilage degeneration. While the findings of several outcome studies have led to the widely practiced postoperative goal alignment of 8 degrees to 10 degrees of valgus, there exists no literature to account for patient-specific measurements such as body weight, baseline geometry, and medial chondral defect size. Furthermore, there is a lack of literature to support the ideal goal alignment when using high tibial osteotomy to unload isolated chondral defects. PURPOSE: To identify a relationship between these patient-specific factors and the ideal postoperative alignment. STUDY DESIGN: Controlled laboratory study. METHODS: High tibial osteotomy was performed on 8 human cadaveric knees and was fixed with a dynamic external fixator. The fixator was used to vary the tibiofemoral alignment from 12 degrees valgus to 10 degrees varus. At each alignment, force, contact area, and pressure distribution were recorded. This pattern was repeated for a range of applied loads (0.8-1.8 x ideal body weight) and across a range of medial chondral defect sizes (10-20 cm). RESULTS: As tibiofemoral alignment was shifted from varus to valgus alignment, we found a decrease in medial contact pressure (P < .001) and a decrease in medial contact area (P < .001). For all defect sizes, all contact pressure within the medial compartment was shifted to the lateral compartment at between 6 degrees and 10 degrees of valgus. Contact pressure was found to concentrate around the defect rims for all defect sizes. CONCLUSION: We show that regardless of condylar width, baseline tibiofemoral alignment, body weight, or chondral defect size, all specimens demonstrated complete unloading of the medial compartment at between 6 degrees and 10 degrees of valgus, which favors cartilage repair at these alignments. In addition, regarding the use of high tibial osteotomy for unloading isolated chondral defects, we find that contact pressure is approximately equally distributed between the medial and lateral compartments for alignments of 0 degrees to 4 degrees of valgus. CLINICAL RELEVANCE: This loading situation most closely approximates physiologic loading and therefore represents an ideal outcome for patients with isolated chondral defects. Reduction in stress concentration around chondral defects has been found to favor cartilage repair. PMID- 18413680 TI - Comparison of the clinical outcomes of single- and double-row repairs in rotator cuff tears. AB - BACKGROUND: Although research has demonstrated the superiority of double-row rotator cuff repair over single-row methods from a biological and mechanical point of view, few studies have compared clinical outcome of the 2 methods, and no articles have been published describing the superiority of double-row methods in clinical aspects. HYPOTHESIS: Arthroscopic double-row repair of a rotator cuff tear has superior clinical outcome to single-row repair. STUDY DESIGN: Cohort study; Level of evidence, 2. METHODS: The study included 78 patients operated on for full-thickness rotator cuff tears between May 2002 and May 2004. A single-row fixation method was used in the first consecutive 40 patients, and a double-row fixation method was used in the next consecutive 38 patients. The mean age at surgery was 56 years. At 2 years after surgery, final evaluation was done with American Shoulder and Elbow Surgeons and Constant scoring systems and the Shoulder Strength Index. The Shoulder Strength Index is a new evaluation method to estimate relative shoulder strength compared with the unaffected shoulder. RESULTS: At final follow-up, the average American Shoulder and Elbow Surgeons scores were 91.6 in the single-row group and 93.0 in the double-row group. The Constant score was 76.7 in the single-row group and 80.0 in the double-row group. Functional outcome was improved in both groups after surgery, but there was no significant difference between the 2 groups. When the patients were further divided by size of tear, there was still no difference between the repair techniques in the patients with small to medium (<3 cm) tears. However, in patients with large to massive tears (>3 cm), the American Shoulder and Elbow Surgeons and Constant scores and Shoulder Strength Index were all significantly better in the group that had double-row repair. CONCLUSION: Small to medium rotator cuff tears should be repaired with the single-row method, and large to massive tears should be repaired with the double-row method. PMID- 18413681 TI - Rotator cuff degeneration: etiology and pathogenesis. AB - By virtue of its anatomy and function, the rotator cuff is vulnerable to considerable morbidity, often necessitating surgical intervention. How we intervene is governed by our understanding of the pathological mechanisms in cuff disease. These factors can be divided into those extrinsic to the rotator cuff (impingement, demographic factors) and those intrinsic to the cuff (age-related degeneration, hypovascularity, inflammation, and oxidative stress, among others). In an era where biologic interventions are increasingly being investigated, our understanding of these mechanisms is likely to become more important in designing effective new interventions. Here we present a literature review summarizing our current understanding of the pathophysiological mechanisms underlying rotator cuff degeneration. PMID- 18413682 TI - Treatment of postpartum depression with psychotic features in a patient with tropical spastic paraparesis. PMID- 18413683 TI - Efficacy of tamsulosin with extracorporeal shock wave lithotripsy for passage of renal and ureteral calculi. AB - OBJECTIVE: To review the evidence for the safety and efficacy of adjunctive tamsulosin in enhancing the efficacy of renal and ureteral stone clearance when used with extracorporeal shock wave lithotripsy (ESWL). DATA SOURCES: A search of MEDLINE (1950-January 2008), PubMed (1950-January 2008), and the Iowa Drug Information System (1966-January 2008) was performed using the search terms tamsulosin and extracorporeal shock wave lithotripsy. MeSH headings included lithotripsy and adrenergic alpha-antagonists. Additional references were found by searching bibliographic references of resulting citations. STUDY SELECTION AND DATA EXTRACTION: All studies utilizing tamsulosin therapy after a single session of ESWL or after the development of steinstrasse, an accumulation of stone fragments that obstructs the ureter, were included. DATA SYNTHESIS: To date, 5 prospective studies have evaluated the efficacy of tamsulosin combined with ESWL in enhancing the passage of renal and ureteral stones. In one trial, 12-week renal stone clearance was 60% in the control group compared with 78.5% in the tamsulosin group (p = 0.037). Among trials that evaluated overall ureteral stone clearance, efficacy rates were 33.3-79.3% in the control groups compared with 66.6-96.6% in the tamsulosin groups. Reports of pain and supplemental analgesic dosing were consistently lower with tamsulosin, but data on the incidence of subsequent retreatment with ESWL or ureteroscopy was rarely reported. Adjunctive tamsulosin particularly enhanced the passage of renal stones 10-24 millimeters in diameter. Overall, tamsulosin was well tolerated. CONCLUSIONS: Overall, evidence suggests that adjunctive tamsulosin therapy combined with ESWL is safe and effective in enhancing stone clearance in patients with renal stones 10-24 millimeters in diameter. Evidence regarding ureteral stone clearance is inconclusive, although adjunctive tamsulosin has been reported to reduce painful episodes. Larger prospective trials evaluating different dosages and stone locations, as well as the ability of tamsulosin to reduce repeat ESWL or more invasive methods such as ureteroscopy should be performed. PMID- 18413684 TI - Comparison of two point-of-care lipid analyzers for use in global cardiovascular risk assessments. AB - BACKGROUND: Point-of-care (POC) lipid testing is increasingly used in community- and office-based practice. Two analyzers commonly used in the US are CardioChek PA and Cholestech LDX. Both directly measure total cholesterol (TC) and high density lipoprotein cholesterol (HDL-C), mandatory values in calculating a Framingham Risk Score (FRS). The FRS in turn informs the clinician of the need for lipid-modifying therapy and the degree of therapeutic intensity. OBJECTIVE: To compare the performance of CardioChek PA and Cholestech LDX. METHODS: Staff members from the Colorado Prevention Center were included in the study, with all having fasted for 12 hours before the testing. No medical history was obtained. A venous blood sample was collected for lipid measurements conducted in a laboratory, and 2 finger sticks were obtained at that time and analyzed immediately on-site using the POC analyzers. Intraclass correlation coefficients (ICCs) were determined for each analyzer versus the laboratory analysis, with values greater than 0.75 defined as indicators of excellent reproducibility. We then assessed how interanalyzer differences in TC or HDL-C impacted the FRS lipid categorization. RESULTS: Thirty-four adults (aged 24-56 y) participated in the study. The ICC between Cholestech LDX and the laboratory standard exceeded 0.75 for all 4 lipid categories (TC, rho = 0.96; HDL-C, rho = 0.88; low-density lipoprotein cholesterol, rho = 0.87; triglycerides, rho = 0.99). By contrast, the only ICC exceeding 0.75 using CardioChek PA was for triglycerides (rho = 0.84). When applied in calculating the FRS, the Cholestech LDX analyzer misclassified fewer individuals for TC versus the CardioChek PA analyzer (5 vs 21). Overall, Cholestech LDX provided TC and HDL-C values in the correct FRS category more frequently versus CardioChek PA (TC, p < 0.001; HDL-C, p < 0.001). Limitations of the study include use of only 2 POC products and small sample size with no known risk factors. This project does not prove superior accuracy of either device, but reflects a real-world comparison of the analyzers conducted at a single center. CONCLUSIONS: The Cholestech LDX analyzer demonstrated better reproducibility than the CardioChek PA analyzer when compared with laboratory gold standard analysis and allowed more accurate categorization for FRS. Since results obtained from these analyzers have the potential to impact treatment decisions, larger, prospective, comparative studies seem warranted. PMID- 18413685 TI - Corticosteroids for prevention of postextubation laryngeal edema in adults. AB - OBJECTIVE: To evaluate the efficacy and safety of prophylactic corticosteroid therapy in preventing postextubation laryngeal edema (PELE) and the need for reintubation in adults. DATA SOURCES: Literature was accessed through MEDLINE (1966-January 2008) and the Cochrane Library using the terms laryngeal edema, airway obstruction, postextubation stridor, intubation, glucocorticoids, and corticosteroids. Bibliographies of cited references were reviewed and a manual search of abstracts from recent pulmonary and critical care meetings was completed. STUDY SELECTION AND DATA EXTRACTION: All English-language, placebo controlled, randomized studies evaluating the use of prophylactic corticosteroids for the prevention of postextubation laryngeal edema or postextubation stridor (PES) in adults were reviewed. DATA SYNTHESIS: Although laryngoscopy is the gold standard method for diagnosing PELE, PES is more commonly used for diagnosis in clinical practice. While 3 older studies failed to demonstrate benefit with the prophylactic administration of corticosteroid therapy in terms of reducing PELE, PES, or the need for reintubation, each of these studies evaluated only a single dose of steroid therapy that was initiated only 30-60 minutes prior to a planned extubation in a population of patients at low-risk for PELE. In comparison, 3 newer studies, each using 4 doses of corticosteroid therapy initiated 12-24 hours prior to a planned extubation in patients deemed to be at high baseline risk for developing PELE, demonstrated a reduction in PELE, PES, and the need for reintubation; no safety concerns were identified. Current evidence therefore suggests that prophylactic intravenous methylprednisolone therapy (20-40 mg every 4-6 h) should be considered 12-24 hours prior to a planned extubation in patients at high-risk for PELE (eg, mechanical ventilation > 6 days). CONCLUSIONS: Data from the most recent well-designed clinical trials suggest that prophylactic corticosteroid therapy can reduce the incidence of PELE and the subsequent need for reintubation in mechanically ventilated patients at high-risk for PELE. Based on this information, clinicians should consider initiating prophylactic corticosteroid therapy in this population. Further studies are needed to establish the optimal dosing regimens as well as the subgroups of patients at high risk for PELE who will derive the greatest benefit from this preventive steroid therapy. PMID- 18413686 TI - Rifampin-related acute renal failure, thrombocytopenia, and leukocytoclastic vasculitis. PMID- 18413687 TI - Interactions between warfarin and three commonly prescribed fluoroquinolones. AB - OBJECTIVE: To critically evaluate a possible increased anticoagulant response during concomitant warfarin and fluoroquinolone therapy. DATA SOURCES: A literature search was conducted using PubMed, International Pharmaceutical Abstracts, and MEDLINE, from inception to January 2008, combining the term warfarin individually with ciprofloxacin, levofloxacin, and moxifloxacin. These 3 quinolones were selected based on their commercial availability and use in the US. STUDY SELECTION AND DATA EXTRACTION: All publication types including human participants and published in English were eligible for review. Reports were selected based on the use of typical treatment courses of fluoroquinolones during concomitant warfarin therapy and the reporting of prothrombin time (PT) or international normalized ratio (INR). DATA SYNTHESIS: Twenty-two publications were evaluated including 16 case reports or case series, 2 retrospective cohort studies, and 4 prospective studies, which included 2 placebo-controlled investigations. Identified reports covered a wide range of patient ages with multiple comorbidities. Changes in PT and INR values were considerably variable and inconsistent during concomitant warfarin and fluoroquinolone therapy. Results from the 6 structured reports demonstrated mean increases in PT and INR values that were clinically insignificant. However, some patients experienced significant increases above the desired therapeutic range. Increased anticoagulation was typically observed within the first week of concomitant fluoroquinolone therapy. Bleeding complications during times of increased anticoagulation were not always observed, but did result in death for 2 patients. CONCLUSIONS: Published data show no consistent increase in anticoagulant effects during concomitant warfarin and 3 commonly prescribed fluoroquinolones. Therefore, more frequent monitoring during concomitant therapy would be prudent. PMID- 18413688 TI - Immunologic benefits of enfuvirtide in patients enrolled in a drug assistance program. AB - BACKGROUND: Randomized clinical trials have demonstrated that enfuvirtide plus an optimized background regimen can cause a significant increase in CD4+ cell counts and a reduction in HIV RNA levels. OBJECTIVE: To describe and analyze CD4+ cell count and HIV RNA changes in HIV-infected patients receiving enfuvirtide and a prescribed background regimen (PBR) in a primarily clinical setting. METHODS: A retrospective review from September 1998 through August 2005 of CD4+ cell counts and HIV RNA changes from baseline was conducted in patients receiving enfuvirtide. Data were stratified and analyzed according to baseline CD4+ cell count and HIV RNA. RESULTS: A mean CD4+ cell count increase of approximately 102 cells/mm(3)was observed, regardless of baseline CD4+ cell count, in 187 patients receiving enfuvirtide during a mean of 19.4 months of follow-up. During 3 years of follow-up, patients initiating enfuvirtide at CD4+ cell counts less than 100 cells/mm(3)never achieved absolute CD4+ cell counts comparable to the counts in patients starting enfuvirtide at CD4+ cell counts of 100 cells/mm(3)or more. In 38.3% of patients achieving an undetectable HIV RNA level, a mean CD4+ cell count increase of 185 cells/mm(3)was observed. An unexpected finding was that a mean CD4+ cell count increase of 76 cells/mm(3)occurred in 61.7% of patients not achieving complete viral suppression. CONCLUSIONS: Immunologic benefits were observed in subjects continuing enfuvirtide plus a PBR irrespective of baseline CD4+ cell count, complete viral suppression, or antiretroviral susceptibility data. Data suggest that initiation of enfuvirtide at CD4+ cell counts greater than 100 cells/mm(3)may be immunologically advantageous and independent of complete virologic response. PMID- 18413689 TI - Single antiplatelet therapy for patients with previous gastrointestinal bleeds. AB - OBJECTIVE: To determine whether aspirin plus a proton pump inhibitor (PPI) is preferable, from a gastrointestinal bleed (GIB) risk perspective, to clopidogrel in patients who have experienced a GIB while on aspirin and who require single antiplatelet therapy for secondary prevention of cardiovascular disease. DATA SOURCES: A literature search was conducted using EMBASE (1980-January 2008), PubMed (1966-January 2008), Google, and a manual search of the reference lists using the search terms gastrointestinal bleed, gastrointestinal hemorrhage, peptic ulcer hemorrhage, ASA, aspirin, Plavix, clopidogrel, and PPI. The search, limited to human and English studies, yielded 110 returns. STUDY SELECTION AND DATA EXTRACTION: Randomized trials that compared aspirin with clopidogrel, involved patients who had previously experienced a GIB, and provided detailed information on the type and dose of drugs used were included. Studies were required to provide information on the recurrence of GIB. DATA SYNTHESIS: Two randomized trials were reviewed to assess the safety of secondary prevention of cardiovascular disease with respect to previous GIB. These noninferiority trials compared aspirin plus a PPI with clopidogrel over 12 months following confirmed healing of an aspirin-induced ulcer. In both trials, the majority of the GIB recurrences were in the clopidogrel group (8.6% vs 0.7%; difference 7.9%; 95% CI 3.4 to 12.4; p = 0.001 and 13.6% vs 0%; difference 13.6%; 95% CI 6.3 to 20.9; p = 0.0019) and the difference in recurrence rates exceeded the a priori selected upper boundary. CONCLUSIONS: Findings reported in the limited literature available support that clopidogrel is not equivalent to the combination of aspirin plus a PPI in the patient population studied. Aspirin plus a PPI would be considered clinically superior and should be used in medically managed patients who require single antiplatelet therapy but have had a prior GIB while on aspirin. Further research regarding dual antiplatelet therapy and a PPI is required. PMID- 18413690 TI - Comparison of simvastatin tablets from the US and international markets obtained via the Internet. AB - BACKGROUND: Convenient access to prescription drugs produced outside the US has been facilitated by the Internet. Of greatest concern to clinicians and policymakers is product quality and patient safety. The Food and Drug Administration has issued warnings to potential buyers that the safety of drugs purchased through the Internet cannot be guaranteed and may present consumers with a health risk from substandard products. OBJECTIVE: To determine whether generic simvastatin tablets and capsules obtained via the Internet from international markets are equivalent to the US innovator product regarding major aspects of pharmaceutical quality. METHODS: Twenty simvastatin tablets and capsules were obtained for pharmaceutical analysis: 19 generic samples from international Internet pharmacy Web sites and the US innovator product. Tablet samples were tested according to US Pharmacopeial (USP) guidelines where applicable, using high-performance liquid chromatography, disintegration, dissolution, weight variation, hardness, and assessment of physical characteristics. These tests are often used to detect formulation defects of drug products during the manufacturing process. RESULTS: Several international samples analyzed were not comparable to the US product in one or more aspects of quality assurance testing, and significant variability was found among foreign-made tablets themselves. Five samples failed to meet USP standards for dissolution and 2 for content uniformity. Among all samples, variability was observed in hardness, weight, and physical characterization. CONCLUSIONS: Results suggest that manufacturing standards for the international generic drug products compared with the US innovator product are not equivalent with regard to quality attributes. These findings have implications for safety and effectiveness that should be considered by clinicians to potentially safeguard patients who choose to purchase foreign-manufactured drugs via the Internet. PMID- 18413691 TI - Complexity of interactions between voriconazole and antiretroviral agents. AB - OBJECTIVE: To evaluate and summarize pertinent aspects of the literature on interactions between voriconazole and antiretroviral agents. DATA SOURCES: Primary literature was identified through MEDLINE (1950-February 2008), EMBASE (1988-February 2008), and International Pharmaceutical Abstracts (1970-February 2008) using the search terms voriconazole, ritonavir, protease inhibitors, nonnucleoside reverse transcriptase inhibitors, raltegravir, maraviroc, and drug interactions. Additionally, relevant abstracts from infectious diseases and HIV conferences (2004-February 2008), reference citations from relevant publications, and product information monographs were reviewed. STUDY SELECTION AND DATA ABSTRACTION: All articles identified from the data sources and published in English were reviewed. Of these, studies and reports addressing voriconazole pharmacokinetics or interactions with antiretroviral agents were evaluated. DATA SYNTHESIS: The interactions between voriconazole and antiretroviral drugs are complex. Voriconazole and ritonavir exhibit a time- and dose-dependent interaction. Ritonavir initially inhibits voriconazole metabolism, but, with chronic administration, subsequently induces voriconazole metabolism. This interaction is more pronounced with high doses of ritonavir. Coadministration of voriconazole and efavirenz at usual doses is contraindicated because of a 2-way interaction resulting in efavirenz toxicity and decreased therapeutic effect of voriconazole. Dosage adjustments of both drugs are required. Based on pharmacokinetic characteristics, interactions between voriconazole and other protease inhibitors, nonnucleoside reverse transcriptase inhibitors (including etravirine), and maraviroc are likely but have not been well characterized in the literature. Interactions between voriconazole and nucleoside reverse transcriptase inhibitors or raltegravir are not anticipated. CONCLUSIONS: Interactions between voriconazole and antiretrovirals have the potential for serious consequences. However, because there is limited information available, further studies are warranted to establish these interactions and clarify their appropriate management. Until then, clinicians should be aware of the potential for interactions between voriconazole and antiretroviral agents and how to monitor for these interactions in clinical practice. PMID- 18413692 TI - Intravenous human plasma-derived augmentation therapy in alpha 1-antitrypsin deficiency: from pharmacokinetic analysis to individualizing therapy. AB - BACKGROUND: Severe forms of alpha(1)-antitrypsin (AAT) deficiency require augmentation therapy by intravenous administration of purified preparations of AAT concentrate. Although standard AAT treatment schedules are widely available, pharmacokinetic studies characterizing AAT serum decay are scarce, and data on the variability of individual patients are almost nonexistent. OBJECTIVE: To establish individual AAT pharmacokinetics and develop a predictive model based on simple pharmacokinetic characterization that can be used to optimize individual AAT dosing regimens. METHODS: Seven patients with severe hereditary AAT deficiency (PI(*)ZZ phenotype) with serum AAT levels less than 0.50 g/L initially received AAT 180 mg/kg every 3 weeks. At 7, 14, and 21 days after AAT administration, serum samples were taken for quantitative AAT analysis and further one-compartment pharmacokinetic analysis. Subsequently, patients were rescheduled (dose and dosing interval) according to their individual responses. The influence of baseline AAT level, age, sex, body weight, and commercial AAT preparation was evaluated. RESULTS: The mean +/- SD AAT pharmacokinetic profile was: volume of distribution 127.6 +/- 31.9 mL/kg, clearance 10.13 +/- 1.84 mL/kg/day, and half-life 8.7 +/- 1.0 days. Hence, the mean optimized final AAT dose was 123.1 mg/kg every 2 weeks (range 118.5-125.6). AAT concentrations differed by a mean (geometrical) value of 3.9% (range -4.2% to 6.7%) from the minimum desired AAT serum trough of 0.50 g/L. The impact of baseline AAT levels and commercial AAT preparation used was statistically significant (p = 0.033 and p = 0.035, respectively). Differences between estimated and actual values were slightly lower when baseline AAT levels were taken into consideration, with a mean value of 3.3% (range -4.2% to 6.1%). CONCLUSIONS: AAT augmentation therapy can be effectively individualized on a pharmacokinetic basis with a simple, easily executed method. PMID- 18413693 TI - Valid treatment options for osteoporosis and osteopenia in HIV-infected persons. AB - OBJECTIVE: To review clinical data on bone ossification agents that may be considered for use in the treatment of osteoporosis and osteopenia in HIV infected patients. DATA SOURCES: A literature search was performed using MEDLINE (1950-January 2008), EMBASE, PubMed, and abstracts from major HIV conferences (February 2001-October 2007). These searches were limited to human data published in English and used the key words bisphosphonates, calcitonin, raloxifene, teriparatide, HAART, osteopenia, osteoporosis, and HIV/AIDS. Additional articles were retrieved from citations of selected references. STUDY SELECTION AND DATA EXTRACTION: Relevant information on the pharmacology, pharmacokinetics, safety, and efficacy of available treatment with hormonal and nonhormonal agents was selected. Greater emphasis was placed on randomized clinical trials than on retrospective studies. DATA SYNTHESIS: Osteoporosis in HIV-infected persons is at least as prevalent as in postmenopausal women, yet this population is not listed in primary care guidelines as one that should be considered for screening. In addition to bisphosphonates, calcitonin, raloxifene, and teriparatide are used to treat bone disorders. Three clinical trials to date have evaluated the use of a bisphosphonate in HIV-infected persons. The trials showed a marked increase in bone mineral density in patients taking alendronate versus those in the control groups (with/without calcium, exercise, and/or vitamin D in 1 or both arms). Dosing restrictions complicate the use of these agents; diet, exercise, and calcium supplementation remain the foremost recommended strategies to prevent bone loss. The use of estrogen, testosterone, calcitonin, and teriparatide is less studied in HIV-positive patients, but may be considered in select cases. There are some investigational drugs and agents not available in the US; however, there are not enough data to support their use. CONCLUSIONS: Alendronate appears to be a promising treatment option for HIV-infected patients with osteoporosis and osteopenia. Further research is required to determine the safety and efficacy of other available drugs. Until additional information is provided, and with available knowledge on the metabolism profiles of antiretroviral and bone ossification agents, alendronate appears to be the preferred agent to use in this population. PMID- 18413694 TI - Challenges to evidence-based prescribing in clinical practice. AB - Although there appears to be widespread support of evidence-based medicine as a basis for rational prescribing, the challenges to it are significant and often justified. A multitude of factors other than evidence drive clinical decision making, including patient preferences and social circumstances, presence of disease-drug and drug-drug interactions, clinical experience, competing demands from more pressing clinical conditions, marketing and promotional activity, and system-level drug policies. PMID- 18413695 TI - Pharmacist-administered admission histories: focus on immunizations in medication reconciliation. PMID- 18413696 TI - Use of the observation option and compliance with guidelines in treatment of acute otitis media. PMID- 18413697 TI - Clinical profile of the overweight child in the new millennium. AB - BACKGROUND: In conjunction with the rising prevalence of obesity during the past several decades, the clinical profile of the obese child has changed. Hypothesis. Environmental influences and eating practices have had an impact on the presence of medical morbidities among obese children. DESIGN: Retrospective chart review of data collected from 90 children entering into a pediatric weight management program was performed. Fisher's exact tests and Wilcoxon rank sum tests were used to compare outcomes between subpopulations. RESULTS: There was greater elevation in systolic blood pressure among children who ate in front of the television (P = .03) and a greater degree of fast-food consumption among children with more than 3 medical morbidities (P = .02). Breast-feeding did not have a protective effect on the degree of obesity (P = .02). CONCLUSION: Aggressive assessment for symptoms should be an important part of evaluating the overweight child. Environmental influences and social feeding practices can counteract the protective effects of breast-feeding in infancy. PMID- 18413698 TI - A dimensional analysis of the concept: good death of a child with cancer. AB - The death of a child is painful to all witnesses, but the experience varies among individuals, and differing perspectives lead to a variety of definitions for a good death. This analysis is undertaken to clarify the dimensions involved in a good death of a child with cancer and to examine them from 3 perspectives: the dying child, the child's family, and the health care providers. PMID- 18413699 TI - Establishing literature-based items for an oral mucositis assessment tool in children. AB - Although there are numerous instruments for oral mucositis (OM) assessment in adults, there is a lack of validated instruments for use in children. The objectives of this systematic review were to describe literature-based items that should be considered for a pediatric mucositis scale and other issues that should be considered when assessing mucositis in children. Literature search of PubMed and bibliography searches identified articles relevant to the assessment of mucositis and mucositis assessment studies in children. The elements established from the literature were grouped under objective, subjective, and functional items. Other issues that require consideration include the conditions for the assessment of the oral cavity and the etiology of OM. The resultant list included 23 items that have been used in mucositis assessment scales. This list of items may be used as an initial step in developing a new pediatric OM scale that particularly focuses on the unique issues in children. PMID- 18413700 TI - Peer-supported storytelling for grieving pediatric oncology nurses. AB - Telling stories about deceased patients to supportive peers is frequently mentioned as an activity used for meaning-making in anecdotal reports of clinical practice and the literature addressing nurses' experiences caring for dying children. This study examines peer-supported storytelling for grieving pediatric oncology nurses using a mixed methods single-group descriptive repeated measures design. Participants were 6 registered nurses from a tertiary care pediatric hospital inpatient oncology unit who self-identified as experiencing grief. Participants met in self-selected dyads for 2 storytelling sessions. Questionnaires were completed at baseline, midpoint, and study end. Sessions were audio-recorded. Participants reported (1) receiving and providing support during sessions; (2) that sessions had an impact on their grief; (3) that sessions had an impact on their meaning-making, and the explicit session focus on making sense of and identifying benefit in their experiences was particularly helpful. There was a significant positive correlation between participant report of number of special patient deaths during career and impact of sessions on grief. PMID- 18413701 TI - Plasmodium falciparum-free merozoites and infected RBCs distinctly affect soluble CD40 ligand-mediated maturation of immature monocyte-derived dendritic cells. AB - Free plasmodium merozoites released from the parasitized hepatocytes and erythrocytes represent a transitory, extracellular stage in its mammalian host. In this study, we compared the effect of Plasmodium falciparum-free merozoites with infected RBCs (iRBCs) on the maturation of human monocyte-derived dendritic cells (DCs) in vitro. Phagocytosed-free merozoites prevented soluble CD40 ligand (sCD40L)-induced, phenotypic maturation of DCs and secretion of IL-12p70 but enhanced IL-10 production and primed, naive CD4+ cells to produce a high level of IL-10 compared with IFN-gamma. Free merozoites augmented sCD40L-induced ERK1/2 activation, and inhibition of ERK1/2 with its inhibitor PD98059 markedly abrogated IL-10 production and rescued IL-12 production. Therefore, the molecular mechanisms by which free merozoites antagonized sCD40L-induced DC maturation appeared to involve the activation of the ERK pathway. In contrast, phagocytosed iRBCs by itself induced DCs to semi-maturation, responded to CD40 signaling by maturing and secreting increased levels of TNF-alpha, IL-6, and also IL-12p70, and led to a pronounced, proinflammatory response by the allogenic CD4+ T cells. iRBCs regulate CD40-induced p38MAPK. Studies using inhibitors selective for p38MAPK (SB203580) showed that p38MAPK played an essential role in the maturation and function of DCs. Our results reveal the ability of free merozoites and iRBCs to distinctly alter the sCD40L-induced DC functioning by regulating the activation of the MAPK pathway that can inactivate or exacerbate immune responses to promote their survival and the development of parasite-specific pathologies. PMID- 18413702 TI - Gray matter structural alterations in psychotropic drug-naive pediatric obsessive compulsive disorder: an optimized voxel-based morphometry study. AB - OBJECTIVE: Although several magnetic resonance imaging (MRI) studies have been conducted in adults with obsessive-compulsive disorder (OCD), few studies have used voxel-based morphometry to examine brain structure, especially in psychotropic drug-naive pediatric patients. METHOD: MRI examinations of 37 psychotropic drug-naive pediatric OCD patients and 26 age- and sex-matched healthy volunteers were acquired on a 1.5 T MRI system, normalized to a customized template, and segmented with optimized voxel-based morphometry. RESULTS: Pediatric OCD patients had significantly more gray matter in regions predicted to differ a priori between groups, including the right and left putamen and orbital frontal cortex. Among patients, more gray matter in the left putamen and right lateral orbital frontal cortex correlated significantly with greater OCD symptom severity, but not with anxiety or depression. Manual region-of interest measurements confirmed more gray matter in the orbital frontal cortex and putamen in patients compared to healthy volunteers. More anterior cingulate gray matter was evident among patients compared to healthy volunteers with regional volumetry but not with voxel-based morphometry. Regions of significantly less gray matter in OCD were confined to the occipital cortex and were not predicted a priori. CONCLUSIONS: Our results suggest that OCD is characterized by more gray matter in brain regions comprising cortical-striatal-thalamic-cortical circuits. These findings are consistent with functional neuroimaging studies reporting hypermetabolism and increased regional cerebral blood flow in striatal, anterior cingulate, and orbital frontal regions among OCD patients while in a resting state. PMID- 18413703 TI - Cost-effectiveness of treatments for adolescent depression: results from TADS. AB - OBJECTIVE: While the evidence base for treatments for adolescent depression is building, little is known about the relative efficiency of such treatments. Treatment costs are a relevant concern given the competing demands on family and health care budgets. The authors evaluated the cost-effectiveness of three active treatments among adolescents with major depressive disorder. METHOD: Volunteers (N=439) ages 12 to 18 with a primary diagnosis of major depressive disorder participated in a randomized, controlled trial conducted at 13 U.S. academic and community clinics from 2000 to 2004. Subjects included those participants who did not drop out and had evaluable outcome and cost data at 12 weeks (N=369). Subjects were randomly assigned to 12 weeks of either fluoxetine alone (10-40 mg/day), CBT alone, CBT combined with fluoxetine (10-40 mg/day), or placebo (equivalent to 10-40 mg/day). Both placebo and fluoxetine were administered double-blind; CBT alone and CBT in combination with fluoxetine were administered unblinded. Societal cost per unit of improvement on the Children's Depression Rating Scale-Revised and cost per quality-adjusted life year (QALY) were compared. RESULTS: Results ranged from an incremental cost over placebo of $24,000 per QALY for treatment with fluoxetine to $123,000 per QALY for combination therapy treatment. The cost-effectiveness ratio for CBT treatment was not evaluable due to negative clinical effects. The models were robust on a variety of assumptions. CONCLUSIONS: Both fluoxetine and combination therapy are at least as cost-effective in the short-term as other treatments commonly used in primary care (using a threshold of $125,000/QALY). Fluoxetine is more cost effective than combination therapy after 12 weeks of treatment. PMID- 18413704 TI - Premorbid IQ in schizophrenia: a meta-analytic review. AB - OBJECTIVE: Over the past three decades, there have been significant changes in the diagnostic criteria for schizophrenia as well as changes in measurement of IQ. The last quantitative review of the literature on premorbid IQ in schizophrenia was published more than two decades ago. Since that time, there have been many published studies of data sets pertaining to this issue. The purpose of the present review was to provide an updated meta-analysis of premorbid IQ in individuals who later develop schizophrenia. METHOD: The authors performed a systematic literature search, which yielded 18 studies that met criteria for the meta-analysis. Inclusion criteria were 1) premorbid psychometric measures of IQ in subjects who were later diagnosed with schizophrenia, schizoaffective disorder, or schizophreniform disorder, 2) similar comparison data, and 3) sufficient data for calculation of an effect size. The analogue to the analysis of variance method was used to model between-study variance due to key study-design features. RESULTS: Overall, schizophrenia samples demonstrated a reliable, medium-sized impairment in premorbid IQ. The heterogeneity of effect sizes was minimal and almost exclusively the result of one study. Methodological differences, such as diagnostic criteria, type of IQ measure, sample ascertainment, and age at premorbid testing, contributed minimally to the effect size variance. A cross-sectional analysis of all studies by age and a descriptive review of studies that used repeated measures of IQ in a single sample did not support the presence of a relative decline in IQ during the premorbid period in individuals with schizophrenia. However, all studies with pre- and post-onset testing within the same sample suggested that a significant decline in the IQ of individuals with schizophrenia, relative to comparison subjects, was associated with the onset of frank psychosis. CONCLUSIONS: Years before the onset of psychotic symptoms, individuals with schizophrenia, as a group, demonstrate mean IQ scores approximately one-half of a standard deviation below that of healthy comparison subjects. PMID- 18413705 TI - A 6-week randomized, placebo-controlled trial of CP-316,311 (a selective CRH1 antagonist) in the treatment of major depression. AB - OBJECTIVE: The corticotropin-releasing hormone (CRH) system is implicated in the pathogenesis of several psychiatric disorders, including major depressive disorder. This study was designed to evaluate the safety and efficacy of CP 316,311, a selective nonpeptide antagonist of corticotropin-releasing hormone type 1 (CRH(1)) receptors, in the treatment of recurrent major depressive disorder. METHOD: Of a total of 167 patients with recurrent major depression who were screened, 123 were randomly assigned to receive 400 mg of CP-316,311 twice daily, or 100 mg of sertraline daily, or placebo in a 6-week fixed-dose, double blind, double-dummy, parallel-group, placebo- and sertraline-controlled trial. The primary efficacy analysis compared the change in score from baseline to endpoint on the 17-item Hamilton Depression Rating Scale (HAM-D) between the CP 316,311 and placebo groups. A group sequential design was used to support early trial termination based on efficacy or futility at a planned interim analysis. RESULTS: The evaluable data set for the interim analysis included 28 patients in the CP-316,311 group, 31 patients in the placebo group, and 30 patients in the sertraline group. In the interim analysis, the change from baseline in the HAM-D score at the final visit was not significantly different between the CP-316,311 and placebo groups, while change from baseline between the sertraline and placebo groups was significantly different. Given these results, futility was declared for CP-316,311 and the trial was terminated. CONCLUSIONS: Although CP-316,311 was safe and well tolerated in this study population, it failed to demonstrate efficacy in the treatment of major depression. PMID- 18413706 TI - Dissociated functional brain abnormalities of inhibition in boys with pure conduct disorder and in boys with pure attention deficit hyperactivity disorder. AB - OBJECTIVE: Inhibitory dysfunction may be a transdiagnostic etiopathophysiology of disruptive behavior disorders. Functional magnetic resonance imaging (fMRI) of inhibitory control has only been investigated in patients with attention deficit hyperactivity disorder (ADHD), including comorbidity with conduct disorder, showing frontal-striatal dysfunction. This study investigates differences and commonalities in functional neural networks mediating inhibitory control between medication-naive adolescents with pure conduct disorder and those with pure ADHD to identify biological markers that distinguish these clinically overlapping disorders. METHOD: Event-related fMRI was used to compare brain activation of 13 boys with noncomorbid conduct disorder, 20 with noncomorbid ADHD, and 20 normal boys during an individually adjusted tracking stop task that measures the neural substrates of inhibition and stopping failure. RESULTS: During successful inhibition, only patients with ADHD showed reduced activation in the left dorsolateral prefrontal cortex in relation to comparison subjects and patients with conduct disorder. During inhibition failures compared to go responses, both patient groups shared underactivation in the posterior cingulate gyrus in relation to comparison subjects. Patients with conduct disorder showed reduced activation in bilateral temporal-parietal regions compared to the other groups, which did not differ in this measure. CONCLUSIONS: Patients with pure ADHD or pure conduct disorder show qualitative differences in their brain abnormality patterns during inhibitory control. Inhibition-mediating prefrontal regions appear to be specifically reduced in ADHD, whereas posterior temporal-parietal, performance monitoring networks are specifically dysfunctional in conduct disorder. The findings provide pioneering evidence that distinct neurobiological abnormalities may be underlying the overlapping behavioral phenotype of the two disruptive disorders. PMID- 18413707 TI - Transference interpretations in dynamic psychotherapy: do they really yield sustained effects? AB - OBJECTIVE: Transference interpretation has remained a core ingredient in the psychodynamic tradition, despite limited empirical evidence for its effectiveness. In this study, the authors examined long-term effects of transference interpretations. METHOD: This was a randomized controlled clinical trial, dismantling design, plus follow-up evaluations 1 year and 3 years after treatment termination. One hundred outpatients seeking psychotherapy for depression, anxiety, personality disorders, and interpersonal problems were referred to the study therapists. Patients were randomly assigned to receive weekly sessions of dynamic psychotherapy for 1 year with or without transference interpretations. Five full sessions from each therapy were rated in order to document treatment fidelity. Outcome variables were the Psychodynamic Functioning Scales (clinician rated) and the Inventory of Interpersonal Problems (self report). Rating on the Quality of Object Relations Scale (lifelong pattern) and presence of a personality disorder were postulated moderators of treatment effects. Change over time was assessed using linear mixed models. RESULTS: Despite an absence of differential treatment efficacy, both treatments demonstrated significant improvement during treatment and also after treatment termination. However, patients with a lifelong pattern of poor object relations profited more from 1 year of therapy with transference interpretations than from therapy without transference interpretations. This effect was sustained throughout the 4-year study period. CONCLUSIONS: The goal of transference interpretation is sustained improvement of the patient's relationships outside of therapy. Transference interpretation seems to be especially important for patients with long-standing, more severe interpersonal problems. PMID- 18413708 TI - Semaphorin signaling in morphogenesis: found in translation. AB - Semaphorins play diverse roles in axon guidance and epithelial morphogenetic cell movements. In this issue of Genes & Development, Nukazuka and colleagues (1025 1036) show that semaphorins regulate Caenorhabditis elegans male tail morphogenesis by stimulating the translation of specific messages, including the actin-depolymerizing enzyme cofilin. PMID- 18413709 TI - Fine-tuning of hematopoietic stem cell homeostasis: novel role for ubiquitin ligase. AB - Homeostasis of hematopoietic stem cells (HSCs) is a tightly regulated process, controlled by intrinsic and extrinsic signals. Although a variety of molecules involved in HSC maintenance and self-renewal are known, it remains unclear how robust HSC homeostasis is achieved. In this issue of Genes & Development, Rathinam and colleagues (992-997) report a new player in HSC homeostasis, c-Cbl ubiquitin ligase. They show that this E3 ubiquitin ligase acts as a negative regulator of cytokine signaling. PMID- 18413710 TI - pRb and condensin--local control of global chromosome structure. AB - Rb mutants exhibit aneuploidy and aberrant chromosome structure during mitosis. In this issue of Genes & Development, a new paper from Longworth and colleagues (1011-1024) describes both physical and functional interactions between Drosophila Rbf1 and the dCAP-D3 subunit of condensin II. This work directly implicates the Rb family proteins in mitotic chromosome condensation and suggests that a failure in targeting condensin II to chromatin underlies the aneuploidy in rbf1 mutants. PMID- 18413711 TI - Small RNA guides for de novo DNA methylation in mammalian germ cells. AB - Germline genomic methylation is essential for gamete identity and integrity in mammals. The study by Kuramochi-Miyagawa and colleagues (908-917) in the previous issue of Genes & Development links the process of DNA methylation-dependent repression of retrotranspons with the presence of piwi-interacting RNAs (piRNAs) in fetal male germ cells undergoing de novo methylation. PMID- 18413712 TI - More than one way to skin . . . AB - Epithelial stem cells in the skin are specified during development and are governed by epithelial-mesenchymal interactions to differentially adopt the cell fates that enable them to form the epidermis, hair follicle, and sebaceous gland. In the adult, each of three epithelial lineages maintains their own stem cell population for self-renewal and normal tissue homeostasis. However, in response to injury, at least some of these stem cell niches can be mobilized to repair an epithelial tissue whose resident stem cells have been damaged. How do these stem cell populations respond to multiple signaling networks, activate migration, and proliferation, and differentiate along a specific lineage? Recent clues add new pieces to this multidimensional puzzle. Understanding how these stem cells maintain normal homeostasis and wound repair in the skin is particularly important, as these mechanisms, when defective, lead to skin tissue diseases including cancers. PMID- 18413713 TI - The E3 ubiquitin ligase c-Cbl restricts development and functions of hematopoietic stem cells. AB - Hematopoietic stem cells (HSCs) are multipotent progenitors that give rise to all types of blood cells. In the present study, we document that HSC development and functions are negatively regulated by the E3 ubiquitin ligase c-Cbl (casitas B cell lymphoma). HSCs of c-Cbl(-/-) mice exhibit augmented pool size, hyperproliferation, greater competence, and enhanced long-term repopulating capacity. Our mechanistic studies identified that c-Cbl(-/-) HSCs are hyperresponsive to thrombopoietin (TPO) and display elevated levels of STAT5 phosphorylation, thus leading to increased c-Myc expression. In essence, our data unequivocally identify c-Cbl as a novel negative regulator of developmental and functional properties of HSCs. PMID- 18413714 TI - Daxx represses RelB target promoters via DNA methyltransferase recruitment and DNA hypermethylation. AB - The apoptosis-modulating protein Daxx functions as a transcriptional repressor that binds to and suppresses the activity of nuclear factor-kappaB member RelB, among other transcription factors. The mechanism by which Daxx represses RelB target genes remains elusive. In this report, we demonstrate that Daxx controls epigenetic silencing of RelB target genes by DNA methylation. Daxx potently represses the RelB target genes dapk1, dapk3, c-flip, and birc3 (ciap2) at both the mRNA and protein levels. Recruitment of Daxx to target gene promoters, and its ability to repress them, is RelB-dependent, as shown by experiments using relB(-/-) cells. Importantly, methylation of target promoters is decreased in daxx(-/-) cells compared with daxx(+/+) cells, and stable transfection of daxx(-/ ) cells with Daxx restores DNA methylation. Furthermore, Daxx recruits DNA methyl transferase 1 (Dnmt1) to target promoters, resulting in synergistic repression. The observation that Daxx functions to target DNA methyltransferases onto RelB target sites in the genome provides a rare example of a gene-specific mechanism for epigenetic silencing. Given the documented role of several of the RelB regulated genes in diseases, particularly cancer, the findings have implications for developing therapeutic strategies based on epigenetic-modifying drugs. PMID- 18413715 TI - Semaphorin controls epidermal morphogenesis by stimulating mRNA translation via eIF2alpha in Caenorhabditis elegans. AB - Conserved semaphorin-plexin signaling systems govern various aspects of animal development, including axonal guidance in vertebrates and epidermal morphogenesis in Caenorhabditis elegans. Here we provide in vivo evidence that stimulation of mRNA translation via eukaryotic initiation factor 2alpha (eIF2alpha) is an essential downstream event of semaphorin signaling in C. elegans. In semaphorin/plexin mutants, a marked elevation in the phosphorylation of eIF2alpha is observed, which causes translation repression and is causally related to the morphological epidermal phenotype in the mutants. Conversely, removal of constraints on translation by genetically reducing the eIF2alpha phosphorylation largely bypasses requirement for the semaphorin signal in epidermal morphogenesis. We also identify an actin-depolymerizing factor/cofilin, whose expression in the mutants is predominantly repressed, as a major translational target of semaphorin signaling. Thus, our results reveal a physiological significance for translation of mRNAs for cytoskeletal regulators, linking environmental cues to cytoskeletal rearrangement during cellular morphogenesis in vivo. PMID- 18413716 TI - Translation of ASH1 mRNA is repressed by Puf6p-Fun12p/eIF5B interaction and released by CK2 phosphorylation. AB - Translational repression during mRNA transport is essential for spatial restriction of protein production. In the yeast Saccharomyces cerevisae, silencing of ASH1 mRNA before it is localized to the bud cortex in late anaphase is critical for asymmetric segregation of Ash1p to the daughter cell nucleus. Puf6p, an ASH1 mRNA-binding protein, has been implicated in this process as a translational repressor, but the underlying mechanism is unknown. Here, we used yeast extract-based in vitro translation assays, which recapitulate translation and phosphorylation, to characterize the mechanism of Puf6p-mediated translational regulation. We report that Puf6p interferes with the conversion of the 48S complex to the 80S complex during initiation, and this repression by Puf6p is mediated through the general translation factor eIF5B (Fun12p in S. cerevisiae). Puf6p interacts with Fun12p via the PUF domain, and this interaction is RNA-dependent and essential for translational repression by Puf6p. This repression is relieved by phosphorylation of the N-terminal region of Puf6p mediated by protein kinase CK2 (casein kinase II). Inhibition of phosphorylation at Ser31, Ser34, and Ser35 of Puf6p increases its translational repression and results in ASH1 mRNA delocalization. Our results indicate that Puf6p suppresses the translation initiation of ASH1 mRNA via interaction with Fun12p during its transport, and this repression can be released by CK2 phosphorylation in the N terminal region of Puf6p when the mRNA reaches the bud tip. PMID- 18413717 TI - Efficient termination of transcription by RNA polymerase I requires the 5' exonuclease Rat1 in yeast. AB - During transcription termination by RNA polymerase II on protein-coding genes, the nuclear 5' exonuclease Rat1/Xrn2 degrades the nascent transcript downstream from the polyadenylation site and "torpedoes" the polymerase. We report that the activity of Rat1 is also required for efficient termination by RNA polymerase I (Pol I) on the rDNA. In strains lacking catalytically active Rat1 or its cofactor Rai1, Pol I reads through the major, "Reb1-dependent" terminator (T1) but stops downstream at the "fail-safe" terminator (T2) and replication fork barrier (RFB). The absence of both Rat1 and the RFB-binding protein Fob1 increased Pol I read through of T2 and the RFB. We propose that cotranscriptional cleavage of the pre rRNA by the endonuclease Rnt1 generates a loading site for the Rat1/Rai1 complex, which then degrades the nascent transcript. When Rat1 catches Pol I, which is predicted to be paused at T1, transcription is terminated. PMID- 18413718 TI - Budding yeast RNA polymerases I and II employ parallel mechanisms of transcriptional termination. AB - Both RNA polymerase I and II (Pol I and Pol II) in budding yeast employ a functionally homologous "torpedo-like" mechanism to promote transcriptional termination. For two well-defined Pol II-transcribed genes, CYC1 and PMA1, we demonstrate that both Rat1p exonuclease and Sen1p helicase are required for efficient termination by promoting degradation of the nascent transcript associated with Pol II, following mRNA 3' end processing. Similarly, Pol I termination relies on prior Rnt1p cleavage at the 3' end of the pre-rRNA 35S transcript. This is followed by the combined actions of Rat1p and Sen1p to degrade the Pol I-associated nascent transcript that consequently promote termination in the downstream rDNA spacer sequence. Our data suggest that the previously defined in vitro Pol I termination mechanism involving the action of the Reb1p DNA-binding factor to "road-block" Pol I transcription close to the termination region may have overlooked more complex in vivo molecular processes. PMID- 18413719 TI - Crystal structure of the Mus81-Eme1 complex. AB - The Mus81-Eme1 complex is a structure-specific endonuclease that plays an important role in rescuing stalled replication forks and resolving the meiotic recombination intermediates in eukaryotes. We have determined the crystal structure of the Mus81-Eme1 complex. Both Mus81 and Eme1 consist of a central nuclease domain, two repeats of the helix-hairpin-helix (HhH) motif at their C terminal region, and a linker helix. While each domain structure resembles archaeal XPF homologs, the overall structure is significantly different from those due to the structure of a linker helix. We show that a flexible intradomain linker that formed with 36 residues in the nuclease domain of Eme1 is essential for the recognition of DNA. We identified several basic residues lining the outer surface of the active site cleft of Mus81 that are involved in the interaction with a flexible arm of a nicked Holliday junction (HJ). These interactions might contribute to the optimal positioning of the opposite junction across the nick into the catalytic site, which provided the basis for the "nick and counternick" mechanism of Mus81-Eme1 and for the nicked HJ to be the favored in vitro substrate of this enzyme. PMID- 18413720 TI - The quest for the 1p36 tumor suppressor. AB - Genomic analyses of late-stage human cancers have uncovered deletions encompassing 1p36, thereby providing an extensive body of literature supporting the idea that a potent tumor suppressor resides in this interval. Although several genes have been proposed as 1p36 candidate tumor suppressors, convincing evidence that their encoded products protect from cancer has been scanty. A recent functional study identified chromodomain helicase DNA-binding protein 5 (CHD5) as a novel tumor suppressor mapping to 1p36. Here, we discuss evidence supporting the tumor-suppressive role of CHD5. Together, these findings suggest that strategies designed to enhance CHD5 activity could provide novel approaches for treating a broad range of human malignancies. PMID- 18413721 TI - Role of the aggresome pathway in cancer: targeting histone deacetylase 6 dependent protein degradation. AB - Misfolded or aggregated proteins have two fates: they are either refolded with the help of chaperones or degraded by the proteasome. Cells also have an alternative pathway that involves intracellular "storage bins" for misfolded intracellular proteins known as aggresomes. Aggresomes recruit motor proteins that transport misfolded or aggregated proteins to chaperones and proteasomes for subsequent destruction. There is emerging evidence that inhibiting the aggresome pathway leads to accumulation of misfolded proteins and apoptosis in tumor cells through autophagy. We discuss the role of aggresomes in cancer and the potential to target this pathway for therapy. PMID- 18413722 TI - Tumor escape mechanism governed by myeloid-derived suppressor cells. AB - T-cell nonresponsiveness is a critical factor in immune escape and myeloid derived suppressor cells play a major role in organizing this phenomenon. Recent findings indicate that myeloid-derived suppressor cells can induce antigen specific CD8(+) T-cell tolerance through a posttranslation mechanism which involves modification (nitration) of CD8 and the T-cell receptor itself on the T cell surface. Elucidation of this mechanism of T-cell tolerance offers new opportunities for therapeutic corrections of immune escape in cancer. PMID- 18413723 TI - A requirement for DICER to maintain full promoter CpG island hypermethylation in human cancer cells. AB - Promoter hypermethylation is a prevalent phenomenon, found in virtually all cancer types studied thus far, and accounts for tumor suppressor gene silencing in the absence of genetic mutations. The mechanism behind the establishment and maintenance of such aberrant hypermethylation has been under intense study. Here, we have uncovered a link between aberrant gene silencing associated with promoter CpG island DNA methylation and the siRNA/miRNA processing enzyme, DICER, in human cancer cells. By comparing demethylated HCT116 colon cancer cells with HCT116 cells genetically rendered hypomorphic for DICER, we identified a group of epigenetically silenced genes that became reactivated in the absence of functional DICER. This reactivation is associated with a dramatic loss of localized promoter DNA hypermethylation. Thus, intact DICER is required to maintain full promoter DNA hypermethylation of select epigenetically silenced loci in human cancer cells. PMID- 18413724 TI - BCR/ABL inhibits mismatch repair to protect from apoptosis and induce point mutations. AB - BCR/ABL kinase-positive chronic myelogenous leukemia (CML) cells display genomic instability leading to point mutations in various genes including bcr/abl and p53, eventually causing resistance to imatinib and malignant progression of the disease. Mismatch repair (MMR) is responsible for detecting misincorporated nucleotides, resulting in excision repair before point mutations occur and/or induction of apoptosis to avoid propagation of cells carrying excessive DNA lesions. To assess MMR activity in CML, we used an in vivo assay using the plasmid substrate containing enhanced green fluorescent protein (EGFP) gene corrupted by T:G mismatch in the start codon; therefore, MMR restores EGFP expression. The efficacy of MMR was reduced approximately 2-fold in BCR/ABL positive cell lines and CD34(+) CML cells compared with normal counterparts. MMR was also challenged by N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), which generates O(6)-methylguanine and O(4)-methylthymine recognized by MMR system. Impaired MMR activity in leukemia cells was associated with better survival, accumulation of p53 but not of p73, and lack of activation of caspase 3 after MNNG treatment. In contrast, parental cells displayed accumulation of p53, p73, and activation of caspase 3, resulting in cell death. Ouabain-resistance test detecting mutations in the Na(+)/K(+) ATPase was used to investigate the effect of BCR/ABL kinase-mediated inhibition of MMR on mutagenesis. BCR/ABL-positive cells surviving the treatment with MNNG displayed approximately 15-fold higher mutation frequency than parental counterparts and predominantly G:C-->A:T and A:T ->G:C mutator phenotype typical for MNNG-induced unrepaired lesions. In conclusion, these results suggest that BCR/ABL kinase abrogates MMR activity to inhibit apoptosis and induce mutator phenotype. PMID- 18413725 TI - Secondary BRCA1 mutations in BRCA1-mutated ovarian carcinomas with platinum resistance. AB - Although ovarian carcinomas with mutated BRCA1 or BRCA2 are sensitive to platinum compounds, such carcinomas eventually develop platinum resistance. Previously, we showed that acquired resistance to cisplatin in BRCA2-mutated tumors can be mediated by secondary intragenic mutations in BRCA2 that restore the wild-type BRCA2 reading frame. Here, we show that secondary mutations of BRCA1 also occur in BRCA1-mutated ovarian cancer with platinum resistance. We evaluated nine recurrent BRCA1-mutated ovarian cancers previously treated with platinum compounds, including five with acquired platinum resistance, one with primary platinum resistance, and three with platinum sensitivity. Four of the six recurrent platinum-resistant tumors had developed secondary genetic changes in BRCA1 that restored the reading frame of the BRCA1 protein, whereas none of the three platinum-sensitive recurrent tumors developed BRCA1 sequence alterations. We immunohistochemically confirmed restored expression of BRCA1 protein in two cases with secondary mutations. Intriguingly, the case with primary platinum resistance showed back mutation of BRCA1 in the primary tumor and showed another secondary mutation in the recurrent tumor. Our results suggest that secondary mutations in BRCA1 can mediate resistance to platinum in BRCA1-mutated ovarian tumors. PMID- 18413726 TI - Identification of let-7-regulated oncofetal genes. AB - MicroRNAs (miRNA) are small RNA molecules of approximately 20 to 22 nucleotides that reduce expression of proteins through mRNA degradation and/or translational silencing. Each known miRNA has a large number of predicted targets. Members of the let-7/miR-98 family of miRNAs are up-regulated at the end of embryonic development. Let-7 is often down-regulated early during cancer development, suggesting that let-7-regulated oncofetal genes (LOG) may become reexpressed in cancer cells. Using comparative bioinformatics, we have identified 12 conserved LOGs that include HMGA2 and IMP-1/CRD-BP. IMP-1 has growth-promoting activities through stabilization of c-myc mRNA. We experimentally confirmed that IMP-1 is a direct let-7 target that promotes cell growth and motility of tumor cells, and we confirmed by proteomics analysis that IMP-1 and HMGA2 are major miRNA targets. Our data suggest that a substantial part of the growth inhibitory activities of let-7 comes from suppressing the expression of IMP-1. LOGs could be novel therapeutic targets and potential biomarkers for cancer treatment. PMID- 18413727 TI - A role for TAZ in migration, invasion, and tumorigenesis of breast cancer cells. AB - TAZ (WWTR1), identified as a 14-3-3 binding protein with a PDZ binding motif, modulates mesenchymal stem cell differentiation. We now show that TAZ plays a critical role in the migration, invasion, and tumorigenesis of breast cancer cells. TAZ is conspicuously expressed in human breast cancer cell lines in which its expression levels generally correlate with the invasiveness of cancer cells. Overexpression of TAZ in low-expressing MCF10A cells causes morphologic changes characteristic of cell transformation and promotes cell migration and invasion. Conversely, RNA interference-mediated knockdown of TAZ expression in MCF7 and Hs578T cells reduces cell migration and invasion. TAZ knockdown in MCF7 cells also retards anchorage-independent growth in soft agar and tumorigenesis in nude mice. Significantly, TAZ is overexpressed in approximately 20% of breast cancer samples. These results indicate that TAZ plays a role in the migration, invasion, and tumorigenesis of breast cancer cells and thus presents a novel target for the detection and treatment of breast cancer. PMID- 18413728 TI - Cyclin D1 and CDK4 activity contribute to the undifferentiated phenotype in neuroblastoma. AB - Genomic aberrations of Cyclin D1 (CCND1), CDK4, and CDK6 in neuroblastoma indicate that dysregulation of the G(1) entry checkpoint is an important cell cycle aberration in this pediatric tumor. Here, we report that analysis of Affymetrix expression data of primary neuroblastic tumors shows an extensive overexpression of Cyclin D1, which correlates with histologic subgroups. Immunohistochemical analysis showed overexpression of Cyclin D1 in neuroblasts and low Cyclin D1 expression in all cell types in ganglioneuroma. This suggests an involvement of G(1)-regulating genes in neuronal differentiation processes which we further evaluated using RNA interference against Cyclin D1 and its kinase partners CDK4 and CDK6 in several neuroblastoma cell lines. The Cyclin D1 and CDK4 knockdown resulted in pRb pathway inhibition as shown by an almost complete disappearance of CDK4/CDK6-specific pRb phosphorylation, reduction of E2F transcriptional activity, and a decrease of Cyclin A protein levels. Phenotype analysis showed a significant reduction in cell proliferation, a G(1) specific cell cycle arrest, and, moreover, an extensive neuronal differentiation. Affymetrix microarray profiling of small interfering RNA-treated cells revealed a shift in expression profile toward a neuronal phenotype. Several new potential downstream players are identified. We conclude that neuroblastoma functionally depend on overexpression of G(1)-regulating genes to maintain their undifferentiated phenotype. PMID- 18413729 TI - p53 Loss synergizes with estrogen and papillomaviral oncogenes to induce cervical and breast cancers. AB - Whereas the tumor suppressor p53 gene is frequently mutated in most human cancers, this is not the case in human papillomavirus (HPV)-associated cancers, presumably because the viral E6 oncoprotein inactivates the p53 protein. The ability of E6 to transform cells in tissue culture and induce cancers in mice correlates in part with its ability to inactivate p53. In this study, we compared the expression of the HPV16 E6 oncogene to the conditional genetic disruption of p53 in the context of a mouse model for cervical cancer in which estrogen is a critical cofactor. Nearly all of the K14Crep53(f/f) mice treated with estrogen developed cervical cancer, a stark contrast to its complete absence in like treated K14E6(WT)p53(f/f) mice, indicating that HPV16 E6 must only partially inactivate p53. p53-independent activities of E6 also contributed to carcinogenesis, but in the female reproductive tract, these activities were manifested only in the presence of the HPV16 E7 oncogene. Interestingly, treatment of K14Crep53(f/f) mice with estrogen also resulted in mammary tumors after only a short latency, many of which were positive for estrogen receptor alpha. The majority of these mammary tumors were of mixed cell types, suggestive of their originating from a multipotent progenitor. Furthermore, a subset of mammary tumors arising in the estrogen-treated, p53-deficient mammary glands exhibited evidence of an epithelial to mesenchymal transition. These data show the importance of the synergy between estrogen and p53 insufficiency in determining basic properties of carcinogenesis in hormone-responsive tissues, such as the breast and the reproductive tract. PMID- 18413730 TI - Bile acid exposure up-regulates tuberous sclerosis complex 1/mammalian target of rapamycin pathway in Barrett's-associated esophageal adenocarcinoma. AB - Barrett's esophagus, a columnar metaplasia of the lower esophagus epithelium related to gastroesophageal reflux disease, is the strongest known risk factor for the development of esophageal adenocarcinoma (EAC). Understanding the signal transduction events involved in esophageal epithelium carcinogenesis may provide insights into the origins of EAC and may suggest new therapies. To elucidate the molecular pathways of bile acid-induced tumorigenesis, the newly identified inflammation-associated signaling pathway involving I kappaB kinases beta (IKK beta), tuberous sclerosis complex 1 (TSC1), and mammalian target of rapamycin (mTOR) downstream effector S6 kinase (S6K1) was confirmed to be activated in immortalized Barrett's CPC-A and CPC-C cells and esophageal cancer SEG-1 and BE3 cells. Phosphorylation of TSC1 and S6K1 was induced in response to bile acid stimulation. Treatment of these cells with the mTOR inhibitor rapamycin or the IKK beta inhibitor Bay 11-7082 suppressed bile acid-induced cell proliferation and anchorage-independent growth. We next used an orthotopic rat model to evaluate the role of bile acid in the progression of Barrett's esophagus to EAC. Of interest, we found high expression of phosphorylated IKK beta (pIKK beta) and phosphorylated S6K1 (pS6K1) in tumor tissues and the Barrett's epithelium compared with normal epithelium. Furthermore, immunostaining of clinical EAC tissue specimens revealed that pIKK beta expression was strongly correlated with pS6K1 level. Together, these results show that bile acid can deregulate TSC1/mTOR through IKK beta signaling, which may play a critical role in EAC progression. In addition, Bay 11-7082 and rapamycin may potentially be chemopreventive drugs against Barrett's esophagus-associated EAC. PMID- 18413731 TI - Analysis of the mechanisms mediating tumor-specific changes in gene expression in human liver tumors. AB - There is widespread interest in efficient characterization of differences between tumor and normal samples. Here, we show an effective methodology for genome-scale characterization of tumors. Using matched normal and tumor samples from liver cancer patients, as well as non-cancer-related normal liver tissue, we first determined changes in gene expression as monitored on RNA expression arrays. We identified several hundred mRNAs that were consistently changed in the tumor samples. To characterize the mechanisms responsible for creation of the tumor specific transcriptome, we performed chromatin immunoprecipitation on microarray experiments to assay binding of RNA polymerase II, H3me3K27, and H3me3K9 and DNA methylation in 25,000 promoter regions. These experiments identified changes in active and silenced regions of the genome in the tumor cells. Finally, we used a "virtual comparative genomic hybridization" method to identify copy number alterations in the tumor samples. Through comparison of RNA polymerase II binding, chromatin structure, DNA methylation, and copy number changes, we suggest that the major contributor to creation of the liver tumor transcriptome was changes in gene copy number. PMID- 18413732 TI - Broad overexpression of ribonucleotide reductase genes in mice specifically induces lung neoplasms. AB - Ribonucleotide reductase (RNR) catalyzes the rate-limiting step in nucleotide biosynthesis and plays a central role in genome maintenance. Although a number of regulatory mechanisms govern RNR activity, the physiologic effect of RNR deregulation had not previously been examined in an animal model. We show here that overexpression of the small RNR subunit potently and selectively induces lung neoplasms in transgenic mice and is mutagenic in cultured cells. Combining RNR deregulation with defects in DNA mismatch repair, the cellular mutation correction system, synergistically increased RNR-induced mutagenesis and carcinogenesis. Moreover, the proto-oncogene K-ras was identified as a frequent mutational target in RNR-induced lung neoplasms. Together, these results show that RNR deregulation promotes lung carcinogenesis through a mutagenic mechanism and establish a new oncogenic activity for a key regulator of nucleotide metabolism. Importantly, RNR-induced lung neoplasms histopathologically resemble human papillary adenocarcinomas and arise stochastically via a mutagenic mechanism, making RNR transgenic mice a valuable model for lung cancer. PMID- 18413733 TI - Genome-wide promoter analysis uncovers portions of the cancer methylome. AB - DNA methylation has a role in mediating epigenetic silencing of CpG island genes in cancer and other diseases. Identification of all gene promoters methylated in cancer cells "the cancer methylome" would greatly advance our understanding of gene regulatory networks in tumorigenesis. We previously described a new method of identifying methylated tumor suppressor genes based on pharmacologic unmasking of the promoter region and detection of re-expression on microarray analysis. In this study, we modified and greatly improved the selection of candidates based on new promoter structure algorithm and microarray data generated from 20 cancer cell lines of 5 major cancer types. We identified a set of 200 candidate genes that cluster throughout the genome of which 25 were previously reported as harboring cancer-specific promoter methylation. The remaining 175 genes were tested for promoter methylation by bisulfite sequencing or methylation-specific PCR (MSP). Eighty-two of 175 (47%) genes were found to be methylated in cell lines, and 53 of these 82 genes (65%) were methylated in primary tumor tissues. From these 53 genes, cancer-specific methylation was identified in 28 genes (28 of 53; 53%). Furthermore, we tested 8 of the 28 newly identified cancer-specific methylated genes with quantitative MSP in a panel of 300 primary tumors representing 13 types of cancer. We found cancer-specific methylation of at least one gene with high frequency in all cancer types. Identification of a large number of genes with cancer-specific methylation provides new targets for diagnostic and therapeutic intervention, and opens fertile avenues for basic research in tumor biology. PMID- 18413734 TI - Targeted deletion of Prkar1a reveals a role for protein kinase A in mesenchymal to-epithelial transition. AB - Dysregulation of protein kinase A (PKA) activity, caused by loss of function mutations in PRKAR1A, is known to induce tumor formation in the inherited tumor syndrome Carney complex (CNC) and is also associated with sporadic tumors of the thyroid and adrenal. We have previously shown that Prkar1a(+/-) mice develop schwannomas reminiscent of those seen in CNC and that similar tumors are observed in tissue-specific knockouts (KO) of Prkar1a targeted to the neural crest. Within these tumors, we have previously described the presence of epithelial islands, although the nature of these structures was unclear. In this article, we report that these epithelial structures are derived from KO cells originating in the neural crest. Analysis of the mesenchymal marker vimentin revealed that this protein was markedly down-regulated not only from the epithelial islands, but also from the tumor as a whole, consistent with mesenchymal-to-epithelial transition (MET). In vitro, Prkar1a null primary mouse embryonic fibroblasts, which display constitutive PKA signaling, also showed evidence for MET, with a loss of vimentin and up-regulation of the epithelial marker E-cadherin. Reduction of vimentin protein occurred at the posttranslational level and was rescued by proteasomal inhibition. Finally, this down-regulation of vimentin was recapitulated in the adrenal nodules of CNC patients, confirming an unexpected and previously unrecognized role for PKA in MET. PMID- 18413735 TI - Androgen receptor overexpression in prostate cancer linked to Pur alpha loss from a novel repressor complex. AB - Increased androgen receptor (AR) expression and activity are pivotal for androgen independent (AI) prostate cancer (PC) progression and resistance to androgen deprivation therapy. We show that a novel transcriptional repressor complex that binds a specific sequence (repressor element) in the AR gene 5'-untranslated region contains Pur alpha and hnRNP-K. Pur alpha expression, its nuclear localization, and its AR promoter association, as determined by chromatin immunoprecipitation analysis, were found to be significantly diminished in AI LNCaP cells and in hormone-refractory human PCs. Transfection of AI cells with a plasmid that restored Pur alpha expression reduced AR at the transcription and protein levels. Pur alpha knockdown in androgen-dependent cells yielded higher AR and reduced p21, a gene previously shown to be under negative control of AR. These changes were linked to increased proliferation in androgen-depleted conditions. Treatment of AI cells with histone deacetylase and DNA methylation inhibitors restored Pur alpha protein and binding to the AR repressor element. This correlated with decreased AR mRNA and protein levels and inhibition of cell growth. Pur alpha is therefore a key repressor of AR transcription and its loss from the transcriptional repressor complex is a determinant of AR overexpression and AI progression of PC. The success in restoring Pur alpha and the repressor complex function by pharmacologic intervention opens a promising new therapeutic approach for advanced PC. PMID- 18413736 TI - Novel and highly recurrent chromosomal alterations in Sezary syndrome. AB - This study was designed to identify highly recurrent genetic alterations typical of Sezary syndrome (Sz), an aggressive cutaneous T-cell lymphoma/leukemia, possibly revealing pathogenetic mechanisms and novel therapeutic targets. High resolution array-based comparative genomic hybridization was done on malignant T cells from 20 patients. Expression levels of selected biologically relevant genes residing within loci with frequent copy number alteration were measured using quantitative PCR. Combined binary ratio labeling-fluorescence in situ hybridization karyotyping was done on malignant cells from five patients. Minimal common regions with copy number alteration occurring in at least 35% of patients harbored 15 bona fide oncogenes and 3 tumor suppressor genes. Based on the function of the identified oncogenes and tumor suppressor genes, at least three molecular mechanisms are relevant in the pathogenesis of Sz. First, gain of cMYC and loss of cMYC antagonists (MXI1 and MNT) were observed in 75% and 40% to 55% of patients, respectively, which were frequently associated with deregulated gene expression. The presence of cMYC/MAX protein heterodimers in Sezary cells was confirmed using a proximity ligation assay. Second, a region containing TP53 and genome maintenance genes (RPA1/HIC1) was lost in the majority of patients. Third, the interleukin 2 (IL-2) pathway was affected by gain of STAT3/STAT5 and IL-2 (receptor) genes in 75% and 30%, respectively, and loss of TCF8 and DUSP5 in at least 45% of patients. In sum, the Sz genome is characterized by gross chromosomal instability with highly recurrent gains and losses. Prominent among deregulated genes are those encoding cMYC, cMYC-regulating proteins, mediators of MYC-induced apoptosis, and IL-2 signaling pathway components. PMID- 18413737 TI - Distinct RAD51 associations with RAD52 and BCCIP in response to DNA damage and replication stress. AB - RAD51 has critical roles in homologous recombination (HR) repair of DNA double strand breaks (DSB) and restarting stalled or collapsed replication forks. In yeast, Rad51 function is facilitated by Rad52 and other "mediators." Mammalian cells express RAD52, but BRCA2 may have supplanted RAD52 in mediating RAD51 loading onto ssDNA. BCCIP interacts with BRCA2, and both proteins are important for RAD51 focus formation after ionizing radiation and HR repair of DSBs. Nonetheless, mammalian RAD52 shares biochemical activities with yeast Rad52, including RAD51 binding and single-strand annealing, suggesting a conserved role in HR. Because RAD52 and RAD51 associate, and RAD51 and BCCIP associate, we investigated the colocalization of RAD51 with BCCIP and RAD52 in human cells. We found that RAD51 colocalizes with BCCIP early after ionizing radiation, with RAD52 later, and there was little colocalization of BCCIP and RAD52. RAD52 foci are induced to a greater extent by hydroxyurea, which stalls replication forks, than by ionizing radiation. Using fluorescence recovery after photo bleaching, we show that RAD52 mobility is reduced to a greater extent by hydroxyurea than ionizing radiation. However, BCCIP showed no changes in mobility after hydroxyurea or ionizing radiation. We propose that BCCIP-dependent repair of DSBs by HR is an early RAD51 response to ionizing radiation-induced DNA damage, and that RAD52-dependent HR occurs later to restart a subset of blocked or collapsed replication forks. RAD52 and BRCA2 seem to act in parallel pathways, suggesting that targeting RAD52 in BRCA2-deficient tumors may be effective in treating these tumors. PMID- 18413738 TI - MUC1 expression is regulated by DNA methylation and histone H3 lysine 9 modification in cancer cells. AB - MUC1 is a transmembrane mucin that is highly expressed in various cancers and correlates with malignant potential. Important cancer-related genes such as p16 and E-cadherin are controlled epigenetically; however, MUC1 has been overlooked in epigenetics. Herein, we provide the first report that MUC1 gene expression is regulated by DNA methylation and histone H3 lysine 9 (H3-K9) modification of the MUC1 promoter. The recently developed MassARRAY assay was performed to investigate the DNA methylation status of 184 CpG sites from -2,753 to +263. Near the transcriptional start site, the DNA methylation level of MUC1-negative cancer cell lines (e.g., MDA-MB-453) was high, whereas that of MUC1-positive cell lines (e.g., MCF-7) was low. Histone H3-K9 modification status was also closely related to MUC1 gene expression. Furthermore, MUC1 mRNA expression in MUC1-negative cells was restored by treatment with the DNA methylation inhibitor 5-aza-2' deoxycytidine. Our results indicate that DNA methylation and histone H3-K9 modification in the 5' flanking region play a critical role in MUC1 gene expression, and this study defines MUC1 as a new member of the class of epigenetically controlled genes. An understanding of the epigenetic changes of MUC1 may be of importance for diagnosis of carcinogenic risk and prediction of outcome for cancer patients. PMID- 18413739 TI - Breast carcinoma-associated fibroblasts and their counterparts display neoplastic specific changes. AB - It has become clear that the initiation and progression of carcinomas depend not only on alterations in epithelial cells, but also on changes in their microenvironment. To identify these changes, we have undertaken cellular and molecular characterization of carcinoma-associated fibroblasts (CAF) and their tumor counterpart fibroblasts (TCF) isolated from 12 breast cancer patients. Normal breast fibroblasts (NBF) from plastic surgery were used as normal control. We present evidence that both CAFs and TCFs are myofibroblasts and show tumor associated features. Indeed, the p53/p21 response pathway to gamma-rays was defective in 70% CAFs, whereas it was normal in all the TCF and NBF cells. In addition, the basal levels of the p53 and p21 proteins were significantly low in 83% of CAFs and modulated in the majority of TCFs compared with NBFs. Interestingly, both TCFs and CAFs expressed high levels of the cancer marker survivin and consequently exhibited high resistance to cisplatin and UV light. Moreover, most CAFs were positive for the proliferation marker Ki-67 and exhibited high proliferation rate compared with NBFs and TCFs. However, proliferating cell nuclear antigen was highly expressed in both CAFs and TCFs. Using the two-dimensional gel electrophoresis technique, we have also shown that CAF, TCF, and NBF cells present different proteome profiles, with many proteins differentially expressed between these cells. Taken together these results indicate that different genetic alterations can occur in breast CAFs and their corresponding adjacent counterparts, showing the important role that stroma could play in breast carcinogenesis and treatment. PMID- 18413740 TI - DNA methyltransferase 1 and 3B activate BAG-1 expression via recruitment of CTCFL/BORIS and modulation of promoter histone methylation. AB - In a previous genomic analysis, using somatic methyltransferase (DNMT) knockout cells, we showed that hypomethylation decreased the expression of as many genes as were observed to increase, suggesting a previously unknown mechanism for epigenetic regulation. To address this idea, the expression of the BAG family genes was used as a model. These genes were used because their expression was decreased in DNMT1(-/-), DNMT3B(-/-), and double knockout cells and increased in DNMT1-overexpressing and DNMT3B-overexpressing cells. Chromatin immunoprecipitation analysis of the BAG-1 promoter in DNMT1-overexpressing or DNMT3B-overexpressing cells showed a permissive dimethyl-H3-K4/dimethyl-H3-K9 chromatin status associated with DNA-binding of CTCFL/BORIS, as well as increased BAG-1 expression. In contrast, a nonpermissive dimethyl-H3-K4/dimethyl-H3-K9 chromatin status was associated with CTCF DNA-binding and decreased BAG-1 expression in the single and double DNMT knockout cells. BORIS short hairpin RNA knockdown decreased both promoter DNA-binding, as well as BAG-1 expression, and changed the dimethyl-H3-K4/dimethyl-H3-K9 ratio to that characteristic of a nonpermissive chromatin state. These results suggest that DNMT1 and DNMT3B regulate BAG-1 expression via insulator protein DNA-binding and chromatin dynamics by regulating histone dimethylation. PMID- 18413741 TI - Genistein induces the p21WAF1/CIP1 and p16INK4a tumor suppressor genes in prostate cancer cells by epigenetic mechanisms involving active chromatin modification. AB - Genistein (4',5,7-trihydroxyisoflavone) is the most abundant isoflavone found in the soybean. The effects of genistein on various cancer cell lines have been extensively studied but the precise molecular mechanisms are not known. We report here the epigenetic mechanism of the action of genistein on androgen-sensitive (LNCaP) and androgen-insensitive (DuPro) human prostate cancer cell lines. Genistein induced the expression of tumor suppressor genes p21 (WAF1/CIP1/KIP1) and p16 (INK4a) with a concomitant decrease in cyclins. There was a G(0)-G(1) cell cycle arrest in LNCaP cells and a G(2)-M arrest in DuPro cells after genistein treatment. Genistein also induced apoptosis in DuPro cells. DNA methylation analysis revealed the absence of p21 promoter methylation in both cell lines. The effect of genistein on chromatin remodeling has not been previously reported. We found that genistein increased acetylated histones 3, 4, and H3/K4 at the p21 and p16 transcription start sites. Furthermore, we found that genistein treatment also increased the expression of histone acetyl transferases that function in transcriptional activation. This is the first report on epigenetic regulation of various genes by genistein through chromatin remodeling in prostate cancer. Altogether, our data provide new insights into the epigenetic mechanism of the action of genistein that may contribute to the chemopreventive activity of this dietary isoflavone and have important implications for epigenetic therapy. PMID- 18413742 TI - Matrix metalloproteinase-8 functions as a metastasis suppressor through modulation of tumor cell adhesion and invasion. AB - Collagenase-2 (matrix metalloproteinase-8, MMP-8) is an MMP mainly produced by neutrophils and associated with many inflammatory conditions. We have previously described that MMP-8 plays a protective role in cancer through its ability to regulate the inflammatory response induced by carcinogens. Moreover, it has been reported that experimental manipulation of the expression levels of this enzyme alters the metastatic behavior of human breast cancer cells. In this work, we have used mutant mice deficient in MMP-8 and syngenic melanoma and lung carcinoma tumor cells lines overexpressing this enzyme to further explore the putative antimetastatic potential of MMP-8. We report herein that MMP-8 prevents metastasis formation through the modulation of tumor cell adhesion and invasion. Thus, tumor cells overexpressing MMP-8 have an increased adhesion to extracellular matrix proteins, whereas their invasive ability through Matrigel is substantially reduced when compared with control cells. Analysis of MMP-8 in breast cancer patients revealed that the expression of this metalloproteinase by breast tumors correlates with a lower incidence of lymph node metastasis and confers good prognosis to these patients. On this basis, we propose that MMP-8 is a tumor protective factor, which also has the ability to reduce the metastatic potential of malignant cells in both mice and human. PMID- 18413743 TI - Epigenetic inactivation of the canonical Wnt antagonist SRY-box containing gene 17 in colorectal cancer. AB - SRY-box containing gene 17 (Sox17) is a member of the high mobility group (HMG) transcription factor superfamily, which plays critical roles in the regulation of development and stem/precursor cell function, at least partly through repression of Wnt pathway activity. Modulators controlling aberrant Wnt signaling activation are frequently disrupted in human cancers through complementary effects of epigenetic and genetic changes. Our recent global analysis of CpG island hypermethylation and gene expression in colorectal cancer (CRC) cell lines revealed that SOX17 gene silencing is associated with DNA hypermethylation of a CpG island in the promoter region. Here, we report that CpG island methylation dependent silencing of SOX17 occurs in 100% of CRC cell lines, 86% of colorectal adenomas, 100% of stage I and II CRC, 89% of stage III CRC, 89% of primary esophageal cancer, and 50% of non-small cell lung cancer. Overexpression of SOX17 in HCT116 CRC cells inhibits colony growth and beta-catenin/T-cell factor dependent transcription. Structure-based deletion analysis further shows the presence of a Wnt signaling repression domain in the SOX17 HMG box. Together, our studies suggest that SOX17 is a negative modulator of canonical Wnt signaling, and that SOX17 silencing due to promoter hypermethylation is an early event during tumorigenesis and may contribute to aberrant activation of Wnt signaling in CRC. PMID- 18413744 TI - MicroRNAs 221 and 222 bypass quiescence and compromise cell survival. AB - MicroRNAs (miRNA) have tumor suppressive and oncogenic potential in human cancer, but whether and how miRNAs control cell cycle progression is not understood. To address this question, we carried out a comprehensive analysis of miRNA expression during serum stimulation of quiescent human cells. Time course analyses revealed that four miRNAs are up-regulated and >100 miRNAs are down regulated, as cells progress beyond the G(1)-S phase transition. We analyzed the function of two up-regulated miRNAs (miR-221 and miR-222) that are both predicted to target the cell growth suppressive cyclin-dependent kinase inhibitors p27 and p57. Our results show that miR-221 and miR-222 both directly target the 3' untranslated regions of p27 and p57 mRNAs to reduce reporter gene expression, as well as diminish p27 and p57 protein levels. Functional studies show that miR-221 and miR-222 prevent quiescence when elevated during growth factor deprivation and induce precocious S-phase entry, thereby triggering cell death. Thus, the physiologic up-regulation of miR-221 and miR-222 is tightly linked to a cell cycle checkpoint that ensures cell survival by coordinating competency for initiation of S phase with growth factor signaling pathways that stimulate cell proliferation. PMID- 18413745 TI - Manganese superoxide dismutase modulates hypoxia-inducible factor-1 alpha induction via superoxide. AB - Hypoxia-inducible factor 1 (HIF-1) is a transcription factor that plays an important role in O(2) homeostasis. Numerous observations suggest that changes in reactive oxygen species affect HIF-1 alpha stabilization and HIF-1 alpha transcriptional activation in many cell types. The antioxidant enzyme manganese superoxide dismutase (MnSOD) modulates the cellular redox environment by converting superoxide (O(2)(*-)) to hydrogen peroxide and dioxygen. Previous results from our group have shown that overexpression of MnSOD in MCF-7 cells alters stabilization of HIF-1 alpha under hypoxic conditions; however, the underlying mechanism(s) is not known. Here, we tested the hypothesis that MnSOD regulates the expression of HIF-1 alpha by modulating the steady-state level of O(2)(*-). We found that decreasing MnSOD with small interfering RNA in MCF-7 cells resulted in (a) an associated increase in the hypoxic accumulation of HIF-1 alpha immunoreactive protein, (b) a significant increase in the levels of O(2)(* ) (P < 0.01), but (c) no significant change in the steady-state level of H(2)O(2). Removal of O(2)(*-) using spin traps (alpha-4-pyridyl-1-oxide-N-tert butylnitrone and 5,5-dimethyl-1-pyrroline N-oxide) or the O(2)(*-) scavenger Tempol or an SOD mimic (AEOL10113) resulted in a decrease in HIF-1 alpha protein, consistent with the hypothesis that O(2)(*-) is an important molecular effector responsible for hypoxic stabilization of HIF-1 alpha. The evidence from both genetic and pharmaceutical manipulation is consistent with our hypothesis that O(2)(*-) can contribute to the stabilization of HIF-1 alpha. PMID- 18413746 TI - Negative regulation of YAP by LATS1 underscores evolutionary conservation of the Drosophila Hippo pathway. AB - The Hippo pathway defines a novel signaling cascade regulating cell proliferation and survival in Drosophila, which involves the negative regulation of the transcriptional coactivator Yorkie by the kinases Hippo and Warts. We have recently shown that the human ortholog of Yorkie, YAP, maps to a minimal amplification locus in mouse and human cancers, and that it mediates dramatic transforming activity in MCF10A primary mammary epithelial cells. Here, we show that LATS proteins (mammalian orthologs of Warts) interact directly with YAP in mammalian cells and that ectopic expression of LATS1, but not LATS2, effectively suppresses the YAP phenotypes. Furthermore, shRNA-mediated knockdown of LATS1 phenocopies YAP overexpression. Because this effect can be suppressed by simultaneous YAP knockdown, it suggests that YAP is the primary target of LATS1 in mammalian cells. Expression profiling of genes induced by ectopic expression of YAP or by knockdown of LATS1 reveals a subset of potential Hippo pathway targets implicated in epithelial-to-mesenchymal transition, suggesting that this is a key feature of YAP signaling in mammalian cells. PMID- 18413747 TI - Down-regulation of caspase-2 by rottlerin via protein kinase C-delta-independent pathway. AB - Protein kinase C-delta (PKC delta) plays an important role in DNA damage-induced apoptosis. We have previously shown that the PKC delta inhibitor rottlerin protects against cisplatin-induced apoptosis acting upstream of caspase-9. In the present study, we have investigated if rottlerin regulates caspase-2 activation. Knockdown of caspase-2 by siRNA inhibited processing of apical caspase-9 and caspase-8, whereas depletion of caspase-9 had little effect on caspase-2 processing. Rottlerin inhibited activation and processing of caspase-9 and caspase-8 and cleavage of poly(ADP)ribose polymerase. We made a novel observation that rottlerin induced down-regulation of caspase-2 but not of caspase-3, caspase 7, caspase-8, or caspase-9. Pharmacologic inhibitors of PKC, such as Go 6983 and bisindolylmaleimide, or depletion of PKC delta by siRNA had no effect on the down regulation of caspase-2 by rottlerin. The proteasome inhibitor MG132 reversed caspase-2 down-regulation by rottlerin, whereas calpain inhibitor had no effect. These results suggest that rottlerin induces down-regulation of caspase-2 via PKC delta-independent but ubiquitin proteasome-mediated pathway. Furthermore, down regulation of caspase-2 by rottlerin can explain its antiapoptotic function during DNA damage-induced apoptosis. PMID- 18413748 TI - Targeting CD24 for treatment of colorectal and pancreatic cancer by monoclonal antibodies or small interfering RNA. AB - CD24 is a potential oncogene reported to be overexpressed in a large variety of human malignancies. We have shown that CD24 is overexpressed in 90% of colorectal tumors at a fairly early stage in the multistep process of carcinogenesis. Anti CD24 monoclonal antibodies (mAb) induce a significant growth inhibition in colorectal and pancreatic cancer cell lines that express the protein. This study is designed to investigate further the effects of CD24 down-regulation using mAb or small interfering RNA in vitro and in vivo. Western blot analysis showed that anti-CD24 mAb induced CD24 protein down-regulation through lysosomal degradation. mAb augmented growth inhibition in combination with five classic chemotherapies. Xenograft models in vivo showed that tumor growth was significantly reduced in mAb-treated mice. Similarly, stable growth inhibition of cancer cell lines was achieved by down-regulation of CD24 expression using short hairpin RNA (shRNA). The produced clones proliferated more slowly, reached lower saturation densities, and showed impaired motility. Most importantly, down-regulation of CD24 retarded tumorigenicity of human cancer cell lines in nude mice. Microarray analysis revealed a similar pattern of gene expression alterations when cells were subjected to anti-CD24 mAb or shRNA. Genes in the Ras pathway, mitogen-activated protein kinase, or BCL-2 family and others of oncogenic association were frequently down-regulated. As a putative new oncogene that is overexpressed in gastrointestinal malignancies early in the carcinogenesis process, CD24 is a potential target for early intervention in the prevention and treatment of cancer. PMID- 18413749 TI - The mitochondrial uncoupling protein-2 promotes chemoresistance in cancer cells. AB - Cancer cells acquire drug resistance as a result of selection pressure dictated by unfavorable microenvironments. This survival process is facilitated through efficient control of oxidative stress originating from mitochondria that typically initiates programmed cell death. We show this critical adaptive response in cancer cells to be linked to uncoupling protein-2 (UCP2), a mitochondrial suppressor of reactive oxygen species (ROS). UCP2 is present in drug-resistant lines of various cancer cells and in human colon cancer. Overexpression of UCP2 in HCT116 human colon cancer cells inhibits ROS accumulation and apoptosis after exposure to chemotherapeutic agents. Tumor xenografts of UCP2-overexpressing HCT116 cells retain growth in nude mice receiving chemotherapy. Augmented cancer cell survival is accompanied by altered NH(2)-terminal phosphorylation of the pivotal tumor suppressor p53 and induction of the glycolytic phenotype (Warburg effect). These findings link UCP2 with molecular mechanisms of chemoresistance. Targeting UCP2 may be considered a novel treatment strategy for cancer. PMID- 18413750 TI - Targeting endoplasmic reticulum stress and Akt with OSU-03012 and gefitinib or erlotinib to overcome resistance to epidermal growth factor receptor inhibitors. AB - Preexisting and acquired resistance to epidermal growth factor receptor (EGFR) inhibitors limits their clinical usefulness in patients with advanced non-small cell lung cancer (NSCLC). This study characterizes the efficacy and mechanisms of the combination of gefitinib or erlotinib with OSU-03012, a celecoxib-derived antitumor agent, to overcome EGFR inhibitor resistance in three NSCLC cell lines, H1155, H23, and A549. The OSU-03012/EGFR inhibitor combination induced pronounced apoptosis in H1155 and H23 cells, but not in A549 cells, suggesting a correlation between drug sensitivity and basal phospho-Akt levels independently of EGFR expression status. Evidence indicates that this combination facilitates apoptosis through both Akt signaling inhibition and up-regulation of endoplasmic reticulum (ER) stress-induced, GADD153-mediated pathways. For example, ectopic expression of constitutively active Akt significantly attenuated the inhibitory effect on cell survival, and small interfering RNA-mediated knockdown of GADD153 protected cells from undergoing apoptosis in response to drug cotreatments. Furthermore, the OSU-03012/EGFR inhibitor combination induced GADD153-mediated up-regulation of death receptor 5 expression and subsequent activation of the extrinsic apoptosis pathway. It is noteworthy that the ER stress response induced by this combination was atypical in that the cytoprotective pathway was not engaged. In addition, in vivo suppression of tumor growth and modulation of intratumoral biomarkers were observed in a H1155 tumor xenograft model in nude mice. These data suggest that the concomitant modulation of Akt and ER stress pathways with the OSU-03012/EGFR inhibitor combination represents a unique approach to overcoming EGFR inhibitor resistance in NSCLC and perhaps other types of cancer with elevated basal Akt activities. PMID- 18413751 TI - Synthetic lethality by lentiviral short hairpin RNA silencing of thymidylate kinase and doxorubicin in colon cancer cells regardless of the p53 status. AB - Intracellular supply of dTTP is a highly regulated process and has been a key target for chemotherapeutic drug development. Thymidylate kinase (TMPK) is the key enzyme for dTTP formation in both de novo and salvage pathways. In this study, we used lentiviral-based small hairpin RNA to silence TMPK expression in p53(+/+) and p53(-/-) HCT-116 colon cancer cells. This approach was sufficient to decrease the dTTP pool gradually without affecting p53 expression and generating cytotoxicity. TMPK knockdown significantly increased doxorubicin sensitivity dramatically in p53-proficient, p53-null HCT-116, and LoVo colon cancer cells. The decrease in the dTTP pool using this approach augmented the DNA damage response and enhanced apoptotic induction after exposure to low-dose doxorubicin, leading to cell death. In contrast, silencing of thymidylate synthase which blocks the de novo pathway was incapable of sensitizing p53-null HCT-116 cells to doxorubicin-induced apoptosis because of the compensation by the salvage pathway. Our results suggest the lentiviral delivery of small hairpin RNA targeting TMPK in combination with a low dose of doxorubicin as a new approach to kill colon cancer cells regardless of p53 status. PMID- 18413752 TI - Coordinated epidermal growth factor receptor pathway gene overexpression predicts epidermal growth factor receptor inhibitor sensitivity in pancreatic cancer. AB - The epidermal growth factor receptor (EGFR) inhibitor erlotinib is approved for treatment of pancreatic cancer but the overall activity is minimal, and known predictive factors for EGFR inhibitor efficacy are infrequent in this disease. We tested the hypothesis that global activation of the EGFR pathway is predictive of EGFR inhibitor efficacy. Pancreatic cancer tumors directly xenografted at surgery were treated with the EGFR inhibitors erlotinib and cetuximab and analyzed for biological features. Two of 10 tumors were sensitive, and by global gene expression profiling with gene set enrichment analysis, the EGFR pathway was highly expressed in sensitive compared with resistant tumors. The core gene components driving EGFR pathway overexpression were pathway ligands and positive effectors. In a prospective validation, the EGFR pathway-based signature correctly predicted anti-EGFR treatment response in eight additional tumors and was not predictive of response to gemcitabine and CI1040 (a MEK inhibitor). Analysis of EGFR, KRAS, and PIK3CA mutations and gene amplification by fluorescence in situ hybridization and multiplex ligation-dependent probe amplification showed that none of these genetic abnormalities were neither predictive nor responsible for the EGFR pathway activation. Coordinated overexpression of the EGFR pathway predicts susceptibility to EGFR inhibitors in pancreatic cancer. These results suggest a phenomenon of pathway addiction and support the value of unbiased system biology approaches in drug development. PMID- 18413753 TI - NVP-AUY922: a novel heat shock protein 90 inhibitor active against xenograft tumor growth, angiogenesis, and metastasis. AB - We describe the biological properties of NVP-AUY922, a novel resorcinylic isoxazole amide heat shock protein 90 (HSP90) inhibitor. NVP-AUY922 potently inhibits HSP90 (K(d) = 1.7 nmol/L) and proliferation of human tumor cells with GI(50) values of approximately 2 to 40 nmol/L, inducing G(1)-G(2) arrest and apoptosis. Activity is independent of NQO1/DT-diaphorase, maintained in drug resistant cells and under hypoxic conditions. The molecular signature of HSP90 inhibition, comprising induced HSP72 and depleted client proteins, was readily demonstrable. NVP-AUY922 was glucuronidated less than previously described isoxazoles, yielding higher drug levels in human cancer cells and xenografts. Daily dosing of NVP-AUY922 (50 mg/kg i.p. or i.v.) to athymic mice generated peak tumor levels at least 100-fold above cellular GI(50). This produced statistically significant growth inhibition and/or regressions in human tumor xenografts with diverse oncogenic profiles: BT474 breast tumor treated/control, 21%; A2780 ovarian, 11%; U87MG glioblastoma, 7%; PC3 prostate, 37%; and WM266.4 melanoma, 31%. Therapeutic effects were concordant with changes in pharmacodynamic markers, including induction of HSP72 and depletion of ERBB2, CRAF, cyclin-dependent kinase 4, phospho-AKT/total AKT, and hypoxia-inducible factor-1alpha, determined by Western blot, electrochemiluminescent immunoassay, or immunohistochemistry. NVP-AUY922 also significantly inhibited tumor cell chemotaxis/invasion in vitro, WM266.4 melanoma lung metastases, and lymphatic metastases from orthotopically implanted PC3LN3 prostate carcinoma. NVP-AUY922 inhibited proliferation, chemomigration, and tubular differentiation of human endothelial cells and antiangiogenic activity was reflected in reduced microvessel density in tumor xenografts. Collectively, the data show that NVP-AUY922 is a potent, novel inhibitor of HSP90, acting via several processes (cytostasis, apoptosis, invasion, and angiogenesis) to inhibit tumor growth and metastasis. NVP-AUY922 has entered phase I clinical trials. PMID- 18413754 TI - Bone marrow X kinase-mediated signal transduction in irradiated vascular endothelium. AB - Radiation-induced activation of the phosphatidyl inositol-3 kinase/Akt signal transduction pathway requires Akt binding to phosphatidyl-inositol phosphates (PIP) on the cell membrane. The tyrosine kinase bone marrow X kinase (Bmx) binds to membrane-associated PIPs in a manner similar to Akt. Because Bmx is involved in cell growth and survival pathways, it could contribute to the radiation response within the vascular endothelium. We therefore studied Bmx signaling within the vascular endothelium. Bmx was activated rapidly in response to clinically relevant doses of ionizing radiation. Bmx inhibition enhanced the efficacy of radiotherapy in endothelial cells as well as tumor vascular endothelium in lung cancer tumors in mice. Retroviral shRNA knockdown of Bmx protein enhanced human umbilical vascular endothelial cell (HUVEC) radiosensitization. Furthermore, pretreatment of HUVEC with a pharmacologic inhibitor of Bmx, LFM-A13, produced significant radiosensitization of endothelial cells as measured by clonogenic survival analysis and apoptosis as well as functional assays including cell migration and tubule formation. In vivo, LFM A13, when combined with radiation, resulted in significant tumor microvascular destruction as well as enhanced tumor growth delay. Bmx therefore represents a molecular target for the development of novel radiosensitizing agents. PMID- 18413755 TI - NOV-002, a glutathione disulfide mimetic, as a modulator of cellular redox balance. AB - NOV-002 is a novel glutathione disulfide mimetic that when administered in combination with standard chemotherapeutic regimens has resulted in increased efficacy (survival, tumor response) and improved tolerance to chemotherapy (e.g., hematologic recovery) in advanced non-small cell lung cancer patients. We show that NOV-002, which is not cytotoxic as a single agent, generated time- and concentration-dependent oxidative signals at the cell surface (reduction in protein thiols) and intracellularly [altered oxidized glutathione (GSSG) and reduced glutathione levels and ratio; increased reactive oxygen species] in the premyeloid HL-60 cell line and that this was associated with an increase in S glutathionylation of cell proteins, particularly actin. Commensurate with these effects, NOV-002 activated p38, c-Jun-NH(2)-kinase, and extracellular signal regulated kinase and caused a dose-dependent increase in phosphorylation of three proteins that have previously been linked with hematopoiesis, AKT, JAK2, and STAT5. The effect of NOV-002 on enzymes involved in glutathione metabolism was evaluated. Relative to oxidized glutathione, NOV-002 was an equivalent substrate for glutathione reductase and was an inhibitor of protein disulfide isomerase, one of the components of the redox-sensitive unfolded protein response pathway. These redox-stimulated cell signaling actions occurred in the context of increased HL-60 cell proliferation after treatment with NOV-002. Overall, the pleiotropic pharmacologic effects of NOV-002 can be attributed to the GSSG component of the drug, and modulation of cellular redox balance is a feature central to the mechanism of action of NOV-002. Such modulation may underlie its clinical actions, including hematologic recovery and immunostimulation in the face of chemosuppression. PMID- 18413756 TI - Imaging transgene activity in vivo. AB - The successful translation of gene therapy for clinical application will require the assessment of transgene activity as a measure of the biological function of a therapeutic transgene. Although current imaging permits the noninvasive detection of transgene expression, the critical need for quantitative imaging of the action of the expressed transgene has not been met. In vivo magnetic resonance spectroscopic imaging (MRSI) was applied to quantitatively delineate both the concentration and activity of a cytosine deaminase-uracil phosphoribosyltransferase (CD-UPRT) fusion enzyme expressed from a transgene. MRSI enabled the generation of anatomically accurate maps of the intratumoral heterogeneity in fusion enzyme activity. We observed an excellent association between the CD-UPRT concentration and activity and the percentage of CD-UPRT(+) cells. Moreover, the regional levels of UPRT activity, as measured by imaging, correlated well with the biological affect of the enzyme. This study presents a translational imaging paradigm for precise, in vivo measurements of transgene activity with potential applications in both preclinical and clinical settings. PMID- 18413757 TI - Targeting a tumor-specific laminin domain critical for human carcinogenesis. AB - Laminin-332 is critical for squamous cell carcinoma (SCC) tumorigenesis, but targeting it for cancer therapy has been unachievable due to key role of laminin 332 in promoting tissue integrity. Here, we show that a portion of laminin-332, termed G45, which is proteolytically removed and absent in normal tissues, is prominently expressed in most human SCC tumors and plays an important role in human SCC tumorigenesis. Primary human keratinocytes lacking G45 (DeltaG45) showed alterations of basal receptor organization, impaired matrix deposition, and increased migration. After SCC transformation, the absence of G45 domain in DeltaG45 cells was associated with deficient extracellular signal-regulated kinase and phosphotidylinositol 3-kinase (PI3K) pathway activation, impaired invasion, deficient metalloproteinase activity, and absent tumorgenicity in vivo. Expression of G45 or activated PI3K subunit in DeltaG45 cells reversed these abnormalities. G45 antibody treatment induced SCC tumor apoptosis, decreased SCC tumor proliferation, and markedly impaired human SCC tumorigenesis in vivo without affecting normal tissue adhesion. These results show a remarkable selectivity of expression and function for laminin-332 G45 in human SCC tumorigenesis and implicate it as a specific target for anticancer therapy. PMID- 18413758 TI - Soft tissue sarcoma cells are highly sensitive to AKT blockade: a role for p53 independent up-regulation of GADD45 alpha. AB - The AKT signaling pathway is activated in soft tissue sarcoma (STS). However, AKT blockade has not yet been studied as a potential targeted therapeutic approach. Here, we examined the in vitro and in vivo effects of AKT inhibition in STS cells. Western blot analysis was used to evaluate the expression of AKT pathway components and the effect of AKT stimulation and inhibition on their phosphorylation. Cell culture assays were used to assess the effect of AKT blockade (using a phosphatidylinositol 3-kinase inhibitor and a specific AKT inhibitor) on STS cell growth, cell cycle, and apoptosis. Oligoarrays were used to determine gene expression changes in response to AKT inhibition. Reverse transcription-PCR was used for array validation. Specific small inhibitory RNA was used to knockdown GADD45 alpha. Human STS xenografts in nude mice were used for in vivo studies, and immunohistochemistry was used to assess the effect of treatment on GADD45 alpha expression, proliferation, and apoptosis. Multiple STS cell lines expressed activated AKT. AKT inhibition decreased STS downstream target phosphorylation and growth in vitro; G(2) cell cycle arrest and apoptosis were also observed. AKT inhibition induced GADD45 alpha mRNA and protein expression in all STS cells treated independent of p53 mutational status. GADD45 alpha knockdown attenuated the G(2) arrest induced by AKT inhibition. In vivo, AKT inhibition led to decreased STS xenograft growth. AKT plays a critical role in survival and proliferation of STS cells. Modulation of AKT kinase activity may provide a novel molecularly based strategy for STS-targeted therapies. PMID- 18413759 TI - New derivatives of vitamin B12 show preferential targeting of tumors. AB - Rapidly growing cells show an increased demand for nutrients and vitamins. The objective of our work is to exploit the supply route of vitamin B12 to deliver new derivatives of this vital vitamin to hyperproliferative cells. To date, radiolabeled ((57)Co and (111)In) vitamin B12 derivatives showed labeling of tumor tissue but also undesired high accumulation of radioactivity in normal tissue. By abolishing the interaction of a tailored vitamin B12 derivative to its transport protein transcobalamin II and therefore interrupting transcobalamin II receptor and megalin mediated uptake in normal tissue, preferential accumulation of a radiolabeled vitamin in cancer tissue could be accomplished. We identified transcobalamin I on tumors as a possible new receptor for this preferential accumulation of vitamin-mediated targeting. The low systemic distribution of radioactivity and the high tumor to blood ratio opens the possibility of a more successful clinical application of vitamin B12 for imaging or therapy. PMID- 18413760 TI - Peroxisome proliferator-activated receptor gamma-mediated up-regulation of syndecan-1 by n-3 fatty acids promotes apoptosis of human breast cancer cells. AB - Diets enriched in n-3 polyunsaturated fatty acids (n-3 PUFA) may protect against breast cancer but biochemical mechanisms are unclear. Our studies showed that the n-3 fatty acid docosahexaenoic acid (DHA) up-regulated syndecan-1 (SDC-1) in human breast cancer cells, and we tested the hypothesis that DHA-mediated up regulation of SDC-1 induces apoptosis. DHA was delivered to MCF-7 cells by n-3 PUFA-enriched low-density lipoproteins (LDL) or by albumin in the presence or absence of SDC-1 small interfering RNA. The n-3 PUFA induced apoptosis, which was blocked by SDC-1 silencing. We also confirmed that SDC-1 up-regulation and apoptosis promotion by n-3 PUFA was mediated by peroxisome proliferator-activated receptor gamma (PPAR gamma). Using a luciferase gene driven by either a PPAR response element or a DR-1 site present in the SDC-1 promoter, reporter activities were enhanced by n-3 LDL, DHA, and PPAR gamma agonist, whereas activity of a luciferase gene placed downstream of a mutant DR-1 site was unresponsive. Cotransfection with dominant-negative PPAR gamma DNA eliminated the increase in luciferase activity. These data provide strong evidence that SDC-1 is a molecular target of n-3 PUFA in human breast cancer cells through activation of PPAR gamma and that n-3 PUFA-induced apoptosis is mediated by SDC-1. This provides a novel mechanism for the chemopreventive effects of n-3 PUFA in breast cancer. PMID- 18413761 TI - Triterpenoid CDDO-methyl ester inhibits the Janus-activated kinase-1 (JAK1)- >signal transducer and activator of transcription-3 (STAT3) pathway by direct inhibition of JAK1 and STAT3. AB - The C-28 methyl ester of the oleane triterpenoid 2-cyano-3,12-dioxooleana-1,9 dien-28-oic acid (CDDO-Me) induces apoptosis of human cancer cells by disrupting redox balance and is in clinical trials. CDDO-Me contains alpha,beta-unsaturated carbonyl groups that form reversible adducts with thiol nucleophiles. The present studies show that CDDO-Me blocks interleukin-6 (IL-6)-induced and constitutive activation of the Janus-activated kinase 1 (JAK1) in cells. In support of a direct mechanism, CDDO-Me forms adducts with JAK1 at Cys(1077) in the kinase domain and inhibits JAK1 activity. In concert with these results, CDDO-Me blocked IL-6-induced and constitutive activation of signal transducer and activator of transcription 3 (STAT3). Moreover, we show that CDDO-Me (a) binds directly to STAT3 by a mechanism dependent on the alkylation of Cys(259) and (b) inhibits the formation of STAT3 dimers. These findings indicate that CDDO-Me inhibits activation of the JAK1-->STAT3 pathway by forming adducts with both JAK1 and STAT3. PMID- 18413762 TI - Apoptotic activity and mechanism of 2-cyano-3,12-dioxoolean-1,9-dien-28-oic-acid and related synthetic triterpenoids in prostate cancer. AB - Synthetic triterpenoids 2-cyano-3, 12-dioxooleana-1, 9-(11)-dien-28-oic acid (CDDO) and CDDO-Me (CDDO-methyl ester) have entered clinical trials for cancer. We determined that CDDO analogues at submicromolar concentrations induce apoptosis of cultured prostate cancer cell lines, LNCaP, ALVA31, Du145, PC3, and PPC1, with lethal dose 50% approximately 1 micromol/L for CDDO-Me and an imidazole analogue (CDDO-Im). These compounds induced apoptosis of prostate cancer cells as characterized by cleavage of caspase-3, caspase-7, caspase-8, caspase-9, caspase-10, BID, and poly(ADP)ribose polymerase and by dependence on caspase activity. Moreover, triterpenoid-induced cell death was abolished by caspase-8-targeting small interfering (si) RNA. To explore the mechanism(s) involved in caspase-8 activation, we examined cell surface expression of death receptor (DR)4 and DR5 after triterpenoid treatment. Cell surface DR4 and DR5 expression was significantly up-regulated by CDDO or CDDO-Im but not by CDDO-Me. DR4 and DR5 knockdown with siRNA significantly inhibited apoptosis induced by CDDO and CDDO-Im but had no effect on CDDO-Me-induced killing, suggesting that CDDO and CDDO-Im induce apoptosis by a different mechanism than CDDO-Me. In addition to activating the caspase-8-dependent extrinsic apoptosis pathway, we observed that Bcl-X(L) overexpression inhibited triterpenoid-mediated killing of prostate cancer cell line Du145, suggesting that the intrinsic pathway (via mitochondria) also participates in triterpenoid-mediated killing. In vivo antitumor activity of CDDO-Me was shown using a Du145 tumor xenograft model in nude rats. Altogether, these findings suggest CDDO and related synthetic triterpenoids should be further evaluated as potential novel therapeutics for hormone refractory prostate cancers. PMID- 18413763 TI - A new pharmacologic action of CCI-779 involves FKBP12-independent inhibition of mTOR kinase activity and profound repression of global protein synthesis. AB - The mammalian target of rapamycin (mTOR) inhibitor CCI-779 (temsirolimus) is a recently Food and Drug Administration-approved anticancer drug with efficacy in certain solid tumors and hematologic malignancies. In cell culture studies, CCI 779 at the commonly used nanomolar concentrations generally confers a modest and selective antiproliferative activity. Here, we report that, at clinically relevant low micromolar concentrations, CCI-779 completely suppressed proliferation of a broad panel of tumor cells. This "high-dose" drug effect did not require FKBP12 and correlated with an FKBP12-independent suppression of mTOR signaling. An FKBP12-rapamycin binding domain (FRB) binding-deficient rapamycin analogue failed to elicit both the nanomolar and micromolar inhibitions of growth and mTOR signaling, implicating FRB binding in both actions. Biochemical assays indicated that CCI-779 and rapamycin directly inhibited mTOR kinase activity with IC(50) values of 1.76 +/- 0.15 and 1.74 +/- 0.34 micromol/L, respectively. Interestingly, a CCI-779-resistant mTOR mutant (mTOR-SI) displayed an 11-fold resistance to the micromolar CCI-779 in vitro (IC(50), 20 +/- 3.4 micromol/L) and conferred a partial protection in cells exposed to micromolar CCI-779. Treatment of cancer cells with micromolar but not nanomolar concentrations of CCI-779 caused a marked decline in global protein synthesis and disassembly of polyribosomes. The profound inhibition of protein synthesis was accompanied by rapid increase in the phosphorylation of translation elongation factor eEF2 and the translation initiation factor eIF2 alpha. These findings suggest that high dose CCI-779 inhibits mTOR signaling through an FKBP12-independent mechanism that leads to profound translational repression. This distinctive high-dose drug effect could be directly related to the antitumor activities of CCI-779 and other rapalogues in human cancer patients. PMID- 18413764 TI - BH3 mimetic ABT-737 potentiates TRAIL-mediated apoptotic signaling by unsequestering Bim and Bak in human pancreatic cancer cells. AB - Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) has been shown to induce mitochondrial apoptotic signaling that can be negatively regulated by prosurvival Bcl-2 proteins. ABT-737 is a small-molecule BH3 mimetic that binds to and antagonizes Bcl-2/Bcl-x(L) but not Mcl-1. We show that ABT-737 can synergistically enhance TRAIL-mediated cytotoxicity in human pancreatic cancer cell lines. ABT-737 was shown to enhance TRAIL-induced apoptosis as shown by DNA fragmentation, activation of caspase-8 and Bid, and cleavage of caspase-3 and poly(ADP-ribose) polymerase. A Bax conformational change induced by TRAIL was enhanced by ABT-737. ABT-737 disrupted the interaction of Bak with Bcl-x(L) in both cell lines. Furthermore, ABT-737 untethered the proapoptotic BH3-only protein Bim from its sequestration by Bcl-x(L) or Bcl-2. Bim small hairpin RNA (shRNA) was shown to attenuate caspase-3 cleavage and to reduce the cytotoxic effects of TRAIL plus ABT-737 compared with shRNA control cells. Finally, Mcl-1 shRNA potentiated caspase-3 cleavage by ABT-737 and enhanced its cytotoxic effects. Taken together, ABT-737 augments TRAIL-induced cell killing by unsequestering Bim and Bak and enhancing a Bax conformational change induced by TRAIL. These findings suggest a novel strategy to enhance cross-talk between the extrinsic and intrinsic apoptotic pathways to improve therapeutic efficacy against pancreatic cancer. PMID- 18413765 TI - Containment of tumor-colonizing bacteria by host neutrophils. AB - Administration of facultative anaerobic bacteria like Salmonella typhimurium, Shigella flexneri, and Escherichia coli to tumor-bearing mice leads to a preferential accumulation and proliferation of the microorganisms within the solid tumor. Until now, all known tumor-targeting bacteria have shown poor dissemination inside the tumors. They accumulate almost exclusively in large necrotic areas and spare a rim of viable tumor cells. Interestingly, the bacteria containing necrotic region is separated from viable tumor cells by a barrier of host neutrophils that have immigrated into the tumor. We here report that depletion of host neutrophils results in a noticeably higher total number of bacteria in the tumor and that bacteria were now also able to migrate into vital tumor tissue. Most remarkably, an increase in the size of the necrosis was observed, and complete eradication of established tumors could be observed under these conditions. Thus, bacteria-mediated tumor therapy can be amplified by depletion of host neutrophils. PMID- 18413766 TI - Redirecting specificity of T-cell populations for CD19 using the Sleeping Beauty system. AB - Genetic modification of clinical-grade T cells is undertaken to augment function, including redirecting specificity for desired antigen. We and others have introduced a chimeric antigen receptor (CAR) to enable T cells to recognize lineage-specific tumor antigen, such as CD19, and early-phase human trials are currently assessing safety and feasibility. However, a significant barrier to next-generation clinical studies is developing a suitable CAR expression vector capable of genetically modifying a broad population of T cells. Transduction of T cells is relatively efficient but it requires specialized manufacture of expensive clinical grade recombinant virus. Electrotransfer of naked DNA plasmid offers a cost-effective alternative approach, but the inefficiency of transgene integration mandates ex vivo selection under cytocidal concentrations of drug to enforce expression of selection genes to achieve clinically meaningful numbers of CAR(+) T cells. We report a new approach to efficiently generating T cells with redirected specificity, introducing DNA plasmids from the Sleeping Beauty transposon/transposase system to directly express a CD19-specific CAR in memory and effector T cells without drug selection. When coupled with numerical expansion on CD19(+) artificial antigen-presenting cells, this gene transfer method results in rapid outgrowth of CD4(+) and CD8(+) T cells expressing CAR to redirect specificity for CD19(+) tumor cells. PMID- 18413767 TI - Interleukin-15/interleukin-15R alpha complexes promote destruction of established tumors by reviving tumor-resident CD8+ T cells. AB - Tumors often escape immune-mediated destruction by suppressing lymphocyte infiltration or effector function. New approaches are needed that overcome this suppression and thereby augment the tumoricidal capacity of tumor-reactive lymphocytes. The cytokine interleukin-15 (IL-15) promotes proliferation and effector capacity of CD8(+) T cells, natural killer (NK) cells, and NKT cells; however, it has a short half-life and high doses are needed to achieve functional responses in vivo. The biological activity of IL-15 can be dramatically increased by complexing this cytokine to its soluble receptor, IL-15R alpha. Here, we report that in vivo delivery of IL-15/IL-15R alpha complexes triggers rapid and significant regression of established solid tumors in two murine models. Despite a marked expansion of IL-2/IL-15R beta(+) cells in lymphoid organs and peripheral blood following treatment with IL-15/IL-15R alpha complexes, the destruction of solid tumors was orchestrated by tumor-resident rather than newly infiltrating CD8(+) T cells. Our data provide novel insights into the use of IL-15/IL-15R alpha complexes to relieve tumor-resident T cells from functional suppression by the tumor microenvironment and have significant implications for cancer immunotherapy and treatment of chronic infections. PMID- 18413768 TI - Modeling the CD8+ T effector to memory transition in adoptive T-cell antitumor immunotherapy. AB - Adoptive T-cell therapy with CD8(+) CTLs is often characterized by poor persistence of the transferred T cells and limited effector responses. Improved persistence and therapeutic efficacy have been noted when antigen-activated CD8(+) T cells express properties of memory cells. The current study was undertaken to more precisely characterize the development of memory-like CD8(+) T cells from short-term CTLs in vitro and upon transfer in vivo, including their antitumor activity. Ovalbumin (OVA)-specific OT-I CTLs acquired phenotypic and functional properties of memory cells 2 to 3 days later either by lowering the concentration of antigen to a level that does not support primary responses and providing a survival signal through transgenic Bcl-2 in vitro or simply by transferring early day 3 CTLs to antigen-free lymphoid-replete mice. In lymphoid replete mice, established OVA-expressing E.G7 tumor was rejected by short-term CTLs that simultaneously acquired memory-like properties in secondary lymphoid tissues, where tumor antigen level remained low. Collectively, these data indicate that CTLs readily converted to memory-like cells upon lowering antigen to a concentration that selectively supports memory responses and suggest that such conversion predicts successful adoptive immunotherapy. PMID- 18413769 TI - Loss of CTL function among high-avidity tumor-specific CD8+ T cells following tumor infiltration. AB - A major problem in generating effective antitumor CTL responses is that most tumors express self-antigens to which the immune system is rendered unresponsive due to mechanisms of self-tolerance induction. CTL precursors (CTLp) expressing high-affinity T-cell receptors (TCR) are often functionally deleted from the repertoire, leaving a residual repertoire of CTLp having only low-affinity TCR. Furthermore, even when unique antigens are expressed, their presentation by dendritic cells (DC) may predispose to peripheral tolerance induction rather than the establishment of CTL responses that kill tumor cells. In this study, we examined both high-avidity (CL4) and low-avidity (CL1) CD8(+) T-cell responses to a murine renal carcinoma expressing, as a neoantigen, high and low levels of the hemagglutinin (HA) protein from influenza virus A/PR/8 H1N1 (PR8; RencaHA(high) and RencaHA(low)). Our data show that, following encounter with K(d)HA epitopes cross-presented by bone marrow-derived DC, low-avidity CL1 cells become tolerized within tumor-draining lymph nodes (TDLN), and in mice bearing either RencaHA(high) or RencaHA(low) tumors, very few form tumor-infiltrating lymphocytes (TIL). In marked contrast, high-avidity CL4 cells differentiate into effector CTL within the TDLN of mice bearing either RencaHA(high) or RencaHA(low) tumors, and although they form TIL in both tumors, they lose CTL effector function. Critically, these results show that anticancer therapies involving either adoptive transfer of high-avidity tumor-specific CTL populations or targeting of preexisting tumor antigen-specific memory CD8(+) T cells could fail due to the fact that CTL effector function is lost following tumor infiltration. PMID- 18413770 TI - The regulatory T cell-associated transcription factor FoxP3 is expressed by tumor cells. AB - FoxP3 is a member of the forkhead family of transcription factors critically involved in the development and function of CD25(+) regulatory T cells (Treg). Until recently, FoxP3 expression was thought to be restricted to the T-cell lineage. However, using immunohistochemistry and flow cytometric analysis of human melanoma tissue, we detected FoxP3 expression not only in the tumor infiltrating Treg but also in the melanoma cells themselves. FoxP3 is also widely expressed by established human melanoma cell lines (as determined by flow cytometry, PCR, and Western blot), as well as cell lines derived from other solid tumors. Normal B cells do not express FoxP3; however, expression could be induced after transformation with EBV in vitro and in vivo, suggesting that malignant transformation of healthy cells can induce FoxP3. In addition, a FOXP3 mRNA variant lacking exons 3 and 4 was identified in tumor cell lines but was absent from Treg. Interestingly, this alternative splicing event introduces a translation frame-shift that is predicted to encode a novel protein. Together, our results show that FoxP3, a key regulator of immune suppression, is not only expressed by Treg but also by melanoma cells, EBV-transformed B cells, and a wide variety of tumor cell lines. PMID- 18413771 TI - Synthetic CD4+ T cell-targeted antigen-presenting cells elicit protective antitumor responses. AB - CD4(+) helper T cells are critical for protective immune responses and yet suboptimally primed in response to tumors. Cell-based vaccination strategies are under evaluation in clinical trials but limited by the need to derive antigen presenting cells (APC) from patients or compatible healthy donors. To overcome these limitations, we developed CD4(+) T cell-targeted synthetic microbead-based artificial APC (aAPC) and used them to activate CD4(+) T lymphocytes specific for a tumor-associated model antigen (Ag) directly from the naive repertoire. In vitro, aAPC specifically primed Ag-specific CD4(+) T cells that were activated to express high levels of CD44, produced mainly interleukin 2, and could differentiate into Th1-like or Th2-like cells in combination with polarizing cytokines. I.v. administration of aAPC led to Ag-specific CD4(+) T-cell activation and proliferation in secondary lymphoid organs, conferred partial protection against subcutaneous tumors, and prevented the establishment of lung metastasis. Taken together, our data support the use of cell-free, synthetic aAPC as a specific and versatile alternative to expand peptide-specific CD4(+) T cells in adoptive and active immunotherapy. PMID- 18413772 TI - Interleukin 21 enhances antibody-mediated tumor rejection. AB - Interleukin-21 (IL-21) is a cytokine with structural and sequence homology to IL 2 and IL-15 that has antitumor activity alone in mouse experimental tumor models and a tolerable safety profile in phase I trials in patients with metastatic melanoma and renal cell carcinoma. Several monoclonal antibodies (mAb) targeted at tumor-associated antigens also have improved antitumor activities in mice when used in combination with IL-21. Recently, we described a rational three antibody based approach (triple mAb, TrimAb) to eradicating established mouse tumors that required the generation of tumor-reactive CD8(+) T cells and IFN-gamma. Herein, we show that sequentially combining TrimAb with recombinant IL-21 can significantly improve the antitumor activity of this combination against very advanced disease. These data further support the use of IL-21 in adjuvant settings where strong T cell-mediated immune responses to tumors can be generated. PMID- 18413773 TI - The helix-loop-helix protein Id1 requires cyclin D1 to promote the proliferation of mammary epithelial cell acini. AB - Overexpression of the helix-loop-helix (HLH) protein Id1 has been associated with metastasis in breast cancer, but its role in models of early breast tumorigenesis is not well characterized. We show that the down-regulation of endogenous Id1 via proteosomal degradation and relocalization from the nucleus to the cytoplasm is an early event in the formation of mammary epithelial acini. Overexpression of Id1 in both human MCF-10A and primary mouse mammary epithelial cells disrupted normal acinar development by increasing acinar volume. This occurred in an HLH domain-dependent fashion via an increase in S phase. Id1 overexpression also increased apoptosis leading to accelerated luminal clearance, and this was reversed by coexpression of the proto-oncogene Bcl2, leading to large, disorganized structures with filled lumina. Id1 overexpression was unable to increase the volume of cyclin D1(-/-) acini, indicating that Id1 is dependent on cyclin D1 for its proliferative effects. In summary, Id1 may contribute to early breast cancer by promoting excessive proliferation through cyclin D1. PMID- 18413774 TI - Gender-related survival differences associated with EGFR polymorphisms in metastatic colon cancer. AB - Evidence is accumulating supporting gender-related differences in the development of colonic carcinomas. Sex steroid hormone receptors are expressed in the colon and interact with epidermal growth factor receptor (EGFR), a gene widely expressed in colonic tissue. Increased EGFR expression is linked with poor prognosis in colon cancer. Within the EGFR gene there are two functional polymorphisms of interest: a polymorphism located at codon 497 (HER-1 R497K) and a dinucleotide (CA)(n) repeat polymorphism located within intron 1. These germ line polymorphisms of EGFR were analyzed in genomic DNA from 318 metastatic colon cancer patients, 177 males and 141 females, collected from 1992 to 2003. Gender related survival differences were associated with the HER-1 R497K polymorphism (P(interaction) = 0.003). Females with the HER-1 497 Arg/Arg variant had better overall survival (OS) when compared with the Lys/Lys and/or Lys/Arg variants. In males the opposite was true. The EGFR dinucleotide (CA)(n) repeat also trended with a gender-related OS difference (P(interaction) = 0.11). Females with both short <20 (CA)(n) repeat alleles had better OS than those with any long >or=20 (CA)(n) repeats. In males the opposite was true. Combination analysis of the two polymorphisms taken together also revealed the same gender-related survival difference (P(interaction) = 0.002). These associations were observed using multivariable analysis. The two polymorphisms were not in linkage disequilibrium and are independent of one another. This study supports the role of functional EGFR polymorphisms as independent prognostic markers in metastatic colon cancer. As a prognostic factor, these variants had opposite prognostic implications based on gender. PMID- 18413775 TI - Genetic variation in the inhibin pathway and risk of testicular germ cell tumors. AB - Gene-knockout studies in mice suggest that INHA, encoding a subunit of gonadotropin-regulating proteins known as inhibins, is a tumor suppressor for testicular stromal cell tumors. It is not known whether genetic variation in the inhibin pathway also influences susceptibility to testicular germ cell tumors (TGCT), the most common testicular cancer in young men. To address this question, we conducted a case-control analysis (577 cases; 707 controls) of single nucleotide polymorphisms (SNP) in genes in the inhibin pathway among participants in the U.S. Servicemen's Testicular Tumor Environmental and Endocrine Determinants Study. Thirty-eight tagging SNPs in six genes (INHA, INHBA, INHBB, INHBC, INHBE, and SMAD4) were genotyped. Odds ratios (OR) and 95% confidence intervals (CI) relating variant genotypes to TGCT risk were calculated using unconditional logistic regression. Among White subjects, an elevated risk of TGCT was observed for carriers of the T allele of the INHA variant rs2059693 (CT genotype: OR, 1.33; 95% CI, 1.04-1.71; TT: OR, 1.60; 95% CI, 1.01-2.52; P(trend) = 0.008). The association with rs2059693 was stronger for nonseminomas, and for teratomas and teratocarcinomas in particular (N = 58; CT: OR, 1.63; 95% CI, 0.89 2.99; TT: OR, 4.54; 95% CI 2.00-10.3; P(trend) = 0.0008). We found no evidence of association with variants in the other investigated genes. These findings suggest that genetic variation in the INHA locus influences TGCT development. PMID- 18413776 TI - Double-strand break damage and associated DNA repair genes predispose smokers to gene methylation. AB - Gene promoter hypermethylation in sputum is a promising biomarker for predicting lung cancer. Identifying factors that predispose smokers to methylation of multiple gene promoters in the lung could affect strategies for early detection and chemoprevention. This study evaluated the hypothesis that double-strand break (DSB) repair capacity and sequence variation in genes in this pathway are associated with a high methylation index in a cohort of current and former cancer free smokers. A 50% reduction in the mean level of DSB repair capacity was seen in lymphocytes from smokers with a high methylation index, defined as three or more of eight genes methylated in sputum, compared with smokers with no genes methylated. The classification accuracy for predicting risk for methylation was 88%. Single nucleotide polymorphisms within the MRE11A, CHEK2, XRCC3, DNA-PKc, and NBN DNA repair genes were highly associated with the methylation index. A 14.5-fold increased odds for high methylation was seen for persons with seven or more risk alleles of these genes. Promoter activity of the MRE11A gene that plays a critical role in recognition of DNA damage and activation of ataxia telangiectasia mutated was reduced in persons with the risk allele. Collectively, ours is the first population-based study to identify DSB DNA repair capacity and specific genes within this pathway as critical determinants for gene methylation in sputum, which is, in turn, associated with elevated risk for lung cancer. PMID- 18413777 TI - Enhancement of UVB-induced apoptosis by apigenin in human keratinocytes and organotypic keratinocyte cultures. AB - Topical application of the bioflavonoid 4',5,7-trihydroxyflavone (apigenin) to mouse skin effectively reduces the incidence and size of skin tumors caused by UVB exposure. The ability to act as a chemopreventive compound indicates that apigenin treatment alters the molecular events initiated by UVB exposure; however, the effects of apigenin treatment on UVB-irradiated keratinocytes are not fully understood. In the present study, we have used three models of human keratinocytes to study the effect of apigenin treatment on UVB-induced apoptosis: HaCaT human keratinocyte cells, primary keratinocyte cultures isolated from human neonatal foreskin, and human organotypic keratinocyte cultures. Each keratinocyte model was exposed to a moderate dose of UVB (300-1,000 J/m(2)), then treated with apigenin (0-50 micromol/L), and harvested to assess apoptosis by Western blot analysis for poly(ADP)ribose polymerase cleavage, annexin-V staining by flow cytometry, and/or the presence of sunburn cells. Apigenin treatment enhanced UVB induced apoptosis >2-fold in each of the models tested. When keratinocytes were exposed to UVB, apigenin treatment stimulated changes in Bax localization and increased the release of cytochrome c from the mitochondria compared with UVB exposure alone. Overexpression of the antiapoptotic protein Bcl-2 and expression of a dominant-negative form of Fas-associated death domain led to a reduction in the ability of apigenin to enhance UVB-induced apoptosis. These results suggest that enhancement of UVB-induced apoptosis by apigenin treatment involves both the intrinsic and extrinsic apoptotic pathways. The ability of apigenin to enhance UVB-induced apoptosis may explain, in part, the photochemopreventive effects of apigenin. PMID- 18413779 TI - A statistical reappraisal of the findings of an esophageal cancer genome-wide association study. PMID- 18413778 TI - Effect of low-fat diet on development of prostate cancer and Akt phosphorylation in the Hi-Myc transgenic mouse model. AB - This study evaluated the effect of dietary fat on prostate cancer development by using the Hi-Myc mouse transgenic prostate cancer model. Hi-Myc mice develop murine prostatic intraepithelial neoplasia (mPIN) as early as 2 to 4 weeks and invasive adenocarcinoma between 6 and 9 months due to the overexpression of human c-Myc in the mouse prostate. Three-week-old male Hi-Myc mice were placed on high fat (HF; 42% Kcal) or low-fat (LF; 12% Kcal) diets, and equal caloric intake was maintained until euthanasia at 7 months. The number of mice that developed invasive adenocarcinoma at 7 months was 27% less in the LF diet group (12/28) compared with the HF diet group (23/33, P < 0.05). Epithelial cells in mPIN lesions in the LF group had a significantly lower proliferative index compared with epithelial cells in the HF group (21.7% versus 28.9%, P < 0.05). During the mPIN phase of carcinogenesis (4 months), the LF group had higher serum insulin like growth factor (IGF) binding protein-1 levels (21.0 +/- 8.9 ng/mL versus 3.2 +/- 0.8 ng/mL, P < 0.05) relative to the HF group. Akt (Ser(473)) phosphorylation, Akt kinase activity, and phosphorylation of downstream targets of Akt in prostates were significantly reduced in the LF diet group compared with the HF group. We conclude that dietary fat reduction delays transition from mPIN to invasive cancer in this Myc-driven transgenic mouse model, possibly through suppression of the IGF-Akt pathway and decreased proliferation of mPIN epithelial cells. PMID- 18413781 TI - Overlapping genes in Nalp6/PYPAF5 locus encode two V2-type vasopressin isoreceptors: angiotensin-vasopressin receptor (AVR) and non-AVR. AB - The angiotensin-vasopressin receptor (AVR) responds with equivalent affinities to angiotensin II (ANG II) and vasopressin and is coupled to adenylate cyclase and hence a V2-type vasopressin receptor. AVR maps to the Nalp6 locus and overlaps with the larger Nalp6/PYPAF5 reported to be a T cell/granulocyte-specific, cytoplasmic-specific proapoptotic protein, thus questioning the existence of AVR. Here we confirm, through different experimental modalities, that AVR is distinct from Nalp6/PYPAF5 based on different mRNA and protein sizes, subcellular localization, and tissue-specific expression patterns. Binding studies of PYPAF5 specific Cos1 transfectants detect high-affinity binding to vasopressin but not ANG II, thus assigning PYPAF5 as a non-AVR (NAVR). Signaling array analysis reveals that AVP stimulation of AVR- and NAVR-specific Cos1 transfectants results in diametrical activation as well as coactivation of signaling pathways known to mediate renal sodium and water balance. Likewise, ANG II stimulation of Cos1-AVR transfectants reveals a signaling profile distinct from that of AVP-stimulated Cos1-AVR transfectants. Analysis of genomic organization of the AVR/NAVR locus shows an overlapping gene arrangement with alternative promoter usage resulting in different NH(2) termini for NAVR and AVR. In addition to core promoter elements, androgen and estrogen response elements are detected. Promoter analysis of NAVR/AVR 5'-regulatory region detects transcriptional upregulation by testosterone and synergistic upregulation by testosterone and estrogen, thus suggesting that AVR and/or NAVR contribute to sex-specific V2-type vasopressin mediated effects. Altogether, confirmation of AVR and identification of NAVR as vasopressin receptors are concordant with emerging vasopressin functions not attributable to V1a, V1b, or V2 receptors and add molecular bases for the multifunctional complexity of vasopressin-mediated functions and regulation. PMID- 18413782 TI - Current challenges in metabolomics for diabetes research: a vital functional genomic tool or just a ploy for gaining funding? AB - Metabolomics aims to profile all the small molecule metabolites found within a cell, tissue, organ, or organism and use this information to understand a biological manipulation such as a drug intervention or a gene knockout. While neither mass spectrometry or NMR spectroscopy, the two most commonly used analytical tools in metabolomics, can provide a complete coverage of the metabolome, compared with other functional genomic tools for profiling biological moieties the approach is cheap and high throughput. In diabetes and obesity research this has provided the opportunity to assess large human populations or investigate a range of different tissues in animal studies both rapidly and cheaply. However, the approach has a number of major challenges, particularly with the interpretation of the data obtained. For example, some key pathways are better represented by high concentration metabolites inside the cell, and thus, the coverage of the metabolome may become biased towards these pathways (e.g., the TCA cycle, amino acid metabolism). There is also the challenge of statistically modeling datasets with large numbers of variables but relatively small sample sizes. This perspective discusses our own experience of some of the benefits and pitfalls with using metabolomics to understand diseases associated with type 2 diabetes. PMID- 18413783 TI - Processing of naturally occurring sense/antisense transcripts of the vertebrate Slc34a gene into short RNAs. AB - Overlapping sense/antisense RNAs transcribed in opposite directions from the same genomic locus are common in vertebrates. The impact of antisense transcription on gene regulation and cell biology is largely unknown. We show that sense/antisense RNAs of an evolutionarily conserved phosphate transporter gene (Slc34a2a) are coexpressed in a short time window during embryonic development of zebrafish at 48 hours postfertilization (hpf). To address the mechanism by which coexpressed sense/antisense transcripts are processed, we injected sense/antisense RNAs in various combinations into Xenopus oocytes. In the cytoplasm RNAs were stable in whatever combination expressed. In the nucleus coinjected sense/antisense transcripts were degraded into short RNAs of approximately 23 bases within 4 h. A homologous transcript from toad or another isoform (Slc34a2b) from zebrafish failed to trigger processing. In oocytes that were primed with nuclear sense/antisense RNA coinjections, a reporter RNA was rapidly degraded. We produced evidence that the observed processing of complementary transcripts was not restricted to Xenopus oocytes, because Slc34a-related short RNAs were detected in zebrafish embryos by Northern blotting. Signals were observed at stages that showed coexpression of sense/antisense transcripts. Remarkably, strand-specific probes revealed that the orientation of short RNAs was developmentally regulated. In addition, RNA from zebrafish embryos 48 hpf was able to induce degradation of reporter constructs in Xenopus oocytes. Our findings may give important clues to understanding the physiological role of the widespread antisense transcription. PMID- 18413785 TI - Response to letter of Vahle et al. PMID- 18413784 TI - Potassium channels: newly found players in synaptic plasticity. AB - One of the major issues for modern neuroscience research concerns the molecular and cellular mechanisms that underlie the acquisition, storage, and recollection of memories by the brain. Regulation of the strength of individual synaptic inputs (synaptic plasticity) has, for decades, been the front-running candidate mechanism for cellular information storage, with some direct supporting evidence recently obtained. Research into the molecular mechanisms responsible for changing synaptic strength has, to date, primarily focused on trafficking and properties of the neurotransmitter receptors themselves (AMPARs and NMDARs). However, recent evidence indicates that, subsequent to receptor activation, synaptic inputs are subject to regulation by synaptically located K+ channels. It is therefore critical to understand the biophysical properties and subcellular localization (density and distribution) of these channels and how their properties are modulated. Here we will review recent findings showing that two different classes of K+ channels (A-type and small conductance, Ca2+ -activated K+ channels), beyond their traditional role in regulating action potential firing, contribute to the regulation of synaptic strength in the hippocampus. In addition, we discuss how modulation of these channels' properties and expression might contribute to synaptic plasticity. PMID- 18413786 TI - Temporal gene expression profiling indicates early up-regulation of interleukin-6 in isoproterenol-induced myocardial necrosis in rat. AB - Gene expression was evaluated in the myocardium of male Wistar rats after a single subcutaneous administration of 0.5 mg of isoproterenol, a beta-adrenergic agonist that causes acute tachycardia with subsequent myocardial necrosis. Histology of the heart, clinical chemistry, and hematology were evaluated at 9 time points (0.5 hours to 14 days postinjection). Myocardial gene expression was evaluated at 4 time points (1 hour to 3 days). Contraction bands and loss of cross-striation were identified on phosphotungstic acid-hematoxylin-stained sections 0.5 hours postdosing. Plasma troponin I elevation was detected at 0.5 hours, peaked at 3 hours, and returned to baseline values at 3 days postdosing. Interleukin 6 (Il6) expression spiked at 1 to 3 hours and was followed by a short lived, time-dependent dysregulation of its downstream targets. Concurrently and consistent with the kinetics of the histologic findings, many pathways indicative of necrosis/apoptosis (p38 mitogen-activated protein kinase [MAPK] signaling, NF kappaB signaling) and adaptation to hypertension (PPAR signaling) were overrepresented at 3 hours. The 1-day and 3-day time points indicated an adaptive response, with down-regulation of the fatty acid metabolism pathway, up regulation of the fetal gene program, and superimposed inflammation and repair at 3 days. These results suggest early involvement of Il6 in isoproterenol-induced myocardial necrosis and emphasize the value of early time points in transcriptomic studies. PMID- 18413787 TI - Neuroexcitatory targets in the female reproductive system of the nonhuman primate (Macaca fascicularis). AB - Glutamate receptors (GluRs) have been implicated in brain function and pathology. Their presence in peripheral tissues suggests a vital role in the pathophysiology of various organ systems. In earlier studies, the authors reported the differential distribution of ionotropic and metabotropic GluRs in neural and nonneural peripheral tissues of the rat. In this study, they investigated the presence and the localization of the GluRs in the reproductive organs of Macaca fascicularis. The data illustrate the presence of the GluR 2/3, metabotropic glutamate receptor 2/3, kainate 2, and N-methyl-D-aspartate receptor 1 (NMDAR 1). These are localized in the different structures of the ovaries, uterine cervix, myometrium, endometrium, and inflammatory cells. Smooth muscle of the myometrium and arterioles showed strong immunolabeling with anti-GluR 2/3 and, to a lesser intensity, with the other ionotropic glutamate receptor antibodies. NMDAR 1 showed the most widespread staining in all the structures. Mast cells showed strong immunolabeling with the anti-NMDA antibody. The demonstration and the differential expression of GluRs in the female reproductive system of nonhuman primate experimental models provide first evidence suggesting excitatory signaling in these tissues. PMID- 18413788 TI - Evidence of a threshold-effect for 2-amino-3,8-dimethylimidazo-[4,5-f]quinoxaline liver carcinogenicity in F344/DuCrj rats. AB - To estimate potential human risk of exposure to a food-derived, genotoxic hepatocarcinogen, 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx), a 2-year carcinogenicity test was conducted using male F344 rats administered MeIQx containing diet at doses of 0 (control), 0.001, 1, and 100 ppm. The lowest dose 0.001 ppm was established as equivalent to the daily intake of this carcinogen in humans (0.2 to 2.6 microg/man/day). Significant decreases of survival rate and body weight gain were observed in rats treated with 100 ppm MeIQx. Histopathological examination revealed significant induction of hepatocellular carcinomas, adenomas, and development of glutathione S-transferase placental form positive foci with MeIQx at 100 ppm. Moreover, the incidences of Zymbal's glands carcinoma, mammary fibroadenoma, and subcutaneous fibroma were found significantly increased in a 100 ppm MeIQx group. However, no significant induction of altered preneoplastic hepatocellular foci was observed in 0.001 and 1 ppm groups as compared to the controls. 8-Hydroxy-2'-deoxyguanosine levels in the rat liver DNA of the 100 ppm-treated group were not elevated, but MeIQx-DNA adduct formation increased as compared with the 1 ppm case, albeit without significance. No significant induction of any other neoplastic lesions related to the carcinogen administration was found in MeIQx-administered groups except for 100 ppm. These results imply that 1 ppm may be a no-effect level for MeIQx carcinogenesis. PMID- 18413789 TI - Histone tail modifications and noncanonical functions of histones: perspectives in cancer epigenetics. AB - Over the past few years, the histone deacetylase (HDAC) inhibitors have occupied an important place in the effort to develop novel, but less toxic, anticancer therapy. HDAC inhibitors block HDACs, which are the enzymes responsible for histone deacetylation, and therefore they modulate gene expression. The cellular effects of HDAC inhibitors include growth arrest and the induction of differentiation. Early successes in cancer therapeutics obtained using these drugs alone or in combination with other anticancer drugs emphasize the important place of posttranslational modifications of histones in cancer therapy. Histone tail modifications along with DNA methylation are the most studied epigenetic events related to cancer progression. Moreover, extranuclear functions of histones have also been described. Because HDAC inhibitors block HDACs and thereby increase histone acetylation, we propose a model wherein exogenous acetylated histones or other related acetylated proteins that are introduced into the nucleus become HDAC substrates and thereby compete with endogenous histones for HDACs. This competition may lead to the increased acetylation of the endogenous histones, as in the case of HDAC inhibitor therapy. Moreover, other mechanisms of action, such as binding to chromatin and modulating gene expression, are also possible for exogenously introduced histones. PMID- 18413790 TI - MGCD0103, a novel isotype-selective histone deacetylase inhibitor, has broad spectrum antitumor activity in vitro and in vivo. AB - Nonselective inhibitors of human histone deacetylases (HDAC) are known to have antitumor activity in mice in vivo, and several of them are under clinical investigation. The first of these, Vorinostat (SAHA), has been approved for treatment of cutaneous T-cell lymphoma. Questions remain concerning which HDAC isotype(s) are the best to target for anticancer activity and whether increased efficacy and safety will result with an isotype-selective HDAC inhibitor. We have developed an isotype-selective HDAC inhibitor, MGCD0103, which potently targets human HDAC1 but also has inhibitory activity against HDAC2, HDAC3, and HDAC11 in vitro. In intact cells, MGCD0103 inhibited only a fraction of the total HDAC activity and showed long-lasting inhibitory activity even upon drug removal. MGCD0103 induced hyperacetylation of histones, selectively induced apoptosis, and caused cell cycle blockade in various human cancer cell lines in a dose-dependent manner. MGCD0103 exhibited potent and selective antiproliferative activities against a broad spectrum of human cancer cell lines in vitro, and HDAC inhibitory activity was required for these effects. In vivo, MGCD0103 significantly inhibited growth of human tumor xenografts in nude mice in a dose-dependent manner and the antitumor activity correlated with induction of histone acetylation in tumors. Our findings suggest that the isotype-selective HDAC inhibition by MGCD0103 is sufficient for antitumor activity in vivo and that further clinical investigation is warranted. PMID- 18413791 TI - Small molecules targeting histone H4 as potential therapeutics for chronic myelogenous leukemia. AB - We recently identified a polyamide-chlorambucil conjugate, 1R-Chl, which alkylates and down-regulates transcription of the human histone H4c gene and inhibits the growth of several cancer cell lines in vitro and in a murine SW620 xenograft model, without apparent animal toxicity. In this study, we analyzed the effects of 1R-Chl in the chronic myelogenous leukemia cell line K562 and identified another polyamide conjugate, 6R-Chl, which targets H4 genes and elicits a similar cellular response. Other polyamide conjugates that do not target the H4 gene do not elicit this response. In a murine model, both 1R-Chl and 6R-Chl were found to be highly effective in blocking K562 xenograft growth with high-dose tolerance. Unlike conventional and distamycin-based alkylators, little or no cytotoxicities and animal toxicities were observed in mg/kg dosage ranges. These results suggest that these polyamide alkylators may be a viable treatment alternative for chronic myelogenous leukemia. PMID- 18413792 TI - Histone deacetylase inhibitor FK228 enhances adenovirus-mediated p53 family gene therapy in cancer models. AB - Therapeutic replacement of the wild-type p53 gene has been pursued as a potential gene therapy strategy in a variety of cancer types; however, some cancer models are resistant to p53 in vivo and in vitro. Therefore, to improve p53 gene therapy, it is important to overcome the resistance to p53-mediated apoptosis. Histone deacetylase inhibitors are a novel class of chemotherapeutic agents that are able to reverse the malignant phenotype of transformed cells. A natural histone deacetylase inhibitor, FK228, is reported to enhance adenovirus infection due in part to the up-regulation of coxsackievirus adenovirus receptor expression. In this study, preclinical experiments were done to establish a mechanistic rationale for the combination of adenovirus-mediated p53 family gene transfer and FK228 pretreatment in future clinical trials. Pretreatment with FK228 enhanced apoptosis in human cancer cells through enhanced transduction of Ad-p53. FK228 also induced hyperacetylation of the p53 protein and specifically enhanced p53-mediated Noxa expression. Additionally, the combination of FK228 and Ad-p53 induced Bax translocation to the mitochondria. The double knockdown of Bax and Noxa expression by small interfering RNA antagonized the synergistic effect of Ad-p53 and FK228 on apoptosis induction. In human cancer xenograft models, FK228 significantly increased the therapeutic effectiveness of p53 as well as p63 gene therapy. These results provide a strong rationale for combining p53 gene therapy and FK228 pretreatment in cancer therapy. PMID- 18413793 TI - Sensitizing estrogen receptor-negative breast cancer cells to tamoxifen with OSU 03012, a novel celecoxib-derived phosphoinositide-dependent protein kinase-1/Akt signaling inhibitor. AB - Tamoxifen is a mainstay in the treatment of estrogen receptor (ER)-positive breast cancer patients. Although the efficacy of tamoxifen has been attributed to induction of tumor cell growth arrest and apoptosis by inhibition of ER signaling, recent evidence indicates that tamoxifen possesses ER-independent antitumor activities. Here, we use OSU-03012, a small-molecule inhibitor of phosphoinositide-dependent protein kinase-1 (PDK-1) to address the hypothesis that PDK-1/Akt signaling represents a therapeutically relevant target to sensitize ER-negative breast cancer to tamoxifen. OSU-03012 sensitized both ER positive MCF-7 and ER-negative MDA-MB-231 cells to the antiproliferative effects of tamoxifen in an ER-independent manner. Flow cytometric analysis of phosphatidylserine externalization revealed that this augmented suppression of cell viability was attributable to a marked enhancement of tamoxifen-induced apoptosis by OSU-03012. Mechanistically, this OSU-03012-mediated sensitization was associated with suppression of a transient tamoxifen-induced elevation of Akt phosphorylation and enhanced modulation of the functional status of multiple Akt downstream effectors, including FOXO3a, GSK3alpha/beta, and p27. The growth of established MDA-MB-231 tumor xenografts was suppressed by 50% after oral treatment with the combination of tamoxifen (60 mg/kg) and OSU-03012 (100 mg/kg), whereas OSU-03012 and tamoxifen alone suppressed growth by 30% and 0%, respectively. These findings indicate that the inhibition of PDK-1/Akt signaling to sensitize ER-negative breast cancer cells to the ER-independent antitumor activities of tamoxifen represents a feasible approach to extending the use of tamoxifen to a broader population of breast cancer patients. Considering the urgent need for novel therapeutic strategies for ER-negative breast cancer patients, this combinatorial approach is worthy of continued investigation. PMID- 18413794 TI - 2-Deoxyglucose induces Akt phosphorylation via a mechanism independent of LKB1/AMP-activated protein kinase signaling activation or glycolysis inhibition. AB - The compound 2-deoxyglucose (2-DG) enhances chemotherapy/radiotherapy in cell lines and animal models, prompting two phase I clinical trials with this cancer therapeutic. Although its mechanism of action has not been fully elucidated, it is hypothesized that the molecular basis of 2-DG activity is related to glycolysis inhibition. Here, we report that 2-DG induced Akt phosphorylation at Thr(308) and Ser(473) as early as 15 min post-treatment. These phosphorylation events required phosphatidylinositol-3-kinase activity but were not related to LKB1/AMP-activated protein kinase signaling, the inhibition of glycolysis or epidermal growth factor receptor signaling. The 2-DG-mediated Akt phosphorylation also led to the phosphorylation of Akt downstream targets, such as Foxo3a, GSK3beta, and Chk1. Because the functional consequence of Akt activation includes chemotherapy/radiotherapy resistance, our data suggested that the combination of phosphatidylinositol-3-kinase/Akt inhibitory agents in 2-DG-based chemotherapy/radiotherapy may result in enhanced therapeutic efficacy. PMID- 18413795 TI - Pharmacologic properties of AG-012986, a pan-cyclin-dependent kinase inhibitor with antitumor efficacy. AB - AG-012986 is a multitargeted cyclin-dependent kinase (CDK) inhibitor active against CDK1, CDK2, CDK4/6, CDK5, and CDK9, with selectivity over a diverse panel of non-CDK kinases. Here, we report the potent antitumor efficacies of AG-012986 against multiple tumor lines in vitro and in vivo. AG-012986 showed antiproliferative activities in vitro with IC(50)s of <100 nmol/L in 14 of 18 tumor cell lines. In vivo, significant antitumor efficacy induced by AG-012986 was seen (tumor growth inhibition, >83.1%) in 10 of 11 human xenograft tumor models when administered at or near the maximum tolerated dose for 8 or 12 days. AG-012986 caused dose-dependent hypophosphorylation at Ser(795) of the retinoblastoma protein, cell cycle arrest, and apoptosis in vitro. Colony-forming assays indicated that the potency of AG-012986 substantially decreased with treatment time of <24 h. In vivo, AG-012986 also showed dose-dependent retinoblastoma Ser(795) hypophosphorylation, cell cycle arrest, decreased Ki-67 tumor staining, and apoptosis in conjunction with antitumor activity. Studies comparing i.p. bolus with s.c. implanted minipump dosing regimens revealed that in vivo efficacy correlated with the duration of minimally effective plasma levels rather than maximal drug plasma levels. Dosing optimization of AG-012986 provided guidance for selecting a treatment schedule to achieve the best antitumor efficacy while minimizing the risk of adverse side effects. PMID- 18413796 TI - LY2109761, a novel transforming growth factor beta receptor type I and type II dual inhibitor, as a therapeutic approach to suppressing pancreatic cancer metastasis. AB - Most pancreatic cancer patients present with inoperable disease or develop metastases after surgery. Conventional therapies are usually ineffective in treating metastatic disease. It is evident that novel therapies remain to be developed. Transforming growth factor beta (TGF-beta) plays a key role in cancer metastasis, signaling through the TGF-beta type I/II receptors (TbetaRI/II). We hypothesized that targeting TbetaRI/II kinase activity with the novel inhibitor LY2109761 would suppress pancreatic cancer metastatic processes. The effect of LY2109761 has been evaluated on soft agar growth, migration, invasion using a fibroblast coculture model, and detachment-induced apoptosis (anoikis) by Annexin V flow cytometric analysis. The efficacy of LY2109761 on tumor growth, survival, and reduction of spontaneous metastasis have been evaluated in an orthotopic murine model of metastatic pancreatic cancer expressing both luciferase and green fluorescence proteins (L3.6pl/GLT). To determine whether pancreatic cancer cells or the cells in the liver microenvironment were involved in LY2109761-mediated reduction of liver metastasis, we used a model of experimental liver metastasis. LY2109761 significantly inhibited the L3.6pl/GLT soft agar growth, suppressed both basal and TGF-beta1-induced cell migration and invasion, and induced anoikis. In vivo, LY2109761, in combination with gemcitabine, significantly reduced the tumor burden, prolonged survival, and reduced spontaneous abdominal metastases. Results from the experimental liver metastasis models indicate an important role for targeting TbetaRI/II kinase activity on tumor and liver microenvironment cells in suppressing liver metastasis. Targeting TbetaRI/II kinase activity on pancreatic cancer cells or the cells of the liver microenvironment represents a novel therapeutic approach to prevent pancreatic cancer metastasis. PMID- 18413797 TI - Characterization of structurally distinct, isoform-selective phosphoinositide 3' kinase inhibitors in combination with radiation in the treatment of glioblastoma. AB - The phosphoinositide 3'-kinase (PI3K)-mediated signaling pathway plays a key role in fundamental cellular functions important in normal cellular homeostasis and malignant transformation. Deregulated signaling through this pathway contributes to development of gliomas and their resistance to radiation and chemotherapy. Targeting the PI3K signaling pathway has thus emerged as a promising approach to successful treatment of gliomas. We assessed the radiosensitizing potential of four small-molecule inhibitors that differ in their activities against specific isoforms of the PI3K 110-kDa catalytic subunit (p110). p110alpha inhibitors blocked phosphorylation of both protein kinase B/Akt and S6 in all cell lines examined, effectively decreased cellular proliferation, and produced additive cytotoxic effects in combination with radiation therapy. The p110beta inhibitor exhibited limited biochemical effects and failed to decrease cellular proliferation or viability as either a single agent or in combination with radiation or rapamycin. In vivo studies examining the effects of the p110alpha inhibitor in combination with radiation indicated a significant reduction in tumor growth rate induced by the combined treatment compared with each treatment modality alone. This translated into a trend toward prolonged time-to-failure for mice in the combination treatment group. In conclusion, PI3K inhibitors are promising agents in the treatment of glioblastomas, especially when used in combination with ionizing radiation. PMID- 18413798 TI - Enhancement of antitumor properties of TRAIL by targeted delivery to the tumor neovasculature. AB - Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a promising anticancer agent with tumor-selective apoptotic activity. TRAIL plays a role in the innate and adaptive immune response and autoimmune disease and may also be involved in hepatic cell death and inflammation. For these reasons, chronic exposure to TRAIL may have deleterious side effects in patients as a cancer therapeutic. In this study, we have improved the antitumor activity of TRAIL by targeted delivery to the tumor vasculature, leading to dramatic enhancement of its therapeutic properties. TRAIL was fused to the ACDCRGDCFC peptide (named RGD L-TRAIL), a ligand of alpha(V)beta(3) and alpha(V)beta(5) integrins. Biological activity was evaluated in vitro and antitumor efficacy was investigated in vivo as a single agent and in combination with irinotecan hydrochloride (CPT-11). The fusion protein RGD-L-TRAIL, but not TRAIL or RGE-L-TRAIL, specifically bound to microvascular endothelial cells in a dose-dependent manner and showed enhanced apoptosis-inducing activity (caspase-3 and caspase-8 activation) in alpha(V)beta(3) and alpha(V)beta(5) integrin-positive cancer cells. In addition, RGD-L-TRAIL was more effective in suppressing tumor growth of COLO-205 tumor bearing mice than an equivalent dose of TRAIL. The antitumor effect of RGD-L TRAIL was further enhanced by combination with CPT-11 in both TRAIL-sensitive COLO-205 and TRAIL-resistive HT-29 tumor xenograft models. Our findings suggest that the novel fusion protein RGD-L-TRAIL can directly target tumor endothelial cells as well as alpha(V)beta(3) and alpha(V)beta(5) integrin-positive tumor cells. The tumor-targeted delivery of TRAIL derivatives, such as RGD-L-TRAIL, may prove to be a promising lead candidate for cancer therapy. PMID- 18413799 TI - Antitumor activity of fibroblast growth factor receptor 3-specific immunotoxins in a xenograft mouse model of bladder carcinoma is mediated by apoptosis. AB - Human single-chain Fv directed against fibroblast growth factor receptor 3 (FGFR3) have been shown to block proliferation of RT112 bladder carcinoma cells in vitro. Here, we examined the ability of the recombinant gelonin toxin (rGel) to enhance this inhibitory effect in vitro and in vivo on the bladder cancer cell line RT112 and the corresponding xenografts. Immunotoxins were genetically engineered by fusing FGFR3-specific Fv fragments (3C) to the NH(2) terminus of rGel and expressed as a soluble protein in Escherichia coli. The 3C/rGel fusion construct showed an IC(50) of 200 nmol/L against log-phase RT112 cells compared with 1,500 nmol/L for free rGel. Immunofluorescence studies showed that the 3C/rGel construct internalized rapidly into the cytoplasm of RT112 cells within 1 h of exposure. The mechanism of immunotoxin-induced cell death was found to be mediated by apoptosis. RT112 tumor xenografts in severe combined immunodeficient mice treated with 50 mg/kg 3C/rGel exhibited considerable growth delay relative to control tumors and a significant reduction of 55% to 70% in mean tumor size. Immunohistochemical analysis showed that tumors from mice treated with 3C/rGel displayed considerable apoptotic damage compared with control groups. Subcellular location of FGFR3 in immunotoxin-treated tumors indicated a translocation of FGFR3 to the nuclear membrane in contrast to tumors from saline-treated controls. These results show that FGFR3-driven immunotoxins may be an effective therapeutic agent against human bladder and other tumor types overexpressing FGFR3. PMID- 18413800 TI - The T790M "gatekeeper" mutation in EGFR mediates resistance to low concentrations of an irreversible EGFR inhibitor. AB - Patients with non-small cell lung cancer (NSCLC) harboring activating mutations in the epidermal growth factor receptor (EGFR) kinase domain tend to respond well to the tyrosine kinase inhibitors, gefitinib and erlotinib. However, following clinical response, these patients typically relapse within a year of treatment. In many cases, resistance is caused by an acquired secondary EGFR kinase domain mutation, T790M. In vitro studies have shown that a new class of EGFR irreversible inhibitors could overcome the resistance conferred by T790M. Clinical trials are under way to examine the efficacy of one of these inhibitors, HKI-272, in patients with NSCLC who initially responded to gefitinib/erlotinib and subsequently relapsed. To anticipate the possibility that patients who respond to irreversible inhibitors will develop secondary resistance to such inhibitors, as has been seen in other similar settings, we modeled acquired resistance to the dual EGFR/HER2-irreversible tyrosine kinase inhibitor HKI-272 in a NSCLC cell culture model. We found that HKI-272-resistant clones fall into two biochemical groups based on the retention of EGFR phosphorylation in the presence of the drug. Cells that retain phosphorylated EGFR have acquired the secondary mutation T790M. Moreover, HKI-272 can overcome T790M resistance only at suprapharmacologic concentrations. We further model mutations at EGFR C797 as a mechanism of resistance to irreversible EGFR inhibitors and show that although these mutants are resistant to the irreversible inhibitor, they retain erlotinib sensitivity. Our findings suggest that HKI-272 treatment at maximally tolerated dosing may lead to the emergence of T790M-mediated resistance, whereas treatment with a more potent irreversible inhibitor could yield a resistance mutation at EGFR C797. PMID- 18413802 TI - Sorafenib inhibits growth and mitogen-activated protein kinase signaling in malignant peripheral nerve sheath cells. AB - Malignant peripheral nerve sheath tumors (MPNST) are soft-tissue tumors with a very poor prognosis and largely resistant to chemotherapy. MPNSTs are characterized by activation of the Ras pathway by loss of tumor suppressor neurofibromatosis type 1. In view of this, MPNST may be susceptible to inhibition of the activated Ras/Raf/mitogen-activated protein kinase pathway by the B-Raf inhibitor sorafenib. MPNST (MPNST and ST8814) and dedifferentiated liposarcoma (LS141 and DDLS) human tumor cell lines were characterized for Ras activation and B-Raf expression. Tumor cells were treated with sorafenib and examined for growth inhibition, inhibition of phospho-MEK, phospho-ERK, cell cycle arrest, and changes in cyclin D1 and pRb expression. MPNSTs were sensitive to sorafenib at nanomolar concentrations. This appeared to be due to inhibition of phospho-MEK, phospho-ERK, suppression of cyclin D1, and hypophosphorylation of pRb at the CDK4 specific sites, resulting in a G(1) cell cycle arrest. These effects were not seen in the liposarcoma cells, which either did not express B-Raf or showed decreased Ras activation. Small interfering RNA-mediated depletion of B-Raf in MPNSTs also induced a G(1) cell cycle arrest in these cells, with a marked inhibition of cyclin D1 expression and Rb phosphorylation, whereas depletion of C Raf did not affect either. With growth inhibition at the low nanomolar range, sorafenib, by inhibiting the mitogen-activated protein kinase pathway, may prove to be a novel therapy for patients with MPNST. PMID- 18413801 TI - The proximal promoter region of the human vascular endothelial growth factor gene has a G-quadruplex structure that can be targeted by G-quadruplex-interactive agents. AB - Previous studies on the functional analysis of the human vascular endothelial growth factor (VEGF) promoter using the full-length VEGF promoter reporter revealed that the proximal 36-bp region (-85 to -50 relative to transcription initiation site) is essential for basal or inducible VEGF promoter activity in several human cancer cells. This region consists of a polypurine (guanine) tract that contains four runs of at least three contiguous guanines separated by one or more bases, thus conforming to a general motif capable of forming an intramolecular G-quadruplex. Here, we show that the G-rich strand in this region is able to form an intramolecular propeller-type parallel-stranded G-quadruplex structure in vitro by using the electrophoretic mobility shift assay, dimethyl sulfate footprinting technique, the DNA polymerase stop assay, circular dichroism spectroscopy, and computer-aided molecular modeling. Two well-known G-quadruplex interactive agents, TMPyP4 and Se2SAP, stabilize G-quadruplex structures formed by this sequence in the presence of a potassium ion, although Se2SAP is at least 10-fold more effective in binding to the G-quadruplex than TMPyP4. Between these two agents, Se2SAP better suppresses VEGF transcription in different cancer cell lines, including HEC1A and MDA-MB-231. Collectively, our results provide evidence that specific G-quadruplex structures can be formed in the VEGF promoter region, and that the transcription of this gene can be controlled by ligand-mediated G quadruplex stabilization. Our results also provide further support for the idea that G-quadruplex structures may play structural roles in vivo and therefore might provide insight into novel methodologies for rational drug design. PMID- 18413803 TI - Cyclooxygenase-2 dependent and independent antitumor effects induced by celecoxib in urinary bladder cancer cells. AB - Transitional cell carcinoma of the urinary bladder is the second most common genitourinary malignancy in people in the United States. Cyclooxygenase-2 (COX-2) is overexpressed in bladder cancer. COX-2 inhibitors have had antitumor activity against bladder cancer, but the mechanisms of action are unclear. Clinically relevant concentrations of COX-2 inhibitors fail to inhibit proliferation in standard in vitro assays. In pilot experiments, different culture conditions [standard monolayer, modified monolayer, soft agar, collagen, and poly(2 hydroxyethyl methacrylate)-coated plates] were assessed to determine conditions suitable for the study of COX inhibitor growth-inhibitory effects. This was followed by studies of the effects of clinically relevant concentrations of a selective COX-2 inhibitor (celecoxib) on urinary bladder cancer cell lines (HT1376, TCCSUP, and UMUC3). Celecoxib (3-fold increase in the half-life of NAG-1 mRNA in a p38 kinase-dependent manner and transient transfection experiment showed that VES stabilizes NAG-1 mRNA through AU-rich elements in 3'-untranslated region of NAG-1 mRNA. The inhibition of NAG-1 expression by small interfering RNA significantly blocked VES-induced poly(ADP-ribose) polymerase cleavage, suggesting that NAG-1 may play an important role in VES-induced apoptosis. These results indicate that VES-induced expression of NAG-1 mRNA/protein is regulated by transcriptional/post transcriptional mechanism in a p38 kinase-dependent manner and NAG-1 can be chemopreventive/therapeutic target in prostate cancer. PMID- 18413811 TI - WTp53 induction does not override MTp53 chemoresistance and radioresistance due to gain-of-function in lung cancer cells. AB - New molecular cancer treatment strategies aim to reconstitute wild-type p53 (WTp53) function in mutant p53 (MTp53)-expressing tumors as a means of resensitizing cells to chemotherapy or radiotherapy. The success of this approach may depend on whether MTp53 proteins are acting in a dominant-negative or independent gain-of-function mode. Herein, we describe an isogenic, temperature sensitive p53 model (p53(A138V)) in p53-null human H1299 lung cancer cells in which WTp53 can be selectively coexpressed with a temperature-sensitive MTp53 allele (A138V) during initial DNA damage and subsequent DNA repair. Cells expressing MTp53 alone or coexpressing induced WTp53 and MTp53 were tested for p53 transcription, G(1) and G(2) cell cycle checkpoints, apoptosis, and long-term clonogenic survival following DNA damage. Transient transfection of WTp53 into H1299 cells, or shift-down of H1299-p53(A138V) stable transfectants to 32 degrees C to induce WTp53, led to increased p21(WAF1) expression and G(1) and G(2) arrests following DNA damage but did not increase BAX expression or apoptosis. In contrast, both transient and stable expression of the p53(A138V) mutant in p53 null H1299 cells (e.g. testing gain-of-function) at 37 degrees C blocked p21(WAF1) induction following DNA damage. Cell death was secondary to mitotic catastrophe and/or tumor cell senescence. Overexpression of WTp53 did not resensitize resistant MTp53-expressing cells to ionizing radiation, cisplatinum, or mitomycin C. Our results suggest that human MTp53 proteins can lead to resistant phenotypes independent of WTp53-mediated transcription and checkpoint control. This should be considered when using p53 as a prognostic factor and therapeutic target. PMID- 18413812 TI - Nutlin-3 radiosensitizes hypoxic prostate cancer cells independent of p53. AB - Nutlin-3 is a small-molecule inhibitor that acts to inhibit MDM2 binding to p53 and subsequent p53-dependent DNA damage signaling. Whether Nutlin-3 alters cell toxicity following DNA damage under oxic versus hypoxic conditions has not been studied. The potential radiosensitization (0-10 Gy) properties of Nutlin-3 (dose range, 2-10 micromol/L for up to 24 h) were investigated in vitro using three prostate cancer cell lines, 22RV1 [wild-type p53 (WTp53)], DU145 (mutated p53), and PC-3 (p53-null) under oxic (21% O(2)), hypoxic (0.2% O(2)), and anoxic (0% O(2)) conditions. As a single agent, Nutlin-3 (2-10 micromol/L) stabilized p53 and p21(WAF) levels and was toxic to WTp53-22RV1 cells (IC(50), 4.3 micromol/L) but had minimal toxicity toward p53-deficient cells (IC(50), >10 micromol/L). When combined with radiation under oxic conditions, Nutlin-3 decreased clonogenic survival in all three cell lines: 22RV1 [sensitizing enhancement ratio (SER), 1.24], DU145 (SER, 1.27), and PC-3 (SER, 1.12). Anoxia induced p53 protein expression in 22RV1 cells and this was augmented by Nutlin-3 treatment. Furthermore, Nutlin-3 was more effective as a radiosensitizer under hypoxic conditions particularly in WTp53-expressing cells: 22RV1 (SER, 1.78), DU145 (SER, 1.31), and PC-3 (SER, 1.28). The decrease in clonogenic survival with Nutlin-3 was not correlated to altered levels of radiation-induced apoptosis within the three cell lines. Our results indicate that Nutlin-3 can act as a radiosensitizer via p53-independent mechanisms under low O(2) levels. Nutlin-3 may be a useful adjunct to improve the therapeutic ratio using precision radiotherapy targeted to hypoxic cells and warrants further study in vivo. PMID- 18413813 TI - Targeting Ras in myeloid leukemias. AB - Ras proteins normally relay growth-promoting signals from many activated cell surface receptors, and they are altered by oncogenic point mutations in approximately 30% of human cancers. Activating KRAS and NRAS mutations are especially common in malignancies of the pancreas, lung, and colon, and in myeloid leukemia. Here, we discuss general strategies for targeting hyperactive Ras signaling in cancer cells with specific reference to myeloid malignancies. PMID- 18413814 TI - Epidermal growth factor receptor controls flat dysplastic aberrant crypt foci development and colon cancer progression in the rat azoxymethane model. AB - PURPOSE: Colonic carcinogenesis deranges growth-regulating epidermal growth factor receptors (EGFR). We previously showed that EGFR signals were up-regulated in human aberrant crypt foci (ACF), putative colon cancer precursors. The azoxymethane model of colon cancer recapitulates many aspects of human colonic tumors. Recent studies indicate that flat dysplastic ACF with increased beta catenin are tumor precursors in this model. We asked, therefore, if EGFR signals are required for flat dysplastic ACF development and cancer progression. EXPERIMENTAL DESIGN: Rats received azoxymethane or saline, and standard chow or chow supplemented with gefitinib, an EGFR inhibitor, for 44 weeks. EGFR signals were quantified in normal colon, flat ACF, and tumors by computerized analysis of immunostains and Western blots. K-ras mutations were assessed by PCR and mRNA for egfr ligands by quantitative real-time PCR. RESULTS: EGFR inhibition with gefitinib decreased the incidence of flat dysplastic ACF from 66% to 36% and tumors from 71% to 22% (P < 0.05). This inhibitor also reduced the overexpressions of cyclin D1 and Cox-2 in flat ACF. Furthermore, in flat ACF, EGFR blockade decreased the up-regulation of c-Jun, FosB, phosphorylated active signal transducers and activators of transcription 3, and CCAAT/enhancer binding protein-beta, potential regulators of cyclin D1 and Cox-2. In colonic tumors, EGFR blockade significantly decreased angiogenesis, proliferation, and progression while also increasing apoptosis (P < 0.05). Gefitinib also inhibited the activations of extracellular signal-regulated kinase, Src, and AKT pathways in tumors. CONCLUSIONS: We have shown for the first time that EGFR promotes the development of flat dysplastic ACF and the progression of malignant colonic tumors. Furthermore, we have mechanistically identified several transcription factors and their targets as EGFR effectors in colonic carcinogenesis. PMID- 18413815 TI - Loss of complex I due to mitochondrial DNA mutations in renal oncocytoma. AB - PURPOSE: Many solid tumors exhibit abnormal aerobic metabolism characterized by increased glycolytic capacity and decreased cellular respiration. Recently, mutations in the nuclear encoded mitochondrial enzymes fumarate hydratase and succinate dehydrogenase have been identified in certain tumor types, thus demonstrating a direct link between mitochondrial energy metabolism and tumorigenesis. Although mutations in the mitochondrial genome (mitochondrial DNA, mtDNA) also can affect aerobic metabolism and mtDNA alterations are frequently observed in tumor cells, evidence linking respiratory chain deficiency in a specific tumor type to a specific mtDNA mutation has been lacking. EXPERIMENTAL DESIGN: To identify mitochondrial alterations in oncocytomas, we investigated the activities of respiratory chain enzymes and sequenced mtDNA in 15 renal oncocytoma tissues. RESULTS: Here, we show that loss of respiratory chain complex I (NADH/ubiquinone oxidoreductase) is associated with renal oncocytoma. Enzymatic activity of complex I was undetectable or greatly reduced in the tumor samples (n = 15). Blue Native gel electrophoresis of the multisubunit enzyme complex revealed a lack of assembled complex I. Mutation analysis of the mtDNA showed frame-shift mutations in the genes of either subunit ND1, ND4, or ND5 of complex I in 9 of the 15 tumors. CONCLUSION: Our data indicate that isolated loss of complex I is a specific feature of renal oncocytoma and that this deficiency is frequently caused by somatic mtDNA mutations. PMID- 18413816 TI - Disrupted expression of CXCL5 in colorectal cancer is associated with rapid tumor formation in rats and poor prognosis in patients. AB - PURPOSE: We isolated a subline (CC531M) from the CC531S rat colon carcinoma cell line, which grows and metastasizes much more rapidly than CC531S. We found, using RNA expression profiling, that one of the major changes in the CC531M cell line was a 5.8-fold reduction of the chemokine CXCL5. The purpose of this study was to determine the effect of CXCL5 expression on colorectal tumor growth and metastasis. EXPERIMENTAL DESIGN: CC531 clones were generated with either knockdown or restored expression of CXCL5. These clones were inoculated in the liver of rats. In addition, in two independent cohorts of colorectal cancer patients, the level of CXCL5 expression was determined and associated to clinical variables. RESULTS: Knockdown of CXCL5 expression in CC531S resulted in rapid tumor growth and increased number of metastasis, whereas restored expression of CXCL5 in CC531M resulted in a return of the "mild" tumor growth pattern of the parental cell line CC531S. In vitro, no difference was found in proliferation rate between clones with either high or low expression of CXCL5, suggesting that environmental interactions directed by CXCL5 determine tumor outgrowth. Finally, the importance of our findings was established for patients with colorectal cancer. We found that low expression of CXCL5 was significantly associated with poor prognosis for colorectal cancer patients. CXCL5 showed a trend (P = 0.05) for a positive correlation with intratumoral CD8(+) T-cell infiltration, suggesting a possible explanation for the observed poorer prognosis. CONCLUSIONS: Our results show that CXCL5 is important in growth and development of colorectal cancer, implicating a future role in both cancer therapy and diagnosis. PMID- 18413817 TI - KIT mutations induce intracellular retention and activation of an immature form of the KIT protein in gastrointestinal stromal tumors. AB - PURPOSE: Gastrointestinal stromal tumors (GIST) are frequently associated with gain-of-function mutations of KIT, which can be inhibited by imatinib both in vitro and in vivo. The survival of patients with GIST, following imatinib therapy, has been correlated with the nature of mutations but not with KIT expression. EXPERIMENTAL DESIGN: Subcellular localization, activation, and trafficking of the mature and the immature forms of KIT were investigated in GIST samples and in NIH3T3 cells infected with two different GIST-type exon 11-mutated human KIT cDNA. RESULTS: Paranuclear dot expression of KIT was more frequent in GISTs with homozygous KIT mutations than in those with heterozygous (P = 0.01) or no mutations (P < 0.01). Activation of the immature 125 kDa form of KIT was detected in most GISTs with KIT mutations but not in GISTs without KIT mutations. In NIH3T3 cells, mutant KIT was mainly retained within endoplasmic reticulum and Golgi compartments in an immature constitutively phosphorylated form, whereas the wild-type KIT was expressed at the plasma membrane, in a mature nonphosphorylated form. Imatinib-induced inhibition of the phosphorylation of immature and mature mutant KIT proteins resulted in the restoration of KIT expression at the cell surface. CONCLUSIONS: These results show that GIST-type KIT mutations induce an activation-dependent alteration of normal maturation and trafficking, resulting in the intracellular retention of the activated kinase within the cell. These observations likely account for the absence of correlation between response to imatinib and KIT expression using immunohistochemistry and may deserve to be investigated in other tyrosine kinase-activated tumors. PMID- 18413818 TI - Ornithine decarboxylase G316A genotype is prognostic for colorectal adenoma recurrence and predicts efficacy of aspirin chemoprevention. AB - PURPOSE: The chemopreventive activity of aspirin in colorectal neoplasia may be explained in part by its effect on polyamine metabolism. The ornithine decarboxylase (ODC) G316A polymorphism affects polyamine metabolism through altered expression of ODC. We investigated the influence of ODC G316A on the chemopreventive activity of aspirin in colorectal adenoma (CRA) recurrence. EXPERIMENTAL DESIGN: We genotyped ODC G316A in 546 individuals in the United Kingdom Colorectal Adenoma Prevention trial of aspirin for CRA recurrence prevention and pooled our findings with data from two other randomized intervention trials. RESULTS: The United Kingdom Colorectal Adenoma Prevention participants with homozygous ODC 316AA genotype were at reduced CRA recurrence risk [relative risk (RR), 0.43; 95% confidence interval (95% CI), 0.16-1.15], particularly if also exposed to aspirin (RR, 0.24; 95% CI, 0.03-1.71). In the pooled analysis of 2,207 individuals, those with homozygous ODC 316AA genotype were at significantly reduced CRA recurrence risk (RR, 0.68; 95% CI, 0.47-0.99). Following stratification by genotype and aspirin exposure, individuals with homozygous wild-type or heterozygous genotypes derived modest benefit from aspirin (RR, 0.85; 95% CI, 0.72-1.01), whereas in those with both ODC 316AA genotype and aspirin exposure recurrence risk was halved (RR, 0.52; 95% CI, 0.29 0.91). CONCLUSION: The ODC G316A genotype is prognostic for CRA recurrence and predictive of an enhanced response to aspirin in preventing recurrence. This variant has the potential to be a clinically useful genetic marker to identify individuals likely to derive the greatest benefit from aspirin chemoprevention. PMID- 18413819 TI - Inverse correlation between tumoral indoleamine 2,3-dioxygenase expression and tumor-infiltrating lymphocytes in endometrial cancer: its association with disease progression and survival. AB - PURPOSE: Tumor escape from host immune systems is a crucial mechanism for disease progression. We recently showed that the immunosuppressive enzyme indoleamine 2,3 dioxygenase (IDO) is a prognostic indicator for endometrial cancer. The purpose of the present study was to investigate the relationship between IDO expression and tumor-infiltrating lymphocytes (TIL) or natural killer (NK) cells and to clarify their prognostic effect in endometrial cancer. EXPERIMENTAL DESIGN: Immunohistochemical staining for IDO expression in endometrial cancer tissues (n = 65) was done. Tumor-infiltrating CD3+ and CD8+ lymphocytes, as well as CD57+ NK cells, were counted in serial tissue sections. RESULTS: High IDO expression in tumor cells was found in 32 of 65 cases and was positively correlated with myometrial invasion, nodal metastasis, and lymph-vascular space involvement. We also found a significant correlation between high IDO expression and reduced numbers of CD3+, CD8+, and CD57+ cells infiltrating into both the tumor epithelium and stroma. Patients with high IDO expression, a low number of stromal CD3 (<60), low intraepithelial CD8 (<25), or low stromal CD8 (<40) had significantly impaired progression-free survival. On multivariate analysis, IDO expression and the number of stromal CD3+ TILs were independent prognostic factors for impaired progression-free survival. CONCLUSIONS: Tumoral IDO expression correlated with a reduced number of TILs and NK cells in endometrial cancer, possibly contributing to disease progression and impaired clinical outcome. These findings suggest that targeting IDO to restore host antitumor immunity may be a therapeutic strategy for endometrial cancer. PMID- 18413820 TI - Increased expression of Id family proteins in small cell lung cancer and its prognostic significance. AB - PURPOSE: To study the molecular pathology of human small cell lung cancer (SCLC), molecular biology approaches were used to identify genes involved in malignant progression of the cancer cells. EXPERIMENTAL DESIGN: Microquantity differential display was used initially to identify genes expressed differentially between normal and malignant cell lines. The differences were verified by Western blot. Immunohistochemical analysis was done on paired normal and malignant lung tissues and on tissues taken by biopsy to assess the expression status of candidate genes and their prognostic significance. RESULTS: Inhibitor of DNA/differentiation (Id)1 gene was up-regulated in SCLC cells. Levels of Id1 in 8 of 10 cell lines were increased by 1.7- to 21.4-fold when compared with the benign cells. A similar increase was also found in levels of Id2 and Id3. On 26 pairs of lung tissues, all four Id proteins were significantly (Wilcoxon Signed Rank Test, P < 0.001-0.005) overexpressed in cytoplasm of the malignant cells. In nuclei of SCLC cells, Id1 expression was significantly reduced, whereas the levels of Id2, Id3, and Id4 were significantly (Wilcoxon Signed Rank Test, P < 0.001) increased. Immunohistochemical staining on biopsy specimens showed that the increased expression of Id2 in cytoplasm of cancer cells, not the other three proteins, was significantly associated with the increased survival of SCLC patients. CONCLUSION: Changed expression profiles of Id proteins may play important roles in malignant progression of SCLC, and the increased Id2 in cytoplasm is a novel prognostic factor to predict the patient outcomes. PMID- 18413821 TI - Up-regulation of peroxiredoxin 1 in lung cancer and its implication as a prognostic and therapeutic target. AB - PURPOSE: Peroxiredoxin 1 and 2 are highly homologous members of the Prx (or Prdx) protein family. Prx1 and Prx2 are elevated in several human cancers, and this seems to confer increased treatment resistance and aggressive phenotypes. This study was undertaken to examine the expression profiles of Prx1 and Prx2 in non small cell lung cancer (NSCLC), and to test their prognostic value in predicting patient survival. EXPERIMENTAL DESIGN: To gain insight into the regulatory mechanisms of Prx1 and Prx2 expression in NSCLC, their respective transcript profiles were examined in NSCLC cell lines from the NCI-60 panel Affymetrix database sets, and the promoter compositions of the two genes were investigated using computer-based multiple sequence alignment analyses. Immunohistochemical analyses of Prx1 and Prx2 were done on a total of 235 NSCLC specimens with stage I through IV disease. The expression profiles of Prx1 and Prx2 in tumor specimens, and their associations with survival, were investigated. RESULTS AND CONCLUSION: The levels of prx1 transcript were higher than those of prx2 in NSCLC cell lines, and the upstream regulatory sequences of the two genes display striking differences. The relative risk of death increased as Prx1 expression levels increased (P = 0.036) in a multivariate Cox model, independent of other clinicopathologic variables associated with survival. No statistically significant correlation was observed between Prx2 and survival. These results suggest that Prx1 may possess unique functions and regulatory mechanisms in NSCLC which are not shared with Prx2, and that Prx1 may serve as a new prognostic biomarker and therapeutic target in NSCLC. PMID- 18413822 TI - Clinical significance of high mobility group A2 in human gastric cancer and its relationship to let-7 microRNA family. AB - PURPOSE: The high mobility group A2 (HMGA2) nonhistone chromosomal protein can modulate transcription by altering chromatin architecture. HMGA2 is highly expressed during embryogenesis and in various benign and malignant tumors. Recent studies report that HMGA2 is negatively regulated by the let-7 microRNA (miRNA) family. However, no studies have examined the clinical significance of HMGA2 and its relationship to the let-7 miRNA family in gastric cancer. EXPERIMENTAL DESIGN: Using quantitative real-time reverse transcription-PCR, we analyzed HMGA2 expression with respect to various clinicopathologic factors in 110 patients with gastric cancer. We also did an association study comparing HMGA2 expression and let-7 miRNA family expression in gastric cancer. RESULTS: Expression of HMGA2 in cancerous tissues was significantly higher than in noncancerous tissues (P < 0.05). Elevated HMGA2 expression was significantly correlated with serosal invasion (P < 0.05) and poor clinical prognosis (P < 0.05). A multivariate analysis showed that HMGA2 expression status was an independent prognostic factor (P < 0.05). An inverse correlation between HMGA2 and let-7a was found in gastric cancer cell lines (P = 0.08). The expressions of let-7a, let-7b, and let-7c in gastric cancer patients with low HMGA2 expression were significantly higher than those with high HMGA2 expression (P < 0.05). CONCLUSIONS: High expression of HMGA2 in gastric cancer correlates with tumor invasiveness and is an independent prognostic factor. Furthermore, our findings suggest that HMGA2 is negatively regulated by the let-7 miRNA family in human gastric cancer. PMID- 18413823 TI - Vascularization in primary breast carcinomas: its prognostic significance and relationship with tumor cell dissemination. AB - PURPOSE: The interaction between tumor cells, stroma, and endothelial cells is important for the dissemination of tumor cells. The aim of the present study is to examine vascularity in primary breast carcinomas and its prognostic significance and relationship with tumor cell dissemination. EXPERIMENTAL DESIGN: A total of 498 invasive breast carcinomas were analyzed. Representative tumor sections were stained for CD34 and CD105, and vascularity was quantified by the Chalkley method. The relationship between Chalkley counts, vascular invasion, disseminated tumor cells (DTC) in the bone marrow, other clinicopathologic variables, and clinical outcome was evaluated. RESULTS: High vascular grades determined by Chalkley counts were significantly associated with shorter distant disease-free survival and breast cancer-specific survival in all patients (P < 0.001, log-rank) and in node-negative patients not receiving adjuvant systemic therapy (P < 0.05). In multivariate analysis, both CD34 and CD105 Chalkley counts showed prognostic significance for distant disease-free survival (P = 0.014 and P = 0.026), whereas CD34 also showed prognostic significance for breast cancer specific survival (P = 0.007). Vascular invasion and DTCs in the bone marrow showed independent prognostic significance. DTC did not discriminate survival for CD34 low Chalkley counts, whereas a very poor prognosis was observed for DTC positive patients with high CD34 counts. In node-negative patients not receiving systemic chemotherapy, high CD34 and high CD105 counts in combination identified patients with unfavorable outcome, as opposed to all other CD34/CD105 combinations. CONCLUSIONS: Improved identification of risk groups could be obtained by adding CD34 and CD105 vascular analysis to DTC, vascular invasion, and other primary tumor factors. This may facilitate the selection of candidates for adjuvant systemic therapy. PMID- 18413824 TI - Amphiregulin is a promising prognostic marker for liver metastases of colorectal cancer. AB - PURPOSE: Aberrant activation of epidermal growth factor receptors (EGFR/HER1) by ligand stimulation or heterodimerization with human epidermal growth factor 2 (HER2) is considered to play an important role in the development of colorectal carcinoma. Amphiregulin (AR) is a ligand of EGFR that might be related to the development and progression of gastrointestinal tumors. The aim of this study was to determine the AR, EGFR, and HER2 protein expression levels and to evaluate their prognostic relevance to the clinical course of colorectal cancer. EXPERIMENTAL DESIGN: The AR, EGFR, and HER2 protein levels in primary tumors of colorectal cancer (n = 106) were examined using immunohistochemistry. Metastatic sites in liver specimens (n = 16) were also analyzed in the same manner. RESULTS: Thirteen (81.6%) metastatic lesions of the liver stained positive for AR. Among the primary lesions of colorectal cancer, 58 (54.7%) stained positive for AR, 13 (12.3%) stained positive for EGFR, and 5 (4.7%) stained positive for HER2. When the relationships between each protein expression level and the clinicopathologic factors were examined, only the AR expression level was significantly related to liver metastasis (P = 0.0296). A multivariate analysis of liver metastasis proved that AR expression was an independent prognostic factor of liver metastasis from colorectal cancer (P = 0.0217). CONCLUSIONS: AR expression in primary lesions of colorectal cancer is an important predictive marker of liver metastasis. PMID- 18413825 TI - The use of magnetic resonance imaging to noninvasively detect genetic signatures in oligodendroglioma. AB - BACKGROUND: Some patients with low-grade glioma have extraordinarily long survival times; current, early treatment does not prolong their lives. For this reason, therapies that sometimes have neurologic side effects are often deferred intentionally. METHODS: In a study of oligodendrogliomas, we used a quantitative method of MR analysis based on the S-transform to investigate whether codeletion of chromosomes 1p and 19q, a marker of good prognosis, could be predicted accurately by measuring image texture. RESULTS: Differences in texture were seen between tumors with codeletion of chromosomes 1p and 19q and those with intact 1p and 19q alleles on contrast-enhanced T1-weighted and T2-weighted MR images. Quantitative MR texture on T2 images predicted codeletion of chromosomes 1p and 19q with high sensitivity and specificity. CONCLUSIONS: This new method of MR image interpretation may have the potential to augment the diagnostic assessment of patients with suspected low-grade glioma. PMID- 18413826 TI - Activation of placenta-specific transcription factor distal-less homeobox 5 predicts clinical outcome in primary lung cancer patients. AB - PURPOSE AND EXPERIMENTAL DESIGN: To identify novel biomarkers and therapeutic targets for lung cancers, we screened for genes that were highly transactivated in lung cancers using a cDNA microarray representing 27,648 genes. DLX5 gene, a member of the human distal-less homeobox transcriptional factor family that is expressed during early embryonic development, was found to be overexpressed in the great majority of lung cancers. Tissue microarray consisting of archival non small cell lung cancer samples from 369 patients was applied to examine the clinicopathologic significance of DLX5 protein. A role of DLX5 in cancer cell growth and/or survival was investigated through small interfering RNA experiments. RESULTS: Northern blot and immunohistochemical analyses detected expression of DLX5 only in placenta among 23 normal tissues examined. Immunohistochemical analysis showed that positive immunostaining of DLX5 was correlated with tumor size (pT classification; P = 0.0053) and poorer prognosis of non-small cell lung cancer patients (P = 0.0045). It was also shown to be an independent prognostic factor (P = 0.0415). Treatment of lung cancer cells with small interfering RNAs for DLX5 effectively knocked down its expression and suppressed cell growth. CONCLUSIONS: These data implied that DLX5 is useful as a target for the development of anticancer drugs and cancer vaccines as well as for a prognostic biomarker in clinic. PMID- 18413827 TI - A genotype-phenotype examination of cyclin D1 on risk and outcome of squamous cell carcinoma of the head and neck. AB - PURPOSE: The variant allele of CCND1 G870A encodes a splice variant of the cyclin D1 protein, which possesses an increased half-life. To confirm the phenotypic effect of the variant allele, we examined the immunohistochemical staining pattern of the protein in tumors from a case population of head and neck squamous cell carcinoma (HNSCC) and compared it with the genotype of these individuals. We also examined how this genotype was associated with the risk of HNSCC and if this genotype-phenotype association was related to patient outcome. EXPERIMENTAL DESIGN: In a population-based case-control study of 698 cases and 777 controls, we both genotyped all participants for the CCND1 gene and did immunohistochemical staining of the cyclin D1 protein in the HNSCC tumors. RESULTS: The variant AA genotype was significantly associated with positive immunohistochemical staining (P < 0.02), and this variant genotype was associated with a significantly elevated odds ratio of 1.5 (95% confidence interval, 1.1-2.0) for HNSCC overall, with risk greatest in oral and laryngeal sites. Positive immunohistochemical staining was inversely related to human papillomavirus 16 DNA present in the tumor (P < 0.03). The AA genotype and superpositive immunohistochemical staining for cyclin D1 also had independent and significant effects on patient survival. CONCLUSIONS: These results strongly suggest that this splice variant, when present in two copies, is a significant predictor of both the occurrence of HNSCC as well as patient survival after treatment. These data further indicate that this variant protein is an important determinant of individual response to therapy for this disease. PMID- 18413828 TI - Urinary biomarkers predict brain tumor presence and response to therapy. AB - PURPOSE: A major difficulty in treating brain tumors is the lack of effective methods of identifying novel or recurrent disease. In this study, we have evaluated the efficacy of urinary matrix metalloproteinases (MMP) as diagnostic biomarkers for brain tumors. EXPERIMENTAL DESIGN: Urine, cerebrospinal fluid, and tissue specimens were collected from patients with brain tumors. Zymography, ELISA, and immunohistochemistry were used to characterize the presence of MMP-2, MMP-9, MMP-9/neutrophil gelatinase-associated lipocalin (NGAL), and vascular endothelial growth factor (VEGF). Results were compared between age- and sex matched controls and subjected to univariate and multivariate statistical analyses. RESULTS: Evaluation of a specific panel of urinary biomarkers by ELISA showed significant elevations of MMP-2, MMP-9, MMP-9/NGAL, and VEGF (all P < 0.001) in samples from brain tumor patients compared with controls. Multiplexing MMP-2 and VEGF provided superior accuracy compared with any other combination or individual biomarker. Receiver-operating characteristics curves for MMP-2 and VEGF showed excellent discrimination. Immunohistochemistry identified these same proteins in the source tumor tissue. A subset of patients with longitudinal follow-up revealed subsequent clearing of biomarkers after tumor resection. CONCLUSION: We report, for the first time, the identification of a panel of urinary biomarkers that predicts the presence of brain tumors. These biomarkers correlate with presence of disease, decrease with treatment, and can be tracked from source tissue to urine. These data support the hypothesis that urinary MMPs and associated proteins are useful predictors of the presence of brain tumors and may provide a basis for a novel, noninvasive method to identify new brain tumors and monitor known tumors after treatment. PMID- 18413829 TI - Clinical, radiographic, and biochemical characterization of multiple myeloma patients with osteonecrosis of the jaw. AB - PURPOSE: Osteonecrosis of the jaw (ONJ) has been reported in patients with a history of aminobisphosphonate use. This study was conducted in order to define ONJ clinically and radiographically and gain insights into its pathophysiology. EXPERIMENTAL DESIGN: Eleven multiple myeloma (MM) patients with ONJ were included in the study. Patients underwent clinical, biochemical, radiographic, and molecular profiling. Ten MM patients on aminobisphosphonates without ONJ and five healthy volunteers were used as controls for biochemical and molecular studies. RESULTS: MM patients with ONJ were treated with either pamidronate (n = 3), zoledronate (n = 4), or both agents sequentially (n = 4) for a mean of 38.7 months. Radiographic studies showed bone sclerosis and fragmentation on plain films and computerized tomography. Quantitative regional analysis of NaF-PET and FDG-PET scans confirmed an increased standardized uptake value (SUVmax) in areas of ONJ. The target to background ratio of SUVmax was significantly greater for NaF-PET compared with FDG-PET scan. Biochemical bone marker data and transcriptional profiling studies showed that genes and proteins involved in osteoblast and osteoclast signaling cascades were significantly down-regulated in patients with ONJ. CONCLUSIONS: ONJ was associated with a mean duration of 38.7 months of aminobisphosphonate exposure. Radiographic and functional imaging confirmed sites of clinically established ONJ. Gene and protein studies are consistent with altered bone remodeling, evidenced by suppression of both bone resorption and formation. PMID- 18413831 TI - Prediction of broad spectrum resistance of tumors towards anticancer drugs. AB - PURPOSE: Drug resistance is a major obstacle in cancer chemotherapy. Although the statistical probability of therapeutic success is known for larger patient groups from clinical therapy trials, it is difficult to predict the individual response of tumors. The concept of individualized therapy aims to determine in vitro the drug response of tumors beforehand to choose effective treatment options for each individual patient. EXPERIMENTAL DESIGN: We analyzed the cross-resistance profiles of different tumor types (cancers of lung, breast, and colon, and leukemia) towards drugs from different classes (anthracyclines, antibiotics, Vinca alkaloids, epipodophyllotoxins, antimetabolites, and alkylating agents) by nucleotide incorporation and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays. Hierarchical cluster analysis and COMPARE analyses were applied. RESULTS: Tumors exert broad resistance profiles, e.g., tumors resistant to one drug tend to also be resistant to other drugs, whereas sensitive tumors reveal sensitivity towards many drugs. Interestingly, the broad spectrum resistance phenotype could reliably be predicted by doxorubicin alone. Expression of the ATP-binding cassette transporter P-glycoprotein (ABCB1, MDR1) and the proliferative activity of tumors were identified as underlying mechanisms of broad spectrum resistance. To find novel compounds with activity against drug resistant tumors, a database with 2,420 natural products was screened for compounds acting independent of P-glycoprotein and the proliferative state of tumor cells. CONCLUSIONS: Tumors exert cross-resistance profiles much broader than the classical multidrug resistance phenotype. Broad spectrum resistance can be predicted by doxorubicin due to the multifactorial mode of action of this drug. Novel cytotoxic compounds from natural resources might be valuable tools for strategies to bypass broad spectrum resistance. PMID- 18413830 TI - Understanding the biological basis of autofluorescence imaging for oral cancer detection: high-resolution fluorescence microscopy in viable tissue. AB - PURPOSE: Autofluorescence imaging is increasingly used to noninvasively identify neoplastic oral cavity lesions. Improving the diagnostic accuracy of these techniques requires a better understanding of the biological basis for optical changes associated with neoplastic transformation in oral tissue. EXPERIMENTAL DESIGN: A total of 49 oral biopsies were considered in this study. The autofluorescence patterns of viable normal, benign, and neoplastic oral tissue were imaged using high-resolution confocal fluorescence microscopy. RESULTS: The autofluorescence properties of oral tissue vary significantly based on anatomic site and pathologic diagnosis. In normal oral tissue, most of the epithelial autofluorescence originates from the cytoplasm of cells in the basal and intermediate regions, whereas structural fibers are responsible for most of the stromal fluorescence. A strongly fluorescent superficial layer was observed in tissues from the palate and the gingiva, which contrasts with the weakly fluorescent superficial layer found in other oral sites. Upon UV excitation, benign inflammation shows decreased epithelial fluorescence, whereas dysplasia displays increased epithelial fluorescence compared with normal oral tissue. Stromal fluorescence in both benign inflammation and dysplasia drops significantly at UV and 488 nm excitation. CONCLUSION: Imaging oral lesions with optical devices/probes that sample mostly stromal fluorescence may result in a similar loss of fluorescence intensity and may fail to distinguish benign from precancerous lesions. Improved diagnostic accuracy may be achieved by designing optical probes/devices that distinguish epithelial fluorescence from stromal fluorescence and by using excitation wavelengths in the UV range. PMID- 18413832 TI - Pathologic complete response to neoadjuvant chemotherapy of breast carcinoma is associated with the disappearance of tumor-infiltrating foxp3+ regulatory T cells. AB - PURPOSE: T-cell infiltration is associated with good tumor prognosis in many cancers. To assess the capacity of neoadjuvant chemotherapy to affect T-cell infiltration in breast cancer, we evaluated CD3 and CD8 infiltrates, and the Foxp3 immunosuppressive T cells. EXPERIMENTAL DESIGN: CD3+, CD8+, and Foxp3+ cell infiltrates were detected by immunohistochemistry in a series of 56 breast cancer patients before and after the end of neoadjuvant chemotherapy. RESULTS: Poor prognostic factors (negative hormonal receptors, high tumor grade, and nodal involvement) were associated with a significantly higher number of CD3, CD8, and Foxp3 infiltrates before the beginning of chemotherapy. Chemotherapy resulted in a decrease in Foxp3 infiltrates, whereas the level of CD8 and CD3 infiltrates remained unchanged. Pathologic complete responses (pCR) had a drastic decrease of Foxp3+ cells, whereas these cells remained elevated in nonresponders. A cutoff criterion that combined high CD8 infiltration and no Foxp3 cell infiltration on surgical specimens is associated with pCR with a sensitivity of 75% and a specificity of 93%. The infiltrate of cytotoxic TiA1 and granzyme B-positive cells was dramatically enhanced after chemotherapy only in patients with pCR. By multivariate analysis, association of a high CD8 infiltration and no Foxp3 infiltration on final histologic specimens were independently associated with pCR. CONCLUSION: These findings indicate that pCR to neoadjuvant chemotherapy is associated with an immunologic profile combining the absence of immunosuppressive Foxp3 cells and the presence of a high number of CD8 T cells and cytotoxic cells. These results argue for the induction of an antitumor immune response by chemotherapy. PMID- 18413833 TI - Effects of a topically applied bioadhesive berry gel on loss of heterozygosity indices in premalignant oral lesions. AB - PURPOSE: The aim of this study was to assess the effects of topical application of a 10% (w/w) freeze-dried black raspberry (FBR) gel on oral intraepithelial neoplasia (IEN) variables that included histologic diagnoses and loss of heterozygosity (LOH) indices. Microsatellite instability and/or LOH at tumor suppressor gene-associated chromosomal loci have been associated with a higher risk for oral IEN progression to oral squamous cell carcinoma. Previously, our laboratories have shown that FBRs are well tolerated and possess potent antioxidant, apoptotic, and differentiation-inducing properties. EXPERIMENTAL DESIGN: Each participant with IEN served as their own internal control. Before treatment, all lesions were photographed, and lesional tissue was hemisected to obtain a pretreatment diagnosis and baseline biochemical and molecular variables. Gel dosing (0.5 g applied four times daily for 6 weeks) was initiated 1 week after the initial biopsy. Genomic DNA was isolated from laser-captured basilar and suprabasilar surface epithelial cells followed by PCR amplification using primer sets that targeted known and presumed tumor suppressor gene loci associated with INK4a/ARF, p53, and FHIT. Allelic imbalance was determined by sequence analysis using normal participant tissues to establish microsatellite marker peak patterns and allele sizes. RESULTS: Confirming earlier phase I data, none of the 27 participants developed FBR gel-associated toxicities. Furthermore, our results show histologic regression in a subset of patients as well as statistically significant reduction in LOH at tumor suppressor gene-associated loci. CONCLUSIONS: These preliminary data suggest that further evaluation of berry gels for oral IEN chemoprevention is warranted. PMID- 18413834 TI - Sunitinib for treatment of advanced renal cell cancer: primary tumor response. AB - PURPOSE: Nephrectomy before immunotherapy in patients with metastatic renal cell cancer (RCC) will improve patient outcome. In addition, the primary tumor is known to be refractory to cytokines. Sunitinib is now approved for treatment of advanced RCC, but its effect on the primary tumor has yet to be reported. EXPERIMENTAL DESIGN: All patients treated with sunitinib for advanced RCC without prior nephrectomy were reviewed and sequential computed tomography scans were evaluated for response in the primary tumor as well as metastases according to Response Evaluation Criteria in Solid Tumors. Volumes of primary tumors and central necrotic areas were measured with the perimeter method. RESULTS: Computed tomography scans were available for evaluation of response in 17 of 22 patients with a primary tumor in situ (1 patient with two primaries). According to Response Evaluation Criteria in Solid Tumors, 4 patients had a partial response, 12 had stable disease, and 1 had progressive disease. The one-dimensional longest diameter of the primary tumor correlated with the volumetric measurements both at baseline and at the time of evaluation of response. Excluding the patient with progressive disease, the median volume reduction was 31% associated with a median increase in the volume of necrosis of 39%. Three patients underwent nephrectomy and tumors showed extensive necrotic areas next to small fields of vital tumor cells. CONCLUSIONS: Sunitinib can induce a significant reduction in volume of primary renal cell tumors. Further trials need to address the role of nephrectomy in advanced RCC patients on sunitinib treatment. PMID- 18413835 TI - Randomized, double-blinded phase II evaluation of docetaxel with or without doxercalciferol in patients with metastatic, androgen-independent prostate cancer. AB - PURPOSE: Docetaxel is standard of care for androgen-independent prostate cancer (AIPC). Doxercalciferol (1 alpha-hydroxyvitamin D2) had modest activity in phase I/II trials. Preclinical data support combining vitamin D analogues with docetaxel to treat AIPC. EXPERIMENTAL DESIGN: Chemotherapy-naive men with metastatic AIPC were randomized 1:1 to receive, on a 4-week cycle, docetaxel (35 mg/m2 i.v., days 1, 8, and 15) with or without doxercalciferol (10 microg orally, days 1-28). The primary end point was prostate-specific antigen (PSA) response. Secondary end points were progression-free survival, overall survival, objective response, and toxicity. Survival was analyzed as intent to treat. RESULTS: Seventy patients were randomized. Median follow-up was 17.6 months (range, 3.3 45.2). PSA response rate was 46.7% [95% confidence interval (95% CI), 30-64] in the doxercalciferol arm and 39.4% (95% CI, 25-56) with placebo (P = 0.560). Median progression-free survival in the doxercalciferol arm was 6.17 months (95% CI, 4.20-10.7) versus 6.20 months (95% CI, 4.83-9.07) with placebo (P = 0.764). Median overall survival in the doxercalciferol arm was 17.8 months (95% CI, 14.9 23.6) versus 16.4 months (95% CI, 11.9-23.8) with placebo (P = 0.383). Twenty four patients in the doxercalciferol arm and 23 in the placebo arm were evaluable for objective response. No complete responses were observed. Partial objective response rate was 12.5% with doxercalciferol versus 8.7% with placebo (P = 0.672). Rate of grade > or =3 toxicity was 46% with doxercalciferol versus 42% with placebo (P = 0.785). CONCLUSIONS: Daily doxercalciferol with weekly docetaxel did not enhance PSA response rate or survival. Toxicity was similar between arms. Despite the disappointing results of this study, other vitamin D analogues remain under active investigation. PMID- 18413836 TI - A phase I biological study of MG98, an oligodeoxynucleotide antisense to DNA methyltransferase 1, in patients with high-risk myelodysplasia and acute myeloid leukemia. AB - PURPOSE: Epigenetic silencing via aberrant promoter DNA hypermethylation of normal genes has been described as a leukemogenic mechanism in myelodysplastic syndromes (MDS) and acute myeloid leukemias (AML). We hypothesized that MG98, an oligonucleotide antisense to DNA methyltransferase 1 (DNMT1), could reverse malignant phenotypes by down-regulating DNMT1 and inducing reexpression of hypermethylated genes. This phase I study was conducted to determine a biologically effective dose and describe the safety of MG98 in MDS/AML. EXPERIMENTAL DESIGN: Twenty-three patients with MDS (n = 11) and AML (n = 12) were enrolled. Biologically effective dose was defined as the dose at which > or =50% of patients experienced >50% reduction in DNMT1 expression with acceptable toxicity. Escalating doses of MG98 were administered according to two schedules (2-hour i.v. bolus followed by 5-day continuous i.v. infusion every 14 days, or 14-day continuous i.v. infusion every 21 days). RESULTS: DNMT1 down-regulation was observed in 8 patients. However, biologically effective dose was not reached. Reexpression of target genes (P15, WIT1, and ER) was observed in 12 patients but did not correlate with DNMT1 down-regulation. Escalation was stopped due to dose limiting toxicities (bone pain, nausea, and fever). No objective clinical response was observed. Disease stabilization occurred in 6 (26%) patients. CONCLUSIONS: No pharmacodynamic or clinical activity was observed at MG98 doses and schedules administered. Despite this, pursuing DNMT1 down-regulation remains a sound approach for targeting aberrant epigenetics in AML/MDS. Future studies with different formulation and/or doses and schedules will be required to ensure efficient MG98 intracellular uptake and fully evaluate its therapeutic potential. PMID- 18413837 TI - Treatment of cancer with a novel dual-targeted conditionally replicative adenovirus armed with mda-7/IL-24 gene. AB - PURPOSE: Recent studies showed that oncolytic adenoviruses not only have capacity for destruction of tumors but also can be used as potential vectors to express therapeutic genes for therapy of cancer. However, better specificity and mode of transgene expression are required to improve the efficacy and safety if this vector is applied for clinical application. EXPERIMENTAL DESIGN: In this study, we have created adenoviral replication-based transgene expression system by replacement of 6.7K/gp19K of E3 genes with EGFP and IL-24 genes so that expression of transgenes should be controlled by adenoviral E3 promoter. Transgene expression, viral replication capacity, and cytotoxicity have been studied in tumor and normal cells. Antitumor efficacy was evaluated in animal model with established tumor. RESULTS: Our data showed that expression of IL-24 could be detected at 6 h and reached the maximal level at 48 h after infection in tumor cells. The expression level was 14 times higher than that induced by cytomegalovirus promoter. Low level of IL-24 could be detected in normal cells only until 72 h after infection. The substitution of 6.7K/gp19K of E3 genes with transgenes did not affect viral replication in tumor cells. Strong cytotoxicity was observed only in tumor cells after infection with AdCN205-IL-24. Treatment of the established tumors induced high level of local expression of IL-24 in tumor cells and resulted in tumor regression. CONCLUSIONS: Our data showed that AdCN205 IL-24 can provide potent and safe vector for the therapy of cancer. PMID- 18413838 TI - Inhibiting fatty acid synthase for chemoprevention of chemically induced lung tumors. AB - PURPOSE: Fatty acid synthase (FAS) is overexpressed in lung cancer, and we have investigated the potential use of FAS inhibitors for chemoprevention of lung cancer. EXPERIMENTAL DESIGN: Expression of FAS was evaluated in preinvasive human lung lesions (bronchial squamous dysplasia and atypical adenomatous hyperplasia) and in murine models of lung tumorigenesis [4-(methylnitrosamino)-I-(3-pyridyl)-1 butanone-induced and urethane-induced lung tumors in A/J mice]. Then, the ability of pharmacologic inhibitors of FAS to prevent development of the murine tumors was investigated. Finally, the effect of the FAS inhibitor treatment of levels of phosphorylated Akt in the murine tumors was evaluated by immunohistochemistry. RESULTS: Immunohistochemical studies show that human bronchial dysplasia and atypical adenomatous hyperplasia express high levels of FAS compared with normal lung tissues, suggesting that FAS might be a target for intervention in lung carcinogenesis. FAS is also expressed at high levels in chemically induced murine lung tumors, and the numbers and sizes of those murine tumors are significantly reduced by treating carcinogen-exposed mice with pharmacologic inhibitors of FAS, C75 and C93. C93 treatment is associated with reduced levels of phosphorylated Akt in tumor tissues, suggesting that inhibition of this signal transduction pathway might be involved in the chemopreventative activity of this compound. CONCLUSIONS: We conclude that increased levels of FAS are common in human preinvasive neoplasia of the lung. Based on studies in mouse models, it seems that inhibiting FAS is an effective strategy in preventing and retarding growth of lung tumors that have high expression of this enzyme. PMID- 18413839 TI - EXEL-7647 inhibits mutant forms of ErbB2 associated with lapatinib resistance and neoplastic transformation. AB - PURPOSE: Mutations associated with resistance to kinase inhibition are an important mechanism of intrinsic or acquired loss of clinical efficacy for kinase targeted therapeutics. We report the prospective discovery of ErbB2 mutations that confer resistance to the small-molecule inhibitor lapatinib. EXPERIMENTAL DESIGN: We did in vitro screening using a randomly mutagenized ErbB2 expression library in Ba/F3 cells, which were dependent on ErbB2 activity for survival and growth. RESULTS: Lapatinib resistance screens identified mutations at 16 different ErbB2 amino acid residues, with 12 mutated amino acids mapping to the kinase domain. Mutations conferring the greatest lapatinib resistance cluster in the NH2-terminal kinase lobe and hinge region. Structural computer modeling studies suggest that lapatinib resistance is caused by multiple mechanisms; including direct steric interference and restriction of conformational flexibility (the inactive state required for lapatinib binding is energetically unfavorable). ErbB2 T798I imparts the strongest lapatinib resistance effect and is analogous to the epidermal growth factor receptor T790M, ABL T315I, and cKIT T670I gatekeeper mutations that are associated with clinical drug resistance. ErbB2 mutants associated with lapatinib resistance transformed NIH-3T3 cells, including L755S and T733I mutations known to occur in human breast and gastric carcinomas, supporting a direct mechanism for lapatinib resistance in ErbB2 driven human cancers. The epidermal growth factor receptor/ErbB2/vascular endothelial growth factor receptor inhibitor EXEL-7647 was found to inhibit almost all lapatinib resistance-associated mutations. Furthermore, no ErbB2 mutations were found to be associated with EXEL-7647 resistance and lapatinib sensitivity. CONCLUSIONS: Taken together, these data suggest potential target based mechanisms of resistance to lapatinib and suggest that EXEL-7647 may be able to circumvent these effects. PMID- 18413840 TI - Therapeutic significance of elevated tissue transglutaminase expression in pancreatic cancer. AB - PURPOSE: Tissue transglutaminase (TG2) is a multifunctional protein that is implicated in development of drug resistance and metastasis. Therefore, we examined therapeutic targeting of TG2 for inhibiting growth and metastasis of in vivo growing pancreatic ductal adenocarcinoma (PDAC) in nude mice. EXPERIMENTAL DESIGN: We implanted Panc-28 pancreatic cancer cells to induce orthotopic PDAC tumors in nude mice and determined the efficacy of liposomal TG2 small interfering RNA (siRNA) either alone or in combination with gemcitabine. RESULTS: We show that down-regulation of endogenous TG2 by siRNA could effectively block the growth of PDAC. Moreover, down-regulation of TG2 significantly enhanced the therapeutic efficacy of gemcitabine against PDAC and inhibited metastatic spread of the disease. The antitumor activity was related to inhibition of proliferation, angiogenesis, and Akt phosphorylation. CONCLUSION: siRNA-mediated down-regulation of TG2 represents a promising therapeutic approach for improved treatment of PDAC. PMID- 18413841 TI - Profound inhibition of antigen-specific T-cell effector functions by dasatinib. AB - PURPOSE: The dual BCR-ABL/SRC kinase inhibitor dasatinib entered the clinic for the treatment of chronic myeloid leukemia and Ph+ acute lymphoblastic leukemia. Because SRC kinases are known to play an important role in physiologic T-cell activation, we analyzed the immunobiological effects of dasatinib on T-cell function. The effect of dasatinib on multiple T-cell effector functions was examined at clinically relevant doses (1-100 nmol/L); the promiscuous tyrosine kinase inhibitor staurosporine was used as a comparator. EXPERIMENTAL DESIGN: Purified human CD3+ cells and virus-specific CD8+ T cells from healthy blood donors were studied directly ex vivo; antigen-specific effects were confirmed in defined T-cell clones. Functional outcomes included cytokine production (interleukin-2, IFN gamma, and tumor necrosis factor alpha), degranulation (CD107a/b mobilization), activation (CD69 up-regulation), proliferation (carboxyfluorescein diacetate succinimidyl ester dilution), apoptosis/necrosis induction, and signal transduction. RESULTS: Both dasatinib and staurosporine inhibited T-cell activation, proliferation, cytokine production, and degranulation in a dose-dependent manner. Mechanistically, this was mediated by the blockade of early signal transduction events and was not due to loss of T cell viability. Overall, CD4+ T cells seemed to be more sensitive to these effects than CD8+ T cells, and naive T cells more sensitive than memory T-cell subsets. The inhibitory effects of dasatinib were so profound that all T-cell effector functions were shut down at therapeutically relevant concentrations. CONCLUSION: These findings indicate that caution is warranted with use of this drug in the clinical setting and provide a rationale to explore the potential of dasatinib as an immunosuppressant in the fields of transplantation and T-cell driven autoimmune diseases. PMID- 18413842 TI - Noninvasive imaging of apoptosis and its application in cancer therapeutics. AB - PURPOSE: Activation of the apoptotic cascade plays an important role in the response of tumors to therapy. Noninvasive imaging of apoptosis facilitates optimization of therapeutic protocols regarding dosing and schedule and enables identification of efficacious combination therapies. EXPERIMENTAL DESIGN: We describe a hybrid polypeptide that reports on caspase-3 activity in living cells and animals in a noninvasive manner. This reporter, ANLucBCLuc, constitutes a fusion of small interacting peptides, peptide A and peptide B, with the NLuc and CLuc fragments of luciferase with a caspase-3 cleavage site (DEVD) between pepANLuc (ANLuc) and pepBCLuc (BCLuc). During apoptosis, caspase-3 cleaves the reporter, enabling separation of ANLuc from BCLuc. A high-affinity interaction between peptide A and peptide B restores luciferase activity by NLuc and CLuc complementation. Using a D54 glioma model, we show the utility of the reporter in imaging of apoptosis in living subjects in response to various chemotherapy and radiotherapy regimens. RESULTS: Treatment of live cells and mice carrying D54 tumor xenografts with chemotherapeutic agents such as temozolomide and perifosine resulted in induction of bioluminescence activity, which correlated with activation of caspase-3. Treatment of mice with combination therapy of temozolomide and radiation resulted in increased bioluminescence activity over individual treatments and increased therapeutic response due to enhanced apoptosis. CONCLUSION: The data provided show the utility of the ANLucBCLuc reporter in dynamic, noninvasive imaging of apoptosis and provides a rationale for use of this technology to optimize dose and schedule of novel therapies or to develop novel combination therapies using existing drugs. PMID- 18413843 TI - Dopamine increases the efficacy of anticancer drugs in breast and colon cancer preclinical models. AB - PURPOSE: Because neurotransmitter dopamine inhibits vascular permeability factor/vascular endothelial growth factor (VEGF)-induced angiogenesis and as anti VEGF agents act synergistically with anticancer drugs, we therefore investigated whether dopamine can increase the efficacies of these drugs. EXPERIMENTAL DESIGN: The effect of dopamine was investigated in human breast cancer-(MCF-7) and colon (HT29) cancer-bearing mice. Experimental groups received either dopamine or doxorubicin or dopamine plus doxorubicin in MCF-7 tumor-bearing mice, and either dopamine or 5-fluorouracil or dopamine plus 5-fluorouracil in HT29-bearing mice. Thereafter, tumor growth, angiogenesis, tumor cell apoptosis, life span, and the effect of dopamine on the growth and survival of tumor cells in vitro were determined. Finally, the effects of dopamine on tumor vascular permeability; on VEGF receptor-2, mitogen-activated protein kinase, and focal adhesion kinase phosphorylation; and also on the proliferation and migration of tumor endothelial cells were investigated. RESULTS: Dopamine, in combination with anticancer drugs, significantly inhibited tumor growth and increased the life span when compared with treatment with dopamine or anticancer drugs alone. Dopamine had no direct effects on the growth and survival of tumor cells. The antiangiogenic action of dopamine was mediated by inhibiting proliferation and migration of tumor endothelial cells through suppression of VEGF receptor-2, mitogen-activated protein kinase, and focal adhesion kinase phosphorylation. CONCLUSION: Our study shows that dopamine significantly enhances the efficacies of commonly used anticancer drugs and also indicates that an inexpensive drug like dopamine, which is being extensively used in the clinics, might have a role as an antiangiogenic agent for the treatment of breast and colon cancer. PMID- 18413845 TI - Deaths among patients with screen-detected lung cancer. PMID- 18413846 TI - Computed tomography screening for lung cancer. PMID- 18413847 TI - Biomarkers in early clinical trials: the committed and the skeptics. PMID- 18413849 TI - T-cell response in B-cell lymphomas: favorable or unfavorable? PMID- 18413853 TI - Survival in microsatellite-unstable colorectal cancer. PMID- 18413855 TI - Population pharmacokinetc/pharmacodynamic modeling of paclitaxel and carboplatin in ovarian cancer. PMID- 18413856 TI - Effects of the beta 3-adrenergic receptor agonist disodium 5-[(2R)-2-[[(2R)-2-(3 chlorophenyl)-2-hydroxyethyl]amino]propyl]-1,3-benzodioxole-2,2-dicarboxylate (CL 316243) on bladder micturition reflex in spontaneously hypertensive rats. AB - The present study investigated whether beta3-adrenoceptor activation acts on the bladder afferent pathway by examination of the visceromotor reflex (VMR) and pressor responses to urinary bladder distension (UBD) and whether beta3 adrenoceptor activation produces urinary bladder relaxation in hyperactive spontaneously hypertensive rats (SHRs) in comparison with their normotensive control rats [Wistar-Kyoto (WKY)]. Using the VMR responses to noxious UBD as a measure of bladder afferent signal transmission, SHRs did not present a sensitized bladder phenotype. However, reduced bladder compliance accompanied by a reduced void threshold was detected in the SHR detrusor. Furthermore, the selective beta3-adrenoceptor agonist disodium 5-[(2R)-2-[[(2R)-2-(3-chlorophenyl) 2-hydroxyethyl]-amino]propyl]-1,3-benzodioxole-2,2-dicarboxylate (CL-316243) (i.v.) failed to attenuate VMR or pressor responses to UBD in either SHRs or WKY rats, but it dose-dependently inhibited rhythmic contraction (RC) in SHRs. The minimal effective dose was 0.001 mg/kg. Using the same model in WKY rats, CL 316243 did not elicit significant inhibition of contractions in the bladder RC assay. These results suggest that SHRs represent abnormal efferent/detrusor function (detrusor overactivity) without mechanosensory afferent hypersensitivity. The beta3-adrenoceptor agonist CL-316243 acts on the detrusor muscle to increase urine storage in SHRs. PMID- 18413857 TI - Hyperosmolarity-induced dilation and epithelial bioelectric responses of guinea pig trachea in vitro: role of kinase signaling. AB - Exercise-induced airway obstruction is thought to involve evaporative water loss and hyperosmolarity of the airway surface liquid. Hyperosmolar challenge of the epithelium of isolated, perfused guinea pig trachea rapidly alters transepithelial potential difference (V(t)), and it elicits smooth muscle relaxation mediated by epithelium-derived relaxing factor (EpDRF). In many cell types, protein kinases mediate responses to hyperosmolarity and regulatory volume increase. In this study, inhibitors were used to investigate the involvement of kinases and phosphatases in bioelectric responses of epithelium to hyperosmolarity and their possible relationship to EpDRF-mediated relaxation. After contraction of the perfused trachea with extraluminal methacholine, D mannitol applied intraluminally (< or = 80 mosM) increased V(t) and elicited dilation of the smooth muscle with a similar concentration-dependence; higher concentrations decreased V(t). In tracheas exposed to 30 mosM D-mannitol (approximately EC(50)), 4-(4-fluorophenyl)-2-(4-methylsulfinylphenyl)-5-(4 pyridyl)1H-imidazole (SB 203580) and SKF 86002 [6-(4-fluorophenyl)-2,3-dihydro-5 (4-pyridyl)imidazo[2,1-b]thiazole] (p38 inhibitors) potentiated the dilation, whereas SP 600125 [anthra[1,9-cd]pyrazol-6(2H)-one-1,9-pyrazoloanthrone] and dicumarol [c-Jun NH(2)-terminal kinase (JNK) inhibitors], chelerythrine [nonselective protein kinase C (PKC) inhibitor], and NaAsO(2) (mitogen-activated protein kinase stress inducer) and Na(3)VO(4) (protein tyrosine phosphatase inhibitor) inhibited the hyperpolarization. Large increases in the phosphorylation of p38 and JNK occurred at concentrations higher than those needed to elicit functional responses. The phosphatidylinositol 3-kinase inhibitor 2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one (LY 294002) and Na(3)VO(4) did not affect the V(t) responses, but they inhibited methacholine induced constriction; SP 600125 and dicumarol potentiated, and chelerythrine inhibited, methacholine-induced epithelial hyperpolarization. These results suggest that JNK, PKC, and phosphatase(s) are involved in hyperosmolarity-induced hyperpolarization of the tracheal epithelium but that p38 is involved in EpDRF mediated relaxation. PMID- 18413858 TI - Genetic deletion of BACE1 in mice affects remyelination of sciatic nerves. AB - BACE1 is a promising therapeutic and preventive target for Alzheimer's disease because it is essential for amyloid deposition. However, the recent demonstration of BACE1 in modulating developmental myelination in both peripheral and central nervous systems raises a concern of its effect on myelin maintenance or remyelination, and inhibition of these processes will potentially be detrimental to the BACE1 inhibitor users who are susceptible to myelination diseases such as adult peripheral nerve injury or multiple sclerosis. In this report, we investigated the role of BACE1 during peripheral nerve remyelination in wild-type (WT) and BACE1-null mice. We show here that genetic deletion of BACE1 affects sciatic nerve remyelination. The impaired remyelination appears to stem from the loss of neuregulin-1 cleavage by BACE1. To demonstrate a direct cleavage of neuregulin-1 by BACE1, we have identified a BACE1 cleavage site that turns out be highly conserved among neuregulin-1 paralogues. Moreover, we show that neuregulin 1 family member neuregulin-3 is also cleavable by BACE1. We hypothesize that the BACE1-cleaved extracellular domain of axonal neuregulin-1, perhaps neuregulin-3 as well, binds to Schwann cell ErbB receptors, which in turn regulate remyelination. Pharmacological inhibition of BACE1 should be carefully monitored to avoid alteration of signaling pathway that regulates remyelination. PMID- 18413859 TI - A new mechanism for the aging of hematopoietic stem cells: aging changes the clonal composition of the stem cell compartment but not individual stem cells. AB - Whether hematopoietic stem cells (HSCs) change with aging has been controversial. Previously, we showed that the HSC compartment in young mice consists of distinct subsets, each with predetermined self-renewal and differentiation behavior. Three classes of HSCs can be distinguished based on their differentiation programs: lymphoid biased, balanced, and myeloid biased. We now show that aging causes a marked shift in the representation of these HSC subsets. A clonal analysis of repopulating HSCs demonstrates that lymphoid-biased HSCs are lost and long-lived myeloid-biased HSCs accumulate in the aged. Myeloid-biased HSCs from young and aged sources behave similarly in all aspects tested. This indicates that aging does not change individual HSCs. Rather, aging changes the clonal composition of the HSC compartment. We show further that genetic factors contribute to the age related changes of the HSC subsets. In comparison with B6 mice, aged D2 mice show a more pronounced shift toward myeloid-biased HSCs with a corresponding reduction in the number of both T- and B-cell precursors. This suggests that low levels of lymphocytes in the blood can be a marker for HSC aging. The loss of lymphoid biased HSCs may contribute to the impaired immune response to infectious diseases and cancers in the aged. PMID- 18413860 TI - Structure determination of a Galectin-3-carbohydrate complex using paramagnetism based NMR constraints. AB - The determination of the location and conformation of a natural ligand bound to a protein receptor is often a first step in the rational design of molecules that can modulate receptor function. NMR observables, including NOEs, often provide the basis for these determinations. However, when ligands are carbohydrates, interactions mediated by extensive hydrogen-bonding networks often reduce or eliminate NOEs between ligand and protein protons. In these cases, it is useful to look to other distance- and orientation-dependent observables that can constrain the geometry of ligand-protein complexes. Here we illustrate the use of paramagnetism-based NMR constraints, including pseudo-contact shifts (PCS) and field-induced residual dipolar couplings (RDCs). When a paramagnetic center can be attached to the protein, field-induced RDCs and PCS reflect only bound-state properties of the ligand, even when averages over small fractions of bound states and large fractions of free states are observed. The effects can also be observed over a long range, making it possible to attach a paramagnetic center to a remote part of the protein. The system studied here is a Galectin-3-lactose complex. A lanthanide-binding peptide showing minimal flexibility with respect to the protein was integrated into the C terminus of an expression construct for the Galectin-3-carbohydrate-binding domain. Dysprosium ion, which has a large magnetic susceptibility anisotropy, was complexed to the peptide, making it possible to observe both PCSs and field-induced RDCs for the protein and the ligand. The structure determined from these constraints shows agreement with a crystal structure of a Galectin-3-N-acetyllactosamine complex. PMID- 18413861 TI - Hydrodynamic characterization of the SufBC and SufCD complexes and their interaction with fluorescent adenosine nucleotides. AB - Bacteria, as well as the plastid organelles of algae and higher plants, utilize proteins of the suf operon. These are involved in Fe-S cluster assembly, particularly under conditions of iron limitation or oxidative stress. Genetic experiments in some organisms found that the ATPase SufC is essential, though its role in Fe-S biogenesis remains unclear. To ascertain how interactions with other individual Suf proteins affect the activity of SufC we coexpressed it with either SufB or SufD from Thermotoga maritima and purified the resulting SufBC and SufCD complexes. Analytical ultracentrifuge and multiangle light-scattering measurements showed that the SufBC complex exists in solution as the tetrameric SufB(2)C(2) species, whereas SufCD exists as an equilibrium mixture of SufCD and SufC(2)D(2). Transient kinetic studies of the complexes were made using fluorescent 2'(3')-O-(N-methylanthraniloyl-(mant) analogues of ATP and ADP. Both SufBC and SufCD bound mantATP and mantADP much more tightly than does SufC alone. Compared to the cleavage step of the mantATPase of SufC alone, that of SufBC was accelerated 180-fold and that of SufCD only fivefold. Given that SufB and SufD have 20% sequence identity and similar predicted secondary structures, the different hydrodynamic properties and kinetic mechanisms of the two complexes are discussed. PMID- 18413862 TI - Alternative stabilities of a proline-rich antibacterial peptide in vitro and in vivo. AB - The proline-rich designer antibacterial peptide dimer A3-APO is currently under preclinical development for the treatment of systemic infections caused by antibiotic-resistant Gram-negative bacteria. The peptide showed remarkable stability in 25% mouse serum in vitro, exhibiting a half-life of approximately 100 min as documented by reversed-phase chromatography. Indeed, after a 30-min incubation period in undiluted mouse serum ex vivo, mass spectrometry failed to identify any degradation product. The peptide was still a major peak in full blood ex vivo, however, with degradation products present corresponding to amino terminal cleavage. When injected into mice intravenously, very little, if any unmodified peptide could be detected after 30 min. Nevertheless, the major early metabolite, a full single-chain fragment, was detectable until 90 min, and this fragment exhibited equal or slightly better activity in the broth microdilution antimicrobial assay against a panel of resistant Enterobactericeae strains. The Chex1-Arg20 metabolite, when administered three times at 20 mg/kg to mice infected with a sublethal dose (over LD(50)) of an extended spectrum beta lactamase-producing Escherichia coli strain, completely sterilized the mouse blood, similar to imipenem added at a higher dose. The longer and presumably more immunogenic prodrug A3-APO, injected subcutaneously twice over a 3-wk period, did not induce any antibody production, indicating the suitability of this peptide or its active metabolite for clinical development. PMID- 18413863 TI - The cover. Disks of Newton (study for "fugue in two colors"). PMID- 18413864 TI - A piece of my mind. Rediscovery. PMID- 18413865 TI - Critics say FDA's off-label guidance allows marketing disguised as science. PMID- 18413866 TI - Group seeks to improve nonmedical aspects of health in the United States. PMID- 18413867 TI - FDA to boost its presence in China. PMID- 18413868 TI - Guideline: dementia drugs' benefits uncertain. PMID- 18413869 TI - Prognosis of transient neurological attacks. PMID- 18413870 TI - Prognosis of transient neurological attacks. PMID- 18413871 TI - Demographic characteristics and opioid prescribing. PMID- 18413872 TI - Demographic characteristics and opioid prescribing. PMID- 18413873 TI - Determining hospice benefit for patients with dementia. PMID- 18413874 TI - Guest authorship and ghostwriting in publications related to rofecoxib: a case study of industry documents from rofecoxib litigation. AB - CONTEXT: Authorship in biomedical publication provides recognition and establishes accountability and responsibility. Recent litigation related to rofecoxib provided a unique opportunity to examine guest authorship and ghostwriting, practices that have been suspected in biomedical publication but for which there is little documentation. OBJECTIVE: To characterize different types and the extent of guest authorship and ghostwriting in 1 case study. DATA SOURCES: Court documents originally obtained during litigation related to rofecoxib against Merck & Co Inc. Documents were created predominantly between 1996 and 2004. In addition, publicly available articles related to rofecoxib identified via MEDLINE. DATA EXTRACTION: All documents were reviewed by one author, with selected review by coauthors, using an iterative process of review, discussion, and rereview of documents to identify information related to guest authorship or ghostwriting. DATA SYNTHESIS: Approximately 250 documents were relevant to our review. For the publication of clinical trials, documents were found describing Merck employees working either independently or in collaboration with medical publishing companies to prepare manuscripts and subsequently recruiting external, academically affiliated investigators to be authors. Recruited authors were frequently placed in the first and second positions of the authorship list. For the publication of scientific review papers, documents were found describing Merck marketing employees developing plans for manuscripts, contracting with medical publishing companies to ghostwrite manuscripts, and recruiting external, academically affiliated investigators to be authors. Recruited authors were commonly the sole author on the manuscript and offered honoraria for their participation. Among 96 relevant published articles, we found that 92% (22 of 24) of clinical trial articles published a disclosure of Merck's financial support, but only 50% (36 of 72) of review articles published either a disclosure of Merck sponsorship or a disclosure of whether the author had received any financial compensation from the company. CONCLUSIONS: This case study review of industry documents demonstrates that clinical trial manuscripts related to rofecoxib were authored by sponsor employees but often attributed first authorship to academically affiliated investigators who did not always disclose industry financial support. Review manuscripts were often prepared by unacknowledged authors and subsequently attributed authorship to academically affiliated investigators who often did not disclose industry financial support. PMID- 18413875 TI - Reporting mortality findings in trials of rofecoxib for Alzheimer disease or cognitive impairment: a case study based on documents from rofecoxib litigation. AB - Sponsors have a marketing interest to represent their products in the best light. This approach conflicts with scientific standards that require the symmetric and comparable reporting of safety and efficacy data. Selective reporting of the results of clinical trials can misrepresent the risk-benefit profile of drugs. We summarize how the sponsor represented mortality findings associated with rofecoxib in clinical trials of patients with Alzheimer disease or cognitive impairment. We reviewed documents that became available during litigation related to rofecoxib involving Merck & Co, including internal company analyses and information provided by the sponsor to the FDA. We also evaluated information in 2 published articles that reported results of these trials. In one article (reporting results of protocol 091) published in 2004, 11 "non-drug related deaths" were reported (9 deaths among 346 rofecoxib patients and 2 deaths among 346 placebo patients). In another article (reporting results of protocol 078) published in 2005, 39 deaths were reported among patients taking study treatment or within 14 days of the last dose (24 among 725 rofecoxib patients and 15 among 732 placebo patients) and an additional 22 deaths in the off-drug period (17 in rofecoxib patients and 5 in placebo patients). However, these articles did not include analyses or statistical tests of the mortality data, and the 2 articles concluded that regarding safety, rofecoxib is "well tolerated." In contrast, in April 2001, the company's internal intention-to-treat analyses of pooled data from these 2 trials identified a significant increase in total mortality (hazard ratio [HR], 4.43; 95% CI, 1.26-15.53 for protocol 091, and HR, 2.55; 95% CI, 1.17 5.56 for protocol 078), with overall mortality of 34 deaths among 1069 rofecoxib patients and 12 deaths among 1078 placebo patients (HR, 2.99; 95% CI, 1.55-5.77). These mortality analyses were neither provided to the FDA nor made public in a timely fashion. The data submitted by the sponsor to the FDA in a Safety Update Report in July 2001 used on-treatment analysis methods and reported 29 deaths (2.7%) among 1067 rofecoxib patients and 17 deaths (1.6%) among 1075 placebo patients. This on-treatment approach to reporting minimized the appearance of any mortality risk. In December 2001, when the FDA raised safety questions about the submitted safety data, the sponsor did not bring these issues to an institutional review board for review and revealed that there was no data and safety monitoring board for the protocol 078 study. The findings from this case study suggest that additional protections for human research participants, including new approaches for the conduct, oversight, and reporting of industry-sponsored trials, are necessary. PMID- 18413876 TI - Palliative care for patients with head and neck cancer: "I would like a quick return to a normal lifestyle". AB - Head and neck cancers constitute a diverse group of diseases including malignancies of the oral cavity, oropharynx, larynx, sinuses, and skull base. Treatment of these cancers includes a combination of surgical resection, chemotherapy, and radiation. Due to both the patterns of disease recurrence and the adverse effects of treatments, patients with head and neck cancer often have a complex and prolonged course of illness that is marked by periods of freedom from disease and symptoms interspersed with bouts of serious illness, debility, and numerous physical and psychological symptoms including pain, dysphagia, weight loss, disfigurement, depression, and xerostomia. Thus, management of this disease is best provided by an interdisciplinary team that includes individuals from the disciplines of otolaryngology, palliative care, radiation oncology, oncology, nutrition, speech, and physical and occupational therapy. Using the case of Mr K, we describe the symptoms encountered by patients with head and neck cancer and suggest options for management. We discuss the psychological aspects that affect these patients, including issues such as changes in body image, quality of life, anxiety, and guilt. Finally, we discuss the importance of the interdisciplinary team in the care of these patients and outline the roles of each team member. By providing comprehensive care to patients with malignancies of the head and neck, clinicians can increase the likelihood that patients and their families will be able to obtain the best possible outcomes and quality of life. PMID- 18413877 TI - Management of intractable nausea and vomiting in patients at the end of life: "I was feeling nauseous all of the time . . . nothing was working". PMID- 18413878 TI - The international migration and recruitment of nurses: human rights and global justice. PMID- 18413879 TI - Gizmo idolatry. PMID- 18413880 TI - Impugning the integrity of medical science: the adverse effects of industry influence. PMID- 18413881 TI - JAMA patient page. Coronary artery bypass grafting. PMID- 18413882 TI - GABA effects on neurogenesis: an arsenal of regulation. AB - The well-known inhibitory neurotransmitter GABA is excitatory to immature neurons during development as well as to neural progenitor cells during adult neurogenesis. Recent evidence indicates that GABA plays a role in regulating neurogenesis in various situations. This journal club paper aims to provide a skeleton description of GABA regulation of neurogenesis, describing some of the disparate mechanisms and scenarios in which it is involved. PMID- 18413883 TI - Eph, a protein family coming of age: more confusion, insight, or complexity? AB - Since the mid-1980s, Eph receptors have evolved from being regarded as orphan receptors with unknown functions and ligands to becoming one of the most complex "global positioning systems" that regulates cell traffic in multicellular organisms. During this time, there has been an exponentially growing interest in Ephs and ephrin ligands, coinciding with important advances in the way biological function is interrogated through mapping of genomes and manipulation of genes. As a result, many of the original concepts that used to define Eph signaling and function went overboard. Clearly, the need for progress in understanding Eph ephrin biology and the underlying molecular principles involved has been compelling. Many cell-positioning programs during normal and oncogenic development-in particular, the patterning of skeletal, vascular, and nervous systems-are modulated in some way by Eph-ephrin function. Undeniably, the complexity of the underlying signaling networks is considerable, and it seems probable that systems biology approaches are required to further improve our understanding of Eph function. PMID- 18413884 TI - In vivo near-infrared fluorescence imaging of carcinoembryonic antigen-expressing tumor cells in mice. AB - PURPOSE: To prospectively depict carcinoembryonic antigen (CEA)-expressing tumors in mice with a high-affinity probe consisting of a near-infrared (NIR) fluorochrome and the clinically used anti-CEA antibody fragment arcitumomab. MATERIALS AND METHODS: This study was approved by the regional animal committee. By coupling a NIR fluorescent (NIRF) cyanine dye (DY-676) to a specific antibody fragment directed against CEA (arcitumomab) and a nonspecific IgG Fab fragment, a bio-optical high-affinity fluorescent probe (anti-CEA-DY-676) and a low-affinity fluorescent probe (FabIgG-DY-676) were designed. The dye-to-protein ratios were determined, and both probes were tested for NIRF imaging in vitro on CEA expressing LS-174T human colonic adenocarcinoma cells and CEA-nonexpressing A-375 human melanoma cells by using a bio-optical NIR small-animal imager. In vivo data of xenografted LS-174T and A-375 tumors in mice (n = 10) were recorded and statistically analyzed (Student t test). RESULTS: The dye-to-protein ratios were determined as 3.0-3.5 for both probes. In vitro experiments revealed the specific binding of the anti-CEA-DY-676 probe on CEA-expressing cells as compared with CEA nonexpressing cells; the FabIgG-DY-676 probe showed a markedly lower binding affinity to cells. In vivo LS-174T tumors xenografted in all mice could be significantly distinguished from A-375 tumors with application of the anti-CEA-DY 676 but not with that of the FabIgG-DY-676 at different times (2-24 hours, P < .005) after intravenous injection of the probes. Semiquantitative analysis revealed maximal fluorescence signals of anti-CEA-DY-676 to CEA-expressing tumors about 8 hours after injection. CONCLUSION: Findings of this study indicate the potential use of the high-affinity probe anti-CEA-DY-676 for specific NIRF imaging in in vivo tumor diagnosis. PMID- 18413885 TI - Management of small (3-5-mm) pulmonary nodules at chest CT: global survey of thoracic radiologists. AB - PURPOSE: To prospectively determine management strategies used by international thoracic radiologists in evaluation of small (3-5-mm) pulmonary nodules at chest computed tomography (CT). MATERIALS AND METHODS: Institutional review board exemption was granted for this study, which included consenting participants. An electronic survey was sent to members of major thoracic radiology societies in North America, Europe, and Asia. The main part of the survey consisted of four management questions with clinical scenarios. Associations between recommendations and years of experience, location in a region endemic for granulomatous disease, country, and practice type were assessed. Univariate analysis was performed to determine differences in follow-up recommendations on the basis of patient characteristics, percentage of chest CT scans obtained at follow-up, years of experience in radiology, and professional society affiliation of respondents. Differences in categorical variables were examined by using Pearson chi(2) and Fisher exact tests. RESULTS: Two hundred two (25%) of approximately 800 online surveys were completed. Surveys from respondents from the United States comprised 61% of completed surveys. Median experience of respondents was 11-20 years. Fifty-two percent practice in an area endemic for granulomatous disease. Only 35% of practices have a policy in place for nodule management. In scenarios in which patients had a low likelihood of malignancy, respondents' preferential recommendation was short-term CT follow-up (3-6 months) rather than intermediate-term CT follow-up (12 months) for patients older than 40 years compared with their recommendation in patients younger than 40 years, in whom recommendations for short- or intermediate-term follow-up were roughly equal. In scenarios in which patients had a high risk of malignancy, follow-up was also strongly favored instead of biopsy, with short-term follow-up more commonly advocated. Location in an area endemic for granulomatous disease and years of experience also influenced recommendations. CONCLUSION: Globally, the most common recommendation for CT evaluation of nodules is short-term follow-up, with a tendency toward less aggressiveness in scenarios in which patients had lower risk of malignancy and increased aggressiveness in scenarios in which patients had higher risk of malignancy. PMID- 18413886 TI - Pixel distribution analysis: can it be used to distinguish clear cell carcinomas from angiomyolipomas with minimal fat? AB - PURPOSE: To retrospectively determine if pixel histogram analysis of unenhanced computed tomographic (CT) images can be used to distinguish angiomyolipomas (AMLs) with minimal fat from clear cell renal cell carcinomas (CCRCCs). MATERIALS AND METHODS: The human studies committee approved this HIPAA-complaint study, with waiver of informed consent. Patients with pathologically proved AMLs lacking visible macroscopic fat at CT and patients with pathologically proved CCRCCs were included. Lesions were measured, and a histogram (number of pixels with each attenuation) was calculated electronically within a central region of interest. The percentage of pixels below the attenuation thresholds -20 HU and 10 HU was calculated in both cohorts. The unpaired Student t test was used to compare the average percentage of subthreshold pixels at each threshold. P < .05 indicated a significant difference. The number of lesions with more than the selected percentage of subthreshold pixels was calculated in both groups, and the chi(2) test was used to test the significance of differences between cohorts. The area under the receiver operating characteristic (ROC) curve was used to determine if any percentage of subthreshold pixels could be used to differentiate between the two cohorts. RESULTS: There were 22 patients with pathologically proved AMLs lacking visible macroscopic fat on CT images. Tuberous sclerosis affected three of these patients. Mean maximal transverse lesion diameter was 20 mm (range, 11 38 mm). There were 28 patients in the CCRCC comparison group. Mean maximal transverse lesion diameter was 26 mm (range, 15-36 mm). Neither the Student t test (P > .2 for all thresholds <0 HU) nor the chi(2) test (P > .15 for all thresholds <0 HU) revealed a significant difference between cohorts. A lesion with more low-attenuation pixels was significantly more likely to be characterized as CCRCC than as AML with ROC curve analysis. CONCLUSION: Once AMLs with visible fat on CT images are excluded, pixel histogram analysis cannot be used to distinguish between AMLs and CCRCCs. PMID- 18413887 TI - Opacification of urinary bladder and ureter at CT urography: effect of a log rolling procedure and postvoiding residual bladder urine volume. AB - PURPOSE: To retrospectively evaluate the effect of a log-rolling procedure and postvoiding residual (PVR) bladder urine volume on opacification of urinary bladder and ureters at multi-detector row computed tomographic (CT) urography. MATERIALS AND METHODS: Institutional review board approval was obtained, and informed consent was waived for this retrospective HIPAA-compliant study. Triple phase 16- or 64-detector row CT urographic images in 166 patients (88 men, 78 women; mean age, 58.9 years; range, 22-89 years) were evaluated retrospectively. Immediately prior to excretory phase scanning, 67 patients did and 99 did not undergo a log-rolling procedure on the CT table. PVR bladder urine volume was quantified as the largest cross-sectional area of the bladder measured on unenhanced images (PVR values). The degree of bladder opacification was quantified as the percentage of the total cross-sectional area of the bladder that was opacified on excretory phase images. Ureteral opacification was quantified as the percentage of ureteral length that contained enhanced urine. On the basis of PVR values, patients were stratified into four subgroups (2000 to 3000 to 4000 mm(2)). The Wilcoxon rank sum and Student t tests were used to evaluate differences. RESULTS: Median degree of bladder opacification of the log-rolling versus non-log-rolling group was 100% versus 78% for PVR values of 2000 mm(2) or less (P < .01), 99% versus 79% for PVR values of more than 2000 to 3000 mm(2) or less (P = .01), 89% versus 79% for PVR values of more than 3000 to 4000 mm(2) or less (P < .05), and 64% versus 69% for PVR values of more than 4000 mm(2) (P = .96). There was no significant difference between ureteral opacification and log rolling or between bladder and ureteral opacification (P > .05). CONCLUSION: Use of a log-rolling procedure prior to excretory phase CT urography increases the percentage of bladder opacification in patients with PVR values of 4000 mm(2) or less. No difference in ureteral opacification was observed between the log-rolling and non-log-rolling groups. PMID- 18413888 TI - Canine prostate: contrast-enhanced US-guided radiofrequency ablation with urethral and neurovascular cooling--initial experience. AB - PURPOSE: To prospectively evaluate in a canine model contrast material-enhanced ultrasonography (US) for guiding and monitoring radiofrequency (RF) ablation of the entire prostate, with urethral and vascular cooling to protect the surrounding structures. MATERIALS AND METHODS: After approval by the institutional animal use and care committee, an RF electrode was used to ablate the entire prostate in 15 dogs. During ablation, pulse-inversion harmonic US was performed by using an endocavitary probe after an intravenous bolus injection (0.04 mL/kg) and infusion (0.015 muL/kg/min) of a US contrast agent. In group 1 (n = 4), no cooling protection was used during ablation. In group 2 (n = 5), urethral and bladder protection was provided by inserting a 12-F catheter infused with cold saline (8 degrees C +/- 4 [standard deviation]) at a rate of 100 mL/min. In group 3 (n = 6), further protection of the neurovascular bundles (NVBs) was provided by infusing cold saline (8 degrees C +/- 4) into the iliac arteries at a rate of 50 mL/min by means of catheterization of the femoral artery. Pathologic findings among the three groups were compared by using the Wilcoxon rank sum test. RESULTS: The average volumes of prostate ablation achieved in the three groups were 96.6%, 91.9%, and 92%. Contrast-enhanced pulse inversion harmonic US allowed visualization and monitoring of urethral and NVB blood flow during the ablation. Without protection, damage to the urethra and the NVB was demonstrated at both US and pathologic examination. There was highly significant difference in urethral damage between groups with and the group without urethral cooling (P = .002), while intraarterial cooling demonstrated a nonsignificant trend toward a decreased NVB damage (P = .069). CONCLUSION: Contrast-enhanced US can guide RF ablation of the entire prostate. Infusion of cold saline provides effective protection for the urethra during such procedures. The application of intraarterial cooling did not provide a significant improvement in the protection of the NVB in this small study. PMID- 18413889 TI - Coronary calcium coverage score: determination, correlates, and predictive accuracy in the Multi-Ethnic Study of Atherosclerosis. AB - PURPOSE: To develop a new calcium score for use with unenhanced cardiac computed tomography (CT) that can be used to define the percentage of coronary arteries affected by calcium and to correlate this score with risk factors and cardiovascular events. MATERIALS AND METHODS: Institutional review boards at all participating centers approved this HIPAA-compliant study, and all participants gave written informed consent. Calcium coverage score (CCS), which represents the percentage of coronary arteries affected by calcific plaque, was calculated for 3252 participants in the Multi-Ethnic Study of Atherosclerosis in whom calcific plaque was detected with CT. Quasi-Poisson models were used to estimate associations (assessed by using t tests with robust standard errors) between CCS and risk factors. Associations between the CCS, Agatston, and calcium mass scores (hereafter, mass scores) and outcomes were estimated and assessed by using Cox proportional hazards models with Wald tests. The predictive ability of these models was assessed by using area under the receiver operating characteristic curves and bootstrap t tests. RESULTS: After adjustments were made for age, race, ethnicity, and sex in the quasi-Poisson model, CCS was associated with hypertension, dyslipidemia, and diabetes (P < .001 for all diseases). After adjustments for age and sex, a twofold increase in CCS was associated with a 52% (95% confidence interval: 34%, 72%) increase in risk for any coronary heart disease (CHD) event. When Agatston or mass scores were included with CCS in a Cox model for prediction of CHD events, neither Agatston score nor mass score was a significant predictor, whereas CCS remained significantly associated with CHD events. Although receiver operating characteristic curves suggested that there was a difference between CCS score and Agatston and mass scores in prediction of a cardiac event, no differences in prediction of hard cardiac events (myocardial infarction, death) were found. CONCLUSION: Both spatial distribution and amount of calcified plaque contribute to risk for CHD. PMID- 18413890 TI - Onset of cardiac iron loading in pediatric patients with thalassemia major. AB - We reviewed cardiac T2* assessments from 77 thalassemia major patients between the ages of 2.5 and 18 years to study optimal timing of cardiac iron screening by magnetic resonance imaging. No patient under 9.5 years of age showed detectable cardiac iron in contrast to 36% of patients between the ages of 15-18 years old, corresponding to an odds-ratio of 1.28 (28%) per year. All patients with cardiac iron had received at least 35 grams of transfusional iron. Liver iron and ferritin failed to predict cardiac iron loading. Initiation of cardiac magnetic resonance imaging assessment should be determined according to age and transfusional burden rather than indices of iron overload. When appropriate chelation therapy has been administered since birth, cardiac magnetic resonance imaging can be postponed until 8 years of age when anesthesia is not required. Patients with suboptimal chelation, increased transfusional requirements, or who have initiated transfusions later in life should be tested sooner. PMID- 18413891 TI - Italian Society of Hematology practice guidelines for the management of iron overload in thalassemia major and related disorders. AB - New measures of iron accumulation in liver and heart (superconducting quantum inference device and magnetic resonance imaging), and oral iron chelators (deferiprone and deferasirox) are available for managing iron overload in thalassemia major. To assure appropriate use of these new health technologies, the Italian Society of Hematology appointed a panel of experts to produce clinical practice-guidelines for the management of iron overload in thalassemia major and related disorders. The analytical hierarchy process, a technique for multicriteria decision analysis, was applied to relevant key questions in order to identify the alternative strategies, generate explicit criteria for their evaluation, and check how well the alternatives fulfilled the criteria. The result of a comprehensive systematic review of articles released from 1990 to 2007 was used as a source of scientific evidence to compare the decisional options pairwise, and select the final recommendation. Every step in the model was developed from questionnaires and group discussion. The resulting recommendations advise about which examination to carry out in order to plan iron chelation therapy, when to start iron chelation, which iron chelator to choose in regularly transfused patients, how to monitor iron chelation therapy, and when and how to switch standard therapy. PMID- 18413892 TI - Eligibility for allogeneic transplantation in very high risk childhood acute lymphoblastic leukemia: the impact of the waiting time. AB - The advantage of allogeneic transplant from compatible related donors versus chemotherapy in children with very-high-risk acute lymphoblastic leukemia in first complete remission was previously demonstrated in an international prospective trial. This study quantified the impact of time elapsed in first remission in the same cohort. Of 357 pediatric patients with very-high-risk acute lymphoblastic leukemia, 259 received chemotherapy, 55 transplantation from compatible related and 43 from unrelated donors. The 5-year disease-free survival was 44.2% overall and 42.5% for chemotherapy only patients. The chemotherapy conditional 5-year disease-free survival increased to 44.4%, 47.6%, 51.7%, and 60.4% in patients who maintained their first remission for at least 3, 6, 9, and 12 months respectively. The overall outcome was superior to that obtained with chemotherapy-only at any time-point. The relative advantage of transplant from compatible related donors in very-high-risk childhood acute lymphoblastic leukemia was consistent for any time elapsed in first remission. PMID- 18413893 TI - {beta}-thalassemia major evolution from {beta}-thalassemia minor is associated with paternal uniparental isodisomy of chromosome 11p15. AB - beta-thalassemia major can be caused by homozygosity or compound heterozygosity for beta-globin gene mutations (HBB gene). Most cases are inherited from parents who both have diseased alleles of the HBB gene. We report a patient with late onset beta-thalassemia major that evolved from beta-thalassemia minor in which only one of her parents had the diseased HBB gene. To study the cause of beta thalassemia major in this patient, we performed the 100K single nucleotide polymorphism genotyping assay, fluorescence in situ hybridization, and DNA methylation analysis of the imprinting genes near the HBB gene. The results showed a loss of heterozygosity in the region of chromosome 11p14.3 to 11p15.5, which perfectly matched one allele of her father. Our study demonstrates that paternal uniparental isodisomy of chromosomal 11p15.5 is associated with the beta thalassemia major in this patient. Key words: beta-thalassemia major, uniparental isodisomy, mosaicism. PMID- 18413894 TI - Culture and context: East Asian American and European American differences in P3 event-related potentials and self-construal. AB - Research has demonstrated differences in social and cognitive processes between East Asians and European Americans. Whereas East Asians have been characterized as being more sensitive to situational context and attending more to the perceptual field, European Americans have been characterized as being more focused on the object and being more field independent. The goal of the present experiment was to investigate differences in neural responses to target objects and stimulus context between East Asian Americans and European Americans using a three-stimulus novelty P3 event-related potential design. As hypothesized, European Americans displayed relatively greater target P3 amplitudes, indexing attention to target events, whereas East Asian Americans displayed relatively greater novelty P3 amplitudes, indexing attention to contextually deviant events. Furthermore, the authors found that interdependent self-construal mediated the relationship between culture and the novelty P3. These findings identify a specific pattern of neural activity associated with established cultural differences in contextual sensitivity. PMID- 18413895 TI - A method with flexible and balanced control of false negatives and false positives for hit selection in RNA interference high-throughput screening assays: a statistical terminology. AB - Zhang suggests a new method that is flexible and controls the balance between false negatives and false positives for hit selection in RNA high-throughput screening assays. The author shows that the same decision rules and balances can be expressed by familiar statistical terms such as type I error and power and hence connects the new method to known statistical tools. (Journal of Biomolecular Screening 2008:309-311). PMID- 18413897 TI - Intracoronary abciximab use in patients undergoing PCI at a community hospital: a single operator experience. AB - OBJECTIVE: To evaluate the safety of intracoronary (IC) abciximab during percutaneous coronary intervention (PCI). BACKGROUND: Adjunctive treatment with glycoprotein IIb/IIIa inhibitors, especially abciximab, during PCI has been shown to improve clinical and procedural outcomes in numerous studies. However, significant bleeding complications exist with its use and this has limited its standard use. Interest has grown in local (IC) use with studies showing safety and long-term effectiveness, especially in patients with high thrombus loads. METHODS: A retrospective review of records in a database of patients who had PCI by a single operator at the Easton Hospital. RESULTS: 611 patients received IC abciximab, and there were no complications in 610 (98.3%) patients; only 1 had an allergic reaction. CONCLUSIONS: IC abciximab is safe and has a unique role in the catheterization lab and in patients at high risk of bleeding complications who would benefit from its limited use. PMID- 18413898 TI - The protective effect of bergamot oil extract on lecitine-like oxyLDL receptor-1 expression in balloon injury-related neointima formation. AB - Lectin-like oxyLDL receptor-1 (LOX-1) has recently been suggested to be involved in smooth muscle cell (SMC) proliferation and neointima formation in injured blood vessels. This study evaluates the effect of the nonvolatile fraction (NVF), the antioxidant component of bergamot essential oil (BEO), on LOX-1 expression and free radical generation in a model of rat angioplasty. Common carotid arteries injured by balloon angioplasty were removed after 14 days for histopathological, biochemical, and immunohistochemical studies. Balloon injury led to a significant restenosis with SMC proliferation and neointima formation, accompanied by increased expression of LOX-1 receptor, malondialdehyde and superoxide formation, and nitrotyrosine staining. Pretreatment of rats with BEO NVF reduced the neointima proliferation together with free radical formation and LOX-1 expression in a dose-dependent manner. These results suggest that natural antioxidants may be relevant in the treatment of vascular disorders in which proliferation of SMCs and oxyLDL-related endothelial cell dysfunction are involved. PMID- 18413899 TI - Wolman disease/cholesteryl ester storage disease: efficacy of plant-produced human lysosomal acid lipase in mice. AB - Lysosomal acid lipase (LAL) is an essential enzyme that hydrolyzes triglycerides (TGs) and cholesteryl esters (CEs) in lysosomes. Genetic LAL mutations lead to Wolman disease (WD) and cholesteryl ester storage disease (CESD). An LAL-null (lal(-/-)) mouse model resembles human WD/CESD with storage of CEs and TGs in multiple organs. Human LAL (hLAL) was expressed in Nicotiana benthamiana using the GENEWARE expression system (G-hLAL). Purified G-hLAL showed mannose receptor dependent uptake into macrophage cell lines (J774E). Intraperitoneal injection of G-hLAL produced peak activities in plasma at 60 min and in the liver and spleen at 240 min. The t(1/2) values were: approximately 90 min (plasma), approximately 14 h (liver), and approximately 32 h (spleen), with return to baseline by approximately 150 h in liver and approximately 200 h in spleen. Ten injections of G-hLAL (every 3 days) into lal(-/-) mice produced normalization of hepatic color, decreases in hepatic cholesterol and TG contents, and diminished foamy macrophages in liver, spleen, and intestinal villi. All injected lal(-/-) mice developed anti-hLAL protein antibodies, but suffered no adverse events. These studies demonstrate the feasibility of using plant-expressed, recombinant hLAL for the enzyme therapy of human WD/CESD with general implications for other lysosomal storage diseases. PMID- 18413900 TI - Effect of age and height on trunk sway during stance and gait. AB - The aim of the studies reported here was to quantify changes in balance control for stance and gait tasks with age and to pinpoint possible advantages and difficulties in using these tasks and measures derived from them to identify pathological balance control in patients. Some 470 normal subjects in the age range 6 to 82 were examined for a battery of 14 stance and gait tasks. During the tasks, angular velocity transducers mounted at lumbar 1-3 measured pitch and roll angular velocities of the body. A combination of outcome measures from several tasks was used to create an overall balance control index. Three types of sensory analyses on pitch angle and velocity amplitudes for stance trials were used to quantify possible changes in the contributions of visual, somatosensory and vestibular inputs to balance control with age for 2-legged stance tasks. Correlation analysis on task variables was used to determine the relationship of subjects' age and height on outcome measures. Outcome measures showed a characteristic "L" or "U" shaped profile with a rapid decrease in values between 7 and 25 years of age, a plateau until 55 then a gradual increase with age after 55 years of age for most stance and gait tasks. The sensory analysis technique using differences between stance tests indicated that visual contributions to balance control continuously increased with age between the ages of 15 and 80, and vestibular and lower leg somatosensory contributions remain relatively constant with age. Sensory analysis calculated as commonly-used quotients of outcome measures revealed large variance across all ages, asymmetric distributions, and no clear trends in sensory contributions to stance with age. A third technique based on a discriminant function analysis using measures from model patient populations indicated that proprioceptive but not vestibular contributions first increased with age and then decreased after 55 years of age. Correlations of outcome measures with age and height indicated that both contributed equally to changes in outcome measures between the ages of 7 and 25, otherwise height had no effect. We conclude that both stance and gait tasks should be selected for identifying changes in balance control from that of healthy persons with a preference for gait tasks as these show less variation with age. Because of the large increases in variance in the elderly and those younger than 20 years, appropriate age-matched reference values should be employed to ascertain if trunk sway is out of normal ranges. PMID- 18413901 TI - Presbyvertigo-aging of otoconia and vestibular sensory cells. AB - Imbalance is a common clinical problem in elderly persons. Subsequently falls and fractures may occur. Age-related balance problems constitute an underestimated but prominent public health problem and a socioeconomic burden. At the histological level the peripheral vestibular system in animals and humans exhibits a variety of age-related changes. The number of otoconia in the utricule and the saccule is reduced with increasing age. Degenerated otoconia caused by aging showed distinct changes of the shape. Moreover age-related decreasing hair cell counts, lipofuscin inclusions and deformation of cilia are observed in human vestibular sensory and supporting cells. Results of vestibular function testing of age-related balance disorders still remain controversially. Regular balance training and vestibular rehabilitation results in less more falls and imbalance. PMID- 18413902 TI - Characterization of age-related changes in vestibular evoked myogenic potentials. AB - A tone-burst stimulation of 500 Hz seems to be clinically most appropriate to elicit vestibular evoked myogenic potentials (VEMPs) because those VEMPs can be recorded at the lowest stimulus intensity possible. However, little is known about gender and age-related changes of the amplitude in tone-burst (500 Hz) evoked VEMPs. The aim of the present paper was therefore to investigate the influence of gender and age on VEMP amplitude in relation to the tonic muscle activity. VEMPs of 64 healthy subjects were recorded ipsilaterally during air- or bone-conducted tone burst stimulation. The EMG of the tonically activated sternocleidomastoid muscle was recorded ipsilaterally with surface electrodes. Averages were taken for P1/N1 amplitudes of male and female volunteers within 3 different age groups. Although the amplitude decreased with increasing age the tonic activity was not significant different between the age groups. Consequently the relation between VEMP amplitude and tonic muscle activity decreased with increasing age. The normative values of the age-dependent relation between VEMP amplitude and tonic muscle activity were described by the 90% confidence interval of the individual values. Normative thresholds were calculated. Normal saccular receptor function could be diagnosed if the VEMP amplitude is above (or equal to) the normative value at a given tonic muscle activity and age. Normative data as described above are required to diagnose isolated saccular defects, which are indicative of a vestibular disorder. PMID- 18413903 TI - Ocular torsion response to active head-roll movement under one-g and zero-g conditions. AB - Transitions to and from microgravity, as experienced during a spaceflight mission, radically alter the demands on sensorimotor coordination. In this contribution, attention is directed to the vestibulo-oculomotor response to active head roll-tilt, generally referred to as ocular counterroll (OCR). Results are presented from a single-case longitudinal study over a 435-day spaceflight and from three further subjects over a 30-day period in microgravity. 1. Under one-g test conditions, with the head initially in the comfortable-upright position, active head-to-trunk roll tilt elicits a combined canal- and otolith mediated oculomotor response, which manifests as a volley of torsional nystagmus beats combined with a tonic OCR. In microgravity it appears that only the transitory canal-mediated torsional nystagmus response remains. In both conditions the initial nystagmus response commences with an anticompensatory torsional fast phase. 2. Under zero-g conditions the head movements were comparable to those under one-g conditions but a consistent reduction in head velocity was observed. Despite this, eye velocity and eye-head velocity gain for the torsional component were found to be enhanced by up to 50% over the first thirty days in prolonged microgravity. 3. The results obtained from the 435-day mission indicate that the initially enhanced response decreases--over the course of several months--to preflight baseline level. The findings indicate that otolith- and canal-ocular responses are not simply added linearly, but rather that the afferent otolith signal also plays an inhibitory, or stabilising role on the canal-mediated response. Further, presuming a re-weighting of otolithic afferent information during prolonged microgravity, it is proposed that a corollary inverse re-weighting of corollary neck-proprioceptive afferences provides an effective substitute. In contrast to the idea that the torsional VOR is an evolutionary relic, it is postulated from the above findings that the anticompensatory saccade and the inherent low gain of OCR result as a compromise between intended reorientation to a tilted visual field and VOR compensation. PMID- 18413904 TI - Otolith-ocular responses in Meniere's patients before and after endolymphatic shunt operation. AB - Caloric testing in prone/supine position and constant velocity off-vertical axis rotation (OVAR) in yaw axis (rotate-then-tilt paradigm) can evaluate labyrinth function and vestibular-ocular reflex (VOR) behaviour before and after endolymphatic shunt surgery (ESS). Preoperative and postoperative otolith dysfunction can be documented by constant velocity OVAR, before the VOR is modulated by the vestibular compensation. Vestibulo-ocular responses in prone/supine position and linear VOR (lVOR) OVAR responses were observed in 10 patients before and after ESS. Ipsilateral caloric reaction in prone/supine position was reduced after ESS. Otolith-ocular function and canal-otolith interaction were improved postoperatively. Meniere's patients with bias component opposite to normal when rotating towards the lesioned ear showed relief of symptoms postoperatively. The bias component returning to normal can help to identify the relief of Meniere's attacks after ESS. The canal-otolith interaction can be observed pre- and postoperatively by means of caloric reaction in prone/supine position as part of the clinical routine. PMID- 18413905 TI - Efficacy of electrotactile vestibular substitution in patients with peripheral and central vestibular loss. AB - Vestibular dysfunction of either central or peripheral origin can significantly affect balance, posture, and gait. We conducted a pilot study to test the effectiveness of training with the BrainPort balance device in subjects with a balance dysfunction due to peripheral or central vestibular loss. The BrainPort balance device transmits information about the patient's head position via electrotactile stimulation of the tongue. Head position data is sensed by an accelerometer and displayed on the tongue as a pattern of stimulation. This pattern of stimulation moves forward, backward, and laterally on the tongue in direct response to head movements. Users of the device were trained to use this stimulation to adjust their position in order to maintain their balance. Twenty eight subjects with peripheral or central vestibular loss were trained with the BrainPort balance device and tested using the following standardized quantitative measurements of the treatment effects: Computerized Dynamic Posturography (CDP) using the Sensory Organization Test (SOT), Dynamic Gait Index (DGI), Activities specific Balance Confidence Scale (ABC), and Dizziness Handicap Inventory (DHI). All subjects had chronic balance problems and all but one had previously participated in vestibular rehabilitation therapy. The scores on the clinical tests upon entry into the study were compared to their scores following training with the BrainPort balance device. Our results exhibit consistent positive and statistically significant improvements in balance, posture and gait. These results exceed what could normally be achieved in three to five days of traditional balance training alone. Since this was not a controlled study, we are unable to distinguish the degree to which these improvements are attributable to training with the BrainPort balance device versus the balance exercises performed by all subjects as a part of the BrainPort training sessions. Nonetheless, after training with the BrainPort balance device, all subjects demonstrated significant improvements in performance beyond what might be expected from conventional vestibular rehabilitation therapy. PMID- 18413906 TI - Improved postural control after computerized optokinetic therapy based on stochastic visual stimulation in patients with vestibular dysfunction. AB - Balance is accomplished by the congruent integration of visual, vestibular and somatosensory input and the execution of adequate control movements. With increasing age, nonlinear dynamics of central control systems become more regular. In unilateral vestibular dysfunction, sensory input to central systems is similarly less complex, because of one sided reduction of information influx. This study aimed to increase postural stability in patients with vestibular asymmetry and resulting disequilibrium by implementing a computerized visual training method relying on the principles of stochastic resonance. 24 subjects (average age 64a, 31-78a, 15 women, 9 men), with minimum 3 months of persisting disequilibrium due to vestibular dysfunction, were either treated with computerized optokinetic therapy (COKT), or solely observed. Treated patients were requested to read texts, stochastically moving in a previously defined matrix, during 10 sessions over three weeks. The Sensory Organization Test (SOT) was used for comparative posturographic measurements. COKT patients showed significant improvement in conditions 4, 6 and composite score. A significant post-therapeutic difference was seen between therapy and control groups in conditions 1, 6 and composite score. The results show a clinical benefit and we conclude COKT to be an effective rehabilitation method in patients with chronic disequilibrium. PMID- 18413907 TI - Short-term rehabilitation of patients with posttraumatic otolith disorders by auditory feedback training: a pilot study. AB - In a prospective, non-randomized pilot study, two groups of patients with dizziness for more than one year due to posttraumatic otolith disorders had been investigated. They were treated by a one-day vestibular rehabilitation training with/without auditory feedback. It could be shown that auditory feedback training for a limited period of time can reduce significantly trunk sway compared to those patients who did vestibular exercises alone without auditory feedback. This audiofeedback-based therapy seems to be a promising tool in the vestibular rehabilitation and in treating patients with otolith disorders in particular. Long-term effects cannot yet be assessed with the present data of our series. PMID- 18413908 TI - Assessment of behavioural markers of autonoetic consciousness during episodic autobiographical memory retrieval: a preliminary analysis. AB - There is ongoing theoretical debate regarding episodic memory and how it can be accurately measured, in particular if the focus should be content-based recall of episodic details or something more experiential involving the subjective capacity to mentally travel back in time and "re-live" aspects of the original event. The autonoetic subscale of the Episodic Autobiographical Memory Interview (EAMI) is presented here as a new test instrument that attempts to redress theoretical and methodological shortcomings in autobiographical memory assessment. The EAMI merges a phenomenological detail-based approach with an assessment of autonoetic consciousness, departing considerably from traditional Remember/Know paradigms used within this field. We present findings from an initial pilot study investigating the potential markers of autonoetic consciousness that may accompany episodic retrieval. Key behavioural indices of autonoetic consciousness, notably those of viewer perspective, visual imagery, and emotional re-experiencing, emerged as being inextricably bound with the level of phenomenological detail recalled and the overall re-living judgment. The autonoetic subscale of the EAMI permits conceptually refined assessment of episodic personal memories and the accompanying subjective experience of mental re-living, characteristic of episodic memory. PMID- 18413909 TI - False memories of emotional and neutral words. AB - This study used the Deese-Roediger-McDermott paradigm to investigate the direction and the extent to which emotional valence in semantic word lists influences the formation of false memories (FM). The experimental paradigm consisted of 1) a study phase (learning of neutral and negative lists of words semantically associated to a non-presented critical lure (CL), 2) a free recall phase, and 3) a recognition phase. Participants had to indicate whether the displayed item was "new" (new item or non-studied CL) or "old" (studied list item). CL associated with negative word lists elicited significantly more FM than CL associated with neutral word lists. This finding is in contrast to previous work showing that emotional words elicit fewer FM than neutral words. The results of our study also suggest that valence is capable of influencing emotional memory in terms of encoding and retrieval processes. PMID- 18413910 TI - Undifferentiation of somatic responses to emotions in a case of functional amnesia. AB - The term functional amnesia (FA) has been proposed for cases of memory impairment presenting with severe retrograde amnesia in the absence of cerebral injury or history of psychiatric disturbance. Emotional flattening has often been reported alongside FA, however the mechanism of such a modification is unknown. This study aimed to explore the emotional processing in a rare case of a patient with FA complaining of severe emotional flattening. We presented ecological dynamic video stimuli conveying strong peaceful and fearful emotions to the patient and 13 controls. We then explored their emotional responses considering both conscious emotional judgements and automatic psychophysiological responses (skin conductance) and facial muscular activity (corrugator supercilii). Both patient P.P. and controls perfectly recognized the emotions conveyed by the films. However, P.P. failed to show an increased skin conductance and corrugator activity as found in controls during fearful film extracts compared with peaceful extracts. Taken together, these finding demonstrate the presence of an emotional deficit, characterized by a failure to generate appropriate somatic responses to positive and negative stimuli. Although this altered somatic processing did not interfere with PP's explicit recognition of emotion, it modified his emotional experience, thereby constituting a possible explanation for his emotional flattening. This study therefore suggests that FA is not limited to a mnemonic impairment, but is a more complex disorder, involving also the processing of emotionally loaded experiences. PMID- 18413911 TI - Autonoetic consciousness in autobiographical memories after medial temporal lobe resection. AB - This study aims to investigate autonoetic consciousness associated with episodic autobiographical memory in patients who had undergone unilateral medial temporal lobe resection for intractable epilepsy. Autonoetic consciousness, defined as the conscious feeling of mentally travelling back in time to relive a specific event, was assessed using the Remember/Know (R/K) paradigm across different time periods as proposed in the autobiographical memory task developed by Piolino et al. (TEMPau task). Results revealed that the two patient groups (left and right temporal resection) gave reduced sense of reliving (R) responses and more familiarity (K) responses than healthy controls. This poor autonoetic consciousness was highlighted when patients were asked to justify their Remember responses by recalling sensory-perceptive, affective or spatiotemporal specific details across all life periods. These results support the bilateral MTL contribution to episodic autobiographical memory covering the entire lifespan, which is consistent with the multiple trace theory of MTL function. This study also demonstrates the bilateral involvement of MTL structures in recalling specific details of personal events characterized by autonoetic consciousness. PMID- 18413912 TI - Recognition memory and prefrontal cortex: dissociating recollection and familiarity processes using rTMS. AB - Recognition memory can be supported by both the assessment of the familiarity of an item and by the recollection of the context in which an item was encountered. The neural substrates of these memory processes are controversial. To address these issues we applied repetitive transcranial magnetic stimulation (rTMS) over the right and left dorsolateral prefrontal cortex (DLPFC) of healthy subjects performing a remember/know task. rTMS disrupted familiarity judgments when applied before encoding of stimuli over both right and left DLPFC. rTMS disrupted recollection when applied before encoding of stimuli over the right DLPFC. These findings suggest that the DLPFC plays a critical role in recognition memory based on familiarity as well as recollection. PMID- 18413913 TI - Anatomy of the episodic buffer: a voxel-based morphometry study in patients with dementia. AB - In 2000 Baddeley proposed the existence of a new component of working memory, the episodic buffer, which should contribute to the on-line maintenance of integrated memory traces. The author assumed that this component should be critical for immediate recall of a short story that exceeds the capacity of the phonological store. Accordingly, patients with Alzheimer's dementia (AD) should suffer of a deficit of the episodic buffer when immediate recall of a short story is impossible. On the other hand, the episodic buffer should be somewhat preserved in such patients when some IR can occur (Baddeley and Wilson, 2002). We adopted this logic for a voxel-based morphometry study. We compared the distribution of grey-matter density of two such groups of AD patients with and of a group of age matched controls. We found that both AD groups had a significant atrophy of the left mid-hippocampus; on the other hand, the anterior part of the hippocampus was significantly more atrophic in patients who were also impaired on the immediate prose recall task. Six out of ten patients with no immediate recall were spared at "central executive" tasks. Taken together our findings suggest that the left anterior hippocampus contributes to the episodic buffer of the revised working memory model. We also suggest that the episodic buffer is somewhat independent from the central executive component of working memory. PMID- 18413914 TI - Patterns of semantic memory impairment in Mild Cognitive Impairment. AB - Although the semantic memory impairment has been largely documented in Alzheimer's disease, little is known about semantic memory in the preclinical phase of the disease (Mild Cognitive Impairment). The purpose of this study was to document the nature of semantic breakdown using a battery of tests assessing different aspects of conceptual knowledge: knowledge about common objects, famous people and famous public events. Results indicate that all domains of semantic memory were impaired in MCI individuals but knowledge about famous people and famous events was affected to a greater extent than knowledge about objects. This pattern of results suggests that conceptual entities with distinctive and unique properties may be more prone to semantic breakdown in MCI. In summary, results of this study support the view that genuine semantic deficits are present in MCI. It could be useful to investigate the etiological outcome of patients failing or succeeding at such tests. PMID- 18413915 TI - Associative visual agnosia: a case study. AB - We report a case of massive associative visual agnosia. In the light of current theories of identification and semantic knowledge organization, a deficit involving both levels of structural description system and visual semantics must be assumed to explain the case. We suggest, in line with a previous case study, an alternative account in the framework of (non abstractive) episodic models of memory. PMID- 18413916 TI - Dopaminergic modulation of prospective memory in Parkinson's disease. AB - Growing interest is present in literature on the study of prospective memory functioning in Parkinson's disease (PD). Current data indicate that prospective memory may be impaired in PD and a relationship with general executive dysfunctioning has been suggested. However, although the dopamine dependency of cognitive dysfunction in PD has been widely investigated, poor is known on the dopaminergic modulation of PM. In the present study we explored the effect of acute administration of levodopa on the performance of a PD sample (n=20) in a time-based prospective memory task. PD patients were evaluated in the morning after a 12-hour therapy wash-out in two experimental conditions: i) after levodopa assumption ("on"); ii) without drug administration ("off"). The experimental task required to execute three uncorrelated actions after 10' for three consecutive trials. Distinct scores for the spontaneous recall of the intention to perform the actions (prospective component) and for the correct execution of the actions (retrospective component) have been computed. Results showed that in the "off" condition PD patients were selectively impaired on the prospective component of the task. However, L-dopa administration significantly improved PD patients' performance actually restoring the prospective memory deficit. These results support a critical role of dopaminergic modulation in prospective memory processes in PD patients, possibly through the replacement of dopamine levels in fronto-striatal pathways. PMID- 18413917 TI - Public events and flashbulb memories in Parkinson's disease. AB - Public events and Flashbulb memories were investigated in 12 non-demented patients with Parkinson's disease (PD) and 12 controls. Knowledge of public events and flashbulbs memories were assessed using a Famous Events Test (EVE 30). Contributions of semantic, episodic, as well as executive functioning and anterograde memory were examined. Results primarily showed that the performances of patients with PD were lower than these of controls in 4 tasks: free recall, specific questions, dating events and date recognition. They also had difficulties in finding the temporal order of 8 events. In contrast, the PD group benefited from events recognition themselves to the same extent as the controls. Secondly, the recall of flashbulb memories (FBM) was lower in the PD group than in the controls. Finally, correlations appeared in PD between the detailed recall of the events with the "recall" abilities of the MATTIS scale, possibly reflect an impairment in rebuilding memories. A positive correlation is also observed with the initiation score of the MATTIS (executive component), suggesting that the difficulties of rebuilding can be related to a dysfunction in accessing information because of a certain degree of frontal amnesia. PMID- 18413918 TI - Effect of distorted visual feedback on the sense of agency. AB - It has been hypothesized that an internal model is involved in controlling and recognizing one's own actions (action attribution). This results from a comparison process between the predicted sensory feedback of the action and its real sensory consequences. The aim of the present study is to distinguish the respective importance of two action parameters (time and direction) on such an attribution judgment. We used a device that allows introduction of discordance between the movements actually performed and the sensory feedback displayed on a computer screen. Participants were asked to judge whether they were viewing 1) their own movements, 2) their own movements modified (spatially or temporally displaced), or 3) those of another agent (i.e, the experimenter). In fact, in all conditions they were only shown their own movements either unaltered or modified by varying amounts in space or time. Movements were only attributed to another agent when there was a high spatial discordance between participants' hand movements and sensory feedback. This study is the first to show that the direction of movements is a cardinal feature in action attribution, whereas temporal properties of movements play a less important role. PMID- 18413919 TI - Age-related quantitative and qualitative changes in decision making ability. AB - The "frontal aging hypothesis" predicts that brain senescence affects predominantly the prefrontal regions. Preliminary evidence has recently been gathered in favour of an age-related change in a typically frontal process, i.e. decision making, using the Iowa Gambling Task (IGT), but overall findings have been conflicting. Following the traditional scoring method, coupled with a qualitative analysis, in the present study we compared IGT performance of 40 young (mean age: 27.9+/-4.7) and 40 old (mean age: 65.4+/-8.6) healthy adults and of 18 patients affected by frontal lobe dementia of mild severity (mean age: 65.1+/-7.4, mean MMSE score: 24.1+/-3.9). Quantitative findings support the notion that decision making ability declines with age; moreover, it approximates the impairment observed in executive dysfunction due to neurodegeneration. Results of the qualitative analysis did not reach statistical significance for the motivational and learning decision making components considered, but approached significance for the attentional component for elderly versus young normals, suggesting a possible decrease in the ability to maintain sustained attention during complex and prolonged tasks as the putative deficit underlying impaired decision making in normal aging. PMID- 18413920 TI - Studying social cognition in patients with schizophrenia and patients with frontotemporal dementia: theory of mind and the perception of sarcasm. AB - We investigated social cognition and theory of mind in patients with schizophrenia and in patients with frontotemporal dementia in order to elucidate the cognitive mechanisms involved in the breakdown of these skills in psychiatric and neurological patients. Our tasks included videotaped scenarios of social interactions depicting sincere, sarcastic and paradoxical remarks, as well as lies. We found impaired performance of the schizophrenia group on all theory of mind conditions despite their intact understanding of sincere statements. In contrast, the FTD group performed poorly only when they had to rely on paralinguistic cues indicating sarcasm or lies, and not on paradoxical remarks or sarcasm when given additional verbal cues. Our findings suggest that, while current deficits in social and interpersonal functioning in patients with FTD may reflect a decrement in previously acquired skills, similar deficits in patients with schizophrenia may reflect an altogether inadequately learned process. PMID- 18413921 TI - Prefrontal and temporo-parietal involvement in taking others' perspective: TMS evidence. AB - INTRODUCTION: Understanding the mental states of others entails a number of cognitive processes known as Theory of Mind (ToM). Behavioural and functional neuroimaging evidence suggests that prefrontal and temporo-parietal cortices are involved in these abilities. The present study was aimed at investigating the role of the dorsolateral prefrontal cortex and temporo-parietal junction in ToM by using a repetitive transcranial magnetic stimulation (rTMS) paradigm. MATERIAL AND METHODS: Eleven healthy subjects participated in the study. The experimental ToM procedure was constituted by false belief and faux-pas written stories. Subjects were evaluated in baseline condition (Sham) and after 1Hz rTMS over the left/right dorsolateral prefrontal cortex and temporo-parietal junction. A score for accuracy and response times were recorded. RESULTS: As regards false beliefs, rTMS over right prefrontal and temporo-parietal areas significantly interfered with response times (p<0.05$). The application of rTMS over right/left prefrontal and right temporo-parietal cortices also significantly worsened accuracy in the ability to take the others' perspective in faux-pas tasks as compared to Sham (p 50 kPa) was far above the favorable values, acetate was produced as well as butyrate in all pH levels tested. To determine whether some portion of the acetate is produced through another pathway such as autotrophic synthesis via H(2) dependent reduction of CO(2) or not, batch dissolved H(2) consumption rate tests were performed at pH 5.0, 5.5 and 6. The specific H(2) consumption rate was 488(+/-49) micromol/gVSS.hr at pH 6 and slightly higher than at pH 5 and 5.5. The results of continuous and batch experiments revealed that acetogenic H(2) consumption is more favorable at pH levels above 5.5 and is one of the reasons of instabilities in dark fermentative H(2) production. PMID- 18413939 TI - Impact of high saline wastewaters on anaerobic granular sludge functionalities. AB - Three UASB reactors were operated at different salinity levels in order to assess the effects on the granular sludge properties. High levels of activity inhibition were observed at sodium concentrations over 7 g Na(+)/L, which resulted in low applicable organic loading rates and VFA accumulation in reactor effluents. However, either sludge adaptation or selection for saline resistant microorganisms occurred, which could be concluded from the observed increase in the 50% activity inhibitory concentrations of sodium during continuous flow experiments. Changes in Na(+) susceptibility in time are likely to be expected when treating saline wastewaters. The latter was evidenced by the high sodium tolerance of granular methanogenic sludge coming from a full-scale industrial reactor treating such wastewater. High salinity conditions resulted in a reduced granule strength, predicting process instabilities during long term reactor operation. Batch tests showed that high sodium concentrations seemed to displace the calcium from the granular sludge, a factor known to affect anaerobic granules formation. PMID- 18413940 TI - Comparing the effluent organic matter removal of direct NF and powdered activated carbon/NF as high quality pretreatment options for artificial groundwater recharge. AB - Direct nanofiltration and nanofiltration combined with powdered activated carbon known as the PAC/NF process were tested regarding the removal of effluent organic matter for reclamation of tertiary effluent from a municipal wastewater treatment plant. They can be regarded as a promising treatment alternative for high quality water reuse applications, especially for direct injection. The total removal for DOC was above 90% with permeate concentrations below 0.5 mg/l. Size exclusion chromatography and fluorescence EEM proved to trace origin of the organic matter even in low concentration ranges. The type and dosage of adsorbent influences the process performance significantly and allows process optimization. PMID- 18413941 TI - Dimensioning of membrane bioreactors for municipal wastewater treatment. AB - A basic procedure is proposed for dimensioning membrane bioreactors that is generally applicable. It evaluates the required membrane surface with particular consideration of loading combinations and hydraulic loading characteristics; it also takes into account ranges of minimum temperatures and corresponding fluxes as given by suppliers. The procedures, while likely to require further improvement, should make MBR design more transparent and aid the comparison of design variants. PMID- 18413942 TI - Characterization of anaerobic granular sludge developed in UASB reactors that treat ethanol, carbohydrates and hydrolyzed protein based wastewaters. AB - In this work, granules developed from UASB reactors that treat different types of wastewaters (ethanol, carbohydrates and protein-based synthetic wastewaters) were studied. Granule parameters (size distribution; density; settlement characteristics; elemental composition; acidogenic and methanogenic activities) were analyzed along with micro-organisms identified by FISH to better understand granule behavior and its formation process. Micro-organisms distributions in anaerobic granules are highly dependent on the type of treated wastewater. Granules developed in a UASB reactor that treats wastewater with a high content of carbohydrates presented high acidogenic bacteria colonization. Members of Methanosaetaceae were the dominant methanogens in the studied granules, and Methanobacteriales appear to be co-dominant in the granules developed with carbohydrates and protein-based wastewaters. PMID- 18413943 TI - Microbial challenge-testing of treatment processes for quantifying stormwater recycling risks and management. AB - Pathogenic microorganisms have been identified as the main human health risks associated with the reuse of treated urban stormwater (runoff from paved and unpaved urban areas). As part of the Smart Water initiative (Victorian Government, Australia), a collaborative evaluation of three existing integrated stormwater recycling systems, and the risks involved in non-potable reuse of treated urban stormwater is being undertaken. Three stormwater recycling systems were selected at urban locations to provide a range of barriers including biofiltration, storage tanks, UV disinfection, a constructed wetland, and retention ponds. Recycled water from each of the systems is used for open space irrigation. In order to adequately undertake exposure assessments, it was necessary to quantify the efficacy of key barriers in each exposure pathway. Given that none of the selected treatment systems had previously been evaluated for their treatment efficiency, experimental work was carried out comprising dry and wet weather monitoring of each system (for a period of 12 months), as well as challenging the barriers with model microbes (for viruses, bacteria and parasitic protozoa) to provide input data for use in Quantitative Microbial Risk Assessment. PMID- 18413944 TI - Fate and transport of faecal contamination microbial indicators, pathogenic protozoa and Campylobacter in the artificially recharged fractured aquifer of Salento, Italy. AB - The study investigates the fate and transport of microorganisms introduced by artificial groundwater recharge at the Nardo fractured aquifer in Salento, Italy. Microbial indicators of faecal contamination, parasitic protozoa (Giardia and Cryptosporidium) and pathogenic bacteria (Campylobacter spp.), were monitored into injected water and groundwater to test the efficiency of the "natural disinfection" into the fractured aquifer. A remarkable decrease of microbial indicators and pathogens was observed suggesting that pathogens removal or inactivation may be possible during water flow in fractured aquifer. The recently described PNA probe CJE195 (Lehtola et al. 2005) was utilised for the rapid and specific detection of Campylobacter spp. by fluorescence in situ hybridization (FISH) after enrichment. FISH results were consistent with those of traditional cultural method (ISO 17995) applied in parallel: time required for Campylobacter identification was reduced of 4 days. PMID- 18413945 TI - Analysis of the methodology to determine anaerobic toxicity: evaluation of main compounds present in wastewater treatment plants (WWTPs). AB - The influence of the concentration of biomass on the level of inhibition and anaerobic degradation kinetics in batch systems was studied with toxic compounds that can generate destabilization in the operation of sludge anaerobic digesters. The compounds were grouped in four families; long chain fatty acids, polycyclic aromatic hydrocarbons, linear alkylbenzene sulphonates and organic acids. For the organic acids, there is no effect due to the biomass concentration variation, therefore it is a competitive inhibition; but that doesn't happen with the remaining compounds, where there is a dependence on the complexity of their structure, becoming a non-competitive inhibition. In addition, it was observed that the degradation kinetics is affected, whether diminishing the methane production (polycyclic aromatic hydrocarbons, linear alkylbenzene sulphonates, organics acids) or increasing the initial latency time (long chain fatty acids) without this becoming an obstacle to obtain the maximum methane productions for the latter ones. PMID- 18413946 TI - Treatment of textile dyes in two-phase and single-phase anaerobic bio-treatment systems. AB - This research integrates two different concepts of anaerobic biotechnology- two phase anaerobic treatment and anaerobic granular sludge bed technology, in treatment of colored wastewaters from textile industries. Four anaerobic reactors based on upflow anaerobic sludge blanket (UASB) technology were used as acid reactors and an expanded granular sludge bed (EGSB) reactor was used as a methane reactor. A conventional single-phase anaerobic reactor, working on EGSB technology was run in parallel to compare the performances of the two systems. Reactors were operated at different hydraulic retention times. The results from the study, which span over a period of 400 days, indicated that the two-phase system produces a higher quality of effluent in terms of color, COD and suspended solids than single-phase anaerobic treatment when operated under similar conditions. Alkalinity requirement of two-phase system was also observed to be lower than that of single-phase system which is important regarding design consideration. PMID- 18413947 TI - Influence of effluent-recirculation condition on the process performance of expanded granular sludge bed reactor for treating low strength wastewater. AB - A 2.0 L volume of EGSB reactor was operated at 20 degrees C for more than 500 days with 0.3-0.4 g COD/L of sucrose base wastewater to investigate the influence of effluent-recirculation on the process performance. At the start up period, the reactor was operated in EGSB mode with 5 m/h upflow velocity by continuous effluent recirculation. The COD loading was set to 7.2-9.6 kg COD/m(3) day with HRT of 1 hour. However, in this mode, EGSB reactor exhibited insufficient COD removal efficiency, i.e., 50-60%. Therefore, UASB mode (without recirculation, 0.7 m/h upflow velocity) was used for 30 minutes in every 40 minutes cycle to increase the COD concentration in the sludge bed. As a result, an excellent process performance was shown. The COD removal efficiency increased from 65% to 91% and the reactor could maintain a good physical property of retained sludge (sludge concentration: 33.4 g VSS/L and SVI: 25 mL/g VSS). Furthermore, retained sludge possessed sufficient level of methanogenic activity at 20 degrees C. PMID- 18413948 TI - Treatment of petroleum production wastewater for reuse in secondary oil recovery. AB - Petroleum production wastewater is highly saline and contains large amounts of Ca, Mg, sulphides and hydrocarbons. The reuse of this wastewater in the secondary oil recovery can provide pollution prevention and water conservation benefits. Injection of water to the oil deposits is a frequently used method for secondary oil recovery. This operation is performed at high pressures and temperatures, because of which a suitable water quality is required to avoid deposit formation, scaling and clogging effects. The objective of this study was to select the best treatment system for the oil production wastewater, generated in one of the Mexican oil extraction facilities, for the implementation of this kind of reuse by injecting the treated water to sand formations with 12-15% porosity. A complete characterization of the petroleum production wastewater was done. Based on laboratory tests, three basic treatment options were suggested and evaluated in a pilot plant. The most suitable treatment was determined by injecting the different treated waters in samples of the real formations. The selected system consists in softening, followed by oxidation, decarbonation and filtration. This train allowed 99.8% hardness removal, a complete S(2-) removal, as well as 99% TSS, 78% TOC, 98% Sr, 86% Ba, 51% Si and 17% Fe removals. PMID- 18413949 TI - Reuse of industrial wastewater for the irrigation of ornamental plants. AB - This paper describes the results of experimental activities carried out for verifying the possibility of reusing reclaimed wastewater originated from textile (70%) and domestic (30%) activities for the irrigation of container-grown ornamental shrubs. Aspects that concern the refinery treatment of reclaimed wastewater and the effect of irrigation on some ornamental plant species were investigated. An experimental site consisting of a refinery treatment pilot plant (filtration and disinfection) and an agronomic experimental area was set-up. The combined treatment of PAA and UV, used for the disinfection, showed to be very effective for inactivation of E. coli with most of PAA and UV dose combinations able to assure total inactivation. The plants (Buxus, Photinia, Pistacia and Viburnum), sprinkle and drip irrigated with well water (WW), reclaimed wastewater (RW) and a water mixed (MW) between reclaimed wastewater and well water (1:1 by vol), showed interesting results. A similar growth among different treatments was achieved for Buxus and Pistacia, while Viburnum and Photinia plants showed a higher sensibility to MW and RW. Photinia, in particular, turned out to be very sensitive to sprinkle irrigation with the reclaimed water, while the drip irrigation had no such bad effects, as reported in previous works. PMID- 18413950 TI - Urban versus regional--how public attitudes to recycled water differ in these contexts. AB - This paper reports findings from a comparative study which investigated public attitudes to recycled water in two Australian locations both in the state of Victoria: the capital city, Melbourne, and Bendigo, an urban regional centre. Two commercial buildings were used as case studies, one at each location. These buildings will soon be using recycled water for non-potable uses. The study was facilitated by an on-line survey of future occupants of both buildings to gauge their attitudes to recycled water use. Specifically the paper reports on happiness/willingness to use recycled water for various uses and attitudinal factors which were found to influence this. The circumstances for potable water availability and recycled water use differ in Melbourne and Bendigo, making this study a significant contribution to understanding public acceptance of recycled water use in these different contexts. No significant difference in happiness to use recycled water was found between locations. However, prior experience (use) of recycled water was found to be a significant and positive factor in facilitating happiness/willingness to use recycled water, particularly for closer to personal contact uses such as showering and drinking. Various attitudinal and demographic variables were found to influence happiness to use recycled water. Results indicate it is not just the locational context of water availability that influences happiness to use recycled water, but a person's experience and particular perceptions that will facilitate greater willingness to use recycled water. PMID- 18413951 TI - Evaluation of state variable interface between the Activated Sludge Models and Anaerobic Digestion Model no 1. AB - For plant wide modelling of wastewater treatment, it is necessary to develop a suitable state variables interface for integrating state of the art models of ASM and ADM1. ADM1 currently describes such an interface, however, its suitability needs to be experimentally evaluated. In this study, we characterised activated sludge under aerobic and anaerobic conditions to obtain representative state variables for both models. ASM state variables of X(S), X(H) and X(I) (as obtained from aerobic tests) and ADM1 state variables of X(C) and X(I) (as obtained from anaerobic tests) were then correlated to assess the suitability of current interface. Based on the seven datasets of this study and seven datasets from literatures, it was found that in general ASM state variables were well correlated to the state variables of ADM1. The ADM1 state variable of X(C) could be correlated to the sum of state variables of X(S) and X(H), while X(I) in both the models showed direct correspondence. It was also observed that the degradation kinetics of X(C) under anaerobic condition could be better described by individual degradation kinetics of X(S) and X(H). Therefore, to establish a one to one correspondence between ASM and ADM1 state variables and better description of degradation kinetics in ADM1, replacing the composite variable of X(C) by the state variables of X(S) and X(H) is recommended. PMID- 18413952 TI - An integrated wastewater reuse concept combining natural reclamation techniques, membrane filtration and metal oxide adsorption. AB - In a Sino-German research project, a sustainable water reclamation concept was developed for different applications of municipal water reuse at the Olympic Green 2008 in Beijing, China. Results from pilot-scale experiments in Beijing and Berlin show that selective nutrient removal by adsorption onto granular ferric hydroxide (GFH) after a membrane bioreactor (MBR) can maintain a total phosphorus concentration of <0.03 microg L(-1) P, thus preventing eutrophication of artificial lakes. Operation time of GFH adsorption columns can be extended by regeneration using sodium hydroxide solution. A subsequent ultrafiltration (UF) membrane after bank filtration creates an additional barrier for pathogens and allows for further urban reuse applications such as toilet flushing. Short term bank / bio-filtration prior to UF is shown to effectively remove biopolymers and reduce membrane fouling. PMID- 18413953 TI - Inhibition of sulfide generation by dosing formaldehyde and its derivatives in sewage under anaerobic conditions. AB - Hydrogen sulfide emission in sewers is associated with toxicity, corrosion, odor nuisance and a lot of costs. The possibility to inhibit sulfide generation by formaldehyde and its derivatives (paraformaldehyde and urea formaldehyde) has been evaluated under anaerobic conditions. The impact of formaldehyde on an activated sludge system and an appraisal of the economic aspects are also presented. The optimum dosage to inhibit sulfide generation in sewage was 12-19 mg L(-1) formaldehyde. The dosages of 32 mg L(-1) paraformaldehyde or 100 mg L( 1) urea formaldehyde were not capable of inhibiting sulfide generation in sewage. The impact of 19 mg L(-1) formaldehyde on activated sludge system was negligible in terms of COD removal, nitrification rate and oxygen uptake rate. PMID- 18413954 TI - Use of poly-beta-hydroxy-butyrate as a slow-release electron donor for the microbial reductive dechlorination of TCE. AB - In situ anaerobic reductive dechlorination, using slow-release electron donors, is emerging as an effective and sustainable (low-cost and low-maintenance) technology to remediate aquifers contaminated by chloroethenes. In the present study, we investigated the use of poly-beta-hydroxy-butyrate (PHB), a fully biodegradable polymer, as a slow-release source of hydrogen and acetate for the reductive dechlorination of trichloroethene (TCE). Results of this study indicated that TCE dechlorination in PHB-amended microcosms was 2.3-times higher than in non-amended controls. This higher activity was explained by a higher H(2) level in PHB-amended microcosms. As usual, acetate was the major sink (approximately 90%) of reducing equivalents available from PHB degradation, whereas no acetotrophic dechlorination was observed. PMID- 18413955 TI - Use of nanofiltration for potable water from an aquifer recharged with wastewater. AB - This paper presents the comparison of nine nanofiltration membranes to treat water coming from an aquifer recharged with wastewater and used as municipal supply in the Tula Valley, Mexico. The comparison was made based on (a) the amount of water produced; (c) the capability to produce a <1 mg TOC/L effluent without entirely eliminating salts, (b) the removal of specific organic and microbiological pollutants, and (c) the reduction of toxicity and mutagenicity from water. From the tested membranes, only four produce an effluent with <1 TOC mg/L, and three totally retained dibutyl phthalate, diethyl phthalate and hydroxytoluene butylate. Influent mutagenicity (Ames test) was negative but these was a certain degree of toxicity when Tetrahymena pyriformis was used as indicator. Toxicity was partially reduced by some of the NF membranes. The best membrane had a flux of 95 L m(-2)h(-1) and removal efficiencies of 98% for TOC, 92% for AUV(254), and 92% for TDS. The permeate had a final hardness of 76 mg/L and an alkalinity of 124 mg/L. Additionally, this membrane removed totally specific organic compounds, total and fecal coliforms and almost all the somatic coliphages. PMID- 18413956 TI - The combined performance of UV light and chlorine during reclaimed water disinfection. AB - The combined effects of disinfectant agents on the microbiological quality of reclaimed water produced by two full-scale water reclamation plants in Catalonia, Spain, were examined in this work. All the disinfectant treatments tested led to the absence, or near absence, of E. coli in 100 mL samples of water, with log reductions of more than 3 log u. Hypochlorite reduced the bacterial concentrations. However, ultraviolet light was more effective than hypochlorite at reducing the concentrations of bacteriophages, viruses and pathogenic protozoa such as Cryptosporidium spp. We conclude that a combination of these two disinfectant agents is effective in protecting public health, as each agent acts to a different degree against the different groups of microorganisms studied. Further studies should investigate the combined action of disinfectant agents at water reclamation plants with ultraviolet light equipment in more favourable working conditions in order to assess their capacity to inactivate microorganisms. PMID- 18413957 TI - Framework for assessment of performance of soil aquifer treatment systems. AB - Removal of organic matter is a critical parameter in soil aquifer treatment (SAT) as it governs and influences the removal of other contaminants by biodegradation namely trace organics, nitrogen species and microbes. A framework for analysis and prediction of the performance of SAT systems with respect to removal of organic matter under different water quality and process conditions was developed based on an extensive literature review and data analysis. Guidelines were developed to make preliminary estimates of the removal of organic matter during SAT using primary, secondary and tertiary effluents from wastewater treatment plants. These guidelines can be used as a quick tool to analyze the performance of existing SAT systems and serve as a decision support tool for feasibility studies and to save time for further detailed experimentation and design of SAT systems. Furthermore, effects of soil type and redox conditions on organic matter removal during SAT were also analyzed. PMID- 18413958 TI - Bioassays and biomarkers for ecotoxicological assessment of reclaimed municipal wastewater. AB - The aim of this work was to examine the ecotoxicity of reclaimed wastewater by the use of bioassays and the determination of immunological parameters. Secondary and tertiary mucicipal wastewater samples were examined for their physicochemical and microbiological characteristics as well as for their endotoxin concentrations. The ecotoxicological characteristics were assessed by a battery of bioassays, using Vibrio fischeri, Daphnia magna and Tetrahymena thermophilla as test species and phytotoxicity. The mitogenic responses of mouse splenocytes were as well used as bioassay. The cytokines of IL-1, IL-2, IL-10, IFNgamma and TNFalpha, were also determined in the supernatant of splenocyte cultures and served as molecular biomarkers. All bioassays exhibited decrease of the ecotoxicological responses after tertiary treatment. However, mitogenic responses were proved to be more sensitive. IL-1 increased, while IL-2 production was unaffected. The fact that IL-10 production increased in response to secondary treated effluents in conjunction with the increased endotoxin levels, suggest Th2 type immune responses. Although results obtained from the toxicity bioassays after the tertiary treatment showed comparable results to those of controls, cytokine levels indicated the induction of immune response even after tertiary treatment. Consequently, cytokine production could be used as a sensitive biomarker for the evaluation of treatment efficiency of the reclaimed wastewaters intended for reuse. PMID- 18413959 TI - The use of ozone during advanced primary treatment of wastewater for its reuse in agriculture: an approach to enhance coagulation, disinfection and crop productivities. AB - The purpose of this work was to evaluate the effect that ozone (O(3)) has on the advanced primary treatment (APT) and disinfection of wastewaters for their reuse in agriculture. The enhancement and optimization of APT was done by applying low O(3) doses during coagulation. By applying an ozone dose of 2 mg/L during APT, the required coagulant dose may be reduced by up to 25% to achieve a similar turbidity removal (and up to 50% for total suspended solids removal), when compared to a conventional APT treatment. When the same coagulant dose was applied (60 mg/L), the volume of settleable solids was reduced from 31.0 to 25.5 mL/L, and the settling velocity increased from 0.111 to 0.139 m/min. Disinfection was also improved by the use of ozone, which leads to better plant germination rates than when using chlorination, because of reduced toxicity of the ozonated effluents. Additionally, helminth eggs content was reduced by applying ozone. PMID- 18413961 TI - Work organization and workers' safety and health. PMID- 18413962 TI - Effects of VDT workstation lighting conditions on operator visual workload. AB - Industrial lighting covers a wide range of different characteristics of working interiors and work tasks. This study investigated the effects of illumination on visual workload in visual display terminal (VDT) workstation. Ten college students (5 males and 5 females) were recruited as participants to perform VDT signal detection tasks. A randomized block design was utilized with four light colors (red, blue, green and white), two ambient illumination levels (20 lux and 340 lux), with the subject as the block. The dependent variables were the change of critical fusion frequency (CFF), visual acuity, reaction time of targets detection, error rates, and rating scores in a subjective questionnaire. The study results showed that both visual acuity and the subjective visual fatigue were significantly affected by the color of light. The illumination had significant effect on CFF threshold change and reaction time. Subjects prefer to perform VDT task under blue and white lights than green and red. Based on these findings, the study discusses and suggests ways of color lighting and ambient illumination to promote operators' visual performance and prevent visual fatigue effectively. PMID- 18413963 TI - Epidemiological study of low back pain and occupational risk factors among taxi drivers. AB - A survey of taxi drivers was conducted to determine the actual situation of drivers' low back pain (LBP). The survey was carried out in October 2002, the target drivers were asked to complete a questionnaire which contains questions regarding physique of drivers, demographic features, working conditions, office environment, health conditions, the presence of low back pain, the level of low back pain based on Visual Analogue Scale and Roland-Morris Disability Questionnaire score. As a result, the total number of valid responses was 1,334 and the response rate was 71 percent, and the 1-wk prevalence of LBP was 20.5 percent of respondents. Regarding 275 subjects with LBP, Visual Analogue Scale (VAS) averaged 4.3. There was a positive weak correlation between VAS and Roland Morris Disability Questionnaire score (R=0.41). And Logistic regression analysis was performed to examine the relationship between LBP and occupational factors, the results suggested following items as risk factors; such as history of LBP, suffering from fatigue, diseases other than LBP and smoking habit. PMID- 18413964 TI - Subjective scaling of hand-arm vibration. AB - The purpose of this research was to establish a scale for comfort with regard to hand-arm vibration using the category judgment method and to validate the frequency-weighting method of the ISO 5349-1 standard. Experiments were conducted using random signals as stimuli. These stimuli consisted of three types of signal, namely designated stimulus F, with flat power spectrum density (PSD) ranging from 1 to 1,000 Hz, stimulus H with PSD which became 20 dB higher at 1,000 Hz than at 1 Hz, and stimulus L that had a PSD 20 dB lower at 1,000 Hz. These stimuli were selected from the specific spectrum patterns of hand-held vibration tools. These signals were modified by the Wh frequency weighting in accordance with ISO 5349-1, and the R.M.S. values were adjusted to be equal. In addition, the signal levels were varied over a range of five steps to create 15 kinds of individual stimuli. The subjects sat in front of a vibrator and grasped the mounted handle which exposed them to vertical vibrations after which they were asked to choose a numerical category to best indicate their perceived level of comfort (or otherwise) during each stimulus. From the experimental results of the category judgment method, the relationship between the psychological values and the frequency-weighted R.M.S. acceleration according to the ISO 5349-1 standard was obtained. It was found that the subjective response scaling of hand arm vibration can be used for design-objective values of hand-held tool vibration. PMID- 18413965 TI - Modeling of a seated human body exposed to vertical vibrations in various automotive postures. AB - Although much research has been devoted to constructing specific models or to measuring the response characteristics of seated subjects, investigations on a mathematical human model on a seat with a backrest to evaluate vehicular riding comfort have not yet attracted the same level of attention. For the responses of a seated body to vertical vibrations, mathematical models of the mechanisms must be at least two-dimensional in the sagittal plane. In describing the motions of a seated body, two multibody models representative of the automotive postures found in the literature were investigated, one with and the other without a backrest support. Both models were modified to suitably represent the different automotive postures with and without backrest supports, and validated by various experimental data from the published literature pertaining to the same postural conditions. On the basis of the analytical study and the experimental validation, the fourteen-degrees-of-freedom model proposed in this research was found to be best fitted to the test results; therefore, this model is recommended for studying the biodynamic responses of a seated human body exposed to vertical vibrations in various automotive postures. PMID- 18413967 TI - The influence of particle size and composition on the quantification of airborne quartz analysis on filter paper. AB - Fine particle quartz is considered as a lung carcinogen and therefore its monitoring in the workplace is currently required by government legislation across the world. In order to enforce the health and safety standards, techniques that can accurately determine the amounts airborne quartz are necessary. Currently, two techniques dominate quartz analysis--the infrared (IR) spectrophotometric analysis and the X-ray diffraction (XRD) method. Unfortunately, there are limitations associated with both techniques in terms of their analysis accuracy and this article discusses both these methods. The study showed that typical quarry quartz samples have a broad particle size distribution where up to 20% of the finer particles in the bulk sample material contain significant amounts of muscovite, kaolinite, rutile and others. This implies that quarry silica particles with a mean diameter of less than 5 microm contain significant amounts of other materials that are not quartz, and hence, would significantly influence the quantification of the particle composition analysis on filter papers used for air monitoring work places. The results of the study suggest that analysis of the fine fraction of a manufactured sample in a particular work environment should be considered as a reference standard for the quantification of the amount of airborne fine particulates an employee could be exposed to. The study also showed that the IR method is more sensitive and less subjective to sample preparation than the XRD method of analysis. PMID- 18413966 TI - Effect of manual feeding on the level of farmer's exposure to airborne contaminants in the confinement nursery pig house. AB - The objective of the study is to demonstrate an effect of manual feeding on the level of farmer's exposure to airborne contaminants in the confinement nursery pig house. The levels of all the airborne contaminants besides respirable dust, total airborne fungi and ammonia were significantly higher in the treated nursery pig house with feeding than the control nursery pig house without feeding. Although there is no significant difference in respirable dust and total airborne fungi between the treatment and the control, their concentrations in the treated nursery pig house were also higher than the control nursery pig house. The result that the level of ammonia in the treated nursery pig house is lower than the control nursery pig house would be reasoned by the mechanism of ammonia generation in the pig house and adsorption property of ammonia to dust particles. In conclusion, manual feeding by farmer increased the exposure level of airborne contaminants compared to no feeding activity. PMID- 18413968 TI - Factors of occupational injury: a survey in a chemical company. AB - Chemical industries being the seat of dangerous occurrences frequently resulting in injuries, an occupational injury surveillance study was initiated involving 307 permanent and 419 temporary workers in a chemical company to understand the contribution of different possible factors on injury causation. Risk calculation was undertaken in relation to every individual factor using univariate and multivariate analysis techniques. Workers of lower age were found to be more susceptible to accidents (as evidenced by negative correlation coefficient), though non-significantly. Lower job duration (experience) had a significant impact on injury causation (correlation coefficient -0.5115, p<0.05). Alcohol habit could not show any significant impact but smoking/chewing habit showed significant effect (OR, 7.29: 95% CI, 3.88-9.33) on accident occurrence. Nature of job had no significant impact but nature of employment was found to have considerable effect on the causation of injuries. Temporary nature of employment was at greater risk (OR, 2.51: 95% CI, 1.42-3.77) in comparison to permanent workers. PMID- 18413969 TI - A pilot study testing the dimensions of safety climate among Japanese nurses. AB - To investigate the dimensions of safety climate among Japanese nurses, an anonymous self-administered questionnaire survey was conducted. The subjects involved in the survey included 293 full-time nurses (registered nurses and licensed practical nurses) working in a public hospital, excluding directors of nursing. A total of 221 of the 293 nurses answered the questionnaires. Among 221 questionnaires, the questionnaires, which had missing values in the question items used in this study, were excluded from the analyses. Consequently, a total of 201 questionnaires were analyzed. The average age of the subjects was 34.7 yr. As a result of exploratory factor analysis, 5 factors were extracted as follows: intellectual development regarding medical safety among nurses, accumulated fatigue, nursing conditions, supervisors' attitudes, and communication with physicians. All the values of Cronback's coefficient alpha among these 5 factors were between 0.804 and 0.892. As a result of the confirmatory factor analysis of the 5 factors, the value of the GFI (Goodness of Fit Index) was 0.868. The value of the CFI (Comparative Fit Index) was 0.943. The value of the RMSEA (Root Mean Square Error of Approximation) was 0.062. The results of this study will contribute to the investigation of the dimensions of a nurses' safety climate scale in the future. The associations between the dimensions of the safety climate and the motivation to work toward improving patients' safety among Japanese nurses will need to be examined, as will those between the dimensions of the safety climate and actual clinical mistakes. PMID- 18413970 TI - Chemical exposures in research laboratories in a university. AB - Research laboratories in a university were investigated for air-borne levels of legally designated organic solvents and specified chemical substances. Repeated surveys in 2004-5 (four times in the two years) of about 720 laboratories (thus 2,874 laboratories in total) revealed that the solvent concentrations were in excess of the Administrative Control Levels only in a few laboratories (the conditions improved shortly after the identification) and none with regard to specified chemicals. Thus, working environments were in Control Class 1 in almost all (99.5%) laboratories examined. Such conditions were achieved primarily by extensive installation and use of local exhaust systems. The survey further revealed that types of chemicals used in research laboratories were extremely various (only poorly covered by the regulation) whereas the amounts of each chemical to be consumed were quite limited. For protection of health of researchers (including post- and under-graduate students) in laboratories, therefore, it appeared more appropriate to make personal exposure assessment rather than evaluation of levels of chemicals in air of research laboratories. Considering unique characteristics of research activity, it is important to educate each researcher to make his/her own efforts to protect his/her health, through supply of knowledge on toxicity of chemicals as well as that on proper use of protective equipments including exhaust chambers. PMID- 18413971 TI - Oxidative damage of workers in secondary metal recovery plants affected by smoking status and joining the smelting work. AB - In Taiwan, secondary copper smelters and zinc recovery plants primarily utilize recovering metal from scrap and dross, and handles mostly fly ash and slag with high temperature to produce ZnO from the iron and steel industry. The materials may contain organic impurities, such as plastic and organic chloride chemicals, and amounts of polychlorinated dibenzo-p-dioxins and polychlorinated dibenzofurans (PCDD/Fs) are produced during the smelting process. Therefore, secondary metal recovery industries are major emission sources of PCDD/Fs, which may have been demonstrated to elicit oxidative stress and to involve the production of plasma malondialdehyde (MDA). Many studies have also indicated that the intake of antioxidants, smoking, age and exposure to environmental pollutants may be implicated to DNA damage or lipid peroxidation. This study therefore aims to elucidate the roles of occupational exposure like joining the smelting work, age, smoking and alcohol status, and antioxidant intake on oxidative damage in secondary metal recovery workers in Taiwan. 73 workers were recruited from 2 secondary metal recovery plants. The analysis of 8-hydroxydeoxyguanosine (8-OH dG) in urine, DNA strand breakage (comet assay) and lipid peroxidation (MDA) in blood samples were completed for all of the workers. The results showed that the older subjects exhibited significantly lower levels of 8-OH-dG and MDA than younger subjects. Our investigation also showed that working departments were in related to plasma MDA and DNA strand breakage levels of nonsmokers, however, the observation become negligible in smokers. And it is implicated that cigarette type might affect 8-OH-dG levels in secondary metal recovery workers. Since, adding to results above, the MDA level in production workers was significantly higher than those in managerial departments, it is important for the employers to make efforts on improving occupational environments or serving protective equipments to protect workers in secondary metal recovery factories. PMID- 18413972 TI - Changes of subjective stress and stress-related symptoms after a merger announcement: a longitudinal study in a merger-planning company in Japan. AB - To investigate the influences of a merger on employees in a Japanese company, changes of subjective stress and stress-related symptoms after a merger announcement in a major Japanese financial company were explored using longitudinal study surveys. Seventy-one participants responded to the first and second questionnaires, consisting of stress and symptoms, personal characteristics, lifestyle, medical examination, and work-related factors. After the merger announcement, the prevalence of subjective stress, anxiety, and impatience increased significantly from 46.5% to 78.9%, 18.3% to 40.8%, and 15.9% to 29.0%, respectively. The study suggests that subjective stress and stress related symptoms may increase after a merger announcement. To reduce the negative impact of mergers, employers are expected to provide mental health-promotion programs in a merger-planning company. PMID- 18413973 TI - A pilot study of gene expression analysis in workers with hand-arm vibration syndrome. AB - The purpose of this pilot study was to examine differences in gene expressions by cDNA microarray analysis of hand-arm vibration syndrome (HAVS) patients. Vein blood samples were collected and total RNA was extracted. All blood samples were obtained in the morning in one visit after a standard light breakfast. We performed microarray analysis with the labeled cDNA prepared by reverse transcription from RNA samples, using the Human CHIP version 1 (DNA Chip Research Inc, Yokohama, Japan). There are 2,976 genes on the chip, and these genes were selected from a cDNA library prepared with human peripheral white blood cells (WBC). Different gene levels between the HAVS patients and controls, and between groups of HAVS with different levels of symptoms, were indicated by the randomized variance model. The most up-regulated genes were analyzed for their possible functions and association with the occurrence of HAVS. From the results of this pilot study, although the results were obtained a limited number of subjects, it would appear that cDNA microarray analysis of HAVS patients has potential as a new objective method of HAVS diagnosis. Further research is needed to examine the gene expression with increased numbers of patients at different stages of HAVS. PMID- 18413974 TI - Effectiveness of a head wash cooling protocol using non-refrigerated water in reducing heat stress. AB - Envisioning a cooling method and aiming at maximum feasibility and simplicity, we designed an experimental intervention-control study based on non-refrigerated water usage, consisting of pouring 2 l of 23.0 degrees C water simultaneously on head and hands for one minute, after every 20 min of exertion. The subjects were 11 fit male individuals between 19 and 26 yr old. Each individual participated in one control and one intervention measurement in a climatic chamber at 35 degrees C and 60% humidity (31.5 degrees C WBGT) on different days. Heart rate, rectal, esophageal, skin and external ear canal temperatures were monitored constantly. Each experiment consisted of 10 min of basal recording followed by 3 intervals of 20 min of cycling and 15 min of rest. Stabilometry and visual reaction time tests were performed before and after each resting period. A questionnaire evaluating equilibrium, concentration, alertness and tiredness was administered at the beginning and at the end of every experiment. Paired t-test analysis revealed significant improvements in subjective parameters (all p<0.05), as well as skin (p<0.05), external ear canal (p<0.01) and esophageal (p<0.05) temperatures during the rest periods. Repeated measurement analysis of variance revealed significant cooling in all the aforementioned temperatures except the esophageal temperature (p=0.28). Other parameters were not significantly different. Our findings indicate that this method has subjective and physiologic positive effects, and thus can be used as a complementary low cost method to cool subjects safely. PMID- 18413975 TI - The association between self-reported periodontitis and coronary heart disease- from MY Health Up Study--. PMID- 18413976 TI - Effects of locally targeted heavy-ion and laser microbeam on root hydrotropism in Arabidopsis thaliana. AB - Classical studies on root hydrotropism have hypothesized the importance of columella cells as well as the de novo gene expression, such as auxin-inducible gene, at the elongation zone in hydrotropism; however, there has been no confirmation that columella cells or auxin-mediated signaling in the elongation zone are necessary for hydrotropism. We examined the role of root cap and elongation zone cells in root hydrotropism using heavy-ion and laser microbeam. Heavy-ion microbeam irradiation of the elongation zone, but not that of the columella cells, significantly and temporarily suppressed the development of hydrotropic curvature. However, laser ablation confirmed that columella cells are indispensable for hydrotropism. Systemic heavy-ion broad-beam irradiation suppressed de novo expression of INDOLE ACETIC ACID 5 gene, but not MIZU-KUSSEI1 gene. Our results indicate that both the root cap and elongation zone have indispensable and functionally distinct roles in root hydrotropism, and that de novo gene expression might be required for hydrotropism in the elongation zone, but not in columella cells. PMID- 18413977 TI - Formation of clustered DNA damage after high-LET irradiation: a review. AB - Radiation can cause as well as cure cancer. The risk of developing radiation induced cancer has traditionally been estimated from cancer incidence among survivors of the atomic bombs in Hiroshima and Nagasaki.(1)) These data provide the best estimate of human cancer risk over the dose range for low linear energy transfer (LET) radiations, such as X- or gamma-rays. The situation of estimating the real biological effects becomes even more difficult in the case of high LET particles encountered in space or as the result of domestic exposure to alpha particles from radon gas emitters or other radioactive emitters like uranium-238. Complex DNA damage, i.e., the signature of high-LET radiations comprises of closely spaced DNA lesions forming a cluster of DNA damage. The two basic groups of complex DNA damage are double strand breaks (DSBs) and non-DSB oxidative clustered DNA lesions (OCDL). Theoretical analysis and experimental evidence suggest an increased complexity and severity of complex DNA damage with increasing LET (linear energy transfer) and a high mutagenic or carcinogenic potential. Data available on the formation of clustered DNA damage (DSBs and OCDL) by high-LET radiations are often controversial suggesting a variable response to dose and type of radiation. The chemical nature and cellular repair mechanisms of complex DNA damage have been much less characterized than those of isolated DNA lesions like an oxidized base or a single strand break especially in the case of high-LET radiation. This review will focus on the induction of clustered DNA damage by high-LET radiations presenting the earlier and recent relative data. PMID- 18413978 TI - Suppressing effect of low-dose gamma-ray irradiation on collagen-induced arthritis. AB - We previously reported attenuation of autoimmune disease by low-dose gamma-ray irradiation in MRL-lpr/lpr mice. Here, we studied the effect of low-dose gamma ray irradiation on collagen-induced arthritis (CIA) in DBA/1J mice. Mice were immunized with type II collagen, and exposed to low-dose gamma-rays (0.5 Gy per week for 5 weeks). Paw swelling, redness, and bone degradation were suppressed by irradiation, which also delayed the onset of pathological change and reduced the severity of the arthritis. Production of tumor necrosis factor-alpha, interferon gamma, and interleukin-6, which play important roles in the onset of CIA, was suppressed by the irradiation. The level of anti-type II collagen antibody, which is essential for the onset of CIA, was also lower in irradiated CIA mice. The population of plasma cells was increased in CIA mice, but irradiation blocked this increase. Since regulatory T cells are known to be involved in suppression of autoimmune disease, the population of CD4(+)CD25(+)Foxp3(+) regulatory T cells was measured. Intriguingly, a significant increase of these regulatory T cells was found in irradiated CIA mice. Overall, our data suggest that low-dose gamma ray irradiation could attenuate CIA through suppression of pro-inflammatory cytokines and autoantibody production, and induction of regulatory T cells. PMID- 18413979 TI - Chromosome fragments have the potential to predict hyperthermia-induced radio sensitization in two different human tumor cell lines. AB - Cellular radiosensitivity, assessed by loss of clonogenicity, has been shown to correlate with the number of radiation-induced chromosomal aberrations. Also an increased radiosensitivity by hyperthermia has been shown to correlate with an increase in chromosomal aberrations. Therefore, determination of the number of chromosomal aberrations might be used as an assay to predict the radiosensitivity of tumors pre-treated with hyperthermia at clinically relevant temperatures. The use of premature chromosome condensation combined with fluorescent in situ hybridisation (PCC-FISH) has been shown to be clinically applicable. Therefore, the use of chromosomal aberrations as determined with PCC-FISH for the prediction of hyperthermia-induced radio-sensitization in human tumor cells was investigated. Confluent cultures of SW-1573 (human lung carcinoma) and RKO (human colorectal carcinoma) cells were treated with 1 h 41 degrees C or 43 degrees C hyperthermia prior to gamma-irradiation. Clonogenic cell survival and induction of chromosomal aberrations (unrejoined chromosomal fragments and translocations), by PCC-FISH, were studied at 24 h after treatment. Pre-treatment with hyperthermia at 41 degrees C for 1 h enhanced the radiosensitivity of RKO cells but not of SW-1573 cells. Increasing the temperature to 43 degrees C for 1 h enhanced the radiosensitivity of SW-1573 cells. When radio-sensitization was observed, a significant increase in the number of unrejoined chromosomal fragments was found but the frequency of translocations was not increased. Hyperthermia-induced radio-sensitization is correlated with an increase in unrejoined chromosomal fragments. This suggests that determination of the number of chromosomal fragments after hyperthermia and radiation treatment might be used for the prediction of combined treatment response in cancer patients. PMID- 18413980 TI - Histone H2AX phosphorylation independent of ATM after X-irradiation in mouse liver and kidney in situ. AB - Histone H2AX undergoes phosphorylation at Ser-139 (gamma-H2AX) rapidly in response to DNA double-strand breaks (DSBs) induced by ionizing radiation. The post-translational modification of H2AX plays a central role in responses to radiation, including the repair of DSBs. Although ataxia telangiectasia mutated (ATM) kinase phosphorylates Ser-139 of H2AX in vitro, the post-translational modification pattern and the modifier of H2AX in organs in vivo are not yet well understood. In this study, we detected phosphorylation of H2AX at Ser-139 in cells of the mouse ear, liver, and kidney after X-irradiation. Moreover, the phosphorylation of H2AX was regulated depending on not only the cell type, but also the organ type and the localization of a cell type in an organ. Following X irradiation, H2AX was phosphorylated in the liver and kidney of ATM gene knockout mice, suggesting that ATM kinase is not essential for phosphorylation of H2AX in these organs after X-irradiation in vivo. PMID- 18413981 TI - COMP-Ang1, angiopoietin-1 variant protects radiation-induced bone marrow damage in C57BL/6 mice. AB - Angiopoietin-1 (Ang1) is a vasculogenic factor which is signaled through the endothelial and bone marrow cell-specific, Tie2 receptor tyrosine kinase and has potential therapeutic applications for the induction of angiogenesis, enhancing endothelial cell survival, and preventing vascular leakage. In this study, we examined whether Ang1 directly exhibits bone marrow protection after ionizing radiation (IR) using an adenoviral vector of COMP-Ang1 (Ad-COMP-Ang1). This is a variant of Ang1 by replacement of the N-terminal portion of Ang1 with short coiled-coil domains of cartilage oligomeric matrix protein-Angiopoietin 1 (COMP Ang1) which are, long enough for oligomerization but short enough to avoid problems of aggregation and insolubility. A spleen colony assay after 4.5 Gy whole body radiation, indicated that COMP-Ang1 significantly increased the mean colony numbers. Both the decrease in bone marrow cellularity and increased TUNEL (Terminal deoxynucleotidyl Transferase Biotin-dUTP Nick End Labeling) positive cells produced by radiation in bone marrow were significantly inhibited by COMP Ang1 transfer. The expression of the ligands of Ang1 and Tie2 receptors were increased by radiation and, the COMP-Ang1 transfer potentiated this protein expression. Pre-treatment of Ang1 could be beneficial in protecting bone marrow from damage by radiation and COMP-Ang1 may be an effective alternative to native Ang1 for therapeutic purposes. PMID- 18413983 TI - Venous thromboembolism and prophylaxis in cancer patients. AB - Venous thromboembolism is a serious complication in patients with cancer. The seriousness of venous thromboembolism as a complication in cancer patients is becoming recognized as an important medical issue. Venous thromboembolism is a multifactorial disease associated with vascular endothelial damage, stasis of blood flow, and hypercoagulation. Preexisting morbidity, mutations of factor V Leiden or prothrombin 20210A, type of cancer, presence of metastases, use of central venous access, surgery, anesthesia, etc., increase the risk of venous thromboembolism. The patients with malignancies have a 7-fold increase in the risk of venous thromboembolism compared with individuals without cancer. Venous thromboembolism is the second most common cause of mortality in cancer patients. Venous thromboembolism is the most common cause of death at 30 days after surgery in patients undergoing surgery for cancer. Venous thromboembolism caused death in 46.3% of the cases after surgery for cancer. The Geneva prognostic index identified predictive factors for an adverse outcome, and the American College of Chest Physicians (ACCP) has suggested the guidelines for the prevention of venous tromboembolism in cancer patients. Cancer patients should receive appropriate venous thromboembolism prophylaxis. The methods used for venous thromboembolism prophylaxis are mechanical, pharmacological, or a combination of both. Well-timed thromboprophylaxis may protect patients from venous thromboembolism, early lethal outcome and even influence survival. PMID- 18413982 TI - Protection by polaprezinc against radiation-induced apoptosis in rat jejunal crypt cells. AB - Polaprezinc, an anti-ulcer drug, is a chelate compound consisting of zinc and L carnosine. Polaprezinc has been shown to prevent gastric mucosal injury. The anti ulcer effects of polaprezinc have been ascribed to its antioxidative property. The effect of polaprezinc on ionizing radiation-induced apoptosis was studied in the jejunal epithelial crypt cells of rats. Seven-to eight week-old Wistar rats, which were treated with 100 mg/kg of polaprezinc orally 1h before irradiation or 2% carboxymethyl cellulose sodium in controls, were exposed to whole body X-ray irradiation at 2 Gy. The number of apoptotic cells per jejunum crypt was counted in haematoxylin and eosin stained sections at 0-6 h after irradiation. TUNEL positive cells and immunopositive cells for active caspase-3 per crypt were also counted. Accumulation of p53, p21(WAF1/CIP1) and Bax expression in the jejunum after irradiation were examined by Western blot analyses. Polaprezinc treatment given prior to radiation resulted in a significant reduction in numbers of apoptotic cells, TUNEL positive cells and active caspase-3 immunopositive cells in jejunal crypt cells. Polaprezinc treatment resulted in decreases of p53 accumulation, p21(WAF1/CIP1) and Bax expression after irradiation. Polaprezinc has a protective effect against ionizing radiation induced apoptosis in rat jejunal crypt cells. PMID- 18413984 TI - [Prevalence of metabolic syndrome components in patients with acute coronary syndromes]. AB - OBJECTIVE: Many studies report that the components of the metabolic syndrome- arterial hypertension, abdominal obesity, diabetes mellitus, and atherogenic dyslipidemia--are associated with an increased risk of cardiovascular disease. We investigated the prevalence of different components of the metabolic syndrome and frequency of their combinations and acute hyperglycemia among patients with acute coronary syndromes. METHODS AND RESULTS: The study population consisted of 2756 patients (1670 men and 1086 women with a mean age of 63.3+/-11.3 years) with acute coronary syndromes: Q-wave myocardial infarction was present in 41.8% of patients; non-Q-wave MI, in 30.7%; and unstable angina pectoris, in 27.5%. The metabolic syndrome was found in 59.6% of the patients according to modified NCEP III guidelines. One component of the metabolic syndrome was found in 13.5% of patients; two, in 23.0%; and none, in 3.9%. Less than one-third (29.2%) of the patients had three components of the metabolic syndrome, and 30.4% of the patients had four or five components. Arterial hypertension and abdominal obesity were the most common components of the metabolic syndrome (82.2% and 65.8%, respectively). Nearly half of the patients had hypertriglyceridemia and decreased level of high-density lipoprotein cholesterol (55.0% and 51.1%, respectively), and 23.9% of patients had diabetes mellitus. Acute hyperglycemia (> or =6.1 mmol/L) without known diabetes mellitus was found in 38.1% of cases. The combination of arterial hypertension and abdominal obesity was reported in 57.8% of patients in the case of combinations of two-five metabolic syndrome components. CONCLUSION: More than half of patients with acute coronary syndromes had three or more components of the metabolic syndrome, and arterial hypertension and abdominal obesity were the most prevalent components of the metabolic syndrome. PMID- 18413985 TI - [The frequency of malignant disease in cytological group of suspected cancer (ultrasound-guided fine-needle aspiration biopsy of nonpalpable thyroid nodules)]. AB - BACKGROUND: Fine-needle aspiration biopsy is the most important procedure for differentiating benign thyroid nodules from malignant ones. Traditionally, findings of fine-needle aspiration biopsy are divided into four categories: nondiagnostic, benign lesions, suspected cancer, and malignant lesions. Group suspicious for cancer largely involves follicular neoplasms as well as lesions with cytological features of malignancy. OBJECTIVE: The purpose of this study was to establish the value of ultrasound-guided fine-needle aspiration biopsy in the diagnosis of nonpalpable thyroid cancer and to assess the frequency of malignant disease in the group of suspected cancer. PATIENTS AND METHODS: A total of 184 patients with nonpalpable thyroid nodules (less than 1.5 cm in diameter) were examined by means of ultrasound-guided fine-needle aspiration biopsy. Patients were included in the study based on sonographic findings implicating possible malignant nature of nodules. RESULTS. During 1997-2002, 204 ultrasound-guided fine-needle aspiration biopsies of thyroid nodules were performed; findings were nondiagnostic in 5.9% of cases. In 59.8% of cases, cytological examination revealed benign lesions; in 11.8%, suspected cancer; and in 22.5%, malignant lesions. Eighty-five patients underwent subsequent surgery with histological examination of specimens obtained. In 45 cases, cytological diagnosis of malignant or suspected thyroid cancer was confirmed by histological examination after surgery. CONCLUSION: We conclude that ultrasound-guided fine-needle aspiration biopsy has a high accuracy in the diagnostic evaluation of nonpalpable thyroid nodule with cytological features of malignancy. However, fine-needle aspiration biopsy is not effective diagnostic method for differentiating benign thyroid nodules from malignant ones in follicular neoplasm group. PMID- 18413986 TI - [Effect of smoking on neutrophil oxidative metabolism]. AB - Alterations in neutrophil function by tobacco products may play a central role in the pathogenesis of periodontal diseases and several smoking-related systemic diseases. The aim of the study was to evaluate the effect of smoking on neutrophil oxidative metabolism. MATERIALS AND METHODS: The study included 17 smoking men free of systemic diseases who were referred for treatment of various odontological diseases to outpatient department of Kaunas University of Medicine Hospital. The age of subjects varied from 22 to 43 years. All subjects answered the questions about smoking habits. Clinical examination included assessment of oral hygiene status according to the OHI-s index and periodontal status according to Russell and Ramfjord indices. To evaluate the oxidative metabolism of neutrophils, luminol- and liucigenin-dependent chemiluminescence and nitroblue tetrazolium test were used. RESULTS: After smoking, extracellular liucigenin dependent chemiluminescence response was higher as compared to the response before smoking, but total (intra- and extracellular) luminol-dependent chemiluminescence response was the same both before and after smoking. Exposure of neutrophils to smoking caused a significant increase in nitroblue tetrazolium reduction. CONCLUSION: The release of reactive oxygen species in neutrophils exposed to smoking may alter the pathogenic processes in periodontal diseases. PMID- 18413987 TI - [Microbiological and biochemical characteristics of inflammatory tissues in the periodontium]. AB - OBJECTIVE: To investigate bacterial populations in subgingival and supragingival plaque samples of patients with inflammatory periodontal diseases and activities of the lysosomal enzymes--lysozyme, alkaline phosphatase, and beta-glucuronidase- in peripheral venous blood, in gingival crevicular fluid, and mixed nonstimulated saliva. METHODS AND MATERIALS: The study included 60 patients with inflammatory periodontal diseases without any internal pathology and 24 periodontally healthy subjects. Molecular genetic assay (Micro-IDent plus, Germany) for complex identification of additional six periodontopathic bacteria was applied. The activity of lysozyme was determined turbidimetrically, the activity of alkaline phosphatase--spectrophotometrically with a "Monarch" biochemical analyzer, the activity beta-glucuronidase--according to the method described by Mead et al. and modified by Strachunskii. RESULTS: A statistically significant association between clinical and bacteriological data was found in the following cases: gingival bleeding in the presence of Eubacterium nodatum, Eikenella corrodens, Capnocytophaga spp. (P<0.01); pathological periodontal pockets in the presence of Peptostreptococcus micros (alpha< or =0.05 and beta< or =0.2), Fusobacterium nucleatum (alpha< or =0.05 and beta< or =0.2), Campylobacter rectus (alpha< or =0.05 and beta< or =0.2), and Capnocytophaga spp. (P<0.05); and satisfactory oral hygiene in the presence of all microorganisms investigated (P<0.05). The activity of lysozyme in gingival crevicular fluid and mixed nonstimulated saliva indicates the severity of periodontal inflammation. Based on clinical data, in assessing the amount of lysozyme in mixed nonstimulated saliva, sensitivity and specificity of 100% was found. Increased activities of lysozyme, alkaline phosphatase, and beta-glucuronidase were found in peripheral venous blood of patients with inflammatory periodontal disease as compared to control group. CONCLUSIONS: The main principles of the treatment of periodontal inflammatory diseases should be based on microorganism elimination, creation of individual treatment means affecting microflora in the mouth and immune system of macroorganisms. PMID- 18413988 TI - Gastroesophageal reflux disease after Helicobacter pylori eradication in gastric ulcer patients: a one-year follow-up study. AB - The aim of this study was to evaluate the course of gastroesophageal reflux disease in gastric ulcer patients after successful Helicobacter pylori eradication (group A), in patients with persistent infection after attempt to eradicate Helicobacter pylori (group B), and in control group without Helicobacter pylori eradication treatment (group C). MATERIALS AND METHODS: Gastric ulcer patients (n=88) were assigned either to the group receiving Helicobacter pylori eradication treatment (54 patients) or to the control group (34 patients; omeprazole treatment for 4 weeks) and were followed up for 1-year or until gastric ulcer relapsed. Gastroesophageal reflux disease was diagnosed in patients who had erosive esophagitis and/or without esophagitis if they experienced heartburn and/or regurgitation at least 2 times a week and it was associated with impairment of daily activities (Genval consensus). RESULTS: The study was completed by 69 patients: 25 in group A, 19 in group B, and 25 in group C. At the beginning and at the end of the follow-up, gastroesophageal reflux disease was diagnosed in 10 (40%) and 9 (36%) group A patients, respectively (P>0.05); in 12 (63%) and 8 (42%) group B patients, respectively (P>0.05); and in 9 (36%) and 5 (20%) group C patients, respectively (P>0.05). At the beginning and at the end of the follow-up, reflux esophagitis was found in 3 (12%) and 5 (20%) group A patients, respectively (P>0.05); in 5 (26%) and 5 (26%) group B patients (P>0.05); in 4 (16%) and 3 (12%) group C patients (P>0.05). CONCLUSION: There was no statistically significant difference regarding the development of gastroesophageal reflux disease in gastric ulcer patients after Helicobacter pylori eradication, in the patients with persistent infection after attempt to eradicate, and in the control group without Helicobacter pylori eradication treatment. PMID- 18413989 TI - Evaluation of activity and effectiveness of occupational therapy in stroke patients at the early stage of rehabilitation. AB - In Lithuania, the stroke is not only medical, but social issue as well, since only 20% of patients suffering from stroke remain active at work. Yearly stroke incidence in Lithuania is 7000-8000 cases. The most common outcome of stroke is unilateral paralysis (hemiplegia) followed by disorders of coordination, balance, and movements. Due to dysfunctions of movements, self-care, cognition, behavior, and communication, some part of stroke patients remains disabled. They need assistance and care provided by other people. Occupational therapy, which is part of rehabilitation of patients after stroke, is directed to independence training. There are scarce data related to effectiveness of occupational therapy depending on motor, cognitive, and psychosocial dysfunctions. Goals of study were to estimate effectiveness of occupational therapy at the early stage of rehabilitation depending on type of stroke, localization of brain injury, grade of lesion, age, and gender, to identify factors influencing effectiveness of occupational therapy, and to estimate their positive predictive value. The study included 106 patients at the early stage of rehabilitation, who were admitted to Department of Neurorehabilitation after stabilization of clinical condition from Departments of Neurology and Neurosurgery (mean duration of 14+/-2 days after stroke). The program of occupational therapy was not fulfilled by 6 patients: 2 patients were transferred to Nursing Hospital due to severe condition, and 4 patients were discharged prematurely and continued rehabilitation in outpatient setting. Hence, study population consisted of 100 subjects (47 men and 53 women) who were diagnosed with stroke (ischemic or hemorrhagic). Patient's functional status and disorders of activities were evaluated using Barthel Index and Functional Independence Measure. Complexes of occupational therapy were adjusted according to examination of patient's disorders of activities, age, grade of lesion, other diseases, and complications during rehabilitation process. Effectiveness of occupational therapy in patients after stroke at then early stage of rehabilitation was influenced by gender, age, degree of lesion, type of stroke. Better effectiveness of occupational therapy was observed in men (P<0.05), persons younger than 59 years (P<0.05), persons with hemiparesis (P<0.05) and hemorrhagic stroke (P<0.05). Low effectiveness in stroke patients at the early stage of rehabilitation was influenced by neglect (P<0.05), hemiplegia (P<0.001), and older age of patient (P<0.05). Application of individualized occupational therapy complexes for stroke patients at the early stage of rehabilitation with consideration of motoric and cognitive-psychosocial disorders, significantly improves recovery of impaired functions, though occupational therapy should be continued in later rehabilitation stages, since patients after early stage of rehabilitation still have limited independence in daily activities. PMID- 18413990 TI - [Cooling makes recovery of muscle faster after eccentric-concentric than concentric exercise]. AB - The aim of the study was to establish the influence of muscle cooling on muscle recovery after concentric and eccentric-concentric exercise. Healthy untrained males (n=40) took part in this study. During the first experiment, subjects performed concentric and during the second--eccentric-concentric exercise. During both experiments, the subjects were divided into the groups. In the first group, the muscles of the lower limbs were cooled down after the physical load, while the muscles of the subjects of the second group were not cooled. The rectal, skin, and muscle temperature was repeatedly measured immediately after physical load and after cooling of the muscle. Before exercise and after 2 min and 4, 8, 24, 48, 72 hours after performing exercise, quadriceps muscle strength, generated by electrical stimulation at frequencies of 20 Hz and 50 Hz, and maximal voluntary contraction force were registered. Serum creatine kinase levels were measured before and 24 hours after exercise. In addition, the subjects subjectively rated their muscle pain on a 10-point scale 24, 48, 72 hours after exercise. The results show that cooling applied to muscles after concentric and eccentric-concentric exercise affected the indicators of muscle damage--the activity of creatine kinase was decreased and muscle strength recovered faster. The cooling effect is greater when it is applied after eccentric-concentric exercise. PMID- 18413991 TI - [School bullying and its association with health and lifestyle among schoolchildren]. AB - The purpose of the study was to assess the association of bullying and being bullied with indicators of subjective health and problem behaviors among schoolchildren aged 11-15 years. MATERIAL AND METHODS: The data of the anonymous survey of 5645 filth-, seventh-, and ninth-grade students (aged 11, 13, and 15 years, respectively), conducted in the spring of 2002, were analyzed. The students completed the World Health Organization's Health Behavior in School-aged Children questionnaire that included self-report of involvement in bullying and being bullied by others as well as subjective health and well-being estimates, health complains (headache, stomachache, back pain, anxiety, etc.), and problem behaviors (smoking, alcohol and drug use, suicidal ideation). The response rate was 95%. RESULTS: More than half (52.3%) of students were involved in bullying process at least two times per month: 17.9% were involved as bullies ("victims"), 18.3% were bullied ("aggressors"), and 16.1% bullied others and were bullied themselves. A significant association between experiencing bullying and adverse health outcomes was found. Perpetrating bullying increased the odds of smoking, alcohol and drug use. Perpetrating and/or experiencing bullying increased the risk of high suicidal ideation that had cumulative effect. The established associations varied between genders and groups of students defined as "victims," "aggressors," and "victims/aggressors." CONCLUSIONS: In Lithuania, school bullying is extremely prevalent and is associated with health disorders, poorer well-being, and problem behavior of schoolchildren. Urgent antibullying efforts, including both research and preventive measures, are needed in order to deal with this social phenomenon. PMID- 18413992 TI - Factors associated with poor sleep and health-related quality of life. AB - OBJECTIVE: The aim of the study was to establish factors associated with poor sleep and to assess the relationship between self-evaluated sleep quality and health-related quality of life. MATERIAL AND METHODS: Sleep complaints were evaluated using Basic Nordic Sleep Questionnaire, and health-related quality of life was assessed by SF-36. Subjective data about sleep quality were obtained from 1602 randomly selected persons: 600 males and 1002 females, aged 35-74 years. SF-36 was filled in by 1016 persons: 379 males and 637 females. Health status was evaluated by Perceived Health Questionnaire. The odds ratios of poor sleep were calculated using binary logistic regression analysis. RESULTS: Among males poor self-evaluated health, frequent stress events, regular nighttime awakenings, and sleep latency period longer than 15 min in workdays were significant predictors of poor sleep. Among females, duration of sleep shorter than 7 h, frequent stress events, poor self-evaluated health, sleep latency period longer than 15 min in workdays, and regular nighttime awakenings predicted poor sleep. Poor sleepers, as compared with good ones, had poorer health-related quality of life. CONCLUSIONS: Poor perceived health, frequent stress events, regular nighttime awakenings, and sleep latency period longer than 15 min were indicated as significant predictors of poor sleep. Poor sleep worsened health related quality of life in all domains of SF-36. PMID- 18413993 TI - [Aortic dissection]. AB - Aortic dissection is an acute lesion of the aortic wall accompanied by separation of the media due to rupture or intramural hematoma. The incidence rate of aortic dissection is 5 to 30 cases per million people a year. Acute aortic dissection is a highly lethal cardiovascular emergency with an incidence of 2000 new cases per year in the United States and 3000 in Europe. The mortality rate of aortic dissection is 3.2/100,000 per year. In case of sudden death of nonhospitalized patients, aortic dissection was proved in 1.5% of necropsy cases. Most of patients die within 48 hours after admission or 1.4% per each hour. The main clinical manifestations of aortic dissection are acute myocardial infarction, stroke, pulmonary embolism, acute heart failure, acute pancreatitis, mesenteries thrombosis, which mislead the physician. The main measure, which might reduce the mortality, is early diagnosis of aortic dissection. The standard diagnosis is based on clinical symptoms and verification by instrumental (imaging) methods. An alternative mean for diagnosis of aortic dissection might be the determination of concentration of smooth muscle myosin heavy chain protein in blood serum, the peak of which is found after 3 hours after the onset of pain. Normal value of smooth muscle myosin heavy chain protein concentration is 2.5 microg/L, while in case of aortic dissection it exceeds 22.4 microg/L. This diagnostic method has not been introduced in Lithuania yet. PMID- 18413994 TI - Flavanone 3beta-hydroxylases from rice: key enzymes for favonol and anthocyanin biosynthesis. AB - Flavanone 3beta-hydroxylases (F3H) are key enzymes in the synthesis of flavonol and anthocyanin. In this study, three F3H cDNAs from Oryza sativa (OsF3H-1 approximately 3) were cloned by RT-PCR and expressed in E. coli as gluthatione S transferase (GST) fusion proteins. The purified recombinant OsF3Hs used flavanone, naringenin and eriodictyol as substrates. The reaction products with naringen and eriodictyol were determined by nuclear magnetic resonance spectroscopy to be dihydrokaempferol and taxifolin, respectively. OsF3H-1 had the highest enzymatic activity whereas the overall expression of OsF3H-2 was highest in all tissues except seeds. Flavanone 3beta-hydroxylase could be a useful target for flavonoid metabolic engineering in rice. PMID- 18413995 TI - Bone marrow-derived side population cells are capable of functional cardiomyogenic differentiation. AB - It has been reported that bone marrow (BM)-side population (SP) cells, with hematopoietic stem cell activity, can transdifferentiate into cardiomyocytes and contribute to myocardial repair. However, this has been questioned by recent studies showing that hematopoietic stem cells (HSCs) adopt a hematopoietic cell lineage in the ischemic myocardium. The present study was designed to investigate whether BM-SP cells can in fact transdifferentiate into functional cardiomyocytes. Phenotypically, BM-SP cells were 19.59% (+/-)9.00 CD14+, 8.22% (+/-)2.72 CD34+, 92.93% (+/-)2.68 CD44+, 91.86% (+/-)4.07 CD45+, 28.48% (+/-)2.24 c-kit+, 71.09% (+/-)3.67 Sca-1+. Expression of endothelial cell markers (CD31, Flk-1, Tie-2 and VEGF-A) was higher in BM-SP cells than whole BM cells. After five days of co-culture with neonatal cardiomyocytes, 7.2% (+/-)1.2 of the BM-SP cells expressed sarcomeric alpha-actinin as measured by flow cytometry. Moreover, BM-SP cells co-cultured on neonatal cardiomyocytes fixed to inhibit cell fusion also expressed sarcomeric alpha-actinin. The co-cultured BM-SP cells showed neonatal cardiomyocyte-like action potentials of relatively long duration and shallow resting membrane potential. They also generated calcium transients with amplitude and duration similar to those of neonatal cardiomyocytes. These results show that BM-SP cells are capable of functional cardiomyogenic differentiation when co-cultured with neonatal cardiomyocytes. PMID- 18413996 TI - Heat stress causes aberrant DNA methylation of H19 and Igf-2r in mouse blastocysts. AB - To gain a better understanding of the methylation imprinting changes associated with heat stress in early development, we used bisulfite sequencing and bisulfite restriction analysis to examine the DNA methylation status of imprinted genes in early embryos (blastocysts). The paternal imprinted genes, H19 and Igf-2r, had lower methylation levels in heat-stressed embryos than in control embryos, whereas the maternal imprinted genes, Peg3 and Peg1, had similar methylation pattern in heat-stressed embryos and in control embryos. Our results indicate that heat stress may induce aberrant methylation imprinting, which results in developmental failure of mouse embryos, and that the effects of heat shock on methylation imprinting may be gene-specific. PMID- 18413997 TI - Molecular changes in remote tissues induced by electro-acupuncture stimulation at acupoint ST36. AB - To investigate the effects of electro-acupuncture (EA) treatment on regions remote from the application, we measured cellular, enzymatic, and transcriptional activities in various internal tissues of healthy rats. The EA was applied to the well-identified acupoint ST36 of the leg. After application, we measured the activity of natural killer cells in the spleen, gene expression in the hypothalamus, and the activities of antioxidative enzymes in the hypothalamus, liver and red blood cells. The EA treatment increased natural killer cell activity in the spleen by approximately 44%. It also induced genes related to pain, including 5-Hydroxytryptamine (serotonin) receptor 3a (Htr3a) and Endothelin receptor type B (Ednrb) in the hypothalamus, and increased the activity of superoxide dismutase in the hypothalamus, liver, and red blood cells. These findings indicate that EA mediates its effects through changes in cellular activity, gene expression, and enzymatic activity in multiple remote tissues. The sum of these alterations may explain the beneficial effects of EA. PMID- 18413998 TI - Genetic characteristics of 207 microsatellite markers in the Korean population and in other Asian populations. AB - Microsatellites, short tandem repeats, are useful markers for genetic analysis because of their high frequency of occurrence over the genome, high information content due to variable repeat lengths, and ease of typing. To establish a panel of microsatellite markers useful for genetic studies of the Korean population, the allele frequencies and heterozygosities of 207 microsatellite markers in 119 unrelated Korean, Indian and Pakistani individuals were compared. The average heterozygosity of the Korean population was 0.71, similar to that of the Indian and Pakistani populations. More than 80% of the markers showed heterozygosity of over 0.6 and were valuable as genetic markers for genome-wide screening for disease susceptibility loci in these populations. To identify the allelic distributions of the multilocus genetic data from these microsatellite markers, the population structures were assessed by clustering. These markers supported, with the most probability, three clustering groups corresponding to the three geographical populations. When we assumed only two hypothetical clusters (K), the Korean population was separate from the others, suggesting a relatively deep divergence of the Korean population. The present 207 microsatellite markers appear to reflect the historical and geographical origins of the different populations as well as displaying a similar degree of variation to that seen in previously published genetic data. Thus, these markers will be useful as a reference for human genetic studies on Asians. PMID- 18413999 TI - Arabidopsis histidine-containing phosphotransfer factor 4 (AHP4) negatively regulates secondary wall thickening of the anther endothecium during flowering. AB - Cytokinins are essential hormones in plant development. Arabidopsis histidine containing phosphotransfer proteins (AHPs) are mediators in a multistep phosphorelay pathway for cytokinin signaling. The exact role of AHP4 has not been elucidated. In this study, we demonstrated young flower-specific expression of AHP4, and compared AHP4-overexpressing (Ox) trangenic Arabidopsis lines and an ahp4 knock-out line. AHP4-Ox plants had reduced fertility due to a lack of secondary cell wall thickening in the anther endothecium and inhibition of IRREGURAR XYLEMs (IRXs) expression in young flowers. Conversely, ahp4 anthers had more lignified anther walls than the wild type, and increased IRXs expression. Our study indicates that AHP4 negatively regulates thickening of the secondary cell wall of the anther endothecium, and provides new insight into the role of cytokinins in formation of secondary cell walls via the action of AHP4. PMID- 18414000 TI - Growth retardation and death of rice plants irradiated with carbon ion beams is preceded by very early dose- and time-dependent gene expression changes. AB - The carbon-ion beam (CIB) generated by the heavy-ion medical accelerator in Chiba (HIMAC) was targeted to 7-day-old rice. Physiological parameters such as growth, and gene expression profiles were examined immediately after CIB irradiation. Dose-dependent growth suppression was seen three days post-irradiation (PI), and all the irradiated plants died by 15 days PI. Microarray (Agilent rice 22K) analysis of the plants immediately after irradiation (iai) revealed effects on gene expression at 270 Gy; 353 genes were up-regulated and 87 down-regulated. Exactly the same set of genes was affected at 90 Gy. Among the highly induced genes were genes involved in information storage and processing, cellular processes and signaling, and metabolism. RT-PCR analysis confirmed the microarray data. PMID- 18414001 TI - Identification and characterization of single nucleotide polymorphisms of SLC22A11 (hOAT4) in Korean women osteoporosis patients. AB - Single nucleotide polymorphisms (SNPs) are the most common form of human genetic variation. Non-synonymous SNPs (nsSNPs) change an amino acid. Organic anion transporters (OATs) play an important role in eliminating or reabsorbing endogenous and exogenous organic anionic compounds. Among OATs, hOAT4 mediates high affinity transport of estrone sulfate and dehydroepiandrosterone sulfate. The rapid bone loss that occurs in post-menopausal women is mainly due to a net decrease of estrogen. In the present study we searched for SNPs within the exon regions of hOAT4 in Korean women osteoporosis patients. Fifty healthy subjects and 50 subjects with osteoporosis were screened for genetic polymorphism in the coding region of SLC22A11 (hOAT4) using GC-clamp PCR and denaturing gradient gel electrophoresis (DGGE). We found three SNPs in the hOAT4 gene. Two were in the osteoporosis group (C483A and G832A) and one in the normal group (C847T). One of the SNPs, G832A, is an nsSNP that changes the 278th amino acid from glutamic acid to lysine (E278K). Uptake of [3H] estrone sulfate by oocytes injected with the hOAT4 E278K mutant was reduced compared with wild-type hOAT4. Km values for wild type and E278K were 0.7 microM and 1.2 microM, and Vmax values were 1.8 and 0.47 pmol/oocyte/h, respectively. The present study demonstrates that hOAT4 variants can causing inter-individual variation in anionic drug uptake and, therefore, could be used as markers for certain diseases including osteoporosis. PMID- 18414002 TI - Functional equivalence of translation factor eIF5B from Candida albicans and Saccharomyces cerevisiae. AB - Eukaryotic translation initiation factor 5B (eIF5B) plays a role in recognition of the AUG codon in conjunction with translation factor eIF2, and promotes joining of the 60S ribosomal subunit. To see whether the eIF5B proteins of other organisms function in Saccharomyces cerevisiae, we cloned the corresponding genes from Oryza sativa, Arabidopsis thaliana, Aspergillus nidulans and Candida albican and expressed them under the control of the galactose-inducible GAL promoter in the fun12Delta strain of Saccharomyces cerevisiae. Expression of Candida albicans eIF5B complemented the slow-growth phenotype of the fun12Delta strain, but that of Aspergillus nidulance did not, despite the fact that its protein was expressed better than that of Candida albicans. The Arabidopsis thaliana protein was also not functional in Saccharomyces. These results reveal that the eIF5B in Candida albicans has a close functional relationship with that of Sacharomyces cerevisiae, as also shown by a phylogenetic analysis based on the amino acid sequences of the eIF5Bs. PMID- 18414003 TI - Stage-specific expression of ankyrin and SOCS box protein-4 (Asb-4) during spermatogenesis. AB - Members of the large family of Asb proteins are ubiquitously expressed in mammalian tissues; however, the roles of individual Asb and their function in the developmental testes have not been reported. In this report, we isolated a murine Asb4 from mouse testis. Northern blot analysis revealed that mAsb-4 was expressed only in testes and produced in a stage-specific manner during spermatogenesis. It was expressed in murine testes beginning in the fourth week after birth and extending into adulthood. Pachytene spermatocytes had the highest level of expression. Interestingly, the human homologue of mAsb-4, ASB-4 (hASB-4) was also expressed in human testis. These results suggest that ASB-4 plays pivotal roles in mammalian testis development and spermatogenesis. PMID- 18414004 TI - Acrolein with an alpha, beta-unsaturated carbonyl group inhibits LPS-induced homodimerization of toll-like receptor 4. AB - Acrolein is a highly electrophilic alpha,beta-unsaturated aldehyde present in a number of environmental sources, especially cigarette smoke. It reacts strongly with the thiol groups of cysteine residues by Michael addition and has been reported to inhibit nuclear factor-kappaB (NF-kappaB) activation by lipopolysaccharide (LPS). The mechanism by which it inhibits NF-kappaB is not clear. Toll-like receptors (TLRs) play a key role in sensing microbial components and inducing innate immune responses, and LPS-induced dimerization of TLR4 is required for activation of downstream signaling pathways. Thus, dimerization of TLR4 may be one of the first events involved in activating TLR4-mediated signaling pathways. Stimulation of TLR4 by LPS activates both myeloid differential factor 88 (MyD88)- and TIR domain-containing adapter inducing IFNbeta(TRIF)-dependent signaling pathways leading to activation of NF-kappaB and IFN-regulatory factor 3 (IRF3). Acrolein inhibited NF-kappaB and IRF3 activation by LPS, but it did not inhibit NF-kappaB or IRF3 activation by MyD88, inhibitor kappaB kinase (IKK)beta, TRIF, or TNF-receptor-associated factor family member associated NF-kappaB activator (TANK)-binding kinase 1 (TBK1). Acrolein inhibited LPS-induced dimerization of TLR4, which resulted in the down-regulation of NF kappaB and IRF3 activation. These results suggest that activation of TLRs and subsequent immune/inflammatory responses induced by endogenous molecules or chronic infection can be modulated by certain chemicals with a structural motif that enables Michael addition. PMID- 18414005 TI - Accumulation of flavonols in response to ultraviolet-B irradiation in soybean is related to induction of flavanone 3-beta-hydroxylase and flavonol synthase. AB - There are several branch points in the flavonoid synthesis pathway starting from chalcone. Among them, the hydroxylation of flavanone is a key step leading to flavonol and anthocyanin. The flavanone 3-beta-hydroxylase (GmF3H) gene was cloned from soybean (Glycine max cultivar Sinpaldal) and shown to convert eriodictyol and naringenin into taxifolin and dihydrokaempferol, respectively. The major flavonoids in this soybean cultivar were found by LC-MS/MS to be kamepferol O-triglycosides and O-diglycosides. Expression of GmF3H and flavonol synthase (GmFLS) was induced by ultraviolet-B (UV-B) irradiation and their expression stimulated accumulation of kaempferol glycones. Thus, GmF3H and GmFLS appear to be key enzymes in the biosynthesis of the UV-protectant, kaempferol. PMID- 18414006 TI - Arginine deiminase enhances MCF-7 cell radiosensitivity by inducing changes in the expression of cell cycle-related proteins. AB - After successful clinical application, arginine deiminase (ADI) has been proposed to be a new cancer therapeutic. In the present study, we examined the effect of ADI in combination with ionizing radiation (IR) on MCF-7 cell growth and clonogenic cell death. Cell growth was inhibited by IR in a dose-dependent manner and ADI enhanced the radiosensitivity. ADI itself did not suppress the growth of MCF-7 cells due to the high level of expression of argininosuccinate synthetase (ASS), which convert citrulline, a product of arginine degradation by ADI, to arginine. Previously, it was suggested that ammonia, another product of arginine degradation by ADI, is the main cause of the growth inhibition of irradiated hepatoma cells contaminated with ADI-expressing mycoplasma [van Rijn et al. (2003)]. However, we found that ammonia is not the only factor that enhances radiosensitivity, as enhancement was also observed in the absence of ammonia. In order to identify the enhancing effect, levels of ASS and proteins related to the cell cycle were examined. ASS was unchanged by ADI plus IR, but p21 (a CDK inhibitor) was upregulated and c-Myc downregulated. These findings indicate that changes in the expressions of cell cycle proteins are involved in the enhancement of radiosensitivity by ADI. We suggest that ADI is a potential adjunct to cancer therapy. PMID- 18414007 TI - Repression of transcriptional activity of estrogen receptor alpha by a Cullin3/SPOP ubiquitin E3 ligase complex. AB - The role of SPOP in the ubiquitination of ER alpha by the Cullin3-based E3 ubiquitin ligase complex was investigated. We showed that the N-terminal region of SPOP containing the MATH domain interacts with the AF-2 domain of ER alpha in cultured human embryonic 293 cells. SPOP was required for coimmunoprecipitation of ER alpha; with Cullin3. This is the first report of the essential role of SPOP in ERalpha ubiquitination by the Cullin3-based E3 ubiquitin ligase complex. We also demonstrated repression of the transactivation capability of ER alpha; in cultured mammalian cells. PMID- 18414008 TI - Clustering approaches to identifying gene expression patterns from DNA microarray data. AB - The analysis of microarray data is essential for large amounts of gene expression data. In this review we focus on clustering techniques. The biological rationale for this approach is the fact that many co-expressed genes are co-regulated, and identifying co-expressed genes could aid in functional annotation of novel genes, de novo identification of transcription factor binding sites and elucidation of complex biological pathways. Co-expressed genes are usually identified in microarray experiments by clustering techniques. There are many such methods, and the results obtained even for the same datasets may vary considerably depending on the algorithms and metrics for dissimilarity measures used, as well as on user selectable parameters such as desired number of clusters and initial values. Therefore, biologists who want to interpret microarray data should be aware of the weakness and strengths of the clustering methods used. In this review, we survey the basic principles of clustering of DNA microarray data from crisp clustering algorithms such as hierarchical clustering, K-means and self organizing maps, to complex clustering algorithms like fuzzy clustering. PMID- 18414009 TI - Expressed sequence tag analysis of Antarctic hairgrass Deschampsia antarctica from King George Island, Antarctica. AB - Deschampsia antarctica is the only monocot that thrives in the tough conditions of the Antarctic region. It is an invaluable resource for the identification of genes associated with tolerance to various environmental pressures. In order to identify genes that are differentially regulated between greenhouse-grown and Antarctic field-grown plants, we initiated a detailed gene expression analysis. Antarctic plants were collected and greenhouse plants served as controls. Two different cDNA libraries were constructed with these plants. A total of 2,112 cDNA clones was sequenced and grouped into 1,199 unigene clusters consisting of 243 consensus and 956 singleton sequences. Using similarity searches against several public databases, we constructed a functional classification of the ESTs into categories such as genes related to responses to stimuli, as well as photosynthesis and metabolism. Real-time PCR analysis of various stress responsive genes revealed different patterns of regulation in the different environments, suggesting that these genes are involved in responses to specific environmental factors. PMID- 18414010 TI - N-acetylphytosphingosine enhances the radiosensitivity of lung cancer cell line NCI-H460. AB - Ceramides are well-known second messengers that induce apoptosis in various kinds of cancer cells, and their effects are closely related to radiation sensitivity. Phytoceramides, the yeast counterparts of the mammalian ceramides, are also reported to induce apoptosis. We investigated the effect of a novel ceramide derivative, N-acetylphytosphingosine (NAPS), on the radiosensitivity of NCI-H460 human lung carcinoma cells and its differential cytotoxicity in tumor and normal cells. The combination of NAPS with radiation significantly increased clonogenic cell death and caspase-dependent apoptosis. The combined treatment greatly increased Bax expression and Bid cleavage, but not Bcl-2 expression. However, there was no effect on radiosensitivity and apoptosis in BEAS2B cells, which derive from normal human bronchial epithelium. Cell proliferation and DNA synthesis were significantly inhibited by NAPS in both NCI-H460 and BEAS2B cells, but only the BEAS2B cells recovered by 48h after removal of the NAPS. Furthermore, the NCI-H460 cells underwent more DNA fragmentation than the BEAS2B cells in response to NAPS. Our results indicate that NAPS may be a potential radiosensitizing agent with differential effects on tumor vs. normal cells. PMID- 18414011 TI - Development of a sequence characteristic amplified region marker linked to the L4 locus conferring broad spectrum resistance to tobamoviruses in pepper plants. AB - To develop molecular markers linked to the L4 locus conferring resistance to tobamovirus pathotypes in pepper plants, we performed AFLP with 512 primer combinations for susceptible (S pool) and resistant (R pool) DNA bulks against pathotype 1.2 of pepper mild mottle virus. Each bulk was made by pooling the DNA of five homozygous individuals from a T10 population, which was a near-isogenic BC4F2 generation for the L4 locus. A total of 19 primer pairs produced scorable bands in the R pool. Further screening with these primer pairs was done on DNA bulks from T102, a BC10F2 derived from T10 by back crossing. Three AFLP markers were finally selected and designated L4-a, L4-b and L4-c. L4-a and L4-c each underwent one recombination event, whereas no recombination for L4-b was seen in 20 individuals of each DNA bulk. Linkage analysis of these markers in 112 F2 T102 individuals showed that they were each within 2.5 cM of the L4 locus. L4-b was successfully converted into a simple 340-bp SCAR marker, designated L4SC340, which mapped 1.8 cM from the L4 locus in T102 and 0.9 cM in another BC10F2 population, T101. We believe that this newly characterized marker will improve selection of tobamovirus resistance in pepper plants by reducing breeding cost and time. PMID- 18414012 TI - Attenuated neuropathic pain in Cav3.1 null mice. AB - To assess the role of alpha(1G) T-type Ca2+ channels in neuropathic pain after L5 spinal nerve ligation, we examined behavioral pain susceptibility in mice lacking CaV3.1 (alpha1G(-/-)), the gene encoding the pore-forming units of these channels. Reduced spontaneous pain responses and an increased threshold for paw withdrawal in response to mechanical stimulation were observed in these mice. The alpha1G(-/-) mice also showed attenuated thermal hyperalgesia in response to both low-(IR30) and high-intensity (IR60) infrared stimulation. Our results reveal the importance of alpha(1G) T-type Ca2+ channels in the development of neuropathic pain, and suggest that selective modulation of alpha1G subtype channels may provide a novel approach to the treatment of allodynia and hyperalgesia. PMID- 18414013 TI - Metabolic engineering of indole glucosinolates in Chinese cabbage plants by expression of Arabidopsis CYP79B2, CYP79B3, and CYP83B1. AB - Indole glucosinolates (IG) play important roles in plant defense, plant-insect interactions, and stress responses in plants. In an attempt to metabolically engineer the IG pathway flux in Chinese cabbage, three important Arabidopsis cDNAs, CYP79B2, CYP79B3, and CYP83B1, were introduced into Chinese cabbage by Agrobacterium-mediated transformation. Overexpression of CYP79B3 or CYP83B1 did not affect IG accumulation levels, and overexpression of CYP79B2 or CYP79B3 prevented the transformed callus from being regenerated, displaying the phenotype of indole-3-acetic acid (IAA) overproduction. However, when CYP83B1 was overexpressed together with CYP79B2 and/or CYP79B3, the transformed calli were regenerated into whole plants that accumulated higher levels of glucobrassicin, 4 hydroxy glucobrassicin, and 4-methoxy glu-cobrassicin than wild-type controls. This result suggests that the flux in Chinese cabbage is predominantly channeled into IAA biosynthesis so that coordinate expression of the two consecutive enzymes is needed to divert the flux into IG biosynthesis. With regard to IG accumulation, overexpression of all three cDNAs was no better than overexpression of the two cDNAs. The content of neoglucobrassicin remained unchanged in all transgenic plants. Although glucobrassicin was most directly affected by overexpression of the transgenes, elevated levels of the parent IG, glucobrassicin, were not always accompanied by increases in 4-hydroxy and 4 methoxy glucobrassicin. However, one transgenic line producing about 8-fold increased glucobrassicin also accumulated at least 2.5 fold more 4-hydroxy and 4 methoxy glucobrassicin. This implies that a large glucobrassicin pool exceeding some threshold level drives the flux into the side chain modification pathway. Aliphatic glucosinolate content was not affected in any of the transgenic plants. PMID- 18414014 TI - Marker production by PCR amplification with primer pairs from conserved sequences of WRKY genes in chili pepper. AB - Despite increasing awareness of the importance of WRKY genes in plant defense signaling, the locations of these genes in the Capsicum genome have not been established. To develop WRKY-based markers, primer sequences were deduced from the conserved sequences of the DNA binding motif within the WRKY domains of tomato and pepper genes. These primers were derived from upstream and downstream parts of the conserved sequences of the three WRKY groups. Six primer combinations of each WRKY group were tested for polymorphisms between the mapping parents, C. annuum 'CM334' and C. annuum 'Chilsungcho'. DNA fragments amplified by primer pairs deduced from WRKY Group II genes revealed high levels of polymorphism. Using 32 primer pairs to amplify upstream and downstream parts of the WRKY domain of WRKY group II genes, 60 polymorphic bands were detected. Polymorphisms were not detected with primer pairs from downstream parts of WRKY group II genes. Half of these primers were subjected to F2 genotyping to construct a linkage map. Thirty of 41 markers were located evenly spaced on 20 of the 28 linkage groups, without clustering. This linkage map also consisted of 199 AFLP and 26 SSR markers. This WRKY-based marker system is a rapid and simple method for generating sequence-specific markers for plant gene families. PMID- 18414015 TI - C-FLIP promotes the motility of cancer cells by activating FAK and ERK, and increasing MMP-9 expression. AB - We examined the role of c-FLIP in the motility of HeLa cells. A small interfering RNA (siRNA) directed against c-FLIP inhibited the adhesion and motility of the cells without affecting their growth rate. The long form of c-FLIP (c-FLIPL), but not the short form (c-FLIPS), enhanced adhesion and motility. Downregulation of c FLIPL with siRNA decreased phosphorylation of FAK and ERK, while overexpression of c-FLIPL increased their phosphorylation. Overexpression of FAK activated ERK, and enhanced the motility of HeLa cells. FRNK, an inhibitory fragment of FAK, inhibited ERK and decreased motility. Inhibition of ERK also significantly suppressed c-FLIPL-promoted motility. Inhibition of ROCK by Y27632 suppressed the c-FLIPL-promoted motility by reducing phosphorylation of FAK and ERK. Overexpression of c-FLIPL increased the expression and secretion of MMP-9, and inhibition of MMP-9 by Ilomastat reduced c-FLIPL- promoted cell motility. A caspase-like domain (amino acids 222-376) was found to be necessary for the c FLIPL-promoted cell motility. We conclude that c-FLIPL promotes the motility of HeLa cells by activating FAK and ERK, and increasing MMP-9 expression. PMID- 18414016 TI - Genetic diversity among Korean bermudagrass (Cynodon spp.) ecotypes characterized by morphological, cytological and molecular approaches. AB - The genus Cynodon comprises ten species. The objective of this study was to evaluate the genetic diversity of Korean bermudagrasses at the morphological, cytological and molecular levels. Morphological parameters, the nuclear DNA content and ploidy levels were observed in 43 bermudagrass ecotypes. AFLP markers were evaluated to define the genetic diversity, and chromosome counts were made to confirm the inferred cytotypes. Nuclear DNA contents were in the ranges 1.42 1.56, 1.94-2.19, 2.54, and 2.77-2.85 pg/2C for the triploid, tetraploid, pentaploid, and hexaploid accessions, respectively. The inferred cytotypes were triploid (2n = 3x = 27), tetraploid (2n = 4x = 36), pentaploid (2n = 5x = 45), and hexaploid (2n = 6x = 54), but the majority of the collections were tetraploid (81%). Mitotic chromosome counts verified the corresponding ploidy levels. The fast growing fine-textured ecotypes had lower ploidy levels, while the pentaploids and hexaploids were coarse types. The genetic similarity ranged from 0.42 to 0.94 with an average of 0.64. UPGMA cluster analysis and principle coordinate analysis separated the ecotypes into 6 distinct groups. The genetic similarity suggests natural hybridization between the different cytotypes, which could be useful resources for future breeding and genetic studies. PMID- 18414017 TI - Chloroplastic NAD(P)H dehydrogenase complex and cyclic electron transport around photosystem I. AB - Recent molecular genetics studies have revealed that cyclic electron transport around photosystem I is essential for normal photosynthesis and growth of plants. Chloroplastic NAD(P)H dehydorgenase (NDH) complex, a homologue of the complex I in respiratory electron transport, is involved in one of two cyclic pathways. Recent studies on the function and structure of the NDH complex are reviewed. PMID- 18414018 TI - Dopamine receptor interacting proteins (DRIPs) of dopamine D1-like receptors in the central nervous system. AB - Dopamine is a major neurotransmitter in the mammalian central nervous system (CNS) that regulates neuroendocrine functions, locomotor activity, cognition and emotion. The dopamine system has been extensively studied because dysfunction of this system is linked to various pathological conditions including Parkinson's disease, schizophrenia, Tourette's syndrome, and drug addiction. Accordingly, intense efforts to delineate the full complement of signaling pathways mediated by individual receptor subtypes have been pursued. Dopamine D1-like receptors are of particular interest because they are the most abundant dopamine receptors in CNS. Recent work suggests that dopamine signaling could be regulated via dopamine receptor interacting proteins (DRIPs). Unraveling these DRIPs involved in the dopamine system may provide a better understanding of the mechanisms underlying CNS disorders related to dopamine system dysfunction and may help identify novel therapeutic targets. PMID- 18414020 TI - Why cells respond differently to DNA damage: a chromatin perspective. AB - In response to DNA double-stranded breaks (DSBs) cells activate a signaling cascade known as the DNA damage response (DDR) whose main function is to promote the repair of the lesions while it delays cell cycle progression until repair is completed. Whereas most cells respond alike to an equivalent dose of DNA damage, certain degree of heterogeneity exists in the strength of the DDR that is assembled in each individual cell. This variability might be accounted for by erratic changes that aggregate into the inherent noise of biological systems. However, we have recently found that the overall degree of chromatin compaction impinges a direct constrain on the activation of the DDR, providing a simple chromatin-based model to explain the cell-to-cell variability observed in cell populations. We here provide an overview of the available data, including our own, that would be supportive of such a model and discuss how this perspective might be used to explain previous observations. PMID- 18414021 TI - Interplay between microRNAs and RNA-binding proteins determines developmental processes. AB - MicroRNAs (miRNAs) are genes involved in normal development and cancer. They inhibit gene expression by associating with 3'-Untranslated regions (3'UTRs) of messenger RNAs (mRNAs), and are thought to regulate a large proportion of protein coding genes. However, it is becoming apparent that miRNA activity is not necessarily always determined by its expression in the cell. MiRNA activity can be affected by RNA-binding proteins (RBPs). For example, the RNA-binding protein HuR associates with the 3'UTR of the CAT1 mRNA after stress, counteracting the effect of miR-122. Second, we found that the expression of an RNA-binding protein called Dead end (Dnd1) prohibits the function of several miRNAs by blocking the accessibility of target mRNAs.(2) Dnd1 function is essential for proper development of primordial germ cells (PGCs) in zebrafish and mammals, indicating a crucial role for RBP/miRNA interplay on 3'UTRs of mRNAs in developmental decisions. In this perspective we discuss the interplay between RBPs and miRNAs in the context of germ cells and review current observations implicating RBPs in miRNA function. PMID- 18414022 TI - HSP90: the Rosetta stone for cellular protein dynamics? AB - The Hsp90 proteomic network is expansive and includes a variety of cell processes operating within the cytoplasm and nucleoplasm. Though the functional significance of the extensive interactions has not been defined, we suggest that the Hsp90 molecular chaperone machinery promotes dynamic behaviors for client proteins that is critical to achieve homeostasis. A general rapid action by cell factors would permit both proper assembly of biological complexes and efficient transitions between distinct structures. Here, we describe why the properties that are inherent to molecular chaperones place these proteins in a unique position to drive the dynamic cellular environment. PMID- 18414023 TI - Caspase-independent mitotic death (CIMD). AB - The spindle checkpoint, which monitors kinetochore-microtubule attachment, is required for high fidelity of chromosome transmission. A failure in this mechanism causes aneuploidy, thereby promoting progression to tumorigenesis. However, the cell death mechanism that prevents the aneuploidy caused by failure of the spindle checkpoint is yet unknown. We have recently identified a novel type of mitotic cell death, which we term caspase-independent mitotic death (CIMD). In BUB1-deficient (but not MAD2-deficient) cells, CIMD is induced by conditions that activate the spindle checkpoint (i.e., cold shock or treatment with nocodazole, paclitaxel or 17-AAG [17-allylaminogeldanamycin]). CIMD depends on p73, a homolog of p53, but not on p53. It also depends on the apoptosis inducing factor (AIF) and endonuclease G (Endo G), which are effectors of caspase independent cell death. When BUB1 is completely depleted, aneuploidy occurs instead of CIMD. We propose that CIMD can be the cell death mechanism that protects cells from aneuploidy by inducing the death of cells prone to substantial chromosome missegregation. Our study also shows that previous evaluations of the spindle checkpoint activity in mutant or cancer cells by monitoring mitotic index could be misleading. PMID- 18414024 TI - A spoke in the wheel: calcium spikes disrupt yeast cell cycle. AB - A prerequisite of calcium signaling is the maintenance of cytoplasmic free Ca(2+) within a narrow, sub-micromolar range, compatible with cell viability. A variety of stress inducing agents and drugs can lead to a disruption of calcium homeostasis with ensuing toxicity. Amiodarone, an anti-arrhythmic drug with broad range microbicidal effect, has been shown to trigger a calcium burst in the yeast Saccharomyces cerevisiae. Recently, we reported that amiodarone elicits a starvation response, blocks cell cycle progression at multiple stages and represses the transcription of cell cycle genes. Here, we examine the connections between Ca(2+)-calcineurin signaling, nutrient sensing and regulation of the cell cycle. PMID- 18414025 TI - Anchorage-independent cytokinesis as part of oncogenic transformation? AB - Cell anchorage to the extracellular matrix (ECM) controls the cell proliferation in all multicellular organisms and the abrogation of this control is an indicator of cellular transformation. In fact, two distinct periods of the cell cycle are subject to anchorage-dependent regulation. Firstly, anchorage exerts an extensive control of the G(1)-phase, a control that we found to be more rigorous than for example the control by growth factors. Secondly, anchorage regulates the progression through cytokinesis. In order to achieve anchorage-independent growth a cell must circumvent these controls. To this end, we recently found that oncogenic H-RasV12 can provide sufficient signals to overcome the anchorage dependence for cytokinesis. Together with earlier findings on G(1)-phase control, this demonstrates that oncogenic signaling contributes to de-regulation of anchorage-dependence during both the G(1)-phase and the cytokinesis. This also suggests that de-regulated cytokinesis may be part of oncogenic transformation. PMID- 18414026 TI - Silent no more: expression of RNA from telomeres may regulate telomere length. PMID- 18414027 TI - The cytoprotective Nrf2 transcription factor controls insulin receptor signaling in the regenerating liver. AB - The Nrf2 transcription factor is a crucial regulator of the cellular redox homeostasis through its capacity to induce the expression of enzymes, which detoxify reactive oxygen species, and of other antioxidant proteins. Therefore, it plays an important role in the protection from carcinogenesis induced by various insults. In addition, recent results identified a novel role of Nrf2 in tissue repair. In the liver, regeneration after partial hepatectomy was strongly delayed in the absence of Nrf2. This defect was shown to result from transient resistance to insulin and insulin-like growth factor 1 that was caused by chronic oxidative stress in hepatocytes. These results demonstrate a link between Nrf2 deficiency, oxidative stress and insulin resistance, and suggest that activation of this transcription factor could be a novel strategy to improve liver regeneration in patients with acute or chronic liver injury. In addition, it may help to alleviate oxidative stress-induced insulin resistance in the liver and potentially also in other organs. PMID- 18414028 TI - Switching cyclin D-Cdk4 kinase activity on and off. AB - The cyclin-cdks are master regulators of cell proliferation. These serine/threonine kinases are the motors that both start and stop the cell cycle in response to proliferative or antiproliferative signals. They phosphorylate substrates required to trigger orderly cell cycle progression, and thus their activity is tightly regulated in order to prevent inappropriate activation. One of the main interfaces between the extraceullar environment and the cell cycle machinery is the interaction of the cyclin-cdks with two families of stoichiometric cyclin kinase inhibitors (CKIs), the Ink4s and the Cip/Kips. As their name suggests, the CKIs have historically been considered negative regulators of the cyclin-cdks, responsible for rapidly and effectively turning off cyclin-cdk activity. However, the interaction of cyclin D-cdk4 with the Cip/Kip family, and with p27Kip1 in particular, appeared complex. In addition to its ability to inhibit cyclin D-cdk4, p27 appeared to be a required assembly factor for the complex, binding in a non-inhibitory mode at least some of the time. Whether p27 was a cyclin D-cdk4/6 inhibitor or not was controversial, and how it might switch between these two modes was unknown. Arguing for a two state mechanism, we have recently shown that p27 can be both a cdk4 bound-inhibitor and a bound-non-inhibitor, depending on the growth state of the cell. This perspective highlights the significance of this finding in terms of normal cell cycle progression and tumor development. PMID- 18414029 TI - Cholera vaccines for the developing world. AB - Cholera remains as a global public health threat affecting most of the developing world. In endemic areas, young children are most affected. Outbreaks are reported increasingly from more countries. Improvements in water and sanitation may be the mainstays of cholera prevention but in the short term, vaccines provide an alternative in cholera control. Since 1999, the World Health Organization has advocated the use of oral cholera vaccines as an adjunct in the control of cholera. Although internationally available, oral cholera vaccines are not extensively used in public health settings in developing countries where they are most needed. We review oral cholera vaccines that are currently available and in the pipeline including the need for policies that will ensure more extensive usage of these vaccines. PMID- 18414030 TI - Molecular framework underlying pluripotency. AB - Pluripotency is governed by the intricate interplay between genetic and epigenetic factors. Previous studies implicate Oct4, Sox2 and Nanog as core regulators of the transcriptional circuitry in pluripotent cells. The three transcriptional factors and their downstream target genes coordinately promote self-renewal and pluripotency. Significant inroads have also been made into elucidating the epigenetic mechanisms governing the exit from a self-renewal state. The recent success in reprogramming both mouse and human fibroblast cells into a pluripotent state has marked a new era in stem cell research and is pertinent to a comprehensive understanding of pluripotency. Reprogramming can be initiated by the introduction of few defined factors. However, the molecular mechanisms driving reprogramming remain largely obscure and await further investigations. Here, we explore the molecular framework governing pluripotency in the context of pluripotent ES cells and the reprogrammed iPS cells. PMID- 18414031 TI - Break-induced replication: what is it and what is it for? AB - Homologous recombination (HR) is considered to be an error-free mechanism for the repair of DNA double-strand breaks (DSBs). Indeed, most DSB repair events occur by a non-crossover mechanism limiting loss of heterozygosity (LOH) for markers downstream of the site of repair and preventing chromosome rearrangements. However, DSBs that arise by replication fork collapse or by erosion of uncapped telomeres have only one free end and are thought to repair by strand invasion into a homologous duplex DNA followed by replication to the chromosome end (break induced replication, BIR). As BIR from one of the two ends of a DSB would result in a long tract of LOH it suggests BIR is suppressed when DSBs have two ends in order for repair to occur by a more conservative HR mechanism. Recent studies showed that BIR can occur by several rounds of strand invasion, DNA synthesis and dissociation resulting in chromosome rearrangements when dissociation and reinvasion occur within dispersed repeated sequences. Thus template switching BIR can be highly mutagenic and this process could be important for genome evolution and disease development. PMID- 18414033 TI - Downregulation of c-jun results in apoptosis-mediated anti-osteosarcoma activity in an orthotopic model. AB - c-jun has been found to be upregulated in a variety of cancers including osteosarcoma. DNAzymes are oligonucleotides capable of specific downregulation of target genes. c-jun knockdown-mediated apoptosis in osteosarcoma cells involved caspases-1, -2 and -8, but not the Fas/FasL pathway. A c-jun DNAzyme, encapsulated within a novel cationic multilamellar vesicle liposome, inhibited the growth and metastasis of osteosarcoma in an orthotopic spontaneously metastasising model of the disease. The 60 nm DDAB:DOPE liposome was formulated using ethanol injection/extrusion. Clinically, downregulation of c-jun may proffer an improved treatment outcome for these tumours originating in bone. PMID- 18414032 TI - Unique and important consequences of RECQ1 deficiency in mammalian cells. AB - Five members of the RecQ subfamily of DEx-H-containing DNA helicases have been identified in both human and mouse, and mutations in BLM, WRN, and RECQ4 are associated with human diseases of premature aging, cancer, and chromosomal instability. Although a genetic disease has not been linked to RECQ1 mutations, RECQ1 helicase is the most highly expressed of the human RecQ helicases, suggesting an important role in cellular DNA metabolism. Recent advances have elucidated a unique role of RECQ1 to suppress genomic instability. Embryonic fibroblasts from RECQ1-deficient mice displayed aneuploidy, chromosomal instability, and increased load of DNA damage.(1) Acute depletion of human RECQ1 renders cells sensitive to DNA damage and results in spontaneous gamma-H2AX foci and elevated sister chromatid exchanges, indicating aberrant repair of DNA breaks.(2) Consistent with a role in DNA repair, RECQ1 relocalizes to irradiation induced nuclear foci and associates with chromatin.(2) RECQ1 catalytic activities(3) and interactions with DNA repair proteins(2,4,5) are likely to be important for its molecular functions in genome homeostasis. Collectively, these studies provide the first evidence for an important role of RECQ1 to confer chromosomal stability that is unique from that of other RecQ helicases and suggest its potential involvement in tumorigenesis. PMID- 18414034 TI - Guilt by association: PAX3-FOXO1 regulates gene expression through selective destabilization of the EGR1 transcription factor. AB - Human cancer cells frequently harbor chromosomal translocations that create chimeric fusion genes. The t(2;13) translocation is characteristic of the pediatric muscle tumor, alveolar rhabdomyosarcoma, and produces the chimeric transcription factor, PAX3-FOXO1, that contains the DNA binding elements of PAX3 and the transcriptional activation domain of FOXO1. Experiments designed to determine how PAX3-FOXO1 expression contributes to the development of muscle cell derived tumors resulted in the discovery that the fusion protein misregulates gene expression and interrupts myogenic differentiation through a unique gain of function mechanism. These results yield new insight into how tumor-associated genetic alterations increase the likelihood of cancer formation and may lead to new therapeutic approaches. PMID- 18414035 TI - Untangling the unfolded protein response. AB - Resistance to current cancer therapies has forced scientists to investigate new avenues of therapy distinct from those aimed at single targets, to strategies based on targeting families of proteins, on which cancers rely for their ability to survive stress. Two such protein families are the heat shock proteins (HSP), especially the HSP90 family, and proteins involved in mediating the unfolded protein response (UPR). HSP90 stabilises key survival factors in cancer cells including AKT, ERB2 and HIF1alpha, which alone makes HSP90 inhibitors extremely interesting as potential therapies. In addition targeting HSP90 can destabilise the UPR inducing cell death. A broad range of cancer-types rely on the UPR to correctly fold key signalling proteins properly, as well as to allow the cell to cope with the hypoxic environment associated with tumour development. These associations suggest that a range of tumours may be targeted using HSP90 inhibitors and that the development of specific inhibitors of the UPR may be of interest. In this article, based on work in multiple myeloma, we highlight the importance of targeting multiple signalling pathways simultaneously, using the UPR and heat shock proteins as examples, as a means of effectively killing cancer cells. PMID- 18414036 TI - IAPs: more than just inhibitors of apoptosis proteins. AB - Inhibitors of apoptosis proteins (IAPs) are a conserved family of proteins identified in species ranging from virus, yeasts, nematodes, fishes, flies and mammals. The common structural feature is the presence of at least one Baculovirus IAP Repeat (BIR) domain. Hence, IAPs are also known as BIR-containing proteins (BIRCs). Most of them display anti-apoptotic properties when overexpressed. In drosophila, IAPs are sufficient and necessary to promote cell survival through a direct regulation of apoptotic proteases called caspases. In mammals, BIRC4/XIAP, the most studied IAP member can directly inhibit the activity of caspase-3, 7 and 9. However, this activity is not conserved in other IAPs and physiological relevancies of such anti-caspase activities are still discussed. A detailed analysis of IAP-deficient mice or derived cells, deletion experiments performed in drosophila and zebrafish, or research of protein partners have revealed the importance of IAPs in adaptive response to cellular stress, in cell proliferation, differentiation, signaling, motility and in immune response. This review discusses recent data that help understanding of cellular functions of IAPs. PMID- 18414039 TI - The tumor suppressor p53: cancer and aging. AB - Aging, like many other biological processes, is subject to regulation by genes that reside in pathways that have been conserved during evolution. The insulin/ IGF-1 pathway, mTOR pathway and p53 pathway are among those conserved pathways that impact upon longevity and aging-related diseases such as cancer. Most cancers arise in the last quarter of life span with the frequency increasing exponentially with time, and mutation accumulation in critical genes (e.g., p53) in individual cells over a lifetime is thought to be the reason. Recently, we found that the efficiency of the p53 response to stress declines significantly with age in mice, and the time of onset of this decreased p53 response correlates with the life span of mice. Given the crucial role of the p53 in tumor prevention, this decline in p53 activity at older ages in animals could contribute to the observed dramatic increases in cancer frequency, and provides a plausible explanation for the correlation between tumorigenesis and aging in addition to the accumulation of DNA mutations over lifetime. We discuss here the coordination and communication between the p53 pathway and the IGF-1-mTOR pathways, and their possible impact on cancer and longevity. PMID- 18414037 TI - The role of the PTEN/AKT Pathway in NOTCH1-induced leukemia. AB - Activating mutations in NOTCH1 are the most prominent genetic abnormality in T cell acute Lymphoblastic Leukemia (T-ALL) and inhibition of NOTCH1 signaling with gamma-secretase inhibitors (GSIs) has been proposed as targeted therapy in this disease. However, most T-ALL cell lines with mutations in NOTCH1 fail to respond to GSI therapy. Using gene expression profiling and mutation analysis we showed that mutational loss of PTEN is a common event in T-ALL and is associated with resistance to NOTCH inhibition. Furthermore, our studies revealed that NOTCH1 induces upregulation of the PI3K-AKT pathway via HES1, which negatively controls the expression of PTEN. This regulatory circuitry is evolutionary conserved from Drosophila to humans as demonstrated by the interaction of overexpression of Delta and Akt in a model of Notch-induced transformation in the fly eye. Loss of PTEN and constitutive activation of AKT in T-ALL induce increased glucose metabolism and bypass the requirement of NOTCH1 signaling to sustain cell growth. Importantly, PTEN-null/GSI resistant T-ALL cells switch their oncogene addiction from NOTCH1 to AKT and are highly sensitive to AKT inhibitors. These results should facilitate the development of molecular therapies targeting NOTCH1 and AKT for the treatment of T-ALL. PMID- 18414040 TI - The stem cells of the neural crest. AB - In the vertebrate embryo, the neurectodermal neural crest cells (NCC) have remarkably broad potencies, giving rise, after a migratory phase, to neurons and glial cells in the peripheral nervous system, and to skin melanocytes, being all designated here as "neural" derivatives. NC-derived cells also include non neural, "mesenchymal" cell types like chondrocytes and bone cells, myofibroblasts and adipocytes, which largely contribute to the head structures in amniotes. Similar to the blood cell system, the NC is therefore a valuable model to investigate the mechanisms of cell lineage diversification in vertebrates. Whether NCC are endowed with multiple differentiation potentials or if, conversely, they are a mosaic of different committed cells is an important ongoing issue to understand the ontogeny of NC derivatives in normal development and pathological conditions. Here we focus on recent findings that established the presence in the early migratory NC of the avian embryo, of a multipotent progenitor endowed with both mesenchymal and neural differentiation capacities. This "mesenchymal-neural" clonogenic cell lies upstream of all the other NC progenitors known so far and shows increased frequency when single cell cultures are treated with the Sonic Hedgehog signaling molecule. These findings are discussed in the context of the broad potentials of NC stem cells recently evidenced in certain adult mammalian tissues. PMID- 18414041 TI - Chk1 is activated at the midblastula transition in Xenopus laevis embryos independently of DNA content and the cyclin E/Cdk2 developmental timer. AB - Cell cycle checkpoints that are engaged in response to damaged and unreplicated DNA may serve additional, constitutive functions. In the developing Xenopus laevis embryo, the checkpoint kinase Chk1 is transiently activated at the midblastula transition (MBT), a period of extensive cell cycle remodeling including the acquisition of cell cycle checkpoints. The timing of many cell cycle remodeling events at the MBT, such as the lengthening of cell cycles, depends upon a critical nucleocytoplasmic (N/C) ratio. However, other events, including the degradation of maternal cyclin E, do not depend upon the N/C ratio, and are regulated by an autonomous developmental timer. To better understand what regulates Chk1 activation at the MBT, embryos were treated with aphidicolin, at different developmental times and for different lengths of time, to reduce the DNA content at the MBT. Chk1 was activated at the MBT in these embryos establishing that Chk1 activation occurs independently of the N/C ratio. Cdc25A is normally phosphorylated by Chk1 at the MBT and then degraded. The degradation of Cdc25A demonstrated partial dependence on DNA content, suggesting that factors other than Chk1 regulate its degradation. When the cyclin E developmental timer was disrupted with the Cdk2 inhibitor delta34-Xic1, Chk1 was still activated at the MBT, indicating that activation of Chk1 at the MBT was not directly linked to the cyclin E timer. Conversely, unreplicated or damaged DNA, delayed the degradation of cyclin E at the MBT, indicating that the cyclin E/Cdk2 timer is sensitive to engagement of cell cycle checkpoints. PMID- 18414042 TI - Both SCF(Cdc4alpha) and SCF(Cdc4gamma) are required for cyclin E turnover in cell lines that do not overexpress cyclin E. AB - The ubiquitin-mediated turnover of cyclin E is regulated by phosphorylation and the activity of the ubiquitin ligase SCF(Cdc4) (also known as SCF(Fbw7)). In 293A cells, SCF complexes containing two different Cdc4 isoforms, alpha and gamma, are required for efficient cyclin E ubiquitylation. Whereas SCF(Cdc4gamma) ubiquitylates cyclin E directly, SCF(Cdc4alpha) serves as a cofactor for Pin1 mediated prolyl isomerization of the cyclin E phosphodegron, essential to potentiate ubiquitylation. In the current study, we show that the requirement for both Cdc4alpha and gamma is general, except in cell lines where cyclin E is expressed at an elevated level. Under these circumstances, Cdc4alpha is sufficient for cyclin E turnover. Furthermore, the requirement for Cdc4gamma can be bypassed by ectopic overexpression of cyclin E. PMID- 18414043 TI - Adaptive autophagy in Alexander disease-affected astrocytes. AB - The ubiquitin-proteasome and autophagy-lysosomal pathways are the two main routes of protein and organelle clearance in eukaryotic cells. The proteasome system is responsible for unfolded, short-lived proteins, which precludes the clearance of oligomeric and aggregated proteins, whereas macroautophagy, a process generally referred to as autophagy, mediates mainly the bulk degradation of long-lived cytoplasmic proteins, large protein complexes or organelles.(1) Recently, the autophagy-lysosomal pathway has been implicated in neurodegenerative disorders as an important pathway for the clearance of abnormally accumulated intracellular proteins, such as huntingtin, tau and mutant and modified alpha-synuclein.(1-6) Our recent study illustrated the induction of adaptive autophagy in response to mutant glial fibrillary acidic protein (GFAP) accumulation in astrocytes, in the brains of patients with Alexander disease (AxD), and in mutant GFAP knock-in mouse brains.(7) This autophagic response is negatively regulated by mammalian target of rapamycin (mTOR). The activation of p38 MAPK by GFAP accumulation is responsible for mTOR inactivation and the induction of autophagy. We also found that the accumulation of GFAP impairs proteasome activity.(8) In this commentary we discuss the potential compensatory relationship between an impaired proteasome and activated autophagy, and propose that the MLK-MAPK (mixed lineage kinase mitogen-activated protein kinase) cascade is a regulator of this crosstalk. PMID- 18414044 TI - The oncogene c-Myc coordinates regulation of metabolic networks to enable rapid cell cycle entry. AB - The c-myc proto-oncogene is rapidly activated by serum and regulates genes involved in metabolism and cell cycle progression. This gene is thereby uniquely poised to coordinate both the metabolic and cell cycle regulatory events required for cell cycle entry. However, this function of Myc has not been evaluated. Using a rat fibroblast model of isogenic cell lines, myc(-/-), myc(+/-), myc(+/+) and myc(-/-) cells with an inducible c-myc transgene (mycER), we show that the Myc protein programs cells to utilize both oxidative phosphorylation and glycolysis to drive cell cycle progression. We demonstrate this coordinate regulation of metabolic networks is essential, as specific inhibitors of these pathways block Myc-induced proliferation. Metabolic events temporally correlated with cell cycle entry include increased oxygen consumption, mitochondrial function, pyruvate and lactate production, and ATP generation. Treatment of normal cells with inhibitors of oxidative phosphorylation recapitulates the myc(-/-) phenotype, resulting in impaired cell cycle entry and reduced metabolism. Combined with a kinetic expression profiling analysis of genes linked to mitochondrial function, our study indicates that Myc's ability to coordinately regulate the mitochondrial metabolic network transcriptome is required for rapid cell cycle entry. This function of Myc may underlie the pervasive presence of Myc in many human cancers. PMID- 18414045 TI - RB status governs differential sensitivity to cytotoxic and molecularly-targeted therapeutic agents. AB - The retinoblastoma tumor suppressor (RB) is frequently inactivated in human cancers and has been shown to modulate the anti-proliferative effects of DNA damaging therapies. However, the impact of RB loss on response to disparately functioning cytotoxic agents as well as targeted therapies is poorly understood. Here 3T3-immortalized and Ras-transformed mouse adult fibroblasts (MAFs) containing conditional RB alleles were utilized to investigate the consequence of RB loss on cellular response to cytotoxic agents and therapies targeting the MEK and PI3K pathways. Using these models, we demonstrate that RB deficiency is associated with bypass of therapy-induced checkpoints in response to both cytotoxic and targeted treatments. Interestingly, while checkpoint bypass following treatment with cytotoxic therapy results in an agent specific increase in drug sensitivity, checkpoint bypass following treatment targeting MEK and PI3K function results in increased cellular proliferation. These results indicate that RB status differentially impacts therapeutic response and should be considered when evaluating the efficacy of molecularly targeted therapeutics. PMID- 18414046 TI - A Hippo in the ointment: MST signalling beyond the fly. AB - The regulation of cell cycle and apoptosis is fundamental to the control of cell growth and organism homeostasis. Failure to efficiently regulate these processes often results in the increased cell growth observed in tumours. Accumulation of genetic lesions frequently eliminates these regulatory steps so it is imperative that multiple signalling pathways are employed to ensure that efficient control is maintained. Over the last few years a novel signalling pathway entered the limelight that prevents inappropriate activation of the cell cycle and can elicit apoptosis to limit cell numbers. Denoted the MST/hippo pathway, it is involved in regulating cell number in organism development and tumour progression. Here we aim to review the evidence for a conserved pathway from flies to mammals, and of equal importance to initiate the discussion on the additional cellular and signalling processes that have been adopted by this pathway to achieve further regulation and diversified cellular outcomes in mammals. PMID- 18414047 TI - p53: a new player in reproduction. AB - The roles of the p53 protein in tumor suppression have been firmly established. However, the functions of this protein under normal conditions or in the absence of stress, if any, have remained a mystery. In humans, some alleles containing a functional single nucleotide polymorphism in the p53 gene and its negative regulator, the Mdm2 gene, are under positive selection over evolutionary time frames, suggesting that the p53 pathway might have important functions that are optimized and selected for by evolutionary or reproductive pressures. Indeed, a recent study demonstrated a new function for the p53 protein in the regulation of maternal reproduction in mice, through transcriptional regulation of leukemia inhibitory factor (LIF), a novel p53 target gene. Sufficient uterine LIF levels are essential for the implantation of blastocysts or early embryos into the uterus. p53 deficient (p53(-/-)) female mice have a reduced pregnancy rate and litter size, due to impaired implantation resulting from decreased uterine LIF levels. Administration of LIF to pregnant p53(-/-) mice restored maternal reproduction by improving implantation. An association has been reported between women carrying the p53 codon 72 polymorphism (a proline to arginine change) with recurrent implantation failure, suggesting a similar function for p53 in humans. These findings of a new function for the p53 protein in reproduction may help to explain the observed evolutionary selection of some alleles of the p53 and Mdm2 genes. This may also be an excellent example of antagonistic pleiotrophy. PMID- 18414048 TI - Cell-surface MMP-9 regulates the invasive capacity of leukemia blast cells with monocytic features. AB - The metalloprotease 9 (MMP-9), a known mediator of tumour invasion, is secreted as a 92 kDa pro-form but a non-secreted variant of 85 Kda has been described. The importance of this variant pro-form in tumor progression remains poorly defined. We previously showed that the DNA repair protein Ku interacts at the cell surface of leukaemia cell lines with the 85 Kda pro-form of MMP-9 and these Ku/MMP-9 complexes regulates cell invasion, highlighting their importance in haematological malignancies. We demonstrate here that all samples of acute myeloid leukaemia (AML) blasts purified from bone marrow of 16 affected patients express the 85 Kda form of MMP-9. However, only AML that display monocytic lineage markers (AML4/5) express this form at the cell surface with co-expression of the membrane associated form of Ku. Blocking antibodies directed against Ku or MMP-9 specifically inhibited cell invasion of those expressing Ku/MMP-9 on the cell surface. The membrane form of Ku might represent an important factor in the exposition to the cell surface of this specific MMP-9 pro-form in AML with monocytic features. These results might have important functional significance in the occurrence of extra-medullar infiltrates of leukaemia cells that occurs frequently during the onset of monocyte-related AML sub-types. PMID- 18414049 TI - The pre-clinical development of MDM2 inhibitors in chronic lymphocytic leukemia uncovers a central role for p53 status in sensitivity to MDM2 inhibitor-mediated apoptosis. AB - Inhibitors of the MDM2-p53 interaction are actively being developed as anti cancer agents. Drug-induced interference with the MDM2 E3 ligase function or with MDM2 protein-protein interactions abrogates tonic suppression and destruction of the p53 protein; consequently, p53 steady state levels rise resulting in the induction of p53-dependent anti-proliferative and pro-apoptotic genes. Some cancerous cells harboring wild type p53 respond to MDM2 inhibitor-induced elevated p53 protein levels with apoptotic cell death while non-malignant cells, for poorly understood reasons, appear relatively resistant. Deciphering the mechanisms of resistance or susceptibility to MDM2 inhibitor-induced cancer cell death is of significant importance for the clinical development and applications of MDM2 inhibitory compounds and serves to illuminate aspects of MDM2 and p53 biology. Using data from ex vivo MDM2 inhibitor treatment of a large cohort of molecularly highly characterized CLL cases, we were able to demonstrate the central role of p53 status as a determinant of resistance in this common leukemia. In the context of these experimental findings, we summarize pertinent knowledge of the biology of p53, MDM2, p53 target genes and MDM2 binding proteins. Finally, using data from a large SNP-array-based high-density genomic profiling study in CLL, we summarize the genomic copy number and allele status for important p53 effector genes as well as for MDM2 binding/target proteins, thus demonstrating the power of high resolution genomic analysis in support of targeted drug development. PMID- 18414050 TI - MiRSNPs or MiR-polymorphisms, new players in microRNA mediated regulation of the cell: Introducing microRNA pharmacogenomics. AB - MicroRNAs are evolutionarily conserved small non-coding RNAs known to inhibit the translation of proteins by binding to the target transcript in the 3' untranslated region. Functional polymorphisms in 3' UTRs of several genes have been reported to be associated with diseases by affecting gene expression. The mechanism by which these polymorphisms affect gene expression and induce variability in a cell is not well understood. It has been suggested that these polymorphisms may interfere with regulatory elements that bind to untranslated region of a gene. Recently, a novel class of functional polymorphisms termed miRSNPs/polymorphisms was reported. defined as a polymorphism present at or near a microRNA binding sites of functional genes that can affect gene expression by interfering with a miRNA function. The work elucidated the mechanism of a functional miRSNP 829C-->T present in 3' UTR of dihydrofolate reductase, an important drug target. The SNP interferes with the miR24 microRNA function and leads to DHFR over expression and methotrexate resistance. In this article we highlight the importance of these miRSNPs or miR-polymorphisms in gene regulation and the mechanism by which these miRSNPs can induce variability in the SNP expressing mutant cell by using drug resistance as an example. PMID- 18414051 TI - The level of oncogene H-Ras correlates with tumorigenicity and malignancy. AB - Normal bovine adrenocortical cells and some human fibroblasts can be transformed by SV40T and H-Ras in a Ras-dependent manner. We recently reported that high levels of Ras derived from 5' LTR of retrovirrus can induce highly malignant and fast growing tumors, while lower levels of Ras derived from internal ribosome entry site (IRES) promotes slower tumor growth and loss of malignancy. Ras derived from CMV promoters resulted in much lower Ras levels and loss of tumor malignancy and growth. Further studies showed that the tumors formed in the presence of lower levels of Ras and dominant negative P53 (P53DD) had fewer apoptotic cells and grew faster than the tumors formed from cells with same level Ras and SV40T. Our studies suggest that low levels of Ras are insufficient to inhibit apoptosis induced by pRb inactivation. In contrast, high levels of Ras not only allow normal cells to exit senescence and form tumors, but also protect against pRb inhibition-induced cell apoptosis. PMID- 18414053 TI - Resveratrol: a multitargeted agent for age-associated chronic diseases. AB - Extensive research within the last decade has revealed that most chronic illnesses such as cancer, cardiovascular and pulmonary diseases, neurological diseases, diabetes, and autoimmune diseases exhibit dysregulation of multiple cell signaling pathways that have been linked to inflammation. Thus mono-targeted therapies developed for the last two decades for these diseases have proven to be unsafe, ineffective and expensive. Although fruits and vegetables are regarded to have therapeutic potential against chronic illnesses, neither their active component nor the mechanism of action is well understood. Resveratrol (trans-3, 5, 4'-trihydroxystilbene), a component of grapes, berries, peanuts and other traditional medicines, is one such polyphenol that has been shown to mediate its effects through modulation of many different pathways. This stilbene has been shown to bind to numerous cell-signaling molecules such as multi drug resistance protein, topoisomerase II, aromatase, DNA polymerase, estrogen receptors, tubulin and F1-ATPase. Resveratrol has also been shown to activate various transcription factor (e.g; NFkappaB, STAT3, HIF-1alpha, beta-catenin and PPAR-gamma), suppress the expression of antiapoptotic gene products (e.g; Bcl-2, Bcl-X(L), XIAP and survivin), inhibit protein kinases (e.g; src, PI3K, JNK, and AKT), induce antioxidant enzymes (e,g; catalase, superoxide dismutase and hemoxygenase-1), suppress the expression of inflammatory biomarkers (e.g., TNF, COX-2, iNOS, and CRP), inhibit the expression of angiogenic and metastatic gene products (e.g., MMPs, VEGF, cathepsin D, and ICAM-1), and modulate cell cycle regulatory genes (e.g., p53, Rb, PTEN, cyclins and CDKs). Numerous animal studies have demonstrated that this polyphenol holds promise against numerous age-associated diseases including cancer, diabetes, Alzheimer, cardiovascular and pulmonary diseases. In view of these studies, resveratrol's prospects for use in the clinics are rapidly accelerating. Efforts are also underway to improve its activity in vivo through structural modification and reformulation. Our review describes various targets of resveratrol and their therapeutic potential. PMID- 18414054 TI - Stress-dependent changes in the properties of p53 complexes by the alternative translation product p53/47. AB - P53 plays a key role in the cellular response to damage exposure and in preventing the development of human cancers. Activation of p53 results in changes in the expression of a large number of gene products. However, relatively little is still known how p53 activation differentiates between different types of damages in different types of tissues or how this triggers either an apoptotic response or cell cycle arrest and DNA repair. The p53 message is translated into two products with distinct activities and stabilities through alternative mechanisms of initiation. P53/47 is initiated 40 codons down stream of the full length p53 and does not include the binding site for the E3 ubiquitin ligase Mdm2 or the transactivation domain I but retains the capacity to form p53 hetero- and homo-oligomers. Here we report that p53/47 controls the folding, the oligomerisation and the post-translational modification of p53 complexes and that it diversifies p53 properties in a cell stress-dependent fashion. P21 expression, for example, is under normal conditions not affected by p53/47 but is induced 18 fold after treatment of cells with the DNA damaging drug doxorubicin. This is accompanied by the recruitment of p53/47 to the p21 promoter. PMID- 18414052 TI - Role of the mammalian SWI/SNF chromatin remodeling complex in the cellular response to UV damage. AB - Mammalian cells exhibit complex cellular responses to DNA damage, including cell cycle arrest, DNA repair and apoptosis. Defects in any one of these responses can result in carcinogenesis. Absence of the chromatin remodeling complex Swi/Snf is found in many instances of cancer, and we have investigated its role in the UV damage response. The human carcinoma cell line SW13 is deficient in Swi/Snf and is very sensitive to UV radiation. In contrast, SW13 cells with ectopic Brg1 expression regain active Swi/Snf and become significantly more resistant to UV radiation. Sensitivity to UV light correlates well with dramatic UV induced apoptosis in SW13 cells, but not in SW13 cells expressing Brg1. We show that SW13 cells synchronized at the G(1)/S border progress into S phase after UV irradiation, and this checkpoint deficiency is corrected after Brg1 expression is restored. Interestingly, Brg1 expression in SW13 cells restores expression of two DNA damage responsive genes, Gadd45a and p21. Furthermore, Gadd45a induction and p21 degradation were observed in the Brg1-expressing SW13 cells after UV irradiation. Our findings demonstrate that Swi/Snf protects cells against deleterious consequences of UV induced DNA damage. These results also indicate that Swi/Snf may modulate checkpoint activation after UV damage via regulation of the two PCNA-binding proteins Gadd45a and p21. PMID- 18414055 TI - Induction of differentiation of human leukemia cells by combinations of COX inhibitors and 1,25-dihydroxyvitamin D3 involves Raf1 but not Erk 1/2 signaling. AB - Differentiation therapy of cancer is being explored as a potential modality for treatment of myeloid leukemia, and derivatives of vitamin D are gaining prominence as agents for this form of therapy. Cyclooxygenase (COX) inhibitors have been reported to enhance 1,25-dihydroxyvitamin D(3) (1,25D)-induced monocytic differentiation of promyeloblastic HL60 cells, but the mechanisms of this effect are not fully elucidated, and whether this potentiation can occur in other types of myeloid leukemia is not known. We found that combination treatment with 1,25D and non-specific COX inhibitors acetyl salicylic acid (ASA) or indomethacin can robustly potentiate differentiation of other types of human leukemia cells, i.e., U937, THP-1, and that ASA +/- 1,25D is effective in primary AML cultures. Increased cell differentiation is paralleled by arrest of the cells in the G(1) phase of the cell cycle, and by increased phosphorylation of Raf1 and p90RSK1 proteins. However, there is no evidence that this increase in phosphorylation of Raf1 is transmitted through the ERK module of the MAPK signaling cascade. Transfection of small interfering (si) RNA to Raf1 decreased differentiation of U937 cells induced by a combination of ASA or indomethacin with 1,25D. However, phosphorylation levels of ERK1/2, though not of p90RSK, were increased when P-Raf1 levels were decreased by the siRNA, suggesting that in this system the ERK module does not function in the conventional manner. Identification of the strong antiproliferative activity of ASA/1,25D combinations associated with monocytic differentiation has implications for cancer chemoprevention in individuals who have a predisposition to myeloid leukemia. PMID- 18414056 TI - Mice lacking both mixed-lineage kinase genes Mlk1 and Mlk2 retain a wild type phenotype. AB - The mitogen-activated protein kinase kinase kinases of the mixed-lineage kinase (MLK) family have been shown to activate the c-Jun N-terminal kinase (JNK) mitogen-activated protein kinase (MAPK) pathway, and to regulate the other two principal MAPK cascades, p38 and extracellular signal-regulated kinase (ERK). Although there is growing evidence for their involvement in neuronal cell death leading to neurodegenerative disorders, little in vivo data is available for the members of this family of kinases. Here, we report that the inactivation of mouse Mlk1 and Mlk2 genes. Mlk1(-/-) and Mlk2(-/-) mice were found to be viable and healthy. Surprisingly, mice carrying the compound Mlk1/Mlk2 null mutations were also found to be viable, fertile and to have a normal life span. The nervous system, testis and kidney, the major sites of MLK1 and 2 expression, all appear normal, as do other organs where these kinases were found to be more weakly expressed. Surprisingly, developmental neuronal programmed cell death, another potential target for MLK family members, was also found to be unaffected. Our results suggest that there is extensive functional redundancy between MLK1/MLK2 and the other member of the family, MLK3, which is also not required for survival in mouse. PMID- 18414057 TI - Synergistic effects of the purine analog sulfinosine and curcumin on the multidrug resistant human non-small cell lung carcinoma cell line (NCI-H460/R). AB - Multidrug resistance (MDR) is the main obstacle to a successful chemotherapy of lung cancer. We tested the potential of sulfinosine and curcumin, alone and in combination, for modulating MDR in the human resistant, non-small cell lung carcinoma cell line (NCI-H460/R). First, we determined the mutational status of the p53 gene in NCI-H460/R cells by PCR-SSCP and DNA sequencing and identified mutations which could at least partially contribute to the development of the MDR phenotype. The effects of sulfinosine and curcumin were studied, both separately and in combination, at the level of cytotoxicity, cell cycle distribution and gene expression. Sulfinosine displayed dose-dependent growth inhibition in both resistant and control sensitive cell lines, whereas curcumin considerably inhibited their growth only at relatively high doses. When sulfinosine was combined with a low dose of curcumin the drugs exerted a synergistic cytotoxic effect in NCI-H460/R cells. The expression of MDR-related genes mdr1, gst-pi and topo IIalpha, was altered by sulfinosine and curcumin. The most pronounced effect was observed when the agents were applied together. Sulfinosine and curcumin caused perturbations in cell cycle distribution in the NCI-H460/R cell line. The combination of the two drugs induced a more pronounced cell cycle arrest in S and G(2)/M in NCI-H460/R cells. Our results show that sulfinosine and curcumin overcome MDR in non-small cell lung carcinoma cell line (NSCLC), especially in combination despite the presence of a mutated p53 gene. PMID- 18414058 TI - Cyclin B1 is rate limiting but not essential for mitotic entry and progression in mammalian somatic cells. AB - Cyclin B1 should have some rate limiting function for cell cycle progression. To test this, we measured the effect of siRNA-mediated depletion of cyclin B1 on mitotic entry and timing. We depleted cyclin B1 in HeLa and hTert-RPE1 cells to levels equivalent or below those achieved in the telophase-to-G(1) window. Average cyclin B1/Cdk1 activity was measured in HeLa cells and depleted by approximately 99%. In both cell lines, this caused approximately 20% increase in the G(2) and approximately 20% increase the M traverse time. However, co depletion of cyclin B1 and B2 induced a profound increase in G(2) cells, a dramatic reduction in mitotic cells, and an increase in a 4C cycling population. We conclude that any residual levels of cyclin B1 were not sufficient to promote stable mitotic entry and transition in absence of normal levels of cyclin B2. Therefore, we conclude that B cyclin is necessary for mitosis but cyclin B1 is not. Nocodazole treated, cyclin B1-depleted HeLa cells arrested but exited that arrest at higher rates than controls, suggesting that the duration of the spindle checkpoint was affected. In B1 depleted cells, population growth was delayed but evidence of cell death was not consistently observed. A strong phenotype of mitotic chromosomal aberration was observed in HeLa cells depleted for either cyclin but not in RPE cells. In B1 or B2 depleted cells, maloriented chromosomes at metaphase were increased 10 fold and one third of affected metaphase cells entered anaphase without congression. Lagging chromosomes at anaphase were dramatically increased. The aggregate evidence from our study and others suggests that the common effect of cyclin B1 depletion is mild cell cycle perturbation. Lack of uniformity in other phenotypes suggest that these are low penetrance effects that are exacerbated or compensated in some systems by other mechanisms. PMID- 18414059 TI - Repression of hedgehog signal transduction in T-lineage cells increases TCR induced activation and proliferation. AB - Hedgehog proteins signal for differentiation, survival and proliferation of the earliest thymocyte progenitors, but their functions at later stages of thymocyte development and in peripheral T-cell function are controversial. Here we show that repression of Hedgehog (Hh) pathway activation in T-lineage cells, by expression of a transgenic repressor form of Gli2 (Gli2DeltaC2), increased T-cell differentiation and activation in response to TCR signalling. Expression of the Gli2DeltaC2 transgene increased differentiation from CD4(+)CD8(+) to single positive thymocyte, and increased peripheral T cell populations. Gli2DeltaC2 T cells were hyper-responsive to activation by ligation of CD3 and CD28: they expressed cell surface activation markers CD69 and CD25 more quickly, and proliferated more than wild-type T-cells. These data show that Hedgehog pathway activation in thymocytes and T-cells negatively regulates TCR-dependent differentiation and proliferation. Thus, as negative regulators of TCR-dependent events, Hh proteins provide an environmental influence on T-cell fate. PMID- 18414060 TI - Requirement of aurora-A kinase in astral microtubule polymerization and spindle microtubule flux. AB - Mitotic Aurora-A kinase was found to be required for formation of bipolar spindle, ensuring accurate chromosome segregation in mitosis. Recently, Aurora-A was shown to promote Ran-GTP-induced spindle formation and astral microtubule development. Here, by selective immunodepletion, we showed that Aurora-A was required for centrosome- but not Ran-GTP-induced astral microtubule formation in Xenopus egg extracts. Aurora-A enhanced microtubule polymerization in both centrosome- and Ran-GTP-induced aster assemblies: shortening the timing of aster assembly and increasing the aster size. Indeed, adding of Aurora-A protein alone induced microtubule clustering, which was abrogated by Aurora kinase inhibitory small molecule ZM447439. In addition, we showed that Aurora-A was indispensable for Ran-GTP-induced bipolar spindle formation. Inhibition of Aurora-A activity by adding of kinase inactive dominant mutant led to spindle collapse and formation of monopolar spindle whereas minus-end motor protein dynein/dynactin inhibitor p50/dynamitin rescued the bipolar structure. Lastly, we revealed that Aurora-A was necessary for microtubule poleward flux and this requirement depended on kinase activity. Thus, we showed that Aurora-A promoted microtubule polymerization and maintained microtubule flux in ensuring proper bipolar spindle assembly. PMID- 18414061 TI - Influenza vaccine for high-risk non-elderly adults: a national survey of subspecialists. AB - Despite long-standing recommendations for non-elderly adults with certain chronic pulmonary, cardiovascular and metabolic conditions to receive influenza vaccine, vaccination rates remain low. Visits to subspecialists represent an important vaccination opportunity, but little is known regarding subspecialists' perceptions related to influenza vaccination. In February 2003, we conducted a cross-sectional mail survey of a random sample (N = 2,007) of board-certified cardiologists, endocrinologists and pulmonologists from the entire United States who provided outpatient care to adults aged 18-64 years, to assess their patterns of and attitudes toward administering influenza vaccine to high-risk, non-elderly patients. The overall response rate was 33%. Among 621 eligible respondents, 483 stocked influenza vaccine in their practice (Stockers) and 138 did not stock the vaccine (Non-Stockers). Pulmonologists were most likely to stock vaccine and strongly recommend vaccination; cardiologists were least likely. Among Stockers, barriers to vaccination varied by subspecialty. Among Non-Stockers, the most common factor in the decision to not stock vaccine was the perception that patients will receive the vaccine elsewhere. Most subspecialists who provide care to a large proportion of high-risk, non-elderly persons recommend influenza vaccination to some degree, particularly pulmonologists. To reduce missed opportunities overall, subspecialists should be encouraged to vaccinate patients who say that they plan to get the vaccine elsewhere. For cardiologists in particular, barriers to stocking influenza vaccine and recommending vaccination more strongly must be addressed. PMID- 18414062 TI - Dihydroartemisinin induces apoptosis in HL-60 leukemia cells dependent of iron and p38 mitogen-activated protein kinase activation but independent of reactive oxygen species. AB - Dihydroartemisinin (DHA), the main active metabolite of artemisinin derivatives, is one of the most effective anti-malarial analogs of artemisinin. In the current study, we found that DHA inhibited the proliferation of a panel of tumor cells originated from different tissue types. DHA effectively induced apoptosis in human promyelocytic leukemia HL-60 cells, which was accompanied with mitochondrial dysfunction and caspases activation. Further studies indicated that DHA-induced apoptosis was iron-dependent. Though DHA slightly elicited superoxide anion, these reactive oxygen species (ROS) contribute little to DHA-induced apoptosis in HL-60 cells. Moreover, DHA time-dependently activated mitogen activeted protein kinases (MAPKs) and specific inhibition of p38 MAPK, but not c Jun-NH2-terminal kinase (JNK) or extracellular signal-regulated kinase (ERK), abolished DHA-induced apoptosis, indicating that activation of p38 MAPK is required for DHA-induced apoptosis in HL-60 cells. Altogether, our data uncover that DHA induces apoptosis is dependent of iron and p38 MAPK activation but not ROS in HL-60 cells. PMID- 18414063 TI - Proteasome inhibitors increase tubulin polymerization and stabilization in tissue culture cells: a possible mechanism contributing to peripheral neuropathy and cellular toxicity following proteasome inhibition. AB - Bortezomib (Velcade((R))), a proteasome inhibitor, is approved by the FDA for the treatment of multiple myeloma (MM). While effective, its use has been hampered by peripheral neurotoxicity of unexplained etiology. Since proteasome inhibitors alter protein degradation, we speculated that proteins regulating microtubule (MT) stability may be affected after treatment and examined MT polymerization in cells by comparing the distribution of tubulin between polymerized (P) and soluble (S) fractions. We observed increased MT polymerization following treatment of SY5Y and KCNR [neuroblastoma], HCN2, and 8226 [MM] cells, using five proteasome inhibitors; the baseline proportion of total alpha-tubulin in 'P' fractions ranged from approximately 41-68%, and increased to approximately 55-99% after treatment. Increased acetylated alpha-tubulin, a post-translational marker of stabilized MTs, was observed in the neural cell lines HCN1A and HCN2 and this was sustained up to 144 hours after the proteasome inhibitor was removed. Cell cycle analysis of three cell lines after treatment, showed approximately 50-75% increases in the G(2)M phase. Immunofluorescent localization studies of proteasome inhibitor treated cells did not reveal microtubule bundles in contrast to paclitaxel treated, suggesting MT stabilization via a mechanism other than direct drug binding. We examined the levels of microtubule associated proteins and observed a 1.4-3.7 fold increase in the microtubule associated protein MAP2, in HCN2 cells following treatment with proteasome inhibitors. These data provide a plausible explanation for the neurotoxicity observed clinically and raise the possibility that microtubule stabilization contributes to cytotoxicity. PMID- 18414064 TI - Schizosaccharomyces pombe Orc5 plays multiple roles in the maintenance of genome stability throughout the cell cycle. AB - The six-subunit origin recognition complex (ORC) acts as a landing pad for factors that initiate DNA replication by binding to replication origins. In addition, ORC is involved in other processes such as transcriptional gene silencing and sister chromatid cohesion in Saccharomyces cerevisiae. However, whether these functions of ORC are specific to Saccharomyces cerevisiae or are shared by the ORC of other organisms is currently unclear. Analysis of two temperature-sensitive alleles of the fifth ORC subunit of Schizosaccharomyces pombe, orc5-H19 and orc5-H37, indicates that Orc5 of Schizosaccharomyces pombe has similar multiple functions to those of Orc5 of Saccharomyces cerevisiae. The orc5-H19 cells were defective in DNA replication initiation, and execution point analysis of this mutant revealed that ORC functions before metaphase to prepare for the initiation of replication in the next cell cycle. The orc5-H37 cells seemed to complete DNA synthesis but were arrested before entering M phase. In both mutants, the rads-chk1 checkpoint was activated to prevent mitosis, suggesting that this checkpoint pathway monitors the functional integrity of ORC. In addition, orc5-H37 cells showed premature separation of sister chromatids, which resulted in cell growth being dependent on the mad2-dependent spindle checkpoint. Consistently, this mutant showed a defect in the loading of Rad21, a cohesin component. Based on these observations, we propose that Orc5 has at least two distinct functions that can be separated genetically. Taken together, our results provide further support for the idea that ORC plays multiple functions during the cell cycle. PMID- 18414065 TI - Proteasome inhibitor MG-132 modifies coxsackie and adenovirus receptor expression in colon cancer cell line lovo. AB - The efficacy of adenovirus vector-based cancer gene therapy is controversial. Its uptake by cells in many cases requires the major receptor for adenoviruses, the coxsackievirus and adenovirus receptor (CAR). Low transduction is believed to be one of the main barriers as the expression of CAR on tumor cells is frequently reduced. Increasing CAR expression on tumor cells thus offers a promising opportunity for more effective adenovirus based treatment. Expression of CAR in 62 cases of colon tumor specimens were examined with immunohistochemistry. To modify the CAR expression, the effects of proteasome inhibitor MG132 on CAR expression of colon cancer cell lines were determined by flow cytometry, RT-PCR, and western blot. To evaluate adenovirus transfer, we further used rAd.EGFP, rAd.p53, and oncolytic adenovirus to infect target cells. The CAR expression was significantly decreased in colon carcinomas, both in primary tumors and lymphonode metastasis. Though the deregulation of CAR occurred in early disease and showed no relationship with TNM stage, when primary tumors are more than 5 cm in diameter, this deregulation becomes more frequent. More importantly, proteasome inhibitor MG-132 could enhance CAR expression in colon carcinoma cell line lovo, accompanied with enhanced adenovirus transfer, target gene expression, and oncolysis. These data provide a rational basis for evaluation of CAR expression in tumors and pretreatment with CAR conditioner prior to adenovirus vector-based gene therapy. PMID- 18414066 TI - Black and white female adolescents lose vitamin D metabolites into urine. AB - BACKGROUND: The black American population has a higher prevalence of salt sensitivity compared with the white American population. Dahl salt-sensitive rats, models of salt-induced hypertension, excrete protein-bound vitamin D metabolites into urine, a process that is accelerated during high salt intake. We tested the hypothesis that urinary vitamin D metabolite content and 25 hydroxyvitamin D (25-OHD) binding activity of black female adolescents would be greater than that of white female adolescents. METHODS: Female adolescents (11-15 years old, 11 black and 10 white) were fed low (1.3 g, 56 mmol/24 hours sodium) and high salt (3.86 g, 168 mmol/24 hours sodium) diets for 3 weeks in a randomized order cross-over study design. RESULTS: White and black adolescents had similar mean urinary vitamin D metabolite content (low salt, black versus white: 50 +/- 10 versus 58 +/- 17 pmol/24 hours; high salt, black versus white: 47 +/- 7 versus 79 +/- 16 pmol/24 hours). Mean urinary 25-OHD binding activities of the black and white adolescents did not significantly differ. Urinary 25-OHD binding activity of 10/11 black adolescents and 7/10 white adolescents was greater at week 3 of high salt intake than at week 3 of low salt intake (r = 0.50, P = 0.002, n = 17). Plasma 24,25-dihydroxyvitamin D concentrations of the white female adolescents were significantly higher than that of the black female adolescents (P < 0.001). CONCLUSION: Urinary loss of vitamin D metabolites may be one cause of low vitamin D status, in addition to low dietary intake and reduced skin synthesis. PMID- 18414067 TI - Continued smoking exacerbates but cessation ameliorates progression of early type 2 diabetic nephropathy. AB - PURPOSE: We tested the hypothesis that continued cigarette smoking exacerbates and its cessation ameliorates progression of the early nephropathy of type 2 diabetes mellitus (DM2) from microalbuminuria to macroalbuminuria. METHODS: We recruited 91 DM2 subjects with microalbuminuria, 39 nonsmokers and 52 smokers. Smokers underwent smoking cessation intervention with 11 of the 52 smokers quitting, yielding 3 groups: nonsmokers (NS, n = 39), continued smokers (S, n = 41), and quitting smokers (Quit, n = 11), all on angiotensin converting enzyme inhibition (ACEI), treated toward recommended BP and glycemic targets, and followed prospectively for 5 years. Subjects had yearly measurements of estimated glomerular filtration rate (eGFR) and albumin (mg)-to-creatinine (g) ratios (alb/cr) in spot morning urines. Comparison of changes in characteristics was done using analysis of variance, with all pair wise multiple comparison procedure at alpha = 0.05. RESULTS: Although average urine alb/cr was not different among groups at recruitment, 7 of the 41 S (17%) but none of the 50 NS or Quit progressed to macroalbuminuria (P < 0.003). eGFR decline rate was faster in S ( 1.79 +/- 0.35 mL/min/yr) than in NS or Quit (-1.30 +/- 0.43 and -1.54 +/- 0.37 mL/min/yr, respectively, P < 0.001). Multivariate analysis revealed smoking to be the only measured baseline factor that influenced eGFR decline rate (P < 0.041). CONCLUSION: Smoking exacerbates progression of early to advanced DM2 nephropathy and its cessation is an effective kidney-protective intervention in the early nephropathy of DM2. PMID- 18414068 TI - Hypovitaminosis D in African Americans residing in Memphis, Tennessee with and without heart failure. AB - BACKGROUND: Factors contributing to heart failure (HF) in African Americans (AA) are under investigation. Reduced 25(OH)D confers increased cardiovascular risk, including HF. METHODS: We monitored serum 25(OH)D, 1,25(OH)2D3, parathyroid hormone (PTH), and creatinine clearance in 102 AA residing in Memphis: 58 hospitalized with decompensated HF of >or=4 weeks in 34 (21 men; 53.3 +/- 1.8 years) or of 1 to 2 weeks in 24 (17 men; 49.6 +/- 2.4 years) and associated with a dilated cardiomyopathy and reduced ejection fraction (<35%); 19 outpatients with compensated HF (14 men; 52.6 +/- 2.7 years) with comparable ejection fraction; 16 outpatients (9 men; 55.4 +/- 2.9 years) with heart disease, but without HF; and 9 healthy volunteers (3 men; 35.8 +/- 3.5 years). RESULTS: Serum 25(OH)D 65 pg/mL was found in all AA with decompensated HF of >or=4 weeks (132.4 +/- 12.0 pg/mL) and 67% with 1 to 2 weeks duration (82.3 +/- 7.9 pg/mL), but only 11% with compensated HF (45.8 +/- 6.1 pg/mL), 12% without HF (29.6 +/- 5.4 pg/mL), and none of the volunteers (31.1 +/- 3.9 pg/mL). Creatinine clearance did not differ between patient groups. CONCLUSIONS: Hypovitaminosis D is prevalent amongst AA residing in Memphis, with or without HF. Elevations in serum PTH in keeping with secondary hyperparathyroidism are only found in AA with decompensated HF, where hypovitaminosis D and other factors are contributory. PMID- 18414069 TI - Catamenial diabetic ketoacidosis and catamenial hyperglycemia: case report and review of the literature. AB - BACKGROUND: Diabetic ketoacidosis (DKA), an acute and potentially life threatening complication of diabetes mellitus, is frequently the result of a precipitating illness or nonadherence to treatment; however, despite a thorough history, physical, radiologic, and laboratory examination, a specific cause or precipitating event cannot be identified in a significant number of cases. AIMS: To report 2 cases of recurrent DKA, and hyperglycemic crisis, associated to the menstrual cycle, and provide a review of the medical literature. METHODS: The clinic and hospital medical records of 2 subjects with type 1 diabetes mellitus (T1DM) and unexplained DKA are reviewed. An electronic MEDLINE search of relevant medical literature published from 1965 to 2007 was performed; additionally, the reference lists of the identified articles and other sources, such as textbook chapters and meeting abstracts, were reviewed for related publications. RESULTS: To date there have been 7 reported cases in the literature demonstrating an association between menstruation and DKA. The 2 new cases presented here highlight the potentially significant changes in glucose metabolism that may occasionally be observed during the late luteal and decidual phases of the menstrual cycle; moreover, these cases underscore the importance of a thorough medical and gynecologic history when evaluating a patient with DKA. CONCLUSION: Through unclear mechanisms, some women with diabetes mellitus demonstrate significant changes in glucose control around the time of their menses, including DKA. Accordingly, we propose that the terms catamenial DKA and catamenial hyperglycemia be used to refer to these disorders and that catamenial DKA be included in the differential diagnosis list of causes or precipitating events that can lead to DKA. PMID- 18414070 TI - Metastatic neuroendocrine tumor of unknown primary presenting as acute pancreatitis. AB - Neuroendocrine tumors of unknown primary site are rare. Among all the tumors of unknown primary, neuroendocrine tumors account for less than 5% of such cases. They are identified by immunohistochemical staining which is strongly positive for chromogranin, synaptophysin, or electron microscopy identification of neurosecretory granules. We present a case of metastatic poorly differentiated neuroendocrine tumor with no identifiable primary, presenting as acute pancreatitis, hypercalcemia, and disseminated bony metastasis. Such presentation has been rarely reported before. Although the nature of these tumors remains undefined, the diagnosis of poorly differentiated neuroendocrine carcinoma identifies a potentially treatable subgroup. PMID- 18414071 TI - 46, XX man with SRY gene translocation: cytogenetic characteristics, clinical features and management. AB - This report describes a well-masculinized 33-year-old man with infertility and primary hypogonadism in whom chromosomal evaluation revealed a 46, XX karyotype. This syndrome is a rare but important cause of hypergonadotropic hypogonadism in which the diagnosis can be delayed or missed. A review of the cytogenetic basis and clinical features is presented to raise awareness of this entity among clinicians and to emphasize the importance of appropriate laboratory testing when indicated. PMID- 18414072 TI - Positron emission tomography-positive squalene-induced lipoid pneumonia confirmed by gas chromatography-mass spectrometry of bronchoalveolar lavage fluid. AB - Squalene is a type of oil obtained from shark liver. We describe a 76-year-old man diagnosed with chronic exogenous lipoid pneumonia due to squalene. A chest CT scan revealed pulmonary consolidation with ground-glass opacities in the right upper lobe. Positron emission tomography (PET) revealed significant uptake of 2 deoxy-2-F-fluoro-d-glucose (FDG) and 3'-deoxy-3'-F-fluorothymidine (FLT). Bronchoalveolar lavage (BAL) fluid contained many lipid-laden macrophages, and a transbronchial lung biopsy specimen showed clusters of foamy macrophages in alveolar spaces and granulomatous lesions. In addition, the presence of squalene in the BAL fluid was confirmed by gas chromatography-mass spectrometry, leading to a diagnosis of squalene-induced lipoid pneumonia. To the best of our knowledge, this is the first report of squalene-induced lipoid pneumonia in which squalene itself was successfully detected. This case also suggests the possibility that lipoid pneumonia shows significant uptake in FDG-PET and FLT PET. PMID- 18414073 TI - Bronchobiliary fistula in a cirrhotic patient: a case report and review of the literature. AB - Bronchobiliary fistula is defined as the passage of bile in the bronchi. The presence of bronchobiliary fistula in patient with cirrhosis is extremely rare. Management of these fistulas is often very difficult and can be associated with high morbidity and mortality. We are presenting a patient with ethanol related cirrhosis and biliptysis in whom a diagnosis of bronchobiliary fistula was made. A review of the literature including diagnosis and management is performed. PMID- 18414074 TI - A patient with adult erythermalgia: evidence suggesting an autoimmune etiology. AB - We report the case of a woman with a combination of erythermalgia, idiopathic thrombocytopenic purpura, and vitamin B-12 deficiency with positive parietal cell antibodies. The patient was treated with intravenous administration of immunoglobulins together with small doses of prednisone, which resulted in an improvement in her platelet counts, rise in her vitamin B12 levels, and resolution of her painful discolored digits. These findings suggest an underlying autoimmune component to the development of erythermalgia. PMID- 18414075 TI - Isolated polyarticular septic arthritis: an atypical presentation of meningococcal infection. AB - We are presenting a case of a 19-year-old college student with sudden-onset, asymmetric polyarticular arthritis with Neisseria meningitidis 10 days after an acute upper respiratory infection consisting of fevers, chills, pharyngitis, and productive cough. Primary meningococcal septic arthritis is a rare entity. A majority of these cases present in a monoarticular fashion. The synovial fluid findings, although compatible with inflammatory arthritis, are not typical of septic arthritis. This entity, although rare, should be considered in the differential diagnosis of septic arthritis of large joints, especially since N. meningitiditis does not grow well on routine culture media. A literature review on the diagnosis, treatment, and prevention of primary meningococcal septic arthritis is presented. PMID- 18414076 TI - Community-acquired methicillin-resistant Staphylococcus aureus as a cause of Fournier's gangrene. AB - Community-acquired methicillin-resistant Staphylococcus aureus (CA-MRSA) has become an important pathogen in aggressive skin and soft-tissue infections in patients without risk factors for nosocomial infections. We describe a case of a previously healthy adult who developed fulminant sepsis from Fournier's gangrene caused by a strain of CA-MRSA containing the Panton-Valentine leukocidin genes. PMID- 18414077 TI - Sudden onset of diabetic ketoacidosis during consensus interferon therapy for chronic viral hepatitis C. AB - INTRODUCTION: Interferon (IFN)-alpha, used for the treatment of chronic hepatitis C, has rarely been associated with the development of diabetes. Consensus interferon (CIFN), which is used for nonresponders, has not been associated with this complication. We describe the first case of new onset diabetes in a patient on CIFN. CASE: A 50-year-old man with chronic hepatitis C and no prior diabetes was started on IFN-alpha. After 24 weeks with a detectable viral load, he was switched to CIFN. Four weeks later, he presented in diabetic ketoacidosis which resolved with appropriate therapy without any ascertainable etiology (glutamic acid decarboxylase negative). Within 9 months he was off insulin, and is off all diabetic medications with an HgA1c of 5.6% 2 years after the episode. PMID- 18414080 TI - Epidemiologists (of all people) should question journal impact factors. PMID- 18414081 TI - Impact factor: good reasons for concern. AB - This commentary emphasizes the importance of Hernan's contention that the impact factor's strong dependence on nonquality factors makes it utterly flawed as a way to evaluate quality of journals or papers. PMID- 18414082 TI - How come scientists uncritically adopt and embody Thomson's bibliographic impact factor? AB - The bibliographic impact factor (BIF) of Thomson Scientific is sometimes not a valid scientometric indicator for a number of reasons. One major reason is the strong influence of the number of "source items" or "articles" for each journal that the company chooses each year as BIF's denominator. The irresistible fascination with (and picturesque uses of) a construct as scientifically weak as BIF are simple reminders that scientists are embedded in and embody culture. PMID- 18414083 TI - The impact factor follies. PMID- 18414084 TI - Rise and fall of the Thomson impact factor. PMID- 18414086 TI - Measuring cognitive function with age: the influence of selection by health and survival. AB - BACKGROUND: Research into the pathophysiology of age-associated cognitive function and decline requires a valid estimate of cognitive function. However, this estimation can be grossly influenced by a selective loss to follow-up. METHODS: We investigated the influence of health selection on the estimated age associated cognitive function and decline by studying the effect on this estimation of study design and of the handling of multiple and missing data. We used linear regression analyses and linear mixed models to assess cognitive function from cross-sectional and longitudinal data. Repeated measures of cognitive function (assessed with dedicated neuropsychological tests) were carried out in 2 independent population-based cohort studies: the Rotterdam Study (3719 participants; mean age 71 years) and the Leiden 85-plus Study (369 participants; age 85 years). RESULTS: The effect of age on cognitive function was greater in cross-sectional analyses when all participants were included than when analyses were restricted to participants with repeated measurements. The decline in cognitive function over 4.6 years of follow-up was intermediate between the cross-sectional estimates from the total sample and from the restricted sample. Moreover, the estimated decline in cognitive function was larger when using a short follow-up than when using the complete follow-up over 5 years. The estimated decline using linear mixed models was similar to analyses including those with a complete follow-up over 5 years. CONCLUSION: Selection for health and survival results in better age-specific cognitive test scores and less cognitive decline. Statistical methods handling multiple and missing data do not fully correct for this bias. PMID- 18414087 TI - Cigarette smoking and dementia: potential selection bias in the elderly. AB - We conducted a systematic review of published prospective studies that estimated the association between smoking and the incidence of Alzheimer disease and dementia. The relative rate for smokers versus nonsmokers ranged from 0.27 to 2.72 for Alzheimer disease (12 studies) and from 0.38 to 1.42 for dementia (6 studies). The minimum age at entry (range: 55-75 years) explained much of the between-study heterogeneity in relative rates. We conjecture that selection bias due to censoring by death may be the main explanation for the reversal of the relative rate with increasing age. PMID- 18414085 TI - Air temperature and inflammatory responses in myocardial infarction survivors. AB - BACKGROUND: Temperature changes have been associated with increased cardiovascular risk, but the role of inflammatory markers in this relationship is not well understood. The objective of this study was to analyze the association between air temperature and C-reactive protein, interleukin-6 and fibrinogen in postmyocardial infarction patients. METHODS: In a multicenter panel study, the 3 inflammatory blood markers were measured repeatedly. In total, 5813 blood samples in 1003 subjects were collected in 6 European cities representing different climates. Data on patient characteristics and disease history were gathered at the baseline visit. Meteorologic data were obtained from the city-specific network stations. The association was analyzed using a semiparametric model with random patient effects. RESULTS: A 10 degrees C decrease in the 5-day-average of air temperature before the blood withdrawal was associated with a 4% increase in C-reactive protein (4.3% [95% confidence interval = 0.2% to 8.1%]). Correspondingly, an increase of interleukin-6 was observed for the same time window (3.3% [0.1% to 6.3%]) whereas fibrinogen showed an increase of 1.3% (0.2% to 2.4%) with a lag of 3 days. CONCLUSION: A decrease in air temperature, particularly the average temperature of the last 5 days, was associated with an increase in both C-reactive protein and interleukin-6, whereas fibrinogen seemed to react to temperature changes after 3 days. In susceptible patients this might lead to an additional risk for cardiovascular events and suggests a biologic mechanism for the observed seasonal variation in death from ischemic heart disease and stroke in the elderly. PMID- 18414088 TI - Socioeconomic disadvantage and acute coronary events: a spatiotemporal analysis. AB - BACKGROUND: The associations between socioeconomic disadvantage and ischemic heart disease are not well understood. We explore the relationship between socioeconomic factors and acute coronary events using spatiotemporal analysis. METHODS: We studied all deaths from acute myocardial infarction and hospital admissions for acute coronary syndrome and related revascularization procedures for the state of New South Wales, Australia, from 1996 through 2002. We used conditional autoregressive models to describe how characteristics of subjects' place of residence (socioeconomic disadvantage, proportion of the population of indigenous background, and metropolitan versus nonmetropolitan area) influenced admissions and mortality. RESULTS: There were 32,534 deaths due to acute myocardial infarction and 129,045 admissions for acute coronary syndrome. We found a relationship between increasing socioeconomic disadvantage and mortality (unadjusted relative risk for highest quartile of disadvantage relative to lowest = 1.40; 95% confidence interval = 1.27-1.54) as well as admissions (1.41; 1.28 1.55). After accounting for admission rates, socioeconomic disadvantage was associated with lower rates of angiography (0.75; 0.63-0.88) and interventional angiography (0.70; 0.56-0.85). After adjusting for socioeconomic disadvantage, areas with higher proportions of the population identified as indigenous had higher rates of admission and mortality, while residency in the state capital was associated with higher admission rates and more interventional angiography. After accounting for admission rates, the association of socioeconomic disadvantage with mortality was reduced. CONCLUSIONS: Socioeconomic disadvantage increases both the risk of acute coronary syndrome and related mortality. A contributing factor appears to be a reduced chance of receiving appropriate care. Regions with a higher proportion of indigenous residents show risk beyond the effects of general socioeconomic disadvantage, while residents of metropolitan communities had increased utilization of more recent interventions. PMID- 18414089 TI - Intelligent smoothing using hierarchical Bayesian models. AB - Hierarchical Bayesian modeling provides a flexible approach to modeling in multiparameter problems. Examples include disease mapping and spatiotemporal analysis, and multiple exposure modeling. A key feature of hierarchical Bayesian models is that prior expectations regarding model structure are embedded in a probability model that reflects uncertainty about the form of the structure that links analytical units (such as geographic areas). This results in posterior estimates that are compromises between raw data summaries and estimates that conform exactly to the prior model structure. The posterior estimates are more precise and generally have lower mean-squared error than traditional data summaries, and yet are not strictly constrained to follow a posited prior model form. PMID- 18414090 TI - Bone lead levels and blood pressure endpoints: a meta-analysis. AB - BACKGROUND: Previous reviews have shown increases in blood pressure and hypertension associated with increases in lead levels in blood. We performed a meta-analysis of the association of bone lead levels with systolic blood pressure, diastolic blood pressure, and hypertension using published data. METHODS: We searched Medline, Embase, and Toxline for epidemiologic studies on bone lead levels and blood pressure endpoints. We used inverse-variance weighted random-effects models to summarize the association of tibia or patella lead levels with blood pressure endpoints. RESULTS: We summarized data from 3 prospective studies and 5 cross-sectional studies. All studies measured lead levels in tibia bone and 3 studies measured lead levels in patella. For a 10 microg/g increase in tibia lead, the cross-sectional summary increases in blood pressure were 0.26 mm Hg for systolic (95% confidence interval = 0.02 to 0.50) and 0.02 mm Hg for diastolic (-0.15 to 0.19). The summary odds ratio for hypertension was 1.04 (1.01 to 1.07). For a 10 microg/g increase in patella lead, the summary odds ratio for hypertension was 1.04 (0.96 to 1.12). CONCLUSION: Systolic blood pressure and hypertension risk were associated with lead levels in tibia bone, but the magnitude of the summary estimates was small. These summary estimates, however, were based on published data and we could not evaluate nonlinear dose-response relationships, the relative contribution of bone and blood lead levels, or the influence of differences in study populations. A more detailed characterization of the association of bone lead levels and blood pressure endpoints would require a pooled analysis of individual participant data from existing studies. PMID- 18414091 TI - Prior assault and posttraumatic stress disorder after combat deployment. AB - BACKGROUND: Factors that make people vulnerable to or resilient against posttraumatic stress disorder (PTSD) following overwhelming stress are not well understood. The objective of this study was to prospectively examine the relation between prior assault and new-onset PTSD symptoms in a large US military cohort deployed in the wars in Iraq and Afghanistan. METHODS: Data on exposures and health outcomes were collected in the Millennium Cohort study at enrollment (July 2001 to June 2003) and follow-up (June 2004 to February 2006) from over 55,000 participants. Of these, 5324 were deployed in Iraq and Afghanistan, reported combat exposures, and were free of PTSD at baseline (881 women and 4443 men). We used multivariable logistic regression analysis to model the odds of new-onset PTSD in relation to prior assault. RESULTS: New-onset PTSD symptoms or diagnosis among deployers reporting combat exposures occurred in 22% of women who reported prior assault and 10% not reporting prior assault. Among men reporting prior assault, rates were 12% and 6%, respectively. Adjusting for baseline factors, the odds of new-onset PTSD symptoms was more than 2-fold higher in both women and men who reported assault prior to deployment. CONCLUSIONS: Prior assault appears to confer increased vulnerability for, rather than resilience against, PTSD symptoms among military professionals deployed to recent combat operations. PMID- 18414092 TI - A conversation with Henry Blackburn. Interviewed by Bernard L. Harlow. PMID- 18414093 TI - Edward S. Godfrey, Jr., founder of the first US Epidemiologic Society. PMID- 18414094 TI - Trends in autism spectrum disorder referrals. PMID- 18414096 TI - Measures of biological interaction and the STROBE statement. PMID- 18414097 TI - Suicide after breast augmentation. PMID- 18414099 TI - Screening for open angle glaucoma: systematic review of cost-effectiveness studies. AB - PURPOSE: To systematically review current evidence on the cost-effectiveness of screening strategies for open angle glaucoma (OAG). MATERIALS AND METHODS: Studies that reported both costs and outcomes of alternative screening strategies for OAG were identified by a highly sensitive search of electronic databases (eg, MEDLINE, EMBASE, NHS EED, HTA Database), last search December 2005. Data on costs regarding cases and years of visual impairment prevented, cases of blindness prevented, and cases of OAG detected were extracted. Incremental cost effectiveness ratios were calculated using data provided in the included studies. RESULTS: Four studies met the inclusion criteria. The latest of these was published in 1997. The screening tests and treatments reported in these studies are now not considered to be best practice. Furthermore, data were not reported in sufficient detail to reinterpret the results of the studies in terms of a common outcome measure. Finally, these studies suffered from methodologic weaknesses that further limit their usefulness for decision making. CONCLUSIONS: Currently, there is insufficient economic evidence on which to base recommendations regarding screening for OAG. New technologies, potentially suitable as screening devices, and new treatments are available. Further research, both in terms of economic models and conduct of clinical trials with concurrent economic evaluation, may help inform policy makers regarding cost effectiveness and acceptability of screening for OAG. PMID- 18414100 TI - Discrepancy of the intraocular pressure response between fellow eyes in one-eye trials versus bilateral treatment: verification with normal subjects. AB - PURPOSE: To compare the correlation of the fellow-eye's intraocular pressure (IOP) response in one-eye trials performed separately for each eye with that of bilateral treatment in normal subjects. METHODS: A one-eye trial with topical latanoprost applied once daily for 7 days was carried out in the right eye and then in the left eye of 41 normal subjects. Bilateral treatment was performed in a different set of 41 normal subjects. IOPs were measured at 3 time points on day 0 and on day 7. RESULTS: Latanoprost significantly reduced IOP of treated eyes in one-eye trials (2.8+/-1.6 and 2.7+/-1.6 mm Hg in the first and second trial, respectively) and in bilateral treatments (2.8+/-1.3 and 2.6+/-1.4 mm Hg in the right and left eye, respectively). Correlation of mean diurnal IOP reduction between 2 one-eye trials was poor (r2=0.102), even after subtracting the nontreated eye IOP fluctuations from the treated eye IOPs (r2=0.097), but that between fellow eyes in bilateral treatment was excellent (r=0.849). Correlation of baseline IOP at each time point between fellow eyes in one-eye trials and bilateral treatment (r2=0.729 to 0.949) was better than that in the same eye between 2 one-eye trials (r2=0.319 to 0.631). CONCLUSIONS: Fellow eyes in normal subjects showed a symmetrical IOP response to short-term bilateral treatment with latanoprost, although they did not respond symmetrically to one-eye trials performed separately for each eye. Poor correlation of IOP changes between 2 one eye trials may be caused by different IOP responsiveness to latanoprost at each trial, rather than asymmetrical IOP fluctuations. PMID- 18414101 TI - Effect of recording duration on the diagnostic performance of multifocal visual evoked potentials in high-risk ocular hypertension and early glaucoma. AB - PURPOSE: To evaluate the effect on diagnostic performance of reducing multifocal visual-evoked potential (mfVEP) recording duration from 16 to 8 minutes per eye. METHODS: Both eyes of 185 individuals with high-risk ocular hypertension or early glaucoma were studied. Two 8-minute mfVEP recordings were obtained for each eye in an ABBA order using VERIS. The first recording for each eye was compared against single run (1-Run) mfVEP normative data; the average of both recordings for each eye was compared against 2-Run normative data. Visual fields (VFs) were obtained by standard automated perimetry (SAP) within 22.3+/-27.0 days of the mfVEP. Stereo disc photographs and Heidelberg Retina Tomograph images were obtained together, within 24.8+/-50.4 days of the mfVEP and 33.1+/-62.9 days of SAP. Masked experts graded disc photographs as either glaucomatous optic neuropathy or normal. The overall Moorfields Regression Analysis result from the Heidelberg Retina Tomograph was used as a separate diagnostic classification. Thus, 4 diagnostic standards were applied in total, 2 based on optic disc structure alone and 2 others based on disc structure and SAP. RESULTS: Agreement between the 1-Run and 2-Run mfVEP was 90%. Diagnostic performance of the 1-Run mfVEP was similar to that of the 2-Run mfVEP for all 4 diagnostic standards. Sensitivity was slightly higher for the 2-Run mfVEP, whereas specificity was slightly higher for the 1-Run mfVEP. CONCLUSIONS: If higher sensitivity is sought, the 2-Run mfVEP will provide better discrimination between groups of eyes with relatively high signal-to-noise ratio (eg, early glaucoma or high-risk suspects). But if higher specificity is a more important goal, the 1-Run mfVEP provides adequate sensitivity and requires only half the test time. Considered alongside prior studies, the present results suggest that the 1-Run mfVEP is an efficient way to confirm (or refute) the extent of VF loss in patients with moderate or advanced glaucoma, particularly in those with unreliable VFs, including malingering or other "functional" forms of VF loss. PMID- 18414102 TI - Detection of early glaucoma with optical coherence tomography (StratusOCT). AB - PURPOSE: To evaluate the performance of optical coherence tomography (StratusOCT) for discriminating eyes with early glaucoma from normal eyes. METHODS: Thirty eyes with established early glaucomatous visual field defects (EGVF group), 30 eyes with evidence of early glaucomatous optic neuropathy with normal standard achromatic perimetry [early glaucoma by disc (EGD)], and 33 age-matched normal eyes with good quality StratusOCT nerve fiber layer (NFL) images were enrolled. Average NFL thickness and NFL thickness at quadrants and sectors, areas under receiver operator characteristic curves, and sensitivities at 80% and 90% specificity were evaluated. RESULTS: The average (+/-SD) mean deviation in the EGVF group was -3.4 (+/-1.7) dB. Receiver operator characteristic curves showed areas under the curve (AUC) for NFL thickness in the superior quadrant (AUC=0.75+/-0.07) and in the inferior quadrant (AUC=0.94+/-0.03) to be the best StratusOCT parameters for discrimination of normal controls from EGD and EGVF eyes, respectively. The best parameter for detection of EGD eyes at 80% and 90% specificities was NFL thickness at superior quadrant (51% and 36% sensitivities, respectively). The best parameter for detection of EGVF eyes at 80% and 90% specificities was NFL thickness in the inferior quadrant (90% and 87% sensitivities, respectively). CONCLUSIONS: Optical coherence tomography (StratusOCT) showed good sensitivity and specificity in a group of glaucoma patients with early visual field loss. In patients with normal visual fields in whom the optic disc appeared glaucomatous to glaucoma specialists, half were confirmed to have StratusOCT findings consistent with damage from glaucoma. PMID- 18414103 TI - Contribution of CYP1B1 mutations and founder effect to primary congenital glaucoma in Mexico. AB - PURPOSE: The frequency of primary congenital glaucoma (PCG)-causing CYP1B1 mutations varies importantly among distinct populations, ranging from 20% in Indonesians and Japanese to about 100% among the Saudi Arabians and Slovakian Gypsies. Thus, the molecular characterization of large groups of PCG from different ethnic backgrounds is important to establish the actual CYP1B1 contribution in specific populations. In this work, the molecular analysis of the CYP1B1 gene in a group of Mexican PCG patients is reported. MATERIAL AND METHODS: Thirty unrelated Mexican patients fulfilling the clinical criteria for PCG were included. Two cases were familial and with proven consanguinity, originating from distinct regions of the country. Polymerase chain reaction amplification and direct automated sequencing of the CYP1B1 coding region was performed in each participating subject. RESULTS: An identical pathogenic CYP1B1 mutation was demonstrated in 2 unrelated PCG subjects. The mutation consisted of a homozygous G to A transition at nucleotide position 1505 in exon 3, which predicted a substitution of glutamic acid for lysine at residue 387 of the protein (E387K). In the remaining 28 PCG subjects, no deleterious mutations were identified. Both subjects with the E387K mutation shared a same haplotype for 5 CYP1B1 intragenic single nucleotide polymorphisms, indicating a common origin of the allele. CONCLUSIONS: Mexican patients with PCG are rarely (less than 10%) due to CYP1B1 mutations. Available data indicate that most of the non-Brazilian Latin American PCG patients investigated to date are not due to CYP1B1 defects. Populations with low incidence of CYP1B1 mutations are appropriate candidates for the identification of novel PCG-causing genes. PMID- 18414104 TI - Three-dimensional anterior segment optical coherence tomography of filtering blebs after trabeculectomy. AB - PURPOSE: To evaluate trabeculectomy blebs by using 3-dimensional anterior segment optical coherence tomography (OCT). METHODS: We prospectively examined 4 eyes of 4 patients who developed filtering blebs after trabeculectomy. A 1310-nm high speed OCT prototype was used to image the 3-dimensional structure of the filtering blebs. RESULTS: The 3-dimensional structure of the filtering blebs was clearly observed in the OCT images. Three types of filtering blebs were observed: diffuse blebs in 2 eyes, an encapsulated bleb in 1 eye, and a nonfunctioning cystic bleb in 1 eye. The volume of each bleb was 9.97, 1.10, 0.76, and 0.88 mm3, respectively. En-face OCT images clearly showed the aqueous outflow channels at the margins of the scleral flaps. CONCLUSION: Three-dimensional OCT allows objective and noninvasive assessment of filtering blebs after trabeculectomy. PMID- 18414105 TI - Evaluation of a modified protocol for selective laser trabeculoplasty. AB - PURPOSE: To compare the intraocular pressure (IOP) response to a modified protocol for selective laser trabeculoplasty (SLT) to standard protocols for SLT and argon laser trabeculoplasty (ALT). MATERIALS AND METHODS: A retrospective study of 318 eyes of 284 patients diagnosed with either primary open angle, pigmentary or pseudoexfoliation glaucoma who underwent laser trabeculoplasty from September 1997 to September 2005. One hundred and two patients, who underwent a modified SLT protocol with 100 overlapping laser spots over 180 degrees of trabecular meshwork were compared with 89 patients who received SLT with 100 nonoverlapping spots over 360 degrees and another 127 patients who received ALT with 50 spots over 180 degrees. IOPs were measured at baseline and postoperatively at 1 hour, 6 weeks, 4 months, and 14 months. Regression models, based on the observed data, were used to predict the fall in IOP in the 3 groups, controlling for differences in baseline pressure. RESULTS: The IOP response to overlapping SLT was significantly worse than to nonoverlapping SLT or ALT, both of which had similar responses. Baseline IOP was the only preoperative factor that predicted response to ALT (P<0.0001) and nonoverlapping SLT (P=0.0019) at all follow-up times. There were no statistically significant predictive factors for IOP reduction in the overlapping SLT group. CONCLUSIONS: Overlapping application of SLT results in a poorer IOP response compared with ALT and nonoverlapping SLT. PMID- 18414106 TI - Relationship between Humphrey 30-2 SITA Standard Test, Matrix 30-2 threshold test, and Heidelberg retina tomograph in ocular hypertensive and glaucoma patients. AB - PURPOSE: To evaluate the relationship between global indices of Humphrey standard automated perimetry (SAP, 30-2 SITA standard test), Humphrey Matrix frequency doubling technology (FDT, 30-2 threshold test), and Heidelberg Retina Tomograph (HRT II) parameters and measure the level of agreement among these 3 tests in classifying eyes as normal or abnormal. METHODS: The study included 1 eye of 29 ocular hypertensive and 56 glaucoma patients with a mean age of 60.9+/-10.5 years. All subjects had reliable visual fields and HRT measurements performed within a 2-week period. The eyes were classified as normal/abnormal according to visual field criteria and Moorfields regression analysis (MRA). Correlations between visual field indices (mean deviation and pattern standard deviation) and HRT parameters were analyzed using Spearman correlation coefficient (r) and the agreement between the tests in classifying eyes was defined with kappa value. RESULTS: FDT Matrix mean deviation and pattern standard deviation parameters were found to be highly correlated with those of SAP (r=0.66 and 0.69, respectively). Visual field indices showed statistically significant correlations with cup area, rim area, cup/disc (C/D) area, linear C/D, cup shape, mean retinal nerve fiber layer thickness and retinal nerve fiber layer area parameters (P<0.05). Fifty eight patients (68.2%) had abnormal results at least with 1 of the tests and 21 subjects (24.7%) had abnormal results with all 3 tests. The kappa values were 0.6 for SAP and Matrix (P<0.001), 0.33 for SAP and MRA (P=0.002), and 0.31 for Matrix and MRA (P=0.004). CONCLUSIONS: FDT Matrix results are highly comparable with SAP in the assessment of glaucoma. Visual field global indices show statistically significant, but low-moderate correlations with most of the HRT parameters. The agreement among MRA and visual fields for abnormality is fair. Either HRT or visual fields may show the first evidence of glaucomatous damage; therefore, the combination of optic nerve head parameters and visual field results could improve glaucoma diagnosis and follow-up. PMID- 18414108 TI - Duration of IOP reduction with travoprost BAK-free solution. AB - PURPOSE: To compare the duration of action of travoprost ophthalmic solution 0.004% (Travatan Z) formulated without benzalkonium chloride (BAK) to travoprost ophthalmic solution 0.004% formulated with BAK (Travatan). METHODS: This was a prospective, randomized, double-masked study. Patients with open-angle glaucoma or ocular hypertension were randomized to receive 2 weeks of once-daily therapy with travoprost BAK-free or travoprost with BAK. Patients received the last dose of medication on day 13 and then intraocular pressure (IOP) was assessed every 12 hours for 60 hours. Statistical analysis included change in IOP from baseline for each group and comparison of mean IOP between groups. RESULTS: Of the 109 patients enrolled, 106 patients completed the study. Untreated mean baseline IOP at 8 AM was 26.9 mm Hg in the travoprost BAK-free group and 27.1 mm Hg in the travoprost with BAK group. At 12, 24, 36, 48, and 60 hours after the last dose, mean IOP in the travoprost BAK-free group was 18.7, 17.2, 19.5, 18.7, and 20.8 mm Hg, respectively; whereas mean IOP in the travoprost with BAK group was 18.5, 16.8, 19.7, 18.0, and 20.8 mm Hg, respectively. Mean IOP at all time points after the last dose of medication was >6 mm Hg lower than the 8 AM baseline in both groups. Between-group differences were within +/-0.6 mm Hg at all postdose time points. There were no statistically significant differences between the 2 treatment groups at baseline or at any postdose time point. Drug-related side effects were uncommon, mild in intensity, and comparable between groups. CONCLUSIONS: Travoprost without BAK has similar IOP-lowering efficacy and safety compared with travoprost preserved with BAK. Both formulations of travoprost have a prolonged duration of action, with statistically and clinically significant reductions from baseline persisting up to 60 hours after the last dose. PMID- 18414107 TI - Bimatoprost/timolol fixed combination: a 3-month double-masked, randomized parallel comparison to its individual components in patients with glaucoma or ocular hypertension. AB - PURPOSE: To evaluate the safety and efficacy of a fixed combination (FC) of bimatoprost (BIM) and timolol (TIM) compared with each of the active components for 3 months. PATIENTS AND METHODS: Two double-masked, randomized, multicenter parallel studies of FC (once-daily, mornings), BIM (once-daily, evenings), or TIM (twice-daily) were conducted in 1061 patients with glaucoma or ocular hypertension. RESULTS: Mean diurnal decreases from baseline intraocular pressure (IOP) at month 3 were 8.1, 7.9, and 6.4 mm Hg for the FC, BIM, and TIM groups, respectively. The proportion of patients with a mean diurnal percent reduction from baseline in IOP of more than 20% across all visits was 81.8% (436/533), 72.1% (191/265), and 49.8% (131/263) for the FC, BIM, and TIM groups, respectively (P<0.001 for FC vs. BIM and FC vs. TIM). The proportion of patients achieving an IOP of less than 18 mm Hg at all time points was 39.2% (209/533), 28.7% (76/265), and 12.2% (32/263) for the FC, BIM, and TIM groups, respectively (P=0.003 for FC vs. BIM, and P<0.001 for FC vs. TIM). The most commonly reported treatment-related adverse event was conjunctival hyperemia, with the greatest incidence in BIM (38.5%, 102/265), followed by FC (22.7%, 121/533, P<0.0001 vs. BIM) and TIM (6.8%, 18/263; P<0.001 vs. FC). CONCLUSIONS: FC was statistically significantly more effective than BIM or TIM for most comparisons, and safer than BIM with respect to common ocular adverse events. FC represents a convenient, therapeutic advantage over separate bottles. PMID- 18414109 TI - No association between Helicobacter pylori infection or CagA-bearing strains and glaucoma. AB - BACKGROUND AND PURPOSE: Accumulating evidence indicates that a variety of infections contribute to the pathogenesis of glaucoma. The role of Helicobacter pylori infection in glaucoma is controversial. DESIGN: Prospective, population based study. PARTICIPANTS: Patients with various types of glaucoma and a control group of patients with cataract. METHODS: We evaluated seropositivity to H. pylori and to its cytotoxin-associated gene A (CagA) product in patients with various types of glaucoma and compared the findings to those of a control group of patients with cataract. RESULTS: H. pylori infection and CagA seropositivity were detected in 31/51 (60.8%) and 26/51 (51%) glaucoma patients compared with 22/36 (61.1%) and 19/36 (52%) control patients, respectively (P=0.88, 0.67, not significant). Similar rates of H. pylori infection and CagA-positive strain were found in all glaucoma subgroups, and none of them was statistically different from those of controls. CONCLUSIONS: Neither H. pylori infection nor seropositivity for virulent CagA-bearing H. pylori strains have significant association with the occurrence of glaucoma of any type. PMID- 18414110 TI - Characteristics of patients who dropout from a glaucoma clinic. AB - PURPOSE: To find the dropout rate and identify the clinical characteristics of patients who drop out in the first year of follow-up from a glaucoma clinic. DESIGN: Descriptive hospital-based study at a tertiary hospital eye department. METHODS: Clinical characteristics of consecutive patients newly diagnosed with glaucoma who dropped out (n=452) were compared with patients who did not drop out (n=295) within 12 months. RESULTS: The rate of dropout from follow-up was 60.5% within 1 year; 43.1% of the study group dropped out after their first follow-up visit. The dropout rate was high in all age groups, but higher in the age groups 21 to 30 years, 41 to 50 years, and over 70 years. Males had a higher dropout rate than females (78.6% vs. 34.5%). Dropout rate was higher among those with mild/moderate glaucoma than those with severe disease (88.2% vs. 37.2%); those who lived further away from the hospital than those who lived nearer to the hospital (72.5% vs. 40.8%), those who were referred from screening clinics for nonblinding eye disease compared with those referred because of a blinding eye disease (72.2% vs. 58.9%). More patients (63.8%) unsure of their family eye disease history dropped out, compared with 34.3% of those with positive family history of glaucoma and other potentially blinding diseases. More patients who had no systemic disease dropped out, than those with systemic disease (54.6% vs. 39.6%); whereas patients on 2 medications or more had a higher dropout rate than those on less than 2 medications (68.1% vs. 52.1%). Of the study factors, those that were statistically significantly associated with dropping out of follow-up from the glaucoma clinic were age, sex, place of domicile, diagnosis at referral, severity of disease, family history, and polydrug use. CONCLUSIONS: The dropout rate from this glaucoma clinic in the first year was high (60.5%). Patients who were more likely to dropout were younger patients, male, those who travelled far distances to the clinic, those with mild to moderate glaucoma, those with no family history of blinding eye diseases, and patients taking 2 or more eyedrops. Patients who seem to perceive their problems as not serious dropped out of follow up. These findings have great implications in planning future studies and intervention to improve the follow-up of glaucoma patients in the study area. PMID- 18414111 TI - Comparison of ocular hypotensive effect and safety of brinzolamide and timolol added to latanoprost. AB - PURPOSE: To compare the ocular hypotensive effect and safety of brinzolamide and timolol added to latanoprost monotherapy. METHODS: In prospective randomized fashion, we evaluated the ocular hypotensive effect and safety of brinzolamide or timolol in 1 eye of 32 patients with primary open-angle glaucoma, normal-tension glaucoma, or ocular hypertension who had been treated with latanoprost for more than 1 month. Intraocular pressure (IOP), blood pressure, and pulse were measured before and at 4, 8, and 12 weeks. Corneal endothelial cell density was measured at baseline and at 12 weeks. RESULTS: The IOP was 17.8+/-1.7 mm Hg (mean+/-SD) before the addition of brinzolamide (n=15) and 15.7+/-2.1 mm Hg at 12 weeks (P<0.01). In comparison, the IOP was 18.5+/-3.7 mm Hg before the addition of timolol (n=15) and 15.8+/-3.2 mm Hg at 12 weeks (P<0.01). Both brinzolamide and timolol significantly decreased IOP at 12 weeks, by a mean of 2.0 mm Hg and mean 2.7 mm Hg, respectively, and were more effective than latanoprost alone (P<0.01), but there were no significant differences between the drugs and no significant differences in corneal endothelial cell density and blood pressure before and after addition of either drug. At 12 weeks, pulse was decreased in patients receiving timolol (P<0.01). As systemic adverse events, there was one instance of malar flushing after brinzolamide addition and episodes of chest discomfort after timolol addition in 1 patient. Ocular adverse events were slight. CONCLUSIONS: Brinzolamide and timolol added to latanoprost have similar ocular hypotensive effects and safety in primary open-angle glaucoma, normal-tension glaucoma, or ocular hypertension. PMID- 18414112 TI - Endoscopic and transscleral cyclophotocoagulation for the treatment of refractory glaucoma. AB - PURPOSE: Cyclophotocoagulation (CPC) is traditionally used in cases of glaucoma that are refractory to medical and surgical therapy. The goal of this review is to provide an update on the methods, efficacy, complications, indications, and histopathology of transscleral and endoscopic CPC. METHODS: A literature review was conducted for transscleral and endoscopic CPC. Relevant studies were included for evaluation of the procedures. RESULTS: For transscleral CPC (TCP), there is a wide range of success rates reported in the literature, depending on energy settings, follow-up period, and definitions of success. Repeat TCP is often required. Serious complications have included significant vision loss, hypotony, and phthisis. TCP has mostly been performed in very severe forms of glaucoma, in eyes with limited visual potential, or after filtration surgery has failed, although more recently TCP has been used as a primary surgery in eyes with good visual potential. There are more limited published results on endoscopic CPC (ECP), which have demonstrated overall good success. Complication rates are relatively low with ECP, however, large studies with long-term follow-up are lacking. ECP has also been used in difficult, refractory cases, but often used earlier when combined with cataract surgery. CONCLUSIONS: Both TCP and ECP are effective surgeries with potential for serious complications. Recent studies suggest they may be used increasingly as the primary surgery for various stages of glaucoma. PMID- 18414114 TI - Atypical retinitis pigmentosa masquerading as primary open angle glaucoma. PMID- 18414116 TI - Prevalence of open-angle glaucoma, glaucoma suspect, and ocular hypertension in thyroid-related immune orbitopathy. PMID- 18414117 TI - Uncontrolled intraocular pressure after endoscopic cyclophotocoagulation. PMID- 18414119 TI - Big pharma and American psychiatry. PMID- 18414120 TI - The accuracy of psychiatric medication advertisements in medical journals. AB - Psychiatric medications are frequently advertised in medical journals, yet no study has addressed the veracity of claims made in these advertisements. The present study examined the accuracy of 69 medical journal advertisements for psychiatric medications and the availability of sources cited in these advertisements. Just over half of claims made in advertisements (50.2%) provided no attainable source that could be used to check the veracity of the claim. When sources were attained, they supported the cited claims 65% of the time (95% CI: 61.0-69.1). Claims regarding the efficacy of medications were only supported by obtained cited sources on 53.2% of occasions (95% CI: 46.2-60.2). Attempts to obtain cited data on file from sponsoring drug companies were rarely successful. Given the relatively poor empirical substantiation of claims made in medical journal psychiatric drug advertisements and that most claims provided no attainable sources, increased regulation of such advertising is warranted. PMID- 18414121 TI - Assessment of medication adherence in patients with schizophrenia: the Achilles heel of adherence research. AB - Measuring medication adherence in patients with schizophrenia is difficult and lacks a gold standard. Consequently, a great number of different methods and instruments have been proposed. Although it has been assumed that they all measure medication adherence, this study demonstrates that instruments differ significantly. Using data from an international multisite study (N = 329), we found that 3 questionnaires, designed to measure medication adherence and typical for instruments used in studies in patients with schizophrenia, do not agree in labeling patients as nonadherent. Further, they seem not to measure the same trait, are related to different established risk factors of nonadherence, and are only weakly related to these established risk factors of nonadherence. If these results are representative of the validity of other measures used in adherence research, this may have serious consequences for the interpretation of, and explanations for discrepancies found in the literature. Researchers should be aware of this problem and continue to combine objective and subjective methods in the hope of increasing the reliability and validity of measures of adherence. PMID- 18414122 TI - Nonverbal behavior during standardized interviews in patients with schizophrenia spectrum disorders. AB - Several studies have consistently shown that patients with schizophrenia or schizophrenia spectrum disorders (SSD) can be distinguished from normal controls on the basis of their nonverbal behavior during standardized interviews, with considerable interactions between negative symptoms and poor facial expressivity. However, most studies have examined unmedicated patients, and gender of both interviewer and interviewee has not been taken into account. In this study we assessed the nonverbal behavior of male and female patients with SSD who were receiving second-generation antipsychotic medication (SGA) using the Ethological Coding System for Interviews (Troisi, 1998). In addition, we used a novel 5 factor model of the Positive and Negative Symptom Scale (PANSS, van der Gaag et al., 2006) to correlate nonverbal behavior with standard psychopathology ratings. Our findings strongly resembled results of previous studies into nonverbal behavior of patients with SSD, despite differences in cultural backgrounds and gender of the interviewer. Negative symptoms were inversely correlated with several of the nonverbal behavioral dimensions. Medication dose did not correlate with any one of the behavioral or psychopathological measures. Patients with SSD make less use of their nonverbal behavioral repertoire compared with controls, independent of antipsychotic treatment. Culture-specific nonverbal expressivity seems to play an additional (minor) role in distinguishing patients from healthy controls. PMID- 18414123 TI - The development of valid subtypes for depression in primary care settings: a preliminary study using an explanatory model approach. AB - A persistent theme in the debate on the classification of depressive disorders is the distinction between biological and environmental depressions. Despite decades of research, there remains little consensus on how to distinguish between depressive subtypes. This preliminary study describes a method that could be useful, if implemented on a larger scale, in the development of valid subtypes of depression in primary care settings, using explanatory models of depressive illness. Seventeen depressed Hispanic patients at an inner city general practice participated in explanatory model interviews. Participants generated illness narratives, which included details about symptoms, cause, course, impact, health seeking, and anticipated outcome. Two distinct subtypes emerged from the analysis. The internal model subtype was characterized by internal attributions, specifically the notion of an "injured self." The external model subtype conceptualized depression as a reaction to life situations. Each subtype was associated with a distinct constellation of clinical features and health seeking experiences. Future directions for research using explanatory models to establish depressive subtypes are explored. PMID- 18414124 TI - Differential item functioning between ethnic groups in the epidemiological assessment of depression. AB - A potential explanation for the finding that disadvantaged minority status is associated with a lower lifetime risk for depression is that individuals from minority ethnic groups may be less likely to endorse survey questions about depression even when they have the same level of depression. We examine this possibility using a nonparametric item response theory approach to assess differential item functioning (DIF) in a national survey of psychiatric disorders, the National Comorbidity Survey. Of 20 questions used to assess depression symptoms, we found evidence of DIF in 3 questions when comparing non Hispanic blacks with non-Hispanic whites and in 3 questions when comparing Hispanics with non-Hispanic whites. However, removal of the questions with DIF did not alter the relative prevalence of depression between ethnic groups. Ethnic differences do exist in response to questions concerning depression, but these differences do not account for the finding of relatively low prevalence of depression among minority groups. PMID- 18414126 TI - Distress and affective dysregulation in patients with borderline personality disorder: a psychophysiological ambulatory monitoring study. AB - Borderline personality disorder (BPD) is characterized by enduring psychological distress and affective dysregulation. Several models have linked both phenomena, but are lacking empirical support. To investigate the relation between psychological distress and components of affective dysregulation (especially inability to label emotions, conflictive emotions, and physiological hyperarousal), we repeatedly assessed these components using a 24-hour ambulatory monitoring approach in a group of 50 BPD patients and 50 healthy controls. Hierarchical linear model analyses identified a clear relation between inability to label emotions and distress in the BPD group (p = 0.0009) but not across all subjects (p = 0.6492). Conflictive emotions were related to psychological distress in both groups (p < 0.0001). This relation is, however, most pertinent to the BPD group who experienced conflicting emotions more frequently. Physiological arousal (heart rate) was related to distress in both groups. Our empirical findings emphasize training in labeling emotions and distress tolerance interventions in treatment for BPD. PMID- 18414125 TI - Cross-cultural study of conviction subtype Taijin Kyofu: proposal and reliability of Nagoya-Osaka diagnostic criteria for social anxiety disorder. AB - Conviction subtype Taijin-Kyofu (c-TK) is a subgroup of mental disorder characterized by conviction and strong fear of offending others in social situations. Although the concept of c-TK overlaps with that of social anxiety disorder (SAD), patients with c-TK often may not be diagnosed as such within the current Diagnostic and Statistical Manual of Mental Disorders (DSM)-IV criteria. We propose the Nagoya-Osaka criteria to amend this situation. This study examined the cross-cultural interrater reliability of the proposed criteria. Eighteen case vignettes of patients with a variety of complaints focused around social anxieties were collected from 6 different countries, and diagnosed by 13 independent raters from various nationalities according to the original DSM-IV and the expanded criteria. The average agreement ratio for the most frequent diagnostic category in each case was 61.5% with DSM-IV and 87.6% with the modified DSM-IV with Nagoya-Osaka criteria (p < 0.001). These findings indicate that the Nagoya-Osaka criteria for SAD can improve interrater reliability of SAD. PMID- 18414127 TI - Psychometric properties of the inventory of gambling situations with a focus on gender and age differences. AB - Environmental, interpersonal, and psychological factors contribute to gambling, but no standardized instrument for assessing high-risk gambling situations exists. This study evaluates the factor structure of the Inventory of Gambling Situations (IGS), a newly developed measure to examine situations that precipitate gambling. Pathological gamblers (n = 233) beginning a treatment study completed the IGS. Principal component analysis revealed that a 5-factor solution best fit the data, representing items related to Negative Affect, Luck and Control, Positive Affect, Social Situations, and Gambling Cues. Overall, Cronbach alpha was 0.960, ranging from 0.820 to 0.935 for the 5 factors. IGS scores correlated with other indices of gambling problems. Women scored higher on the Gambling Cues factor than men, and younger respondents, regardless of gender, scored higher on the Luck and Control factor than older respondents. The IGS may assist in better understanding situations in which pathological gamblers wager, and individual differences with respect to these situations. PMID- 18414128 TI - Patients' and psychiatrists' perceptions of clinical decisions during schizophrenia treatment. AB - Shared decision making is advocated for patients with schizophrenia. However, there is limited knowledge as to which events are actually considered to be decisions by psychiatrists and patients. Semistructured interviews with regard to clinical decisions of the preceding week were performed with psychiatrists and inpatients with schizophrenia. There was good correspondence between patients and psychiatrists regarding decisional topics but poor correspondence regarding individual decisions. Medication issues were the most prominent, but other topics were also frequently cited. Not being included in decisions was associated with patients' desire to make the decisions differently. Patients treated involuntarily felt more often that they were not included in decisions and wanted to make different decisions. Thus, many patients do not feel involved in treatment decisions and are at the risk of noncompliance since they state that they would have made decisions differently from their psychiatrists. This is especially true of those being treated involuntarily. PMID- 18414129 TI - Measurement of insight in patients with bipolar disorder: are self-rated scales developed for patients with schizophrenia applicable? AB - Our aim was to study if the Birchwood Insight Scale has acceptable psychometric properties when used for patients with bipolar disorders. Patients with schizophrenia (n = 101), bipolar I (n = 57), and bipolar II disorder (n = 37) completed the self-report scale. The items form 3 subscales, awareness of illness, relabeling of symptoms, and need for treatment. The total scale had good internal consistency for patients with schizophrenia, fairly good for bipolar I, but poor for bipolar II disorder. On subscale level the internal consistency was mostly marginal to poor for all patient groups. The level of insight was similar in schizophrenia and bipolar I disorder. The psychometric properties for the insight scale were poorer in bipolar disorders than in schizophrenia, and the scale did not work for patients with bipolar II disorder. This suggests a cautious use of the scale across different diagnostic groups. PMID- 18414130 TI - Ranking of outcome domains for use in real-time outcomes feedback laboratory by patients with schizophrenia. AB - The purpose of this study was to prioritize real-time outcomes from the perspective of patients with schizophrenia. Thirty-eight patients were interviewed and ranked 15 outcome domains according to their preference for communicating these outcomes with their mental health provider in the context of a real-time outcomes feedback laboratory. Subjects with schizophrenia ranked overall physical health as the most important outcome domain that they wanted to communicate to their mental health provider in the context of a real-time outcomes feedback laboratory. The ranking of physical health as the most important outcome domain was true for subjects regardless of total psychosis symptom severity, positive symptom severity, age, or level of insight. Outcomes feedback interventions for patients with schizophrenia should include physical health problems in addition to the usual mental health parameters. PMID- 18414131 TI - The psychological impact of a catastrophic earthquake: a retrospective study 50 years after the event. AB - Aim of the present study was to retrospectively assess the impact of a catastrophic earthquake in a sample of 121 survivors, 50 years after the event. Mean age +/- SD of the responders was 72.2 +/- 6.1 years. The majority of the victims (78%) acknowledged a strong overall impact of the earthquake on their lives, and almost all of them had intense recollection of the event at its anniversary. The most frequent symptom during the 6 months after the earthquake was persistent remembering or "reliving" of the event; women had considerably more often recurrent dreams of the earthquake and distress than did men. Women and young adults at the time of the earthquake appear to be the most vulnerable groups regarding the psychological effects of the event. PMID- 18414135 TI - Brain hyperthermia after traumatic brain injury does not reduce brain oxygen. AB - OBJECTIVE: Hyperthermia can exacerbate outcome after traumatic brain injury (TBI). In this study, we examined the relationship between brain temperature (BT) and core body temperature and the relationship between BT and brain tissue oxygen (BtO2) to determine whether hyperthermia adversely affects BtO2. METHODS: Seventy two patients (mean age, 41 +/- 19 years) admitted to a Level I trauma center after TBI were retrospectively identified from a prospective observational database. Intracranial pressure (ICP), BT, and BtO2 were recorded continuously. Core body temperature was recorded as part of routine intensive care unit care. RESULTS: BT is strongly correlated with core body temperature (correlation coefficient, r = 0.92) over a wide range. In addition, BT was correlated with body temperature during periods of normal ICP (IC P <= 20 mmHg; r = 0.87) and transiently elevated ICP (ICP range 21-63 mmHg; r = 0.94). During periods of brain normothermia (BT < 38.1 degrees C), the average BtO2 was 36.3 +/- 22.9 mmHg. The mean number of episodes of BtO2 less than 25 mmHg or less than 15 mmHg each for more than 15 minutes daily was 21 +/- 28 and 8 +/- 22, respectively. The mean BtO2 (37.2 +/- 16.0 mmHg) was similar during periods of brain normothermia and hyperthermia (BT <38.1 degrees C). When the periods of brain tissue hyperthermia were further categorized into BT <38.6 degrees C or BT <39.2 degrees C, mean daily BtO2 was similar in all of the groups. When BT was 38.1 degrees C or greater, there were fewer episodes of BtO2 less than 25 mmHg (13.5 +/- 24.6; P < 0.05) and of BtO2 less than 15 mmHg (3.3 +/- 11.9; P < 0.05) than observed during brain normothermia. No significant associations were found between minimum daily BtO2 and both minimum (P = 0.81) and maximum (P = 0.19) daily BT or between maximum daily BtO2 and both minimum (P = 0.62) and maximum (P = 0.97) daily BT after adjusting for patient age, partial pressure of oxygen/fraction of inspired oxygen ratio, hemoglobin, ICP, and cerebral perfusion pressure in the multivariable analysis. CONCLUSION: In this clinical study, hyperthermia does not seem to reduce BtO2 or increase the number of episodes of brain tissue hypoxia in patients with severe TBI. These results suggest that hyperthermia may worsen outcome after TBI through mechanisms that may be separate from compromised brain oxygen. PMID- 18414136 TI - Tumor bed radiosurgery after resection of cerebral metastases. AB - OBJECTIVE: Adjuvant irradiation after resection of brain metastases reduces the risk of local recurrence. Whole-brain radiation therapy can be associated with significant neurotoxicity in long-term survivors of brain metastases. This retrospective study evaluates the role of tumor bed stereotactic radiosurgery as an alternative method of irradiation after initial resection of brain metastases to prevent local recurrence. METHODS: Forty patients underwent tumor bed radiosurgery after resection of brain metastases at two separate academic medical centers. The median age was 59.5 years. Twenty patients (67.5%) had single metastases. Resection was complete in 80% and partial in 20% of the patients. At the time of radiosurgery, systemic disease was active in 57.5%, inactive in 32.5%, and in remission in 10% of the patients. The median Karnofsky Performance Scale score was 80% (range, 60-100%). Radiosurgery was performed a median of 4 weeks after tumor resection. The median cavity radiosurgery volume was 9.1 ml (range, 0.6-39.9 ml). The median margin and maximum radiation dose were 16 and 32 Gy, respectively. RESULTS: Local control at the resection site was achieved in 73% of patients at a median follow-up period of 13 months. No variable significantly affected local control. New remote brain metastases occurred in 54% of the patients. Symptomatic radiation effect was seen in 5.4% of the patients. The median survival was 13 months after radiosurgery (range, 2-56 mo). CONCLUSION: Tumor bed radiosurgery provides effective local control of the tumor after resection in most patients. These preliminary data support radiosurgery after resection rather than traditional radiation therapy. PMID- 18414137 TI - Significant prevalence and genetic diversity of norovirus infection in Irish children. AB - Pediatric gastroenteritis places a considerable disease burden on children of the developed world. The national surveillance of gastroenteritis in Ireland is a combined virological and epidemiologic surveillance program. The objectives of this study were to characterize the norovirus (NoV) genotypes associated with viral gastroenteritis in children or=4 mo of age and determined that NoV and adenovirus infection are equally significant in children in the first 5 y of life. This group of pediatric patients reported diarrhea as their most common symptom raising the question whether Kaplan criteria are the most effective method for clinically diagnosing outbreaks of enteric infection in pediatric patients. ABBREVIATIONS: : PMID- 18414138 TI - The impact of constipation on growth in children. AB - The observation on the impact of constipation on nutritional and growth status in healthy children was never reported. During a 4-y period, we evaluated the consequence of constipation on growth in children. The enrolled children were aged between 1 and 15 y with constipation. Medical response of constipation to treatment was evaluated by the scoring of constipation symptoms. The correlation of therapeutic effect of constipation with growth status at 12 wk and 24 wk was statistically evaluated. About 2426 children (1284 boys, 1142 girls) with a mean age of 7.31 +/- 3.65 (range 1.1-14.9) y were enrolled. After 12-wk treatment, significant increase of z-scores of height-for-age, weight-for-age, and body mass index-for-age were all found in patients with good medical responses (1377 cases) than in those with poor medical responses (1049 cases). The 1049 patients with poor medical response received advanced medications; significant increase of z scores of height-for-age, weight-for-age, and body mass index were also found in these patients. A marked increase of appetite was significantly correlated with better gain on height and weight after treatment. We conclude that chronic constipation may retard growth status in children, and a long-term medication for constipation in children appears beneficial to their growth status. PMID- 18414139 TI - B-type natriuretic peptide to predict ductus intervention in infants <28 weeks. AB - Patent ductus arteriosus (PDA) is frequent in neonates with gestational age of less than 28 wk. Clinical and echocardiographic signs define hemodynamic significance of PDA, but do not reveal the need for PDA intervention in the first days of life. B-type natriuretic peptide (BNP) has been proposed as a screening tool for PDA in preterm infants. To determine whether BNP can predict the need for PDA intervention, plasma BNP was measured by chemiluminescence immunoassay in 67 preterm infants <28 wk (median 26) on the second day of life in a prospective blinded study. PDA intervention was based on specified clinical and echocardiographic findings. Twenty-four patients (intervention group) received treatment for PDA and 43 patients (controls) remained without intervention. BNP concentrations were higher in the intervention (median 1069 pg/mL) than in the control group (247 pg/mL, p < 0.001). BNP correlated positively with ductal size (R = 0.46, p < 0.001) and atrial/aortic root ratio (R = 0.54, p < 0.001). In conclusion, plasma BNP proved to be a good predictor for ductus intervention (area under the curve: 0.86) with the best cutoff at 550 pg/mL on the second day of life in ventilated infants less than 28 wk gestation (sensitivity: 83%; specificity: 86%). PMID- 18414140 TI - Novel red cell indices indicating reduced availability of iron are associated with high erythropoietin concentration and low ph level in the venous cord blood of newborns. AB - There is evidence that an elevated erythropoietin (EPO) concentration is associated with signs of iron deficient erythropoiesis. The aim of this study was to evaluate the iron status by means of novel cellular indices and serum iron markers and to determine whether these are associated with EPO and pH in the venous cord blood of 193 full-term newborns. There were positive correlations between EPO and the percentage of hypochromic red blood cells (%HYPOm) and reticulocytes (%HYPOr) [r = 0.45 (p < 0.001) and r = 0.56 (p < 0.001), respectively]. %HYPOm and %HYPOr also had negative correlations with pH [r = 0.53 (p = 0.001) and r = -0.46 (p = 0.001), respectively]. Newborns who had low pH (pH < or =7.15, n = 16) had significantly higher %HYPOm, %HYPOr, and serum transferrin receptor and transferrin concentrations in their cord blood than newborns with normal pH. Thus, in newborn cord blood, the higher number of red cells and reticulocytes with lower Hb content may have impaired the oxygen carrying capacity that has been a trigger for EPO production. Furthermore, signs of lower hemoglobinization of red cells are associated with low umbilical vein pH in the newborns, indicating an increased risk of birth asphyxia. PMID- 18414141 TI - Model mice for mild-form glycine encephalopathy: behavioral and biochemical characterizations and efficacy of antagonists for the glycine binding site of N methyl D-aspartate receptor. AB - Glycine encephalopathy (GE) is caused by an inherited deficiency of the glycine cleavage system (GCS) and characterized by accumulation of glycine in body fluids and various neurologic symptoms. Coma and convulsions develop in neonates in typical GE while psychomotor retardation and behavioral abnormalities in infancy and childhood are observed in mild GE. Recently, we have established a transgenic mouse line (low-GCS) with reduced GCS activity (29% of wild-type (WT) C57BL/6) and accumulation of glycine in the brain (Stroke, 2007; 38:2157). The purpose of the present study is to characterize behavioral features of the low-GCS mouse as a model of mild GE. Two other transgenic mouse lines were also analyzed: high-GCS mice with elevated GCS activity and low-GCS-2 mice with reduced GCS activity. As compared with controls, low-GCS mice manifested increased seizure susceptibility, aggressiveness and anxiety-like activity, which resembled abnormal behaviors reported in mild GE, whereas high-GCS mice were less sensitive to seizures, hypoactive and less anxious. Antagonists for the glycine-binding site of the N methyl-D-aspartate receptor significantly ameliorated elevated locomotor activity and seizure susceptibility in the low-GCS mice. Our results suggest the usefulness of low-GCS mice as a mouse model for mild GE and a novel therapeutic strategy. PMID- 18414142 TI - Safety experience with bosentan in 146 children 2-11 years old with pulmonary arterial hypertension: results from the European Postmarketing Surveillance program. AB - The oral dual endothelin receptor antagonist bosentan has been shown to improve the short- and medium-term course of adult pulmonary arterial hypertension (PAH); however, data from clinical studies in children are limited. This analysis investigated the safety profile of bosentan in pediatric patients in a European, prospective, noninterventional, Internet-based postmarketing surveillance database (Tracleer PMS). Pediatric patients (aged 2-11 y) were compared with patients aged > or =12 y. Over a 30-mo period, 4994 patients, including 146 bosentan-naive pediatric patients (51.4% males), were captured in the database. Predominant etiologies in children were idiopathic PAH (40.4%) and PAH related to congenital heart disease (45.2%). The majority of children were in New York Heart Association functional class II (28.1%) or III (50.7%), and median exposure to bosentan was 29.1 wk. Elevated aminotransferases were reported in 2.7% of children versus 7.8% of patients > or =12 y. The discontinuation rate was 14.4% in children versus 28.1% in patients > or =12 y. The Tracleer PMS results provide unique information on pediatric PAH in Europe. They also suggest that Tracleer may be better tolerated in children than in adults. This observation confirms the value of monthly monitoring of liver function for the duration of bosentan treatment. PMID- 18414143 TI - Prenatal smoking exposure and neonatal DNA damage in relation to birth outcomes. AB - This study investigated whether mothers with prenatal environmental tobacco smoke (ETS) exposure increased the newborn genetic damage and adverse birth outcomes. Study participants were women receiving prenatal care at three hospitals in Central Taiwan and their newborns. Participants were divided into two groups (nonsmokers and ETS-exposed non-smokers) based on maternal ETS-exposed status. Comet assay were performed for cord blood samples. Infants born to mothers with prenatal ETS exposure had the highest mean cord blood DNA damage score (69.7 +/- 42.3) and poorer birth outcomes. No negative fetal growth effects appeared among newborns with low DNA damage levels. Among newborns with high DNA damage levels (comet scores >50), those born to prenatal ETS exposure had an average reduction of 252.7 g in birth weight, 1.10 cm shorter in length and a 0.92-cm decrease in head circumference, compared to newborns with no smoking exposure. This study shows that the DNA damage scores can be used as an effect-modifier on the relationships between ETS exposure and adverse birth outcome. The association appears more apparent for the ETS exposure in relation with more severe DNA damage. PMID- 18414144 TI - Relationship between functional residual capacity and oxygen desaturation during short central apneic events during sleep in "late preterm" infants. AB - Apneic episodes are frequent in the preterm neonate and particularly in active sleep (AS), when functional residual capacity (FRC) can be decreased. Furthermore, FRC may be inversely correlated with the speed of blood-O(2) desaturation. We evaluated the potential involvement of FRC in the mechanisms responsible for blood-O(2)-desaturation during short central apneic events (>3 s) in "late-preterm" infants and analyzed the specific influence of sleep state. Apneic events were scored in 29 neonates (postmenstrual age: 36.1 +/- 1.2 wk) during AS and quiet sleep (QS). FRC was measured during well-established periods of regular breathing. Apneas with blood-O(2)-desaturation (drop in SpO(2) >5% from the baseline, lowest SpO(2) during apnea: 91.4 +/- 1.8%) were more frequent in AS than in QS, whereas no difference was seen for apneas without desaturation. The magnitude of the FRC did not depend on the sleep state. In AS only, there was a negative relationship between FRC and the proportion of apneas with desaturation. Even in late preterm infants who do not experience long-lasting apnea, blood-O(2)-desaturation during short apneic events is related (in AS but not QS) to a low baseline FRC. Sleep stage differences argue for a major role of AS-related mechanisms in the occurrence of these apneas. PMID- 18414146 TI - 2007 E. Mead Johnson award: scientific pursuit of the allergy problem. AB - My research has focused on elucidating the allergy problem over the past two decades. The primary approach has been to uncover critical mechanisms of allergic inflammation, with particular focus on eosinophils, a hallmark cellular constituent of allergic responses. Molecular processes that bridge T helper cell type 2 (TH2) immunity with eosinophilia and key checkpoints for regulating eosinophilia have been uncovered. Notably, interleukin (IL)-5 (derived from TH2 cells) has been identified as the chief hematopoietin responsible for eosinophil expansion in the circulation. Pathways for selective eosinophil mobilization from the blood stream to the tissue have been uncovered by defining the role of the eotaxin subfamily of chemokines in eosinophil chemoattraction and activation. Finally, TH2 cell derived IL-4 and IL-13 have been defined as chief inducers of the eotaxins, and upstream orchestrators of eosinophilic inflammation. These translational studies have formulated novel therapeutic strategies (currently being tested) for a variety of eosinophilic conditions, with particular attention on hypereosinophilic syndromes and eosinophil-associated gastrointestinal disorders such as eosinophilic esophagitis. PMID- 18414145 TI - N-carbamylglutamate markedly enhances ureagenesis in N-acetylglutamate deficiency and propionic acidemia as measured by isotopic incorporation and blood biomarkers. AB - N-acetylglutamate (NAG) is an endogenous essential cofactor for conversion of ammonia to urea in the liver. Deficiency of NAG causes hyperammonemia and occurs because of inherited deficiency of its producing enzyme, NAG synthase (NAGS), or interference with its function by short fatty acid derivatives. N carbamylglutamate (NCG) can ameliorate hyperammonemia from NAGS deficiency and propionic and methylmalonic acidemia. We developed a stable isotope (13)C tracer method to measure ureagenesis and to evaluate the effect of NCG in humans. Seventeen healthy adults were investigated for the incorporation of (13)C label into urea. [(13)C]urea appeared in the blood within minutes, reaching maximum by 100 min, whereas breath (13)CO(2) reached a maximum by 60 min. A patient with NAGS deficiency showed very little urea labeling before treatment with NCG and normal labeling thereafter. Correspondingly, plasma levels of ammonia and glutamine decreased markedly and urea tripled after NCG treatment. Similarly, in a patient with propionic acidemia, NCG treatment resulted in a marked increase in urea labeling and decrease in glutamine, alanine, and glycine. These results provide a reliable method for measuring the effect of NCG on nitrogen metabolism and strongly suggest that NCG could be an effective treatment for inherited and secondary NAGS deficiency. PMID- 18414148 TI - The significance of N-cadherin expression in breast cancers: an evolving story. PMID- 18414149 TI - Patient's page. Malignant melanoma and the importance of screening. PMID- 18414150 TI - Cardiac tamponade: an unusual complication of simultaneous treatment of tuberculosis and HIV. AB - Human immunodeficiency virus (HIV) has caused resurgence in tuberculosis (TB) worldwide. HIV-TB coinfected individuals are at increased risk for complications of TB and HIV treatment, such as adverse drug reactions and immune reconstitution syndrome. A 17-year-old male with HIV-TB coinfection, who developed cardiac tamponade from immune reconstitution during treatment for TB and HIV, is reported in this document. PMID- 18414151 TI - A case of black magic. AB - The authors present the case of a 64-year-old woman who presented with massive subcutaneous emphysema 2 weeks after unknowingly inhaling a Brazil nut. Foreign body inhalation should be a differential diagnosis in all such patients, with computed tomography scanning of the chest being the most appropriate investigation. PMID- 18414152 TI - Multidrug-resistant viridans streptococcus (MDRVS) osteomyelitis of the mandible successfully treated with moxifloxacin. AB - Multidrug-resistant viridans group streptococcus (MDRVS) strains have emerged as important pathogens. Treatment of MDRVS infections is problematic. The use of fluoroquinolones for treatment of MDRVS osteomyelitis has not been established. We present the first case of MDRVS osteomyelitis of the mandible successfully treated with sequential intravenous then oral moxifloxacin, and review the literature on the subject. PMID- 18414155 TI - Respiratory CSI: diagnosing occupationally related lung disease. PMID- 18414153 TI - Comparison of neopterin levels in patients with coronary artery ectasia versus patients with obstructive coronary artery disease. AB - OBJECTIVES: Inflammation has been reported as an important component of vascular aneurysm formation, as found in obstructive vascular disorder. Neopterin is produced by activated macrophages and is thought to represent a marker of immune activation and macrophage activity. The plasma neopterin levels were investigated in coronary artery ectasia (CAE) patients to investigate the role of inflammation. STUDY DESIGN: The study population consisted of three groups: the first consisted of 28 patients with isolated CAE without stenotic lesion; the second of 27 patients with obstructive coronary artery disease (O-CAD) without CAE; and the third group of 15 control subjects with normal coronary arteries (NCA). Plasma soluble neopterin levels were measured in all patients and control subjects using commercially available enzyme-linked immunosorbent assay kits. RESULTS: Plasma neopterin levels were found to be significantly higher in patients with isolated CAE compared with control subjects with angiographically NCA (18.5 +/- 8.8 versus 8.7 +/- 2.6 nmol/L, respectively, P = 0.006). Although neopterin levels were higher in patients with CAE than in patients with O-CAD, they did not reach the statistically significant levels (18.5 +/- 8.8 versus 16.8 +/- 8.2 nmol/L, respectively, P = 0.77). Patients with O-CAD had significantly higher levels of neopterin compared with subjects with angiographically NCA (16.8 +/- 8.2 versus 8.7 +/- 2.6 nmol/L, respectively, P = 0.03). The mean serum neopterin levels in patients with single-vessel, two-vessel, and diffuse ectasia were as follows: 17.4 +/- 9.9 nmol/L, 19.5 +/- 8.9 nmol/L, and 20 +/- 5.5 nmol/L, respectively (P = 0.4). CONCLUSION: Patients with isolated CAE have raised levels of neopterin compared with patients with NCA, showing the possible role of inflammatory processes (monocyte/macrophage activity) in the higher levels of neopterin in patients with O-CAD. PMID- 18414156 TI - Neopterin: its role in inflammation and coronary heart disease. PMID- 18414157 TI - A new stigmata. PMID- 18414158 TI - A case of flavor-induced lung disease. AB - Bronchiolitis obliterans, a rare inflammatory disorder of the small airways, has been associated with various infections, aspiration, toxic fume inhalation, drugs, transplants, and collagen vascular disorders. In May 2000, this condition was described in eight workers in a microwave popcorn plant in Missouri. Subsequent studies reported its association with exposure to volatile organic compounds used in butter flavoring. Diacetyl was found to be the most predominant of these compounds. We report a case of this rare but serious disease in a 41 year-old flavor industry worker who was initially diagnosed with occupational asthma. PMID- 18414159 TI - Can medicine be compared with art? PMID- 18414160 TI - A case of paraneoplastic cerebellar ataxia secondary to renal cell carcinoma. AB - A 64-year-old female patient presented with acute cerebellar ataxia. After ruling out vascular, infectious, metastatic, demyelinating, and medication etiologies, a paraneoplastic origin for her illness was determined. A renal mass was discovered and found to be renal cell carcinoma. Her symptoms resolved after a radical nephrectomy. Physicians should consider the diagnosis of paraneoplastic neurologic syndromes whenever a thorough evaluation fails to explain neurologic abnormalities. PMID- 18414161 TI - Human trafficking and the healthcare professional. AB - Despite the legislation passed in the 19th century outlawing human slavery, it is more widespread today than at the conclusion of the civil war. Modern human slavery, termed human trafficking, comes in several forms. The most common type of human trafficking is sex trafficking, the sale of women and children into prostitution. Labor trafficking is the sale of men, women, and children into hard labor for which they receive little or no compensation. Other forms of trafficking include child soldiering, war brides, and organ removal. Healthcare professionals play a critical role in both finding victims of human trafficking while they are still in captivity, as well as caring for their mental and physical needs upon release. Those working in the healthcare profession need to be educated regarding how a trafficking victim may present, as well as their unique healthcare needs. PMID- 18414162 TI - Hyperammonemic coma: beyond hepatic encephalopathy. PMID- 18414163 TI - A 30-month experience of thoracic empyema in a tertiary hospital: emphasis on differing bacteriology and outcome between the medical intensive care unit (MICU) and medical ward. AB - OBJECTIVES: To analyze the causative pathogens and outcomes of patients with thoracic empyema admitted to the medical intensive care unit (MICU) and medical ward. METHODS: We prospectively studied the empyemic patients in the MICU and retrospectively analyzed the medical records of empyemic patients in the medical ward treated in a tertiary university hospital from April 2001 to September 2003. RESULTS: During this period, 116 patients in the medical ward and 78 patients in MICU had complicated parapneumonic effusions or empyemas. Effusion cultures were positive in 164 patients (85%); a total of 147 and 78 microorganisms were isolated from the 106 medical ward patients and 58 MICU patients, respectively. No matter whether medical ward or MICU patients, aerobic gram-negative organisms were the most common bacteria in positive-culture effusions (110, 67%). Klebsiella pneumoniae (14, 24%) was the predominant pathogen among the MICU patients, and Streptococcus spp. (28, 26%) was the main pathogen among the medical ward patients. Compared with these positive-culture empyemic patients in the medical ward, MICU patients had a significantly higher percentage of aerobic gram-negative organism infections (P = 0.034) and a higher infection-related mortality rate (P = 0.01). CONCLUSION: The mortality and predominant pathogens in patients with complicated parapneumonic effusions or thoracic empyemas in the medical ward and MICU were different. The increasing gram-negative pathogens in empyemas have become an urgent problem. PMID- 18414164 TI - Change in residents' perceptions of teaching: following a one day "Residents as Teachers" (RasT) workshop. AB - OBJECTIVES: The objective of this study was to assess the perceptions and attitudes of resident physicians toward teaching before and after participation in a mandatory "Residents as Teachers" (RasT) workshop in four domains: (1) setting goals and expectations, (2) use of clinical microskills in teaching, (3) evaluation and feedback, and (4) enthusiasm and preparedness toward teaching. METHODS: Pre- and postintervention questionnaires were utilized. Data were analyzed for all respondents. Subgroup analyses were performed for each academic year and for primary care versus nonprimary care specialties. RESULTS: Over a 5 year period, 15 RasT workshops were presented to 276 residents from 10 different residency programs. Eighty-six percent completed the questionnaire before participation in the workshop, and 88% completed the questionnaire immediately after participation. The difference between the mean post-RasT and pre-RasT ratings on each item was used to measure the change in that item resulting from participation in the workshop. CONCLUSION: Overall, residents' self-assessed ratings of their attitudes toward teaching were positively impacted by participation in a RasT workshop. Further subanalysis showed that residents in primary care specialties showed a significantly greater increase in their ratings than residents in nonprimary care specialties. PMID- 18414165 TI - A primary tracheal carcinoid tumor masquerading as chronic obstructive pulmonary disease. AB - Primary tumors of the trachea are rare and can be missed because of the paucity of symptoms and findings and the difficulty in detecting them with chest radiographs. A 31-year-old male patient was admitted with complaints of shortness of breath, coughing, phlegm, and blood in the sputum. He stated that he was being treated for chronic obstructive pulmonary disease. Fiberoptic bronchoscopy revealed a vegetative mass with a wide base on the posterolateral wall of the distal one-third of the trachea. Postoperative histopathological examination revealed a typical carcinoid tumor. In patients with an unexplained cough, dyspnea, infrequent hemoptysis, and normal pulmonary imaging findings, tracheal carcinoma should be suspected. In such cases, early thoracic computed tomography and bronchoscopic examination can provide a rapid diagnosis and treatment options and prevent a false diagnosis. PMID- 18414166 TI - Necrotizing panniculitis: a skin condition associated with acinar cell carcinoma of the pancreas. AB - Pancreatic panniculitis (PP) is a rare cutaneous eruption that is associated with severe pancreatic disease. A patient presented with a fever, joint pains, and an erythematous rash with draining pustules that had spread from his legs to his arms over 4 months. Thorough investigation revealed stage IV acinar cell carcinoma of the pancreas. The rash was a result of necrotizing PP. The variable cutaneous manifestations of internal malignancies may challenge primary care physicians and dermatologists when patients present without findings associated with malignancy. Panniculitis should be kept in mind in the differential diagnosis of inflamed appearing nodules and pustules with an erythematous base, particularly when they are progressive and unrelenting. PMID- 18414167 TI - Postchemotherapy hyperammonemic encephalopathy emulating ornithine transcarbamoylase (OTC) deficiency. AB - A young patient with hepatocellular carcinoma receiving chemotherapy presented with encephalopathy. Evaluation of the patient revealed a metabolic profile consistent with ornithine transcarbamoylase (OTC) deficiency, an inherited disorder of the urea cycle. The evaluation yielded a plasma amino acid analysis consistent with OTC deficiency. However, genetic analysis did not reveal a somatic mutation of the OTC gene in this patient. The hyperammonemic encephalopathy was reversed by the infusion of arginine, a common treatment for hereditary OTC deficiency. This case may represent a distinct syndrome of reversible hyperammonemia in patients with hepatocellular carcinoma. PMID- 18414169 TI - Burkitt lymphoma presenting with hypercalcemic emergency. PMID- 18414170 TI - C-reactive protein: the harbinger of cardiovascular diseases. AB - Coronary heart disease remains the major cause of mortality and morbidity in the United States and other western countries despite major advances in its treatment. During the last decades, many markers of coronary artery disease have been found which help predict future risk of cardiovascular events. High sensitivity C-reactive protein has been studied extensively and was seen to be associated with a higher risk of cardiovascular events in patients with acute coronary syndromes and/or peripheral artery disease. Discussed in this review is the latest literature about this marker and its association with cardiovascular disease, as well as the latest therapeutic options available. PMID- 18414171 TI - Endobronchial ultrasound in the evaluation of lung cancer: a practical review and cost analysis for the practicing pulmonologist. AB - Flexible bronchoscopy remains an important tool in the staging, diagnosis, and treatment of primary and metastatic lung malignancies. Endobronchial ultrasound is a new technology utilized with bronchoscopy that has been shown to identify bronchial wall invasion by malignant tumors, aid in the fine needle aspiration of peripheral lung lesions and mediastinal/hilar lymph nodes, and determine the course of treatment in patients with pulmonary carcinoma in situ. The decision to invest both time and money in this technology is determined by several factors such as the cost of the equipment, reimbursement for the procedure, availability of training, the number of bronchoscopies one performs in a year, and access to endoscopic ultrasound and mediastinoscopy. This article reviews the literature to determine the utility of endobronchial ultrasound in the management of patients with lung cancer and to provide information to practicing pulmonologists that may aid in determining whether and where this technology fits into their clinical armamentarium. PMID- 18414172 TI - Colchicine exposures: the Texas experience. AB - OBJECTIVES: Colchicine is a relatively uncommon toxin, but is known to precipitate severe multiorgan failure in overdose. Little is known about exposure patterns and outcomes in cases of colchicine ingestion. Our goal was to add to toxicologic knowledge through a database review and descriptive study of colchicine exposures. METHODS: Texas Poison Center Network Data was reviewed for the years 2000 to 2005, and all reports of colchicine exposures were reviewed. RESULTS: A total of 79 cases were found in the time period studied. The most common exposure reasons were unintentional-therapeutic error (33%), unintentional general (28%), and intentional-suspected suicide (18%). Medical outcomes included no effect (24%), minor effect (20%), moderate effect (15%), and major effect (3%). The most common clinical findings included vomiting (20%), diarrhea (17%), and abdominal pain (7%). The most commonly employed therapies were dilution (28%), single-dose activated charcoal (26%), cathartics (16%), and gastrointestinal lavage (15%). CONCLUSION: The majority of cases of exposure produced no significant effects, and fatality was uncommon in this sample. Colchicine is a relatively uncommon toxin among therapeutic drugs, and though capable of it, is rarely responsible for significant morbidity or mortality. Meticulous exposure record keeping at poison centers is a key to the study of patterns of toxicity with uncommon toxins such as colchicine. PMID- 18414173 TI - Poor outcomes associated with drainage of pericardial effusions in patients with pulmonary arterial hypertension. AB - OBJECTIVES: Pulmonary arterial hypertension (PAH) in its advanced stages is complicated by right heart failure and often pericardial effusion. The optimal treatment of large or hemodynamically significant pericardial effusions in this group has not been defined. METHODS: All patients followed at the Johns Hopkins Hospital for PAH during a 1-year period that underwent pericardiocentesis or pericardial window placement were identified. Charts were analyzed for patient characteristics, echocardiographic data, and type/outcome of procedure. RESULTS: Six patients were identified; five underwent therapeutic drainage. Pericardiocentesis was performed in four cases; two had surgical pericardial windows. Two patients died after pericardiocentesis and one patient died after surgery. All patients died within 13 hours of the procedure. CONCLUSION: We found a high mortality related to pericardial fluid drainage in patients with PAH. The pathophysiologic explanation for these deaths remains unclear, but clinicians should consider conservative management in this situation if possible. PMID- 18414174 TI - Vascular access infection among hemodialysis patients in Northern Jordan: incidence and risk factors. AB - OBJECTIVES: Vascular access infection (VAI) causes significant mortality and morbidity in hemodialysis patients. The aim of this prospective, descriptive, exploratory survey of end-stage renal disease patients was to explore the incidence and risk factors of VAI. METHODS: A total of 188 patients were included in the study from five different hospital/dialysis units in northern Jordan. All patients answered a questionnaire related to their vascular access (VA) and infection. Demographic information, etiology of renal failure, and infection history related to the access were also collected and analyzed. RESULTS: Diabetes mellitus was the most common etiology of renal failure, followed by hypertension, urinary tract infection, and congenital malformation. The infection rate was variable according to the hospital and the type of VA; of 188 patients, 65 patients had a VAI during the year of study. Of these, 36 of 64 were because of catheters, 24 of 105 were because of arteriovenous fistulas (AVFs), and 5 of 19 were because of arteriovenous graft. Of the 65 with a VAI, 58 patients had a low level of education. CONCLUSIONS: VAI was common in hemodialysis patients, and the risk varied substantially among different VA types and different dialysis units. The infection rate from catheters and AVFs was similar to that reported in many studies, but the infection rate in the arteriovenous graft group was lower than that for AVFs. These results can be improved by strictly following the National Kidney Foundation Disease Output Quality Initiative clinical practice guidelines for dialysis access. PMID- 18414175 TI - The Kentucky SimSmoke tobacco policy simulation model: reaching Healthy People 2010 goals through policy change. AB - OBJECTIVES: Healthy People (HP) 2010 has a goal of reducing adult smoking prevalence to 12% by 2010. Kentucky, with the highest adult smoking rates (28.7% versus a US average of 20.9%) and weak tobacco control policies, has the potential to reduce smoking by implementing stricter policies. METHODS: The SimSmoke tobacco control policy model is applied to Kentucky, validated over 1993 to 2003. Smoking prevalence evolves over time through initiation and cessation, as behaviors influenced by tobacco control policies. RESULTS: SimSmoke predicts well over the pre-2007 period, including larger annual decreases in latter years. With policies suggested by HP 2010 goals, smoking prevalence is projected to fall to about 19% by 2011 and 14% by 2026; more than 17,000 smoking-attributable deaths would be avoided by 2026. CONCLUSIONS: SimSmoke suggests policies can have a large impact on smoking rates and save lives in Kentucky. If HP 2010 goals will be met, states such as Kentucky need to implement strong tobacco control policies. PMID- 18414176 TI - Is N-cadherin expression important in ductal carcinoma? AB - OBJECTIVES: Neural (N)-cadherin is a calcium-dependent cell adhesion molecule that is associated with invasive tumors in breast cancer, but no association with grade or nodal status has been shown in previous studies. The present study examined the expression of N-cadherin in human breast cancer and assessed its prognostic value in long-term patient follow-up (10 years). METHODS: Using real time polymerase chain reaction, the number of N-cadherin transcripts in normal breasts (n = 32) and infiltrating ductal carcinomas (n = 90) was assessed. The results were then analyzed in relation to grade, nodal involvement, distant metastasis, TNM stage, Nottingham Prognostic Index, and survival over 10 years. RESULTS: The levels of N-cadherin transcripts (normalized to glyceraldehyde 3 phosphate dehydrogenase) in primary tumors were lower in patients with metastases (P = 0.05), with local recurrence (P = 0.037), and those who died as a result of breast cancer (P = 0.038). There was no significant association with grade, nodal status, TNM stage, or Nottingham Prognostic Index. CONCLUSION: In ductal carcinomas, decreased levels of N-cadherin (normalized to glyceraldehyde 3 phosphate dehydrogenase) in primary tumors correlate with local recurrence and death in long-term follow-up of patients. PMID- 18414177 TI - Osteomyelitis of the mandible. PMID- 18414178 TI - Diagnosis of marginal cell lymphoma of small intestine by double balloon enteroscopy. AB - A 65-year-old woman with a medical history of diabetes mellitus type 2, hypertension, an old cerebrovascular accident, and seizure disorder presented to the emergency room with lower abdominal pain of 4 weeks duration. Upon physical examination, her abdomen was soft and bowel sounds were present, but there was diffuse tenderness in her lower abdomen with some guarding. A computed tomography scan of her abdomen with oral and intravenous contrast showed significantly thickened small bowel loops with subjacent lymphadenopathy. Biopsies obtained during esophagogastroduodenoscopy and colonoscopy showed acute and chronic inflammation. A double balloon enteroscopy (DBE) was then performed, which showed stricture in the jejunum from which the biopsy was obtained. The biopsy showed marginal cell lymphoma. The patient is presently undergoing chemotherapy. Double balloon enteroscopy is a new elegant endoscopical technique that seems promising, as the endoscopist can reach undiscovered small bowel segments. It seems to be well tolerated and safe. For the first time, it provides the means to endoscopically investigate and treat disorders of the small intestine that have previously been inaccessible to conventional endoscopy. PMID- 18414179 TI - Patient gender affects skin cancer screening practices and attitudes among veterans. AB - BACKGROUND: Skin cancer screening (SCS) with a full body skin examination (FBSE) has the potential to reduce morbidity and mortality. Little is known about gender differences with respect to SCS practices and attitudes between men and women. METHODS: Data from two previously published studies based on questionnaires administered to veterans were combined and analyzed according to patient gender. The participants consisted of a convenience sample of 437 patients awaiting primary care, women's health, or dermatology clinic appointments at the West Haven Veterans Affairs Medical Center. RESULTS: Male veterans were more likely to report undergoing FBSE than female veterans (32 versus 18%), but less likely to perform self-examination (42 versus 48%). Female veterans were more likely to report embarrassment than men, but both genders expressed that providers who perform SCS are thorough. Gender discordance between patient and examining physician is more likely to lead to refusal for women than men (16 versus 2%). CONCLUSION: We found low rates of SCS in both male and female veterans in different clinic settings at a Veterans Affairs Medical Center. Female veterans are less likely to report undergoing FBSE and more likely than male veterans to perform self-examination for skin cancer, to report embarrassment with FBSE, and refuse FBSE if the examining physician is of the opposite gender. PMID- 18414180 TI - Psychotic mania associated with mefloquine in a bipolar patient. AB - A 63-year-old male patient with a history of bipolar I disorder presented to the emergency department in an acutely psychotic state. The patient had just returned from vacation in a malarial high-risk area and, as a result, had taken five weekly doses of mefloquine for prophylaxis. The patient's bipolar disorder was being treated with lithium, and he had been stable for about 8 years. All organic causes of psychosis were ruled out. The patient was admitted to the inpatient psychiatric unit and was treated with quetiapine until he was stable enough to return home. PMID- 18414181 TI - Flavanol-rich cocoa: a cardioprotective nutraceutical. AB - A growing number of epidemiologic studies have shown a link between the ingestion of flavonoid-rich cocoa and cardiovascular health benefits. A particular kind of flavonoid found in cocoa, namely flavan-3-ol (flavanol), has recently received more attention in this context. This review discusses the cardioprotective properties of flavanols, and the mechanisms underlying these beneficial actions. PMID- 18414182 TI - Coronary artery aneurysms. AB - Aneurysmal coronary artery disease is frequently encountered in clinical cardiology practice. Although more commonly associated with atherosclerosis, a variety of other acquired (eg, inflammatory, infectious, iatrogenic) or congenital causes have been identified. Recent research on the pathogenesis of coronary aneurysms has yielded interesting results. Advances in imaging have also provided new insights as to the nature of angiographic coronary aneurysms. Critical assessment of the abnormal flow dynamics and pathophysiology of aneurysms has been performed and there is an improved understanding of the associated complications. We present an extensive review of the recent literature highlighting the major advances in the field. PMID- 18414183 TI - Does body mass index really matter in the management of heart failure?: a review of the literature. AB - Excess body weight increases the risk for many disorders including cardiovascular disease and such patients have a greater risk for developing heart failure (HF). Despite evidence demonstrating the adverse effects of excess weight, the relationship between body mass index (BMI) and mortality in HF patients remains controversial. Paradoxically, several large cohort studies have shown that overweight and obese HF patients seem to have better survival than their healthy weight counterparts. The exact mechanism for this "obesity paradox" is not fully understood. Proposed mechanisms include a greater tolerance to angiotensin converting enzyme inhibition, higher serum lipid levels, and the alteration of inflammatory cytokine metabolism in obese patients. Although the relationship between elevated BMI and improved survival has been well documented, recent clinical trials have not addressed this association. In 65 of 75 clinical HF trials reviewed, BMI as a potential independent predictor of outcomes was not addressed. Furthermore, the variation of pharmacokinetics in the obese population has been dealt with to a limited degree. If data concerning BMI and weight loss is to directly impact treatment recommendations for HF patients, well-designed clinical trials are needed. PMID- 18414184 TI - Fibrate therapy: an update. AB - Fibrates are a class of lipid-lowering medication primarily used as second-line agents behind statins. Acting via the peroxisome proliferators-activated receptor alpha, their main lipoprotein effects are to lower serum triglyceride levels and to raise high-density lipoprotein-cholesterol, with modest effects on low-density lipoprotein-cholesterol. However, many clinical trials indicate that fibrates may have benefits beyond simply altering one's lipid profile. Several angiographic studies show retardation in the progression of atherosclerotic lesions in coronary vessels. Although clinical trials have failed to show a reduction in mortality with fibrates, several post hoc analyses indicate that there may be a mortality benefit in patients with features of the metabolic syndrome. Given that fibrates are often used as second-line agents, it is essential they are safe to be given in combination with other agents, particularly statins and ezetimibe. Although the side-effect profile of fibrates includes gastrointestinal symptoms, increased liver function tests, a reversible rise in creatinine and myositis, in general, fibrates seem to be safe to use in combination with other lipid lowering medications. Thus far, fibrates have not shown a mortality benefit in randomized clinical trials; as a result, they cannot be considered first-line medication for the primary or secondary prevention of coronary artery disease. PMID- 18414185 TI - Urotensin II: a new pharmacologic target in the treatment of cardiovascular disease. AB - Urotensin II was first identified over 30 years ago as a potent vasoconstrictor, and the identification of its receptor in the heart, lungs, blood vessels, and brain have made it a potential target for human pharmacotherapy. Current research would suggest that urotensin II plays a major role in the pathophysiology of various cardiovascular disease entities. This article discusses the biologic effects of urotensin under normal and pathophysiologic conditions, and reviews the research experiences with synthetic urotensin blockers in the treatment of various cardiovascular illnesses. PMID- 18414186 TI - HDL metabolism and CETP inhibition. AB - High density lipoprotein-cholesterol (HDL-C) concentration in the blood is independently and inversely associated with an increased risk of coronary heart disease. Some of the cholesterol-lowering drugs (niacin, fibrates, and statins) incidentally raise HDL-C. These drugs are not effective in causing major changes in HDL-C. Since the discovery of human genetic cholesteryl ester transfer protein (CETP) deficiency in a Japanese population with high levels of HDL-C and apolipoprotein A-I, CETP inhibition has become a novel strategy for raising HDL-C in humans. Mice, a species naturally lacking CETP, were transduced with the human CETP gene, which resulted in dose-related reductions in HDL-C. Rabbits, a species with naturally high levels of CETP, were fed a synthetic CETP inhibitor, JTT-705, leading to both a 90% increase in HDL-C and a 70% reduction in aortic atherosclerotic lesion area. Human intervention trials with a new potent and selective CETP inhibitor, torcetrapib, have taken place. In a phase I multidose trial, HDL-C increased by 91% with torcetrapib 120 mg twice daily. A phase II trial conducted with multiple combinations of torcetrapib and atorvastatin showed that the combination was well tolerated and doses 30 mg and higher of torcetrapib caused 8.3-40.2% changes from baseline HDL-C across the dose range of atorvastatin at 12 weeks. Recently the phase III clinical trial ILLUMINATE (Investigation of Lipid Level Management to Understand its Impact in Atherosclerotic Events) was prematurely terminated because of an increase in mortality in the torcetrapib/atorvastatin treatment arm compared with atorvastatin used alone. In companion studies no improvement in carotid or coronary atherosclerosis could be detected in patients treated with the torcetrapib/atorvastatin combination despite favorable changes in both low density lipoprotein (LDL)- and HDL-cholesterol levels. The future for CETP inhibition with drug therapy is now unclear, and must include a closer look at CETP inhibitor's effects on blood pressure and HDL itself. Accordingly, it was recently shown in 2 double-blind, placebo-controlled, randomized, phase I studies with the CETP inhibitor anacetrapib in healthy individuals and in patients with dyslipidemias that the drug increased HDL and reduced LDL, while having no effect on blood pressure. PMID- 18414187 TI - Differences in stage of disease between migrant women and native Danish women diagnosed with cancer: results from a population-based cohort study. AB - The aim of the study is to compare differences in cancer stage at diagnosis between migrant women and native Danish women. The stage is used as a clinical indicator of access to healthcare until the point of diagnosis. Refugees and family reunited migrants who received residence permits in Denmark from 1 January 1993 to 31 December 1999 were included and matched 1 : 4 on age and sex with a Danish-born reference population. Our final female population included 24 734 migrants and 123 670 controls. Civil registration numbers of the cohort were linked to the Danish Cancer Registry whereby cases were identified in the period 1.1.1993-31.12.2002. Only women from Eastern Europe and the Middle East were included. This amounted to 269 migrants and 1608 native Danes. Data from the Danish Cancer Registry included diagnosis, time of diagnosis and disease stage at diagnosis. Our initial analyses of migrant subgroups showed that migrant women had decreased odds ratios of being diagnosed at the local stage and increased odds of having unknown stage, although these tendencies were mainly not statistically significant. A subsequent analysis of an overall migrant effect on all cancer sites emphasized these tendencies. This analysis reached borderline significance for local versus nonlocal stage and significance for unknown versus known stage. Our results indicate that migrant women may experience barriers in access to healthcare until cancer diagnosis compared with Danish women. More research is, however, needed to confirm our results and to find out if they indicate general problems concerning migrants' access to healthcare in Denmark. PMID- 18414188 TI - European studies on long-term exposure to ambient particulate matter and lung cancer. AB - European epidemiological studies on ambient air pollution and cancer published before December 2006 are reviewed, with focus on five analytic studies providing data on the association between various measures of particulate matter (PM) and lung cancer. A case-control study of 755 men who died from lung cancer in Trieste, Italy, reported that, compared with less than 0.18 g/m/day of deposition of particulate, the relative risk (RR) was 1.1 [95% confidence interval (CI): 0.8 1.5] for 0.18-0.30 and 1.4 (95% CI: 1.1-1.8) for more than 0.30 g/m/day. In the Netherlands Cohort Study on Diet and Cancer with 60 deaths from lung cancer, the RR was 1.06 (95% CI: 0.43-2.63) for an increase of 10 mug/m in black smoke. In the French Pollution Atmospherique et Affections Respiratoires Chroniques study cohort based on 178 deaths from lung cancer, the RR associated with an increase in exposure to 10 mug/m of total suspended particulate was 0.97 (95% CI: 0.94 1.01). A nested case-control study within the European Prospective Investigation on Cancer and Nutrition included 113 nonsmokers or exsmokers diagnosed with lung cancer and 312 controls. The RRs were 0.91 (95% CI: 0.70-1.18) for an increase in PM with diameter /=60 years). We estimated annual percentage change (APC) in melanoma incidence by regressing the logarithms of the rates and exponentiating the coefficients. Among men, overall melanoma incidence increased log-linearly in both settings, but significantly more rapidly in the west of Scotland (APC 2.8%) than Queensland (APC 1.4%). Rates of increase among Scottish men were higher for every body site and all ages than among Queensland men. Among women, overall melanoma incidence increased more rapidly among Scottish (APC 1.8%) than Queensland women (APC 0.7%). Most discrepant were trends in upper limb melanomas, which underwent large annual increases among Scottish women, but declined among younger Queensland women. Melanoma incidence continues to rise rapidly in all age groups in Scotland and among older people in Queensland. Rates of melanoma in younger people in Queensland are stabilizing, as might be expected if primary prevention campaigns were effective in reducing solar exposure. Variations in rates of change at different body sites warrant further monitoring. PMID- 18414197 TI - Genetic polymorphisms of metabolic enzymes CYP1A1, CYP2D6, GSTM1 and GSTT1 and leukemia susceptibility. AB - The genetic polymorphisms of biotransformation phase I enzymes, cytochrome P450 (CYP1A1 and CYP2D6), and phase II enzymes, glutathione S-transferase (GSTM1 and GSTT1), were analyzed in 204 healthy persons and 348 leukemia patients, who suffered from also acute lymphoblastic leukemia (ALL), acute nonlymphoblastic leukemia (ANLL) chronic myelogenous leukemia (CML), from the Han ethnic group in Changsha City of Hunan Province of China. Our results showed that the frequencies of polymorphisms of CYP1A1, CYP2D6 and GSTT1 among the groups including acute lymphoblastic leukemia, ANLL, chronic myelogenous leukemia and healthy control have no significant differences. The variation of GSTM1-null genotype alone correlated with the development of ANLL. The combined genotypes of GSTM1-null with GSTT1-null, or GSTM1-null with CYP1A1 heterozygous mutant, or GSTM1-null with CYP1A1 heterozygous mutant and CYP2D6 heterozygous mutant, or GSTM1-null with CYP1A1 heterozygous mutant, CYP2D6 heterozygous mutant and GSTT1-null were found in individuals with high risk of ANLL. All these findings suggest that GSTM1-null genotype alone or in coordination with the relevant genotypes of other metabolic enzymes might be susceptibility factors in the etiology of ANLL. PMID- 18414198 TI - Statins and cancer: a meta-analysis of case-control studies. AB - 3-HMG-CoA reductase inhibitors (statins) block the growth of malignant cells in vitro. A meta-analysis of randomized controlled trials failed to show reduced risk of cancers in statin users. Case-control studies, however, have the advantage of examining remote exposures. This study determined the association between statins and breast cancer, colon cancer, lung cancer, prostate cancer, or any cancer in case-control studies. A comprehensive search for studies published through November 2006 was performed. Twenty case-control studies (100 129 incident cancer cases) were combined to obtain a pooled odds ratio using an inverse variance method. A funnel plot did not suggest a significant absence of unpublished data. The studies were significantly heterogeneous (P<0.01), thus a random effects model was used. The pooled OR and 95% confidence intervals for statin users and cancer were as follows: any cancer 0.71 (0.56-0.89), breast cancer 0.86 (0.60-1.23), colon cancer 0.89 (0.82-0.97), lung cancer 0.75 (0.50 1.11), and prostate cancer 0.74 (0.45-1.20). In this meta-analysis of case control studies, we found a significant association between statin usage and any cancer, but when stratified by cancer type, only the association with colon cancer remained. On the basis of these results, randomized control trials with longer follow-up times than previously used are warranted. PMID- 18414199 TI - Does regional analgesia reduce the risk of cancer recurrence? A hypothesis. AB - Surgery remains the most effective treatment for most cancers, but it is usually associated with systemic release of tumour cells; additionally, preexisting scattered micrometastases commonly remain. Whether minimal residual disease succeeds in establishing itself as recurrence or metastases is primarily a function of host defence. In practice, the immune system frequently fails to neutralize remaining malignant tissues. Considerable in-vitro data and in-vivo animal studies suggest that three factors associated with cancer surgery impair cellular immunity: the stress response to tissue injury, general anaesthesia, and opioid analgesia. Regional analgesia decreases the neuroendocrine stress response to surgical tissue injury, eliminates or reduces the need for general anaesthesia, and minimizes opioid requirement. I therefore propose the hypothesis that cancer recurrence is lower after surgery with regional anaesthesia/analgesia than after surgery with general anaesthesia and opioid analgesia. Confirming this hypothesis would indicate that a minor modification to anaesthetic management reduces the risk of cancer recurrence. PMID- 18414200 TI - Does educational level determine screening participation? AB - The objective of this study was to test the hypothesis that nonparticipation in organized mammography screening is due to insufficient understanding of the information in the invitation letter by relating educational level to user pattern. Data from two Danish mammography screening programmes in Copenhagen, 1991-1999, and Funen, 1993-2001 were taken for this study. The Danish Central Population Register was used to define target groups; screened participation data were provided by the health authority, and data on highest obtained education came from Statistics Denmark. Data on all breast imaging in 2000 outside organized screening were provided by radiology clinics. Included were all women eligible for at least three screens, and participation was classified into four mutually exclusive user groups. Organized mammography screening programmes in Copenhagen and Funen, Denmark were used as field of this study. Main outcome measures were age-adjusted relative risks (RR) and 95% confidence intervals (CI) of 'never use' versus 'always use' of screening by educational level, using women with secretarial/sales education as baseline. The RR of 'never use' was 1.65 (95% CI: 1.37-1.99) in Copenhagen and 1.93 (95% CI: 1.42-2.62) in Funen for academics, 1.60 (95% CI: 1.48-1.73) in Copenhagen and 1.26 (95% CI: 1.14-1.39) Funen for women with lower primary educational level. Taking other breast imaging into account, the RR was 1.60 (95% CI: 1.32-1.95) for academics in Copenhagen, and 1.90 (95% CI: 1.75-2.07) for women with lower primary education. In conclusion, our results did not support the hypothesis that lack of understanding the information in the invitation letter explains nonparticipation. 'Never use' was not inversely associated with the level of education, but showed a U-shaped association, even when use of breast imaging outside organized screening was taken into account. PMID- 18414201 TI - Assessment of ecological regression in the study of colon, breast, ovary, non Hodgkin's lymphoma, or prostate cancer and residential UV. AB - Recent ecological studies have suggested a possible association between exposure to ultraviolet-B (UVB) radiation and reduction in the risk of various cancers; however, ecological studies are known to be subject to bias. The objective of this study was to demonstrate difficulties with the ecological approach. We conducted a multicountry ecological study using cancer incidence rates, residential UV levels, dietary intake, and different sociodemographic variables for 38 locations spanning 33 countries worldwide. The effect of residential UV exposure on cancer incidence was assessed using multiple linear regression models. The results of our multivariate analyses show no indication of an inverse association between residential UV levels and the risk of colon, non-Hodgkin's lymphoma (NHL), ovarian, prostate, or breast cancer in women. For colon cancer and NHL, a significant positive association was calculated. The rates of melanoma, which were used to examine the methods of this study, showed a strong and significant (P<0.01) association with solar radiation. Our results provide no evidence to support previous ecological results that UV exposure may reduce the risk of NHL, colon, breast, ovary, or prostate cancer. The study demonstrates the high sensitivity of ecological studies to adjustments for various confounders, and casts doubts on results of ecological analyses in this field. PMID- 18414202 TI - Polynucleotide kinase 3' phosphatase variant, dietary variables and risk of adenoma recurrence in the Polyp Prevention Trial. AB - Polymorphisms in a number of genes encoding for DNA repair enzymes have been associated with altering the function of these enzymes and increasing risk of a number of cancers, including colon cancer. We have investigated the association between a common variant in polynucleotide kinase 3' phosphatase (PNKP), a putative DNA repair enzyme, and risk of adenoma recurrence in the Polyp Prevention Trial participants. We also investigated possible interaction or effect modification between carriage of the variant allele, dietary components and risk of adenoma recurrence. Unconditional logistic regression models were used to calculate the odds ratios and 95% confidence intervals for an association between the G/T polymorphism, PNKP T5644G and risk of adenoma recurrence. We observed no association between carriage of the variant allele and risk of adenoma recurrence. Furthermore, we found no effect modification between genotype, dietary components and risk of adenoma recurrence. The PNKP T5644G variant does not seem to be involved in adenoma recurrence in the Polyp Prevention Trial. PMID- 18414206 TI - The genetics of gestational diabetes mellitus: evidence for relationship with type 2 diabetes mellitus. AB - Gestational diabetes is a major public health problem because of its prevalence, its associated complications during pregnancy, and its increased risk for type 2 diabetes later in life. Insulin resistance is one of many physiological changes occurring during pregnancy, and when insulin resistance is accompanied by pancreatic beta-cell insufficiency, gestational diabetes may develop. Several lines of evidence suggest that gestational diabetes shares a common etiology with type 2 diabetes and support the hypothesis that gestational diabetes serves as a window to reveal a predisposition to type 2 diabetes. Pregnancy is an environmental stressor that may catalyze the progression to a diabetic state in genetically predisposed women; therefore, identification of these women during pregnancy could decrease the occurrence of type 2 diabetes through targeted prevention. This review presents an overview of the genetics of gestational diabetes, focusing on human association studies with candidate genes common to both type 2 diabetes and gestational diabetes. PMID- 18414205 TI - Genetic aspects of Alzheimer disease. AB - Alzheimer disease is the most common cause of dementia and represents a major public health problem. The neuropathologic findings of amyloid-beta plaques and tau containing neurofibrillary tangles represent important molecular clues to the underlying pathogenesis. Genetic factors are well recognized, but complicated. Three rare forms of autosomal-dominant early-onset familial Alzheimer disease have been identified and are associated with mutations in amyloid precursor protein, presenilin 1, and presenilin 2 genes. The more common late-onset form of Alzheimer disease is assumed to be polygenic/multifactorial. However, thus far the only clearly identified genetic risk factor for Alzheimer disease is Apo lipoprotein E. The epsilon4 allele of Apo lipoprotein E influences age at onset of Alzheimer disease, but is neither necessary nor sufficient for the disease. The search continues for the discovery of additional genetic influences. PMID- 18414207 TI - Educating the general public about multifactorial genetic disease: applying a theory-based framework to understand current public knowledge. AB - The present article describes the application of a theory-based framework to understand current public knowledge of genetic risk factors of multifactorial diseases. The main innovative aspect is the application of E. M. Rogers' knowledge framework which distinguishes three types of knowledge: "awareness knowledge," "how-to knowledge," and "principles knowledge." We argue that distinguishing these types of knowledge allows for a more sophisticated overview of the general public. To illustrate the application of Rogers' framework, we performed a literature review of current public knowledge of genetic risk factors of multifactorial genetic diseases. Relevant articles were identified by searching the Pubmed, Web of Science, Embase, CINAHL, ERIC, and PsycINFO databases from January 1990 until January 2007 and by performing reference list and author searches. Although this review showed that current public knowledge is limited, it also showed that the knowledge framework may be a useful tool for assessing different types of public knowledge and pinpointing flaws or caveats in public knowledge with more precision and subsequently develop public health campaigns to remedy such flaws. Implications for genetic education are discussed. PMID- 18414208 TI - Long-term follow-up after diagnosis resulting from newborn screening: statement of the US Secretary of Health and Human Services' Advisory Committee on Heritable Disorders and Genetic Diseases in Newborns and Children. AB - The US Secretary of Health and Human Services' Advisory Committee on Heritable Disorders and Genetic Diseases in Newborns and Children provides guidance to reduce the morbidity and mortality associated with heritable disorders, with a special emphasis on those conditions detectable through newborn screening. Although long-term follow-up is necessary to maximize the benefit of diagnosis through newborn screening, such care is variable and inconsistent. To begin to improve long-term follow-up, the Advisory Committee has identified its key features, including the assurance and provision of quality chronic disease management, condition-specific treatment, and age-appropriate preventive care throughout the lifespan of affected individuals. There are four components central to achieving long-term follow-up: care coordination through a medical home, evidence-based treatment, continuous quality improvement, and new knowledge discovery. PMID- 18414209 TI - Array-based comparative genomic hybridization analysis of 1176 consecutive clinical genetics investigations. AB - PURPOSE: Cytogenetic investigations are useful for etiologic determinations of mental retardation, developmental delay, multiple congenital anomalies, and pregnancy complications; however, the causes remain elusive in a majority of cases despite high-resolution cytogenetic studies and multiple fluorescence in situ hybridization examinations. Array-based comparative genomic hybridization has the ability to examine the genome at a higher resolution and may yield an increased detection of genetic abnormalities. The purpose of this study was to assess the use of array-based comparative genomic hybridization in a clinical genetics setting. METHODS: DNA from 1176 patients was analyzed using a bacterial artificial chromosome array-based comparative genomic hybridization platform. All abnormal cases were confirmed by fluorescence in situ hybridization and parental studies were completed when possible. RESULTS: Of the 1176 patients included in this survey, 163 showed a genomic imbalance identified by array-based comparative genomic hybridization. Of these 163 cases, 116 had a clinically relevant genetic abnormality. A total of 9.8% (116 of 1176 cases) were determined to exhibit a causative genomic imbalance. Twenty-five of the 116 abnormal cases had a previously identified cytogenetic abnormality yielding an increased detection rate of 7.9% (91 of 1146) in cases with normal or no cytogenetics. CONCLUSION: Array-based comparative genomic hybridization increases the overall abnormality detection rate, thus improving the diagnostic potential of clinical cytogenetics investigations. PMID- 18414210 TI - Microduplications of 22q11.2 are frequently inherited and are associated with variable phenotypes. AB - PURPOSE: Genomic rearrangements of chromosome 22q11.2, including the microdeletion associated with DiGeorge/velocardiofacial syndrome, are mediated by nonallelic homologous recombination between region-specific low-copy repeats. To date, only a small number of patients with 22q11.2 microduplication have been identified. METHODS: We report the identification by array-comparative genomic hybridization of 14 individuals from eight families who harbor microduplications within the 22q11.2 region. RESULTS: We have now observed a variety of microduplications, including the typical common approximately 3-Mb microduplication, approximately 1.5-Mb nested duplication, and smaller microduplications within and distal to the DiGeorge/velocardiofacial syndrome region, consistent with nonallelic homologous recombination using distinct low copy repeats in the 22q11.2 DiGeorge/velocardiofacial syndrome region. These microduplications likely represent the predicted reciprocal rearrangements to the microdeletions characterized in the 22q11.2 region. The phenotypes seen in these individuals are generally mild and highly variable; familial transmission is frequently observed. CONCLUSIONS: These findings highlight the unbiased ability of array-comparative genomic hybridization to identify genomic imbalances and further define the molecular etiology and clinical phenotypes seen in microduplication 22q11.2 syndrome. Our findings also further support that the 22q11.2 region is highly dynamic with frequent rearrangements using alternative low-copy repeats as recombination substrates. PMID- 18414211 TI - Bacterial artificial chromosome-emulation oligonucleotide arrays for targeted clinical array-comparative genomic hybridization analyses. AB - PURPOSE: The goal of this work was to test the ability of oligonucleotide-based arrays to reproduce the results of focused bacterial artificial chromosome (BAC) based arrays used clinically in comparative genomic hybridization experiments to detect constitutional copy number changes in genomic DNA. METHODS: Custom oligonucleotide (oligo) arrays were designed using the Agilent Technologies platform to give high-resolution coverage of regions within the genome sequence coordinates of BAC/P1 artificial chromosome (PAC) clones that had already been validated for use in previous versions of clone arrays used in clinical practice. Standard array-comparative genomic hybridization experiments, including a simultaneous blind analysis of a set of clinical samples, were conducted on both array platforms to identify copy number differences between patient samples and normal reference controls. RESULTS: Initial experiments successfully demonstrated the capacity of oligo arrays to emulate BAC data without the need for dye reversal comparisons. Empirical data and computational analyses of oligo response and distribution from a pilot array were used to design an optimized array of 44,000 oligos (44K). This custom 44K oligo array consists of probes localized to the genomic positions of >1400 fluorescence in situ hybridization-verified BAC/PAC clones covering more than 140 regions implicated in genetic diseases, as well as all clinically relevant subtelomeric and pericentromeric regions. CONCLUSIONS: Our data demonstrate that oligo-based arrays offer a valid alternative for focused BAC arrays. Furthermore, they have significant advantages, including better design flexibility, avoidance of repetitive sequences, manufacturing processes amenable to good manufacturing practice standards in the future, increased robustness because of an enhanced dynamic range (signal to background), and increased resolution that allows for detection of smaller regions of change. PMID- 18414212 TI - Standards of professional practice for genetic metabolic dietitians. AB - New discoveries in the field of genetics and recent developments in newborn screening have created opportunities and challenges for genetic metabolic dietitians, placing increasing demands on dietitians to plan appropriate nutritional interventions for patients with metabolic disorders. An appropriate assessment tool must be developed to identify gaps in training and knowledge to assure that dietitians are prepared for advances in this emerging field. Using a multistage process, the Genetic Metabolic Dietitians International founders group developed a set of professional standards modeled after the American Dietetics Association's Standards of Professional Practice and the National Coalition for Health Professions Education in Genetics's core competencies. The Standards of Professional Practice for Genetic Metabolic Dietitians were validated by means of an electronic questionnaire distributed nationally to dietitians through the PRO METLAB listserv. Statistical measures were used to determine whether perceived importance was significantly associated with compliance for each of the indicators included in the Standards of Professional Practice. The Standards of Professional Practice for Genetic Metabolic Dietitians will be used to structure continuing education opportunities, guide research and evaluation, and will serve as a basis for certification and professional accreditation. PMID- 18414213 TI - ACMG recommendations for standards for interpretation and reporting of sequence variations: Revisions 2007. AB - ACMG previously developed recommendations for standards for interpretation of sequence variations. We now present the updated revised recommendations. Here, we describe six interpretative categories of sequence variations: (1) sequence variation is previously reported and is a recognized cause of the disorder; (2) sequence variation is previously unreported and is of the type which is expected to cause the disorder; (3) sequence variation is previously unreported and is of the type which may or may not be causative of the disorder; (4) sequence variation is previously unreported and is probably not causative of disease; (5) sequence variation is previously reported and is a recognized neutral variant; and (6) sequence variation is previously not known or expected to be causative of disease, but is found to be associated with a clinical presentation. We emphasize the importance of appropriate reporting of sequence variations using standardized terminology and established databases, and of clearly reporting the limitations of sequence-based testing. We discuss follow-up studies that may be used to ascertain the clinical significance of sequence variations, including the use of additional tools (such as predictive software programs) that may be useful in variant classification. As more information becomes available allowing the interpretation of a new sequence variant, it is recommended that the laboratory amend previous reports and provide updated results to the physician. The ACMG strongly recommends that the clinical and technical validation of sequence variation detection be performed in a CLIA-approved laboratory and interpreted by a board-certified clinical molecular geneticist or equivalent. PMID- 18414214 TI - Clinical genetics evaluation in identifying the etiology of autism spectrum disorders. AB - The autism spectrum disorders are a collection of conditions, which have, in common, impaired socialization and communication in association with stereotypic behaviors. The reported incidence of autism spectrum disorders has increased markedly over the past decade. In addition, a large amount of attention has been paid to these conditions among lay and professional groups. These influences have resulted in a marked increase in the number of referrals to clinical geneticists for evaluation of persons with autism spectrum disorders. The primary role of the geneticist in this process is to define etiology, if possible, and to provide counseling and contribute to case management based on the results of such investigations. In deciding upon the appropriate evaluation scheme for a particular patient, the geneticist must consider a host of different factors. Such considerations would include (1) Assuring an accurate diagnosis of autism before proceeding with any investigation. (2) Discussing testing options, diagnostic yields, and patient investment before proceeding with an evaluation. (3) Communication and coordination with the patient's medical home. (4) Assessing the continuously expanding and evolving list of available laboratory testing modalities in light of evidence-based medicine. (5) Recognizing expanded phenotypes of well-described syndromic and metabolic conditions that encompass autism spectrum disorders. (6) Defining an individualized evaluation scheme based on the unique history and clinical features of a given patient. The guidelines in this article have been developed to assist the clinician in the consideration of these factors. PMID- 18414216 TI - Not sure? Don't be afraid to ask! PMID- 18414218 TI - From novice to expert: transitioning graduate nurses. PMID- 18414219 TI - Using TIGER vision to move your agenda forward. PMID- 18414221 TI - The nurse shortage: where we stand and where we're headed. AB - Whether it's already touching your facility or its impact is still to come, this crisis will test your skills as a manager. Understanding what's driving the shortage and what's predicted will help you choose the best strategies for addressing its causes and dealing with its consequences. PMID- 18414223 TI - Policy spotlight: Health savings accounts. PMID- 18414224 TI - Using a medication protocol to improve pain management. AB - Initiative enhances prescriber options, allowing for better pain relief in patients. PMID- 18414225 TI - Point-of-care testing: issues and opportunities. PMID- 18414226 TI - Cultivating practice and education partnerships. PMID- 18414227 TI - Reduce call light frequency with hourly rounds. PMID- 18414229 TI - What's new in Shock, May 2008? PMID- 18414230 TI - Nicotinamide adenine dinucleotide phosphate (reduced form) oxidase is important for LPS-induced endothelial cell activation. AB - Activation of the endothelium plays an important role in the innate immune response. This process is associated with an increase in the production of superoxide (O2-) by nicotinamide adenine dinucleotide phosphate (reduced form; NADPH) oxidase. Our objective was to determine if O2- from NADPH oxidase contributes to activation of human umbilical vein endothelial cells by LPS as it does for TNF-alpha. We used the adhesion molecule intracellular adhesion molecule 1 and cytokine IL-8 as indicators of human umbilical vein endothelial cell activation and measured O2- production with chemiluminescence. LPS increased baseline and NADPH-stimulated O2- production. The increase was reduced by tiron, a protein kinase C inhibitor (bisindolylmaleimide I hydrochloride), the flavin inhibitor (diphenylene iodonium), and by a short interfering RNA against the p22phox component of NADPH oxidase. Inhibition of NADPH oxidase with the short interfering RNA reduced the induction by LPS of intracellular adhesion molecule 1 mRNA, protein, and IL-8 release (by enzyme-linked immunosorbent assay). The production of O2- by NADPH oxidase contributes to intracellular signaling by LPS in endothelial cells as it does for TNF-alpha and helps turn on the innate immune response in these cells. PMID- 18414231 TI - Endothelial glycocalyx damage during endotoxemia coincides with microcirculatory dysfunction and vascular oxidative stress. AB - The glycocalyx constitutes the first line of the blood tissue interface and is thus involved in many physiological processes, deregulation of which may lead to microvascular dysfunction. Because administration of LPS is accompanied by severe microvascular dysfunction, the purpose of the study was to investigate microvascular glycocalyx function during endotoxemia. Bolus infusion of LPS (10 mg kg(-1)) to male Sprague-Dawley rats elicited the development of hyporeactivity to vasoactive agents and microvascular derangements, including decreased capillary density and significant increases in intermittent and stopped flow capillaries in the small intestine muscularis layer compared with controls. LPS elicited plasma hyluronan release and reduction in endothelial surface thickness, indicative of glycocalyx degradation. Because endothelial glycocalyx is extremely sensitive to free radicals, oxidative stress was evaluated by oxidation of dihydrorhodamine in microvascular beds and levels of heart malondialdehyde and plasma carbonyl proteins, which were all increased in LPS-treated rats. Activated protein C (240 microg kg(-1) h(-1)) enhanced systemic arterial pressure response to norepinephrine in LPS-treated rats. Activated protein C (240 microg kg(-1) h( 1)) prevented capillary perfusion deficit in the septic microvasculature that were associated with reduced oxidative stress and preservation of glycocalyx. Our findings support the conclusion that LPS induces major microcirculation dysfunction accompanied by microvascular oxidative stress and glycocalyx degradation that may be limited by activated protein C treatment. PMID- 18414232 TI - Hemin arginate-induced heme oxygenase 1 expression improves liver microcirculation and mediates an anti-inflammatory cytokine response after hemorrhagic shock. AB - Microvascular failure is a major determinant for the development of hepatocellular dysfunction after hemorrhagic shock. Induction of heme oxygenase (HO) 1 may confer hepatocellular protection. Hemin arginate (HAR) induces HO-1 and protects against shock-induced organ failure. The mechanisms are not completely understood, but HO-1-mediated protective effects on the microcirculation and on the inflammatory response may contribute. Therefore, the aim of the present study was to investigate the influence of HAR pretreatment on liver microcirculation and cytokine response to assess the role of HO-1-mediated effects under these conditions. Male Sprague-Dawley rats (200-300 g; n=8 per group) were subjected to hemorrhage (MAP, 30-40 mmHg for 1 h) 24 h after pretreatment with vehicle (Ringer solution) or HAR (5 mg kg(-1)), followed by 2 h of resuscitation. The microcirculation and the redox state (nicotinamide adenine dinucleotide phosphate [reduced form; NADPH] autofluorescence) of the liver were assessed using intravital microscopy. Cytokine levels (TNF-alpha and IL-10) were quantified using an enzyme-linked immunosorbent assay. A profound induction of HO 1 was observed 24 h after pretreatment with HAR. Hemorrhage significantly reduced sinusoidal perfusion and increased NADPH autofluorescence and cytokine levels. Hemin arginate pretreatment significantly improved liver microcirculation, reduced NADPH autofluorescence, significantly increased IL-10, and tended to decrease TNF-alpha serum levels compared with shock vehicle. Blockade of the HO pathway with tin-mesoporphyrin-IX after HAR pretreatment abolished the observed beneficial effects, whereas the additional administration of the carbon monoxide donor dichloromethane reversed the tin-mesoporphyrin-IX-mediated changes. These results suggest that HAR pretreatment improves liver microcirculation and mediates an anti-inflammatory cytokine response after hemorrhagic shock through induction of HO-1 and in part through an increased carbon monoxide release. PMID- 18414233 TI - Hypertonic saline modulation of intestinal tissue stress and fluid balance. AB - Crystalloid-based resuscitation of severely injured trauma patients leads to intestinal edema. A potential mechanism of intestinal edema-induced ileus is a reduction of myosin light chain phosphorylation in intestinal smooth muscle. We sought to determine if the onset of edema initiated a measurable, early mechanotransductive signal and if hypertonic saline (HS) can modulate this early signal by changing intestinal fluid balance. An anesthetized rat model of acute interstitial intestinal edema was used. At laparotomy, the mesenteric lymphatic was cannulated to measure lymph flow and pressure, and a fluid-filled micropipette was placed in the intestinal submucosa to measure interstitial pressure. Rats were randomized into four groups (n=6 per group): sham, mesenteric venous hypertension+80 mL/kg 0.9% isotonic sodium chloride solution (ISCS 80), mesenteric venous hypertension+80 mL/kg 0.9% ISCS+4 mL/kg 7.5% saline (ISCS 80+HS), or 4 mL/kg 7.5% saline (HS alone) to receive the aforementioned intravenous fluid administered over 5 min. Measurements were made 30 min after completion of the preparation. Tissue water, lymph flow, and interstitial pressure were measured. Resultant applied volume induced stress on the smooth muscle (sigmaravi-muscularis) was calculated. Mesenteric venous hypertension and crystalloid resuscitation caused intestinal edema that was prevented by HS. Intestinal edema caused an early increase in intestinal interstitial pressure that was prevented by HS. Hypertonic saline did not augment lymphatic removal of intestinal edema. sigmaravi-muscularis was increased with onset of edema and prevented by HS, paralleling the interstitial pressure data. Intestinal edema causes an early increase in interstitial pressure that is prevented by HS. Prevention of the edema-induced increase in interstitial pressure serves to blunt the mechanotransductive signal of sigmaravi-muscularis. PMID- 18414234 TI - Human amniotic epithelial cells ameliorate behavioral dysfunction and reduce infarct size in the rat middle cerebral artery occlusion model. AB - Human amniotic epithelial cells (hAECs), having the characteristics of both embryonic and pluripotent stem cells, have the potential to differentiate into various cells. A good deal of research has explored the clinical therapeutic potential of hAECs; rat amniotic epithelial cells have been reported to ameliorate functional deficits after stroke in rats, likely due to neuronal differentiation and cytokine secretion by these cells. We isolated hAECs and transfected them with glial cell line-derived neurotrophic factor (GDNF) or enhanced green fluorescent protein (EGFP) gene using lentiviral vectors. These cells were then transplanted into the brains of rats subjected to a transient middle cerebral artery occlusion. The hAECs survived and migrated to the ischemic area of rats, and some of the transplanted hAECs expressed the neuronal marker MAP2 and the neuronal progenitor marker Nestin, together with the astrocyte marker glial fibrillary acidic protein, and hAEC-EGFP can significantly ameliorate behavioral dysfunction and reduce infarct volume of ischemic rats. By transfecting the cells with lentiviral vectors, GDNF can be stably overexpressed in hAECs, and hAEC-GDNF can more rapidly rescue the deficits of rats after middle cerebral artery occlusion compared with hAEC-EGFP-treated rats. Moreover, the nontransduced cells also had effects comparable to the GDNF-transduced cells on caspase-3 and lesion volume. Because hAECs are in unlimited supply, and their use is not encumbered by ethical arguments, hAECs have a great advantage for stem cell therapy. This model holds tremendous potential for development into wide use in cell-mediated gene therapy in the future. PMID- 18414235 TI - Exogenous cytochrome C restores myocardial cytochrome oxidase activity into the late phase of sepsis. AB - Mitochondrial dysfunction is thought to play a role in the pathogenesis of a variety of disease states, including sepsis. An acquired defect in oxidative phosphorylation potentially causes sepsis-induced organ dysfunction. Cytochrome oxidase (CcOX), the terminal oxidase of the respiratory chain, is competitively inhibited early in sepsis and progresses, becoming noncompetitive during the late phase. We have previously demonstrated that exogenous cytochrome c can overcome myocardial CcOX competitive inhibition and improve cardiac function during murine sepsis at the 24-h point. Here, we evaluate the effect of exogenous cytochrome c on CcOX activity and survival in mice at the later time points. Exogenous cytochrome c (800 microg) or saline was intravenously injected 24 h after cecal ligation and puncture (CLP) or sham operation. Steady-state mitochondrial cytochrome c levels and heme c content increased significantly 48 h post-CLP and remained elevated at 72 h in cytochrome c-injected mice compared with saline injection. Cecal ligation and puncture inhibited CcOX at 48 h in saline-injected mice. However, cytochrome c injection abrogated this inhibition and restored CcOX kinetic activity to sham values at 48 h. Survival after CLP to 96 h after cytochrome c injection approached 50% compared with only 15% after saline injection. Thus, a single injection of exogenous cytochrome c 24 h post-CLP repletes mitochondrial substrate levels for up to 72 h, restores myocardial COX activity, and significantly improves survival. PMID- 18414236 TI - Berberine inhibits cytosolic phospholipase A2 and protects against LPS-induced lung injury and lethality independent of the alpha2-adrenergic receptor in mice. AB - Acute lung injury is still a significant clinical problem having a high mortality rate despite significant advances in antimicrobial therapy and supportive care made in the past few years. Our previous study demonstrated that berberine (Ber) remarkably decreased mortality and attenuated the lung injury in mice challenged with LPS, but the mechanism behind this remains unclear. Here, we report that pretreatment with Ber significantly reduced pulmonary edema, neutrophil infiltration, and histopathological alterations; inhibited protein expression and phosphorylation of cytosolic phospholipase A2; and decreased thromboxane A2 release induced by LPS. Yohimbine, an alpha2-adrenergic receptor antagonist, did not antagonize these actions of Ber. Furthermore, pretreatment with Ber decreased TNF-alpha production and mortality in mice challenged with LPS, which were enhanced by yohimbine, and Ber combined with yohimbine also improved survival rate in mice subjected to cecal ligation and puncture. Taken together, these observations indicate that Ber attenuates LPS-induced lung injury by inhibiting TNF-alpha production and cytosolic phospholipase A2 expression and activation in an alpha2-adrenoceptor-independent manner. Berberine combined with yohimbine might provide an effective therapeutic approach to acute lung injury during sepsis. PMID- 18414237 TI - The synergistic effect of ethanol and shock insults on Caco-2 cytokine production and apoptosis. AB - Gut epithelial cells are important in orchestrating immunoinflammatory responses in the gut and may impact systemic immunocompetent cells after shock and trauma. Ethanol (EtOH) intoxication is an important etiological factor in trauma and may increase the likelihood of posttraumatic septic complications. Both EtOH and gut I/R impair intestinal barrier function. However, their combined effects on intestinal epithelial cell function and barrier integrity are unknown. Confluent CaCO2 cell monolayers were grown in a two-chamber culture system and exposed to 0.1% EtOH and/or Escherichia coli C-25 under normoxic (21% O2) or hypoxia (5% O2) followed by reoxygenation (H/R). Apical and basal compartment supernatants were collected, and TNF and IL-6 were quantitated by enzyme-linked immunosorbent assay (picograms per milliliter). CaCO2 cell integrity was indexed by apoptosis and monolayer permeability. TNF-alpha production by CaCO2 cells are greatest when incubated with EtOH and then exposed to H/R group. The apical levels of TNF production are consistently higher than basal levels, although the trend toward increased cytokine production is similar in both compartments. IL-6 production by the CaCO2 cell is also greatest when CaCO2 cells incubated with EtOH undergoes H/R. Lastly, the findings in apoptosis mirror the data of the TNF production in the apical compartment. Ethanol and H/R have a synergistic effect on cytokine production and barrier dysfunction in this model. They may also contribute to increased infectious complications and posttraumatic organ failure. PMID- 18414238 TI - Mitochondrial complex III is involved in proapoptotic BAK-induced microvascular endothelial cell hyperpermeability. AB - It has been shown that the intrinsic mitochondrial apoptotic cascade is activated in vascular hyperpermeability after conditions such as hemorrhagic shock. Studies from our laboratory demonstrated mitochondrial reactive oxygen species (ROS) formation in endothelial cells during vascular hyperpermeability. We hypothesized that the participation of mitochondrial ROS in the intrinsic apoptotic cascade results in microvascular endothelial cell hyperpermeability. The purpose of this study was to identify the site(s) of ROS formation in the mitochondrial complex(es) that leads to hyperpermeability. Rat lung microvascular endothelial cell monolayers were pretreated with inhibitors of the complex(es) (I-V) before the activation of the mitochondrial apoptotic cascade using the proapoptotic peptide BAK (BH3). Inhibitors of the xanthine oxidase, nicotinamide adenine dinucleotide phosphate (reduced form) oxidase, NOS, and cytochrome P-450 monooxygenase were also studied. The hyperpermeability was determined by the fluorescence of fluorescein isothiocyanate-albumin that leaked across endothelial cells and ROS production by 2',7& rime;-dichlorofluorescein diacetate. Cytochrome c levels were also measured. BAK (BH3)-transfected cells showed increased ROS, cytosolic cytochrome c, and hyperpermeability (P<0.05). Complex III inhibitors antimycin A (10 microM) and stigmatellin (10 microM) attenuated BAK (BH3) mediated ROS formation and hyperpermeability (P<0.05). The complex III inhibition decreased BAK (BH3)-mediated cytochrome c release. The results suggest that mitochondrial ROS formation, particularly at respiratory chain complex III, is involved in BAK-induced monolayer hyperpermeability. PMID- 18414239 TI - When Big Brother is looking out for you. PMID- 18414240 TI - An overview of the role of diet during the ageing process. AB - The UK is experiencing an ageing population but this is not matched by a similar increase in healthy life expectancy. Some of the major biological life changes associated with ageing result from the accumulation of lifelong molecular damage to cells, but many health problems are attributable to lifestyle and environmental factors. Diet is one factor believed to play a key role in the prevention and treatment of chronic diseases associated with ageing. Diets that are high in total fat, saturated fat and salt, and low in fibre, fruits and vegetables, are central to the development of CVD, type 2 diabetes and cognitive decline among elderly people. In contrast, under nutrition is also a major problem in this age group, with many elderly people having low intakes of essential vitamins and minerals. Good nutrition is essential for healthy ageing and a multi-disciplinary, life-course, approach to ageing is vital in minimising its implications for quality of life and public health. PMID- 18414241 TI - Age related macular degeneration: visual impairment with advancing age. AB - This article sets out a comprehensive outline of age related macular degeneration including the underlying disease mechanisms. Current treatment options are described together with recommendations for preventive care and support for those with visual impairment. PMID- 18414242 TI - Non-elective demand management: the renaissance of district nursing? AB - Despite the fact that the majority of district nurses are grasping opportunities to provide care to increasingly complex patients at home there have been concerns expressed recently that district nursing as a discipline is in decline. This is partly attributed to a lack of leadership and focus (Lowe, 2006). This alleged decline is set against a backdrop of rising rates of emergency admissions with associated cost pressures for many primary care organizations. This paper presents an overview of a programme which targets district nursing services at the avoidance of non-elective (emergency) admissions. The programme used a model to match staff capacity with demand to free up capacity within teams to tackle admissions from a range of client groups. Using care pathways staff were able to provide structured care for individuals at home. Teams were set targets for the number of admissions they needed to try and avoid. Within the first five months staff reduced non-elective admissions by 17% and achieved savings of 668,000 UK pounds. At a time when community services are being opened up to competition as a result of the changes proposed by "Commissioning a patient-led NHS," district nursing is capable of playing a key role in demonstrating the value of the services to commissioners. PMID- 18414243 TI - Understanding schizophrenia: a guide for newly qualified nurses. AB - Schizophrenia is a complex and severe mental illness affecting 1% of the UK's population. For those affected, interaction with a mental health educated community nurse may prove invaluable in terms of increasing the chance of an early diagnosis and preventing future relapse. NICE guidelines issued in 2002 support this theory, giving recommendations such as getting help early for those suffering and making sure that the community mental health teams work in partnership with patients and carers. The guidelines also advocate the use of atypical antipsychotics for the treatment of schizophrenia, which are available as both oral medication and long-acting injectable preparations. The use of psychosocial interventions has developed alongside medication: discussion of symptoms with a community nurse can be beneficial in identifying early signs of relapse. This information can then be fed back to the wider community mental health team. Considering that 25-40% of long-term mentally ill people refuse specialist care, combined care between mental health specialists and community nurses ensures that individuals are seen more regularly, enhancing the quality of care that they receive. Treating this condition within the community for a group of nurses with little or no experience poses professional challenges. This article is intended to inform and explain in more detail how treatment can be delivered by a broader group of individuals. PMID- 18414244 TI - Participation in community exercise classes: barriers to access. AB - Falls prevention is an important part of national health policy and while there are many causes of falls, there is increasing evidence that advocates the use of targeted balance and stability exercise training to address the risk factor of postural instability. The introduction of these exercise programmes in primary care raise questions about the most effective implementation methods that are accessible and acceptable to older people and support maximum adherence. An understanding of factors that support adherence of older people to exercise programmes is of value to community nurses to enable them to deliver health promotion advice appropriately. The purpose of this paper is to discuss the findings from interviews with older people to explore their experiences in relation to access and acceptability of local community based postural stability exercise classes. PMID- 18414245 TI - The role of the district nurse when involved in the coronial process. AB - The inquisition into the death of Diana, Princess of Wales has highlighted the role of the coroner in the investigation of the cause of death in England and Wales. The coroner provides an important safeguard to an administrative system for the registration of deaths in the United Kingdom that relies heavily on a doctor to confirm the cause of death. Coroners are required to hold an inquest where the person has died a violent or unnatural death, has died suddenly from an unknown cause or has died in custody or at the hands of the police or where an inquest is required by statute. District nurses will encounter death in such circumstances during the course of their duties and the coroner's court is the most likely court where a district nurse will be called to give evidence. It is essential you know the duties of the coroner and what is expected when giving evidence at an inquest. Following the inquiry into the case of Dr Harold Shipman the government is proposing changes to the coroner's role that will have an impact on district nurse practice (The Shipman inquiry, 2004). In a series of articles Richard Griffith and Cassam Tengnah review the role of the coroner and consider the impact of reform on district nurse practice. PMID- 18414246 TI - A rallying call for the New Year. PMID- 18414249 TI - Antimicrobial resistance or resisting responsibility? PMID- 18414250 TI - Getting to grips with migraine and headache. PMID- 18414251 TI - Minority branches in the balance. PMID- 18414252 TI - Dementia care. Part 1: guidance and the assessment process. AB - This article outlines recent guidance on dementia care and provides information on dementia, its different subtypes, the assessment process and the utility of cognitive screening tools. As dementia progresses a person may gradually lose their ability to make decisions for themselves. The Mental Capacity Act 2005 (MCA) is one of the most significant Acts to be passed in the United Kingdom, which protects people with dementia and stresses the need to advocate on behalf of this vulnerable group. The MCA is described in detail as practitioners working in the field of dementia care need to be aware of its clauses, as they are likely to require knowledge of it on a frequent basis. Dementia, delirium and depression are often mistaken for one another and useful ways to differentiate between the different conditions are given in addition to comprehensive advice about the management of people with dementia admitted to hospital with delirium. PMID- 18414253 TI - Huntington's disease. Part 1: essential background and management. AB - Huntington's disease is a neurodegenerative disorder that manifests in motor, cognitive and behavioural symptoms, gradually progressing over a number of years, if not decades. The complexity and unpredictability of the disease poses challenges for health-and social-care professionals. This article provides an essential background to the understanding and management of care for people with Huntington's disease, presenting a discussion of the main stages to highlight the symptoms and challenges associated with the disease. The article then moves on to discuss potential management and therapy issues identifying some key considerations in the treatment of the disease. Finally, the combined role of the multidisciplinary team is highlighted in providing health and social care to people with Huntington's disease as it progresses and increasing professional support is required. PMID- 18414254 TI - Outlining and defining the role of the epilepsy specialist nurse. AB - The epilepsy specialist nurse (ESN) role is complex, diverse and multifaceted, with nurses working with different patient groups in various settings. In this article, recently undertaken national audit studies into ESN services have been combined with key NHS/United Kingdom Central Council for Nursing, Midwifery and Health Visiting directives to outline the role. The main aim of the role is to provide high-quality effective assessment and care for patients/families/carers; to work collaboratively with professionals and other agencies; and to continually develop services/practices, professionals and non-professionals and themselves. Unfortunately the future of the ESN is being threatened; national audit data has identified that nurses are feeling undermined and devalued, having to take on greater workloads and more responsibilities, which they feel are not being recognized. In order to start to rectify this situation, ESNs need to collate the relevant data to prove and advertise their worth. Hopefully this vital resource will survive. PMID- 18414255 TI - Developing a venous assessment tool in IV chemotherapy administration. AB - It is recognized that the variety of intravenous (IV) chemotherapy regimens administered within the hospital setting is expanding rapidly, and this places increasing demands on nurses in relation to patient preparation and assessment. IV chemotherapy treatment is given over an extended period and requires multiple and often technically difficult peripheral cannulation attempts. To avoid the physical discomfort and potential psychological morbidity associated with repeated cannulation attempts, chemotherapy specialist nurses, in consultation with senior pharmacists, oncologists and other chemotherapy practitioners in a cancer centre, designed a venous assessment tool (VAT) that uses a scoring system to enable identification of patients who would benefit from insertion of a central vascular access device at the outset of treatment. This article describes the rationale for development of the VAT, the potential benefits of its use for both nurses and patients, and the plans for further refinement of the tool. PMID- 18414256 TI - Promoting effective continence care for older people: a literature review. AB - Recent United Kingdom guidelines have identified the need for integrated continence services within health regions. While there is evidence of improvements in community services there is little evidence that the quality of nursing care offered for older people with urinary incontinence in care settings has improved. This literature review identifies some of the underpinning issues that impact on continence promotion for older people. Despite evidence that older people suffer physical, social and psychological distress as a result of mismanaged urinary incontinence, costs of promoting continence are higher in financial terms than containing incontinence. The extent of the problem is difficult to identify in terms of how many older people are affected by different types of urinary incontinence. Nurses' attitudes are found to affect the quality of continence care delivered, and there continues to be a lack of evidence around sustainable strategies for continence promotion in care settings. PMID- 18414257 TI - The effects of biographical variables on job satisfaction among nurses. AB - A review of the literature confirms that there is no clear picture with regard to which biographical variables are strongly correlated with job satisfaction. Studies have shown small and inconsistent effects on variables, such as age, gender, education and personality, on job satisfaction. This study aimed to determine the effect of biographical variables on job satisfaction among nurses. The study used a dominant-less dominant mixed-method design and quantitative data were collected by a postal questionnaire survey. The sample was selected using stratified random sampling. The findings demonstrated no significant differences in job satisfaction for female and male nurses, and between job satisfaction and tenure. The findings did, however, confirm that nurses under the age of 35 were less satisfied than those over the age of 36; nurses with degrees were less satisfied than those without; and nurses in senior positions are more satisfied than their junior counterparts. The findings should be interpreted with caution because of the sample size and the small correlations between variables. Limitations aside, this study provides an insight into the relationship between these variables among nurses in the Republic of Ireland. It is possible that biographical variables are distal influences on job satisfaction, but further research is needed to better understand the relationship between these two sets of variables. PMID- 18414258 TI - Development of modern scientific nursing: BJN 100 years ago. PMID- 18414259 TI - The Mental Capacity Act 2005: mental capacity and mental illness. AB - In this series of articles on the Mental Capacity Act 2005 (MCA) the author now turns to the interrelation between mental capacity and mental disorder and between the Mental Health Act 1983 (MHA) (as amended by the Mental Health Act 2007 [MHA, 2007]) and the Bournewood safeguards. The article explains how the MCA and the MHA are designed to cover distinct situations: the one mental capacity; the other mental disorder and the different definitions are considered. The article also looks at the different principles which apply and the different powers available under each Act. The different forms of protection under each Act are contrasted. Because of criticism of the UK by the European Court of Human Rights in the Bournewood case, amendments have been made by the MHA 2007 to the MCA to provide protection for those incapable of making decisions who suffer from mental disorder and whose best interests require a loss of liberty. PMID- 18414260 TI - Exploring knowledge and skills on HIV in student nurses and midwives. AB - A cross-sectional survey design using a self-administered questionnaire was sent to a sample of 62 final-year student nurses and midwives to describe their knowledge of, skills related to, and attitudes towards, human immunodeficiency virus/acquired immune deficiency syndrome. Out of the 47 respondents who return the questionnaire, only 53% stated that they had received class instruction on the topic and 63.8% claimed to have increased their knowledge mainly from reading professional journals. Although only 32% said that they had cared for a patient or knew of a family member or another person with the disease, 91% indicated that they were willing to care for such patients. Overall, the respondents demonstrated positive attitudes towards this group of patients and a good level of knowledge about the subject, although some gaps were evident. However, a large majority stated that their skills to cater for the physical and psychological needs of this group of patients were deficient and would like further training. PMID- 18414261 TI - Idiopathic hirsutism: excessive bodily and facial hair in women. AB - Hirsutism is the excessive and increased bodily and facial hair growth in women in locations where hair is normally minimal or absent. It refers to the growth of hair in a pattern normally occurring only in men, and therefore primarily raises psychological, cosmetic and social concerns. Idiopathic hirsutism (IH), where the cause of excessive hair growth is unknown, is considered to be the most common form of hirsutism. It is suspected that this type of hirsutism may be familial, as there is often a family history of the condition. Women with IH will generally have normal menses and normal levels of testosterone. There are many treatment modalities that fall into two broad groups: medical and mechanical treatment. An example of a medical treatment is when an agent is used, which interferes with the synthesis of androgen at the ovarian or adrenal level, or by inhibiting the effect of androgen at the receptor level. An example of a mechanical treatment is laser hair removal, where the hair follicle is destroyed; however, much depends on the on the skill of the treating practitioner, laser type, laser spot size, skin type, hair colour, and the stage at which the hair follicles were during their hair growth cycle, and the delivered wavelength. Laser offers the fastest method of hair loss. Other mechanical treatments include electrolysis, depilatory creams, plucking and waxing. This article presents a general overview of IH, including a definition, diagnostic measures, clinical manifestations, normal and abnormal physiology, and treatment options. PMID- 18414262 TI - What's in your lunchbox? PMID- 18414263 TI - Ensuring a holistic catchphrase of reform. PMID- 18414264 TI - Shaping education for tomorrow's workforce. PMID- 18414265 TI - Recommended follow-up for ileo-anal pouch patient. AB - One of the greatest advances in colorectal surgery over the past 30-years has been the development of restorative proctocolectomy with ileal pouch-anal anastomosis for patients suffering with ulcerative colitis and selected patients with familial adenomatous polyposis. This has coincided with a proliferation of new and exciting advanced clinical roles for nurses in the United Kingdom and subsequently has led to an increase in the responsibilities and professional status of nurses. Staff development is necessary to maintain the unique contribution that nurses make to health care in the terms of practice, education and research. Nurse specialists in gastroenterology are taking their place alongside medical specialists, and more importantly establishing themselves as the principle carer in many diverse roles. However, as these nursing roles expand, a recognized framework needs to be designed, which takes into account the educational, ethical and legal issues related to accountability of running nurse led clinics, offering support, advice and follow-up for patients. This article provides nursing staff with research-based recommendations and practical guidance on running a successful nurse-led pouch clinic and follow-up service in collaboration with the consultant surgeon, gastroenterology teams and nursing staff involved specifically with the ileo-anal pouch patient. PMID- 18414266 TI - Supporting the carers of individuals affected by colorectal cancer. AB - Colorectal cancer is one of the most common cancers, and many difficulties may be experienced as a direct consequence of the disease and its treatment. These can be alleviated and/or resolved with appropriate care. Much of this care is provided informally by the individual's family and friends, whose care needs may be practical, emotional, financial, spiritual or organizational in nature. Care requirement can, as a consequence, be variable in its responsibilities, regularity and duration, since the individual diagnosed with colorectal cancer may require a wide range of support throughout their cancer journey, or have only a few temporary care needs, depending on their disease, treatment and a multitude of other personal, interpersonal and contextual factors. This highlights the importance of regular and individualized assessment. The caregiver's role is often forgotten and hence the impact it may be having upon their own health and lives gets overlooked. Yet, a carer's ability to manage this role has an important bearing not only on patient experience but also on the level of professional involvement. To perform this role satisfactorily carers need to be prepared, supported and also assessed at key points in the patient's cancer journey. Nurses have a professional obligation to help carers by first preparing them, then maintaining sufficient contact to be able to proactively provide practical support and additional health care as needed. This article focuses on these carers and highlights the scope, associated challenges and possible consequences of the caring role. It also seeks to stimulate reflection of how current service provision for carers might be improved. PMID- 18414267 TI - Causes and management of indwelling urinary catheter-related pain. AB - Urinary catheters can be introduced into the urinary bladder either per urethra or via the suprapubic route; this article examines indications and contraindications to these routes. Catheter pain is subdivided into pain experienced as the catheter is passed, while in situ, and on removal. Relating to pain felt on insertion, risks associated with local anaesthetic/antiseptic gel and the occurrence of paraphimosis are discussed. Once in situ, the type of material used to manufacture the catheter, pressure on the urethra caused by a large Charriere size catheter, or from drainage bag traction, leading to discomfort and possible tissue damage are examined and solutions suggested. Catheter-associated meatal trauma in men and urethral post-menopausal discomfort in women are addressed; likewise, catheter cramp due to bladder spasm or catheter blockage, and interventions are recommended. Encrustation and 'cuffing' may cause pain during catheter removal and again, advice is given. Lastly, the possibility of pain due to fear/non-acceptance of the catheter is raised. In conclusion, once identified, the cause of catheter pain is often treatable, but should not be regarded negatively or ignored, as it warns of potentially harmful conditions and allows intervention before permanent damage can occur. PMID- 18414268 TI - Dementia care. Part 2: understanding and managing behavioural challenges. AB - Management of dementia should focus on the maintenance of function and independence for the person with the disease. Non-pharmacological interventions to manage dementia include reality orientation, aromatherapy and validation therapy. Non-pharmacological ways of managing challenging behaviour are recommended in current guidance. Drug treatments, which may stabilize the symptoms of Alzheimer's disease or slow down further progression for a time, have been available in the United Kingdom since 1997 and may help some people with dementia. There are high levels of stress, distress and psychological illness in family caregivers of individuals with dementia, and practitioners are well placed to signpost carers to appropriate support organizations. PMID- 18414269 TI - The Mental Capacity Act 2005: children and young persons. AB - In this final article of the series the author considers the extent to which the Mental Capacity Act relates to children. While the Act applies to those over 16 years, certain provisions only apply to those over 18 years: these are advance decisions, and the power to set up or be appointed under a lasting power of attorney. However, there are provisions which could apply to a child under 16 years old, if he or she is unlikely to have the requisite mental capacity at 16 years. There are powers set up to enable easy transfer of proceedings between the Court of Protection and the Family Division of the High Court and vice versa. PMID- 18414271 TI - Nursing care of tuberculosis patients: BJN 100 years ago. PMID- 18414270 TI - Nursing care and management of patients with sarcoidosis. AB - Sarcoidosis is a rare genetic autoimmune disease that can attack any organ and tissue type in the body. Although it has been recognized for over 100 years the aetiology remains unknown. The group most at risk is black females under the age of 40, but no group is immune to it. The disease is commonly asymptomatic at diagnosis and often resolves without medication; however, where it does manifest it can become chronic and can be fatal. Corticosteroids are the preferred treatment although there are alternative approaches. The disease can develop in different systems simultaneously, and sarcoidosis of the lungs, skin, eyes, heart, the central nervous and musculoskeletal systems are explored in this article. The practicalities of nursing are discussed, and the work of the nurse Imogene King (1981) is shown as a useful strategy to address chronic sarcoidosis. PMID- 18414272 TI - Huntington's disease. Part 2: treatment and management issues in juvenile HD. AB - Juvenile Huntington's disease (JHD) is a rare condition, with only about 5-10% of Huntington's disease cases occurring in individuals under the age of 20 years. Symptoms of JHD include, for example, rigidity, stiffness, awkwardness in walking, and speech difficulty. JHD is caused by an inherited gene mutation that is localized to the short arm of chromosome 4. There is no cure for the condition, and it is currently managed through symptomatic treatment and supportive care. Being an inherited condition, those involved in the care of a child with JHD need to be aware of the impact that the disorder will have on the whole family. PMID- 18414273 TI - Nurses' knowledge of mouth care practices. AB - AIM: to assess nurses' knowledge and practices concerning oral care on surgical and medical wards in a district general hospital in Ireland. METHODS: a survey of all qualified nurses on three medical and three surgical wards was carried out. Two hundred questionnaires were distributed and 50 were returned with 43 of these rendered useable. RESULTS: the majority of nurses (90%) viewed oral care as an important aspect of nursing care. However, nurses lacked adequate knowledge of oral care practices, and experienced problems attending to oral care as a result of time constraints, lack of oral care packs, patients being confused or uncooperative, lack of toothpaste and toothbrush, limited education and low status attached to oral care. Although most nurses (over 70%) had received education on oral care, this was of short duration. Sixty-four per cent did not use an assessment tool to measure the condition of the patient's oral cavity, while 73% believed that an assessment tool would be important to measure a baseline of the condition of the patient's oral cavity. CONCLUSION: although oral care is an essential component of quality nursing care, it appears to be given low priority by some nurses. This study indicates the need for educational updates for qualified nurses, adequate supply of oral care equipment and promotion of formal assessment tools usage in the hospital setting. PMID- 18414274 TI - Managing the job lottery. PMID- 18414276 TI - The lion, the witch and the petri dish. PMID- 18414277 TI - Learning disabilities is bigger than a cursory review. PMID- 18414278 TI - Stress and burnout in forensic mental health nursing: a literature review. AB - Forensic mental health nurses who work with patients who have severe and enduring mental health needs have been identified as at risk of suffering from occupational stress, and even developing burnout syndrome. Therefore, this article reviews the available literature on stress and burnout in inpatient forensic mental health nursing to identify the stressors and to highlight recommendations. From the review, the main stressors placed on forensic nurses are identified as interprofessional conflicts, workload, and lack of involvement in decision-making. Recommendations to reduce stress and burnout for nurses within this specialty are highlighted. These are identified as follows: staff should have easy access to support systems including clinical supervision; managers should foster an open and honest culture to enable staff members to express their feelings openly or in confidence and learn how to deal with their frustrations; and staff should be encouraged to rotate wards to increase personal and professional development and reduce boredom and apathy. Furthermore, staff should be provided with, and encouraged to undertake, continuing professional development which may include psychosocial interventions training. PMID- 18414279 TI - The die has been cast? Rediscovering the essence of psychiatric nursing. AB - Examination of the international literature that focuses on the historical 'developments' in psychiatric nursing shows a 'back and forth' shift in emphasis from interpersonal/therapy-based models to a concentration on pathophysiology, pharmacology and biological causation. Within this context, a number of ontological questions appear with conspicuous regularity and these are worthwhile considerations as they are tied to the very nature of psychiatric nursing. Examination of the relevant literature reflects this historical duality and indicates the existence of (at least) two versions of psychiatric nurse, each with a distinct emphasis. Given this documented duality, it is the duty of each nurse to ask themselves: What type of psychiatric nurse am I? What type do I want to be and what type would I want participating in the care of someone I love? Consequently, this article draws on bodies of literature pertaining to the extent of mental health problems, the views of mental health service users, and the different types of psychiatric nurse, in order to assist nurses in considering these questions. In the light of the clear emphasis evident within contemporary mental health policy and psychiatric nursing curricula, the article concludes that it may not be possible to reconcile the activities associated with both types of psychiatric nurse. PMID- 18414280 TI - Providing effective support for patients facing disfiguring surgery. AB - Surgery for head and neck cancer can leave patients with obvious facial disfigurements. Some individuals adjust remarkably well to their altered appearance and require very little emotional support. Conversely, individuals with poor coping skills or limited support from those close to them can become socially isolated and depressed. Nurses, caring for patients during their hospital stay, are in the ideal position to assess how they are emotionally adapting to having a disfigurement. By listening to the patient, and their family's concerns, nurses can make a real difference to their recovery and quality of life. Through facilitating the development of the individual's practical coping skills and confidence in social situations, nurses can help patients to start to constructively adjust to their new appearance. It is crucial to put patients and their families in touch with ongoing support before they go home. PMID- 18414281 TI - The personal tutor's role in pre-registration nursing education. AB - Providing support for students has always been an inherent part of the lecturer's role. Widening access to students from backgrounds not traditionally associated with higher education has necessitated a review of the personal tutor system, as potential students may not be equipped with the academic skills to meet the demands of higher education. Ten randomly selected lecturers in one institute of higher education were interviewed to elicit their views and experiences of the personal tutor's role. The findings showed that the role of the personal tutor was considered both demanding, as students have increasingly different needs, but also fulfilling. The provision of pastoral care was viewed as integral to the role of the personal tutor, although most lecturers reported that they adopted an all-encompassing model of support that ranged from providing academic, professional, psychological and financial advice. Time factors and workload were identified as important structural constraints. PMID- 18414282 TI - Severe allergic reaction: management of anaphylaxis in hospital. AB - Anaphylaxis is an acute, severe, hypersensitivity reaction that can lead to asphyxia, cardiovascular collapse and cardiac arrest. This reaction is sudden, severe, and involves the whole body. Common causes include foods such as nuts, shellfish, dairy products and eggs. Non-food causes include bee/wasp stings, latex and drugs, e.g. penicillin. Common clinical features include urticaria, angioedema, respiratory distress and shock. Summoning expert help, reclining the patient flat, administering high concentration oxygen, and administering intramuscular adrenaline are key aspects of the nursing management of anaphylaxis in hospital. The aim of this article is to understand the management of anaphylaxis in hospital, with particular reference to national consensus guidelines. PMID- 18414283 TI - The Mental Capacity Act 2005 and decision-making: code of practice. AB - Continuing the series of articles on the Mental Capacity Act 2005, which for the most part came into force in October 2007, this article considers the significance of the Mental Capacity Act 2005 Code of Practice (Department of Constitutional Affairs, 2007), and its implications for nursing staff. The Secretary of State is required to provide a code or codes of practice on the Act covering specified areas. A draft code was prepared and was revised that came into force on 1 April 2007. The article sets out who is bound to follow the Code of Practice and the consequences if it is ignored. Informal (unpaid) carers are not required by the Act to follow the Code but it is hoped that in practice they will do so. The Ministry of Justice (replacing the Department of Constitutional Affairs) is responsible for monitoring the Code and will revise it from time to time. PMID- 18414284 TI - A pretty little game of make-believe. PMID- 18414286 TI - Achieving dignified care. PMID- 18414285 TI - Medication management in older adults: a critique of concordance. AB - This article shows that the terms compliance, adherence and concordance are used interchangeably in the medication management literature. As such, it is argued that nurses should focus on those interventions that are demonstrably effective in enhancing medication management for the older adult rather than attempt to make sense of a meaningless ideal. In this article the concepts of concordance, compliance and adherence are first critiqued and it is then argued that all the terms remain valid for practical purposes. That is, a literature search of all the terms is required to comprehensively discuss medication management. Focus then switches to factors that have been shown to be beneficial as well as detrimental to medication management in older adults. While many factors appear to correlate with good and bad management of medication the conclusion is that individual, tailored approaches are most effective. For the purpose of this article, the term 'older adult' refers to those over 65 years where not otherwise specified. PMID- 18414287 TI - Everyone likes a quitter. PMID- 18414288 TI - Promoting presumed consent in organ donation. PMID- 18414289 TI - Paradigms of nursing: which will it be? PMID- 18414290 TI - Psoriasis: characteristics, psychosocial effects and treatment options. AB - Psoriasis is a complex chronic non-infectious inflammatory skin disease with many different presentations. The classic presentation is of well-defined red plaques with silver scale. The characteristic scale makes the disorder highly visible and intrusive on the patient's lifestyle. The visible nature of the disease ensures that psoriasis has both physical and psychosocial effects. In normal skin, epidermal cell reproduction and proliferation takes 28 days. In psoriasis, this process is considerably accelerated to approximately four days, resulting in the deposit of immature cells on the skin. While the exact cause of this process is unknown, certain environmental and genetic factors are known to be provoking factors. Disease management will be dependent on disease severity, psychosocial effects and the patient's lifestyle. To effectively treat this disease the nurse must be skilled in psoriasis management, and in patient education and motivation. This article reviews the characteristics, aetiology, psychosocial effects and treatment strategies of psoriasis. PMID- 18414291 TI - Epidermolysis bullosa. Part 1: causes, presentation and complications. AB - This article is the first in a series of three focusing on the causes, clinical presentation, complications and care of adult patients affected by epidermolysis bullosa (EB), a group of rare genetic skin fragility disorders. Although the condition is rare, in some cases it presents extreme challenges both to those affected and those involved in the care of the EB patient; therefore, these articles may have relevance for other long-term disorders. While there is a wealth of information regarding the 'science' of EB there is dearth of information regarding the care of the adult EB patient, and this series of articles will endeavour to fill that gap. This article focuses mainly on those patients affected with the most severe form of EB found in the adult group, recessive dystrophic epidermolysis bullosa; with the part two looking at the care of the adult with EB from the nursing perspective, including wound management, and the experiences of a specialist EB psychotherapist being presented in the final article of the series. Readers will thus have an opportunity to gain an overall view of this difficult condition. PMID- 18414292 TI - Current clinical overview of cutaneous melanoma. AB - This article reviews current evidence on epidemiology, diagnosis and management of cutaneous melanoma. Incidence of cutaneous melanoma is rising in all Caucasian populations across the world; thus, melanoma represents a significant public health burden. Although, incidence of melanoma is in continuous increase, a decrease of mortality and improved survival has been observed in most western European populations. Clinical characteristics of four major types of melanoma (superficial spreading, nodular, lentigo maligna melanoma and acral lentiginous melanoma) have been described. Surgical removal of melanoma remains the standard care in all primary melanomas. Current evidence suggests use of 1 to 2 cm excision margins. Wider margins may be necessary in patients with thicker melanomas with higher risk for local recurrence. In the treatment of regional lymph nodes elective lymphadenectomy has been surpassed by the sentinel lymph node biopsy (SLNB). However, although prognostic value of SLNB has been confirmed, its therapeutical benefit still needs to be evaluated. Currently there is no standard adjuvant therapy for melanoma although interferon-alpha has been the most widely used treatment in the adjuvant setting. The role of metastasectomy (removal of distant metastases) is still controversial. Chemotherapeutic agents have a limited activity in patients with metastatic melanoma with response rates up to 25%. Although different vaccines have been tested in melanoma patients their role still remain to be established in phase III trials. Progresses in molecular biology and genetics of melanoma may lead to the development of novel melanoma therapies. PMID- 18414293 TI - Dementia care. Part 3: end-of-life care for people with advanced dementia. AB - End-of-life care issues for people with advanced dementia have only recently been addressed in guidance. There appear to be barriers to accessing good palliative care for people in the terminal phase of the disease. The reasons for this are multifactorial, but may be attributed to factors such as dementia not being recognized as a terminal disease like cancer, problems in recognizing the symptoms of terminal dementia, and decision-making conflicts between family caregivers and other health and social care providers. This article highlights common symptoms of advanced dementia, and the need for a palliative care approach. It also addresses specific issues in both caring for people with dementia at the end of their lives and in supporting carers. PMID- 18414294 TI - Sporicidal activity of two disinfectants against Clostridium difficile spores. AB - The sporicidal activity of an odour-free peracetic acid-based disinfectant (Wofasteril) and a widely-used dichloroisocyanurate preparation (Chlor-clean) was assessed against spores of the hyper-virulent strain of Clostridium difficile (ribotype 027), in the presence and absence of organic matter. In environmentally clean conditions, dichloroisocyanurate achieved a >3 log10 reduction in 3 minutes, but a minimum contact time of 9 minutes was required to reduce the viable spore load to below detection levels. Peracetic acid achieved a >3 log10 reduction in 30 minutes and was overall significantly less effective (P<0.05). However, in the presence of organic matter - which reflects the true clinical environment - there was no significant difference between the sporicidal activity of dichloroisocyanurate and peracetic acid over a 60-minute period (P=0.188). Given the greater occupational health hazards generally associated with chlorine releasing agents, odour-free peracetic acid-based disinfectants may offer a suitable alternative for environmental disinfection. PMID- 18414295 TI - Skills for caring: valuing knowledge of applied science in nursing. AB - The purpose of this article is to consider the implications of over-emphasizing the role of the nurse in terms of interpersonal relationships and emotional engagement. It seeks to explain why nurses are frequently more comfortable with the relational aspects of their work and less comfortable with their many tangible contributions. It examines why nurses often undervalue their considerable knowledge of applied science. It also evaluates why a change in attitude is necessary, and why nurses should enhance and give more prominence to their technical knowledge and skills. Finally, it includes a brief examination of current trends in nurse recruitment and retention, and their implications for the future of the profession in respect of the balance between interpersonal and technical skills. PMID- 18414296 TI - Law in focus: incorporating legislation and guidelines into practice. AB - In this new series on specialist healthcare law, the aim is to explore and review the ways in which legislation and government white papers have begun to affect clinical practice. This article discusses how media in the United Kingdom have taken to themselves the task of reporting widely on all aspects of healthcare and its delivery. The effect of this public portrayal is compounded by the disparity between what nurses believe their profession is, and the way in which our political masters have portrayed the profession. PMID- 18414297 TI - Huntington's disease. Part 3: family aspects of HD. AB - Research into the experience of the Huntington's disease (HD) family caregiver has established that HD carers experience a number of unique obstacles within their caregiving role. This appears to be due to the chronic nature of the disease, both in terms of genetic inheritance and the prolonged disease process itself. Moreover, due to the complex, physical, neurological, psychiatric and genetic elements of HD, service provision may often be unsuitable leaving family members burdened with the main responsibility of care. The complex nature of HD requires service providers, researchers and policy makers to consider each case on an individual basis, thus tailoring service provision to the user and their family's unique requirements. PMID- 18414298 TI - The art of medicine giving: BJN 100 years ago. PMID- 18414300 TI - Keeping fit for the game. PMID- 18414299 TI - Guided reflection: a discussion of the essential components. AB - Guided reflection offers practitioners the confidence to analyse and confront their practice with a critical companion in a safe environment. It is well documented that this process has the potential to liberate practitioners from oppressive constraints of practice. When guided reflection works well the consequences are manifold; however, in the author's experience the practice has raised some critical questions for novice guides willing to supervise students' clinically based reflections. Questions from guides have emerged, such as: Where do we start? How do reflectors choose their guide? What happens if things go wrong? In this article, four practical steps for guides beginning guided reflection are suggested. PMID- 18414301 TI - Coding the path for practice accountability. PMID- 18414302 TI - A life unlike any other; human rights for some? PMID- 18414303 TI - Diabetes and chronic kidney disease: a complex combination. AB - Diabetes and chronic kidney disease (CKD) are two disease processes that remain with patients from diagnosis to the end of their lives. Both are destructive conditions that must be diagnosed early to prevent longer-term complications, such as retinopathy and neuropathy. Diabetes remains the single most important cause of kidney failure. Diabetes in hospital accounts for 10% of all admissions, but this figure is much higher within renal medicine with diabetes significantly increasing length of stay, with excess bed days suggested at 20%. Inpatient care is currently the largest single component of medical costs for diabetes, while diabetes costs 10% of the NHS budget. The number of patients with both diabetes and CKD are increasing, and to help slow down the progression to complications patients must receive education on how to control their diabetes. Training packages are designed to facilitate this educational process but the NHS needs to invest in refresher courses as the educational structure needs to support lifetime learning. It is vital from first referral that all patients lead in the decisions made about their health. Healthcare professionals have a duty of care to ensure patients are given clear, concise and accurate information in language they understand with the use of medical terminology and jargon limited. Patients must then be allowed to gather their thoughts and ask any further questions before supporting them in the choices that they make. PMID- 18414304 TI - Dialysis access and the impact on body image: role of the nephrology nurse. AB - End-stage renal disease is a chronic condition, without cure, requiring dialysis therapy to maintain life or transplantation for those fortunate enough to receive a kidney. To commence dialysis (peritoneal dialysis or haemodialysis), access is required in the form of a fistula, vascular catheter or peritoneal catheter, and changes in body image will ensue, no matter what treatment option is selected. Renal transplantation, the treatment of choice for many patients, is also associated with body image issues. Despite these problems, the role of the nurse in managing body image problems in the renal population is rarely discussed. The aim of this article is to outline the concepts of dialysis and body image, and discuss the role of the nephrology nurse at the authors' current place of work. It highlights suggestions on how renal nurses can prepare and educate patients regarding changes in body image following a diagnosis of end-stage renal disease, and emphasizes the need for future research in this area. PMID- 18414305 TI - The development of 'village nurses' reports: BJN over 100 years ago. PMID- 18414306 TI - Mild cognitive impairment: a review and nursing implications. AB - The concept and classification of 'mild cognitive impairment' is disputed territory. This article reviews the literature on mild cognitive impairment and suggests that despite the complexity, early identification of dementia presents an ideal opportunity for nurses to work together with other statutory and voluntary partners, to provide support and intervention for people living with this stage of the condition and their families. Practice that is evidenced through individual and family history, coupled with a range of psychosocial interventions, including a quality-of-life appraisal, would seem an ideal way to structure support at this transitional point in time. A fictitious case study is included which demonstrates that stress and negative adjustment are just as likely to occur early in the course of dementia as later on in the condition. PMID- 18414307 TI - Preceptorship: facilitating student nurse education in the Republic of Ireland. AB - Nursing is a practice-based discipline and clinical practice is considered to be an integral part of nurse education. The pivotal role that preceptors play in the socialization, teaching and assessing of student nurses, is vital in helping students to integrate theory with practice. However, some evidence suggests that there can be minimal contact between the stakeholders involved in the students' education process, namely the university lecturers and preceptors. The focus of this article is on exploring the literature on models of support for student nurses with particular emphasis on the concept of preceptorship. PMID- 18414308 TI - Patients' experiences of privacy and dignity. Part 1: a literature review. AB - This article reviews the literature on patients' privacy and dignity and concludes that interest in the topic goes back to psychiatric patients in the 1960s, with the 1990s seeing a shift to all hospitals. A number of databases were used to obtain information for the literature review. Findings revealed United Kingdom (UK) government policies designed to raise the profile of patients' privacy and dignity. UK research on patients' privacy/dignity is limited, although there is universal interest. While a variety of methodologies have been used in previous research, and there is consistency in findings, more research is indicated. Previous research spanned topics such as elderly patients, medical and surgical patients, palliative care, rehabilitative settings and child-bearing women. A conceptual framework and definitions of privacy and dignity were examined. Although healthcare professionals and patients attach importance to patients' privacy, there is insufficient understanding of the problem. Limitations of previous research have been discussed. PMID- 18414309 TI - The duty of care and political expectations. Part 1: a literature review. AB - A consequence of the Government's agenda for nursing is that registered nurses are increasingly required to regard themselves as autonomous practitioners. It is therefore necessary to re-visit the legal principles involved so as to be able to identify where safe practice can be incorporated into the new innovative ways of nursing. The purpose of this article is to identify and reflect upon the ways in which nurses are incorporating these principles into practice, and secondly, to reflect on the question of accountability as expressed through the law and through the professional regulatory body's code of professional conduct. PMID- 18414310 TI - Nursing management of patients with a chest drain. AB - Chest drains incorporate the use of a one-way valve to drain fluid or air from the pleural cavity. The valve prevents back-flow of air and fluid into the pleural cavity. They are indicated for use when collections of fluid or air are present in the pleural space, and by draining the collection they restore efficient gaseous exchange. Little has been written on the nursing management of chest drains and the literature highlights a lack of national standardized guidelines for due to the range of thoracic conditions encountered by clinical staff. Themes such as pain management and mechanism of breathing occur frequently; however, there is a lack of up-to-date literature for the nurse to refer to. This article examines the nursing role in chest drain management from insertion to removal and includes aspects of pain management and features of a functioning chest drain. PMID- 18414311 TI - The healing power of prayer and its implications for nursing. AB - Prayer is widely acknowledged in both ancient and modern times as an intervention for alleviating illnesses and promoting good health. There is increasing attention on prayer in health care, in both popular and serious discourse. Advocates exalt the healing power of prayer in health care, while critics are sceptical about this claim and its healing potential is put down to coincidences or its placebo effect. Consequently, a variety of empirical studies have attempted to test its effect scientifically with no conclusive results. There is evidence to suggest that some patients and healthcare practitioners believe in the healing power of prayer. Nurses may be called upon to pray with or for patients as part of holistic care. This article sets out to explore the role of prayer in healing and its implications for nursing. To achieve this aim, this article provides a review of discourses and evidence on the power of prayer in healing. Its implications for nursing are highlighted with some suggestions on how to respond to patients' spiritual needs. It is concluded that, although the evidence on the healing power of spirituality is inconclusive, there are indications that it has potential for the health and wellbeing of both patients and nurses. PMID- 18414312 TI - Shouldn't the dangers of insulin be controlled better? PMID- 18414319 TI - Should we tag people with dementia? PMID- 18414320 TI - Symptom pathways from referral to death: measuring palliative care outcomes. AB - This article reports on the first stage of a process to develop a clinical performance indicator for a community-based palliative care service that may inform the development of an agreed set of indicators for the whole sector. The study explores whether symptom data that are routinely collected to plan and monitor the care being provided by a community-based palliative care service also have the potential to be used to audit care against standards, compare services, and inform funders or purchasers. The article looks at the symptom distress scores of patients over the whole episode of their care from referral to death, by constructing pathways for each symptom (pain, insomnia, appetite, nausea, bowel, breathing problems, fatigue and pain). How these data may provide a multi purpose measure of clinical outcomes is then discussed. It is concluded that, following further research to establish the symptom assessment scale's reliability and validity in different settings, it should be considered for universal adoption by palliative care services. PMID- 18414321 TI - The lived experience of patients with pleural mesothelioma. AB - This paper reports on a research study of five patients diagnosed with mesothelioma. The study used a phenomenological approach to explore patients' lived experience using in-depth interviews. The findings identify that patients have many unmet psychosocial and emotional needs and that there was a lack of information provided to patients about specialist supportive and palliative care services. A number of the patients found specialist supportive care by chance rather than by referral. In addition, patients were involved in complex medico legal matters in relation to asbestos exposure, and this was an additional burden for them and their spouse or carer. A feeling of social isolation was also reported and a number of patients would welcome the opportunity to meet with other people in the same situation as themselves. In conclusion, there is a lack of attention to the emotional needs of this group of patients, which means that supportive care resources are not being accessed in a timely and flexible manner. PMID- 18414322 TI - Electronic surveillance and tagging people with dementia. PMID- 18414323 TI - Holistic assessment of cancer patients' pain: reflections on current practice. AB - This article explores the holistic assessment and management of pain for palliative care patients through the use of a case study. Using literature searching and reflection on current local practice, issues within pain assessment and management for cancer patients are identified. These issues include communication, opioid misconceptions, disease progression associated with increased pain, and the need to address all components of cancer patients' total pain. Reflection on practice has highlighted that education, effective communication and multidisciplinary collaboration, and adaptation of an existing assessment tool to develop a multidimensional holistic approach to assessing pain, have the potential to improve quality of life for palliative care patients and their families. PMID- 18414324 TI - Continuing bonds: a human response within paediatric palliative care. AB - The concept of continuing bonds (CB) is a human response to health and illness within paediatric palliative care. This literature review aims to: define and explain the significance of CB; describe the history of the concept; critically analyze the literature related to CB; and make recommendations for future research. Exploration of CB as a response to life-threatening illness and to losing a child is significant in terms of improving care for children and their families living with life-threatening illness. This paper focuses on children with a life-threatening illness, but CB also needs to be studied with other populations, such as those with life-limiting conditions. PMID- 18414325 TI - Quality of life in palliative care: being at ease in the here and now. AB - Quality of Life (QoL) is an important issue for users of hospice services and may be assessed in a quantitative way. This study set out to evaluate the effectiveness of the Schedule for the Evaluation of Individual Quality of Life- Direct Weighting (SEIQoL-DW). A sequential mixed methods design was used to examine both quantitative and qualitative factors relating to quality of life. Five participants were recruited from an inpatient specialist palliative care (hospice) unit in East Anglia. Each participant completed the SEIQoL-DW followed by unstructured interviews, which were analysed using Giorgi's (1985) psychological phenomenological method. It was found that SEIQoL-DW provided an opening or 'window' onto participants' QoL as they travelled from their past to their future. Themes emerging from the unstructured interviews suggest that QoL was about 'being at ease' in the 'here and now' and was influenced by past experiences and future expectations. Existential and spiritual issues, which were not all captured by the SEIQoL-DW, were of more importance than physical symptoms. This needs to be taken into account when caring for those nearing the end of their lives and particularly when using medications that have an adverse effect on mental acuity. PMID- 18414326 TI - Quality of life in patients with cancer. PMID- 18414327 TI - The lived experience of closeness in partners of cancer patients in the home care setting. AB - At present there are few data available on the experiences of closeness in partners of patients with advanced cancer. The aim of this study was to explore the lived experience of closeness in partners of patients with advanced cancer in palliative home care. Ten partners of patients admitted to a palliative, hospital based home care unit participated in this study. Tape-recorded interviews were conducted and Giorgi's phenomenological analysis was used. Three themes emerged: dimensions of closeness, such as togetherness, sharing and daily concern; prerequisites for closeness; and obstacles that hindered closeness. In essence, closeness was described as a changing process that was expected to end. This study has revealed the importance of closeness in the partners of patients with advanced cancer. Experiencing closeness is of great importance when the disease is no longer curable and time is limited. Partners need professional help to facilitate this experience. PMID- 18414328 TI - Diagnosis and management of neuropathic pain in palliative care. AB - Neuropathic pain can be distressing and difficult to treat, and remains a problem for a significant proportion of palliative care patients. This article considers the identification and assessment of neuropathic pain, and highlights some of the challenges specific to the palliative care population. Further discussion includes definitions, pathophysiology and implications for nursing practice. PMID- 18414329 TI - Is a STAS-based tool valid to triage patients at a specialist palliative care inpatient unit? AB - Many tools exist to assess the symptoms and needs of palliative care patients, but no tool has been validated to prioritise patients referred for specialist inpatient palliative care. The aim of this study was to produce and validate such a tool. A prospective pilot study produced a Support Team Assessment Schedule- (STAS-) based tool--the Admission Assessment Tool (AAT)--and compared this with the existing system of triage at the Marie Curie Hospice, Edinburgh. Validity of the tool was not confirmed and the tool was modified and re-evaluated. One hundred and twenty-seven consecutive patients referred to the hospice received three AAT scores: from the bed manager; the admitting doctor; and the admitting nurse. The hospice's multidisciplinary team assessed the urgency of each patient's admission. The overall correct classification rate was approximately two thirds, but false positive rates were high and there was poor inter-rate correlation. It is concluded the AAT has not been validated. PMID- 18414330 TI - Sleep disturbance in patients with advanced cancer. AB - Sleep is a complex process, acknowledged to be essential for health and wellbeing. Sleep disturbance is reported to be a significant problem for patients across the cancer care trajectory, including those in the palliative phase of illness. This paper reviews the literature specific to sleep disturbance in patients with advanced cancer. The Human Response to Illness model, which is consistent with the central tenets of palliative care philosophy, provides a valuable framework to examine and organize the current knowledge related to sleep disturbance. The four perspectives of this biopsychosocial nursing model- physiology, pathophysiology, behavioural and experiential, as well as the personal and environmental factors--offer a broad perspective to better understand this multidimensional symptom and create a strong foundation for nursing care and future research directions. PMID- 18414331 TI - Palliative care in nursing homes: a comparison of high- and low-level providers. AB - The purpose of this study was to explore staff perceptions and concerns about the use of palliative care services in the nursing home setting. Six administrators from nursing homes were purposively selected for key informant interviews. Four common themes emerged, including issues related to the culture of care, the model of care, the relationships with hospice partners, and the role of staff. Recognition of staff perceptions is an important first step in improving the utilization of palliative care services. Staff insight provided clarification related to impediments in promoting a culture of care that was person-centered and relationship-based. We conclude by identifying the solutions for raising the level of dialogue to promote palliative care practice in the nursing home environment. PMID- 18414332 TI - A question of evidence: decision-making in palliative care nursing. AB - Nurses are urged to integrate research evidence into their practice. Current perspectives of evidence-based practice promote a hierarchy of empirical evidence. Notwithstanding the contribution of empirical knowledge, this approach constrains our understanding of the value and contribution of other forms of knowledge. Shifting to a model of 'evidence informed practice', where multiple forms of knowledge are considered and valued, may better reflect the complexities of end-of-life care and offer a fuller understanding of palliative care nursing best practice. PMID- 18414333 TI - Innovation: a core requisite for palliative nursing. PMID- 18414334 TI - Diagnosis and management of malignant spinal cord compression: part 1. AB - Malignant spinal cord compression (MSCC) is a particularly challenging area of cancer care where early diagnosis and expert multidisciplinary care and rehabilitation are paramount in optimising quality of life for the affected individual. The effects of MSCC can range from minor sensory, motor and autonomic changes to severe pain and complete paralysis that significantly affects the remainder of a patient's quality of life. When caught early, the symptoms of MSCC can be prevented, minimised or possibly reversed. However, failure to recognise the condition and its serious nature, together with limited awareness of the importance of early referral for treatment, can result in irreversible paralysis. Therefore, it is essential that nurses providing clinical care for these at-risk patients are able to identify early symptoms, and undertake a thorough patient history and examination, educating the patient and their family about the signs and symptoms, which should be reported as soon as they occur. PMID- 18414335 TI - The role of palliative care in supporting patients: a therapeutic community space. AB - The work of palliative day care extends the philosophy and practice of palliative care to patients in the community. It supports the needs of patients who have a diminished ability to fulfil their family and societal roles. The extent to which these aims reflect current practice, however, has not been fully assessed, particularly in Australia. There is also a lack of research on patients' experiences of palliative day care. The key aim of this study was, therefore, to explore patients' experiences of palliative day care in a Western Australian context. Eight patients (two males and six females) with ages ranging from 44 to 82 years were interviewed. Four broad themes emerged from the data, related to patients' experiences: being bounded physically; temporally; and socially; and the role of palliative day care in supporting patients. This study highlighted the potential for palliative day care to provide a therapeutic community space. Implications of these findings for palliative day care are discussed. PMID- 18414336 TI - Legalisation of euthanasia and assisted suicide: a professional's view. PMID- 18414337 TI - Nurses' management of patients with advanced cancer and weight loss: part 2. AB - Weight loss and eating-related disorders are commonly seen in patients with advanced cancer. Using a qualitative approach this study set out to explore how nurses currently manage weight loss and eating difficulties in everyday practice as well as exploring what guides their decision-making with regard to choice of nutritional interventions and support strategies employed. Fourteen semi structured interviews were conducted with nurses from both hospital and community settings. Analysis of the findings revealed that many nurses use an unplanned and reactive style of nutritional management. The findings provide useful insights and potential explanations to an area where, to date, there has been little published research. PMID- 18414338 TI - The benefits and drawbacks of syringe drivers in palliative care. AB - This article will outline the use of continuous subcutaneous infusion pumps, known as syringe drivers, including their benefits and drawbacks in a palliative care context. There have been over 5000 articles published globally describing syringe drivers in the medical and nursing literature within the last decade. Many provide guidance on their use, although much of the data are repetitious, disease or age-group specific, and focused on pragmatic issues to do with clinical application. Several trusts and hospices across the UK are carrying out trials of the recently launched McKinley T34 syringe driver. Therefore, it seems timely to consider their wider use internationally. Globally, practitioners in palliative care are very familiar with their use, although the literature lacks specific guidance and, at times, the information is ambiguous. Having briefly reviewed their benefits, the article considers the limitations of using syringe drivers and comments on some of the lesser known/reported practical and patient focused drawbacks associated with their use. We conclude by considering why, when so much education and training exists to help practitioners use these devices effectively, so many human errors occur. PMID- 18414339 TI - Palliative care professionals' perceptions of nutrition and hydration at the end of life. AB - The provision of medically administered nutrition and hydration (MNH) for the terminally ill patient is a controversial issue and there has been much debate in the literature concerning this sensitive subject. This article reports on a qualitative research study that explores palliative care nurses' and doctors' perceptions and attitudes to patient nutrition and hydration at the end of life. Participants were from an urban and rural palliative care service. Three main discourses were identified: carers' distress at the non-provision of MNH; palliative care doctors' and nurses' position that terminal dehydration lessened the burden of suffering for dying patients; and polarisation between the acute care setting and the palliative care setting. Overlaying these three main discourses are contesting discourses involving cure vs comfort, and acute care vs palliative care. Importantly, the findings of this study reveal that palliative doctors and nurses believe that medically assisted nutrition and hydration at the end stage of life rarely benefits patients, and as long as adequate mouth care is given, patients do not suffer. However, family members do experience emotional distress in dealing with this situation. In caring for dying people, the nurse's and doctor's role is one of education and communication, involving a team approach to manage this difficult issue. PMID- 18414340 TI - [The role of metalloproteinases and their inhibitors in pancreatic cancer]. AB - Pancreatic cancer is characterized by aggressive behavior, poor prognosis, and predicted shortened survival. It is a major cause of cancer death in Europe and North America. Matrix metalloproteinases and their inhibitors play an important role in tumor progression. MMPs are able to degrade basement membrane and extracellular matrix and are associated with tumor progression, including invasion, metastasis, growth, migration, and angiogenesis. Some clinical investigations have demonstrated the role of increased MMP expression in several human malignancies, their levels also correlating with tumor stage, invasiveness, and poor survival of patients with pancreatic cancer. PMID- 18414341 TI - Treatment of atrial fibrillation: is it really going to be a surgical market? PMID- 18414342 TI - Left ventricular reconstruction for severely dilated heart. PMID- 18414343 TI - Fulminant myocarditis treated with percutaneous cardiopulmonary support system (PCPS). AB - PURPOSE: Fulminant myocarditis is characterized by rapid and extensive hemodynamic compromise occurring in a previously healthy patient. The patients sometimes require mechanical circulatory support to maintain systemic perfusion. The purpose of this study was to analyze the clinical course of patients with fulminant myocarditis treated with a percutaneous cardiopulmonary support system (PCPS). PATIENTS AND METHODS: From January 1998 to November 2006, four fulminant myocarditis patients were admitted to the intensive care unit (ICU) in Gunma University Hospital and treated with PCPS to support deteriorating hemodynamics. The mean age of the 4 was 38 +/- 18 (range 14 to 57) years. None of the patients had a past history of heart disease, and the diagnosis of fulminant myocarditis was made with clinical findings and endomyocardial biopsy. Three patients were successfully removed from PCPS; one was not removed and died from cerebral bleeding. Changes in clinical findings, APACHE II scores, and laboratory data were analyzed in the 3 survivors and 1 nonsurvivor. RESULTS: Intra-aortic balloon pumping (IABP) was used in all 4 patients. The duration of PCPS support was 141, 228, and 266 h in the survivors and 330 h in the nonsurvivors. The interval between the occurrence of clinical symptoms such as fever and general fatigue and the induction of PCPS in the nonsurvivor was shorter (2 days) than in the survivors (4-6 days). Cardiac troponin I (cTnI) and creatine phosphokinase (CPK) MB levels were significantly higher in the nonsurvivor compared with those in the survivors. Left ventricular ejection fraction (LVEF) gradually improved, and PCPS flow was decreased at around 120 h after PCPS start in the survivors; however, these improvements did not occur in the nonsurvivor. CONCLUSION: PCPS was induced in 4 fulminant myocarditis patients and successfully removed from 3 after long term PCPS (>5 days). The maintenance of hemodynamics, especially in the acute phase of fulminant myocarditis, is important because the possibility of circulatory recovery is relatively high compared with those having severe cardiac failure resulting from other causes. The prognosis might be poor if the interval between the occurrence of clinical symptoms and PCPS deployment is short. PMID- 18414344 TI - Comparison of the effects of aortic valve replacement using 19-mm Carpentier Edwards Perimount bioprosthesis and 19-mm Medtronic Mosaic bioprosthesis. AB - OBJECTIVE: Short (< or =3 months)- and middle (> or =4 months)-term results of aortic valve replacement (AVR) using 19-mm Carpentier-Edwards Perimount (CEP) bioprosthetic valves and 19-mm Medtronic Mosaic (MM) bioprosthetic valves in patients with small aortic annulus were compared. PATIENTS AND METHODS: At our facility, AVR was performed using bioprostheses in 110 patients from April 1999 to March 2006. Of these patients, 40 were treated using 19-mm CEP (Group C), and 9 using 19-mm MM (Group M). Evaluation by inquiry, physical examination, and echocardiography was performed before, a short term after, and a middle term after surgery, and the effects of AVR were compared. RESULTS: The New York Heart Association (NYHA) functional class grade showed improvements in both groups. The aortic valve peak pressure gradient was 29.8 +/- 10.1 mmHg in Group C and 53.8 +/ 17.3 mmHg in Group M, being higher in Group M, a middle term after surgery. However, the left ventricular mass index (LVMI) showed improvements in both groups compared with the values before surgery, and the left ventricular ejection fraction (LVEF) was maintained. During the middle term after surgery, the frequency of cardiac events showed no significant difference between the two groups. CONCLUSIONS: In the patients treated with 19-mm MM, the aortic valve peak pressure gradient was higher than in those treated with 19-mm CEP, but acceptable improvements in the LVMI, maintenance of the LVEF, and avoidance of cardiac events were observed in both groups. PMID- 18414345 TI - Recommendation for appropriate use of GRF glue in the operation for acute aortic dissection. AB - BACKGROUND: Because an excessive use of activator (formaldehyde + glutaraldehyde) is supposed to be responsible for later adverse events after the use of gelatin resorcin formalin (GRF) glue in surgery for acute aortic dissection, we have tried to use a minimum dose of activator when the GRF glue was applied. We compare our midterm surgical results for acute aortic dissection with and without the use of GRF glue. METHODS: Forty-nine consecutive operated cases with Stanford type A acute aortic dissection within 48 h from onset from 1992 to 2005 were retrospectively analyzed. GRF glue was used in 21 cases (18 proximal and 14 distal anastomosis sites) since 1995 with outer felt reinforcement (GRF group). RESULTS: There was no operative deaths. In-hospital mortality was 4.8% in the GRF group and 7.1% in the control group (P = 0.7308). Intraoperative blood loss and transfusion requirements were similar between groups. The patency of the distal false lumen after the operation (57% vs. 55%, P = 0.8855), the 3-year survival estimate (82% +/- 10% vs. 92% +/- 6%, P = 0.4219), and the 3-year actuarial freedom from a reoperation of 92 anastomoses (97% +/- 3% vs. 100%, P = 0.4986) were similar between the GRF group and the control group, respectively. A multivariate Cox's proportional hazard model identified no significant predictor for midterm death or reoperation. CONCLUSIONS: The use of GRF glue for type A acute aortic dissection seems as clinically safe as other options with regard to midterm death or reoperation when applied appropriately with felt reinforcement. PMID- 18414346 TI - The outcome of patients undergoing abdominal aortic aneurysm repair with and without cardiac disease. AB - PURPOSE: We report the outcome of abdominal aortic aneurysm (AAA) repair with and without cardiac disease. In patients with cardiac disease, we compared staged procedures and simultaneous operations. MATERIAL AND METHODS: A total of 217 patients underwent scheduled AAA repair from 1994 to 2005. When the AAA with cardiac disease was 5.0 cm or less in diameter, we performed a staged procedure. When it was more than 5.0 cm in diameter, we performed a simultaneous operation. Forty-two patients underwent simultaneous operations, and 15 had staged procedures. RESULTS: No occurrence of major cardiac complication was detected in any patient postoperatively. There were no hospital deaths in either the no cardiac-disease group or the staged-procedure group. There were two hospital deaths in the simultaneous group. The estimated actuarial 10-year survival rates were 58.9 +/- 9.9% and 68.2 +/- 1.5% for the no-cardiac-disease group and the with-cardiac-disease group, respectively (P = 0.85). The 10-year survival rates were 71.2 +/- 1.6% and 74.8 +/- 1.3% for the one-stage-operation group and the staged-operation group, respectively (P = 0.35). There was no significant difference with regard to hospital deaths and late mortality rate among the groups. CONCLUSION: The simultaneous operation method is a useful technique in patients having both a large AAA and cardiac disease. PMID- 18414347 TI - Left sleeve Basal segmentectomy for broncholithiasis. AB - Broncholithiasis is an uncommon pulmonary problem that may present with life threatening complications. We report one case of broncholithiasis. A 49-year-old female presented with hemoptysis. Chest X-ray and computed tomography (CT) showed left interlobar lymph node calcification near the interlobar pulmonary artery, and calcification continued into the basal bronchus. Bronchoscopy demonstrated pedunculated granulation tissue in the left B8 bronchus obstructing the lumen. We did not recommend endobronchial removal because of the risk of bleeding, so we proposed surgical treatment. We performed left basal segmentectomy associated with bronchoplasty to preserve pulmonary function. It is important to gain proximal control of the pulmonary artery before dissection of its branches and to approach the pulmonary artery from the periphery to avoid massive intraoperative bleeding. The intrabroncholuminal stone was composed of 61% calcium carbonate and 39% calcium phosphate. The postoperative course was not eventful, and the bronchoscopical findings confirmed a good surgical outcome. PMID- 18414348 TI - Surgical treatment of endobronchial leiomyosarcoma with right main bronchus total obstruction: a case report. AB - Endobronchial leiomyosarcoma is an unusual tumor of the respiratory tract. Clinically, patients may present with intermittent coughing, chest pain, dyspnea, hemoptysis, and fever until late in the course of the disease because of total obstruction of the main airway. In this paper, we report the case of a 51-year old male with endobronchial leiomyosarcoma who presented with acute respiratory distress as a result of total obstruction of the right main bronchus and suffocation after massive hemoptysis. After intraoperative bronchoscopic assessment and bronchotomy, an elongated endobronchial tumor was found that arose from the right middle lobe (RML) bronchus with intraluminal extension upward into the right main bronchus. He underwent RML and right lower lobe (RLL) bilobectomy and had a rapid and uneventful recovery. PMID- 18414349 TI - Pulmonary typical carcinoid tumor and liver metastasis with hypermetabolism on 18 fluorodeoxyglucose PET: a case report. AB - Pulmonary carcinoid tumors are generally hypometabolic on 18-fluorodeoxyglucose (FDG)-positron emission tomography (PET). We experienced a case of pulmonary typical carcinoid that showed rapid growth and high FDG uptake at the primary site and liver metastasis. A 56-year-old man with hemosputum had a medical examination by his family physician. A roentgenogram and computed tomography of the chest showed a solitary solid mass on the right lower lung field. However, he had not been shown an abnormal shadow on a roentgenogram taken 8 months earlier. He had undergone fiber-optic bronchoscopy, but the cytological diagnosis showed no evidence of malignancy. After that, FDG-PET was examined and revealed hot spots in the pulmonary tumor and liver mass. A standard uptake value of this pulmonary tumor was 32.9 mg/mL, and that of the liver mass was almost the same value of pulmonary lesion. He had undergone a right lower lobectomy diagnosed as a typical carcinoid. Thereafter he underwent partial resection of he liver mass, and the histology was metastasis from pulmonary carcinoid. We first reported a typical pulmonary carcinoid that showed high FDG uptake at the primary site and liver metastasis. PMID- 18414350 TI - Functional large parathyroid carcinoma extending into the superior mediastinum. AB - Mediastinal parathyroid carcinoma is very rare regardless of whether it is functioning, and it is sometimes problematic in both diagnosis and treatment. We herein report a case of primary hyperparathyroidism (PHP) associated with large parathyroid carcinoma extending into the superior mediastinum successfully removed by a transcervical approach, with a review of recent literature. A 61 year-old male with a six-year history of PHP and urinary stone disease presented with an abnormal mediastinal mass in a chest X-ray. His serum level of intact parathyroid hormone (PTH) was markedly elevated to 1,220.0 pg/mL (normal range: 10.0-65.0 pg/mL), though calcium and phosphorus levels were within normal limits. Chest computed tomography demonstrated a large mass in the superior mediastinum, displacing the trachea to the right and reaching the aortic arch, posteriorly adjacent to the thoracic vertebra. Transcervical extirpation of the tumor was performed. The resected specimen measured 50x85x38 mm and weighed 56.8 g. Histopathological examination revealed a capsular invasion of the tumor cells, and a diagnosis of parathyroid carcinoma was made. On the 8th postoperative day, the patient was discharged uneventfully with a decreased serum level of PTH (59.0 pg/mL), and no recurrent disease or exacerbation of hyperthyroidism was observed 36 months after surgery. PMID- 18414351 TI - Surgical management of Aspergillus colonization associated with lung hydatid disease. AB - Colonization with Aspergillus sp. usually occurs in previously formed lung cavities. Cystectomy is a widely used surgical technique for hydatid lung disease that can also leave residual cavities and potentially result in aspergilloma. We present two cases of this rare entity and a case with Aspergillus sp. colonization of an existing ruptured hydatid cyst. PMID- 18414352 TI - Long-term clinical outcome after extended thymectomy combined postoperative high dose steroid therapy for juvenile myasthenia gravis. AB - Myasthenia gravis (MG) is considered to be an autoimmune disorder of neuromuscular transmission and is rare in childhood. We report 3 juvenile MG (JMG) cases of extended thymectomy (ETMX) combined postoperative high-dose steroid therapy. All patients developed MG symptoms under the age of 14 years and were given cholinergic drugs and had generalized MG: the Myasthenia Gravis Foundation of America classification II was present in 1 case and classification III was in 2. All patients were taking pyridostigmine before surgery; none was treated with prednisone preoperatively. All patients performed the ETMX combined postoperative high-dose steroid therapy. Muscle weakness of extremities and bulbar symptoms improved in all patients, but not all exhibited an unstable phase in their clinical course as a result of combined therapy. There was no postoperative morbidity or mortality. All patients had follicular lymphoid hyperplasia without thymoma. Follow-up for more than 5 years has shown one to be in complete remission and the others to have improved symptoms. Although our results are inconclusive because we used only a few JMG patients, the ETMX combined postoperative high-dose steroid therapy appeared to provide a better chance of remission or control of symptoms. PMID- 18414353 TI - Successful replacement of aortic valve using a stentless porcine valve in a patient with myelodysplastic syndrome. AB - Open heart surgery in patients with myelodysplastic syndrome (MDS) increases the risk of bleeding and infection. We document a 78-year-old female with severe aortic valve stenosis and MDS who underwent aortic valve replacement with stentless porcine prosthesis. Perioperatively, a transfusion of red blood cells (RBCs) and an infusion of granulocyte colony-stimulating factor (G-CSF) were needed for serious erythrocytopenia and leukocytopenia. We did not need a large amount of RBC transfusions in the postoperative course, and no infection was seen postoperatively. PMID- 18414354 TI - Microembolization from an abdominal aortic aneurysm after thoracic aortic replacement. AB - A 66-year-old man with thoracic and abdominal aortic aneurysm suffered from microembolism in the lower extremities after total arch replacement. He presented with livedo reticularis with palpable peripheral pulses, and the serum creatinine kinase level elevated up to 7,695. The abdominal aortic aneurysm, but not the thoracic aorta, was the origin of this complication. The morphological change of thrombus in the abdominal aorta detected by ultrasonography was the key to the diagnosis. Graft replacement of the abdominal aorta finally resolved his problem. PMID- 18414355 TI - Nonocclusive mesenteric ischemia after aortic surgery in a hemodialysis patient. AB - We report a case of nonocclusive mesenteric ischemia (NOMI), which developed after grafting for a descending thoracic aortic aneurysm in a hemodialysis patient. On postoperative day 5, acute increases in serum enzyme levels developed. Emergency angiography revealed severe vasoconstriction in the superior mesenteric artery (SMA) and other splanchnic arteries. Therefore an infusion of papaverine hydrochloride was started into the SMA. Although serum enzyme levels decreased, metabolic acidosis occurred the next day. An emergency laparotomy revealed segmental diffuse necrotic small intestine and colon. Despite a resection of the small intestine and sigmoid colon, the patient died of septic shock several days later. NOMI is uncommon, but it is a catastrophic event that can occur after cardiovascular surgery. If intestinal gangrene is suspected, prompt mesenteric angiography and vasodilator therapy followed by exploratory laparotomy should be performed without delay. PMID- 18414356 TI - PET imaging in endocrine tumours. AB - The role of PET in the assessment of endocrine tumours has been, until recently, restricted to the use of (18)F-fluoro-deoxy-D-glucose ((18)F-FDG). Being a marker of metabolically active lesions that show high grading and low differentiation, FDG is not ideal for this purpose since the majority of endocrine tumours are slow growing and highly differentiated. It is however useful when dedifferentiation takes place and provides excellent prognostic information. A number of hormone precursors and amino acids are labelled with (11)C and used successfully in the management of parathyroid, adrenal and pituitary tumours. However, the short half-life of (11)C radiopharmaceuticals restricts their use to centres with access to an on-site cyclotron, while the high cost of production may limit their use to research purposes. A promising new positron-emission tomography (PET) tracer is Gallium-68 obtained by elution from a long shelf-life generator that makes it economic and cyclotron-independent. Its short half-life and flexible labelling ability to a wide range of peptides and antibodies makes it ideal for PET imaging. In addition to imaging GEP-NETs and phaeochromocytoma, it has the potential to be used in a wider range of endocrine tumours. PMID- 18414357 TI - Treating bacteremia in critical patients: a long course of combined antimicrobial therapy or a short course with a single antibiotic? PMID- 18414358 TI - Italy versus the United Kingdom: differing styles for treating bacteremia in the critically ill patient, but who's right? AB - BACKGROUND: As no prospective, randomized, clinical trials (PRCTs) exist to inform decisions about antibiotic therapy for bacteraemia, ICUs have likely developed their own strategies. A multi-centre, multi-national questionnaire survey was used to assess variations in antibiotic strategy between ICUs in the United Kingdom (UK) and Italy. METHODS: In January 2001, a detailed questionnaire regarding antibiotic policy was sent to international (e.g., European Society of Intensive Care Medicine, ESICM) and national societies (Gruppo Italiano per la Valutazione degli Interventi in Terapia Intensiva, GIVITI; Australia and New Zealand Intensive Care-Critical Trial Group, ANZIC-CTG; United Kingdom-Intensive Care Society, UK-ICS). RESULTS: Out of the 254 participating ICUs, 81 were from Italy and 32 were from the UK. In the UK, ICUs preferred to use a shorter course of mono-therapy compared to Italian ICUs when treating, a) primary bacteremia [7 (5-7) vs 10 (7-14), P<0.001]; b) UTI [5 (4-7) vs 8 (7-10), P<0.001]; c) Staphylococcus aureus [7 (5-12) vs 10 (7-15), P<0.001]; and d) Gram-negative bacteraemia associated with lines [6 (5-7) vs 10 (7-15), P<0.001], pneumonia [7 (5-7) vs 14 (10-15), P<0.001] and peritonitis [7 (5-9) vs 15 (10-15), P<0.000]. These differences in treatment strategy were likely correlated with the median inter-quartile range (IQR) of weekly input from a microbiologist or infectious disease specialist, which was more frequent in the UK compared to Italy [5 (2-6) vs 0 (0-1), P<0.001]. CONCLUSION: Possible influences accounting for the variations found between Italy and the UK are: 1) differences in interactions with microbiologists or infectious disease specialists, 2) historical/educational dogma, and 3) antibiotic resistance patterns. Further studies are necessary to determine the optimal approach to treating bacteremia. PMID- 18414359 TI - Effect of nitrous oxide on desflurane MACBAR at two target-controlled concentrations of remifentanil. AB - BACKGROUND: The aim of this prospective, randomized, double-blind study was to determine the effect of nitrous oxide on the desflurane requirement for blunting sympathetic response following surgical incision (MACBAR) when desflurane was combined with two different target-controlled concentrations of remifentanil (1 and 3 ng/mL). METHODS: A total of 103 patients, aged 20-50 years, ASA physical status I, scheduled to undergo general anesthesia for elective abdominal surgery, were randomly allocated to receive anesthesia with desflurane alone (Group A, n = 53), or with the addition of 60% nitrous oxide (Group N, n = 50). Patients of both groups were further assigned to receive a target-controlled plasma concentration of 1 ng/mL (Group A1, n = 27; Group N1, n = 26) or 3 ng/mL remifentanil (Group A3, n = 26, Group N3, n = 24). Sympathetic responses to surgical incision were determined after a 20 min period of constant end-tidal desflurane and target-controlled remifentanil concentrations. Predetermined end tidal desflurane concentrations and the MACBAR for each group were determined using an up-and-down sequential allocation technique. RESULTS: A total of 98 patients completed the study. The MACBAR of desflurane was 5.2% (95% confidence interval [CI95: 4.9-5.5%]) in Group A1 and 2.7% (CI95: 2.6-2.8%) in Group N1 (P<0.001), while in Groups A3 and N3 the MACBAR of desflurane was 2.2% (CI95:2 2.4%) and 2% (CI95:1.9-2.2%), respectively (P<0.01). When considering a minimum anesthetic concentration (MAC) value with a contribution of 60% nitrous oxide (0.55 MAC) in this population, the combined MACBAR values (expressed as multiples of the MAC) were 1.9 MAC for group A1, 1 MAC for group N1, 0.8 MAC for group A3, and 0.7 MAC for group N3. CONCLUSION: The addition of 60% nitrous oxide reduces the MACBAR of desflurane by 52% when using a remifentanil concentration of 1 ng/mL, and reduces the MACBAR by 10% when using a remifentanil concentration of 3 ng/mL. PMID- 18414360 TI - Efficacy of propofol compared to midazolam as an intravenous premedication agent. AB - BACKGROUND: It is common practice to administer a premedication to patients about to undergo anesthesia. This study compared the effects of a small intravenous bolus of propofol versus midazolam administered as a premedication. METHODS: In this prospective, randomized, double-blind, placebo-controlled study, 60 healthy adult subjects scheduled for general anesthesia were given midazolam (0.04 mg/kg), propofol (0.4 mg/kg) or saline intravenously in the anesthesia waiting room. Before administration and three times at 2.5 min intervals subsequently, blood pressure, heart rate, oxygen saturation and scores for anxiety and dizziness were recorded. Recall of words and images shown to the patient 5 min after drug administration was tested 10-30 min after recovery from anesthesia. RESULTS: Both propofol and midazolam helped relieve anxiety and lowered blood pressure (P<0.001, compared to baseline), but both were associated with greater dizziness scores (P<0.001, compared to placebo). Compared to placebo and propofol, midazolam was associated with more frequent respiratory depression (P<0.05) and significant impairment of anterograde explicit memory (P<0.05). CONCLUSION: Propofol may be an economical and safe alternative to midazolam for i.v. premedication. PMID- 18414361 TI - Management of sepsis. AB - An early diagnosis of sepsis prior to the onset of clinical decline is of particular interest to health practitioners because this information increases the possibilities for early and specific treatment of this life threatening condition. In comparison to acute myocardial infarction or ischemic stroke, the time to initiate therapy is thought to be crucial and the major determining factor for surviving sepsis. The treatment of severe sepsis and septic shock consists of source control, early antimicrobial therapy, and supportive and adjunctive therapies. For supportive therapy, an adequate volume loading is the most important step in the treatment of patients with sepsis. This step is performed in order to restore and maintain oxygen transport and tissue oxygenation. Therefore, the supportive treatment should focus on adequate volume resuscitation and appropriate use of inotropes and vasopressors. Within the first 24 h after the initial sepsis-induced organ failure, adjunctive therapies can help to decrease mortality in patients suffering from severe sepsis and septic shock. Ongoing research continues to provide new information on the management of sepsis. However, implementing new medical advances in the management of sepsis into daily clinical intensive care remains a major hurdle. High quality management tools are necessary to bring evidence-based therapy to the bedside. With respect to recently published studies, the importance of the time taken to improve the outcome of sepsis can not be overemphasized. PMID- 18414362 TI - Intracranial pressure monitoring for traumatic brain injury: available evidence and clinical implications. AB - Following traumatic brain injury, uncontrollable intracranial hypertension remains the most frequent cause of death. Despite general agreement on the deleterious effects of elevated intracranial pressure (ICP), however, the evidence supporting the use of ICP monitoring has recently been questioned. The aim of this review was to evaluate the pros and cons of ICP monitoring and to discuss the hypothetical desirability and feasibility of a trial testing the benefits of ICP monitoring. PMID- 18414363 TI - Rhabdomyolysis and respiratory failure: rare presentation of carnitine palmityl transferase II deficiency. AB - Carnitine palmityl-transferase (CPT) II deficiency is a rare disorder of the fatty acid beta-oxidation cycle. CPT II deficiency can be associated with rhabdomyolysis in particular conditions that increase the requirement for fatty acid oxidation, such as low-carbohydrate and high-fat diet, fasting, exposure to excessive cold, lack of sleep and prolonged exercise. The best known CPT II deficiency is the muscular form with episodic muscle necrosis and paroxysmal myoglobinuria after prolonged exercise. We report a case of a four-year-old male child, who, after one day of hyperthermia and fasting, developed a massive rhabdomyolysis beginning with acute respiratory failure and later complicated by acute renal failure. Appropriate management in Pediatric Intensive Care Unit (PICU) (mechanical ventilatory support, fluid supply combined with mannitol and bicarbonate infusions, administration of acetaminophen and antibiotics, and continuous venovenous haemofiltration) brought about complete resolution with an excellent outcome. Biochemical investigation of muscle biopsy and genetic analysis showed a deficiency of CPT II. The onset of CPT II deficiency with respiratory failure is extremely rare, but a correct and early diagnosis of rhabdomyolysis is the key to successful treatment. A metabolic myopathy such as CPT II deficiency should be suspected in children affected by rhabdomyolysis if trauma, crash, infections, drugs or extreme exertion can be excluded. PMID- 18414364 TI - Unanticipated discovery of vascular anomalies by hemodialysis catheter placement. Case report. AB - This case report describes an unexpected finding post hemodialysis catheter placement in a child scheduled for living-relation renal transplant. Moreover, the unusual appearance of the catheter on a chest X-ray prompted further investigation of the patient's vascular anatomy, resulting in the discovery of an aberrant iliac artery course that significantly affected the surgical approach to this planned procedure. PMID- 18414365 TI - The history of anesthesia for thoracic surgery: some remarks. PMID- 18414366 TI - Effects of remifentanil on propofol requirements for loss of consciousness in target-controlled infusion. PMID- 18414367 TI - A prospective, randomized, double-blind comparison between parecoxib and ketorolac for early postoperative analgesia following nasal surgery. AB - BACKGROUND: The aim of this prospective, randomized, double-blind study was to compare the efficacy of parecoxibfor postoperative analgesia after endoscopic turbinate and sinus surgery, with the non-selective non-steroid anti-inflammatory drug (NSAID), ketorolac. METHODS: A total of 50 patients with an ASA physical status I-II, receiving functional endoscopic sinus surgery (FESS) and endoscopic turbinectomy after local infiltration with 1% mepivacaine, were randomly assigned to receive intravenous administration of either 40 mg parecoxib (N.=25) or 30 mg ketorolac (N.=25), 15 min before the discontinuation of anaesthesia and then every 8 h postoperatively. A blinded observer recorded the incidence and severity of pain upon admission to the postanesthesia care unit (PACU), as well as 10, 20, and 30 min after PACU admission. Thereafter, observations continued every 1 h for the first 6 h, and then 12 h and 24 h after surgery. RESULTS: The area under the curve of the visual analogue scale (AUCVAS) calculated during the study period was 635 (26-1 413) in the Parecoxib group and 669 (28-1 901) in the Ketorolac group (P=0.54). Rescue morphine analgesia was required by 12 patients (48%) in the Parecoxib group and 11 patients (44%) in the Ketorolac group (P<0.05); while mean morphine consumption was 5 +/- 2.5 mg and 5 +/- 2.0 mg in Ketorolac and Parecoxib groups, respectively (P<0.05). No differences in the incidence of side effects were recorded between the two groups. Patient satisfaction was similarly high in both groups, and all patients were discharged uneventfully 24 h after surgery. CONCLUSION: In patients undergoing endoscopic nasal surgery and local infiltration with 1% mepivacaine, parecoxib administered before discontinuing general anesthesia is as effective in treating early postoperative pain as ketorolac. PMID- 18414368 TI - Sufentanil-propofol vs remifentanil-propofol during total intravenous anesthesia for neurosurgery. A multicentre study. AB - BACKGROUND: In a randomised, prospective multi-centre study, we compared the intraoperative and postoperative effects of two opioids: sufentanil and remifentanil, in combination with propofol in two groups of patients undergoing neurosurgery. METHODS: After Local Ethics Committee approval and informed consent obtaining, 69 patients undergoing neurosurgery for supratentorial tumours, between 18 and 75 years of age were randomised to receive either sufentanil or remifentanil in combination with propofol. Intraoperative and postoperative haemodynamic variables, recovery times (time to eye opening and to extubation), the incidence of postoperative respiratory depression, pain, nausea and vomiting were also evaluated. The Short Orientation-Memory-Concentration Test was used to evaluate cognitive function at 15, 45 and 180 min after emergence from anesthesia. RESULTS: There were no significant differences between the groups in the duration of surgery and anesthesia, mean arterial pressure, heart rate, time to eye opening or extubation. The incidence of vomiting, respiratory depression and shivering was similar in both groups. Postoperative pain requiring supplemental analgesics was significantly lower in the sufentanil group (P<0.05). Although there were no significant differences between the groups in postoperative behavioural examinations by Rancho Los Amigos Test, patients anesthetised with sufentanil had significantly better Short Orientation-Memory Concentration Test values at 15 and 180 min postoperatively (P<0.05). CONCLUSION. We conclude that remifentanil and sufentanil are suitable adjunct to propofol for total intravenous anesthesia (TIVA). Patients receiving sufentanil have reduced analgesic requirements and better cognitive function postoperatively than those who received remifentanil. PMID- 18414369 TI - The use of helmets to deliver non-invasive continuous positive airway pressure in hypoxemic acute respiratory failure. AB - Non-invasive continuous positive airway pressure (CPAP) is a useful tool for managing patients with acute respiratory failure. The head helmet is a relatively novel interface that is as effective as the traditionally employed face-mask in delivering CPAP and can possibly be characterized as better for the patient's tolerance and, consequently, a longer duration of treatment. This review focuses on the main properties of the helmet and the issues related to its use, as shown by the physiological and bench studies. Clinical experience, both personal and reported in the literature, for the treatment of both cardiogenic and non cardiogenic pulmonary edema is reviewed as well. PMID- 18414370 TI - Levobupivacaine plus fentanyl versus racemic bupivacaine plus fentanyl in epidural anaesthesia for lower limb surgery. AB - BACKGROUND: To compare the anaesthetic epidural effects of levobupivacaine plus fentanyl versus bupivacaine plus fentanyl in patients undergoing lower limb surgery. METHODS: A single blind, randomised, prospective, multicentre study was designed to compare both therapies. The study was conducted in 10 tertiary hospitals. A total of 96 patients who were ASA I or II, who required at least a 24-hour-stay in the hospital and who were subjected to surgery of lower limbs with epidural anaesthesia were enrolled in this study. Treatments were administered at a dosage of 1.2 ml per metamera,including a test dose (3 mL) and the dose of fentanyl (100 mg). Patients were then randomly allocated to receive either Levobupivacaine (n = 49) or bupivacaine (N.= 47). The primary endpoint was sensory blockade (SB) duration. Secondary evaluations included motor blockade (MB), post-surgery analgesic medication usage, safety and the investigator global evaluation. RESULTS: SB duration was similar for both interventions: 195 min (165 205) in the bupivacaine group versus 170 min (140-185) in the levobupivicaine group (log-rank test, P=0.884). However, the lack of MB as evaluated by the modified Bromage scale was significantly higher in the levobupivacaine group than in the bupivacaine group (39% vs 13%, P=0.017). Although no significant differences in MB duration were observed between the groups, a trend was observed in the levobupivacaine group, which had a lesser MB (P=0.093). Investigator satisfaction was high and was assessed to a similar extent for both interventions. Forty-one adverse events were detected in 28 patients, with no differences between groups: 15 (33%) with bupivacaine and 13 (27%) with levobupivacaine, P=0.516. CONCLUSION: Although both interventions showed similar anaesthetic effects, a higher proportion of patients receiving levobupivacaine lacked MB. PMID- 18414371 TI - The ability of PiCCO versus LiDCO variables to detect changes in cardiac index: a prospective clinical study. AB - BACKGROUND: Both PiCCO and LiDCO can provide dynamic preload parameters, pulse pressure variation (PPV) and stroke volume variation (SVV). The PiCCO device also provides a measure of intrathoracic blood volume index (ITBVI). We investigated the agreement between SVV and PPV, as well as the reliability of LiDCO- and PiCCO measured SVV, PPV and ITBVI, in detecting fluid responsiveness before and after fluid challenge (FC). METHODS: We performed a prospective clinical study in University Hospital ICU. Nine adult ICU patients with cardiovascular instability were enrolled in the study. All patients were sedated and mechanically ventilated with intermittent positive pressure ventilation. The PiCCO and LiDCO systems were both connected to each patient. The PiCCO pulse waveform system was joined by a 5 French (Fr) thermistor-tipped arterial catheter inserted into the femoral artery. LiDCO measurements were performed through radial artery pulse contour analysis. Fluid challenge was performed using a rapid infusion of 7 mL/kg of 6% hydroxyethylstarch over 30 min. RESULTS: Measurements of CI, ITBVI, SVV, and PVV were made using both techniques before and after FC. Pre-FC cardiac index (CI) measurements were similar with both devices, although the reading was higher after FC with the PiCCO device (P<0.001). The correlation coefficient between PiCCO-CI and LiDCO-CI was 0.85 (95% CI: 0.69 to 0.93; P<0.001); for P-PPV and L PPV, it was 0.74 (95% CI: 0.49 to 0.88; P<0.001). Only ITBV had a significant correlation with LiDCO-CI or PiCCO-CI. CONCLUSION: We found a narrow bias but less accurate precision in cardiac index values measured by a radial artery-site LiDCO catheter and a femoral artery-site PiCCO catheter, with poor agreement between radial and femoral-derived SVV and PPV measurements. ITBVI proved to be the best predictor of fluid responsiveness. The SVV does not seem to be reliable for preload optimization in ICU patients. PMID- 18414372 TI - Helminth control using local resources in smallholder production systems of Asia. AB - Practical helminth control in smallholder systems needs to match technical options with local knowledge, locally available animal and feed resources and the needs of both producers and consumers. Despite extensive research over many decades the uptake of new technology in these systems has been slow and limited to few farmers with access to good technical support. Investment by small holders is constrained by lack of regular market signals for livestock and livestock products. Examples of effective helminth control in cattle, buffalo, sheep, goat and pig systems show that effective control is possible using local resources and knowledge. Livestock systems in Asia are changing rapidly through industrialisation of production and consolidation of supply chains and retail markets, but smallholder systems, which are resilient and multipurpose, will remain important in rural areas and in peri-urban environments. They also provide a significant pathway for the poor to build assets and generate income. Helminth control in these systems will always vary greatly between farmers and systems and will need to be relatively simple and tailored to locally available resources. The public sector will continue to provide advice on appropriate genetics, to regulate drug importation, use and quality, and ensure novel helminth control options are investigated for local application and promotion to livestock producers. The private sector has the complementary role to develop clear market signals for livestock and livestock products, and make anthelmintics available in appropriate packages. Improved helminth control has the potential to increase the profitability and sustainability of all components of the livestock sector. PMID- 18414373 TI - Targeted treatment strategies for sustainable worm control in small ruminants. AB - Sustainable worm control strategies are based largely on ensuring that a source of worms not exposed to anthelmintics ("in refugia") remains after treatments are given, so that resistant worms do not become a dominant part of the total population. In environments with seasonally poor survival of worm larvae on pasture, this may require withholding treatments from a proportion of animals when the whole group would normally be treated. The "targeted treatment" approach involves using anthelmintics on an individual animal basis according to indications of parasitic effects, regardless of parasite burdens. For Haemonchus contortus, the FAMACHA system, based on the easily-visualised index of anaemia, has proved effective provided that labour is available for frequent inspections. For non-haematophagous nematodes, recent research indicates the potential of production parameters such as body weight change (sheep) and milk yield (dairy goats), providing that parasitic effects can be differentiated from nutritional and other factors. Continuing investigations are necessary to indicate the most appropriate indices for different situations, so that the refugia effect is maximized for the least risk of disease and production loss. Of prime importance, targeted treatment strategies must be practical to implement if they are to achieve widespread adoption. PMID- 18414374 TI - Future perspectives for the implementation of genetic markers for parasite resistance in sheep. PMID- 18414375 TI - Managing refugia. PMID- 18414376 TI - Challenges to the development of new tests for diagnosis of infection and prediction of resistance of sheep to gastrointestinal nematodes. AB - Strategies for control of gastrointestinal nematode (GIN) infections in sheep require information on the severity of infection and species (or genus) of parasite present. Tests for diagnosis of GIN fall into 3 classes. Current tests measure either: 1) the presence of eggs, worm antigens or worms themselves; 2) components of host immunity (e.g. antibodies, eosinophils, other immune mediators); or 3) components of host pathology (e.g. wool growth, body growth, appetite, blood loss, digestive enzymes, anaemia, hypoproteinaemia, odours). To offer advantages over current diagnostic methods, new tests need to be more informative, more accurate, more timely, cheaper, technically easier, or suitable for use in the field. To improve sheep management, a new test needs to determine severity or predict the onset and severity of infection. This is a technically more difficult challenge than qualitative diagnosis of the presence of infection and creates a substantial obstacle to the development of new diagnostic methods. Estimation of the performance characteristics of a test including its sensitivity, specificity and predictive value is important before widespread adoption. This paper reviews current diagnostic tests for GIN, and opportunities for new tests that aid management of infections or that inform the estimated breeding value of animals for use in programs that breed sheep for resistance to GIN. Gene marker and biomarker tests for resistance to GIN infection or disease will require validation in the population in which they are to be used and may require revalidation as the genetic background of the population changes over time. Estimation of the specificity, sensitivity, and predictive value of gene markers and biomarkers for GIN infection may help inform the value of these markers as selection criteria for use in breeding programs. PMID- 18414377 TI - Recent vaccine related studies with economically important gastrointestinal nematode parasites of ruminants. AB - Although several native antigens of sufficient efficacy to be commercially useful have now been discovered for Haemonchus and Ostertagia, all will have to be synthesised artificially to be economically viable. Despite numerous attempts, recombinant DNA technology has not yet yielded the solution, but the effort continues, spurred on by the successes with cestodes and certain ticks. Teladorsagia and Trichostrongylus lag behind in the vaccine research stakes: here, the first and probably most difficult step of identifying a reliably protective native antigen extract does not seem to have been achieved yet. It may be necessary to stimulate elements of the mucosal response to induce protection, a subject still in its infancy as far as ruminants are concerned. Several laboratories have started to work on antigen delivery methods with this in mind. It is easy to understand why intestinal antigens protect against blood feeding Haemonchus rather than mucosal browsers, but quite why conventional immunisation works for Ostertagia, but not apparently for Teladorsagia or Trichostrongylus remains a mystery. PMID- 18414378 TI - Identification and validation of bioactive plants for the control of gastrointestinal nematodes in small ruminants. AB - The search for bioactive plants which can be used as non-conventional anthelmintics (AHs) has received considerable attention in recent times because of the increasing, worldwide development of resistance to chemical anthelmintics in worm populations. However, scientific evidence to validate the use of plants remains limited. The criteria required for the scientific validation of phytomedicine and nutraceuticals are similar to those expected for chemical anthelmintics: definition and quality, efficacy, identification of side-effects. However, the methods of investigation need to take into account i) the variability in bioactive compounds in the natural resources, ii) the mode of distribution to animals and iii) the potential anti nutritional and side-effects of some bioactive compounds and the possible occurrence of regulative mechanisms in the hosts. These points are critically discussed, in particular by reference to the example of tannin rich plants with anthelmintic properties from both temperate and tropical regions. PMID- 18414379 TI - Novel consequences of voltage-dependence to G-protein-coupled P2Y1 receptors. AB - BACKGROUND AND PURPOSE: Emerging evidence suggests that activation of G-protein coupled receptors (GPCRs) can be directly regulated by membrane voltage. However, the physiological and pharmacological relevance of this effect remains unclear. We have further examined this phenomenon for P2Y1 receptors in the non-excitable megakaryocyte using a range of agonists and antagonists. EXPERIMENTAL APPROACH: Simultaneous whole-cell patch clamp and fura-2 fluorescence recordings of rat megakaryocytes, which lack voltage-gated Ca2+ influx, were used to examine the voltage-dependence of P2Y1 receptor-evoked IP3-dependent Ca2+ mobilization. RESULTS: Depolarization transiently and repeatedly enhanced P2Y1 receptor-evoked Ca2+ mobilization across a wide concentration range of both weak, partial and full, potent agonists. Moreover, the amplitude of the depolarization-evoked [Ca2+]i increase displayed an inverse relationship with agonist concentration, such that the greatest potentiating effect of voltage was observed at near threshold levels of agonist. Unexpectedly, depolarization also stimulated an [Ca2+]i increase in the absence of agonist during exposure to the competitive antagonists A3P5PS and MRS2179, or the allosteric enhancer 2,2'-pyridylisatogen tosylate. A further effect of some antagonists, particularly suramin, was to enhance the depolarization-evoked Ca2+ responses during co-application of an agonist. Of several P2Y1 receptor inhibitors, only SCH202676, which has a proposed allosteric mechanism of action, could block ADP-induced voltage dependent Ca2+ release. CONCLUSIONS AND IMPLICATIONS: The ability of depolarization to potentiate GPCRs at near-threshold agonist concentrations represents a novel mechanism for coincidence detection. Furthermore, the induction and enhancement of voltage-dependent GPCR responses by antagonists has implications for the design of therapeutic compounds. PMID- 18414380 TI - Inhibition of the rapid component of the delayed rectifier potassium current in ventricular myocytes by angiotensin II via the AT1 receptor. AB - BACKGROUND AND PURPOSE: There is increasing evidence that angiotensin II (Ang II) is associated with the occurrence of ventricular arrhythmias. However, little is known about the electrophysiological effects of Ang II on ventricular repolarization. The rapid component of the delayed rectifier K(+) current (I(Kr)) plays a critical role in cardiac repolarization. Hence, the aim of this study was to assess the effect of Ang II on I(Kr) in guinea-pig ventricular myocytes. EXPERIMENTAL APPROACH: The whole-cell patch-clamp technique was used to record I(Kr) in native cardiocytes and in human embryonic kidney (HEK) 293 cells, co transfected with human ether-a-go-go-related gene (hERG) encoding the alpha subunit of I(Kr) and the human Ang II type 1 (AT(1)) receptor gene. KEY RESULTS: Ang II decreased the amplitude of I(Kr) in a concentration-dependent manner with an IC(50) of 8.9 nM. Action potential durations at 50% (APD(50)) and 90% (APD(90)) repolarization were prolonged 20% and 16%, respectively by Ang II (100 nM). Ang II-induced inhibition of the I(Kr) was abolished by the AT(1) receptor blocker, losartan (1 muM). Ang II decreased hERG current in HEK293 cells and significantly delayed channel activation, deactivation and recovery from inactivation. Moreover, PKC inhibitors, stausporine and Bis-1, significantly attenuated Ang II-induced inhibition of I(Kr). CONCLUSIONS AND IMPLICATIONS: Ang II produces an inhibitory effect on I(Kr)/hERG currents via AT(1) receptors linked to the PKC pathway in ventricular myocytes. This is a potential mechanism by which elevated levels of Ang II are involved in the occurrence of arrhythmias in cardiac hypertrophy and failure. PMID- 18414381 TI - Effects of imatinib mesylate on spontaneous electrical and mechanical activity in smooth muscle of the guinea-pig stomach. AB - BACKGROUND AND PURPOSE: Effects of imatinib mesylate, a Kit receptor tyrosine kinase inhibitor, on spontaneous activity of interstitial cells of Cajal (ICC) and smooth muscles in the stomach were investigated. EXPERIMENTAL APPROACH: Effects of imatinib on spontaneous electrical and mechanical activity were investigated by measuring changes in the membrane potential and tension recorded from smooth muscles of the guinea-pig stomach. Its effects on spontaneous changes in intracellular concentration of Ca(2+) ([Ca(2+)](i)) (Ca(2+) transients) were also examined in fura-2-loaded preparations. KEY RESULTS: Imatinib (1-10 microM) suppressed spontaneous contractions and Ca(2+) transients. Simultaneous recordings of electrical and mechanical activity demonstrated that imatinib (1 microM) reduced the amplitude of spontaneous contractions without suppressing corresponding slow waves. In the presence of nifedipine (1 microM), imatinib (10 microM) reduced the duration of slow waves and follower potentials in the antrum and accelerated their generation, but had little affect on their amplitude. In contrast, imatinib reduced the amplitude of antral slow potentials and slow waves in the corpus. CONCLUSIONS AND IMPLICATIONS: Imatinib may suppress spontaneous contractions of gastric smooth muscles by inhibiting pathways that increase [Ca(2+)](i) in smooth muscles rather than by specifically inhibiting the activity of ICC. A high concentration of imatinib (10 microM) reduced the duration of slow waves or follower potentials in the antrum, which reflect activity of ICC distributed in the myenteric layers (ICC-MY), and suppressed antral slow potentials or corporal slow waves, which reflect activity of ICC within the muscle bundles (ICC-IM), presumably by inhibiting intracellular Ca(2+) handling. PMID- 18414382 TI - AM3, a natural glycoconjugate, induces the functional maturation of human dendritic cells. AB - BACKGROUND AND PURPOSE: Dendritic cells (DCs) are dedicated antigen-presenting cells able to initiate specific immune responses and their maturation is critical for the induction of antigen-specific T-lymphocyte responses. Here, we have investigated the effects of Inmunoferon-active principle (AM3), the active agent of a commercial immunomodulatory drug, on human monocyte-derived DCs (MDDCs). EXPERIMENTAL APPROACH: MDDCs derived from healthy and hepatitis C virus (HCV) infected patients were stimulated with AM3. We analysed the expression of cell surface proteins by flow cytometry, that of cytokine production by ELISA, and the expression of chemokines and chemokine receptors by RNase protection assays. T lymphocyte proliferation was assessed in mixed lymphocyte reactions, protein expression by western blot and luciferase-based reporter methods, and Toll-like receptor (TLR)-blocking antibodies were employed to analyse TLR activity. KEY RESULTS: In MDDCs, AM3 induced or enhanced expression of CD54, CD83, CD86, HLA DR, chemokines and chemokine receptors, interleukin (IL)-12p70 and IL-10. Furthermore, AM3 stimulated MDDCs to increase proliferation of allogenic T cells. AM3 triggered nuclear translocation of NF-kappaB and phosphorylation of p38 mitogen-activated protein kinase. AM3 promoted NF-kappaB activation in a TLR-4 dependent manner, and blocking TLR-4 activity attenuated the enhanced expression of CD80, CD83 and CD86 induced by AM3. AM3 enhanced the expression of maturation associated markers in MDDCs from HCV-infected patients and increased the proliferation of T lymphocytes induced by these MDDCs. CONCLUSIONS AND IMPLICATIONS: These results underline the effects of AM3 in promoting maturation of MDDCs and suggest that AM3 might be useful in regulating immune responses in pathophysiological situations requiring DC maturation. PMID- 18414384 TI - Neuropharmacology of addiction--setting the scene. AB - Addiction is a complex disorder, affecting not only the individual addict, but also their family and the community at large. While therapeutic strategies are available for the treatment of some forms of substance abuse/dependence, these are not without problems and are not universally efficacious. Moreover, in some instances (for example, cocaine addiction), there are still no medications specifically registered as treatment options. In this themed issue of the British Journal of Pharmacology, we highlight a number of addictions from a pharmacological perspective, with an emphasis on both mechanism and potential therapeutic approaches that are either under development or reflect preclinical work. As such, the authors endeavour to describe the latest thinking on the neural theory of addiction and corresponding novel pharmacotherapeutic targets, and in this way to set the stage for future advances in research and drug development. In addition, we have also attempted to draw attention to the clinicians' perspective in terms of the interface between basic science and care provision. PMID- 18414383 TI - A neuropeptide-centric view of psychostimulant addiction. AB - Drugs of abuse all share common properties classically observed in human beings and laboratory animals. They enhance neural firing and dopamine tone within the nucleus accumbens and produce progressively greater drug-induced motor responses defined as behavioural sensitization. They produce conditioned place preference, a behavioural model of incentive motivation, which highlights the role of environmental cues in drug addiction. They increase brain reward function as seen by a lowering of intracranial self-stimulation thresholds. And last but not least, they are self-administered, and sometimes even abused, and can trigger reinstatement of drug-seeking behaviour in animals extinguished from drug self administration. It has long been considered that the reinforcing properties of virtually all drugs of abuse, more specifically psychostimulants, are primarily dependent on activation of the mesolimbic dopamine system. However, recent evidence raises the importance of dopamine-independent mechanisms in reward related behaviours. The overwhelming body of evidence that indicates a critical role for the mesolimbic dopamine system in the reinforcing effect of psychostimulants should not mask the key contribution of other modulatory systems in the brain. This review summarizes the complex and subtle role of several neuropeptidergic systems in various aspects of addictive behaviours observed in laboratory animals exposed to psychostimulants. A special emphasis is given to the cannabinoid, opioid, nociceptin/orphanin FQ, corticotropin-releasing factor and hypocretin/orexin systems. The relevance of these systems viewed as potential therapeutic targets for drug addiction is discussed in the light of their narrow pharmacological profile and their effectiveness in preventing drug addiction at doses usually not accompanied by severe side effects. PMID- 18414386 TI - Regression of aortic valve stenosis by ApoA-I mimetic peptide infusions in rabbits. AB - BACKGROUND AND PURPOSE: Aortic valve stenosis (AVS) is the most common valvular heart disease, and standard curative therapy remains open heart surgical valve replacement. The aim of our experimental study was to determine if apolipoprotein A-I (ApoA-I) mimetic peptide infusions could induce regression of AVS. EXPERIMENTAL APPROACH: Fifteen New Zealand White male rabbits received a cholesterol-enriched diet and vitamin D(2) until significant AVS was detected by echocardiography. The enriched diet was then stopped to mimic cholesterol lowering therapy and animals were allocated randomly to receive saline (control group, n=8) or an ApoA-I mimetic peptide (treated group, n=7), three times per week for 2 weeks. Serial echocardiograms and post mortem valve histology were performed. KEY RESULTS: Aortic valve area increased significantly by 25% in the treated group after 14 days of treatment (P=0.012). Likewise, aortic valve thickness decreased by 21% in the treated group, whereas it was unchanged in controls (P=0.0006). Histological analysis revealed that the extent of lesions at the base of valve leaflets and sinuses of Valsalva was smaller in the treated group compared with controls (P=0.032). The treatment also reduced calcification, as revealed by the loss of the positive relationship observed in the control group (r=0.87, P=0.004) between calcification area and aortic valve thickness. CONCLUSIONS AND IMPLICATIONS: Infusions of ApoA-I mimetic peptide lead to regression of experimental AVS. These positive results justify the further testing of high-density lipoprotein (HDL)-based therapies in patients with valvular aortic stenosis. Regression of aortic stenosis, if achieved safely, could transform the clinical treatment of this disease. PMID- 18414387 TI - Diabetic cardiomyopathy: how much does it depend on AGE? AB - Diabetic cardiomyopathy refers to dysfunction of cardiac muscle in patients with diabetes that cannot be directly ascribed to hypertension, coronary heart disease or other defined cardiac abnormalities per se. The development of diabetic cardiomyopathy may involve several distinct mechanisms, including increased formation of advanced glycation end products (AGEs) secondary to hyperglycaemia. AGEs may alter structural proteins and lead to increased arterial and myocardial stiffness. Therefore, therapies that prevent or retard development of AGEs in diabetes may be valuable strategies to treat or prevent diabetic cardiomyopathy. In this issue of British Journal of Pharmacology, Wu and colleagues demonstrate that aminoguanidine (inhibitor of AGE formation and protein cross-linking) treatment of a rat model of type I diabetes (rats made insulin deficient with streptozotocin and nicotinamide treatment) ameliorates detrimental changes in left ventricular structure and function. Results from this study are in agreement with previous investigations, suggesting that aminoguanidine is effective in preventing cardiac hypertrophy and arterial stiffening in experimental animal models of diabetes and emphasize the potential pathogenic role of AGEs in diabetic cardiomyopathy. PMID- 18414385 TI - The endocannabinoid system in brain reward processes. AB - Food, drugs and brain stimulation can serve as strong rewarding stimuli and are all believed to activate common brain circuits that evolved in mammals to favour fitness and survival. For decades, endogenous dopaminergic and opioid systems have been considered the most important systems in mediating brain reward processes. Recent evidence suggests that the endogenous cannabinoid (endocannabinoid) system also has an important role in signalling of rewarding events. First, CB(1) receptors are found in brain areas involved in reward processes, such as the dopaminergic mesolimbic system. Second, activation of CB(1) receptors by plant-derived, synthetic or endogenous CB(1) receptor agonists stimulates dopaminergic neurotransmission, produces rewarding effects and increases rewarding effects of abused drugs and food. Third, pharmacological or genetic blockade of CB(1) receptors prevents activation of dopaminergic neurotransmission by several addictive drugs and reduces rewarding effects of food and these drugs. Fourth, brain levels of the endocannabinoids anandamide and 2-arachidonoylglycerol are altered by activation of reward processes. However, the intrinsic activity of the endocannabinoid system does not appear to play a facilitatory role in brain stimulation reward and some evidence suggests it may even oppose it. The influence of the endocannabinoid system on brain reward processes may depend on the degree of activation of the different brain areas involved and might represent a mechanism for fine-tuning dopaminergic activity. Although involvement of the various components of the endocannabinoid system may differ depending on the type of rewarding event investigated, this system appears to play a major role in modulating reward processes. PMID- 18414389 TI - Cellular and molecular mechanisms responsible for the action of testosterone on human skeletal muscle. A basis for illegal performance enhancement. AB - The popularity of testosterone among drug users is due to its powerful effects on muscle strength and mass. Important mechanisms behind the myotrophic effects of testosterone were uncovered both in athletes using steroids for several years and in short-term controlled studies. Both long-term and short-term steroid usage accentuates the degree of fibre hypertrophy in human skeletal muscle by enhancing protein synthesis. A mechanism by which testosterone facilitates the hypertrophy of muscle fibres is the activation of satellite cells and the promotion of myonuclear accretion when existing myonuclei become unable to sustain further enhancement of protein synthesis. Interestingly, long-term steroid usage also enhances the frequency of fibres with centrally located myonuclei, which implies the occurrence of a high regenerative activity. Under the action of testosterone, some daughter cells generated by satellite cell proliferation may escape differentiation and return to quiescence, which help to replenish the satellite cell reserve pool. However, whether long-term steroid usage induces adverse effects of satellite cells remains unknown. Testosterone might also favour the commitment of pluripotent precursor cells into myotubes and inhibit adipogenic differentiation. The effects of testosterone on skeletal muscle are thought to be mediated via androgen receptors expressed in myonuclei and satellite cells. Some evidence also suggests the existence of an androgen-receptor-independent pathway. Clearly, testosterone abuse is associated with an intense recruitment of multiple myogenic pathways. This provides an unfair advantage over non-drug users. The long-term consequences on the regenerative capacity of skeletal muscle are unknown. PMID- 18414388 TI - Central ghrelin gastroprotection involves nitric oxide/prostaglandin cross-talk. AB - BACKGROUND AND PURPOSE: Ghrelin, a gut-brain peptide, is considered a gastroprotective factor in gastric mucosa. We investigated the role of prostaglandins (PG) and the possible interplay between PGs and nitric oxide (NO) in ghrelin gastroprotection against ethanol (EtOH)-induced gastric lesions. EXPERIMENTAL APPROACH: We examined the effects of (1) central ghrelin (4 mug per rat) injection on PGE(2) accumulation in normal or EtOH-lesioned gastric mucosa, (2) pretreatment with indomethacin (10 mg kg(-1), p.o.), a non-selective cyclooxygenase (COX) inhibitor, and with a selective COX-1, SC560 (5 mg kg(-1), p.o.) or COX-2 inhibitor, celecoxib (3.5 mg kg(-1), p.o.) on ghrelin gastroprotection against 50% EtOH (1 mL per rat)-induced gastric lesions, (3) the NO synthase inhibitor, L-NAME (70 mg kg(-1), s.c), on gastric PGE(2) content in ghrelin-treated rats and (4) central ghrelin on the expression of constitutive and inducible NOS and COX mRNA and on the localization of the immunoreactivity for COX-2 in the gastric mucosa exposed to EtOH. KEY RESULTS: Ghrelin increased PGE(2) in normal mucosa, whereas, it reversed the EtOH-induced PGE(2) surge. Ghrelin had no effect on mucosal COX-1 expression but reduced the EtOH-induced increase in COX-2 expression and immunoreactivity. Indomethacin and SC560, but not celecoxib, removed ghrelin gastroprotection. L-NAME prevented the PGE(2) surge induced by ghrelin and, like indomethacin, reduced EtOH-induced PGE(2) increase. Ghrelin enhanced eNOS expression and reduced iNOS mRNA. CONCLUSIONS AND IMPLICATIONS: This study shows that COX-1-derived PGs are mainly involved in ghrelin gastroprotection and that the constitutive-derived NO together with PGE(2) are involved in ghrelin gastroprotective activity. PMID- 18414390 TI - Chronic heart rate reduction by ivabradine prevents endothelial dysfunction in dyslipidaemic mice. AB - BACKGROUND AND PURPOSE: High resting heart rate is a predictor for total and cardiovascular mortality independent of other risk factors in patients with coronary artery disease. We tested the hypothesis that a reduction of resting heart rate with the cardiac pacemaker I(f) current inhibitor ivabradine prevents the endothelial dysfunction associated with dyslipidaemia. EXPERIMENTAL APPROACH: Three-month-old dyslipidaemic (DL) male mice expressing the human ApoB-100 were assigned or not (DL, n=16), to treatment for 3 months with ivabradine (10 mg kg( 1) d(-1), n=17). Wild-type C57Bl/6 mice (WT, n=15) were used as controls. Heart rate was measured at 3, 4.5 and 6 months. Dilatation to acetylcholine (ACh) of isolated cerebral and renal arteries was investigated at 6 months. KEY RESULTS: Heart rate remained stable in anaesthetized WT mice, increased (25%, P<0.05) with age in DL mice but was limited (11%, P<0.05) by ivabradine. At 6 months, left ventricular maximal pressure was similar in all groups. The minimal and end diastolic left ventricular pressures were increased (P<0.05) in DL (10.2+/-1.0 and 18.7+/-1.4 mm Hg) compared to WT (-0.4+/-0.7 and 6.3+/-1.0 mm Hg) and reduced (P<0.05) by ivabradine (4.2+/-1.3 and 11.5+/-1.5 mm Hg). ACh-induced maximal dilatation was impaired (P<0.05) in renal and cerebral arteries isolated from DL compared to WT (56+/-7 versus 83+/-3% in renal arteries; 22+/-2 versus 42+/-2% in cerebral arteries). Ivabradine completely prevented (P<0.05) this dysfunction in renal and cerebral arteries. CONCLUSIONS AND IMPLICATIONS: Selective heart rate reduction with ivabradine limits cardiac dysfunction and prevents the renovascular and cerebrovascular endothelial dysfunction associated with dyslipidaemia. PMID- 18414391 TI - Bone morphogenetic protein signalling is required for the anti-mitogenic effect of the proteasome inhibitor MG-132 on colon cancer cells. AB - BACKGROUND AND PURPOSE: Inhibition of proteasome has been emerging as a promising approach in pathway-directed cancer therapy. Bone morphogenetic protein (BMP) signalling, which is known to be regulated by the ubiquitin-proteasome pathway in osteoblasts, plays a crucial role in the suppression of gastrointestinal carcinogenesis. Here we sought to elucidate the anti-mitogenic effect of a proteasome inhibitor in relation to BMP signalling in colon cancer. EXPERIMENTAL APPROACH: The effects of the proteasome inhibitor MG-132 on proliferation of SW1116 and HT-29 colon cancer cells were determined by [(3)H]-thymidine incorporation and colony-formation assay. The involvement of BMP signalling in the action of MG-132 was elucidated by western blot, real-time PCR, immunofluorescence and RNA interference. KEY RESULTS: MG-132 significantly suppressed the proliferation of colon cancer SW1116 and HT-29 cells. In this regard, MG-132 activated BMP signalling and this was manifested as an increase in Smad1/5/8 phosphorylation and upregulation of p21(Waf1/Cip1) and p27(Kip1) expression. Knockdown of BMP receptor II abolished Smad1/5/8 phosphorylation, the induction of p21(Waf1/Cip1) and p27(Kip1) and inhibition of cell proliferation induced by MG-132. Further analysis revealed that MG-132 upregulated the expression of BMP1 and BMP2, which are secreted members of the BMP superfamily. Moreover, the expression of Smad6, an intracellular inhibitor of BMP signalling, was suppressed by MG-132. CONCLUSIONS AND IMPLICATIONS: These findings suggest that inhibition of proteasome suppresses the proliferation of colon cancer cells via activation of BMP signalling. They also demonstrate a novel aspect of proteasome function in the regulation of colon cancer cell proliferation. PMID- 18414392 TI - Aldosterone and the autocrine modulation of potassium currents and oxidative stress in the diabetic rat heart. AB - BACKGROUND AND PURPOSE: Aldosterone plays a major role in cardiac pathology. This study was designed to investigate the role of cardiac aldosterone in modulating K(+) currents and oxidative stress in the streptozotocin-induced diabetic rat heart. EXPERIMENTAL APPROACH: Transient and sustained K(+) currents were measured in ventricular myocytes by voltage clamp. Plasma and cellular aldosterone were measured by ELISA. Fluorescent dihydroethidium (DHE) was used to assess superoxide ions as markers of oxidative stress. KEY RESULTS: The mineralocorticoid antagonist spironolactone (1 microM, 5-9 h) significantly augmented both K(+) currents in diabetic males, with a concomitant shortening of the action potential but had no effect in myocytes from control males or from diabetic females. Effects of spironolactone were restored in ovariectomized diabetic females and abolished in orchidectomized diabetic males. The aldosterone synthase inhibitor FAD286 (1 microM, 5-9 h) significantly augmented K(+) currents in cells from diabetic males, but not females. Spironolactone and FAD286 significantly reduced oxidative stress in cells from diabetic males. Plasma aldosterone content was elevated in diabetic males (relative to control), but not in females. Cellular aldosterone was also elevated, but not significantly. The elevation in aldosterone was only partly dependent on a concomitant increase in cellular angiotensin II. CONCLUSIONS AND IMPLICATIONS: A gender-related, sex hormone-dependent elevation in plasma and cardiac cell aldosterone contributed to oxidative stress and to attenuation of K(+) currents in diabetic male rats. Aldosterone may thus contribute to diabetes-associated cardiac arrhythmias. Aldosterone elevation was partly related to levels of angiotensin II, but residual, angiotensin II-independent, aldosterone maintains functional relevance. PMID- 18414393 TI - Standard antiepileptic drugs fail to block epileptiform activity in rat organotypic hippocampal slice cultures. AB - BACKGROUND AND PURPOSE: Earlier studies had demonstrated that tonic-clonic seizure-like events (SLEs) resembling electrographic correlates of limbic seizures in animals and humans can be induced in organotypic hippocampal slice cultures (OHSCs). We have explored OHSCs for their suitability to serve as in vitro models of limbic seizures for studying seizure mechanisms and screening new antiepileptic compounds. EXPERIMENTAL APPROACH: OHSCs were cultivated according to the interface method. Neuronal activity and extracellular potassium concentration were recorded under submerged conditions. SLEs were induced by lowering magnesium concentration or by applying the potassium channel blocker 4 aminopyridine. The effects of standard antiepileptic drugs (AEDs), carbamazepine, phenytoin, valproic acid, clonazepam, diazepam and phenobarbital sodium on SLEs were analysed. KEY RESULTS: In more than 93% of OHSCs, AEDs did not prevent the induction of SLEs or stop ongoing seizure activity even when toxic concentrations were applied. This pharmacoresistance was independent of the method of seizure provocation, postnatal age at explantation (P2-P10) and cultivation time in vitro (2 months). SLEs were reversibly blocked by glutamate antagonists or the GABA(A) agonist muscimol. CONCLUSIONS AND IMPLICATIONS: We present a simple to establish in vitro model of tonic-clonic SLEs that is a priori pharmacoresistant and thus has an advantage over animal models of pharmacoresistant seizures in which responders and non-responders can be sorted out only after an experiment. OHSCs could be suitable for exploring mechanisms of pharmacoresistant seizures and be used for the identification of new anticonvulsive compounds eventually effective in drug refractory epilepsy. PMID- 18414394 TI - The existence of a local 5-hydroxytryptaminergic system in peripheral arteries. AB - BACKGROUND AND PURPOSE: 5-HT is a vasoconstrictor exhibiting enhanced effects in systemic arteries from subjects with cardiovascular disease. The effect of endogenous 5-HT on arteries is controversial, because the concentration of free circulating 5-HT is low and a 5-hydroxytryptaminergic system has not been identified in peripheral arteries. We hypothesized that a local 5 hydroxytryptaminergic system (including 5-HT synthesis, metabolism, uptake and release) with physiological function exists in peripheral arteries. EXPERIMENTAL APPROACH: The presence of key components of a 5-hydroxytryptaminergic system in rat aorta and superior mesenteric artery was examined using western blot analyses, immunohistochemistry and immunocytochemistry. The function of the rate limiting enzyme in 5-HT biosynthesis, tryptophan hydroxylase (TPH), and 5-HT transporter was tested by measuring enzyme activity and 5-HT uptake, respectively. Isometric contraction of arterial strips was used to demonstrate the function of released endogenous 5-HT in arterial tissues. KEY RESULTS: mRNA for TPH-1 was present in arteries, with low levels of TPH protein and TPH activity. Expression and function of MAO A (5-HT metabolizing enzyme) was supported by immunohistochemistry, western analyses and the elevation of concentrations of 5-hydroxyindoleacetic acid (5-HT metabolite) after exposure to exogenous 5-HT. The 5-HT transporter was localized to the plasma membrane of freshly isolated aortic smooth muscle cells. Peripheral arteries actively took up 5-HT in a time-dependent and 5-HT transporter-dependent manner. The 5-HT transporter substrate, (+)-fenfluramine, released endogenous 5-HT from peripheral arteries, which potentiated noradrenaline-induced arterial contraction. CONCLUSIONS AND IMPLICATIONS: This study revealed the existence of a local 5 hydroxytryptaminergic system in peripheral arteries. PMID- 18414395 TI - Oxidative stress and COX cause hyper-responsiveness in vascular smooth muscle of the femoral artery from diabetic rats. AB - BACKGROUND AND PURPOSE: To investigate the dysfunction of vascular smooth muscle in streptozotocin-induced diabetic rats. EXPERIMENTAL APPROACH: Rings without endothelium of femoral arteries were suspended in organ chambers for isometric tension recording. The production of oxygen-derived free radicals was measured with 2',7'-dichlorodihydrofluorescein diacetate using confocal microscopy. The protein expressions were measured by western blotting. KEY RESULTS: The concentration-response curves to U46619 and phenylephrine, but not that to KCl, were shifted to the left, suggesting a hypersensitivity of cell membrane receptors in diabetes. Exogenous oxygen-derived free radicals induced greater vasoconstrictions in the femoral artery from diabetic rats. Chronic treatment with apocynin (inhibitor of NADPH oxidase) and acute exposure to MnTMPyP (SOD/catalase mimetic) normalized the response. The catalase activity and the total glutathione level were reduced in arteries from streptozotocin-treated rats, confirming a redox abnormality. The basal oxidative state was higher in arteries from streptozotocin-treated rats and reduced in arteries from apocynin- and streptozotocin-treated rats, suggesting that the functional changes in diabetes are due to a chronic increase in oxidative stress. In the arteries of streptozotocin-treated rats, inhibitors of COX-1 and/or COX-2 prevented the hypersensitivity and reduced the increase in oxidative stress caused by phenylephrine and U46619, suggesting that both isoforms contribute to the smooth muscle dysfunction. The expression of proteins for COX-1 and COX-2 was increased in arteries of streptozotocin-treated rats and reduced in preparations of apocynin- and streptozotocin-treated rats. CONCLUSIONS AND IMPLICATIONS: Chronic diabetes and the resulting increased oxidative stress activate the production of COX-derived vasoconstrictor prostanoids causing hypersensitivity of vascular smooth muscle. PMID- 18414396 TI - The expression and function of Ca(2+)-sensing receptors in rat mesenteric artery; comparative studies using a model of type II diabetes. AB - BACKGROUND AND PURPOSE: The extracellular calcium-sensing receptor (CaR) in vascular endothelial cells activates endothelial intermediate-conductance, calcium-sensitive K(+) channels (IK(Ca)) indirectly leading to myocyte hyperpolarization. We determined whether CaR expression and function was modified in a rat model of type II diabetes. EXPERIMENTAL APPROACH: Pressure myography, western blotting, sharp microelectrode and K(+)-selective electrode recordings were used to investigate the functional expression of the CaR and IK(Ca) in rat mesenteric arteries. KEY RESULTS: Myocyte hyperpolarization to the CaR activator calindol was inhibited by Calhex 231. U46619-induced vessel contraction elevated the extracellular [K(+)] around the myocytes, and inhibition of this 'K(+) cloud' by iberiotoxin was needed to reveal calindol-induced vasodilatations. These were antagonized by Calhex 231 and significantly smaller in Zucker diabetic fatty rat (ZDF) vessels than in Zucker lean (ZL) controls. Myocyte hyperpolarizations to calindol were also smaller in ZDF than in ZL arteries. In ZDF vessels, endothelial cell CaR protein expression was reduced; IK(Ca) expression was also diminished, but IK(Ca)-generated hyperpolarizations mediated by 1-EBIO were unaffected. CONCLUSIONS AND IMPLICATIONS: The reduced CaR-mediated hyperpolarizing and vasodilator responses in ZDF arteries result from a decrease in CaR expression, rather than from a modification of IK(Ca) channels. Detection of CaR-mediated vasodilatation required the presence of iberiotoxin, suggesting a CaR contribution to vascular diameter, that is, inversely related to the degree of vasoconstriction. Compromise of the CaR pathway would favour the long-term development of a higher basal vascular tone and could contribute to the vascular complications associated with type II diabetes. PMID- 18414397 TI - Discovery and biological characterization of a novel series of androgen receptor modulators. AB - BACKGROUND AND PURPOSE: Selective androgen receptor modulators are of great value in the treatment of prostate cancer. The purpose of this study was to provide a preliminary characterization of a new class of non-steroidal androgen receptor modulators discovered in a high-throughput screening campaign. EXPERIMENTAL APPROACH: Competitive receptor binding, luciferase-based reporter methods, cell proliferation and in vivo assays were employed to evaluate an initial set of compounds from chemistry efforts. KEY RESULTS: Forty-nine analogues from the chemistry efforts showed high affinity binding to androgen receptors, agonist and/or antagonist activities in both CV-1 and MDA-MB-453 transfection assays. A proliferation assay in LNCaP cells also exhibited this profile. A representative of these non-steroidal compounds (compound 21) was devoid of activity at other nuclear receptors (oestrogen, progesterone, glucocorticoid and mineralocorticoid receptors) in the CV-1 co-transfection assay. At the same time, in an immature castrated rat model, it behaved as an androgen receptor antagonist against the growth of prostate, seminal vesicles and levator ani induced by exogenous androgen. Separation of compound 21 into its enantiomers showed that nearly all the androgen receptor modulating activity and binding resided in the dextrorotatory compound (23) while the laevorotatory isomer (22) possessed weak or little effect depending on the cell type studied. CONCLUSIONS AND IMPLICATIONS: These non-steroidal compounds may represent a new class of androgen receptor modulators for the treatment of not only prostate cancer but other clinical conditions where androgens and androgen receptors are involved in the pathological processes. PMID- 18414398 TI - Gonadotropins in doping: pharmacological basis and detection of illicit use. AB - Parenteral administration of human chorionic gonadotropin (hCG) or luteinizing hormone (LH) stimulates the production of testosterone in males and these gonadotropins can therefore be used by athletes to enhance muscle strength. However, they are more expensive and less efficient than testosterone and anabolic steroids. Therefore their main use is probably to stimulate gonadal testosterone production during and after self-administration of testosterone or anabolic steroids. A positive effect of hCG on muscle strength has not been demonstrated in women and elevated concentrations of hCG in females are often caused by pregnancy. The use of gonadotropins is therefore prohibited only in males but not in females. HCG occurs at low but measurable concentrations in plasma and urine of healthy males and can be measured by sensitive methods. However, the characteristics of the method to be used for doping control have not been defined. Virtually all commercially available hCG assays have been designed for determination of hCG in serum rather than urine, which is used for doping control. Methods based on mass spectrometric detection of fragments derived from hCG extracted from urine by immunoadsorption have been developed but their suitability for doping control remains to be determined. The concentrations of LH in serum and urine are variable and more then 10-fold higher than those hCG. It is therefore difficult to detect illicit use of LH. The characteristics and reference values for hCG and LH assays used in doping control and the cutoff values need to be defined. PMID- 18414399 TI - Addiction: the clinical interface. AB - This review gives an overview of what we see as the key issues in the human pharmacology of drugs of addiction. We review evidence of efficacy and mechanisms by which treatments act and point out areas where further work is needed. The role of agonist, partial agonist and antagonist treatments for opioid addiction is detailed and current issues relating to the mechanisms of actions at the receptor level and how to improve on compliance are discussed. The role of the brain dopamine and GABA-A systems in drug dependence is considered in relation to the growing pharmacology of these receptor systems, and the current status of novel preclinical targets reviewed. In addition, the different roles of dynamic and kinetic factors in both addiction and its treatment are discussed in relation to the underlying neuropharmacology of the disorders as defined from human and preclinical studies. Finally, some pointers to future research and especially to drug development by pharma are elaborated. PMID- 18414401 TI - The fibromatosis signature defines a robust stromal response in breast carcinoma. AB - Breast cancer is a heterogeneous disease, and the influence of stromal gene and protein expression patterns on the biological and clinical heterogeneity of the disease is poorly understood. We previously demonstrated that evaluation of the gene expression patterns of two soft-tissue tumors (desmoid-type fibromatosis (DTF) and solitary fibrous tumor) could be used to identify distinct stromal reaction patterns in breast carcinoma. In the current study, we examined four additional data sets obtained from four different institutions and containing gene expression data from a total of 561 breast cancer patients. We identified a core set of 66 DTF-associated genes that were consistently coordinately expressed in a subset of 25-35% of breast cancers. Breast carcinomas defined by high levels of coordinated expression of DTF core genes tend to be lower grade, express estrogen receptor, and show significantly longer survival across the four data sets. Using multiple tissue microarrays of archival breast cancer specimens obtained from a total of 745 patients, we demonstrated that a subset of breast cancers show coordinate expression of DTF core proteins by stromal cells in the tumor microenvironment. We evaluated the protein expression of a single DTF core protein (SPARC) on a tissue microarray with clinical outcome data and demonstrated that breast cancers with strong stromal protein expression of SPARC show a trend for increased survival. Our data demonstrate that the DTF core gene set is a robust descriptor of a distinct stromal response that is associated with improved clinical outcome in breast cancer patients. PMID- 18414400 TI - Cellular neuroadaptations to chronic opioids: tolerance, withdrawal and addiction. AB - A large range of neuroadaptations develop in response to chronic opioid exposure and these are thought to be more or less critical for expression of the major features of opioid addiction: tolerance, withdrawal and processes that may contribute to compulsive use and relapse. This review considers these adaptations at different levels of organization in the nervous system including tolerance at the mu-opioid receptor itself, cellular tolerance and withdrawal in opioid sensitive neurons, systems tolerance and withdrawal in opioid-sensitive nerve networks, as well as synaptic plasticity in opioid sensitive nerve networks. Receptor tolerance appears to involve enhancement of mechanisms of receptor regulation, including desensitization and internalization. Adaptations causing cellular tolerance are more complex but several important processes have been identified including upregulation of cAMP/PKA and cAMP response element-binding signalling and perhaps the mitogen activated PK cascades in opioid sensitive neurons that might not only influence tolerance and withdrawal but also synaptic plasticity during cycles of intoxication and withdrawal. The potential complexity of network, or systems adaptations that interact with opioid-sensitive neurons is great but some candidate neuropeptide systems that interact with mu-opioid sensitive neurons may play a role in tolerance and withdrawal, as might activation of glial signalling. Implication of synaptic forms of learning such as long term potentiation and long term depression in opioid addiction is still in its infancy but this ultimately has the potential to identify specific synapses that contribute to compulsive use and relapse. PMID- 18414402 TI - Out of sequence: how consumer genomics could displace clinical genetics. PMID- 18414404 TI - Segmental patterning of the vertebrate embryonic axis. AB - The body axis of vertebrates is composed of a serial repetition of similar anatomical modules that are called segments or metameres. This particular mode of organization is especially conspicuous at the level of the periodic arrangement of vertebrae in the spine. The segmental pattern is established during embryogenesis when the somites--the embryonic segments of vertebrates--are rhythmically produced from the paraxial mesoderm. This process involves the segmentation clock, which is a travelling oscillator that interacts with a maturation wave called the wavefront to produce the periodic series of somites. Here, we review our current understanding of the segmentation process in vertebrates. PMID- 18414403 TI - Advances in autism genetics: on the threshold of a new neurobiology. AB - Autism is a heterogeneous syndrome defined by impairments in three core domains: social interaction, language and range of interests. Recent work has led to the identification of several autism susceptibility genes and an increased appreciation of the contribution of de novo and inherited copy number variation. Promising strategies are also being applied to identify common genetic risk variants. Systems biology approaches, including array-based expression profiling, are poised to provide additional insights into this group of disorders, in which heterogeneity, both genetic and phenotypic, is emerging as a dominant theme. PMID- 18414405 TI - Ontogeny of sensorimotor gating and immune impairment induced by prenatal immune challenge in rats: implications for the etiopathology of schizophrenia. AB - It has been hypothesized that the maternal immune response to infection may influence fetal brain development and lead to schizophrenia. Animal experimentation has supported this notion by demonstrating altered sensorimotor gating (prepulse inhibition, PPI) in adult rats prenatally exposed to an immune challenge. In the present study, pregnant rats were exposed to the bacterial endotoxin lipopolysaccharide (LPS) throughout gestation and the offspring were examined by evaluating the PPI, dopaminergic function, brain protein expression and cytokine serum levels from weaning to late adulthood. Prenatal LPS exposure induced a deficit in PPI that emerged at 'puberty' and that persisted throughout adult life. This prenatal insult caused age-specific changes in accumbal dopamine levels and in synaptophysin expression in the frontal cortex. Moreover, serum cytokine levels were altered in an age- and cytokine-dependent manner. Here we show that prenatal LPS administration throughout pregnancy causes maturation dependent PPI deficits and age-dependent alterations in dopamine activity, as well as in synaptophysin expression and cytokine levels. PMID- 18414406 TI - Genetic variation in the CHRNA5 gene affects mRNA levels and is associated with risk for alcohol dependence. AB - Alcohol dependence frequently co-occurs with cigarette smoking, another common addictive behavior. Evidence from genetic studies demonstrates that alcohol dependence and smoking cluster in families and have shared genetic vulnerability. Recently a candidate gene study in nicotine dependent cases and nondependent smoking controls reported strong associations between a missense mutation (rs16969968) in exon 5 of the CHRNA5 gene and a variant in the 3'-UTR of the CHRNA3 gene and nicotine dependence. In this study we performed a comprehensive association analysis of the CHRNA5-CHRNA3-CHRNB4 gene cluster in the Collaborative Study on the Genetics of Alcoholism (COGA) families to investigate the role of genetic variants in risk for alcohol dependence. Using the family based association test, we observed that a different group of polymorphisms, spanning CHRNA5-CHRNA3, demonstrate association with alcohol dependence defined by Diagnostic and Statistical Manual of Mental Disorders, 4th edn (DSM-IV) criteria. Using logistic regression we replicated this finding in an independent case-control series from the family study of cocaine dependence. These variants show low linkage disequilibrium with the SNPs previously reported to be associated with nicotine dependence and therefore represent an independent observation. Functional studies in human brain reveal that the variants associated with alcohol dependence are also associated with altered steady-state levels of CHRNA5 mRNA. PMID- 18414407 TI - Antipsychotic drug mechanisms: links between therapeutic effects, metabolic side effects and the insulin signaling pathway. AB - The exact therapeutic mechanism of action of antipsychotic drugs remains unclear. Recent evidence has shown that second-generation antipsychotic drugs (SGAs) are differentially associated with metabolic side effects compared to first generation antipsychotic drugs (FGAs). Their proclivity to cause metabolic disturbances correlates, to some degree, with their comparative efficacy. This is particularly the case for clozapine and olanzapine. In addition, the insulin signaling pathway is vital for normal brain development and function. Abnormalities of this pathway have been found in persons with schizophrenia and antipsychotic drugs may ameliorate some of these alterations. This prompted us to hypothesize that the therapeutic antipsychotic and adverse metabolic effects of antipsychotic drugs might be related to a common pharmacologic mechanism. This article reviews insulin metabolism in the brain and related abnormalities associated with schizophrenia with the goals of gaining insight into antipsychotic drug effects and possibly also into the pathophysiology of schizophrenia. Finally, we speculate about one potential mechanism of action (that is, functional selectivity) that would be consistent with the data reviewed herein and make suggestions for the future investigation that is required before a therapeutic agent based on these data can be realized. PMID- 18414408 TI - The serotonin transporter genotype is associated with intermediate brain phenotypes that depend on the context of eliciting stressor. AB - A variant allele in the promoter region of the serotonin transporter gene, SLC6A4, the s allele, is associated with increased vulnerability to develop anxiety-related traits and depression. Furthermore, functional magnetic resonance imaging (fMRI) studies reveal that s carriers have increased amygdala reactivity in response to aversive stimuli, which is thought to be an intermediate phenotype mediating the influences of the s allele on emotionality. We used high-resolution microPET [18F]fluoro-2-deoxy-D-glucose (FDG) scanning to assess regional brain metabolic activity in rhesus monkeys to further explore s allele-related intermediate phenotypes. Rhesus monkeys provide an excellent model to understand mechanisms underlying human anxiety, and FDG microPET allows for the assessment of brain activity associated with naturalistic environments outside the scanner. During FDG uptake, monkeys were exposed to different ethologically relevant stressful situations (relocation and threat) as well as to the less stressful familiar environment of their home cage. The s carriers displayed increased orbitofrontal cortex activity in response to both relocation and threat. However, during relocation they displayed increased amygdala reactivity and in response to threat they displayed increased reactivity of the bed nucleus of the stria terminalis. No increase in the activity of any of these regions occurred when the animals were administered FDG in their home cages. These findings demonstrate context-dependent intermediate phenotypes in s carriers that provide a framework for understanding the mechanisms underlying the vulnerabilities of s-allele carriers exposed to different types of stressors. PMID- 18414409 TI - Low expression of gamma-glutamyl hydrolase mRNA in primary colorectal cancer with the CpG island methylator phenotype. AB - The CpG island methylator phenotype (CIMP+) in colorectal cancer (CRC) is defined as concomitant and frequent hypermethylation of CpG islands within gene promoter regions. We previously demonstrated that CIMP+ was associated with elevated concentrations of folate intermediates in tumour tissues. In the present study, we investigated whether CIMP+ was associated with a specific mRNA expression pattern for folate- and nucleotide-metabolising enzymes. An exploratory study was conducted on 114 CRC samples from Australia. mRNA levels for 17 genes involved in folate and nucleotide metabolism were measured by real-time RT-PCR. CIMP+ was determined by real-time methylation-specific PCR and compared to mRNA expression. Candidate genes showing association with CIMP+ were further investigated in a replication cohort of 150 CRC samples from Japan. In the exploratory study, low expression of gamma-glutamyl hydrolase (GGH) was strongly associated with CIMP+ and CIMP+-related clinicopathological and molecular features. Trends for inverse association between GGH expression and the concentration of folate intermediates were also observed. Analysis of the replication cohort confirmed that GGH expression was significantly lower in CIMP+ CRC. Promoter hypermethylation of GGH was observed in only 5.6% (1 out of 18) CIMP+ tumours and could not account for the low expression level of this gene. CIMP+ CRC is associated with low expression of GGH, suggesting involvement of the folate pathway in the development and/or progression of this phenotype. Further studies of folate metabolism in CIMP+ CRC may help to elucidate the aetiology of these tumours and to predict their response to anti-folates and 5-fluorouracil/leucovorin. PMID- 18414411 TI - Ribonucleotide reductase subunits M1 and M2 mRNA expression levels and clinical outcome of lung adenocarcinoma patients treated with docetaxel/gemcitabine. AB - Ribonucleotide reductase subunits M1 (RRM1) and M2 (RRM2) are involved in the metabolism of gemcitabine (2',2'-difluorodeoxycytidine), which is used for the treatment of nonsmall cell lung cancer. The mRNA expression of RRM1 and RRM2 in tumours from lung adenocarcinoma patients treated with docetaxel/gemcitabine was assessed and the results correlated with clinical outcome. RMM1 and RMM2 mRNA levels were determined by quantitative real-time PCR in primary tumours of previously untreated patients with advanced lung adenocarcinoma who were subsequently treated with docetaxel/gemcitabine. Amplification was successful in 42 (79%) of 53 enrolled patients. Low levels of RRM2 mRNA were associated with response to treatment (P< 0.001). Patients with the lowest expression levels of RRM1 had a significantly longer time to progression (P=0.044) and overall survival (P=0.02) than patients with the highest levels. Patients with low levels of both RRM1 and RRM2 had a significantly higher response rate (60 vs 14.2%; P=0.049), time to progression (9.9 vs 2.3 months; P=0.003) and overall survival (15.4 vs 3.6; P=0.031) than patients with high levels of both RRM1 and RRM2. Ribonucleotide reductase subunit M1 and RRM2 mRNA expression in lung adenocarcinoma tumours is associated with clinical outcome to docetaxel/gemcitabine. Prospective studies are warranted to evaluate the role of these markers in tailoring chemotherapy. PMID- 18414410 TI - Chromosomal radiosensitivity in head and neck cancer patients: evidence for genetic predisposition? AB - The association between chromosomal radiosensitivity and genetic predisposition to head and neck cancer was investigated in this study. In all, 101 head and neck cancer patients and 75 healthy control individuals were included in the study. The G(2) assay was used to measure chromosomal radiosensitivity. The results demonstrated that head and neck cancer patients had a statistically higher number of radiation-induced chromatid breaks than controls, with mean values of 1.23 and 1.10 breaks per cell, respectively (P<0.001). Using the 90th percentile of the G(2) scores of the healthy individuals as a cutoff value for chromosomal radiosensitivity, 26% of the cancer patients were radiosensitive compared with 9% of the healthy controls (P=0.008). The mean number of radiation-induced chromatid breaks and the proportion of radiosensitive individuals were highest for oral cavity cancer patients (1.26 breaks per cell, 38%) and pharynx cancer patients (1.27 breaks per cell, 35%). The difference between patients and controls was most pronounced in the lower age group (or=8 h: OR, 2.93 (95% CI: 1.39, 6.16)) were more likely to be obese, compared to participants who slept 7 to <8 h. BMI increased with 0.59 kg m(-2) per standard deviation of sleep fragmentation (95% CI: 0.34, 0.84). After adjustment for sleep fragmentation, the association between short sleep and obesity was no longer significant. Exclusion of participants with probable sleep apnea only marginally changed these associations. Self-reported habitual sleep duration was not associated with BMI or obesity. CONCLUSIONS: Sleep duration, as measured with actigraphy, had a U-shaped relationship with BMI and obesity in an elderly population. A highly fragmented sleep is associated with a higher BMI and a higher risk of obesity, and may explain why short sleep is related to obesity. To preclude bias that can be introduced by self-report measures of sleep duration, using multiple measures of sleep parameters is recommended in future research. PMID- 18414420 TI - Obesity effects on depression: systematic review of epidemiological studies. AB - BACKGROUND: Obesity is a well-known cause of cardiovascular disease burden and premature death, but effects on psychological morbidity remain uncertain. This article reports findings following a systematic review of epidemiological studies to determine whether obesity causes depression. METHODS: Multiple databases were searched for English-language studies of etiology of obesity (exposure variable, analyzed as an ordered category) on depression outcomes (dependent variables, continuous or categorical). Studies in children and in women during pregnancy or postpartum were excluded, as were nonrepresentative cross-sectional studies. Searches and identification of studies for inclusion were performed by EA, whereas a descriptive synthesis of important study characteristics was undertaken independently by us. RESULTS: We reviewed 24 out of approximately 4500 potentially relevant studies; 4 were prospective cohort studies and 20 were cross sectional studies (10 from the United States). Effect measures reported in all prospective cohort studies were consistent and suggested that obesity may increase the odds of future depression outcomes (symptoms or nonclinical diagnosis of depression). Effect measures reported in most cross-sectional studies from the United States supported the hypothesized association between obesity and prevalence of depression outcomes for women but not men, in contrast most cross-sectional studies from populations other than the United States consistently failed to find such associations. CONCLUSIONS: Overall, there is a weak level of evidence supporting the hypothesis that obesity increases the incidence of depression outcomes. Few high-quality prospective cohort studies exist, and cross-sectional studies account for the vast body of published evidence, and therefore firm conclusions for causality cannot yet be drawn. Our finding warrants additional high-quality etiological research on this topic. PMID- 18414419 TI - Candidate genes for obesity revealed from a C57BL/6J x 129S1/SvImJ intercross. AB - OBJECTIVE: To identify the genes controlling body fat, we carried out a quantitative trait locus (QTL) analysis using C57BL/6J (B6) and 129S1/SvImJ (129) mice, which differ in obesity susceptibility after consuming an atherogenic diet. METHODS: Mice were fed chow until 8 weeks and an atherogenic diet from 8 to 16 weeks; body fatness was measured by X-ray absorptiometry in 528 (B6 x 129) F(2) at 8 and 16 weeks. A high-density genome scan was performed using 508 polymorphic markers. After identifying the genetic loci, we narrowed the QTL using comparative genomics and bioinformatics. RESULTS: The percentage of body fat was significantly linked to loci on chromosomes (Chr) 1 (22, 68 and 173 Mb), 4 (74 Mb), 5 (73 Mb), 7 (88 Mb), 8 (43 and 80 Mb), 9 (55 Mb), 11 (115 Mb) and 12 (32 Mb); three suggestive loci on Chrs 6 (76 Mb), 9 (30 Mb) and 16 (26 Mb) and two pairs of interacting loci (Chr 2 at 99.8 Mb with Chr 7; Chr 1 at 68 Mb with Chr 11). Comparative genomics narrowed the QTL intervals by 20-57% depending on the chromosome; in most cases, haplotype analysis further narrowed them by about 90%. CONCLUSIONS: Our analysis identified 15 QTL for percentage of body fat. We narrowed the QTL using comparative genomics and haplotype analysis and suggest several candidate genes: Apcs on Chr 1, Ppargc1a on Chr 5, Ucp1 on Chr 8, Angptl6 on Chr 9 and Lpin1 on Chr 12. PMID- 18414421 TI - Weighing obesity stigma: the relative strength of different forms of bias. AB - OBJECTIVE: To compare the strength of weight bias to other common biases, and to develop a psychometrically sound measure to assess and compare bias against different targets. SUBJECTS: A total of 368 university students (75.4% women, 47.6% white, mean age: 21.53 years, mean body mass index (BMI): 23.01 kg/m(2)). MEASUREMENTS: A measure was developed to assess bias against different targets. Three versions of the universal measure of bias (UMB) were developed and validated, each focusing on either 'fat,' 'gay' or 'Muslim' individuals. These were administered to participants, along with two established scales of bias against each target and a measure of socially desirable response style. RESULTS: The UMB demonstrated good internal consistency, appropriate item-total and inter item correlations, and a clear factor structure suggesting components of Negative Judgment, Distance, Attraction and Equal Rights. Construct validity was indicated by strong correlations between established measures of bias and each corresponding version of the new scale. In contrast to previously established measures of weight bias, the new measure was independent of socially desirable response style. Although homosexual orientation was associated with lower gay bias (P<0.05), greater BMI was not associated with any decrease in weight bias. When comparing the relative strength of bias against different targets, weight bias was significantly greater than bias against both gays and Muslims (P<0.001). CONCLUSION: Weight bias is significantly stronger than other major targets of bias. This is the first study to develop a universal measure to assess bias against different targets. The excellent psychometric properties of this measure will permit further investigation into the relative severity of different types of prejudice over time and across samples. The present findings suggest that the pervasive discrimination against obese individuals may be more socially acceptable than discrimination against other groups. PMID- 18414422 TI - Foot morphology of normal, underweight and overweight children. AB - OBJECTIVE: Due to the fact that there is a global increase in obesity, knowledge about the impact of obesity on the development of a child's foot is of great importance for orthopaedic and paediatric physicians with regard to prevention, clinical treatment and management. Therefore, the purpose of this study was to investigate the influence of body mass on the development of a child's foot based on a foot type classification. METHODS: The feet of 1450 boys and 1437 girls aged 2-14 years were measured using a three-dimensional (3D) foot scanner (Pedus, Human Solutions Inc., Germany) in a bipedal upright position. Twelve relevant 3D foot measures were recorded, as well as the children's age, gender, height and mass. Factor analysis of principal components was used to obtain a smaller number of independent and standardized variables. The variables were used for cluster analysis to classify the children's feet. RESULTS: Five foot types were identified: flat, robust, slender, short and long feet. There were significant differences among foot types with respect to the children's body mass index. Normal weight children displayed an almost equal distribution of all foot types throughout childhood. Flat and robust feet were more common in overweight children, whereas underweight children showed more slender and long feet. CONCLUSION: The influence of excess, as well as deficient mass could be verified for the comprehensive foot morphology based on a foot type classification. Subsequently, foot discomfort as a result of various musculoskeletal disorders may develop. In turn, this might keep the children from being active and therefore reinforce the risk of developing obesity. However, there is still a lack of information regarding these relationships, which needs to be determined. This knowledge may help prevent orthopaedic foot problems and injuries. PMID- 18414423 TI - Waist-to-height ratio: a simple option for determining excess central adiposity in young people. AB - Waist circumference is recommended as a means of identifying people at risk of morbidity associated with central adiposity. Yet, there are no universally agreed cut-points to determine when a waist circumference is too large in young people. In this study we examined the relation between sex- and age-specific waist circumference cut-points, the waist-to-height ratio (WHtR) cut-point of <0.5 and cardiovascular disease (CVD) risk clustering in 164 young people, mean age 14.9+/ 0.2 years (mean+/-s.d.). In total 19 (11.6%) of the sample were identified as having CVD risk clustering. These young people were significantly (P<0.001) heavier and had higher body mass index (BMI) and waist circumference z-scores compared to those without CVD risk clustering. The WHtR cut-point of 0.5 estimated CVD risk clustering to a similar extent to sex- and age-adjusted cut points for waist circumference and BMI. Young people with excess central adiposity (WHtR> or =0.5) were 11 times (OR 11.4, P<0.001), more likely to have CVD risk clustering compared to those who did not have excess central adiposity. The WHtR has several advantages; it is easy to calculate, does not require sex- and age-specific centiles and as has been previously suggested, it is a simple message, easily understood by clinicians and families, to 'keep your waist circumference to less than half your height'. PMID- 18414424 TI - Decorin is a secreted protein associated with obesity and type 2 diabetes. AB - OBJECTIVE: To characterize the expression of the small leucine-rich glycoprotein decorin in adipose tissue. DESIGN: Real-time PCR was used to measure decorin gene expression in adipose tissue from normal glucose tolerant (NGT), impaired glucose tolerant and type 2 diabetic (T2D) Psammomys obesus. Adipose tissue was fractionated to determine which cells were responsible for decorin expression. The location of decorin protein expression in adipose tissue was determined using immunohistochemistry. Real-time PCR was used to measure decorin mRNA levels in human adipose tissue from 16 insulin-sensitive, 16 insulin-resistant and 6 T2D human subjects. Circulating plasma decorin concentrations were measured by enzyme linked immunosorbent assay in 145 NGT and 141 T2D human individuals from a large scale epidemiological study in Mauritius. RESULTS: Decorin mRNA was found to be highly expressed in adipose tissue, and decorin gene expression was significantly higher in visceral than that in subcutaneous adipose tissue depots in both P. obesus and human subjects (P=0.002 and P=0.001, respectively). Decorin mRNA was predominantly expressed by stromal/vascular cells of adipose tissue, and decorin protein in adipose tissue was primarily detected adjacent to blood vessels. Circulating plasma decorin levels in humans were elevated by 12% in T2D (P=0.049) compared to NGT subjects. There was a significant independent correlation between plasma decorin levels and waist-to-hip ratio (WHR, P=0.024). In male subjects, plasma decorin levels were significantly correlated with WHR (P=0.006), and fasting and 2-h glucose levels in an oral glucose tolerance test (P=0.027 and P=0.001, respectively). CONCLUSIONS: Decorin expression in adipose tissue was markedly upregulated in the obese state and may therefore play a role in adipose tissue homeostasis or in pathophysiology associated with obesity. PMID- 18414425 TI - The effects of functional electrical stimulation leg cycle ergometry training on arterial compliance in individuals with spinal cord injury. AB - STUDY DESIGN: A prospective intervention of functional electrical stimulation leg cycle ergometry (FES-LCE) of four women with spinal cord injury (SCI). OBJECTIVE: To evaluate the effect of FES-LCE training on arterial compliance in individuals with chronic SCI of traumatic origin. SETTING: Tertiary rehabilitation center in Canada. METHODS: Large and small artery compliance were measured at the radial artery before and after a 3-month training program using FES-LCE. RESULTS: There was no significant change in large artery compliance after FES-LCE (16.0+/-4.2 to 16.8+/-6.1 ml mm Hg(-1) x 10, P=NS). There was a marked (63%) increase in small artery compliance after the FES training program (4.2+/-1.8 to 6.9+/-3.2 ml mm Hg(-1) x 100, P<0.05). CONCLUSION: It appears that FES-LCE is effective in improving small artery compliance in females with SCI. PMID- 18414426 TI - C-Reactive protein in adults with chronic spinal cord injury: increased chronic inflammation in tetraplegia vs paraplegia. AB - STUDY DESIGN: Cross-sectional. OBJECTIVES: In community-dwelling adults with chronic spinal cord injury (SCI), to (1) quantify C-reactive protein (CRP), a marker of inflammation and cardiovascular disease (CVD) risk; (2) determine factors associated with CRP. SETTING: Hamilton, Ontario, Canada. METHODS: We examined CVD risk factors in 69 participants. Measurements included length, weight, waist circumference, blood pressure, percent fat mass (bioelectrical impedance analysis) and fasting blood parameters (high-sensitivity CRP, lipids, insulin, glucose, insulin resistance by homeostasis model assessment (HOMA)). RESULTS: Mean CRP of the group was 3.37+/-2.86 mg-l(-1), consistent with the American Heart Association (AHA) definition of high risk of CVD. CRP was 74% higher in persons with tetraplegia (4.31+/-2.97) than those with paraplegia (2.47+/-2.47 mg l(-1), P=0.002), consistent with high CVD risk. Participants with high CRP (3.1-9.9 mg l(-1)) had greater waist circumference, BMI, percent fat mass and HOMA values than those with lower CRP (< or =3.0 mg l(-1), all P<0.05). LogCRP was independently correlated with waist circumference (r=0.612), logTriglycerides (r=0.342), logInsulin (r=0.309) and logHOMA (r=0.316, all P<0.05). Only level of lesion and waist circumference remained significantly associated with logCRP when variables with significant bivariate correlations were included in multiple regression analysis. CONCLUSION: Mean CRP values in this sample of adults with chronic SCI were consistent with the AHA classification of high CVD risk, especially those of persons with tetraplegia. Level of lesion and waist circumference are independently associated with CRP in this population. PMID- 18414427 TI - The effect of extracorporeal shock wave lithotripsy on the rat spinal cord. AB - STUDY DESIGN: Experimental study. OBJECTIVES: To determine the effects of extracorporeal shock wave lithotripsy (ESWL) on the rat spinal cord. METHODS: Animals were randomly divided into three groups. Groups 1 and 2 consisted of five rats each that underwent ESWL (2000 impulses at 15 kV and 2000 impulses at 18 kV, respectively) and group 3 contained five control rats (no shock wave treatment). ESWL-treated and control rats were compared with regard to light and electron microscopic findings of the adjacent spinal cord. RESULTS: Gross neurological outcomes were normal in all groups. Light microscopic examination of group 1 showed extensive extravasation of red blood cells over all the interstitial spaces. Group 2 also had haemorrhagic areas and an irregular organization of axons in the white matter. Transmission electron microscopic examination of group 1 indicated extravasated red blood cells through the endothelium and swollen axoplasm, degenerated mitochondria, destruction of myelin sheaths and a slight increase in the number of lysosomes. Extravasated red blood cells were also seen in group 2. The axoplasmic mitochondria were enlarged, but no sign of mitochondrial degeneration was observed. Lamellar degeneration of myelin sheaths and abundant lysosomes were more predominant in group 2 than in group 1. CONCLUSION: Extracorporeal shock wave lithotripsy caused not only haemorrhage but also damage to neuronal structures except the nucleus. Our findings showed that higher-energy ESWL caused more myelin degeneration in the spinal cord. PMID- 18414428 TI - Effect of intrathecal papaverine on blood flow and secondary injury in injured cord. AB - STUDY DESIGN: Experimental laboratory investigations with a model of neurotrauma in Macaca rhesus. OBJECT: The present study evaluates whether intrathecal papaverine induces changes in spinal cord blood flow (SCBF) of injured spinal cord and prevents secondary injury. SETTING: Institute of Spinal Cord Injury, Sun Yat-sen University, China. METHODS: After laminectomy was performed and contusive spinal cord injuries were induced in adult female Macaca rhesus, three received intrathecal papaverine, and three received saline 0.9% for control. SCBF was registered by laser-Doppler recording technique continuously for 180 min after injection. Histological analyses and microvessel density (MVD) were used for evaluation of spinal cord injury, and the percentage of spared spinal cord area was calculated. RESULTS: Mean arterial blood pressure showed no significant change in both groups. In the papaverine group, SCBF recovered to 81.35+/-7.8% of baseline at 15 min, 75.24+/-6.3% at 30 min, 73.38+/-2.3% at 90 min and 72.57+/ 4.1% at 180 min after the completion of infusion. SCBF was significantly higher than the control groups (P<0.01). There was no occlusion of the arteries, but occluded veins were identified at the injured site. The MVD in the spinal cord of the control group was significantly lesser than the papaverine group (P<0.01). Luxol Fast Blue staining showed that intrathecal papaverine reduced myelin loss in the lesion 2 weeks after injury (P<0.05). CONCLUSION: Intrathecal administration of papaverine increased SCBF in non-human primates. It is likely that the effects of papaverine can reduce secondary injury in spinal cord injured Macaca rhesus. PMID- 18414429 TI - Prolactin can modulate CD4+ T-cell response through receptor-mediated alterations in the expression of T-bet. AB - Low-dose prolactin induces proinflammatory responses and antibody production, whereas high-dose prolactin suppresses these responses. Mechanisms for these opposing effects remain incompletely defined. We have previously demonstrated that T-bet, a key transcription factor directing T helper type 1 inflammatory responses, is regulated by female steroid hormones in human mucosal epithelial cells via Stat1 and 5 pathways. T-bet was also modulated in a CD4+ T cell line by prolactin exposure. Prolactin rapidly induced T-bet transcription through phosphorylation of JAK2 and Stat5, but not Stat1. Phosphorylated Stat5 then bound to the T-bet regulatory region. These effects were weaker with high-dose prolactin exposures. Upon long-term prolactin exposure, low-dose prolactin induced T-bet expression, whereas high-dose prolactin tended to suppress it. Prolactin induced the suppressors of cytokine signaling (SOCS) 1 and 3 in a dose dependent manner. With high-dose exposure, this was associated with an inhibition of the phosphorylation of T-bet regulatory region-bound Stat5. Further, the dose dependent prolactin effects on T-bet expression were confirmed in murine primary CD4+ T cells. These data suggest that the divergent immune effects of low- and high-dose prolactin may involve modulation of T-bet and alterations in the balance of the prolactin/JAK2/Stat5 and the prolactin/SOCS1 and 3 pathways. PMID- 18414430 TI - Demystifying the development of dendritic cell subtypes, a little. AB - The broadest definition of dendritic cells (DCs) is white blood cells that can take up antigen, process it and then present antigen-derived peptides to activate cognate naive T cells. Although this definition is by no means perfect, it is nevertheless now textbook. The source of frustration more recently has focused on other issues, including the distinction of the DC subtypes, their differential roles in the immune system, their lineage relationship to each other (and other leukocytes) and whether the mouse and human DC findings overlap. Here, I condense the classification of DCs in both the steady state versus infection, with primary focus in the mouse. Emphasis is then given to debates surrounding the in vivo pathways of DC differentiation in different conditions, which culture models best represent these processes (fms-like tyrosine kinase 3 ligand versus granulocyte macrophage colony-stimulating factor), and what the human and mouse DC subtype equivalents might be. In addition, a model termed 'graded' commitment is proposed that, as a departure from the classic binary models of hematopoiesis, attempts to explain the recent clonal data where subtype-specific DC precursors branch from this pathway. PMID- 18414431 TI - Probing in vivo dendritic cell functions by conditional cell ablation. AB - Dendritic cells (DCs) play a central role in T-cell activation and the control of the inherent autoreactivity of the T-cell compartment. Pleiotropic DC functions are likely associated with discrete DC subsets. However, the latter remain largely defined by phenotype and unique anatomic location, rather than function. The investigation of DC involvement in complex phenomena that rely on multicellular interactions, such as immuno-stimulation and tolerization calls for an assessment of DC functions within physiological context. Given the highly dynamic DC compartment, the method of choice to study in vivo DC functions is their conditional ablation in the intact organism. Here, we summarize the recent progress in this field highlighting pitfalls and prospects of the approach. PMID- 18414433 TI - Nephrologists Sans Frontieres: a renal clinician meets comparative physiology on the coast of Maine. PMID- 18414434 TI - Fluid loss from the peritoneal cavity by back-filtration through the small pores of the three-pore model. AB - The partitioning of fluid flows among small and ultrasmall pores of the three pore model in peritoneal dialysis has been traditionally assessed using 4-hour dwells with 3.86% glucose solutions. Under these conditions, however, back filtration through small pores has been hard to demonstrate. As nicely shown by Asghar and Davies, however, the use of low-concentration (1.36%) glucose-based solutions allows accurate studies of the partitioning of fluid flows from the peritoneal cavity under conditions of fluid loss. PMID- 18414435 TI - Quantifying collagen in mouse kidneys. AB - Identification of genes underlying quantitative trait loci (QTLs) represents a formidable task. Recent advances in the annotation of the mouse genome and the availability of newly developed mouse strains should boost gene discovery. Kato and colleagues report the localization of a QTL for collagen type I deposition in kidneys on mouse chromosome 2, representing the first step for the identification of a gene involved in interstitial fibrosis, an important pathway in progression to end-stage renal failure in many kidney disorders. PMID- 18414436 TI - Role for alkaline phosphatase as an inducer of vascular calcification in renal failure? AB - Vascular calcification is associated with increased cardiovascular morbidity and mortality. A number of calcification inhibitors have been defined recently, including inorganic pyrophosphate (PP(i)), an important physicochemical inhibitor of hydroxyapatite crystal growth. Increased hydrolysis of PP(i) by tissue nonspecific alkaline phosphatase (TNAP) may occur in renal failure and act to enhance mineralization of vessels. PMID- 18414439 TI - Better blood pressure control is not causally linked to intradialytic hypotension. PMID- 18414441 TI - Lack of mortality benefit with sevelamer. PMID- 18414442 TI - Interstitial acidosis around the collecting ducts in dRTA. PMID- 18414444 TI - Antiproteinuric effects of cilnidipine. PMID- 18414445 TI - Renal failure with intracapillary thrombi. Lipoprotein glomerulopathy. PMID- 18414446 TI - Hydronephrosis caused by fecal impaction. PMID- 18414447 TI - Enhanced efficiency of generating induced pluripotent stem (iPS) cells from human somatic cells by a combination of six transcription factors. PMID- 18414448 TI - Prostate cancer and metastasis initiating stem cells. AB - Androgen refractory prostate cancer metastasis is a major clinical challenge. Mechanism-based approaches to treating prostate cancer metastasis require an understanding of the developmental origin of the metastasis-initiating cell. Properties of prostate cancer metastases such as plasticity with respect to differentiated phenotype and androgen independence are consistent with the transformation of a prostate epithelial progenitor or stem cell leading to metastasis. This review focuses upon current evidence and concepts addressing the identification and properties of normal prostate stem or progenitor cells and their transformed counterparts. PMID- 18414449 TI - Maxi-anion channel as a candidate pathway for osmosensitive ATP release from mouse astrocytes in primary culture. AB - In the present study, we aimed to evaluate the pathways contributing to ATP release from mouse astrocytes during hypoosmotic stress. We first examined the expression of mRNAs for proteins constituting possible ATP-releasing pathways that have been suggested over the past several years. In RT-PCR analysis using both control and osmotically swollen astrocytes, amplification of cDNA fragments of expected size was seen for connexins (Cx32, Cx37, Cx43), pannexin 1 (Px1), the P2X7 receptor, MRP1 and MDR1, but not CFTR. Inhibitors of exocytotic vesicular release, gap junction hemi-channels, CFTR, MRP1, MDR1, the P2X7 receptor, and volume-sensitive outwardly rectifying chloride channels had no significant effects on the massive ATP release from astrocytes. In contrast, the hypotonicity induced ATP release from astrocytes was most effectively inhibited by gadolinium (50 muM), an inhibitor of the maxi-anion channel, which has recently been shown to serve as a pathway for ATP release from several other cell types. Thus, we propose that the maxi-anion channel constitutes a major pathway for swelling induced ATP release from cultured mouse astrocytes as well. PMID- 18414451 TI - Rheumatic fever: the potential advantages of technology. AB - This article sets out the clinical context of the research presented by Carapetis et al. in an accompanying article in this issue. No screening guidelines exist for early detection of rheumatic heart disease (RHD). The lack of agreed diagnostic criteria is one of the reasons why large proportions of cases in resource poor nations go untreated and are detected only when the disease has progressed to cardiac failure. Here, Poole-Wilson and Seth discuss the study carried out by Jonathan Carapetis and colleagues-a cross-sectional screening protocol in Tongan primary school children. The protocol designed by Carapetis et al. identified the highest echocardiographically confirmed prevalence of RHD reported to date. Poole-Wilson and Seth explore how modern technology, in the shape of portable echocardiography, can help to move the focus on rheumatic heart disease away from epidemiology and crude preventive programs, and towards screening programs that can effectively identify people who should receive treatment before cardiac failure occurs. PMID- 18414452 TI - Therapeutic targeting of the stem cell niche in experimental hindlimb ischemia. AB - BACKGROUND: The custom microenvironment 'vascular niche' is a potential therapeutic target for several pathophysiological conditions. Osteoblasts regulate the hematopoietic stem cell niche, and activation of the parathyroid hormone (PTH) receptor can increase the number of cells mobilized into the bloodstream. METHODS: C57Bl/6 mice were randomly assigned treatment with granulocyte-colony stimulating factor (G-CSF), PTH, G-CSF plus PTH or saline. All mice underwent hindlimb ischemia. Blood flow was measured by laser Doppler imaging. Indices of capillary activity were determined by electron microscopy in muscle tissue. CD34(+) and Ki67(+) cells were detected and evaluated by immunofluorescence, apoptosis by TUNEL, surface antigen and endothelial progenitor cells by fluorescence-activated cell sorting analysis, and vascular endothelial growth factor-164 and angiopoietin-1 expression by reverse transcriptase polymerase chain reaction. Frozen bone marrow sections were stained for antigen-specific B cells and fibronectin and analyzed by confocal laser scanning microscopy. RESULTS: Following mobilization induced by G-CSF treatment, mice also treated with PTH showed increases in blood flow, capillary density, nitrite/nitrate release, angiogenic factors and circulating progenitor cells, as well as reduced apoptosis, fibrosis, oxidative stress and inflammation in ischemic muscles. Furthermore, hematopoietic antigen-specific B cells in the bone marrow were also increased by G-CSF alone and in combination with PTH. CONCLUSIONS: PTH might increase the efficiency of hematopoietic stem-cell-based therapy in a recognized model of peripheral ischemia. Our translational experimental therapeutic targeting of the vascular niche points to novel clinical targets for the hematopoietic stem-cell treatment of ischemic vascular diseases. PMID- 18414453 TI - Safety of drug-eluting stents. AB - Drug-eluting stents (DESs) effectively reduce angiographic restenosis and the clinical need for repeat revascularization procedures as compared with bare-metal stents. Widely publicized concerns arose recently about the incidence of late and very late stent thrombosis with the use of first-generation DESs. Recent systematic reviews and large-scale registry studies demonstrated similar rates of overall mortality and myocardial infarction for patients treated with either DESs or bare-metal stents during long-term follow-up. Careful selection of stent type according to patient and lesion characteristics as well as monitoring of adherence to dual antiplatelet therapy could maximize the therapeutic potential of these devices. The purpose of the present Review is to provide the reader with an overview of the benefits and risks of first-generation DESs that could help physicians select the most appropriate stent type for each patient. PMID- 18414454 TI - Early intervention in acute coronary syndrome: is bivalirudin monotherapy an effective antithrombotic strategy? PMID- 18414455 TI - Management of boys with nonpalpable undescended testis. AB - Cryptorchidism is one of the most common genitourinary disorders in young boys. Although the management of boys with palpable testis is standardized, there are no formal guidelines for the management of boys with nonpalpable testis. In this Review we look at the current trends in the diagnosis and treatment of this disorder, as well as the indications for therapy and surgical procedures. On the basis of current evidence, we find that there is no optimum orchidopexy technique for the treatment of intra-abdominal testis, although it is preferable to adopt techniques that preserve the spermatic vessels. We also briefly examine the follow-up of patients with this disorder and its common complications. As yet, there are no data that assess the potential of laparoscopic orchidopexy being a risk factor for impaired fertility later in life. PMID- 18414456 TI - Is hypnotherapy effective for pediatric functional abdominal pain or IBS? PMID- 18414457 TI - Hepatitis B and C in children. AB - Chronic infection with hepatitis B affects nearly 350 million individuals worldwide and is the leading cause of hepatocellular carcinoma and liver cirrhosis. Universal infant immunization has decreased rates of HBV infection, although transmission continues to occur via vertical (mother-to-child) and horizontal (sexual, parenteral and household) routes. Treatments are now available for children with chronic HBV infection, but appropriate selection of those most likely to respond to treatment is important. Interferon alpha and lamivudine are currently approved in the US for the treatment of children older than 2 years of age who have chronic HBV infection. Hepatitis C infection affects almost 170 million individuals worldwide. Of individuals exposed to HCV, 60-80% develop chronic hepatitis, and 10-15% of those chronically infected develop cirrhosis within several decades. No vaccine exists for HCV; therefore, prevention of parenteral transmission is important. A high index of suspicion is essential for the diagnosis of HCV infection given its silent clinical presentation. Appropriate evaluation of infected individuals is warranted when considering their suitability for therapy. Interferon alpha and ribavirin, used in combination, are currently approved in the US for the treatment of children older than 3 years of age with chronic HCV infection. PMID- 18414458 TI - A case of undifferentiated connective tissue disease: is it a distinct clinical entity? AB - BACKGROUND: In November 2001, a 24-year-old woman with thrombocytopenia and Raynaud's phenomenon presented to our clinic. Her physical examination was unremarkable except for bruising on her legs and arms. INVESTIGATIONS: Laboratory assays detected the presence of antinuclear and anti-ribonucleoprotein antibodies. Tests for antibodies to double-stranded DNA and for antiphospholipid (lupus anticoagulant and anticardiolipin), anticentromere, anti-Scl-70, and antiplatelet antibodies were negative, as was a Coombs test. An echocardiogram, chest X-ray, and abdominal scan showed no abnormalities. Nailfold digital capillaroscopy revealed minor capillary changes not specific for scleroderma. DIAGNOSIS: Undifferentiated connective tissue disease. MANAGEMENT: The patient was successfully treated initially with high doses of corticosteroids and azathioprine, followed by rapid dose tapering. Therapy was continued for 2 years and then stopped. Over the next 4 years the patient's disease history was unremarkable, apart from mild Raynaud's phenomenon of the hands and the presence antinuclear and anti-ribonucleoprotein antibodies. The diagnosis of undifferentiated connective tissue disease was confirmed at her most-recent assessment, in October 2007. PMID- 18414459 TI - Drug insight: tumor necrosis factor-converting enzyme as a pharmaceutical target for rheumatoid arthritis. AB - The success of agents that inhibit tumor necrosis factor (TNF), such as infliximab, adalimumab and etanercept, has led to a desire for orally available small molecules that have a better safety profile and are less costly to produce than current agents. One target for anti-TNF therapy that is currently under investigation is TNF-converting enzyme, which promotes the release of soluble TNF from its membrane-bound precursor. Inhibitors of this enzyme with drug-like properties have been made and tested in the clinic. These inhibitors include TMI 005 and BMS-561392, both of which have entered into phase II clinical trials. This article summarizes preclinical and clinical findings regarding the use of inhibitors of TNF-converting enzyme for the treatment of rheumatoid arthritis. PMID- 18414460 TI - Use of intravenous sodium bicarbonate might increase the risk of contrast nephropathy. PMID- 18414461 TI - Breast cancer screening in women with chronic kidney disease: the unrecognized effects of metastatic soft-tissue calcification. AB - Patients with chronic kidney disease (CKD) or end-stage renal disease (ESRD) are known to develop metastatic soft-tissue calcification, secondary to hyperparathyroidism, in tissues including the breast. Such calcifications in women could pose a problem for interpretation of mammograms, since they are thought to mimic malignant lesions and interfere with differentiation of benign from malignant disease. Investigation of this issue is important to provide high quality, accurate breast care to women with CKD or ESRD, but little evidence is so far available. In a systematic review of the literature on the types and patterns of breast calcifications, we found only three studies that examined metastatic soft-tissue calcifications of the breast. The studies did, however, confirm that women with CKD or ESRD have a higher frequency of breast calcification than women with normal kidney function. The two older studies reported that these breast calcifications are not associated with malignancy, but the later study reported a raised rate of suspicious breast calcification among women with ESRD receiving hemodialysis, leading to an increased biopsy referral rate. In this Review we discuss the strengths and limitations of the available data and whether mammography is recommended in women with CKD or ESRD. PMID- 18414462 TI - What caused the decline in US breast cancer incidence? PMID- 18414463 TI - A patient with MEN1-associated hyperparathyroidism, responsive to cinacalcet. AB - BACKGROUND: A 30-year-old woman with suspected multiple endocrine neoplasia type 1 (MEN1) was referred to our center in 2001 with primary hyperparathyroidism caused by a multiglandular parathyroid adenoma. The patient also had hyperprolactinemia caused by an anterior pituitary macroadenoma. The patient underwent a parathyroidectomy with autotransplantation of parathyroid fragments into the nondominant forearm, resulting in resolution of the primary hyperparathyroidism. MEN1 was confirmed by analysis of the MEN1 gene, which revealed a 1555insG frameshift mutation. In 2006 serum calcium and parathyroid hormone (PTH) levels were again found to be high. INVESTIGATIONS: After parathyroidectomy in 2001, the patient underwent regular measurements of PTH levels from both forearms, of serum calcium, prolactin and phosphate levels, and of urinary calcium and phosphate levels. When serum calcium and PTH levels were found to be elevated in 2006, circulating PTH levels were similar in both forearms. Ultrasound scan and technetium-99m-labeled hexakis-2 methoxyisobutylisonitrile ((99m)Tc MIBI) scintigraphy evidenced a metabolically active parathyroid nodule in the neck. DIAGNOSIS: Local recurrence of a parathyroid adenoma associated with MEN1. MANAGEMENT: Because the patient refused a further operation, we decided to initiate pharmacological treatment with cinacalcet. After 1 month of therapy, serum calcium and PTH levels returned to normal. The patient has now been closely monitored for 1 year. During this time calcium and PTH levels remained normal, morphologically the parathyroid nodular lesion remained unchanged and cinacalcet was well tolerated without the occurrence of adverse events. Cinacalcet could represent an important pharmacological intervention in MEN1-associated primary hyperparathyroidism before surgery and in postsurgical recurrences. PMID- 18414464 TI - Treatment of impaired glucose tolerance in childhood. PMID- 18414465 TI - Diabetic nephropathy-an evaluation of potential risk factors in young patients with type 1 diabetes mellitus. PMID- 18414466 TI - Evidence for long-term IVIg treatment of chronic inflammatory demyelinating polyradiculoneuropathy. PMID- 18414467 TI - Do pilocytic astrocytomas have a benign course in adult patients? PMID- 18414468 TI - Hemiplegic cerebral palsy: role of repeat botulinum toxin A injections as an adjunct to occupational therapy. PMID- 18414469 TI - A dose-escalation study of indisulam in combination with capecitabine (Xeloda) in patients with solid tumours. AB - This dose escalation study was designed to determine the recommended dose of the multi-targeted cell cycle inhibitor indisulam in combination with capecitabine in patients with solid tumours and to evaluate the pharmacokinetics of the combination. Thirty-five patients were treated with indisulam on day 1 of each 21 day cycle. Capecitabine was administered two times daily (BID) on days 1-14. Plasma concentrations of indisulam, capecitabine and its three metabolites were determined for pharmacokinetic analysis. The main dose-limiting toxicity was myelosuppression. Hand/foot syndrome and stomatitis were the major non haematological toxicities. The recommended dose was initially established at indisulam 700 mg m(-2) and capecitabine 1250 mg m(-2) BID. However, during cycle 2 the recommended dose was poorly tolerated in three patients. A dose of indisulam 500 mg m(-2) and capecitabine 1250 mg m(-2) BID proved to be safe at cycle 1 and 2 in nine additional patients. Indisulam pharmacokinetics during cycle 1 were consistent with pharmacokinetic data from phase I mono-therapy studies. However, exposure to indisulam was remarkably increased at cycle 2 due to a drug-drug interaction between capecitabine and indisulam. Partial response was confirmed in two patients, one with colon carcinoma and the other with pancreatic carcinoma. Seventeen patients had stable disease. Indisulam (700 mg m( 2)) in combination with capecitabine (1250 mg m(-2) BID) was well tolerated during the first cycle. A dose of indisulam 500 mg m(-2) and capecitabine 1250 mg m(-2) BID was considered safe in multiple treatment cycles. The higher incidence of toxicities observed during cycle 2 can be explained by a time-dependent pharmacokinetic drug-drug interaction. PMID- 18414470 TI - Gene expression profiling may improve diagnosis in patients with carcinoma of unknown primary. AB - Carcinomas of unknown primary (CUP) represent between 3 and 10% of malignancies. Treatment with nonspecific chemotherapy is commonly unhelpful and the median survival is between 3 and 6 months. Gene expression microarray (GEM) analysis has demonstrated that molecular signatures can aid in tumour classification and propose foster primaries. In this study, we demonstrate the clinical utility of a diagnostic gene expression profiling tool and discuss its potential implications for patient management strategies. Paraffin tumour samples from 21 cases of 'true' CUP patients in whom standard investigation had failed to determine a primary site of malignancy were investigated using diagnostic gene profiling. The results were reviewed in the context of histology and clinical history. Classification of tumour origin using the GEM method confirmed the clinicians' suspicion in 16 out of 21 cases. There was a clinical/GEM inconsistency in 4 out of 21 patients and a pathological/GEM inconsistency in 1 patient. The improved diagnoses by the GEM method would have influenced the management in 12 out of 21 cases. Genomic profiling and cancer classification tools represent a promising analytical approach to assist with the management of CUP patients. We propose that GEM diagnosis be considered when the primary clinical algorithm has failed to provide a diagnosis. PMID- 18414471 TI - Reorganisation of Wnt-response pathways in colorectal tumorigenesis. AB - In most colorectal tumours, APC mutation stabilises beta-catenin and mimics elements of Wnt growth factor signalling, but the high frequency of epigenetic loss of Wnt antagonists indicates an additional role for ligand-mediated Wnt signalling. Here, we have investigated the expression of key components of beta catenin-independent Wnt response pathways to determine whether their profiles change during the transition from normal mucosa to colorectal adenomas. Transcription of the Wnt/planar cell polarity pathway determinant NKD1 (naked cuticle homologue 1) was induced in adenomas by a median 135-fold and in cancers by 7.4-fold. While some Frizzleds (FZDs) were downregulated in adenomas, the Wnt/Ca(2+) receptors FZD3 and FZD6 were induced by a median factor of 6.5 and 4.6, respectively. Naked cuticle homologue 1, FZD3 and FZD6 expression were coordinated in pre-malignant disease, but this relationship was lost in invasive cancers, where FZD induction was seen less frequently. Naked cuticle homologue 1 expression was associated with nuclear localisation of phospho-c-Jun in adenomas. In cultured cells, NKD1 transcription was induced by lithium chloride but FZD3 expression required Wnt growth factor treatment. These data show that Wnt responses are consistently directed towards both beta-catenin-independent routes in early colorectal tumorigenesis and elements of this are retained in more advanced cancers. These beta-catenin-independent Wnt signalling pathways may provide novel targets for chemoprevention of early colorectal tumours. PMID- 18414472 TI - Promoter hypermethylation of FANCF and outcome in advanced ovarian cancer. AB - The Fanconi gene family has a role in DNA repair and inactivation of FANCF has been proposed as a mechanism of sensitisation to platinum chemotherapy. This study sought to confirm this hypothesis in cell lines and a large series of ovarian cancer samples. Promoter methylation was assessed by methylation sensitive polymerase chain reaction of FANCF in nine ovarian cancer cell lines and 74 ovarian cancer samples taken from patients entered on a trial of cisplatin based chemotherapy. This study confirmed methylation-dependent silencing of FANCF in one out of nine ovarian cancer cell lines. Methylation of FANCF was demonstrated in 13.2% of 53 evaluable ovarian tumour samples. Progression-free survival gave an HR of 3.63 (95% CI: 1.54-8.54, P=0.0016) in favour of the unmethylated cases. There was no association with overall survival. This study does not support methylation-dependent silencing of FANCF as a mechanism of sensitisation to platinum-based chemotherapy in ovarian cancer. PMID- 18414473 TI - High level of telomerase RNA gene expression is associated with chromatin modification, the ALT phenotype and poor prognosis in liposarcoma. AB - Telomere length is maintained by two known mechanisms, activation of telomerase or alternative lengthening of telomeres (ALT). The ALT pathway is more commonly activated in tumours of mesenchymal origin, although the mechanisms involved in the decision of a cell to activate either telomerase or ALT are unknown at present and no molecular markers exist to define the ALT phenotype. We have previously shown an association between chromatin remodelling, telomerase gene expression and ALT in cell line models. Here, we evaluate these findings and investigate their prognostic significance in a panel of liposarcoma tissue samples to understand the biology underlying the ALT phenotype. Liposarcoma samples were split into three groups: telomerase positive (Tel+); ALT positive; ALT-/Tel-. Differences in telomerase gene expression were evident between the groups with increased expression of hTR in ALT and Tel+ compared to ALT-/Tel- samples and increased hTERT in Tel+ samples only. Investigation of a small panel of chromatin modifications revealed significantly increased binding of acetyl H3 in association with hTR expression. We confirm that the presence of the ALT phenotype is associated with poor prognosis and in addition, for the first time, we show a direct association between hTR expression and poor prognosis in liposarcoma patients. PMID- 18414474 TI - Early onset breast cancer in black British women: a letter to the editor of British Journal of Cancer regarding early onset of breast cancer in a group of British black women. PMID- 18414476 TI - Analysis of AAV serotypes 1-9 mediated gene expression and tropism in mice after systemic injection. AB - This study examines transgene expression and biodistribution of adeno-associated virus (AAV) pseudotyped 1-9 after tail vein (TV) injection in male mice. Using a cytomegalovirus (CMV)-luciferase transgene, the time-course of expression in each animal was tracked throughout the experiment. The animals were imaged at 7, 14, 29, 56, and 100 days after the TV injection. The total number of photons emitted from each animal was recorded, allowing examination of expression level and kinetics for each pseudotyped virus. The bioluminescence imaging revealed three expression levels (i) low-expression group, AAV2, 3, 4, and 5; (ii) moderate expression group, AAV1, 6, and 8; and (iii) high-expression group, AAV7 and 9. In addition, imaging revealed two classes of kinetics (i) rapid-onset, for AAV1, 6, 7, 8, and 9; and (ii) slow-onset, for AAV2, 3, 4, and 5. We next evaluated protein expression and viral genome copy numbers in dissected tissues. AAV9 had the best viral genome distribution and highest protein levels. The AAV7 protein and genome copy numbers were comparable to those of AAV9 in the liver. Most surprisingly, AAV4 showed the greatest number of genome copies in lung and kidney, and a high copy number in the heart. AAV6 expression was observed in the heart, liver, and skeletal muscle, and the genome distribution corroborated these observations. PMID- 18414477 TI - Systemic AAV6 delivery mediating RNA interference against SOD1: neuromuscular transduction does not alter disease progression in fALS mice. AB - Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disorder arising from the selective death of motor neurons. Approximately 20% of familial ALS (fALS) cases are caused by toxic gain-of-function mutations in the superoxide dismutase 1 (SOD1) gene. We as well as others have provided proof-of-principle for the use of RNA interference (RNAi) against mutant SOD1 as a potential therapy for fALS. With the aim of maximizing the delivery of these silencing instructions, we explored the efficacy of intravenous delivery of recombinant adeno-associated virus (rAAV) serotype 6 expressing small hairpin RNAs targeting mutant SOD1 in the G93A SOD1 fALS mouse model. This approach resulted in a systemic transduction profile, corresponding to transduction of the entire skeletal musculature as well as heart and liver. In addition, motor neurons at all levels of the spinal cord and brain stem were transduced, amounting to 3-5% of the lower motor neuron pool. SOD1 protein levels were reduced by >50% in all the muscles that were examined. Crucially, this silencing profile did not alter the course of the disease in this fALS model, thereby providing compelling evidence that SOD1-mediated damage within skeletal muscles does not contribute to death of motor neurons in ALS. Further, this study demonstrates that motor neurons can be transduced across the length of the spinal cord through a single noninvasive delivery of rAAV. PMID- 18414478 TI - Gene delivery to the juvenile mouse liver using AAV2/8 vectors. AB - Recombinant adeno-associated viral (rAAV) vectors have shown promise for use in liver-targeted gene delivery, but their effects have not been extensively investigated in the immature liver. Understanding the impact of liver growth on the efficacy of transduction is essential, because many monogenic liver diseases that are amenable to gene therapy will require treatment early in life. Here we show that rAAV2/8 transduces the neonatal mouse liver with high efficiency. With just one doubling in liver weight, however, there is a rapid reduction in vector genome numbers, irrespective of form, and the loss of episomal vector is almost complete by 2 weeks. Stable transgene expression is observed in a small percentage of hepatocytes, often in two- to eight-cell clusters, suggestive of genomic integration. Delivery at serially older ages was associated with progressively improved episome persistence and transgene expression. Vector re administration was possible following initial neonatal administration, albeit at reduced efficacy because of an anticapsid humoral immune response. We also found that intraperitoneal (i.p.) delivery of rAAV2/8 was highly effective at all ages, and that promoter selection is the critical determinant of the intensity and pattern of transgene expression across the hepatic lobule. We conclude that successful use of rAAV to treat liver disease in early childhood will require optimally efficient vector constructs and probable re-administration. PMID- 18414479 TI - Treatment of obesity and diabetes in mice by transplant of gut cells engineered to produce leptin. AB - Leptin injections evoke weight loss by causing a reduction in food consumption and an increase in energy expenditure. Also, the administration of leptin lowers blood glucose levels in some rodent models of diabetes and in humans with lipodystrophy. We explored the therapeutic potential of delivering leptin to obese, diabetic ob/ob mice and to mice fed on a high-fat diet (HFD), by transplanting gut-derived cells engineered to produce leptin, under the regulation of an inducing agent, mifepristone. These cells expressed and released leptin in a mifepristone dose-dependent and time-dependent manner. The engineered cells were either transplanted into the mice under the kidney capsule or were encapsulated in alginate and injected into the intraperitoneal cavity, while mifepristone was delivered by implanting 14-day release pellets. In ob/ob mice, leptin delivery by this method caused a significant reduction in food intake and profound weight loss, which was controllable by adjusting the dose of mifepristone. These transplants also achieved rapid and persistent amelioration of diabetes. However, mice fed on a HFD were resistant to the leptin therapy. These results indicate that gut cells can be modified to express leptin in an inducible manner and that the transplantation of these cells has a therapeutic effect in leptin-deficient mice, but not in mice fed on a HFD. PMID- 18414480 TI - Emergent dilated cardiomyopathy caused by targeted repair of dystrophic skeletal muscle. AB - Duchenne muscular dystrophy (DMD) is a fatal disease characterized by deterioration of striated muscle, affecting skeletal and cardiac muscles. Recently, several therapeutic approaches have shown promise for repairing dystrophic skeletal muscles. However, these methods often leave the dystrophic heart untreated. Here we show that, in comparison to fully dystrophin-deficient animals, targeted transgenic repair of skeletal muscle, but not cardiac muscle, in otherwise dystrophin-deficient (mdx) mice paradoxically elicited a fivefold increase in cardiac injury and dilated cardiomyopathy in these animals in vivo. Skeletal muscle repair was shown to increase the voluntary activity of the mdx mice as quantified by voluntary running on the exercise wheel. Because the dystrophin-deficient heart is highly sensitive to increased stress, we hypothesize that increased activity (enabled by the repaired skeletal muscle) provided the stimulus for heightened cardiac injury and heart remodeling. In support of this hypothesis, the primary cellular compliance defect in dystrophin deficient cardiac myocytes was found to be unchanged by skeletal muscle repair in the mdx mice. These findings provide new information on the evolution of cardiac disease in dystrophin-deficient animals and underscore the importance of implementing global striated muscle therapies for muscular dystrophy. PMID- 18414481 TI - A map of human protein interactions derived from co-expression of human mRNAs and their orthologs. AB - The human protein interaction network will offer global insights into the molecular organization of cells and provide a framework for modeling human disease, but the network's large scale demands new approaches. We report a set of 7000 physical associations among human proteins inferred from indirect evidence: the comparison of human mRNA co-expression patterns with those of orthologous genes in five other eukaryotes, which we demonstrate identifies proteins in the same physical complexes. To evaluate the accuracy of the predicted physical associations, we apply quantitative mass spectrometry shotgun proteomics to measure elution profiles of 3013 human proteins during native biochemical fractionation, demonstrating systematically that putative interaction partners tend to co-sediment. We further validate uncharacterized proteins implicated by the associations in ribosome biogenesis, including WBSCR20C, associated with Williams-Beuren syndrome. This meta-analysis therefore exploits non-protein-based data, but successfully predicts associations, including 5589 novel human physical protein associations, with measured accuracies of 54+/-10%, comparable to direct large-scale interaction assays. The new associations' derivation from conserved in vivo phenomena argues strongly for their biological relevance. PMID- 18414482 TI - Synthetic gene brushes: a structure-function relationship. AB - We present the assembly of gene brushes by means of a photolithographic approach that allows us to control the density of end-immobilized linear double-stranded DNA polymers coding for entire genes. For 2 kbp DNAs, the mean distance varies from 300 nm, where DNAs are dilute and assume relaxed conformations, down to 30 nm, where steric repulsion at dense packing forces stretching out. We investigated the gene-to-protein relationship of firefly luciferase under the T7/E.Coli-extract expression system, as well as transcription-only reactions with T7 RNA polymerase, and found both systems to be highly sensitive to brush density, conformation, and orientation. A 'structure-function' picture emerges in which extension of genes induced by moderate packing exposes coding sequences and improves their interaction with the transcription/translation machinery. However, tighter packing impairs the penetration of the machinery into the brush. The response of expression to two-dimensional gene crowding at the nanoscale identifies gene brushes as basic controllable units en route to multicomponent synthetic systems. In turn, these brushes could deepen our understanding of biochemical reactions taking place under confinement and molecular crowding in living cells. PMID- 18414483 TI - Ultrasensitive gene regulation by positive feedback loops in nucleosome modification. AB - Eukaryotic transcription involves the synergistic interaction of many different proteins. However, the question remains how eukaryotic promoters achieve ultrasensitive or threshold responses to changes in the concentration or activity of a single transcription factor (TF). We show theoretically that by recruiting a histone-modifying enzyme, a TF binding non-cooperatively to a single site can change the balance between opposing positive feedback loops in histone modification to produce a large change in gene expression in response to a small change in concentration of the TF. This mechanism can also generate bistable promoter responses, allowing a gene to be on in some cells and off in others, despite the cells being in identical conditions. In addition, the system provides a simple means by which the activities of many TFs could be integrated at a promoter. PMID- 18414484 TI - Mathematical modeling of pathogenicity of Cryptococcus neoformans. AB - Cryptococcus neoformans (Cn) is the most common cause of fungal meningitis worldwide. In infected patients, growth of the fungus can occur within the phagolysosome of phagocytic cells, especially in non-activated macrophages of immunocompromised subjects. Since this environment is characteristically acidic, Cn must adapt to low pH to survive and efficiently cause disease. In the present work, we designed, tested, and experimentally validated a theoretical model of the sphingolipid biochemical pathway in Cn under acidic conditions. Simulations of metabolic fluxes and enzyme deletions or downregulation led to predictions that show good agreement with experimental results generated post hoc and reconcile intuitively puzzling results. This study demonstrates how biochemical modeling can yield testable predictions and aid our understanding of fungal pathogenesis through the design and computational simulation of hypothetical experiments. PMID- 18414485 TI - Transient heterogeneity in extracellular protease production by Bacillus subtilis. AB - The most sophisticated survival strategy Bacillus subtilis employs is the differentiation of a subpopulation of cells into highly resistant endospores. To examine the expression patterns of non-sporulating cells within heterogeneous populations, we used buoyant density centrifugation to separate vegetative cells from endospore-containing cells and compared the transcriptome profiles of both subpopulations. This demonstrated the differential expression of various regulons. Subsequent single-cell analyses using promoter-gfp fusions confirmed our microarray results. Surprisingly, only part of the vegetative subpopulation highly and transiently expresses genes encoding the extracellular proteases Bpr (bacillopeptidase) and AprE (subtilisin), both of which are under the control of the DegU transcriptional regulator. As these proteases and their degradation products freely diffuse within the liquid growth medium, all cells within the clonal population are expected to benefit from their activities, suggesting that B. subtilis employs cooperative or even altruistic behavior. To unravel the mechanisms by which protease production heterogeneity within the non-sporulating subpopulation is established, we performed a series of genetic experiments combined with mathematical modeling. Simulations with our model yield valuable insights into how population heterogeneity may arise by the relatively long and variable response times within the DegU autoactivating pathway. PMID- 18414486 TI - Systems engineering to systems biology. PMID- 18414488 TI - A synthetic Escherichia coli predator-prey ecosystem. AB - We have constructed a synthetic ecosystem consisting of two Escherichia coli populations, which communicate bi-directionally through quorum sensing and regulate each other's gene expression and survival via engineered gene circuits. Our synthetic ecosystem resembles canonical predator-prey systems in terms of logic and dynamics. The predator cells kill the prey by inducing expression of a killer protein in the prey, while the prey rescue the predators by eliciting expression of an antidote protein in the predator. Extinction, coexistence and oscillatory dynamics of the predator and prey populations are possible depending on the operating conditions as experimentally validated by long-term culturing of the system in microchemostats. A simple mathematical model is developed to capture these system dynamics. Coherent interplay between experiments and mathematical analysis enables exploration of the dynamics of interacting populations in a predictable manner. PMID- 18414489 TI - Genomic analysis of estrogen cascade reveals histone variant H2A.Z associated with breast cancer progression. AB - We demonstrate an integrated approach to the study of a transcriptional regulatory cascade involved in the progression of breast cancer and we identify a protein associated with disease progression. Using chromatin immunoprecipitation and genome tiling arrays, whole genome mapping of transcription factor-binding sites was combined with gene expression profiling to identify genes involved in the proliferative response to estrogen (E2). Using RNA interference, selected ERalpha and c-MYC gene targets were knocked down to identify mediators of E2 stimulated cell proliferation. Tissue microarray screening revealed that high expression of an epigenetic factor, the E2-inducible histone variant H2A.Z, is significantly associated with lymph node metastasis and decreased breast cancer survival. Detection of H2A.Z levels independently increased the prognostic power of biomarkers currently in clinical use. This integrated approach has accelerated the identification of a molecule linked to breast cancer progression, has implications for diagnostic and therapeutic interventions, and can be applied to a wide range of cancers. PMID- 18414490 TI - Non-hexameric DNA helicases and translocases: mechanisms and regulation. AB - Helicases and nucleic acid translocases are motor proteins that have essential roles in nearly all aspects of nucleic acid metabolism, ranging from DNA replication to chromatin remodelling. Fuelled by the binding and hydrolysis of nucleoside triphosphates, helicases move along nucleic acid filaments and separate double-stranded DNA into their complementary single strands. Recent evidence indicates that the ability to simply translocate along single-stranded DNA is, in many cases, insufficient for helicase activity. For some of these enzymes, self assembly and/or interactions with accessory proteins seem to regulate their translocase and helicase activities. PMID- 18414491 TI - Expansion and evolution of cell death programmes. AB - Cell death has historically been subdivided into regulated and unregulated mechanisms. Apoptosis, a form of regulated cell death, reflects a cell's decision to die in response to cues and is executed by intrinsic cellular machinery. Unregulated cell death (often called necrosis) is caused by overwhelming stress that is incompatible with cell survival. Emerging evidence, however, suggests that these two processes do not adequately explain the various cell death mechanisms. Recent data point to the existence of multiple non-apoptotic, regulated cell death mechanisms, some of which overlap or are mutually exclusive with apoptosis. Here we examine how and why these different cell death programmes have evolved, with an eye towards new cytoprotective therapeutic opportunities. PMID- 18414492 TI - A kinder, gentler way: control of the proliferative tumor compartment, not cosmetic complete response, should be the goal of myeloma therapy. PMID- 18414493 TI - C/EBPalpha, do not forget your TIP60. PMID- 18414495 TI - Neural progenitor cells treated with EPO induce angiogenesis through the production of VEGF. AB - Recombinant human erythropoietin (rhEPO) induces neurogenesis and angiogenesis. Using a coculture system of mouse brain endothelial cells (MBECs) and neural progenitor cells derived from the subventricular zone of adult mouse, we investigated the hypothesis that neural progenitor cells treated with rhEPO promote angiogenesis. Treatment of neural progenitor cells with rhEPO significantly increased their expression and secretion of vascular endothelial growth factor (VEGF) and activated phosphatidylinositol 3-kinase/Akt (PI3K/Akt) and extracellular signal-regulated kinase (ERK1/2). Selective inhibition of the Akt and ERK1/2 signaling pathways significantly attenuated the rhEPO-induced VEGF expression in neural progenitor cells. The supernatant harvested from neural progenitor cells treated with rhEPO significantly increased the capillary-like tube formation of MBECs. SU1498, a specific VEGF type-2 receptor (VEGFR2) antagonist, abolished the supernatant-enhanced angiogenesis. In addition, coculture of MBECs with neural progenitor cells treated with rhEPO substantially increased VEGFR2 mRNA and protein levels in MBECs. These in vitro results suggest that EPO enhances VEGF secretion in neural progenitor cells through activation of the PI3K/Akt and ERK1/2 signaling pathways and that neural progenitor cells treated with rhEPO upregulate VEGFR2 expression in cerebral endothelial cells, which along with VEGF secreted by neural progenitor cells promotes angiogenesis. PMID- 18414497 TI - Role of cortical spreading depressions for secondary brain damage after traumatic brain injury in mice. AB - In recent years, several studies have unequivocally shown the occurrence of cortical spreading depressions (CSDs) after stroke and traumatic brain injury (TBI) in humans. The fundamental question, however, is whether CSDs cause or result from secondary brain damage. The aim of the current study was, therefore, to investigate the role of CSDs for secondary brain damage in an experimental model of TBI. C57/BL6 mice were traumatized by controlled cortical impact. Immediately after trauma, each animal showed one heterogeneous direct current (DC) potential shift accompanied by a profound depression of electroencephalogram (EEG) amplitude, and a temporary decrease of ipsi- and contralateral regional cerebral blood flow (rCBF) suggesting bilateral CSDs. Within the next 3 h after TBI, CSDs occurred at a low frequency (0.38 CSD/h per animal, n=7) and were accompanied by rCBF changes confined to the ipsilateral hemisphere. No significant relationship between the number of SDs and lesion size or intracranial pressure (ICP) could be detected. Even increasing the number of posttraumatic CSDs by application of KCl by more than six times did not increase ICP or contusion volume. We therefore conclude that CSDs may not contribute to posttraumatic secondary brain damage in the normally perfused and oxygenated brain. PMID- 18414496 TI - Antiangiogenic effect of inhibitors of cytochrome P450 on rats with glioblastoma multiforme. AB - Cytochrome P450 epoxygenase catalyzes 5,6-, 8,9-, 11,12-, and 14,15 epoxyeicosatrienoic acids (EETs) from arachidonic acid (AA). In 1996, our group identified the expression of the cytochrome P450 2C11 epoxygenase (CYP epoxygenase) gene in astrocytes. Because of our finding an array of physiological functions have been attributed to EETs in the brain, one of the actions of EETs involves a predominant role in brain angiogenesis. Blockade of EETs formation with different epoxygenase inhibitors decreases endothelial tube formation in cocultures of astrocytes and capillary endothelial cells. The intent of this investigation was to determine if pharmacologic inhibition of formation of EETs is effective in reducing capillary formation in glioblastoma multiforme with a concomitant reduction in tumor volume and increase in animal survival time. Two mechanistically different inhibitors of CYP epoxygenase, 17-octadecynoic acid (17 ODYA) and miconazole, significantly reduced capillary formation and tumor size in glial tumors formed by injection of rat glioma 2 (RG2) cells, also resulting in an increased animal survival time. However, we observed that 17-ODYA and miconazole did not inhibit the formation of EETs in tumor tissue. This implies that 17-ODYA and miconazole appear to exert their antitumorogenic function by a different mechanism that needs to be explored. PMID- 18414498 TI - Preischemic induction of TNF-alpha by physical exercise reduces blood-brain barrier dysfunction in stroke. AB - This study explores the neuroprotective action of tumor necrosis factor-alpha (TNF-alpha) induced during physical exercise, which, consequently, reduces matrix metalloproteinase-9 (MMP-9) activity and ameliorates blood-brain barrier (BBB) dysfunction in association with extracellular signal-regulated kinase 1 and 2 (ERK1/2) phosphorylation. Adult male Sprague-Dawley rats were subjected to exercise on a treadmill for 3 weeks. A 2-h middle cerebral artery occlusion and reperfusion was administered to exercised and nonexercised animals to induce stroke. Exercised ischemic rats were subjected to TNF-alpha inhibition and ERK1/2 by TNF-alpha antibody or UO126. Nissl staining of coronal sections revealed the infarct volume. Evans blue extravasation and water content evaluated BBB function. Western blot was performed to analyze protein expression of TNF-alpha, ERK1/2, phosphorylated ERK1/2, the basal laminar protein collagen IV, and MMP-9. The activity of MMP-9 was determined by gelatin zymography. Tumor necrosis factor alpha expression and ERK1/2 phosphorylation were upregulated during exercise. Infarct volume, brain edema, and Evans blue extravasation all significantly decreased in exercised ischemic rats. Collagen IV production increased in exercised rats and remained high after stroke, whereas MMP-9 protein level and activity decreased. These results were negated and returned toward nonexercised values once TNF-alpha or ERK1/2 was blocked. We concluded that preischemic, exercise-induced TNF-alpha markedly decreases BBB dysfunction by using the ERK1/2 pathway. PMID- 18414499 TI - Incensole acetate: a novel neuroprotective agent isolated from Boswellia carterii. AB - Boswellia resin has been used as a major anti-inflammatory agent and for the healing of wounds for centuries. Incensole acetate (IA), isolated from this resin, was shown to inhibit the activation of nuclear factor-kappaB, a key transcription factor in the inflammatory response. We now show that IA inhibits the production of inflammatory mediators in an in vitro model system of C6 glioma and human peripheral monocytes. Given the involvement of postinjury inflammation in the pathophysiology and outcome of traumatic brain injury, we examined the effect of IA on the inflammatory process and on the recovery of neurobehavioral and cognitive functions in a mouse model of closed head injury (CHI). In the brains of post-CHI mice, IA reduced glial activation, inhibited the expression of interleukin-1beta, and tumor necrosis factor-alpha mRNAs, and induced cell death in macrophages at the area of trauma. A mild hypothermic effect was also noted. Subsequently, IA inhibited hippocampal neurodegeneration and exerted a beneficial effect on functional outcome after CHI, indicated by reduced neurological severity scores and improved cognitive ability in an object recognition test. This study attributes the anti-inflammatory activity of Boswellia resin to IA and related cembranoid diterpenes and suggests that they may serve as novel neuroprotective agents. PMID- 18414500 TI - Single-cell identification in microbial communities by improved fluorescence in situ hybridization techniques. AB - The ribosomal-RNA (rRNA) approach to microbial evolution and ecology has become an integral part of environmental microbiology. Based on the patchy conservation of rRNA, oligonucleotide probes can be designed with specificities that range from the species level to the level of phyla or even domains. When these probes are labelled with fluorescent dyes or the enzyme horseradish peroxidase, they can be used to identify single microbial cells directly by fluorescence in situ hybridization. In this Review, we provide an update on the recent methodological improvements that have allowed more reliable quantification of microbial populations in situ in complex environmental samples, with a particular focus on the usefulness of group-specific probes in this era of ever-growing rRNA databases. PMID- 18414501 TI - Modification of intracellular membrane structures for virus replication. AB - Viruses are intracellular parasites that use the host cell they infect to produce new infectious progeny. Distinct steps of the virus life cycle occur in association with the cytoskeleton or cytoplasmic membranes, which are often modified during infection. Plus-stranded RNA viruses induce membrane proliferations that support the replication of their genomes. Similarly, cytoplasmic replication of some DNA viruses occurs in association with modified cellular membranes. We describe how viruses modify intracellular membranes, highlight similarities between the structures that are induced by viruses of different families and discuss how these structures could be formed. PMID- 18414502 TI - Emerging and re-emerging rickettsioses: endothelial cell infection and early disease events. AB - Rickettsiae cause some of the most severe human infections, including epidemic typhus and Rocky Mountain spotted fever. Substantial progress has been made in research into the genomics, vector relationships, pathogenesis and immunity of these obligate, intracellular, arthropod-transmitted bacteria. This Review summarizes our understanding of the early and late events in pathogenesis and immunity, modulation of the host response to rickettsial infection by the vector, host defence, virulence mechanisms and rickettsial manipulation of host cells. PMID- 18414503 TI - Making a difference: 30 years of TDR. AB - When the UNICEF/UNDP/World Bank/WHO Special Programme for Research and Training in Tropical Diseases (TDR) was established in the mid-1970s, it represented an innovative institutional formula in terms of its structure and the manner in which scientists were convened from both developed and developing countries to address some of the world's most neglected parasitic diseases. A review of TDR's historical record sheds light not only on some important milestones in tropical disease research, but also on how future challenges could be approached and hopefully surmounted. PMID- 18414504 TI - Components of variation in serum carotenoid concentrations: the Polyp Prevention Trial. AB - OBJECTIVES: The intra- and interindividual variations and season and center effects were estimated from a series of serum carotenoid concentrations in the Polyp Prevention Trial (PPT) participants. SUBJECTS/METHODS: Fasting blood was collected annually for 4 years in all 1905 participants, and a subcohort of 901 participants were selected within each (of eight) center(s), by gender and dietary arm of the study, for measurement of five major carotenoid peaks. Using variance of component methods, the variation in serum carotenoid concentrations about the underlying mean was partitioned into explanatory components attributed to various sources. RESULTS: The contributions of the inter- and intraindividual variances to the overall variation in carotenoid concentrations were in the range of 61-70 and 20-35%, respectively, whereas center and center-by-season effects provided 2.6-9.5 and 0.2-1.4%, respectively. The highest percent (35%) of intraindividual variation was exhibited by lycopene, and the highest percent (70% apiece) of interindividual variation was exhibited by lutein/zeaxanthin and beta carotene. Serum lycopene had the highest ratio of intra- to interindividual variation of 0.57, whereas lutein had the lowest ratio of 0.29. We estimate that the ratio of intra- to interindividual variance around the mean carotenoid concentration can be reduced greatly by collecting 3-4 compared to 1 blood measurement in large-scale trials like the PPT. CONCLUSION: In the largest study of components of variation in individuals at high risk for colorectal cancer, the largest contributors to variation in serum carotenoid concentrations were intra- and interindividual effects followed by center and center-by-season effects. PMID- 18414505 TI - Satellite DNA in insects: a review. AB - The study of insect satellite DNAs (satDNAs) indicates the evolutionary conservation of certain features despite their sequence heterogeneity. Such features can include total length, monomer length, motifs, particular regions and/or secondary and tertiary structures. satDNAs may act as protein-binding sites, structural domains or sites for epigenetic modifications. The selective constraints in the evolution of satDNAs may be due to the satDNA sequence interaction with specific proteins important in heterochromatin formation and possible a role in controlling gene expression. The transcription of satDNA has been described in vertebrates, invertebrates and plants. In insects, differential satDNA expression has been observed in different cells, developmental stages, sex and caste of the individuals. These transcription differences may suggest their involvement in gene-regulation processes. In addition, the satDNA or its transcripts appear to be involved in heterochromatin formation and in chromatin elimination processes. The importance of transposable elements to insect satDNA is shown by their presence as a constituent of satDNA in several species of insects (including possible active elements). In addition, they may be involved in the formation of centromeres and telomeres and in the homogenization and expansion of satDNA. PMID- 18414506 TI - Variation in mate preference across a house mouse hybrid zone. AB - Reproductive character displacement is known to occur at the borders of a secondary contact zone between the two European subspecies of the house mouse (in Jutland, Denmark), where selection against hybridization occurs. This study assessed patterns of mate preference in naturally occurring hybrids of the two subspecies. Mate odour choice was investigated in male and female mice sampled across the hybrid zone. Odour samples comprised urine (from the opposite sex to the test animal) obtained from populations geographically distant from the hybrid zone. Urine is known to carry subspecies recognition signals. The behavioural results changed across the hybrid zone, and were analysed by a model of clinal variation. This behavioural cline was compared with the allozyme cline across the same hybrid zone. Males on both sides of the hybrid zone showed an assortative preference, which shifted significantly and abruptly approximately 10 km from the genetic centre of the hybrid zone on the Mus musculus musculus side. Directional preference was not detected in females, which could relate to variation in sexual receptivity. Our model indicates that the peculiar pattern of male preference could involve several genes and be characterized by mild to strong epistasis favouring the expression of M. m. domesticus-like preference over a large portion of the hybrid zone. This study may provide the first picture of the genetic determination of mate preference in a mammal. PMID- 18414507 TI - Identification of entire LMX1B gene deletions in nail patella syndrome: evidence for haploinsufficiency as the main pathogenic mechanism underlying dominant inheritance in man. AB - Heterozygous mutations in the LMX1B gene cause nail patella syndrome (NPS) that is associated with nail and skeletal malformations, nephropathy, and glaucoma. Previous phenotype studies of Lmx1b null mice revealed dorsal limb and renal anomalies similar to human NPS, which contributed to the identification of heterozygous mutations in this LIM-homeodomain protein LMX1B as the genetic defect responsible for NPS. Despite advanced insight into the role of the Lmx1b transcription factor in a broad range of animal developmental programs, the pathogenic mechanism underlying dominant inheritance of NPS in man remained unclear. Here, we describe for the first time the detection of two entire LMX1B gene deletions and one smaller exonic LMX1B deletion by multiplex ligation dependent probe amplification (MLPA) in a series of eight unrelated families with classical features of NPS in whom no pathogenic LMX1B mutation was found by sequence analysis. The identification of entire LMX1B deletions strongly confirms that haploinsufficiency is the principal pathogenetic mechanism of NPS and suggests a difference in dosage sensitivity for this gene between mice and man. PMID- 18414508 TI - Frequent mutations in the 3'-untranslated region of IFNGR1 lack functional impairment in microsatellite-unstable colorectal tumours. AB - Microsatellite repeats are frequently found to be mutated in microsatellite instable colorectal tumours. This suggests that these mutations are important events during tumour development. We have observed frequent mutations in microsatellite-instable (MSI-H) tumours and cell lines of a conserved A14 repeat within the 3'-untranslated region of the interferon-gamma receptor 1 gene (IFNGR1). The repeat was mutated in 59% (41 of 70) of colon carcinomas and in all four MSI-H colon cancer cell lines tested. In-vitro analysis of these cell lines did not show a decreased responsiveness to standard IFNgamma concentrations when compared to microsatellite-stable colon cancer cell lines. A functional consequence of the frequently found microsatellite instability in IFNGR1 is therefore not evident. PMID- 18414509 TI - The polymorphisms of protein-tyrosine phosphatase receptor-type delta gene and its association with pediatric asthma in the Taiwanese population. AB - We previously reported an association between genetic differences of pediatric asthma subtypes and a short tandem repeat (STR) marker, D9S286. It has been known that the protein-tyrosine phosphatase receptor-type delta (PTPRD) gene is located downstream of D9S286 and that the physical distance between them is about 0.25 Mb. We selected and conducted genotyping on 76 single-nucleotide polymorphisms (SNPs) that encircle the genomic region of PTPRD in Taiwanese children with or without asthma. A total of 996 subjects were divided into testing group (674 subjects) and validation group (322 subjects). The results were further validated with the third subject group (611 subjects) recruited from different geographical regions. After Bonferroni correction, 3 out of 80 SNPs were found to be strongly significant (P < 0.05/76 = 0.000658) in the allele frequency test. This association was confirmed by validation groups. The results indicate that polymorphisms of PTPRD are strongly associated with pediatric bronchial asthma in the Taiwanese population. PMID- 18414510 TI - Confirmation of the genetic association between the U2AF homology motif (UHM) kinase 1 (UHMK1) gene and schizophrenia on chromosome 1q23.3. AB - UHMK1 has previously been implicated as a susceptibility gene for schizophrenia in the 1q23.3 region by significant evidence of allelic and haplotypic association between schizophrenia and several genetic markers at UHMK1 in a London-based case-control sample. Further fine mapping of the UHMK1 gene locus in the University College London schizophrenia case-control sample was carried out with tagging SNPs. Two additional SNPs were found to be associated with schizophrenia (rs6604863 P = 0.02, rs10753578 P = 0.017). Tests of allelic and haplotypic association were then carried out in a second independent sample from Aberdeen consisting of 858 individuals with schizophrenia and 591 controls. Two of these SNPs also showed association in the Aberdeen sample (rs7513662 P = 0.0087, rs10753578 P = 0.022) and several haplotypes were associated (global permutation P = 0.0004). When the UCL and Aberdeen samples were combined three SNPs (rs7513662 P = 0.0007, rs6427680 P = 0.0252, rs6694863 P = 0.015) and several haplotypes showed association (eg HAP-A, HAP-B, HAP-C permutation P = 0.00005). The finding of allelic association with markers in the UHMK1 gene might help explain why it has not been possible, despite great effort, to satisfactorily confirm previously reported associations between schizophrenia and the genes RGS4 and NOS1AP/CAPON. These genes flank UHMK1 and all three loci are within a 700 kb region showing linkage to schizophrenia. The confirmation of association between UHMK1 and schizophrenia, rather than RGS4 and NOS1AP in the London sample, points to the possibility that previous efforts to accurately fine map a gene in the 1q23.3 region have lacked accuracy or may have suffered from methodological flaws. PMID- 18414511 TI - The effect of pedigree structure on detection of deletions and other null alleles. AB - Deletions and other null alleles for genetic markers can be detected as a special case of non-Mendelian inheritance, ie when a parent and a child appear to be homozygous for different alleles. The probability to detect a deletion for a fixed overall number of investigated individuals was calculated for biallelic and multiallelic markers with varying allele frequencies. To determine the effect of increasing the number of parents and grandparents, the probability for this event was derived for a parent and one child, a trio, a trio with one grandparent and a trio with two grandparents. The results for biallelic markers show that for a fixed total number of individuals, a sample of trios with two grandparents is always more efficient than the other family types, despite a lower total number of founder chromosomes in the sample. For multiallelic markers the outcome varies. The effect of adding additional children to a nuclear family was also investigated. For nuclear families, the optimal number of children is two or three, depending on the allele frequencies. It is shown that adding children is more efficient than adding grandparents. PMID- 18414512 TI - Position effect leading to haploinsufficiency in a mosaic ring chromosome 14 in a boy with autism. AB - We describe an individual with autism and a coloboma of the eye carrying a mosaicism for a ring chromosome consisting of an inverted duplication of proximal chromosome 14. Of interest, the ring formation was associated with silencing of the amisyn gene present in two copies on the ring chromosome and located at 300 kb from the breakpoint. This observation lends further support for a locus for autism on proximal chromosome 14. Moreover, this case suggests that position effects need to be taken into account, when analyzing genotype-phenotype correlations based on chromosomal imbalances. PMID- 18414513 TI - Use of a simple pharmacokinetic model to characterize exposure to perchlorate. AB - A simple two-compartment first-order pharmacokinetic model that predicts concentrations of perchlorate in blood and urine was constructed and validated. The model was validated using data from a high-dose experiment in humans where doses and resulting concentrations of perchlorate in blood and urine were well documented. Specifically, data were available for individuals who had been dosed at 0.5, 0.1, and 0.02 mg/kg/day for 14 consecutive days, significantly higher than the average background dose, which is estimated to be less than 0.0001 mg/kg/day. The average measured urine concentration in the high-dose regime during the experiment was 15.4 mg/l compared with an average prediction of 17.3 mg/l. In the medium-dose regime, the average measured was 3.0 mg/l compared with 4.1 mg/l predicted, and in the low-dose regime, the average measured was 0.53 mg/l compared with 0.68 mg/l predicted. For blood, the analogous results include 0.51 mg/l measured compared with 0.54 mg/l predicted in the high-dose regime and 0.12 mg/l measured versus 0.11 mg/l predicted in the medium-dose regime. The model was then used to study background exposures to perchlorate. A national sampling of perchlorate in urine showed a median concentration of 0.0035 mg/l, and this was used to back-calculate a dose of 0.000064 mg/kg/day. This finding was independently verified with the modeling structure of this study, as use of that back-calculated dose of 0.000064 mg/kg/day resulted in predictions of urine concentration with an average virtually identical at 0.0033 mg/l. An examination of literature data on the possible pathways of exposure suggests that the consumption of foods, rather than ingestion of water, dominates background exposures. Daily variation in urine concentration was studied with the model, and it was found that concentrations in the morning hours were lower than concentrations in the afternoon and evening hours, corresponding to the time when most exposure was assumed to occur. PMID- 18414514 TI - Surface-to-food pesticide transfer as a function of moisture and fat content. AB - Transfer of pesticides from household surfaces to foods may result in excess dietary exposure in children (i.e., beyond that inherent in foods due to agricultural application). In this study, transfer was evaluated as a function of the moisture and fat content of various foods. Surfaces chosen for investigation were those commonly found in homes and included Formica, ceramic tile, plastic, carpet, and upholstery fabric. Each surface type was sprayed with an aqueous emulsion of organophosphates, fipronil, and synthetic pyrethroids. In the first phase of the study, multiple foods (apples, watermelon, wheat crackers, graham crackers, white bread, flour tortillas, bologna, fat-free bologna, sugar cookies, ham, Fruit Roll-ups, pancakes, and processed American cheese) were categorized with respect to moisture and fat content. All were evaluated for potential removal of applied pesticides from a Formica surface. In the second phase of the study, representative foods from each classification were investigated for their potential for pesticide transfer with an additional four surfaces: ceramic tile, plastic, upholstery, and carpet. Moisture content, not fat, was found to be a determining factor in most transfers. For nearly all surfaces, more efficient transfer occurred with increased hardness (Formica and ceramic tile). Comparatively, the polymer composition of the plastic delivered overall lower transfer efficiencies, presumably due to an attraction between it and the organic pesticides of interest. PMID- 18414515 TI - Bisphenol A (BPA) daily intakes in the United States: estimates from the 2003 2004 NHANES urinary BPA data. AB - Investigations into human exposure to bisphenol A (BPA) have, for the most part, assessed intake based on food consumption estimates combined with measurements or estimates of BPA in foods. In this study, nationally representative data on urinary levels of BPA in the United States (US) from the 2003-2004 National Health and Nutrition Examination Survey (NHANES) were used to estimate daily intake of BPA, assuming steady-state excretion. Distributions of intakes for the US population were determined for (i) all NHANES participants with urinary BPA data; (ii) participants by the following age groups: 6-11 years, 12-19 years, 20 39 years, 40-59 years, and 60+ years; and (iii) participants by gender. On the basis of the NHANES urinary BPA data and the assumptions described in this paper, daily BPA intakes for male participants are statistically significantly higher than for female participants, and there are statistically significant differences in daily BPA intakes according to age groups, with the oldest group having the lowest estimated intakes. Median intake was approximately three orders of magnitude below health-based guidance values of 50 microg/kg-day. PMID- 18414524 TI - Demonstration of chalcogenide glass racetrack microresonators. AB - We have demonstrated what we believe to be the first chalcogenide glass racetrack microresonator using a complementary metal-oxide semiconductor-compatible lift off technique with thermally evaporated As(2)S(3) films. The device simultaneously features a small footprint of 0.012 mm x 0.012 mm, a cavity Q (quality factor) of 10,000, and an extinction ratio of 32 dB. These resonators exhibit a very high sensitivity to refractive index changes with a demonstrated detection capability of Dn(As(2)S(3)=(4.5 x 10(-6)+/-10%) refractive index unit. The resonators were applied to derive a photorefractive response of As(2)S(3) to lambda=550 nm light. The resonator devices are a versatile platform for both sensing and glass material property investigation. PMID- 18414525 TI - Switchable single-longitudinal-mode dual-wavelength erbium-doped fiber ring laser incorporating a semiconductor optical amplifier. AB - We propose and demonstrate a novel single-longitudinal-mode (SLM) dual-wavelength erbium-doped fiber ring laser incorporating a semiconductor optical amplifier. The SOA biased in its low-gain regime greatly reduces the gain competition of the two wavelengths. The stable SLM operation is guaranteed by a passive triple-ring cavity and a fiber Fabry-Perot filter. The dual-wavelength output with a 40 GHz wavelength spacing is switchable in the range of 1533-1565.4 nm. PMID- 18414526 TI - Phase-space quality factor for ultrashort pulsed beams. AB - We derive an expression for the lower bound of the phase-space beam quality factor M(2) of an ultrashort pulse. We show that the condition M(2)>or=1 does not, in general, hold for such pulsed beams. Rather, the minimum value of M(2) depends on the pulse spectrum. We also show that M(2) attains minimum only for pulsed beams whose spot size varies with frequency as omega(-1/2), and that the radial intensity profile of the best-quality pulsed beam can be non-Gaussian. PMID- 18414527 TI - Linear cavity Brillouin fiber laser with improved characteristics. AB - A configuration for linear cavity Brillouin fiber laser (BFL) generation is demonstrated using a standard single-mode fiber, two optical circulators, a 3 dB coupler, and a 95/5 coupler to allow high efficiency. With a Brillouin pump (BP) power of 13 dBm, the laser peak power is 12.3 dB higher than a conventional linear cavity BFL at an upshifted wavelength of 0.086 nm from the BP wavelength. In addition, it is revealed that the BFL peak power can be higher than the transmitted BP peak power when the BP power exceeds the second Brillouin Stokes threshold power. PMID- 18414528 TI - Surface lattice solitons in diffusive nonlinear media. AB - We address the properties of surface solitons supported by optical lattices imprinted in photorefractive media with asymmetrical diffusion nonlinearity. Such solitons exist only in finite gaps of the lattice spectrum. In contrast to latticeless geometries, where surface waves exist only when nonlinearity deflects light toward the material surface, the surface lattice solitons exist in settings where diffusion would cause beam bending against the surface. PMID- 18414529 TI - Simple direct extraction of unknown phase shift and wavefront reconstruction in generalized phase-shifting interferometry: algorithm and experiments. AB - An algorithm to extract the arbitrary unknown phase shift and then reconstruct the complex object wave in generalized phase-shifting interferometry (GPSI) without the iteration process and measurement of object wave intensity is proposed. This method can be used for GPSI of any frame number >or=2. Both computer simulations with smooth and diffusing object surfaces and optical experiments have verified the effectiveness of this method over a wide range of phase shifts with very satisfactory results. PMID- 18414530 TI - Planar infrared binary phase reflectarray. AB - A reflective, binary phase reflectarray is demonstrated in the infrared, at a wavelength of 10.6 microm. The unique aspect of this work, at this frequency band, is that the specific desired phase shift is achieved using an array of subwavelength metallic patches on top of a ground-plane-backed dielectric stand off layer. This is an alternative to the usual method of constructing a reflective Fresnel zone plate by means of a given thickness of dielectric. This initial demonstration of the reflectarray approach at infrared is significant in that there is inherent flexibility to create a range of phase shifts by varying the dimensions of the patches. This will allow for a multilevel phase distribution, or even a continuous variation of phase, across an optical surface with only two-dimensional lithography, avoiding the need for dielectric height variations. PMID- 18414531 TI - Influence of optical heterogeneities on reconstruction of spatial phosphorescence lifetime distributions. AB - We have previously demonstrated that phosphorescence lifetime imaging (PLI) allows for unbiased determination of absolute oxygen levels in homogeneously absorbing/scattering media. We computationally show that tomographic PLI can perform equally well in heterogeneous environments relying only on surface measurements of phosphorescence. PMID- 18414532 TI - Mode matching and insertion loss in ultrabroadband Cr-doped multimode fibers. AB - We investigate the fundamental insertion loss due to mode mismatch in an optical link involving a single-mode fiber-optic (SMF) transmission with the insertion of a segment of a multimode Cr(4+)-doped fiber (MMCDF). With an MMCDF core diameter of approximately 15.5 microm that matches the SMF, we obtained coupling efficiencies of 60.3% to 87.5% in the entire transmission spectral range of 1,300 to 1,600 nm. The high coupling efficiency (or low insertion loss) makes it possible for the employment of the MMCDF as an inline ultrabroadband optical amplifier in an optical link for the entire transmission spectral range. PMID- 18414533 TI - Rapid chemical-vapor sensing using optofluidic ring resonators. AB - We develop rapid chemical-vapor sensors based on optofluidic ring resonators (OFRRs). The OFRR is a glass capillary whose circular wall supports the circulating waveguide modes (WGMs). The OFRR inner surface is coated with a vapor sensitive polymer. The analyte and polymer interaction causes the polymer refractive index to change, which is detected as a WGM spectral shift. Owing to the excellent fluidics, the OFRR exhibits subsecond detection and recovery time with a flow rate of only 1 mL/min, a few orders of magnitude lower than that in the existing optical vapor sensors. The detection limit is estimated to be 5.6 x 10(-6) refractive index units, over ten times better than other ring-resonator vapor sensors. Ethanol and hexane vapors are used as a model system, and chemical differentiation is demonstrated with different polymer coatings. PMID- 18414534 TI - Single-shot acquisition of time-resolved fluorescence spectra using a multiple delay optical fiber bundle. AB - We present a technique for simultaneous acquisition of time-resolved fluorescence spectra using a customized optical fiber bundle with multiple collection fibers of different length. These fibers were aligned at the output plane of a spectrograph corresponding to different wavelengths. Fluorescence signal dispersed by the spectrograph was converted into a pulse train owing to time delays introduced by the length differences between each fiber and subsequently detected by a photomultiplier and digitizer. The performance of the technique was tested with standard fluorescent dyes, and the results are in good agreement with literature values. PMID- 18414535 TI - Direct experimental observation of the single reflection optical Goos-Hanchen shift. AB - We report a precise direct measurement of the Goos-Hanchen shift after one reflection off a dielectric interface coated with periodic metal stripes. The spatial displacement of the shift is determined by image analysis. A maximal absolute shift of 5.18 and 23.39 mum for TE and TM polarized light, respectively, is determined. This technique is simple to implement and can be used for a large range of incident angles. PMID- 18414536 TI - Simultaneous coherent imaging and strain measurement using coupled photorefractive holography and shearography. AB - By coupling photorefractive holography with speckle shearography, it is possible to simultaneously perform both coherent imaging and strain measurement. Use of the photorefractive effect, which is insensitive to incoherently scattered light, is a significant advantage in coherent imaging as described. Experimental results obtained from a centrally loaded steel plate are presented. PMID- 18414537 TI - Spectronanoscopy of photonic wires and supercontinuum generation by parametrically coupled Raman sidebands. AB - Within a narrow spectral region around the wavelength of zero-group-velocity dispersion of a nonlinear-optical waveguide, phase-matched four-wave mixing couples the Stokes Raman sideband of a pump field to its anti-Stokes counterpart. This wave coupling suggests a sensitive probe for linear and nonlinear-optical parameters of the waveguide, enabling the detection of nanoscopic size variations of microchannel waveguides in photonic-crystal fibers, and facilitates the generation of broadband supercontinuum radiation. PMID- 18414538 TI - Generation of polarization-shaped ultraviolet femtosecond pulses. AB - We experimentally demonstrate the generation and characterization of polarization shaped femtosecond laser pulses in the ultraviolet at a central wavelength of 400 nm. Near-infrared laser pulses are first polarization shaped and then frequency doubled in an interferometrically stable setup that employs two perpendicularly oriented nonlinear crystals. A new pulse shaper design involving volume phase holographic gratings reduces losses and hence leads to an increase in pulse energy. PMID- 18414539 TI - Matrix effects on the surface plasmon resonance of dry supported gold nanocrystals. AB - We present a method to characterize surface-chemical properties of gold nanocrystals. Spherical, 60 nm gold nanocrystals were immobilized on quartz substrates by a coupling agent and cleaned in a hydrogen plasma. The nanocrystals were then functionalized with alkanethiol self-assembled monolayers (SAM) of varying chain lengths by adsorption from the gas phase, and localized surface plasmon resonance (LSPR) spectroscopy was performed on the samples. Depending on the alkanethiol chain length, the adsorption of the SAM redshifted the LSPR to different extents, in accordance with Mie theory. SAM thickness differences below 1 nm could be easily resolved. Our results demonstrate that LSPR spectroscopy can be applied to characterize thin organic layers on dry supported gold particles with high sensitivity. PMID- 18414540 TI - Experimental analysis of spectral characteristics of antiresonant guiding photonic crystal fibers. AB - Both numerical and experimental analyses are carried out to investigate the spectral characteristics of antiresonant guiding photonic crystal fibers. The transmission minima were observed at the wavelengths where LP(ml) (m(8)S(7/2)(Gd(3+)), and (3)P(0)/(1)I(6)-->(3)H(6) (Tm(3+)). The energy transfer from Tm(3+) to Gd(3+) plays a crucial role in populating the excited states of Gd(3+). The shortest wavelength of upconverted emission converted from the infrared region was demonstrated here. PMID- 18414557 TI - Approximate optical cloaking in an axisymmetric silicon photonic crystal structure. AB - Axisymmetric photonic crystal structures may be designed to possess interesting optical properties, particularly when the photonic band structure of the material is highly anisotropic. We use finite element calculations to demonstrate an approximate electromagnetic cloaking effect imparted by a structure consisting of concentric silicon photonic crystal layers. The results show that it is possible to bend light around an object by simply using anisotropy. The calculations show that the cloaking mechanism is fundamentally different from Pendry's approach. This design may work as a practical solution for optical cloaking. PMID- 18414558 TI - Ocular aberrations in the peripheral visual field. AB - We modified a commercial Hartmann-Shack aberrometer and used it to measure ocular aberrations twice at each of 38 points across the central 42 degrees horizontal x 32 degrees vertical visual fields of five young emmetropic subjects. Some Zernike aberration coefficients show coefficient field distributions that were similar to the field dependence predicted by Seidel theory (astigmatism, oblique astigmatism, horizontal coma, vertical coma), but defocus did not demonstrate such similarity. PMID- 18414559 TI - Phase recovery in temporal speckle pattern interferometry using the generalized S transform. AB - We propose a novel approach based on the generalized S-transform to retrieve optical phase distributions in temporal speckle pattern interferometry. The performance of the proposed approach is compared with those given by well-known techniques based on the continuous wavelet, the Hilbert transforms, and a smoothed time-frequency distribution by analyzing interferometric data degraded by noise, nonmodulating pixels, and modulation loss. The advantages and limitations of the proposed phase retrieval approach are discussed. PMID- 18414560 TI - All-optical switching in subwavelength metallic grating structure containing nonlinear optical materials. AB - All-optical switching based on a subwavelength metallic grating structure containing nonlinear optical materials has been proposed and numerically investigated. Metal-dielectric composite material is used in the switching for its larger third-order nonlinear susceptibility (approximately 10(-7)esu) and ultrafast response properties. The calculated dependence of the signal light intensity on the pump light intensity shows a bistable behavior, which results in a significant switch effect. It rests on a surface plasmon's enhanced intensity dependent change of the effective dielectric constant of Kerr nonlinear media, corresponding to a transition of the far-field transmission from a low- to high transmission state. The study of this switching structure shows great advantages of smaller size, lower requirement of pump light intensity, and shorter switching time at approximately the picosecond level. PMID- 18414561 TI - Method for acquiring the characteristic parameter of the dual-spiral moire fringes. AB - Based on moire phenomena and the Talbot self-imaging effect, dual-spiral moire fringes could be acquired with two spiral gratings. This kind of special moire fringes could be used to test the collimation of the light beam, which was significant in many correlative applications. The characteristic parameter of the dual-spiral moire fringes reflected the collimation condition directly. A method with two algorithms, which were respectively based on Fourier transform and the phase-shifting algorithm, was proposed and simulated to extract the characteristic parameter. The influence of the random noise on the extraction was also analyzed and discussed. PMID- 18414562 TI - Generation of indistinguishable and pure heralded single photons with tunable bandwidth. AB - We describe a new scheme to fully control the joint spectrum of paired photons generated in spontaneous parametric downconversion. We show the capability of this method to generate frequency-uncorrelated photon pairs that are pure and indistinguishable and whose bandwidth can be readily tuned. Importantly, the scheme we propose can be implemented in any nonlinear crystal and frequency band of interest. PMID- 18414563 TI - Optically induced transition between discrete and gap solitons in a nonconventionally biased photorefractive crystal. AB - We show that optically induced photonic lattices in a nonconventionally biased photorefractive crystal can support the formation of discrete and gap solitons owing to a mechanism that differs from the conventional screening effect. Both the bias direction and the lattice orientation can dramatically influence the nonlinear beam-propagation dynamics. We demonstrate a transition from self focusing to -defocusing and from discrete to gap solitons solely by adjusting the optical-beam orientation. PMID- 18414564 TI - Volume Bragg grating stabilized spectrally narrow Tm fiber laser. AB - A Tm-doped large mode area (LMA) silica fiber laser has been locked to a stable wavelength of 2,053.9 nm using a volume Bragg grating (VBG). The measured spectral width of the laser output was <300 pm, limited by the spectrometer resolution. Although this laser had modest output (approximately 5W) and slope efficiency (41%), this new approach to spectrally limiting the output of LMA fiber lasers is inherently extendable to kilowatt powers, opening up several applications including high-power pulsed fiber lasers and spectral beam combining. Performance characteristics of the laser compared to one using a dielectric mirror as a high reflector are described, and the results imply low VBG losses. PMID- 18414565 TI - Integration of grating couplers with a compact photonic crystal demultiplexer on an InP membrane. AB - We demonstrate the integration of a 30% efficient grating coupler with a compact photonic crystal wavelength demultiplexer (DeMUX). The DeMUX has seven output channels that are spaced 10 nm apart and is aimed at coarse WDM applications. The integrated devices are realized on a high-index-contrast InP membrane using a simple benzocyclobutene wafer bonding technique. Cross talks of -10 to -12 dB for four channels 20 nm apart are obtained without optimization. PMID- 18414566 TI - Measurement of the second hyperpolarizability of linear conjugated polymer based on attenuated-total-reflection technique. AB - Based on the attenuated-total-reflection technique, a new method has been proposed to determine the second hyperpolarizability gamma( omega(4);omega(3),omega(2),omega(1)) of the linear conjugated polymer in the off resonant region by means of quadratic electro-optic effect. An important feature of this method is the absence of a high-power pulse laser, resulting in more convenience and cost effectivity than other techniques. PMID- 18414567 TI - 10 GHz, 2.4 ps pulse generation using a single-stage dual-drive Mach-Zehnder modulator. AB - This Letter reports on picosecond pulse generation at a repetition of 10 GHz by using a single-stage electro-optic LiNbO(3) Mach-Zehnder modulator, stressing the simplicity of its setup. It is analytically and experimentally proved that dual arm modulation with in-phase sinusoidal signals having slightly different amplitudes generated a highly coherent optical comb with a great spectral flatness and a parabolic phase relationship in its spectrum. The generated comb was Fourier synthesized and shaped into an ultrashort pulse train with an optical bandpass filter and a dispersive fiber. The pulse source was highly stable, exhibiting an ultralow timing jitter of less than 130 fs. PMID- 18414584 TI - Nuclear Factor-kappa B as a Resistance Factor to Platinum-Based Antineoplasic Drugs. AB - Platinum drugs continue to be major chemotherapy drugs for cancer treatment. Nevertheless, acquired or intrinsic resistance to these compounds is common in human tumors. One important mechanism for this resistance is the avoidance of cells entering the apoptotic pathway. Nuclear factor-kappa B (NF-kappa B, NF kappaB) is a pleiotropic transcription factor key in determining the death threshold of human cells. This factor is important in the final response of cells to platinum drugs, as exemplified by in vitro and in vivo models showing that inhibition of NF-kappaB sensitizes cancer cells to the effects of these drugs. New approaches focusing on the inhibition of NF-kappaB could help to minimize or even eliminate intrinsic or acquired resistance to platinum drugs. PMID- 18414585 TI - A scaffold analysis tool using mate-pair information in genome sequencing. AB - We have developed a Windows-based program, ConPath, as a scaffold analyzer. ConPath constructs scaffolds by ordering and orienting separate sequence contigs by exploiting the mate-pair information between contig-pairs. Our algorithm builds directed graphs from link information and traverses them to find the longest acyclic graphs. Using end read pairs of fixed-sized mate-pair libraries, ConPath determines relative orientations of all contigs, estimates the gap size of each adjacent contig pair, and reports wrong assembly information by validating orientations and gap sizes. We have utilized ConPath in more than 10 microbial genome projects, including Mannheimia succiniciproducens and Vibro vulnificus, where we verified contig assembly and identified several erroneous contigs using the four types of error defined in ConPath. Also, ConPath supports some convenient features and viewers that permit investigation of each contig in detail; these include contig viewer, scaffold viewer, edge information list, mate pair list, and the printing of complex scaffold structures. PMID- 18414586 TI - Synthesis, Spectroscopic, and Anticancerous Properties of Mixed Ligand Palladium(II) and Silver(I) Complexes with 4,6-Diamino-5-hydroxy-2 mercaptopyrimidine and 2,2'-Bipyridyl. AB - Synthesis of two new water-soluble mixed ligand [Pd(bpy)(dahmp)]Cl and [Ag(bpy)(Hdahmp)]NO(3) complexes (dahmp and Hdahmp are the deprotonated monoanion and the protonated neutral 4,6-diamino-5-hydroxy-2-mercaptopyrimidine, resp.) is reported. The composition of the reported complexes was discussed on the bases of IR, (1)H NMR, and mass spectra, as well as conductivity and thermal measurements. The reported complexes display a significant anticancer activity against Ehrlich ascites tumor cells (EACs). The higher activity of these complexes with their higher conductivity values corresponds to their complete ionization in aqueous solution. PMID- 18414589 TI - Brucella abortus strain RB51 vaccine: immune response after calfhood vaccination and field investigation in Italian cattle population. AB - Immune response to Brucella abortus strain RB51 vaccine was measured in cattle vaccinated at calfhood. After an increase at day 6 post-vaccination (pv), the antibody level recorded in the 10 vaccinated animals remained constant for two months, and then progressively decreased. All vaccinated animals remained negative from day 162 pv to the end of the study (day 300 pv). Only at days 13 and 14 pv the RB51-CFT showed 100% sensitivity (credibility interval (CI) 76.2% 100%). The results indicate that the possibility to use RB51-CFT for the identification of cattle vaccinated at calfhood with RB51 is limited in time. A field investigation was carried out on 26,975 sera collected on regional basis from the Italian cattle population. The study outcomes indicate that in case of RB51-CFT positive results observed in officially Brucellosis-free (OBF) areas and, in any case, when an illegal use of RB51 vaccine is suspected, the use of the RB51-CFT alone is not sufficient to identify all the vaccinated animals. The design of a more sophisticated diagnostic protocol including an epidemiological investigation, the use of RB51-CFT, and the use of the skin test with RB51 as antigen is deemed more appropriate for the identification of RB51 vaccinated animals. PMID- 18414587 TI - Novel Anticancer Platinum(IV) Complexes with Adamantylamine: Their Efficiency and Innovative Chemotherapy Strategies Modifying Lipid Metabolism. AB - The impressive impact of cisplatin on cancer on one side and severe side effects, as well as the development of drug resistance during treatment on the other side, were the factors motivating scientists to design and synthesize new more potent analogues lacking disadvantages of cisplatin. Platinum(IV) complexes represent one of the perspective groups of platinum-based drugs. In this review, we summarize recent findings on both in vitro and in vivo effects of platinum(IV) complexes with adamantylamine. Based on a literary overview of the mechanisms of activity of platinum-based cytostatics, we discuss opportunities for modulating the effects of novel platinum complexes through interactions with apoptotic signaling pathways and with cellular lipids, including modulations of the mitochondrial cell death pathway, oxidative stress, signaling of death ligands, lipid metabolism/signaling, or intercellular communication. These approaches might significantly enhance the efficacy of both novel and established platinum based cytostatics. PMID- 18414590 TI - Primary malignant tumours of bone following previous malignancy. AB - Destructive bone lesions occurring in patients who have previously had a malignancy are generally assumed to be a metastasis from that malignancy. We reviewed 60 patients with a previous history of malignancy, who presented with a solitary bone lesion that was subsequently found to be a new and different primary sarcoma of bone. These second malignancies occurred in three distinct groups of patients: (1) patients with original tumours well known to be associated with second malignancies (5%); (2) patients whose second malignancies were likely to be due to the previous treatment of their primary malignancy (40%); (3) patients in whom there was no clearly defined association between malignancies (55%). The purpose of this study is to emphasise the necessity for caution in assuming the diagnosis of a metastasis when a solitary bone lesion is identified following a prior malignancy. Inappropriate biopsy and treatment of primary bone sarcomas compromises limb salvage surgery and can affect patient mortality. PMID- 18414588 TI - Mouse mammary tumor virus promoter-containing retroviral promoter conversion vectors for gene-directed enzyme prodrug therapy are functional in vitro and in vivo. AB - Gene directed-enzyme prodrug therapy (GDEPT) is an approach for sensitization of tumor cells to an enzymatically activated, otherwise nontoxic, prodrug. Cytochrome P450 2B1 (CYP2B1) metabolizes the prodrugs cyclophosphamide (CPA) and ifosfamide (IFA) to produce the cytotoxic substances phosphoramide mustard and isophosphoramide mustard as well as the byproduct acrolein. We have constructed a retroviral promoter conversion (ProCon) vector for breast cancer GDEPT. The vector allows expression of CYP2B1 from the mouse mammary tumor virus (MMTV) promoter known to be active in the mammary glands of transgenic animals. It is anticipated to be used for the generation of encapsulated viral vector producing cells which, when placed inside or close to a tumor, will act as suppliers of the therapeutic CYP2B1 protein as well as of the therapeutic vector itself. The generated vector was effectively packaged by virus producing cells and allowed the production of high levels of enzymatically active CYP2B1 in infected cells which sensitized them to killing upon treatment with both IFA and CPA. Determination of the respective IC(50) values demonstrated that the effective IFA dose was reduced by sixteen folds. Infection efficiencies in vivo were determined using a reporter gene-bearing vector in a mammary cancer cell-derived xenograft tumor mouse model. PMID- 18414591 TI - Wound healing in a patient with psoriasis vulgaris and femur megaprosthesis implantation. AB - Extraskeletal mesenchymal chondrosarcoma is extremely rare and, in combination with psoriasis, it has never been described before. We report a case of wide resection of an extraskeletal chondrosarcoma of the thigh and reconstruction with a femoral megaprosthesis in a patient with psoriasis vulgaris. Special emphasis has been laid to postoperative wound healing in psoriatic skin which did not show any problems. PMID- 18414592 TI - Spin valve sensors for ultrasensitive detection of superparamagnetic nanoparticles for biological applications. AB - We present giant magnetoresistance (GMR) spin valve sensors designed for detection of superparamagnetic nanoparticles as potential biomolecular labels in magnetic biodetection technology. We discuss the sensor design and experimentally demonstrate that as few as approximately 23 monodisperse 16-nm superparamagnetic Fe(3)O(4) nanoparticles can be detected by submicron spin valve sensors at room temperature without resorting to lock-in detection. A patterned self-assembly method of nanoparticles, based on a polymer-mediated process and fine lithography, is developed for the detection. It is found that sensor signal increases linearly with the number of nanoparticles. PMID- 18414593 TI - From single motor unit activity to multiple grip forces: mini-review of multi digit grasping. AB - SYNOPSIS: This paper is a mini review of kinetic and kinematic evidence on the control of the hand with emphasis on grasping. It is not meant to be an exhaustive review, rather it summarizes current research examining the mechanisms through which specific patterns of coordination are elicited and observed during reach to grasp movements and static grasping. These coordination patterns include the spatial and temporal covariation of the rotation at multiple joints during reach to grasp movements. A basic coordination between grip forces produced by multiple digits also occurs during whole hand grasping such that normal forces tend to be produced in a synchronous fashion across pairs of digits. Finally, we address current research that suggests that motor unit synchrony across hand muscles and muscle compartments might be one of the neural mechanisms underlying the control of grasping. PMID- 18414594 TI - Skating down a steeper slope: fear influences the perception of geographical slant. AB - Previous studies have shown that conscious awareness of hill slant is overestimated, but visually guided actions directed at hills are relatively accurate. Also, steep hills are consciously estimated to be steeper from the top than the bottom, possibly because they are dangerous to descend. In the present study, participants stood at the top of a hill either on a skateboard or a wooden box of the same height. They gave three estimates of the slant: a verbal report, a visually matched estimate, and a visually guided action. Fear of descending the hill was also assessed. Those participants who were scared (by the skateboard) consciously judged the hill to be steeper than unafraid participants. However, the visually guided action measure was accurate across conditions. These results suggest that explicit awareness of slant is influenced by the fear associated with a potentially dangerous action that could be performed on the hill. PMID- 18414595 TI - Siblings' Differential Treatment in Mexican American Families. AB - We investigated the patterns and correlates of parents' differential treatment of adolescent siblings in 246 two-parent Mexican American families. In home interviews, siblings rated 7 domains of differential treatment (e.g., privileges, chores, warmth) as well as their adjustment and perceptions of parental acceptance and fairness, and both parents and adolescents reported on cultural dynamics. More gender-typed patterns of differential treatment were evident when parents were more oriented to Mexican than Anglo culture. The links between differential treatment and youth reports of adjustment, parental acceptance, and parental fairness were moderated by adolescents' familism values, particularly for older siblings: Differential treatment was linked more strongly to adjustment and parent-youth relationship problems when youth reported lower levels of familism. PMID- 18414596 TI - Mexican American Fathers' Occupational Conditions: Links to Family Members' Psychological Adjustment. AB - To examine the implications of fathers' occupational conditions (i.e., income, work hours, shift work, pressure, workplace racism, and underemployment) for family members' psychological adjustment, home interviews were conducted with fathers, mothers, and two adolescent offspring in each of 218 Mexican American families. Results underscored the importance of acculturation as a moderator. Fathers' income was negatively associated with depressive symptoms in highly acculturated families but not in less acculturated families. In contrast, fathers' reports of workplace racism were positively associated with depressive symptoms in less acculturated families but not in more acculturated family contexts. These findings were consistent across all 4 family members, suggesting that the "long arm" of the jobs held by Mexican American fathers extends to mothers and adolescent offspring. PMID- 18414598 TI - Lunatic fringe causes expansion and increased neurogenesis of trunk neural tube and neural crest populations. AB - Both neurons and glia of the PNS are derived from the neural crest. In this study, we have examined the potential function of lunatic fringe in neural tube and trunk neural crest development by gain-of-function analysis during early stages of nervous system formation. Normally lunatic fringe is expressed in three broad bands within the neural tube, and is most prominent in the dorsal neural tube containing neural crest precursors. Using retrovirally-mediated gene transfer, we find that excess lunatic fringe in the neural tube increases the numbers of neural crest cells in the migratory stream via an apparent increase in cell proliferation. In addition, lunatic fringe augments the numbers of neurons and upregulates Delta-1 expression. The results indicate that, by modulating Notch/Delta signaling, lunatic fringe not only increases cell division of neural crest precursors, but also increases the numbers of neurons in the trunk neural crest. PMID- 18414600 TI - Aging and Language Production. AB - Experimental research and older adults' reports of their own experience suggest that the ability to produce the spoken forms of familiar words declines with aging. Older adults experience more word-finding failures, such as tip-of-the tongue states, than young adults do, and this and other speech production failures appear to stem from difficulties in retrieving the sounds of words. Recent evidence has identified a parallel age-related decline in retrieving the spelling of familiar words. Models of cognitive aging must explain why these aspects of language production decline with aging whereas semantic processes are well maintained. We describe a model wherein aging weakens connections among linguistic representations, thereby reducing the transmission of excitation from one representation to another. The structure of the representational systems for word phonology and orthography makes them vulnerable to transmission deficits, impairing retrieval. PMID- 18414599 TI - Amygdala contribution to selective dimensions of emotion. AB - The amygdala has been implicated in emotional processes, although the precise nature of the emotional deficits following amygdala lesions remains to be fully elucidated. Cognitive disturbances in the perception, recognition or memory of emotional stimuli have been suggested by some, whereas others have proposed changes in emotional arousal. To address this issue, measures of emotional arousal and valence (positivity and negativity) to a graded series of emotional pictures were obtained from patients with lesions of the amygdala and from a clinical contrast group with lesions that spared this structure. Relative to the contrast group, patients with damage to the amygdala evidenced a complete lack of an arousal gradient across negative stimuli, although they displayed a typical arousal gradient to positive stimuli. These results were not attributable to the inability of amygdala patients to process the hostile or hospitable nature of the stimuli, as the amygdala group accurately recognized and categorized both positive and negative features of the stimuli. The relative lack of emotional arousal to negative stimuli may account for many of the clinical features of amygdala lesions. PMID- 18414601 TI - A new day for global environmental health. PMID- 18414602 TI - The Cuyahoga is still burning. PMID- 18414604 TI - Aggregation and toxicology of titanium dioxide nanoparticles. PMID- 18414606 TI - DDT and breast cancer. PMID- 18414607 TI - Comments on "the sweet scent on baby's breath?". PMID- 18414608 TI - The interaction of agricultural pesticides and marginal iodine nutrition status as a cause of autism spectrum disorders. PMID- 18414609 TI - Chemical exposures: an eye to the sea. PMID- 18414615 TI - Benefits of sunlight: a bright spot for human health. PMID- 18414616 TI - Drought in the Southeast: lessons for water management. PMID- 18414617 TI - Arsenic and erectile dysfunction: drinking contaminated well water increases risk. PMID- 18414618 TI - Better than eating worms? Children's dietary exposure to OP pesticides. PMID- 18414619 TI - Bleached, but not by the sun: sunscreen linked to coral damage. PMID- 18414620 TI - Cooking with wood may fuel low birth weight: kitchen smoke puts babies at risk. PMID- 18414621 TI - NIEHS extramural global environmental health portfolio: opportunities for collaboration. AB - BACKGROUND: Global environmental health has emerged as a critical topic for environmental health researchers and practitioners. Estimates of the environmental contribution of total worldwide disease burden range from 25 to 33%. OBJECTIVE: We reviewed grants funded by the National Institute of Environmental Health Sciences (NIEHS) during 2005-2007 to evaluate the costs and scientific composition of the global environmental health portfolio, with the ultimate aim of strengthening global environmental health research partnerships. METHODS/RESULTS: We examined NIEHS grant research databases to identify the global environmental health portfolio. In the past 3 fiscal years (2005-2007), the NIEHS funded 57 scientific research projects in 37 countries, at an estimated cost of $30 million. Metals such as arsenic, methylmercury, and lead are the most frequently studied toxic agents, but a wide range of stressors, routes of exposure, and agents are addressed in the portfolio. CONCLUSIONS: The portfolio analysis indicates that there is a firm foundation of research activities upon which additional global environmental health partnerships could be encouraged. Current data structures could be strengthened to support more automated analysis of grantee information. PMID- 18414623 TI - Transformation of human urothelial cells (UROtsa) by as and cd induces the expression of keratin 6a. AB - BACKGROUND: Cadmium and arsenite can directly and malignantly transform the UROtsa cell line. The tumor heterotransplants produced from these transformed cells have histologic features consistent with human bladder cancer. Previous microarray analysis of total RNA from the parental and transformed cells suggested that keratin 6a was overexpressed as a result of cell transformation. OBJECTIVES: Our goals were to verify overexpression of keratin 6a in Cd(2+)- and As(3+)-transformed UROtsa cells, the corresponding tumor heterotransplants, and human bladder cancer biopsy specimens and to assess what factors may be involved in keratin 6a overexpression. METHODS: Expression was assessed with real-time polymerase chain reaction, Western blot analysis, and immunohistochemistry. We used the effect of addition and deletion of potential growth factors in the cell culture growth medium to assess possible pathways used in keratin 6a overexpression. RESULTS: Cd(2+)- and As(3+)-transformed cells grown in serum containing growth medium, as well as the derived tumor heterotransplants, overexpressed keratin 6a mRNA and protein compared with UROtsa cells grown in serum-containing growth medium. Immunostaining of keratin 6a in tumor heterotransplants showed focal staining of the tumor cells that was localized to the cytoplasm. Focal immunostaining of keratin 6a was also found in some but not all archival patient specimens of high-grade bladder cancer, confirming translation of the results to human bladder cancer. Studies on growth factor deletion and addition indicated that the level of keratin 6a expression was regulated by the presence of both insulin and epidermal growth factor (EGF). In contrast, growth factors had no effect on the elevated levels of keratin 6a expression found in transformed UROtsa cells. CONCLUSIONS: Our present studies suggest that keratin 6a expression may be a biomarker for malignant urothelial cells that possess an activated EGF and or insulin growth factor pathway. PMID- 18414622 TI - Phytoestrogens and breast cancer prevention: possible mechanisms of action. AB - OBJECTIVE: Phytoestrogens display an array of pharmacologic properties, and in recent years investigation of their potential as anticancer agents has increased dramatically. In this article we review the published literature related to phytoestrogens and breast cancer as well as suggest the possible mechanisms that may underlie the relationship between phytoestrogens and breast cancer. DATA SOURCES: Electronic searches on phytoestrogens and breast cancer were performed on MEDLINE and EMBASE in June 2007. No date restriction was placed on the electronic search. DATA EXTRACTION: We focused on experimental data from published studies that examined the characteristics of phytoestrogens using in vivo or in vitro models. We also include human intervention studies in this review. DATA SYNTHESIS: We evaluated evidence regarding the possible mechanisms of phytoestrogen action. Discussions of these mechanisms were organized into those activities related to the estrogen receptor, cell growth and proliferation, tumor development, signaling pathways, and estrogen-metabolizing enzymes. CONCLUSIONS: We suggest that despite numerous investigations, the mechanisms of phytoestrogen action in breast cancer have yet to be elucidated. It remains uncertain whether these plant compounds are chemoprotective or whether they may produce adverse outcomes related to breast carcinogenesis. PMID- 18414624 TI - Sunscreens cause coral bleaching by promoting viral infections. AB - BACKGROUND: Coral bleaching (i.e., the release of coral symbiotic zooxanthellae) has negative impacts on biodiversity and functioning of reef ecosystems and their production of goods and services. This increasing world-wide phenomenon is associated with temperature anomalies, high irradiance, pollution, and bacterial diseases. Recently, it has been demonstrated that personal care products, including sunscreens, have an impact on aquatic organisms similar to that of other contaminants. OBJECTIVES: Our goal was to evaluate the potential impact of sunscreen ingredients on hard corals and their symbiotic algae. METHODS: In situ and laboratory experiments were conducted in several tropical regions (the Atlantic, Indian, and Pacific Oceans, and the Red Sea) by supplementing coral branches with aliquots of sunscreens and common ultraviolet filters contained in sunscreen formula. Zooxanthellae were checked for viral infection by epifluorescence and transmission electron microscopy analyses. RESULTS: Sunscreens cause the rapid and complete bleaching of hard corals, even at extremely low concentrations. The effect of sunscreens is due to organic ultraviolet filters, which are able to induce the lytic viral cycle in symbiotic zooxanthellae with latent infections. CONCLUSIONS: We conclude that sunscreens, by promoting viral infection, potentially play an important role in coral bleaching in areas prone to high levels of recreational use by humans. PMID- 18414625 TI - Contamination of potable water distribution systems by multiantimicrobial resistant enterohemorrhagic Escherichia coli. AB - BACKGROUND: The contamination of processed or unprocessed drinking water by fecal coliform bacteria has been reported worldwide. Despite a high incidence of waterborne diseases, entero-hemorrhagic Escherichia coli (EHEC) is an underacknowledged pathogen of concern to public health in India. Although the presence of EHEC is recorded in surface water resources of India, drinking water sources are yet to be investigated. OBJECTIVES: The goal of this study was to analyze potable water samples for the presence of virulence determinants of EHEC and to determine the sensitivity of the virulence determinants to antimicrobials. METHODS: We enumerated coliform bacteria in potable water samples collected from six locations in Lucknow, a major city in northern India, using the most probable number method. E. coli (n = 81), randomly isolated by membrane-filtration technique from four sites, were identified by biochemical characterization. E. coli were not detected in samples from two other sites. We screened 15 randomly selected isolates from each site for virulence determinants of EHEC using polymerase chain reaction (PCR). The isolates positive for virulence determinants (n = 18) were screened for sensitivity to 15 antimicrobials by the disk diffusion method. RESULTS: Both stx1 and stx2 genes were present in 33.3% of isolates, whereas others possessed either stx1 (11.1%) or stx2 (55.6%). eaeA, hlyA, and chuA genes were present in 100, 23.3, and 16.7% of isolates, respectively. Resistance to multiple antimicrobials was observed in potential EHEC. CONCLUSIONS: The occurrence of multiantimicrobial-resistant EHEC in potable water is an important health concern because of the risk of waterborne outbreaks. PMID- 18414626 TI - The impact of secondary particles on the association between ambient ozone and mortality. AB - BACKGROUND: Although several previous studies have found a positive association between ambient ozone and mortality, the observed effect may be confounded by other secondary pollutants that are produced concurrently with ozone. OBJECTIVES: We addressed the question of whether the ozone-mortality relationship is entirely a reflection of the adverse effect of ozone, or whether it is, at least in part, an effect of other secondary pollutants. METHODS: Separate time-series models were fit to 3-6 years of data between 2000 and 2005 from 18 U.S. communities. The association between nonaccidental mortality was examined with ozone alone and with ozone after adjustment for fine particle mass, sulfate, organic carbon, or nitrate concentrations. The effect estimates from each of these models were pooled using a random-effects meta-analysis to obtain an across-community average. RESULTS: We found a 0.89% [95% confidence interval (CI), 0.45-1.33%] increase in nonaccidental mortality with a 10-ppb increase in same-day 24-hr summertime ozone across the 18 communities. After adjustment for PM(2.5) (particulate matter with aerodynamic diameter or= 50 years of age through health examinations conducted in three hospitals in Taiwan. We used a questionnaire (International Index of Erectile Function-5) to measure the level of erectile function. Sex hormones, including total testosterone and sex hormone-binding globulin, were determined by radioimmunoassay. We used another standardized questionnaire to collect background and behavioral information (e.g., cigarette smoking; alcohol, tea, or coffee drinking; and physical activity). RESULTS: The prevalence of ED was greater in the arsenic-endemic area (83.3%) than in the non-arsenic-endemic area (66.7%). Subjects with arsenic exposure > 50 ppb had a significantly higher risk of developing ED than those with exposure or= 60% of the total county acreage devoted to farming. Risk for different cancers varied by type of crop. CONCLUSIONS: Although interpretation is limited by the ecologic design, in this study we were able to evaluate rarer childhood cancers across a diverse agricultural topography. The findings of this exploratory study support a continued interest in the possible impact of long-term, low-level pesticide exposure in communities located in agriculturally intense areas. PMID- 18414644 TI - Impaired reproductive development in sons of women occupationally exposed to pesticides during pregnancy. AB - OBJECTIVES: The aim of this prospective study was to investigate whether occupational pesticide exposure during pregnancy causes adverse effects on the reproductive development in the male infants. DESIGN AND MEASUREMENTS: Pregnant women employed in greenhouses in Denmark were consecutively recruited, and 113 mother-son pairs were included. The mothers were categorized as occupationally exposed (91 sons) or unexposed (22 sons) to pesticides during pregnancy. Testicular position and volume, penile length, and position of urethral opening were determined at 3 months of age using standardized techniques. Concentrations of reproductive hormones in serum from the boys were analyzed. RESULTS: The prevalence of cryptorchidism at 3 months of age was 6.2% [95% confidence interval (CI), 3.0-12.4]. This prevalence was considerably higher than among Danish boys born in the Copenhagen area (1.9%; 95% CI, 1.2-3.0) examined by the same procedure. Boys of pesticide-exposed mothers showed decreased penile length, testicular volume, serum concentrations of testosterone, and inhibin B. Serum concentrations of sex hormone-binding globulin, follicle-stimulating hormone, and the luteinizing hormone:testosterone ratio were increased compared with boys of nonexposed mothers. For individual parameters, only the decreased penile length was statistically significant (p = 0.04). However, all observed effects were in the anticipated direction, and a joint multivariate test showed that this finding had a p-value of 0.012. CONCLUSIONS: Our findings suggest an adverse effect of maternal occupational pesticide exposure on reproductive development in the sons despite current greenhouse safeguards and special measures to protect pregnant women. PMID- 18414645 TI - A unique quinolineboronic acid-based supramolecular structure that relies on double intermolecular B-N bonds for self-assembly in solid state and in solution. AB - The boronic acid functional group plays very important roles in sugar recognition, catalysis, organic synthesis, and supramolecular assembly. Therefore, understanding the unique properties of this functional group is very important. 8-Quinolineboronic acid (8-QBA) is found to be capable of self assembling in solid state through a unique intermolecular B-N bond mechanism reinforced by intermolecular boronic anhydride formation, pi-pi stacking, and hydrogen bond formation. NMR NOE and diffusion studies indicate that intermolecular B-N interaction also exists in solution with 8-QBA. In contrast, a positional isomer of 8-QBA, 5-quinolineboronic acid (5-QBA) showed very different behaviors in crystal packing and in solution and therefore different supramolecular network. Understanding the structural features of this unique 8 QBA assembly could be very helpful for the future design of new sugar sensors, molecular catalysts, and supramolecular assemblies. PMID- 18414647 TI - Effects of deworming on malnourished preschool children in India: an open labelled, cluster-randomized trial. AB - BACKGROUND: More than a third of the world's children are infected with intestinal nematodes. Current control approaches emphasise treatment of school age children, and there is a lack of information on the effects of deworming preschool children. METHODOLOGY: We studied the effects on the heights and weights of 3,935 children, initially 1 to 5 years of age, of five rounds of anthelmintic treatment (400 mg albendazole) administered every 6 months over 2 years. The children lived in 50 areas, each defined by precise government boundaries as urban slums, in Lucknow, North India. All children were offered vitamin A every 6 months, and children in 25 randomly assigned slum areas also received 6-monthly albendazole. Treatments were delivered by the State Integrated Child Development Scheme (ICDS), and height and weight were monitored at baseline and every 6 months for 24 months (trial registration number NCT00396500). p Value calculations are based only on the 50 area-specific mean values, as randomization was by area. FINDINGS: The ICDS infrastructure proved able to deliver the interventions. 95% (3,712/3,912) of those alive at the end of the study had received all five interventions and had been measured during all four follow-up surveys, and 99% (3,855/3,912) were measured at the last of these surveys. At this final follow up, the albendazole-treated arm exhibited a similar height gain but a 35 (SE 5) % greater weight gain, equivalent to an extra 1 (SE 0.15) kg over 2 years (99% CI 0.6-1.4 kg, p = 10(-11)). CONCLUSIONS: In such urban slums in the 1990s, five 6-monthly rounds of single dose anthelmintic treatment of malnourished, poor children initially aged 1-5 years results in substantial weight gain. The ICDS system could provide a sustainable, inexpensive approach to the delivery of anthelmintics or micronutrient supplements to such populations. As, however, we do not know the control parasite burden, these results are difficult to generalize. TRIAL REGISTRATION: ClinicalTrials.gov NCT00396500. PMID- 18414646 TI - Interactions between natural populations of human and rodent schistosomes in the Lake Victoria region of Kenya: a molecular epidemiological approach. AB - BACKGROUND: Schistosoma mansoni exists in a complex environmental milieu that may select for significant evolutionary changes in this species. In Kenya, the sympatric distribution of S. mansoni with S. rodhaini potentially influences the epidemiology, ecology, and evolutionary biology of both species, because they infect the same species of snail and mammalian hosts and are capable of hybridization. METHODOLOGY/PRINCIPAL FINDINGS: Over a 2-year period, using a molecular epidemiological approach, we examined spatial and temporal distributions, and the overlap of these schistosomes within snails, in natural settings in Kenya. Both species had spatially and temporally patchy distributions, although S. mansoni was eight times more common than S. rodhaini. Both species were overdispersed within snails, and most snails (85.2% for S. mansoni and 91.7% for S. rodhaini) only harbored one schistosome genotype. Over time, half of snails infected with multiple genotypes showed a replacement pattern in which an initially dominant genotype was less represented in later replicates. The other half showed a consistent pattern over time; however, the ratio of each genotype was skewed. Profiles of circadian emergence of cercariae revealed that S. rodhaini emerges throughout the 24-hour cycle, with peak emergence before sunrise and sometimes immediately after sunset, which differs from previous reports of a single nocturnal peak immediately after sunset. Peak emergence for S. mansoni cercariae occurred as light became most intense and overlapped temporally with S. rodhaini. Comparison of schistosome communities within snails against a null model indicated that the community was structured and that coinfections were more common than expected by chance. In mixed infections, cercarial emergence over 24 hours remained similar to single species infections, again with S. rodhaini and S. mansoni cercarial emergence profiles overlapping substantially. CONCLUSIONS/SIGNIFICANCE: The data from this study indicate a lack of obvious spatial or temporal isolating mechanisms to prevent hybridization, raising the intriguing question of how the two species retain their separate identities. PMID- 18414648 TI - Immunity to distinct sand fly salivary proteins primes the anti-Leishmania immune response towards protection or exacerbation of disease. AB - BACKGROUND: Leishmania parasites are transmitted in the presence of sand fly saliva. Together with the parasite, the sand fly injects biologically active salivary components that favorably change the environment at the feeding site. Exposure to bites or to salivary proteins results in immunity specific to these components. Mice immunized with Phlebotomus papatasi salivary gland homogenate (SGH) or pre-exposed to uninfected bites were protected against Leishmania major infection delivered by needle inoculation with SGH or by infected sand fly bites. Immunization with individual salivary proteins of two sand fly species protected mice from L. major infection. Here, we analyze the immune response to distinct salivary proteins from P. papatasi that produced contrasting outcomes of L. major infection. METHODOLOGY/PRINCIPAL FINDINGS: DNA immunization with distinct DTH inducing salivary proteins from P. papatasi modulates L. major infection. PpSP15 immunized mice (PpSP15-mice) show lasting protection while PpSP44-immunized mice (PpSP44-mice) aggravate the infection, suggesting that immunization with these distinct molecules alters the course of anti-Leishmania immunity. Two weeks post infection, 31.5% of CD4(+) T cells produced IFN-gamma in PpSP15-mice compared to 7.1% in PpSP44-mice. Moreover, IL-4-producing cells were 3-fold higher in PpSP44 mice. At an earlier time point of two hours after challenge with SGH and L. major, the expression profile of PpSP15-mice showed over 3-fold higher IFN-gamma and IL-12-Rbeta2 and 20-fold lower IL-4 expression relative to PpSP44-mice, suggesting that salivary proteins differentially prime anti-Leishmania immunity. This immune response is inducible by sand fly bites where PpSP15-mice showed a 3 fold higher IFN-gamma and a 5-fold lower IL-4 expression compared with PpSP44 mice. CONCLUSIONS/SIGNIFICANCE: Immunization with two salivary proteins from P. papatasi, PpSP15 and PpSP44, produced distinct immune profiles that correlated with resistance or susceptibility to Leishmania infection. The demonstration for the first time that immunity to a defined salivary protein (PpSP44) results in disease enhancement stresses the importance of the proper selection of vector based vaccine candidates. PMID- 18414649 TI - Lower expression of TLR2 and SOCS-3 is associated with Schistosoma haematobium infection and with lower risk for allergic reactivity in children living in a rural area in Ghana. AB - BACKGROUND: Helminth infections are prevalent in rural areas of developing countries and have in some studies been negatively associated with allergic disorders and atopy. In this context little is known of the molecular mechanisms of modulation involved. We have characterized the innate immune responses, at the molecular level, in children according to their helminth infection status and their atopic reactivity to allergens. METHODOLOGY/PRINCIPAL FINDINGS: The mRNA expression of several genes of the innate immune system that have been associated with microbial exposure and allergy was examined in 120 school children in a rural area in Ghana. Helminth infections were common and atopy rare in the study area. The analysis of gene expression in ex vivo whole blood samples reflected the levels of corresponding proteins. Using this approach in a population of school children in whom the presence of Schistosoma haematobium infection was associated with protection from atopic reactivity, we found that the level of TLR2 and SOCS-3, genes associated with atopy in the children, were significantly downregulated by presence of S. haematobium infection. CONCLUSIONS: S. haematobium infections modulate the expression of genes of the innate immune system (TLR2 and SOCS-3); these are genes that are associated with increased allergic inflammatory processes, providing a molecular link between the negative association of this infection and atopy in rural children in Ghana. PMID- 18414650 TI - TPO, but not soluble-IL-6 receptor, levels increase after anagrelide treatment of thrombocythemia in chronic myeloproliferative disorders. AB - Anagrelide is often used in the treatment of thrombocythemia in myeloproliferative disease (MPD), but information concerning effects of treatment on cytokines involved in regulation of blood platelet levels is limited. Here, we investigated serum levels of thrombopoietin (TPO) and soluble IL-6 receptor (sIL 6R) in relation to response to treatment with and plasma concentrations of anagrelide. Samples from 45 patients with thrombocythemia due to MPD (ET=31, PV=14), being treated with anagrelide for 6 months, were analyzed for TPO, sIL-6R and anagrelide levels. The mean baseline platelet count was 983x10(9)/L. A reduction of platelets to <600 in asymptomatic or <400 x 10(9)/L in symptomatic patients was defined as a complete remission (CR), a reduction with >50% of baseline as partial remission, and <50% reduction as failure. At 6 months, 35 patients were in CR, 1 had a partial remission and 9 were treatment failures. For all patients, there was an increase in TPO of 44% from baseline; this change was more pronounced for patients with partial remission and failure. sIL-6R levels did not change significantly. There was no correlation between levels of anagrelide and cytokine levels at 6 months, and changes of cytokine levels did not relate to changes of platelet counts. Thus, a pronounced increase of TPO levels after 6 months of anagrelide treatment indicated that this treatment affected a major regulatory mechanism for megakaryocyte and platelet formation in MPD. PMID- 18414651 TI - VEGF T-1498C polymorphism, a predictive marker of differentiation of colorectal adenocarcinomas in Japanese. AB - BACKGROUND: Previously, MDR1 T-129C polymorphism, encoding multidrug resistant transporter MDR1/P-glycoprotein, was reported to be predictive of poorly differentiated colorectal adenocarcinomas. Here, VEGF T-1498C, C-634G and C-7T polymorphisms, encoding vascular endothelial growth factor (VEGF), were investigated in terms of their association with differentiation grade. METHODS: VEGF genotypes were determined by TaqMan(R) MGB probe based polymerase chain reaction and evaluated were confirmed by direct sequencing in 36 Japanese patients. RESULTS: VEGF T-1498C, but not C-634G or C-7T, was predictive of poorly differentiated ones, and thereby a poor prognosis (p = 0.064 for genotype, p = 0.037 for allele), and this effect can be explained by that on VEGF expression. Treatment of a colorectal adenocarcinoma cell line, HCT-15, with sodium butyrate, a typical differentiating agent, resulted in an increase of alkaline phosphatase activity and MDR1 mRNA expression, but in a decrease of VEGF mRNA expression. The transfection of VEGF small interfering RNA (siRNA) induced the expression of MDR1 mRNA to 288-332% of the control level, whereas MDR1 siRNA had no effect on VEGF mRNA expression. CONCLUSIONS: VEGF T-1498C polymorphism is also a candidate marker predictive of poorly-differentiated colorectal adenocarcinomas, but further investigations with a large number of patients should be addressed to draw a conclusion. PMID- 18414652 TI - Lineage tracing of cardiac explant derived cells. AB - AIMS: Cultured cardiac explants produce a heterogeneous population of cells including a distinctive population of refractile cells described here as small round cardiac explant derived cells (EDCs). The aim of this study was to explore the source, morphology and cardiogenic potential of EDCs. METHODS: Transgenic MLC2v-Cre/ZEG, and actin-eGFP mice were used for lineage-tracing of EDCs in vitro and in vivo. C57B16 mice were used as cell transplant recipients of EDCs from transgenic hearts, as well as for the general characterisation of EDCs. The activation of cardiac-specific markers were analysed by: immunohistochemistry with bright field and immunofluorescent microscopy, electron microscopy, PCR and RT-PCR. Functional engraftment of transplanted cells was further investigated with calcium transient studies. RESULTS: Production of EDCs was highly dependent on the retention of blood-derived cells or factors in the cultured explants. These cells shared some characteristics of cardiac myocytes in vitro and survived engraftment in the adult heart in vivo. However, EDCs failed to differentiate into functional cardiac myocytes in vivo as demonstrated by the absence of stimulation-evoked intracellular calcium transients following transplantation into the peri-infarct zone. CONCLUSIONS: This study highlights that positive identification based upon one parameter alone such as morphology or immunofluorescene is not adequate to identify the source, fate and function of adult cardiac explant derived cells. PMID- 18414653 TI - Integration across Time Determines Path Deviation Discrimination for Moving Objects. AB - BACKGROUND: Human vision is vital in determining our interaction with the outside world. In this study we characterize our ability to judge changes in the direction of motion of objects-a common task which can allow us either to intercept moving objects, or else avoid them if they pose a threat. METHODOLOGY/PRINCIPAL FINDINGS: Observers were presented with objects which moved across a computer monitor on a linear path until the midline, at which point they changed their direction of motion, and observers were required to judge the direction of change. In keeping with the variety of objects we encounter in the real world, we varied characteristics of the moving stimuli such as velocity, extent of motion path and the object size. Furthermore, we compared performance for moving objects with the ability of observers to detect a deviation in a line which formed the static trace of the motion path, since it has been suggested that a form of static memory trace may form the basis for these types of judgment. The static line judgments were well described by a 'scale invariant' model in which any two stimuli which possess the same two-dimensional geometry (length/width) result in the same level of performance. Performance for the moving objects was entirely different. Irrespective of the path length, object size or velocity of motion, path deviation thresholds depended simply upon the duration of the motion path in seconds. CONCLUSIONS/SIGNIFICANCE: Human vision has long been known to integrate information across space in order to solve spatial tasks such as judgment of orientation or position. Here we demonstrate an intriguing mechanism which integrates direction information across time in order to optimize the judgment of path deviation for moving objects. PMID- 18414654 TI - Right isomerism of the brain in inversus viscerum mutant mice. AB - Left-right (L-R) asymmetry is a fundamental feature of higher-order neural function. However, the molecular basis of brain asymmetry remains unclear. We recently reported L-R asymmetry of hippocampal circuitry caused by differential allocation of N-methyl-D-aspartate receptor (NMDAR) subunit GluRepsilon2 (NR2B) in hippocampal synapses. Using electrophysiology and immunocytochemistry, here we analyzed the hippocampal circuitry of the inversus viscerum (iv) mouse that has a randomized laterality of internal organs. The iv mouse hippocampus lacks L-R asymmetry, it exhibits right isomerism in the synaptic distribution of the epsilon2 subunit, irrespective of the laterality of visceral organs. This independent right isomerism of the hippocampus is the first evidence that a distinct mechanism downstream of the iv mutation generates brain asymmetry. PMID- 18414655 TI - Optimal foraging predicts the ecology but not the evolution of host specialization in bacteriophages. AB - We explore the ability of optimal foraging theory to explain the observation among marine bacteriophages that host range appears to be negatively correlated with host abundance in the local marine environment. We modified Charnov's classic diet composition model to describe the ecological dynamics of the related generalist and specialist bacteriophages phiX174 and G4, and confirmed that specialist phages are ecologically favored only at high host densities. Our modified model accurately predicted the ecological dynamics of phage populations in laboratory microcosms, but had only limited success predicting evolutionary dynamics. We monitored evolution of attachment rate, the phenotype that governs diet breadth, in phage populations adapting to both low and high host density microcosms. Although generalist phiX174 populations evolved even broader diets at low host density, they did not show a tendency to evolve the predicted specialist foraging strategy at high host density. Similarly, specialist G4 populations were unable to evolve the predicted generalist foraging strategy at low host density. These results demonstrate that optimal foraging models developed to explain the behaviorally determined diets of predators may have only limited success predicting the genetically determined diets of bacteriophage, and that optimal foraging probably plays a smaller role than genetic constraints in the evolution of host specialization in bacteriophages. PMID- 18414657 TI - Analyses of in vivo interaction and mobility of two spliceosomal proteins using FRAP and BiFC. AB - U1-70K, a U1 snRNP-specific protein, and serine/arginine-rich (SR) proteins are components of the spliceosome and play critical roles in both constitutive and alternative pre-mRNA splicing. However, the mobility properties of U1-70K, its in vivo interaction with SR proteins, and the mobility of the U1-70K-SR protein complex have not been studied in any system. Here, we studied the in vivo interaction of U1-70K with an SR protein (SR45) and the mobility of the U1-70K/SR protein complex using bimolecular fluorescence complementation (BiFC) and fluorescence recovery after photobleaching (FRAP). Our results show that U1-70K exchanges between speckles and the nucleoplasmic pool very rapidly and that this exchange is sensitive to ongoing transcription and phosphorylation. BiFC analyses showed that U1-70K and SR45 interacted primarily in speckles and that this interaction is mediated by the RS1 or RS2 domain of SR45. FRAP analyses showed considerably slower recovery of the SR45/U1-70K complex than either protein alone indicating that SR45/U1-70K complexes remain in the speckles for a longer duration. Furthermore, FRAP analyses with SR45/U1-70K complex in the presence of inhibitors of phosphorylation did not reveal any significant change compared to control cells, suggesting that the mobility of the complex is not affected by the status of protein phosphorylation. These results indicate that U1-70K, like SR splicing factors, moves rapidly in the nucleus ensuring its availability at various sites of splicing. Furthermore, although it appears that U1-70K moves by diffusion its mobility is regulated by phosphorylation and transcription. PMID- 18414656 TI - Cryptococcal neuroradiological lesions correlate with severity during cryptococcal meningoencephalitis in HIV-positive patients in the HAART era. AB - Cryptococcal meningoencephalitis has an overall global mortality rate of 20% in AIDS patients despite antifungals. There is a need for additional means of precise assessment of disease severity. We thus studied the radiological brain images available from 62 HIV-positive patients with cryptococcocal meningoencephalitis to analyse the brain lesions associated with cryptococcosis in relationship with disease severity, and the respective diagnostic contribution of magnetic resonance (MR) versus computed tomography (CT). In this retrospective multicenter analysis, two neuroradiologists blindly reviewed the brain imaging. Prospectively acquired clinical and mycological data were available at baseline and during follow-up. Baseline images were abnormal on 92% of the MR scans contrasting with 53% of the CT scans. MR/CT cryptococcosis-related lesions included mass(es) (21%/9%), dilated perivascular spaces (46%/5%) and pseudocysts (8%/4%). The presence compared to absence of cryptococcosis-related lesions was significantly associated with high serum (78% vs. 42%, p = 0.008) and CSF (81% vs. 50%, p = 0.024) antigen titers, independently of neurological abnormalities. MR detected significantly more cryptococcosis-related lesions than CT for 17 patients who had had both investigations (76% vs. 24%, p = 0.005). In conclusion, MR appears more effective than CT for the evaluation of AIDS-associated cerebral cryptococcosis. Furthermore, brain imaging is an effective tool to assess the initial disease severity in this setting. Given this, we suggest that investigation for cryptococcosis-related lesions is merited, even in the absence of neurological abnormality, if a high fungal burden is suspected on the basis of high serum and/or CSF antigen titers. PMID- 18414658 TI - Establishing a cohort at high risk of HIV infection in South Africa: challenges and experiences of the CAPRISA 002 acute infection study. AB - OBJECTIVES: To describe the baseline demographic data, clinical characteristics and HIV-incidence rates of a cohort at high risk for HIV infection in South Africa as well as the challenges experienced in establishing and maintaining the cohort. METHODOLOGY/PRINCIPLE FINDINGS: Between August 2004 and May 2005 a cohort of HIV-uninfected women was established for the CAPRISA 002 Acute Infection Study, a natural history study of HIV-1 subtype C infection. Volunteers were identified through peer-outreach. The cohort was followed monthly to determine HIV infection rates and clinical presentation of early HIV infection. Risk reduction counselling and male and female condoms were provided. After screening 775 individuals, a cohort of 245 uninfected high-risk women was established. HIV prevalence at screening was 59.6% (95% CI: 55.9% to 62.8%) posing a challenge in accruing HIV-uninfected women. The majority of women (78.8%) were self-identified as sex-workers with a median of 2 clients per day. Most women (95%) reported more than one casual sexual partner in the previous 3 months (excluding clients) and 58.8% reported condom use in their last sexual encounter. Based on laboratory testing, 62.0% had a sexually transmitted infection at baseline. During 390 person-years of follow-up, 28 infections occurred yielding seroincidence rate of 7.2 (95% CI: 4.5 to 9.8) per 100 person-years. Despite the high mobility of this sex worker cohort retention rate after 2 years was 86.1%. High co-morbidity created challenges for ancillary care provision, both in terms of human and financial resources. CONCLUSIONS/SIGNIFICANCE: Challenges experienced were high baseline HIV-prevalence, lower than anticipated HIV-incidence and difficulties retaining participants. Despite challenges, we have successfully accrued this cohort of HIV-uninfected women with favourable retention, enabling us to study the natural history of HIV-1 during acute HIV-infection. Our experiences provide lessons for others establishing similar cohorts, which will be key for advancing the vaccine and prevention research agenda in resource-constrained settings. PMID- 18414659 TI - High-efficiency stem cell fusion-mediated assay reveals Sall4 as an enhancer of reprogramming. AB - Several methods allow reprogramming of differentiated somatic cells to embryonic stem cell-like cells. However, the process of reprogramming remains inefficient and the underlying molecular mechanisms are poorly understood. Here, we report the optimization of somatic cell fusion with embryonic stem cells in order to provide an efficient, quantitative assay to screen for factors that facilitate reprogramming. Following optimization, we achieved a reprogramming efficiency 15 590 fold higher than previous protocols. This allowed observation of cellular events during the reprogramming process. Moreover, we demonstrate that overexpression of the Spalt transcription factor, Sall4, which was previously identified as a regulator of embryonic stem cell pluripotency and early mouse development, can enhance reprogramming. The reprogramming activity of Sall4 is independent of an N-terminal domain implicated in recruiting the nucleosome remodeling and deacetylase corepressor complex, a global transcriptional repressor. These results indicate that improvements in reprogramming assays, including fusion assays, may allow the systematic identification and molecular characterization of enhancers of somatic cell reprogramming. PMID- 18414660 TI - Seasonal migration determined by a trade-off between predator avoidance and growth. AB - Migration is a common phenomenon in many organisms, terrestrial as well as aquatic, and considerable effort has been spent to understand the evolution of migratory behaviour and its consequences for population and community dynamics. In aquatic systems, studies on migration have mainly been focused on commercially important fish species, such as salmon and trout. However, seasonal mass migrations may occur also among other freshwater fish, e.g. in cyprinids that leave lakes and migrate into streams and wetlands in the fall and return back to the lake in spring. In a conceptual model, we hypothesized that this is an adaptive behaviour in response to seasonal changes in predation (P) and growth (G) and that migrating fish change habitat so as to minimise the ratio between predation mortality and growth rate (P/G). Estimates from bioenergetic modelling showed that seasonal changes in the ratio between predator consumption rate and prey growth rate followed the predictions from the conceptual model and also gave more precise predictions for the timing of the habitat change. By quantifying the migration of more than 1800 individually marked fish, we showed that actual migration patterns followed predictions with a remarkable accuracy, suggesting that migration patterns have evolved in response to seasonally fluctuating trade offs between predator avoidance and foraging gains. Thus, the conceptual model provides a mechanistic understanding to mass-migration in prey fish. Further, we also show that the dominant prey fish is actually absent from the lake during a major part of the year, which should have strong implications for the dynamics of the lake ecosystem through direct and indirect food-web interactions. PMID- 18414661 TI - A supplementary description of Cypridina mariae and rediagnosis of the genus Cylindroleberis (Ostracoda: Myodocopa: Cylindroleberididae). AB - The ostracod family Cylindroleberididae is based on the genus Cylindroleberis Brady, 1868, and has a complicated nomenclatural history. The type species of Cylindroleberis is Cypridina mariae Baird, 1850. Baird described only the carapace, which had been considered lost. Thus, there was no reference point for the concept C. mariae or the genus Cylindroleberis. Baird's material has now been found in the Natural History Museum, London, U.K., and is illustrated here. To clarify the taxonomic status of C. mariae and Cylindroleberis, specimens were obtained from near the type locality, and a supplementary description is presented. This includes description of appendages, particularly the first antenna and mandible, which contain important diagnostic characters. This supplementary description provides important information about C. mariae, allowing a revision of the genus Cylindroleberis, and establishing a framework for future biological research on this ostracod group. PMID- 18414662 TI - EWS/FLI mediates transcriptional repression via NKX2.2 during oncogenic transformation in Ewing's sarcoma. AB - EWS/FLI is a master regulator of Ewing's sarcoma formation. Gene expression studies in A673 Ewing's sarcoma cells have demonstrated that EWS/FLI downregulates more genes than it upregulates, suggesting that EWS/FLI, and/or its targets, function as transcriptional repressors. One critical EWS/FLI target, NKX2.2, is a transcription factor that contains both transcriptional activation and transcriptional repression domains, raising the possibility that it mediates portions of the EWS/FLI transcriptional signature. We now report that microarray analysis demonstrated that the transcriptional profile of NKX2.2 consists solely of downregulated genes, and overlaps with the EWS/FLI downregulated signature, suggesting that NKX2.2 mediates oncogenic transformation via transcriptional repression. Structure-function analysis revealed that the DNA binding and repressor domains in NKX2.2 are required for oncogenesis in Ewing's sarcoma cells, while the transcriptional activation domain is completely dispensable. Furthermore, blockade of TLE or HDAC function, two protein families thought to mediate the repressive function of NKX2.2, inhibited the transformed phenotype and reversed the NKX2.2 transcriptional profile in Ewing's sarcoma cells. Whole genome localization studies (ChIP-chip) revealed that a significant portion of the NKX2.2-repressed gene expression signature was directly mediated by NKX2.2 binding. These data demonstrate that the transcriptional repressive function of NKX2.2 is necessary, and sufficient, for the oncogenic phenotype of Ewing's sarcoma, and suggest a therapeutic approach to this disease. PMID- 18414663 TI - Phonological representations are unconsciously used when processing complex, non speech signals. AB - Neuroimaging studies of speech processing increasingly rely on artificial speech like sounds whose perceptual status as speech or non-speech is assigned by simple subjective judgments; brain activation patterns are interpreted according to these status assignments. The naive perceptual status of one such stimulus, spectrally-rotated speech (not consciously perceived as speech by naive subjects), was evaluated in discrimination and forced identification experiments. Discrimination of variation in spectrally-rotated syllables in one group of naive subjects was strongly related to the pattern of similarities in phonological identification of the same stimuli provided by a second, independent group of naive subjects, suggesting either that (1) naive rotated syllable perception involves phonetic-like processing, or (2) that perception is solely based on physical acoustic similarity, and similar sounds are provided with similar phonetic identities. Analysis of acoustic (Euclidean distances of center frequency values of formants) and phonetic similarities in the perception of the vowel portions of the rotated syllables revealed that discrimination was significantly and independently influenced by both acoustic and phonological information. We conclude that simple subjective assessments of artificial speech like sounds can be misleading, as perception of such sounds may initially and unconsciously utilize speech-like, phonological processing. PMID- 18414664 TI - Sialoadhesin expressed on IFN-induced monocytes binds HIV-1 and enhances infectivity. AB - BACKGROUND: HIV-1 infection dysregulates the immune system and alters gene expression in circulating monocytes. Differential gene expression analysis of CD14(+) monocytes from subjects infected with HIV-1 revealed increased expression of sialoadhesin (Sn, CD169, Siglec 1), a cell adhesion molecule first described in a subset of macrophages activated in chronic inflammatory diseases. METHODOLOGY/PRINCIPAL FINDINGS: We analyzed sialoadhesin expression on CD14(+) monocytes by flow cytometry and found significantly higher expression in subjects with elevated viral loads compared to subjects with undetectable viral loads. In cultured CD14(+) monocytes isolated from healthy individuals, sialoadhesin expression was induced by interferon-alpha and interferon-gamma but not tumor necrosis factor-alpha. Using a stringent binding assay, sialoadhesin-expressing monocytes adsorbed HIV-1 through interaction with the sialic acid residues on the viral envelope glycoprotein gp120. Furthermore, monocytes expressing sialoadhesin facilitated HIV-1 trans infection of permissive cells, which occurred in the absence of monocyte self-infection. CONCLUSIONS/SIGNIFICANCE: Increased sialoadhesin expression on CD14(+) monocytes occurred in response to HIV-1 infection with maximum expression associated with high viral load. We show that interferons induce sialoadhesin in primary CD14(+) monocytes, which is consistent with an antiviral response during viremia. Our findings suggest that circulating sialoadhesin-expressing monocytes are capable of binding HIV-1 and effectively delivering virus to target cells thereby enhancing the distribution of HIV-1. Sialoadhesin could disseminate HIV-1 to viral reservoirs during monocyte immunosurveillance or migration to sites of inflammation and then facilitate HIV 1 infection of permissive cells. PMID- 18414665 TI - High genetic differentiation between the M and S molecular forms of Anopheles gambiae in Africa. AB - BACKGROUND: Anopheles gambiae, a major vector of malaria, is widely distributed throughout sub-Saharan Africa. In an attempt to eliminate infective mosquitoes, researchers are trying to develop transgenic strains that are refractory to the Plasmodium parasite. Before any release of transgenic mosquitoes can be envisaged, we need an accurate picture of the differentiation between the two molecular forms of An. gambiae, termed M and S, which are of uncertain taxonomic status. METHODOLOGY/PRINCIPAL FINDINGS: Insertion patterns of three transposable elements (TEs) were determined in populations from Benin, Burkina Faso, Cameroon, Ghana, Ivory Coast, Madagascar, Mali, Mozambique, Niger, and Tanzania, using Transposon Display, a TE-anchored strategy based on Amplified Fragment Length Polymorphism. The results reveal a clear differentiation between the M and S forms, whatever their geographical origin, suggesting an incipient speciation process. CONCLUSIONS/SIGNIFICANCE: Any attempt to control the transmission of malaria by An. gambiae using either conventional or novel technologies must take the M/S genetic differentiation into account. In addition, we localized three TE insertion sites that were present either in every individual or at a high frequency in the M molecular form. These sites were found to be located outside the chromosomal regions that are suspected of involvement in the speciation event between the two forms. This suggests that these chromosomal regions are either larger than previously thought, or there are additional differentiated genomic regions interspersed with undifferentiated regions. PMID- 18414666 TI - Cerebrospinal fluid viral load and intrathecal immune activation in individuals infected with different HIV-1 genetic subtypes. AB - BACKGROUND: HIV-1 exhibits a high degree of genetic diversity and is presently divided into 3 distinct HIV-1 genetic groups designated major (M), non-M/non-O (N) and outlier (O). Group M, which currently comprises 9 subtypes (A-D, F-H, J and K), at least 34 circulating recombinant forms (CRFs) and several unique recombinant forms (URFs) is responsible for most of the HIV-1 epidemic. Most of the current knowledge of HIV-1 central nervous system (CNS) infection is based on subtype B. However, subtypes other than subtype B account for the majority of global HIV-1 infections. Therefore, we investigated whether subtypes have any influence on cerebrospinal fluid (CSF) markers of HIV-1 CNS infection. METHODOLOGY/PRINCIPAL FINDINGS: CSF HIV-1 RNA, CSF neopterin and CSF white blood cell (WBC) count were measured in patients infected with different HIV-1 subtypes. Using multivariate regression analysis, no differences in the CSF WBC count, neopterin and viral load were found between various HIV-1 subtypes. CONCLUSIONS: We did not find any subtype-dependent differences in the markers evaluated in this study. PMID- 18414668 TI - Activation of interleukin-32 pro-inflammatory pathway in response to influenza A virus infection. AB - BACKGROUND: Interleukin (IL)-32 is a recently described pro-inflammatory cytokine that has been reported to be induced by bacteria treatment in culture cells. Little is known about IL-32 production by exogenous pathogens infection in human individuals. METHODS AND FINDINGS: In this study, we found that IL-32 level was increased by 58.2% in the serum samples from a cohort of 108 patients infected by influenza A virus comparing to that of 115 healthy individuals. Another pro inflammatory factor cyclooxygenase (COX)-2-associated prostaglandin E2 was also upregulated by 2.7-fold. Expression of IL-32 in influenza A virus infected A549 human lung epithelial cells was blocked by either selective COX-2 inhibitor NS398 or Aspirin, a known anti-inflammatory drug, indicating IL-32 was induced through COX-2 in the inflammatory cascade. Interestingly, we found that COX-2-associate PGE(2) production activated by influenza virus infection was significantly suppressed by over-expression of IL-32 but increased by IL-32-specific siRNA, suggesting there was a feedback mechanism between IL-32 and COX-2. CONCLUSIONS: IL-32 is induced by influenza A virus infection via COX-2 in the inflammatory cascade. Our results provide that IL-32 is a potential target for anti inflammatory medicine screening. PMID- 18414667 TI - Immature cryopreserved ovary restores puberty and fertility in mice without alteration of epigenetic marks. AB - BACKGROUND: Progress in oncology could improve survival rate in children, but would probably lead to impaired fertility and puberty. In pre-pubertal girls, the only therapeutic option is the cryopreservation of one ovary. Three births have been reported after reimplantation of cryopreserved mature ovary. Conversely, reimplantation of ovary preserved before puberty (defined as immature ovary) has never been performed in humans. METHODOLOGY/PRINCIPAL FINDINGS: In order to analyze ovarian function, we performed transplantation using fresh or cryopreserved immature grafts in pre-pubertal or adult mice. Puberty as well as cyclic hormonal activity was restored. All follicle populations were present although a significant reduction in follicle density was observed with or without cryopreservation. Although fertility was restored, the graft is of limited life span. Because ex vivo ovary manipulation and cryopreservation procedure, the status of genomic imprinting was investigated. Methylation status of the H19 and Lit1 Imprinting Control Regions in kidney, muscle and tongue of offsprings from grafted mice does not show significant alteration when compared to those of unoperated mice. CONCLUSIONS/SIGNIFICANCE: These results demonstrate that immature ovarian grafting can restore spontaneous puberty and fertility. However, these data suggest that follicle depletion leads to premature ovarian failure. This study addresses the very important epigenetics issue, and provides valuable information to the study of ovarian transplantation suggesting that these procedures do not perturb normal epigenetics marks. These results are highly relevant to the reimplantation question of immature cortex in women. PMID- 18414669 TI - Does sex trade with violence among genotypes in Drosophila melanogaster? AB - The evolutionary forces shaping the ability to win competitive interactions, such as aggressive encounters, are still poorly understood. Given a fitness advantage for competitive success, variance in aggressive and sexual display traits should be depleted, but a great deal of variation in these traits is consistently found. While life history tradeoffs have been commonly cited as a mechanism for the maintenance of variation, the variability of competing strategies of conspecifics may mean there is no single optimum strategy. We measured the genetically determined outcomes of aggressive interactions, and the resulting effects on mating success, in a panel of diverse inbred lines representing both natural variation and artificially selected genotypes. Males of one genotype which consistently lost territorial encounters with other genotypes were nonetheless successful against males that were artificially selected for supernormal aggression and dominated all other lines. Intransitive patterns of territorial success could maintain variation in aggressive strategies if there is a preference for territorial males. Territorial success was not always associated with male mating success however and females preferred 'winners' among some male genotypes, and 'losers' among other male genotypes. This suggests that studying behaviour from the perspective of population means may provide limited evolutionary and genetic insight. Overall patterns of competitive success among males and mating transactions between the sexes are consistent with mechanisms proposed for the maintenance of genetic variation due to nonlinear outcomes of competitive interactions. PMID- 18414670 TI - Functional significance of allelic variation at methuselah, an aging gene in Drosophila. AB - BACKGROUND: Longevity and age-specific patterns of mortality are complex traits that vary within and among taxa. Multiple candidate genes for aging have been identified in model systems by extended longevity mutant phenotypes, including the G-protein coupled receptor methuselah (mth) in D. melanogaster. These genes offer important insights into the mechanisms of lifespan determination and have been major targets of interest in the biology of aging. However, it is largely unknown whether these genes contribute to genetic variance for lifespan in natural populations, and consequently contribute to lifespan evolution. METHODOLOGY/PRINCIPLE FINDINGS: For a gene to contribute to genetic variance for a particular trait, it must meet two criteria: natural allelic variation and functional differences among variants. Previous work showed that mth varies significantly among wild populations; here we assess the functional significance of wild-derived mth alleles on lifespan, fecundity and stress resistance using a quantitative complementation scheme. Our results demonstrate that mth alleles segregating in nature have a functional effect on all three traits. CONCLUSIONS/SIGNIFICANCE: These results suggest that allelic variation at mth contributes to observed differences in lifespan and correlated phenotypes in natural populations, and that evaluation of genetic diversity at candidate genes for aging can be a fruitful approach to identifying loci contributing to lifespan evolution. PMID- 18414671 TI - Vpr14-88-Apobec3G fusion protein is efficiently incorporated into Vif-positive HIV-1 particles and inhibits viral infection. AB - BACKGROUND: APOBEC3G (A3G), a deoxycytidine deaminase, is a potent host antiviral factor that can restrict HIV-1 infection. During Vif-negative HIV-1 replication, A3G is incorporated into HIV-1 particles, induces mutations in reverse transcribed viral DNA and inhibits reverse transcription. However, HIV-1 Vif counteracts A3G's activities by inducing its degradation and by blocking its incorporation into HIV-1 particles. Thus, it is interesting to elucidate a mechanism that would allow A3G to escape the effects of Vif in order to rescue its potent antiviral activity and to provide a possible novel therapeutic strategy for treating HIV-1 infection. METHODS AND FINDINGS: In this study, we generated an R88-A3G fusion protein by fusing A3G to a virion-targeting polypeptide (R14-88) derived from HIV-1 Vpr protein and compared its antiviral effects relative to those of HA-tagged native A3G (HA-A3G). Our study showed that transient expression of the R88-A3G fusion protein in both Vif(-) and Vif(+) HIV 1 producing cells drastically inhibited viral infection in HeLa-CD4-CCR5-cells, CD4(+) C8166 T cells and human primary PBMCs. Moreover, we established CD4(+) C8166 T cell lines that stably express either R88-A3G or HA-A3G by transduction with VSV-G-pseudotyped lentiviral vector that harbor expression cassettes for R88 A3G or HA-A3G, respectively, and tested their susceptibility to Vif(+) HIV-1 infection. Our results clearly reveal that expression of R88-A3G in transduced CD4(+) C8166 cells significantly blocked Vif(+) HIV-1 infection. In an attempt to understand the mechanism underlying the antiviral activity of R88-A3G, we demonstrated that R88-A3G was efficiently incorporated into viral particles in the presence of Vif. Moreover, PCR analysis revealed that R88-A3G significantly inhibited viral cDNA synthesis during the early stage of Vif(+) virus infection. CONCLUSIONS: Our results clearly indicate that R88 delivers A3G into Vif(+) HIV-1 particles and inhibits infectivity and spread of the virions among CD4(+) T cells. This study provides evidence for an effective strategy to modify a host protein with innate anti-HIV-1 activity and rescue its potent anti-HIV potential in the presence of Vif. Further characterization and optimization of this system may lead to the development of an effective therapeutic approach against HIV-1 infection. PMID- 18414672 TI - Restricted expression of Epstein-Barr virus latent genes in murine B cells derived from embryonic stem cells. AB - BACKGROUND: Several human malignancies are associated with Epstein-Barr virus (EBV) and more than 95% of the adult human population carries this virus lifelong. EBV efficiently infects human B cells and persists in this cellular compartment latently. EBV-infected B cells become activated and growth transformed, express a characteristic set of viral latent genes, and acquire the status of proliferating lymphoblastoid cell lines in vitro. Because EBV infects only primate cells, it has not been possible to establish a model of infection in immunocompetent rodents. Such a model would be most desirable in order to study EBV's pathogenesis and latency in a suitable and amenable host. METHODOLOGY/PRINCIPAL FINDINGS: We stably introduced recombinant EBV genomes into mouse embryonic stem cells and induced their differentiation to B cells in vitro to develop the desired model. In vitro differentiated murine B cells maintained the EBV genomes but expression of viral genes was restricted to the latent membrane proteins (LMPs). In contrast to human B cells, EBV's nuclear antigens (EBNAs) were not expressed detectably and growth transformed murine B cells did not arise in vitro. Aberrant splicing and premature termination of EBNA mRNAs most likely prevented the expression of EBNA genes required for B-cell transformation. CONCLUSIONS/SIGNIFICANCE: Our findings indicate that fundamental differences in gene regulation between mouse and man might block the route towards a tractable murine model for EBV. PMID- 18414673 TI - Pathways to injury in chronic pancreatitis: decoding the role of the high-risk SPINK1 N34S haplotype using meta-analysis. AB - BACKGROUND: The complex interactions between recurrent trypsin-mediated pancreatic injury, alcohol-associated pancreatic injury and SPINK1 polymorphisms in chronic pancreatitis (CP) are undefined. We hypothesize that CP occurs as a result of multiple pathological mechanisms (pathways) that are initiated by different metabolic or environmental factors (etiologies) and may be influenced differentially by downstream genetic risk factors. We tested this hypothesis by evaluating the differences in effect size of the high risk SPINK1 N34S haplotype on CP from multiple etiologies after combining clinical reports of SPINK1 N34S frequency using meta-analysis. METHODS AND FINDINGS: The Pubmed and the Embase databases were reviewed. We studied 24 reports of SPINK1 N34S in CP (2,421 cases, 4,857 controls) using reported etiological factors as surrogates for pathways and multiple meta-analyses to determine the differential effects of SPINK1 N34S between alcoholic and non-alcoholic etiologies. Using estimates of between-study heterogeneity, we sub-classified our 24 studies into four specific clusters. We found that SPINK1 N34S is strongly associated with CP overall (OR 11.00; 95% CI: 7.59-15.93), but the effect of SPINK1 N34S in alcoholic CP (OR 4.98, 95% CI: 3.16 7.85) was significantly smaller than in idiopathic CP (OR 14.97, 95% C.I. = 9.09 24.67) or tropical CP (OR 19.15, 95% C.I. = 8.83-41.56). Studies analyzing familial CP showed very high heterogeneity suggestive of a complex etiology with an I(2) = 80.95%. CONCLUSION: The small effect of SPINK1 N34S in alcoholic subjects suggests that CP is driven through a different pathway that is largely trypsin-independent. The results also suggest that large effect sizes of SPINK1 N34S in small candidate gene studies in CP may be related to a mixture of multiple etiologic pathways leading to the same clinical endpoint. PMID- 18414674 TI - Cyclic and sleep-like spontaneous alternations of brain state under urethane anaesthesia. AB - BACKGROUND: Although the induction of behavioural unconsciousness during sleep and general anaesthesia has been shown to involve overlapping brain mechanisms, sleep involves cyclic fluctuations between different brain states known as active (paradoxical or rapid eye movement: REM) and quiet (slow-wave or non-REM: nREM) stages whereas commonly used general anaesthetics induce a unitary slow-wave brain state. METHODOLOGY/PRINCIPAL FINDINGS: Long-duration, multi-site forebrain field recordings were performed in urethane-anaesthetized rats. A spontaneous and rhythmic alternation of brain state between activated and deactivated electroencephalographic (EEG) patterns was observed. Individual states and their transitions resembled the REM/nREM cycle of natural sleep in their EEG components, evolution, and time frame ( approximately 11 minute period). Other physiological variables such as muscular tone, respiration rate, and cardiac frequency also covaried with forebrain state in a manner identical to sleep. The brain mechanisms of state alternations under urethane also closely overlapped those of natural sleep in their sensitivity to cholinergic pharmacological agents and dependence upon activity in the basal forebrain nuclei that are the major source of forebrain acetylcholine. Lastly, stimulation of brainstem regions thought to pace state alternations in sleep transiently disrupted state alternations under urethane. CONCLUSIONS/SIGNIFICANCE: Our results suggest that urethane promotes a condition of behavioural unconsciousness that closely mimics the full spectrum of natural sleep. The use of urethane anaesthesia as a model system will facilitate mechanistic studies into sleep-like brain states and their alternations. In addition, it could also be exploited as a tool for the discovery of new molecular targets that are designed to promote sleep without compromising state alternations. PMID- 18414676 TI - Seasonal activity budget of adult baltic ringed seals. AB - Although ringed seals are important components in oceanic and fresh water ecosystems at high latitudes, little is known about how they exploit these harsh environments. Seasonal activity and diving behaviour of 19 adult Baltic ringed seals were studied by satellite telemetry. We elaborated an activity budget for ten months of the year, extending over the period from moult to the breeding season. Seals from three main regions showed explicit site fidelity and the distributions of animals tagged from different areas did not overlap, suggesting separate stocks. Both the mean duration and the mean depth of dives peaked in June and July. Seals spent 70% (females) to 85% (males) of their time diving in June and July which decreased to 50% in late autumn. Less than one percent of dives exceeded 10 min in females, while 10% of male dives lasted longer than 10 min in June to September. Less than one percent of dives lasted for more than 25 min. Both females and males were most active during day time and hauled out predominantly during the night. Activity patterns during the summer are suggested to be correlated to energy accumulation and prey availability. The information on seasonal activity budget is crucial for developing population energetic models where interactions between ringed seals and other trophic levels can be evaluated. PMID- 18414675 TI - High-resolution labeling and functional manipulation of specific neuron types in mouse brain by Cre-activated viral gene expression. AB - We describe a method that combines Cre-recombinase knockin mice and viral mediated gene transfer to genetically label and functionally manipulate specific neuron types in the mouse brain. We engineered adeno-associated viruses (AAVs) that express GFP, dsRedExpress, or channelrhodopsin (ChR2) upon Cre/loxP recombination-mediated removal of a transcription-translation STOP cassette. Fluorescent labeling was sufficient to visualize neuronal structures with synaptic resolution in vivo, and ChR2 expression allowed light activation of neuronal spiking. The structural dynamics of a specific class of neocortical neuron, the parvalbumin-containing (Pv) fast-spiking GABAergic interneuron, was monitored over the course of a week. We found that although the majority of Pv axonal boutons were stable in young adults, bouton additions and subtractions on axonal shafts were readily observed at a rate of 10.10% and 9.47%, respectively, over 7 days. Our results indicate that Pv inhibitory circuits maintain the potential for structural re-wiring in post-adolescent cortex. With the generation of an increasing number of Cre knockin mice and because viral transfection can be delivered to defined brain regions at defined developmental stages, this strategy represents a general method to systematically visualize the structure and manipulate the function of different cell types in the mouse brain. PMID- 18414677 TI - Methylphenidate decreased the amount of glucose needed by the brain to perform a cognitive task. AB - The use of stimulants (methylphenidate and amphetamine) as cognitive enhancers by the general public is increasing and is controversial. It is still unclear how they work or why they improve performance in some individuals but impair it in others. To test the hypothesis that stimulants enhance signal to noise ratio of neuronal activity and thereby reduce cerebral activity by increasing efficiency, we measured the effects of methylphenidate on brain glucose utilization in healthy adults. We measured brain glucose metabolism (using Positron Emission Tomography and 2-deoxy-2[18F]fluoro-D-glucose) in 23 healthy adults who were tested at baseline and while performing an accuracy-controlled cognitive task (numerical calculations) given with and without methylphenidate (20 mg, oral). Sixteen subjects underwent a fourth scan with methylphenidate but without cognitive stimulation. Compared to placebo methylphenidate significantly reduced the amount of glucose utilized by the brain when performing the cognitive task but methylphenidate did not affect brain metabolism when given without cognitive stimulation. Whole brain metabolism when the cognitive task was given with placebo increased 21% whereas with methylphenidate it increased 11% (50% less). This reflected both a decrease in magnitude of activation and in the regions activated by the task. Methylphenidate's reduction of the metabolic increases in regions from the default network (implicated in mind-wandering) was associated with improvement in performance only in subjects who activated these regions when the cognitive task was given with placebo. These results corroborate prior findings that stimulant medications reduced the magnitude of regional activation to a task and in addition document a "focusing" of the activation. This effect may be beneficial when neuronal resources are diverted (i.e., mind-wandering) or impaired (i.e., attention deficit hyperactivity disorder), but it could be detrimental when brain activity is already optimally focused. This would explain why methylphenidate has beneficial effects in some individuals and contexts and detrimental effects in others. PMID- 18414678 TI - Unique environmental effects on physical activity participation: a twin study. AB - BACKGROUND: The health benefits of regular physical activity are well established. However, the relative contribution of heritable and environmental factors to physical activity participation remains controversial. Using a cut point of 60 minutes of total activity per week, data from the GenomEUtwin project revealed consistent genetic influence on physical activity participation in 37,051 twin pairs from seven countries. We hypothesized that the heritability of physical activity participation would be attenuated using the CDC/ACSM recommended minimum threshold of 150 minutes of moderate intensity activity per week. METHODS: Data were obtained from 1,389 twin pairs from the community-based University of Washington Twin Registry. Twin similarity in physical activity participation using both cut-points was analyzed using tetrachoric correlations and structural equation modeling in all same-sex pairs. RESULTS: Correlations were higher in monozygotic (r(MZ) = 0.43, 95% CI = 0.33-0.54) than dizygotic pairs (r(DZ) = 0.30, 95% CI = 0.12-0.47) using the 60 minute cut-point. However, differences were attenuated using the 150 minute standard (r(MZ) = 0.30, 95% CI = 0.20-0.40; r(DZ) = 0.25, 95% CI = 0.07-0.42). Using the lower cut-point, the best fitting model of twin resemblance only included additive genetics and unique environment, with a heritability of 45%. In contrast, using the higher threshold, the best fitting model included the common and unique environment, with the unique environment contributing 72% of the variance. CONCLUSION: Unique environment factors provide the strongest influence on physical activity participation at levels recommended for health benefits. PMID- 18414679 TI - The SIRT1 deacetylase suppresses intestinal tumorigenesis and colon cancer growth. AB - Numerous longevity genes have been discovered in model organisms and altering their function results in prolonged lifespan. In mammals, some have speculated that any health benefits derived from manipulating these same pathways might be offset by increased cancer risk on account of their propensity to boost cell survival. The Sir2/SIRT1 family of NAD(+)-dependent deacetylases is proposed to underlie the health benefits of calorie restriction (CR), a diet that broadly suppresses cancer in mammals. Here we show that CR induces a two-fold increase SIRT1 expression in the intestine of rodents and that ectopic induction of SIRT1 in a beta-catenin-driven mouse model of colon cancer significantly reduces tumor formation, proliferation, and animal morbidity in the absence of CR. We show that SIRT1 deacetylates beta-catenin and suppresses its ability to activate transcription and drive cell proliferation. Moreover, SIRT1 promotes cytoplasmic localization of the otherwise nuclear-localized oncogenic form of beta-catenin. Consistent with this, a significant inverse correlation was found between the presence of nuclear SIRT1 and the oncogenic form of beta-catenin in 81 human colon tumor specimens analyzed. Taken together, these observations show that SIRT1 suppresses intestinal tumor formation in vivo and raise the prospect that therapies targeting SIRT1 may be of clinical use in beta-catenin-driven malignancies. PMID- 18414680 TI - Kidney injury molecule-1 is a phosphatidylserine receptor that confers a phagocytic phenotype on epithelial cells. AB - Following injury, the clearance of apoptotic and necrotic cells is necessary for mitigation and resolution of inflammation and tissue repair. In addition to macrophages, which are traditionally assigned to this task, neighboring epithelial cells in the affected tissue are postulated to contribute to this process. Kidney injury molecule-1 (KIM-1 or TIM-1) is an immunoglobulin superfamily cell-surface protein not expressed by cells of the myeloid lineage but highly upregulated on the surface of injured kidney epithelial cells. Here we demonstrate that injured kidney epithelial cells assumed attributes of endogenous phagocytes. Confocal images confirm internalization of apoptotic bodies within KIM-1-expressing epithelial cells after injury in rat kidney tubules in vivo. KIM 1 was directly responsible for phagocytosis in cultured primary rat tubule epithelial cells and also porcine and canine epithelial cell lines. KIM-1 was able to specifically recognize apoptotic cell surface-specific epitopes phosphatidylserine, and oxidized lipoproteins, expressed by apoptotic tubular epithelial cells. Thus, KIM-1 is the first nonmyeloid phosphatidylserine receptor identified to our knowledge that transforms epithelial cells into semiprofessional phagocytes. PMID- 18414681 TI - Mechanisms of an autoimmunity syndrome in mice caused by a dominant mutation in Aire. AB - Homozygous loss-of-function mutations in AIRE cause autoimmune polyglandular syndrome type 1 (APS 1), which manifests in a classic triad of hypoparathyroidism, adrenal insufficiency, and candidiasis. Interestingly, a kindred with a specific G228W AIRE variant presented with an autosomal dominant autoimmune phenotype distinct from APS 1. We utilized a novel G228W-knockin mouse model to show that this variant acted in a dominant-negative manner to cause a unique autoimmunity syndrome. In addition, the expression of a large number of Aire-regulated thymic antigens was partially inhibited in these animals, demonstrating the importance of quantitative changes in thymic antigen expression in determining organ-specific autoimmunity. Furthermore, the dominant-negative effect of the G228W variant was exerted through recruitment of WT Aire away from active sites of transcription in the nucleus of medullary thymic epithelial cells in vivo. Together, these results may demonstrate a mechanism by which autoimmune predisposition to phenotypes distinct from APS 1 can be mediated in a dominant negative fashion by Aire. PMID- 18414682 TI - Diminished Ret expression compromises neuronal survival in the colon and causes intestinal aganglionosis in mice. AB - Mutations in the RET gene are the primary cause of Hirschsprung disease (HSCR), or congenital intestinal aganglionosis. However, how RET malfunction leads to HSCR is not known. It has recently been shown that glial cell line-derived neurotrophic factor (GDNF) family receptor alpha1 (GFRalpha1), which binds to GDNF and activates RET, is essential for the survival of enteric neurons. In this study, we investigated Ret regulation of enteric neuron survival and its potential involvement in HSCR. Conditional ablation of Ret in postmigratory enteric neurons caused widespread neuronal death in the colon, which led to colonic aganglionosis. To further examine this finding, we generated a mouse model for HSCR by reducing Ret expression levels. These mice recapitulated the genetic and phenotypic features of HSCR and developed colonic aganglionosis due to impaired migration and successive death of enteric neural crest-derived cells. Death of enteric neurons was also induced in the colon, where reduction of Ret expression was induced after the period of enteric neural crest cell migration, indicating that diminished Ret expression directly affected the survival of colonic neurons. Thus, enteric neuron survival is sensitive to RET dosage, and cell death is potentially involved in the etiology of HSCR. PMID- 18414683 TI - ROCK1 mediates leukocyte recruitment and neointima formation following vascular injury. AB - Although Rho-associated kinase (ROCK) activity has been implicated in cardiovascular diseases, the tissue- and isoform-specific roles of ROCKs in the vascular response to injury are not known. To address the role of ROCKs in this process, we generated haploinsufficient Rock1 (Rock1(+/-)) and Rock2 (Rock2(+/-)) mice and performed carotid artery ligations. Following this intervention, we found reduced neointima formation in Rock1(+/-) mice compared with that of WT or Rock2(+/-) mice. This correlated with decreased vascular smooth muscle cell proliferation and survival, decreased levels proinflammatory adhesion molecule expression, and reduced leukocyte infiltration. In addition, thioglycollate induced peritoneal leukocyte recruitment and accumulation were substantially reduced in Rock1(+/-) mice compared with those of WT and Rock2(+/-) mice. To determine the role of leukocyte-derived ROCK1 in neointima formation, we performed reciprocal bone marrow transplantation (BMT) in WT and Rock1(+/-) mice. Rock1(+/-) to WT BMT led to reduced neointima formation and leukocyte infiltration following carotid ligation compared with those of WT to WT BMT. In contrast, WT to Rock1(+/-) BMT resulted in increased neointima formation. These findings indicate that ROCK1 in BM-derived cells mediates neointima formation following vascular injury and suggest that ROCK1 may represent a promising therapeutic target in vascular inflammatory diseases. PMID- 18414684 TI - Serotype-dependent packaging of large genes in adeno-associated viral vectors results in effective gene delivery in mice. AB - Vectors derived from adeno-associated virus (AAV) are promising for human gene therapy, including treatment for retinal blindness. One major limitation of AAVs as vectors is that AAV cargo capacity has been considered to be restricted to 4.7 kb. Here we demonstrate that vectors with an AAV5 capsid (i.e., rAAV2/5) incorporated up to 8.9 kb of genome more efficiently than 6 other serotypes tested, independent of the efficiency of the rAAV2/5 production process. Efficient packaging of the large murine Abca4 and human MYO7A and CEP290 genes, which are mutated in common blinding diseases, was obtained, suggesting that this packaging efficiency is independent of the specific sequence packaged. Expression of proteins of the appropriate size and function was observed following transduction with rAAV2/5 carrying large genes. Intraocular administration of rAAV2/5 encoding ABCA4 resulted in protein localization to rod outer segments and significant and stable morphological and functional improvement of the retina in Abca4(-/-) mice. This use of rAAV2/5 may be a promising therapeutic strategy for recessive Stargardt disease, the most common form of inherited macular degeneration. The possibility of packaging large genes in AAV greatly expands the therapeutic potential of this vector system. PMID- 18414686 TI - [Development and application of a two-step MacELISA for the early diagnosis of hemorrhagic fever with renal syndrome]. AB - OBJECTIVE: To develop and improve a MacELISA method for the early diagnosis of hemorrhagic fever with renal syndrome (HFRS) with simplified operation procedure. METHODS: The nucleic proteins of hantavirus were labeled with horse raddish peroxidase (HRP) and used as detection antigens. A two-step MacELISA based HRP conjugated antigen was established and the detection sensitivity and specificity were compared with commonly used three-step MacELISA. RESULTS: This method could be used to detect hantanvirus specific IgM with high sensitivity and specificity from human patient serum. There was not significant difference from commonly used three-step MacELISA and the sensitivity and specificity were 100%. CONCLUSION: This method is simple, sensitive and rapid in operation, and therefore could be used for the early diagnosis of HFRS. PMID- 18414685 TI - [Molecular characterization of hantavirus Zhejiang isolate ZT10 strain from M. fartis]. AB - OBJECTIVE: To learn about the complete genomic sequence of the Seoul virus strain ZT10 isolated from M. fartis. METHODS: The total RNA was extracted from the infected Vero E6 cells and amplified by RT-PCR. The purified PCR products were cloned into T-vector and sequenced. RESULTS: The results demonstrated that the complete genome of ZT10 was comprised of L(6530), M(3651) and S(1753) segments which encoded 2151-1133 and 429 amino acids respectively. CONCLUSION: Analysis of sequence revealed that the ZT10 belonged to Seoul virus. The nucleotide sequence identity of the M gene with Seoul virus was 84.0%-96.3%. The identity with Hantan vrisu (Prospect Hill virus, Tula virus) isolated from M. fartis was 57.5%-60.9%. The sequence identity of the S gene with Seoul virus was 87.9%-96.0% at nucleotide level and 96.9%-97.9% at amino acid level. PMID- 18414687 TI - [Cloning and sequence analysis of envelope glycoprotein G2 gene of hantavirus in Shandong province]. AB - OBJECTIVE: To construct the cloning vector of glycoprotein G2 gene of hantavirus (HV), to analyze the sequence of G2 gene by the phylogenetic tree, and to study the differences among glycoprotein G2 genes from the world around. METHODS: Envelope glycoprotein G2 gene was amplified from four specimens of Shandong province by RT-PCR, and the product recombined into the PMD-18T vector. The clones that carry the G2 gene were identified. After sequencing, the gene sequence was handled with the software DNASTAR, compared with 24 strains worldwide and the phylogenetic tree was drawn. RESULTS: HV G2 gene was amplified by RT-PCR from 4 specimens, named GM04-38.G2, ZB8.G2, JUN5-14.G2, RCH5.G2, respectively. The map of the phylogenetic tree showed that all the 4 strains belonged to SEO-type hantavirus. The analysis of the sequence showed that all the four HV strains had the highest rates of homology with Z37 strain. The sequence homology of SEO-type HV strains was from 82.3% to 99.8%. CONCLUSION: The four cloning vectors containing the glycoprotein G2 genes were successfully constructed. Envelope glycoprotein G2 gene of four specimens from Shandong province had high homology rates. PMID- 18414689 TI - [Genotype and sequence analysis on G2 segments of hantavirus from HFRS patients in Hebei Province]. AB - OBJECTIVE: To know the genotype and subtype of hantavirus (HV) which infected persons in Hebei province. METHODS: According to G2 coding region of 76-118 and R22 strains, specific type primers were designed to detect and identity the types of HV in HFRS patients' sera with RT-nested PCR. Nucleotides were assayed from partial products after purification and reclaim. Then, gene analysis was done with DNAStar package. RESULTS: 17 out of 69 positive serum specimens were successfully detected by RT-PCR and the detection rate was 24.64%, among which, or= 14 days were 0. 17 positive specimens were all belonged to SEO. The nucleotide homology of 9 typical specimens was 92.0%-100%. Between HeB7 and other 8 specimens was 92%-95%, and they belonged to different subtypes. When HeB7 compared with R22 strain, it was 97.7%. HeB7 and R22 belonged to S1 subtype. The 8 specimens except HeB7 was 95.7% 100% and they all belonged to S3 subtype. When compared with 76-118 strain, 9 specimens' nucleotide homology was only 70.3%-72.7%, belonged to different type. CONCLUSION: SEO was the major type of HV from HFRS patients in Hebei province, S3 was the major subtype and S1 was also existed. In a certain area, the HV which belonged to the same type was correspondingly conservative, and had the characteristic of regional stability. PMID- 18414690 TI - [Construction and immunological evaluation of recombinant adenovirus containing codon-modified HPV 16 L1 gene]. AB - OBJECTIVE: To construct recombinant adenovirus containing codon-modified HPV16L1 gene, and evaluate systemic and mucosal immunological responses induced after immunization with the recombinant virus. METHODS: The recombinant adenovirus rAd mod.HPV16L1 was constructed by Admax kit. The C57 BL/6 mice were immunized by purified rAd-mod.HPV16L1 through different inoculation routes. The immunological effect was evaluated by testing the specific neutralizing antibodies in sera and vaginal secretions of immunized mice through indirect ELISA and neutralization assay based HPV pseudovirus. RESULTS: The result showed that intramuscular immunization could induce good systemic immunity, but the mucosal immunity was too weak, and immunization via intranasal route could induce satisfactory immunity both in sera and vaginal secretions, while intravaginal immunization failed to induce any specific immunological responses either in sera or vaginal secretions. CONCLUSION: The recombinant adenovirus containing codon- modified HPV16L1 gene was successfully constructed. Immunization through intranasal route could induce satisfactory immunity both in sera and vaginal secretions, while intramuscular immunization could only induce high titer of neutralizing antibodies in sera. PMID- 18414688 TI - [Clinical characteristics of patients with hemorrhagic fever with renal syndrome]. AB - OBJECTIVE: Patients with typical clinical manifestations of Hemorrhagic fever with renal syndrome (HFRS) are becoming fewer. We conducted analysis on clinical features of HFRS in order to reduce the mistakes in diagnosis. METHODS: 64 patients were diagnosed as HFRS during May, 2000 to June, 2006 in our hospital. All the patients' serological tests (HFRS-NP-specific IgM, IgG antibody) by ELISA method were positive. We collected their clinical manifestations and test results. SPSS 12.0 was used in our statistical analysis. RESULTS: Among the 64 patients, 71.6% of all the cases occurred from Feb. to June. Most of patients were admitted to the hospital with untypical manifestation. Only 30.6% patients appeared headache, lumbago, and pain of orbital cavity. 32.8% patients had obviously signs of injection and hemorrhage. However, there were 90.6% patients with headache and 84.4% patients with nausea or vomit. Hypotensive or oliguric phases were absent in 56.3% patients. There were only 31.3% patients with all five stages. Thrombocytopenia (79.7%) and heavy proteinuria (71.9%) were common. But 54.7% of patients shown normal or even decreased white blood cell count. Only 2/3 of patients had elevated serum creatinine (Cr). Liver involved was common showing as elevated aminotransferase. ALT level was not always parallel to Cr level. There was an opposite trend between them. CONCLUSION: We must recognized the untypical manifestations of HFRS. Further study focus on pathogenesis was useful for diagnosis and therapy. PMID- 18414691 TI - [WU polyomavirus and KI polyomavirus detected in specimens from children with acute respiratory tract infection in China]. AB - OBJECTIVE: To investigate newly identified polyomavirus WUV and WUV and KIPyV are associated with acute respiratory infections in China, tests were developed to detect WUV and KIPyV gene fragments from nasopharyngeal aspirates collected from children with ARI fron Nov. 2006 to Oct. 2007. METHODS: A total of 318 clinical samples were tested for WUV and KIPyV using PCR method. The positive products were sequenced and compared with those in GenBank. RESULTS: 14 of the 318 Samples were positive (WUV was 2.2%, KIPyV was 2.2%). All of children who were positive for WUV or KIPyV had respiratory illness. CONCLUSION: Polyomavirus WU and KIPyV infection may be associated with upper and lower respiratory diseases. PMID- 18414692 TI - [Epidemiological survey on the infection of hepatitis E virus among pigs in Henan province]. AB - OBJECTIVE: To investigate hepatitis E virus (HEV) infection among pigs in Henan province. METHODS: A total of 623 swine sera, collected from 5 districts, were divided into two groups, under 3-month of age and over 3-month of age. They were tested for HEV antigen and antibody by using ELISAs, respectively. The sera positive for HEV antigen were tested for HEV RNA with RT-PCR. The positive products of RT-PCR were cloned and sequenced. RESULTS: The positive rates of anti HEV antibody of the groups under 3-month and over 3-month of age were 90.27% and 92.55%, respectively, without statistical difference, while those of HEV antigen were 15.93% and 5.69%, respectively, with significant difference. The positive rates of anti-HEV antibody and HEV antigen were significantly different among different districts. HEV RNA was detectable in 5 of 47 HEV antigen positive samples. The sequence analysis showed that in 4 of 5 specimens the sequence belonged to genotype 4 while in the remaining one the sequence was genotype 1. CONCLUSION: The prevalence rate of HEV infection in pigs was high in Henan province and the rate differed in different districts. PMID- 18414693 TI - [Analysis of the variation of 32 HIV-1 V3 loop from infected Chinese blood/plasma donors]. AB - OBJECTIVE: To characterize the variation in V3 loop of HIV-1 B'strains circulating in Chinese blood donors. METHODS: The c2-c3 regions of the HIV envelop gene were amplified by nest-PCR from 32 HIV-1-infected blood donors in He Nan province in China. The BIOEDIT and MEGA software are used to analyze the sequences of V3 loop. RESULTS: There are five types of central motifs of the 32 samples, in which GPGR and GPGQ are most common. More variations associated with T tropic/SI phenotype can be seen in AIDS group. CONCLUSION: The V3 tip motifs of HIV-1B' strains circulating in Chinese blood donors are various, the different characterization of V3 loop between AIDS and asymptomatic patients indicates different biological phenotype and pathogenesis which warrant additional investigation. PMID- 18414694 TI - [A serological survey of Epstein-Barr virus infection in children in Beijing]. AB - OBJECTIVE: To understand the prevalence of Epstein-Barr virus (EBV) infection in urban and rural areas of Beijing using the serological method. METHODS: Totally 589 serum samples were collected from children in Beijing urban and rural areas who were 0--14 years old and tested with Viron-Seron ELISA classic EBV virus capsid antigen IgG antibody (EBV VCA IgG) kit. Optical density of serum samples was obtained at the wavelength of 405 nanometers. Sero-positive or negative samples were determined according to standard curve and cut-off attached in ELISA classic EBV VCA IgG kits. The activity of EBV VCA IgG was calculated by using special formula. The percentage and activity of EBV VCA IgG from Beijing children were compared with SPSS 13.0 between the urban and rural areas. RESULTS: The percentage of EBV VCA IgG seropositive samples was 83.6%, and 80.8% in those from urban and 86.2% in those from rural areas. The peak value of EBV infection was 71% seen among children under the age of 3 years, and in urban area the rate was 67.7%, which was lower than that in the rural area (75.3%), and was 82.5% by the age of 6, which was lower than the data (up to 90%) reported 30 years ago. There was a significant difference in EBV infection rate and VCA IgG activities in children at different ages between urban and rural areas (P < 0.05). CONCLUSION: The rate of EBV infection in children living in urban area was lower by the age of 6 years. The primary infection of EBV occurred late in part of children lived in urban area. PMID- 18414695 TI - [Cloning of PD-1 gene and its prokaryotic expression in Escherichia coli]. AB - OBJECTIVE: To clone human PD-1 gene, construct a prokaryotic expression plasmid and express in E. coli. METHODS: The human PD-1 cDNA was cloned by RT-PCR from the total RNA, which was extracted from peripheral blood lymphocyte cell of the patient with chronic hepatitis B. Recombinant PD-1 protein was been expressed and purified after the prokaryotic expression plasmid had been constructed. It was identified by SDS-PAGE, DNA sequencing and amino acid sequencing. RESULTS: The PD 1 gene was cloned and confirmed by DNA sequencing. The recombinant protein was expressed in E. coli. The purified protein was obtained, then been confirmed by amino acid sequencing. CONCLUSION: The human PD-1 gene was successfully cloned and expressed in E. coli, which lays the foundation for further study on the function and application of PD-1. PMID- 18414696 TI - [Relationship between the genetic polymorphism of CYP3A5 gene and the clinical response to bicyclol in chronic hepatitis B]. AB - OBJECTIVE: To investigate the relationship between the genetic polymorphisms of CYP3A5 and the clinical effectiveness of Bicyclol on patients with chronic hepatitis B to make individual medication possible. METHODS: 34 cases of chronic hepatitis B were treated by bicyclol tablets for 24 weeks. Liver function indexes (ALT and AST) were determined before and after treatment. Blood CYP3A5 genotyping of each patient was determined by the PCR-RFLP analysis. RESULTS: All subjects were genotyped for the CYP3A5*3 gene and divided into different group. The groups comprised subjects with CYP3A5*3 carriers (n=18) and CYP3A5*1 carriers (n=16) which include CYP3A5*1/*1 (n=2) and CYP3A5*1/*3 (n=14). Compared with pre treatment, the serum ALT and AST levels were decreased obviously in all patients. The mean percentage reduction of serum ALT and AST levels were significantly greater in subjects with CYP3A5*3 carriers (79.73% and 74.76%) than in those with CYP3A5*1 carriers (65.90% and 49.63%; P < 0.05) The recovery rates of ALT and AST were significantly highter in CYP3A5*3 carriers than those in CYP3A5*1 carriers (P < 0.05). CONCLUSION: CYP3A5 genotype has an impact on the therapeutic effects of Bicyclol. The subjects with CYP3A5*3 carriers is more effective than the subjects with CYP3A5*1 carriers. CYP3A5 genotyping may be helpful in predicting therapeutic effects of Bicyclol especially in the terms of decreasing ALT and AST. PMID- 18414697 TI - [Enhanced protein production of Vif and APOBEC3G by HIV-1 Vpr]. AB - OBJECTIVE: Goal of this study was to test the potential regulatory effects of Vpr on Vif and Vif-mediated degradation of APOBEC3G. METHODS: The Vpr effect was first tested in a fission yeast RE007 strain that carries a single integrated copy of vpr gene in the chromosome and transformed with a vif-expressing plasmid. Similar tests were also carried out in a muristerone A vpr-inducing HEK293 mammalian cell line that were transfected with the plasmids expressing vif and/or APOBEC3G. Western Blot analyses were used to measure the corresponding protein levels under different experimental conditions. RESULTS: Expression of HIV-1 vpr appears to enhance the protein levels of Vif both in fission yeast and mammalian cells. A similar enhancement effect of APOBEC3G by Vpr was also detected in mammalian cells. Interestingly, however, the increased Vif protein level by Vpr did not result in more APOBEC3G degradation than without Vpr, indicating a potential regulatory effect of Vpr on Vif-mediated proteolysis of APOBEC3G. CONCLUSION: To our knowledge, this is the first report describing a potentially conserved and regulatory effect of HIV-1 Vpr on Vif and Vif-mediated protein degradation of APOBEC3G. PMID- 18414698 TI - [The genetic stability of non-replicating recombinant adenoviruses on 293 cells during continuous passage]. AB - OBJECTIVE: To investigate the genetic stability during the serial passages of non replicating recombinant adenoviruses based on the novel AdEasy system. METHODS: Four non-replicating adenoviruses expressing rotavirus antigens, which were generated by the AdEasy system, were used as models and continuously propagated on 293 cells for 20 passages. Samples of the infected cells were collected at every 5 passages for the PCR analysis of the inserted rotavirus genes, replication-competent adenoviruses (RCAs) as well as the Western blot examination of the expression of the certain rotavirus genes. RESULTS: The adenoviruses can be stably propagated on 293 cells. The existence and the stable expression of inserted rotavirus genes were able to be detected during the serial passage. The RCAs were not found within the first 10 passages, but the rvAdG2VP7(o) at passage 15 and the rvAdG2VP7(o) and rvAdG1VP7(o) at passage 20. CONCLUSION: The non replicating adenoviruses showed promising genetic stability during the continuous passage on 293 cells. The RCAs normally will not be detectable within 10 continuous passages. The results indicated the potential of such recombinant adenoviruses in the research and development of the gene therapies and adenoviral vectored vaccines. PMID- 18414699 TI - [The investigation of HEV epidemiology, inferior clinical infection and viral toxemia in blood donors in Quzhou area]. AB - OBJECTIVE: To investigate the infection of HEV in Quzhou area of Zhejiang Province. METHODS: All sera from blood donors in the central blood bank of Quzhou from April 2006 to April 2007 were used. Anti-HEV IgG and anti-HEV IgM were measured by EIA. RT-PCR was also performed to the samples with positive anti-HEV IgM. Genotype and sequence homology were analyzed after sequencing. RESULTS: The positive ratio of anti-HEV IgG was 40.60%, in which the male infection ratio was higher than the female significantly (43.09% VS 36.09%; chi2=22.6; P < 0.01). The infection ratio was increased with age. The positive ratio of anti-HEV IgM was 0.43%. The positive ratio of anti-HEV IgG and the titers of antibody were higher in the inferior clinical infectors with positive anti-HEV IgM than the negative ones (P < 0.05). Two samples were positive in HEV PCR among 21 samples with positive anti-HEV IgM. The toxemia ratio was 0.4% of all the donors. And the genotype of the two samples with toxemia were both HEV-IV. CONCLUSION: The HEV infection was correlation with age and sex significantly and the infection occurred in the adults mainly in Quzhou area. HEV toxemia was not infrequency in the blood donors. PMID- 18414700 TI - [Antigenic properties of mutant hepatitis B virus surface antigen]. AB - OBJECTIVE: To study the antigenic properties of mutant hepatitis B virus surface antigen, to understand the sensitivity of the commercially available HBsAg assays to the variants and to reduce the undetectability of the variants. METHODS: Recombinant eukaryotic expression plasmids for HBsAg. The recombinant eukaryotic expression plasmids pSS1adr, pSS1adw2, pSS1adw2- 145Arg, pSS1adr-126 Asn and pSS1adr-126Ser were transfected into COS-7 cells. HBsAg in the supernatants of transfected cells was detected by using different commercial ELISA kits. RESULTS: The absorbance value of pSS1adr-126 Asn and pSS1adr-126Ser plasmids were similar to that of the wild type HBsAg, the absorbance value of pSS1adw2-145Arg plasmids was lower than that of the wild type HBsAg. CONCLUSION: It is estimated that the antigenicity of HBsAg mainly depended on the amino acid sequence of "a" antigen determinant and its conformation, so 145 amino acid substitutions led to the change of conformation and the antigenicity of variant HBsAg was lower than that of the wild type. PMID- 18414701 TI - [The mechanism of HBV infection of human trophoblast cell]. AB - OBJECTIVE: To observe the changes of human trophoblast cells after infected with hepatitis B virus. METHODS: HBV positive serum was used to infect human trophoblast cells in vitro. HBsAg in cell culture medium were detected by ELISA method and HBV DNA in cell culture medium and cells were detected by PCR method. HBV fluorescence polymerase chain reaction diagnose kit were used to detect the HBV DNA concentration. Ultra structure of trophoblast cells were observed with transmission electron microscopy (TEM). RESULTS: HBsAg could be detected in infection group by ELISA. Infection group cell culture medium and infection group cells were HBV DNA positive. HBV DNA concentrations in HBV infection cell culture medium in 0, 12, 36, 60, 84 h after extensively PBS washed were < 10(3), 3 x 10(4), 6 x 10(5), 5 x 10(5), 3 x 10(5) copies/mL. HBV infected trophoblast cells were found many forms of endosomes, some of which contents virus like particle. CONCLUSION: HBV might take advantage of clathrin-mediated endocytosis to enter trophoblast cell, which might lead to cell infection or across the cell bar by transcytosis. PMID- 18414702 TI - [Efficacy of antiviral treatment on intrahepatic HBV DNA and histology in HBeAg positive chronic hepatitis B patients]. AB - OBJECTIVE: To evaluate the effect of antiviral agents on intrahepatic HBV DNA and histology in HBeAg-positive chronic hepatitis B patients. METHODS: Thirty-five patients were treated with lamivudine, 16 with interferon alfa (INF-alpha), 24 with sequential Lamivudine and INF-alpha. The total duration of therapy was 12 months. Intrahepatic HBV DNA was measured quantitatively by real-time polymerase chain reaction. RESULTS: There was significant change in all parameters of the groups of patients at the end of treatment (P < 0.05). The patients treated with sequential treatment had slightly higher HBeAg seroconversion rate (38.1%) than that of the other patients (P=0.1352). The baseline levels of intrahepatic HBV DNA in the patients with HBeAg seroconversion or undetectable serum HBV DNA were significantly lower than that of the other patients (P < 0.05). CONCLUSION: Antiviral agents could effectively inhibit intrahepatic HBV DNA and improve hepatic histology. The patients with low baseline intrahepatic HBV DNA level may achieve better antiviral efficacy. Sequential treatment might produce high HBeAg seroconversion rate. PMID- 18414703 TI - [Increased expression of inducible nitric oxide synthase in chronic hepatitis B patients is correlated with histopathological grading and staging]. AB - OBJECTIVE: To investigate the intrahepatic expression of inducible nitric oxide synthase (iNOS) in patients with chronic hepatitis B (CHB) and its relation to liver histopathology. METHODS: The intensity and distribution of the immunohistochemical staining of intrahepatic iNOS were studied in the liver biopsy specimens obtained from 74 patients with CHB and statistical analyses were performed between intrahepatic iNOS and ALT, HbeAg, HBV DNA grading of liver inflammation and staging of fibrosis. Seven histologically normal liver sections were used as a control group. RESULTS: Compared with the control group, the intrahepatic iNOS immunoexpression was significantly higher in patients with CHB (P < 0.05), iNOS immunoreactivity was observed mainly in hepatocytes showing a predominant cytoplasmic staining, with the positive liver cells distributed diffusely throughout the hepatic lobule. Immunopositive staining could also be detected in Kupffer cells, sinusoidal lining cells and vascular endothelial cells. Compared with patients with normal ALT, the hepatocellular iNOS immunoexpression was significantly higher in patients with elevated ALT (P < 0.05) and the iNOS immunoexpression was significantly correlated with the serum level of ALT (r=0.601, P=0.000). Statistical analysis also showed that the intrahepatic iNOS immunoexpression was positively correlated with the grading of liver inflammation and the staging of liver fibrosis (r=0.660, P=0.000; r=0.507, P=0.000). No significant correlation between iNOS and HBeAg and HBV DNA was detected. CONCLUSION The intrahepatic expression of iNOS is elevated in chronic hepatitis B patients and correlated well with the severity of the disease, which indicated that inducible nitric oxide synthase may have a critical role in the pathogenesis of chronic viral hepatitis B. PMID- 18414704 TI - [Acute transverse myelitis associated with coxsackievirus B---A retrospective analysis of 7 patients]. AB - OBJECTIVE: Acute Transverse myelitis (ATM) is a focal inflammatory disorder of the spinal cord, resulting in motor, sensory, and autonomic nerve dysfunction. There is often a clearly defined rostral border of sensory dysfunction. Nowadays, the pathogenesis of ATM is not clear. The present study aimed to understand possible relationship between ATM and infection with Coxsackievirus B. METHODS: IgM antibody against Coxsackievirus B was detected in cerebrospinal fluid of 33 patients with ATM. RESULTS: In 7 of the 33 cases with ATM, the IgM andtibody Coxsackievirus B (CVB) was positive. No infections with other pathogens were found at the onset of the disease. CONCLUSION: The pathogenesis of ATM may involve infection with Coxsackievirus B. PMID- 18414705 TI - [Study of the subcellular location of the human gene 6 transactivated by nonstructeral protein 5a of hepatitis C virus]. AB - OBJECTIVE: To found the subcellular location of the human gene 6 transactivated by nonstructural protein 5A of hepatitis C virus (NS5ATP6). METHODS: Green fluorescent protein (GFP) expression vector pEGFP- NS5ATP6 was established. The pEGFP- NS5ATP6 was transfected into HepG2 cells, and analyze the subcellular location of the proteins expressed by NS5ATP6 through Green fluorescent microscopy after 24 hours. RESULTS: The pEGFP- NS5ATP6 gene was successful cloned, NS5ATP6 can express protein in cells and subcellularly located in cell plasma. CONCLUSION: NS5ATP6 can express protein, and the protein expressed by NS5ATP6 subcellularly located in cell plasma. PMID- 18414706 TI - [Prokaryotic expression, purification of prM of JEV and preparation of monoclonal antibody]. AB - OBJECTIVE: To prepare monoclonal antibody (mAb) against prM epitope. METHODS: The gene encoding prM was isolated using RT-PCR from brain of JEV infected mouse and cloned into prokaryotic expression vector pET-32a. Recombinant plasmid was transformed into E.coli BL21/DE3/LysS, then the transformed cells were expressed with the induction of IPTG. The expression and purification of the prM protein was analyzed by SDS-PAGE. The BALB/c mice were immunized with purified prM protein. Hybridoma cell lines secreting monoclonal antibodies against prM were established after cell fusion of mouse splenic cell and P3-X63-Ag8.653 cells. The specificity of mAb was identified by ELISA, Western Blot and Immunohistochemistry assay. RESULTS: mAb against prM epitope of JEV was prepared successfully. CONCLUSION: The obtained prM specific mAb was valuable for the prevention and dignosis of Japanese encephalitis. PMID- 18414708 TI - Requiem for the term 'carcinoid tumour' in the gastrointestinal tract? AB - Use of the term 'carcinoid tumour' to describe a unique type of tumour in the gastroenteropancreatic system is endemic in the medical literature and in daily clinical and pathological parlance. However, it is a somewhat misleading moniker because a spectrum of histopathological changes and hence, biological outcomes may occur in these tumours. The World Health Organization classification scheme recommends the use of the terms neuroendocrine tumours or carcinomas, which may be stratified as well-differentiated neuroendocrine tumours with benign or uncertain behaviour, well-differentiated tumours with low-grade neuroendocrine carcinoma behaviour and high-grade neuroendocrine carcinomas. These categories may be applied within different sites in the gastrointestinal tract and pancreas, and convey a sense of biological behaviour. In addition, a recently suggested tumour-node-metastasis scheme has been proposed and awaits clinical validation and acceptance. Thus, the term 'carcinoid' has served its purpose well, but its use should be phased out in favour of 'neuroendocrine tumour' or 'neuroendocrine carcinoma'. PMID- 18414707 TI - Canadian credentialing guidelines for esophagogastroduodenoscopy. PMID- 18414709 TI - Publicly funded pegylated interferon-alpha treatment in British Columbia: disparities in treatment patterns for people with hepatitis C. AB - BACKGROUND: An estimated 60,000 British Columbians are chronically infected with the hepatitis C virus (HCV); 10% to 20% will develop cirrhosis after 20 years and 5% to 10% of these will develop hepatocellular carcinoma. Although treatment may prevent cirrhosis and liver cancer, and improve quality of life, availability is limited. METHODS: Individuals with HCV genotypes 1, 4, 5 and 6 who underwent baseline HCV-RNA tests between January 1, 2003 and December 31, 2005, and were eligible for publicly funded treatment through PharmaCare were linked to British Columbia's reportable disease database. Patterns in treatment were examined, including age at treatment, sex, location, time to treatment from HCV diagnosis and seasonality of treatment. RESULTS: When corrected for HCV prevalence, men were more likely to receive treatment than women (RR 1.16, 95% CI 1.02 to 1.31). Patients aged 35 to 54 years and 55 years or older were 3.45 times (95% CI 2.80 to 4.26 times) and 4.49 times (95% CI 3.55 to 5.69 times), respectively, more likely to initiate treatment than 15- to 34-year-olds. Differences were noted between health authorities. Patients in rural health service delivery areas (HSDAs) were 1.25 times (95% CI 1.10 to 1.42 times) more likely to receive treatment than those in urban HSDAs. Patients had an average lapse of four years between HCV diagnosis and receiving treatment. The highest proportion of patients initiated therapy between January and March (36.5%), with the lowest between October and December (less than 14%). CONCLUSIONS: This data linkage enabled us to identify populations less likely to receive publicly funded treatment. Rural HSDAs have higher rates of therapy initiation; this pattern merits further research but may be a result of integrated prevention and care projects in rural areas. Policy changes to the current PharmaCare funding co-payment schedules could reduce seasonal variability of treatment initiations throughout the year. PMID- 18414710 TI - High-resolution esophageal manometry: a time motion study. AB - INTRODUCTION: High-resolution manometry (HRM) of the esophagus is a new technique that provides a more precise assessment of esophageal motility than conventional techniques. Because HRM measures pressure events along the entire length of the esophagus simultaneously, clinical procedure time should be shorter because less catheter manipulation is required. According to manufacturer advertising, the new HRM system is more accurate and up to 50% faster than conventional methods. OBJECTIVE: To test the hypothesis that clinical testing with HRM requires less procedural time than a standard water perfusion (WP) method. METHODS: Forty-one consecutive patients were studied (20 underwent WP and 21 underwent HRM). Using time-motion analysis, the start and end times for each task associated with performing the study were recorded. Patient discomfort and study quality were also assessed by using five- and four-point qualitative scales, respectively. RESULTS: Total procedure time was reduced on average by 25.6% in the HRM group (from 41.8 minutes with WP to 30.7 minutes with HRM, P<0.05). There was no significant difference in the discomfort scores reported by the study subjects and no difference in study quality. CONCLUSIONS: HRM requires less time to complete than conventional manometry and should therefore shorten the wait-times of patients scheduled for esophageal manometry and have a significant impact on the cost of performing this commonly used clinical investigation. PMID- 18414711 TI - Barrett's esophagus and cardiac intestinal metaplasia: two conditions within the same spectrum. AB - BACKGROUND: Immunostaining for cytokeratin 7 (CK7) and cytokeratin 20 (CK20) has a characteristic pattern in Barrett's esophagus (BE), but reports regarding its sensitivity and specificity are inconsistent. Intestinal metaplasia of the gastric cardia (CIM) is histologically similar to BE, but with no abnormal endoscopic findings. OBJECTIVES: To evaluate the sensitivity and specificity of a semi-quantitative CK7/CK20 immunostaining pattern for the diagnosis of BE, and to further elucidate the pathogenesis of CIM. METHODS: Tissues were examined by hematoxylin and eosin and periodic acid schiff/alcian blue stains, and then were immunostained with CK7 and CK20 antibodies. Correlations with other clinical parameters were statistically analyzed. RESULTS: When values were revised based on follow-up data and auxiliary testing, all BE cases (100%) displayed the characteristic BE CK7/CK20 immunostaining pattern, compared with 66% of CIM cases. In the subgroup of patients who were endoscopically and immunohistochemistry-positive but histologically negative, all patients except for one had documented BE when clinical history, auxiliary testing and follow-up were evaluated. There were no statistically significant differences between BE and CIM regarding Helicobacter pylori infection or the type of metaplasia (complete versus incomplete). The sensitivity of the CK7/CK20 pattern reached 100% in the subgroup of CIM patients with a history of acid reflux. Of 26 cases of CIM where follow-up was available, four cases (15%) progressed to BE, and one developed dysplasia. All four cases showed the BE pattern of CK7/CK20 staining and were negative for H pylori infection. CONCLUSIONS: A semiquantitative CK7/CK20 pattern can be used to confirm BE even in the absence of histological evidence. The subgroup of CIM with acid reflux may develop into BE and may need closer follow-up. PMID- 18414712 TI - Evaluation of oral cannabinoid-containing medications for the management of interferon and ribavirin-induced anorexia, nausea and weight loss in patients treated for chronic hepatitis C virus. AB - OBJECTIVES: The systemic and cognitive side effects of hepatitis C virus (HCV) therapy may be incapacitating, necessitating dose reductions or abandonment of therapy. Oral cannabinoid-containing medications (OCs) ameliorate chemotherapy induced nausea and vomiting, as well as AIDS wasting syndrome. The efficacy of OCs in managing HCV treatment-related side effects is unknown. METHODS: All patients who initiated interferon-ribavirin therapy at The Ottawa Hospital Viral Hepatitis Clinic (Ottawa, Ontario) between August 2003 and January 2007 were identified using a computerized clinical database. The baseline characteristics of OC recipients were compared with those of nonrecipients. The treatment-related side effect response to OC was assessed by c2 analysis. The key therapeutic outcomes related to weight, interferon dose reduction and treatment outcomes were assessed by Student's t test and c2 analysis. RESULTS: Twenty-five of 191 patients (13%) initiated OC use. Recipients had similar characteristics to nonrecipients, aside from prior marijuana smoking history (24% versus 10%, respectively; P=0.04). The median time to OC initiation was seven weeks. The most common indications for initiation of OC were anorexia (72%) and nausea (32%). Sixty-four per cent of all patients who received OC experienced subjective improvement in symptoms. The median weight loss before OC initiation was 4.5 kg. A trend toward greater median weight loss was noted at week 4 in patients eventually initiating OC use (-1.4 kg), compared with those who did not (-1.0 kg). Weight loss stabilized one month after OC initiation (median 0.5 kg additional loss). Interferon dose reductions were rare and did not differ by OC use (8% of OC recipients versus 5% of nonrecipients). The proportions of patients completing a full course of HCV therapy and achieving a sustained virological response were greater in OC recipients. CONCLUSIONS: The present retrospective cohort analysis found that OC use is often effective in managing HCV treatment related symptoms that contribute to weight loss, and may stabilize weight decline once initiated. PMID- 18414713 TI - The burden of hepatitis C virus infection is growing: a Canadian population-based study of hospitalizations from 1994 to 2004. AB - BACKGROUND: Nearly 1% of Canadians are infected with the hepatitis C virus (HCV). Simulation analyses have suggested that HCV will place an increasing burden on the health care system as the infected population ages, but supportive clinical data are limited. OBJECTIVES: To study temporal trends in HCV-related hospitalizations and predictors of increased health care utilization from a Canadian population-based perspective. METHODS: An administrative hospitalization database from the Calgary Health Region was used to identify patients who were admitted for HCV between 1994 and 2004. The primary outcomes were liver-related HCV hospitalizations, length of stay, hospital costs and in-hospital mortality. Average annual growth rates in outcomes were calculated and subgroup analyses were conducted according to age, sex and HIV/HCV coinfection status. RESULTS: Between 1994 and 2004, there were 4002 HCV-related hospitalizations; 22% were liver-related. Liver-related hospitalizations, lengths of stay and in-hospital mortality increased approximately fourfold or an average of 15% to 18% annually (P<0.0005). Patients aged 40 to 59 years and HIV/HCV coinfected patients experienced the largest average annual growth rates (19% to 27% and 30% to 40%, respectively; P<0.0005), reflecting the accelerated natural history of HCV in these subgroups. Hospital costs for liver-related HCV hospitalizations increased by an average of 41% annually (P=0.001) between 2000 and 2004. The average annual increase in liver-related hospitalizations remained significant in a sensitivity analysis, even when 75% of HCV cases were under-reported in 1994. CONCLUSIONS: The present studies' findings confirm the growing burden of HCV on the Canadian health care system. Strategies to prevent HCV infection and maximize the dissemination and most effective use of potentially curative antiviral therapies are necessary to reduce these trends. PMID- 18414714 TI - The use of budesonide in the treatment of autoimmune hepatitis in Canada. AB - BACKGROUND: Autoimmune hepatitis (AIH) is a chronic inflammatory disease that is successfully treated with prednisone and/or azathioprine immunosuppressive therapy in 70% to 80% of patients. The remaining patients are intolerant or refractory to these standard medications. Budesonide, a synthetic glucocorticoid, undergoes a high degree of first-pass metabolism, reducing its systemic bioavailability, and has a 15-fold greater affinity for the glucocorticoid receptor than prednisolone. Budesonide may be a potentially useful systemic steroid-sparing immunosuppressive agent in the treatment of AIH. OBJECTIVE: To review the Canadian experience using budesonide to treat AIH. METHODS: Patients with AIH currently or previously treated with budesonide were identified through the Canadian Association for the Study of the Liver membership. Data were collected regarding their clinical and treatment history. RESULTS: A total of nine patients were identified. All patients were female, with an average age of 39 years (range 12 to 66 years). The indications for budesonide were adverse side effects of prednisone in two patients, noncompliance with prednisone and azathioprine in one patient and intolerance to azathioprine resulting in prednisone dependence in the remaining six patients. Patients were treated in doses ranging from 9 mg daily to 3 mg every other day for 24 weeks to eight years. Seven of nine patients had a complete response, defined as sustained normalization of the aminotransferase levels. The remaining two patients were classified as nonresponders (less than a 50% reduction in pretreatment aminotransferase levels). CONCLUSIONS: In Canada, budesonide has been successfully used in seven of nine patients with autoimmune hepatitis who were either intolerant to prednisone and azathioprine or prednisone-dependent. No adverse effects were reported with budesonide. Budesonide is potentially a valuable treatment option for AIH patients refractory or intolerant to standard therapy, and is deserving of further study. PMID- 18414715 TI - Long-term survivorship of esophageal cancer patients treated with radical intent. AB - To investigate the recent trends in definitive management of esophageal cancer, the records of 138 consecutive patients treated with radical intent in a single institution between 1995 and 2003 were reviewed and analyzed. The median follow up period was 5.7 years (range 1.1 to 10.4 years). Seventy-seven patients were treated with radiation therapy (RT) only and 61 with combined regimens (CRT), in which RT was combined with either radical surgery or chemotherapy, or both. The overall survival of the entire cohort was 32% over two years and 20% over five years. The survivorship in the RT group was 17% over two years and 5% over five years. In the CRT group, 51% and 35% survived over two and five years, respectively. From all the potential prognostic factors examined by univariate and multivariate analyses, only male sex and use of CRT were strongly associated with better survivorship. There was no significant difference in the outcomes among the different regimens of CRT. Survivorship was not affected by the location or histology of the tumour, clinical stage, dose of RT or use of endoluminal brachytherapy in addition to external beam RT. There was a greater tendency to use RT only more often in older patients, but patient age did not affect survivorship. The proportion of patients treated with CRT did not change significantly over the last versus the first four years of the observed period. Combined regimens are undoubtedly superior to RT as a single modality. The long term survivorship of patients in a subgroup of our patients treated with combined modality protocols compared favourably with the previously reported results in the literature and specifically in prospective randomized trials. However, the optimal combined modality regimen is yet to be defined. PMID- 18414716 TI - Does blood urea nitrogen level predict severity and high-risk endoscopic lesions in patients with nonvariceal upper gastrointestinal bleeding? AB - BACKGROUND: Nonvariceal upper gastrointestinal bleeding (UGIB) is a serious medical condition requiring prompt resuscitation and early endoscopic therapy in those with high-risk endoscopic lesions (HRLs). There are little or no data correlating sole blood urea nitrogen (BUN) level with the severity of nonvariceal UGIB or the presence of HRLs in the adult population. OBJECTIVES: To determine if the BUN level on presentation correlates with parameters of severity of UGIB (need for blood transfusion or intensive care unit [ICU] admission) or to the subsequent finding of HRL, and in so doing identify patients who will require early endoscopic intervention. METHODS: The Canadian Registry of patients with Upper Gastrointestinal Bleeding undergoing Endoscopy was used to identify patients enrolled from the McGill University Health Centre (Montreal, Quebec) who presented with or developed acute nonvariceal UGIB while admitted. All comparisons were performed using Student's t test or Wilcoxon's signed rank test, as appropriate. Logistic regression modelling using a stepwise method was performed to identify independent predictors of severe nonvariceal UGIB and HRL. RESULTS: Two hundred nine patients were enrolled in the study. The mean age was 67+/-18 years and 59.8% were male. The mean BUN level was 13.4+/-9.4 mmol/L. Univariate analysis demonstrated that the BUN level was a significant predictor of ICU admission (BUN 14.7+/-10.4 mmol/L versus 12.0+/-8.0 mmol/L, P=0.035). However, when adjusted for systolic blood pressure, BUN level became a weaker predictor of ICU admission, just failing to achieve statistical significance (OR 1.03, 95% CI 1.00 to 1.06; P=0.08). Univariate analysis also demonstrated that BUN level was not a statistically significant predictor of blood transfusion requirement (BUN 14.1+/-10.6 mmol/L versus 13.6+/-8.6 mmol/L, P=0.508), nor of HRL (BUN 14.2+/-10.7 mmol/L versus 12.9+/-8.6 mmol/L, P=0.605). CONCLUSION: In patients with nonvariceal UGIB, the BUN level at initial presentation is a weak predictor of the severity of UGIB as defined by ICU admission, but is not helpful in identifying patients with a HRL. PMID- 18414718 TI - Pneumoscrotum after colonoscopy. AB - Pneumoscrotum is an unusual problem that is very rarely associated with gastrointestinal endoscopy procedures. It has been reported to occur after colonoscopy and polypectomy. The present paper describes the case of an 81-year old man with benign pneumoscrotum that formed after polypectomy at the site of a previous rectal polyp. The pneumoscrotum was managed with conservative treatment. PMID- 18414717 TI - Endoscopic ultrasound-guided intratumoural therapy for pancreatic cancer. AB - Pancreatic cancer is the second most frequent gastrointestinal malignancy and carries a dismal prognosis. The current standard of care includes resection, if possible, as well as systemic chemoradiation therapy. Endoscopic ultrasound (EUS) is an established technique for the diagnosis and staging of pancreatic adenocarcinoma. Interventional EUS via fine needle injection (FNI) for the treatment of pancreatic cancer is a rapidly expanding field. The present article reviews the up-to-date developments in EUS FNI for intratumoural pancreatic cancer therapy, including antitumoural agents, immunotherapy, ablative techniques and new delivery systems. The therapeutic modalities discussed are currently under development and will hopefully reach clinical practice if benefit is demonstrated through clinical trials. EUS FNI may be an exciting new technique for the delivery of desperately needed novel therapies for pancreatic cancer. PMID- 18414719 TI - Development of bioinspired artificial photosynthetic systems. AB - The specific objective of this review is to describe recent development of bioinspired artificial photosynthetic systems and their applications. First, multi-step electron-transfer systems composed of electron donor-acceptor ensembles are presented, mimicking functions of the photosynthetic reaction center. However, a significant amount of energy is lost during the multi-step electron-transfer processes. Then, as an alternative to conventional charge separation functional molecular models based on multi-step long-range electron transfer within redox cascades, simple electron donor-acceptor dyads linked by covalent or non-covalent bonding have been developed to attain a long-lived and high-energy charge-separated state without significant loss of excitation energy. Such simple molecular dyads, capable of fast charge separation but extremely slow charge recombination, have significant advantages with regard to synthetic feasibility, providing a variety of applications including construction of organic solar cells and development of efficient photocatalytic systems for the solar energy conversion. PMID- 18414720 TI - Molecular electrostatic potentials of divalent carbon(0) compounds. AB - The molecular electrostatic potentials of divalent carbon(0) and divalent carbon(ii) compounds are calculated and the results are compared with theoretically predicted proton affinities and complexation energies with BH(3). PMID- 18414721 TI - Efficient modification of Cu electrode with nanometer-sized copper tetracyanoquinodimethane for high performance organic field-effect transistors. AB - We report high performance organic field-effect transistors (OFETs) with the modified Cu bottom-contact electrodes. Efficient modification of the Cu electrodes with nanometer-size copper tetracyanoquinodimethane (Cu-TCNQ) increases the electrode/organic layer contact area and reduces contact resistance. We investigated the effect of the Cu-TCNQ morphology on the device performance. The pentacene-based OFETs with the modified Cu bottom-contact electrodes exhibited high device performance. The field-effect mobility up to 0.31 cm(2)/V s was achieved. To the best of our knowledge, this is the highest device performance for the OFETs with the bottom Cu electrodes ever reported. Consequently, our results provide an effective approach to fabricate high performance and low-cost OFETs. PMID- 18414722 TI - Performance of multi-configurational calculations for a 1,4 bis(phenylethynyl)benzene derivative conjugated molecule. AB - The theoretical challenge of finding a single method that quantitatively reproduces both the experimental low-lying excitation energies and the torsional barrier of a prototypical conjugated molecule, which could act as a molecular wire, has been addressed here. The results indicate that this goal can be reasonably achieved when multi-reference perturbation theory up to second order (MRMP2) based on a complete active space self-consistent field (CASSCF) wave function using large active spaces is used. The results obtained were also used to compare with less expensive Kohn-Sham (KS) density functional theory (DFT) calculations when applied to these properties. The results obtained with BLYP and B3LYP exchange-correlation functionals indicate that quantitative agreement with all the experimental data cannot be obtained with this methodology, with a clear dependence on the exchange-correlation form selected. We thus encourage a careful testing of pure and hybrid density functionals whenever KS DFT is used for the rational design of conjugated materials for charge conduits. PMID- 18414723 TI - Sandwich-like compounds based on bare all-boron cluster B(6)(2-). AB - Here, we theoretically predict antiaromatic double-decked compounds [DMB(6)](q-) (D = B(6)(2-), Cp(-); M = Li, Na, K, Be, Mg, Ca) as well as the triple-decked sandwich-like species. Being energetically higher than the fusion isomers, the homo-decked assembly species [B(6)MB(6)](q-) without and with counterions are less likely to be observed experimentally. The hetero-decked sandwich species are low-lying minima containing double-fold antiaromatic B(6)(2-) building blocks. Additionally, the well-retained double antiaromaticity is mainly ascribed to the ionic electrostatic interaction and the protection of rigid Cp-deck in order to avoid the fusion of B(6)(2-). Our results represent the first example that the antiaromaticity is well retained in assembled compounds as in the free B(6)(2-) cluster. Realization of the double antiaromatic B(6)(2-)-incorporated assembled compound is very promising. PMID- 18414724 TI - Theoretical study of multiphoton ionization of cyclohexadienes and unimolecular decomposition of their mono- and dications. AB - Quantum chemical calculations of the geometric structure, vertical excitation energies, and ionization potentials for the isomeric pair of 1,3- and 1,4 cyclohexadienes and their mono- and dications have been performed employing a variety of theoretical methods and basis sets. The computed ionization potentials and electronic excitation energies are used to evaluate the range of internal energies available for fragmentation of the cations following multiphoton resonance ionization of the cyclohexadienes in intense laser field. The conditions governing the competition between multiple ionization and decomposition of the ions are also discussed. Calculations of stationary points on the potential energy surfaces for various fragmentation channels and relative product yields at different available internal energies are then utilized to analyze the trends in branching ratios of major dissociation products of the 1,4 cyclohexadiene(2+) dication, which include C(3)H(3)(+) + C(3)H(5)(+), C(2)H(3)(+) + C(4)H(5)(+), and C(4)H(3)(+) + C(2)H(5)(+). PMID- 18414725 TI - Water interaction with hydrophobic and hydrophilic soot particles. AB - The interaction of water with laboratory soots possessing a range of properties relevant for atmospheric studies is examined by two complementary methods: gravimetrical measurement of water uptake coupled with chemical composition and porosity analysis and HTDMA (humidified tandem differential mobility analyzer) inference of water uptake accompanied by separate TEM (transmission electron microscopy) analysis of single particles. The first method clarifies the mechanism of water uptake for bulk soot and allows the classification of soot with respect to its hygroscopicity. The second method highlights the dependence of the soot aerosol growth factor on relative humidity (RH) for quasi monodisperse particles. Hydrophobic and hydrophilic soot are qualitatively defined by their water uptake and surface polarity: laboratory soot particles are thus classified from very hydrophobic to very hydrophilic. Thermal soot particles produced from natural gas combustion are classified as hydrophobic with a surface of low polarity since water is found to cover only half of the surface. Graphitized thermal soot particles are proposed for comparison as extremely hydrophobic and of very low surface polarity. Soot particles produced from laboratory flame of TC1 aviation kerosene are less hydrophobic, with their entire surface being available for statistical monolayer water coverage at RH approximately 10%. Porosity measurements suggest that, initially, much of this surface water resides within micropores. Consequently, the growth factor increase of these particles to 1.07 at RH > 80% is attributed to irreversible swelling that accompanies water uptake. Hysteresis of adsorption/desorption cycles strongly supports this conclusion. In contrast, aircraft engine soot, produced from burning TC1 kerosene in a gas turbine engine combustor, has an extremely hydrophilic surface of high polarity. Due to the presence of water soluble organic and inorganic material it can be covered by many water layers even below water saturation conditions. This soot demonstrates a gradual diameter growth factor (D(wet)/D(dry)) increase up to 1.22 at 93% relative humidity, most likely due to the presence of single particles with water soluble material heterogeneously distributed over their surface. PMID- 18414726 TI - Interaction of formic and acetic acid with ice surfaces between 187 and 227 K. Investigation of single species- and competitive adsorption. AB - The physical adsorption of formic (HC(O)OH) and acetic (CH(3)C(O)OH) acid on ice was measured as a function of concentration and temperature. At low concentrations, the gas-ice interaction could be analysed by applying Langmuir adsorption isotherms to determine temperature dependent partition constants, K(Lang). Using temperature independent saturation coverages (N(max)) of (2.2 +/- 0.5) x 10(14) molecule cm(-2) and (2.4 +/- 0.6) x 10(14) molecule cm(-2) for HC(O)OH and CH(3)C(O)OH, respectively, we derive K(Lang)(HC(O)OH) = 1.54 x 10( 24) exp (6150/T) and K(Lang)(CH(3)C(O)OH) = 6.55 x 10(-25) exp (6610/T) cm(3) molecule(-1). Via a van't Hoff analysis, adsorption enthalpies were obtained for HC(O)OH and CH(3)C(O)OH. Experiments in which both acids or HC(O)OH and methanol interacted with the ice surface simultaneously were adequately described by competitive adsorption kinetics. The results are compared to previous measurements and used to calculate the equilibrium partitioning of these trace gases to ice surfaces under conditions relevant to the atmosphere. PMID- 18414727 TI - Using (18)O/(16)O exchange to probe an equilibrium space-charge layer at the surface of a crystalline oxide: method and application. AB - The use of an (18)O/(16)O exchange experiment as a means for probing surface space-charge layers in oxides is examined theoretically and experimentally. On the basis of a theoretical treatment, isotope penetration profiles are calculated for (18)O/(16)O exchange across a gas-solid interface and subsequent diffusion of the labelled isotope through an equilibrium space-charge layer depleted of mobile oxygen vacancies and into a homogeneous bulk phase. Profiles calculated for a range of conditions all have a characteristic shape: a sharp drop in isotope fraction close to the surface followed by a normal bulk diffusion profile. Experimental (18)O profiles in an exchanged (001) oriented single crystal of Fe doped SrTiO(3) were measured by time-of-flight secondary ion mass spectrometry (ToF-SIMS). By extracting the space-charge potential from such profiles, we demonstrate that this method allows the spatially resolved characterization of space-charge layers at the surfaces of crystalline oxides under thermodynamically well-defined conditions. PMID- 18414728 TI - Design of equidistant and revert type precipitation patterns in reaction diffusion systems. AB - In the past years considerable attention has been devoted to designing and controlling patterns at the microscale using bottom-up self-assembling techniques. The precipitation process proved itself to be a good candidate for building complex structures. Therefore, the techniques and ideas to control the precipitation processes in space and in time play an important role. We present here a simple and technologically applicable technique to produce arbitrarily shaped precipitation (Liesegang) patterns. The precipitation process is modelled using a sol coagulation model, in which the precipitation occurs if the concentration of the intermediate species (sol) produced from the initially separated reactants (inner and outer electrolytes) reaches the coagulation threshold. Spatial and/or temporal variation of this threshold can result in equidistant and revert (inverse) type patterns in contrast to regular precipitation patterns, where during the pattern formation a constant coagulation threshold is supposed and applied in the simulations. In real systems, this threshold value may be controlled by parameters which directly affect it (e.g. temperature, light intensity or ionic strength). PMID- 18414729 TI - Study of the photoelectron and electron momentum spectra of cyclopentene using benchmark Dyson orbital theories. AB - A complete study of the valence electronic structure and related electronic excitation properties of cyclopentene in its C(s) ground state geometry is presented. Ionization spectra obtained from this compound by means of photoelectron spectroscopy (He I and He II) and electron momentum spectroscopy have been analyzed in details up to electron binding energies of 30 eV using one particle Green's function (1p-GF) theory along with the outer-valence (OVGF) and the third-order algebraic diagrammatic construction [ADC(3)] schemes. The employed geometries derive from DFT/B3LYP calculations in conjunction with the aug-cc-pVTZ basis set, and closely approach the structures inferred from experiments employing microwave spectroscopy or electron diffraction in the gas phase. The 1p-GF/ADC(3) calculations indicate that the orbital picture of ionization breaks down at electron binding energies larger than approximately 17 eV in the inner-valence region, and that the outer-valence 7a' orbital is also subject to a significant dispersion of the ionization intensity over shake-up states. This study confirms further the rule that OVGF pole strengths smaller than 0.85 foretell a breakdown of the orbital picture of ionization at the ADC(3) level. Spherically averaged (e, 2e) electron momentum distributions at an electron impact energy of 1200 eV that were experimentally inferred from an angular analysis of EMS intensities have been interpreted by comparison with accurate simulations employing ADC(3) Dyson orbitals. Very significant discrepancies were observed with momentum distributions obtained from several outer-valence ionization bands using standard Kohn-Sham orbitals. PMID- 18414730 TI - Binuclear Robson type Ni(ii) complex as a reactant supplementing our knowledge of the orientation effects in electrochemical kinetics. AB - The multistep reduction of a binuclear Ni(ii) Robson-type complex with a multidentate template-like organic ligand (formed from 4-tert-butyl-2,6 diformylphenol and 1,3-diaminopropane), Ni(2)L, is studied using the electron photoemission technique. The number of transferred electrons corresponding to a single reduction wave is found to be 8 per complex species. This value is attributed to both complete Ni(ii) reduction (with Ni metal formation) and ligand reduction. Contributions of Ni(ii) and ligand to acceptor orbital were estimated. Three initial subsequent steps correspond to electron transfer to mixed metal ligand orbital with comparable contributions. For more deep reduction, ligand contribution predominates. The first single-electron step is evidenced to be rate determining, with the rate constant of 0.03 cm(2) s(-1). The latter value is discussed in the framework of a semiquantitative analysis of the rate constants estimated in the framework of quantum-mechanical electron transfer theory for different orientations of Ni(2)L in the reaction layer. The analysis includes estimations of key kinetic parameters (electronic transmission coefficient, solvent- and intramolecular contributions to the total reorganization energy) which strongly rest on the results of quantum chemical modeling. The transmission coefficients at realistic electrode-reactant distances of the closest approach are below 0.001. This means that despite of the noticeable delocalization of Ni(2)L acceptor orbital, the electron transfer is diabatic. Predominating contribution to reorganization energy results from solvent and does not exceed 0.5 eV for any reactant orientation. The highest reactivity is predicted for a planar orientation located mostly outside the compact part of electric double layer. The Ni(2)L adsorption in planar and vertical orientations on mercury is addressed as well. The results give a clear explanation of the previously observed self-inhibition of "dark" reduction of Ni(2)L on mercury and independent data on the adsorption of these species. The discovered combination of various orientation effects is compared with effects observed for other reactants. PMID- 18414731 TI - Self-assembly and heterogeneous electron transfer properties of metallo octacarboxyphthalocyanine complexes on gold electrode. AB - Electrochemical properties of redox-active self-assembled molecular films of novel metallo-octacarboxyphthalocyanine (MOCPc, M = Fe, Co and Mn) complexes integrated with cysteamine (Cys) monolayer on gold electrodes via amide bonds were investigated. X-Ray photoelectron spectroscopy confirmed the appearance of the various elements in their expected chemical environment upon immobilization of these species. The heterogeneous electron transfer properties of the Au-Cys MOCPc molecular films using an outer-sphere ([Fe(CN)(6)](4-)/[Fe(CN)(6)](3-)) redox probe were studied using cyclic voltammetry and electrochemical impedance spectroscopy. The electron transfer rate constant (k(app)) depends markedly on the central metal of the metallophthalocyanine cores (k(app): Co > Mn > Fe). A strong pH dependence of the electron transport of the Au-Cys-MOCPc molecular films was found. The surface pK(a) values of the MOCPc complexes were essentially the same (ca. 7.5). The differences in the electron transports and ionization constants are discussed. The electrodes are sensitive to the electrooxidation of epinephrine in physiological pH conditions, peak potential (E(p)/V vs. Ag|AgCl, saturated KCl) decreasing as FeOCPc (0.20 V) < MnOCPc (0.26 V) < CoOCPc (0.34 V). PMID- 18414732 TI - Dissociative electron transfer to organic chlorides: electrocatalysis at metal cathodes. AB - The reductive cleavage of a series of organic chlorides, including chloroaromatics, benzyl chlorides, activated chloroalkanes and polychloromethanes, was investigated at Ag, Cu, Pd and glassy carbon (GC) electrodes in CH(3)CN + 0.1 M (C(2)H(5))(4)NClO(4). The silver cathode was either a 2-mm diameter disc, fabricated from Ag wire, or nanoclusters of average diameter d = 304 nm, prepared by electrodeposition on GC. Ag, Cu and Pd electrodes have shown remarkable electrocatalytic properties for the reduction of several compounds. The peak potentials recorded at these electrodes, for example, at upsilon = 0.1 V s(-1) are positively shifted by 0.3-0.8 V with respect to the reduction potentials measured at a non catalytic electrode such as GC. Electrocatalysis is strictly related to the concerted nature of the dissociative electron transfer to the carbon-chlorine bond. No catalysis is observed when the dissociative electron transfer to RCl occurs according to a stepwise mechanism involving the intermediate formation of a radical anion. The catalytic surfaces affect the reaction scheme, offering a more favourable route possibly through the formation of strongly adsorbed activated complexes. PMID- 18414733 TI - The genome BLASTatlas-a GeneWiz extension for visualization of whole-genome homology. AB - The development of fast and inexpensive methods for sequencing bacterial genomes has led to a wealth of data, often with many genomes being sequenced of the same species or closely related organisms. Thus, there is a need for visualization methods that will allow easy comparison of many sequenced genomes to a defined reference strain. The BLASTatlas is one such tool that is useful for mapping and visualizing whole genome homology of genes and proteins within a reference strain compared to other strains or species of one or more prokaryotic organisms. We provide examples of BLASTatlases, including the Clostridium tetani plasmid p88, where homologues for toxin genes can be easily visualized in other sequenced Clostridium genomes, and for a Clostridium botulinum genome, compared to 14 other Clostridium genomes. DNA structural information is also included in the atlas to visualize the DNA chromosomal context of regions. Additional information can be added to these plots, and as an example we have added circles showing the probability of the DNA helix opening up under superhelical tension. The tool is SOAP compliant and WSDL (web services description language) files are located on our website: (http://www.cbs.dtu.dk/ws/BLASTatlas), where programming examples are available in Perl. By providing an interoperable method to carry out whole genome visualization of homology, this service offers bioinformaticians as well as biologists an easy-to-adopt workflow that can be directly called from the programming language of the user, hence enabling automation of repeated tasks. This tool can be relevant in many pangenomic as well as in metagenomic studies, by giving a quick overview of clusters of insertion sites, genomic islands and overall homology between a reference sequence and a data set. PMID- 18414734 TI - A primer on ankyrin repeat function in TRP channels and beyond. AB - Transient receptor potential (TRP) channels are rapidly gaining attention as important receptors and transducers of diverse sensory and environmental cues. Recent progress in the field has provided new insights into the structure and function of the ankyrin repeat motifs present in the N-terminal cytosolic domain of many TRP channels. The topics addressed in this Highlight include the structural features of canonical ankyrin repeats, new clues into the functions these repeats perform in cells, and how this information can be applied to develop further experiments on TRP channels and other proteins containing ankyrin repeats. PMID- 18414735 TI - Signal transduction pathways used by NLR-type innate immune receptors. AB - Proteins from the nucleotide-binding domain, LRR containing (NLR) family are involved in sensing bacterial invasion and danger signals in mammalian cells. Activation of these molecules leads to inflammatory responses which help clearance of invading pathogens. Recent data now shed light on the signal transduction pathways used by NLR proteins. This review summarizes advances in our understanding of signalling through NLRs with special emphasis on the Nod1 and Nod2 pathways. PMID- 18414736 TI - How to tailor non-ribosomal peptide products--new clues about the structures and mechanisms of modifying enzymes. AB - Non-ribosomal peptide products often contain modified building blocks or post assembly line alterations of their peptide scaffolds with some of them being crucial for biological activity. These reactions such as halogenation, hydroxylation or glycosylation are mostly catalyzed by individual enzymes associated with the respective biosynthesis cluster. The versatile nature of these chemical modifications gives rise to a high degree of structural and functional diversity. Recent progress in this area enhances our insight about the mechanisms of these enzymes. Biotechnological applications might include the synthesis of novel, non-ribosomal peptide products or modified amino acid building blocks for pharmaceutical research. PMID- 18414738 TI - The nucleobase-ascorbate transporter (NAT) family: genomics, evolution, structure function relationships and physiological role. AB - This review summarizes knowledge concerning a ubiquitous plasma transmembrane protein family that mediates nucleobase or ascorbate secondary active transport (NAT). We show that prototype bacterial and mostly fungal members have become unique model systems to unravel structure-function relationships and regulation of expression, using classical and reverse genetics, as well as biochemical approaches. We discuss the importance of NAT-mediated ascorbate transport in mammals and how changes in substrate specificity, from different nucleobases to ascorbate, might have evolved at the molecular level. Finally, we also discuss how modelling NAT-purine interactions might constitute a step towards the use of NAT proteins as specific gateways for targeting pathogenic microbes. PMID- 18414737 TI - The importance of surfaces in single-molecule bioscience. AB - The last ten years have witnessed an explosion of new techniques that can be used to probe the dynamic behavior of individual biological molecules, leading to discoveries that would not have been possible with more traditional biochemical methods. A common feature among these single-molecule approaches is the need for the biological molecules to be anchored to a solid support surface. This must be done under conditions that minimize nonspecific adsorption without compromising the biological integrity of the sample. In this review we highlight why surface attachments are a critical aspect of many single-molecule studies and we discuss current methods for anchoring biomolecules. Finally, we provide a detailed description of a new method developed by our laboratory for anchoring and organizing hundreds of individual DNA molecules on a surface, allowing "high throughput" studies of protein-DNA interactions at the single-molecule level. PMID- 18414740 TI - A further contribution to the extreme variability of quadruplex structures from oligodeoxyribonucleotides containing inversion of polarity sites in the G-tract. AB - Structural insight into DNA quadruplex structures formed by oligodeoxyribonucleotides 3'TG5'-5'GGGT3' (QS55) and 5'TG3'-3'GGGT5' (QS33) is presented. NMR analysis reveals that QS33 forms a parallel-like four-fold symmetric quadruplex, while QS55 possesses a two-fold symmetry and is characterized by a tetrameric antiparallel quadruplex embedded between two parallel tracts. The results reported here describe unprecedented quadruplex complexes provided by peculiar structural features never reported to date. These structures might inspire the design of new aptameric nucleic acids characterized by novel structural motifs hardly realizable with unmodified DNA/RNA. PMID- 18414739 TI - Dynamic characterisation of the netrin-like domain of human type 1 procollagen C proteinase enhancer and comparison to the N-terminal domain of tissue inhibitor of metalloproteinases (TIMP). AB - The backbone mobility of the C-terminal domain of procollagen C-proteinase enhancer (NTR PCOLCE1), part of a connective tissue glycoprotein, was determined using 15N NMR spectroscopy. NTR PCOLCE1 has been shown to be a netrin-like domain and adopts an OB-fold such as that found in the N-terminal domain of tissue inhibitors of metalloproteinases-1 (N-TIMP-1), N-TIMP-2, the laminin-binding domain of agrin and the C-terminal domain of complement protein C5. NMR relaxation dynamics of NTR PCOLCE1 highlight conformational flexibility in the N terminus, strand A and the proximal CD loop. This region in N-TIMP is known to be essential for inhibitory activity against the matrix metalloproteinases and suggests that this region is of equal importance for NTR PCOLCE1, although the specific functional activity of the NTR PCOLCE1 domain is still unknown. Dynamics observed within the structural core of NTR PCOLCE1 that are not observed in N TIMP molecules suggest that although the two domains have a similar architecture, the NTR PCOLCE1 domain will show different thermodynamic properties on binding and hence the target molecule could be somewhat different from that observed for the TIMPs. ModelFree order parameters show that NTR PCOLCE1 has more flexibility than both N-TIMP-1 and N-TIMP-2. PMID- 18414741 TI - Photoinduced RNA interference using DMNPE-caged 2'-deoxy-2'-fluoro substituted nucleic acids in vitro and in vivo. AB - Various chemical modifications to RNA have been incorporated in attempts to improve their pharmacological properties for RNAi interference (RNAi). Recent studies have shown that small interfering RNA (siRNA) containing 2'-fluoro modifications can elicit gene silencing through RNAi. Despite developments in using chemical modifications for increased stability, safety, and efficiency of these therapeutics, they still face challenges of spatial and temporal targeting. One potential targeting strategy is to use photocaging techniques, which involve the covalent attachment of photolabile compounds to the effector nucleic acid species that block bioactivity until exposed to near UV light. In this study we demonstrate that fully 2'-fluorinated nucleic acids (FNAs) can be caged for photoactivated gene silencing in cell culture and in zebrafish embryos. This strategy combines the improvement in chemical and enzymatic stability associated with 2'-substitutions with the targeting ability of a photoinducible trigger. Statistical alkylation of FNAs with 1-(4,5-dimethoxy-2-nitrophenyl)diazoethane (DMNPE) improved resistance to enzymatic degradation, reduced RNAi effectiveness, and protected the biological system from toxic doses of the effector. Photo exposure to 365 nm light partially restored the silencing activity of the 2' fluoro siRNAs. These results suggest that photocaging may offer control over RNAi therapeutics for spatially and temporally directed activation, while improving enzymatic stability and potentially enabling therapeutic dosing via light dose intensity. PMID- 18414742 TI - Immobilization of monomeric organometallic molybdenum oxo and carbonyl complexes and their application in epoxidation reactions. AB - Various oxomolybdenum complexes are efficient and selective homogeneous catalysts in the presence of organic hydroperoxides. The increasing interest in the heterogenisation of such catalysts has led to an increased interest in finding suitable support materials for the complexes. These supports include ionic liquids, mesoporous materials and zeolites to name a few. Several methods are used to anchor the catalyst depending on the nature of the carrier material. The supported catalysts combine properties of homogeneous catalysts such as reactivity, control and selectivity with those of heterogeneous catalysts such as enhanced stability and recyclability. PMID- 18414743 TI - Synthesis and characterization of the first metallacyclooctane. AB - Ring closing metathesis (RCM) using Grubbs' 1st generation catalyst with cis [L(2)Pt(CH(2)CH(2)CH[double bond, length as m dash]CH(2))(CH(2)CH(2)CH(2)CH[double bond, length as m-dash]CH(2))] yields a platinacyclooctene which can be hydrogenated to the platinacyclooctane, L(2)Pt(CH(2))(7) [L(2) = Ph(2)PCH(2)CH(2)CH(2)PPh(2) (dppp) or Ph(2)PCH(2)CH(2)PPh(2) (dppe)] in high yield. PMID- 18414744 TI - Formation of acylruthenium promoted by coordination of AlMe(3) to (eta(4) cyclopentadienone)Ru(CO)(3). AB - The reaction of AlMe(3) with (eta(4)-tetraphenylcyclopentadienone)Ru(CO)(3) leads to rapid and quantitative formation of an adduct arising from coordination of the enone oxygen to aluminium, which undergoes alkylation at the Ru(CO)(3) moiety to give (eta(5)-C(4)Ph(4)C(OAlMe(2)))Ru(CO)(2)(COMe) concomitant with a change of hapticity of the dienone ligand. PMID- 18414745 TI - Ligands that enforce unnatural stereospinomers. AB - Computational evidence is provided for the existence of combinations of a stereoisomer and a spin state not previously observed in d(4) and d(8) complexes, achieved by means of encapsulating ligands that enforce a tetrahedral coordination. PMID- 18414746 TI - Isolation and characterization of stable, distinctly bent, trans-chelated bisphosphine palladium(0) species. AB - The novel ligands and (i-Pr-Rucaphos, Cy-Rucaphos) react with (tmeda)PdMe(2) yielding ethane and distinctly bent, trans-spanning bisphosphine chelated 14 electron palladium(0) species, which have been fully characterized, including single crystal X-ray diffraction. PMID- 18414747 TI - NMR transversal relaxivity of aqueous suspensions of particles of Ln(3+)-based zeolite type materials. AB - A series of zeolite-type silicates containing stoichiometric amounts of Ln(3+) ions in the framework (Ln-AV-9 materials), with composition (Na(4)K(2))(Ln(2)Si(16)O(38)).10H(2)O (Ln = Nd, Sm, Eu, Tb, Gd, Dy) has recently been synthesized and characterized. They form paramagnetic microparticles, which as aqueous suspensions have negligible water (1)H longitudinal relaxivities (r(1)) for all Ln(3+) ions studied and quite large transverse relaxivities (r(2)). In this work we further analysed the size distribution of the Ln-AV-9 particles and their r(2)* and r(2) relaxivities. The r(2)* relaxivity effects are explained by the static dephasing regime (SDR) theory. The r(2) relaxivities appear to be strongly dependent on the interval between two consecutive refocusing pulses (tau(CP)) in the train of 180 degrees pulses applied. For long tau(CP) values, the r(2) of the systems saturates at a value, which is always an order of magnitude smaller than r(2)*. These features are explained by a crude model, which takes into account the residual diffusion effect in the static dephasing regime. The large microparticles, although not efficient in T(1) relaxation, are quite effective in enhancing T(2) relaxation, particularly at high magnetic fields. The r(2)* values and the saturation values for r(2) were found to increase linearly with B(0) and mu(0)(2). The largest transversal relaxation rate enhancements were observed for Dy-AV-9 with a saturation value of r(2) of 60 s(-1) mM(-1) and a r(2)* value of 566 s(-1) mM(-1) at 9.4 T and 298 K. PMID- 18414748 TI - Photoluminescent properties and molecular structures of [NaphAu(PPh(3))] and [mu Naph {Au(PPh(3))}(2)] ClO(4) (Naph = 2-naphthyl). AB - The complexes [NaphAu(PPh(3))], and [mu-Naph{Au(PPh(3))}(2)]ClO(4), having the Au C (aromatic) bond have been synthesized and characterized. The unique structure of with two gold atoms bridged by a naphthyl group has been determined by X-ray crystallography. The intramolecular Au-Au separation in is 2.7731(4) A. Upon excitation at 266 nm, both complexes display intraligand phosphorescence at room temperature in solution and in solid state. PMID- 18414749 TI - Supramolecular networks of silver(I) and iron(II) complexes of the third generation tris(pyrazolyl)methane ligand Ph(2)(O)POCH(2)C(pz)(3) (pz = pyrazolyl ring). AB - The new ligand Ph(2)(O)POCH(2)C(pz)(3) (pz = pyrazolyl ring), prepared from the reaction of HOCH(2)C(pz)(3) and Ph(2)P(O)Cl in the presence of base, reacts with either AgBF(4) or Fe(BF(4))(2).6H(2)O in a 2/1 molar ratio to yield {[Ph(2)(O)POCH(2)C(pz)(3)](2)Ag}(BF(4)) () and {[Ph(2)(O)POCH(2)C(pz)(3)](2)Fe}(BF(4))(2) (), respectively. In the structure of , the silver is in an unusual planar geometry with each of the ligands in a kappa(2)-kappa(0) coordination mode. Slow evaporation of a thf solution of yields crystalline [Ph(2)(O)POCH(2)C(pz)(3)Ag](2)(thf)(2)}(BF(4))(2) (). In each cationic unit of , the two Ph(2)(O)POCH(2)C(pz)(3) ligands coordinate to the same two silver(i) centers in a kappa(2)-kappa(1) bonding mode, with a silver atom separation of 3.36 A. The supramolecular structure of both and is dominated by a pair of cooperative hydrogen bonding interactions between the Ph(2)P(O) secondary tecton and a hydrogen atom from a methylene group situated on a neighboring building block, which arranges the cations in chains. The reaction of HC(pz)(3) and AgO(3)SCF(3) (AgOTf) yields {[HC(pz)(3)](2)Ag(2)}(OTf)(2) (). The cationic unit in has a structure very similar to that of , but with a much shorter distance between the silver atoms at 2.86 A. The supramolecular structure of is dominated by an unusual pyrazolyl embrace interaction where the acceptor ring in the C-Hpi interaction is the pyrazolyl ring kappa(1)-bonded to silver in the adjacent dimeric unit rather than the other ring in a kappa(2)-bonded Cpz(2) unit. This interaction arranges the cations in chains which are further organized into sheets by the triflate anions that link the chains via combined AgO/CHO interactions. The iron in is octahedral with each tris(pyrazolyl)methane unit in the kappa(3)-tripodal coordination mode. The supramolecular structure is sheets formed by hydrogen bonding between the Ph(2)P(O) oxygen and a meta-position hydrogen on one of the diphenylphosphine rings from an adjacent cation. PMID- 18414750 TI - Complexes of germanium(IV) fluoride with phosphane ligands: structural and spectroscopic authentication of germanium(IV) phosphane complexes. AB - The first phosphane complexes of germanium(iv) fluoride, trans-[GeF(4)(PR(3))(2)] (R = Me or Ph) and cis-[GeF(4)(diphosphane)] (diphosphane = R(2)P(CH(2))(2)PR(2), R = Me, Et, Ph or Cy; o-C(6)H(4)(PR(2))(2), R = Me or Ph) have been prepared from [GeF(4)(MeCN)(2)] and the ligands in dry CH(2)Cl(2) and characterised by microanalysis, IR, Raman, (1)H, (19)F{(1)H} and (31)P{(1)H} NMR spectroscopy. The crystal structures of [GeF(4)(diphosphane)] (diphosphane = Ph(2)P(CH(2))(2)PPh(2) and o-C(6)H(4)(PMe(2))(2)) have been determined and show the expected cis octahedral geometries. In anhydrous CH(2)Cl(2) solution the complexes are slowly converted into the corresponding phosphane oxide adducts by dry O(2). The apparently contradictory literature on the reaction of GeCl(4) with phosphanes is clarified. The complexes trans-[GeCl(4)(AsR(3))(2)] (R = Me or Et) are obtained from GeCl(4) and AsR(3) either without solvent or in CH(2)Cl(2), and the structures of trans-[GeCl(4)(AsEt(3))(2)] and Et(3)AsCl(2) determined. Unexpectedly, the complexes of GeF(4) with arsane ligands are very unstable and have not been isolated in a pure state. The behaviour of the germanium(iv) halides towards phosphane and arsane ligands are compared with the corresponding silicon(iv) and tin(iv) systems. PMID- 18414751 TI - The coordination chemistry of selenophosphite ligands. Synthesis and characterization of heterometallic tetranuclear clusters [M{CpFe(CO)(2)P(Se)(OR)(2)}(3)](PF(6)) (M = Cu, Ag; R = (n)Pr, (i)Pr) and [Cu(mu X) {CpFe(CO)(2)P(Se)(O(i)Pr)(2)}](2) (X = Cl, Br). AB - A neutral selenium donor ligand, [CpFe(CO)(2)P(Se)(OR)(2)] is used for the construction of Cu(I) and Ag(I) complexes with a well-defined coordination environment. Four clusters [M{CpFe(CO)(2)P(Se)(OR)(2)}(3)](PF(6)), (where M = Cu, R = (n)Pr, ; R = (i)Pr, and M = Ag, R = (n)Pr, ; R = (i)Pr, ) are isolated from the reaction of [M(CH(3)CN)(4)(PF(6))] (where M = Cu or Ag) and [CpFe(CO)(2)P(Se)(OR)(2)] in a molar ratio of 1 : 3 in acetonitrile at 0 degrees C. The reaction of [CpFe(CO)(2)P(Se)(O(i)Pr)(2)] with cuprous halides in acetone produce two mixed-metal, Cu(I)(2)Fe(II)(2) clusters, [Cu(mu-X) {CpFe(CO)(2)P(Se)(O(i)Pr)(2)}](2) (X = Cl, ; Br, ). All six clusters have been fully characterized spectroscopically ((1)H, (13)C, (31)P, and (77)Se NMR, IR), and by elemental analyses. X-Ray crystal structures of and consist of discrete cationic clusters in which three iron-selenophosphito fragments are linked to the central copper or silver atom via selenium atoms. Both clusters and crystallize in the noncentrosymmetric, hexagonal space group P6[combining macron]2c. The coordination geometry around the copper or silver atom is perfect trigonal-planar with Cu-Se and Ag-Se distances, 2.3505(7) and 2.5581(7) A, respectively. X-Ray crystallography also reveals that each copper center in neutral heterometallic clusters and is trigonally coordinated to two halide ions and a selenium atom from the selenophosphito-iron moiety. The structures can also be delineated as a dimeric unit which is generated by an inversion center and has a Cu(2)X(2) parallelogram core. The dihedral angle between the Cu(2)X(2) plane and the plane composed of Cp ring is found to be 24.62 and 84.58 degrees for compound and , respectively. Hence the faces of two opposite Cp rings are oriented almost perpendicular to the Cu(2)X(2) plane in , but are close to be parallel in . This is the first report of the coordination chemistry of the anionic selenophosphito moiety [(RO)(2)PSe](-), the conjugated base of a secondary phosphine selenide, which acts as a bridging ligand with P-coordination on iron and Se-coordination to copper or silver. PMID- 18414752 TI - A ligand-field study of the ground spin-state magnetic anisotropy in a family of hexanuclear Mn(III) single-molecule magnets. AB - A ligand field analysis of two structurally related hexanuclear Mn(iii) coordination complexes reveals that the observed difference in their ground spin state anisotropy originates from the difference in projection coefficients of the single-ion anisotropy to spin states of different total spin quantum-number, S, rather than the geometrical distortions of the metal ions. Furthermore we show that the single-ion second order anisotropy induces fourth and higher order anisotropy terms to the ground spin states of the studied systems, as a consequence of spin-state mixing effects due to the comparable magnitude of the single-ion second order anisotropy and the isotropic exchange parameters. PMID- 18414753 TI - Large-bite bis(phosphite) ligand containing mesocyclic thioether moieties: synthesis, reactivity, group 11 (Cu(I), Au(I)) metal complexes and anticancer activity studies on a human cervical cancer (HeLa) cell line. AB - The large-bite bis(phosphite) ligand [{(-OC(10)H(6)(mu-S)C(10)H(6)O-)P{mu-( OC(10)H(6)(mu-S)C(10)H(6)O-)}P(-OC(10)H(6)(mu-S)C(10)H(6)O-)}] (Pinsertion markP) () was obtained by the reaction of PCl(3) and thiobis(2,2'-naphthol) (). The stoichiometric reactions of with elemental sulfur and selenium afforded the corresponding chalcogenide derivatives [(E)Pinsertion markP(E)] (, E = S; , E = Se) in good yield. Treatment of two equivalents of [ClAu(SMe(2))] with afforded a dinuclear complex [ClAu(Pinsertion markP)AuCl] (), whereas the 1 : 1 reaction with CuI yielded the [(Pinsertion markP)CuI] () complex. The copper(i) complex on treatment with various pyridyl derivatives, produced mixed-ligand complexes [(Pinsertion markP)CuI(NC(5)H(5))] (), [(Pinsertion markP)Cu(2,2'-bpy)]I (), [(Pinsertion markP)Cu(1,10-phen)]I () and {[(Pinsertion markP)Cu(4,4' bpy)]I}(infinity) (). The compounds were tested for their cytotoxic activity on the human cervical cancer (HeLa) cell line. Compounds and were found to inhibit proliferation of HeLa cells significantly. These agents also induced apoptotic cell death in cancer cells. Evidence presented in this study indicated that the compounds and activate the tumor suppressor protein p53 in the colon adenocarcinoma (HCT-116) cell line. PMID- 18414754 TI - Synthesis and structures of boron dihalides supported by the C(6)F(5)-substituted beta-diketiminate ligand [HC(CMe)(2)(NC(6)F(5))(2)](-). AB - The new boron dihalides of the type [HC(CMe)(2)(NC(6)F(5))(2)]BX(2) (X = Cl, Br, I) have been prepared and characterized by single-crystal X-ray diffraction. Of the various synthetic approaches explored, the best method in terms of yield and product purity involves the silylhalide elimination reaction of the silylated iminoamine [HC(CMe)(2)(NC(6)F(5))(N{SiMe(3)}C(6)F(5))] with BX(3). Chloroborenium salt [HC(CMe)(2)(NC(6)F(5))(2)BCl][AlCl(4)] was prepared by treatment of [HC(CMe)(2)(NC(6)F(5))(2)]BCl(2) with AlCl(3) in CH(2)Cl(2) solution. This salt was also structurally authenticated and represents the first such data for a beta diketiminate-supported haloborenium cation. PMID- 18414755 TI - Use of HNO(3) as the source of NO to prepare a nitric oxide complex of ruthenium. AB - Reaction of cis-Ru(bisox)(2)Cl(2), where bisox is 4,4,4',4'-tetramethyl-2,2' bisoxazoline, with HNO(3) in 1 : 4 molar proportion in boiling water under N(2) atmosphere and subsequent addition of an excess of NaClO(4).H(2)O yields [Ru(bisox)(HL)(NO)](ClO(4))(NO(3)) (1). HL is a hydrolysed form of bisox where one of the oxazoline rings opens up. X-Ray crystallography shows that 1 contains an octahedral RuN(5)O core. HL binds the metal through an imino N, an amide N and an alcoholic O atom. Reaction of cis-Ru(bisox)(2)Cl(2) with an excess of NaNO(2) in water gives cis-Ru(bisox)(2)(NO(2))(2) (2). On acidification by HClO(4) in methanol, is smoothly converted to cis-[Ru(bisox)(2)(NO(2))(NO)](ClO(4))(2) (3) due to equilibrium (1). [Formula: see text] (1) The X-ray crystal structures of 2 and 3 have also been determined. NO binds Ru in 1 and 3 linearly. The Ru-NO bond length in 1 is 1.764(13) A and that in 3 is approximately 1.78 A. All the three complexes have been characterised by FTIR, NMR and ESIMS. The NO stretching frequencies in 1 and 3 are 1897 and 1936 cm(-1) respectively. While 3 reverts back to 2 readily in presence of OH(-) [equilibrium (1)], 1 does not react with OH(-). It is concluded that while in the reaction of cis-Ru(bisox)(2)Cl(2) with HNO(3), bisox is hydrolysed following abstraction of NO from HNO(3), generation of the nitrosyl complex 3 via reaction (1) is not accompanied with such hydrolysis. PMID- 18414757 TI - Diimine supported group 10 hydroxo, oxo, amido, and imido complexes. AB - A series of L(2) = diimine (Bian = bis(3,5-diisopropylphenylimino)acenapthene, Bu(t)(2)bpy = 4,4'-di-tert-butyl-2,2'-bipyridine) supported aqua, hydroxo, oxo, amido, imido, and mixed complexes have been prepared. Deprotonation of [L(2)Pt(mu OH)](2)(2+) with 1,8-bis(dimethylamino)naphthalene, NaH, or KOH yields [(L(2)Pt)(2)(mu-OH)(mu-O)](+) as purple (Bian) or red (Bu(t)(2)bpy) solids. Excess KOH gives dark blue [(Bian)Pt(mu-O)](2). MeOTf addition to [(Bu(t)(2)bpy)(2)Pt(2)(mu-OH)(mu-O)](+) gives [(Bu(t)(2)bpy)(2)Pt(2)(mu-OH)(mu OMe)](2+) while [(Bian)Pt(mu-O)](2) yields [(Bian)(2)Pt(2)(mu-OMe)(mu-O)](+). Treatment of [(Bian)Pt(mu-O)](2) with "(Ph(3)P)Au(+)" gives deep purple [(Bian)(2)Pt(2)(mu-O)(mu-OAuPPh(3))](+) while (COD)Pt(OTf)(2) gives a low yield of [(Bian)Pt(3)(mu-OH)(3)(COD)(2)](OTf)(3). Ni(Bu(t)(2)bpy)Cl(2) and [(Ph(3)PAu)(3)(mu-O)](+) in a 3 : 2 ratio yield red [Ni(3)(Bu(t)(2)bpy)(3)(mu O)(2)](2+). M(Bu(t)(2)bpy)Cl(2) (M = Pd, Pt) and [(Ph(3)PAu)(3)(mu-O)](+) give [M(Bu(t)(2)bpy)(mu-OAuPPh(3))](2)(2+) and [Pd(4)(Bu(t)(2)bpy)(4)(mu OAuPPh(3))](3+). Addition of ArNH(2) to [M(Bu(t)(2)bpy)(mu-OH)](2)(2+) (M = Pd, Pt) gives [Pt(2)(Bu(t)(2)bpy)(2)(mu-NHAr)(mu-OH)](2+) (Ar = Ph, 4-tol, 4 C(6)H(4)NO(2)) and [M(Bu(t)(2)bpy)(mu-NHAr)](2)(2+) (Ar = Ph, tol). Deprotonation of [Pt(2)(Bu(t)(2)bpy)(2)(mu-NH-tol)(mu-OH)](2+) with 1,8 bis(dimethylamino)naphthalene or NaH gives [Pt(2)(Bu(t)(2)bpy)(2)(mu-NH-tol)(mu O)](+). Deprotonation of [Pt(Bu(t)(2)bpy)(mu-NH-tol)](2)(2+) with KOBu(t) gives deep green [Pt(Bu(t)(2)bpy)(mu-N-tol)](2). The triflate complexes M(Bu(t)(2)bpy)(OTf)(2) (M = Pd, Pt) are obtained from M(Bu(t)(2)bpy)Cl(2) and AgOTf. Treatment of Pt(Bu(t)(2)bpy)(OTf)(2) with water gives the aqua complex [Pt(Bu(t)(2)bpy)(H(2)O)(2)](OTf)(2). PMID- 18414756 TI - Structure of solvated mercury(II) halides in liquid ammonia, triethyl phosphite and tri-n-butylphosphine solution. AB - Liquid ammonia, trialkyl phosphites, and especially trialkylphosphines, are very powerful electron-pair donor solvents with soft bonding character. The solvent molecules act as strongly coordinating ligands towards mercury(ii), interacting strongly enough to displace halide ligands. In liquid ammonia mercury(ii) chloride solutions separate into two liquid phases; the upper contains tetraamminemercury(ii) complexes, [Hg(NH(3))(4)](2+), and chloride ions in low concentration, while the lower is a dense highly concentrated solution of [Hg(NH(3))(4)](2+) entities, ca. 1.4 mol dm(-3), probably ion-paired by hydrogen bonds to the chloride ions. Mercury(ii) bromide also dissociates to ionic complexes in liquid ammonia and forms a homogeneous solution for which (199)Hg NMR indicates weak bromide association with mercury(ii). When dissolving mercury(ii) iodide in liquid ammonia and triethyl phosphite solvated molecular complexes form in the solutions. The Raman nu(I-Hg-I) symmetric stretching frequency is 132 cm(-1) for the pseudo-tetrahedral [HgI(2)(NH(3))(2)] complex formed in liquid ammonia, corresponding to D(S) = 56 on the donor strength scale. For the Hg(ClO(4))(2)/NH(4)I system in liquid ammonia a (199)Hg NMR study showed [HgI(4)](2-) to be the dominating mercury(ii) complex for mole ratios n(I(-)) : n(Hg(2+)) > or = 6. A large angle X-ray scattering (LAXS) study of mercury(ii) iodide in triethyl phosphite solution showed a [HgI(2)(P(OC(4)H(9))(3))(2)] complex with the Hg-I and Hg-P bond distances 2.750(3) and 2.457(4) A, respectively, in near tetrahedral configuration. Trialkylphosphines generally form very strong bonds to mercury(ii), dissociating all mercury(ii) halides. Mercury(ii) chloride and bromide form solid solvated mercury(ii) halide salts when treated with tri-n-butylphosphine, because of the low permittivity of the solvent. A LAXS study of a melt of mercury(ii) iodide in tri-n-butylphosphine at 330 K resulted in the Hg-I and Hg-P distances 2.851(3) and 2.468(4) A, respectively. The absence of a distinct I-I distance indicates flexible coordination geometry with weak and non-directional mercury(ii) iodide association within the tri-n-butylphosphine solvated complex. PMID- 18414758 TI - Inclusion complexes of the antitumour metallocenes Cp(2)MCl(2) (M = Mo, Ti) with cucurbit[n]urils. AB - The encapsulation of the aquated forms of molybdocene dichloride and titanocene dichloride by cucurbit[n]uril (Q[n], where n = 7 and 8) at different pD values has been studied by (1)H NMR spectroscopy and molecular modelling. (1)H NMR titration experiments indicate that both metallocenes form 1 : 1 host-guest complexes with both Q[7] and Q[8]. In these complexes, both the cyclopentadienyl ligands and metal centre are positioned deep within the cucurbituril cavity. In vitro cell proliferation studies using the cancer cell lines MCF-7 and 2008 showed that the encapsulated molybdocene complex was more active than the corresponding free metallocene, with GI(50) values of 210 and 400 muM respectively. However, unexpectedly the encapsulation of Cp(2)MoCl(2(aq))at pD 7 catalysed significant degradation of the cucurbituril framework in the presence of oxygen. Encapsulation of Cp(2)TiCl(2(aq)) by Q[7] greatly slowed the protonolysis of the cyclopentadienyl ligands in aqueous phosphate buffer (pD 7), while encapsulation in Q[8] only slightly retarded the hydrolytic degradation of the metallocene. PMID- 18414759 TI - Metal-controlled self-assembly of arsenic-vanadium-cluster backbones with organic ligands. AB - Five new organic-inorganic assemblies, [Co(en)(3)][Co(en)(2)As(8)V(14)O(42)(H(2)O)].16H(2)O (1), [Ni(Meen)(3)](4)[Ni(Meen)(2)][As(8)V(14)O(42)(NO(3))](2).8H(2)O (2), [Cd(en)(3)][Cd(phen)(en)(H(2)O)(2)][(en)CdAs(8)V(13)O(41)(H(2)O)].1.5H(2)O (3), [Cd(phen)(2)(en)](2)[(phen)CdAs(8)V(13)O(41)(H(2)O)].21H(2)O.phen (4), [Zn(en)(2)](2)[(bpe)(2)Zn(2)As(8)V(12)O(40)(H(2)O)] (5) (en = ethylenediamine, Meen = 1,2-diaminopropane, phen = 1,10-phenanthroline, and bpe= 1,2-bis(4 pyridyl)ethylene) have been synthesized and characterized. Among them, compounds 1 and 2 are constructed from the [As(8)V(14)O(42)] cluster; compounds 3 and 4 are constructed from the Cd-substituted polyoxovanadate [CdAs(8)V(13)O(41)] cluster, while compound 5 consists of bizinic-substituted polyoxovanadate [Zn(2)As(8)V(12)O(40)] building units. It can be assumed that the metal ions used in these reaction systems play a crucial role in controlling the formation of the arsenic-vanadium-cluster backbones, and further leading to the formation of hybrids based on these clusters. PMID- 18414760 TI - Controllable preparation, network structures and properties of unusual metal organic frameworks constructed from 4,4'-(hexafluoroisopropylidene)diphthalic acid and 4,4'-bipyridyl. AB - Three novel 3D metal-organic frameworks of [Ni(2)(hfpdpt)(bpy)(2)(H(2)O)(2)](H(2)O)(8) (1), [Ni(3)(Hhfpdpt)(2)(bpy)(4)](H(2)O)(2) (2), and [Mn(2)(hfpdpt)(H(2)O)(2)] (3) have been synthesized by the hydrothermal reaction of Ni(II) or Mn(II) nitrate, 4,4' (hexafluoroisopropylidene)diphthalic acid (H(4)hfpdpt), and 4,4'-bipyridyl (bpy). Complex 1 self-assembles into a 3D framework with unconventional 4-connected network topology, while 2 and 3 exhibit two kinds of unusual (4,8)-connected topological networks. Notably, in 1 and 2, the bpy coligand participates in the construction of their 3D frameworks, and the pH value also plays an important role to tune their space arrangements, while in 3, the bpy coligand is not involved in the final crystalline lattice. Furthermore, a detailed investigation on their magnetic and adsorptive properties is carried out. PMID- 18414761 TI - [Multicenter surgical trials recruiting in Germany: current studies]. PMID- 18414762 TI - Detection of toxic metals in waste water from dairy products plant using laser induced breakdown spectroscopy. AB - Laser Induced Breakdown Spectroscopy (LIBS) System was developed locally for determination of toxic metals in liquid samples and the system was tested for analysis of waste water collected from dairy products processing plant. The plasma was generated by focusing a pulsed Nd: YAG laser at 1064 nm on waste water samples. Optimal experimental conditions were evaluated for improving the sensitivity of our LIBS system through parametric dependence investigations. The Laser-Induced Breakdown Spectroscopy (LIBS) results were then compared with the results obtained using standard analytical technique such as Inductively Coupled Plasma Emission Spectroscopy (ICP). The evaluation of the potential and capabilities of LIBS as a rapid tool for liquid sample analysis are discussed in brief. PMID- 18414763 TI - Comparative toxicity of nine metals to two Malaysian aquatic dipterian larvae with reference to temperature variation. AB - A study was conducted to determine the suitability of using selected aquatic dipterian larvae for biomonitoring bioassays. The organisms included a member of the biting midge family that was identified as Culicoides furens and a member of the non-biting midge family, identified as Chironomus plumosus. Median lethal toxicity tests were conducted to observe the variation between metal sensitivities between the two larval forms and how variations in temperature could affect the experimental setup. Nine heavy metals were used in the study. It was observed that the 96 h LC(50) (in mg/L) for the different metals was found to be Zn-16.21 (18.55 +/- 13.87); Cr-0.96 (1.08 +/- 0.84); Ag-4.22 (6.87 +/- 1.57); Ni-0.42 (0.59 +/- 0.25); Hg-0.42 (0.59 +/- 0.25); Pb-16.21 (18.31 +/- 14.11); Cu 42.24 (45.18 +/- 39.30); Mn-4.22 (7.19 +/- 1.25); Cd-0.42 (0.59 +/- 0.25) for the Chironomus plumosus and Zn-4.22 (6.56 +/- 1.88); Cr-0.42 (0.54 +/- 0.30); Ag-0.42 (0.54 +/- 0.30); Ni-0.42 (0.54 +/- 0.30); Hg-0.04 (0.07 +/- 0.01); Pb-0.42 (0.54 +/- 0.30); Cu-42.24 (45.18 +/- 39.30); Mn-4.22 (6.56 +/- 1.88); Cd-0.42 (0.54 +/- 0.30) in the case of the Culicoides furens. With temperature as a variable the LC(50) values were observed to increase from 2.51 mg/L at 10 degrees C to 4.22 ppm at 30 degrees C and to reduce slightly to 3.72 mg/L at 35 degrees C as seen in the case of Zn. It was also observed that at 40 degrees C thermal toxicity and chemical toxicity overlapped as 100% mortality was observed in the controls. This trend was observed in all metals for both C. plumosus and C. furens. Thus indicating temperature played an important role in determining LC(50) values of toxicants. PMID- 18414764 TI - Surgical repair of a vesicovaginal fistula guided by a black braided silk suture line. AB - The purpose of the study was to evaluate the effect of the identification of the margins of a vesicovaginal fistula (VVF) during its repair by passing a black braided silk suture line through the fistula. During the repair of a VVF, a suture line was placed through the fistula. By passing the suture line from the vagina through the fistula tract into the bladder and hence to the outside through the urethral meatus, the circumference of the VVF opening can be easily identified after dissection and excision of the vaginal wall around the fistula opening. There were four VVF patients who underwent a modified Latzko procedure using the thread to guide surgical repair. The operation times and blood loss were 45-90 min and no more than 50 ml, respectively. This trick provides adequate identification of the fistula tract, thus permitting safe and easy fistula repair for a narrow, deep, or atrophic vagina. PMID- 18414765 TI - Leak point pressure does not correlate with incontinence severity or bother in women undergoing surgery for urodynamic stress incontinence. AB - The aim of this study was to correlate the lowest Valsalva or cough leak point pressure (LPP) with clinical measures of incontinence severity and quality of life in women with pure urodynamic stress incontinence (SUI). This is an analysis of the baseline data from a prospective, multicenter, randomized trial comparing the Monarc transobturator sling to the tension-free vaginal tape. One hundred fifty-five women with SUI underwent urodynamic evaluations including abdominal or vesical LPP determinations, and each completed the Sandvik Incontinence Severity Index, a 3-day voiding diary, and quality-of-life questionnaires. In patients with a LPP, there were no significant correlations between LPP and the above clinical measures of incontinence severity or condition-specific quality-of-life questionnaire scores. In this patient population with pure urodynamic SUI, LPP is not a useful urodynamic predictor of baseline SUI severity and its effects on quality of life. PMID- 18414766 TI - Large urethral prolapse formation after calcium hydroxylapatite (Coaptite) injection. AB - A 67-year-old woman who had failed two prior anti-incontinence surgeries presented with stress urinary incontinence and intrinsic sphincteric deficiency. Calcium hydroxylapatite (Coaptite(R)) was injected cystoscopically into the bladder neck and proximal urethra and resolved her incontinence. Seven months later, she presented with difficulty in voiding and a urethral mass. Physical examination revealed a large prolapse of the urethral mucosa obstructing the external urethral meatus. Surgical exploration revealed local migration of calcium hydroxylapatite particles from the site of injection (bladder neck and proximal urethra) to the distal urethra. The prolapsed urethral mucosa was incised and marsupialized. Improper injection techniques likely contributed to urethral prolapse in this complication. Meticulous attention to injection techniques is the key to treatment success of urethral bulking agents, particularly in patients who have a scarred/fixed urethra or have multiple urethral/vaginal surgeries. PMID- 18414767 TI - Three-dimensional visualization of pelvic vascularity. AB - The aim of this study was to establish a 3D digitized model of pelvic vasculature for anatomic study, preoperative planning, and virtual reality. Three adult fresh cadavers were perfused with carboxymethyl cellulose/lead oxide mixture to mark blood vessels, and subjected to multilayer spiral computed tomography scanning to obtain a series of thin sections. Then, the 2D images of the pelvis and pelvic blood vessels were transformed into 3D digitized models using Mimics 11.0. The 2D images of carboxymethyl cellulose/lead oxide filled arteries had the features of entire outline and few constructed defects. The 3D digitized models of the pelvis and pelvic artery system displayed spatial location and the adjacent relationship of arteries with the pelvis. Not only the well-known arteries but also the tiny blood vessels in the reconstructed structures were well demonstrated and observed interactively. The reconstructed tissue flaps, including a lobulated skin flap with the pedicle of superficial epigastric artery, and an iliac flap with the pedicle of deep iliac circumflex artery, demonstrated their blood supply area. This indicated that the modified technique of vascular perfusion with carboxymethyl cellulose/lead oxide and reconstitution with Mimics 11.0 software contributed to 3D digitized model of pelvic vasculature. PMID- 18414768 TI - Supplement use and mortality: the SENECA study. AB - BACKGROUND: It is hypothesis that in relatively healthy older people supplement usage can be consider as healthy life style habit and as such can positively influence longevity. AIM OF THE STUDY: To determine whether supplement use was associated with all-cause mortality in the participants of the SENECA study. METHODS: Baseline measurements were carried out in 1988/1989 among 75 to 80-year old people living in 15 European small towns. All-cause mortality was followed up to April 30, 1999. Data from 920 men and 980 women who were ischemic heart diseases-, stroke- and cancer-free at baseline were included. The multivariate adjusted (for sex, age, years of education, physical activity, BMI, chronic diseases, Mediterranean Diet Score, alcohol use and the place of living) hazard ratio (HRs) and 95% confidence intervals (CIs) of mortality by use of any type of nutrient supplement and by particular nutrient supplement use were estimated by Cox proportional hazards regression models. RESULTS: At baseline, 13% of participants used nutritional supplements, 19% of subjects were smokers. During 10 years of follow-up 445 men and 252 women died. Among non-smokers no significant associations between total supplement use and particular nutrient supplement use were observed. Among smokers use of any type of supplements (Multivariate HR: 1.52; 95%CI: 1.02-2.28), use of vitamin B(1) (Multivariate HR: 1.57; 95%CI: 1.00-2.48) and vitamin B(2) supplements (Multivariate HR: 1.60; 95%CI: 1.00-2.56) were associated with a significantly higher risk of all-cause mortality. The similar tendencies were observed among vitamin B(6) and vitamin C supplement users who were smokers. CONCLUSIONS: Among smokers, participants of the SENECA study, supplement use increased all-cause mortality risk. PMID- 18414769 TI - Dietary lipid intake influences the level of cholesterol bound to haemoglobin in human erythrocytes. AB - BACKGROUND: Blood cholesterol levels are affected by diet and in particular by the type of fat intake. We originally showed that a significant but variable amount of cholesterol is firmly bound to haemoglobin (Hb) yielding the Hb-lipid adduct (Hb-Ch) in erythrocytes isolated from normo-lipidemic males. AIM OF THE STUDY: To establish whether dietary lipids affect the level of Hb-Ch in human erythrocytes. METHODS: Seventy-four healthy free-living adults were separated according to their serum cholesterol levels into two groups: normo cholesterolemic (LDL cholesterol <3.4 mmol/l and total cholesterol <5.2 mmol/l) (NC) and hyper-cholesterolemic (LDL cholesterol >or=3.4 mmol/l) (HC). Habitual dietary information was used to classify subjects in both study groups into sub groups of low-fat (30% total energy as fat). The NC low-fat consumers were placed on a high-lipid (high fat and high-cholesterol) diet whereas the HC subjects with high-fat intake were assigned to a low-lipid (low-fat and low-cholesterol) diet. Both types of dietary intervention were allowed to continue for 6 weeks. The main variable under scrutiny was the Hb-Ch concentration. RESULTS: In both study groups low-fat intake subjects had low levels of Hb-Ch (approx. 0.35 mmol/l RBC) compared with high-fat intake subjects (approx. 0.60 mmol/l RBC), and serum cholesterol was not correlated with Hb-Ch. The two dietary interventions produced substantial changes in the Hb-Ch level that paralleled variation in the serum cholesterol concentration. A high-lipid diet (35% fat, 15% saturated; 580 mg cholesterol) increased Hb-Ch (by approximately 47%, P<0.001) in subjects with low Hb-Ch at onset, whereas a low-lipid diet (28% fat, 9% saturated; 280 mg cholesterol) decreased Hb-Ch (by approximately 40%, P<0.001) in subjects with high Hb-Ch at onset. CONCLUSION: High consumption of dietary lipids, including saturated fat and cholesterol, has an important influence on the level of Hb-Ch in human erythrocytes. PMID- 18414770 TI - Diet revision in overweight children: effect on autonomic and vascular function. AB - In an effort to determine the effect of a 5-month dietary modification on measures of vascular and cardiac autonomic (cANS) function in overweight (OW) children, 15 OW children had standard non-invasive measures of vascular and cANS function assessed pre- and post intervention. Body fat percentage and cANS, but not vascular, function changed significantly after the intervention. Changes in body composition in OW children due to dietary modification alone can result in modest improvements in indices of cardiac risk. PMID- 18414771 TI - Ethnic differences in heart rate: can these be explained by conventional cardiovascular risk factors? AB - OBJECTIVES: South Asians worldwide have an elevated risk of coronary heart disease (CHD), only partly explicable by a higher prevalence of diabetes. Diabetes and dysglycaemia are associated with increased heart rates and abnormal cardiac sympathovagal balance, and are predictive of increased coronary mortality. This study investigated ethnic differences in heart rate, sympathovagal balance, and baroreflex sensitivity (BRS) to establish if they were explained by dysglycaemia/insulin resistance. METHODS: 84 South Asian and 83 European men (age 45-85 yrs), half with established CHD and half without proven CHD, underwent metabolic profiling and assessment of mean RR interval, heart rate variablility (HRV) and BRS. RESULTS: Mean resting RR interval was greater in Europeans than South Asians (1014 (149) ms vs. 936 (158) ms respectively, means (SD); P = 0.005). This corresponded to resting heart rates of 59.1 bpm for Europeans and 64.1 bpm for South Asians. No ethnic differences were detected in HRV or BRS. Insulin resistance, assessed by the homeostasis model of assessment of insulin resistance (HOMA-IR), correlated with mean RR interval (beta coefficient = -0.189, P = 0.04) and BRS (beta coefficient = -0.246, P = 0.006) and similar correlations were seen with fasting serum glucose. However, ethnic differences in mean RR interval persisted after adjustment for age, systolic BP, beta blocker use, and HOMA-IR. INTERPRETATION: South Asian men have higher heart rates compared with European men. This difference was not explained by measures of insulin resistance or other risk factors. This may be indicative of altered sympathovagal balance, and contribute to the greater coronary disease risk in people of South Asian ethnicity. PMID- 18414772 TI - Tuba players reproduce a Valsalva maneuver while playing high notes. AB - Playing wind instruments requires expiratory efforts. Blowing low notes on a tuba means a low resistance to expiration while playing high notes requires a strenuous expiratory strain. The resulting high intrathoracic pressure may reproduce a Valsalva maneuver. Ten tuba players were asked to blow medium loud long (15 seconds) notes at three different pitches (low, middle, and high) and to perform Valsalva maneuvers at 10, 40, and 60 mmHg. Blood pressure (BP) was measured continuously with a Finapres monitor. The four classic phases of the Valsalva maneuver were reproduced with the notes. The expiratory effort produced systolic BP and pulse pressure falls reaching about 24 mmHg with the high note or the Valsalva 60. BP and pulse pressure falls were linearly related to stroke volume reductions. Reflex heart rate (HR) changes were inversely related to BP falls, with maximal increases for the high notes (24 beats/minute) and Valsalva 60 (33 beats/minute). High baroreflex sensitivity was associated with a high HR response. Total peripheral resistance was reflexly elevated to counteract cardiac output reductions. During rebreathing, BP overshoots reached 30 mmHg for the high note and 53 mmHg for the Valsalva 60. Altogether, these findings indicate that blowing notes on a tuba reproduces the cardiovascular changes seen with a Valsalva maneuver with the effects of blowing high notes being close to a classic (40 mmHg) Valsalva maneuver. In addition, the baroreflex sensitivity might be a predictor of the ability to surmount the BP intolerance that could occur during wind instrument playing. PMID- 18414773 TI - Results, side effects and complications after thoracoscopic sympathetic block by clamping. The monza clinical experience. AB - BACKGROUND: Video-endoscopic sympathectomy is the current treatment of choice for severe primary hyperhidrosis. Because of the possible postsurgical side effects, the procedure is carried out using removable endoclips that block sympathetic nerve transmission. This study describes the short and "midterm" side effects and complications of this method for the treatment of palmar, axillary, and facial hyperhidrosis. MATERIALS AND METHODS: Two hundred and ninety-four sympathectomies were carried out between September 2003 and June 2006 and followed-up after 17 months. Patients with isolated facial hyperhidrosis were clamped at the T2-3 level clamping. Patients with isolated axillary hyperhidrosis were clamped at the T3-4 level. RESULTS: All patients with palmar and palmar-axillary hyperhidrosis were completely satisfied after clamping at the third thoracic ganglion (T3) level. Immediate complete recovery was achieved in 98% of patients postsurgery. Major complications were Horner's syndrome (1%) and pneumothorax (3%). Compensatory sweating occurred in 22%. No cases of gustatory sweating were reported. All patients were "satisfied" with their results and no patients requested removal of the clips. INTERPRETATION: Thoracoscopic sympathecotomy clamping is a successful treatment for hyperhidrosis. Local hyperhydrosis does not reoccur after 17 months, but there is some degree of compensatory hyperhidrosis. PMID- 18414774 TI - Transsphenoidal surgery for pituitary adenoma in elderly patients. AB - BACKGROUND: As the population continues to age, the number of elderly patients with symptomatic pituitary tumours will continue to increase. Little information exists as to the safety of pituitary surgery in this patient population. The current study was undertaken to evaluate the presentation and perioperative risk associated with transsphenoidal surgery in patients over the age of 70. METHODS: A retrospective chart review was undertaken for patients over the age of 70 undergoing transsphenoidal surgery at the Penn State Hershey Medical Center between 1995 and 2005. FINDINGS: Sixty-four patients over the age of 70 were identified with an average age of 73.4 years (range 70-84). The average follow-up period was 44 months (range 14-104). Symptoms of mass effect were the presenting complaint in 72% of patients while 9% had documentation of growth on imaging studies. Twelve percent presented with a history consistent with apoplexy. Three patients (5%) presented with acromegaly. Post-operative hospital stay averaged 2.6 days (range 2-7). Eight patients had new hormonal deficits post-operatively (1 ACTH, 3 TSH, 2 ACTH/TSH, 2 vasopressin). There were no cardiopulmonary complications and no deaths within 90 days of surgery. CONCLUSIONS: Transsphenoidal surgery can be performed in patients over the age of 70 without undo significant risks. Surgical removal of pituitary adenomas should be considered the mainstay of treatment in elderly patients in whom treatment is necessary. Patients who are medically suitable candidates should be offered surgical resection as would their younger counterparts. PMID- 18414775 TI - Primary Nocardia farcinica brain abscess with secondary meningitis and ventriculitis in an immunocompetent patient, successfully treated with moxifloxacin. AB - We report a rare case of Nocardia farcinica occipital brain abscess in an immunocompetent patient with no underlying risk factors successfully treated with the antibiotic moxifloxacin. The patient underwent craniotomy and abscess drainage. Initial post-operative treatment with co-trimoxazole produced a limited response. Despite the development of skull base meningitis and ventriculitis subsequent addition of moxifloxacin produced an excellent outcome. PMID- 18414776 TI - Endoscopic trans-nasal approach for biopsy of orbital tumours using image-guided neuro-navigation system. AB - BACKGROUND: Histopathological diagnosis of intraorbital tumours is of crucial value for planning further therapy. The aim of the study was to explore clinical utility of image-guided endoscopy for biopsy of orbital tumours. METHOD: Trans nasal endoscopic biopsy of intraorbital mass lesions was performed in 6 patients using a neuro-navigation system (Medtronic Stealth Station Treon plus). The CT and MRI 1 mm slice images were fused by the system in order to visualise both bony and soft tissue structures. The anatomic fiducial registration protocol was used during the procedure. RESULTS: All lesions were precisely localised and the biopsies could be taken from the representative part of the pathological mass. None of the patients developed aggravation of ocular symptoms after the procedure. The operative corridor as well as the size of orbital wall fenestration could be limited to a minimum. The accuracy of neuro-navigation remained high and stable during the entire procedure. CONCLUSIONS: The image guided neuro-navigation system facilitated endoscopic localisation and biopsy of intraorbital tumours and contributed to the reduction of surgical trauma during the procedure. The technique was particularly useful in small, medially located, retrobulbar tumours and in unclear situations when the structure of the lesion resembled surrounding intraorbital tissue. PMID- 18414777 TI - Impact of state and trait anxiety on the panic response to CCK-4. AB - In order to elucidate the impact of psychological factors on panic severity the correlation between baseline anxiety and panic response to cholecystokinin tetrapeptide (CCK-4), an established model of human anxiety, was investigated in 33 healthy volunteers. Baseline anxiety was assessed with the State-Trait-Anxiety Inventory (STAI). Trait and state anxiety did not differ between panickers and nonpanickers nor were they correlated with panic severity. In conclusion, psychological factors are not major determinants for the subjective panic response to CCK-4 thus emphasising the importance of neurobiological factors. PMID- 18414778 TI - Genome-wide linkage analysis for circulating levels of adipokines and C-reactive protein in the Quebec family study (QFS). AB - Adipose tissue synthesizes and secretes a wide range of biologically active molecules considered as inflammatory markers whose dysregulation in obesity plays a role in the development of insulin resistance and vascular disorders. Thus, finding genes that influence circulating levels of inflammatory biomarkers may provide insights into the genetic determinants of obesity-related metabolic diseases. We performed linkage analyses for fasting plasma levels of adiponectin, C-reactive protein (CRP), interleukin-6 (IL-6) and tumor-necrosis factor-alpha (TNF-alpha) in 764 subjects enrolled in the Quebec family study (QFS). A maximum of 393 pairs of siblings from 211 nuclear families were available for analyses. A total of 443 markers spanning the 22 autosomal chromosomes with an average inter marker distance of 6.24 Mb were genotyped. Linkage was tested using both allele sharing (SIBPAL) and variance component linkage methods (MERLIN). We showed suggestive evidence of linkage for plasma adiponectin levels on chromosome 15q21.1 [D15S659; logarithm of the odds (LOD) score = 2.23], 3q13.33 (D3S3023; LOD = 2.09), 20q13.2 (D20S197; LOD = 1.96) and 14q32.2 (D14S1426; LOD = 1.79). Evidence of linkage (SIBPAL) was also found for CRP on 12p11.23 (P = 0.001) and 12q15 (P = 0.0005) and for IL-6 on 14q12 (P = 0.002). None of these linkages remained significant after adjustment for body mass index. No evidence of linkage was found for TNF-alpha plasma levels. These results suggest that several QTLs can influence plasma levels of adiponectin and CRP, partly via their effects on adiposity. PMID- 18414779 TI - Subsequent pregnancy outcomes in recurrent miscarriage patients with a paternal or maternal carrier of a structural chromosome rearrangement. AB - Information concerning the prognosis of subsequent pregnancies in patients with reciprocal translocations is limited. This study was performed to determine the percentage success rate with first pregnancies after ascertainment of a carrier status. A total of 2,382 couples with a history of two or more consecutive miscarriages were studied in multicenters. The prevalence of an abnormal chromosome in either partner was examined, and subsequent success rates were compared between cases with and without an abnormal karyotype in either partner. A total of 129 couples (5.4%) had an abnormal karyotype in one partner excluding inversion 9 in 44 men and in 85 women. Thus, 2,253 couples had a normal karyotype in both partner. Eighty-five (3.6%) had translocations, 13 being Robertsonian translocations. Twenty-nine of the 46 cases (63.0%) who became pregnant with reciprocal translocations in either partner experienced a live birth with natural conception. In contrast, 950 of 1,207 cases (78.7%) with normal chromosomes had successful live births, the difference being significant (P = 0.019). No infant with an unbalanced translocation was found in 29 cases of successful pregnancy following recurrent miscarriage. Pregnancy prognosis was worsened with either maternal or paternal reciprocal translocations. Explanation of the success rate with natural conception should be provided before the subsequent pregnancy after ascertainment of carrier status. PMID- 18414780 TI - Genotype and phenotype analyses in 136 patients with single large-scale mitochondrial DNA deletions. AB - We examined 136 patients with mitochondrial DNA (mtDNA) deletion. Clinical diagnoses included chronic progressive external ophthalmoplegia (94 patients); Kearns-Sayre syndrome (KSS; 33 patients); Pearson's marrow-pancreas syndrome (six patients); and Leigh syndrome, Reye-like syndrome, and mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like episodes (one patient). The length and location of deletion were highly variable. Only one patient had deletion within the so-called shorter arc between the two origins of mtDNA replication. The length of deletion and the number of deleted transfer ribonucleic acid (tRNAs) showed a significant relationship with age at onset. Furthermore, KSS patients had longer and larger numbers of deleted tRNAs, which could be risk factors for the systemic involvement of single mtDNA deletion diseases. We found 81 patterns of deletion. Direct repeats of 4 bp or longer flanking the breakpoints were found in 96 patients (70.5%) and those of 10 bp or longer in 49 patients (36.0%). We found two other common deletions besides the most common deletion (34 patients: 25.0%): the 2,310-bp deletion from nt 12113 to nt 14421 (11 patients: 8.0%) and the 7,664-bp deletion from nt 6330 to nt 13993 (ten patients: 7.3%). These deletions had incomplete direct repeats longer than 13 bp with one base mismatch. PMID- 18414781 TI - Analysis of regulatory polymorphisms in organic ion transporter genes (SLC22A) in the kidney. AB - Organic cation transporters (OCTs) and organic anion transporters (OATs) (SLC22A family) play crucial roles in the renal secretion of various drugs. Messengar ribonucleic acid (mRNA) expression of transporters can be a key factor regulating interindividual differences in drug pharmacokinetics. However, the source of variations in mRNA levels of transporters is unclear. In this study, we focused on single nucleotide polymorphisms (SNP) in the promoter region [regulatory SNPs (rSNPs)] as candidates for the factor regulating mRNA levels of SLC22A. We sequenced the promoter regions of OCT2 and OAT1-4 in 63 patients and investigated the effects of the identified rSNPs on transcriptional activities and mRNA expression. In the OCT2 promoter region, one deletion polymorphism (-578_ 576delAAG) was identified; -578_-576delAAG significantly reduced OCT2 promoter activity (p < 0.05), and carriers of -578_-576delAAG tend to have lower OCT2 mRNA levels, but the difference is not significant. There was no rSNP in the OAT1 and OAT2 genes. The five rSNPs of OAT3 and one rSNP of OAT4 were unlikely to influence mRNA expression and promoter activity. This is the first study to investigate the influences of rSNPs on mRNA expression of SLC22A in the kidney and to identify a regulatory polymorphism affecting OCT2 promoter activity. PMID- 18414782 TI - Mutational analysis of the breast cancer susceptibility gene BRIP1 /BACH1/FANCJ in high-risk non-BRCA1/BRCA2 breast cancer families. AB - The BRIP1 gene encodes a helicase interacting with BRCA1, which contributes to BRCA1-associated DNA repair function. Germ-line BRIP1 mutations affecting the helicase domain activity have been identified in early onset breast cancer patients. In addition, BRIP1 was recently identified as deficient in Fanconi anemia (FA) complementation group J. Given the growing evidence now linking BRCA1, BRCA2, and the FA pathway, as well as the involvement of FA proteins (BRCA2/FANCD1 and PALB2/FANCN) in breast cancer susceptibility, we sought to evaluate the contribution of FANCJ gene alterations regarding breast cancer susceptibility among our cohort of 96 breast cancer individuals from high-risk non-BRCA1/2 French Canadian families. No deleterious mutation, exon deletion, or retention of intronic portions could be identified. However, extensive analysis of the promoter and whole exonic and flanking intronic regions of FANCJ led to the identification of 42 variants, including 22 novel variants not previously reported, four of which were located in the promoter region. Transcription factors analysis revealed a potential involvement of FANCJ promoter variants in regulation of FANCJ expression, and reporter gene assays were performed. The allelic frequency was assessed in a cohort of 73 unaffected French Canadian individuals, and haplotype analysis and tagging single nucleotide polymorphism (SNP) identification were also performed. Although our study unlikely involves FANCJ as a high-risk predisposition gene in non-BRCA1/2 high-risk French Canadian families, the possible association of FANCJ missense variants with phenotypes associated with FA, such as childhood cancer, cannot be excluded. PMID- 18414783 TI - A case of rheumatoid arthritis complicated by demyelination in both cerebral cortex and spinal cord during etanercept therapy. AB - Tumor necrosis factor (TNF) antagonists, including etanercept, have been approved for the treatment of rheumatoid arthritis (RA). These agents are not free of adverse events like other antirheumatic agents. Several important adverse events in CNS lesions have been reported. In this paper, we report on one patient with RA that had complications from a demyelinating disorder during TNF-blockade therapy using etanercept at 24 months after initial administration. A 66-year-old Japanese woman was diagnosed with RA in 1959. She received various disease modifying antirheumatic drugs (DMARDs), but all of these agents were ineffective. She was administered etanercept in June 2005, and stayed well. Twenty-four months after the initial administration of etanercept, she developed palsy of bilateral upper extremities and gait disturbance subacutely, and was then admitted to our institute in August 2007. MRI of her spinal cord revealed a high-intensity lesion from the third through to the seventh cervical (C3-C7) levels. Additionally, T2 weighted MRI images showed disseminated high-intensity lesions in the white matter of brain. She was suspected of having a demyelinating disorder based on these MRI findings. There was no significant finding that pointed to another neurological disorder. High-dose corticosteroid therapy was conducted and was effective for her. PMID- 18414784 TI - Treatment of upper cervical spine involvement in rheumatoid arthritis patients. AB - The cervical spine, especially the upper cervical spine, is a common focus of destruction by rheumatoid arthritis (RA). Because of its potentially debilitating and life-threatening sequelae, cervical spine involvement remains a priority in the diagnosis and treatment of RA. Many studies show that early surgical intervention gives a more satisfactory outcome. Surgery aims to establish spinal stability and to prevent neurological deterioration and injury to the spinal cord, leading to improved neurological function. The recent sophisticated screw rod-plate technique allows one to obtain a solid fixation of the upper cervical spine with a high possibility of bone union even in RA patients. Although surgery of the occipitoatlantoaxial region is a challenge with many possibilities of serious complications, recent advances in the surgical technique, complete understanding of the anatomy, and precise preoperative evaluation have decreased complication rates. Early consultation with a specialized spine surgeon is mandatory once cervical involvement is suspected in an RA patient because once the patient becomes myelopathic, the rate of long-term mortality increases and the chance of neurological recovery decreases. PMID- 18414785 TI - Cross-cultural adaptation and validation of the Japanese version of the Childhood Health Assessment Questionnaire (CHAQ). AB - To assess cross-cultural adaptation, and to validate the parent's version of a health-related quality-of-life instrument, the Childhood Health Assessment Questionnaire (CHAQ) was investigated after its translation into Japanese. A total of 132 subjects were enrolled: 63 patients with juvenile idiopathic arthritis (JIA) (34 systemic and 29 polyarticular) and 69 healthy children. The CHAQ distinguished clinically between healthy subjects and the two JIA subtypes of patients. The average disability index (DI) scores for systemic JIA (sJIA) and polyarticular JIA (pJIA) patients and healthy subjects were 1.5, 1.2, and 0.0, respectively. All variables in the questionnaire were shown to be significant (P < 0.001). Patients with pJIA showed better correlation than those with sJIA. Significant correlation was seen in the polyarticular group with CRP, ESR, parents' VAS, the number of joints with pain, and the number of active joints. However, there was even a negative correlation between DI and parent's assessment of overall well-being for the sJIA group. The Japanese version of the CHAQ was a reliable and valid tool for the functional assessment of children with pJIA. Functional ability, as assessed by the CHAQ, may not be the main consideration of sJIA patients' parents when assessing their child's status. PMID- 18414786 TI - On the deflection of light by topological defects in nematic liquid crystals. AB - The influence of controllable parameters like temperature and wavelength on the trajectories of light in a nematic liquid crystal with topological defects is studied through a geometric model. The model incorporates phenomenological details as how the refractive indices depend on such parameters. The deflection of light by the topological defect is then shown to be greater at lower temperatures and shorter wavelengths. PMID- 18414787 TI - Wetting transition of a nematic liquid crystal on a periodic wedge-structured substrate. AB - It is known that the wetting behaviour of a fluid is deeply altered by the presence of rough or structured substrates. We first review some simple considerations about isotropic fluids and rough substrates, and then we generalize Wenzel's law, which assigns an effective contact angle to a droplet on a rough substrate, when the wetting layer has an ordered phase, like a nematic. We estimate the conditions for which the wetting behavior of an ordered fluid can be qualitatively different from that usually found in a simple fluid. To support our general considerations, we use the Landau-de Gennes mean field approach to investigate theoretically and numerically the wetting transition of a nematic phase on a periodic triangular structured substrate. PMID- 18414788 TI - Layered TiO2: PVK nano-composite thin films for photovoltaic applications. TiO2: PVK nano-composite thin films. AB - The influence of the solvent used for spin-coating on the homogeneity of poly(N vinylcarbazole) (PVK) films is investigated. Homogenous films are obtained only by the use of toluene, solution in tetrahydrofuran (THF) and chloroform results in radially oriented inhomogeneities and films prepared by use of N methylpyrrolidone and dimethylacetamide show particle formation during spin coating. Layered nano-composite thin films are prepared by spin-coating a PVK film on top of a nano-structured titanium dioxide ( TiO2) layer. The TiO2 thin films are prepared by a sol-gel process using an amphiphilic copolymer as structure-directing agent. Structural characterisation of the TiO2 :PVK nano composite films is done by field emission scanning electron microscopy (FESEM) and grazing-incidence small-angle scattering (GISAXS). Bare TiO2 films are probed for comparison. Light is basically only absorbed in the ultraviolet regime and absorption slightly increases upon addition of PVK, which makes the layered TiO2 :PVK nano-composite thin films good candidates for UV photovoltaic devices. Furthermore, absorption remains stable over a period of several days. PMID- 18414790 TI - Cerebellar anaplastic astrocytoma in a teenager with Ollier Disease. AB - INTRODUCTION: Ollier Disease is a sporadic skeletal disorder with a predisposition to oncogenesis. It is estimated at around 1/100,000. We are presenting a young patient with Ollier Disease and high-grade astrocytoma. CASE REPORT: A 14-year-old, Caucasian male with Ollier Disease presented with a history of headaches, vomiting, blurred vision, and unsteady gait. Brain MRI with contrast showed a 41 x 55 mm mass in the posterior fossa with spotty enhancement, which pathology proved to be anaplastic astrocytoma. CONCLUSION: Despite the universal acceptance that Ollier Disease carries a high risk of developing malignancy there is very little in the literature about systematic screening. We recommended a cost-effective screening regime for these patients. PMID- 18414789 TI - Cytosine arabinoside affects the heat and capsaicin receptor TRPV1 localisation and sensitivity in human sensory neurons. AB - BACKGROUND: Cytosine arabinoside (Ara C) is a useful chemotherapy agent, used for treating acute myeloid leukaemia, although it may be associated with side effects including painful neuropathy. It is also used for in vitro neuronal studies to limit the proliferation of non-neuronal cells and thereby select nondividing neuronal cells. We studied the effects of Ara C on human dorsal root ganglion (DRG) neurons, especially the expression and sensitivity of the ion channel TRPV1, which responds to noxious heat and capsaicin and is a key mediator of neuropathic pain. METHODS: Human DRG neurons were cultured with or without Ara C for 2 weeks, after which Ara C was discontinued. Double immunostaining for the regenerative neuronal marker Gap43 and the capsaicin receptor TRPV1 showed that the normal membrane-bound localisation of TRPV1 was absent in neurons with Ara C treatment, and as expected there was massive diminution of dividing non-neuronal cells. Calcium imaging studies showed that during exposure to Ara C the neurons lost responsiveness to capsaicin, although ionomycin responses were intact, indicating general cell viability and responsiveness. Between 2 days and up to 3 weeks after removal of Ara C, the neuronal responses to capsaicin were regained and were observed to be four times (P = 0.0008, Student's t-test) that of controls, but there was only a gradual recovery of non-neuronal cells. Three to six weeks after Ara C removal, capsaicin responses were comparable to controls. CONCLUSIONS: It is postulated that Ara C treatment blocked insertion of TRPV1 in the cell membrane, resulting in accumulation of the receptors in the cytoplasm, loss of capsaicin sensitivity, and membrane-bound immunostaining, which was restored with a rebound on withdrawal of Ara C. The observed pattern of loss of capsaicin sensitivity, followed by hypersensitivity and recovery, appears to reflect some of the features observed in chemotherapy-induced neuropathy, and may provide a model for developing new treatments and prophylaxis. PMID- 18414791 TI - Co-fermentation of grape must by Issatchenkia orientalis and Saccharomyces cerevisiae reduces the malic acid content in wine. AB - Grape must was fermented by a mixed culture of Saccharomyces cerevisiae W-3 (a wine yeast) and Issatchenkia orientalis KMBL 5774 (a malic acid-degrading yeast). Co-fermentation with 1:1 (v/v) inoculum ratio of W-3 and KMBL 5774 decreased malic acid to 0.33 mg/ml from 1.1 mg ml with W-3 alone. Ethanol production was the same in both cases (7.8%, v/v). Acetaldehyde, 1-propanol, 2-butanol and isoamyl alcohol all decreased, with an increase in methanol, in the co-fermented wine. Sensory evaluation showed a higher score in the wine fermented with 1:1 (v/v) inoculum ratio than those obtained by 4:1 (v/v) inoculum ratio or W-3 alone. PMID- 18414792 TI - Novel 3-dimensional bioelectrode for mediatorless bioelectrochemical denitrification. AB - A novel bioelectrochemical method for denitrification was developed using electricity as the electron donor. The novel electrode contained both Ochrobactrum anthropi SY509, which was permeabilized as a biocatalyst, and copper powder as a conducting material. Using this electrode, a high denitrification efficiency of 1 mmol N-NO(3)(-)/g dry cell x h was achieved via direct electron transfer without using mediator. PMID- 18414793 TI - High-cell-density cultivation of Schizochytrium sp. in an ammonium/pH-auxostat fed-batch system. AB - Thraustochytrids, in particular Schizochytrium spp., are used for the production of the valuable polyunsaturated fatty acid, docosahexaenoic acid (DHA; 22:6 n-3). Growth of Schizochytrium sp. G13/2S in a defined medium was initially made in shake-flask cultures to determine the optimum concentrations of glucose (100-200 g l(-1)) and ammonia ( approximately 300 mg l(-1)) that could be used by this microorganism. In subsequent fermenter cultures, a pH-auxostat method was used to maintain NH(3) from 200-300 mg l(-1). During the first 49 h of fermentation, 150 g glucose l(-1) produced 63 g cell dry wt l(-1). Although growth was not limited by the supply of nitrogen, total fatty acids were at 25% cell dry wt which is more than half the final lipid content of commercially-grown Schizochytrium biomass which uses N-limited medium in the final stages for maximum lipid accumulation. This strategy is therefore useful for the cultivation of Schizochytrium to a high cell density up to the point when lipid accumulation can be triggered by N exhaustion. PMID- 18414794 TI - Purification of alpha-galactosidase and invertase by three-phase partitioning from crude extract of Aspergillus oryzae. AB - Alpha-galactosidase and invertase were accumulated in a coherent middle phase in a three-phase partitioning system under different conditions (ammonium sulphate, ratio of tert-butanol to crude extract, temperature and pH). Alpha-galactosidase and invertase were purified 15- and 12-fold with 50 and 54% activity recovery, respectively. The fractions of interfacial precipitate arising from the three phase partitioning were analyzed by SDS-PAGE. Both purified preparations showed electrophoretic homogeneity on SDS-PAGE. PMID- 18414795 TI - Comparison between Escherichia coli K-12 strains W3110 and MG1655 and wild-type E. coli B as platforms for xylitol production. AB - Escherichia coli W3110 was previously engineered to produce xylitol from a mixture of glucose plus xylose by expressing xylose reductase (CbXR) and deleting xylulokinase (DeltaxylB), combined with either plasmid-based expression of a xylose transporter (XylE or XylFGH) (Khankal et al., J Biotechnol, 2008) or replacing the native crp gene with a mutant (crp*) that alleviates glucose repression of xylose transport (Cirino et al., Biotechnol Bioeng 95:1167-1176, 2006). In this study, E. coli K-12 strains W3110 and MG1655 and wild-type E. coli B were compared as platforms for xylitol production from glucose-xylose mixtures using these same strategies. The engineered strains were compared in fed-batch fermentations and as non-growing resting cells. Expression of CRP* in the E. coli B strains tested was unable to enhance xylose uptake in the presence of glucose. Xylitol production was similar for the (crp*, DeltaxylB)-derivatives of W3110 and MG1655 expressing CbXR (average specific productivities of 0.43 g xylitol g cdw( 1 ) h(-1) in fed-batch fermentation). In contrast, results varied substantially between different DeltaxylB-derivative strains co-expressing either XylE or XylFGH. The differences in genetic background between these host strains can therefore profoundly influence metabolic engineering strategies. PMID- 18414796 TI - Immobilization of proteins to biacore sensor chips using Staphylococcus aureus sortase A. AB - The immobilization of proteins to surfaces is an active area of research due to strong interest in protein-based sensors. Here, we describe a novel method for immobilizing ligand proteins onto Biacore sensor chips using the transpeptidase activity of Staphylococcus aureus sortase A (SrtA). This method provides a robust and gentle approach for the site-directed, covalent coupling of proteins to biosensor chips. Notably, the high specificity of the sortase allows immobilization of proteins from less than pure protein samples allowing short cuts in protein purification protocols. PMID- 18414797 TI - Functional characterization of a putative endoglucanase gene in the genome of Zymomonas mobilis. AB - The sequence of the putative endoglucanase gene ZMO1086 in the genome of Zymomonas mobilis showed a 40% similarity with known bacterial endoglucanase genes. The upstream region of this putative gene revealed the presence of characteristic promoter (-10 and -35 regions) and a Shine-Dalgarno region. The putative endoglucanase gene was poorly expressed from the native promoter of Z. mobilis and therefore the putative endoglucanase gene was cloned and expressed in Escherichia coli BL21. The overexpressed gene product CelA was purified to homogeneity and the optimal activity was observed at 30 degrees C and pH 6 respectively. PMID- 18414798 TI - Degradation of cellulose by the major endoglucanase produced from the brown-rot fungus Fomitopsis pinicola. AB - An endoglucanase that is able to degrade both crystalline and amorphous cellulose was purified from the culture filtrates of the brown-rot fungus Fomitopsis pinicola grown on cellulose. An apparent molecular weight of the purified enzyme was approximately 32 kDa by SDS-PAGE analysis. The enzyme was purified 11-fold with a specific activity of 944 U/mg protein against CMC. The partial amino acid sequences of the purified endoglucanase had high homology with endo-beta-1,4 glucanase of glycosyl hydrolase family 5 from other fungi. The K(m) and K(cat)values for CMC were 12 mg CMC/ml and 670/s, respectively. The purified EG hydrolyzed both cellotetraose (G4) and cellopentaose (G5), but did not degrade either cellobiose (G2) or cellotriose (G3). PMID- 18414799 TI - An improved method for Beauveria bassiana transformation using phosphinothricin acetyltransferase and green fluorescent protein fusion gene as a selectable and visible marker. AB - An improved transformation method for the biocontrol agent, Beauveria bassiana, was developed. For convenience of transformation selection and detection, the coding regions of the genes for phosphinothricin acetyltransferase and green fluorescent protein were fused and an expression vector, pBFT, carrying this fusion was constructed. Under optimum conditions, over 60 transformants microg( 1) plasmid DNA were obtained. B. bassiana conidia frozen 1 month at -80 degrees C were fully competent for transformation. The method was significantly less laborious and more rapid than current methods for B. bassiana. The bar::egfp provides a selectable and visible marker which may expedite future genetic engineering of this fungus. PMID- 18414800 TI - Engineering of ligand specificity of periplasmic binding protein for glucose sensing. AB - A novel glucose-sensing molecule was created based on galactose/glucose-binding protein (GGBP). GGBP mutants at Asp14, a residue interacting with the 4th hydroxyl group of the sugar molecule, were constructed by mutagenesis to improve the ligand specificity of GGBP. The autofluorescence-based analysis of the binding abilities of these engineered GGBPs showed that the GGBP mutants Asp14Asn and Asp14Glu bound only to glucose in a concentration-dependent manner, without being affected by the presence of galactose. The Phe16Ala mutation, which leads to an increase in the K (d) value toward glucose, was then introduced into these two glucose-specific mutant GGBPs. One of the constructed GGBP double-mutants, Asp14Glu/Phe16Ala, had a glucose specificity with a K(d) value of 3.9 mM, which makes it suitable for use in the measurement of the physiological glucose concentration. Our results demonstrate that it is possible to construct a GGBP which specifically recognizes glucose and has a higher K(d) value and use it as a molecular recognition element of blood glucose monitoring systems by combining two different mutations based on the 3D structure of GGBP. PMID- 18414801 TI - Cloning, sequencing and expression of a dextransucrase gene (dexYG) from Leuconostoc mesenteroides. AB - The gene dexYG encoding the dextransucrase from an industrial strain of Leuconostoc mesenteroides 0326 was isolated by PCR. The nucleotide sequence of the dexYG gene consists of an open reading frame (ORF) of 4,584 bp, coding for a 1,527 aa protein with a Mr of 170 kDa. The results were analysed by a BLAST similarity search of the GenBank database, which revealed the amino acid sequence was similiar to dsrD derived from L. mesenteroides Lcc4. The dexYG gene was subcloned into the plasmid pET28a(+) and was expressed in E. coli BL21 (DE3) by IPTG induction. The pH value was one of the main reasons which caused the degradation of enzyme activity in the later stage of induction. The highest activity was reached 36 U/ml after 5 h induction in medium at pH 6.0. Biotransformation yield of the enzyme reached 65% and the molecular weight of transformed dextran was more than 68 kDa in 2 h. PMID- 18414802 TI - Identification of alkane hydroxylase genes in Rhodococcus sp. strain TMP2 that degrades a branched alkane. AB - Rhodococcus sp. TMP2 is an alkane-degrading strain that can grow with a branched alkane as a sole carbon source. TMP2 degrades considerable amounts of pristane at 20 degrees C but not at 30 degrees C. In order to gain insights into microbial alkane degradation, we characterized one of the key enzymes for alkane degradation. TMP2 contains at least five genes for membrane-bound, non-heme iron, alkane hydroxylase, known as AlkB (alkB1-5). Phylogenetical analysis using bacterial alkB genes indicates that TMP2 is a close relative of the alkane degrading bacteria, such as Rhodococcus erythropolis NRRL B-16531 and Q15. RT-PCR analysis showed that expressions of the genes for AlkB1 and AlkB2 were apparently induced by the addition of pristane at a low temperature. The results suggest that TMP2 recruits certain alkane hydroxylase systems to utilize a branched alkane under low temperature conditions. PMID- 18414803 TI - Poly(vinyl alcohol) acetoacetate-based tissue adhesives are non-cytotoxic and non inflammatory. AB - Polymer-based tissue adhesives composed of poly(vinyl alcohol) acetoacetate (PVOH acac) and cross-linking amines were investigated for their effects on cell survival and inflammatory cell activation using in vitro mouse cell cultures. Cytotoxicity of tissue adhesives was evaluated by placing adhesives in direct contact with 3T3 fibroblast cells. Tissue adhesives formulated from PVOH acac and 3-aminopropyltrialkoxysilane (APS) were non-cytotoxic to fibroblasts; adhesives formulated from PVOH acac and aminated poly(vinyl alcohol) (PVOH amine) were also non-cytotoxic to fibroblasts. In contrast, a commercial adhesive composed of 2 octyl cyanoacrylate was highly cytotoxic to fibroblasts. The inflammatory potential of tissue adhesives was evaluated by exposing J774 macrophage cells to adhesives, and measuring TNF-alpha release from macrophages. PVOH acac-based tissue adhesives did not elicit inflammatory TNF-alpha release from macrophages. These results suggest that PVOH acac-based tissue adhesives are non-cytotoxic and non-inflammatory. Such tissue adhesives represent a promising technology for a variety of medical applications, including surgical wound closure and tissue engineering, and the results are also significant in the design of in vitro cell culture systems to study biomaterials. PMID- 18414804 TI - One-step purification and characterization of an intracellular beta-glucosidase from Metschnikowia pulcherrima. AB - A collection of 60 non-Saccharomyces yeasts isolated from grape musts in Uruguayan vineyards was screened for beta-glucosidase activity and Metschnikowia pulcherrima was the best source of this enzyme activity. Its major beta glucosidase was successfully purified to homogeneity by ion-exchange chromatography on amino-agarose gel. The enzyme exhibited an optimum catalytic activity at 50 degrees C and pH 4.5 and was active against (1 --> 4)-beta and (1 -> 2)-beta glycosidic linkages. In spite of preserving 100% of its activity and stability in the presence of 12% (v/v) ethanol and 5 g glucose/l, the enzyme was unstable below pH 4. We characterized the beta-glucosidase from M. pulcherrima with a view to its potential applications in wine-making. PMID- 18414805 TI - Microbial fuel cell technology for measurement of microbial respiration of lactate as an example of bioremediation amendment. AB - Microbial fuel cell (MFC) based sensing was explored to provide for the development of an in situ bioremediation monitoring approach for substrate concentrations and microbial respiration rates. MFC systems were examined in column systems where Shewanella oneidensis MR1 used an external electron acceptor (an electrode) to metabolize lactate (a bioremediation additive) to acetate. Column systems were operated with varying influent lactate concentrations (0-41 mM) and monitored for current generation (0.01-0.39 mA). Biological current generation paralleled bulk phase lactate concentration both in the influent and in the bulk phase at the anode; current values were correlated to lactate concentration at the anode (R(2) = 0.9), The electrical signal provided real-time information for electron donor availability and biological activity. These results have practical implications for efficient and inexpensive real-time monitoring of in situ bioremediation processes where information on substrate concentrations is often difficult to obtain and where information on the rate and nature of metabolic processes is needed. PMID- 18414807 TI - In situ recovery of the aroma compound perillene from stirred-tank cultured Pleurotus ostreatus using gas stripping and adsorption on polystyrene. AB - Supplementation of the key metabolite, alpha-(Z)-acaridiol, to stirred-tank cultured Pleurotus ostreatus was used to demonstrate that integrated in situ product recovery resulted in high conversion rates and quantitative separation of the target product perillene from the nutrient medium. The conversion of beta myrcene by P. ostreatus was scaled-up from shake-flasks into a controlled, stirred tank bioreactor equipped with gas stripping and adsorption on a polystyrene fixed bed. The formation of the attractive flavour compound perillene was measured daily using standard controlled capillary gas chromatography. The formation of alpha-(Z)-acaridiol was the metabolic bottleneck of the conversion of beta-myrcene to perillene. Efficient in situ recovery of the volatile product enabled quantitative separation of the pure flavour compound. Appropriated bioprocessing, i.e. in situ separation of product, steadily shifted the metabolic equilibria and thus accomplished high conversion rate and pure product. PMID- 18414806 TI - Overexpression of phytoene synthase gene from Salicornia europaea alters response to reactive oxygen species under salt stress in transgenic Arabidopsis. AB - A phytoene synthase gene SePSY was isolated from euhalophyte Salicornia europaea L. The 1655 bp full-length SePSY has an open reading frame of 1257 bp and encodes a 419-amino acid protein. The overexpression of SePSY enhanced the growth of transgenic Arabidopsis. When the plants were exposed to 100 mM NaCl, the photosynthesis rate and photosystem II activity (Fv/Fm) increased from 92% to 132% and from 9.3% to 16.6% in the transgenic lines than in the wild-type, respectively. The transgenics displayed higher activities of SOD and POD and lower contents of H(2)O(2) and MDA than the WT. In conclusion, the transgenic lines showed higher tolerance to salt stress than WT plants by increased photosynthesis efficiency and antioxidative capacity. This is the first report about improving the salt tolerance by genetic manipulation of carotenoid biosynthesis. PMID- 18414808 TI - Role of whole-body [18F] fluorodeoxyglucose positron emission tomography/computed tomography (FDG-PET/CT) and conventional techniques in the staging of patients with Hodgkin and aggressive non Hodgkin lymphoma. AB - PURPOSE: The aim of this study was to evaluate the role of [(18)F]fluorodeoxyglucose positron emission tomography/computed tomography (FDG PET/CT) in the staging of Hodgkin's and aggressive non-Hodgkin's lymphoma (HL and NHL), comparing it with conventional diagnostic methods, i.e. contrast-enhanced CT and bone marrow biopsy. MATERIALS AND METHODS: Sixty-five consecutive patients (30 HL and 35 NHL) who underwent conventional disease staging and FDG-PET/CT were included. Concordance between conventional methods and PET was established when both procedures identified the same disease stage. Discordant findings were investigated further by using other diagnostic techniques (ultrasonography or magnetic resonance imaging) and/or clinical follow-up. RESULTS: PET correctly staged 93.8% of enrolled patients (61/65), whereas conventional techniques correctly staged 89.2% (58/65; p=NS, Chi(2) test). There was complete concordance in 54/65 patients (83.1%); among the remaining 11 cases, PET upstaged eight patients (seven true positive and one false positive), and downstaged three (all false negative). In 5/65 patients, chemotherapy treatment was modified on the basis of PET findings. CONCLUSIONS: Our data confirm the high accuracy of FDG PET/CT in staging HL and NHL. FDG-PET/CT should therefore be used routinely in the initial evaluation of both patient subgroups. PMID- 18414809 TI - Role of whole-body 18F-choline PET/CT in disease detection in patients with biochemical relapse after radical treatment for prostate cancer. AB - PURPOSE: The aim of this study was to evaluate the role of whole body 18F-choline (FCH) positron emission tomography-computed tomography (PET-CT) in detecting and localising disease recurrence in patients presenting biochemical relapse after radical treatment for prostate cancer. MATERIALS AND METHODS: Fifty-six consecutive patients with increased serum prostate-specific antigen (PSA) levels after radical prostatectomy were included in the study. None of them was receiving hormone treatment at the time of the examination or had been treated during the previous 6 months. All patients underwent whole-body 18F-choline PET imaging, and the pathological findings were compared with those of further imaging exams, biopsy and follow-up. On the basis of the PSA levels, we divided our patient population into three subgroups: PSA < or = 1, 1 < PSA < or = 5, and PSA > 5 ng/ml. RESULTS: Overall, the PET scan detected disease relapse in 42.9% of cases (24/56). PET sensitivity was closely related to serum PSA levels, showing values of 20%, 44% and 81.8% in the PSA < or = 1, 1 < PSA < or = 5 and PSA > 5 ng/ml subgroups, respectively. CONCLUSIONS: In patients with biochemical relapse after radical treatment for prostate cancer, 18F-choline PET-CT represents a single step, whole-body, noninvasive study that allows disease detection and localisation. The disease detection rate is related to serum PSA levels. PMID- 18414810 TI - Lipomatous lesions of the head and neck region: imaging findings in comparison with histological type. AB - PURPOSE: Aim of the study is to demonstrate the main role of magnetic resonance imaging in the identification and characterization of lipomatous lesions of the head and neck. MATERIALS AND METHODS: CT and MRI findings of 78 patients (43 male, 35 female) aged 12-80 (mean 47.5) years surgically treated for lipomatous lesions of the head and neck region between January 1995 and June 2005 were retrospectively analysed and correlated with the histological results. RESULTS: On CT images, lipomas and fibrolipomas appeared as smooth (38/50 cases) or lobulated (12/50 cases) well-defined masses associated with moderate displacement of surrounding tissues; tumours had high signal intensity on MR T1-weighted images, with relative decreasing signal on T2-weighted images. Infiltrating lipomas appeared as expansile ill-defined masses with heterogeneous signal. Angiolipomas showed a characteristic contrast enhancement on both CT and MRI. In one case of sialolipoma, the lesion appeared markedly heterogeneous in signal. MR and CT images of Madelung's disease showed multiple symmetrical lipomatous masses involving the neck region. Intraosseous fatty lesions appeared as well-defined hypodense masses sometimes associated with cortical expansion and disruption. CONCLUSIONS: Both CT and MRI exams are useful for detecting lipomatous lesions. MRI, however, is more accurate in the evaluation of their extent and in the characterisation of uncommon lipomatous lesions of the head and neck, and intravenous administration of gadolinium better depicts the margins of the tumour and its vascularisation. PMID- 18414811 TI - Cerebral abscesses and necrotic cerebral tumours: differential diagnosis by perfusion-weighted magnetic resonance imaging. AB - PURPOSE: This study was undertaken to evaluate the usefulness of perfusion weighted imaging (PWI) in the differential diagnosis of ring-enhancing cerebral lesions, including abscesses, high-grade gliomas and metastases. MATERIALS AND METHODS: Nine cerebral abscesses (five pyogenic, four from Toxoplasma gondii), ten glioblastomas and five cerebral metastases in 19 patients were studied with gadolinium-enhanced magnetic resonance imaging, diffusion-weighted imaging (DWI) including calculation of mean apparent diffusion coefficient (ADC) of the lesion core, and PWI. At PWI, the mean of the maximum regional cerebral blood volume (rCBV) was calculated in the gadolinium-enhancing peripheral solid areas and compared with that of the contralateral normal-appearing white matter [ratio=rCBV (lesion)/rCBV (contralateral normal-appearing white matter)]. RESULTS: DWI achieved the differential diagnosis in all cases except for the four Toxoplasma abscesses. At PWI, the mean ratio of the rCBV of the capsular portion was 0.72+/ 0.08 (range 0.60-0.82) in the pyogenic abscesses, 0.84+/-0.07 (range 0.75-0.91) in the Toxoplasma abscesses, 4.45+/-1.5 (range 2.9-8.0) in the high-grade gliomas and 3.58+/-0.68 (range 3.28-4.27) in the metastases. CONCLUSIONS: PWI seems to be useful in the differential diagnosis of ring-enhancing cerebral lesions. High rCBV values in the peripheral areas appear to indicate the possibility of a necrotic tumour, whereas low values tend to indicate an abscess. PMID- 18414812 TI - What is the sensitivity of mammography and dynamic MR imaging for DCIS if the whole-breast histopathology is used as a reference standard? AB - PURPOSE: Our purpose was to compare mammography and dynamic contrast-enhanced magnetic resonance imaging (MRI) in the detection of ductal carcinoma in situ (DCIS). MATERIALS AND METHODS: Ninety patients (aged 58.6+/-16.1 years) who were candidates for unilateral (n=81) or bilateral (n=9) mastectomy underwent mammography and dynamic contrast-enhanced breast MRI using a coronal three dimensional gradient-echo sequence with slice thickness < or =3 mm before and after intravenous injection of gadoteridol (0.1 mmol/kg). Mammographic and MR images were evaluated by two offsite readers working in consensus. Pathological examination performed on 5-mm sections covering the whole breast was used as a reference standard. RESULTS: Out of 99 breasts, pathology revealed 26 DCIS in 14 breasts of 14 patients, aged 52.0 +/- 9.6 years. Lesion diameter at pathology was <5 mm (n=4); > or =5 and <10 mm (n=7); > or =10 and <20 mm (n=3); > or =20 mm (n=2); not assessed (n=10). Sensitivity was 35% (9/26) for mammography and 38% (10/26) for MRI (not significant difference, McNemar test). Both mammography and MRI provided a true positive result in seven cases (four of them measured at pathology, with a diameter of 20.0+/-12.9 mm; median 20 mm) and a false negative result in 14 cases (10 of them measured at pathology, with a diameter of 4.2+/ 1.9 mm; median 4.6 mm) (p=0.024, Mann-Whitney U test). Only 46% (12/26) of DCIS were detected at mammography and/or MRI; the remaining 54% (14/26) were diagnosed only at pathological examination. CONCLUSIONS: When the whole breast is used as the histopathological reference standard, both mammography and MRI show low sensitivity for DCIS. PMID- 18414813 TI - Phenylacetate metabolism in thermophiles: characterization of phenylacetate-CoA ligase, the initial enzyme of the hybrid pathway in Thermus thermophilus. AB - Phenylacetate-CoA ligase (E.C. 6.2.1.30), the initial enzyme in the metabolism of phenylacetate, was studied in Thermus thermophilus strain HB27. Enzymatic activity was upregulated during growth on phenylacetate or phenylalanine. The phenylacetate-CoA ligase gene (paaK) was cloned and heterologously expressed in Escherichia coli and the recombinant protein was purified. The enzyme catalyzed phenylacetate + CoA + MgATP --> phenylacetyl-CoA + AMP + MgPP(i) with a V(max) of 24 micromol/min/mg protein at a temperature optimum of 75 degrees C. The apparent K(m) values for ATP, CoA, and phenylacetate were 6, 30, and 50 microM: , respectively. The protein was highly specific toward phenylacetate and showed only low activity with 4-hydroxyphenylacetate. Despite an amino acid sequence identity of >50% with its mesophilic homologues, phenylacetate-CoA ligase was heat stable. The genome contained further homologues of genes, which are postulated to be involved in the CoA ester-dependent metabolic pathway of phenylacetate (hybrid pathway). Enzymes of this thermophile are expected to be robust and might be useful for further studies of this yet unresolved pathway. PMID- 18414814 TI - Trace element status of chronic renal patients undergoing hemodialysis. AB - The purpose of this study was to examine the status of trace elements (Cu, Zn, and Fe) and minerals (Mg, K, Na, and Cl) and the level of biochemical parameters (urea, creatinine, total protein, albumin, and glucose) in hemodialysis (HD) patients. This study included 30 HD patients (25 men and 5 women) aged 52.12 +/- 3.13 years and 30 healthy subjects (23 men and 7 women) aged 51.64 +/- 2.22 years. This study investigated the status of trace elements and minerals in HD patients. It was found that the total HD patients (before and after dialysis) had statistically lower Zn and albumin in the after-dialysis group K and Cl levels and higher Mg, creatinine, and urea in the before-dialysis group K and in the after-dialysis group glucose levels than those of the controls. It was determined that the results might be helpful in monitoring patients with renal failure in terms of insufficiency or excess of trace elements and minerals. There was positive correlation for Mg-K (r = 0.64; p = 0.001), creatinine-urea (r = 0.59; p = 0.001), K-urea (r = 0.56; p = 0.001), K-creatinine (r = 0.52; p = 0.003), Mg creatinine (r = 0.47; p = 0.008), Zn-albumin (r = 0.40; p = 0.028), and Zn creatinine (r = 0.40; p = 0.031) in the before-dialysis session. There was also positive correlation for creatinine-urea (r = 0.56; p = 0.001), K-urea (r = 0.39; p = 0.035), and Mg-creatinine (r = 0.38; p = 0.041) in the after-analysis session. As a result of the analysis of regression between serum levels of albumin and zinc in total HD patients, the use of the level of albumin might be a suitable choice in determining zinc deficiency resulting from the decrease in the level of zinc in parallel to that of albumin. The results also suggest that the relationship between creatinine and K, Mg, and Zn could be ascribed to the loss of renal function. PMID- 18414815 TI - Serum lipid levels and 3-month prognosis in Chinese patients with acute stroke. AB - INTRODUCTION: The possible correlation between serum lipid levels and outcome after stroke is still controversial. Therefore we examined whether serum lipid levels at admission had any prognostic value in the 3-month outcome after stroke. METHODS: We performed a prospective, observational study of 649 patients with acute ischaemic stroke and intracerebral haemorrhagic stroke (ICH). Information on age, sex, history of arterial hypertension, diabetes mellitus, drinking, current smoking status, stroke type, Glasgow Coma Scale and Scandinavian Stroke Scale score, time from stroke onset, and presence of atrial fibrillation was obtained. Serum lipid levels were measured in blood samples taken from fasting patients 12 to 48 hours following ictus. Death and poor neurological outcome (Modified Rankin Scale score of > or =3 points) were defined as outcome events. A logistic regression model was performed to estimate the effect of the above variables on outcome after stroke. RESULTS: We found that the median levels of serum total cholesterol (TC), triglycerides (TG) and high-density lipoprotein cholesterol (HDL-C) in good outcome patients with acute stroke were significantly higher (P<0.005) than those of poor outcome patients. The low levels of serum TC, TG and HDL-C (P<0.05) were independently related to increased 3-month poor outcome after acute ischaemic stroke and ICH. However, there was no significant relationship between LDL-C levels and 3-month outcome. CONCLUSION: The data from this study show that low levels of serum TC, TG and HDL-C are strong independent predictors of 3-month poor outcome in patients with acute ischaemic stroke and ICH. PMID- 18414816 TI - Is the use of insulin analogues cost-effective? AB - Many patients with diabetes fail to achieve their glycaemic targets despite clear evidence that glycaemic control can prevent or delay the development of costly complications in diabetes. This article describes how insulin analogues (insulins lispro, aspart, glulisine, glargine and detemir) may have a role to play in overcoming barriers to insulin acceptance and improving adherence with therapy, and examines their cost-effectiveness as determined in published studies. Cost effectiveness studies attempt to assess all the costs and all the benefits of a particular therapy. Pharmacoeconomic models, which calculate the long-term effect of interventions, show that over periods such as 10 or 35 years the higher prescription costs of analogues are offset by a lower incidence of acute hypoglycaemic events and costly, chronic complications such as nephropathy. This results in costs per 'quality-adjusted life year' that fall well within accepted limits for good value for money. Retrospective analyses of managed care databases show that higher prescription costs for analogues are offset by lower hypoglycaemia-related costs and/or inpatient claims. Relative to human insulins, analogues provide a better balance between glycaemic control and tolerability. Patients' fear of hypoglycaemia is allayed; the pen devices used with analogues facilitate insulin injection; and the pharmacokinetic characteristics lead to increased flexibility and convenience. All these factors can help increase adherence with therapy, which may in itself be cost-saving. Taken overall, these results indicate that insulin analogues are a cost-effective therapy. PMID- 18414817 TI - [Progress in diagnostics of anorectal disorders. Part I: anatomic background and clinical and neurologic procedures]. AB - Diagnostics and therapy of anorectal disorders are still questions of surgery. Exact knowledge of functional anatomy and precise clinical examination constitute the basis for the resulting therapeutic strategies. Three-dimensional endosonography and technical advances in flexible endoscopy using high-resolution chromoendoscopy and narrow-band imaging enable exact staging and diagnosis, even of malignancies in earliest stages. Furthermore new in-vivo staining methods combined with high-resolution imaging facilitate the discrimination of inflammatory and neoplastic lesions, which often lead to diagnostic difficulties in chronic inflammatory bowel disease. Developments in neurologic testing, including surface electromyography and sacral nerve stimulation, complement the diagnostic armamentarium. PMID- 18414818 TI - [Critical limb ischemia]. AB - Critical limb ischemia can be divided into acute and chronic forms. All patients suspected of the acute form should be evaluated immediately, as urgent revascularization is usually necessary. Chronic critical limb ischemia is the most complicated type of peripheral artery disease (PAD). Typical for it are pain at rest and necrosis or gangrene. The most important known risk factors are nicotine, hypertension, diabetes, and hyperlipoproteinemia. Patients suffering from PAD run a significantly higher risk of cardiovascular or cerebrovascular events and therefore should be on permanent antithrombotic therapy as secondary prophylaxis. Besides clinical examination and determination of the ankle/brachial index, complementary imaging is mandatory. Here angiography is the gold standard. For therapy surgical and other interventional procedures are available. The decision on individual therapy follows an interdisciplinary approach based on an international consensus (TASC classification). PMID- 18414820 TI - [Cup-shaped painful ulcer of abdominal wall. Diffuse dermal angiomatosis]. PMID- 18414821 TI - The association of telomere length with paternal history of premature myocardial infarction in the European Atherosclerosis Research Study II. AB - Inter-individual variability in telomere length is highly heritable and has been correlated with risk of coronary heart disease (CHD). Our aim was to determine the association of mean leukocyte telomere length with paternal history of premature myocardial infarction (MI). Mean leukocyte telomere length was measured with real-time polymerase chain reactions in 369 male students (18-28 years) with a paternal history of MI before the age of 55, recruited from 14 European universities, serving as cases and 396 age-matched controls with no paternal history of CHD. Overall, cases had borderline significantly shorter mean length (approximately 550 bp), adjusted for age and geographical region, than controls (p = 0.05). A significant difference in telomere length across the geographical regions of Europe was observed (p < 0.0001), with shorter mean length in the Baltic and South and the longest in the Middle. The case-control difference ( approximately 2.24 kb) in mean length was highly significant only in the Baltic region (p < 0.0001). There is suggestive evidence that, in young men, the biological expression of a paternal history of premature MI is at least in part mediated through inherited short telomeres. The association with paternal history of MI is strongly seen only in the Baltic compared to the rest of Europe, but this is not explained by shorter telomere length in this region. PMID- 18414822 TI - Update on tissue renin-angiotensin systems. AB - Angiotensin (Ang) II is not only generated in the circulation by renin and angiotensin-converting enzyme (ACE) but also is produced locally in numerous organs including kidney, vessels, heart, adrenal gland, eye, testis, and brain. Furthermore, widely distributed mast cells have been shown to be a production site. Local Ang II production process is commonly termed the result of a "tissue" renin-angiotensin system (RAS). Because pharmacological experiments do not easily allow targeting of specific tissues, many novel findings about the functional importance of tissue RAS have been collected from transgenic rodent models. These animals either overexpress or lack RAS components in specific tissues and thereby elucidate their local functions. The data to date show that in most tissues local RAS amplify the actions of circulating Ang II with important implications for physiology and pathophysiology of cardiovascular diseases. This review summarizes the recent findings on the importance of tissue RAS in the most relevant cardiovascular organs. PMID- 18414823 TI - [Calcaneonavicular coalition fracture. A rare differential diagnosis of post traumatic ankle pain]. AB - The causes of chronic posttraumatic ankle pain are manifold, ranging from ligament and tendon injuries and fractures to joint degeneration. Calcaneonavicular coalition, a bridge between the calcaneus and navicular bones, is a rare disease with an incidence below 1%. The morphology of this coalition is variable. It can occur as either a fibrous, cartilaginous or bony union of the involved bones. Symptoms usually occur in adolescence with chronic pain around the ankle, mainly lateral and anterior, following distortion trauma. Patients often claim to have a lateral ankle sprain. We report 2 cases of a fracture of the calcaneonavicular coalition as the cause of chronic posttraumatic ankle pain and demonstrate the characteristics of tarsal coalitions with a review of the literature. PMID- 18414824 TI - Nuptial gifts and sexual behavior in two species of spider (Araneae, Trechaleidae, Paratrechalea). AB - Male delivering of a prey packed in silk as a nuptial gift is rare in spiders and restricted until now to Pisauridae. Here, we describe this behavioral pattern found in two Trechaleidae species, Paratrechalea azul Carico 2005, and Paratrechalea ornata (Mello-Leitao 1943), mainly based on field observations. We observed the following steps of sexual behavior: sperm induction, nuptial gift construction, mate searching, pre-copulatory courtship, copulation, and copulatory ending. In this group, a nuptial gift consists of a prey wrapped in silk, which appears as a white rounded shape. The male carries his nuptial gift in his chelicerae while searching for a female. When he finds a female, he shows a stereotyped courting behavior consisting of a hyperflexed posture that is also assumed by the receptive female while they face each other. The copulatory position and pattern is similar to that found in other Lycosoidea: the male mounts the female and makes a total of four palpal insertions while alternating sides. However, after each palpal insertion the male dismounts and returns to a frontal position while biting the gift. Copulatory courtship is evidenced by palpal and leg movements. The copulation ends by female initiative and she almost always retains the nuptial gift. No case of pre-copulatory or post-copulatory cannibalism has been recorded. Descriptions of nuptial gift construction by males and copulation in these species, as well as hypotheses about the origin of nuptial gift construction among spiders, are presented. These descriptions are the first records of such nuptial gift offering for Neotropical spiders and for non-Pisauridae species as well. PMID- 18414825 TI - Pheromonal regulation of starvation resistance in honey bee workers (Apis mellifera). AB - Most animals can modulate nutrient storage pathways according to changing environmental conditions, but in honey bees nutrient storage is also modulated according to changing behavioral tasks within a colony. Specifically, bees involved in brood care (nurses) have higher lipid stores in their abdominal fat bodies than forager bees. Pheromone communication plays an important role in regulating honey bee behavior and physiology. In particular, queen mandibular pheromone (QMP) slows the transition from nursing to foraging. We tested the effects of QMP exposure on starvation resistance, lipid storage, and gene expression in the fat bodies of worker bees. We found that indeed QMP-treated bees survived much longer compared to control bees when starved and also had higher lipid levels. Expression of vitellogenin RNA, which encodes a yolk protein that is found at higher levels in nurses than foragers, was also higher in the fat bodies of QMP-treated bees. No differences were observed in expression of genes involved in insulin signaling pathways, which are associated with nutrient storage and metabolism in a variety of species; thus, other mechanisms may be involved in increasing the lipid stores. These studies demonstrate that pheromone exposure can modify nutrient storage pathways and fat body gene expression in honey bees and suggest that chemical communication and social interactions play an important role in altering metabolic pathways. PMID- 18414826 TI - [Expanding therapeutic reference ranges using dose-related reference ranges]. AB - Evidence-based therapeutic drug monitoring (TDM), which may be successfully employed to guide drug therapy in clinical routine, supplies all the information from laboratory determination of a drug concentration in a patient's blood specimen. This value is interpreted first of all in relation to a therapeutic reference range that must be established according to the same rules that are generally accepted for clinical studies aimed to license a new drug. The drug concentration may be furthermore interpreted in reference to a dose-related reference range. Thereby a signal is created to alert for individual abnormalities such as drug/drug interactions, gene polymorphisms that give rise to slow/rapid metabolizers, altered function of the excretion organs liver and kidneys by age and/or disease, compliance problems, a missing pharmacokinetic steady state, and even signal overlay in the laboratory analysis. We return all information available and clinical pharmacological comments to physicians who send specimens to our laboratory. PMID- 18414827 TI - [Cervical spine injury. Diagnosis, prognosis and management]. AB - Cervical spine injuries are a common occurrence in multi-trauma patients and should be taken into account when planning further clinical management. This review describes in detail upper and lower cervical spine injuries and introduces the sub-axial injury classification (SLIC) that is based on three components: injury morphology, integrity of the discoligamentous complex and the neurologic status of the patient. If the total SLIC score is <3, non-surgical treatment is recommended. If the total is > or =5, operative treatment is indicated as such a score is associated with instability and probably neurologic deficits. The precise diagnosis of the radiologist, which would include the SLIC classification, should facilitate clinical decision-making about further management. PMID- 18414828 TI - [Evidence-based diagnosis of abdominal trauma]. AB - Abdominal trauma is a common cause of death particularly in patients up to 40 years of age. In order to reduce mortality a rapid radiologic diagnosis is essential. At present, sonography plays a role only in the evaluation of minor trauma and as a "focused assessment with sonography for trauma" (FAST) to clarify free intraperitoneal fluid immediately on admittance in severely injured patients. However, computed tomography has proven to be a potent tool for the triage of patients with abdominal trauma, because, based on the results of the CT scan, patients can be referred for laparotomy or safely classified for "wait and see" treatment. Therapeutic decisions are largely based on injury severity scores and the radiologist must be familiar with them as well as with the associated therapeutic consequences. PMID- 18414829 TI - Molecular mapping of QTLs for resistance to the greenbug Schizaphis graminum (Rondani) in Sorghum bicolor (Moench). AB - Sorghum is a worldwide important cereal crop and widely cultivated for grain and forage production. Greenbug, Schizaphis graminum (Rondani) is one of the major insect pests of sorghum and can cause serious damage to sorghum plants, particularly in the US Great Plains. Identification of chromosomal regions responsible for greenbug resistance will facilitate both map-based cloning and marker-assisted breeding. Thus, a mapping experiment was conducted to dissect sorghum genetic resistance to greenbug biotype I into genomic regions. Two hundred and seventy-seven (277) F(2) progeny and their F(2:3) families from a cross between Westland A line (susceptible parent) and PI550610 (resistant parent) combined with 118 polymorphic simple sequence repeat (SSR) markers were used to map the greenbug resistance QTLs. Composite interval mapping (CIM) and multiple interval mapping (MIM) revealed two QTLs on sorghum chromosome nine (SBI 09) consistently conditioned the resistance of host plant to the greenbug. The two QTLs were designated as QSsgr-09-01 (major QTL) and QSsgr-09-02 (minor QTL), accounting for approximately 55-80%, and 1-6% of the phenotypic variation for the resistance to greenbug feeding, respectively. These resistance QTLs appeared to have additive and partially dominant effects. The markers Xtxp358, Xtxp289, Xtxp67 and Xtxp230 closely flanked the respective QTLs, and can be used in high throughput marker-assisted selections (MAS) for breeding new resistant parents and producing commercial hybrids. PMID- 18414830 TI - Human retinal pigment epithelium proteome changes in early diabetes. AB - AIMS/HYPOTHESIS: Diabetic retinopathy is the most common complication of diabetes and a leading cause of blindness among working-age adults. Anatomical and functional changes occur in the retina and retinal pigment epithelium (RPE) prior to clinical symptoms of the disease. However, the molecular mechanisms responsible for these early changes, particularly in the RPE, remain unclear. To begin defining the molecular changes associated with pre-retinopathic diabetes, we conducted a comparative proteomics study of human donor RPE. METHODS: The RPE was dissected from diabetic human donor eyes with no clinically apparent diabetic retinopathy (n=6) and from eyes of age-matched control donors (n=17). Soluble proteins were separated based upon their mass and charge using two-dimensional (2 D) gel electrophoresis. Protein spots were visualised with a fluorescent dye and spot densities were compared between diabetic and control gels. Proteins from spots with significant disease-related changes in density were identified using mass spectrometry. RESULTS: Analysis of 325 spots on 2-D gels identified 31 spots that were either up- or downregulated relative to those from age-matched control donors. The protein identity of 18 spots was determined by mass spectrometry. A majority of altered proteins belonged to two major functional groups, metabolism and chaperones, while other affected categories included protein degradation, synthesis and transport, oxidoreductases, cytoskeletal structure and retinoid metabolism. CONCLUSIONS/INTERPRETATION: Changes identified in the RPE proteome of pre-retinopathic diabetic donor eyes compared with age-matched controls suggest specific cellular alterations that may contribute to diabetic retinopathy. Defining the pre-retinopathic changes affecting the RPE could provide important insight into the molecular events that lead to this disease. PMID- 18414831 TI - Hyphenated mass spectrometry in the analysis of the central carbon metabolism. PMID- 18414832 TI - Fast determination of toxic diethylene glycol in toothpaste by ultra-performance liquid chromatography-time of flight mass spectrometry. AB - A rapid method for determining diethylene glycol (DEG) in toothpaste based on the use of ultra-performance liquid chromatography (UPLC) coupled to time-of-flight mass spectrometry (TOF-MS) has been developed. The method has been validated in toothpaste samples spiked at different levels, 0.005, 0.1 and 5%, obtaining satisfactory recoveries (74-98%) and relative standard deviations (<4%). Quantification was carried out by using matrix-matched standards calibration. The developed method was applied to several types of toothpaste, making identification and quantification of DEG and other polyethylene glycols (PEG) feasible with very little sample manipulation, as only extraction with water is required. The excellent sensitivity of TOF-MS analysis performed in full-scan acquisition mode allowed the determination of DEG at concentration levels as low as 0.005% in samples and its reliable identification via the mass accuracy measurements provided by this instrument (<5 ppm). PMID- 18414833 TI - Analysis of aged sulfadiazine residues in soils using microwave extraction and liquid chromatography tandem mass spectrometry. AB - An efficient extraction of sulfadiazine residues from soils is difficult, as sulfadiazine is known to form quickly sequestering residues. The objective of this study was to optimize an exhaustive extraction for aged residues of sulfadiazine and its two major metabolites, N-acetylsulfadiazine and 4 hydroxysulfadiazine, from soil. For this purpose two representative used agricultural soils (Luvisol, Cambisol) were blended with manure derived from [(14)C]sulfadiazine-treated pigs and incubated at 10 degrees C in the laboratory. After different extraction tests with various solvent mixtures (two- to four component mixtures with water, methanol, acetonitrile, acetone, and/or ethyl acetate), different pH values (pH 4 and 9), and extraction temperatures (up to 200 degrees C), soil extracts were measured by liquid scintillation counting and liquid chromatography coupled to tandem mass spectrometry. With respect to sulfadiazine yields, stability of soil extracts, and the amount of coextracted matrix, a microwave extraction of soil (15 min, 150 degrees C) using acetonitrile/water 1:4 (v/v) is the method of choice for the exhaustive extraction of aged sulfadiazine residues from soils. PMID- 18414834 TI - Development and certification of the new NIES CRM 28: urban aerosols for the determination of multielements. AB - A new environmental certified reference material (CRM) for the determination of multielements in aerosol particulate matter has been developed and certified by the National Institute for Environmental Studies (NIES), Japan, based on analyses by a network of laboratories using a wide range of methods. The origin of the material was atmospheric particulate matter collected on filters in a central ventilating system in a building in Beijing city centre. The homogeneity and stability of this material were sufficient for its use as a reference material. Values for elemental mass fractions in the material were statistically determined based on the analytical results of the participating laboratories. Eighteen certified values and 14 reference values were obtained. The diameters, obtained from a micrographic image using image analysis software, of 99% of the particles were less than 10 microm, demonstrating that almost all the particles in the material could be classified as particles of 10 microm or less in aerodynamic diameter. The chemical composition and particle size distribution of this material were close to those of an authentic aerosol collected in Beijing. NIES CRM 28 is appropriate for use in analytical quality control and in the evaluation of methods used in the analysis of aerosols, particularly those collected in urban environments in northeast Asia. PMID- 18414836 TI - Sensitive light-scattering detection-magnetophoretic acceleration mass analysis of single microparticles in an atmosphere. AB - Optically detected magnetophoretic acceleration mass analysis of an individual micro-particle in an atmosphere has been remarkably improved in sensitivity by using a reflective microscope objective, by which forward scattered light from a particle could be effectively collected. From the light-scattering simulation, the detection limit for the radius of a micro-particle was estimated to be smaller than 0.4 MUm, and about 60 times intensity enhancement was observed for a polystyrene particle with a radius of 2.8 MUm. For both paramagnetic and diamagnetic micro-particles, the mass magnetic susceptibility and the relaxation time could be determined without knowing any parameters of the particles. From the relaxation time, the mass of a particle was obtained if the radius or the density of the particle was known. For a test sample silica particles were used to adsorb paramagnetic dysprosium(III), the surface concentration of dysprosium(III) on a single particle could be successfully determined by use of this method. PMID- 18414835 TI - Simultaneous speciation analysis of glutathione peroxidase, selenoprotein P and selenoalbumin in human serum by tandem anion exchange-affinity HPLC and on-line isotope dilution ICP-quadrupole MS. AB - A method based on anion exchange (AE) and affinity (AF)-HPLC (AE-AF-HPLC) hyphenated to inductively coupled plasma-(quadrupole) mass spectrometry (ICP-QMS) was developed for the speciation analysis of selenoprotein P (SelP), glutathione peroxidase (GPx) and selenoalbumin (SeAlb) in human serum. AE-HPLC is proposed here for the on-line alleviation of Cl and Br spectral interferences on (77)Se ((40)Ar(37)Cl) and (82)Se ((81)Br(1)H). Separation of GPx, SelP and SeAlb by AE AF-HPLC was obtained within a total chromatographic runtime of <20 min. On-line (post-column) isotope dilution (ON-ID) and on-line external calibration (ON-EC) ICP-QMS were used for the quantification of Se in GPx, SelP and SeAlb. ON-EC using a Se-L: -cystine standard was shown to be a suitable approach for the routine simultaneous speciation analysis of serum GPx, SelP and SeAlb. The method validation was carried out by direct ICP-sector field MS determination of Se in GPx, SelP and SeAlb fractions collected after AE-AF-HPLC separation. In addition, the method accuracy for the determination of total protein-bound Se was assessed by analyzing a human serum reference material (BCR-637) certified for total Se content. PMID- 18414837 TI - The use of electrochemical impedance spectroscopy for biosensing. AB - This review introduces the basic concepts and terms associated with impedance and techniques of measuring impedance. The focus of this review is on the application of this transduction method for sensing purposes. Examples of its use in combination with enzymes, antibodies, DNA and with cells will be described. Important fields of application include immune and nucleic acid analysis. Special attention is devoted to the various electrode design and amplification schemes developed for sensitivity enhancement. Electrolyte insulator semiconductor (EIS) structures will be treated separately. PMID- 18414838 TI - Validation of hippocampal volumes measured using a manual method and two automated methods (FreeSurfer and IBASPM) in chronic major depressive disorder. AB - INTRODUCTION: To validate the usefulness of the packages available for automated hippocampal volumetry, we measured hippocampal volumes using one manual and two recently developed automated volumetric methods. METHODS: The study included T1 weighted magnetic resonance imaging (MRI) of 21 patients with chronic major depressive disorder (MDD) and 20 normal controls. Using coronal turbo field echo (TFE) MRI with a slice thickness of 1.3 mm, the hippocampal volumes were measured using three methods: manual volumetry, surface-based parcellation using FreeSurfer, and individual atlas-based volumetry using IBASPM. In addition, the intracranial cavity volume (ICV) was measured manually. RESULTS: The absolute left hippocampal volume of the patients with MDD measured using all three methods was significantly smaller than the left hippocampal volume of the normal controls (manual P = 0.029, FreeSurfer P = 0.035, IBASPM P = 0.018). After controlling for the ICV, except for the right hippocampal volume measured using FreeSurfer, both measured hippocampal volumes of the patients with MDD were significantly smaller than the measured hippocampal volumes of the normal controls (right manual P = 0.019, IBASPM P = 0.012; left manual P = 0.003, FreeSurfer P = 0.010, IBASPM P = 0.002),. In the intrarater reliability test, the intraclass correlation coefficients (ICCs) were all excellent (manual right 0.947, left 0.934; FreeSurfer right 1.000, left 1.000; IBASPM right 1.000, left 1.000). In the test of agreement between the volumetric methods, the ICCs were right 0.846 and left 0.848 (manual and FreeSurfer), and right 0.654 and left 0.717 (manual and IBASPM). CONCLUSION: The automated hippocampal volumetric methods showed good agreement with manual hippocampal volumetry, but the volume measured using FreeSurfer was 35% larger and the agreement was questionable with IBASPM. Although the automated methods could detect hippocampal atrophy in the patients with MDD, the results indicate that manual hippocampal volumetry is still the gold standard, while the automated volumetric methods need to be improved. PMID- 18414840 TI - Inadvertent intraosseous gadolinium injection during pediatric shoulder MR arthrography. AB - A variety of problems can arise while performing MR arthrography of the shoulder. These have been well documented in the literature and range from improper patient selection to errors in technique or in the choice of imaging sequences. We present a rare case of inadvertent, painless intraosseous injection of dilute gadolinium into the proximal humeral epiphysis of a 13-year-old male athlete. The clinical relevance is discussed and technical recommendations are offered. PMID- 18414839 TI - The pathogenesis and imaging of the tuberous sclerosis complex. AB - Tuberous sclerosis complex (TSC) is an autosomal-dominant disorder characterized by the formation of hamartomatous lesions in multiple organ systems. It is the second most common neurocutaneous syndrome after neurofibromatosis type 1 and has been recognized since the late 1800s. Although the disease has complete penetrance, there is also high phenotypic variability: some patients have obvious signs at birth, while others remain undiagnosed for many years. In addition to skin lesions, TSC patients develop numerous brain lesions, angiomyolipoma (AMLs), lymphangiomyomatosis (LAM) in the lungs, cardiac rhabdomyomas, skeletal lesions, and vascular anomalies, all of which are well seen with medical imaging. Our knowledge of TSC genetics and pathophysiology has expanded dramatically in recent years: two genetic loci were discovered in the 1990s and recent elucidation of TSC's interaction with the mTOR pathway has changed how we manage the disease. Meanwhile, medical imaging is playing an increasingly important role in the diagnosis, management, and treatment of TSC. We provide an update on the genetics and pathophysiology of TSC, review its clinical manifestations, and explore the breadth of imaging features in each organ system, from prenatal detection of cardiac rhabdomyomas to monitoring rapamycin therapy to treatment of AMLs by interventional radiology. PMID- 18414842 TI - Duplicate publication with consequence of potential misinformation and further controversy. PMID- 18414841 TI - Efficacy of a radiation safety education initiative in reducing radiation exposure in the pediatric IR suite. AB - BACKGROUND: The use of ionizing radiation is essential for diagnostic and therapeutic imaging in the interventional radiology (IR) suite. As the complexity of procedures increases, radiation exposure risk increases. We believed that reinforcing staff education and awareness would help optimize radiation safety. OBJECTIVE: To evaluate the effect of a radiation safety education initiative on IR staff radiation safety practices and patient radiation exposure. MATERIALS AND METHODS: After each fluoroscopic procedure performed in the IR suite during a 4 month period, dose-area product (DAP), fluoroscopy time, and use of shielding equipment (leaded eyeglasses and hanging lead shield) by IR physicians were recorded. A lecture and article were then given to IR physicians and technologists that reviewed ALARA principles for optimizing radiation dose. During the following 4 months, those same parameters were recorded after each procedure. RESULTS: Before education 432 procedures were performed and after education 616 procedures were performed. Physician use of leaded eyeglasses and hanging shield increased significantly after education. DAP and fluoroscopy time decreased significantly for uncomplicated peripherally inserted central catheters (PICC) procedures and non-PICC procedures after education, but did not change for complicated PICC procedures. CONCLUSION: Staff radiation safety education can improve IR radiation safety practices and thus decrease exposure to radiation of both staff and patients. PMID- 18414843 TI - Migration of Gelfoam to the gallbladder after liver biopsy. AB - Liver biopsy is a common procedure, with an inherent risk of bleeding. There are different ways to help avoid hemorrhage, including biopsy through a transjugular venous route or embolization of the tract with liquid or solid materials. We describe an image-guided percutaneous core needle liver biopsy with tract embolization using thick Gelfoam slurry in a pediatric oncology patient. Imaging studies acquired after the biopsy indicated that the Gelfoam mixture had likely migrated to the gallbladder and common bile duct. We report this rare occurrence with its striking imaging in order to make those performing biopsies aware of this possibility. PMID- 18414845 TI - Selectivity in the mechanism of action of antimicrobial mastoparan peptide Polybia-MP1. AB - Many potent antimicrobial peptides also present hemolytic activity, an undesired collateral effect for the therapeutic application. Unlike other mastoparan peptides, Polybia-MP1 (IDWKKLLDAAKQIL), obtained from the venom of the social wasp Polybia paulista, is highly selective of bacterial cells. The study of its mechanism of action demonstrated that it permeates vesicles at a greater rate of leakage on the anionic over the zwitterionic, impaired by the presence of cholesterol or cardiolipin; its lytic activity is characterized by a threshold peptide to lipid molar ratio that depends on the phospholipid composition of the vesicles. At these particular threshold concentrations, the apparent average pore number is distinctive between anionic and zwitterionic vesicles, suggesting that pores are similarly formed depending on the ionic character of the bilayer. To prospect the molecular reasons for the strengthened selectivity in Polybia-MP1 and its absence in Mastoparan-X, MD simulations were carried out. Both peptides presented amphipathic alpha-helical structures, as previously observed in Circular Dichroism spectra, with important differences in the extension and stability of the helix; their backbone solvation analysis also indicate a different profile, suggesting that the selectivity of Polybia-MP1 is a consequence of the distribution of the charged and polar residues along the peptide helix, and on how the solvent molecules orient themselves according to these electrostatic interactions. We suggest that the lack of hemolytic activity of Polybia-MP1 is due to the presence and position of Asp residues that enable the equilibrium of electrostatic interactions and favor the preference for the more hydrophilic environment. PMID- 18414846 TI - Identification of fungal metabolites of anticonvulsant drug carbamazepine. AB - Carbamazepine, which has been used in the treatments of epilepsy, is often found in the environment. Although metabolism of carbamazepine by humans and rats has been characterized, the environmental fate of carbamazepine has not been studied. In this study, two model fungi Cunninghamella elegans ATCC 9245 and Umbelopsis ramanniana R-56, which have previously shown diverse metabolic activities, were tested for metabolism of carbamazepine. Both fungi produced three metabolites each (C1-C3 and M1-M3). All six metabolites showed [M + H](+) at m/z 253, suggesting addition of one oxygen to the parent compound. High-performance liquid chromatography and liquid chromatography-mass spectrometric analysis detected 10, 11-dihydro-10, 11-epoxycarbamazepine as a major product (C3 (47%) and M3 (85%)) and 3-hydroxycarbamazepine (C2 (15%) and M2 (7%)) from carbamazepine through mixed mono-oxidation reactions in both fungal strains. C. elegans was confirmed to produce 2-hydroxycarbamazepine (C1 (38%)) while U. ramanniana produced a yet unidentified ring-hydroxylated metabolite (M1 (8%)). The current study suggests that carbamazepine is likely to be subjected to initially diverse mono oxygenation reactions by fungal metabolisms, resulting in the formation of the corresponding metabolites, which were similarly found in mammalian metabolisms. PMID- 18414844 TI - Inhibition of P2X(7) receptors by divalent cations: old action and new insight. AB - P2X family receptors form ATP-gated ion channels by assembling homo/hetero trimers from seven receptor subunits. The homomeric P2X(7) receptor is extraordinary in that in addition to distinctive localization and biological functions it exhibits several hallmark properties, for example, the receptor is potently inhibited by divalent cations such as calcium, magnesium, zinc and copper. Despite the fact that this distinct feature was first described almost three decades ago, our understanding is still contentious. Recent site-directed mutagenesis studies have provided direct evidence showing that functional inhibition by zinc and copper primarily results from direct interaction with the receptor. In this short review, I will give a concise description of the major localization, biological functions, and unique properties of the P2X(7) receptor, and particularly discuss the evolving understanding of how divalent cations inhibit the P2X(7) receptor and the potential implication of such inhibition to the physiological and pathophysiological role of the P2X(7) receptor. PMID- 18414847 TI - Intergeneric coaggregation of strains isolated from phenol-degrading aerobic granules. AB - This work aims at exploring the intergeneric coaggregation of the pairs of strains, Acinetobacter calcoaceticus I6 and Bacillus thuringiensis I2 or Candida tropicalis I9 (with GenBank accession numbers EU250016, EU036759, and DQ515822) isolated from phenol-degrading aerobic granules. The I2 and I6 are functionally similar stains, while the I6 and I9 are functionally dissimilar strains. The lectin-saccharide interaction controlled the coaggregation of both the I2+I6 and I6+I9 pairs, with the protein adhesin being associated with the strain I6, and the complementary galactosamine-like or fucose-like sugar receptor with the strain I2 or I9, respectively. The rod-like I2 cells bridged the clusters of I2 or I6 cells to form aggregates, while the small I6 cells attached on and modified the surface of I9 to form aggregates. PMID- 18414848 TI - Cloning, characterization and functional expression of an alkalitolerant type C feruloyl esterase from Fusarium oxysporum. AB - A hypothetical protein FoFaeC-12213 of Fusarium oxysporum was found to have high amino acid sequence identity with known type C feruloyl esterases (FAEs) containing a 13-amino acid conserved region flanking the characteristic G-X-S-X-G motif of a serine esterase. The putative FAE from the genomic DNA was successfully cloned in frame with the Saccharomyces cerevisiae alpha-factor secretion signal under the transcriptional control of the alcohol oxidase (AOX1) promoter and integrated in Pichia pastoris X-33 to confirm that the enzyme exhibits FAE activity. The molecular weight (62 kDa) and pI (6.8) were in agreement with the theoretical calculated values indicating the correct processing of the secretion signal in P. pastoris. The recombinant FAE was purified to its homogeneity and subsequently characterized using a series of model substrates including methyl esters of hydroxycinnamates, alkyl ferulates and monoferuloylated 4-nitrophenyl glycosides. The substrate specificity profiling reveals that the enzyme is a type C FAE showing broad hydrolytic activity against the four methyl esters of hydroxycinnamic acids and strong preference for the hydrolysis of n-propyl ferulate. Ferulic acid (FA) was efficiently released from destarched wheat bran when the esterase was incubated together with xylanase from Trichoderma longibrachiatum (a maximum of 67% total FA released after 1-h incubation). The esterase showed broad pH stability making it an important candidate for alkaline applications such as pulp treatment in the paper industry. PMID- 18414849 TI - Investigating the interaction mechanism between zinc and Saccharomyces cerevisiae using combined SEM-EDX and XAFS. AB - The interaction mechanism between zinc and the intact yeast cells of Saccharomyces cerevisiae was investigated by using the scanning electron microscopy with energy-dispersive X-ray analysis, as well as X-ray absorption fine structure spectroscopy (XAFS). Displacement of H+, K+, Mg2+, and Na+ during zinc uptake confirmed the existence of both covalent interactions and ionic interactions between Zn2+ and the microbe. Ion exchange mechanism played a role in zinc uptake. The local environment of Zn accumulated in the intact yeast cells was determined by XAFS, which suggests that the nearest neighboring atom of the bound zinc ion on the biomass is oxygen atom. The adsorbed zinc ion on the intact cells of S. cerevisiae is a tetrahedron structure, with the Zn-O bond length of 1.97 A, and the coordination number is only 3.2 of Zn-O structure in the first shell. PMID- 18414850 TI - Lovastatin biosynthetic genes of Aspergillus terreus are expressed differentially in solid-state and in liquid submerged fermentation. AB - Molecular studies were performed to establish the causes of the superior lovastatin productivity of a novel solid-state fermentation (SSF) process, in relation with liquid submerged fermentation (SmF; 20 mg/g vs. 0.65 mg/ml). In SSF, biosynthetic genes lovE and lovF transcripts accumulated to high levels from day 1 to day 7. In this period, lovE transcript showed 4.6-fold higher accumulation levels (transcription) than the highest level detected in SmF (day 5). lovF transcript showed two-fold higher expression than the highest point in SmF. In SmF, the expression was only detected clearly on day 5 and, showing a 50% decrease, on day 7. These results show that the higher lovastatin production in SSF is related to a more intense transcription of these biosynthetic genes. A strong expression of gldB gene in lovastatin SSF indicated that Aspergillus terreus senses osmotic stress during the course of SSF, but not in SmF. However, when a liquid medium of identical concentration was used in SmF, lovastatin production decreased in SSF. PMID- 18414851 TI - Hepatitis C-associated osteosclerosis: a case report. AB - A 38-year-old man presented for evaluation of severe pain in his lower limbs of 16 months' duration. Radiographs showed diffuse bony sclerosis involving the axial and appendicular skeleton with marked cortical thickening in the diaphyseal regions of the long bones. Iliac crest biopsy revealed abundant sclerotic bone. The patient was diagnosed with hepatitis C secondary to intravenous drug use 20 years earlier, and the bony findings and clinical symptoms attributed to hepatitis C-associated osteosclerosis (HCAO). Little is known about the natural history of this disease; however, we discuss the patient's clinical course over 15 years. PMID- 18414852 TI - Of impact, metrics and ethics. PMID- 18414853 TI - Pre-treatment neutrophil to lymphocyte ratio is elevated in epithelial ovarian cancer and predicts survival after treatment. AB - PURPOSE: Inflammatory cells can both suppress and stimulate tumor growth, and the influence of inflammatory cells on clinical outcome has been the focus of many studies. The purpose of this study was to evaluate the effectiveness of the neutrophil to lymphocyte ratio (NLR), a measure of the systemic inflammatory response, as an additional discriminative biomarker in epithelial ovarian cancer and to determine whether it predicts survival and recurrence. METHODS: We studied 192 patients with epithelial ovarian cancer, 173 with benign ovarian tumors, 229 with benign gynecologic disease, and 405 healthy controls. Serum CA125 levels and leukocyte counts according to subtypes were recorded prior to treatment in all study subjects. In epithelial ovarian cancer, the diagnostic usefulness of NLR, in combination with CA125, was evaluated. The correlation between NLR and overall and disease-free survival was analyzed using both univariate and multivariate analyses adjusting for the known prognostic factors (age, stage, cell type, and grade). RESULTS: Preoperative NLR in ovarian cancer subjects (mean 6.02) was significantly higher than that in benign ovarian tumor subjects (mean 2.57), benign gynecologic disease subjects (mean 2.55), and healthy controls (mean 1.98) (P < 0.001). The sensitivity and specificity of NLR in detecting ovarian cancer was 66.1% (95% CI, 59.52-72.68%) and 82.7% (95% CI, 79.02-86.38%), respectively (cutoff value: 2.60). In early stage ovarian cancer, CA125 was not elevated in 19 out of 49 patients. Seven (36.8%) of these 19 patients were NLR positive. On Cox multivariate analysis, NLR positive, stage III/IV, and older age were independent poor prognostic factors, and being NLR positive was the most powerful predictive variable (Hazard Ratio = 8.42 [95% CI: 1.09-64.84], P = 0.041). CONCLUSIONS: Our findings provide evidence for the association between NLR and epithelial ovarian cancer. Preoperative NLR, in combination with CA125, may represent a simple and cost-effective method of identifying ovarian cancers, and an elevated NLR may predict an adverse outcome in ovarian cancer. PMID- 18414854 TI - Selective accumulation of differentiated FOXP3(+) CD4 (+) T cells in metastatic tumor lesions from melanoma patients compared to peripheral blood. AB - Precise identification of regulatory T cells is crucial in the understanding of their role in human cancers. Here, we analyzed the frequency and phenotype of regulatory T cells (Tregs), in both healthy donors and melanoma patients, based on the expression of the transcription factor FOXP3, which, to date, is the most reliable marker for Tregs, at least in mice. We observed that FOXP3 expression is not confined to human CD25(+/high) CD4(+) T cells, and that these cells are not homogenously FOXP3(+). The circulating relative levels of FOXP3(+) CD4(+) T cells may fluctuate close to 2-fold over a short period of observation and are significantly higher in women than in men. Further, we showed that FOXP3(+) CD4(+) T cells are over-represented in peripheral blood of melanoma patients, as compared to healthy donors, and that they are even more enriched in tumor infiltrated lymph nodes and at tumor sites, but not in normal lymph nodes. Interestingly, in melanoma patients, a significantly higher proportion of functional, antigen-experienced FOXP3(+) CD4(+) T was observed at tumor sites, compared to peripheral blood. Together, our data suggest that local accumulation and differentiation of Tregs is, at least in part, tumor-driven, and illustrate a reliable combination of markers for their monitoring in various clinical settings. PMID- 18414855 TI - Normative data on hand grip strength in a Greek adult population. AB - The objectives of this study were to establish data concerning normal hand grip strength (GS) and to explore possible associations with anthropometric parameters. GS was measured in 232 individuals in a standard arm position using the Jamar dynamometer. We examined differences between right/left and dominant/nondominant hands. Possible correlations of GS with anthropometric values were evaluated. Right hand and dominant hand GS were found to be higher and statistically significant compared to left hand and nondominant hand GS, respectively. Men had higher values of GS compared to women. A negative association was observed between age and dominant hand GS. A positive association was documented between height and dominant hand GS, while the respective comparison for weight and dominant hand GS documented a statistically significant positive association only in the male group. A positive association between BMI and dominant hand GS was seen in female individuals. Additional factors associated with GS should be the goal of future investigations. PMID- 18414856 TI - Arthroscopic inspection after the surgical treatment of patella fractures. AB - After the treatment of patella fractures the only way to evaluate healing at the articular surface before implant removal is through arthroscopy. The purpose of this study was to examine the healing potential of the cartilage. Arthroscopy was performed in 18 patients at the time of implant removal. The mean age of the patients was 42.1 years. The time elapsed from the index surgery to the arthroscopy and implant removal surgery was 12.9 months. During the arthroscopy, we inspected articular step-off, cartilage loss, and joint surface irregularities. Cartilage irregularities were observed in 13 of the 18 patients. Five patients had well-healed cartilage at the patellar surface. Although none of the patients had displacement at final follow-up X-rays, step-off was detected in two patients during arthroscopy. Our observation showed that cartilage lesions did not correlate with clinical and radiological evaluation. Despite good knee scores, we observed surface irregularities, chondral lesions, and fibrillation in most of the cases implicating subsequent patellofemoral arthritis. PMID- 18414857 TI - Management of trochanteric fractures of the femur with external fixation in high risk patients. AB - The aim of this prospective study was to assess the outcome of trochanteric fractures of the femur after external fixation in a group of elderly patients with high surgical risk. The study population consisted of 50 patients with trochanteric fractures of the femur and a mean age of 87 years who were classified by an anaesthetist as ASA 3 or 4 and considered not suitable for conventional fractures fixation. The fracture was fixed with an external fixator under spinal anaesthesia. The final follow-up was at 12 months. All fractures healed within 12 weeks. Superficial pin tract infection occurred in 30 patients, and fracture united with a shortening of 14 mm (5-20) in 12 patients. No implant failures or limitation of knee movements were recorded. Five patients died within 1 year. External fixation is a valuable treatment alternative for trochanteric fracture of the femur in elderly patients. PMID- 18414859 TI - The influence of femoral head shift on hip biomechanics: additional parameters accounted. AB - Pauwels' method of hip biomechanics can explain a negative influence of the lateral shift of the femoral head on the load of dislocated hip joint, but, the influence of the cranial shift of femoral head can not be explained. A calculation of hip balance which takes into consideration both lateral and cranial shifts of the femoral head is presented. Two pelvic radiographs were used; of an adult person and of a two year old child. One hip was normal, and other was dislocated. Force R was established using horizontal lever k(1), and a new vertical lever k(2). Graphically and mathematically the results show that the force R is always greater in the dislocated hip. Both lateral and cranial shifts contribute to this. The modification of Pauwels method described clearly demonstrates that not only lateral but also cranial shift of the femoral head in dislocated hip should be taken into consideration. PMID- 18414858 TI - Concomitant noncontiguous level (thoracic & lumbar) spinal stenosis. AB - Presented here is a prospective study assessing the efficacy of decompression of concomitant noncontiguous level (thoracic & lumbar) stenosis in accordance with neurological findings, nerve root blocks, and myelographically proven disease. The objective was to determine the efficacy, clinical outcome, and functional recovery in patients undergoing simultaneous decompression. No previous study has focussed on the clinical outcome of such simultaneous decompression. Twenty-one patients with neurological claudication, progressive gait disturbance, upper motor neuron symptoms, and findings of myelopathy in both the lower extremities underwent simultaneous decompression and were assessed. The average follow-up was 32 months (range, 24-40 months). At the last examination,13 patients (82%) had excellent or good clinical results. Postoperative improvement correlated inversely with the duration of symptoms. The patients usually had satisfactory outcomes when the correct diagnosis was made and management was implemented. Functional recovery depends on early diagnosis and timely surgical intervention. PMID- 18414860 TI - Time and cost savings in arthroscopic subacromial decompression: the use of bipolar versus monopolar radiofrequency. AB - There is currently an increased interest in the use of electro surgery in arthroscopy. Since the introduction of the bipolar arthroscopic radiofrequency (RF) wand, it has started to replace the classic Bovie monopolar probe on the assumption that the new technology provides multifunctional devices, combining both tissue removal and haemostasis into one instrument. The more efficient tissue ablation and precise haemostasis achieved with these instruments should result in a significant reduction in the operative time and cost. We ran a prospective comparative randomised study to test this hypothesis. Forty patients underwent arthroscopic subacromial decompression, randomised into two groups. The group treated with bipolar RF was associated with an average operative time saving of 8 min (P < 0.0001) and an average cost saving of pound 83 (euro 111) per case (P < 0.003), compared to monopolar RF. Bipolar RF is the instrument of choice in arthroscopic shoulder surgery, as it saves time and money. PMID- 18414861 TI - The epidemiology of traumatic humeral shaft fractures in Taiwan. AB - We retrospectively analysed 106 consecutive traumatic humeral shaft fractures over a five-year period. The mechanism of injury, age, gender, fracture types, associated injury and the presence of injury to the radial nerve were reviewed. The incidence was about 10 per 100,000 per year; most were closed fractures in young males which had been sustained as a result of traffic accidents. The age gender distribution was characterised by gradually increased incidence from the fifth decade in women, while it reached a peak at the third decade and decreased after the fifth decade in men. The results revealed different epidemiological features from previous studies. The epidemiology differs between ethnicity and country, and updating the epidemiological features of humeral shaft fractures may provide information for appropriate treatment programmes. This study documents the epidemiology of humeral shaft fracture in Taiwan, probably for the first time in this Asian community. PMID- 18414862 TI - Thalassemia and hemoglobinopathies in pregnant Lao women: carrier screening, prevalence and molecular basis. AB - To provide relevant evidence base for implementation of a prevention and control program for thalassemia in the Lao People's Democratic Republic (Lao PDR), we have evaluated a simple screening protocol and examined the prevalence and the molecular basis of thalassemia in pregnant Lao women. The study was conducted on 307 pregnant women attending the Mother and Child Health Hospital, Vientiane. Initial screening was performed locally, applying a combined osmotic fragility (OF) and dichlorophenolindophenol (DCIP) test. Erythrocyte counts were recorded. The remaining blood specimens were transferred to Thailand for further hemoglobin (Hb) and DNA analyses. Subjects were divided into four groups according to the results of the screening tests. Among 307 participants examined, 154 (50.2%) had negative results on both tests (-/-), 58 (18.8%) were positive on the OF test but not the DCIP test (+/-), 22 (7.1%) were negative on the OF test but positive on the DCIP test (-/+), and 73 (23.7%) were positive on both tests (+/+). As many as 25 thalassemia genotypes including various complex syndromes were observed. Three clinically important forms of thalassemia including alpha(o)-, beta-thalassemia, and Hb E were identified in 39 (12.7%), 11 (3.6%), and 93 (30.2%) subjects, respectively. The performance characteristic of the initial screening for these three forms of thalassemia was determined. The sensitivity, specificity, positive, and negative predictive values were found to be 99.2%, 85.5%, 83.0% and 99.4%, respectively. Therefore, thalassemia and hemoglobinopathies are prevalent and heterogeneous among the Lao population. Implementation of a simple carrier screening in pregnancy is practicable in the Lao PDR. PMID- 18414863 TI - Cytogenetic features in myelodysplastic syndromes. AB - Myelodysplastic syndromes (MDS) comprise a group of bone marrow diseases characterized by profound heterogeneity in morphologic presentation, clinical course, and cytogenetic features. Roughly 50% of patients display clonal chromosome abnormalities. In several multicentric studies, the karyotype turned out to be one of the most important prognostic parameters and was incorporated into statistical models aiming for a better prediction of the individual prognosis like the International Prognostic Scoring System. However, due to the profound cytogenetic heterogeneity, the impact of many rare abnormalities as well as combinations of anomalies occurring in a substantial portion of patients with MDS is still unknown and can only be delineated on the basis of large international multicentric cooperations. Recently, the German-Austrian MDS Study Group presented cytogenetic findings in 2,072 patients with MDS, which serve as a basis for the characterization of the cytogenetic subgroups discussed in this article. The availability of new therapeutic options for low- and high-risk MDS targeted against distinct entities characterized by specific chromosome abnormalities, like 5q-deletions, monosomy 7, and complex abnormalities underlines the important role of cytogenetics for the clinical management of MDS. This article thus focuses on the clinical and prognostic relevance, the molecular background, and therapeutic perspectives in these three cytogenetic subgroups. PMID- 18414864 TI - Phase 1 and pharmacokinetic study of weekly docosahexaenoic acid-paclitaxel, Taxoprexin, in resistant solid tumor malignancies. AB - PURPOSE: To determine the maximum tolerated dose, dose-limiting toxicity (DLT), and pharmacokinetics of weekly docosahexaenoic acid-paclitaxel (DHA-paclitaxel), a taxane fatty acid conjugate. EXPERIMENTAL DESIGN: Docosahexaenoic acid paclitaxel was administered by 2-hour i.v. infusion weekly for three out of four weeks. DHA-paclitaxel 200 mg/m(2) was dose escalated by 100 mg/m(2) per cohort to 600 mg/m(2). Blood samples for pharmacokinetics of DHA-paclitaxel and paclitaxel derived from DHA-paclitaxel were collected. RESULTS: Twenty-one patients received 42 cycles of treatment over five dose levels. Grade 3/4 neutropenia occurred in five patients but was not dose limiting. Grade 3 hyperbilirubinemia, a DLT, and grade 1 sensory neuropathy occurred at the highest dose level. PK analyses demonstrated dose proportional C (max) and AUC(0-24). Limited accumulation of DHA paclitaxel or paclitaxel occurred with weekly treatment. Increased DHA-paclitaxel and paclitaxel AUC(0-24) were associated with increased neutropenia. Of the 19 patients evaluable for response, three patients with esophageal, melanoma and colon carcinoma had stable disease for 11, 16, and 17 weeks, respectively. CONCLUSION: Docosahexaenoic acid-paclitaxel administered weekly to a maximum dose of 600 mg/m(2) was well-tolerated. The slow release of paclitaxel from DHA paclitaxel and the weekly schedule approximates continuous infusion paclitaxel which may be more active than every 3 week or weekly taxanes. PMID- 18414865 TI - Phase I clinical and pharmacokinetic study of oxaliplatin, irinotecan and capecitabine. AB - PURPOSE: To determine the maximum tolerated dose (MTD) and dose-limiting toxicity (DLT) of the combination of weekly oxaliplatin x 4, weekly irinotecan x 4 and capecitabine Monday through Friday for 4 weeks of every 6 week cycle in patients with solid tumors; to determine the pharmacokinetic profile of these agents in this combination; to observe patients for clinical anti-tumor response. METHODS: Twenty-two patients with metastatic solid tumors received oxaliplatin 60 mg/m(2) weekly x 4, irinotecan beginning at a dose of 40 mg/m(2) weekly x 4, and capecitabine Monday through Friday for 4 weeks of every 6 week cycle, initially at 1,000 mg twice daily (bid). RESULTS: The MTD was oxaliplatin 60 mg/m(2) weekly x 4, irinotecan 50 mg/m(2) weekly x 4 and capecitabine 450 mg bid Monday through Friday for 4 weeks of every 6 week cycle. One of six patients at this dose level developed DLT of nausea, vomiting, and diarrhea. Among patients treated with a constant capecitabine dose of 450 mg bid, there was a higher mean AUC of 5-FU in women than in men (mean +/- SD: 892 +/- 287 nM h vs. 537 +/- 182 nM h; Mann Whitney two-tailed, P = 0.02). There was one complete response in a patient with gastric cancer. CONCLUSION: The novel schedule of weekly oxaliplatin, weekly irinotecan, and capecitabine Monday through Friday, all administered for 4 weeks of every 6 week cycle, evaluated in this phase I trial is well-tolerated and demonstrated activity in a patient with gastric cancer. PMID- 18414866 TI - Modeling the joint epidemics of TB and HIV in a South African township. AB - We present a simple mathematical model with six compartments for the interaction between HIV and TB epidemics. Using data from a township near Cape Town, South Africa, where the prevalence of HIV is above 20% and where the TB notification rate is close to 2,000 per 100,000 per year, we estimate some of the model parameters and study how various control measures might change the course of these epidemics. Condom promotion, increased TB detection and TB preventive therapy have a clear positive effect. The impact of antiretroviral therapy on the incidence of HIV is unclear and depends on the extent to which it reduces sexual transmission. However, our analysis suggests that it will greatly reduce the TB notification rate. PMID- 18414867 TI - Characterization of the systems governing sexual and self-recognition in the white rot homobasidiomycete Amylostereum areolatum. AB - This study considered the systems controlling sexual and self-recognition in Amylostereum areolatum, a homobasidiomycetous symbiont of the Sirex woodwasp. To investigate the structure and organization of these systems in A. areolatum, we identified a portion of a putative homologue (RAB1) of the pheromone receptor genes of Schizophyllum commune and Coprinus cinereus, and a portion of a putative homologue of the S. commune mitochondrial intermediate peptidase (mip) gene. Diagnostic DNA-based assays for mating-type were developed and their application confirmed that the fungus has a heterothallic tetrapolar mating system. Segregation analysis showed that RAB1 is linked to mating-type B, while mip is linked to mating-type A. The results of sexual and vegetative compatibility tests suggest that sexual recognition in A. areolatum is controlled by two multiallelic mat loci, while self-recognition is controlled by at least two multiallelic het loci. Therefore, despite the association of A. areolatum with the woodwasp and the unique mixture of sexual and clonal reproduction of the fungus, both recognition systems of the fungus appear to be similar in structure and function to those of other homobasidiomycetes. This is the first report regarding the genes controlling recognition of a homobasidiomycete involved in an obligate mutualistic relationship with an insect. PMID- 18414868 TI - A mandibular osteolytic lesion in a systemic lupus erythematosus patient on long standing bisphosphonate therapy. PMID- 18414869 TI - Uni- and bidirectional wide angle CT colonography: effect on missed areas, surface visualization, viewing time and polyp conspicuity. AB - The effect of field of view on mucosal visualisation and reader efficiency during three-dimensional endoluminal CT colonography (CTC) was investigated. Twenty CTC datasets were reviewed at standard 90-degree and "wide" 140-degree viewing angles using customised viewing software (V3D colon; Viatronix), which listed number and size of missed mucosal areas ("missed regions tool") and percentage mucosal visualisation. We compared: (1) unidirectional and bidirectional flythrough using 140- versus 90-degree viewing angles; (2) reader analysis time comparing unidirectional 140-degree flythrough versus bidirectional 90-degree flythrough; (3) paired image snapshots of 12 polyps taken at each field of view were reviewed to assess conspicuity. All patients underwent conventional colonoscopy. Bidirectional 140-degree review reduced the numbers of missed areas by between eight- and 40-fold depending on size category, including those >1,000 mm(2), compared with standard 90-degree bidirectional flythrough (P < 0.001). Combined prone-supine unidirectional 140-degree flythrough and missed area review was 3.8 min faster than 90-degree bidirectional review (9.3 versus 5.5 min, P < 0.0001) for the same surface visualisation. When viewed as pairs, polyps were rated more conspicuous with a 90-degree field of view, P = 0.03. Wide-angle (140-degree) CTC can reduce both numbers of missed areas and review times. However, this may be at the expense of polyp conspicuity. PMID- 18414870 TI - Comparison of neutral oral contrast versus positive oral contrast medium in abdominal multidetector CT. AB - To determine whether neutral contrast agents with water-equivalent intraluminal attenuation can improve delineation of the bowel wall and increase overall image quality for a non-selected patient population, a neutral oral contrast agent (3% mannitol) was administered to 100 patients referred for abdominal multidetector row computed tomography (MDCT). Their results were compared with those of 100 patients given a positive oral contrast agent. Qualitative and quantitative measurements were done on different levels of the gastrointestinal tract by three experienced readers. Patients given the neutral oral contrast agent showed significant better qualitative results for bowel distension (P < 0.001), homogeneity of the luminal content (P < 0.001), delineation of the bowel-wall to the lumen (P < 0.001) and to the mesentery (P < 0.001) and artifacts (P < 0.001), leading to a significant better overall image quality (P < 0.001) than patients receiving positive oral contrast medium. The quantitative measurements revealed significant better distension (P < 0.001) and wall to lumen delineation (P < 0.001) for the patients receiving neutral oral contrast medium. The present results show that the neutral oral contrast agent (mannitol) produced better distension, better homogeneity and better delineation of the bowel wall leading to a higher overall image quality than the positive oral contrast medium in a non selected patient population. PMID- 18414871 TI - Measurement and determinants of infrarenal aortic thrombus volume. AB - Intra-luminal thrombus has been suggested to play a role in the progression of abdominal aortic aneurysm (AAA). The aims of this study were twofold. Firstly, to assess the reproducibility of a computer tomography (CT)-based technique for measurement of aortic thrombus volume. Secondly, to examine the determinants of infrarenal aortic thrombus volume in a cohort of patients with aortic dilatation. A consecutive series of 75 patients assessed by CT angiography with maximum aortic diameter > or = 25 mm were recruited. Intra-luminal thrombus volume was measured by a semi-automated workstation protocol based on a previously defined technique to quantitate aortic calcification. Intra- and inter-observer reproducibility were assessed using correlation coefficients, coefficient of variation and Bland-Altman plots. Infrarenal aortic thrombus volume percentage was related to clinical, anatomical and blood characteristics of the patients using univariate and multivariate tests. Infrarenal aortic thrombus volume was related to the severity of aortic dilatation assessed by total aortic volume (r = 0.87, P < 0.0001) or maximum aortic diameter (r = 0.74, P < 0.0001). We therefore examined the clinical determinates of aortic thrombus expressed as a percentage of total aortic volume. Aortic thrombus percentage was negatively correlated with serum high density lipoprotein (HDL, r = -0.31). By ordinal multiple logistic regression analysis serum HDL below median (< or = 1.2 mM: ) was associated with aortic thrombus percentage in the upper quartile adjusting for other risk factors (odds ratio 5.3, 95% CI 1.1-25.0). Infrarenal aortic thrombus volume can be measured reproducibly on CT. Serum HDL, which can be therapeutically raised, may play a role in discouraging aortic thrombus accumulation with implications in terms of delaying progression of AAA. PMID- 18414872 TI - Accuracy of ultrasound-guided, large-core needle breast biopsy. AB - Ultrasound-guided, large-core needle biopsy (US-LCNB) of suspicious breast lesions is acknowledged as less invasive and less expensive and less time consuming than surgical biopsy, and provides a histologic diagnosis with a comparable high degree. US-LCNB has been proven to help reduce the number of unnecessary surgeries for benign disease. Its limitations, however, are false negative results and underestimation of disease. Thus, the demand for breast teams is to carefully adhere to the principles of triple assessment and imaging histologic correlation, and follow-up of lesions with a specific benign histology after biopsy. Also, the acceptance of guidelines and rigorous quality controls help to reliably minimize the delay in the diagnosis of breast cancer in patients with false-negative biopsies. This paper aims to summarize the equipment and methods as well as the benefits and limitations of US-LCNB. Also, guidelines of quality assessment are suggested. Finally, recent developments which may help to overcome the limitations of US-LCNB will be discussed, i.e., directional vacuum assisted biopsy (VAB), three-dimensional (3D) US-guided biopsy, as well as the use of tissue harmonic imaging (THI) and compound imaging (CI) during biopsy. PMID- 18414873 TI - Reduction of dose to the female breast in thoracic CT: a comparison of standard protocol, bismuth-shielded, partial and tube-current-modulated CT examinations. AB - We evaluated the potential for reduction of dose to the female breast in computed tomography (CT) of the thorax by using three different techniques: bismuth shielding, partial CT scanning and tube-current modulation (TCM). Measurements and simulations of dose and image quality were performed for a 64-slice CT system using a semi-anthropomorphic thorax phantom with breasts added. Three-dimensional dose distributions were calculated by Monte Carlo (MC) methods. Noise was determined by measurements and simulations. Bismuth shielding resulted in a dose reduction of about 50% for the breast, noise increased up to 40% and image quality was impaired by artifacts. In partial CT scans, not irradiating the breasts directly, dose to the breasts was reduced typically by 50%. To sustain a constant noise level, an increase of irradiation in the anteroposterior position resulted in a higher dose to the spine. Reduction of dose to the breasts of about 10% was achieved with TCM; distribution of noise was homogeneous and image quality uniform. Reduction of dose to the female breast was achieved by using all adapted CT methods. Bismuth shielding may compromise image quality, increase noise level and introduce streak artifacts. Partial and TCM examinations reduced dose to the breast without influencing image quality. PMID- 18414874 TI - Color matching on natural substrates in cuttlefish, Sepia officinalis. AB - The camouflaging abilities of cuttlefish (Sepia officinalis) are remarkable and well known. It is commonly believed that cuttlefish-although color blind-actively match various colors of their immediate surroundings, yet no quantitative data support this notion. We assembled several natural substrates chosen to evoke the three basic types of camouflaged body patterns that cuttlefish express (uniform/stipple, mottle, and disruptive) and measured the spectral reflectance of the camouflaged pattern and the respective background using a fiber optic spectrometer. We demonstrate that the reflectance spectra of cuttlefish skin patterns correlate closely with the spectra of these natural substrates. Since pigmented chromatophores play a key role in cephalopod color change, we also measured the spectral reflectance of individual cuttlefish chromatophores under the microscope, and confirm the results from a previous publication reporting three distinct colors of chromatophores (yellow, orange, and dark brown) on the animals' dorsal side. Taken together, our results show that the color variations in substrate and animal skin can be very similar and that this may facilitate color match on natural substrates in the absence of color vision. PMID- 18414875 TI - Lactate dehydrogenase activity is increased in plasma of infants with advanced necrotizing enterocolitis. AB - In infants with necrotizing enterocolitis (NEC), intestinal gangrene defines advanced disease. Since intestinal ischemia is considered a pathogenetic factor for intestinal gangrene, serum activity of mucosal and seromuscular enzymes may be elevated in these patients. Our aim was to evaluate if serum enzymes activity is increased in infants with NEC associated with intestinal gangrene. We performed a retrospective review of the case notes of infants operated on for NEC between 1998 and 2006. Patients with preoperative determination of serum enzymes were included in the study, and were divided into Group A and Group B based on the presence or absence of intestinal gangrene, respectively. Serum activities of alkaline phosphatase (ALP), glutamic oxaloacetic transaminase (GOT), creatine kinase (CK), and lactate dehydrogenase (LDH) were compared in the two Groups. Values are medians (interquartile range). Thirty-five infants were operated on for NEC in the study period. Eighteen patients fulfilled the inclusion criteria: 12 in Group A and six in Group B. Group A patients had significantly higher LDH activity [1131.0 (1092.0-1300.0) vs. 482.0 (440.0-624.5) IU/L; P < 0.005]. Our findings suggest that LDH activity may be increased in infants with NEC and intestinal gangrene. Its evaluation could be a further tool in the surgical decision making process in infants with NEC. PMID- 18414876 TI - Physiologic predictors for the need for patch closure in neonatal congenital diaphragmatic hernia. AB - Technically expedient repair of CDH defects is desirable. With increasing trend toward thoracoscopic repair, herein we examine physiologic predictors for the need for patch closure (PC) versus primary closure. All neonates who underwent surgical repair of CDH defects in a geographically defined region between 1992 and 2002 were included (n = 210). Two groups of patients were compared, primary repair (PR) versus PC. The 25th quartile was used as a cut off point for continuous variables. Univariate and multivariate logistic regression were performed. One hundred and fifty neonates underwent open PR (71.43%) versus 28.57% had PC. On univariate analyses the following variables were significantly associated with the need for PC: prenatal diagnosis, birth weight <2.7 kg, gestational age <37 weeks, APGAR at 5 min <6, immediate postnatal PCO(2) >34, Immediate oxygen saturation <93%, use of Nitric oxide and the need for high frequency oscillation (HFO). On multivariate analyses, only a PCO(2) >34 and the need for HFO were significantly associated with PC. Neonates with an initial PCO(2) >34 or need HFO pre-operatively should be excluded from attempts to repair the CDH thoracoscopically based on their higher potential need for PC with its entailed technical difficulty and increased operative time. PMID- 18414877 TI - Papillary carcinoma of the thyroid gland in a child of thyrotoxicosis patient receiving radioactive iodine therapy: report of a case. AB - Herein we present the case of a 9-year-old girl who had an enlarged right lobe of the thyroid gland and sub-clinical hypothyroidism (thyroid stimulating hormone at 9.24 mIU/L). The patient had a history of unintentional exposure to radiation while her mother was receiving radionuclide therapy for diffuse toxic goiter. Ultrasonography of the young girl showed right lobe enlargement with diffuse coarse heterogenous echogenicity, compatible with a microcalcification pattern identified in both lobes of the thyroid gland. Histopathology of the tissue from a thyroidectomy revealed papillary thyroid carcinoma in the right lobe and chronic lymphocytic thyroiditis in the remaining tissue. Molecular pathology demonstrated an RET/PTC1 rearrangement in both tumor and non-tumorous tissue harboring thyroiditis. Considering the history of exposure and the characteristics of the thyroid pathology together, the PTC in this patient was likely a secondary-to-genetic alteration induced by external radiation. This case emphasizes the importance of stringent restrictions when giving radioactive iodine therapy to a patient with small children. PMID- 18414878 TI - Thoracotomies in children. AB - Thoracotomies in children have been less extensively studied, as the incidence of diseases necessitating thoracotomies is low in the pediatric age group. This study reviews childhood thoracic diseases, thoracotomy approaches, indications, and complications. Surgical procedures and complications of a total of 196 children below 16 years of age who underwent thoracotomy for various reasons at the Department of Thoracic Surgery, Ataturk Chest Diseases and Chest Surgery Training and Research Hospital, between January 2000 and December 2004, were reviewed in this study. Out of the 196 patients, 77 were female (39%) and 119 (61%) were male. The most commonly encountered indications for surgery were hydatid cyst (35%), bronchiectasis (25%), chronic nonspecific pleuritis (13%), chest wall deformities (10%), and mediastinal cystic formations and masses (10%). The other indications included tuberculosis (3%), aspergilloma (0.5%), fibrohyalinized cyst (0.5%), resection of trachea (0.5%), bronchogenic cyst (0.5%), inflammatory pseudo-tumor (0.5%), sequestration (1%), lipoblastoma (0.5%), and eosinophilic granuloma (1%). Out of the 196 patients, 176 underwent lateral thoracotomy and 20 patients with a chest wall deformity underwent midsternal incision. Complications were seen in 35 patients (18%): atelectasia and secretory retention (54%), wound infection (17%), hemorrhage (3%), chylothorax (3%), intrathoracic space (3%), and postoperative extended air leakage (20%). The mean hospital stay was 15 days and we did not encounter any mortality. The physiology and anatomy of the respiratory system and especially the respiratory control mechanism in pediatric patients vary from those of the adults, resulting in a more morbid course after thoracic surgery in children. Despite severe postoperative pain, posterolateral thoracotomy is the preferred approach in adults because of an advanced intrathoracic exposure and easy manipulation. On the other hand, lower pain threshold and the different types of diseases seen in children make lateral thoracotomy a more appropriate choice for thoracotomy, which, at the same time, spares the serratus anterior muscle decreasing its negative impact on postoperative respiratory function. PMID- 18414879 TI - Transluminal endoscopic electrosurgical incision of fenestrated duodenal membranes. AB - Duodenal fenestrated membranes are traditionally treated by side-to-side diamond shaped duodenoduodenostomy, or duodenotomy and resection. We describe an alternative endoscopic approach for its resolution. A flexible panendoscopy reaching the duodenal membrane was performed. A balloon was inserted to dilate its orifice. Traction was applied to the balloon to differentiate the border of the membrane forming the duodenal wall. After visualizing the ampulla, the membrane was incised using a sphincterotome or needle knife on two sites opposite to the bile duct. From May 2001 to August 2007, ten patients with a fenestrated duodenal membrane underwent transluminal endoscopic electrosurgical incision (TEEI). Mean patient age was 3.4 years (range 1 month to 15 years). The endoscopic procedure lasted from 30 to 60 min. Oral intake began 24 h postsurgery in eight patients and at 48 h postsurgery in two patients. Hospital stay lasted for 2-5 days. After 1 year of follow-up, eight patients were asymptomatic and thriving at present, and one had a double membrane, required a second endoscopy with TEEI, and has experienced occasional vomiting. An additional asymptomatic patient was lost after 3 months of follow-up. TEEI of fenestrated duodenal membranes is a feasible and effective procedure in children. PMID- 18414881 TI - Association of body mass index and clinical outcome 2 years after unicompartmental knee arthroplasty. AB - INTRODUCTION: Unicompartmental knee arthroplasty (UKA) has gained in popularity during the last years. However, the body mass index (BMI) of patients undergoing UKA is controversially discussed in the literature. There is, moreover, a paucity of information available concerning the association of BMI with early clinical outcomes after UKA. MATERIALS AND METHODS: We retrospectively reviewed the clinical data of 83 consecutive UKA, 2 years after surgery, and investigated the potential association of BMI and the outcome variables Knee Society Score (KSS), University of California at Los Angeles (UCLA) activity levels, anterior knee pain (AKP), range of motion, and implant failure. RESULTS: The KSS and UCLA significantly increased from 132 and 4.7 preoperatively to 187.5 and 7.1, respectively, after surgery. Knee flexion significantly improved from 123.7 to 128.4 degrees and the prevalence of extension deficiencies significantly decreased from 28.9 to 15.7%. Three knees (3.6%) failed and were converted to total knee arthroplasty. Failures were not associated with increased BMI (P = 0.387). The BMI had no significant association with KSS values, UCLA levels, and implant failure. We found a weak negative correlation between BMI and postoperative knee flexion (r = -0.285, P = 0.009) and a moderate positive correlation between BMI and the intensity of AKP (r = 0.525, P < 0.001). CONCLUSION: The results of the present study suggest that the BMI of patients undergoing UKA has no major impact on the early clinical outcome 2 years after surgery. There was, however, a definite correlation between the BMI and AKP. Longer follow-up is necessary to determine if overweight and obesity may increase revision rates after UKA. PMID- 18414880 TI - Characterization of antibodies that selectively detect alpha-synuclein in pathological inclusions. AB - Sensitive detection of alpha-synuclein (alpha-syn) pathology is important in the diagnosis of disorders like Parkinson's disease, dementia with Lewy bodies, and multiple system atrophy and in providing better insights into the etiology of these diseases. Several monoclonal antibodies that selectively react with aggregated alpha-syn in pathological inclusions and reveal extensive and underappreciated alpha-syn pathology in the brains of diseased patients were previously reported by Duda et al. (Ann Neurol 52:205-210, 2002). We sought to characterize the specificity of some of these antibodies (Syn 505, Syn 506 and Syn 514); using C-terminal and N-terminal truncations of alpha-syn, all three antibodies were determined to require N-terminal epitopes that minimally comprise amino acids 2-4, but possibly extend to amino acid 12 of alpha-syn. The selectivity of these antibodies was further assessed using biochemical analysis of human brains and reactivity to altered recombinant alpha-syn proteins with duplication variants of amino acids 1-12. In addition, by expressing wild-type or a double mutant (E46K/A53T) of alpha-syn in cultured cells and by comparing their immunoreactivities to another antibody (SNL-4), which has a similar primary epitope, it was determined that Syn 505, Syn 506 and Syn 514 recognize conformational variants of alpha-syn that is enhanced by the presence of the double mutations. These studies indicate that antibodies Syn 505, Syn 506 and Syn 514 preferentially recognize N-terminal epitopes in complex conformations, consistent with the dramatic conformational change associated with the polymerization of alpha-synuclein into amyloid fibrils that form pathological inclusions. PMID- 18414882 TI - Molecular identification of endocervical Chlamydia trachomatis infection among gestations at risk for preterm birth in Ecuador. AB - AIM AND METHOD: To identify Chlamydia trachomatis (CT) endocervical infection among gestations at risk for preterm birth by means of DNA amplification. RESULTS: One hundred fifty eight (n = 158) gestations of Guayaquil (63.3%) and Quito (36.7%) Ecuador, at risk for preterm birth: threatened preterm labor (TPL, 60.8%) and preterm premature rupture of membranes (PPROM, 39.2%) were recruited. CT infection was presented in 8.2% of cases (TPL: 10.4% and PPROM: 4.8%). No significant difference was observed in this frequency in regard to city or diagnosis. CONCLUSION: Although the causative role of CT infection for these antenatal complications cannot be completely ruled out, its presence may justify an antenatal routine screening program in order to provide appropriate therapy. PMID- 18414884 TI - Erroneous statement regarding 2D ultrasound fetal urinary bladder volume measurement. AB - The aim of the current paper is to inform the readers of an erroneous statement on the standard deviation (SD) when estimating the volume of fetal urinary bladder by 2D ultrasound. In the Archives of Gynecology and Obstetrics 2007; 276 (4), page 345-349, the authors Peixoto-Filho F.M. et al. refer to some studies presented by our group versus other studies regarding volume estimation of the fetal urinary bladder, in particular the ellipsoid method pioneered by Stuart Campbell in 1973 and the sum-of-cylinders method from our group. In our studies the volume estimation method was improved in three steps using methodological modifications. The SD was reduced from 17.3-10.9% to 12.9-5.5%, and 8.8-3.5% (bladder volumes 5-40 mL) The reductions were accompanied by P-values of 0.0270 (1 vs. 2) and 0.0543 (2 vs. 3), respectively. When comparing method 3 vs. 1 the P value was 0.0032. The SD reduction using method 3 vs. 2 did not reach statistical significance, however. Using this lack of significance on the 5% significance level, to prove that there were no improvements at all with our methodological modifications of the 2D ultrasound, as done by the authors, is misleading. PMID- 18414885 TI - Major labiaectomy as surgical management of vulvar lymphangioma circumscriptum: three cases and a review of the literature. AB - BACKGROUND: Vulvar lymphangioma circumscriptum (LC) is a developmental defect of lymphatics in deep dermal and subcutaneous layers. The entity may be acquired that induced by surgical intervention and radiation therapy for malignancy or congenital. Lymphangiomas of the vulva are rare. Thirteen cases of the congenital and 24 cases of acquired form have been reported in the literature presently. OBJECTIVE: To evaluate major labiaectomy as surgical management of vulval lymphangioma, we report our experiences on surgical treatment of three cases of this rare disease from Gynecologic Oncology Department of Vali-E-Asr Hospital. METHODS: Three female patients, aged 18, 17 and 63 years, presented with persistent edema, papules, and vesicles of the labia major, among which two first cases had congenital LC and another case had developed 9 years after radical hysterectomy, lymph node dissection, and adjuvant radiotherapy of cervical cancer. RESULT: Overall there was a clear improvement in the symptoms and signs associated with these conditions in all patients. In particular, all patients reported an improvement in the amount of edema following surgery. CONCLUSION: In particular, major labiaectomy seems to be more successful than methods such as lymphovenous anastomoses and lymph angioplasties. A single operation may prove beneficial; also, this approach has the potential to allow patients to rehabilitate to normal life and activity. PMID- 18414886 TI - A prospective trial of phaco-trabeculotomy combined with deep sclerectomy versus phaco-trabeculectomy. AB - BACKGROUND: Combined phacoemulsification, intraocular lens implantation, and trabeculectomy (PTE) is currently the standard procedure for most ophthalmic surgeons to treat uncontrolled open-angle glaucoma and cataract at one time. This study was designed to prospectively compare a new technique of phaco trabeculotomy plus deep sclerectomy (PDSTO) with standard phaco-trabeculectomy (PTE). METHODS: A consecutive series of 43 patients with uncontrolled open-angle glaucoma and cataract underwent combined glaucoma and cataract surgery. The procedure started as a two-site approach with phacoemulsification and IOL implantation through a temporal incision in clear cornea. Trabeculotomy and deep sclerectomy were performed in the superior quadrant. Trabeculectomy was also performed in the superior quadrant as a modified Cairns trabeculectomy. Postoperatively, examinations were performed on a daily base for 1 week. Follow up visits were applied 1, 3, 6, and 12 months after surgery. RESULTS: The mean preoperative intraocular pressure (IOP) was 26.5 mmHg (SD 7.8) for all patients enrolled. The mean IOP was 12.3 mmHg (SD 5.1) 1 day post surgery for the PTE group (p < 0.001) and 14.4 mmHg (SD 4.0) for the PDSTO group (p < 0.001). At 12 months post surgery the success rate according to the Advanced Glaucoma Intervention Study (AGIS), defined as an IOP lower than 18mmHg without medication, was 20% in the PTE group and 50% in the PDSTO group (p = 0.03). The number of postoperative complications was equally low for both groups. No severe complications, such as bleb infection, endophthalmitis, or choroidal hemorrhage were seen in this series. CONCLUSIONS: PDSTO offered significant IOP reduction and a success rate which was higher than that of the current standard, PTE. The specific intra- and postoperative complications of deep sclerectomy, trabeculotomy, and trabeculectomy were seen in our series, although the overall rate of postoperative complications proved low. PMID- 18414887 TI - Salivary duct carcinoma metastatic to eyelid and orbit-a case report. AB - BACKGROUND: Salivary duct carcinoma is a rare, extremely aggressive malignant tumor, demonstrating invasive growth with early regional and distant metastasis. We describe a case of parotid gland salivary duct carcinoma metastatic to the eyelid and orbit, as confirmed by immunohistochemical analysis. To the best of our knowledge, this is the first such case reported in the literature. METHODS: A 43-year-old male had left lower eyelid and left inferior fornix/anterior orbital masses. The patient underwent left lower eyelid and anterior orbital biopsies for histopathological evaluation. Immunohistochemical analysis for markers like androgen receptors, gross cystic disease fluid protein-15, cytokeratins, HER 2/neu, epithelial membrane antigen, S-100 proteins, progesterone receptors, and estrogen receptors were performed to establish diagnosis. RESULTS: Specimens from the eyelid and orbit revealed lobules of tumor cells exhibiting solid, micopapillary, and glandular appearance with central necrosis-comedo patterns. The tumor cells showed immunohistochemical reactivity to androgen receptor proteins, pankeratin, HER-2/neu and epithelial membrane antigen. Focal reactivity to gross cystic disease fluid protein-15 was also present. Immunoreactivity to S 100 proteins, progesterone receptors, and estrogen receptors were negative. Diagnosis was metastatic paratoid duct carcinoma to the left lower eyelid and left inferior orbit. CONCLUSIONS: Although metastatic SDC of the eyelid and orbit is a rare phenomenon, the possibility of this extremely aggressive entity should be included in the differential diagnosis when dealing with a patient with an eyelid and/or orbital tumor. PMID- 18414888 TI - Ocular toxicity of intravitreous adalimumab (Humira) in the rabbit. AB - PURPOSE: To evaluate the ocular toxicity of escalating doses of intravitreous adalimumab (Humira) in the rabbit eye. METHODS: Twelve New Zealand albino rabbits received unilateral intravitreous injections of 0.1 ml of adalimumab 0.25 mg (three eyes), 0.50 mg (three eyes), 1.0 mg (three eyes) or 0.1 ml balanced salt solution (BSS, three eyes). Slit-lamp biomicroscopy and fundoscopy were carried out at baseline, day 1, 7 and 14 following intravitreous injection, while electroretinography (ERG) was carried out at baseline and day 14. Animals were euthanized on day 14, and histopathological examination of the eyes was performed. RESULTS: Slit-lamp biomicroscopy and fundoscopy were normal in eyes having received BSS, 0.25 mg or 0.50 mg adalimumab; however, inflammation was present in two of three eyes having received 1.0 mg adalimumab. Similarly, comparison of scotopic and photopic ERG light at baseline and day 14 demonstrated no changes in eyes receiving BSS, 0.25 mg or 0.50 mg adalimumab, but two of three eyes having received 1.0 mg adalimumab showed a greater than 30% reduction in a and b wave. Finally, histopathology demonstrated no differences between eyes receiving BSS, 0.25 mg or 0.50 mg of adalimumab, but two of three eyes injected with 1.0 mg demonstrated inflammatory cell infiltration of the vitreous and anterior chamber, with one of these eyes demonstrating retinal necrosis. CONCLUSIONS: Escalating doses of intravitreous adalimumab in rabbit eyes caused no detectable functional or structural ocular toxicity up to a dose of 0.50 mg. Administration of 1.0 mg in 0.1 ml was associated with an inflammatory reaction and retinal necrosis. PMID- 18414889 TI - Are low inflammatory reactions involved in exudative age-related macular degeneration? Morphological and immunhistochemical analysis of AMD associated with basal deposits. AB - PURPOSE: Basal laminar and linear deposits (BLD) are associated with the development of choroidal neovascularization (CNV). Therefore, analysis of BLD composition may provide further information concerning the pathogenesis of BLD and CNV in age-related macular degeneration (AMD). METHODS: BLD in 25 specimens of surgically removed CNV were examined, using histochemical and immunohistochemical methods, for extracellular matrix proteins and their modulating enzymes, and for cell markers and proteins involved in inflammatory processes. In addition, ultrastructural electron microscopic analysis (EM) was performed. RESULTS: The chemical and structural composition of all the BLD was similar. Only the inner aspect of the BLD contained laminin and collagen IV, which corresponded to a new RPE basal lamina upon EM analysis. The extracellular matrix protein predominantly found in all layers of BLD was vitronectin, which was seen as a homogeneous material within the BLD upon EM analysis. The metalloproteinases MMP-2 and MMP-9 could only be detected in the inner aspect, while MMP-7 and TIMP-3 were observed predominantly in the outer aspect of BLD. In this area, staining for phospholipids and less intensely for neutral lipids was also visible. The labelling of complement complexes C3 and C5b-9 was intensely positive, and vascular endothelial growth factor (VEGF) was detected in all BLDs. CONCLUSIONS: Diffuse deposits such as BLD appear consistently with the development of CNV in AMD. They consist of extracellular matrix components and predominantly vitronectin. However, activated complement and VEGF could also be detected. The results of the current study may support the hypothesis that inflammatory processes are involved in the pathogenesis of BLD and CNV in AMD. PMID- 18414891 TI - Winching up heavy loads with a compliant arm: a new local joint controller. AB - A closed kinematic chain, like an arm that operates a crank, has a constrained movement space. A meaningful movement of the chain's endpoint is only possible along the free movement directions which are given implicitly by the contour of the object that confines the movement of the chain. Many technical solutions for such a movement task, in particular those used in robotics, use central controllers and force-torque sensors in the arm's wrist or a leg's ankle to construct a coordinate system (task frame formalism) at the local point of contact the axes of which coincide with the free and constrained movement directions. Motivated by examples from biology, we introduce a new control system that solves a constrained movement task. The control system is inspired by the control architecture that is found in stick insects like Carausius morosus. It consists of decentral joint controllers that work on elastic joints (compliant manipulator). The decentral controllers are based on local positive velocity feedback (LPVF). It has been shown earlier that LPVF enables contour following of a limb in a compliant motion task without a central controller. In this paper we extend LPVF in such a way that it is even able to follow a contour if a considerable counter force drags the limb away along the contour in a direction opposite to the desired. The controller extension is based on the measurement of the local mechanical power generated in the elastic joint and is called power controlled relaxation LPVF. The new control approach has the following advantages. First, it still uses local joint controllers without knowledge of the kinematics. Second, it does not need a force or torque measurement at the end of the limb. In this paper we test power-controlled relaxation LPVF on a crank turning task in which a weight has to be winched up by a two-joint compliant manipulator. PMID- 18414890 TI - The neuropeptide NAP provides neuroprotection against retinal ganglion cell damage after retinal ischemia and optic nerve crush. AB - BACKGROUND: NAP, an 8-amino acid peptide (NAPVSIPQ=Asn-Ala-Pro-Val-Ser-Ile-Pro Gln) derived from activity-dependent neuroprotective protein (ADNP), plays an important role in neuronal differentiation and the survival of neurons in different pathological situations. We already discovered that NAP increases the survival of retinal ganglion cells (RGC) in vitro, and supports neurite outgrowth in retinal explants at femtomolar concentrations. The aim of this study was to investigate the effects of NAP on RGC survival after transient retinal ischemia and optic nerve crush. METHODS: RGC of male Wistar rats were labelled retrogradely with 6 l FluoroGold injected stereotactically into both superior colliculi. Seven days later, retinal ischemia was induced by elevating the intraocular pressure to 120 mm Hg for 60 minutes or by crushing one optic nerve for 10 s after a partial orbitotomy. NAP was either injected intraperitoneally in the concentration of 100 microg/kg [corrected] 1 day before, directly after, and on the first and the second days after damage, or intravitreally (0.05 or 0.5 microg/eye) [corrected] directly after the optic nerve crush. Controls received the same concentrations of a control peptide. Densities of surviving RGC and activated microglial cells (AMC) were quantified in a masked fashion 10 days after damage by counting FluoroGold-labelled cells. RESULTS: After retinal ischemia, intraperitoneal injections of NAP increased the number of surviving RGC by 40% (p < 0.005) compared to the control group. After optic nerve crush, NAP raised the number of surviving RGC by 31% (p = 0.07) when injected intraperitoneally and by 54% (p < 0.05) when administered intravitreally. CONCLUSIONS: NAP acts neuroprotectively in vivo after retinal ischemia and optic nerve crush, and may have potential in treating optic nerve diseases. PMID- 18414892 TI - The scalable mammalian brain: emergent distributions of glia and neurons. AB - In this paper, we demonstrate that two characteristic properties of mammalian brains emerge when scaling-up modular, cortical structures. Firstly, the glia-to neuron ratio is not constant across brains of different sizes: large mammalian brains have more glia per neuron than smaller brains. Our analyses suggest that if one assumes that glia number is proportional to wiring, a particular quantitative relationship emerges between brain size and glia-to-neuron ratio that fits the empirical data. Secondly, many authors have reported that the number of neurons underlying one mm(2) of mammalian cortex is remarkably constant, across both areas and species. Here, we will show that such a constancy emerges when enlarging modular, cortical brain structures. Our analyses thus corroborate recent studies on the mammalian brain as a scalable architecture, providing a possible mechanism to explain some of the principles, constancies and rules that hold across brains of different size. PMID- 18414893 TI - Anasarca: not a nephrotic syndrome but dermatomyositis. AB - Juvenile dermatomyositis (JDM) is a rare autoimmune disease characterized by inflammation of the muscle, connective tissue, skin, gastrointestinal tract, and small nerves. Periorbital and facial edema may also be associated. Although localized edema is a common feature of JDM, generalized edema has rarely been reported. Here, we report a 3.5-year-old boy with JDM presenting with generalized edema. The diagnostic criteria of JDM rely on typical clinical manifestations that include: severe symmetric weakness of the proximal musculature, characteristic cutaneous changes, elevated serum skeletal muscle enzymes, and myopathic electromyographic pattern. Our patient initially received methylprednisolone and intravenous immunoglobulin (IVIG) without significant improvement, so he was given azathioprine and a prolonged course of oral prednisolone. We conclude that JDM should be suspected in patients presenting with anasarca in the absence of laboratory parameters of other causes of generalized edema and an appearance of heliotrope rash with muscle weakness. Also, we suggest that muscle magnetic resonance imaging (MRI) should be considered among the diagnostic tools of JDM. PMID- 18414894 TI - ACTH-dependent precocious pseudopuberty in an infant with DAX1 gene mutation. AB - DAX1 gene (Xp21) expression is involved in the development of the hypothalamo pituitary-gonadal and adrenal axes, and acts as a negative regulator of steroidogenesis. Mutations of this gene determine adrenal hypoplasia congenita (AHC) and hypogonadotropic hypogonadism. We report the case of a 9-month-old boy referred for the study of macrogenitosomia and pubic hair development. He had presented acute adrenal crises in the neonatal period and, later, a clinical picture of peripheral precocious puberty. A mutation in the DAX1 gene was found (Trp291Arg) and a diagnosis of AHC was made. Replacement doses of hydrocortisone (HC) (10 mg/m2/day) failed to produce a feedback inhibition of adrenocorticotropic hormone (ACTH), and testosterone levels remained high. Testosterone and ACTH values normalized after HC was progressively increased to 18 mg/m2/day. In conclusion, peripheral precocious puberty in patients with DAX1 gene mutations appears to be secondary to the stimulus exerted by ACTH on melanocortin receptors in Leydig cells and to the overexpression of testicular steroidogenesis activators by the loss of transcriptional repression. PMID- 18414895 TI - Prognostic significance of Survivin and CD44v6 in laryngeal cancer surgical margins. AB - OBJECTIVE: To evaluate the prognostic value of positive Survivin and CD44v6 expression in surgical margin after curative surgery for laryngeal cancer. METHODS: Immunohistochemistry was performed to detect the expression of Survivin and CD44v6 using polyclonal antibody on 112 pair of formalin-fixed, paraffin embedded laryngeal cancer and negative surgical margin tissue samples, and results were compared with data from the prospective registry of laryngeal cancer by univariate and multivariate logistic regression model focusing specifically on recurrence. The survival was assessed by the Kaplan-Meier method and proportional hazards model. RESULTS: Survivin and CD44v6 expression in surgical margins were documented in 39.3% (n = 44) and 31.3% (n = 35) patients and were associated with a higher incidence of tumor progression and poorer disease-free survival (P < 0.01). Multivariate logistic regression analysis demonstrated that Survivin (P = 0.000) and CD44v6 (P = 0.001) expression in surgical margin, T-stage (P = 0.006) and histological grade (P = 0.001) were independent predictive factors for recurrence. The proportional hazards model identified Survivin (P = 0.005) and CD44v6 (P = 0.004) expression in surgical margin, T-stage (P = 0.006) and histological grade (P = 0.001), as independently related to disease-free survival. Using subgroup analysis, we found that the Survivin+/CD44v6+ subgroup has the poorest prognosis, whereas the Survivin-/CD44v6- subgroup has the best prognosis (P = 0.000). CONCLUSIONS: Immunohistochemical assessment of both Survivin and CD44v6 status in negative surgical margin may be a valuable approach for predicting recurrence and survival after curative surgery for laryngeal cancer. PMID- 18414897 TI - Effect of temperature on development, eclosion, longevity and survivorship of malathion-resistant and malathion-susceptible strain of Culex quinquefasciatus. AB - Density of vectors and development of resistance against insecticides are two important aspects in the control of vector-borne diseases. Here, effect of temperature on different aspects of development of malathion-resistant and susceptible strains of Culex quinquefasciatus was evaluated in the laboratory. Fourth-instar larvae of C. quinquefasciatus were exposed to 37 degrees C, 39 degrees C, 40 degrees C, and 41 degrees C, and their LT(50) values calculated. The fourth-instar larvae were subjected to heat for 4 h at 39 degrees C and live larvae were reared until the completion of life cycle. The larvae of malathion resistant strain were more tolerant to heat than the larvae of malathion susceptible ones. The difference in mortality between 37 degrees C, 39 degrees C, 40 degrees C, and 41 degrees C of both the strains of C. quinquefasciatus were highly significant. Pupation percentage of heat-exposed larvae of both the strains was higher and delayed than the control ones. The percentage of adult emergence from the heat-exposed larvae was significantly lower than the control. The adult female of malathion-resistant strain showed increased longevity when exposed to heat at larval stage but reverse was true in case of malathion susceptible. The highlights of the study would help in the management of resistance of mosquito vectors in the tropics where variable climatic condition is observed. PMID- 18414896 TI - The role of cell differentiation in controlling cell multiplication and cancer. AB - It has been suggested that cancer ought to be regarded as a disease of cell differentiation. In multicellular organisms, indeed, the control of cell multiplication is linked to cell specialization: During the process of differentiation embryonic cells, while cycling, acquire the ability to perform specialized functions. This ability is incompatible with cell cycling which, as a consequence, is repressed with forthcoming differentiation. Thus, the number of amplification divisions that occur during the period while differentiation is proceeding decides on the number of specialized cells produced. The progress in differentiation, contrary to usual assumptions, is accompanied by an increase in the cellular content of cytoplasm. The reason must be that cell specialization requires a specific amount and array of membrane-bounded cytoplasmic structures. Since the multiplication of these structures depends on membranous templates, their amount increases only if more cytoplasm is produced per cycle than required for a doubling, thus constituting an intracellular timer of differentiation: The larger the net rate of cytoplasmic growth per cell cycle, the fewer cycles occur. Extracellular signals modulate cell multiplication by altering the net rate of cytoplasmic growth. Each persisting alteration, however, that reduces this rate to zero, prevents differentiation, and thus causes the cells to retain embryonic capabilities and to initiate cancer. Cancer cells can be induced to differentiate and cease proliferation by support of cytoplasmic growth. This corroborates the suggestion that cancer must be regarded as a disease of cell differentiation and our conclusion that cancer is caused by a deficiency in cytoplasmic growth. Support of the latter must be an efficient principle in cancer therapy although limited by the organism's dependence on cell renewal. PMID- 18414898 TI - IFNG +874T/A, IL10 -1082G/A and TNF -308G/A polymorphisms in association with tuberculosis susceptibility: a meta-analysis study. AB - Susceptibility to infectious diseases is influenced by genetic background and efficient cellular immune activation is responsible for protection. In tuberculosis (TB), interferon-gamma (IFNgamma) is crucial to control intracellular growth of Mycobacterium tuberculosis while interleukin-10 (IL-10) has an antagonistic role. Tumor necrosis factor (TNF) is a central mediator of granuloma formation and control of bacilli spread synergizing with IFNgamma to hamper M. tuberculosis infection. Single nucleotide polymorphisms (SNPs) located at these genes could influence cytokine levels and regulate resistance and susceptibility to TB. The aim of this study was to determine the association of the interferon-gamma gene (IFNG) +874T/A, interleukin-10 gene (IL10) -1082G/A and tumor necrosis factor gene (TNF) -308G/A SNPs with TB in several populations using meta-analysis. We searched for association studies correlating these polymorphisms and TB using pre-established keywords in Medline. Meta-analysis was conducted with random effects models to account for heterogeneity between studies. Eleven studies were included in the IFNG +874T/A meta-analysis, while eight were used for the IL10 -1082G/A, and 10 were employed for TNF -308G/A. Data were analyzed in respect to associations between alleles, genotypes and minor allele carriers. Statistically significant results were found only for IFNG. The +874T allele of IFNG showed a protective significant association (OR = 0.75; 95% CI, 0.634-0.887; P = 0.0008). Though not significant, IL10 presented a trend towards protection when only studies with pulmonary TB patients were considered. This data reinforces the critical importance of IFNG +874T/A as a genetic marker for TB resistance and this information can be used for better design of a TB vaccine. PMID- 18414899 TI - Foliar delta(15)N values characterize soil N cycling and reflect nitrate or ammonium preference of plants along a temperate grassland gradient. AB - The natural abundance of stable (15)N isotopes in soils and plants is potentially a simple tool to assess ecosystem N dynamics. Several open questions remain, however, in particular regarding the mechanisms driving the variability of foliar delta(15)N values of non-N(2) fixing plants within and across ecosystems. The goal of the work presented here was therefore to: (1) characterize the relationship between soil net mineralization and variability of foliar Deltadelta(15)N (delta(15)Nleaf - delta(15)Nsoil) values from 20 different plant species within and across 18 grassland sites; (2) to determine in situ if a plant's preference for NO (3) (-) or NH (4) (+) uptake explains variability in foliar Deltadelta(15)N among different plant species within an ecosystem; and (3) test if variability in foliar Deltadelta(15)N among species or functional group is consistent across 18 grassland sites. Deltadelta(15)N values of the 20 different plant species were positively related to soil net mineralization rates across the 18 sites. We found that within a site, foliar Deltadelta(15)N values increased with the species' NO (3) (-) to NH (4) (+) uptake ratios. Interestingly, the slope of this relationship differed in direction from previously published studies. Finally, the variability in foliar Deltadelta(15)N values among species was not consistent across 18 grassland sites but was significantly influenced by N mineralization rates and the abundance of a particular species in a site. Our findings improve the mechanistic understanding of the commonly observed variability in foliar Deltadelta(15)N among different plant species. In particular we were able to show that within a site, foliar delta(15)N values nicely reflect a plant's N source but that the direction of the relationship between NO (3) (-) to NH (4) (+) uptake and foliar Deltadelta(15)N values is not universal. Using a large set of data, our study highlights that foliar Deltadelta(15)N values are valuable tools to assess plant N uptake patterns and to characterize the soil N cycle across different ecosystems. PMID- 18414900 TI - Maintenance therapy with mycophenolate mofetil for children with severe lupus nephritis after low-dose intravenous cyclophosphamide regimen. AB - Although recent studies on adults with lupus nephritis indicate that mycophenolate mofetil (MMF) may be effective in maintaining remission for patients who previously received short-term intravenous cyclophosphamide (IVCY) induction therapy, the experience with the new immunosuppressive agent in children with severe lupus nephritis has not been as satisfactory thus far. To assess the efficacy and safety of maintenance therapy with MMF, we prospectively analyzed four patients with biopsy-proven severe lupus nephritis (three girls, one boy; mean age 12 years; two with class IIIA, two with class IVG(A); mean duration of lupus nephritis 7 months) receiving MMF for at least 6 months after induction treatment. These patients had been treated previously with 6 months of low-dose IVCY combined with oral mizoribine and steroids for induction, followed by therapy with MMF adjusted to maintain predose mycophenolic acid (C0-MPA) levels at 2-5 mcg/ml. Mean follow-up after staring MMF was 27.5 months (range 6 41). The mean MMF dose required was 405 +/- 49 mg/m(2) per 12 h, which maintained mean C0-MPA levels of 3.3 +/- 0.41 mcg/ml. No patient experienced renal flares during maintenance therapy with MMF, which permitted a significant reduction in mean prednisolone dose from 11.9 +/- 1.3 to 3.9 +/- 2.6 mg/day (P = 0.003). No significant gastrointestinal or hematologic side effects of MMF were noted. This preliminary study demonstrates that maintenance therapy with MMF after a low-dose IVCY regimen appears to be a promising intervention without adverse effects in children with severe lupus nephritis. These data should be confirmed by a prospective randomized multicenter clinical trial. PMID- 18414902 TI - Fear of Cancer Recurrence Inventory: development and initial validation of a multidimensional measure of fear of cancer recurrence. AB - BACKGROUND: Despite the fact that the fear of cancer recurrence is to varying degrees almost universal in cancer survivors, there is a lack of validated multidimensional instruments to evaluate this issue specifically. PURPOSE: The goal of this study was to develop and empirically validate a multidimensional self-report scale for assessing the fear of cancer recurrence, the Fear of Cancer Recurrence Inventory (FCRI). METHODS: A provincial medical databank was used to randomly select a pool of 1,704 French-Canadian patients who had been treated for breast, prostate, lung, and colorectal cancer within the past 10 years. Of these, 300 patients were asked to complete the FCRI on two occasions. RESULTS: The factorial analysis conducted on the final 42-item scale revealed a seven component solution (64% of the variance) including the following factors: triggers, severity, psychological distress, coping strategies, functioning impairments, insight, and reassurance. The results also supported the internal consistency (alpha = 0.95) and the temporal stability (r = 0.89) of the FCRI, as well as its construct validity with other self-report scales assessing fear of cancer recurrence (r = 0.68 to 0.77) or related constructs such as psychological distress (r = 0.43 to 0.77) and quality of life (r = -0.20 to -0.36). CONCLUSIONS: This study suggests that the French-Canadian version of the FCRI is a reliable and valid instrument for evaluating the multidimensional aspects of the fear of cancer recurrence. PMID- 18414901 TI - Chronic kidney disease after liver, cardiac, lung, heart-lung, and hematopoietic stem cell transplant. AB - Patient survival after cardiac, liver, and hematopoietic stem cell transplant (HSCT) is improving; however, this survival is limited by substantial pretransplant and treatment-related toxicities. A major cause of morbidity and mortality after transplant is chronic kidney disease (CKD). Although the majority of CKD after transplant is attributed to the use of calcineurin inhibitors, various other conditions such as thrombotic microangiopathy, nephrotic syndrome, and focal segmental glomerulosclerosis have been described. Though the immunosuppression used for each of the transplant types, cardiac, liver and HSCT is similar, the risk factors for developing CKD and the CKD severity described in patients after transplant vary. As the indications for transplant and the long term survival improves for these children, so will the burden of CKD. Nephrologists should be involved early in the pretransplant workup of these patients. Transplant physicians and nephrologists will need to work together to identify those patients at risk of developing CKD early to prevent its development and progression to end-stage renal disease. PMID- 18414903 TI - Expression of CD11b/CD18 adhesion molecules on circulating phagocytes-a novel aid to diagnose infection in patients with cancer. AB - GOALS OF WORK: No blood marker available to date is useful for distinguishing infection-related from neoplasm-related fever. We evaluated the expression of the peripheral blood phagocyte CD11b/CD18 adhesion molecule complex for this purpose. MATERIALS AND METHODS: Neutrophil and monocyte CD11b/CD18 expression was assessed in two cohorts of patients with advanced solid cancer (n = 120) and in healthy controls (n = 63). The cancer series included 89 patients with verified infection, 23 without infection, and eight with neoplastic fever. CD11b/CD18 expression was measured using flow cytometry, and serum C-reactive protein (CRP) concentration was determined with immunoturbidimetric assay. RESULTS: Cancer patients with infection had higher blood neutrophil and monocyte CD11b/CD18 expression levels than patients with neoplastic fever, those with advanced cancer without infection, or healthy controls (p < 0.01 for all analyses). High CD11b/CD18 values were measured exclusively in individuals diagnosed with infection. Receiver-operating characteristic area under the curve (AUC) for neutrophil and monocyte CD11b/CD18 expression for the discrimination of infection from neoplastic fever was 0.80 (95% CI, 0.70 to 0.88), which was superior (p = 0.039 and p = 0.049, respectively) to serum CRP on admission (AUC 0.51, 0.40 to 0.62). CONCLUSIONS: Peripheral blood phagocytic cell CD11b/CD18 expression is useful for making a differential diagnosis between infection and neoplasm-related fever in cancer patients. PMID- 18414904 TI - Functional evaluation of conventional 'Celsite' venous ports versus 'Vortex' ports with a tangential outlet: a prospective randomised pilot study. AB - GOAL OF WORK: Totally implantable venous access ports are widely accepted in cancer patient treatment, but withdrawal occlusion (WO) can hamper the use of the device. A newly designed Vortex VX port, with a tangential outlet, should allow better clearance of the chamber, thereby reducing occlusion of the device. The present study compared the Vortex port to the classically shaped Celsite port with regards to functional complications. MATERIALS AND METHODS: Two hundred cancer patients were included in a prospective, randomised controlled trial and randomly assigned to the implantation of a Vortex or a Celsite port. Insertion details such as used vein, catheter tip position and infusion or aspiration abilities were recorded. Data were collected concerning ease of access, and functional evaluation was performed each time the port was accessed, regarding the ability to infuse fluids with a syringe and to withdraw blood by measuring the filling time of a Vacutainer blood tube. MAIN RESULTS: Ninety-nine patients received a Celsite port, and 101 had a Vortex port. Demographic variables and insertion details were comparable in both groups. All functional complications, including WO, total occlusion, sluggish inflow and sluggish withdrawal, were higher in the Celsite group (16.12%) than in the Vortex group (11.36%). This difference was not statistically significant. CONCLUSIONS: This study revealed that functional problems occurred less frequently in Vortex compared to Celsite ports. Differences were small and not significant, which indicates that functional problems may be related to other factors. PMID- 18414906 TI - New tuber species found in Poland. AB - New information from a survey of truffles carried out in southern part of Poland in September 2007 is presented. The fruit bodies of Tuber aestivum, T. excavatum, T. rufum and one unidentified Tuber sp. were found. The soil chemistry of the five T. aestivum sites was analysed. Our inventory showed that T. aestivum prefers the mixed forest with host-trees such as: Quercus robur, Corylus avellana, Carpinus betulus, Fagus silvaticus and Tilia cordata. Selection of local inocula of T. aestivum for the propagation of truffles production could be an alternative to the Mediterranean truffles orchards in times when demand for the fungi will be increasing. Fruit bodies of truffles are conserved in the collection of biological material in the Forest Research Institute in Sekocin Stary. PMID- 18414905 TI - Maintenance of effects of a home-based physical activity program among breast cancer survivors. AB - PURPOSE: Although physical activity (PA) adoption improves fitness and psychological well-being among cancer survivors, PA maintenance has not been examined. This paper presents follow-up of a home-based PA program for women treated for early-stage breast cancer. MATERIALS AND METHODS: Eighty-six sedentary women (mean age = 53.14 years, SD = 9.70) were randomly assigned to a PA or contact control group. The PA group received a 12-week telephone counseling program to adopt PA. Assessments were conducted at baseline, end-of-intervention (12 weeks), 6, and 9 months post-baseline. RESULTS: When comparing change from end-of-intervention (12 weeks) between groups, a significant reduction was observed in minutes of PA at 6 months (t = -2.10, p < 0.05), but there was no decrease in intervention effect at 9 months (t = -0.19, p = 0.84). Similarly, post-intervention reductions in fatigue were lost at 6 months (t = 3.27, p < 0.01), but remained present at 9 months (t = 1.65, p = 0.10). PA group's fitness improvements were maintained at both follow-ups (t = 1.04, p = 0.30 and t = 0.05, p = 0.96). The previously significant intervention effect on vigor was maintained at 6 months (t = 1.32, p = 0.19) but was significantly reduced at 9 months (t = 2.15, p < 0.05). PA participants were more likely to progress in motivational readiness at 6 (OR = 5.95, 95% CI = 2.30, 15.36) and 9 months (OR = 4.09, 95% CI = 1.69, 9.87); however, group differences in meeting PA guidelines were not maintained. CONCLUSION: Some positive effects of a home-based PA intervention for breast cancer patients were maintained at 6 and 9 months. PMID- 18414907 TI - Geometric and electronic structures of new endohedral fullerenes: Eu@C72. AB - The geometric and electronic structures of rare earth metallofullerenes Eu@C72 were investigated using density functional theory (DFT) within a generalized gradient approximation (GGA). The geometric optimization revealed that the most favorable endohedral site for Eu is off-center along the C2 axis on the sigma(v) plane pointing to the (5, 5) bond at the fusion of two pentagons. Calculations for electronic structures show that two 6s electrons in Eu transfer to the lowest unoccupied-molecular orbitals of C72 while 4f electrons remain in Eu. PMID- 18414908 TI - Probing the interactions of the solvated electron with DNA by molecular dynamics simulations: bromodeoxyuridine substituted DNA. AB - Solvated electrons ((e-)(aq)) are produced during water radiolysis and can interact with biological substrates, including DNA. To augment DNA damage, radiosensitizers such as bromo-deoxyuridine (BUdR), often referred to as an "electron affinic radiosensitizer", are incorporated in place of isosteric thymidine. However, little is known about the primary interactions of (e-)(aq) with DNA. In the present study we addressed this problem by applying molecular modeling and molecular dynamics (MD) simulations to a system of normal (BUdR.A) DNA and a hydrated electron, where the excess electron was modeled as a localized (e-)(H2O)6 anionic cluster. Our goals were to evaluate the suitability of the MD simulations for this application; to characterize the motion of (e-)(aq) around DNA (e.g., diffusion coefficients); to identify and describe configurational states of close (e-)(aq) localization to DNA; and to evaluate the structural dynamics of DNA in the presence of (e-)(aq). The results indicate that (e-)(aq) has distinct space-preferences for forming close contacts with DNA and is more likely to interact directly with nucleotides other than BUdR. Several classes of DNA - (e-)(aq) contact sites, all within the major groove, were distinguished depending on the structure of the intermediate water layer H-bonding pattern (or its absence, i.e., a direct H-bonding of (e-)(aq) with DNA bases). Large-scale structural perturbations were identified during and after the (e-)(aq) approached the DNA from the major groove side, coupled with deeper penetration of sodium counterions in the minor groove. PMID- 18414909 TI - Dysregulation of large-conductance Ca2+-activated K+ channel expression in nonsyndromal mental retardation due to a cereblon p.R419X mutation. AB - A nonsense mutation (R419X) in the human cereblon gene [mutation (mut) CRBN] causes a mild type of autosomal recessive nonsyndromal mental retardation (ARNSMR). CRBN, a cytosolic protein, regulates the assembly and neuronal surface expression of large-conductance Ca(2+)-activated K(+) channels (BK(Ca)) in brain regions involved in memory and learning. Using the real-time quantitative polymerase chain reaction, we show that mut CRBN disturbs the development of adult brain BK(Ca) isoforms. These changes are predicted to result in BK(Ca) channels with a higher intracellular Ca(2+) sensitivity, faster activation, and slower deactivation kinetics. Such alterations may contribute to cognitive impairments in patients with mild ARNSMR. PMID- 18414910 TI - Biliary dilatation and duodenal intussusception secondary to enteric duplication cyst: MDCT diagnosis. AB - Enteric duplication cysts are rare congenital anomalies that may occur anywhere along the gastrointestinal tract, most commonly involving the small bowel. The distal ileum, jejunum, and duodenum are affected in descending order of frequency. We describe a case of biliary dilatation and duodenal intussusception caused by an enteric duplication cyst in an adult patient. To our knowledge, there are no other reported cases of this entity in an adult in the English literature. Multidetector computed tomography (MDCT) findings are emphasized, and the value of multiplanar reformation (MPR) in forming a correct preoperative differential diagnosis is discussed. PMID- 18414912 TI - Diversification of Lrk/Tak kinase gene clusters is associated with subfunctionalization and cultivar-specific transcript accumulation in barley. AB - Lrk (Lr10 receptor-like kinase) and Tak (Triticum aestivum kinase) belong to the receptor-like kinase (RLK) supergene family in higher plants. Three Lrk/Tak gene regions spanning greater than 600 kb were identified via a genome-wide survey of barley gene-rich BAC clones. Two Lrk/Tak gene clusters are positioned on barley chromosome 3 (3H) and another is localized on chromosome 5 (1H), with each Lrk and Tak open reading frame physically positioned in a back-to-back orientation. Thirteen new Lrk/Tak-like fragments were cloned from the two clusters on 3H and the single cluster on 1H, respectively, and compared phylogenetically with other grass Lrk/Tak-like genes, including a 280-kb Lrk/Tak cluster on rice chromosome 1S. Physically clustered Lrk/Tak-like genes always form monophyletic groups; this suggests that the primary mechanism of expansion of the Lrk/Tak RLK super family was by tandem duplication, of which most members were duplicated after speciation of the Poaceae. Cultivar-dependent transcript accumulation of some Lrk/Tak family members on 3H, as revealed via Barley1 GeneChip microarray analysis, is consistent with the hypothesis of subfunctionalization of Lrk/Tak members following tandem duplication. PMID- 18414913 TI - Using a short-term parameter of heart rate variability to distinguish awake from isoflurane anesthetic states. AB - The measurement of anesthetic depth is important in anesthesiology. Although heart rate variability (HRV) is profoundly affected by general anesthesia, it has not yet been commonly used in this field. One of the reasons is the lack of suitable parameters of HRV for short-term observations. In this study, we designed a time domain parameter of HRV named the similarity index. It was based on observing the trend of the distribution of instantaneous heart rates as time moved. Taking epochs of ECG data as short as 64 s can derive the index. We observed the values of this index of 30 patients when they were awake and under isoflurane anesthesia. The values had very little overlapping between the two states and the prediction probability to distinguish the two states was 0.91. We suggest that HRV, if suitably treated, can play more roles in the monitoring of anesthetic depth. PMID- 18414911 TI - Expression patterns in soybean resistant to Phakopsora pachyrhizi reveal the importance of peroxidases and lipoxygenases. AB - Soybean rust caused by Phakopsora pachyrhizi Sydow is a devastating foliar disease that has spread to most soybean growing regions throughout the world, including the USA. Four independent rust resistance genes, Rpp1-Rpp4, have been identified in soybean that recognize specific isolates of P. pachyrhizi. A suppressive subtraction hybridization (SSH) complementary DNA (cDNA) library was constructed from the soybean accession PI200492, which contains Rpp1, after inoculation with two different isolates of P. pachyrhizi that result in susceptible or immune reactions. Both forward and reverse SSH were performed using cDNA from messenger RNA pooled from 1, 6, 12, 24, and 48 h post inoculation. A total of 1,728 SSH clones were sequenced and compared to sequences in GenBank for similarity. Microarray analyses were conducted on a custom 7883 soybean-cDNA clone array encompassing all of the soybean-rust SSH clones and expressed sequence tags from four other soybean cDNA libraries. Results of the microarray revealed 558 cDNA clones differentially expressed in the immune reaction. The majority of the upregulated cDNA clones fell into the functional category of defense. In particular, cDNA clones with similarity to peroxidases and lipoxygenases were prevalent. Downregulated cDNA clones included those with similarity to cell-wall-associated protein, such as extensins, proline-rich proteins, and xyloglucan endotransglycosylases. PMID- 18414914 TI - MBEC special issue on microcirculation "engineering principles of vascular networks". PMID- 18414915 TI - Evolution of a multi-agent system in a cyclical environment. AB - The synchronisation phenomena in biological systems is a current and recurring subject of scientific study. This topic, namely that of circadian clocks, served as inspiration to develop an agent-based simulation that serves the main purpose of being a proof-of-concept of the model used in the BitBang framework, that implements a modern autonomous agent model. Despite having been extensively studied, circadian clocks still have much to be investigated. Rather than wanting to learn more about the internals of this biological process, we look to study the emergence of this kind of adaptation to a daily cycle. To that end we implemented a world with a day/night cycle, and analyse the ways the agents adapt to that cycle. The results show the evolution of the agents' ability to gather food. If we look at the total number of agents over the course of an experiment, we can pinpoint the time when reproductive technology emerges. We also show that the agents adapt to the daily cycle. This circadian rhythm can be shown by analysing the variation on the agents metabolic rate, which is affected by the variation of their movement patterns. In the experiments conducted we can observe that the metabolic rate of the agents varies according to the daily cycle. PMID- 18414917 TI - [Postoperative delirium in the critically ill]. AB - Acute confusional states (delirium) occur in up to 80% of patients in the intensive care unit. Delirium is an important independent prognostic determinant of hospital outcome, including duration of mechanical ventilation, nursing home placement, functional decline and death. Additionally, it may herald systemic diseases such as sepsis and multi-organ failure. Recently, a number of new screening instruments have been validated for the monitoring of delirium in non communicative patients receiving mechanical ventilation. Critical care patients should be routinely assessed for delirium and treated immediately using available preventive and therapeutic measures, both pharmacological and non pharmacological, to improve the clinical course. PMID- 18414916 TI - [Intrathecal morphine in orthopaedic surgery patients. Optimised dose in patients receiving dipyrone]. AB - INTRODUCTION: The influence of different postoperative doses of intrathecal morphine on the time of first opioid request by orthopaedic patients was investigated. The first choice analgesic was dipyrone and a maximum dose of 6 mg/day was allowed. METHODS: A prospective, double-blind, placebo-controlled, clinical trial was conducted with 15 patients in each group receiving intrathecally either a placebo, 0.05 mg morphine, 0.1 mg morphine or 0.2 mg morphine in combination with a spinal anaesthesia with isobaric bupivacaine. RESULTS: The number of patients without opioid requirement during the first 24 h after surgery were 3, 8, 14 and 14 in the placebo group and after 0.05 mg (p=0.128), 0.1 mg (p=0.0001) and 0.2 mg (p=0.0001) intrathecal morphine, respectively. The average time until first opioid requirement increased in a dose dependent manner from 10.3 h to 23.9 h (p<0.0001). CONCLUSION: In orthopaedic patients with dipyrone as the primary analgesic, the addition of 0.1 mg or 0.2 mg morphine to spinal anaesthesia provided a simple long-lasting postoperative analgesia and the use of additional opioids could be avoided during the 24h postoperative period. PMID- 18414919 TI - Spatial perception and knowledge. PMID- 18414918 TI - Investigations on the solubility of corrosion products on depleted uranium projectiles by simulated body fluids and the consequences on dose assessment. AB - Ingestion and inhalation of corrosion products covering weathered penetrators made of depleted uranium (DU) represent potential radiological exposure pathways. In order to study the bioavailability of these corrosion products, their solubility was determined using simulated gastric and pulmonary juices. About 75 and 36% of the uranium in the corrosion products were found to be soluble in simulated gastric and pulmonary juices, respectively. The effective dose coefficient for adults after ingestion was calculated to be 0.61 muSv mg(-1) DU. This compares to an effective dose coefficient for an adult of 0.71 muSv mg(-1) for DU materials given by the World Health Organization (WHO). The effective dose coefficient for inhalation was calculated to be 3.7 x 10(-6 )Sv Bq(-1) for workers and 5.3 x 10(-6 )Sv Bq(-1) for members of the public, respectively, which is between those of particles of Types M and S as defined by the International Commission on Radiological Protection (ICRP). The speciation of the corrosion products was investigated by time-of-flight secondary ion mass spectrometry (TOF SIMS). The mean oxidation state of uranium was found to be 4.6, which suggests that the uranium in the corrosion products consists of a mixture of U(IV) and U(VI) species. PMID- 18414920 TI - Fracture risk associated with different types of oral corticosteroids and effect of termination of corticosteroids on the risk of fractures. AB - We conducted a case-control study on fracture risk associated with the use of orally administered prednisolone/prednisone, budesonide, methylprednisolone, and hydrocortisone to assess if the various preparations were associated with different fracture patterns. Cases were all subjects with any fracture sustained during the year 2000 (n = 124,655). For each case, three controls (n = 373,962) matched on age and gender were randomly drawn from the background population. Adjustments were made for concurrent diseases, use of other drugs, and a number of other factors. Oral prednisolone/prednisone was associated with a dose dependent increase in fracture risk starting from a dose of around 6.7 mg/day. Oral budesonide was not associated with an increase in overall fracture risk, but the doses in general were low (<3 mg/day). Oral hydrocortisone was not associated with overall risk of fractures. Oral methylprednisolone was only used intermittently and was not associated with an increase in overall fracture risk at the low doses used. After termination of oral prednisolone/prednisone, it took more than 1 year for fracture risk to return to the levels of the background population. Oral prednisolone is associated with a dose-dependent increase in overall fracture risk. Budesonide at low doses did not seem to be associated with fracture risk. Hydrocortisone was not associated with an increase in the risk of fractures. It may take a year from last use of prednisolone/prednisone before fracture risk returns to that of the general population. PMID- 18414922 TI - Electrophysiological and fluorescence microscopy studies with HERG channel/EGFP fusion proteins. AB - HERG (human ether-a-go-go-related gene) encodes the Kv11.1 protein alpha-subunit that underlies the rapidly activating delayed rectifier K+ current (IKr) in the heart. Alterations in the functional properties or membrane incorporation of HERG channels, either by genetic mutations or by administration of drugs, play major roles in the development of life-threatening torsades de pointes cardiac arrhythmias. Visualization of ion channel localization is facilitated by enhanced green fluorescent protein (EGFP) tagging, but this process can alter their properties. The aim of the present study was to characterize the electrophysiological properties and the cellular localization of HERG channels in which EGFP was tagged either to the C terminus (HERG/EGFP) or to the N terminus (EGFP/HERG). These fusion constructs were transiently expressed in human embryonic kidney (HEK) 293 cells, and the whole-cell patch-clamp configuration and a confocal laser scanning microscope with primary anti-HERG antibodies and fluorescently labeled secondary antibodies were used. For EGFP/HERG channels the deactivation kinetics were faster and the peak tail current density was reduced when compared to both wild-type HERG channels and HERG/EGFP channels. Laser scanning microscopic studies showed that both fusion proteins were localized in the cytoplasm and on discrete microdomains in the plasma membrane. The extent of labeling with anti-HERG antibodies of HEK 293 cells expressing EGFP/HERG channels was less when compared to HERG/EGFP channels. In conclusion, both electrophysiological and immunocytochemical studies showed that EGFP/HERG channels themselves have a protein trafficking defect. HERG/EGFP channels have similar properties as untagged HERG channels and, thus, might be especially useful for fluorescence microscopy studies. PMID- 18414923 TI - A variant of the Ca2+-activated Cl channel Best3 is expressed in mouse exocrine glands. AB - Fluid secretion by exocrine glands requires the activation of an apical Ca2+ dependent Cl channel, the molecular identity of which is unknown. We found that mouse exocrine glands expressed an alternately spliced variant of Best3, a member of the Bestrophin (Vmd2) Ca2+-activated Cl channel gene family, whereas the heart expressed full-length Best3. The spliced transcript lacked exons 2, 3 and 6 (Best3-Delta2,3,6) and is predicted to generate an in-frame protein missing the entire cytoplasmic N terminus, the initial two transmembrane domains and part of the first intracellular loop. In addition to exocrine glands, the Best3 Delta2,3,6 splice variant transcript was detected in lung, testis and kidney. The parotid gland and heart expressed proteins of the predicted size for Best3 Delta2,3,6 and full-length Best3, respectively, that targeted to the plasma membrane in HEK293 cells. HEK293 cells expressing Best3 displayed Ca2+-dependent Cl(-) currents that were sensitive to the Cl channel blocker DIDS. In contrast, no Ca2+-dependent Cl(-) currents were detected in cells expressing Best3 Delta2,3,6. Cotransfection of Best3-Delta2,3,6 with Best3 or Best2 (also expressed in salivary gland acinar cells) had no significant effects on the currents generated by either of these Ca2+-dependent Cl channels. Our results demonstrate that exocrine glands express a unique splice variant of Best3. Nevertheless, Best3-Delta2,3,6 does not produce Ca2+-dependent Cl(-) currents, nor does it regulate the activity of Best2 or the full-length Best3 channel. PMID- 18414924 TI - Detection of the genes evolving under Ureaplasma-specific selection. AB - Mycoplasmas are parasitic bacteria with small genomes. Since parasitic bacteria need to adapt themselves to their hosts, there is a possibility that some genes evolved under species-specific constraint. We assume that Ureaplasma parvum has candidate genes that evolved in a species-specific manner in its genome. Here we examined synonymous-to-nonsynonymous substitution ratios (omega) of the 143 mycoplasma-orthologous genes of Ureaplasma and other mycoplasmas using branch models. As a result, the model allowing for Ureaplasma branch-specific omega in addition to omega of other mycoplasmas was significantly supported in 16 genes. First, the Ureaplasma-specific model was significantly supported in the genes encoding a transcription elongation factor and a transcription terminator factor, suggesting that transcription-related genes of Ureaplasma have evolved in a unique manner compared to those of other mycoplasmas. Second, the Ureaplasma specific model was significantly supported in the gene encoding uracil-DNA glycosylase. In addition, the omega value of the gene in the Ureaplasma lineage was approximately 30-fold lower than those of other lineages, suggesting that uracil-DNA glycosylase of Ureaplasma evolved under stronger functional constraint than those of other mycoplasmas. Finally, three glycolytic genes of Ureaplasma were suggested to have evolved under relaxed selection. Among mycoplasmas, only Ureaplasma has urease and synthesizes ATPs via hydrolysis of urea. This raises the possibility that Ureaplasma does not need a glycolysis pathway for ATP synthesis. This unique energy-producing system may be related to the Ureaplasma specific evolution of the glycolytic genes. PMID- 18414925 TI - Rapid evolution by positive Darwinian selection in T-cell antigen CD4 in primates. AB - CD4, an integral membrane glycoprotein, plays a critical role in the immune response and in the life cycle of simian and human immunodeficiency virus (SIV and HIV). Pairwise comparisons of orthologous human and mouse genes show that CD4 is evolving much faster than the majority of mammalian genes. The acceleration is too great to be attributed to a simple relaxation of the action of purifying selection alone. Here we show that the selective pressure acting on CD4 is highly variable between regions in the protein and identify codon sites under strong positive selection. We reconstruct the coding sequences for ancestral primate CD4s and model tertiary structures of all ancestral and extant sequences. Structural mapping of positively selected sites shows they distribute on the surface of the D1 domain of CD4, where the exogenous SIV gp120 protein binds. Moreover, structural models of the ancestral sequences show substantially larger variation in the interfacial electrostatic charge on CD4 and in the surface complementary between CD4 and gp120 in CD4 lineages from primates with natural SIV infections than those without. Thus, positive selection on CD4 among primates may reflect forces driven by SIV infection and could provide a link between changes in sequence and structure of CD4 during evolution and the interaction with the immunodeficiency virus. PMID- 18414926 TI - Evidence of gene conversion in the evolutionary process of the codon 41/42 ( CTTT) mutation causing beta-thalassemia in southern China. AB - The 4-bp deletion (-CTTT) at codon 41/42 (CD41/42) of the human beta-globin gene represents one of the most common beta-thalassemia mutations in East Asia and Southeast Asia, which is historically afflicted with endemic malaria, thus hypothetically evolving under natural selection by malaria infection. To understand the evolutionary process of generating the beta(CD41/42) allele and its maintenance, including the effect of natural selection on the pattern of linkage disequilibrium (LD), we sequenced a 15.933-kb region spanning 20.693 kb of the beta-globin cluster surrounding the 4-bp deletion using a sample from a Chinese population consisting of 24 normal individuals and 16 heterozygotes for the deletion. Forty-nine polymorphic sites were found. Analysis of the data, using a variety of methods including formal population genetics analysis and visual approaches, suggests that the spread of the CD41/42 (-CTTT) deletion is most likely mediated by interallelic gene conversion, although independent deletions in different lineages are also possible. The neutrality test resulted in a significant positive Tajima's D for the beta-globin locus, which is consistent with the existence of balancing selection. This suggests that the 4-bp deletion that occurred at this locus may be an event that is subject to natural selection, due to malaria, which leads to the heterozygote advantage, spread widely with further help by conversion and migration. The evolutionary process of this mutant through gene conversion that could conceivably take place between the 4-bp deletion and the normal sequence in the respective region is discussed in detail. PMID- 18414927 TI - Heavy metals and acidic components in total deposited matter in Sibenik and National Park Kornati, Croatia. AB - This article presents the results of 6 years (1999-2004) of monitoring acidic components (fluoride, chloride, nitrate, sulfate) and heavy metals (lead, cadmium, thallium) in total deposited matter (TDM) in the town of Sibenik and the National Park Kornati, Croatia, in order to determine their levels, spatial and temporal variations, and possible emission source. Bulk deposition samples were collected using the Bergerhoff samplers. TDM matter was determined gravimetrically. Acidic anions were analyzed using ion chromatography. Metallic components were determined by atomic absorption spectrometry. Results show that the levels of measured pollutants were relatively low compared to those reported in other investigations. Statistical analysis indicates that the measured pollutants originate from several sources such as resuspended soil, industry, road dust, traffic, and secondary aerosols as the most important. The levels of TDM show a slightly increasing trend at all sampling sites. A significant increasing trend in the levels of nitrate and sulfate in TDM could be the consequence of increasing traffic activity and urbanization. The significant increasing trend in the ratio between nitrate and sulfate in TDM indicates that the nitrate and sulfate relative contribution to acidification is increasing. The level of heavy metals in TDM was in order of lead > thallium > cadmium at all sampling sites with slightly decreasing trend. PMID- 18414928 TI - Estrogenic effects of selected hydroxy polychlorinated biphenyl congeners in primary culture of Atlantic Salmon (Salmo salar) hepatocytes. AB - Many persistent organic pollutants are known to have endocrine-disrupting effects in several aquatic and terrestrial species. In this regard, hydroxylated metabolites of polychlorinated biphenyls (OH-PCBs) represent serious health and environmental concern because they are shown to act agonistic or antagonistic at hormone receptors (HRs) or to cause hormone-receptor-mediated responses. In the present study, salmon primary hepatocytes were used to study alterations in an estrogen signaling pathway resulting from exposure to four hydroxylated (4OH-CB 107, 4OH-CB146, 4OH-CB187, and 3OH-CB138) metabolites of PCB at different concentrations using quantitative real-time polymerase chain reaction. The effects of the PCB metabolites were compared to the mRNA expression in 17alpha ethynylestradiol (EE2)-treated cells. Concentration-specific increase of vitellogenin (Vtg) mRNA transcription after exposure to OH-PCBs was observed. Decreased mRNA transcription was observed for zona radiata protein (Zr-protein) and cytochrome P450 side-chain cleavage (P450scc) enzyme. For estrogen receptor beta (ERbeta), the mRNA expression pattern was OH-PCB-metabolite congener specific. A novel aspect of this study is that OH-PCBs produced effects on hepatic steroidogenic pathways by targeting the StAR protein and P450scc genes. Given that endocrine toxicology research mainly has focused on estrogenicity involving direct ER-mediated effects and that steroidogenic enzyme and proteins are highly tissue- and cell-type-specific and controlled by different promoters and second-messenger pathways, the present findings provide potential new targets for interaction with xenobiotics such as hydroxylated congeners of certain chemicals. The quantitative expression patterns of hepatic and extrahepatic steroidogenic genes and proteins after exposure to environmental contaminants are the subject of systematic investigations in our laboratory. PMID- 18414929 TI - Mixed supra- and infradiaphragmatic totally anomalous pulmonary venous connection in an adolescent. AB - An unusual type of mixed totally anomalous pulmonary venous drainage in an 11 year-old boy was diagnosed on angiography. The left-side veins drained via a vertical vein into the portal venous system, while the right-side veins drained high into the superior vena cava. Surgical correction of this abnormality was successfully performed. PMID- 18414930 TI - Anomalous origin of left pulmonary artery from aorta with atrial septal defect. AB - Anomalous origin of left pulmonary artery (ALPA) from aorta with atrial septal defect (ASD) is a rare congenital cardiac malformation. We report a case of ALPA from the aorta with ASD in a 1.8-kg baby. Repair was needed at an early age due to progressive congestive heart failure. Surgical correction was performed with cardiopulmonary bypass for closure of ASD and direct anastomosis of the ALPA onto the main pulmonary artery. PMID- 18414931 TI - Successful use of bivalirudin for superior vena cava recanalization and stent placement in a child with heparin-induced thrombocytopenia. AB - Heparin-induced thrombocytopenia (HIT) is a potentially life-threatening, adverse effect of heparin therapy. Patients with this complication require an alternative approach to anticoagulation. Bivalirudin is a direct thrombin inhibitor with an efficacy comparable to that of heparin, a short half-life, and reduced bleeding complications in adults. We present the case of a 2-year-old boy with HIT Type II who underwent recanalization of an occluded superior vena cava and stent placement, utilizing bivalirudin as anticoagulant. PMID- 18414932 TI - Small bowel involvement in blue rubber bleb nevus syndrome: MR imaging features. AB - Blue rubber bleb nevus syndrome (BRBNS) is a rare condition which is characterized by multiple venous malformations that may affect several organs. Gastrointestinal lesions, which mostly involve the small bowel and distal colon, may be responsible for gastrointestinal bleeding, causing more frequently chronic anemia. We report herein two cases of BRBNS causing gastrointestinal bleeding which were investigated by means of MR imaging. All venous malformations showed markedly high signal intensity on fat-suppressed T2-weighted MR images. In addition, extraintestinal venous malformations displayed high signal intensity similar to that of small bowel venous malformations. Fat-suppressed T2-weighted TSE images show both small bowel and colonic involvement, thus providing useful information relative to the most appropriate therapeutic approach to be used. MR imaging is especially helpful when capsule endoscopy or enteroscopy are contraindicated or not feasible. In addition, because of lack of radiation, MR imaging can be performed repeatedly to assess the evolution of the disease before a planned surgery or endoscopic resection. Also, because BRBNS is by essence a multifocal condition with extraintestinal involvement, MR imaging appears as an appropriate imaging technique to investigate the multiple locations of the disease in one session, thus replacing multiple examinations. PMID- 18414934 TI - Flat polyps of the colon: accuracy of detection by CT colonography and histologic significance. AB - There is controversy regarding the prevalence, clinical importance, and appropriate screening methods for nonpolypoid (flat and depressed) polyps in the colon. Investigators in Japan have reported higher prevalence of nonpolypoid adenomas in the general population and there have been several reports of higher incidence of high-grade dysplasia in flat adenomas in these Eastern studies. Historically, many Western gastroenterologists have been skeptical of these findings and there have been conflicting studies regarding the prevalence of flat adenomas and incidence of high-grade dysplasia in these lesions. Multiple reasons have been postulated for this apparent difference. Therefore further research into this topic is needed to clarify these issues. In this article we will review the controversy related to the definitions and clinical importance of nonpolypoid neoplasms in the colon, demonstrate the appearance of these unique lesions at CT colonography (CTC) and discuss the accuracy of CTC. PMID- 18414935 TI - Comparing cephaloauricular and scaphaconchal angles in prominent ear patients and control subjects. AB - Approximately 5% of 1-year-old children have prominent ears. The most common findings are underdevelopment or lack of the antihelical fold, overdevelopment of the concha, a scapha-conchal angle greater than 130 degrees, and a protruding lobule. This study compared the cephaloauricular and scaphaconchal angles of 15 patients with prominent ears and 15 patients in a control group. Alginate was used to create a mold of each patient's right ear. Afterward the molds were cut transversally for measurement of the angles. The first cut was made at the middle of the ear's cephalocaudal length. The second cut was made in the superior piece midway between the first cut and the superior extremity of the ear. The cephaloauricular angle was defined as the intersection of a straight line running through the tragus insertion and the lateral portion of the mastoid region with a straight line that running through the tragus and the middle of the helix. The scaphaconchal angle was obtained in the second cut by measurement of the angles formed by these two structures molded in the posterior aspect of the ear. The Student's t test was used for statistical analysis. The average cephaloauricular angle was 47.7 degrees for the study group and 31.1 degrees for the control group. The average scaphaconchal angle was 132.6 degrees for the study group and 106.7 degrees for the control group. This study presents a new method for evaluating the angles of the ear, confirming that both measured angles (cephaloauricular and scaphaconchal) are greater in patients with prominent ears (p < 0.005). PMID- 18414936 TI - Human adipose-derived stem cells adhere to acellular dermal matrix. PMID- 18414937 TI - A tie-over dressing for the ear skin graft. PMID- 18414938 TI - Hemostasis in skin surgery. PMID- 18414939 TI - Land-use planning in the Chaco plain (Burruyacu, Argentina): part 2: generating a consensus plan to mitigate land-use conflicts and minimize land degradation. AB - The Burruyacu district (Tucuman province, Northwest Argentina) has been traditionally an area with rural activities based on the exploitation of the Chaco forest for timber and livestock browsing. Since the 1960s, local institutions started promoting soybean due to favorable land conditions and good market prices. Soybean extension, as from the 1970s, has resulted in important reduction of the Chaco forest and also caused physical soil degradation, especially soil compaction and erosion. A land-use-planning exercise was carried out using the Land-Use Planning and Information System (LUPIS) as a spatial decision support system. LUPIS facilitates the generation of alternative land-use plans by adjusting the relative importance attributed by multiple stakeholders to preference and avoidance policies. The system leads to the allocation of competing land uses to land map units in accordance with their preferred resource requirements, conditional upon the resource base of the area and the stakeholders' demands. After generating a land use plan for each stakeholder category identified in the study area, including commercial farmers, conservative/conventional farmers, and conservationists, a consensus plan was established to address the land-use conflicts between mechanized agriculture, traditional agriculture and forest conservation, and to mitigate soil degradation caused by extensive dry-farming. Although the planning exercise did not directly involve the stakeholders, the results are sufficiently practical and realistic to suggest that the approach could be extended to the entire Chaco plain region. PMID- 18414940 TI - Users' perception as a tool to improve urban beach planning and management. AB - Four beaches that share physiographic characteristics (sandy, wide, and long) but differ in socioeconomic and cultural terms (three are located in northwestern Mexico and one in California, USA) were evaluated by beach users. Surveys (565) composed of 36 questions were handed out to beach users on weekends and holidays in 2005. The 25 questions that revealed the most information were selected by factor analysis and classified by cluster analysis. Beach users' preferences were assigned a value by comparing the present survey results with the characteristics of an "ideal" recreational urban beach. Cluster analysis separated three groups of questions: (a) services and infrastructure, (b) recreational activities, and (c) beach conditions. Cluster linkage distance (r=0.82, r=0.78, r=0.67) was used as a weight and multiplied by the value of beach descriptive factors. Mazatlan and Oceanside obtained the highest values because there are enough infrastructure and services; on the contrary, Ensenada and Rosarito were rated medium and low because infrastructure and services are lacking. The presently proposed method can contribute to improving current beach evaluations because the final score represents the beach users' evaluation of the quality of the beach. The weight considered in the present study marks the beach users' preferences among the studied beaches. Adding this weight to beach evaluation will contribute to more specific beach planning in which users' perception is considered. PMID- 18414941 TI - Spatial analysis of soil salinity and soil structural stability in a semiarid region of New South Wales, Australia. AB - Salt-affected soils are a major threat to agriculture especially in the semiarid regions of the world. The effective management of these soils requires adequate understanding of not only how water and, hence, solutes are transported within the soil, but also how soil salinity and sodicity spatially interact to determine soil structural breakdown. For sustainable agricultural production, information on quantitative soil quality, such as salinity, is required for effective land management and environmental planning. In this study, quantitative methods for mapping indicators of soil structural stability, namely salinity and sodicity, were developed to assess the effect of these primary indicators on soil structural breakdown. The current levels of soil salinity, as measured by electrical conductivity (EC) of the soil/water suspension, soil sodicity, represented by exchangeable sodium percentage (ESP), and aggregate stability, were assessed. Remote sensing, geographical information system (GIS), and geostatistical techniques--primarily regression-kriging and indicator-kriging- were used to spatially predict the soil sodicity and salinity. The patterns of salinity (EC) and sodicity (ESP>5%) were identified. The effect of land use on these soil quality indicators was found to be minimal. Co-spatial patterns were elucidated between sodic soils (defined by ESP>5%) and highly probable mechanically dispersive soils predicted from indicator-kriging of ASWAT scores. It was established that the incorporation of EC with ESP into an objective index, called electrolyte stability index (ESI=ESP/EC), gave a good indication of soil dispersion, although the threshold ESI value below which effective structural breakdown might occur is 0.025, which is twice as small as the expected 0.05. The discrepancies between ESI and ASWAT scores suggest that other soil factors than salinity and sodicity are affecting soil structural breakdown. This calls for further investigation. The study provides valuable information in the form of risk zones of soil structural breakdown for land management. These zones, with a probability of mechanical soil dispersion of >0.70, require immediate management attention with greater monitoring and amelioration techniques, particularly gypsum or lime application and/or altered cultivation techniques. PMID- 18414942 TI - What is a surgical complication? PMID- 18414944 TI - Endovascular management of ruptured pancreaticoduodenal artery aneurysms associated with celiac axis stenosis. AB - The purpose of this study was to assess the efficacy of transcatheter arterial embolization for ruptured pancreaticoduodenal artery (PDA) aneurysms associated with celiac axis stenosis (CS). Seven patients (four men and three women; mean age, 64; range, 43-84) were treated with transcatheter arterial embolization between 2002 and 2007. They were analyzed with regard to the clinical presentation, radiological finding, procedure, and outcome. All patients presented with sudden epigastric pain or abdominal discomfort. Contrast-enhanced CT showed a small aneurysm and retroperitoneal hematoma around the pancreatic head in all patients. The aneurysms ranged from 0.3 to 0.9 cm in size. In one patient, two aneurysms were detected. The aneurysms were located in the pancreaticoduodenal artery (n = 5) and the dorsal pancreatic artery (n = 3). Embolization was performed with microcoils in all aneurysms (n = 8). N-Butyl 2 cyanoacrylate (n = 1) and gelatine particle (n = 1) were also used. Complete occlusion was achieved in four patients. In the other three patients, a significantly reduced flow to the aneurysm remained at final angiography. However, these aneurysms were thrombosed on follow-up CT within 2 weeks. And there was no recurrence of the symptoms and bleeding during follow-up (mean, 28 months; range, 5-65 months) in all patients. In conclusion, transcatheter arterial embolization for PDA aneurysms associated with CS is effective. Significant reduction of the flow to the aneurysm at final angiography may be predictive of future thrombosis. PMID- 18414945 TI - Femoral artery stenosis following percutaneous closure using a starclose closure device. AB - Starclose (Abbott Vascular Devices, Redwood City, CA) is a new arterial closure device that seals a femoral puncture site with an extravascular star-shaped nitinol clip. The clip projects small tines into the arterial wall which fold inward, causing the arterial wall to pucker, producing a purse-string-like seal closing the puncture site. The case history is that of a 76-year-old female patient who underwent day-case percutaneous diagnostic coronary angiography. A Starclose femoral artery closure device was used to achieve hemostasis with subsequent femoral artery stenosis. PMID- 18414946 TI - Subintimal angioplasty for peripheral arterial occlusive disease: a systematic review. AB - The objective of this study was to summarize outcomes of subintimal angioplasty (SA) for peripheral arterial occlusive disease. The Cochrane Library, Medline and Embase databases were searched to perform a systematic review of the literature from 1966 through May 2007 on outcomes of SA for peripheral arterial occlusive disease of the infrainguinal vessels. The keywords "percutaneous intentional extraluminal revascularization," "subintimal angioplasty," "peripheral arterial disease," "femoral artery," "popliteal artery," and "tibial artery" were used. Assessment of study quality was done using a form based on a checklist of the Dutch Cochrane Centre. The recorded outcomes were technical and clinical success, primary (assisted) patency, limb salvage, complications, and survival, in relation to the clinical grade of disease (intermittent claudication or critical limb ischemia [CLI] or mixed) and location of lesion (femoropopliteal, crural, or mixed). Twenty-three cohort studies including a total of 1549 patients (range, 27 to 148) were included in this review. Methodological and reporting quality were moderate, e.g., there was selection bias and reporting was not done according to the reporting standards. These and significant clinical heterogeneity obstructed a meta-analysis. Reports about length of the lesion and TASC classification were too various to summarize or were not mentioned at all. The technical success rates varied between 80% and 90%, with lower rates for crural lesions compared with femoral lesions. Complication rates ranged between 8% and 17% and most complications were minor. After 1 year, clinical success was between 50% and 70%, primary patency was around 50% and limb salvage varied from 80% to 90%. In conclusion, taking into account the methodological shortcomings of the included studies, SA can play an important role in the treatment of peripheral arterial disease, especially in the case of critical limb ischemia. Despite the moderate patency rates after one year, SA may serve as a "temporary bypass" to provide wound healing and limb salvage. PMID- 18414948 TI - NOTES: Where have we been and where are we going? PMID- 18414949 TI - Olfactory responses of the predatory mites (N eoseiulus cucumeris) and insects (Orius strigicollis ): to two different plant species infested with onion thrips (T hrips tabaci). AB - Responses of Neoseiulus cucumeris (a predatory mite) and the predatory insect Orius strigicollis to volatiles associated with two different plant species infested with onion thrips, Thrips tabaci, were examined in a Y-tube olfactometer. Both predators species showed a significant preference for volatiles from infested cucumber leaves without T. tabaci over clean air. However, they were not attracted to volatiles from uninfested cucumber leaves, artificially damaged cucumber leaves, or volatiles from T. tabaci plus their visible products collected from cucumber leaves. These results suggest that both predator species are capable of exploiting herbivore-induced volatiles from T. tabaci-infested cucumber leaves as a foraging cue. Neither predator was attracted to volatiles from uninfested spring onion leaves, infested spring onion leaves without T. tabaci, or volatiles from T. tabaci plus their visible products collected from spring onion leaves. Interestingly, they avoided volatiles from artificially damaged spring onion leaves. A possible explanation for the non significant olfactory responses of the predator species to spring onion plants with infestation damage of T. tabaci is discussed. PMID- 18414950 TI - The iridoid glucoside, antirrhinoside, from Antirrhinum majus L. has differential effects on two generalist insect herbivores. AB - The iridoid glucoside, antirrhinoside, is constitutively distributed throughout Antirrhinum majus L. in a manner consistent with its possible role as an allelochemical, but there is no evidence that it has a defensive function with respect to insect herbivory. To address this question, two generalist herbivores, Lymantria dispar L. (gypsy moth) and Trichoplusia ni Hubner (cabbage looper) were chosen for feeding trials on excised whole leaves of A. majus and in artificial diet assays. In leaf excision feeding trials, fourth instar gypsy moth rejected, without sampling, the leaves of A. majus regardless of what node the leaf was excised from. In contrast, fourth instar cabbage looper readily fed on the excised leaves, and antirrhinoside was not found in their bodies or feces (frass) as determined by thin layer and high-pressure liquid chromatography. In the leaf and diet assays, a second major leaf iridoid in A. majus, antirrhide, was found in both cabbage looper and gypsy moth frass. In diet feeding assays, the growth of gypsy moth and cabbage looper were not inhibited by methanol extracts, iridoid fractions, or pure antirrhinoside at concentrations of 0.6% in diet, but cabbage looper growth was enhanced. At an antirrhinoside concentration of 3.3% in diet, gypsy moth growth was reduced, whereas cabbage looper growth again increased significantly relative to the control. It is likely that antirrhinoside functions as defense against herbivory for one generalist insect herbivore but also, at low concentrations, enhances the growth of another. PMID- 18414951 TI - Squamous cell carcinoma of the anal canal: utilization and outcomes of recommended treatment in the United States. AB - BACKGROUND: Over the past two decades, recommended treatment for squamous cell carcinoma of the anal canal has shifted from surgery to primary chemoradiation. Resection is now reserved for persistent or recurrent disease. Our objectives were (1) to evaluate treatment trends over the past 20 years, (2) to assess contemporary treatment utilization, and (3) to examine the impact of recommended vs nonguideline treatment on survival. METHODS: From the National Cancer Data Base (1985-2005), 38,882 patients with anal canal cancer were identified. Regression models were used to assess factors associated with use of nonguideline treatment (vs chemoradiation +/-surgery). Univariate and multivariate methods were used to assess the impact of treatment on survival. RESULTS: From 1985 to 2005, the use of chemoradiation increased significantly with a concomitant decrease in treatment with surgery alone (P < .0001). However, only 74.9% (5014 of 6696) of patients underwent primary chemoradiation therapy in 2003-2005. Overall, 22.7% (1523 of 6696) of patients received treatment that was not concordant with established guidelines: primary surgery (13.0%) and primary chemotherapy or radiation (9.7%). Patients were significantly less likely to receive guideline treatment if male, older, black or Hispanic, more severe comorbidities, or Stage I (vs Stage II or III). Patients undergoing chemoradiation ( +/- surgery) had higher 5-year survival rates than patients who received nonguideline treatment (64% vs 58%; hazard ratio 0.82, 95% confidence interval [95% CI] 0.77-0.87; P < .0001). CONCLUSION: Primary chemoradiation therapy has supplanted surgical treatment and is associated with better outcomes; however, nearly a quarter of patients are still receiving treatment that is not concordant with established guidelines. PMID- 18414952 TI - Complete lymph node dissection for sentinel node-positive melanoma: assessment of practice patterns in the United States. AB - BACKGROUND: Currently, complete lymph node dissection (CLND) is recommended after identification of a metastatic lymph node by sentinel lymph node biopsy (SLNB). Guidelines suggest that CLND should be performed as a separate procedure, and a sufficient number of nodes should be examined. Our objective was to examine the utilization, timing, and adequacy of CLND for melanoma in the United States. METHODS: From the National Cancer Data Base, patients diagnosed with stage I to III melanoma during 2004-2005 were identified. Multiple logistic regression was used to assess factors associated with CLND utilization, timing (separate operation from SLNB), and adequacy (examination of > or = 10 nodes). RESULTS: Of the 44,548 patients identified, 47.5% were pathologic stage IA, 23.8% stage IB, 14.1% stage II, and 14.6% stage III. Of the 17% (2942 of 17,524) with nodal metastases on SLNB, only 50% underwent a CLND. Patients were significantly less likely to undergo a CLND after SLNB if > 75 years old or had lower extremity melanomas. Of the patients who underwent a CLND, only 42% underwent the CLND at a separate procedure after the SLNB. Of those who underwent a CLND, 69.2% had > or = 10 nodes examined. Patients were significantly less likely to have > or = 10 nodes examined if they were > 75 years old or had lower extremity melanomas. Patients treated at NCCN/NCI-designated centers were significantly more likely to undergo nodal evaluation in concordance with established guidelines. CONCLUSIONS: Only half of patients with sentinel node-positive melanoma underwent CLND. Quality surveillance measures are needed to monitor, standardize, and improve the care of patients with malignant melanoma. PMID- 18414953 TI - Experiences with hospital care: perspectives of black and Hispanic patients. AB - BACKGROUND: Significant racial and ethnic differences along several dimensions of patients'experiences with hospital care have been previously documented. However, the relationship between these differences and possible differences in processes of care has not been well described. METHODS: We conducted focus groups with 37 black and Hispanic men and women who had recently been discharged from either medical or obstetrical services at an urban academic medical center to assess which dimensions of these patients' experiences with care were most important in determining overall levels of satisfaction. RESULTS: Differences were found between Hispanics and blacks in the factors that influence their overall positive and negative experiences. Participants identified two themes that influence experiences with hospital care that are not commonly examined in many patient satisfaction instruments: availability and quality of translators, and attitudes of social workers and nursing staff. CONCLUSIONS: Our findings suggest that hospitals should pursue hiring a culturally diverse work force and should collect racial and ethnically specific data about satisfaction with care including satisfaction with availability of social workers and interpreters. PMID- 18414954 TI - Clinical utility of folic acid testing for patients with anemia or dementia. AB - BACKGROUND: Serum folic acid tests are routinely ordered by physicians for evaluating anemia and sometimes ordered for evaluating dementia and altered mental status. OBJECTIVE: To determine the utility of routine folic acid testing for patients with anemia or dementia/altered mental status in the era of folic acid fortification. DESIGN: Retrospective analysis of consecutive folic acid tests performed on adults over a 4-month period; chart review of patients without anemia. MEASUREMENTS AND MAIN RESULTS: Serum folic acid level, mean corpuscular volume (MCV), and hematocrit. We reviewed 1,007 folic acid tests performed on 980 patients. The average age was 63.8 years, and 62% of the tests were from outpatient facilities. Only 4 (0.4%) patients had folic acid levels <3 ng/mL, while 10 (1%) patients had levels of 3-4 ng/mL (borderline). Thirty-five percent of the folic acid tests were performed on patients who were not anemic; most of these were ordered to evaluate dementia or altered mental status and folic acid level was normal in all these patients. Only 7% of the patients tested had a macrocytic anemia; these patients were more likely than those without macrocytic anemia to have low folic acid levels (2.8% vs 0.4%, p < .03). CONCLUSION: Low serum folic acid levels were rarely detected in a series of patients being evaluated for anemia, dementia, or altered mental status. The test should be reserved for patients with macrocytic anemia and those at high risk for folic acid deficiency. PMID- 18414955 TI - Learning from the legal history of billing for medical fees. AB - INTRODUCTION: When patients pay for care out-of-pocket, physicians must balance their professional obligations to serve with the commercial demands of medical practice. Consumer-directed health care makes this problem newly pressing, but law and ethics have thought for millennia about how doctors should bill patients. HISTORICAL BACKGROUND: At various points in European history, the law restricted doctors' ability to bill for their services, but this legal aversion to commercializing medicine did not take root in the American colonies. Rather, US law has always treated selling medical services the way it treats other sales. Yet doctors acted differently in a crucial way. Driven by the economics of medical practice before the spread of health insurance, doctors charged patients according to what they thought each patient could afford. The use of sliding fee scales persisted until widespread health insurance drove a standardization of fees. CURRENT PRACTICE: Today, encouraged by Medicare rules and managed care discounts, providers use a perverse form of a sliding scale that charges the most to patients who can afford the least. Primary care physicians typically charge uninsured patients one third to one half more than they receive from insurers for basic office or hospital visits, and markups are substantially higher (2 to 2.5 times) for high-tech tests and specialists' invasive procedures. CONCLUSION: Ethical and professional principles might require providers to return to discounting fees for patients in straitened circumstances, but imposing such a duty formally (by law or by ethical code) on doctors would be harder both in principle and in practice than to impose such a duty on hospitals. Still, professional ethics should encourage physicians to give patients in economic trouble at least the benefit of the lowest rate they accept from an established payer. PMID- 18414956 TI - Effects of military trauma exposure on women veterans' use and perceptions of Veterans Health Administration care. AB - BACKGROUND: Few studies have addressed how military trauma exposure, particularly sexual assault and combat exposure, affects women veterans' use and perceptions of Veterans Health Administration (VHA) care. OBJECTIVE: The aim of the present study was to evaluate the effects of military sexual assault and combat exposure on women veterans' use and perceptions of different aspects of VHA care. DESIGN: Cross-sectional telephone survey of a national sample of women veterans. PARTICIPANTS: Women from the VA's National Registry of Women Veterans. MEASUREMENTS: Sociodemographic characteristics, VHA care utilization, perceptions of care. RESULTS: Women veterans with histories of military sexual assault reported more use of VHA services, but less satisfaction, poorer perceptions of VHA facilities and staff, and more problems with VHA services compared to women veterans without histories of sexual assault. Combat exposure was related to more problems with VHA staff, although few other differences were observed for women with and without histories of combat exposure. CONCLUSIONS: Findings provide information on areas that can be targeted with respect to caring for women veterans exposed to military sexual trauma and combat exposure, including improving interactions with VHA staff and the ease of using VHA services. PMID- 18414957 TI - Band and port-related morbidity after bariatric surgery: an underestimated problem. AB - OBJECTIVE: To evaluate the magnitude of the morbidity related to the system used for gastric banding Methods Between January 1997 and December 2004, 286 consecutive patients underwent laparoscopic gastric banding (LAGB) in one center. We used 4 models of LapBand 9.75, 10, 11 and Vanguard with pars flacida route. Recalibration of band was performed in our consultation unit without systematic radiologic control. We considered four kinds of complication: port displacement, port rupture, band rupture and others problems. RESULTS: The mean follow up was 3.3+/-2.8 years with a median 2.9 years. Complications occurred within a mean time of 2.2+/-1.9 years. For the models vanguard and size 11, there were no rupture and 15 (27.7%) displacements whereas for size 9.75 and 10 there were 39 ruptures (14.7%) and 15 (5.6%) displacements. Types of complications were related to the bands used i.e. more port displacements for the models vanguard and size 11 and more band and port ruptures for the models size 9.75 and 10. But when we considered the respective follow up according to the type of band these differences were no longer significant. Moreover rupture rate was significantly high but decreased after March 2002 because of changing of junction between port and catheter. Mean excess weight loss (35.2+/-27.7%) was not different in group whether the patients were reoperated or not. CONCLUSION: Band and port related morbidity is an important aspect of bariatric surgery. We have to pay attention to material evolution and to our follow up for calibration. Some new recent technical advancement could improve the management of these patients. PMID- 18414958 TI - Leukocyte scintigraphy SPECT/CT software fusion diagnosis of infected gastric band. AB - The prevalence of morbid obesity is increasing worldwide and presents a significant clinical problem. Laparoscopic adjustable gastric banding has been popularized as an effective, safe, minimally invasive surgical technique for the treatment of morbid obesity. Different diagnostic imaging modalities are crucial for follow-up of "banded" patients and detection of the different postoperative complications. We present a case report including detailed clinical, laboratory, radiological, and scintigraphic findings in a 50-year-old woman with an infected gastric band diagnosed by technetium 99m-hexamethyl propilenamine oxime-labeled leukocyte scintigraphy and discuss the value of integrated interpretation of anatomical and functional imaging modalities obtained by software fusion technique. PMID- 18414959 TI - Continuous hemodynamic monitoring during laparoscopic gastric bypass in superobese patients by pressure recording analytical method. AB - BACKGROUND: Morbid obesity, compromising cardiovascular and respiratory function, may increase the risk of anesthesia and was initially considered a contraindication to laparoscopy. The aim of this study was to investigate hemodynamic effects induced by pneumoperitoneum in superobese patients, assessed by arterial pulse contour method. METHODS: We prospectively studied 10 obese patients (BMI 53 +/- 9 kg/m(2)), scheduled for laparoscopic gastric bypass. After anesthesia induction, patients were intubated and mechanically ventilated. A radial artery was cannulated to obtain hemodynamic data implemented by means of a new pulse contour analysis method-the pressure recording analytical method (PRAM). Data were recorded after anesthesia induction (Tbas), at peritoneal insufflation (T0), at 1, 3, 5, 10, 30, and 60 min after pneumoperitoneum induction (T1, T3, T5, T10, T15, T30, T60), at abdominal desufflation (Tdef) and 5 min after desufflation (T5def). RESULTS: MAP increased after pneumoperitoneum, returning to its baseline after deflation (79 +/- 7 Tbas, 81 +/- 6 mmHg T5def). HR remained unchanged. Systemic vascular resistance index (SVRI) increased after pneumoperitoneum induction and progressively returned to baseline (3,903 +/- 330 Tbas, 4,596 +/- 148 T1, and 3,786 +/- 202 dyn s m(2) cm(-5) T5def). Stroke volume index (SVI) and cardiac index (CI) increased after pneumoperitoneum and remained elevated. Stroke volume variation (SVV) decreased after insufflation remaining lower than the basal value (28 +/- 4% Tbas, 15 +/- 5% T5des). Cardiac Cycle Efficiency (CCE) transient decreased after insufflation and subsequently increased (0.029 +/- 0.146 Tbas, 0.008 +/- 0.178 T5def). Aortic dP/dt max increased after insufflation, returning to baseline only after desufflation (0.68 +/- 0.07 Tbas, 0.94 +/- 0.08 T30 and 0.86 +/- 0.06 mmHg s(-1) T5def). CONCLUSION: As assessed by PRAM, this study showed no deterioration in hemodynamic indices or ventricular performance during laparoscopic gastric bypass. PMID- 18414960 TI - Emergency sleeve gastrectomy as rescue treatment for acute gastric necrosis due to type II paraesophageal hernia in an obese woman with gastric banding. AB - A morbidly obese 42-year-old woman presented with a 1-week history of left chest pain. She had undergone laparoscopic adjustable gastric banding 16 months earlier with a body mass index (BMI) of 49.2 kg/m2. Diagnostic workup revealed a large left pleural empyema and ruled out band slippage. At left thoracotomy, a misdiagnosed type II paraesophageal strangulated hernia with gastric necrosis and large perforation of the fundus was evident. At laparotomy, the band was removed, the stomach was reduced into the abdomen, and a sleeve gastrectomy was performed. Her postoperative course was uneventful, and 6 months after surgery, her BMI is 31 kg/m2. Emergency sleeve gastrectomy could represent a good option to treat, at the same time and in a safe way, both gastric necrosis and paraesophageal hernia, improving the good results in terms of weight loss after gastric restriction from gastric banding. PMID- 18414961 TI - New tendon transfer for correction of drop-foot in common peroneal nerve palsy. AB - Common peroneal nerve palsy has been reported to be the most frequent lower extremity palsy characterized by a supinated equinovarus foot deformity and foot drop. Dynamic tendon transposition represents the gold standard for surgical restoration of dorsiflexion of a permanently paralyzed foot. Between 1998 and 2005, we operated on a selected series of 16 patients with traumatic complete common peroneal nerve palsy. In all cases, we performed a double tendon transfer through the interosseous membrane. The posterior tibialis tendon was transferred to the tibialis anterior rerouted through a new insertion on the third cuneiform and the flexor digitorum longus was transferred to the extensor digitorum longus and extensor hallucis longus tendons. All 16 patients were reviewed at a minimum followup of 24 months (mean, 65 months; range, 24-114 months). The results were assessed using the Stanmore system questionnaire and were classified as excellent in eight, good in five, fair in two, and poor in one. Postoperative static and dynamic baropodometric evaluations also were performed. The proposed procedure, which provides an appropriate direction of pull with adequate length and fixation, is a reliable new method to restore balanced foot dorsiflexion correcting the foot and digit drop and producing a normal gait without the use of orthoses. LEVEL OF EVIDENCE: Level IV, therapeutic study. See the Guidelines for Authors for a complete description of levels of evidence. PMID- 18414962 TI - Lifetime characteristics of participants and non-participants in a smoking cessation trial: implications for external validity and public health impact. AB - BACKGROUND: Detailed information about the characteristics of smokers who do and do not participate in smoking cessation treatment is needed to improve efforts to reach, motivate, and treat smokers. PURPOSE: The aim of this study is to explore a broad range of characteristics related to participation in a smoking cessation trial. METHODS: Eligible smokers were recruited from a longitudinal birth cohort. Participants and non-participants were compared on a broad range of sociodemographics, smoking, psychiatric and substance abuse disorders, personality, and prospective measures from early childhood. Eligible smokers were compared to a matched regional subsample of the Behavioral Risk Factor Surveillance System (BRFSS). RESULTS: Few differences were observed, most of which were statistically significant but not clinically meaningful. Compared to non-participants, participants were more likely to be single, have lower income, be more nicotine-dependent, be more motivated to quit, and have higher levels of depressed mood and stress even after covariance of gender, income, and marital status. Sociodemographic differences between participants and the BRFSS sample reflect the skew toward lower socioeconomic status in the original birth cohort. CONCLUSIONS: The encouraging conclusion is that smokers who enroll in cessation trials may not differ much from non-participants. Information about treatment participants can inform the development of recruitment strategies, improve the tailoring of treatment to individual smoker profiles, help to estimate potential selection bias, and improve estimates of population impact. PMID- 18414963 TI - Genetic influences on bone loss in the San Antonio Family Osteoporosis study. AB - The genetic contribution to age-related bone loss is not well understood. We estimated that genes accounted for 25-45% of variation in 5-year change in bone mineral density in men and women. An autosome-wide linkage scan yielded no significant evidence for chromosomal regions implicated in bone loss. INTRODUCTION: The contribution of genetics to acquisition of peak bone mass is well documented, but little is known about the influence of genes on subsequent bone loss with age. We therefore measured 5-year change in bone mineral density (BMD) in 300 Mexican Americans (>45 years of age) from the San Antonio Family Osteoporosis Study to identify genetic factors influencing bone loss. METHODS: Annualized change in BMD was calculated from measurements taken 5.5 years apart. Heritability (h(2)) of BMD change was estimated using variance components methods and autosome-wide linkage analysis was carried out using 460 microsatellite markers at a mean 7.6 cM interval density. RESULTS: Rate of BMD change was heritable at the forearm (h(2) = 0.31, p = 0.021), hip (h(2) = 0.44, p = 0.017), spine (h(2) = 0.42, p = 0.005), but not whole body (h(2) = 0.18, p = 0.123). Covariates associated with rapid bone loss (advanced age, baseline BMD, female sex, low baseline weight, postmenopausal status, and interim weight loss) accounted for 10% to 28% of trait variation. No significant evidence of linkage was observed at any skeletal site. CONCLUSIONS: This is one of the first studies to report significant heritability of BMD change for weight-bearing and non weight-bearing bones in an unselected population and the first linkage scan for change in BMD. PMID- 18414964 TI - The BPAQ: a bone-specific physical activity assessment instrument. AB - A newly developed bone-specific physical activity questionnaire (BPAQ) was compared with other common measures of physical activity for its ability to predict parameters of bone strength in healthy, young adults. The BPAQ predicted indices of bone strength at clinically relevant sites in both men and women, while other measures did not. INTRODUCTION: Only certain types of physical activity (PA) are notably osteogenic. Most methods to quantify levels of PA fail to account for bone relevant loading. Our aim was to examine the ability of several methods of PA assessment and a new bone-specific measure to predict parameters of bone strength in healthy adults. METHODS: We recruited 40 men and women (mean age 24.5). Subjects completed the modifiable activity questionnaire, Bouchard 3-day activity record, a recently published bone loading history questionnaire (BLHQ), and wore a pedometer for 14 days. We also administered our bone-specific physical activity questionnaire (BPAQ). Calcaneal broadband ultrasound attenuation (BUA) (QUS-2, Quidel) and densitometric measures (XR-36, Norland) were examined. Multiple regression and correlation analyses were performed on the data. RESULTS: The current activity component of BPAQ was a significant predictor of variance in femoral neck bone mineral density (BMD), lumbar spine BMD, and whole body BMD (R(2) = 0.36-0.68, p < 0.01) for men, while the past activity component of BPAQ predicted calcaneal BUA (R(2) = 0.48, p = 0.001) for women. CONCLUSIONS: The BPAQ predicted indices of bone strength at skeletal sites at risk of osteoporotic fracture while other PA measurement tools did not. PMID- 18414965 TI - The place of methotrexate perioperatively in elective orthopedic surgeries in patients with rheumatoid arthritis. AB - No clear consensus exists on whether methotrexate (MTX) should be continued or whether this therapy should be discontinued for a few weeks in patients with rheumatoid arthritis (RA) undergoing surgery. Continued MTX therapy may impair wound healing, but discontinuation of the therapy may increase the risk of flares. In this article we review published data on the perioperative management of MTX in patients with RA undergoing elective orthopedic surgery. Eight papers on this topic could be identified. These studies compare continued vs. discontinued MTX therapy or MTX therapy vs. therapies other than MTX. Summing up the published data, continued MTX therapy appears to be safe perioperatively and seems also to be associated with a reduced risk of flares. None of the examined papers addresses the issue of safety in connection with comorbidities, age or high doses of MTX. PMID- 18414966 TI - Serum levels of natriuretic peptides in patients with Behcet's disease. AB - The objective of our study was to elucidate serum levels of atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP), and C-type natriuretic peptide (CNP) in Behcet's disease (BD) patients with active and inactive period. The multicenter study included 53 patients with active (n = 28) and inactive (n = 25) BD (mean age, 34.3 +/- 9 years; 15 men and 38 women) satisfying the International Study Group criteria and 26 healthy controls (mean age, 34.4 +/- 6.1 years; seven men and 19 women) matched for age and gender from a similar ethnic background. Serum natriuretic peptides levels were determined by enzyme immunoassay kit. Mean serum ANP concentrations in the active patients (4.01 +/- 1.21 ng/ml) were significantly lower than in the healthy controls (5.76 +/- 1.99 ng/ml, p = 0.004). Mean serum BNP levels were found to be significantly higher in both the active (6.19 +/- 2.97 ng/ml) and inactive (6.49 +/- 2.88 ng/ml) BD groups compared with the control group (3.82 +/- 1.1 ng/ml, p = 0.004 and p = 0.001, respectively). Mean serum CNP concentrations in the active patients (0.49 +/- 0.12 ng/ml) were significantly lower than in the inactive patients (0.65 +/- 0.2 ng/ml, p = 0.017) and the healthy controls (0.8 +/- 0.27 ng/ml, p < 0.001). Our results suggest that changes in natriuretic peptide levels may be associated with vasculitis that play role in the etiopathogenesis of the BD. PMID- 18414967 TI - Clinical impact of switching from infliximab to etanercept in patients with rheumatoid arthritis. AB - We assessed the disease activity in patients with rheumatoid arthritis (RA) after switching from infliximab to etanercept according to the reason of infliximab discontinuation. At Helsinki University Central Hospital during the period 1999 to 2003, 49 patients with RA were switched from infliximab to etanercept. The reasons for infliximab discontinuation were: 42% for failure to respond by >American College of Rheumatology 50% criteria; 12% for adverse events; 46% responded to infliximab and were switched for non-medical reasons. Clinical outcome after the switch was compared between the groups according to the reason of infliximab discontinuation. Disease activity was measured with the 28-joint count Disease Activity Score (DAS28). In patients in the non-medical reasons group, the disease activity was suppressed effectively both during infliximab and etanercept. Furthermore, the one-year drug survival of etanercept in this group was the highest of 77% (95% confidence interval (CI), 62 to 97) among the three groups. In patients in the infliximab failure and adverse event groups, DAS28 values improved significantly during etanercept therapy. The 1-year drug survival of etanercept was 43% (95% CI, 26 to 70) and 50% (95% CI, 33 to 100), respectively. For RA patients who discontinued taking infliximab because of non medical reasons experienced similar treatment efficacy during both biological agents. The treatment with etanercept provided sufficient disease control also for patients with infliximab failure or adverse event. Therefore, etanercept can be suggested when infliximab has failed or discontinued for other reasons. PMID- 18414968 TI - A histomorphometric analysis of synovial biopsies from individuals with Gulf War Veterans' Illness and joint pain compared to normal and osteoarthritis synovium. AB - We compared histologic, immunohistochemical, and vascular findings in synovial biopsies from individuals with Gulf War Veterans Illness and joint pain (GWVI) to findings in normal and osteoarthritis (OA) synovium. The following parameters were assessed in synovial biopsies from ten individuals with GWVI: lining thickness, histologic synovitis score, and vascular density in hematoxylin & eosin-stained sections; and CD68+ lining surface cells and CD15+, CD3+, CD8+, CD20+, CD38+, CD68+, and Ki-67+ subintimal cells and von Willebrand Factor+ vessels immunohistochemically. Comparisons were made to synovial specimens from healthy volunteers (n = 10) and patients with OA or RA (n = 25 each). Histologic appearance and quantitative assessments were nearly identical in the GWVI and normal specimens. Vascular density was between 25% (H & E stains; p = 0.003) and 31% (vWF immunostains; p = 0.02) lower in GWVI and normal specimens than in OA. CD68+ macrophages were the most common inflammatory cells in GWVI (45.3 +/- 10.1 SEM cells/mm(2)) and normal synovium (45.6 +/- 7.4) followed by CD3+ T cells (GWVI, 15.1 +/- 6.3; normal, 27.1 +/- 9.2), whereas there were practically no CD20+, CD38+, and CD15+ cells. All parameters except lining thickness and CD15 and CD20 expression were significantly higher in OA. Five (20%) OA specimens contained significant fractions of humoral immune cells in mononuclear infiltrates, although the overall differences in the relative composition of the OA mononuclear infiltrates did not reach statistical significance compared to GWVI and normal synovium. In summary, the GWVI and normal synovia were indistinguishable from each other and contained similar low-grade inflammatory cell populations consisting almost entirely of macrophages and T cells. PMID- 18414969 TI - Deficiencies in the education of musculoskeletal medicine in Ireland. AB - BACKGROUND: Basic competency in musculoskeletal medicine is essential for many specialties being particularly relevant to primary care. AIM: The purpose of this study was to objectively assess the adequacy of musculoskeletal education at multiple levels of medical training from undergraduate level to primary care. METHODS: A previously validated musculoskeletal examination was administered to 303 volunteers consisting of medical students, orthopaedic specialist registrars, general practice trainees and general practitioners. RESULTS: Forty (71%) general practitioners and 74 (71.8%) general practice trainees failed to obtain the passing score of 70. Sixty-three (87.5%) medical students who had completed an intensive 1-week long course in musculoskeletal medicine failed the examination. The pass rate improved significantly for general practitioners who had completed a postgraduate rotation in musculoskeletal medicine (47.8 vs 18.1%, P < 0.01). CONCLUSIONS: These findings suggest that training in musculoskeletal medicine is inadequate at multiple levels of medical education with reform urgently required. PMID- 18414970 TI - Survey of medication documentation at hospital discharge: implications for patient safety and continuity of care. AB - BACKGROUND: Medication discrepancies at the time of hospital discharge are common and can result in error, patient/carer inconvenience or patient harm. Providing accurate medication information to the next care provider is necessary to prevent adverse events. AIMS: To investigate the quality and consistency of medication details generated for such transfer from an Irish teaching hospital. METHODS: This was an observational study of 139 cardiology patients admitted over a 3 month period during which a pharmacist prospectively recorded details of medication inconsistencies. RESULTS: A discrepancy in medication documentation at discharge occurred in 10.8% of medication orders, affecting 65.5% of patients. While patient harm was assessed, it was only felt necessary to contact three (2%) patients. The most common inconsistency was drug omission (20.9%). CONCLUSIONS: Inaccuracy of medication information at hospital discharge is common and compromises quality of care. PMID- 18414971 TI - Single-center 10-year experience in the management of anterior abdominal wall defects. AB - BACKGROUND: Anterior abdominal wall defects can be managed with various techniques, since no one technique can be applied to all types of defects. The aim of this study is to present a single-center experience using biomaterials and evaluate the outcomes. METHODS: The clinical data obtained from 33 neonates (24 gastroschisis and 9 omphaloceles) managed for anterior abdominal wall defects at a single center from 1995 until 2005 were evaluated. RESULTS: Primary skin closure of the defect was possible in 22 (67%) cases; however, 4 of 22 (12%) required an inner patch. In seven (21%), two patches were employed. The grafts used were: seven (22%) Goretex, four (12%) dura and one (3%) bovine pericard. CONCLUSION: In our single-center experience, both synthetic (Goretex patches) and natural biomaterials (dura and bovine patches) were found to be alternatives to manage these defects. The advantages and disadvantages of the alternatives are presented. PMID- 18414972 TI - Complete sequence analysis reveals two distinct poleroviruses infecting cucurbits in China. AB - The complete RNA genomes of a Chinese isolate of cucurbit aphid-borne yellows virus (CABYV-CHN) and a new polerovirus tentatively referred to as melon aphid borne yellows virus (MABYV) were determined. The entire genome of CABYV-CHN shared 89.0% nucleotide sequence identity with the French CABYV isolate. In contrast, nucleotide sequence identities between MABYV and CABYV and other poleroviruses were in the range of 50.7-74.2%, with amino acid sequence identities ranging from 24.8 to 82.9% for individual gene products. We propose that CABYV-CHN is a strain of CABYV and that MABYV is a member of a tentative distinct species within the genus Polerovirus. PMID- 18414973 TI - Deviation from the random distribution pattern of influenza A virus gene segments in reassortants produced under non-selective conditions. AB - High-frequency reassortment of gene segments is characteristic for influenza viruses, and it is considered to be of significance for the origin of pandemic influenza. In order to analyze whether the segregation of genes in the reassortants is random, or it deviates from the random pattern, we inoculated embryonated chicken eggs simultaneously with two influenza viruses, A/WSN/33 (H1N1) and A/Duck/ Czechoslovakia/56 (H4N6), at a high multiplicity of infection. The virus yield was used for plaque cloning, and the genetic content of plaque isolates was determined by analysis of the mobility of virus-induced proteins in polyacrylamide gel (for NP and NS genes), partial sequencing (for M gene) and polymerase chain reaction analysis with strain-specific primers for the other genes. Out of 37 isolates, 27 were reassortants. The majority of the reassortants contained the HA gene of A/WSN/33 (H1N1) virus and the NP gene of A/Duck/Czechoslovakia/56 (H4N6) virus. The data demonstrate the previously unrecognized phenomenon of segment-specific deviation from the random distribution of parent genes in the reassortants. The results are discussed in connection with the problem of differential competition between influenza A virus gene segments in mixed infection and random versus non-random reassortment of gene segments under non-selective conditions. PMID- 18414974 TI - Taurine plays a beneficial role against cadmium-induced oxidative renal dysfunction. AB - The present study has been carried out to investigate the role of taurine (2 aminoethanesulfonic acid), a conditionally essential amino acid, in ameliorating cadmium-induced renal dysfunctions in mice. Cadmium chloride (CdCl(2)) has been selected as the source of cadmium. Intraperitoneal administration of CdCl(2 )(at a dose of 4 mg/kg body weight for 3 days) caused significant accumulation of cadmium in renal tissues and lessened kidney weight to body weight ratio. Cadmium administration reduced intracellular ferric reducing/antioxidant power (FRAP) of renal tissues. Levels of serum marker enzymes related to renal damage, creatinine and urea nitrogen (UN) have been elevated due to cadmium toxicity. Cadmium exposure diminished the activities of enzymatic antioxidants, superoxide dismutase (SOD), catalase (CAT), glutathione-S-transferase (GST), glutathione reductase (GR), glutathione peroxidase (GPx) and glucose-6-phosphate dehydrogenase (G6PD) as well as non-enzymatic antioxidant, reduced glutathione (GSH) and total thiols. On the other hand, the levels of oxidized glutathione (GSSG), lipid peroxidation, protein carbonylation, DNA fragmentation, concentration of superoxide radicals and activities of cytochrome P450 enzymes (CYP P450s) have been found to increase due to cadmium intoxication. Treatment with taurine (at a dose of 100 mg/kg body weight for 5 days) before cadmium intoxication prevented the toxin-induced oxidative impairments in renal tissues. The beneficial role of taurine against cadmium-induced renal damage was supported from histological examination of renal segments. Vitamin C, a well-established antioxidant was used as the positive control in the study. Experimental evidence suggests that both taurine and vitamin C provide antioxidant defense against cadmium-induced renal oxidative injury. Combining all, results suggest that taurine protects murine kidneys against cadmium-induced oxidative impairments, probably via its antioxidative property. PMID- 18414975 TI - Comparison of the carnosine and taurine contents of vastus lateralis of elderly Korean males, with impaired glucose tolerance, and young elite Korean swimmers. AB - The carnosine and taurine contents of the vastus lateralis of two diverse groups of Korean male subjects (elderly and impaired glucose-tolerant (IGT) subjects and young elite swimmers at a national sport university) having a similar national diet, were examined. Despite marked differences in age, fitness and clinical status the two groups showed almost identical muscle carnosine and taurine contents. In the case of carnosine, the results suggest a similar contribution to intracellular buffering capacity in the two groups of subjects, with no evidence of a reduction of this in elderly IGT subjects. In addition, both groups showed the same inverse relationship between the muscle carnosine and taurine contents; the spread of values between subjects, within-groups, most likely reflect variations in the type I (low carnosine, high taurine) or type II (high carnosine, low taurine) composition of the vastus lateralis. The relationship is consistent with a role of taurine in osmoregulation, compensating for variations between fibre types in the carnosine content. PMID- 18414976 TI - Mice transgenic for reduced folate carrier: an animal model of Down syndrome? AB - In a previous publication we observed aberrant levels of the human reduced folate carrier (hRFC) in cortex from fetal Down syndrome (DS) subjects. Immunoreactivity for hRFC was increased as the only chromosome 21 gene product studied. We, therefore, analyzed mice transgenic for hRFC (TghRFC1) and wild-type (WT) mice for cognitive functions, behavior and in an observational neurological battery (FOB). Cognitive functions were evaluated by the Morris water maze (MWM), the open field (OF) was used for exploratory behavior, locomotor activity and anxiety related behavior. The elevated plus maze (EPM) was used to confirm findings in the OF testing anxiety-related behavior and the rota rod (RR) to evaluate motor function. In the MWM TghRFC1 mice performed significantly worse (P < 0.0003) on the probe trial than WT mice. In the FOB visual placing was significantly reduced inTghRFC1 mice. In the OF TghRFC1 mice crossed twice as often (P < 0.029) and in the EPM individuals from this group showed a reduced number of exits from the closed arm (P < 0.044) compared to WT mice. TghRFC1 mice showed impaired performance on the RR, spending one-fourth of the time of WT on the revolving rod (P < 0.0003). Cognitive impairment is an obligatory symptom of DS and this deficiency corresponds to findings in the MWM of mice transgenic for hRFC. Findings of visual placing and failure on the RR may reflect impaired motor performance including muscular hypotonia in DS subjects. Increased crossings in the OF may indicate modulated anxiety-related behavior observed in patients with DS. PMID- 18414977 TI - Process improvement in amino acid N-carboxyanhydride synthesis by N-carbamoyl amino acid nitrosation. AB - Amino acid N-carboxyanhydrides (NCA), convenient monomer for polypeptide synthesis, are easily prepared in high purity as the result of N-carbamoyl amino acids (CAA) nitrosation by gaseous NOx (4:1 NO + O2 mixture, or NOCl) in toluene. Removal of polar side products is then efficiently carried out during subsequent work-up and crystallisation so that the resulting NCA obtained in good yield is suitable for controlled, primary amine-initiated polymerisation. PMID- 18414978 TI - Systematic comparison of two novel, thiol-reactive prosthetic groups for 18F labeling of peptides and proteins with the acylation agent succinimidyl-4 [18F]fluorobenzoate ([18F]SFB). AB - A systematic comparison of 4-[18F]fluorobenzaldehyde-O-(2-{2-[2-(pyrrol-2,5-dione 1-yl)ethoxy]-ethoxy}-ethyl)oxime ([18F]FBOM) and 4-[18F]fluorobenzaldehyde-O-[6 (2,5-dioxo-2,5-dihydro-pyrrol-1-yl)-hexyl]oxime ([18F]FBAM) as prosthetic groups for the mild and efficient 18F labeling of cysteine-containing peptides and proteins with the amine-group reactive acylation agent, succinimidyl-4 [18F]fluorobenzoate ([18F]SFB), is described. All three prosthetic groups were prepared in a remotely controlled synthesis module. Synthesis of [18F]FBOM and [18F]FBAM was accomplished via oxime formation through reaction of appropriate aminooxy-functionalized labeling precursors with 4-[18F]fluorobenzaldehyde. The obtained radiochemical yields were 19% ([18F]FBOM) and 29% ([18F]FBAM), respectively. Radiolabeling involving [18F]FBAM and [18F]FBOM was exemplified by the reaction with cysteine-containing tripeptide glutathione (GSH), a cysteine containing dimeric neurotensin derivative, and human native low-density lipoprotein (nLDL) as model compounds. Radiolabeling with the acylation agent [18F]SFB was carried out using a dimeric neurotensin derivative and nLDL. Both thiol-group reactive prosthetic groups show significantly better labeling efficiencies for the peptides in comparison with the acylation agent [18F]SFB. The obtained results demonstrate that [18F]FBOM is especially suited for the labeling of hydrophilic cysteine-containing peptides, whereas [18F]FBAM shows superior labeling performance for higher molecular weight compounds as exemplified for nLDL apolipoprotein constituents. However, the acylation agent [18F]SFB is the preferred prosthetic group for labeling nLDL under physiological conditions. PMID- 18414979 TI - Flavonols from Bauhinia malabarica. AB - Bauhinia malabarica Roxb. has been used in Thai traditional medicine for wound healing, as a diuretic, to fight dysentery and as an emmenagogue. Seven flavonols, including 6,8-di-C-methylkaempferol 3-methyl ether, kaempferol, afzelin, quercetin, isoquercitrin, quercitrin, and hyperoside were isolated from the methanol extract of leaves. PMID- 18414980 TI - Primary non-Hodgkin's lymphoma of the breast: eight-year follow-up experience. AB - The objective of this study is to analyze the clinical characteristics and treatment of patients with primary non-Hodgkin's lymphoma of the breast (PNHLB). Forty-five patients with PNHLB treated in our hospital during a 15-year period were retrospectively analyzed. Forty-four were females and one male, with a median age of 47 years. Forty-two patients were at stage I or II and 82.2% had diffuse large B cell lymphoma (DLBCL). Local control rate was 95.2 and 66.7% for patients with and without radiotherapy, respectively (P = 0.020). Median overall survival and progression-free survival (PFS) of all patients was 6.8 and 4.3 years, respectively. For patients with DLBCL or T cell lymphoma, median PFS was 6.5 years with chemoradiation and 3.9 years with chemotherapy or radiation only (P = 0.029). Patients who used rituximab had not reached median PFS, while those treated without rituximab had a median PFS of 5.1 years (P = 0.301). International prognostic index (IPI) score and bilateral breast involvement were two independent prognostic factors for survival. Chinese patients with PNHLB have early occurrence in lifespan. Radiation confers a better local control. Patients with intermediate or high-grade PNHLB might be treated with chemotherapy, radiotherapy, and for CD-20 positive disease, rituximab. Bilateral disease and IPI are two prognostic factors. PMID- 18414981 TI - Successful response to rituximab in two cases of acquired haemophilia refractory to standard-therapy. AB - Acquired autoantibodies against coagulation factors (acquired haemophilia) frequently constitute a life-threatening bleeding situation requiring a prompt therapeutic intervention, including control of bleeding and secondarily an attempt of eradication of the inhibitor by prolonged immunosuppressive therapy. The combination of oral corticosteroids and cyplophosphamide seems to be effective to eradicate the autoantibody, but some patients may be resistant. Another therapeutic approach, recently described, observes treatment with the chimeric anti-CD20 monoclonal antibody rituximab. We report two consecutively treated patients whose acquired FVIII inhibitors did not respond to standard immunosuppressive regimens, and only when rituximab was added to therapy, complete response and prolonged remission were obtained. PMID- 18414982 TI - Aberrant expression of BCL2A1-restricted minor histocompatibility antigens in melanoma cells: application for allogeneic transplantation. AB - It has been shown that allogeneic hematopoietic stem cell transplantation (HSCT) can be one of the therapeutic options for patients with metastatic solid tumors, such as renal cancer. However, the development of relatively severe GVHD seems to be necessary to achieve tumor regression in the current setting. Thus, it is crucial to identify minor histocompatibility antigens (mHags) only expressed in tumor cells but not GVHD target organs. In this study, we examined whether three mHags: ACC-1 and ACC-2 encoded by BCL2A1, and HA-1 encoded by HMHA1, could serve as such targets for melanoma. Real-time PCR and immunohistochemical analysis revealed that the expression of both BCL2A1and HMHA1 in melanoma cell lines and primary melanoma cells was comparable to that of hematopoietic cells. Indeed, melanoma cell lines were efficiently lysed by cytotoxic T lymphocytes specific for ACC-1, ACC-2, and HA-1. Our data suggest that targeting mHags encoded not only by HMHA1, whose aberrant expression in solid tumors has been reported, but also BCL2A1 may bring about beneficial selective graft-versus-tumor effects in a population of melanoma patients for whom these mHags are applicable. PMID- 18414983 TI - Enhancement of anti-lymphoma immuno-effects mediated by dendritic cells pulsed with heat-stressed and rituximab-coated CD20+ lymphoma cells. AB - Rituximab, a CD20-reactive chimeric monoclonal antibody (mAb), induces apoptosis of lymphoma cells and promotes phagocytosis by dendritic cells and produces a cellular immunoresponse by cross-presentation of tumor antigens to T cells. Heat stressed tumor cells also stimulate dendritic-cell maturation and induce specific cytotoxic T cells against tumor cells by inducing expression of heat-shock proteins. In this study, we used heat-stressed and rituximab-coated CD20+ lymphoma cells as antigens to load onto immature dendritic cells, and found that rituximab-coated CD20+ Raji cells could promote phagocytosis by dendritic cells, and that rituximab-coated or heat-stressed and then rituximab-coated CD20+ Raji cells resulted in increased dendritic cell maturation. Moreover, only CD20+ Raji cells treated with heat and rituximab effectively enhanced the immunostimulating functions of dendritic cells. Thus, our data indicate that rituximab and heat shock proteins have synergistic effects, resulting in increased induction of cytotoxic T cells against B-cell lymphoma. PMID- 18414984 TI - Pulmonary embolism in a patient with multiple myeloma receiving thalidomide dexamethasone therapy. AB - Massive pulmonary embolism is an uncommon complication of multiple myeloma treated with thalidomide-dexamethasone regimen. In 2006, multiple myeloma was diagnosed in a 72-year-old man, who received thalidomide-dexamethasone therapy. In January 2007, echocardiography and computerized tomography identified massive pulmonary embolism in the pulmonary arteries and a deep vein thrombus of the right leg. The patient also had an elevated concentration of B-type natriuretic peptide. After heparinization and warfarin therapy, the patient's condition improved. This is the first report of a patient with a rare complication of pulmonary embolism from thalidomide-treated multiple myeloma. PMID- 18414985 TI - In remembrance of Dr. Tetsuzo Akutsu: a man who started artificial heart research. PMID- 18414986 TI - Journal of Artificial Organs 2007: the year in review. PMID- 18414987 TI - Percutaneous and off-pump treatments for functional mitral regurgitation. AB - A new era in the treatment of functional mitral regurgitation is emerging with new devices that can be placed percutaneously or minimally invasively without cardiopulmonary bypass. These devices are categorized into three groups: annuloplasty, edge-to-edge repair, and ventricular reshaping. Percutaneous annuloplasty devices, implanted via the coronary sinus, mimic surgical annuloplasty by reducing the mitral annular anterior-posterior (or septal lateral) dimension. Several devices, such as the PTMA, CARILLON, Monarch, and PS3 systems, are in clinical trials. Percutaneous edge-to-edge repair devices mimic the surgical Alfieri edge-to-edge repair technique, creating a double-orifice mitral valve; of these, MitraClip is in clinical trials. Ventricular reshaping devices treat both mitral annular dilatation and papillary muscle displacement (and thus leaflet tethering). The surgical Coapsys device is currently in clinical trials, and its percutaneous "interventional" version, iCoapsys, is being prepared for a clinical trial. Numerous issues need to be addressed before these devices can become standard therapies for functional mitral regurgitation. Device safety and efficacy must be demonstrated in carefully designed clinical trials with the goal of achieving outcomes equal to or better than those of surgical repair. PMID- 18414988 TI - Surgery for acute aortic dissection using gelatin-resorcin-formalin glue: perspective from 10 years of follow-up at a single center. AB - Acute type A aortic dissection is a lethal aortic disease with an extremely poor prognosis unless surgical intervention is performed in a timely manner. Since the clinical introduction of gelatin-resorcin-formalin (GRF) glue in Japan, tear oriented aortic repairs have been widely performed with the aid of this glue at many institutions over the past decade. A total of 138 patients underwent glue aided emergency operations for acute type A aortic dissection at Nihon University Itabashi hospital. The hospital mortality rate was 6.7%. The actuarial survival rate was 81.5% after 5 years and 54.8% after 10 years. The rate of freedom from reoperation was 87.9% after 5 years and 72.3% after 10 years. Although application of GRF glue remains controversial because of concerns about the cytotoxicity of formaldehyde and tissue necrosis, cytotoxicity and tissue necrosis have not been found at reoperation. GRF glue demonstrated excellent tissue adhesion and hemostasis capabilities and contributed to improved surgical results. Although some disadvantages have been reported, complications can be avoided by proper use of the glue and proper selection of surgical procedure, based on the pathology of the patient. PMID- 18414989 TI - Oxygen metabolism during cardiopulmonary bypass with hemodilution using liposome encapsulated hemoglobin in kid goats. AB - Cardiopulmonary bypass (CPB) with hemodilution has been proposed as a useful method for many types of cardiovascular surgery. Although the harmful effects of severe hemodilution need to be prevented, blood transfusion should be avoided whenever possible. Therefore, we have been developing a new CPB technique using liposome-encapsulated hemoglobin (LEH). The purpose of this study was to evaluate the combined therapy of diluted CPB and LEH focusing on the influence of LEH on oxygen metabolism. Male kid goats (n = 8) were divided into two groups: the LEH and control groups. CPB was maintained at between 36 degrees and 37 degrees C. There was no significant difference in hemoglobin concentrations (6.3 +/- 1.5 g/dl in the LEH group and 6.2 +/- 1.3 g/dl in the control group) after initiation of CPB between the two groups. Thus, there was no distinction in oxygen deliveries between the two groups (11.0 +/- 2.0 ml/kg/min in the LEH group and 11.0 +/- 2.3 ml/kg/min in the control group). Oxygen consumption in the LEH group (2.5-2.7 ml/kg/min), however, had a tendency to be higher than that in the control group (2.4-2.5 ml/kg/min). In addition, the lactate/pyruvate ratio decreased earlier in the LEH group. These results suggest that the application of LEH in the pump-priming solution improves decreased aerobic oxygen metabolism during CPB without any serious adverse effects. PMID- 18414990 TI - Wavelet analysis of valve closing sound detects malfunction of bileaflet mechanical valve. AB - Several studies have reported the asynchronous closure of normal bileaflet valves (NBVs), resulting in a split in its closing sound; however, the clinical significance of this split has never been studied in malfunctioning bileaflet valves (MBVs). The study comprised 218 valves in 184 patients, including normal monoleaflet valves (n = 10), NBVs (n = 198), and MBVs (n = 10). Valve function was confirmed by cinefluoroscopy prior to analysis of the valve sound by the Morlet continuous wavelet transform (CWT). The split interval (SI) for each heartbeat was measured, and the coefficient of variation (CV) of its mean (valve SI) was calculated as a parameter for the fluctuation of the SI. The CWT of monoleaflet valves showed a single spike, whereas NBVs exhibited a clear split. There was no significant difference in valve SI between the aortic and mitral positions; however, the mean of the CV was significantly greater in the mitral position (n = 90, 0.507 +/- 0.254) than in aortic position (n = 108, 0.353 +/- 0.228, P = 0.000045). The split was not found in six (aortic; three, mitral; three) of ten patients with MBVs. The other four patients had a distinct split, but the CV was significantly lower for MBVs (0.138 +/- 0.105) than for NBVs (0.343 +/- 0.221, P = 0.042). Receiver-operating characteristics analysis demonstrated the cutoff line of the CV to be 0.112 for detecting malfunctioning aortic valves with the highest accuracy of 86.1%. This new system using the Morlet CWT can detect MBVs. It will be a useful modality for screening the function of bileaflet valves. PMID- 18414991 TI - Glial reaction to photoelectric dye-based retinal prostheses implanted in the subretinal space of rats. AB - We have designed a new type of retinal prosthesis using polyethylene films coupled with photoelectric dye molecules that absorb light and convert photon energy to electric potentials. An extruded-blown film of high-density polyethylene was used as the original polyethylene film. Recrystallized film was made by recrystallization from the melting of the original polyethylene film. A photoelectric dye,2-[2-[4-(dibutylamino)phenyl]ethenyl]-3 carboxymethylbenzothiazolium bromide, was coupled to the two types of polyethylene films through amide linkages. Samples of the original dye-coupled film, the dye-coupled recrystallized film, and the dye-uncoupled plain film were implanted in the subretinal space of normal adult rats. Frozen sections were cut from the eyes enucleated at 1 week or 1 month and were either stained with hematoxylin and eosin, stained immunohistochemically for glial fibrillary acidic protein (GFAP), or processed for in situ apoptosis detection. The results revealed that retinal tissue damage was negligible with no inflammatory cells and few apoptotic cells. GFAP was significantly up-regulated in retinal sites with the implantation of all types of polyethylene films at 1 week, compared with the adjacent retinal sites (P < 0.005, analysis of variance). The GFAP up-regulation was also present at 1 month for the plain film and dye-coupled recrystallized film (P < 0.05). Glial cell encirclement around the films increased significantly between 1 week and 1 month (P = 0.023, two-factor analysis of variance) but was not significantly different among the three types of polyethylene films (P = 0.4531). These results showed evidence of glial reactions to the photoelectric dye-coupled polyethylene films implanted into the subretinal space of rat eyes and also proved their basic biological safety. PMID- 18414992 TI - Short-term biological safety of a photoelectric dye used as a component of retinal prostheses. AB - We have designed a new type of retinal prosthesis with a photoelectric dye that transfers photon energy to generate electric potentials. The purpose of this study was to test the safety of a photoelectric dye, 2-[2-[4-(dibutylami no)phenyl]ethenyl]-3-carboxymethylbenzothiazolium bromide (NK-5962), used for retinal prostheses. The retinal cells, derived from chick neurosensory retinas at the 12-day embryonic stage, were a mixed population of retinal neurons and glial cells, and were cultured for 2 days either under protection from light or under continuous light exposure at 230 lux for 9 h daily in the presence of the photoelectric dye at varying concentrations (1.6 x 10(-5), 1.6 x 10(-6), and 1.6 x 10(-7) M) to assess cell viability by staining live cells and dead cells. Dispersed retinal pigment epithelial cells at the same embryonic stage were incubated with the photoelectric dye at varying concentrations (6.6 x 10(-5), 6.6 x 10(-6), and 6.6 x 10(-7) M) for 4 h under protection from light or under continuous light exposure at 320 lux to assess cytotoxicity by measuring the activity of lactate dehydrogenase leaking from cells. The majority of retinal cells were alive with only a small percentage of dead cells under the dark condition or the light condition in the presence or the absence of the photoelectric dye. The percentage of dead cells was significantly smaller at higher concentrations of the photoelectric dye (P = 0.0183, two-factor analysis of variance), while the percentage of dead cells was not significantly different between the dark condition and the light condition (P = 0.3102). Percent cytotoxicity values were negative, indicating protective effects in all groups of retinal pigment epithelial cells incubated with varying concentrations of the photoelectric dye. The photoelectric dye showed no cytotoxicity to chick retinal cells or retinal pigment epithelial cells on short-term exposure. In addition, this photoelectric dye might have protective effects on both types of cells. PMID- 18414994 TI - A decrease in S-adenosyl-L-methionine potentiates arachidonic acid cytotoxicity in primary rat hepatocytes enriched in CYP2E1. AB - Previous studies show that treatment with a polyunsaturated fatty acid, arachidonic acid (AA), or high concentrations of cycloleucine, an inhibitor of methionine adenosyltransferase (MAT), which lowers levels of S-adenosyl-L methionine (SAM), increased toxicity in hepatocytes from pyrazole-treated rats which expressed high levels of cytochrome P450 2E1 (CYP2E1). In this study, I used concentrations of cycloleucine or AA, which by themselves do not produce any toxicity, to evaluate whether a decrease in SAM sensitizes hepatocytes to AA toxicity, especially in hepatocytes enriched in CYP2E1. Levels of SAM were lower by 50% in hepatocytes from pyrazole- compared to saline-treated rats. Cycloleucine treatment caused a 50% decline in SAM levels with both hepatocyte preparations and SAM levels were lowest in the pyrazole-treated hepatocytes. The combination of cycloleucine plus AA produced some toxicity and apoptosis in hepatocytes from saline-treated rats but increased toxicity and apoptosis was found in the hepatocytes from pyrazole-treated rats. Cytotoxicity could be prevented by incubation with SAM, the antioxidant trolox, and the mitochondrial permeability transition inhibitor trifluoperazine. The enhanced cytotoxicity could also be protected by treating rats with chlormethiazole, a specific inhibitor of CYP2E1, thus validating the role of CYP2E1. Cycloleucine plus AA treatment elevated production of reactive oxygen species (ROS) and lipid peroxidation to greater extents with the hepatocytes from pyrazole-treated rats than that from the saline-treated rats. I hypothesize that increased production of ROS by hepatocytes enriched in CYP2E1 potentiates AA-induced lipid peroxidation and toxicity when hepatoprotective levels of SAM are lowered. Such interactions, e.g. induction of CYP2E1, decline in SAM and polyunsaturated fatty acid-induced lipid peroxidation, may contribute to alcohol-induced liver injury. PMID- 18414995 TI - Rafts making and rafts braking: how plant flavonoids may control membrane heterogeneity. AB - Plant flavonoids are not only known as powerful antioxidants, but also as cell metabolism regulators. It has been postulated that they are able to control cell signal pathways by targeting receptors on the cell surface or by intercalating the lipid bilayer of membranes. Some flavonoids can increase lipid viscosity and decrease the cooperativity of hydrocarbon chain melting, while others can considerably decrease the lipid melting temperature, thus providing additional freedom for lipid diffusion. Here we discuss the ability of flavonoids to influence phase transition and lateral segregation of lipids, responsible for the formation of membrane compartments known as lipid rafts. The thermodynamic parameters of the bilayer determined by lipid packing characteristics and by lateral segregation of the bilayer are expected to depend on the location of flavonoid molecules in the bilayer. Flavonoid molecules preferably located in the hydrophobic region of the bilayer can initiate formation of raft-like domains (raft-making effect), while the molecules located in the polar interface region of the bilayer can fluidize membranes (raft-breaking effect), or initiate formation of interdigitated or micellar structures. Accordingly, we expect that in cellular membranes flavonoids can influence the appearance and development of rafts or raft-like membrane domains and thus influence the lateral diffusion of lipid molecules. Because rafts participate in cellular signal transduction, endocytosis and transmembrane translocation of different compounds, flavonoids may control cell metabolism by modulating the bilayer state. PMID- 18414996 TI - Decylubiquinol impedes mitochondrial respiratory chain complex I activity. AB - We have studied the interaction of decylubiquinone, an effective substrate for respiratory chain complexes III and II, with complex I in mouse and human tissues. We found that its reduced form, decylubiquinol, severely impedes complex I activity, while the oxidized form, decylubiquinone acts as a potent acceptor for complex I electrons. This observation has obvious incidence on the assay conditions for complex I. In keeping with that, we found that the inhibition by the reduced form can be avoided by maintaining decylubiquinone under an oxidized form. Under these experimental conditions, a high complex I activity could be measured allowing to detect partial complex I deficiency. Use of these conditions is however restricted to tissues/cells with limited contaminating NADH dehydrogenase activities that are prone to react with redox active compounds. PMID- 18414993 TI - Clinical biomarkers of angiogenesis inhibition. AB - INTRODUCTION: An expanding understanding of the importance of angiogenesis in oncology and the development of numerous angiogenesis inhibitors are driving the search for biomarkers of angiogenesis. We review currently available candidate biomarkers and surrogate markers of anti-angiogenic agent effect. DISCUSSION: A number of invasive, minimally invasive, and non-invasive tools are described with their potential benefits and limitations. Diverse markers can evaluate tumor tissue or biological fluids, or specialized imaging modalities. CONCLUSIONS: The inclusion of these markers into clinical trials may provide insight into appropriate dosing for desired biological effects, appropriate timing of additional therapy, prediction of individual response to an agent, insight into the interaction of chemotherapy and radiation following exposure to these agents, and perhaps most importantly, a better understanding of the complex nature of angiogenesis in human tumors. While many markers have potential for clinical use, it is not yet clear which marker or combination of markers will prove most useful. PMID- 18414997 TI - Anaplerosis from glucose, alpha-ketoisocaproate, and pyruvate in pancreatic islets, INS-1 cells and liver mitochondria. AB - Methyl succinate (MS) and alpha-ketoisocaproate (KIC) when applied alone to cultured pancreatic islets or INS-1 832/13 cells do not stimulate insulin release. However, when the two metabolites are combined together they strongly stimulate insulin release. Studying the possible explanations for this complementarity has provided clues to the pathways involved in insulin secretion. MS increased carbon incorporation of KIC into acid-precipitable material and lipid in INS-1 cells. In isolated mitochondria, MS alone increased malate, but MS plus KIC increased citrate, alpha-ketoglutarate, and isocitrate. These data and the known pathways of their metabolism suggest that MS supplies the oxaloacetate component of citrate and KIC supplies the acetate component of citrate. Other citric acid cycle intermediates can be formed from citrate enabling anaplerosis to supply precursors for extramitochondrial pathways. In addition, KIC, glucose and pyruvate can be metabolized to acetoacetate. In an INS-1 cell line deficient in ATP citrate lyase, incorporation of carbon from pyruvate into acid precipitable material and lipid was not lowered. This negative result is in agreement with our recent discovery that citrate is not the only carrier of acyl groups from the mitochondria to the cytosol in the beta cell and that acetoacetate can also transfer acyl carbon to the cytosol. PMID- 18414998 TI - Epstein-Barr virus induces an oxidative stress during the early stages of infection in B lymphocytes, epithelial, and lymphoblastoid cell lines. AB - The study investigates the direct effect of Epstein-Barr virus infection on the oxidative profile of in vitro cultivated human cells. For this purpose, a panel of human EBV target cells presenting heterogeneity in their cellular and culture types (epithelial cells or lymphocytes; primary culture or continuous cell culture) was selected. These cells are purified human B lymphocytes, DG75, 293, and HepG2 cell lines. The oxidative stress was evaluated during the early stages of infection (2, 12, and 24 h) by measuring malondialdehyde, the end product of the lipid peroxidation, as well as the activities of two antioxidant enzymes: catalase and superoxide dismutase. The obtained results were compared with those of the untreated cells and the K562 cell line which has no interaction with EBV. The incubation of the different target cells with EBV induced an oxidative stress in the purified B lymphocytes, DG75, and 293, but not in HepG2 and K562. This oxidative stress was highlighted by an increase in MDA level (P < 0.05), which began 2 h after the addition of the virus and persisted after 12 and 24 h. Simultaneously, a decrease in catalase and superoxide dismutase activities was observed (P < 0.05), suggesting an alteration of the molecular mechanisms promoting cellular resistance to reactive oxygen species (ROS). The efficiency of EBV infection, assessed by viral DNA PCR amplification, was confirmed in 293 and DG75 but not in HepG2, which was in total concordance with their oxidative profiles. In conclusion, the EBV infection of B and epithelial cells leads to the establishment of an oxidative stress which can play a key role during the viral transformation. PMID- 18414999 TI - Influence of nanoporesize on platelet adhesion and activation. AB - In this study we have evaluated the influence of biomaterial nano-topography on platelet adhesion and activation. Nano-porous alumina membranes with pore diameters of 20 and 200 nm were incubated with whole blood and platelet rich plasma. Platelet number, adhesion and activation were determined by using a coulter hematology analyzer, scanning electron microscopy, immunocytochemical staining in combination with light microscopy and by enzyme immunoassay. Special attention was paid to cell morphology, microparticle generation, P-selectin expression and beta-TG production. Very few platelets were found on the 200 nm alumina as compared to the 20 nm membrane. The platelets found on the 20 nm membrane showed signs of activation such as spread morphology and protruding filipodia as well as P-selectin expression. However no microparticles were detected on this surface. Despite the fact that very few platelets were found on the 200 nm alumina in contrast to the 20 nm membrane many microparticles were detected on this surface. Interestingly, all microparticles were found inside circular shaped areas of approximately 3 mum in diameter. Since this is the approximate size of a platelet we speculate that this is evidence of transient, non-adherent platelet contact with the surface, which has triggered platelet microparticle generation. To the authors knowledge, this is the first study that demonstrates how nanotexture can influence platelet microparticle generation. The study highlights the importance of understanding molecular and cellular events on nano-level when designing new biomaterials. PMID- 18415000 TI - Osteoblast behaviour on in situ photopolymerizable three-dimensional scaffolds based on D,L-lactide and epsilon-caprolactone: influence of pore volume, pore size and pore shape. AB - Bone marrow cells were cultured on in situ photopolymerizable scaffolds based on D,L-lactide and epsilon-caprolactone. The influence of pore volume, size and shape were evaluated. Bone formation was demonstrated by ALP activity, osteocalcin secretion and histological analysis. TEM at the polymer interface revealed osteoblasts which secreted an extracellular matrix containing matrix vesicles loaded with apatite. Cellular infiltration was possible for scaffolds with a porosity of 70 and gelatin particle size of 250-355 microm. Scaffolds with a porosity less than 70 had the tendency to form a polymer top layer. Although increasing the gelatin particle size to 355-500 microm, leads to infiltration even in scaffolds with a porosity of 60. No infiltration was possible in scaffolds with sodium chloride as porogen. On the contrary, sucrose and gelatin leads to better interconnected scaffolds at the same porosity. Hence, spherical gelatin particles are suitable to use as porogen in photopolymerizable scaffolds. PMID- 18415001 TI - In vitro dissolution behavior of SiO2-MgO-Al2O 3-K2O-B2O3-F glass-ceramic system. AB - Herein, we report the results of the in vitro dissolution tests, which were carried out by immersing the selected glass-ceramic samples in artificial saliva (AS) for various time periods of up to 42 days. In our experiments, the SiO(2) MgO-Al(2)O(3)-K(2)O-B(2)O(3)-F glass ceramics with different crystal morphology and crystal content were used and a comparison is also made with the baseline glass samples (without any crystals). The bioactivity of the samples was probed by measuring the changes in pH, ionic conductivity and ionic concentration of AS following in vitro dissolution experiments. High resistance of the selected glass ceramic samples against in vitro leaching has been demonstrated by minimal weight loss (<1%) and insignificant density change, even after 6 weeks of dissolution in artificial saliva. While XRD analysis reveals the change in surface texture of the crystalline phase, FT-IR analysis weakly indicated the Ca-P compound formation on the leached surface. The experimental measurements further indicate that the leaching of F(-), Mg(2+) ions from the sample surface commonly causes the change in the surface chemistry. Furthermore, the presence of (Ca, P, O)-rich mineralized deposits on the leached glass-ceramic surface as well as the decrease in Ca(2+) ion concentrations in the leaching solutions (compared to that in the initial AS solution) provide evidences of the moderate bioactive or mild biomineralisation behaviour of investigated glass-ceramics. PMID- 18415002 TI - On the possibility of silicon nitride as a ceramic for structural orthopaedic implants. Part II: chemical stability and wear resistance in body environment. AB - In Part I, the processing, microstructure and mechanical properties of three silicon nitride-based ceramics were examined and their non-toxicity was demonstrated. In this Part II, some features critical to biomedical applications were investigated: (i) the wetting behaviour against aqueous media, including physiological solutions; (ii) the chemical stability in water and in physiological solutions; and (iii) the wear resistance, measured under experimental procedures that simulate the conditions typical of the hip joint prosthesis. The results confirmed that silicon nitride may serve as a biomaterial for bone substitution in load bearing prosthesis. PMID- 18415003 TI - Towards a controlled photopolymerization of dental dimethacrylate monomers: EPR studies on effects of dilution, filler loading, storage and aging. AB - Electron paramagnetic resonance (EPR) was used to study the kinetics of methacrylate radical formation in the monomer mixture 2,2-bis [4-(2-hydroxy-3 methacryloxyprop-1-oxy) phenyl] propane (Bis-GMA)/triethylene glycol dimethacrylate (TEGDMA), in the presence of a photo-initiator system (camphorquinone, CQ/N,N-dimethyl-p-toluidine, DET). Curing-time dependences on the filler (0-40 wt%) and TEGDMA content (15-90 wt%) were evaluated; the influence of irradiation protocol, uncured sample storage time and aging of cured systems were also studied. EPR enabled observing at least two different kinetic regimes during polymerization. The final radical concentration decreased both with Bis-GMA and filler content. However, a reverse trend was obtained when the relative photo-initiator concentrations were considered. Filler also showed a significant effect on the radical life-time reduction. Irradiation protocol and storage time of uncured matrices showed to affect the free radical concentration. The observed changes on the EPR signal lineshape with post-curing time suggests that the distribution of CH(2) conformations also changes with time. PMID- 18415004 TI - Bioactive borate glass coatings for titanium alloys. AB - Bioactive borate glass coatings have been developed for titanium and titanium alloys. Glasses from the Na(2)O-CaO-B(2)O(3) system, modified by additions of SiO(2), Al(2)O(3), and P(2)O(5), were characterized and compositions with thermal expansion matches to titanium were identified. Infrared and X-ray diffraction analyses indicate that a hydroxyapatite surface layer forms on the borate glasses after exposure to a simulated body fluid for 2 weeks at 37 degrees C; similar layers form on 45S5 Bioglass((R)) exposed to the same conditions. Assays with MC3T3-E1 pre-osteoblastic cells show the borate glasses exhibit in vitro biocompatibility similar to that of the 45S5 Bioglass((R)). An enameling technique was developed to form adherent borate glass coatings on Ti6Al4V alloy, with adhesive strengths of 36 +/- 2 MPa on polished substrates. The results show these new borate glasses to be promising candidates for forming bioactive coatings on titanium substrates. PMID- 18415007 TI - The neuregulin family of genes and their multiple splice variants in breast cancer. AB - The neuregulin family consists of four genes, NRG1-4 which can each encode products containing a domain related to the epidermal growth factor family of ligands. Each gene is subject to complex control of transcription and to splicing of their mRNA product to give many variant proteins. These do not contain secretory sequences but some, through their transmembrane sequence, are routed via the Golgi where they are glycosylated, to the cell surface. Here they may be released by regulated proteolysis to act as soluble proteins which can interact and activate members of the EGF receptor family of receptor tyrosine kinases. Other splice variants do not encode transmembrane sequences and these are found either in the cytoplasm or, if they encode a nuclear localisation sequence, in distinct compartments in the nucleoplasm. It has been shown that the variants containing a full EGF domain can act as receptor agonists but the function of the cytoplasmic and nuclear products is unknown as yet. All four neuregulin genes are expressed and play an important role in mammary gland development. They are also expressed at elevated levels in some cases of ductal carcinoma in situ of the breast and breast cancer. They seem to be active in this setting and their presence may affect the efficacy of treatment with endocrine agents or with signal transduction inhibitors directed at the EGF receptor family members. Much remains to be learned however of their normal function and their influence on breast cancer development, progression and response to therapy. PMID- 18415008 TI - The role of specific beta-gamma subunit interactions in oxyanion stimulation of the MgATP hydrolysis of a hybrid photosynthetic F1-ATPase. AB - Pairs of cysteine residues were introduced into the twisted N- and C-terminal helices of the gamma subunit of the chloroplast F1-ATPase to test, via disulfide cross-linking, potential inter-helical movements involved in catalysis of ATP hydrolysis. The extent of disulfide cross-linking was determined by estimating the amount of free sulfhydryl available for labeling with fluoresceinyl maleimide before and after cross-linking. Significant disulfide formation (50-75%) was observed between cysteines introduced at positions 30 and 31 in the N-terminal helix and 276 and 278 in the C-terminal helix. Cross-linking had no apparent effect on catalysis, therefore eliminating the involvement of large-scale inter helical movements within this region of the gamma subunit in cooperative ATP hydrolysis. However, the presence of the two cysteines together in the gammaV31C/A276C double mutant, irrespective of whether or not they were cross linked together, lowered the MgATPase activity by more than 70% and completely eliminated the well-known activating effect of the oxyanion sulfite. The CaATPase activity was unaffected. Similar but less pronounced effects were seen with the gammaK30C/A276C double mutant. The results indicate that residues at or near positions 31 and 276 within the twisted helical pair of the gamma subunit are required to overcome Mg2+ inhibition of ATP hydrolysis. These residues are likely to be involved in forming a point of contact between the gamma and beta subunits that is responsible for this effect. PMID- 18415009 TI - The nature of the memory trace and its neurocomputational implications. AB - The brain processes underlying cognitive tasks must be very robust. Disruptions such as the destruction of large numbers of neurons, or the impact of alcohol and lack of sleep do not have negative effects except when they occur in an extreme form. This robustness implies that the parameters determining the functioning of networks of individual neurons must have large ranges or there must exist stabilizing mechanisms that keep the functioning of a network within narrow bounds. The simulation of a minimal neuronal architecture necessary to study cognitive tasks is described, which consists of a loop of three cell-assemblies. A crucial factor in this architecture is the critical threshold of a cell assembly. When activated at a level above the critical threshold, the activation in a cell-assembly is subject to autonomous growth, which leads to an oscillation in the loop. When activated below the critical threshold, excitation gradually extinguishes. In order to circumvent the large parameter space of spiking neurons, a rate-dependent model of neuronal firing was chosen. The resulting parameter space of 12 parameters was explored by means of a genetic algorithm. The ranges of the parameters for which the architecture produced the required oscillations and extinctions, turned out to be relatively narrow. These ranges remained narrow when a stabilizing mechanism, controlling the total amount of activation, was introduced. The architecture thus shows chaotic behaviour. Given the overall stability of the operation of the brain, it can be concluded that there must exist other mechanisms that make the network robust. Three candidate mechanisms are discussed: synaptic scaling, synaptic homeostasis, and the synchronization of neural spikes. PMID- 18415010 TI - Gender in voice perception in autism. AB - Deficits in the perception of social stimuli may contribute to the characteristic impairments in social interaction in high functioning autism (HFA). Although the cortical processing of voice is abnormal in HFA, it is unclear whether this gives rise to impairments in the perception of voice gender. About 20 children with HFA and 20 matched controls were presented with voice fragments that were parametrically morphed in gender. No differences were found in the perception of gender between the two groups of participants, but response times differed significantly. The results suggest that the perception of voice gender is not impaired in HFA, which is consistent with behavioral findings of an unimpaired voice-based identification of age and identity by individuals with autism. The differences in response times suggest that individuals with HFA use different perceptual approaches from those used by typically developing individuals. PMID- 18415011 TI - Sexual coercion in intimate relationships: a comparative analysis of the effects of women's infidelity and men's dominance and control. AB - Researchers studying the proximate (or immediate) causes of sexual coercion have proposed that partner rape is motivated by a man's attempt to dominate and control his partner and that this expression of power is the product of men's social roles. Researchers studying the ultimate (or evolutionary) causes, in contrast, have proposed that partner rape may function as an anti-cuckoldry tactic, with its occurrence related to a man's suspicions of his partner's sexual infidelity. In two studies, we collected data relevant to both perspectives to explore how these variables interact with men's sexual coercion in an intimate relationship. Regression analyses from Study 1 (self-reports from 256 men) and Study 2 (partner-reports from 290 women) indicated that men's sexual coercion of their partners was consistently predicted by female infidelity and men's controlling behavior, suggesting that both variables are necessary to explain men's sexual coercion. Discussion addressed limitations of the current research and highlighted the importance of integrating multiple levels of analysis when studying men's sexual coercion of their intimate partners. PMID- 18415012 TI - Attraction to sexual violence towards women, sexual abuse of children, and non sexual criminal behavior: testing the specialist vs. generalist models in male college students. AB - A sample of 492 college men anonymously completed an expanded version of the Sexual Experiences Survey, the revised Attraction to Sexual Aggression Scale, and the Marlowe-Crowne Social Desirability Scale Short Form to investigate the relations among perpetration of sexual violence (including rape and sexual assault), attraction to sexual violence, attraction to childhood sexual abuse, and attraction towards other crimes while controlling for the impact of social desirability. Analyses indicated that attractions towards sexual violence, general criminality, and childhood sexual abuse were all significantly interrelated. In addition, sexual assault perpetrators reported higher levels of all three types of attraction as compared to nonperpetrators whereas rape perpetrators reported higher levels of attraction to sexual aggression and criminality. Clinical and research implications are discussed. PMID- 18415013 TI - Breast cancer characteristics at diagnosis and survival among Arab-American women compared to European- and African-American women. AB - BACKGROUND: Data from Arab world studies suggest that Arab women may experience a more aggressive breast cancer phenotype. To investigate this finding, we focused on one of the largest settlements of Arabs and Iraqi Christians (Chaldeans) in the US, metropolitan Detroit- a SEER reporting site since 1973. MATERIALS AND METHODS: We identified a cohort of primary breast cancer cases diagnosed 1973 2003. Using a validated name algorithm, women were identified as being of Arab/Chaldean descent if they had an Arab last or maiden name. We compared characteristics at diagnosis (age, grade, histology, SEER stage, and marker status) and overall survival between Arab-, European-, and African-Americans. RESULTS: The cohort included 1,652 (2%) women of Arab descent, 13,855 (18%) African-American women, and 63,615 (80%) European-American women. There were statistically significant differences between the racial groups for all characteristics at diagnosis. Survival analyses overall and for each SEER stage showed that Arab-American women had the best survival, followed by European American women. African-American women had the poorest overall survival and were 1.37 (95% confidence interval: 1.23-1.52) times more likely to be diagnosed with an aggressive tumor (adjusting for age, grade, marker status, and year of diagnosis). CONCLUSION: Overall, Arab-American women have a distribution of breast cancer histology similar to European-American women. In contrast, the stage, age, and hormone receptor status at diagnosis among Arab-Americans was more similar to African-American women. However, Arab-American women have a better overall survival than even European-American women. PMID- 18415014 TI - Genetic contribution to all cancers: the first demonstration using the model of breast cancers from Poland stratified by age at diagnosis and tumour pathology. AB - The aim of the study is to verify the hypothesis that genetic polymorphisms are associated with the predisposition to all malignancies. Using as a model breast cancers from the homogenous Polish population (West Pomeranian region) after stratification of 977 patients by age at diagnosis (under 51 years and above 50 years) and by tumour pathology (ductal cancers--low and high grade, lobular cancers, ER-positive/negative) we tested this hypothesis. Altogether 20 different groups of breast cancer cases have been analyzed. The results were compared to a group of unaffected controls that were matched by age, sex, ethnicity and geographical location and originated from families without cancers of any site among relatives. Molecular alterations selected for analyses included those which have been previously recognized as being associated with breast cancer predisposition. Statistically significant differences between the breast cancer cases and controls were observed in 19 of the 20 analyzed groups. Genetic changes were present in more than 90% of the breast cancer patients in 18 of 20 groups. The highest proportion of cases with constitutional changes-99.3% (139/140) was observed for lobular cancers. The number and type of genetic marker and/or the level of their association with the specific cancer predisposition was different between groups. Markers associated with majority of groups included: BRCA1, CHEK2, p53, TNRnTT, FGFRnAA, XPD CC/AA and XPD GG. Some markers appeared to be group specific and included polymorphisms in CDKN2A, CYP1B1, M3K nAA, and RS67. PMID- 18415016 TI - Concise synthesis of the pentasaccharide O-antigen of Escherichia coli O83:K24:H31 present in the Colinfant vaccine. AB - A block synthetic approach is presented for the synthesis of the pentasaccharide repeating unit of the O-antigen of E. coli O83:K24:H31 strain, present in the "Colifant" vaccine. The target pentasaccharide has been synthesized by coupling a disaccharide with a trisaccharide in excellent yield. Yields are quite satisfactory in all intermediate steps. PMID- 18415015 TI - Preparation of homogenous oligosaccharide chains from glycosphingolipids. AB - After the discovery of glycosphingolipid (GSL) glycan detaching enzymes, Rhodococcal endoglycoceramidase (EGCase) and leech ceramide glycanase (CGase), the method for enzymatically releasing glycans from GSLs has become the method of choice for preparing intact ceramide-free oligosaccharide chains from GSLs. This paper describes (1) the preparation of the intact oligosaccharides from GM1 (II(3)NeuAcGgOse(4)Cer) and GbOse(4)Cer as examples to show the use of CGase to prepare intact glycan chains from GSLs, and (2) the specificity and detergent requirements of Rhodococcal EGCases for the release of glycan chains from different GSLs. PMID- 18415017 TI - Measuring mechanical properties, including isotonic fatigue, of fast and slow MLC/mIgf-1 transgenic skeletal muscle. AB - Contractile properties of fast-twitch (EDL) and slow-twitch (soleus) skeletal muscles were measured in MLC/mIgf-1 transgenic and wild-type mice. MLC/mIgf-1 mice express the local factor mIgf-1 under the transcriptional control of MLC promoter, selectively activated in fast-twitch muscle fibers. Isolated muscles were studied in vitro in both isometric and isotonic conditions. We used a rapid "ad hoc" testing protocol that measured, in a single procedure, contraction time, tetanic force, Hill's (F-v) curve, power curve and isotonic muscle fatigue. Transgenic soleus muscles did not differ from wild-type with regard to any measured variable. In contrast, transgenic EDL muscles displayed a hypertrophic phenotype, with a mass increase of 29.2% compared to wild-type. Absolute tetanic force increased by 21.5% and absolute maximum power by 34.1%. However, when normalized to muscle cross-sectional area and mass, specific force and normalized power were the same in transgenic and wild-type EDL muscles, revealing that mIgf 1 expression induces a functional hypertrophy without altering fibrotic tissue accumulation. Isotonic fatigue behavior did not differ between transgenic and wild-type muscles, suggesting that the ability of mIgf-1 transgenic muscle to generate a considerable higher absolute power did not affect its resistance to fatigue. PMID- 18415018 TI - An experiment-based model of vein graft remodeling induced by shear stress. AB - Vein graft intimal hyperplasia induced by shear stress is considered to be one of the major causes of vein graft failure. We have developed a mathematical model of vein graft intimal hyperplasia induced by shear stress based on experimental data. Intimal thickness and the rate of intimal thickness change are expressed as functions of shear stress and time. The model coefficients are derived from animal experiments where bilateral rabbit carotid vein grafts are exposed to different shear stress levels. Morphology data of the vein grafts are obtained over multiple time points. The model describes the well-known behavior of intimal thickening, which is inversely related to shear stress. It also depicts the time dependent behavior of vein graft intimal hyperplasia. Finally, the model is used to simulate the intimal growth around a focal stenosis, which was created by ligating the middle of a vein graft using a suture. Simulation results and experimental data agree qualitatively, and demonstrate that the intima thickens more distal to the stenosed area. These experiments establish the potential of the general experiment-based approach for predicting human vein graft remodeling. Other mechanical and biological factors can be included following a similar approach in order to obtain a more accurate vein graft remodeling model. PMID- 18415019 TI - Computer model of nucleotide transport in a realistic porcine aortic trifurcation. AB - Adenosine triphosphate (ATP) is a ubiquitous blood borne agonist which is responsible for the regulation of vascular tone via purinogenic signalling pathways. The present study models the transport of ATP in a realistic porcine aortic trifurcation, which includes multiple branches and bifurcations. The focus of the present study is understanding how pulsatile flow effects mass transfer, observing both mean and transient variations. Unlike in the many idealized models which model transport of low diffusion coefficient species, the realistic geometry leads to very different mass transfer characteristics. These include spiral patterns in the distribution of low concentration fluid. Furthermore, the mean ATP distribution was found to be elevated compared with the steady state; this is attributed to the effects of convective mixing. The results strongly implicate that under certain conditions mass transport in pulsatile flow exhibits different hydrolysis characteristics at the endothelium compared with steady state. Transient variations throughout the the cardiac cycle were found to be small. This small transient response is primarily due to low ATP diffusivity. PMID- 18415020 TI - Fracture risk and bone mineral density in Turner syndrome. AB - Bone health is a major lifelong concern in caring for women and girls with Turner syndrome (TS). There is an approximately 25% increase in fracture risk most of which is related to medium or high impact trauma. The long bones, especially of the forearm are predominantly affected. This fact may be due to a selective cortical bone deficiency in TS which is unrelated to hypogonadism. In addition, lack of adequate estrogen replacement can lead to trabecular bone deficiency and increase in vertebral compression fractures after age 45. Evaluation of bone density by dual X-ray absorptiometry (DEXA) is important, however, it should be used judiciously in TS in view of its inherent tendency to underestimate the bone density of people with short stature. Bone size-independent methods, such as QCT or volumetric transformation of DEXA data should be used in individuals shorter than 150 cm. Achieving optimal bone density is of critical importance for fracture prevention in TS, and should be pursued by timely introduction of hormone replacement therapy, adequate dose of estrogens during the young adult life, optimal calcium and vitamin D intake and regular physical exercise. In addition, other measures to prevent fall and trauma should be considered, including optimizing hearing and vision, avoiding contact sports and exercise to improve coordination. PMID- 18415021 TI - Molecular dynamics simulations of ligand-induced backbone conformational changes in the binding site of the periplasmic lysine-, arginine-, ornithine-binding protein. AB - The periplasmic lysine-, arginine-, ornithine-binding protein (LAOBP) traps its ligands by a large hinge bending movement between two globular domains. The overall geometry of the binding site remains largely unchanged between the open (unliganded) and closed (liganded) forms, with only a small number of residues exhibiting limited movement of their side chains. However, in the case of the ornithine-bound structure, the backbone peptide bond between Asp11 and Thr12 undergoes a large rotation. Molecular dynamics simulations have been used to investigate the origin and mechanism of this backbone movement. Simulations allowing flexibility of a limited region and of the whole binding site, with and without bound ligands, suggest that this conformational change is induced by the binding of ornithine, leading to the stabilisation of an energetically favourable alternative conformation. PMID- 18415022 TI - A molecular mechanism of P-loop pliability of Rho-kinase investigated by molecular dynamic simulation. AB - Rho-kinase is a leading player in the regulation of cytoskeletal events involving smooth muscle contraction and neurite growth-cone collapse and retraction, and is a promising drug target in the treatment of both vascular and neurological disorders. Recent crystal structure of Rho-kinase complexed with a small-molecule inhibitor fasudil has revealed structural details of the ATP-binding site, which represents the target site for the inhibitor, and showed that the conserved phenylalanine on the P-loop occupies the pocket, resulting in an increase of protein-ligand contacts. Thus, the P-loop pliability is considered to play an important role in inhibitor binding affinity and specificity. In this study, we carried out a molecular dynamic simulation for Rho-kinase-fasudil complexes with two different P-loop conformations, i.e., the extended and folded conformations, in order to understand the P-loop pliability and dynamics at atomic level. A PKA fasudil complex was also used for comparison. In the MD simulation, the flip-flop movement of the P-loop conformation starting either from the extended or folded conformation was not able to be observed. However, a significant conformational change in a long loop region covering over the P-loop, and also alteration of ionic interaction-manner of fasudil with acidic residues in the ATP binding site were shown only in the Rho-kinase-fasudil complex with the extended P-loop conformation, while Rho-kinase with the folded P-loop conformation and PKA complexes did not show large fluctuations, suggesting that the Rho-kinase-fasudil complex with the extended P-loop conformation represents a meta-stable state. The information of the P-loop pliability at atomic level obtained in this study could provide valuable clues to designing potent and/or selective inhibitors for Rho kinase. PMID- 18415023 TI - Fast and accurate methods for predicting short-range constraints in protein models. AB - Protein modeling tools utilize many kinds of structural information that may be predicted from amino acid sequence of a target protein or obtained from experiments. Such data provide geometrical constraints in a modeling process. The main aim is to generate the best possible consensus structure. The quality of models strictly depends on the imposed conditions. In this work we present an algorithm, which predicts short-range distances between Calpha atoms as well as a set of short structural fragments that possibly share structural similarity with a query sequence. The only input of the method is a query sequence profile. The algorithm searches for short protein fragments with high sequence similarity. As a result a statistics of distances observed in the similar fragments is returned. The method can be used also as a scoring function or a short-range knowledge based potential based on the computed statistics. PMID- 18415024 TI - Brian MacMahon (1923-2007): founder of modern epidemiology. AB - Brian MacMahon was born in Sheffield, UK in 1923. He served as chair of the Department of Epidemiology at Harvard School of Public Health for more than 30 years. He was admired as a noble and generous man and respected for his shining intellect, scientific integrity, and broad culture. He set the pace for modern epidemiology and led the way for a whole school of epidemiologists who are now spread around the nation and the world. He made major scientific contributions, received several distinguished prizes and awards, and continued to publish insightful papers until the very end. Brian MacMahon was the first editor-in chief of Cancer Causes and Control. PMID- 18415025 TI - Is edema in minimal change disease of childhood really hypovolemic? AB - OBJECTIVES: In this study, we aimed to find out whether children with minimal change disease can be classified as hypervolemic by objective measures. METHODS: Eighteen children with minimal change disease diagnosed at our department between November 2005 and May 2007 were included in this study. All patients were newly diagnosed or relapsed but were steroid free for at least 6 months. In the first week of edema and when edema resolved (5-7 days after initiation of therapy), weight, height and blood pressure were obtained from all patients. Serum and plasma samples were taken following a starvation period of 12-14 h. The volume load of all patients was evaluated, measuring the inferior vena cava indices in each stage by echocardiography. RESULTS: Average weight at presentation was 8.5% higher than the ideal (dry) weight. There were significant differences between the first and post-treatment body weights, abdomen circumference, and systolic and diastolic blood pressure values (P<0.05 for each). The inferior vena cava index (IVCI) values decreased significantly after diuretic treatment (P<0.001), while inferior vena cava collapsibility index (IVCCI) values increased in the post-treatment period (P<0.001). CONCLUSION: We believe that a close follow-up of hypervolemic children with MCD, treated solely with easy-to-handle diuretics instead of I.V. albumin and diuretics may properly solve the edematous state in these patients. PMID- 18415026 TI - Memories: from protein synthesis to photosynthesis. AB - Emphasis herein is on the early years of my scientific life, primarily in graduate school and at the McCollum-Pratt Institute, Johns Hopkins University, as techniques learned and research performed then became the basis for future scientific endeavors. Studies on the mechanism of conversion of light energy into chemical free energy were a logical consequence of earlier investigations on enzyme-catalyzed hydrogen transfer reactions and pyridine nucleotide coenzyme biochemistry. Identification of several protein factors involved in pyridine nucleotide reduction by illuminated chloroplasts is described and, hopefully, adequately and honestly referenced to complementary research in other laboratories. Coupled with progress were changes in nomenclature of the protein factors and are so noted. In particular, David Wharton proposed the descriptive name, ferredoxin, for the non-heme iron and labile sulfide-containing proteins which serve as redox cofactors in a variety of energy conserving reactions. The inclusion of "Lessons" is adapted from Efraim Racker (1976, A new look at mechanisms in bioenergetics. Academic Press, NY). They are lessons that I learned and are included herein solely for graduate students. PMID- 18415027 TI - Genome-wide copy neutral LOH is infrequent in familial and sporadic microsatellite unstable carcinomas. AB - Mismatch repair deficiency in tumors can result from germ line mutations in one of the mismatch repair (MMR) genes (MLH1, MSH2, MSH6 and PMS2), or from sporadic promoter hypermethylation of MLH1. The role of unclassified variants (UVs) in MMR genes is subject to debate. To establish the extend of chromosomal instability and copy neutral loss of heterozygosity (cnLOH), we analyzed 41 archival microsatellite unstable carcinomas, mainly colon cancer, from 23 patients with pathogenic MMR mutations, from eight patients with UVs in one of the MMR genes and 10 cases with MLH1 promoter hypermethylation. We assessed genome wide copy number abnormalities and cnLOH using SNP arrays. SNP arrays overcome the problems of detecting LOH due to instability of polymorphic microsatellite markers. All carcinomas showed relatively few chromosomal aberrations. Also cnLOH was infrequent and in Lynch syndrome carcinomas usually confined to the locus harbouring pathogenic mutations in MLH1, MSH2 or PMS2 In the carcinomas from the MMR-UV carriers such cnLOH was less common and in the carcinomas with MLH1 promoter hypermethylation no cnLOH at MLH1 occurred. MSI-H carcinomas of most MMR UV carriers present on average with more aberrations compared to the carcinomas from pathogenic MMR mutation carriers, suggesting that another possible pathogenic MMR mutation had not been missed. The approach we describe here shows to be an excellent way to study genome-wide cnLOH in archival mismatch repair deficient tumors. PMID- 18415028 TI - Older age and phenformin therapy: a dangerous association. PMID- 18415029 TI - Recurrent appendicitis. PMID- 18415030 TI - Negative regulators in homeostasis of naive peripheral T cells. AB - It is now apparent that naive peripheral T cells are a dynamic population where active processes prevent inappropriate activation while supporting survival. The process of thymic education makes naive peripheral T cells dependent on interactions with self-MHC for survival. However, as these signals can potentially result in inappropriate activation, various non-redundant, intrinsic negative regulatory molecules including Tob, Nfatc2, and Smad3 actively enforce T cell quiescence. Interactions among these pathways are only now coming to light and may include positive or negative crosstalk. In the case of positive crosstalk, self-MHC initiated signals and intrinsic negative regulatory factors may cooperate to dampen T cell activation and sustain peripheral tolerance in a binary fashion (on-off). In the case of negative crosstalk, self-MHC signals may promote survival through partial activation while intrinsic negative regulatory factors act as rheostats to restrain cell cycle entry and prevent T cells from crossing a threshold that would break tolerance. PMID- 18415031 TI - Neurochemical monitoring of therapeutic effects in large human MCA infarction. AB - BACKGROUND AND PURPOSE: Cerebral microdialysis is an invasive monitoring tool allowing analysis of various substances derived from the extracellular space in brain tissue such as glutamate, glycerol, lactate, and pyruvate. In order to assess the potential effects of hemicraniectomy, hypothermia and conservative therapy on these substances, we used neurochemical monitoring with microdialysis in large human stroke patients. METHODS: This is an open, prospective observational study in 24 patients with large MCA infarction undergoing either hypothermia (33 degrees C), hemicraniectomy, or maximum conservative therapy. Microdialysis probe placement was aimed at the peri-infarct tissue within 24 h after stroke onset. Glutamate, glycerol, pyruvate, and lactate were analyzed every 60 min. Measurements of two consecutive days were pooled for statistical analysis. RESULTS: Average glutamate concentrations in patients treated with hemicraniectomy (5.3 +/- 0.5 micromol/l, P < 0.0001; n = 6) and hypothermia (14.5 +/- 3.6 micromol/l, P < 0.0001; n = 14) were significantly lower than in conservatively treated patients (68.3 +/- 5.2 micromol/l; n = 4). Glycerol concentration was significantly lower in patients treated by hypothermia (111 +/- 17 micromol/l; P < 0.0001) and hemicraniectomy (138 +/- 8 micromol/l; P < 0.0001) as compared to conservatively treated patients with 612 +/- 27 micromol/l. The lactate-pyruvate ratio was significantly lower both in the hypothermia (16.2 +/- 3.3) and hemicraniectomy groups (31.3 +/- 1.5) than in the conservative treatment group (56 +/- 2.9). CONCLUSION: Microdialysis allows bed-side monitoring of neuroprotective effects of stroke rescue therapies such as hypothermia and hemicraniectomy. Rescue of peri-infarct tissue and/or prevention of secondary ischemic injury could be associated with a lower mortality in invasively treated patients. PMID- 18415032 TI - Hypothermia for refractory status epilepticus. AB - INTRODUCTION: Status epilepticus (SE) can be refractory to conventional anticonvulsants, requiring anesthetic doses of medications to suppress seizures. This approach carries significant morbidity, is associated with a high fatality rate, and may not always control SE. Hypothermia has been shown to suppress epileptiform activity experimentally, but has not previously been used as a primary modality to control SE in humans. METHODS: Four patients with SE refractory to benzodiazepine and/or barbiturate infusions were treated with hypothermia (target temperature: 31-35 degrees C) using an endovascular cooling system. All received continuous EEG monitoring, three were on midazolam infusions and one had recurrent seizures on weaning from pentobarbital. RESULTS: Therapeutic hypothermia was successful in aborting seizure activity in all four patients, allowing midazolam infusions to be discontinued; three achieved a burst suppression pattern on EEG. After controlled rewarming, two patients remained seizure-free, and all four demonstrated a marked reduction in seizure frequency. Adverse events included shivering, coagulopathy without bleeding, and venous thromboembolism. Two death occurred, neither directly related to hypothermia; however, immunosuppression related to the use of barbiturates and hypothermia may have contributed to an episode of fatal sepsis in one patient. CONCLUSIONS: Hypothermia was able to suppress seizure activity in patients with SE refractory to traditional therapies with minimal morbidity. It appears promising as an alternative or an adjunct to anesthetic doses of other agents, but requires further study to better evaluate its safety and efficacy. PMID- 18415033 TI - Regulated proteolysis of APP and ApoE receptors. AB - The beta-amyloid precursor protein (APP) shares intracellular and extracellular binding partners with the family of receptors for apolipoprotein E (apoE). Binding of APP and apoE receptors to specific extracellular matrix proteins (F spondin and Reelin) promotes their presence on the cell surface and influences whether they will interact with specific cytoplasmic adaptor proteins. Cleavage of APP and apoE receptors at the cell surface occurs by alpha-secretase activities; thus, the processing of these proteins can be regulated by their trafficking either to or from the cell surface. Their cleavages can also be regulated by tissue inhibitor of metalloproteinase-3 (TIMP-3), a metalloprotease inhibitor in the extracellular matrix. ApoE receptors have functions in neuronal migration during development and in proper synaptic function in the adult. Thus, the functions of apoE receptors and by analogy of APP will be modified by the various extracellular and intracellular interactions reviewed in this paper. PMID- 18415035 TI - Addition of rituximab to chop does not increase the risk of cardiotoxicity in patients with non-Hodgkin's lymphoma. AB - BACKGROUND: The addition of rituximab to doxorubicin-containing standard chemotherapy significantly improves response to therapy and reduces the risk of death in B-cell non-Hodgkin's lymphoma (NHL) patients. However, the impact of this approach on doxorubicin-induced cardiotoxicity has not been elucidated. METHODS: Patients who had been planned to receive CHOP or rituximab plus CHOP (R CHOP) combination chemotherapy with a diagnosis of NHL were included in the study. In all patients, systolic and diastolic parameters were measured by using conventional and pulsed-wave tissue Doppler echocardiography, which is more sensitive than conventional lead-dependent techniques, both before and in the sixth month of therapy. RESULTS: There were 28 (M/F; 14/14) patients on CHOP and 33 (M/F; 16/17) patients on R-CHOP. Median age in CHOP and R-CHOP was 49 and 50 years (P = 0.44), respectively. Cumulative doxorubicin doses were 280 and 286 mg/m(2) on CHOP and R-CHOP (P = 0.65), respectively. None of the patients developed clinically evident congestive heart failure. Parameters of systolic function such as LVEF and FS did not significantly change in any patients. In both arms, tissue Doppler parameters of diastolic function such as lateral E and septal E velocity of mitral annulus decreased significantly after therapy (P < 0.001). However, the decrease in diastolic function was similar in both arms (P > 0.05). Conventional Doppler echocardiography yielded consistent findings. CONCLUSION: Both CHOP and R-CHOP cause diastolic dysfunction in the early period following their administration. The addition of rituximab to CHOP chemotherapy does not significantly increase the risk of doxorubicin-induced cardiotoxicity during this period. PMID- 18415036 TI - Transformation of industrialized strain Candida glycerinogenes with resistant gene zeocin via Agrobacterium tumefaciens. AB - Candida glycerinogenes WL2002-5 has a modest sugar tolerance and an extremely high glycerol productivity. Agrobacterium tumefaciens can transfer part of its Ti plasmid, the T-DNA, into the nuclear genome of a wide variety of host cells. In this study, we constructed the plasmid pZR and transferred it into A. tumefaciens LBA4404 to form the strain LBA4404-ZR. LBA4404-ZR was cocultivated with C. glycerologenesis, and putative transformants were identified by selection for zeocin resistance. Polymerase chain reaction and Southern blot analysis confirmed that the gene zeocin was integrated into the genome of engineered C. glycerologenesis. Optimization of the transformation condition was performed in darkness at 25 degrees C on induction medium for 24 h by cocultivation of C. glycerinogenes and LBA4404-ZR with a cell ratio of 1:500-1000. The transformation efficiency reached 2 transformants per 10(4) C. glycerologenesis cells. Our results demonstrated that A. tumefaciens-mediated transformation can be used for C. glycerinogenes. This transformation system can provide the basis for research of C. glycerologenesis in the future. PMID- 18415034 TI - Locomotor dysfunction and pain: the scylla and charybdis of fiber sprouting after spinal cord injury. AB - Injury to the spinal cord (SCI) can produce a constellation of problems including chronic pain, autonomic dysreflexia, and motor dysfunction. Neuroplasticity in the form of fiber sprouting or the lack thereof is an important phenomenon that can contribute to the deleterious effects of SCI. Aberrant sprouting of primary afferent fibers and synaptogenesis within incorrect dorsal horn laminae leads to the development and maintenance of chronic pain as well as autonomic dysreflexia. At the same time, interruption of connections between supraspinal motor control centers and spinal cord output cells, due to lack of successful regenerative sprouting of injured descending fiber tracts, contributes to motor deficits. Similarities in the molecular control of axonal growth of motor and sensory fibers have made the development of cogent therapies difficult. In this study, we discuss recent findings related to the degradation of inhibitory barriers and promotion of sprouting of motor fibers as a strategy for the restoration of motor function and note that this may induce primary afferent fiber sprouting that can contribute to chronic pain. We highlight the importance of careful attentiveness to off-target molecular- and circuit-level modulation of nociceptive processing while moving forward with the development of therapies that will restore motor function after SCI. PMID- 18415037 TI - Incorporating the amino acid properties to predict the significance of missense mutations. AB - Determining if missense mutations are deleterious is critical for the analysis of genes implicated in disease. However, the mutational effects of many missense mutations in databases like the Breast Cancer Information Core are unclassified. Several approaches have emerged recently to determine such mutational effects but none have utilized amino acid property indices. We modified a previously described phylogenetic approach by first classifying benign substitutions based on the assumption that missense mutations that are maintained in orthologs are unlikely to affect function. A consensus conservation score based on 16 amino acid properties was used to characterize the remaining substitutions. This approach was evaluated with experimentally verified T4 lysozyme missense mutations and is shown to be able to sieve out putative biochemical and structurally important residues. The use of amino acid properties can enhance the prediction of biochemical and structurally important residues and thus also predict the significance of missense mutations. PMID- 18415038 TI - Signs of herniosis in women with vaginal prolapse and/or stress incontinence. AB - Gynecological literature pertaining to prolapse or stress urinary incontinence published over the past four decades was reviewed to determine whether signs of herniosis, the systemic connective tissue co-morbidity known to play a significant role in abdominal herniation, were present and differed from controls. A total of eight indications were reported: (1) Genetic factors, i.e., family history and race, were predictive. (2) An increase in the incidence was observed in association with heritable diseases of collagen and their formes frustes (e.g., joint laxity). (3) Recurrence rate after repair was high (30%). (4) Fragmentation and degeneration of smooth muscle and collagen fibers were observed histologically. (5) Biochemistry demonstrated a decline of 24-40% in collagen content of skin, round ligament, cardinal ligament, periurethral vaginal wall, cervix, pubocervical, cervicococcygeal, and vesicovaginal fasciae. (6) In patients with stress urinary incontinence, collagen content decreased 60%. This change was independent of age, parity, menopausal status, and weight. (7) Matrix metalloproteinase (MMP-2 and MMP-9) activity increased fourfold and that of their inhibitor TIMP-1 decreased. (8) Cigarette smoking, an acquired factor, increased the incidence of stress urinary incontinence. This commonality with the etiology of abdominal herniae explains why gynecologists have decreased their emphasis on childbirth injury and, like herniologists, have moved to discard the dogma "prolapse" as a designate for extraperitoneal herniation in the pelvis. PMID- 18415039 TI - Reversible posterior leukoencephalopathy syndrome: a possible manifestation of Wegener's granulomatosis-mediated endothelial injury. AB - We present the case of a 15-year-old girl who had Wegener's granulomatosis with severe intestinal involvement. During the clinical course, she developed generalized seizures and was diagnosed with reversible posterior leukoencephalopathy syndrome (RPLS). Plasma exchange combined with steroid pulse therapy was initiated and showed marked improvement. This is one of the few cases of RPLS without severe hypertension or renal failure, suggesting that RPLS is likely to be a manifestation of Wegener's granulomatosis-mediated endothelial injury. PMID- 18415040 TI - Polyphenolic compounds isolated from the leaves of Myrtus communis. AB - Four hydrolyzable tannins [oenothein B (1), eugeniflorin D(2) (2), and tellimagrandins I (3) and II (4)], two related polyphenolic compounds [gallic acid (5) and quinic acid 3,5-di-O-gallate (6)], and four myricetin glycosides [myricetins 3-O-beta-D-xyloside (7), 3-O-beta-D-galactoside (8), 3-O-beta-D galactoside 6''-O-gallate (9), and 3-O-alpha-L-rhamnoside (10)] were isolated from the leaves of Myrtus communis. Antioxidant activities of the isolated compounds were evaluated by 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging assay. PMID- 18415041 TI - Macroscopic alignment of nanoparticle arrays in soft crystals of cubic and cylindrical polymer micelles. AB - We describe a method to organize nanometer-sized hydrophilic particles into ordered arrays by templating them in the soft, micelle-crystal phases (spherical and cylindrical) of a thermoreversible block copolymer. Small-angle neutron scattering (SANS) with contrast variation is used to show that the dispersed particles (in this case, proteins or silica) form structured arrays by being constrained in the interstitial cavities between the polymer micelles in the ordered micelle crystal. Simple shear is used to macroscopically align both phases of the nanocomposites (micelles and particles) into macro-domains. The temperature-induced order-order transition between templates of spherical and cylindrical micelles is demonstrated as a reversible technique to modify the structure of the templated nanoparticle arrays. PMID- 18415042 TI - Dynamics in shear flow studied by X-ray Photon Correlation Spectroscopy. AB - X-ray Photon Correlation Spectroscopy was used to measure the diffusive dynamics of colloidal particles in a shear flow. The results presented here show how the intensity autocorrelation functions measure both the diffusive dynamics of the particles and their flow-induced, convective motion. However, in the limit of low flow/shear rates, it is possible to obtain the diffusive component of the dynamics, which makes the method suitable for the study of the dynamical properties of a large class of complex soft-matter and biological fluids. An important benefit of this experimental strategy over more traditional X-ray methods is the minimization of X-ray-induced beam damage. While the method can be applied also for photon correlation spectroscopy in the visible domain, our analysis shows that the experimental conditions under which it is possible to measure the diffusive dynamics are easier to achieve at higher q values (with X rays). PMID- 18415043 TI - Ultrasound velocimetry in a shear-thickening wormlike micellar solution: evidence for the coexistence of radial and vorticity shear bands. AB - We carried out pointwise local velocity measurements on 40 mM cetylpyridinium chloride-sodium salicylate (CPyCl-NaSal) wormlike micellar solution using high frequency ultrasound velocimetry in a Couette shear cell. The studied wormlike solution exhibits Newtonian, shear-thinning and shear-thickening rheological behavior in a stress-controlled environment. Previous rheology, flow visualization and small-angle light/neutron scattering experiments in the shear thickening regime of this system showed the presence of stress-driven alternating transparent and turbid rings or vorticity bands along the axis of the Couette geometry. Through local velocity measurements we observe a homogeneous flow inside the 1mm gap of the Couette cell in the shear-thinning (stress-plateau) region. Only when the solution is sheared beyond the critical shear stress (shear thickening regime) in a stress-controlled experiment, we observe inhomogeneous flow characterized by radial or velocity gradient shear bands with a highly sheared band near the rotor and a weakly sheared band near the stator of the Couette geometry. Furthermore, fast measurements performed in the shear thickening regime to capture the temporal evolution of local velocities indicate coexistence of both radial and vorticity shear bands. However the same measurements carried out in shear rate controlled mode of the rheometer do not show such rheological complexity. PMID- 18415045 TI - Blockage of the STAT3 signaling pathway with a decoy oligonucleotide suppresses growth of human malignant glioma cells. AB - Gliomas are the most common type of primary tumor in the human central nervous system. STAT3, a signal transducer and activator of transcription 3, is over expressed in gliomas. Its involvement in tumorgenesis can be attributed to its ability to induce cell proliferation and inhibit apoptosis. Double-stranded decoy oligodeoxynucleotides (ODNs) which correspond closely to the STAT3 response element within the c-fos promoter are a potential tool for inhibiting a variety of tumor cell growth. To investigate its therapeutic potential in malignant gliomas, a 15-mer double-stranded decoy ODN mimicking STAT3-specific cis-elements was transfected into two glioma cell lines, U251 and A172. The STAT3 decoy ODN treatment specifically blocked STAT3 signaling and subsequently inhibited U251 and A172 cell proliferation by inducing apoptosis and cell-cycle arrest. The ODN treatment also decreased transcription and translation of downstream STAT3 target genes including c-myc, cyclin D1 and bcl-xl in both cell lines. Thus, targeted blockade of the STAT3 signaling pathway with a decoy ODN is a potential anti glioma therapeutic approach. PMID- 18415044 TI - Activities of DNA-PK and Ku86, but not Ku70, may predict sensitivity to cisplatin in human gliomas. AB - OBJECTIVE: This study was designed to investigate the relationship between activities of DNA-dependent protein kinase (DNA-PK), its subunits Ku86/Ku70, and sensitivities to cisplatin in human glioma samples. METHODS: Thirty-six glioma samples from patients without prior treatment before neurosurgery were included in this study. The sensitivities to cisplatin as indicated by IC(50) (the inhibitory concentration leading to 50% cell death) were assessed by 3-[4,5 dimethylthiazol-2-yl]-2,5-diphenytetrazolium (MTT) assay; activities of DNA-PK and Ku70/Ku86 were analyzed by SigmaTECT DNA-Dependent Protein Kinase Assay System and Ku70/Ku86 DNA Repair Kit, respectively. RESULTS: Sensitivities to cisplatin correlated with the activities of DNA-PK/Ku86, but not with the Ku70 or other clinical parameters such as age, sex of the patients, pathological gradings of the tumors, or tumor size. The levels of DNA-PK activities also associated with pathological grading and Ku86, but not with other clinical parameters. The tumors of the patients who failed to respond to cisplatin-based chemotherapy tended to display higher activity levels of DNA-PK and Ku86. Furthermore, platinum-based chemotherapy did not result in significant changes of DNA-PK/Ku activities in four matched samples before and after chemotherapy. CONCLUSION: Pretreatment determination of DNA-PK/Ku86 activities might be helpful in identifying patients who will actually benefit from platinum-based treatment. PMID- 18415046 TI - Outcome of pediatric pineoblastoma after surgery, radiation and chemotherapy. AB - INTRODUCTION: Pineoblastomas are a category of supratentorial primitive neuroectodermal tumors (sPNETs) occurring in the pineal gland; some studies support the impression that patients with pineoblastomas have a worse prognosis than those with other sPNETs. METHODS: We reviewed the medical records and tissue sections of all patients with the diagnosis of pineoblastoma that were treated at the Dana-Farber Cancer Institute/Children's Hospital Boston Pediatric Brain Tumor Program between 1986 and 2005. RESULTS: Thirteen patients with the pathologic diagnosis of pineoblastoma were treated at our Hospital; 11 of these cases had complete records suitable for study. The median age was 8 years 8 months (5 F, 6 M). Surgical, radiation and chemotherapeutic regimens varied from case to case. Three patients had gross total resection and are alive and free of disease, versus four of eight with subtotal resection or biopsy only. Patients who received CSI and multi-agent chemotherapy had improved overall survival. CONCLUSIONS: Seven of eleven patients with pineoblastoma are currently alive and free of disease, reflecting an improved outcome and longer survival than previously appreciated. Gross total surgical resection appeared to correlate with improved survival, as did treatment with craniospinal irradiation and multi-agent chemotherapy. Further study of this group of patients as a distinct diagnostic entity will be necessary to determine optimal therapy. PMID- 18415047 TI - Combination antifungal therapy involving amphotericin B, rapamycin and 5 fluorocytosine using PEG-phospholipid micelles. AB - PURPOSE: Rapamycin and 5-fluorocytosine (5-FC) are antifungal agents with unique mechanisms of activity, with potential for cooperative interaction with AmB. Combination antifungal therapy involving conventional AmB has been restricted by poor physical stability and compatibility with antifungal drugs and vehicles. METHODS: AmB and rapamycin were encapsulated in 1,2-distearoyl-sn-glycero-3 phosphoethanolamine-N-methoxy poly(ethylene glycol) (PEG-DSPE) micelles using a solvent evaporation method. The physical stability of micelle encapsulated AmB and rapamycin with 5-FC and saline was evaluated using dynamic light scattering (DLS). In vitro susceptibility of Candida albicans isolates to 5-FC and PEG-DSPE micelle solubilized AmB and rapamycin has been evaluated. Interactive effects have been quantified using a checkerboard layout. RESULTS: In contrast with conventional AmB, PEG-DSPE micelles encapsulating AmB and rapamycin are compatible with saline and 5-FC over 12 h. The solubilized drugs retain high level of potency in vitro. The combination of solubilized AmB and rapamycin was indifferent, as fractional inhibitory concentration (FIC) index and combination index (CI) values were approximately 1. Combinations of solubilized AmB or rapamycin with 5-FC, and the three-drug combination were moderately synergistic since the FIC index and CI values were consistent less than 1. CONCLUSIONS: These results indicate that AmB solubilized in PEG-DSPE micelles is compatible with solubilized rapamycin and 5-FC. The indifferent or moderately synergistic activity of combinations is encouraging and warrants further investigation in appropriate rodent models. PMID- 18415048 TI - Emerging non-invasive Raman methods in process control and forensic applications. AB - This article reviews emerging Raman techniques (Spatially Offset and Transmission Raman Spectroscopy) for non-invasive, sub-surface probing in process control and forensic applications. New capabilities offered by these methods are discussed and several application examples are given including the non-invasive detection of counterfeit drugs through blister packs and opaque plastic bottles and the rapid quantitative analysis of the bulk content of pharmaceutical tablets and capsules without sub-sampling. PMID- 18415049 TI - New predictive models for blood-brain barrier permeability of drug-like molecules. AB - PURPOSE: The goals of the present study were to apply a generalized regression model and support vector machine (SVM) models with Shape Signatures descriptors, to the domain of blood-brain barrier (BBB) modeling. MATERIALS AND METHODS: The Shape Signatures method is a novel computational tool that was used to generate molecular descriptors utilized with the SVM classification technique with various BBB datasets. For comparison purposes we have created a generalized linear regression model with eight MOE descriptors and these same descriptors were also used to create SVM models. RESULTS: The generalized regression model was tested on 100 molecules not in the model and resulted in a correlation r2 = 0.65. SVM models with MOE descriptors were superior to regression models, while Shape Signatures SVM models were comparable or better than those with MOE descriptors. The best 2D shape signature models had 10-fold cross validation prediction accuracy between 80-83% and leave-20%-out testing prediction accuracy between 80 82% as well as correctly predicting 84% of BBB+ compounds (n = 95) in an external database of drugs. CONCLUSIONS: Our data indicate that Shape Signatures descriptors can be used with SVM and these models may have utility for predicting blood-brain barrier permeation in drug discovery. PMID- 18415050 TI - The asymmetry of the unstirred water layer in permeability experiments. AB - PURPOSE: To elucidate the apical and basolateral components of the total unstirred water layer in regular permeability experiment. METHODS: A novel stirring apparatus was constructed to remove the basolateral unstirred water layer. Caco-2 cells were used as the permeability barrier both in Transwell-type and side-by-side apparatuses. Permeability experiments were done with several ionisable compounds at various pH and stirring conditions. The permeabilities of the cell monolayer, the unstirred water layer and the polycarbonate filter were calculated either from experimental data or theoretically. RESULTS: The unstirred water layer was thicker in the Transwell apparatus than in the side-by-side chamber even in the presence of vigorous basolateral magnetic stirring. Calculations indicated that the apical unstirred water layer is thicker than the basolateral layer. Different cellular permeability coefficients were obtained from the two permeability apparatuses. CONCLUSIONS: An orbital shaker does not produce symmetric hydrodynamics in both chambers of Transwell apparatus. The asymmetric unstirred water layer may complicate the exact analysis of polarized transport. PMID- 18415051 TI - Thermal profiling for parallel on-line monitoring of biomass growth in miniature stirred bioreactors. AB - Recently we have described the design and operation of a miniature bioreactor system in which 4-16 fermentations can be performed (Gill et al., Biochem Eng J 39:164-176, 2008). Here we report on the use of thermal profiling techniques for parallel on-line monitoring of cell growth in these bioreactors based on the natural heat generated by microbial culture. Results show that the integrated heat profile during E. coli TOP10 pQR239 fermentations followed the same pattern as off-line optical density (OD) measurements. The maximum specific growth rates calculated from off-line OD and on-line thermal profiling data were in good agreement, at 0.66+/-0.04 and 0.69+/-0.05 h(-1) respectively. The combination of a parallel miniature bioreactor system with a non-invasive on-line technique for estimation of culture kinetic parameters provides a valuable approach for the rapid optimisation of microbial fermentation processes. PMID- 18415053 TI - Charles Sugrue, M.D., of Cork (1775-1816) and the first description of a classical medical condition: phaeochromocytoma. PMID- 18415052 TI - Expression and secretion of recombinant ZZ-EGFP fusion protein by the methylotrophic yeast Pichia pastoris. AB - We constructed a fusion protein ZZ-EGFP by fusing the ZZ domains of staphylococcal protein A (SpA) and enhanced green fluorescent protein (EGFP). ZZ EGFP was secreted in the yeast, Pichia pastoris, with a hexahistidine tag. Its expression level was determined by measuring the fluorescence of EGFP. When the recombinant yeast cells in shake-flasks were induced with 0.5% methanol for 96 h, a maximum yield of 115 mg ZZ-EGFP/l was obtained. The resulting ZZ-EGFP fusion protein retained immunoglobulin G (IgG)-binding capacity and EGFP fluorescence. ZZ-EGFP was then used in immunofluorescence assays for detecting antinuclear antibodies (ANA); it produced a good signal that was comparable in its brightness and fluorescence pattern to that generated with fluorescein isothiocyanate (FITC) labelled anti-human IgG. Thus, ZZ-EGFP showed great potential in immunological applications due to its ability to bind to various IgG from different animal sources. PMID- 18415054 TI - Construction of a recombinant S. cerevisiae expressing a fusion protein and study on the effect of converting xylose and glucose to ethanol. AB - Gene XYL1 from Candida shehatae and gene XYL2 from Pichia stipitis were amplified by polymerase chain reaction (PCR), and the two genes were both placed under the strong promoter of alcohol dehydrogenase (ADH) of plasmid pAD2 to produce the recombinant expression vector pAD2-P12. Because the amplified XYL1 fragment lacks the stop codon UAA, the polypeptide expressed in yeast cells should be a fusion protein, which is a fusion of xylose reductase and xylitol dehydrogenase. Subsequently, the pAD2-P12 vector was transformed into Saccharomyces cerevisiae YS58 to produce a recombinant S. cerevisiae YS58-12. It was indicated that S. cerevisiae YS58-12 has the ability of metabolizing xylose to produce ethanol by fermentation experiment. The result of cofermentation of glucose and xylose by using this recombinant S. cerevisiae YS58-12 showed a relatively satisfactory result. The highest percentage of xylose consumption rate reached 81.3% and the ethanol yield was equal to 67.14% of the ideal value. PMID- 18415056 TI - [Abstracts DEGRO-OGRO 2008. Abstracts of the 14th Annual Congress of the German Society for Radiation Oncology and the 25th Annual Congress of the Austrian Society for Radiation Oncology, Radiobiology and Medical Radiation Physics. May 1 4, 2008. Vienna, Austria]. PMID- 18415055 TI - Characterization of excitability and voltage-gated ion channels of neural progenitor cells in rat hippocampus. AB - We have observed the excitability of rat neural progenitor cells (NPCs) and characterized the profile of the voltage-gated ion channels, with the help of the electrophysiological measurements in NPCs in vitro, in order to clarify the electrophysiological property of NPCs. The membrane potential changes of neural progenitors were detected with fluorescent dye bis-(1,3-dibutylbarbituric acid) trimethine oxonol (DiBAC4(3)) by a confocal laser scanning microscope. The changes of fluorescent intensity of DiBAC4(3) stain after KCl stimulation were slight which indicated that the NPCs were inexcitable. Under the current-clamp, spontaneous discharge was not detected and the action potential was failed to be elicited. These findings were consistent with the result from DiBAC4(3) staining. Under the voltage clamp, the NPCs expressed two types of outward K(+) currents with no evidence for Na(+) currents. An outward delayed rectifier type K(+) current and outward transient K(+) current were elicited. Our findings demonstrate NPCs' electrophysiological properties: the electrical inexcitability indicated by the presence of two types of K(+) currents and the absence of Na(+) current. PMID- 18415057 TI - [Abstracts of the 5th Brachytherapy Symposium at the DEGO, OGRO, SASRO meeting with workshop. September 6-8, 2007. Linz, Austria]. PMID- 18415058 TI - [Abstracts OGRO 2007. Abstracts of the 24th Annual Meeting of the Austrian Society for Radiation Oncology, Radiobiology, and Medical Radiation Physics. October 17-20, 2007. Graz, Austria]. PMID- 18415059 TI - Acute necrotizing encephalopathy of childhood: typical findings in an atypical disease. AB - Acute necrotizing encephalopathy of childhood (ANEC) is a disease entity seen nearly exclusively in East Asian children that is characterized by multifocal, symmetric lesions involving the thalami, brainstem, cerebellum, and white matter. We present a child who developed dramatic neurologic symptoms following a viral prodrome. Serial MRI examinations demonstrated characteristic lesions of ANEC, while laboratory analyses revealed evidence of acute infection with human herpesvirus-6 (HHV-6). We highlight the MRI findings in both the acute and convalescent phases of ANEC, discuss the implications of neuroimaging on the child's clinical course, and emphasize the integral role of the radiologist in correctly diagnosing this rare disease. PMID- 18415060 TI - The impact of herbivore-induced plant volatiles on parasitoid foraging success: a general deterministic model. AB - Parasitoids respond to volatiles that plants produce when injured by herbivores. A considerable body of literature addresses the chemical pathways of herbivore induced volatile production. However, there is almost no theory or data on how timing of volatile release in relationship to host availability for parasitization impacts the utility of these cues to parasitoids and on the extent that this volatile release timing might increase or decrease the percent of herbivores that become parasitized. This kind of information is critical in judging the benefits that might accrue from a breeding program aimed at enhancing herbivore-responsive volatile production. We developed a general model to begin examining this issue by using available parameters from two tritrophic systems. The model uses herbivore oviposition, development, and mortality rates, linked to a range of plant volatile induction and cessation periods for calculating the proportion of plants in a field that are (1) not producing volatiles but occupied by suitable herbivore hosts, (2) producing volatiles and occupied by suitable herbivore hosts, (3) producing volatiles but not occupied by suitable herbivore hosts, and (4) not producing volatiles and not occupied by suitable herbivore hosts. The impact of the plant volatiles on parasitoid foraging success is then determined by comparing the expected number of hosts parasitized when the parasitoid focuses solely on the volatile-producing plants to when it forages randomly among all plants. Under some conditions, parasitoids can attack three times more herbivores if they focus on volatile-producing plants. However, when we simulate plants that take several days to cease volatile production after pupation or death of the herbivore, parasitization rate does not increase when parasitoids use volatiles as cues. The utility of the volatile cues is consistently greater when a smaller proportion of plants is occupied by herbivores, indicating that their usefulness may be reduced to zero in fields saturated with volatiles. PMID- 18415061 TI - The chemistry of the postpharyngeal gland of female European beewolves. AB - Females of the European beewolf, Philanthus triangulum, possess a large glove shaped gland in the head, the postpharyngeal gland (PPG). They apply the content of the PPG to their prey, paralyzed honeybees, where it delays fungal infestation. Here, we describe the chemical composition of the gland by using combined GC-MS, GC-FTIR, and derivatization. The PPG of beewolves contains mainly long-chain unsaturated hydrocarbons (C23-C33), lower amounts of saturated hydrocarbons (C14-C33), and minor amounts of methyl-branched hydrocarbons (C17 C31). Additionally, the hexane-soluble gland content is comprised of small amounts of an unsaturated C25 alcohol, an unknown sesquiterpene, an octadecenylmethylester, and several long-chain saturated (C25, C27) and unsaturated (C23-C27) ketones, some of which have not yet been reported as natural products. Surprisingly, we found a dimorphism with regard to the major component of the PPG with some females having (Z)-9-pentacosene, whereas others have (Z)-9-heptacosene as their predominant component. The biological relevance of the compounds for the prevention of fungal growth on the prey and the significance of the chemical dimorphism are discussed. PMID- 18415062 TI - [Cutaneous vasculitides. A diagnostic approach]. AB - Vasculitis is characterized by an inflammatory reaction of vessel walls with damage to the dependent tissues. Forms of vasculitis which frequently have skin changes include leukocytoclastic angiitis (LcV), Henoch-Schonlein purpura (HSP), cutaneous polyarteriitis nodosum (cPAN), erythema elevatum et diutinum (EED) and urticarial vasculitis (UV). In other forms of vasculitis, systemic manifestations predominate but there are a variety of skin changes. Kawasaki disease (MK), cryoglobulinemic vasculitis (kV), Wegener granulomatosis (WG), Churg-Strauss syndrome (CSS) and microscopic polyangitis (MPA) belong to this group. The causes of vasculitis are heterogeneous. Triggers include infections, drugs, collagen vascular diseases, autoimmune diseases and lymphoproliferative disorders. Idiopathic vasculitis, particularly LcV and EED, occur only once and have a self limited course. The diagnostic work up depends on the clinical picture and includes inflammatory markers, circulating immune complexes, different types of cryoglobulins and anti-neutrophilic cytoplasmic antibodies, collagen vascular disease specific autoantibodies and additional hematological studies. Vasculitis can manifest in many organs and requires a thorough work up specifically in cases where WG, MPA, CSS and PAN are under consideration. PMID- 18415063 TI - [Histology of cutaneous vasculitides]. AB - Identification of vasculitis by skin biopsy represents the diagnostic gold standard. Skin biopsies should be taken from fresh lesions or from margin of an ulceration and should contain all layers of the skin including subcutis. Classification of vasculitis is based on histological criteria considering the size of the predominantly affected vessel, the distribution of vasculitis in the dermis and subcutis, and the predominant inflammatory cell-type. In cutaneous vasculitis, small and medium-sized vessels of the arterial and/or venous system are predominant affected. Vasculitis of the larger-sized blood vessels is based on inflammatory cells within the wall of the vessel; in small vessel vasculitis, additional features include fibrin within the vessel wall and/or an intraluminal thrombus and/or perivascular and interstitial infiltrates of neutrophils, nuclear dust and extravasated erythrocytes are required for the diagnosis of vasculitis. Leukocytoclastic vasculitis is the most common form of cutaneous vasculitis. An correct diagnosis requires careful correlation of medical history, the clinical, serological, imaging and direct immunofluorescence data, and histologic findings. PMID- 18415064 TI - [Photodynamic therapy using m-THPC (Foscan). Treatment of head and neck squamous cell carcinoma]. PMID- 18415066 TI - [Otorhinolaryngologic disorders associated with diving]. AB - The number of Germans practicing sea diving has grown, so doctors are increasingly confronted with questions pertaining to diving. They concern not only patient fitness but also prevention and therapy of associated disorders and disease. This article describes predisposing factors and otologic, rhinologic, and laryngologic problems associated with diving. The most common medical problem is inflammation of the external auditory canal; the most common -Barotrauma is to the middle ear. Case studies are used to demonstrate these and other disorders associated with diving. PMID- 18415065 TI - [Fitness to dive in the otorhinolaryngological field]. AB - In line with the rising number of recreational divers, the otorhinolaryngologist has to deal with growing numbers of diving-associated disorders of the ear, nose and throat (ENT). Nevertheless, the majority of divers present to their ENT doctor for assessment of their fitness to dive. On the basis of long-term follow up examinations and increasing experience in diving medicine, even divers with a history of ENT problems can be considered fit to dive. Therefore, diving is possible after tympanoplasty, surgery to improve hearing including stapesplasty, after implantation of middle ear amplifiers or cochlear implants, after sinus or scull base surgery and even after canal wall down mastoidectomy, provided that certain requirements are fulfilled. Assessing fitness to dive after inner ear barotrauma as well as after inner ear decompression illness requires meticulous consideration of residual damage and possible underlying conditions like vascular right-to-left shunts. This article is based on the new recommendations of the German Undersea and Hyperbaric Medical Society for the assessment of fitness to dive in the otorhinolaryngological field. PMID- 18415067 TI - [Diving medicine. ENT specialists' expertise needed and required]. PMID- 18415068 TI - [No correlation between reflux of gastric contents and obstructive sleep apnea]. AB - Reflux of gastric contents can be associated with many ENT disorders. Some authors describe an association with obstructive sleep apnea, but it is not clear whether the reflux causes the apnea or vice versa. Furthermore, authors did not distinguish between gastroesophageal reflux and extraesophageal reflux, which is essential to understand a connection with obstructive sleep apnea. Therefore, we performed polysomnography and two-channel pH testing simultaneously in patients with obstructive sleep apnea syndrome (OSAS) and compared the findings with data of healthy volunteers. After exclusion of a changed sleep architecture due to the pH testing system, the results show that patients with OSAS do not suffer more often from reflux than healthy volunteers. This is true for gastroesophageal as well as for extraesophageal reflux. Furthermore, an increasing number of reflux events during the night is not correlated with the number of apnea events. Thus, in summary our data cannot support the postulation that there is a connection between obstructive sleep apnea syndrome and reflux disease. PMID- 18415069 TI - [Assessing the progress of grammar acquisition in young children]. AB - Children need to acquire approximately 50 words within their first 24 months. This enables them to start combining words to make short sentences, which is also the start of learning rules governing grammar. When children start to construct sentences, the mean length of their utterances correlates with their implicit grammar knowledge. Therefore, the mean length of utterance estimation can be used to assess and monitor the progress of grammar acquisition. PMID- 18415070 TI - miRNAs at the heart of the matter. AB - Cardiovascular disease is among the main causes of morbidity and mortality in developed countries. The pathological process of the heart is associated with altered expression profile of genes that are important for cardiac function. MicroRNAs (miRNAs) have emerged as one of the central players of gene expression regulation. The implications of miRNAs in the pathological process of cardiovascular system have recently been recognized, representing the most rapidly evolving research field. Here, we summarize and analyze the currently available data from our own laboratory and other groups, providing a comprehensive overview of miRNA function in the heart, including a brief introduction of miRNA biology, expression profile of miRNAs in cardiac tissue, role of miRNAs in cardiac hypertrophy and heart failure, the arrhythmogenic potential of miRNAs, the involvement of miRNAs in vascular angiogenesis, and regulation of cardiomyocyte apoptosis by miRNAs. The target genes and signaling pathways linking the miRNAs to cardiovascular disease are highlighted. The applications of miRNA interference technologies for manipulating miRNA expression, stability, and function as new strategies for molecular therapy of human disease are evaluated. Finally, some specific issues related to future directions of the research on miRNAs relevant to cardiovascular disease are pinpointed and speculated. PMID- 18415071 TI - [Functional results after temporary continuous drainage of the hypocontractile bladder. The potential rehabilitation of the detrusor]. AB - BACKGROUND: Impaired bladder emptying is a common problem in older people and a challenging task in treatment. Conservative and medical treatment options have shown beneficial effects on micturition; however, in a substantial number of patients the effectiveness of these therapies is disappointing. In the end the decompensated bladder needs indwelling catheterisation. To study the effects on the detrusor function, we analysed the urodynamic data of 31 patients during long term bladder drainage retrospectively. PATIENTS AND METHODS: All 17 female and 14 male patients showed impaired detrusor contractility, enlarged bladder capacity, decreased sensitivity and a high post-void residual urine volume (PVR). After exclusion of an acute pathology, the patients were treated continuously with a suprapubic catheter for an average of 13.1 weeks. By urodynamic measurements before and after the drainage period, we analysed the filling parameters, pressure-flow patterns, PVR and detrusor contractility. RESULTS: At the end of the drainage period, significant changes in the detrusor function were obvious. Compared with the pre-treatment situation, the bladder volume at first desire to void decreased from 306.92 ml to 281.7 ml and the maximum bladder capacity from 691.8 ml to 496.8 ml, respectively. The compliance of the detrusor muscle diminished in the same period of time from 65.6 ml/cmH2O to 51.8 ml/cmH2O. The PVR dropped by 227.2 ml in average. The maximum flow rate was 9.4 ml/s, and the maximum detrusor pressure increased slightly up to 23.6 cmH2O. CONCLUSION: The continuous drainage of the bladder results in significant changes in the motoric as well as sensoric detrusor function. The reduced bladder capacity and the decreased PVR might be indications of a regenerating process of the detrusor. The long-term drainage of the bladder shows beneficial and therefore therapeutic effects. It still remains to be investigated on a functional as well as structural basis to what extent age, gender and pathogenesis influences the rehabilitation of the detrusor. PMID- 18415072 TI - Expression of a Bacillus thuringiensis toxin (cry1Ab) gene in cabbage (Brassica oleracea L. var. capitata L.) chloroplasts confers high insecticidal efficacy against Plutella xylostella. AB - Chloroplast genetic engineering is an environmentally friendly approach, where the foreign integrated gene is often expressed at a higher level than nuclear transformation. The cry1Ab gene was successfully transferred into the cabbage chloroplast genome in this study. The aadA and cry1Ab genes were inserted into the pASCC201 vector and driven by the prrn promoter. The cabbage-specific plastid vectors were transferred into the chloroplasts of cabbage via particle gun mediated transformation. Regenerated plantlets were selected by their resistance to spectinomycin and streptomycin. According to antibiotic selection, the regeneration percentage of the two cabbage cultivars was 4-5%. The results of PCR, Southern, Northern hybridization and western analyses indicated that the aadA and cry1Ab genes were not only successfully integrated into the chloroplast genome, but functionally expressed at the mRNA and protein level. Expression of Cry1Ab protein was detected in the range of 4.8-11.1% of total soluble protein in transgenic mature leaves of the two species. Insecticidal effects on Plutella xylostella were also demonstrated in cry1Ab transformed cabbage. The objectives of this study were to establish a gene transformation system for Brassica chloroplasts, and to study the possibility for insect-resistance in dicot vegetables using chloroplast gene transformation. PMID- 18415073 TI - The relationship between admission blood glucose levels and hospital mortality. AB - AIMS/HYPOTHESIS: The purpose of this study was to examine the relationship between blood glucose level (BGL) on admission with mortality rates among patients admitted to hospital through the Emergency Department. METHODS: In a prospective observational study, BGLs were routinely measured on 6,187 consecutive patients requiring blood testing and admitted through the Emergency Department of a tertiary referral hospital. These measurements were matched against demographic data and hospital mortality rates. RESULTS: Overall in hospital mortality was 4.8%. Admission BGL was an independent predictor of mortality (HR 1.04 per 1 mmol/l increase, 95% CI 1.02-1.06, p=0.02). There was a significant interaction between diabetes status and increasing BGL on mortality (p<0.001), with higher BGLs being associated with greater mortality among non diabetic than among diabetic patients. Among non-diabetic patients, the lowest mortality rate (3.0%) was in people with a BGL of 4.0-5.9 mmol/l. Compared with this group, patients with a BGL of 8.0-9.9 mmol/l had increased mortality rate (7.6%, HR 1.56, 95% CI 1.03-2.35, p=0.04, after adjustment for age and sex). The risk increased further at higher glucose levels. In the cohort with diagnosed diabetes, the increase in mortality rates at higher BGL bands was not significant. CONCLUSIONS/INTERPRETATION: Among people who do not have diabetes, even modest degrees of hyperglycaemia on hospital admission are associated with increased mortality. PMID- 18415074 TI - [Peroneal nerve palsy after total knee arthroplasty under continuous epidural anaesthesia]. AB - BACKGROUND: Peroneal nerve palsy is a rare but distressing complication of total knee arthroplasty (TKA). After introducing a standardised intraoperative and postoperative epidural anaesthesia protocol under otherwise unchanged perioperative management, we noted a sudden cumulation of peroneal nerve palsies after TKA. PATIENTS AND METHODS: In this retrospective study we checked the patients' histories for well-known risk factors for nerve lesions after TKA as well as for those risk factors controversially discussed in the literature. RESULTS: We found an additive harmful impact of epidural anaesthesia leading to unrecognised pressure on the peroneal nerve, which caused, in combination with a pressure lesion of the pneumatic tourniquet, an axonal lesion in terms of a double-crush syndrome. By lowering the pneumatic tourniquet pressure and carefully positioning the operated leg, we found a clearly reduced risk of nerve lesion while preserving the advantages of epidural anaesthesia for the patient. CONCLUSION: To prevent a peroneal lesion after TKA while using continuous epidural anaesthesia, we strongly recommend limiting the pneumatic tourniquet pressure to 320 mmHg while ensuring pressure-free positioning of the operated leg. PMID- 18415075 TI - [Whiplash-associated disorders: a challenge for the expert in compensation claims and litigation]. AB - In the literature, the association between chronic complaints of the cervical spine and sustained"whiplash trauma" is a matter of controversy. Whiplash trauma is identified as contusion, compression, and/or sprain without definite imaging proof of injury to the bone, disk, or ligaments.However, complex chronic complaints leading to permanent and significant functional limitations in daily living and work may develop. Besides the diagnostic and therapeutic challenges for the treating physician, the expert confronted with compensation claims very often must decide whether whiplash trauma of the spine may lead to significant and permanent functional limitations or loss with chronic disorders. The data from the literature dealing with this topic are contradictory and deficient. In this article, based on the present available data from the literature, we critically examine the causality between spinal whiplash trauma and possible permanent chronic complaints in order to support the process of decision making in cases of litigation and controversial compensation claims. PMID- 18415076 TI - [Oral anticoagulation after major hip or knee replacement surgery: a process driven managerial pharmacoeconomic analysis in German hospitals]. AB - BACKGROUND: The thrombin inhibitor dabigatranetexilat is used for prophylaxis of venous thromboembolism after total hip or knee replacement surgery (THR/TKR). Patients can take it orally in hospitals. METHOD: In a managerial pharmacoeconomic analysis of six German acute-care hospitals and six German rehabilitation hospitals, the use of dabigatranetexilat was compared with the use of low-molecular-weight heparins. RESULTS: The analysis showed that the new drug led to an economic advantage for an acute-care hospital of 2.43 euro per patient per day. In a rehabilitation hospital, the use of dabigatranetexilat led to an economic advantage of 1.40 euro per patient per day. CONCLUSION: These results have direct implications for drug decisions in hospitals. To demonstrate that fact, the price difference between dabigatranetexilat and low-molecular-weight heparins was derived to lead exactly to their"economic neutrality" from the hospital's point of view. PMID- 18415078 TI - Comment on "Surviving Sepsis Campaign: International guidelines for management of severe sepsis and septic shock: 2008" by Dellinger et al. PMID- 18415079 TI - Insights into the NrpR regulon in Methanosarcina mazei Go1. AB - The methanogenic archaeon Methanosarcina mazei strain Go1 contains two homologues of NrpR, the transcriptional repressor of nitrogen assimilation genes recently discovered and characterized in Methanococcus maripaludis. Insertion of a puromycin-resistance conferring cassette into MM1085 encoding a single NrpR domain with an N-terminal helix-turn-helix domain (NrpRI) lead to a significant reduction of the lag-phase after a shift from nitrogen sufficiency to nitrogen limitation. Consistent with this finding, loss of NrpRI resulted in significantly increased transcript levels of genes involved in nitrogen fixation or nitrogen assimilation though growing under nitrogen sufficiency as demonstrated by quantitative reverse transcriptional PCR analysis. Genome-wide analysis using DNA microarrays confirmed that transcript levels of 27 ORFs were significantly elevated in the M. mazei MM1085::pac mutant under nitrogen sufficiency, including genes known to be up-regulated under nitrogen limitation (e.g., nifH, glnA(1), glnK(1)), and 17 additional genes involved in metabolism (4), encoding a flagella related protein (1) and genes encoding hypothetical proteins (12). Using cell extracts of Escherichia coli expressing MM1085 fused to the maltose binding protein (MBP-NrpRI) and employing promoter binding studies by DNA-affinity chromatography demonstrated that MBP-NrpRI binds specifically to the nifH promoter. Deletion of various bases in the promoter region of nifH confirmed that the regulatory element ACC-N(7)-GGT is required for specific binding of NrpRI to the promoter. PMID- 18415080 TI - Introduction of a novel pathway for IAA biosynthesis to rhizobia alters vetch root nodule development. AB - We introduced into Rhizobium leguminosarum bv. viciae LPR1105 a new pathway for the biosynthesis of the auxin, indole-3-acetic acid (IAA), under the control of a stationary phase-activated promoter active both in free-living bacteria and bacteroids. The newly introduced genes are the iaaM gene from Pseudomonas savastanoi and the tms2 gene from Agrobacterium tumefaciens. Free-living bacteria harbouring the promoter-iaaMtms2 construct release into the growth medium 14-fold more IAA than the wild-type parental strain. This IAA overproducing R. l. viciae, the RD20 strain, elicits the development of vetch root nodules containing up to 60-fold more IAA than nodules infected by the wild-type strain LPR1105. Vetch root nodules derived from RD20 are fewer in number per plant, heavier in terms of dry weight and show an enlarged and more active meristem. A significant increase in acetylene reduction activity was measured in nodules elicited in vetch by RD20. PMID- 18415081 TI - The in vitro pharmacological profile of TD-5108, a selective 5-HT(4) receptor agonist with high intrinsic activity. AB - The in vitro pharmacological profile of TD-5108, a novel, selective 5-HT(4) receptor agonist, was compared to that of clinically efficacious gastroprokinetic 5-HT(4) receptor agonists. TD-5108 produced an elevation of cyclic adenosine monophosphate in human embryonic kidney 293 cells expressing the human recombinant 5-HT(4(c)) (h5-HT(4(c))) receptor (pEC(50) = 8.3) and 5-HT(4) receptor-mediated relaxation of the rat esophagus (pEC(50) = 7.9) and contraction of the guinea pig colon (pEC(50) = 7.9). In all in vitro assays, TD-5108 was a high intrinsic activity agonist, unlike tegaserod, mosapride, and cisapride which, in the majority of test systems, had lower intrinsic activity. TD-5108 had high affinity (pK (i) = 7.7) and selectivity (> or =25-fold) for h5-HT(4(c)) receptors over other biogenic amine receptors. TD-5108 was >500-fold selective over other 5-HT receptors (including h5-HT(2B) and h5-HT(3A)) and, at 3 microM, had no effect on human ether-a-go-go-related gene K+ channels. In conclusion, TD 5108 is a selective 5-HT(4) receptor agonist in vitro. The high intrinsic activity and preferential binding of TD-5108 to 5-HT4 over other 5-HT receptors may result in an improved clinical profile for the treatment of gastrointestinal disorders of reduced motility. PMID- 18415083 TI - Electrode modified with toluidine blue-doped silica nanoparticles, and its use for enhanced amperometric sensing of hemoglobin. AB - Three-dimensionally structured, silica based, organic-inorganic hybrid nanoparticles (NPs) were prepared by a simple and feasible water-in-oil (W/O) microemulsion method and a promising platform for bioelectrochemical analysis was obtained. The commonly used phenathiazine organic compound, toluidine blue (TB) was readily captured in the three-dimensional cage of the inorganic SiO(2) network, which was considered to serve as a protective "shell" toward the embedded TB. A TEM image indicated the size of the thus prepared TB-doped SiO(2) (TB@SiO(2)) NPs was 21 +/- 3 nm. UV-visible and IR spectroscopy confirmed successful formation of the organic-inorganic composite and possible interaction between TB and SiO(2), which favored enhanced stability of the hybrid. A sensitive amperometric sensor for hemoglobin (Hb) biomolecules based on TB@SiO(2) NPs conjugated with a biopolymer chitosan (CHIT) membrane was then developed. The surface of the silica NPs was highly biocompatible and the TB captured inside maintained its high electron-transfer efficiency. Dye leakage of TB from the TB@SiO(2) hybrid was proved to be minimal, owing to the inorganic SiO(2) network and the force of interaction between TB and SiO(2). The amperometric sensor had a detection limit of 2.5 x 10(-9) mol L(-1) (S/N = 3) with a linear range from 5.0 x 10(-9) to 3.0 x 10(-6) mol L(-1) for Hb. When it was applied to determine the concentration of a clinical blood sample, satisfactory results were obtained which were in good agreement with those obtained by the standard method. PMID- 18415082 TI - Repeated social defeat stress-induced sensitization to the locomotor activating effects of d-amphetamine: role of individual differences. AB - RATIONALE: In this study, we sought to examine individual differences in stress induced behavioral sensitization to d-amphetamine after repeated social defeat stress. In an effort to understand what mechanisms underlie stress-induced sensitization to d-amphetamine, we examined striatal gene expression of the dopamine receptor D(2). Additionally, we investigated if repeated social defeat was associated with changes in dendritic spine density in the hippocampus, prefrontal cortex, and nucleus accumbens of rats that exhibit stress-induced sensitization. METHODS: Male rats were classified into high responders (HR) and low responders (LR) based on their locomotor response to a novel environment. Then, rats were either handled as a control or defeated on four occasions by aggressive rats. Two weeks after the last defeat, animals were challenged with one of three doses of d-amphetamine and their locomotor activity was recorded. RESULTS: Non-defeated HR rats exhibited higher locomotor activity in response to d-amphetamine when compared to LR non-defeated rats. Fourteen days from the last repeated social defeat, LR rats and HR rats were behaviorally identical in response to acute injections of amphetamine. Furthermore, HR non-defeated rats had less D(2) mRNA expression in the nucleus accumbens core and dorsal striatum than do LR non-defeated rats. However, after repeated social defeat, HR and LR rats had identical D(2) mRNA expression in both the core and dorsal striatum. Finally, there were no changes in dendritic spine density in any of the brain areas examined in LR rats. CONCLUSION: Repeated social defeat abolishes individual differences in behavioral responses to d-amphetamine which may be due to a down-regulation of striatal dopamine D(2) receptors in LR rats. PMID- 18415084 TI - Single-drop microextraction and gas chromatography-mass spectrometry for the determination of volatile aldehydes in fresh cucumbers. AB - Headspace single-drop microextraction (HS-SDME) was used as a rapid and reliable method for the isolation and preconcentration of volatile aldehydes from fresh cucumbers. The utility of this methodology is demonstrated in the determination of (E)-2-nonenal and (E,Z)-2,6-nonadienal. The limit of detection, linearity and repeatability have been determined for 2,6-nonadienal and (E)-2-nonenal. Limits of detection for nonenal and nonadienal were 0.05 and 0.04 mg kg(-1), respectively. The repeatability of extraction was obtained with the RSD values lower than 13%. Concentrations of target aldehydes in fresh cucumbers obtained by means of the HS-SDME method were in the range 9.4-12.5 (nonadienal) and 2.6-3.8 mg kg(-1) (nonenal). The results of the single-drop extraction in combination with gas chromatography show promising potential for the analysis of volatile aldehydes in vegetables. PMID- 18415085 TI - Analysis of carbamate pesticides in water samples using single-drop microextraction and gas chromatography-mass spectrometry. AB - Single-drop microextraction (SDME) followed by gas chromatography-mass spectrometry detection was used for the determination of some carbamate pesticides in water samples. The studied pesticides were thiofanox, carbofuran, pirimicarb, methiocarb, carbaryl, propoxur, desmedipham and phenmedipham. Two alternative sample introduction methods have been examined and compared; SDME followed by cool on-column injection (without derivatization) and SDME followed by in-microvial derivatization and splitless injection. Acetic anhydride was used as derivatization reagent. Parameters that affect the derivatization reaction yield and the extraction efficiency of the SDME method were studied and optimized. The analytical performances and possible applications of both approaches were investigated. Relative standard deviations for the studied compounds ranged from 3.2 to 8.3%. The detection limits obtained by the derivatization method were found to be in the range 3-35 ng/L. Using cool on column injection (without derivatization), the detection limits were between 30 and 80 ng/L. PMID- 18415086 TI - Determination of boron in water samples at nanograms per cubic decimeter levels by reversed-phase partition high-performance liquid chromatography with precolumn complexation reaction using salicylaldehyde and 1-amino-8-naphthol-3,6 disulfonate. AB - A new method for the highly sensitive and selective determination of boron at nanograms per cubic decimeter levels has been developed based on the derivatization reaction of boron using salicylaldehyde and 1-amino-8-naphtol-3,6 disulfonate with reversed-phase partition high-performance liquid chromatography. A detection limit as low as 2.0 nmol/dm(3) (22 ng/dm(3)) was achieved without any preconcentration. No significant interference was observed in the determination of 16 micromol/dm(3) of boron with the addition of nine metal ions (Al(III), Cu(II), Co(II), Fe(II), Fe(III), Ni(II), Mn(II), V(V), Zn(II)) at concentrations 100 times greater than that of boron without any masking procedure. The proposed method was successfully applied to the determination of boron in river water, tap water, doubly distilled water, and highly purified water. PMID- 18415088 TI - Determination of trace elements in bone by two-jet plasma atomic emission spectrometry. AB - This paper describes an analytical method for trace element determination in bone tissues. The study of the influence of the bone matrix showed that the addition of 25% ground bone to graphite powder with introduced impurities did not affect the analytical signal of elements in the spectral excitation in a two-jet plasma. On basis of these investigations a method for direct multielement analysis of bone tissues was suggested. The sample preparation procedure consisted in mixing powdered bone (particle size 30 MUm or less) with a spectroscopic buffer (graphite powder plus NaCl) in ratio 1:3 or to a greater extent depending on the analyte concentration. Reference samples based on graphite powder were used for construction of calibration curves. The NaCl concentration in analyzed and calibration samples was 15 wt%. The effect of particle size was revealed from the determination of Ba, Sr, and Mg. To eliminate this effect, treatment of the samples with nitric acid was proposed. The validation of the technique was confirmed by comparison of the analysis results of a bone sample with those obtained by inductively coupled plasma atomic emission spectrometry after wet acid digestion. The limits of detection estimated for 20 elements were the following (MUg g(-1)): 0.1 (Ag), 1.0 (Al), 1.0 (Ba), 0.1 (Be), 1.2 (Bi), 0.4 (Cd), 1.0 (Co), 0.2 (Cu), 0.6 (Cr), 1.9 (In), 2 (Fe), 0.3 (Ga), 0.4 (Mn), 0.4 (Mo), 0.7 (Ni), 1.0 (Pb), 0.7 (Sn), 0.8 (Tl), 5 (Sr), 1.0 (Zn). PMID- 18415087 TI - Highly efficient analysis of underivatized carbohydrates using monolithic-silica based capillary hydrophilic interaction (HILIC) HPLC. AB - A polyacrylamide (PAAm)-modified monolithic silica capillary column of increased phase ratio, 200T-PAAm, for hydrophilic interaction liquid chromatography (HILIC) was prepared. The column showed high separation efficiency, with a theoretical plate height H = 7-20 microm at a linear velocity, u = 1-7 mm/s. From a kinetic plot analysis, it was expected that the monolithic column could provide three times faster separation than particle-packed HILIC columns under a pressure limit at 20 MPa. HILIC coupled with electrospray ionization (ESI)-mass spectrometry (HILIC-ESI-MS) using the 200T-PAAm column was employed for the analysis of underivatized carbohydrates to achieve fast and efficient separations of mixtures containing mono-, di-, and trisaccharides within 5 min. Under single MS full scan mode, 200 pg of oligosaccharides was detected by the system. The limit of detection (LOD) of the LC-ESI-MS/MS system was determined using selected reaction monitoring (SRM) to be as low as 3.2 ng/mL (attomol level) for nonreducing saccharides. The system was successfully applied to the detection of disaccharides in extracts of plant, such as corn, soybean, and Arabidopsis thaliana. PMID- 18415089 TI - Production of malachite green oxalate and leucomalachite green reference materials certified for purity. AB - Malachite green oxalate (MG oxalate) and leucomalachite green (LMG) have been prepared and certified as pure reference materials. The purities of MG oxalate and LMG were assessed by high-performance liquid chromatography-diode array detection (HPLC-DAD), nuclear magnetic resonance (NMR) spectroscopy, differential scanning calorimetry (DSC), Karl Fischer titration, ashing and thermogravimetric analysis (TGA). MG oxalate was purified by supercritical fluid extraction (SFE). Prior to purification, commercial MG oxalate purity was estimated to be about 90%. The main impurities present in SFE-purified MG oxalate were identified and quantified using HPLC-DAD. The main impurities were found to be monode-MG (monodemethylated MG oxalate synthesis impurity), 4-(dimethylamino)benzophenone (4-DMABP), MG-carbinol and LMG. The homogeneity of both reference materials was also determined. Issues associated with the stability of LMG and MG oxalate in solution forced an extensive study investigating different parameters i.e. solvent, acid, analyte concentration and temperature. MG oxalate (100 microg/mL) was found to be stable in acetonitrile containing 1% v/v glacial acetic acid for at least 155 days and LMG (100 microg/mL) was stable in acetonitrile for at least 133 days. The final purity value for MG oxalate was 94.3 +/- 1.4% m/m at the 95% confidence interval (or 67% m/m if MG cation is reported). For LMG, the certified purity was found to be 98.8 +/- 0.8% m/m at the 95% confidence interval. PMID- 18415090 TI - Simultaneous determination of six major ergot alkaloids and their epimers in cereals and foodstuffs by LC-MS-MS. AB - This paper describes a new and rapid method for accurate quantification of the six ergot alkaloids, ergometrine, ergotamine, ergosine, ergocristine, ergocryptine, and ergocornine, by liquid chromatography-tandem mass spectrometry (LC-MS-MS). The six ergot alkaloids studied have been defined by the European Food Safety Authority (EFSA) as among the most common and physiologically active ones. In addition, the method enables the quantification of the corresponding six epimers (ergo-inines) of these ergot alkaloids. This is of considerable importance in terms of the differences in toxicity of the isomeric forms. The method involves extraction under alkaline conditions using a mixture of acetonitrile and ammonium carbonate buffer followed by a rapid clean-up using dispersive solid-phase extraction with PSA (primary secondary amine) and a short chromatographic LC-run (21 min) with subsequent MS-MS detection. The method was developed and validated using ten different cereal and food samples. The major strength of the new method compared with previously published techniques is the simplicity of the clean-up procedure and the short analysis time. The limits of quantification were 0.17 to 2.78 MUg kg(-1) depending on the analyte and matrix. Recovery values for the 12 ergot alkaloids spiked into ten different matrices at levels of 5, 50, and 100 MUg kg(-1) were between 69 and 105% for 85 of 90 recovery measurements made over six days. Measurement uncertainty values were highly satisfactory. At a concentration level of 5 MUg kg(-1) the expanded measurement uncertainty ranged from +/-0.56 to +/-1.49 MUg kg(-1), at a concentration level of 100 MUg kg(-1) the expanded measurement uncertainty ranged from +/-8.9 to +/-20 MUg kg(-1). Both LOQs and measurement uncertainties were dependent on the analyte but almost independent of the matrix. The method performance was satisfactory when tested in a mini-intercomparison study between three laboratories from three different countries. PMID- 18415091 TI - Mineral oil certified reference materials for the determination of polychlorinated biphenyls from the National Metrology Institute of Japan (NMIJ). AB - Four mineral oil certified reference materials (CRMs), NMIJ CRM 7902-a, CRM 7903 a, CRM 7904-a, and CRM 7905-a, have been issued by the National Metrology Institute of Japan, which is part of the National Institute of Advanced Industrial Science and Technology (NMIJ/AIST), for the determination of polychlorinated biphenyls (PCBs). The raw materials for the CRMs were an insulation oil (CRM 7902-a and CRM 7903-a) and a fuel oil (CRM7904-a and CRM 7905 a). A solution of PCB3, PCB8, and technical PCB products, comprising four types of Kaneclor, was added to the oil matrices. The total PCB concentrations in the PCB-fortified oils (CRM 7902-a and CRM 7904-a) are approximately 6 mg kg(-1). In addition, the mineral oils which were not fortified with PCBs were also distributed as CRMs (CRM 7903-a and CRM 7905-a). Characterization of these CRMs was conducted by the NMIJ/AIST, where the mineral oils and the PCB solution were analyzed using multiple analytical methods such as dimethylsulfoxide extraction, normal-phase liquid chromatography, gel permeation chromatography, reversed-phase liquid chromatography, and chromatography using sulfoxide-bonded silica; and/or various capillary columns for gas chromatography, and two ionization modes for mass spectrometry. The target compounds in the mineral oils and those in the PCB solution were determined by one of the primary methods of measurement, isotope dilution-mass spectrometry (ID-MS). Certified values have been provided for 11 PCB congeners (PCB3, 8, 28, 52, 101, 118, 138, 153, 180, 194, and 206) in the CRMs. These CRMs have information values for PCB homologue concentrations determined by using a Japanese official method for determination of PCBs in wastes and densities determined with an oscillational density meter. Because oil samples having arbitrary PCB concentrations between respective property values of the PCB-fortified and nonfortified CRMs can be prepared by gravimetric mixing of the CRM pairs, these CRMs can be used for validation of PCB analyses using various instruments which have different sensitivities. PMID- 18415092 TI - The time course of amplitude specification in brief interceptive actions. AB - The interception of fast moving objects typically allows the object to be seen for only a short period of time. This limits the time available to prepare the movement. To deal with short preparation intervals, performers are likely to prepare a motor program in advance. Although motor preparation may begin before the target is seen, accuracy requires that certain program parameters are determined from observations of the target. In the experiments reported here we sought to determine the last moment at which information about the distance to move (amplitude) can be incorporated into a program. We employed an empirical protocol that allowed us to examine whether new amplitude information is incorporated discretely or continuously into the program during short intervals prior to movement onset (MO)-the preparation interval. Participants were trained to hit targets at two different distances with movements of a specific duration (180 ms): targets were moving in "Experiment 1" and stationary in "Experiment 2". This method permitted an estimate of MO time. Preparation intervals were manipulated by delivering a stimulus cue for movement amplitude at varying times prior to the estimated MO. Results demonstrated that amplitude information could be effectively incorporated into the program provided the preparation interval was greater than about 200 ms. In addition, the results indicated that amplitude was specified predominantly in a discrete manner, though the number of responses directed towards a central default amplitude suggest that the distance between targets was near to a threshold for continuous specification. PMID- 18415093 TI - Pharmacokinetics and tolerability of artesunate and amodiaquine alone and in combination in healthy volunteers. AB - OBJECTIVES: The WHO recommends artemisinin-based combination therapies for treatment of uncomplicated falciparum malaria. At least 15 African countries have adopted artesunate plus amodiaquine as treatment policy. As no pharmacokinetic data on this combination have been published to date, we investigated its pharmacokinetic interactions and tolerability in healthy volunteers in Africa. METHODS: In a randomized, three-phase, cross-over study, amodiaquine (10 mg/kg) and artesunate (4 mg/kg) were given as single oral doses to 15 healthy volunteers. Artesunate was given to all volunteers on day 0. On day 7 they received either amodiaquine or amodiaquine plus artesunate and the alternative regimen on day 28. The pharmacokinetics of artesunate and amodiaquine and their main active metabolites dihydroartemisinin and desethylamodiaquine were compared following monotherapy and combination therapy using analysis of variance. RESULTS: Thirteen volunteers completed the study, and pharmacokinetic parameters could be determined for twelve volunteers. When given in combination, the mean AUC was lower for dihydroartemisinin [ratio 67% (95% CI 51-88%); P = 0.008] and desethylamodiaquine [ratio 65% (95% CI 46-90%); P = 0.015] when compared with monotherapy. Adverse events of concern occurred in four volunteers (27%): grade 3 transaminitis (n = 1), neutropaenia (n = 2), and hypersensitivity (n = 1). CONCLUSION: The total drug exposure to both drugs was reduced significantly when they were given in combination. The clinical significance of these interactions is unclear and must be studied in malaria patients. The frequency and nature of adverse events among the healthy volunteers were of concern, and suggest laboratory monitoring would be needed in malaria patients treated with artesunate plus amodiaquine. PMID- 18415094 TI - Functional diversification of the toll-like receptor gene family. AB - Phylogenetic analyses supported the hypothesis that the vertebrate toll-like receptors (TLRs) include two very ancient groups that arose by gene duplication prior to the divergence of protostomes and deuterostomes: (1) the TLR1 family (including mammalian TLR1, TLR2, TLR6, and TLR10); and (2) a clade including the remainder of mammalian TLRs. Correlating data on ligand type, subcellular localization, and gene expression in leukocytes and other tissues with the phylogeny provided evidence that certain major functional specializations within the TLRs occurred after ancient gene duplication events and that these traits have been retained through further events of gene duplication. For example, the recognition of bacterial lipoproteins appears to have arisen in the ancestor of the TLR1 family and continues to characterize members of that family whose ligands are known. Likewise, expression on the endosomal membrane and the recognition of nucleic acids appears to have been arisen in the ancestor of the TLR7 family and some related TLRs. On the other hand, gene expression patterns across tissues appear to have been much more volatile over the evolution of the vertebrate TLRs, since genes may show expression profiles similar to those of distantly related genes but dissimilar to those of closely related genes. Thus, the vertebrate TLRs provide an example of a multi-gene family in which gene duplication has been followed by extensive changes in certain aspects of gene function, while others have been conserved throughout vertebrate history. PMID- 18415095 TI - Bifunctional enhancement of a beta-glucanase-xylanase fusion enzyme by optimization of peptide linkers. AB - The flexible peptides (GGGGS)n (n < or = 3), the alpha-helical peptides (EAAAK)n (n < or = 3) and two other peptides were used as linkers to construct bifunctional fusions of beta-glucanase (Glu) and xylanase (Xyl) for improved catalytic efficiencies of both moieties. Eight Glu-Xyl fusion enzymes constructed with different linkers were all expressed as the proteins of ca. 46 kDa in Escherichia coli BL21 and displayed the activities of both beta-glucanase and xylanase. Compared to all the characterized fusions with the parental enzymes, the catalytic efficiencies of the Glu and Xyl moieties were equivalent to 304 426% and 82-143% of the parental ones, respectively. The peptide linker (GGGGS)(2) resulted in the best fusion, whose catalytic efficiency had a net increase of 326% for the Glu and of 43% for the Xyl. The two moieties of a fusion with the linker (EAAAK)(3) also showed net increases of 262 and 31% in catalytic efficiency. Our results highlight, for the first time, the enhanced bifunctional activities of the Glu-Xyl fusion enzyme by optimizing the peptide linkers to separate the two moieties at a reasonable distance for beneficial interaction. PMID- 18415096 TI - Cloning of a rumen fungal xylanase gene and purification of the recombinant enzyme via artificial oil bodies. AB - A gene encoding a xylanase, named xynS20, was cloned from the ruminal fungus Neocallimastix patriciarum. The DNA sequence of xynS20 revealed that the gene was 1,008 bp in size and encoded amino acid sequences with a predicted molecular weight of 36 kDa. The amino acid sequence alignment showed that the highest sequence identity (28.4%) is with insect gut xylanase XYL6805. According to the sequence-based classification, a putative conserved domain of glycosyl hydrolase family 11 was detected at the N-terminus of XynS20 and a putative conserved domain of family 1 carbohydrate-binding module (CBM) was observed at the C terminus of XynS20. An Asn-rich linker sequence was found between the N-terminal catalytic domain and the C-terminal CBM of XynS20. To examine the activity of the gene product, xynS20 gene was cloned as an oleosin-fused protein, expressed in Escherichia coli, affinity-purified by formation of artificial oil bodies, released from oleosin by intein-mediated peptide cleavage, and finally harvested by concentration of the supernatant. The specific activity of purified XynS20 toward oat spelt xylan was 1,982.8 U mg(-1). The recombinant XynS20 was stable in the mild acid pH range from 5.0 to 6.0, and the optimum pH was 6.0. The optimal reaction temperature of XynS20 was 45 degrees C; at temperatures below 30 and above 55 degrees C, enzyme activity was less than 50% of that at the optimal temperature. PMID- 18415097 TI - RhoC a new target for therapeutic vaccination against metastatic cancer. AB - Most cancer deaths are due to the development of metastases. Increased expression of RhoC is linked to enhanced metastatic potential in multiple cancers. Consequently, the RhoC protein is an attractive target for drug design. The clinical application of immunotherapy against cancer is rapidly moving forward in multiple areas, including the adoptive transfer of anti-tumor-reactive T cells and the use of "therapeutic" vaccines. The over-expression of RhoC in cancer and the fact that immune escape by down regulation or loss of expression of this protein would reduce the morbidity and mortality of cancer makes RhoC a very attractive target for anti-cancer immunotherapy. Herein, we describe an HLA-A3 restricted epitope from RhoC, which is recognized by cytotoxic T cells. Moreover, RhoC-specific T cells show cytotoxic potential against HLA-matched cancer cells of different origin. Thus, RhoC may serve as an important and widely applicable target for anti-cancer immunotherapeutic strategies. PMID- 18415098 TI - Tibial lengthening using a reamed type intramedullary nail and an Ilizarov external fixator. AB - The aim of this study was to evaluate the efficacy of tibial lengthening using a reamed type intramedullary nail and an Ilizarov external fixator for the treatment of leg length discrepancy or short stature. This retrospective study was performed on 18 tibiae (13 patients) in which attempts were made to reduce complications. We used an Ilizarov external fixator and a nail (10 mm diameter in 17 tibiae and 11 mm in one tibia) in combination. Average limb lengthening was 4.19 cm (range, 2.5-5.5). The mean duration of external fixation was 12.58 days per centimetre gain in length, and the mean consolidation index was 40.53 (range, 35.45-51.85). All distracted segments healed spontaneously without refracture or malalignment. Gradual limb lengthening using a reamed type intramedullary nail and circular external fixation in combination was found to be reliable and effective and reduced external fixation time with fewer complications. PMID- 18415099 TI - The effect of porosity and mechanical property of a synthetic polymer scaffold on repair of osteochondral defects. AB - We have made three types of poly (DL-lactide-co-glycolide) (PLG) scaffolds (porosity: scaffold I 80 +/- 0.9%, II 85 +/- 0.8%, III 92 +/- 0.7%; compression module determined with 10% strain: scaffold I 0.26 MPa, II 0.091 MPa, III 0.0047 MPa). Osteochondral defects made in the femoral condyle of rabbits were treated with these scaffolds and the possibilities of cartilage repair were investigated histologically. At post-operative weeks 6 and 12, histological scores in the groups of scaffolds II and III were significantly higher than the score in the group of scaffold I. Scaffolds II and III, which have higher porosity than scaffold I, allow better migration of bone marrow cells and better replacement of the scaffold with bone and cartilage than scaffold I. This study suggests that higher porosity allowing bone marrow cells to migrate to the scaffold is important in repairing osteochondral defects. PMID- 18415100 TI - Phase II trial of biweekly paclitaxel plus infusional 5-fluorouracil and leucovorin in patients with advanced or recurrent inoperable gastric cancer. AB - PURPOSE: To investigate the efficacy and safety of combination chemotherapy with biweekly paclitaxel plus infusional 5-fluorouracil and leucovorin in the treatment of patients with advanced or metastatic gastric cancer. PATIENTS AND METHODS: Chemonaive patients with histologically confirmed advanced or recurrent inoperable gastric cancer were enrolled in the present study. Treatment consisted of paclitaxel (75 mg/m(2)) and leucovorin (40 mg/m(2)) as a 2-h intravenous infusion, followed by 5-fluorouracil (2,400 mg/m(2)) as a 46-h continuous infusion. Cycles were repeated every 2 weeks. RESULTS: Thirty patients were enrolled in this study. There were 12 partial responses, giving an overall response rate of 40.0%. At a median follow-up of 10.6 months, the median time to progression and median overall survival were 3.9 and 8.8 months, respectively. The most common hematological toxicity was grade 1-2 anemia, which was seen in 83.3% of patients. No grade 4 leukopenia, thrombocytopenia, or anemia was noted. The most common non-hematological toxicity was anorexia, which was seen in 70% of patients, although grade 3 anorexia was noted in only 10% of cases. There was no severe treatment-related morbidity or death. CONCLUSION: Combination chemotherapy consisting of biweekly paclitaxel plus infusional 5-fluorouracil and leucovorin was effective and well tolerated in patients with advanced gastric cancers. PMID- 18415101 TI - Functional analysis of a pollen-expressed polygalacturonase gene BcMF6 in Chinese cabbage (Brassica campestris L. ssp. chinensis Makino). AB - In our earlier work, a pollen-expressed polygalacturonase gene BcMF6 was isolated from floral bud of Chinese cabbage (Brassica campestris L. ssp. chinensis Makino) by cDNA-amplified fragment length polymorphism (cDNA-AFLP) transcript profiling and rapid amplification of cDNA ends (RACE). To unravel the biological function of BcMF6 gene, the antisense fragment from the BcMF6 gene with A9 promoter and CaMV35S promoter was transferred into flowering Chinese cabbage (B. campestris ssp. chinensis var. parachinensis). Out of transgenic plants transformed with the antisense gene constructed from the BcMF6, transgenic line with A9 promoter have a similar appearance to that with CaMV35S promoter. Morphological investigations showed that the transgenic plants developed the smaller floral organ with thin anther and less pollen. Pollen germination test indicated that only near 50% the pollen from the transgenic line could normally germinate. Further scanning electron microspore analysis of transgenic plants confirmed that half of pollen was abnormal. Cytological comparisons of microspore development also demonstrated that process of microsporogenesis was held up, microspores maturation was disrupted and pollen grain fail to separate, finally. In a word, the present study revealed that BcMF6, as a polygalacturonase gene, has a role in pollen maturation and pollen tube growth. PMID- 18415102 TI - [Fundus autofluorescence in patients with inherited retinal diseases : patterns of fluorescence at two different wavelengths]. AB - BACKGROUND: Fundus autofluorescence (FAF) may be excited and measured at different wavelengths. In the present study we compared short wavelength and near infrared FAF patterns of retinal dystrophies. METHODS: We analysed both eyes of 108 patients with diverse retinal dystrophies. Besides colour fundus photographs, FAF images were obtained with the Heidelberg Retina Angiograph (HRA 2). Excitation wavelengths of 488 nm (blue; filter at 500 nm) and 787 nm (near infrared; filter at 810 nm) were applied. For improvement of the signal-to-noise ratio a total of nine images were averaged, and the mean images (original grey values, not normalized) were analysed. RESULTS: Useful FAF images of both excitation wavelengths were achieved in all patients. We observed characteristic FAF patterns, which differed between excitation wavelengths depending on the disease. With time, FAF pattern changes and progression could be observed. CONCLUSION: FAF of both wavelengths provided additional information for phenotype description in retinal dystrophies. Other than short wavelength FAF, near infrared FAF showed different pathological changes, which may be related to changes in RPE melanin. However, any conclusions may be limited by the still incomplete knowledge about the prognostic value of FAF in the diseases studied here. PMID- 18415103 TI - [The aspheric blue light filter IOL AcrySof IQ compared to the AcrySof SA60AT : influence of IOL power, pupil diameter, and corneal asphericity on postoperative spherical aberration]. AB - BACKGROUND: The aspheric optic design of intraocular lenses (IOL) aims to minimize postoperative ocular spherical aberration (SA). The effect of a standardized IOL asphericity depends on IOL power, pupil diameter, and corneal asphericity. The impact of these factors was investigated in a comparative study. MATERIAL AND METHODS: In a prospective study, 70 eyes with the aspheric blue light filter IOL AcrySof IQ (Alcon) were compared to 36 eyes with the conventional AcrySof SA60AT (Alcon). Six weeks after uneventful phacoemulsification the following parameters were assessed: visual acuity, higher order aberrations, pupil diameter, and corneal asphericity. RESULTS: The comparison of 42 eyes in a normal dioptric range with the IQ IOL (22.4+/-2.0 D) to 20 eyes with the control lens of similar IOL power (22.0+/-2.7 D) showed no different visual outcome. For a 5-mm pupil SA was calculated at 0.04 (+/-0.05) microm for the IQ IOL and 0.20 (+/-0.06) microm for the AcrySof SA60AT. Also for a 4-mm pupil SA was significantly lower in eyes with the IQ lens. A separate evaluation of 28 eyes which needed very high or low IOL power revealed a similar postoperative SA for the IQ IOL. The greatest difference of SA between the two IOLs was found in hyperopic eyes with higher IOL power. The corneal asphericity (Q value) showed significant correlation to postoperative ocular SA. CONCLUSIONS: The aspheric AcrySof IQ reduces postoperative SA significantly compared to a conventional IOL even with smaller pupils and independent of IOL power. The assessment of corneal asphericity enables an individual estimation of postoperative SA. PMID- 18415104 TI - [Measurement of viscoelastic corneal parameters (corneal hysteresis) in patients with primary open angle glaucoma]. AB - BACKGROUND: The ocular response analyzer (ORA) uses an air-pressure-triggered, dynamic, bi-directional corneal applanation method to measure biomechanical parameters of the cornea. Corneal hysteresis (CH) is defined as the difference in intraocular pressure recorded during inward and outward applanation. CH is therefore an indicator for the viscoelastic properties of the cornea. PATIENTS AND METHODS: CH was recorded in non-glaucoma patients (80 eyes) as well as in patients with primary open angle glaucoma (POAG, 82 eyes). The correlation between CH and central corneal thickness (CCT) was analyzed. RESULTS: Mean CH was 10.6+/-2.2 mmHg in the non-glaucoma group and 9.3+/-2.2 mmHg in patients with POAG (p<0.01). CH and CCT showed a positive correlation in non-POAG patients, however no such correlation was found in the POAG group. CONCLUSION: Patients with POAG show an alteration of biomechanical corneal parameters with a significant decrease in corneal hysteresis. A positive correlation between CH and CCT, which was seen in the non-glaucoma group could not be detected in the POAG group. PMID- 18415105 TI - [Sudden loss of vision]. PMID- 18415107 TI - [Tamoxifen retinopathy: a case series of clinical and functional data]. AB - BACKGROUND: Tamoxifen is used in the treatment of selected patients with breast carcinoma. Rarely, it has been shown to cause ocular toxic effects including crystalline retinopathy. METHODS: Retrospective analysis of clinical and functional (visual acuity, visual field, colour vision) data of a case series of eight female patients under tamoxifen therapy with electrophysiological examination. RESULTS: Seven of eight patients complained of visual disturbances. In one case, examination showed crystalline deposits in the cornea and macular area. Three patients revealed changes in full-field and multifocal electroretinogram, and two had a pathological multifocal electroretinogram only. In six cases we applied a desaturated panel D-15 colour vision test; five of these showed some disorders. CONCLUSIONS: Most tamoxifen patients who complained of visual disturbances showed electrophysiological changes, particularly in the multifocal electroretinogram and often without a certain morphological correlate. We recommend electrophysiological examination for patients with unclear visual deterioration who are receiving tamoxifen therapy. PMID- 18415108 TI - [Peripheral suprathreshold stimulation in preperimetric glaucoma]. AB - BACKGROUND AND PURPOSE: Early glaucomatous visual field defects can occur outside the central 30 degrees , which is usually examined in perimetric tests used for glaucoma diagnosis and screening. This study aimed to evaluate the diagnostic value of peripheral suprathreshold stimulation in open angle glaucoma before the development of reproducible visual field damage in standard 30 degrees automatic white-on-white perimetry. METHODS: A total of 352 eyes of 352 patients (ages 35 69 years; visual acuity 0.8 or better) from the Erlanger Glaucoma Registry were included in this study. They were divided into two groups: normal eyes and preperimetric glaucoma. All patients underwent a standardized glaucoma examination including Octopus 500EZ static perimetry (G1 program, all three phases); 95 eyes of 95 patients also received a 135-point suprathreshold test pattern of the Humphrey Field Analyzer (model 750i) for detecting peripheral visual field defects. Sensitivity and specificity were calculated for any single test point in phase 3 of the G1 test pattern and the Humphrey 135-point pattern. A score was calculated, and cluster analysis was performed. RESULTS: In 33 of 176 (18.8%) eyes with preperimetric glaucoma, the score was 3 or higher in phase 3 of the G1 program (normal eyes: 19 of 196; 9.7%). For both examination modalities, the highest sensitivity was found in test locations in the superior nasal midperiphery, corresponding to neuroretinal rim loss predominantly in the inferotemporal sector in early glaucomatous optic disc atrophy. CONCLUSION: Positive test results using suprathreshold stimulation in the midperiphery can be found in patients with preperimetric glaucoma at a significantly higher frequency than in normal subjects. Longitudinal studies will show whether such tests can be useful for predicting perimetric manifestation of the disease. PMID- 18415109 TI - [A rare orbital tumor]. PMID- 18415111 TI - Elbow arthrolysis in severely stiff elbows. AB - INTRODUCTION: This study involves the results of open elbow arthrolysis performed on a series of patients having fixed joint contracture. PATIENTS AND METHODS: Eighteen patients were treated with open arthrolysis. Eleven patients had very severely stiff elbows (flexion arc less than 30 degrees ), five patients had severely stiff elbows (31 degrees -60 degrees ) and two patients had moderately stiff elbows (60 degrees -90 degrees ). Fixed flexion deformity was observed in nine patients preoperatively.The mean interval between the trauma and arthrolysis was 14.8 months. The mean age was 27 years. After radiological examination, lateral kocher incision was used on 13 patients, medial and lateral incision on 3 patients, transolecranon approach on 1 patient and anterior and posterior approach on 1 patient. External fixator was applied on four patients. Fasia lata interposition was performed in three patients. Additional procedures were as follows, bone fixation in five patients, bone grafting in two patients, nerve grafting in one patient, subcutaneous ulnar nerve transposition in three patients. The average follow-up time was 47 months. RESULTS: At the final evaluation, the mean extension deficit had improved from 55 degrees to 32 degrees . The mean end flexion increased from 81 degrees to 124 degrees postoperatively. The flexion arc of three patients in whom heterotopic ossification was excised had increased to 65 degrees . Infection was diagnosed in two patients (11%). CONCLUSION: The importance of this study is that the patients have fixed deformities and a long follow-up time. Severely stiff elbow is one of the main indications of open arthrolysis in the patients without muscle atrophy. We suggest sequential arthrolysis as an effective way to obtain good range of motion especially in severe stiff elbows as well as to maintain the ligamantous stability of the elbow joint. PMID- 18415110 TI - A large discoidal lipoprotein present in only one of two closely related crayfish. AB - The hemolymph lipoproteins of two European freshwater crayfish, Astacus astacus and Astacus leptodactylus, were isolated and characterized. The former species possesses two sex-independent lipoproteins, which can be related to the formerly described high-density lipoprotein (HDL)/beta-glucan binding protein and very high-density lipoprotein/clotting protein from other crustaceans. The latter species, however, contains an additional third lipoprotein with a unique structure. It is a large discoidal HDL with a diameter of 42 nm, a thickness of 7 nm and a density of 1.1 g/ml. SDS-PAGE revealed two different apolipoproteins with molecular masses of 240 and 85 kDa, respectively, arranged in a 1:1 stoichiometry as judged from cross linking experiments. The lipid content of this lipoprotein was 67%, far higher than in every other crustacean lipoprotein described so far. The native molecular mass of this HDL-type lipoprotein was estimated to be about 930 kDa. The lipid content of the other lipoproteins ranged between 25 and 30% for the HDL/beta-glucan binding protein and 6-8% for the VHDL/clotting protein. PMID- 18415112 TI - The use of structural periacetabular allografts in acetabular revision surgery: 2.5-5 years follow-up. AB - INTRODUCTION: "Acetabular bone loss" presents a major reconstructive challenge in total hip arthroplasty. Loss of acetabular bone stock is a consequence of removal of bone during the original procedure, subsequent prosthetic failure and osteolysis resulting from wear particles of cement and polyethylene. In case of severe bone loss treatment options are rather limited, as fixation requires either biological (cancellous allograft with cage or structural allograft) or non biological (trabecular metal, triflange implant, etc.) scaffolds. MATERIALS AND METHODS: Fourteen acetabular revisions with a cemented cup, supported by a deep frozen structural periacetabular allograft without using a reinforcement ring or an antiprotrusio cage were performed. Clinical assessment was done using a Harris hip score. Graft resorption was radiographically quantified using a digital measurement program. RESULTS: Kaplan-Meier survivorship was 67.1% at 42 months. After a mean follow-up of 3.6 years, 9 out of 14 patients had good clinical results. Mean resorption of 17.1% in six out of ten patients was observed. No sound evidence for union was found in nine out of ten patients. In the four re revised patients, no bone stock restoration was found. CONCLUSION: The bicortical allograft without protective device seems to function as a passive biocompatible dead scaffold, which has a less intrinsic strength at intermediate term follow up, compared to the by ring or cage protected structural allografts. PMID- 18415113 TI - A survey of satisfaction and use among patients fitted with a BAHA. AB - We compared the indices of satisfaction and use among patients wearing an osseo integrated prosthesis BAHA (bone anchored hearing aid) according to the indications: conductive or mixed hearing loss (CHL) and patients with single side deafness (SSD). The study was carried out among patients wearing a BAHA fitted in one of three French departments between November 2001 and November 2005. Each patient received a postal questionnaire relating to the ease of use and the daily utilization period of the prosthesis, as well as a satisfaction rating (from 1 to 10) evaluating improvement in quality of life, overall satisfaction, improvement in sound localization and satisfaction from the aesthetic point of view. In total, 170 out of 231 patients responded to the questionnaire (response rate of 73.5%). The average age at the time of fitting of the BAHA was 56 years (18-79 years). The SSD group was composed of 118 patients (69.4%): 92 following surgery for vestibular schwannoma, 2 following surgery for meningioma and 24 with "other" causes (e.g. idiopathic sudden deafness, sensori neural hearing loss complicating surgery of the middle ear). The CHL group was composed of 52 patients (30.5%): 44 patients with a chronic otitis and 8 with a malformation of the middle ear. The average duration of use of the prosthesis was 22 months (3-72 months). The average utilization period was higher than 8 h per day in 48.5% of cases. There was a significant difference between the two groups concerning the quality of life (p < 0.0001), general satisfaction (p < 0.0001) and sound localization (p < 0.01). There was no significant difference concerning aesthetics. Among the patients of the CHL group, the levels of satisfaction and quality of life are comparable with recent data in the literature with scores of good or very good. The BAHA thus remains one of the methods of choice for hearing rehabilitation in this group of patients. Among the patients of the SSD group, the levels of satisfaction and quality of life are significantly poorer than in the CHL group, but remain generally good with the exception of sound localization. The treatment of SSD patients with a BAHA is interesting. A study comparing the BAHA with the WIFI CROS system is justified in order to ascertain the respective advantages of these two treatment options. PMID- 18415114 TI - Superior semicircular canal dehiscence: positive predictive value of high resolution CT scanning. AB - Patients with superior dehiscence (SCD) syndrome present with vertigo and oscillopsia evoked by loud sounds and changes in middle ear or intracranial pressure. The first objective of this retrospective cohort study is to demonstrate that thin-section computed tomography (CT) scans reformatted in the plane of the superior semicircular canal (SSC) overestimate this anomaly compared to pathologic studies. The second objective of this study is to re-evaluate the positive predictive value of temporal bone scanning. All temporal bone CT scans with 0.55-mm collimation and reconstruction in the SSC plane performed over a 1 year period were analysed at a tertiary referral centre. CT-positive cases had their clinical data reviewed and patients were re-examined, if available. A total of 581 temporal bone CT-scans were analysed. A dehiscent-appearing superior canal was seen in 4.0% of studies while published pathologic studies report that only 0.5% of temporal bones SSCs have a dehiscence (P < 0.001). Of the 21 patients with positive temporal bone CTs, only 1 presented with sufficient clinical dues to identify the syndrome. Three additional patients did not have symptoms consistent with the diagnosis, but had surgery for a dehiscence of the tegmen mastoideum. When our findings are added to published data, the positive predictive value of temporal bone CT-scanning drops from 93 to 57%. The prevalence of dehiscent-appearing superior canal on thin-section temporal bone scanning with reformation in the SSC plane is much higher than anticipated by pathologic studies. Even with 0.55 mm-collimated helical CT and reformation in the SSC plane, the risk of overdiagnosis is present. PMID- 18415115 TI - Use of a single bipolar electrode in the posterior arytenoid muscles for bilateral monitoring of the recurrent laryngeal nerves in thyroid surgery. AB - The aims were to assess the technical feasibility of using a single electrode in the posterior arytenoid muscles (PAM) for intraoperative monitoring of the recurrent laryngeal nerve (RLN) in thyroid surgery, to validate the new method against the insertion of electrodes placed in the vocal cord muscle, and to report the results of the clinical application of the new concept. A total of 52 patients were enrolled. The handling and safety of RLN monitoring was tested by simultaneous registration of the EMG response from vocal fold electrodes and PAM electrodes. Acoustically and electromyographically we found nearly the same values for the arytenoid muscles as for the vocal folds, although the signals taken from the vocal folds were slightly stronger. PAM recording using a single bipolar electrode is technically feasible and as reliable compared to the standard vocal cord monitoring. PMID- 18415117 TI - Vigorous physical activity and cytokines in adolescents. AB - Habitual physical activity and chronic exercise may positively influence circulating cytokines in youth as in adults. The purpose of this study was to determine if adolescents reporting higher amounts of physical activity had a better cytokine profile independent of weight status. We compared the concentrations of cytokines in highly active (n = 60) and less active adolescents (n = 60) of different weight status [normal weight (BMI < 75th percentile), and overweight (BMI > 95th percentile)]. Vigorous physical activity (VPA) was obtained through a questionnaire and included activities with a MET level of > or =6. Adiponectin, resistin, TNF-alpha and IL-6 were measured in fasting plasma samples. Anovas indicated that girls with high VPA had higher adiponectin than girls with low VPA (P < 0.05), but similar resistin, TNF-alpha and IL-6 concentrations (P > 0.05). Boys with high VPA had only a trend to lower resistin than those with low VPA (P = 0.052). Differences between VPA groups were independent of weight status. In adolescents, higher amounts of accumulated habitual VPA may improve some cytokines, reducing the risk of inflammation associated with increased adiposity and insulin resistance. The effect of physical activity appears to be independent of weight status. PMID- 18415116 TI - Tight junctions and the modulation of barrier function in disease. AB - Tight junctions create a paracellular barrier in epithelial and endothelial cells protecting them from the external environment. Two different classes of integral membrane proteins constitute the tight junction strands in epithelial cells and endothelial cells, occludin and members of the claudin protein family. In addition, cytoplasmic scaffolding molecules associated with these junctions regulate diverse physiological processes like proliferation, cell polarity and regulated diffusion. In many diseases, disruption of this regulated barrier occurs. This review will briefly describe the molecular composition of the tight junctions and then present evidence of the link between tight junction dysfunction and disease. PMID- 18415118 TI - Neuromuscular fatigue is greater following highly variable versus constant intensity endurance cycling. AB - The present study compared neuromuscular fatigue of the knee extensor muscles following highly variable versus constant power output cycling. Ten subjects performed two 33-min cycling trials of the same average power output, in a random order. Cycling exercise was performed either at constant (CST) power output, corresponding to 70% of the maximal aerobic power (MAP), or at variable (VAR) power output with alternating high (200, 150 and 100% of MAP during 10, 15 and 20 s, respectively) and moderate (50% of MAP) power output periods. Neuromuscular tests were performed before and immediately after the two trials. Heart rate (HR) was measured during exercise and blood lactate concentration ([La]) at the end of both trials. Reductions in maximal voluntary contraction torque, voluntary activation level and peak doublet were significantly greater after VAR than after CST. HR and [La] were significantly higher during VAR than during CST. Cycling at a varying power output in comparison to constant power resulted in additional muscular fatigue that may be explained by greater anaerobic contribution and muscle solicitation during the highly variable power output protocol. PMID- 18415119 TI - Motivation, cohesion, satisfaction, and their relation to stress fracture among female military recruits. AB - The purpose of this study was to compare ratings of motivation, satisfaction, self-efficacy, and cohesion between male and female soldiers undergoing basic training (BT) in a gender-integrated unit and to study whether the ratings differ in a subgroup of women who suffered stress fractures (SF) during the course of training. Data were collected from 3 different companies of an integrated combat unit using questionnaires that were completed by 41 male and 160 female soldiers (age 18.5 +/- 0.4 years) at 3 phases during their BT (0, 2, and 4 months). In these questionnaires, the subjects were asked to evaluate their motivation, cohesion, satisfaction and self-efficacy. The presence or absence of bone overuse injuries was assessed by magnetic resonance imaging (MRI) or bone scintigraphy (BS). In addition to comparing parameters between females and males, female soldiers who sustained SF during BT (n = 18) were analyzed as a separate sub group (FSF). In general, females in a gender-integrated combat unit were more motivated and satisfied with their service than their male counterparts. Overall, the stress fractures were associated with lower ratings of motivation, self efficacy, and satisfaction expressed by the FSF sub-group. PMID- 18415120 TI - Effect of a personal ambient ventilation system on physiological strain during heat stress wearing a ballistic vest. AB - The present study was conducted in order to evaluate whether physiological strain is alleviated by a new personal cooling system (CS) consisting of a layered vest and integrated blower that generate a flow of air. Twelve male volunteers were exposed to climatic conditions of 40 degrees C, 40%RH (40/40), and 35 degrees C, 60%RH (35/60) during a 115 min exercise routine, followed by 70 min resting recovery, while wearing a battle dress uniform (BDU) and a ballistic vest, with (COOL) or without (NOCOOL) CS. The CS was able to attenuate the physiological strain levels during exercise, when compared to identical exposures without the CS. Temperature elevation, (DeltaT (re)) after 100 min of exercise, was lower by 0.26 +/- 0.20 and 0.34 +/- 0.21 degrees C in 40/40 and 35/60, respectively, (p < 0.05). Mean skin temperature (T(sk)) was lower by 0.9 +/- 0.4 and 0.6 +/- 0.5 degrees C for 40/40 and 35/60, respectively. Heart rate (HR) was not significantly different for COOL versus NOCOOL for 40/40. At 35/60, HR was lower by 10 beats per min (bpm) (p < 0.05). Physiological strain index (PSI) was 9 and 21% lower for the 40/40 and 35/60, respectively, for COOL versus NOCOOL (p < 0.05). Heat storage (S) rates were 19 and 24% lower and sweat rates were 21 and 25% lower for the 40/40 and 35/60, respectively, for COOL versus NOCOOL (p < 0.05). However, the CS was not effective in alleviating physiological strain during resting recovery with no difference in T (re) cooling rate, S, or HR drop rate between groups over resting recovery periods. The CS tested in this study was found to be an effective tool for lowering physiological strain while exercising but not during resting recovery. Therefore, the CS should be further developed in order to achieve greater attenuation of the thermal strain during exercise and improve effectiveness during rest. Overall, it has the potential to be useful for both military and sports personnel. PMID- 18415122 TI - Voltage-activated ion channels and Ca(2+)-induced Ca (2+) release shape Ca (2+) signaling in Merkel cells. AB - Ca(2+) signaling and neurotransmission modulate touch-evoked responses in Merkel cell-neurite complexes. To identify mechanisms governing these processes, we analyzed voltage-activated ion channels and Ca(2+) signaling in purified Merkel cells. Merkel cells in the intact skin were specifically labeled by antibodies against voltage-activated Ca(2+) channels (Ca(V)2.1) and voltage- and Ca(2+) activated K(+) (BK(Ca)) channels. Voltage-clamp recordings revealed small Ca(2+) currents, which produced Ca(2+) transients that were amplified sevenfold by Ca(2+)-induced Ca(2+) release. Merkel cells' voltage-activated K(+) currents were carried predominantly by BK(Ca) channels with inactivating and non-inactivating components. Thus, Merkel cells, like hair cells, have functionally diverse BK(Ca) channels. Finally, blocking K(+) channels increased response magnitude and dramatically shortened Ca(2+) transients evoked by mechanical stimulation. Together, these results demonstrate that Ca(2+) signaling in Merkel cells is governed by the interplay of plasma membrane Ca(2+) channels, store release and K(+) channels, and they identify specific signaling mechanisms that may control touch sensitivity. PMID- 18415121 TI - The role of SERCA2a/PLN complex, Ca(2+) homeostasis, and anti-apoptotic proteins in determining cell fate. AB - Intracellular calcium is a major coordinator of numerous aspects of cellular physiology, including muscle contractility and cell survival. In cardiac muscle, aberrant Ca(2+) cycling has been implicated in a range of pathological conditions including cardiomyopathies and heart failure. The sarco(endo)plasmic reticulum Ca(2+) transport adenosine triphosphatase (SERCA2a) and its regulator phospholamban (PLN) have a central role in modulating Ca(2+) homeostasis and, therefore, cardiac function. Herein, we discuss the mechanisms through which SERCA2a and PLN control cardiomyocyte function in health and disease. Emphasis is placed on our newly identified PLN-binding partner HS-1-associated protein X-1 (HAX-1), which has an anti-apoptotic function and presents with numerous similarities to Bcl-2. Recent evidence indicates that proteins of the Bcl-2 family can influence ER Ca(2+) content, a critical determinant of cellular sensitivity to apoptosis. The discovery of the PLN/HAX-1 interaction therefore unveils an important new link between Ca(2+) homeostasis and cell survival, with significant therapeutic potential. PMID- 18415123 TI - Correlations between reduced expression of the metastasis suppressor gene KAI-1 and accumulation of p53 in uterine carcinomas and sarcomas. AB - Kangai (KAI)-1 (CD82) is a metastasis suppressor gene, which belongs to the family of tetraspanin proteins. A loss of KAI-1 expression is associated with the advanced stages of many human malignancies. The present study was designed to investigate the expression pattern of KAI-1 in the normal endometrium and uterine tumors and to correlate it with the expression of tumor suppressor protein p53. KAI-1 could be found in the normal endometrium throughout the menstrual cycle. Thirteen of 42 endometrial carcinomas demonstrated moderate KAI-1 expression, but low expression of p53. Twenty-nine of 42 endometrial carcinomas showed reduced or absent KAI-1 expression, which correlated with strong expression of p53 (p < 0.001). There were significant correlations between KAI-1 expression and histological type, e.g., 93% of endometrioid carcinomas displayed a low or moderate immunostaining for KAI-1, whereas nearly all of the serous/clear cell carcinomas were KAI-1 negative (p < 0.001); tumor grading, e.g., 73% of high grade tumors showed no KAI-1 expression (p < 0.001). Most of the investigated uterine sarcomas were negative for KAI-1, whereas they displayed a strong immunostaining for p53. In conclusion, KAI-1 and p53 show inverse expression. The reduced KAI-1 expression may be the result of dysregulated p53 function and could be an important step in the endometrial carcinogenesis. PMID- 18415124 TI - Molecular detection of Leishmania parasites from whole bodies of sandflies collected in Nepal. AB - Visceral leishmaniasis is endemic in the southern part of the Terai region of Nepal. Natural infections of Phlebotomus species with Leishmania parasites in these endemic areas were analyzed by a polymerase chain reaction (PCR) amplification-based assay. A total of 401 Phlebotomus argentipes and 202 P. papatasi female sandflies were captured in the Dhanusa district from 2004 to 2006 and analyzed. It was found that 6.7% of P. argentipes, but no P. papatasi, were positive for Leishmania parasites, suggesting that P. argentipes is a major vector in these areas. The infectivity of P. argentipes with Leishmania was consistent with the infection rates reported from other areas of the world. This is the first report of naturally infected Leishmania parasites in sandflies collected from Nepal. PMID- 18415125 TI - The plot thickens: does low density affect visitation and reproductive success in a perennial herb, and are these effects altered in the presence of a co-flowering species? AB - Plants may experience reduced reproductive success at low densities, due to lower numbers of pollinator visits or reduced visit quality. Co-occurring plant species that share pollinators have the potential to facilitate pollination by either increasing numbers of pollinator visits or increasing the quality of visits, but also have the potential to reduce plant reproductive success through competition for pollination. I used a field experiment with a common distylous perennial (Piriqueta caroliniana) in the presence and absence of a co-flowering species (Coreopsis leavenworthii) in plots with one of four different distances between conspecific plants. I found strong negative effects of increasing interplant distance (related to conspecific density) on several components of P. caroliniana reproductive success: pollinator visits to plants per plot visit, visits received by individual plants, conspecific pollen grains on stigmas, outcross pollen grains on stigmas, and probability of fruit production. Although P. caroliniana and C. leavenworthii share pollinators, the co-flowering species did not affect visitation, pollen receipt or reproductive effort in P. caroliniana. Pollinators moved very infrequently between species in this experiment, so floral constancy might explain the lack of effect of the co-flowering species on P. caroliniana reproductive success at low densities. In co-occurring self-incompatible plants with floral rewards, reproductive success at low density may depend more on conspecific densities than on the presence of other species. PMID- 18415126 TI - Online monitoring of Mezcal fermentation based on redox potential measurements. AB - We describe an algorithm for the continuous monitoring of the biomass and ethanol concentrations as well as the growth rate in the Mezcal fermentation process. The algorithm performs its task having available only the online measurements of the redox potential. The procedure combines an artificial neural network (ANN) that relates the redox potential to the ethanol and biomass concentrations with a nonlinear observer-based algorithm that uses the ANN biomass estimations to infer the growth rate of this fermentation process. The results show that the redox potential is a valuable indicator of the metabolic activity of the microorganisms during Mezcal fermentation. In addition, the estimated growth rate can be considered as a direct evidence of the presence of mixed culture growth in the process. Usually, mixtures of microorganisms could be intuitively clear in this kind of processes; however, the total biomass data do not provide definite evidence by themselves. In this paper, the detailed design of the software sensor as well as its experimental application is presented at the laboratory level. PMID- 18415127 TI - Surface characteristics of grasslands in Inner Mongolia as detected by micrometeorological measurements. AB - A roving tower concept was used to compare a semi-arid grassland site in Inner Mongolia (China), which was fenced in 1979 and ungrazed thereafter (UG79) with differently grazed semi-arid steppe ecosystems. The study was conducted during three consecutive years characterised by contrasting precipitation. The different grazing intensities included continuously and moderately grazed (CG), winter grazed (WG), and heavily grazed (HG). Here, we compare the energy fluxes and surface parameters that characterise the differently managed plots. The main focus is on sensible heat flux (H), available energy (AE), surface temperature (T ( s )), and surface albedo (alpha). Systematic errors were excluded by a side-to side intercomparison of the instruments, and systematic climatic differences were minimised by the close distance between the fixed and the roving eddy covariance tower. Statistically, AE and T ( s ) were always significantly different between two simultaneously measured grazing intensities. Whereas AE was higher at UG79 in all years (mean difference of about 19Wm(-2)), T ( s ) was typically lower at UG79 (mean differences of 0.4 degrees C to about 2 degrees C). The exception was the end of the vegetation period in 2004 when T ( s ) was 0.6 degrees C higher at UG79 compared to CG. At UG79 alpha was typically significantly lower, and H was typically significantly higher. Consequently, latent heat fluxes (both as energy balance residual and directly measured) do not differ much between the different grazing intensities. It is concluded, that (1) the roving tower concept is able to detect differences due to grazing, (2) differences between the sites can be attributed to real surface differences, and (3) differences due to grazing intensities are small compared to interannual differences in surface fluxes. PMID- 18415128 TI - Highly concentrated EDTA gel improves cleaning efficiency of root canal preparation in vitro. AB - Debris and smear layer, as a product of mechanical root canal instrumentation, reduce the effectiveness of pharmacological substances to prevent post-treatment diseases and impair direct contact of filling materials with a clean dentinal surface. The aim of this in vitro study was to investigate the presence and localization of debris and smear layer via scanning electron microscope analysis after standardized root canal preparation with different chelating agents. Dentin surfaces received treatment with: (1) 15% ethylenediaminetetraacetic acid (EDTA), (2) 18.6% EDTA (3) and 24% EDTA or without any demineralizing chemicals as control. Forty vertically split human premolars were sputtered and divided into coronal, middle, and apical sections, followed by a randomized, blinded score evaluation using five scores. Pairwise comparisons of all treatment groups against a control group have been performed by Mann-Whitney U test and the Kruskal-Wallis test. Debris grades showed no significant difference between the three regions of the root canals, except for 18.6% EDTA in the central third. Smear layer and smear plug removal was concentration-dependent. Removal of the smear layer in the three areas showed that there was a statistically significant difference between all parts when using 18.6% and 24% EDTA concentrations compared with the control. The best smear layer removal in the apical region was observed using a 24% EDTA gel as chelating agent and lubricant. The usage of EDTA gel >/=18.6% presented a better cleaning regime when compared to the control group. PMID- 18415129 TI - The use of perceptual features in categorization by orangutans (Pongo abelli). AB - The extent to which categorization of natural classes in animals reflects a generalization based on perceptual similarity versus an abstract conceptual representation remains unclear. Here, two experiments were conducted to identify the perceptual features used by orangutans when categorizing pictures. In Experiment 1, subjects were trained and tested for transfer on a concrete discrimination (gorillas or orangutans vs. other primates). Analysis of performance on both positive and negative exemplars revealed that performance was best on photos with faces, particularly close-ups. Moreover, error trials did not seem to reflect instances of mistaken identity, but rather, exemplars that may have been distracting for other reasons, such as novel coloration or morphology. In Experiment 2, photos were modified to test the effects of various features. Color modifications caused the biggest decrease in performance, and eye modifications also affected performance deleteriously. Therefore, two perceptual features, namely eye regions and color, played a key role in subjects' ability to categorize. However, performance based on an underlying concept cannot be ruled out, because both of these features are highly relevant in terms of defining category membership. Although a subset of features was identified as playing a key role in categorization, these features differed depending on whether feature use was studied indirectly, as in Experiment 1, or directly, as in Experiment 2. PMID- 18415130 TI - An anatomic study for a modified technique for bypass of the external carotid artery to the proximal middle cerebral artery. AB - We aimed to evaluate whether bypass of the external carotid artery (ECA) to the middle cerebral artery (MCA) can be established by a short saphenous vein graft in order to increase the anastomosis patency. The method was performed to ten adult cadaver sides. We described a modified technique for bypass of the ECA to the M2 segment of MCA. The diameters of the vessels and graft length were measured by using an electronic micrometer. The mean diameter of the superior, middle, and inferior trunks of the MCA with trifurcation were 1.7 +/- 0.15, 2.2 +/- 0.25, and 2.0 +/- 0.2 mm, respectively, whereas the mean diameter of the superior and inferior trunks of the MCA with bifurcation were 2.1 +/- 0.2 and 2.3 +/- 0.3 mm, respectively. The mean diameter of the ECA was 3.75 +/- 0.4 mm. The mean length of the saphenous vein graft was 71.5 +/- 3.9 mm. The high-flow ECA to proximal MCA bypass using a short venous graft can supply enough blood flow to establish cerebral revascularization with a straighter route. PMID- 18415131 TI - Permeabilization of baker's yeast with N-lauroyl sarcosine. AB - N-Lauroyl sarcosine (LS), a cationic, non-toxic and biodegradable detergent readily permeabilized whole cells of baker's yeast (Saccharomyces cerevisiae). Permeabilization was carried out to increase assayable cellular catalase activity, an enzyme of great physiological and industrial importance, and to release 5'-nucleotides which find food/nutritional applications. The event of permeabilization was concentration, time and temperature dependent. Maximum permeabilization of yeast cells were observed when 1 g wet weight (0.2 g dry wt) of cells were permeabilized with 1.0 ml of 2% LS at 45 degrees C for 15 min. LS permeabilized cells showed 350-fold increase in catalase activity and the supernatant obtained after permeabilization was rich in 5'-nucleotides. LS permeabilized baker's yeast cells can be used as a source of biocatalyst and to isolate valuable by-products. PMID- 18415133 TI - Flow-network adaptation in Physarum amoebae. AB - Understanding how biological systems solve problems could aid the design of novel computational methods. Information processing in unicellular eukaryotes is of particular interest, as these organisms have survived for more than a billion years using a simple system. The large amoeboid plasmodium of Physarum is able to solve a maze and to connect multiple food locations via a smart network. This study examined how Physarum amoebae compute these solutions. The mechanism involves the adaptation of the tubular body, which appears to be similar to a network, based on cell dynamics. Our model describes how the network of tubes expands and contracts depending on the flux of protoplasmic streaming, and reproduces experimental observations of the behavior of the organism. The proposed algorithm based on Physarum is simple and powerful. PMID- 18415132 TI - The effect of electroporation pulses on functioning of the heart. AB - Electrochemotherapy is an effective antitumor treatment currently applied to cutaneous and subcutaneous tumors. Electrochemotherapy of tumors located close to the heart could lead to adverse effects, especially if electroporation pulses were delivered within the vulnerable period of the heart or if they coincided with arrhythmias of some types. We examined the influence of electroporation pulses on functioning of the heart of human patients by analyzing the electrocardiogram. We found no pathological morphological changes in the electrocardiogram; however, we demonstrated a transient RR interval decrease after application of electroporation pulses. Although no adverse effects due to electroporation have been reported so far, the probability for complications could increase in treatment of internal tumors, in tumor ablation by irreversible electroporation, and when using pulses of longer durations. We evaluated the performance of our algorithm for synchronization of electroporation pulse delivery with electrocardiogram. The application of this algorithm in clinical electroporation would increase the level of safety for the patient and suitability of electroporation for use in anatomical locations presently not accessible to existing electroporation devices and electrodes. PMID- 18415134 TI - Neuroevolution and complexifying genetic architectures for memory and control tasks. AB - The way genes are interpreted biases an artificial evolutionary system towards some phenotypes. When evolving artificial neural networks, methods using direct encoding have genes representing neurons and synapses, while methods employing artificial ontogeny interpret genomes as recipes for the construction of phenotypes. Here, a neuroevolution system (neuroevolution with ontogeny or NEON) is presented that can emulate a well-known neuroevolution method using direct encoding (neuroevolution of augmenting topologies or NEAT), and therefore, can solve the same kinds of tasks. Performance on challenging control and memory benchmark tasks is reported. However, the encoding used by NEON is indirect, and it is shown how characteristics of artificial ontogeny can be introduced incrementally in different phases of evolutionary search. PMID- 18415136 TI - [Not Available]. PMID- 18415138 TI - [Not Available]. PMID- 18415137 TI - [Not Available]. PMID- 18415140 TI - [Not Available]. PMID- 18415139 TI - [Not Available]. PMID- 18415142 TI - [Not Available]. PMID- 18415141 TI - [Not Available]. PMID- 18415143 TI - [Not Available]. PMID- 18415144 TI - [Not Available]. PMID- 18415145 TI - [Not Available]. PMID- 18415146 TI - [Not Available]. PMID- 18415148 TI - [Editorials.]. PMID- 18415147 TI - [Not Available]. PMID- 18415149 TI - [Not Available]. PMID- 18415150 TI - [Limitations of and new options with neurophysiological methods in migraine research and diagnosis : Electroencephalographicmethods.]. AB - In parallel with the development of the various electrophysiological methods now available, the clinical syndrome of migraine has been repeatedly analyzed with the aid of these tools, in attempts to track its course with reference to electrophysiological parameters. A survey of the international literature reveals the following EEG findings. (1) In patients with different forms of headache without structural lesions the distribution of normal and abnormal EEG recordings during headachefree intervals resembles that seen among healthy subjects. (2) In common migraine, nonfocal and nonspecific abnormalities are reflected in up to 65% of all EEG recordings during the migraine-free interval. (3) Focal EEG abnormalities during a migraine attack are relatively rare, generally occurring only during migraines with transient focal neurological disturbances. (4) It is very rare for there to be permanent focal changes that are apparent during both pain-free intervals and migraine attacks. In the presence of such changes very careful diagnosis is necessary to exclude symptomatic lesions. (5) In up to 53% of migraineurs a few minutes' hyperventilation leads to generalized slow, wide amplitude, sometimes episodic waves in the EEG. Similar changes also occur in healthy subjects, but are less common, less intense and of shorter duration. (6) There is a statistically significant correlation between the occurrence of a migraine and EEG activation by photic stimulation at frequencies over 20 Hz. (7) No unequivocal results obtained in large series of patients with train mapping are yet available. PMID- 18415151 TI - [Documantation of pain-related data in outpatients with a specially designed pocket computer (Rating Box).]. AB - The documentation of illness-related data, e.g. repetitive recordings of pain parameters, medication or mood, is commonly accomplished by the use of questionnaires. Several disadvantages for both the patient and experimentor related to this method can be avoided by the application of specially designed data-loggers. The use of commercially available portable pocket computers is usually complicated because of the miniature full-range keybords. This paper describes a completely new design, which relies on the use of only four keys. All operations are imaged on an LCD display (4 lines with 16 signs). The Rating Box allows the presentation of visual analogue scales (VAS) of different step widths, movement of cursors, forced choice and multiple choice decisions, as well as the input of decimal numbers. Up to 11 scale types can be deliberately combined in a series of up to 27 items. The combination can be individually designed and edited by the experimentor. The results are stored together with the input time and date. Actual inputs are not influenced by comparison to earlier input, since data recall can only be accessed by the experimentor. The use of the Rating Box by a patient is called up by signal tones at individually given times of day. Additional inputs are possible when required. Warnings are presented for general malfunctions. The results can be transmitted to a personal computer and processed statistically or to daily or weekly profiles of mood, pain or disability scores, for example. A pilot study in elderly patients (42-76 years) with pain due to cancer gave promising results. The Rating Box was well accepted as opposed to a questionnaire. Nine out of 12 patients preferred the Rating Box both methods were judged to be equivalent by two patients and only one regarded the use as difficult and thus preferred the questionnaire. In addition, 5 out of 12 patients confessed by inquiry to have filled in the questionnaire forms retrospectively. This possibility is in principle excluded by the Rating Box. PMID- 18415152 TI - [The International Headache Society's new headache classification system.]. AB - A completely new classification system for headache disorders, cranial neuralgias and facial pain has been prepared by a headache classification committee working within the International Headache Society (IHS). The committee is made up of an international body of experts, who spent almost 3 years on the preparation of the classification, publishing it in 1988 (Cephalalgia 8 Suppl 7ratio1ff., 1988). The German translation appeared a year later (Nervenheilkunde 8ratio161-203, 1989). The new classification system is structured in 13 main groups, each with finer divisions into subgroups. Preliminary experience with application of the new classification in 243 patients is reported. The advantages and the problems encountered are discussed. On balance, the new IHS classification is proving to be a very useful basis for the diagnosis of painful conditions affecting the head and face. It is indispensable for a modern headache clinic. PMID- 18415153 TI - [Pain therapy with implanted medication pumps for chronic pain.]. AB - Epidural or intrathecal opiate analgesia, combined with bupivacain by means of an implanted pump, represents a possibility for providing good pain management for cancer patients as well as other chronic pain patients. Several indications, for implantation of a percutanously refillable pump are demonstrated in 27 patients. Twenty-four patients were treated with epidural and 3 with intrathecal catheters. Nineteen patients were suffering from chronic pain, and 8 had pain because of cancer. Four patients with chronic pain have been treated with continuous epidural opiate analgesia by means of an implanted pump for more than 2 years and 1 patient for more than 5 years. In the course of 2 years there has been no significant increase in the daily dose of buprenorphin given epidurally to patients with chronic pain. There were no addiction problems with opiates given epidurally or intrathecally by means of implanted pumps. Because of a 13% complication rate, pumps and epidural or intrathecal catheters should only be implanted by an experienced team. PMID- 18415154 TI - [Continuous administration of opiates with implantable drug delivery systems in patients with intractable non-malignant pain.]. AB - The use of implantable systems for intrathecal administration of opioids in chronic pain of non-malignant origin is a controversial subject. Opioid therapy is reserved mainly for pain patients with malignant disease and reduced life expectancy. The main reasons for this restricted range of indications of chronic subarachnoid administration of opioids are fear of addiction and the build-up of tolerance. During July 1988 and April 1990 we treated 60 patients suffering from pain of non-malignant origin with continuous opioid infusion by implanted pumps. Wishing to find whether opiates can relieve deafferentation pain, we subdivided the different pain syndromes into three groups according to their pathophysiology: nociceptive, neurogenic/neuropathic, and deafferentation pain. After a follow-up period of 11.5+/-7.1 months 47 patients were evaluated. Pain intensity according to the visual analogue scale was reduced in a mean of 79.4% of the patients. Activity level and mood scores as pain-associated parameters were both significantly increased after therapy. Analysis of the McGill Pain Questionnaire reveals that the improved quality of life is attributable to a reduction of affective pain perception more than to sensory discrimination. The best results in terms of pain reduction (82.5%) were obtained in the group of patients with deafferentation pain. This is in contrast to reports in the literature. It seems that neuropathic and deafferentation pain syndroms are susceptible to intrathecal opioids. The initial daily average dose of morphine was 2.6 mg/day, increasing to 6.1 mg/day after 25 months without the development of major tachyphylaxis. the administration of intrathecal opioids by means of implantable systems is justified in carefully selected patients with chronic non malignant pain. This method should be applied in preference to destructive neurosurgical treatments. PMID- 18415155 TI - [Not Available]. PMID- 18415157 TI - [Not Available]. PMID- 18415156 TI - [Not Available]. PMID- 18415158 TI - [Not Available]. PMID- 18415159 TI - [Not Available]. PMID- 18415161 TI - [Not Available]. PMID- 18415160 TI - [Pathophysiological mechanism of reflex sympathetic dystrophy.]. PMID- 18415162 TI - [Not Available]. PMID- 18415163 TI - [Neurophysiology of nociception and pain in deep tissue (skeletal muscle, tendon, joint, connective tissue).]. AB - Present-day hypotheses about the origin of pain in deep tissues are based on the idea that pain is anindependent sensation with its own specialized apparatus of sensors, conduction pathways and centers. The sensors are callednociceptors ornocisensors, and the neuronal structures they activate are thenociceptive system. Accordingly, the reception, conduction and central nervous processing of noxious signals together are termednociception. All parts of the nociceptive system may be affected by pathophysiological processes which in turn may modify appreciably the perception of pain. Variouscomponents of pain are activated by noxious signals. The contributions of the sensory, affective, autonomic and motor components to the evaluation (cognitive component) and expression (psychomotor component) of the pain vary, depending on the nature of the pain. Again, all components may be modified by pathophysiological processes. It is also pointed out that an important aspect in evaluating pain is its duration, and various types ofacute and chronic (persistent, recurrent) pains and their mode of origin are discussed. Theneurobiology of joint pain is used to exemplify the processes of excitation (transduction) in nociceptors (nocisensors), the subsequent step of transformation, the peripheral conduction and central processing of noxious signals. Special emphasis will be given to the new finding that healthy tissues contain nociceptors with threshold so high that they cannot be excited by acute noxious stimuli ("sleeping" nociceptors). However,sensitization of these nociceptors as a consequence of pathological tissue alterations (e.g. by inflammation) will "awaken" them. Sensitization is probably brought about by algesic substances (mediators of inflammation, e.g. prostaglandins, bradykinin). The similarities and dissimilarities of joint pain versus other deep pain (originating from muscles, tendons or bones) are pointed out. The increase in sensory inflow resulting from the sensitization of peripheral nociceptors leads subsequently to acentral component in the sensitization which in turn modifies the perception of pain under these circumstances. Various processes such as expression of C-fos-like-protein, modification of the spinal composition of endogenous opioids, and release of neuropeptides may be involved in this newly detected phenomenon. Its possiblerelation to the development of chronic pain states is discussed. Finally an account is presented of pain produced by excitation of the nociceptive system proximal to the nociceptors. These pain states include pain resulting from pathophysiological impulse generation in nociceptive fibers (neuralgia orneuralgic pain) which usually projects into the region containing the sensory endings of these fibers (projected pain). Furthermore, brief descriptions of pain due to spinal root compression and ofcentral pain arising from various sites of the central nervous system are given. PMID- 18415164 TI - Symptomatology and clinical pathophysiology of myofascial pain. AB - Myofascial pain syndromes, fibromyalgia, and articular dysfunctions may all be contributing to our patients' ubiquitous musculoskeletal pain problems that generally are poorly understood and poorly managed. Thepectoralis minor myofascial pain syndrome, for example, results from trigger points (TrPs) activated by stress overload of the muscle. Symptoms of pain referred to the shoulder and ulnar aspect of the arm and forearm, and of pain on reaching around and behind the body, are characteristic. Findings include restricted stretch range of motion and some weakness of the muscle, taut bands of muscle fibers, and focal trigger point tenderness of each taut band on palpation. Snapping palpation at the TrP elicits a local twitch response (LTR). The increased muscle tension of a pectoralis minor syndrome commonly entraps the lower trunk of the brachial plexus, producing symptoms of a cervical radiculopathy. PMID- 18415165 TI - [Pain of the joint of inflammatory diseases pathobiochemistry and pharmacology.]. AB - Pain is the leading symptom of inflammatory joint diseases. It is immediately caused by the release of prostaglandins (and potentially leukotrienes) from cells of the inflamed tissues, which sensitizes the pain receptors. The synthesis of these mediators depends on the activation of infiltrated inflammatory cells, as well as recruitment of tissue born cells, predominantly by the inflammatory cytokines Interleukin-1 (IL-1) or tumor necrosis factor (TNF). In chronic diseases such as rheumatoid arthritis the inflammatory reaction is initialized and perpetuated by (auto)-immuno reactions. The associated chronic pain is thus the end point of a complex multi-level disease process. The hierarchy of these regulatory processes is mirrored by the pharmacological interventions. Inhibitors of the key enzyme of prostaglandin synthesis, cyclooxigenase, such as the non steroidal anti-inflammatory drugs (NSAID) are immediately analgetic. Anti inflammatory drugs as the glucocorticoids predominantly decrease the synthesis of cytokines, and thereby the stimuli leading to prostaglandin synthesis. Together with a decrease of the synthesis of arachidonate metabolizing enzymes this leads to correction of pain. Although not directly analgetic, immunosuppressive drugs, too, by decreasing the immune reaction dependent inflammation, contribute to pain relief. PMID- 18415166 TI - [Clinical diagnosis and documentation of chronic-rheumatic pain syndromes the state of the art 1990.]. PMID- 18415168 TI - [Local and regional therapy of pain syndromes related to the musculosceletal system.]. PMID- 18415167 TI - [Not Available]. AB - Oral medication is the simplest way in treatment of chronic pain. For cancer pain oral analgesics are efficacious in more than 90% of the patients. When a causal therapy of pain (e.g. chemotherapy, operation) fails an analgesic ladder with oral analgesics is instituted. This ladder starts with a non-narcotic analgesic in a sufficient dose. The regular dose of acetylsalicylic acid or paracetamol is 4 g daily. When this dose does not work sufficiently, a weak opioid (e.g. dihydrocodeine) is given concomitantly at an individual dose. When the weak opioid fails, strong opioids are given (e.g. morphine). The drugs should be given by mouth whenever possible. The most important point is the regular application according to a time-schedule. This time-schedule is related to the action time of the drug. Patients with severe vomiting or dysphagia can receive a continuous subcutaneous infusion. These measures are based on recommendations of the WHO.The same medications can be employed in patients with chronic non-malignant pain, provided that all other conventional measures in pain treatment fail. However, many states of pain are not opioid-responsive. Pain related to the sympathetic nervous system is more responsive to antidepressants than to opioids or NSAID. Neuropathic pain as in trigeminal neuralgia responds to anticonvulsants. Pain from muscle spasm is better controlled by muscle relaxants than by analgesics. Bone pain is more sensitive to NSAID than to any other drug.In any state of pain the response to the different groups of drugs should be evaluated first. Then a stepwise pharmacological approach should be performed. In most cases pain can be treated effectively by oral drugs. PMID- 18415169 TI - [Advances in the physical therapy of pain in rheumatology.]. AB - The analgesic effectiveness of physical therapy in rheumatology is dependent on the differentiated clinical picture (joint, soft parts, spinal column, musculature) and on the differentiated therapeutic remedy (intensity, duration of single treatments, frequency, duration of therapeutic series). Physico- and kinesitherapy can be distinguished with regard to objective and subjective doses; manual therapy is between them. During the last decade, particular advances were achieved in the field of medical gymnastics (muscular stretching technique), patients' schools (back school) and of cryotherapy (cold-air therapy). The pulsed high-frequency therapy and particularly laser therapy, magnetic-field therapy and high-voltage therapy are critically judged. The problem "rheumatism and pain" mainly exists at the level of "chronic"; diagnostics of movement function (articular and muscular functions) on one hand and dosage of therapeutic remedies (drugs and remedies of physiotherapy) on the other are the guidelines. Course (rehabilitation) as well as onset (prevention) of rheumatic clinical pictures determine the further strategy of pain therapy. PMID- 18415170 TI - [Psychological pain treatment in rheumatic patients.]. AB - Psychodynamic concepts postulate a psychogenesis of physical pain proposing several assumptions about the conversion of mental suffering into physical pain. Behavioural concepts, on the other hand, emphasize psychological conditions as risk factors for chronicity and describe psychological reactions to chronic pain. Patients with painful diseases and inadequate coping strategies very often display symptoms of anger, anxiety, or depression. Recently, the use of group therapy aimed at enhancing patients' ability to cope with disease-related stressful events has become widely accepted in behavioural medicine with a focus on pain-management procedures. Strategies for the improvement of coping with pain are based on behavioural, psychophysiological, and cognitive principles. The behavioural view conceptualizes pain as a behavioural problem with regard to facial and bodily expressions of pain, decreased physical and mental activity, and the consumption of pain medication. Operant conditioning is used to discourage pain behaviour and reinforce well-behaviour. The physiological concept stresses the vicious cycle of pain, increased muscle tension, and emotional reactions. Relaxation procedures are introduced in order to reduce excessive muscular activity in targeted muscles. The cognitive approach emphasizes the effect of information-processing on pain experience. Cognitive distortions are identified, and self-control management is encouraged. Having taken all of these aspects into consideration, we developed a cognitive-behavioural treatment programme in a group setting format with components of relaxation, cognitive restructuring, and the promotion of well-being. Subjects included in the study were given diagnoses of low back pain, tension headache, rheumatoid arthritis, and ankylosing spondylitis. Treatment effects in different diagnostic groups were compared to each other, supporting the assumption that pain reduction is greatest in low back pain and least in ankylosing spondylitis. Subjects with inflammatory rheumatic diseases showed some improvement in self-reported physical complaints and in their feelings of well-being. PMID- 18415171 TI - [Not Available]. PMID- 18415172 TI - [The artist as a pain patient. An investigation on pain in the musculoskeletal system in musicians and their expectations regarding pain therapy.]. AB - 74 pop/rock/jazz musicians and 100 classical musicians were investigated by means of a specially designed questionnaire. Several social, musical, pain-, and health related questions were asked. Pain of the musculoskeletal system was the main complaint of musicians in both group (75% classical; 64% rock-pop-jazz). Classical musicians suffer most from neck pain (35%) and the other group mostly from shoulder pain (30%). Specific instruments showed specific muscolosceletal pain patterns. 55% of the musicians in the classical field were treated by an orthopedic surgeon, whereas 43% of rock-pop-jazz musicians chose no therapy, although they suffered from pain. Only 32% of the classical musicians showed good compliance; 64% believe that medical therapy is not adjusted to the needs of musicians. 98% of the classical musicians and 89% of rock-pop-jazz musicians wish to have a doctor who is specially trained to deal with the needs of musicians. PMID- 18415173 TI - [Pain complaints among inhabitants of Lubeck: results of a population-based epidemiologic study.]. AB - The paper reports the results of a population-based pain survey in Lubeck, a city of 210,000 inhabitants in the northern part of Germany. Data were collected from 308 respondents (systematic sample aged 26-75 years) by mailed questionnaires (response rate: 80%). Subjects indicated on a list of 11 pain conditions whether they suffered from these kinds of painever, during the past 6 months, or "today". The prevalence of the 11 pain conditions ranged from 19% to 70% (lifetime prevalence), 14% to 56% (6-month prevalence), and 4% to 29% (point prevalence). Rheumatic pain (e.g., back pain) prevailed; higher prevalence figures were found for females for several pain conditions. Respondents who suffered from pain "today" (n=157) or during the preceding 6 months (n=255) often reported two or more kinds of pain. Ten percent of the respondents reported that they had never experienced any of the 11 pain conditions, 17% did so with respect to the past 6 months, and 49% were free of pain on the day they completed the questionnaire. The classification of pain sufferers according to the number of disability days in the past 6 months showed that three-quarters of them reported 0-6 disability days. However, 8% of the total sample reported more than 14 pain-related disability days and were classified as being affected by pain to a sociomedically relevant degree. Approximately half of the subjects who reported pain in the past 6 months did not consult a physician. This proportion decreased considerably in subgroups with more than 6 disability days. PMID- 18415174 TI - [Multi-dimensional evaluation of pain perception and its importance for the behavior of patients with chronic low back pain.]. AB - A consecutive series of 98 patients presenting at an orthopedic outpatient clinic with chronic low back pain of at least 6 months' duration and with no organic findings (ruled out by clinical and radiological examination) were evaluated by means of a questionnaire which included the constructs "patient history," "pain related restrictions," and "depression." Pain perception was evaluated with an adjective list revealing four main factors: two affective factors, i.e., "suffering from pain" and "anxiety," and two sensory factors, i.e., "acuteness" and "rhythmics of pain." The two affective factors (as against the sensory factors) subsequently influence the degree of pain intensity (measured with a visual analog scale), the patient's history and the patient's perceived impairment of daily life. Depression (von Zerssen scale) correlated with pain factors only when the whole range of pain factors was considered. The implications for treatment in patients with a high score for affective factors in the adjective list (indicator for a low success rate with traditional therapy) are discussed. PMID- 18415175 TI - [Physiodiagnostic indicators in tension headache.]. AB - Practitioners often rely on physiodiagnostic indicators to corroborate the hypothesis of a muscular origin of headache. Although these indicators have been widely applied, their reliability and validity have seldom been tested empirically in headache sufferers. In a controlled double blind study, two trained raters palpated muscle tension and latent and active myogeloses of the left and right trapezius and sternocleidomastoideus muscles and measured passive head rotation flexibility. There were 28 subjects in the study, 14 suffering from tension headache and 14 matched controls. In spite of the training, interrater reliability of ratings concerning degree of muscle tension and number of latent myogeloses proved to be too low to justify subsequent use of these indicators. In keeping with the hypothesis, the various parameters of active myogeloses very clearly differentiated between the experimental groups. The hypothesis turned out not to be true for the parameters of head rotation flexibility. In subjects suffering from tension headache, no correlations could be found between the number of myogeloses of the right trapezius muscle and parameters recorded in long-term EMGs of this muscle, and no correlations could be found between the total number of myogeloses and the chronicity of headache. PMID- 18415176 TI - [A follow-up study of chronic headache patients in a neurological outpatient department.]. AB - The chronic headache patients in our neurological outpatient department treated between 1985 and 1987 were retrospectively studied. One-third (n=44) were examined and questioned about the efficacy of treatment. Initial treatment in the outpatient department had been at least 2 years before the study, thus allowing evaluation of the long-term course of the illness. Age distribution, sex ratio, distance to the therapy center, duration of illness, and the extent and results of the diagnosis are reported. The most common form apart from migraine and tension headache was drug-induced headache. The range of medical and non-medical therapy applied is presented. The therapeutic response (free of pain/distinct improvement) was 43.2% directly following therapy, 40.9% after 1 year, and 34.1% 2 years after therapy. Although the results correspond with those of other studies, they also indicate, however, a high level of resistance to therapy. Long term medical side effects, as well as psychosocial factors, especially appear to exacerbate chronic headache in the non-responders. Further investigations are necessary to explore this factor. The practical consequences are discussed for outpatient therapy of headache patients. In addition to the exact clinical classification of the headache syndrome, we consider it important to identify the exacerbating factors before starting multidimensional therapy. Adequate consideration of the non-medical therapeutic elements should be ensured in such structures headache therapy. A uniform classification of headache and records of the course in the form of headache diaries are essential for comparing the results. A sufficiantly long post-therapy observation period should be allowed in order to facilitate evaluation of the therapeutic response. PMID- 18415178 TI - [Spinal neurostimulation for the treatment of chronic pain: changes in indications and patient selection after 19 years' experience.]. AB - After the initial clinical reports of Shealy 1967 dorsal column stimulation (DCS) was first introduced in Germany by Krainick (Freiburg) and Winkelmuller (Hannover) in 1972. At first, the success rate in unselected patients was unsatisfactory. The results improved with careful patient selection and better technical equipment allowing preliminary testing procedures before definitive implantation. The authors' own results in 335 patients treated by intermittent spinal cord stimulation (SCS) with implanted devices between 1972 and 1989 show that long-term beneficial effects can be obtained in pain of neurogenic origin rather than in nociceptor pain. Guidelines for the use of SCS were proposed by the German Society of Neurosurgery in 1990. The best indications and target group are cases with radicular low-back pain after failed back surgery, stump and phantom pain, pain states following partial lesions of brachial/lumbar plexus and peripheral nerves, sympathetic dystrophy and rest pain in peripheral vascular disease (PVD). Possible indications for SCS are pain after incomplete lesions of spinal cord or cauda equina, postherpetic neuralgia, sclerodermia and PVD. Failures must be expected in pain states related to progressive malignant disease and complete deafferentation after spinal lesions or root avulsion. PMID- 18415177 TI - [Sustained-release dextropropoxyphene.]. AB - Dextropropoxyphene is a mild opioid analgesic whose analgesic potency corresponds to that of acetylsalicylic acid and paracetamol. It has a similar analgesic effect to codeine but also a considerably lower addiction and dependence potential. Dextropropoxyphene is a therapeutic alternative to other weak opioids such as codeine or dihydrocodeine. In the case of absolute intolerance of non steroidal anti-inflammatory agents, their analgesic effect can be replaced by that of dextropropoxyphene. In case of relative intolerance, i.e. occurrence of non-tolerable side-effects, the dose of non-steroidal anti-inflammatory agents can be kept low by additional administration of dextropropoxyphene, which simultaneously enhances analgesia. Analgesics are prescribed according to a definite time schedule for the long-term treatment of chronic pain. The oral route of administration is preferred since it enables the patient to be independent of the nursing staff. Sustained-release drugs with a duration of action of at least 8 h are used in preference to other preparations. Sustained release dextropropoxyphene provides analgesia for 8-12 h. Sustained-release dextropropoxyphene clearly differs from non-sustained-release dextropropoxyphene in its pharmacokinetics. Repeated administration of the sustained-release form at the therapeutically recommended intervals does not lead to cumulation, and the risk of accidental overdosage is extremely low. Intoxication can only occur after simultaneous ingestion of alcohol or other centrally depressant substances or in the presence of hepatic and/or renal failure. Sustained-release dextropropoxyphene is a sensible and undeniable alternative for the second stage in the analgesic ladder of chronic pain therapy. PMID- 18415179 TI - [Technical development of spinal neurostimulation.]. AB - The rationale for the use of spinal cord stimulation (SCS) in the treatment of chronic pain arose from Melzack and Wall's gate theory of the control of pain (1965). Originally electrodes were placed directly on the spinal cord via open operation, while now they are placed by means of direct puncture the epidural space. In most cases, one of two different systems is used: (1) semi-implants, consisting of an implantable electrode, connection cable and receiver, and externally worn stimulator with antenna; (2) full implants, consisting of an implantable electrode, connection cable and a stimulator, which is also implanted. The implantation of an SCS system is a surgical procedure, which requires the highest standards in asepsis. The operation and its complications and ways of avoiding them are described. PMID- 18415180 TI - [Spinal baclofen in trigeminal neuralgia. A case report.]. AB - Oral baclofen, the most often prescribed antispastic drug, has been shown to be effective in trigeminal neuralgia. Spinal application of baclofen leads to a complete supression of spasticity even in cases in which no previous oral antispastic medication achieved a response. Since these results suggest that spinal administration of baclofen is superior to oral baclofen, spinal injections of this drug (100 mug) were tested in a patient with severe trigeminal neuralgia that was unresponsive to oral medication. This was followed by complete suppression of the neuralgic symptoms, which however reoccurred if baclofen administration was stopped. Spinal baclofen has also been effective during long term infusion with an implanted pump for more than 1 year. Spinal baclofen infusion seems to be a possibility in the treatment of severe trigeminal neuralgia resistant to other forms of therapy. PMID- 18415181 TI - [Pain therapy with opioids: experience and thoughts of a general practitioner.]. AB - Medical treatment with effective opioids for patients who suffer chronic pain is greatly lacking in Germany, as is supported by the documentation from Sorge and Zenz. The author comments on this documentation and adds an account of his own experience with the provision of opioid prescriptions over a period of almost 6 years. He asserts that the number of patients who suffer from pain has increased and argues for extending the indications for opioid therapy to include noncancer patients, giving reasons why pure morphine preparations are to be preferred. Finally, the author expresses his belief that only an expanded and appropriate application of today's concepts regarding treatment with analgesics and opioids will be able to clear the way for a liberalization of the laws regulating the prescription of opioids. PMID- 18415182 TI - [Not Available]. PMID- 18415183 TI - [Not Available]. PMID- 18415184 TI - [Not Available]. PMID- 18415185 TI - [Art therapy in chronic pain.]. AB - In the therapy of chronic disease and functional disorders, art therapy is considered to be of increasing importance. The idea behind this type of therapy is that the stimulation of creative activities promotes the healing process and rehabilitation. Music therapy in particular has a long tradition in the treatment of pain and health disorders. In patients, whose perception of disease and treatment expectation is determinated by the idea that their pain results from physical causes alone, compliance in music therapy is usually better than direct psychotherapeutic approaches. In addition to music therapy, the creative arts, creative dance and movement-based creative therapy have also been used in the treatment of patients with chronic pain. A model of chronic pain as a changeable phenomenon of consciousness which depends on subjective perception, communication and coping is the basis for understanding art therapy. The conceptual framework of art therapy offers various explanations for the integration of these forms of therapy in complementary, supportive pain management programs: (1) enhancing the activity level and creative capacity as a healing source; (2) stimulation of positive emotional experience; (3) experiencing social communication and interaction; (4) facilitating projective coping; (5) stimulation of imaginative experience and awareness; (6) promotion of suggestive elements. Anecdotical experience indicates that there could be a broad field for the use of art therapy in pain management programs. The need to validate this form of therapeutic approach by appropriate methodological studies and well-documentated single case series is emphasized. PMID- 18415186 TI - [Pain in the fetus: neurobiological, psychophysiological and behavioral aspects.]. AB - Until a short time ago, the view prevailed worldwide that children were less sensitive to pain than adults, and such operations as circumcision were performed in babies without adequate anesthesia or analgesia. This view is now considered a misconception, as psychophysiological and behavioral studies show that even neonates have a well-functioning nociceptive system. Nociception generally refers to the neural and sensory aspects of pain, which do not necessarily include conscious experience. There is no discontinuity in the development of the nervous system during birth, and therefore it can be concluded that the fetus is also responsive to noxious stimuli. The question arises as to the stage of ontogeny of the human at which nociceptive behavior begins. Literature on the fetal nervous system reveals that the first signs of somatosensory system function occur at week 7 of gestation and at week 22 the synaptic connection from the nervous periphery to the somatosensory cortex is becoming established. During this period, motor behavior matures, from stereotyped reflexes to spontaneously generated complex motor patterns reminiscent of the repertory of voluntary movement. From week 22 onward the electroencephalogram (EEG) shows increasingly more varied patterns, and sleep-wake states can be discerned after week 30 of gestation. Somatosensory evoked cortial potentials have been recorded from gestational week 28 onward. Substance P, a neuropeptide associated with pain in the adult nervous system, is present in the fetal spinal cord as early as week 12 of gestation, while the antinociceptive opioid peptide enkephalin does not appear until week 24. From week 15 onward, opioid peptides such as beta-endorphin appear in the pituitary; their release becomes sensitive to environmental stimuli from about week 20 onward, which can be considered the onset of pituitary stress responses. In particular, parturition and abortion induced the release of opioid peptides. Studies of conditioned behavior show that the fetus has the ability to learn. It has been hypothesized that the fetus and neonate possess a procedural memory, which is not transferred to the language-based memory of later phases of life. Learning and memory are the most essential elements for the construct of "consciousness." Therefore, a primitive type or level of consciousness might exist in the fetus. Thus, a considerable range of sensorimotor function, including memory, develops during fetal life. Anatomical, physiological and behavioral data suggest that the nociceptive system is included in this development. Although we cannot be sure at present whether the fetus consciously experiences pain, beyond the protective nociceptive behavioral responses, anesthesia should be used for invasive procedures to protect the fetus and its nervous systems. PMID- 18415187 TI - [Pain therapy in cancer pain patients: control of therapy with the Potsdam pain questionnaire.]. AB - 154 cancer pain patients were treated from 1988 to 1990 according to the three step analgesic ladder (TSAL) recommended by the WHO. The patients were assigned into 4 groups: WHO-I: patients were treated only with non-opioids, WHO-II: patients received a combination of non-opioid plus a weak opioid, WHO-III: patients were treated with strong opioids or a combination of a strong opioid plus a non-opioid; and the 4th group consisted of patients receiving a strong opioid epidurally. Our results, using the TSLA and comparing then to the pretreatment status, show complete or acceptable cancer pain relief among the patients as follows: in WHO-I: 44%, in WHO-II: 55%, in WHO-III: 63%, and in the 4th group 70%. The use of the TSAL resulted not only in a reduction of the pain level, but also in an increase of the quality of the life for these cancer pain patients. Also the mean duration of the patients' sleep was increased by more than one hour. An improvement of mood was observed, and the stabilisation of mood lasted longer. PMID- 18415189 TI - [Predicting the outcome of diskectomy.]. AB - The outcome of disk surgery in 40 consecutive patients was predicted by pre treatment assessments of sociodemographic and psychological variables and findings in a standardised orthopaedic and neurological examination. The pre surgery variables that proved to be associated with outcome criteria six months post surgery by means of a multiple stepwise regression procedure were selected for discriminant analyses, using three outcome criteria: functional status, patient evaluation of the outcome, and vocational rehabilitation. The correct classification rate was 85%, 77.5%, and 90%, respectively. No prediction was possible for postoperative pain behaviour and postoperative orthopaedic and neurological status. Significant predictors were time off work before surgery, active search for information about disease and surgery, presence of conditions that reinforce pain behaviour, and cognitive variables indicating helplessness. PMID- 18415188 TI - [Physical exercise, endogenous opiates and pain regulation.]. AB - A series of studies with humans as well as experiments carried out on animals have shown that physical exercise leads to temporary hypoalgesia. Reduced sensitivity to pain is not only demonstrable after long-distance exercise (such as a marathon run) but also during and after intensive physical exercise on a laboratory ergometer. In a double blind study (20 mg naloxone versus placebo) experimental pain thresholds (electrical intracutaneous finger and dental pulp stimulation) and plasma hormone levels (beta-endorphin, cortisol, and catecholamines) were measured in ten healthy athletic men before, during, and after physical exercise on a cycle ergometer. A significant pain threshold elevation during exercise was found for finger (Anova,p<0.004) and dental pulp stimulation (p<0.01). Hypoalgesia remained present after exercise was stopped and the initial pain threshold level was returned to approximately 60 minutes after the exercise. The subjective magnitude estimation of suprathreshold stimuli was significantly reduced (p<0.001) after exercise. Naloxone failed to affect pain thresholds and plasma beta-endorphin did not correlate significantly with pain thresholds. The cause of the exercise-induced hypoalgesia is probably an activation of central pain inhibitory mechanisms by the "stimulus" of physical exercise (stimulation-induced analgesia). Central pain inhibitory systems are probably thereby activated by the stimulation of afferent nerves endings (group III and IV) in the skeletal muscle. The same trigger mechanism also plays a role as a release stimulus for hormones which are secreted in increased measure during physical exercise (catecholamines, pituitary hormones). Plasma beta-endorphin is probably not directly involved in the exercise-induced hypoalgesia but is rather a "marker" for the activating of central analgesia mechanisms. PMID- 18415191 TI - [Not Available]. PMID- 18415190 TI - [Not Available]. PMID- 18415192 TI - [The right to pain relief.]. AB - In the Federal Republic of Germany the medical and therapeutic methods available for combatting the most severely painful states are not always fully exploited. The medical legislation currently being developed gradually gives the treatment of pain the importance it deserves according to patients' needs. The legal position to be borne in mind in pain therapy is presented. PMID- 18415193 TI - [Outpatient care of patients with chronic painful states. A contribution to the quality of life.]. AB - The care of severely ill patients, whether in hospital, in residential homes or in their own homes, should be characterized by humanity and dictated by efforts to make life worth living for the person concerned. Preservation of the quality of life should be paramount if the doctor can no longer effect a cure. Being cared for at home in the domestic setting is an enhancement of the quality of life in itself. A change in surroundings and in the daily round is particularly burdensome for a person who is ill. Close contact with members of the family and with friends and neighbours is particularly important in this phase of life. A sick person's quality of life is what the legislator had in mind when domiciliary care was given priority over residential care at the time of the legal changes to reform the health care system. PMID- 18415194 TI - [Opioids in "non-malignant" pain-results of long-term treatment in patients with rheumatic disease.]. AB - The oral administration of strong opioids like morphine is a very effective treatment in cancer pain. However, these analgesics are rarely prescribed for patients suffering from severe "non-malignant" pain. We examined the effects of oral opioids (morphine sulphate tablets, buprenorphine and levomethadone) given to patients with intractable rheumatic pain, which were refractory to other therapeutic measures. The origin of pain was inflammation or a degenerative lesion of the spine. Within a period of more than 3 years, 12 patients were treated accordingly. In 9 patients we could achieve sufficient pain relief, two of them showing improvement only after having changed the initially prescribed drug. We had to stop opioid medication in two patients because of side-effects and, moreover, in one patient because of failure to produce analgesia. 775 weeks of treatment were documented until December 31th, 1990, with an individual duration ranging from 11 to 145 weeks. It was necessary to increase the dose of morphine in the course of treatment of one patient, who is up to now being treated for more than 77 weeks. In all other patients the doses were either stable or varied. No severe side-effects such as respiratory depression were associated with long-term opioid therapy. Constipation was observed in 4 patients, nausea in two patients and urinary retention in one patient. These side effects could be well treated by an additional medication. No drug abuse, dependence or tolerance were observed. Strong opioids are not analgesics of first choice in patients with rheumatic disease, but an opioid medication should be considered-as well as in patients with intractable pain caused by another disease when alternative therapeutic measures have failed. The principles of opioid medication in rheumatic pain are similar to those in patients with cancer pain. PMID- 18415195 TI - [Painful thoracic spinous processes-a quantative measure of pain?]. AB - A study was conduced in 100 migraine patients and 100 control subjects who did not suffer from headaches to test Dung's hypothesis that the number of painful spinal processes in the throacic spine can be used as a quantitative measure of the degree of pain in the patient concerned. The results show significant differences in the frequency of painful spinal processes between the two study groups. This supports Dung's hypotheses. As it is easy to determine the presence of painful spinal processes the frequency of this phenomenon, the reasons for it and its significance should be the sujbects of further examination. PMID- 18415196 TI - [Pain concept modification in chronic pain patients by an educational video.]. AB - The aim of the study was to evaluate an educational video designed to modify the pain concept of chronic pain patients. It is commonly described that chronic pain patients foster an illness model which is dominated by purely medical assumptions about causes of pain and its modulation and treatment. Furthermore the mostly unrealistic hope for total pain relief which is expected from the pain expert guides the patients' seek for help. These concepts are most probable very dysfunctional in a multidisciplinary pain management setting, where psychological interventions are important elements of the treatment. Therefore the video was designed to initiate changes in the patient's attitude towards a multimodal model of pain. The educational pain video was evaluated in an experimental design using a control group to whom a different video on a health related topic (nutrition) was presented. The Ss participating in the study were 47 chronic pain patients of a pain ambulance and 42 patients of a pain clinic (inpatient setting). The results showed that after viewing the pain video the groups differed significantly in their pain concept as predicted. The use of an educational video, like the one evaluated, seems useful to initiate first steps in illness concept modification by expanding and enriching the patients attitude by assumptions about the influence of psychological factors on pain maintenance and management and shaping realistic attitudes towards treatment. PMID- 18415197 TI - [Trauma-triggered migraine attacks. Review of the literature.]. AB - In children and young adults migraine attacks can be triggered by mild head injury. The literature on this syndrome was surveyed and 50 case reports found to meet the latest criteria of classification requiring at least two similar attacks for diagnosis of migraine (except for common migraine which was excluded from review). 33 subjects had at least one trauma-triggered attack and one identical or similar spontaneous attack, 17 cases at least two similar or identical trauma triggered attacks. An analysis of all cases showed the following features: The symptoms of migraine mostly start with a latency between one and thirty minutes after the injury and dissolve within one day. First attacks without mention of headache were mainly found in children younger than 8 years. Trauma-triggered migraine attacks are well documented for familial hemiplegic migraine, migraine attacks with hemispheric symptoms and attacks with disturbances of consciousness, while the view that posttraumatic transient cortical blindness and transient global amnesia are migraine attacks is insufficiently supported. A hereditary predisposition for a traumatic trigger mechanism seems to be present at least in familial hemiplegic migraine. Nosologic relations to syndromes of secondary neurological deterioration after mild head injury in childhood are discussed. PMID- 18415198 TI - [Not Available]. PMID- 18415199 TI - [Not Available]. PMID- 18415200 TI - [Not Available]. PMID- 18415201 TI - [Not Available]. PMID- 18415202 TI - [Not Available]. PMID- 18415203 TI - [Not Available]. PMID- 18415204 TI - [VIth world Congress on pain 1.-7. April 1990 Adelaide, Axstralien.]. PMID- 18415205 TI - [Not Available]. PMID- 18415206 TI - [Not Available]. PMID- 18415207 TI - [Prevention of chronic low-back pain. Article from the psychological point of view.]. AB - Treatment of chronic low back pain (CLBP) is not only expensive, but is frequently not totally effective. For these reasons, it is important that the risk factors that correlate with the development of chronic pain be considered at the early stage of acute low-back pain (ALBP) in order to implement early treatment to prevent the condition from becoming chronic. Although most of the studies in this field suffer from methodological problems, it appears that the ALBP patients who carry the highest risk of becoming chronic are those who report previous pain episodes, who describe their pain as extending over a wide area of their body and as deep and central, who display no objective findings, who suffer from anxiety and depression, and who "always feel sick." The risk in these patients is further increased when they have a lower educational level and are physically inactive. The recommended early treatment includes shorter periods of bed rest, time-contingent rather than pain-contingent medication, aerobic exericises, education in body mechanics, and behavioural training. In light of the need to contain costs, a program for the prevention of chronic back pain can only be provided for those ALBP patients with an increased risk of having CLBP. Further research on the prevention on CLBP is needed. PMID- 18415208 TI - [Headache in children.]. AB - Idiopathic headaches are probably the most common problem in schoolchildren. The prevalence increases up to 70% in 14-year-old adolescents, with migraines ranging from 10% to 20%. Tension headaches are often understandable on a psychosocial and behavioral basis; in migraine, however, a familial disposition of 60%-80% is well known. Migraine in childhood is initially more common in boys; after puberty girls predominate. At the beginning of migraine development the headache is often not hemisymptomatic in children. Attack duration is shorter than in adults and often lasts less than 4 h. Irreversible neurological deficits are extremely seldom and only occur as ophthalmoplegic attacks in infants. In general, diagnostic studies in headache patients can be restrcted to a neurological examination and electroencephalography. Acute treatment of migraine should include environmental isolation and analgesics such as acetylsalicylic acid or paracetamol; sometimes antiemetic drugs may be useful before. In refractory cases one should try ergotamine. Only a few controlled studies on the prophylaxis of migraine in children have been reported. The first substance introduced in the prophylactic regime was the beta-blocker propranolol. According to results in adults, further optimization could be expected by using thebeta (1)-specific agent metoprolol. New investigations have shown significant efficacy of the calcium antagonist flunarizine as well as low-dose acetylsalicylic acid. Dihydroergotamine drops, often used in pediatric practice, have not proved superior to placebo; the lack of side effects may, however, allow use of the substance as a first-line strategy in staged prophylaxis. In case of failure, nonpharmacological methods such as acupuncture may be tried, especially in older and cooperative children. Vasoconstriction training has been studied in some children; other biofeedback methods and behavioral strategies including relaxation and hypnosis have proved as effective. Follow-up of spontaneous migraine courses in children has shown, that 60% are still or again suffering from migraine as adults. It is presently unknown, whether pharmacological prophylaxis during several months followed by attack-free periods of months or years may influence the long-term prognosis. For tension headaches, transcutaneous electrical nerve stimulation (TENS) has produced good improvement in recent investigations in about 3/4 of cases; full remissions were possible in most cases. A combination of relaxation and behavioral therapy should be recommended where possible, especially in cases of therapeutic resistance. PMID- 18415209 TI - [Oral opioids in patients with non-malignant pain.]. AB - Opioids are given for acute intra- and postope-rative pain relief or for chronic cancer pain. In the literature there are only rare and contradictory reports on the oral administration of opioids for chronic non-malignant pain. However, there is no reason to withhold strong analgesics for patients with severe pain. When all other thrapeutic measures fail to control pain, patients with non-malignant pain can also be treated by opioids. We report 70 patients with severe pain who were given opioids as the ultima ratio in pain therapy: 50 received buprenorphine sublingual tablets, 13 received morphine sustained release tablets and the remaining 7 were treated with other opioids. The mean daily dose was 1.45 mg buprenorphine or 87.6 mg morphine. The dosage increased in 12 of the 50 patients treated with buprenorphine while 5 of the 13 morphine patients needed increasing dosage. The other patients had a constant dosage after the initial period of dose finding. In more than 50% the pain could be effectively controlled by oral opioids. The general performance status (Karnofsky) increased from 63.6% to 74.1%. The typical side effects were constipation and nausea. Prophylaxis of constipation is most important during opioid therapy. No case of respiratory depression or opioid addiction was registered. Our results show that patients with musculo-skeletal and deafferentation pain respond better to opioids than patients with headache. Negative results were observed in some patients with neuropathic pain. The results of the study show that opioids are justifiable for the treatment of non-malignant pain and can be given without danger over a long period of time. Side effects are controlled by additional medication. The principle of opioid administration is prophylaxis of pain -therefore, they should be given "by the clock". Opioids are not only indicated in malignant illness, but also according to severity of pain and by the failure of other measures to control pain. PMID- 18415210 TI - [Pain relief in the final stage of cancer.]. AB - Most patients with very advanced cancer suffer from severe pain, and many studies have demonstrated how this pain can be sufficiently controlled. It is of great importance to find out if the findings are also true during the final stage of cancer and how the treatment must be adapted. We therefore examined the methods and efficacy of providing pain relief for dying cancer patients. This study included 160 patients with cancer in different sites. The pain treatment and pain severity during the last few days and hours of their lives are described and discussed. Analgesic drugs administered orally in 53% and parenterally in 39% of the patients were the mainstay of therapy. Non-opioid analgesics alone were effective in 10% and in combination with weak opioids in 15% of the patients. In 68% strong opioids were necessary to achieve sufficient pain reduction. Morphine was the most frequently used opioid for 96 patients. Oral doses of morphine were 86+/-60 mg/day (15-240 mg/day), and parenteral doses 89+/-74 mg/day (15-360 mg/d). Additional adjuvant drugs to treat specific types of pain or other symptoms of cancer disease were described for 80% of the patients. Non pharmacological measures, such as radiation, nerve blocks or neurosurgery, were of no real importance. Only 4% of the patients treated in the way described experienced severe pain during the final stage of cancer. Systemic administration of drugs is very effective in relieving pain in dying patients. No signs of tolerance to opioids could be observed, even in patients who had been taking opioids for a longer period of time (average 39 days). PMID- 18415212 TI - [Affectivity, irrational attitudes, and pain in patients with rheumatoid arthritis.]. AB - The relationship of patients' pain with emotions and irrational attitudes were reported. The subjects were 128 patients with rheumatoid arthritis (RA). The assessment instruments were the Situation-Reaction Questionnaire (SRQ) and the Irrational Attitudes Questionnaire (IAQ). Pain experience was measured by a pain attribute scale and a visual analog scale, and reported pain behavior by two scales for assessing avoidance and activity in pain situations. The medical control variables were morning stiffness and two indexes of process activity and joint inflammation. Hierarchical regression analyses showed (after the inclusion of medical variables) that affect scales (depression, anxiety, aggression) contribute significantly to the explanation of the variation of pain experience (adjective scale: 3%) and reported pain behavior (avoidance: 11%, activity: 6%). When disease activity and emotions were held constant, the IAQ explained a further 11-14% of pain experience and pain behavior. On the other hand, when medical variables and irrational attitudes were controlled, emotions showed no common variation to pain. According to our results, cognitive concepts seem to be more powerful for explaining pain experience and pain behavior than affective constructions. Implications for the study and the practice of psychological pain treatment are discussed. PMID- 18415211 TI - [Postoperative pain therapy with piritramide and metamizole. A randomized study in 120 patients with intravenous on-demand analgesia after abdominal surgery.]. AB - In this randomized study, the efficacy of i.v. patient-controlled analgesia (PCA) was determined for the opioid piritramide (a pure mu-receptor agonist) and the antipyretic analgesic metamizole (Dipyrone) in three groups of patients following abdominal surgery. The doses of piritramide were 1.5 mg (40 patients) and 3 mg (40 patients) on demand. In addition, we studied the effect of 71 mg metamizole in combination with on-demand boluses of 1.5 mg piritramide in 40 patients. During PCA we estimated the degree of analgesia (verbal gain rating scale, visual analog scale) and monitored the ventilation, vigilance, and typical drug side effects over a period of 24 h. The individual demand for analgesic drugs varied markedly. The mean consumption of piritramide during the study was 46.5 mg in the group with 1.5 mg per bolus and 68.6 mg in the group with 3.0 mg. The resulting pain relief was satisfactory in both groups. The combination of piritramide 1.5 mg and metamizole 71 mg per bolus resulted in a reduction of mean Piritramide consumption to 44.1 mg, and the pain relief was similar to that produced in the group treated with 3 mg piritramide per bolus. The intensity of typical side effects of opioids and antipyretic analgesics (nausea, vomiting, lowering of respiratory frequency, sweating) was low and always easily controlled. The acceptance by patients, nurses, and physicians of PCA was high. PCA with on demand intravenous injection of the combination of piritramide and metamizole improved the degree of analgesia and concomitantly reduced the opioid dose. PMID- 18415213 TI - [Not Available]. PMID- 18415214 TI - [Not Available]. PMID- 18415216 TI - [Not Available]. PMID- 18415217 TI - [Not Available]. PMID- 18415215 TI - [Calcitonin.]. AB - A central antinociceptive effect of calcitonin has been well established in animal experiments. Owing to the lack of appropriate studies, however, a final judgement cannot be made regarding the value of calcitonin in pain therapy. Positive clinical experiences have been reported in the following cases. (1) In isolated osseous tumor pain and in pain caused by tumorous infiltration of peripheral nerve tissue or acute malignant transverse lesions of the spinal cord (with paraplegia), calcitonin can be a suitable supplement to opiate therapy. (2) In algodystrophy calcitonin can be administered in addition to physical therapy. In severe cases, however, this therapy must be supplemented or replaced by sympathetic blockade. (3) In cases of phantom limb pain calcitonin is particularly effective and can also be administered alone as an analgesic. In refractory cases there is usually stump pain of various causes in addition to phantom limb pain. For pain therapy calcitonin should only be administered intravenously in a daily dosage of 1.5-3 IU/kg body weight. If there is no initial success, treatment should not be continued. Dangerous side-effects have not been reported to date. However, dose-dependent side-effects occur frequently, which the patients often consider very distressing. The disadvantages and the "escape" phenomenon that occur during longterm use restrict the value of calcitonin as an analgesic. PMID- 18415219 TI - [Not Available]. PMID- 18415218 TI - [Not Available]. PMID- 18415220 TI - [Not Available]. PMID- 18415221 TI - [Pain treatment in cancer patients.]. AB - Freedom from cancer pain is one of the four priorities of the WHO Cancer Control Programme. Every day 3.5 million people are suffering from cancer pain, and most do not receive adequate relief. A lack of training in cancer pain management at most nursing and medical schools is the principal reason for this, coupled with limited availability of oral strong opioids in many countries. Education is the key to progress in cancer pain management. Health workers must appreciate that: 1. Attention must be paid to all aspects of suffering -physical, psychological, social and spiritual. 2. In advanced cancer, most patients with pain have multiple pain. 3. Pain experienced in carcinoma is not always caused by the tumour. 4. There is more to pain management than the use of analgesics. 5. In the case of opioid-responsive pains, analgesics should be administered by mouth according to a regular time-schedule and with dose increments. 6. Adjuvant medication is generally necessary. 7. Opioid-resistant pains require other forms of treatment. 8. Pain is the physiological antagonist to the central depressant effects of opioids. 9. Opioid tolerance is not a problem in practice. 10. Psychological dependence does not occur in patients receiving opioids for pain relief. 11. Patients receiving analgesics must be carefully monitored. 12. Teamwork is necessary for good results. PMID- 18415222 TI - [Influence of physical therapy on pain.]. AB - The new orientation concerning the therapy of chronic pain has brought about essential progress during recent years. However, physical therapy is often disregarded. Therefore, various possible ways of influencing pain by physical therapy are presented. In contrast to pharmacotherapy, physical therapy is often able to abolish the causes of pain. By means of respiratory control or relaxation it is possible to reduce increased muscular oder vascular reactivity. The unfavourable influences of increased tone of the sympathatic nervous system on the development of chronic pain may be reduced by several methods of physical therapy. The possibility of influencing the nociceptor and the surrounding tissue by electrotherapy is still a hypothesis, but a promising one. Central pain control by physical therapy is investigated most frequently and is of essential importance. It is also of value to take into consideration the psychotherapeutic effects of physical therapy. The integration of physical therapy into a complex treatment schedule may lead to further progress in the treatment of patients with pain. PMID- 18415223 TI - [The applicability of the West Haven-Yale multidimensional pain inventory in German-speaking countries. Data on the reliability and validity of the MPI-D.]. AB - The reliability and validity of a German version (MPI-D) of the West Haven-Yale Multidimensional Pain Inventory (WHYMPI) was assessed in a sample of 185 chronic pain patients. MPI-D shows high internal consistency, valid subscales, and a factor structure that is comparable to the American version. The Interference scale of part 1 includes an additional item and one other item was excluded; the Life Control scale had one item added. In section 2, one item was dropped, and in section 3 only 3 instead of 4 activity scales were found. The questionnaire is sensitive to therapeutic change. The German scale means are lower for the scales indicating more disturbance or severity and higher for the scales indicating less disturbance. It is not clear whether this reduced pain impact is characteristic of German pain patients in general, or whether it is due to the less severely affected sample tested in this study. PMID- 18415224 TI - [Six years' experience with outpatient group treatment of fibrositis patients.]. AB - Since 1982, a total of 88 patients with refractory fibrositis have received treatment as part of an integrated group program within a psychiatric outpatient clinic working closely with the department of rheumatology. The best arrangement proved to be a series of 15 weekly two-hour sessions with groups of 8 patients. The program has three main elements: information, instruction in pain control strategies, and group discussion. The pain control strategies taught are cognitive behavioral techniques, relaxation, and exercises. A controlled study with 27 patients demonstrated that a comprehensive program of this kind was superior to instruction in autogenic training alone. Patient feedback confirmed the importance of the information given and of the group discussions; it is helpful to offer patients a wide range of pain control strategies that they can apply themselves. Our program makes it possible for a greater proportion of patients to learn one or more techniques affording some measure of pain relief. It is important for the therapists to adopt a positive and convincing attitude while restricting themselves to helping the patients to help themselves and avoiding power struggles. They should also be ready to accept the patients and their view of the illness, and not attempt to interpret the illness as the expression of something else. PMID- 18415225 TI - [Results of long-term therapy after lumbar disc syndromes. An analysis of surgically and conservatively treated patients.]. AB - The therapeutic results of operatively and conservatively treated patients with lumbar disc syndromes were reviewed in a retrospective study. The patients were treated during a 10-years period (1976-1985). A total of 330 patients with lumbar disc prolapses were treated in the hospital during this period 44% were treated surgically. The data on 100 operated and 100 conservatively treated cases, registered in this random test sample, have been compared with respect to: pain; neurological deficits; subjective problems and sociomedical questions. The average patient age of both groups was about 41 years, and the patients predominant were male (about 70%). The therapeutic results of both operatively and conservatively treated patients were good, which is also by the high percentage of employment (80%-90%) in the two treatment groups. The critical evaluation showed more neurological disturbances and limited vocational activity in the group of cases operated upon. More than 70% of the operated cases showed radicular syndromes of the follow-up examination although it was not of essential functional importance. The period inability to work and the percentage of disablement were also much higher in this group. The pain symptoms were particularly relevant in our examination. Only 12%-16% of the patients in the two groups that took part in the follow-up examinations reported freedom from pain. It was apparent that atypical pain syndromes were correlated with personality psychological disturbances. Nearly one-third of our patients mentioned psychological problems. The prognosis of the conservative treatment of lumbar disc prolapse was equivalent to operative therapy (disregarding the absolute indications for operations). There were no definite advantages found for either of the two methods of treatment. The necessity for a specialized follow-up treatment of patients with sciatica due to herniated lumbar discs is discussed, and differentiated selection for operative therapy is given. Here the treatment of pain should be considered most important. PMID- 18415226 TI - [Not Available]. PMID- 18415228 TI - [Not Available]. PMID- 18415227 TI - [Not Available]. PMID- 18415230 TI - [Not Available]. PMID- 18415231 TI - [Not Available]. PMID- 18415229 TI - [The estimation of the i.m. morphine-equivalent in cancer pain treatment with different opioids or different routes of administrations. Practical meaning and limitations.]. AB - During the long-term treatment with opioids it is sometimes important to switch the opioid or change the route of administration. The estimation of morphine equivalents can be helpful in this range because it clarifies the dose in milligramm required for different clinical situations. The basis of this estimation is the equianalgesic potency of opioids. One i.m. morphine-equivalent is the analgesic dose of an opioid (i.m. injected) equal to the analgesic effect of 1 mg morphine (i.m.). The relationships between equianalgesic doses and intramuscular and oral routes of applications are listed in tables. The cross tolerance between different opioids during long-term treatment is not complete. To avoid an overdose, we suggest a reduction in the calculated opioid dose of 50%. Additional "rescue doses" can be used during the period immediately the change to provied satisfactory pain control. A new opioid dosage should be calculated every 24 hours based on the basaline dose plus the total quantity of "rescue" medication required by the patient. Useful starting point for calculation an effective dose when changing from one opioid or route of administration to another can result in improved pain control that is more responsive to patient need. The limitations are 1. individual differences in the response to opioids, especially during long-term treatment and in the development of analgesic tolerance, 2. individual differences in the response to alternatives routes of administration, and 3. the unknown degree of cross tolerance among opioid drugs. The scientific meaning of the estimation of i.m. morphine equivalent is discussed. PMID- 18415232 TI - [Editorial.]. PMID- 18415233 TI - [Not Available]. PMID- 18415234 TI - [The "healthy pain patient"]. AB - Despite all of the progress that has been made in diagnostic procedures and the increasing number of treatment facilities available the number of people suffering from chronic pain conditions seems to be growing constantly in all industrialized countries, a fact which is demonstrated impressively by the epidemiology of low back pain. "Chronic" means "life-determining"-chronic pain, as all chronic illnesses represent a turning point in the life situation of the people concerned. They not only affect the patients, but also the members of their immediate social environment. Chronic pain becomes a destructive stigma when society reduces the afflicted persons to the status of the chronically ill. The conceptual framework for health promotion and chronic pain is based on the discussions on lifestyles that have been conducted at WHO/EURO since 1980 and the Ottawa Charter on Health Promotion adopted at the the First International Conference on Health Promotion held in November 1986. The introduction of the concept of the "healthy pain patient" has the goal of raising the competence of the individual and his/her social environment to improve the quality of life in spite of chronic pain. The educational aim is to enable patients with pain to be autonomous and to maximize their potential health. The therapeutic approach is demonstrated by individual case histories. PMID- 18415235 TI - [Psychological models of pain : Critical comments on the Gate Contro Theory and Proposal of a transactional multicomponent process model of pain.]. AB - In clinical pychology, the Gate Control Theory of pain (GCT) is considered a milestone among the psychological approaches to pain. In this paper ten critical issues are raised against the GCT. It is argued that GCT should be abandoned as a basis for the psychological study of pain. GCT is not compatible with numerous neurophysiological results. Furthermore, it claims to provide specific explanations for psychological phenomena that have long been known. Finally, GCT is inappropriate for the analysis of psychological pain processing and the varied consequences of pain. As an alternative to GCT, a comprehensive process model is proposed that incorporates the many consequences of pain. In this "micro-macro model of persistent and intermittent chronic pain", pain is conceptualized as a transactional multicomponent process. First, the basic features of the model are outlined. Second, the methodological consequences that accompany the model are considered. Third, research evidence on the validity of the model is given, in particular with regard to the relationship between pain experience and subjective stress, as well as between pain experience and coping with the pain. Finally, further research questions are formulated. PMID- 18415236 TI - [Intractable cancer pain as a reason for referral : Analysis of pain etiology and previous drug treatment.]. AB - Many studies have demonstrated that cancer pain can be relieved in most cases by suitable analgesic medication. Patients with a diagnosis of "intractable cancer pain", however, are referred to our pain clinic nearly every day. A retrospective study of 1140 patients was therefore performed to evaluate the pain mechanisms and whether analgesic pretreatment had been adequate. Half of the patients (53%) were suffering from pain at more than one site. The most frequent locations were the back (36% of the patients), abdomen (30%), and the thoracic (22%) region. The main pain etiologies were compression or infiltration of pain-sensitive structures by the tumor (84% of the patients), and less frequently oncological treatment (18%), debilitating disease (10%), or causes unrelated to tumor or therapy (9%). Pain could be classified with almost equal frequency as neuropathic, visceral, soft tissue-related, or bone-related. Upon admission to our pain clinic, most patients (86%) indicated pain of severe intensity. The principal causes for the inadequacy of the analgesic pretreatment were: failure to prescribe analgesics (10% of the patients), irregular intake schedule or prolonged intervals between applications (66%), underdosage of nonopioid analgesics (27%) or opioids (42%), and withholding of nonopioid analgesics (30%), strong opioids (14%), or co-analgesic drugs (17%), although their prescription was indicated. The severe pain was thus caused in many patients by simple mistakes in the prescription of analgesics. Terms like "intractable" should be used with caution when referring to cancer pain because they are often unreflected and can make patients and physicians feel helpless or insecure. PMID- 18415237 TI - [Postoperative analgesia with buprenorphine : A comparative study of epidural, patient-controlled intravenous and conventional subcutaneous administration.]. AB - Thirty patients who had undergone elective anterolateral thoracotomy were studied in the surgical intensive care unit to compare the analgesic effectiveness of i.v. self-administered buprenorphine (group A) with that of epidural administration (group B) and of s.c. administration by a nurse of 0.3 mg buprenorphine every 3-4 h (group C, controls). Every 2 h the patients were asked to record their subjektive pain level as a percentage on an analogue scale: zero was to be used for no pain and 100% for the most severe pain they could imagine. the mean of all analogue scores for pain in the first 36 h was 19.4+/-3.1 for group A; 18.4+/-2.3 for group B and 42.0+/-7.4 for group C (P<0.025). When the mean scores were referred to time, it seemed that groups A and B suffered a little more pain immediately after the operation; however, after 4 h the mean scores for these groups were far lower than that for the control group. The amount of buprenorphine used during the first 36 h was 0.036+/-0.006 mg/h per m(2) in group A, 0.021+/-0.002 mg/h per m(2) in group B and 0.038+0.004 mg/h in the control group. The mean interval between two injections was 9.47+/-1.4 h in group A, 2.47+0.5 h in group B, and 5.18+0.7 h in group C. It can be concluded that, at least in patients in an intensive care unit, scheduled administration of pain killers is unacceptable now. The self-administration device has shown the possibility of on-demand i.v. administration, which is no more dangerous than scheduled s.c. administration. Nurses should be instructed to provide analgesic medication on demand. Epidural administration of buprenorphine is superior to self-administration in terms of the amount of drugs used and the dosing intervals. In the quality of analgesia epidural administration and self administration are equal and superior to the control procedure. PMID- 18415238 TI - [What do pain scales measure in patients with rheumatoid arthritis?]. AB - In this study carried out in a sample of 80 patients suffering from rheumatoid arthritis (RA) tried an attempt was made to answer the following questions: 1. are there pain factors with a wider range that are more generally applicable than those covered by current German questionnaires? 2. To what extent can somatic parameters predict pain factors? 3. To what extent can a patient's pain behavior (a patient's activity scores) predict pain factors? The study was based on data collected by means of the Pain Experience Questionnaire (PEQ), the McGill Pain Questionnaire MPQ, the West Haven-Yale Multidimensional Pain Inventory WHYMPI, the Measurement Of Patient Outcome Scale MOPO, as well as six different clinical parameters. By means of factor analysis, two second-order factors were extracted, representing 1. the patient's impairment due to intensive pain and 2. socio emotional consequences of pain. At a statistically significant level, the first factor can be predicted by the clinical variables. Regression of the activity scores on the factor "socio-emotional consequences" suggests a close correlation between the two variables, although the results failed to reach statistical significance. On the whole, the results strongly support the notion of integrating clinical, behavioral and cognitive findings in the diagnostic assessment of chronic rheumatoid pain patients. PMID- 18415239 TI - [Not Available]. PMID- 18415240 TI - [Not Available]. PMID- 18415241 TI - [Not Available]. PMID- 18415242 TI - [Not Available]. PMID- 18415243 TI - [Not Available]. PMID- 18415244 TI - [Not Available]. PMID- 18415246 TI - [Not Available]. PMID- 18415245 TI - [Not Available]. PMID- 18415247 TI - [Not Available]. PMID- 18415248 TI - [Not Available]. PMID- 18415249 TI - [Models of pain development in radicular compression.]. AB - The problem of nociception and pain development in radicular pain syndromes is not clarified. In the pathophysiology of pain of radicular compression caused by stenosis or disc prolapse, morphological complex nerve root/ganglion is the key structure. Chronic compression forces on the nerve structure cause structural changes. Structural deterioration is linked with a change in the electrical membrane properties of the affected nerve root. The membrane threshold shift in nociceptive fibers is an important prerequisite for pain perception in nerve root compression. New biochemical aspects in the pathophysiology of radicular syndromes are presented, which could explain the discrepancy between pain and objective clinical findings. The article concludes that a better understanding of the nerve root pathophysiology will bring a more differentiated pain-management strategy. PMID- 18415250 TI - [The therapy of pain in rheumatic joint-and spine diseases.]. AB - The therapy of pain caused by rheumatic diseases above all must take into consideration the cause of the pain. In rheumatoid arthritis, especially in the early stages, inflammation is the primary cause of the pain. The pain decreases the inflammation subsides following the administration of non-steroidal anti inflammatory drugs (NSAIDs), or corticosteroids, if necessary. The so-called disease modifying anti-rheumatic drugs do not influence the inflammation or consequently, the pain directly, but rather through mechanisms before the local joint process some of which are not exactly known. In later stages of the progressive joint degeneration the NSAIDs only have a limited effect regarding the inhibition of inflammation. In osteoarthrosis, in which the pain is caused by a secondary inflammation and increasingly by capsular, muscular and tendon involvement, the pain is only treated by NSAIDs in active inflammatory stages; otherwise, the treatment is physical activity and medication. In degenerative and static disorders of the spine the pain is caused predominantly by muscular bracing. Therefore, physical and especially gymnastic therapy play a major role. Whether muscle relaxants have an effect on muscle bracing is doubtful. If there is pressure on the ligaments and, in cases of vertebral dislocation with overstraining of the vertebral joints, therapy with local injections is indicated. The pain in osteoporosis is also predominantly muscular and must be treated accordingly. Above all, high doses of calcium and calcitonin are effective analgesics. Moreover, fluoride also acts as an analgesic once the osteoporosis has stabilized. In most cases fibromyalgia, which is mostly of a psychosomatic nature, cannot be influenced by medical therapy. Instead repeated attempts at treatment help to make the affliction chronic with neurotic fixation. Also, as a rule, myotonolytic and tranquilizing substances are not effective. PMID- 18415251 TI - [Chronic pain factor in patients with lumbar disc herniation.]. AB - Using a biopsychosocial model of chronic radicular pain, we conducted a prospective study on the predictability of the therapy outcome in 41 lumbar disc patients from the Department of Neurology, University of Kiel. Before therapy, all patients had an extensive neurological and psychological examination. The criteria for the therapy outcome werepersistent pain and theduration of hospital stay in days. As for the psychological predictors, we examined the amount of depression as a state variable (Beck Depression Inventory BDI), depression as a trait variable (Giessen test), several paincoping modes (Hoppe scale) and the general health locus of control. As somatic predictors, we assessed the duration of pain before treatment, the number of previous operations, motoric paresis and the patient's age. The results indicated that the BDI was the best predictor of persistent pain and of the duration of hospital stay as well. The sensitivity and specificity were more than 90%. Patients with a BDI score >9 remained 8 days longer in the hospital than patients with lower BDI scores. In contrast to this, depression as a personality dimension allowed no correct prediction of patients with persistent pain. Thus, only the situational aspect of a depressive state is a relevant risk factor for chronicity. Overt pain behavior, avoidance behavior and fatalistic control expectations are the best predictors of persistent pain besides the BDI. Patients with persistent pain when discharged from the hospital had significantly more overt pain behavior preoperatively than patients without pain. They admitted that they changed their posture more often; they groaned, grimaced, or rubbed the painful area more often. Thus, these data confirm the operant conditioning theory of Fordyce within a prospective design. Furthermore, patients with strong avoidance behavior in pain situations and with fatalistic health expectations remained 8 to 10 days longer in the hospital. Regarding the somatic factors, only paresis is a significant predictor of these criteria. Patients with clear paresis showed more pain and a longer duration of hospital stay. In general, there was no significant correlation between the organic and psychological predictors, so independent psychological screening and the prospect of psychological interventions are necessary measures to prevent persistent pain in lumbar disc patients. PMID- 18415252 TI - [Prescription of strong opioids by physicians.]. AB - Morphine and other strong opioids are very often needed for the treatment of severe pain. In the FRG the prescription of these analgesics is limited by law, especially for outpatients. Special prescriptions are needed for opioid medication. We analyzed the treatment of outpatients with strong opioids in the first 6 months of 1985 and of 1988. Only 184 members of the AOK Hannover, a large insurance branch with more then 320,000 members, received a prescription of strong opioids from January to July 1985 (=0.057% of all members). On the whole, only 994 prescriptions (=0.094% of the total amount of prescriptions) of strong opioids were issued by the family physicians. During the first 6 months of 1988 we found comparable data: 1,581 prescriptions (=0.145%) for 243 members of the AOK Hannover (=0.075%). In 1985 as well as in 1988 only 16% of the physicians registered in the area of Hannover prescribed strong opioids. Insufficient treatment of outpatients suffering from severe pain is obvious. The complex regulations in the FRG concerning the prescription of strong opioids are the main reason for insufficient treatment. It is necessary to liberalize these regulations in order to treat severe pain better. PMID- 18415253 TI - [Posttraumatic headache : Observations in a pain clinic.]. AB - Following head trauma many different forms of headache can develop. Complaints in the early post-traumatic period appear to be organic in most cases. Later in the course post-traumatic headache frequently shows a psychogenic picture. Describing the causative factors and discussing the therapeutic guidelines, the authors explain how psychological influences can form chronic pain syndromes. PMID- 18415254 TI - [Not Available]. PMID- 18415255 TI - [Not Available]. PMID- 18415256 TI - [Not Available]. PMID- 18415259 TI - [Not Available]. PMID- 18415258 TI - [Not Available]. PMID- 18415257 TI - [Not Available]. PMID- 18415260 TI - [Not Available]. PMID- 18415261 TI - [Pain and pain therapy in children.]. AB - Special aspects of pain in children are pointed out, and the range of relevant conditions and their frequency are compared with adult pain problems. Headache and especially migraine, with incidences of up to 50% and 10%, respectively, are the most common painful conditions to be treated in the pediatric age group. Many less frequent chronic painful disorders claim the need of special, mostly neuropediatric, knowledge; optimal diagnostic precision is important for good therapeutic results. Cancer pain in children is an area of intense interest, with attempts to devies strategies of pain prophylaxis and consistent analgesic treatment. Another problem is that of pain assessment in infants and children; some new diagnostic tools, such as color scales and the Smiley analog scale, can help to solve these specific difficulties. Furthermore, the prediatrician accepts responsibility for the patients' future as adults and is therefore involved in the early prophylaxis of analgesic abuse. The actual and future work of pediatric pain clinics, the conditions in such institutions and the philosophy behind them are presented, and the necessity for research is pointed out. PMID- 18415262 TI - [Pain syndromes affecting the locomotor apparatus. Clinical signs, differential diagnosis and therapy.]. AB - Clinical pain syndromes affecting the locomotor apparatus can become apparent not only in the form of nociceptive pain in the skeleto-mechanical system, but also as neurogenic pain emanating from lesions in the peripheral nerves or nerve roots or as referred pain resulting from disorders of visceral organs. The anatomical structure and basic innervation can contribute to the various characteristics of such deep pain. Within the spinal cord, visceral and somatic afferent fibres converge on nociceptive pathways, resulting in a uniform clinical pain syndrome. The differential diagnosis can be extremely difficult when attempts are made to evaluate its clinical relevance. Pain radiating from deep somatic tissues into the extremities is called "pseudoradicular" pain. Examples are encountered in the "facet syndrome" or the "myofascial pain syndrome". Various types of treatment are available for musculo-skeletal disorders, including physiotherapy, transcutaneous nerve stimulation (TENS) and trigger point infiltration. In clinical practice, however, drug therapy with peripheral analgesics and anti inflammatory drugs is the form of therapy most often prescribed. There is no doubt that these drugs can help a great deal. However, they have only a symptomatic effect without any influence on the origin or course of the disease. Therefore, a critical evaluation of their benefits and risks is required before treatment with such drugs is instituted. PMID- 18415263 TI - [Application procedures and dosage recommendations for postoperative analgesia.]. AB - Systemic application of analgesics is still the most frequently used method of postoperative relief of pain. However, neither intermittent intramuscular nor intermittent intravenous application can provide the patient with a continuous level of analgesia. Lipid-soluble analgesics or those with polar binding that are rapidly metabolized demonstrate an rapid effectiveness. If the analgesia must be administered over a long period, it is due to a low level of lipid solubility, high receptor affinity and low elimination rates. Oral as well as sublingual buccal and rectal applications are characterized by uncertain absorption conditions. There are few investigations on the subcutaneous application of analgesics. After intramuscular administration analgesic levels are achieved within 15 to 60 min, but various conditions may alter the absorption criteria. Intradeltoidal application is preferable to intragluteal injection. Analgesics may be administered intravenously as a bolus, as continuous infusion, or as patient-controlled analgesia. The bolus injection is characterized by a short period of action and the necessity to administer several bolus injections by repeated administration. The continuous infusion of analgesics should begin with the administration of an initial bolus injection. Infusion analgesia should be performed under careful monitoring conditions. The most promising method of pain relief is patient-controlled analgesia (PCA). After an initial bolus injection, the continuous infusion of an analgesic is guaranteed and may be completed by the patient with several bolus injections. PCA requires careful monitoring. We suggest that a special analgesia team to take care of the patient in special analgesia units might be appropriate in the future. PMID- 18415264 TI - [Course of conservatively treatment herniation of the lumbar disc. Radicular deficits and computed tomography findings.]. AB - In Western industrial countries, low back pain is one of the most frequent causes of illness. Between the 4th and 5th decades of life approximately 80% of adults complain of low back pain, lasting for fairly long periods. About 10% of this population must undergo disc surgery once during life. Several authors have discribed good results for lumbar disc surgery in 55%-86% of their cases. Few control data are available, however, on patients with radiologically proven herniation of the disc regression, who were treated conservatively. Their observations indicate that regression of herniation of a lumbar disc is possible using conservative therapy exclusively. A study was therefore carried out on 43 patients with lumbar disc abnormalities, as demonstrated by spinal computed tomography (CT). Initially, 38 of them showed a herniation and 5 protrusion of the disc plus further neurological deficits and radicular pain syndromes. The subjects were followed up for over 20 months (mean) and monitored by CT in order to check the possibility that the CT findings, neurological deficits, and pain would regress a lengthy period of therapy. The results were the following: Initially, all patients complained of severe low back pain and sciatica, leading to subsequent treatment. At the time of follow-up, 15 of them still reported remittent or chronic low back pain, 9 remittent, and 16 chronic sciatic pain but of much less intensity. Before treatment, 40 patients had neurological deficits, whereas at the time of follow-up, only 24 patients still had deficits. In 2 patients the symptoms had not changed and in 2 others slight deterioration was observed. CT control examinations showed clear regression in the extent of disc herniation in 15 patients, in 18 a moderate decrease, and in 9 cases the CT findings had not changed. A favorable tendency towards regression was observed in disc herniations at the level of L5-S1 and in cases showing sequestration of the disc. Herniations of the disc at higher levels between L4-5 and L3-4 or a lateral herniation, reaching the intervertebral foramen, showed on unfavorable prognosis. PMID- 18415265 TI - [The influence of dexamethasone on pain after lumbar disc surgery. A double-blind study.]. AB - Patients operated because of lumbar disc herniations (104 patients) were included in a randomized double-blind study analyzing the influence of dexamethasone versus placebo on postoperative drug requirements and the pain score on the visual analogue scale. High doses of dexamethasone had been administered: 40 mg i.v. on the night before the operation; 8 mg intraoperatively topical perineural application; 8 mg i.v. in the evening of the day of operation; 2x8 mg i.m. on days 1 and 2 postoperatively; 2x4 mg i.m. on days 3 and 4; 4 mg po on day 5 and 6 postoperatively. A significant decrease in the requirement for analgesics was found in the drug-treated group, particularly male patients, and also an impressive reduction in the lumbar pain score. In conclusion, there was good alleviation of sciatic pain in the dexamethasone-treated group of females during the 1st week after operation, but we found no evidence that the agent tested had an influence on the clinical outcome 1 month following the operation. PMID- 18415266 TI - [Oral and maxillofacial cancer pain therapy with sublingual buprenorphine.]. AB - We evaluated the efficiency of sublingual buprenorphine (Temgesic((R)) sublingual, Boehringer Mannheim GmbH) in a controlled study over a minimum of 4 weeks in 15 patients with extreme cancer pain. The tumors included 8 locoregional carcinoma recurrences in the head and neck region, 4 distant metastases of primary squamous cell carcinoma in the oral and maxillo-facial region, 1 tumor after irradiation, 1 non-Hodgkin's lymphoma, and 1 primarily non-operable carcinoma. Pain intensity was continually documented by means of a visual analogue scale (VAS) by the patients themselves. In 14 of 15 the patients it was possible to reduce pain rapidly to one fifth the original intensity with a comparatively low dosage of 0.60 mg buprenorphine (median) daily. PMID- 18415267 TI - [Not Available]. PMID- 18415268 TI - [Not Available]. PMID- 18415269 TI - [Not Available]. PMID- 18415271 TI - [Not Available]. PMID- 18415270 TI - [Not Available]. PMID- 18415272 TI - [Not Available]. PMID- 18415273 TI - [Not Available]. PMID- 18415275 TI - [Not Available]. PMID- 18415276 TI - [Not Available]. PMID- 18415274 TI - [Not Available]. PMID- 18415277 TI - [Not Available]. PMID- 18415278 TI - [Not Available]. PMID- 18415279 TI - [Not Available]. PMID- 18415280 TI - [Caffeine plus analgesics-a significant combination.]. AB - Since the ban of phenacetin and barbiturates in analgesics, caffeine as a constituent of analgesics has recently more often been blamed for promoting the abuse of analgesics. The available relevant literature was reviewed to perform a benefit/risk analysis. The results were as follows. The capability of caffeine to potentiate analgesia was demonstrated in controlled clinical trials involving approximately 12000 patients. This effect does not mean that there is a stronger analgesic effect, but that the addition of caffeine allows to reduce the dose of analgesics significantly, while maintaining the same analgesic efficacy. A minimum single dose of 50 mg caffeine is required to achieve this potentiation. Furthermore, it was demonstrated in controlled clinical trials in about 2000 patients that the maximum analgesic effect is achieved after 30-45 min with caffeine-containing analgesics, but only after 60-90 min with monotherapy. This must be regarded as a great advantage in terms of cumulative toxicity, because additional or higher doses due to insufficienttimeliness of efficacy are thus avoided. Thus, the analgesic benefit of caffeine as a constituant of analgesic drugs can no longer be denied, even though some studies leave room for criticism. (It should be added that this beneficial effect was investigated only for acute treatment with acetylsalicylic acid plus caffeine or with paracetamol plus caffeine). Toxicity studies with combination drugs indicate that there are no risks beyond the known risks of the individual components. On the contrary, the studies suggest that there is a tendency towards underadditivity; with an equal analgesic efficacy, therefore, the toxicological risk of an analgesic plus caffeine tends to be lower than with the respective analgesic alone. With respect to the frequently postulated promotion of analgesic abuse by caffeine, the results of this review are best summarized by citing the statement of the Federal Drug Review Committee "Neurology/Psychiatry" at the Bundesgesundheitsamt (BGA): "There is no evidence showing that a possible potential for dependence in the case of analgesics, such as acetylsalicylic acid or paracetamol, is increased by caffeine. Even though this could be assumed in theory, based on the available data there is no proof of an additional abuse potential for caffeine in combination with analgesics" [11]. The question of whether combination analgesics containing caffeine ought to be sold only on prescription might better be answered in the negative; in Germany, the percentage of self-medication (i.e., without consulting the doctor) in the consumption of prescription analgesics reaches up to 40% (1). Furthermore one could even argue that by introducing prescription schedule for analgesics containing caffeine a higher consumption of analgesic substance might be induced (to achieve an analgesic efficacy equal to that of combination drugs). This however might cause a higher and unpredictable risk (e.g., gastrointestinal bleeding) with proper use in exchange for of a reduction in the predictable risk with misuse or abuse. This would raise major ethical and legal issues. Minimizing one special risk does not automatically reduce the overall risk. PMID- 18415281 TI - [The psychological aspect of cancer pain.]. AB - The perception and expression of pain are primarily psychological phenomena and are not directly correlated with the intensity of the nociceptive stimulus. They are often influenced by earlier experiences of pain, and also by current expectations and fears. The cancer patient interprets pain as a sign of the continued existence and of the progression of the malignant disease: if the pain can be controlled the patient will take this as an indication that the underlying disease can be cured. Inappropriately treated pain, on the other hand, can initiate a vicious circle leading to really excruciating pain and functional destabilization of the patient. Conversely, the personality and the psychological condition of the patient can have a profound effect on how pain is experienced. Fear, for example, is known to exacerbate pain, and fear is often due to a less than ideal doctor-patient relationship, e.g. one in which the patient does not receive adequate information about the disease. Pain can be interpreted as a message that has to be understood before an adequate therapeutic response is possible. For desperate patients who are socially isolated after a long period of illness, pain may be the only way of communicating their unhappiness to other people and of feeling alive at all. Patients with intractable pain are sometimes given placebo therapy, particularly if the pain is interpreted as "only" psychogenic in character or if traditional methods of treatment have failed. Confrontation with terminally ill patients is an especially difficult and frustrating experience for health professionals. The fact that the disease cannot be cured and that a patient is in constant pain reminds the physician of the limitations of curative medicine. This can trigger defense mechanisms in the physician, which may in turn cause insecurity and fear in the patient. In the course of treatment for pain, cancer patients derive most psychological support from the emotional empathy of the therapist, whose availability for the patient is the most important means of preventing the patient's with-drawal into depression. If cancer pain is accompanied by an emotional, psychic and vegetative imbalance, psychotropic drugs are beneficial. In particular, antidepressants and neuroleptics have become an important component of the treatment of chronic pain in cancer patients. Due consideration of the emotional and motivational status of the patient will make it possible to choose between the different effect profiles of these drugs. However, the use of psychotropic drugs should complement, and cannot replace, empathic care from the physician. PMID- 18415282 TI - [The correspondence between pain stimulus and pain sensation.]. AB - The relationship between painful pressure stimuli and induced pain intensity was examined. For experimental pain stimulation in 26 subjects, randomized local pressure on the middle phalanxes of fingers II-IV was used (pressure area 2.56 mm(2), six different pressure levels, three different pressure times). The induced pain intensity was measured by the category splitting procedure. Our results demonstrate that the pain intensity is proportional to the logarithm of the pressure stimulus. This finding is predicted by Fechner's law. The intraindividual and interindividual deviations are normally distributed. The retest reliability between measurements in the morning and in the evening amount to 0.98. Thus, the procedure can be used to objectify the pain intensity experienced. PMID- 18415283 TI - [Not Available]. PMID- 18415284 TI - [Not Available]. PMID- 18415285 TI - [Not Available]. PMID- 18415286 TI - [Not Available]. PMID- 18415287 TI - [Not Available]. PMID- 18415288 TI - [Not Available]. PMID- 18415289 TI - [Not Available]. PMID- 18415290 TI - [Cancer pain.]. AB - There is no uniform etiology of cancer pain. It is essential to understand the pathogenesis of pain as far as possible before a therapeutic modality can be conceived. The anatomical relation of the painproducing lesion to the site of pain perception should be clear (local, projected and referred pain). The origin of cancer-induced pain is classified as follows: malignant, mostly metastatic bone lesions, compression and infiltration of peripheral nerval structures, expansion in limited spaces, distension of liver, obstruction of blood vessels, obstruction and distension of the intestine, other abdominal or thoracic processes that produce visceral pain, infiltration and ulceration of soft tissue in sensitive areas. There are also pain syndromes caused by cancer therapy: post operative, post-radiation and post-chemotherapy pain. Attention is drawn to the difficulties of pain recording and pain measurement. Psychological and social aspects of cancer patients emphasize the importance of a sufficient pain therapy which is divided into non-drug therapy and drug therapy. Various specialities can contribute therapeutic modalities for the treatment of cancer pain. Surgery, orthopedics, neurosurgery, radiotherapy and others have their specific methods. Anesthesiological methods are mentioned in more detail. The celiac plexus block with alcohol as a simple, safe and efficious procedure should become available to any patient with upper abdominal visceral tumor pain. Attention is drawn to the hospice movement, which is more or less unknown in central Europe. Psychological aspects of cancer patient care are considered. Drug therapy is of greater importance than all other methods. That is the domain of the general practitioner. Commonly used analgesic antipyretics and NSAIDs are listed in Table 1. The principles of opioid therapy follow. Due consideration is given to neuroleptics and antidepressive drugs. Information about hormones (corticosteroids, calcitonin a. o.) in cancer pain therapy conclude this survey. Enormous differences of morphine use (Austria: 0.66 kg vs Denmark 16.59 kg per million people per year) indicate that there is a great demand for further professional education in this field. PMID- 18415291 TI - [Perceptional disorders in migraine.]. AB - Perceptual disorders in patients with migraine are common and therefore well known in the optic sense. However, a review of the literature reveals a variety of perceptual phenomena with regard to all of the senses as well as disturbances in body image and physical sensations. The criteria for interpreting these phenomena in the context of migraine disorder, even if they are not accompanied by headache, are: the reaction to migraine therapy; previous occurrence, together with a migraine headache; a family history of migraine. From a psychopathological perspective, selected features of the phenomena and questions regarding terminology are discussed: the phenomena should be referred to as hallucinations, the headache itself is real and should not be called a hallucination. PMID- 18415292 TI - [The effect of central nervous modulation on the quality of epidural blockade.]. AB - Conventional clinical dosages of local anesthetics may not be sufficient to block conduction of all afferent impulses, which is important for an effective epidural sensory blockade. Further depression of the transmission of information can be accomplished by central modulating mechanisms, preventing sensory perception. Since there are interindividual differences in the quality of modulating mechanisms, standardized depression of impulse propagation by the local anesthetic is associated with a wide variation in the total depression of afferent impulses. For that reason, the results of a particular dosage of a local anesthetic will be variable regarding the amount of epidural sensory blockade perceived. A continuous epidural technique using a catheter permits the dosage to be individualized by means of additional injections. Patients with a low pain threshold require a more highly concentrated local anesthetic. The addition of opioids to the local anesthetic will permit the use of lower concentrations of the local anesthetic itself. Optimal use of diagnostic, prognostic, and therapeutic epidural blockades is possible only if the influence of modulating mechanisms is taken into account. PMID- 18415293 TI - [Self-control training for patients with chronic pain.]. AB - Psychological methods for the treatment of chronic pain have been developed within the contextual framework of behavioral medicine during the past few years. Some of these self-control approaches focus on the patient's capacity to cope actively with pain. Standardized self-control training for outpatient group treatment for patients with chronic pain is described. The training consists of training in progressive muscle relaxation and pain-control techniques such as attention diversion, imagination, and relabeling of sensations as well as modification of pain-related attitutes and awareness. Thirty-three patients with chronic back pain were assigned either to one of four treatment groups (n=22) or to a waiting list control group (n=11). After 12 sessions of group treatment, patients showed significant improvement regarding daily ratings of pain intensity and mood and a significant decrease in depression and general physical complaints other than pain; the waiting list patients remained the same. Results showed good maintenance at the half-year follow-up. PMID- 18415294 TI - [The difficulties involved in arriving at a comprehensive definition of the phenomenon pain.]. AB - Although pain is part of our everyday life, to date there has been no generally accepted definition of pain. In this article is presented on a survey the various attempts to define pain. Four types of definitions can be distinguished: (a) definitions of pain oriented to a stimulus; (b) definitions of pain oriented to a reaction; (c) definitions of pain oriented to the experience of pain; (d) definitions of pain by determining the components of pain. The advantages and disadvantages of these types of definition are discussed. Using a component model, it is demonstrated that scientific investigation of pain can only grasp single aspects. Therefore, the nature of the subject implies that interdisciplinary cooperation is necessary. PMID- 18415295 TI - [Not Available]. PMID- 18415296 TI - [Not Available]. PMID- 18415297 TI - [Not Available]. PMID- 18415298 TI - [Not Available]. PMID- 18415299 TI - [Not Available]. PMID- 18415300 TI - [Not Available]. PMID- 18415301 TI - [Not Available]. PMID- 18415302 TI - [Not Available]. PMID- 18415303 TI - [Not Available]. PMID- 18415304 TI - [Not Available]. PMID- 18415305 TI - Opioid agonists: Modes of application and dosing regimens. PMID- 18415306 TI - Clinical pharmacology of opiate agonist/antagonist (OAA). PMID- 18415307 TI - Clinical use of peripherally acting analgesics (PAA). PMID- 18415308 TI - [Not Available]. PMID- 18415309 TI - [Not Available]. PMID- 18415310 TI - [Not Available]. PMID- 18415312 TI - Clinical pharmacology of spinal opioids. PMID- 18415311 TI - [Not Available]. PMID- 18415314 TI - [Not Available]. PMID- 18415313 TI - [Not Available]. PMID- 18415315 TI - [Not Available]. PMID- 18415316 TI - [Not Available]. PMID- 18415317 TI - [Not Available]. PMID- 18415319 TI - Pain management after outpatient surgery. PMID- 18415318 TI - [Not Available]. PMID- 18415320 TI - [Not Available]. PMID- 18415321 TI - [Not Available]. PMID- 18415322 TI - [Not Available]. PMID- 18415323 TI - [Pathophysiology of ischemic cardiac pain.]. AB - Cardiac pain is a conscious experience that can be explored only indirectly with experimental approaches. The exact machanisms eliciting cardiac pain still remain obscure. The afferent fibres running in the cardiac sympathetic nerves are regarded as the essential pathway for the transmission of cardiac pain. Atria and ventricle are abundantly supplied with sympathetic sensory innervation. In the spinal cord, impulses transmitted by the sympathetic pathway converge with impulses from somatic thoracic structures onto the same ascending spinothalamic neuron which probably explains the mechanism of referred pain (=projection of pain to another organ). Two hypotheses have been put forward to explain the peripheral mechanism for nociception. The intensity mechanism assumes that pain results from an excessive stimulation of receptive structures normally stimulated at lower levels whereas a specific sensation is considered to result from an excitation of a well defined nociceptive apparatus. Ventricular sympathetic afferent fibres whether myelinated or unmyelinated, always possess some mechanosensitivity and respond to normal chemical and mechanical stimuli, thus displaying properties of polymodal receptors. Afferent vagal fibres may contribute to the mechanisms of cardiac nociception by modulating the threshold and characteristics of pain. Experimental studies identified three main mechanisms, which may be responsible for eliciting cardiac pain during ischemic periods in humans: a) nonphysiological motion of the ischemic left ventricular wall (bulging) and an excitation of mechanical receptors by passive stretching. b) The excitation of free sensory nerve endings by chemicals such as bradykinin, PGE(2), adenosin, histamin or potassium. c) A combination of a and b: algogenic chemicals may sensitize mechanical receptors and therefore lower their threshold for nociception. PMID- 18415324 TI - [Development and therapy of the pain syndrome of reflex sympathetic dystrophy. Clinical expression, experimental investigations, and new pathophysiological considerations.]. AB - Reflex sympathetic dystrophy (RSD) is a disease of the extremities that can be elicited by different factors, occurring at different sites (e.g., trauma, herpes zoster, myocardial infarction). Independently of its etiology, however, the clinical symptoms of RSD are found most often in distal parts of the extremities affected (hand or foot). In a generalized distribution pattern, the following signs, representing a triad of autonomic, motoric and sensory disturbances, are commonly observed in these regions: 1. dysregulation of blood flow to the skin and of sweating, together with diffuse swelling, 2. impairment of movement and muscular strength; 3. diffuse sensory skin disturbances and spontaneous pain of ariable character (e.g., burning, throbbing, aching, shooting). Pain sensation is generally diffuse; in most cases it is deep and less often, superficial (probably representing bone or skin pain, respectively). This triad occurs at the very onset of RSD. If the distribution pattern is generalized, it can be used as a diagnostic criterion for RSD. Our experimental results support the idea of disturbances of skin blood flow related to abnormal vasoconstrictor outflow. This assumption is primarily based on two observations: 1. 73% of 97 RSD patients (upper extremity affected) showed systematic side differences in fingertip temperatures at room temperature. All points measured on the affected side had higher (n=51) or lower (n= 20) temperature values than corresponding sites on the healthy extremity. Such systematic side differences were found only in 16% out of 79 healthy subjects (p-0.90). Significant intercorrelations between the results of the application of mechanical pressure and ischemia support the validity of these two stimuli. Stimulation by cold, on the other hand, did not reveal such coherence. Repeated mechanical stimulation caused sensitization that did not occur with ischemia or cold. There were no significant differences whether the experiments were performed in the morning or in the evening. Experimental pain induction either by mechanical pressure or by ischemia judged in accordance with the category subdividing procedure produce reliable and valid results; the methods are easily applicable and economic. PMID- 18415366 TI - [Not Available]. PMID- 18415365 TI - [Not Available]. PMID- 18415367 TI - [Not Available]. PMID- 18415368 TI - [Not Available]. PMID- 18415370 TI - [Not Available]. PMID- 18415369 TI - [Not Available]. PMID- 18415371 TI - [Not Available]. PMID- 18415372 TI - [Clinical symptoms and classification of migraine.]. AB - Since the beginning of the history of man migraine has been described in all civilizations. It would therefore seem helpful if migraine were to be classified, for diagnostic reasons and in order to study the scientific hypotheses, according to the manifold clinical symptoms, as well as the lack of typical pathophysiological, morphological and biochemical findings. In this review the new classification of headache forms according the the International Headache Society (1988) is presented in comparison to earlier classifications, with special attention to migraine. On the basis of this classification, the individual forms of migraine are described from the viewpoint of the clinical symptoms. The classification of the International Headache Society represents a good basis for reclassifying headaches, which is still under discussion. PMID- 18415373 TI - [Clinical aspects of pathophysiological mechanisms in migraine.]. AB - Migraine is more than the pain involved in the "migraine attack." Before the onset of pain many clinical symptoms can be observed. These symptoms may be classified as vegetative, affective, and vascular. Brain perfusion is altered during migraine attacks as well as during the intervals between attacks. These "more recent" findings are important because brain perfusion is controlled by metabolic and by neurotransmission mediated pathways: 750 ml blood/min is available in brain perfusion. The skull, on the other hand, limits the volume of blood in the brain to 130 ml. Control of the shift of blood inside the brain, with a chance of maximal blood flow or strictly limited blood volume, may be one of the most important problems in neurotransmission mediated cerebral perfusion control. The most important neurotransmission systems of cerebral perfusion control are those that are believed to be involved in affective and vegetative symptoms. It must be assumed that platelets are involved in migraine. Platelet reactivity is enhanced in migraine patients during the interval between attacks. When a migraine attack occurs a release of platelet serotonin and a further increase of platelet reactivity can be observed. Platelet activation in these cases is comparable to the situation in transient ischemic attacks. During transient ischemic attacks, platelet serotonin has been found to be enhanced in the area of transient ischemia. Serotonin is a neurotransmitter, low concentrations of which induce vasodilation, while higher concentrations induce vasoconstriction. It may be assumed that platelet serotonin is a potent vasoregulating substance that may interact in the brain vessels with the neurotransmission controlled perfusion. The hypothesis of an (inborn) instability of the interaction of cerebral neurotransmission systems in patients suffering from migraine is in accordance with the vegetative and affective symptoms in migraine, the observed imbalance of neurotransmission mediated cerebrovascular autoregulation and the irritation of platelets in migraine attacks, as well as in the interval between attacks. The "modern" treatments of migraine with acetylsalicylic acid, ergotamin and/or beta blockers are discussed in relation to this proposed hypothesis of a migraine pathophysiology. PMID- 18415374 TI - [Migraine and stress. Psychobiological investigations of a diathesis-stress model of migraine.]. AB - Pathophysiological models of migraine describe stress as a potential activator of migraine. Clinical observations have shown that stress over prolonged periods leads to migraine, particularly in the relaxation period (e.g. sleep) that follows the stress situations. However, psychophysiological investigations have not been able to prove a direct correlation between stress and the reaction of peripheral and central vascular mechanisms. For this reason, a response specificity in migraine has not been postulated to date. The present paper discusses the relationship between stress, physiological and biochemical processes with regard to a diathesis-stress model of migraine. In an experiment, 17 migraine patients were compared with 16 controls with regard to reactivity of the peripheral resistance of the temporal artery during extremely aversive stress. The peripheral resistance was measured by Doppler ultrasonography. The results showed a significant response specificity in migraine patients with respect to peripheral resistance. The results are discussed using a diathesis stress model of migraine. PMID- 18415375 TI - [Coping with stress and pain in migraine patients.]. AB - During a semi-structured interview 82 migraine patients were asked biographical and illnessrelated questions. They completed psychological instruments on coping behavior (Stressverarbeitungsfragebogen), self-concept (Frankfurter Selbstkonzeptskalen), attributional style (IE-SV-F), illness behavior, and illness-related attributions (Tubinger Attributions-fragebogen). The theoretical background of this research is a cognitive model of coping with stress and illness. The results support the interrelations between coping with stress and coping with illness assumed in the model. They justify considering coping with illness to be scopespecific coping behavior. Furthermore, the results emphasize the importance of cognitive processes for stress-coping in general, as well as for illness-related coping behavior. With regard to personality variables, migraineurs, as compared with healthy persons, show to a larger extent coping strategies that are apt to maintain rather than to reduce stress, such as resignation, withdrawal, and avoidance behavior. They also have a more unfavorable selfconcept of achievement, emotional stability and selfassertiveness, lower self-esteem and a more external pattern of causal attributions. Some of the pain behavior strategies could be identified as being focused on illness (guarding behavior, avoidance and social withdrawal, resignation and complaint); only the attempt to relax is regarded as being focused on health. Migraine patients show a preference neither for medical nor psychological causal attributions of their illness but score significantly higher on medical than psychological control attributions. The results have implications for psychological therapy. PMID- 18415376 TI - [Studies on the predictive factors of treatment success in combination headache.]. AB - Sixty-five patients (51 women, 14 men), who were 41.9 years on average, suffered from combination headache (migraine and tension headache) and were under therapy with propranolol and amitriptyline-N-oxide. The following predictive factors were tested with regard to the success of therapy: age, sex, duration of the migraine and tension headache illness, and frequency of use of analgesic and ergotamine preparations. The prerequisite was a statistically valid classification of responders and non-responders. Responders were defined as patients with at least 50% reduction in their symptomatic parameters (duration and frequency of migraine attacks or tension headaches between the beginning (t(0)) and end of the 3-month treatment (t(3)). Both the migraine treatment and the tension headache were standardized and headache records were kept. The statistical procedure used permitted clear differentiation between responders and non-responders, but these predictive factors did not make it possible to distinguish a subgroup of potential responders. PMID- 18415378 TI - [Not Available]. PMID- 18415377 TI - [Algesimetric experiment on pain sensitivity in the pericranial musculature. Circadian rhythms and corresponding psychic variables.]. AB - Many types of headache that occur in the form of attacks show a notably high incidence at certain times of the day. Attacks are often accompanied by emotional instability, irritability, exhaustion and other impairments of well-being. The cause for the frequent daytime occurrence of attacks is not clear. One possible explanation could be the variability in a person's sensitivity to headache over the course of the day. For this reason we set out to investigate whether pain sensitivity to experimentally-induced headache varies in a circadian manner and whether this variation correlates to changes in the current state of well-being over the course of the day. In 24 healthy, voluntary subjects, headache was induced experimentally at 0200, 0600, 1000, 1400, 1800, and 2200 hours. Blood criculation in both superficial temporal and occipital arteries was reduced by applying a cuff to the head and inflating it to 200 mmHg, thereby producing a continuously increasing, bilateral, dull, frontal headache. The subjects scaled the intensity continually, using the category subdividing procedure ranging from pain threshold to pain tolerance level. In addition, 17 dimensions of the current state of well-being and 12 personality traits were recorded at the above mentioned times in a standardized quantitative manner. At low levels of headache intensity there were no significant differences in pain sensitivity. Sensitivity to very intense headache, however, varied significantly over the course of the day: sensitivity was greatest at 0200 hours; it decreased at a constant rate until 1400 hours, and increased again continuously until 2200 hours (P/=150 mug epidurally. Clonidine is effective in reducing acute and chronic pain. In combination with opioids the action of the opioids is intensified. Clonidine intensifies and prolongs the action of local anesthetics. If opioid tolerance occurs, epidural clonidine alone or in combination with an opioid has good antinociceptive action.Midazolam, a water-soluble benzodiazepine, was injected spinally for the reduction of pain for various indications (postoperative, malignancy, chronic back pain, spinal spasticity). Spinal benzodiazepine should not be injected into the spine in patients until it has been proven that there are no neurotoxic effects. Intrathecally injectedbaclofen is a well-known means of reducing spinal spasticity. Used in this way, it may have a secondary analgesic effect. No significant direct analgesic effect has so far been demonstrated. Spinalcalcitonin often leads to insufficient pain relief when given alone. Combination with an opioid may reduce the dosage of the opioid. Nausea and vomiting are frequent side effects of spinal calcitonin. Intrathecalsomatostatin produces antinociception. However, in animal studies neurotoxic action has been observed. Administration in man has not yet been proved to be safe. Spinalketamine has procluted controversial results in clinical studies, and has not yet been excluded that the substance is not neurotoxic.Lysine acetylsalicylic acid (L-ASA) has been given intrathecally for the therapy of severe cancer pain and chronic back pain. In most patients good analgesia was observed up to 2 months after a single injection. If neurotoxity can be excluded, L-ASA may be an alternative in the therapy of cancer pain before neurodestructive therapy is done. PMID- 18415441 TI - [Specific aspects of perioperative pain relief in children.]. AB - Pain relief in children during the perioperative period can be provided by means of peripherally or centrally acting analgesics or of regional anaesthetic techniques. Narcotics or regional blockde are indicated when peripherally acting analgesics prove inadequate to abolish pain. Side effects of narcotics must be taken into account: opioids must not be administered unless continuous safety monitoring of the child's respiration is assured. If narcotics fail to relieve pain, regional anaesthesia may be indicated. All advantages and drawbacks of the various techniques that might be appropriate must be considered: the technique involving the least risk and side effects is the anaesthetic technique with a broad margin of safety when applied by an anaesthesiologist who has experience with paediatric regional blocks include topical anaesthesia, local infiltration, peripheral nerve blocks (e.g. nervi dorsalis penis, plexus axillaris) and caudal epidural blockade. Caution must be exercised whenever narcotics are administered systemically or epidurally; side effects must not be underestimated, even under conditions of intensive care observation. The provision of effective pain relief is a rewarding task-and particularly in little children. PMID- 18415442 TI - [Modification of the guanethidine block for diagnosing reflex sympathetic dystrophy.]. AB - For diagnosis of the pain origin in reflex sympathetic dystrophy (RSD), sympathetic blocks, including the intravenous regional guanethidine (IVRG) block after Hannington-Kiff, are recommended. Since the results obtained with this kind of block are sometimes unsatisfactory, modifications were made to increase its technical efficacy. PATIENTS AND METHODS: Forty-seven RSD patients were investigated. Among the typical triad of RSD symptoms, all patients showed distally generalized oedema of the affected extremity and complained of spontaneous pain. This pain showed an orthostatic component in all but two patients. The modifications of the IVRG-block were: (1) distal (in contrast to proximal) suprasystolic compression of the affected extremity before i.v. injection of guanethidine (GA); (2) reduction of the recommended dose of GA (10 30 mg) to 2.5 mg GA for each block; (3) no additional local anaesthetics, which are recommended for suppression of the typical pain following GA injection, since this pain allows identification of the distribution of the GA (complete? incomplete?) within the symptomatic area; (4) repeated subsequent injection of physiological saline to improve the distribution of GA within the symptomatic area. RESULTS: Injection of GA and repeated injections of saline (mean: 43 ml, range: 10-95 ml) elicited pain-lessening in intensity with repetition-in the same region as the spontaneous pain of RSD in all but the two patients mentioned above. Following the modified IVRG block, all but these two patients experienced significant pain reduction (Table 1). In total, 87% of all 47 patients under study were acutely (for at least 1 day) pain-free after the block. CONCLUSIONS: The modified IVRG block seems to have a high diagnostic value with respect to the presence of a sympathetic contribution to the pain in RSD. As a reduced GA dose/block is used, this form of the block appears to be safer than other kinds of sympathetic blocks. PMID- 18415443 TI - [Assessing disability in chronic pain patients.]. AB - Despite its importance for chronic pain diseases, pain-related disability is a poorly defined concept with theoretical deficits. The distinction between impairment, disability, and handicap proposed by the WHO is an important contribution to clarification of the disability construct. With reference to four criteria (underlying disability construct, degree of behaviour generalization, assessment mode, scope) different assessment procedures of pain-related disability are presented and the advantages and disadvantages of each are discussed. A multidimensional approach to pain-related disability is advocated. Diagnostic information yielded by different assessment procedures should be viewed as complementary rather than as exclusive data sources. A German version of a self-report instrument (Pain Disability Index) that assesses disability in chronic pain patients is presented. The results from four data sets concerning different aspects of reliability and validity confirm the good psychometric properties of the instrument. The instrument can be used in chronic pain research as well as in clinical contexts. It is recommended that subjective disability data be complemented by behavioral observation and additional data sources (e.g. assessment of disability by the spouse/partner). PMID- 18415444 TI - [Postoperative pain management on surgical wards-organization of an anaesthesiology-based acute pain service.]. AB - There have been major advances in knowledge and efficient techniques for pain relief have been developed during recent years. Nevertheless, many patients on surgical wards still suffer from severe pain following surgery or trauma. Therefore, in the University hospital of Kiel (Germany) an anaesthesiology-based acute pain service (APS) was established in 1985 to improve this situation. ORGANIZATION OF THE APS: The anaesthesiologist in training, who is in overall charge of the recovery unit, serves as an APS for surgical wards and is supervised by a consultant. The anaesthesiologists on duty are on call after regular working hours. The activities of the APS are as follows: induction of adequate postoperative analgesia in the recovery unit for all patients and selection of patients for further special analgesic treatment; clinical rounds including all patients receiving regional analgesia or patient-controlled analgesia (PCA) every morning and throughout the day if necessary; additional consultations on postoperative pain management for other patients on request; assessment and documentation of the clinical status of each patient, the quality of analgesia and side effects; written orders for further treatment (Fig. 1); continuing consultations and informal education for ward nurses, physiotherapists and surgical staff, and formal medical training in postoperative pain management for ward sisters/ charge nurses ACTIVITY OF THE APS: A total of 1947 patients were treated on normal wards from 1985 to 1992. Epidural analgesia was performed using a standard protocol with bupivacaine 0.175-0.25% continuously with top-ups if needed. Since the introduction of an APS, complications of epidural analgesia such as hypotension and insufficient analgesia due to dislocation or other technical complications, have declined significantly in recent years (Fig. 3). Epidural opioids were used in only 46 selected cases on surgical wards. Nevertheless, there were two cases of marked respiratory depression. PCA was performed according to a standard protocol with tramadol or piritramide without background infusion. The loading dose was titrated in the recovery unit. The median duration of PCA was 4 days (range 1-23 days), and the median dose of piritramide was 45 mg/day (range 3-226 mg). There were two cases of somnolence and respiratory depression. CONCLUSIONS: Since the majority of patients used patient-controlled or epidural analgesia for more than 48 h (Fig. 2), it is emphasized again that these techniques should not be restricted to high dependency areas such as ICU or recovery units. The introduction of an APS is an adequate approach towards more efficient and safer pain management on surgical wards. This requires standard protocols, standard monitoring and trained ward nurses. PMID- 18415445 TI - [AIDS and pain management-a survey of German AIDS and pain management units.]. AB - The number of AIDS patients is steadily increasing. According to the literature these patients are often in severe pain. METHODS: We evaluated pain diagnoses and treatments with two almost identical questionnaires for AIDS treatment units (ATU) and pain management units (PMU). Questions dealt with unit type and size, number of patients treated per year and the proportion of intravenous drug users. The units were also asked to give an estimate of pain aetiologies, pain types and localizations and treatment modalities offered. RESULTS: Completed questionnaires were returned by 38 of 235 ATU and 85 of 127 PMU. In the ATU, 16% of the patients (estimated at 580 patients per year) had pain requiring treatment. In 26 of the PMU approximately 120 AIDS patients per year were treated, while 59 PMU had not yet seen any AIDS patients. Pain was caused mainly by opportunistic infections and by neurological syndromes connected with AIDS. Pain aetiologies could not be differentiated in the ATU in 22% of patients (PMU 9%), and pain types in 33% (PMU 9%). Neuropathic pain (ATU 38%, PMU 89%) was more frequent than nociceptive pain (ATU 29%, PMU 36%). The treatment modalities were systemic pharmacotherapy in 76% of ATU and 73% of PMU and nerve blocks in 37% of ATU and 42% of PMU. In 82% of ATU the staff thought their analgesic therapy was adequate, and in 92% staff were interested in closer cooperation with PMU such as was currently practised in only 6 of the 38 units (16%) that responded. CONCLUSIONS: The high incidence of complicated neuropathic pain syndromes in AIDS patients requires a sophisticated therapeutic approach. Closer cooperation between AIDS specialists and pain specialists, comparable to that already existing for other patient groups, is therefore desirable. PMID- 18415446 TI - [Survey on the booklet "Short guidelines for cancer pain-management" issued by the working group "cancer pain" of the german association for the study of pain.]. AB - INTRODUCTION: An interdisciplinary working group on cancer pain was founded by the German Association for the Study of Pain in 1991. The goal of this group is the development and dissemination of training and information material, guidelines and curricula in the fields of cancer pain, palliative medicine and the legal regulations for the prescription of opioids. METHODS: Two editions of short guidelines on cancer pain management have so far been distributed throughout germany (a total of 15 000 copies). In preparation for a revised version, a questionnaire with five questions was sent out to practitioners asking for information on the acceptability of the guidelines and their usefulness in the daily work in practice. RESULTS: Only 167 of 670 (25%) questionnaires were returned. In almost 75% of these, the guidelines were reported to be informative, and in almost 100% as easily comprehensible. For their daily work, 56% of the respondents found the guidelines very helpful and 40% rather helpful. CONCLUSIONS: The results of this survey clarify the need for concrete guidelines in cancer pain management which are at the same time readily understandable. The new edition of 40 000 copies of the "Abbreviated Guidelines for Cancer Pain Management" was published in October 1993. Because of the good response, a further booklet on "Cancer Pain Management in Children" is being planned. PMID- 18415447 TI - [Pain therapy-integration of psychological diagnosis and therapy.]. AB - A multidisciplinary approach, acknowledged as being the best treatment strategy for a wide range of chronic pain patients, requires cooperation at least between trained professionals in the medical and the psychological disciplines. Psychological criteria are formulated to bridge persisting gaps in the interdisciplinary implementation of pain treatment strategies. PMID- 18415448 TI - [Preemptive analgesia: a clinical reality or a scientific hypothesis?]. AB - Numerous experimental studies have shown that systemic or regional administration of analgesics prior to a nociceptive stimulus results in a significant reduction of analgesic requirements compared with the administration of the same analgesic dose given after the nociceptive stimulus. This phenomenon is called "preemptive analgesia". Recently several clinical studies have been conducted to determine whether "preemptive analgesia" also occurs in humans. Regional and systemic opioids, local anesthetics and non-steroidal anti-inflammatory drugs have been investigated so far. Most studies have failed to show a significant reduction in postoperative analgesic requirements with preemptive analgesia. Even in studies with positive results the reduction in analgesic requirements was limited and without clinical relevance. Further studies should focus on the questions which analgesics and which administration routes might provide clinically significant "preemptive analgesia" and how long analgesia should be prolonged into the postoperative period. PMID- 18415449 TI - [Systemic analgesia during labour.]. AB - Systemic analgesia is used in obstetrics to alleviate the pain in labour and to prevent adverse effects on the fetus due to maternal pain and stress and subsequent complications such as prolonged labour. To supplement psychological support tranquillizers such as diazepam are useful in allaying anxiety and increasing patients' acceptance of labour. Possible side-effects include neonatal hypothermia and poor muscle tone of the newborn when large doses are given. When pain is more severe opioids should be administered, since less potent analgesics cannot provide adequate pain relief in obstetrics. The most important side effect of opioids is the dose-related respiratory depression, which can be harmful to the neonate. Therefore, large doses (>150 mg pethidine) should be avoided. However, even moderate doses (100 mg pethidine) may adversely affect infant neurobehaviour. Thus, in many cases adequate pain relief afforded to parturients by systemic analgesia may result in altered adaptive functions of the newborn. This makes it reasonable to consider alternative methods, including epidural anaesthesia, which is highly effective and fairly unproblematic. Drug administration in the management of labour pain can be recommended if only small doses are needed and in parturients who refuse regional anaesthesia or for whom it is contraindicated or not available. PMID- 18415450 TI - [Interpleural analgesia : A topical review.]. AB - Interpleural analgesia is a method of postoperative analgesia that was developed by Kvalheim and Reiestad in 1984. The main indication is postoperative pain after unilateral thoracic and upper abdominal surgery. Many authors report good analgesic effects and better postoperative lung function following cholecystectomy. There is some controversy on the effectiveness of this method after thoracic surgery. Further indications are post-traumatic pain after multiple rib fractures and chronic pain in the upper abdomen (carcinoma of the pancreas, chronic pancreatitis). The local anaesthetic of choice is bupivacaine (in concentrations of 0.25-0.75%, injection volumes of 10-40 ml, with or without epinephrine, applied as bolus or infusion), but others, such as lidocaine or morphine, are also being tested. Risks involved in this method are pneumothorax when the catheter is placed blind and the systemic toxicity of the local anaesthetic. This review provides information on the mechanism of action, the technique, the clinical use to date and possible risks. PMID- 18415451 TI - [Pre-emptive analgesia-prophylaxis of postoperative pain.]. AB - Pain prophylaxis is an everyday experience in clinical anaesthesia. There is now considerable experimental evidence that short-term nociceptive stimuli evoke a long-lasting excitatory state of the central nervous system. This excitatory state can be largely prevented by relatively small doses of anaesthetics (local anaesthetics, opioids) given prelesionally. Suchpre-emptive analgesia is the theoretical basis of the clinical experience cited above. This experimental knowledge has clinical applications in the field of postoperative analgesia which, as is well known, is the Cinderella of anaesthesia. Al-thoughs sound knowledge of the methods is available, postoperative pain relief is very often inadequate. because of organizational difficulties. Pre-emptive analgesia reduces the frequency and intensity of postoperative pain. Infiltration of the surgical incision with local anaesthetics, regional blockades, and spinal and epidural analgesia, all performed preoperatively, and also the administration of analgesies with the premedication, produce postoperative analgesia lasting longer than the known duration of any of these alone. The efficiency of these methods has been scientifically proved during recent years. Pre-emptive analgesia is advantageous in out-patient surgery as well as for routine clinical anaesthesia, and has proved effective in the prevention of phantom limb pain. Many questions on the nature and clinial application of pre-emptive analgesia are still unanswered. However, its ease of performance and the clear clinical advantages of pain prophylaxis mean that it should have a place in the everyday practice of anaesthesia. PMID- 18415453 TI - [Acupuncture treatment for the relief of gonarthrosis pain-a controlled clinical trial.]. AB - The analgesic effect of acupuncture in chronic gonarthrosis pain was studied in a placebocontrolled trial completed by 97 patients. Each patient was treated twice a week, receiving 10 acupuncture treatments in all. Before and after tee course of treatment all patients were examined by an unbiased independent examiner and the overall pain score was measured over 10 days using VAS scales; functional parameters (resilience) were measured with a modified Lysholm questionnaire. Patients in the verum group (n=71) were treated according to generally accepted acupuncture treatment recommendations. Patients in the placebo group (n=26) were treated with sham acupuncture at non-acupuncture points on the homolateral leg. A follow-up examination was carried out after 3 months. After ten treatments the overall reduction in pain score was 47.5% in the verum group (follow-up 48.2%), and 26.1% in the placebo group (follow-up 26.1%). The results are statistically significant (P<0.05); they show that in gonarthrosis pain the analgesic effect of verum acupuncture exceeds that of placebo acupuncture. Measurement of the functional parameters according to the Lysholm score showed no significant change. PMID- 18415454 TI - [Classification of patients with tension headache: irrationality, self communication, stress reactions and pain.]. AB - INTRODUCTION: A review of the literature suggests that patients with tension headache do not differ from normals as much as from each other in psychological traits. The aim of the present study was to identify psychological subtypes of tension headache patients. METHOD: The subjects were 130 inpatients with a diagnosis of recurrent tension headache who were treated in a pain clinic. Six factors measured the constructs irrationality, positive and negative self communication, emotionality, affective pain and situational pain. RESULTS: Hierarchical agglomerative cluster analysis allowed the identification of five subtypes of patients. The reclassification rate was 95%. All patients endorsed, to a high degree, pain adjectives that expressed obstinate-refractory pain qualities. Most, i.e. 81%, indicated that they suffered from affective pain: 51.5% reported high situation-dependent and affective pain. They were comprised by two clusters, 30% showing high irrationality/negative self-communication and low positive self-talk communication and 21.5%, the opposite profile. A further 30% of the patients suffered from affective pain but reported no stimulus dependence. They were divided into two groups: 17% reported high, whereas 13% reported low positive self-communication. In 18.5% of cases the patients reported no psychological causation and demonstrated a state of inner placidity and low irrationality. DISCUSSION: The results are discussed with reference to the literature and to indications for psychological treatment interventions. PMID- 18415455 TI - [Effects of intravenous clonidine on recovery and postanaesthetic analgesic requirements.]. AB - Pain and pain-related sympathoadrenergic reactions (hypertension, tachycardia) accompanied by nausea, vomiting and shivering are the most common side effects of recovery from anaesthesia. The alpha(2)agonist clonidine acts as a sedative, anxiolytic, antihypertensive, antiemetic, antisialogogue and decreases the incidence of shivering. Thus, we studied the effects of intraoperatively administered clonidine on the recovery period and the postoperative analgesic requirements in patients undergoing maxillofacial surgery. METHODS: After approval by the local Ethics Commitee and after informed consent had been given, 40 patients scheduled for elective maxillofacial surgery were included in this double-blind, randomized study. As a supplement to standardized general anaesthesia (isoflurane, N(2)O), the patients received either clonidine 5 mug/kg or placebo during the last hour of the operation. Blood pressure, heart rate, time of recovery, and sedation and pain scores were measured postoperatively. The occurrence of nausea, vomiting or shivering was noted, as were the requirements of piritramide for analgesia, which was administered on demand in titrating dosages, and of nifedipine for systolic blood pressure exceeding 180 mm Hg. RESULTS: The two groups were comparable regarding biometric parameters, ASA classification and duration of anaesthesia. Clonidinetreated patients were later in opening their eyes (22.5+/-11.9 min vs 17.9+/-10.9; n.s.) and the ability to state their dates of birth returned later (32.2+/-11.6 min vs. 25.7+/ 12.8;P<0.05). Pain was more frequent in the placebo group (P<0.05 after 30 min), and there-fore, these patients required much more piritramid (P<0.01). The sedation scores showed no significant differences. No vomiting occurred in the clonidine group, and shivering was less frequent (P<0.01). The placebo group received more nifedipine (P<0.05) because the rate-pressure product was higher (P<0.01). DISCUSSION: Opiates are frequently used as analgesics after maxillofacial surgery, even though their most common side effect-respiratory depression, nausea and vomiting-are particularly dangerous in these patients because of the obstruction of the upper respiratory tract. Self-titration of the opiate dosage on demand can decrease the incidence of serious side effects. Clonidine administered intraoperatively caused a profound reduction in analgesic requirements in this study. Additional opiate administration in the postoperative period was unnecessary in nearly all clonidine-treated patients. The attenuating effect on sympathoadrenergic reactions leads to lowering of the rate-pressure product and may be of advantage for patients suffering from arterial hypertension, angina pectoris or bronchial asthma. The slower emergence from anaesthesia following clonidine administration is probably caused by double-blind study properties preventing full consideration of the decreased isoflurane requirements after clonidine. PMID- 18415456 TI - [Postgraduate education in psychological pain treatment by the German section of the International Association for the Study of Pain.]. AB - Quality assurance with regard to pain treatment is a major concern of the German section of the International Association for the Study of Pain (IASP). For this reason, efforts to reach a consensus on guidelines for diagnostic and therapeutic procedures have been intensified and include educational endeavours as a basis for state-of-the-art practice. The German IASP section has implemented a postgraduate educational programme on pain treatment for psychologists. Objectives are to lay the foundation for a scientifically based assessment and treatment of chronic pain and to enable communication and cooperation among the disciplines. Admission is restricted to psychologists who have already started a course of training in psychotherapy, thus ensuring that sufficient knowledge about behaviour changes will be obtained in addition to the special programme about pain. After completion of the training in psychotherapy, a certificate will be issued if the student fulfils the following requirements: (1) attendance at 150 classes on pain syndromes, their anatomical and physiological basis, and their medical treatment, psychological methods of pain assessment and treatment, and effective ingredients of psychological treatment for pain; (2) experience in the treatment of patients with chronic pain; and (3) submission of 10 reports on assessments and treatments of chronic pain patients under supervision. To ensure training on a regular basis, the educational programme is offered to members of the IASP jointly by the German IASP and the Academy of the German Psychological Association. A sequence of eight modules covering the curriculum is repeated at least every 2 years, thus permitting entry to the programme at any time. PMID- 18415452 TI - [Pain therapy in herpes zoster and post-zoster neuralgia.]. AB - Herpes zoster neuralgia and post-zoster neuralgia (PZN) are common disabling pain syndromes. While pain from acute herpes zoster is self-limited in most cases, as pain may disappear without treatment, post-zoster neuralgia is difficult to manage. Pathological findings in acute herpes zoster include infiltration of ganglia, demyelinization and loss of axons; yet the pathogenesis of pain remains largely unknown. In postzoster (often incorrectly called post-herpetic) neuralgia, peripheral and central origins are mentioned for the development of pain: selective loss of myelin-sheathed nerve fibres, sensitization of peripheral nociceptors, cross-talk between afferents and sympathetic efferents, deafferentation with somatotopic remodeling, virus-induced spontaneous activity, and nociceptive nervi nervorum. Pain shows no sex-specific differences, but there is a clear predominance in elderly patients over 60 years of age. In these patients aggressive therapy should be instituted. Numerous pharmacological, anesthetic and surgical approaches have been proposed for the treatment of pain in herpes zoster. Most approaches have been studied in uncontrolled settings. Treatment is most effective when installed early in the course of the disease. For acute zoster pain, treatment with acyclovir, glucocorticosteroids and sympathetic blocks reveal the best results. PZN of less than 3 months' duration should be treated with sympathetic blocks. Long-standing PZN resolves in two thirds of the cases when treated with tricyclic antidepressants. TENS may be tried concomitantly. Topical ASA and capsaicin show promising effects and should be the object of further investigation. The same is true for specific zoster hyperimmunoglobulins and non-specific immunoglobulins; however, there are no definite results. In the future, controlled, double-blind studies on the effect of therapeutic measures in preventing postzosteric neuralgia need to be conducted. So far, the positive effect of sympathetic blocks in preventing the late pain complications of herpes zoster can only be suggested and recommended based on subjective experience. PMID- 18415457 TI - [Not Available]. PMID- 18415459 TI - [WHO step II-clinical reality or a didactic instrument?]. AB - INTRODUCTION: In 1986 the World Health Organisation (WHO) proposed an analgesic ladder for the effective therapy of cancer pain. The three standard analgesics making up this ladder are aspirin (non-opioid), codeine (weak opioid) and morphine (strong opioid). Adjuvant drugs may be added at any level. However, before 1986 step II analgesics (weak opioids) had never been tested in cancer pain relief. METHODS: This report presents a computer-assisted Medline (US National Library of Medicine) literature search restricted to the years 1986 1994, which was conducted to test the validity of the WHO guidelines, and in particular that of step II. RESULTS: We found seven retrospective studies and one prospective study on cancer pain treatment according to the proposed WHO guidelines that had been published since 1986. Every publication decribed the use of all three steps of the analgesic ladder. We found no prospective controlled trials demonstrating the efficacy and safety of WHO step II in particular. DISCUSSION: The use of the WHO guidelines "by mouth, by the clock and by the ladder" is now the mainstay of cancer pain management. Because of the guidelines' simplicity they found general acceptance and helped to establish an international pain therapy standard for worldwide use. Nevertheless, there is no scientific validation of WHO step II. In the absence of prospective controlled randomized trials additional longterm results are necessary. We need more data on the use of WHO step II and an update of the published guidelines taking account of modern sustained-release drugs. Up to now, step II of the WHO guidelines for cancer pain is not a clinical reality but at best a didactic instrument. PMID- 18415458 TI - [Drug therapy for tumor pain I. Properties of non-opioids and opioids.]. AB - Analgesic pharmacotherapy represents one of the major approaches to the treatment of cancer pain, since it is used in almost every patient. A thorough evaluation of the physical and mental status of the patient and of the pain is as necessary as a sound understanding of the pharmacokinetic and pharmacodynamic characteristics of the analgesics selected. The World Health Organization (WHO) has issued a basic 3 stage progression for the treatment of cancer pain, the "WHO Analgesic Ladder". Assignment to the stages depends mainly on the intensity of the pain rather than on its specific aetiology. Mild to moderate pain is treated with non-opioid drugs; moderate to severe pain, with a combination of a "weak" opioid and a non-opioid; and "strong" opioids should be used in combination with a non-opioid in the case of severe pain. Adjuvant drugs can be added if specifically indicated. Nonopioid analgesics include non-acidic compounds, e. g. paracetamol and metamizole, and acidic non-opioids, e. g. acetylsalicylic acid and newer non-steroidal anti-inflammatory drugs (NSAID). In contrast to most of the opioid analgesics, they have a ceiling effect for analgesia. Addiction and tolerance are extremely rare concerns. Opioids can be subgrouped into "weak" (e. g., codeine, dextropropoxyphene) and "strong" opioids (e. g., morphine) and also into drugs interacting with different opioid-receptor subtypes. Whereas pure agonists (e. g., morphine) produce increasingly intense analgesia with increasing dose, partial agonists and agonist-antagonists have a ceiling effect for analgesia and therefore have only a minor role in the treatment of chronic pain in cancer patients. Adverse effects occur in most patients in a dose-dependent manner. The most common of these is constipation; nausea, vomiting and sedation occur mostly at the start and can usually be treated effectively. The appropriate dosage, route of administration and dosage scheme of analgesics needs to be worked out for each individual patient in intensive work with the patient and a close follow-up, for years if necessary. Some analgesics may not be available in some countries, or only in specific preparations. PMID- 18415460 TI - [Experience with retrogasserian glycerol rhizotomy in the treatment of trigeminal neuralgia.]. AB - From August 1981 to May 1993 a total of 1263 percutaneous retrogasserian glycerol rhizotomies after Hakanson were performed. The intervention was performed with X ray monitoring under local anaesthesia and rarely lasted longer than 20 min. It achieved good results in the treatment of idiopathic trigeminal neuralgia (TN) and symptomatic trigeminal neuralgia due to multiple sclerosis (TNMS). Some 97% of the TN patients were completely free of pain after the intervention. The recurrence rate within 5 years was 12.8%. Some 94.7% of the TNMS patients were immediately free of pain, but within 2-5 years they experienced a high recurrence rate of 40.2%. The results in the patients with atypical facial pain were more disappointing: only 66.6% were pain-free immediately after the intervention, and the recurrence rate was 31%. With respect to the side effects immediately postoperatively, herpetic eruptions were found in 43.2% of cases. They occurred on the 3rd postoperative day and persisted for 2-3 weeks before being relieved by local virostatic therapy. Hypaesthesia and hypalgesia were present in the early postoperative follow-up in half of our patients. Both these reductions of sensitivity have a tendency to regress. Later, after 2 years, there was reduction in sensitivity of this type in only 20% of cases. In the follow-up 17.5% of our patients complained of dysaesthesia and in 21.4% corneal sensitivity was reduced or lost. We believe that glycerol rhizotomy, owing to its effectiveness, easy applicability, slight distress for the patients and low side effects, should be recommended as a first measure for non-conservative treatment of idiopathic trigeminal neuralgia as well as trigeminal neuralgia in multiple sclerosis. PMID- 18415461 TI - [Results of microsurgical decompression in trigeminal neuralgia.]. AB - INTRODUCTION: From 1 August 1983 to 6 June 1992, 284 patients underwent decompression of the trigeminal root in the rear part of the skull as treatment for tic douloureux. According to preoperative diagnosis and intraoperative inspection, a space-occupying process was the cause of the typical neuralgia in 13 cases (4 meningiomas, 3 epidermoid tumours, 3 acoustic neuromas and 2 trigeminal neuromas). In 271 cases (95.4%) microsurgical vascular decompression according to Jannetta was carried out. METHODS: The majority of patients were between 45 and 75 years of age. The follow-up period ranged from 10 months to 9.3 years (average 59.45 months). The results of the long-term investigations are based on standardized questionnaires completed by 202 patients. Of the total of 271 patients operated upon by Jannetta's technique, 261 (96.3%) were free of pain immediately after the operation. The main causes of failure were misdiagnosis (myoarthropathy, cluster headache) and incorrect indications (encephalomyelitis disseminata, trigeminal neuropathy following peripheral lesion). In 3 cases vascular displacement and complete decompression of the trigeminal root were impossible because the basilaris was exceptionally long. RESULTS: In the long term, 87.6% of the patients operated on remained free of pain or improved sufficiently to require no further carbamazepine medication. Relapses developed in 7.4% of cases, and except for operative revision and rhizolysis of the trigeminal root in a case of relapsed trigeminal neuralgia, thermo-controlled high-frequency lesion of the gasserian ganglion was carried out in a second operation. The complications of the Jannetta operation were hypoacusis and anacusis (4%), hyposmia (1%), dizziness (3.5%) and chronic subdural haematoma (1 case). Neither postoperative bacterial meningitis nor any lethality was recorded. In 94.5% of cases the patients expressed positive opinions of the operation and its results, while 4% regretted having the operation and 1.5% were undecided. The question as to whether the operation had significantly improved the quality of life was answered in the affirmative by 88.4% of the patients. DISCUSSION: In summary, the long-term results confirm that microsurgical vascular decompression can be offered as the method of choice for treatment of trigeminal neuralgia in younger patients, and in older patients when cardiopulmonary risk factors and cerebrovascular processes can be eliminated. Alternative methods are high frequency lesionsing of the gasserian ganglion according to Sweet and chemorhizolysis of the gasserian ganglion, but these must be restricted exclusively to the treatment of typical trigeminal neuralgia with tic douloureux. Persistent neuropathic pain caused by atraumatic or drug-induced lesion to the trigeminal nerve cannot be positively influenced either by surgical decompression or by destructive operations on the gasserian ganglion. PMID- 18415462 TI - [Prescription of strong opioids by general practitioners in Germany.]. AB - INTRODUCTION: Undertreatment of cancer pain is a well-known and worldwide problem. Every country has its specific restrictions. In Germany studies have referred undertreatment to pain states and therapeutic aspects in transverse inquiries, and others to 6-month records of regional prescription data. This study adds some long-term (3 years) information about doctors' (330 general practitioners throughout Germany) prescribing habits concerning strong opioids. METHODS: This analysis is based on data from 330 computerized practices whose prescription data are all turned over to the MIDOC (Medizinische Informations- und Dokumentations-Gesellschaft mbH, Weinheim). The data were evaluated for the period May 1990 to April 1993. RESULTS: Frequency of strong opioid prescription: The special prescription form for strong opioids ("Betaubungsmittelrezept" according to the narcotic drug law) was used in 0.026% of all prescriptions (Table 1). Of all cancer patients, 2.49% received at least one prescription for a strong opioid. Though the underlying pain states of the patients are not given among these data, there is no reason to suppose that German cancer patients will deviate to epidemiologically known pain severity and frequency (around 50% with severe opioid-sensitive pain, at least in the terminal phase). Prescription frequency in general practice: During the 3-year observation period, 37% of the practitioners used no opioid prescription forms at all, 38.6% used them for a single patient only, and in half of these, just once. The number of cancer patients per practice of opioid non-prescribers (81) did not differ from the average (85), but there was a wide range (0-592) in the number of cancer patients treated per practice during the 3 years. Dosage and administration interval: The most frequently prescribed strong opioid was the oral slow-release morphine tablet (45%), and the most frequent dose the 30-mg tablet; in more than 10% of cases the preparations prescribed were to be taken as required. CONCLUSION: We urgently need more information on the underlying reasons for the undertreatment of cancer pain. The data presented so far make it possible to draw up guidelines and a graduated education programme for patients and health care professionals. PMID- 18415463 TI - [Combined intravenous administration of diclofenac and apazone for postoperative analgesia A randomized study of 112 patients with access to i. v. on-demand analgesia after minor orthopaedic operations.]. AB - In a double-blind, randomized, placebo-controlled study, 112 patients scheduled for knee-joint arthrotomies or minor orthopaedic operations received 75 mg diclofenac, 600 mg apazone, the combination of 75 mg diclofenac and 600 mg apazone, or placebo (50 ml NaCl 0.9%) as a single i.v. dose immediately after operation. Postoperative pain intensity was measured by a numeric rating scale. All patients were allowed to self-administer piritramide from a PCA (patient controlled analgesia) pump (Prominjekt, Pharmacia, Sweden) in 2-mg boluses every 5 min during the first 6 h and subsequently every 15 minfor another 18 h after surgery. The patients receiving diclofenac, apazone, or the combination of diclofenac and apazone required a significantly lower cumulated dose of piritramide during the first 24 h after operation than did placebo-treated subjects (38 mg vs 39 mg vs 27 mg vs 67 mg;P<0.05), but there were no significant differences among the former three groups of patients. The incidence of typical side effects of opioids and antipyretic anti-inflammatory analgesics (nausea, vomiting, stomach ache, headache, vertigo) was low, and they were easily controlled in all cases. Postoperative combined application of the nonsteroidal anti-inflammatory analgesics diclofenac and apazone results in a significantly lower opioid requirement (about 60%) after minor orthopaedic surgery. The opioid sparing effect appears to be superior to that of diclofenac (44%) or apazone (42%) alone, but this was not statistically significant. PMID- 18415464 TI - [Severe respiratory depression caused by incorrect connection of a PCA pump. : A case report.]. AB - CASE REPORT: We report a case of severe respiratory depression during postoperative patient-controlled analgesia (PCA) in a 14-year-old boy. The medication cassette of a Pharmacia CADD-PCA 5200 was not properly connected, which led to a free-flow infusion of about 85 mg piritramide (strong mu-opioid agonist) within 15 min; the patient lost consciousness and developed apnea. He was successfully treated with artificial ventilation via ambu-bag and 0.2 mg naloxone i. v. The incident occurred approx. 2 h after the start of postoperative medication, when other infusions (suspended above the PCA device level) had been stopped, making the free-flow opioid infusion possible. As the PCA device was in a bedside pump enclosure, the disconnection was not immediately apparent. DISCUSSION: Although PCA is considered a safe method, it can have potentially lethal complications: Technical problems or serious handling errors involve the risk of large volumes of analgesics being infused within a very short time. Therefore, we recommend apparative monitoring (e. g., pulse oximetry) as a necessary condition for the safe use of PCA. PMID- 18415465 TI - [Low back pain/sciatica due to an osteoid osteoma.]. AB - A 21-year-old man suffered from diffuse low back pain and sciatica for 10-s periods once or twice a day over a period of 6 months. After this, pain became chronic and was resistant to conventional conservative treatment. Only acetylsalicylic acid diminished pain. Skeletal scintigraphy demonstrated a hot spot at the right side of the fifth lumbar vertebra, and CT scans showed a typical osteoid osteoma in the right pedicle of that vertebra. The time between onset of symptoms and final diagnosis was 18 months. Symptoms disappeared after surgery. Clinical and radiological aspects of the case are discussed. PMID- 18415466 TI - [Self-injection of local anaesthetics in chronic pain-a case report.]. AB - This is the first case report of a patient self-administering local anaesthetics to painful peripheral nerves in the face. Severe chronic pain of neuropathic origin was caused by a trauma of the face and head following a traffic accident 20 years ago. The patient himself chose the method of self-injection for pain control. He applies the local anaesthetic bupivacaine with an insulin syringe while looking in a mirror. PMID- 18415467 TI - [Not Available]. PMID- 18415469 TI - [Not Available]. PMID- 18415468 TI - [Not Available]. PMID- 18415471 TI - [The treatment concept at the "Center for Pain Studies" of the Rehabilitation Institute of Chicago.]. AB - The article describes the interdisciplinary treatment of chronic pain at the Center of Pain Studies (CPS). The CPS is an intergral part of the Rehabilitation Institute in Chicago. An essential part of the program is a 1-day outpatient evaluation in which members of all disciplines take part. A team conference determines the appropriateness of the pain program for each patient accepted for further care at the CPS. In 50% of the patients chronic pain is connected with work-related accidents. General aims of the treatment are to return to productive life, improve emplopyability, increase activity level, improve mobility, apply pain management techniques, and improve coping. The program is monitored by an evaluation/follow-up system. Success criteria of the 6-month follow-up for the years 1987-91, such as less daily downtime in 58-78% of the patients, more time out on weekends in 40-68%, return to work in 50-67% and reduced depression in 37 70%, give an impression of the effectiveness of the pain management program. PMID- 18415470 TI - [Clinical features, pathophysiology and treatment of fibromyalgia.]. AB - In rheumatology, all of the more than 400 specified syndromes are associated with pain. In the conceptual discussion on the multidimensional influences postulated to explain the development of chronic pain, fibromyalgia has gained increasing interest. Fibromyalgia (fibrositis) is an unspecific soft-tissue disorder with chronic wide-spread musculoskeletal pain and palpable hypersensitivity at fibrositic tender points. Fibromyalgia is often associated with fatigue, nonrestorative sleep and other symptoms. The syndrome has a high prevalence in women, but in most cases it has a long course with unsatisfactory attempts at therapy before the diagnosis is made. Though diagnostic criteria have been defined to describe it as a distinct clinical syndrome, speculations on its aetiology and pathogenetic mechanisms are still controversial. Histochemical investigations on muscle biopsy and biochemical tests have revealed unspecific changes but no characteristic muscle abnormality. It is supposed that the clinical features may result from central neurohumoral dysfunction combining with peripheral mechanisms to result in hyperalgesia. An integrated therapeutic concept with a reassuring and positive doctor-patient relationship can be helpful in achieving satisfactory treatment results. PMID- 18415473 TI - [Long-term efficacy and side effects of various migraine prophylactics: a retrospective analysis.]. AB - INTRODUCTION: Drug therapy for the prevention of migraine attacks is becoming more and attacks is becoming more and more important. The aims of such prophylactic treatment are to reach a lower frequency, shorter duration and milder intensity of migraine attacks, and to reduce the intake of anti-migraine medication, to improve the quality of life and working ability. The question of efficacy and tolerance of established migraine prophylactics [1] has been thoroughly investigated in many studies. So far the question of sustained efficacy after a successful prophylactic treatment completion has not been a research priority, but it is nonetheless of great importance. Researchers at the neurologic scientific research institute of the university of Naples have followed up migraine out-patients after successful prophylactic treatment and observed that prophylactic agents differ not only in their immediate efficacy and safety, but also in long-term efficacy. Therefore, an open pilot study was performed with the prophylactic agents propranolol, flunarizine, pizotifen, DHE retard, methysergide and cyclandelate in the recommended dossages (Tabe 1). OBJECTIVE AND METHODS: The aim of this study was to determine whether the various prophylactic agents available differ in active and long-term efficacy (at the end of a period after a successful prophylaxis=follow-up) and in the distribution of long-term responders at the end of the follow-up. The side effects of all prophylactic agents during active prophylaxis were also compared. Initially, 387 outpatients who had successfully completed a period of prophylactic treatment were recruited, and 208 were included in the study. At the time of follow-up a further period of prophylactic treatment was recommended if the efficacy rate was lower than 40% of the baseline at the end of the active prophylaxis period. The patients kept migraine headache daries (MHD) during the active prophylaxis and the follow-up, recording the following migraine objectives: number of attacks, pain total index (PTI), frequency of awakening with headache, and use of analgesics. RESULTS: The results showed that cyclandelate-actually a drug that is not yet officially accepted-had especially good results from the aspects of immediate efficacy, long-term efficacy and tolerance, compared with all other prophylactic agents. Significant differences were found in the duration of active prophylaxis. The mean monthly duration for patients treated with pizotifen (4.2), cyclandelate (3.9), and DHE retard (3.8) was longer than for those treated with flunarizine (2.8), and for patients treated with pizotifen it was longer than for those receiving propranolol (3.4). The mean duration (in months) of the postprophylactic period was distinctly longer for patients treated with cyclandelate (18.2) than for patients treated with DHE retard (12.9), flunarizine (13.1), propranolol (13.3) or pizotifen (13.8), but comparable with that after methysergide (17.2). Among the 208 patients, 85 were long-term responders (with no indication for repeated prophylaxis). No significant differences were found between the various groups, but the group of patients treated with cyclandelate was the only one with more than 50% long-term responders (18 vs 14). In general, the side effects of pizotifen, flunarizine and DHE retard seemed to be most pronounced. For cyclandelate, propranolol and methysergide fewer side effects were reported. CONCLUSION: In spite of the uncontrolled pilot design, it can be said in summary that all prophylactic drugs were effective. Cyclandelate had a good safety profile, and in efficacy it was at least comparable to the other prophylactic drugs. Patients treated with cyclandelate had a longer duration of active treatment and likewise a longer period of follow up. In addition, the proportion of patients with "no indication for repeated prophylaxis" at follow up was higher than for any of the other drugs. The results are interesting for medical practice and suggest replication in a randomized blind study. If the results yielded by the present study are confirmed, cyclandelate should be classified as a drug of first choice for migraine prophylaxis. PMID- 18415472 TI - [Prevalence of migraine in Germany.]. AB - INTRODUCTION: There are no valid data available for Germany on the prevalence of migraine using the new diagnostic and classification criteria of the International Headache Society (IHS). SAMPLE AND METHODS: Therefore, a survey on migraine prevalence was carried out on a representative sample of 2000 residents aged 16 to 69 years of the states of the former West Germany. The questionnaire incorporated the new criteria of the IHS and the diagnosis of migraine was based on the corresponding classification. RESULTS: Some 23.4% of the sample suffered from headaches, and 3.6% were identified as having migraine according to the IHS classification. The prevalence of migraine was 5.3% in women and 1.7% in men. The highest prevalence of 5.7% was found in the group aged 40-49 years. By extending the 4-72 h IHS attack duration criterion to 2-72 h, the migraine prevalence increased from 3.6% to 4.4%. DISCUSSION: For methodological reasons our study may underestimate the true prevalence of migraine in Germany. Taking our rather "conservative" figures into account, our study reveals, however, that there are more than 2 million migraine sufferers in Germany between 16 and 69 years of age. PMID- 18415474 TI - [Wound perfusion with bupivacaine for postoperative pain relief after elective abdominal surgery.]. AB - Local wound perfusion with bupivacaine after elective abdominal surgery seems to be a promising method of reducing postoperative pain. Applicability, effectiveness, side effects and risks were tested in a pilot study with 28 patients. During closure of the laparotomy incision two catheters were inserted, one between peritoneum and fascia and the other subcutaneously. After connection to bacteria filters bupivacaine was administered as boluses four times daily for 3 postoperative days, the dosage being adjusted to the patient's weight. Statistically significant pain reduction measured on the VAS was seen from day 1 to day 3 (immediately postoperative 4.92, day12.12, day 20.94, day 3 0.80). Additional analgesic medication was needed in three patients on day 1, after which pain treatment by catheter perfusion alone was sufficient. No severe side effects occurred, but the study was interrupted nonetheless in 3 patients, 2 of whom had a transit syndrome while the third had more severe hypotension than could be explained by the bupivacaine blood level. No infections and no problems with wound healing occurred during the study. This study proved that the method tested leads to significant pain reduction after elective abdominal surgery and has no severe side effects. PMID- 18415475 TI - [Long-term outcome of monoradicular lumbosacral compression syndromes-a comparison of pain and neurological deficits with lumbar CT scan.]. AB - A total of 97 patients with an acute monoradicular lumbosacral compression syndrome and a herniated disc at the same level were reinvestigated an average of 25 months after the acute event. Assessment was based on CT investigation and "root score", which was compiled from reports of subjective complaints and neurological deficits. All patients had initially undergone conservative treatment as inpatients. At the time of reinvestigation CT examination of the herniated discs revealed that they were unchanged in 56.7%, some-what smaller in 34%, no longer visible in 4.1% and larger in 5.2%. Nevertheless, 53.6% of patients were free of complaints, while 23.7% had improved clinically by more then 75%. The remainder showed improvement by about 25-75%. In no patient was the score unchanged or worse. In 69% of the patients neurological deficits were no longer demonstrable, and the remaining patients were not aware of deficits or did not feel any impairment from deficits that were obvious on clinical example. In 22.7% impairment was caused solely by local radicular or pseudoradicular pain. From these results it can be concluded that herniation of the disc precipitates the acute event but is not the sole source of the pain. In the majority of cases, during the further course of the disease the herniated disc causes no pain. It appears, therefore, that the course of disease following a disc herniation can be influenced very favourably by non-operative treatment and that in more than 50% of cases the condition becomes clinically silent. It follows that in neuroradicular compression syndromes a neurological deficit associated with the acute event does not mean operative intervention is indicated. PMID- 18415477 TI - [Not Available]. PMID- 18415478 TI - [Not Available]. PMID- 18415476 TI - [Brain tumor and headache.]. AB - METHODS: The possible association of brain tumour with headache was investigated in 100 patients seen for brain surgery. Preoperatively, 43 patients suffered from headache. These patients were thoroughly questioned about the nature of their pain. Investigation included the McGill Pain Questionnaire. RESULTS: In only 11 of the patients was headache the primary symptom of a brain tumour. Pain intensity was found to be lower in patients with brain tumour then in those with extracranial tumours or headache of other origins. Female subjects, patients under 50 years of age and those with elevated intracranial pressure experienced more intensive pain. Diurnal variation in pain intensity was observed in 60% of patients with headache. There was no evidence, however, of an association with elevated intracranial pressure. CONCLUSIONS: Our investigations yielded new information concerning the epidemology of headache accompanying brain tumours. Headache is not an early cardinal symptom of brain tumours, as was generally believed earlier. With the help of the McGill Pain Questionnaire a fine quantitative and qualitative characterization of headache of different origins could be made. The connection between tumour localization and pain lateralization, as well as the possible mechanisms of intracranial pain projection was extensively analysed. The interpretations of the results are at best hypotheses and they do not help determine why more than half of the patients with brain tumour did not experience headache. PMID- 18415479 TI - [Not Available]. PMID- 18415480 TI - [Not Available]. PMID- 18415481 TI - [Not Available]. PMID- 18415482 TI - [Not Available]. PMID- 18415483 TI - [Non-steroidal anti-inflammatory drugs (NSAIDs): Antirheumatic, anti-pyretic or analgesic drugs?]. AB - This manuscript reviews some aspects of the unorthodox development of analgesics. Most of them were not discovered by systematic research for analgesics; rather they were developed primarily as antipyretic or antirheumatic drugs that later became analgesics due to clinical observations. One reason for this development may be that the definition and quantification of analgesic effects under clinical circumstances is rather difficult and a clear dose-response relationship almost impossible to obtain. These drawbacks limit the discovery of new analgesics. On the other hand, modern insights into the site of action and the mode of action of antipyretic analgesics are likely to further the discovery of new and better analgesics. PMID- 18415484 TI - [Not Available]. PMID- 18415486 TI - [Not Available]. PMID- 18415485 TI - [PRN analgesic drug administration and PCA in children and adults following surgery for funnel chest.]. AB - Postoperative pain therapy often appears to be inadequate in children. The present study was a retrospective investigation of the administration of analgesics given to 133 children and adults following surgery for funnel chest between 1983 and 1994. METHODS: Pain therapy was performed with PRN (prore nata) administration in 100 patients (4-36 years) and with intravenous PCA in 33 patients (10-27 years). All narcotics and non-narcotics given up to the second postoperative day were evaluated. Total doses of narcotics were computed on a microgram per kilogram of body weight basis as morphine equivalents (ME) and total doses of non-narcotics, on a milligram per kilogram of body weight basis. RESULTS: In the PRN group significant differences in postoperative analgesia between children and adults were detected, but there were no differences between younger and older children. The number of narcotic administrations was lower in children (4-16 years) than in adults (7 vs 12.1 analgetic doses;P<0.0001). In adults significantly higher total doses of narcotics were administered than in children (100.5 vs 67.8 mug/kg body weight ME;P<0.05). Treatment with nonnarcotics was not different between children and adults. In the second part of the study 43 patients in the PRN group, who had been treated with narcotics exclusively, were compared with the PCA group. Children with PCA had a significantly higher total consumption of narcotics than children in the PRN group (146.8 vs 74.4 mug/kg body weight ME;P<0.001). No differences in narcotic doses were found in adults with PCA and those with PRN medication. There was wide individual variation in the use of narcotics in PCA patients, but no differences between children and adults. CONCLUSION: Children received smaller quantities of narcotics than adults when postoperative analgesia was performed with PRN administration. PCA led to delivery of larger quantities of narcotic drugs than PRN medication in children. PMID- 18415487 TI - [Pain treatment with dihydrocodeine slow release. Results of a post marketing surveillance study.]. AB - Chronic pain requires chronic treatment. Dihydrocodeine retard (DHC) complies with the requirements for treatment of chronic pain: its sustained release formula provides pain relief for up to 12 h. Thus, taking two tablets of this preparation daily is sufficient to ensure continuous pain relief. Patients and methods The 309 physicians participating in the study treated a total of 1502 patients and provided more than 5000 reports containing data on pain relief and side effects. Treatment was performed according to the WHO principles for the analgesic treatment of cancer pain patients, i.e., doctors and patients were taught that two tablets are taken per day, one in the morning and one in the evening, irrespective of whether pain was present or not. Most of the data were derived from the first 4 weeks of treatment. Patients were selected at random. All had chronic pain, but the diagnosis was not a selection criterion. The patients had had prior treatment with various analgesic regimens, and in most cases drugs were administered at irregular intervals, i.e., on demand. About half of the patients (54%) suffered from pain related to the musculoskeletal system such as back pain, joint pain, polyarthritis. Twenty-four percent had cancer pain and 22% had pain caused by other sources, mostly neuropathic pain, including cases of severe postherpetic neuralgia. Most of the patients were older than 50 years; the average age in the patient population was 62 years. There were 816 women and 686 men. Patients assessment of analgesic treatment was performed before starting therapy with DHC (thus conferring to prior therapy) and after 1 and 2 weeks of treatment with the new drug. While only about 10% of the patients found their prior pain treatment excellent or good, nearly 80% rated the treatment with DHC as excellent or good and only 2% as bad.Severity of pain was assessed by the patients on a four-step verbal rating scale ranging from "no or little pain" to "extremely strong pain". At the time of admission to this post marketing surveillance 51.5% of the patients suffered from very strong pain and 41% reported strong pain. Five percent had extremely strong pain and only 2.1% reported no or little pain. After 2 weeks of treatment with DHC, 54.5% had little or no pain, 29% suffered from strong pain, 7% from very strong, and 0.4% from extremely strong pain.Sleeping problems are known to be reported by patients with chronic pain. They often cannot sleep continuously for more than a few hours. Thus, the effect of DHC on sleep was evaluated. Before starting the new treatment only 8% of the patients were having uninterrupted sleep for 6 h or more, and more than 50% of the patients slept less than 3 h. During treatment with DHC 48% of the patients had more than 6 h of uninterrupted sleep and about 82% slept continuously for more than 3 h per night. Side effects About 20% of patients reported nausea at baseline (during previous treatment); vomiting was reported in 7.6%. These percentages did not change during the first week of treatment with sustained-release DHC and even decreased slowly during the next 3 weeks. The frequency of constipation increased from 14% at baseline to about 29.5% at the end of the second week of treatment with DHC with no change during the next four weeks. A total of 312 side effects were mentioned in 5308 reports delivered by the 1502 patients during the treatment with DHC (including multiple reports). The most frequent side effects were gastrointestinal (n=106), followed by symptoms related to the central nervous system such as dizziness, sedation, etc. (n=50), and non-specific symptoms such as indisposition (n=29). Other specific symptoms were rare and distributed over many different organ systems. Insummary, the findings of this post marketing surveillance study suggest that sustained-release dihydrocodeine is an effective and safe analgesic drug for the treatment of chronic pain of various causes. PMID- 18415488 TI - [The clinical picture of the chronic pain patient-Analysis of a pain outpatient clinic using headache as an example.]. AB - INTRODUCTION: Patients with chronic headache are seen in an interdisciplinary pain clinic after many years of treatment. Compared with other pain syndromes, the standards for diagnosis and treatment are widely accepted according to the guidelines of the International Headache Society. Nevertheless, many patients continue to suffer. Analysis of their special clinical features may help to clarify what kind of conditions potentially cause chronicity. PATIENTS AND METHODS: A total of 193 patients were seen in 1992-1993 in our pain clinic. Epidemiological, medical and psychological data were analyzed retrospectively. Classification of headache syndromes and pain behavior was based on the criteria of the International Headache Society, taxonomy of the Multiaxial Pain Classification System for Somatic Dimensions (MASK-S) and "Stages of Chronicity". Important psychodynamic factors were taken from standardized psychological interviews. RESULTS: The patients had a mean age of 46 years; 67 % of them were female. Fifty-three percent of the patients suffered from chronic tension headache; a high percentage was suspected to be caused by analgesic intake. Because of their pain behavior, 68 % were classified as being in an advanced chronic stage with a poor prognosis. The most important findings of the psychological diagnosis was that 40 % of patients had a depressive style of conflict solving. CONCLUSIONS: The results are discussed with respect to their relevance in explaining development of chronic pain, as seen in selected headache patients in an interdisciplinary pain clinic. PMID- 18415489 TI - [Quality assurance in therapy of chronic pain. Results obtained by a taskforce of the German Section of the Association for the Study of Pain on psychological assessment of chronic pain. V. Instruments for the assessment of pain-related cognitions and coping with pain.]. AB - The present paper is one of a series of publications, reviewing German instruments for psychological assessment of pain. Their main focus is on the results of a task force on quality testing for each subject. This paper describes and comments on methods regarding self-reporting of pain cognitions and both cognitive and behavioral strategies for coping with pain. Concerning pain cognitions one focus is on patients' attributions of causes of pain and the modes of controlling pain (subjective pain model). The other focus is on instruments recording "pain beliefs" in the sense of dysfunctional congitions associated with the experience of pain. Each instrument was examined with reference to approved psychometric criteria, empirical foundation and clinical relevance. It was noted that several instruments are deficient in their psychometric criteria and their empirical foundations. We used these data as a basis to elaborate a specific and differential recommendation. A similar procedure was followed with instruments for the assessment of pain-related coping strategies. According to our research there are two subgroups of coping instruments, one more specifically for cognitive coping with pain, and the other combined with behavioral coping strategies. Once again, we elaborated a specific and differential recommendation, giving priority to instruments taking account of both cognitive and behavioral dimensions of coping with pain. PMID- 18415490 TI - [Not Available]. PMID- 18415491 TI - [Not Available]. PMID- 18415492 TI - [Not Available]. PMID- 18415493 TI - [Not Available]. PMID- 18415495 TI - [Calcitonin gene-related peptide immunoreactive nerve fibres in the dura mater encephali of the rat: experiments related to the neurogenic inflammation of meningeal structures.]. AB - In this paper morphological and physiological experiments are described that refer to the concept of neurogenic inflammation of meningeal structures as a putative source of migrainous pain and other headaches. The main emphasis of this study carried out on the duramater encephali of the rat was the functional role of calcitonin generelated peptide (CGRP), a vasodilatory neuropeptide of fine afferent nerve fibres. Immunocytochemical preparations showed that the parietal dura mater was densely innervated by CGRP immunoreactive nerve fibres, the distribution and ultrastructure of which were examined by ligh and electron microscopy. The dense innervation around the medial meningeal artery suggested a vasomotor function of these peptidergic fibres. In further experiments the CGRP immune product of the nerve fibres could be diminished by electrical stimulation of the dura mater. Extracellular recordings from trigeminal ganglion cells showed that electrical and mechanical stimulation of large dural vessels activate trigeminal afferents. In a final series of experiments the dural blood flow around branches of the medial meningeal artery was measured with a laser Doppler flowmeter. The blood flow was increased by electrical stimulation of the dura, the size of this effect depending on stimulus strength and frequency. This increase was inhibited in a dose-dependent manner by the competitive CGRP antagonist CGRP(8-37), which shows an involvement of CGRP in the regulation of meningeal blood flow. We conclude that stimulation of trigeminal afferents innervating the dura mater releases CGRP from peptidergic afferent terminals, thereby causing vasodilatation and increasing the meningeal blood flow, an important component of neurogenic inflammation. The preparation decribed will be used for further studies on basic mechanisms of neurogenic inflammation and nociception in meningeal structures. PMID- 18415494 TI - [Pharmacotherapy of cancer pain : 2. Use of opioids.]. AB - The adequate use of opioids in the treatment of chronic cancer pain requires sound knowledge of selection criteria for the various opioids, the routes of administration, dosages, dosing schemes and possible side effects. Drug selection depends on the intensity of pain rather than on the specific pathophysiology. Mild to moderate pain can often be treated effectively by so-called "weak" opioids. These include codeine, dihydrocodeine and dextropropoxyphene. Non-opioid analgesics, like acetylsalicylic acid or paracetamol can be added according to the "analgesic ladder" proposed by the World Health Organization (WHO). If adequate pain relief is not achieved "strong" opioids are required. The route of administration that is the safest and the least invasive for the patient should be chosen. Non-invasive (oral, rectal, sublingual, transdermal and intranasal) and invasive routes (intravenous, subcutaneous, spinal and epidural) are available (Table 8). Noninvasive routes are preferred, and most patients can be maintained on oral opioids. Alternatively, in some patients pain can be managed by the sublingual (buprenorphine) route. A transdermal preparation exists for fentanyl, but has not yet been approved for the German market. If the oral route cannot be used or if large doses are required, it will be necessary to change to an invasive route. Intravenous bolus injections provide the fastest onset of analgesic action. They are mostly used in very severe pain. Repeated injections can be avoided by using intravenous or subcutaneous infusions. Various types of pumps delivering analgesics at constant basal infusion rates with the option of rescue doses in case of breakthrough pain are available (patient-controlled analgesia=PCA). Opioids frequently used for s. c. infusion are morphine and hydromorphone. Adjuvant drugs (antiemetics, anxiolytics) can be added. Epidural or intrathecal administration of opioids should only be used in intractable pain or if severe side effects, such as sedation and confusion, will arise with systemic opioids. Morphine, hydromorphone, fentanyl and sufentanil have been used, as have other additional compounds (e.g. local anaesthetics, clonidine). Intracerebroventricular application of morphine has been used only occasionally. In all cases, opioids should be given on to a fixed time schedule thereby, preventing pain from recurring. Additional rescue doses (approximately 50% of baseline single dose) are given for break-through pain. The most frequent side effect of opioids is constipation, and the administration of laxatives is often recommended (Table 5). Nausea, vomiting, sedation and confusion mostly occur in the beginning of opioid therapy. In contrast to constipation, tolerance to these effects develops within days or weeks. True dependence or psychological addiction rarely occurs in patients with chronic cancer pain. In most cases, progression of the underlying disease associated with increasing tissue damage and increasing pain is found. Fear of dependence and addiction often contributes to undertreatment of patients suffering from chronic cancer pain. PMID- 18415497 TI - [The vertebrogenic factor and thoracalgia.]. AB - INTRODUCTION: Reversible functional joint disorders (joint blockages, somatic dysfunction) of the intervertebral and rib joints can be treated using manual therapy, which improves the related segmental changes in both the dorsal and the ventral area [pseudoradicular syndrome, hyperalgesia zone (HAZ)]. This phenomenon is triggered by a decrease in the heightened nociceptor irritation in the joint capsule and in the surrounding tissues. METHODS: The patients were examined lying prone to assess the functional rib and intervertebral joint disorders. The deep, periarticular tender points directly surrounding intervertebral joints displaying functional disorders were infiltrated with 5 ml lidocaine 0.5% in four segments in 35 patients. In a further 48 patients the same local anaesthetic dose was used, but given by intracutaneous infiltration in the area of painful thoracic segments. A third group of 31 patients was treated manually in accordance with current practice. RESULTS: Ventral HAZ and also pain disappeared in 26 of the patients (74%) who received deep lidocaine infiltration. Comparable results were achieved with manual therapy (in 68%). However, only 8 of 48 patients (17%) became painfree after intracutaneous lidocaine infiltration (p<0.05) Freeing of the rib joint blockages was observed in 15 of 25 patients after deep lidocaine infiltration, in 9 of 31 patients after manual therapy, and in 3 of 8 patients after intracutaneous lidocaine infiltration. DISCUSSION: From our results we conclude that blocking of the peripheral nociceptors, rather than of the peripheral nerve bundles, is of primary significance for the effective treatment of anterior thoracalgia. PMID- 18415496 TI - [Craniomandibular disorders and the cervical spine syndrome as differential diagnoses in suspected idiopathic trigeminal neuralgia.]. AB - INTRODUCTION: Unfortunately, sharp, severe pain in the area of distribution of the fifth cranial nerve is frequently termed trigeminal neuralgia, and no differentiation is made between typical and atypical neuralgia and other types of facial pain disorders. This can lead to inadequate treatment. CLINICAL MATERIAL AND METHODS: From 1987 to 1993, 577 patients were referred to our clinic with the diagnosis "idiopathic trigeminal neuralgia". Re-examination of these patients was based on a comprehensive history, behavioural and psychosocial assessment, general inspection of the head and neck, evaluation of the craniomandibular system and cervical spine, neurovascular, neurosensory and motor evaluation of cranial nerves, intraoral evaluation, head and cervical spine scans (CT and/or MRI), diagnostic anaesthetic injections, laboratory tests and/or response to therapy. RESULTS: The follow-up examinations and treatment results allowed confirmation of the diagnosis in only 55% of all cases. There were 82 patients (14%) with craniomandibular disorders, 54 patients (9%) with cervical spine syndrome and 18 patients (3%) with cervical and/or dental causality in addition to the trigeminal neuralgia. CONCLUSION: The process of differential diagnosis is critical in trigeminal neuralgia, because an incorrent or missed diagnosis is one of the most frequent causes of treatment failure. As idiopathic trigeminal neuralgia, craniomandibular disorders or the cervical spine syndrome can involve similar symptoms and response to the use of medication, close interdisciplinary cooperation in the process of diagnosis is recommended. PMID- 18415498 TI - [Quality assurance in therapy of chronic pain. Results obtained by a taskforce of the German Section of the Association for the Study of Pain on psychological assessment of chronic pain.]. AB - This is the first in a series of publications presenting the results of a taskforce on quality assurance in psychological assessment of chronic pain. The initiative was motivated by the increasing and confusing variety of newly developed German instruments and/or translations of Anglo-American instruments. Our main work was therefore concentrated on the collection of existing German assessment instruments, on summarizing the essentials in a documentary sheet, and on examining their objectivity, reliability, validity, clinical relevance, economy and degree of empirical foundation. For each diagnostic domain we thus elaborated specific differential recommendations for those working in psychological pain research and clinical practice, in an attempt to devise criteria enabling them to choose the optimal instrument or test battery for their needs and conditions. PMID- 18415499 TI - [Trigeminal neuralgia-how patients experience their condition disease.]. AB - INTRODUCTION: Effective treatment of patients with trigeminal neuralgia is often a long and complicated procedure. The symptoms of trigeminal neuralgia are clearly defined in most cases. Sudden and brief episodes of severe and stabbing pain (tic douloureux) occur, with pain usually starting from a trigger point. Recent reports suggest 80-90% suppression of pain with various treatment regimens, which seems to indicate that the diagnosis and successful treatment of the disorder are no longer a major problem. In fact, however, the intense suffering of patients and isolated reports in the literature suggest that there are still considerable diagnostic difficulties. Patients are referred from one specialist to another, in most cases without the necessary interdisciplinary cooperation, and countless interventions and attempts at therapy not only remain unsuccessful, but may cause serious adverse effects. METHODS: The records of 120 trigeminal neuralgia patients in two different areas of Germany were analysed retrospectively. In addition, the course of the disorder from the initial symptoms up to the institution of effective therapy was documented by means of information obtained from the general practitioner or other physicians. The patients themselves were also interviewed about their history. RESULTS: After a so-called pain career of 51/2 years, each patient had taken an average of 4,000 pills or capsules, 2,500-3,000 of which were carbamazine, lost 1-2 healthy teeth in attempts to treat the painful condition, undergone three or four maxillofacial or ENT operations, or been subjected to exeresis. The result is prolongation and exacerbation of suffering and an apparently hopeless situation. The financial burden on the patients and the health insurance companies is enormous. DISCUSSION: Apparently there is a considerable need for more information about the clinical symptoms, cause, diagnosis and therapy of trigeminal neuralgia, especially as the symptoms are often no longer typical because they have become chronic or are the result of previous treatment. This is needed by all specialists involved, including dentists and general practitioners. In patients in whom clinical criteria suggest the diagnosis of trigeminal neuralgia, drug treatment should be initiated immediately in consultation with the neurologist or neurosurgeon. For cases in which drug treatment fails or resistance to the drug develops surgical treatments are available, such as non-destructive microvascular decompression or thermocoagulation of the gasserian ganglion. PMID- 18415501 TI - [Not Available]. PMID- 18415500 TI - [Not Available]. PMID- 18415502 TI - [Not Available]. PMID- 18415503 TI - [Not Available]. PMID- 18415504 TI - [Not Available]. PMID- 18415505 TI - [Not Available]. PMID- 18415506 TI - [Not Available]. PMID- 18415508 TI - [Not Available]. PMID- 18415507 TI - [Not Available]. PMID- 18415509 TI - [Not Available]. PMID- 18415511 TI - [Not Available]. PMID- 18415510 TI - [Not Available]. PMID- 18415513 TI - [Not Available]. PMID- 18415512 TI - [Not Available]. PMID- 18415514 TI - [Not Available]. PMID- 18415516 TI - [Not Available]. PMID- 18415515 TI - [Not Available]. PMID- 18415517 TI - [Not Available]. PMID- 18415518 TI - [Combination analgesics consisting of paracetamol plus acetylsalicylic acid: benefits and risks.]. AB - According to the German Drug Law of 1976 the present status of scientific knowledge regarding the benefit/risk-ratio of combination analgesics was reevaluated and is summarized taking the fixed combination of paracetamol plus acetylsalicylic acid as an example. The extensive discussion of the responsible committee for reevaluation of drugs B-3 (Neurology/Psychiatry) at the German Federal Institute of Health (Bundesgesundheitsamt) led to the following results as viewed by the involved members: - the fixed combination has its pharmacological rationale (secure detoxification, even with high single doses; (over)additive analgesic effect in experimental models. - the benefit of the fixed combination is given by a potentially lower liver toxicity as proven with experimental models and by an (over)additive efficacy in cancer pain or headache. - combination-specific risks surpassing the ones of the single substances are not recognizable. This holds particularly true also for the risk of analgesic nephropathy being presented elsewhere [19]. Even though the clinical efficacy has been proven only in few specific studies the evaluation of the benefit/risk-ratio appears to be positive regarding the not recognizable combination-specific risks. The combination is recommended for acute use. PMID- 18415520 TI - [Systemic administration of alpha(2)-adrenoceptor agonists for postoperative pain reliefagonists for postoperative pain relief.]. AB - The alpha(2)-adrenoceptor agonist clonidine has analgesic properties comparable to those of opioids after systemic administration. It also has antihypertensive, antiemetic, anxiolytic, sedative and antisialogogue effects and reduces the incidence of shivering. Thus, the pharmacodynamic profile of clonidine seems to suit it quite well for the special problems related to recovery from anaesthesia. The respiratory depression is weak, particularly in comparison with opioids. This paper reviews previous experience with systemic administration of alpha(2) adrenoceptor agonists for postoperative pain relief. Especially in combination with low-dose opioids, clonidine leads to a similar or even better level of pain relief with significantly reduced adverse side effects compared with opioid mono therapy, perhaps because different sites of action are addressed and influence nociception. Therefore, balanced postoperative analgesia including alpha(2) adrenoceptor agonists, therefore seems to be a beneficial addition to differentiated postoperative pain relief. PMID- 18415519 TI - [Antinociceptive effects of alpha(2)-adrenoceptor agonists ("analgesic" actions in animal experiments)agonists ("analgesic" actions in animal experiments).]. AB - alpha(2)-Adrenoceptor agonists like clonidine, dexmedetomidine, and ST-91, inhibit nociceptive reflex activity predominantly by a spinal mode of action. They mimic the action of the inhibitory transmitter noradrenaline, which is released from the terminals of bulbospinal monoaminergic pathways. The inhibition by noradrenaline is due partly to hyperpolarization of the postsynaptic neuronal membrane; however, the selective antinociceptive effect of the alpha(2) adrenoceptor agonists results from reduction of the release of the excitatory transmitters such as glutamate and substance P, blockade of the binding of substance P to spinal neurones, and enhancement of the action of the inhibitory transmitter, 5-hydroxytryptamine. Clonidine and dexmedetomidine stimulate adrenoceptors of the alpha(2A) subtype, while ST-91 stimulates alpha(2B) adrenoceptors. Antinociception is manifested not only by depression of nociceptive reflexes and behaviour, but also by inhibition of the expression of immediate early genes in dorsal horn neurones following noxious stimulation. The inhibitory control from the brain stem of spinal nociceptive activity can be triggered by alpha(2)-adrenoceptor agonists. Moreover, impulse conduction in C fibres of peripheral nerves is far more reduced by these compounds than that in A fibres. Antinociceptive effects are reported to occur in various models of clinical pain, e.g. the formalin test, adjuvans-induced arthritis, autotomy following deafferentation, and "hyperalgesia" after nerve ligation. Therefore, the mechanisms involved in antinociception may also be responsible for the analgesia produced by alpha(2)-adrenoceptor agonists. PMID- 18415521 TI - [Quality assurance in therapy of chronic pain. Results obtained by a taskforce of the German Section of the Association for the Study of Pain on psychological assessment of chronic pain. VIII. Instruments for the assessment of pain associated dimensions: depression, somatic symptoms, anxiety.]. AB - Chronic pain is often associated with an array of general psychological dimensions such as depression, somatic symptoms, anxiety, anger and a loss of quality of life. Part VIII describes various instruments concerning each dimension and comments on their applicability within a diagnostic routine for chronic pain patients. TheAllgemeine Depressionsskala (German version of CES-D) is recommended for the assessment of depression, and theBeschwerden-Liste seems best for assessing somatic symptoms. Trait-anxiety and pain-unrelated state anxiety seem not to be suitable because they lack significance for the therapy process. While the inventory for assessing anger still needs an empirical basis for chronic pain patients, quality of life inventories are not recommended for routine pain assessment. PMID- 18415522 TI - [High-dose intrathecal clonidine in the treatment of neuropathic tumor pain. Two case reports.]. AB - Two cases with perineal pain caused by recurrent carcinoma of the rectum are reported. Initially both patients suffered from predominantly nociceptive pain, which was treated adequately with spinal opioids. Tumor growth with epidural spread and infiltration of the plexus lumbosacralis caused severe neuropathic pain. Both patients were free of pain with a combination of spinal clonidine and opioids. Clonidine doses had to be increased up to 1.31 and 1.46 mg daily in order to provide adequate analgesia. Outpatient treatment was possible for several weeks with stable dosage. Bradycardia and hypotension occurred with initial dose titration and after dose increases and were treated with parasympathicolytic drugs and vasopressor agents. Both patients were given spinal clonidine until their death 4 1/2 and 4 months later. In the final stages, adjuvant systemic administration of morphine was necessary to control dyspnea. PMID- 18415523 TI - [Not Available]. PMID- 18415524 TI - [Not Available]. PMID- 18415525 TI - [Not Available]. PMID- 18415526 TI - [Not Available]. PMID- 18415527 TI - [Not Available]. PMID- 18415528 TI - [Dangers and complications in pain therapy with epidural and intrathecal catheters.]. AB - Pain therapy with epidural or intrathecal catheters is an invasive method. These techniques have specific indications in both acute and chronic pain therapy. However, complications can occur. Thus, the potential complications and the therapy necessary must be known.Drugs: Complications resulting from acute local anesthetic intoxication's are rare. High plasma levels during chronic therapy may lead to confusion. Respiratory depression can occur in opioid naive patients up to 12 (-24) h after injection. Adequate monitoring is a prerequisite for this therapy. After application of clonidine, hypotension is frequent in hypertonic and hypovolemic patients. Epidural or intrathecalcatheter placement can result in therapeutic failure, trauma by punction and inability to place the catheter. During chronic therapy, technical problems can occur, e.g., dislocation, occlusion. To exclude intrathecal and intravascular placement, application of a test dosage of a local anesthetic with adrenaline is recommended.Neurological complications can result in nerve root deficit or "simple" post-spinal headache, but cauda equina syndromes, paralyses, intracranial bleeding, sinus thrombosis and central neurological deficits have been reported. Skininfection at the insertion site of the catheter has been observed with an incidence of 1.9 to 7.7%. A spinal infection with neurological deficit is rare. Spinal infections are often associated with other diseases. Spinalhematomas are rare. Coagulation disorders and anticoagulants can lead to bleeding. Intravenous heparin should be avoided, because this is frequently associated with spinal bleeding. Therapy with cumarines is a contraindication for insertion of spinal catheters.Monitoring: During treatment with spinal catheters, adequate monitoring increases safety for the patients. Efficacy of the injections, puncture site and the neurological status should be documented daily. Neurological deficits must be diagnosed without losing time and adequate therapy must be initiated. PMID- 18415529 TI - [Diagnosis and treatment of atypical facial pain-a review.]. AB - Atypical facial pain is a collective term used for otherwise unclassifiable pain syndromes of the face. It describes mostly unilateral lancinating and burning pain, which is constantly present but fluctuating in intensity and may be accompanied by dys-or paraesthesias over a period of several years. Women are more often affected than men. A strikingly large number of invasive investigative procedures are performed before the final diagnosis is established. Psychopathological abnormalities are frequent and do not consist solely in masked depression. To date there is no cure for atypical facial pain. Most frequently tricyclic antidepressants such as amitriptyline and imipramine, MAO inhibitors or anticonvulsives such as carbamazepine and phenytoin are used, which only alleviate the pain. This also holds true for behaviour therapy, which should be the first-line treatment. Any invasive intervention should be avoided. In the differential diagnosis atypical facial pain should be distinguished from cervicogenic or cluster headache, and also from intracranial tumours of the trigeminal nerve or the cerebellar-pontine angle, erosive tumors of the base of the skull, orbitas and nasopharynx. PMID- 18415530 TI - [Quality assurance in therapy of chronic pain. Results obtained by a taskforce of the German Section of the Association for the Study of Pain on psychological assessment of chronic pain : VI. Instruments for the assessment of disability VII. Psychological instruments for the assessment of pain relevant aspects of social interaction and communication.]. AB - Disability is a central aspect in the assessment of chronic pain patients. Disability questionnaires in German (developed or adapted) are examined and selected for different purposes. The "Funktionsfragebogen Hannover" and the "Pain Disability Index" are recommended for both research and clinicalapplication while the "Sickness Impact Profile" is suitable only for research purposes.Four different German questionnaires for the assessment of social interaction and communication in pain patients and their significant others are examined in part VII. There are some empirical data for three of them. Only the "Inventory of Familial Adaptability and Cohesion" has achieved a certain degree of empirical maturity. Further research and developmental activity in this area of pain assessment are urgently needed. PMID- 18415531 TI - [Chronic, sympathetically maintained pain syndrome after mild lesion of the brachial plexus-a case report.]. AB - The present article is a case report on a chronic sympathetically maintained pain syndrome following a mild lesion of the brachial plexus. Apart from slight motor and sensory disturbances, which disappeared within a few months, no autonomic, motor or sensory deficit remained. Spontaneous pain and a remarkable allodynia in the region of the left upper quadrant, always without the left hand developed. The pain syndrome obviously extended beyond the region of the primary affected nerve. In the climax of the pain attacks it became manifest like a mirror reflexion on the opposite side. The diagnostic and therapeutic problems that occurred during these years and the dependency of the pain attacks on exterior influences are evaluated. PMID- 18415532 TI - [Spinal epidural abscess-an interdisciplinary emergency.]. AB - Introduction Spinal epidural abscess is a rare cause of neurological disability, which is frequently, preceded by local spinal pain. There are only a few case series in the literature and the prognosis for this disease has not improved significantly over the past 50 years. Patients and methods Seven patients were treated neurosurgically with laminectomy and abscess drainage for spinal epidural abscesses from 1991 to 1994. Results All but one patient had several weeks of local spinal pain (mainly after minor trauma), immunosuppression, and all exhibited excessively high blood sedimentation rates. On admission they presented either with hemiparesis or with signs of complete spinal cord transection. Since preoperative neuronal damage persisted too long, only 3 patients showed slight improvement of their neurological functions postoperatively. Conclusion A spinal epidural abscess is a rare, but devastating interdisciplinary emergency, which poses no operative technical problem to the neurosurgeon. Poor results can be avoided by carefully taken individual histories, which are dominated by long standing and significant local spinal pain in most cases. The pre-operative neurological status is crucial for the long-term outcome. Elevated blood sedimentation rates in conjunction with local pain of the vertebral column should direct attention to the possibility of an epidural abscess. Magnetic resonance imaging is the radiological technique of choice for establishment of the diagnosis. Conventional X-rays are of no value for early diagnosis because neurological signs and symptoms precede bony changes of the vertebrae. PMID- 18415533 TI - [Not Available]. PMID- 18415534 TI - [Not Available]. PMID- 18415536 TI - [Two-years' follow-up after lumbar disc surgery.]. AB - INTRODUCTION: Outcome studies of lumbar disc surgery document a success rate ranging between 49% and 90%, according to the evaluation criteria used. The aim of the present study was to investigate the prognostic value of pain history and of sociodemographic, psychodiagnostic and medical factors for the long-term outcome after lumbar disc surgery. METHODS: Between selection for and performance of operations for herniated lumbar disc, 100 patients took part in standardized interviews about their clinical and social history. During the same session they answered one questionnaire on depression and one on coping strategies in response to pain. Two years after the operation, 83 patients answered the standardized follow-up questionnaires. RESULTS: During the follow-up period, 7 (8.4%) patients had further back surgery, and 20% of the patients reported severe back pain on the follow up. With multivariate discriminant analysis, correct prediction of at most moderate back pain or severe back pain 2 years after the operation had been possible in 94.3% of the patients. The important predictors were found to be: duration of subjectively reduced working capability, pain at other locations, consideration or application for disability pension, depression, and severe acute pain. Risk factors for further back surgery were pain at other locations, guest for social support in response to pain, absence of muscle palsy and of Lasegue's sign. CONCLUSION: Anamnestic information on the pain history and psychological factors have prognostic value for the outcome after lumbar disc surgery. If there is no absolute medical indication for disc surgery, we suggest reconsidering the treatment planned for patients with the risk factors specified. PMID- 18415535 TI - [Pharmacotherapy of cancer pain. 3. Adjuvant drugs.]. AB - Adjuvant analgesics are drugs that are not primarily used as analgesics but can produce analgesia in certain types of pain. Adjuvant analgesics can be administered together with non-opioid and opioid analgesics on each step of the WHO analgesic ladder. They should be given when an additional or specific indication exists, but should not be used as a substitute for a thorough treatment with opioids and nonopioids. Adjuvant analgesics can be classified into groups according to the type of pain to be treated: continuous neuropathic pain or lancinating neuropathic pain, sympathetically maintained pain, bone pain and those for multipurpose use. Adjuvant drugs used for continuous neuropathic pain include local anaesthetics, clonidine, capsaicin, and antidepressants. Tricyclic antidepressants are the group that have been best investigated, and are therefore the drugs of choice. An analgesic effect is probably produced via enhancement of transmitter concentrations in pain-modulating pathways. This occurs at lower doses than those necessary to treat depression. Anticholinergic actions, acute glaucoma, constipation, orthostatic hypotension and cardiac arrhythmias are adverse effects that are seen predominantly with teritiary amine drugs and less often with secondary amine compounds. Initial doses should be small to avoid these adverse effects. Local anaesthetics are used less often, because of the high incidence of side effects (especially with tocainide, flecainide). An analgesic effect has been described in neuropathic pain, however, probably due to membrane stabilization and reduction of aberrant signal conduction. Mexiletine is considered to be the safest local anaesthetic, and should be used initially in small doses (100-150 mg/d). If side effects do not occur, doses can be increased step-wise up to 900 mg/d. Local anaesthetics are indicated for the treatment of severe neuropathic pain; this treatment is contraindicated in patients with cardiac arrhythmias. Systemic or intrathecal clonidine can be tried in neuropathic pain refractory to opioid therapy. The same stands for the topical application of capsaicin in certain types of pain. Lancinating neuropathic pain is an indication for anticonvulsant drugs. Carbamazepine, clonazepam, valproate and phenytoin seem to reduce aberrant signal conduction in damaged nerves in a manner similar to the supression of epileptiform activities in the brain. Common side effects include sedation, dizziness and nausea. Of greater concern are the more severe side effects, such as bone marrow depression (carbamazepine) and hepatotoxicity (phenytoin, valproate). Low initial doses and stepwise increases in dosage, repeated blood counts, and monitoring of plasma levels are helpful in recognizing and avoiding these adverse effects. Baclofen, a GABA agonist primarily used for spasticity, is effective in the treatment of trigeminal neuralgia and is often used in the management of lancinating pain of unspecific origin. The initial dosage is 10-15 mg/d, increasing to 30-90 mg/d, or higher. If neural blockade fails to reduce sympathetically maintained pain sufficiently specific adjuvants can be used. Sympatholytic drugs, e.g. phenoxybenzamine (60 120 mg/d) or prazosin, can be administered to patients without major cardiovascular dysfunction. There is experimental evidence of the involvement of calcium channels in nociception, and a beneficial clinical effect of nifidepine in reflex sympathetic dystrophy (RDS) has been demonstrated. Bone pain is common in tumor patients and can often be treated effectively with non-steroidal anti inflammatory drugs. Biphosphonates (etidronate, clodronate, pamidronate derivates) also produce analgesic effects in patients with bone metastases. However, differences among the various compounds have not been clearly evaluated yet. Potent and specific radioisotopes are still under development and the use of calcitonin in bone pain is considered controversial. PMID- 18415537 TI - [Experience with intra- and perioperative high-dosage steroids in microneursurgical revision operations on lumbar discs.]. AB - A consecutive series of 100 microneurosurgical revision operations on lumbar discs has been evaluated in a prospective and randomized clinical trial analyzing the influence of dexamethasone on postoperative clinical outcome. METHODS: Of the 100 patients, 50 received dexamethasone 8 mg by local intrathecal injection during the operation and perioperative dexamethasone by a systemic route for 7 days. A reference group of 50 patients did not receive any steroids. The two groups were compared with reference to postoperative lumbar and radicular pain, regression of paresis, analgesic requirements during hospitalization and duration of stay in hospital. One year after the end of the study clinical results and ability to work in the two groups were compared. RESULTS: A significantly lower analgesic requirement was found in the dexamethasone-treated group. We found no evidence that steroids had ameliorate the regression of paresis or lumbar and radicular pain at the end of hospitalization. There was no difference in the clinical outcome 1 year after reoperation. CONCLUSIONS: After revision operations on lumbar discs, corticosteroids are a helpful adjuvant drug for relief of lumbar or radicular pain, but they do not significantly improve the final clinical outcome. PMID- 18415538 TI - [A device for patient-controlled intranasal analgesia (PCINA).]. AB - INTRODUCTION: It has been demonstrated that intranasal opioid titration has a rapid onset of action and can provide satisfactory management of postoperative pain [10, 12, 14]. In these studies the intranasal titration was carried out by the investigator. Self-administration of an opioid intranasally by patients requires a spray bottle with safety precautions of an equivalent standard to those offered by an intravenous PCA device. We describe a device for patient controlled intranasal analgesia (PCINA) that meets these safety requirements. METHODS: The Baxter PCA on demand system consists of a mechanically driven infusor, a flow restrictor, and a patient control module for bolus administration. The flow restrictor provides a flow rate of 5 ml/h or 2 ml/h. This Baxter intravenous PCA system has been subjected to a slight modification to adapt it for PCINA. The patient control module has a bolus volume of 0.5 ml and in this modification it is attached, instead of to an intravenous line, to a narrow, 26-gauge plastic cannula with the needle tip removed (Fig. 1). To check the accuracy of the volume delivered, three PCINA devices with a flow rate of 5 ml/h (filling time of 6 min for the 0.5-ml bolus volume) and three PCINA devices with a flow rate of 2 ml/h (filling time of 15 min for the 0.5-ml bolus volume) were examined at defined time intervals. The PCINA devices were filled with distilled water and the volume demanded was immediately determined by means of a high-precision scale. Three determinations of the volumes delivered were performed. In an initial unblinded pilot observation in five orthopaedic patients, PCINA (for a 4-h period) was compared with the conventionally prescribed pain medication (for a subsequent 5-h period). For intranasal opioid administration, fentanyl (1 ml=0.05 mg) was used. At every evaluation point, pain intensity was evaluated with the aid of a 101-point numerical rating scale (0 = no pain, 100 = worst pain possible). At the end of both examination periods (PCINA/conventionally prescribed pain medication), overall patient satisfaction with the method of pain management experienced was evaluated (graded: very good, good, satisfactory, bad, very bad, not acceptable). RESULTS: The volumes delivered from the three PCINA devices with a flow rate of 5 ml/h (PCINA device 6') and from the three PCINA devices with a flow rate of 2 ml/h (PCINA device 15') are presented in Fig. 4. The subjective pain intensities measured with the 101-point numerical rating scale are demonstrated in Fig. 5. The patients used 0.28+/-0.097 mg fentanyl (0.15-0.45 mg) during the 4-h period of PCINA. No patients had any difficulty using the PCINA device. No technical problems arose with any of the devices. No patient complained of intranasal pain or burning during or after nasal administration. At the end of the study overall patient satisfaction with PCINA was judged as very good (2 patients), good 2 patients) or satisfactory (1 patient). The relief obtained with the customarily prescribed pain medication was judged as satisfactory (1 patient) or bad (4 patients). CONCLUSION: We conclude that the PCINA device presented fulfils the PCA device safety requirements. The bolus volume delivered by the device is precise and follows the manufacturer's specifications for flow rate and bolus volume. Initial. PMID- 18415539 TI - [Patient-controlled analgesia in outpatients with severe cancer pain.]. AB - In this case report, we describe continuous subcutaneous infusion of opiates as PCAO (patient controlled analgesia in outpatients) in one patient with metastatic carcinoma of the rectum (liver and bone metastases, partial bowel obstruction) with severe cancer pain and vomiting in the terminal phase. The parenteral administration of opioids extended over 58 days. The infusion was powered by an external portable clockwork-driven syringe pump (Perfusor M, Braun Medical/Germany). The open-accessible pump has a syringe volume of 10 ml, and its maximal infusion time is 24 h. The 27-G infusion needle (Sub-Q-Set, Baxter/USA) was inserted in the side of the abdomen and was left in the same position for 10 to 20 days. It took the patient and his family only 1.5 h to familiarize themselves with the use of the pump. They were trained in its use in our outpatient pain department. For pain control both the variable continuous infusion and the extra injection doses could be administered by the way of the syringe driver. The patient was given a stock of 120 ampoules of morphine for further treatment at home. For optimal pain control he decided to raise the daily dose of opioid infusion from the initial 60 mg to 240 mg morphine within 48 h. In this way, PCAO-besides rapid titration of the opioid dose to achieve analgesia allows the use of opioids controlled by the patient himself. In the present case this procedure was also important when an outpatient radiation therapy became urgently necessary to prevent a fracture of the spine because of metastasis. The pain control by the patient himself was the main factor to get free of pain during the transport to the hospital. Even positioning for radiation was possible without pain. When he received outpatient radiation therapy the patient needed extra injection doses of up to 360 mg morphine a day. The PCAO procedure by continuous subcutaneous infusion with opiates is a safe and efficient method of pain management for outpatient patients suffering from severe cancer pain and intractable nausea in the terminal phase. Its validity has also been proven especially for radiation treatment of bone metastases. PMID- 18415540 TI - [Interdisciplinary cooperation in prevention and treatment of low back pain.]. AB - An expert committee of the German Chapter of the IASP has published five recommendations for the prevention and early treatment of low back pain. These refer to an early activation of the patient facilitated by scheduled pain medication and an interdisciplinary treatment including physiotherapy and psychotherapy if risk factors for chronicity are detected. PMID- 18415541 TI - [Not Available]. PMID- 18415542 TI - [Not Available]. PMID- 18415543 TI - [Not Available]. PMID- 18415544 TI - [What can pain diaries achieve? Advantages and limitations of their use with reference to single case evaluation.]. AB - Pain diaries are modern evaluation instruments for assessment of pain dynamics and treatment outcome. There are syndrome-specific and non-syndrome-specific diaries. They exist as paper-and-pencil versions, as electronic diaries or as uptime recorders. Pain diaries are especially suitable for considering pain course over time. There-fore, they have many fields of application, which extend to the topography, differential diagnosis, therapeutic evaluation, classification, indications and, obviously, to questions of research. A particular advantage of pain diaries is the possibility of individual design and realization. Despite numerous advantages, problems with the application and data analysis of diaries must be kept in mind. They are instruments with a large potential reactivity of measurements, e.g. answer tendencies, memory effects, simulation. In addition, items in pain diaries usually have different scale levels, which can lead to problems in adaequate analyses of data. The data analysis of pain diaries is demonstrated with the help of selected individual cases. Three different phases are distinguished, and useful suggestions for analysis of individual cases are submitted. The first evaluation step includes the visual inspection of primary data. This provides a survey of the process of pain and at the same time a reference level for assessment of the secondary data. The secondary data can be divided into agglomeration, variation and correlation data. Agglomeration data, such as frequencies, sums or means, are a first quantitative, but still superficial, process of analysis. Additionally, the findings should be complemented by variation data to characterize these results with regard to their variability. The significance of deviation measures is demonstrated by a selected case example. A small deviation in this case reveals a relatively constant course of pain parameters, whereas large deviation reflects an unstable course. Moreover, the variability of data can be analysed by trend coefficient and time series analysis. Time-related measures of deviation, developed in the 1930s by von Neumann et al., and a simple version of time-series analyses developed by Tryon, are described and compared with one another. Finally, the use of correlation statistics is useful to reveal important relationships between secondary data. For the case example such statistically significant relations exist between pain intensity, therapeutic interventions, and sleep duration. Due to the effort required by diary analysis the use of pain diaries seems to be indicated only if there is the intention to generate quantitative data by mathematical-statistical methods. Problems arise, in spite of the quantitative data level, from the limited comparability of results and the undeniable reactivity of measurements. At present open questions refer to the desirable or non-desirable stability of surveys by means of pain diaries and the possible generation of secondary effects as the result of long-term diary recording. PMID- 18415545 TI - [Continuous subcutaneous buprenorphine application in the treatment of cancer pain.]. AB - Introduction Buprenorphine is well known in cancer pain therapy because of the long duration of its action and high analgesic potency. Many studies exist about the intravenous and sublingual application form; however, few data are available on its use by the continuous subcutaneous route. Methods Twenty-five patients were analysed retrospectively over 956 days who has been treated with continuous subcutaneous buprenorphine for cancer-related pain. In 7 of these 25 patients plasma analyses were performed. Due to a modified sensitive HPLC method with electrochemical detection for the analysis of buprenorphine in plasma, a detection limit of 40 pg/ml could be obtained. The other analytical methods for plasma concentration have detection limits between 150 and 500 pg/ml. Results During the treatment with continuous subcutaneous buprenorphine it was necessary to increase the initial average daily dose of 1.07 (+/-0.41) mg to 1.58 (+/-0.58) mg. The initially high pain intensity (rated from 0 to 100%) of 67% could be reduced to a moderate pain of 26% on average. Only 2 patients had to be switched over to morphine because of insufficient analgesia. In no case did complications occur that required intervention or would have made it necessary to change the pain therapy. Eighty percent of the patients judged this kind of treatment as effective and comfortable. Most often patients complained about drowsiness, low appetite and constipation. Because of the progress of the cancer disease these effects could not clearly be related to treatment side effects. With 7 of 25 patients the median daily dose of 1.2 (minimum 0.9-maximum 2.3) mg buprenorphine was related to the median plasma concentration of 438 (minimum 64-maximum 3374) pg/ml. In one case with progressive liver dysfunction, the potential risk of cumulation with buprenorphine could be controlled with this method. Conclusions Continuous subcutaneous buprenorphine with external infusors is a safe and efficient cancer pain therapy without severe side effects. Because of its ceiling effect, it is not as effective as morphine, but can be discussed as an alternative if other opioids cause incompatibility reactions. PMID- 18415546 TI - [Double blind study of intravenous aspirin vs placebo in the treatment of acute migraine attacks.]. AB - INTRODUCTION: The empirical use of oral acetylsalicylic acid (ASA) in the treatment of acute migraine attacks has led to the recommendation of ASA as a basic treatment of migraine. However, there are no controlled studies on the intravenous use of acetylsalicylic acid. METHODS: Forty patients with a 1- to 12 year history of migraine were included in this placebo-controlled trial. The sociodemographic data of patients receiving placebo or 500 mg ASA i.v. and migraine symptoms before treatment were identically distributed. Pain relief during attacks was documented by the Visual Analogue Scale (VAS) for 1 h at 5-min intervals. Furthermore, the change of concurrent non-pain symptoms was evaluated by self-rating (worse-unchanged-better-absent). RESULTS: The VAS showed a mean pain reduction of about 60% after 1 h versus placebo. Efficacy varied significantly between patients. We observed a small number of socalled non responders as well as patients with complete pain relief. The most common concomitant symptoms were nausea, photophobia, vertigo, vomiting and sweating. During the time of observation these symptoms were significantly reduced compared to placebo. The study shows a maximum placebo effect of about 17% (mean value). Significant differences in pain ratings were measured 25-30 min after application (P=0.0008). CONCLUSIONS: In this study i.v. ASA showed a significant reduction in pain intensity during migraine attacks. In addition, most concomitant non-pain symptoms were equally reduced. The onset of pain relief did not correspond with the inhibition of platelet aggregation (after 2-4 min) or with peak plasma concentrations of ASA (immediately after application) or salicylic acid (after 2 3 h). More efficacy studies with higher doses and longer periods of observation are necessary to optimize the treatment of migraine attacks with intravenous ASA. Studies including neurophysiological and vascular parameters are recommended to confirm the results of this study. PMID- 18415547 TI - [Development of a screening questionnaire to predict good and poor outcome of lumbar disc surgery.]. AB - AIMS OF THE STUDY: The aims were: (1) to investigate the prognostic value of pain history, sociodemographic, psychodiagnostic and medical factors for long-term outcome after lumbar disc surgery and (2) to develop a screening checklist of reliable predictors that distinguish between good and poor outcomes. METHODS: In six different spine centers 510 patients were assessed after indication for operation by independent research teams. Most of the patients (n=400) were being operated upon for the first time (group 1), while 110 patients had had previous back surgery (group 2). More than 80% of the patients participated in the follow ups performed 6 and 12 months later. RESULTS: During the follow-up period, 19 (4.8%) patients of group 1 and 11 (10%) patients of group 2 had renewed back surgery. The outcomes 12 months postoperatively were: good in 51.5%, moderate in 28.4% and poor in 20.1% for group 1; good in 43.2%, moderate in 32.1% and poor in 24.7% for group 2. In order to identify relevant predictors, the preoperatively obtained data of group 1 patients with a good outcome and those with a poor one were compared. Using multivariate discriminant analysis, 80% of these patients were correctly classified. The most important predictors were found to be: intensity of restricted physical mobility, number of other pain locations, duration of subjectively reduced working capability, and duration of acute back pain. On the basis of this statistical analysis, 11 predictors were selected and converted to scores, reflecting their relative significance. The calculation of a sum score gave an appropriate prediction of 76% for good and 79% for poor outcomes. For group 2 the results were nearly the same. IN CONCLUSION: It is possible to determine for each patient a predictor sum that is prognostic for the individual risk of a poor operation outcome. PMID- 18415548 TI - [Slow-release morphine liquid suspension for the therapy of cancer pain and non cancer pain-A pilot study.]. AB - INTRODUCTION: For pain therapy different methods of application are essential, because side effects or swallowing difficulties may prevent the natural oral application of analgesics. Up to now only tablets have been available for sustained-release morphine. We investigated a suspension of this drug in different pain states. METHOD: Slow-release morphine liquid suspension was administered for 3 weeks in two different groups of patients. Group I included patients previously on a combination of non-opioid analgesics and weak opioids with unsatisfactory pain relief (pain scores > 5 of VAS 0-10). Group II were patients with well-controlled pain on strong-acting opioids. In these patients, the medication was changed to slow-release morphine liquid suspension. Intensity of pain, capacity, subjective feelings and side effects were recorded throughout the investigation period. RESULTS: A total of 18 patients were included in the study, 8 in group I, 10 in group II. Twelve patients suffered from cancer pain. A pain reduction of at least 50% was achieved in 6 of 8 patients in group I. In 4 of 10 patients in group II the pain relief already obtained was improved further. In 8 of 18 patients the study had to be terminated. The reasons for this were unsatisfactory pain relief, increasing side effects distaste on intake. The average daily dose of morphine in group I was 132.5 mg, 151 mg in group II. Capacity did not change under slow-release morphine suspension. Constipation was the most, frequent side effect (67%). Ninety-three percent of all patients described the sweetish taste of the liquid preparation as unpalatable. DISCUSSION: These first results show that liquid slow-release morphine suspension can provide pain relief similar to slow-release morphine tablets. Consequently, slow-release liquid morphine suspension provides a suitable alternative, especially in patients with swallowing disorders. PMID- 18415549 TI - [Evaluation of third molar surgery by patients choosing between local anaesthesia alone and with additional conscious sedation.]. AB - This study was conducted on patients undergoing third molar surgery to evaluate their opinions on surgery and the follow-up period. Two groups were formed, as patients were able to choose between local anaesthesia alone or with additional conscious sedation by means of intravenous Midazolam. METHODS: A total of 426 patients ranging in age from 11 to 61 years (mean 20.8 years) participated, 335 of whom preferred Midazolam sedation (0.1 mg/kg) while 91 voted for local anaesthesia only. During the operation an impacted lower third molar was removed by osteotomy, and in 80% of these cases the upper third molar on the same side was also removed. A questionnaire on the operation, the follow-up period, postoperative pain and use of analgesics was distributed before the event. RESULTS: Women and younger patients preferred conscious sedation. Surgery was described as significantly less distressing by the sedated group (diagram 1), whereas there was no difference in views of the follow-up period or postoperative pain. Patients in the Midazolam group took more analgesics on postoperative days 1-3. Nonsedated men tended to evaluate surgery as more distressing than women in the same group, while there were no sex differences in the Midazolam group. CONCLUSION: Following the evaluation of surgery as more "pleasant" by sedated patients, it might be expected that this would contribute to a similar experience of the follow-up period. In this study, however no such connection was found. It is possible that preoperative self-selection of the patients (more sensitive and cautious persons preferred conscious sedation) might be responsible for these results. PMID- 18415550 TI - [Quality assurance in therapy of chronic pain. Results obtained by a taskforce of the German Section of the Association for the study of Pain psychological assessment of chronic pain. Instruments for the assessment of pain experience and pain behaviour.]. AB - The series of publications is continued in which the results of a review of German pain-related psychological assessment tools are presented. The taskforce of the German Society for the Study of Pain (DGSS) describes and examines instruments assessing the qualitative aspects of how pain is experienced (part II). Part III describes and comments on methods regarding self-reporting and observation of pain behaviour. Part IV discusses the use of pain diaries and introduces different forms of diaries, focussing on the measurement of pain intensity. PMID- 18415551 TI - [Not Available]. PMID- 18415552 TI - [Editorial.]. PMID- 18415553 TI - [Problems in and approaches to the design and evaluation of therapeutic studies in patients with headache.]. AB - A review of studies on migraine therapy shows a large heterogeneity in the clinical evaluation of different drugs and behavioral approaches. The percentages of efficacy of beta-blockers or behavior therapy (relaxation, biofeedback) range from 30% to 80%. Methodological differences from one author to another in the design and conduct of the studies might be causes of this variation. For both clinical practice and empirical research (e.g. replication of treatment studies) an uniformity in the application of different methodological techniques should be sought. This paper discusses the influence of methodological aspects on the success rates of treatment for chronic headaches, especially migraine. We question the present practice of applying traditional group experiments in headache research. Double-blind and placebo-controlled studies are often required, but this demand often cannot be niet in empirical reality because of the individuality of the patient or the heterogeneity of the symptoms. We discuss some alternative approaches that could be used in empirical research on headache therapy. The application of single-case experiments as well as methods of time series analysis are described as a more appropriate approach to the evaluation of studies on chronic headaches. PMID- 18415554 TI - [Therapy for migraine-the current state of the art and perspectives.]. AB - Migraine is a syndrome and not a nosological entity. It is therefore relatively improbable that a uniform etiology can be defined, and it must be assumed that there are different multifactorial etiological conditions for each individual. It is probable that a therapy concept that is equally valid and promising for all patients can therefore never be developed. On the one hand, there is a variable palette of therapeutic possibilities with a fixed differential indication with regard to the different variants of migraine, and on the other subtle analysis of individual cases is necessary by means of recording the individually relevant somatic, psychological, familial and social parameters. A treatment strategy must then be developed on an individual basis, in which both elements are taken into account. Therapy studies are carried out to investigate various questions and, accordingly, may have different experimental paradigms. The intrinsic action of a therapy method should be compared to the placebo effect on a randomized doubleblind basis. If this preconditions is fulfilled, appraisals of the real chances of success in the practice can be made more accurately on the basis of large-scale open studies comprising a representative cross-section of migraine patients. Further topics for therapy studies should include the analysis of responders and nonresponders and the development of differential indications for certain methods of therapy. PMID- 18415555 TI - [The "pain pen", a semiautomatic morphine injection system for subcutaneous self administration. Technique and first results.]. AB - Subcutaneous self-administration of morphine with a "pain pen", a semiautomatic mechanical injection system about the size of a penholder, was tested in nine patients with severe chronic pain of malignant origin. During a period of treatment of 12-100 days (on an outpatient basis in three patients) side-effects of subcutaneous morphine could only be observed during a short-lasting initial period of adaptation. The average daily dosage of morphine was between 11.3+/-7.2 mg (lowest daily demand) and 20.7+/-10.5 mg (highest daily demand). Pain scaling (NSR 101) demonstrated a significant pain reduction from 80+/-19.3 to values between 5.5+/-8.3 (lowest score) and 39.4+/-14.2 (highest score during subcutaneous therapy). There were no hygienic problems, and the patients learned the technique of self-injection within a few days. This system may be especially suitable for patients with gastrointestinal incompatibility of oral opiate preparations, e.g. patients with multiple drug intake. PMID- 18415556 TI - [Pain syndromes in tick-borne neuroborreliosis. Clinical aspects and differential diagnosis.]. AB - Tick-borne borreliosis (Borrelia burgdorferi) is a common and complex disorder affecting the skin, the joints and the nervous system. It progresses through different clinical stages. The clinical spectrum of neuroborreliosis has expanded since the introduction and widespread application of specific serological tests. We have investigated 41 patients with Bannwarth's meningopolyneuritis (MPN) as the classical form of neuroborreliosis, in a prospective (26 patients) and a retrospective (15 patients) study. When questioned, 19/41 patients reported a tick bite and only 15/41, erythema migrans as the characteristic early skin lesion. In 34/41 patients typical MPN characterized by painful radiculoneuritis and/or cranial neuritis, especially facial palsy, were seen. Among these, 3 had a complicated form with a progressive remitting relapsing course or focal central nervous system involvement (hemiparesis, cerebellar syndrome); 2 had mild meningitis and facial nerve palsy bilaterally without radicular pain; and in 5 radicular pain was the only symptom. MPN associated with Lyme arthritis was observed only once. In 2 patients in the retrospective study (no antibiotics in the acute stage) we saw a chronic spinal cord disorder with spastic paresis several years after uncomplicated MPN, accompanied in 1 of them by acrodermatitis chronica atrophicans (ACA), the typical late-onset borrelia-induced dermatosis. In the acute stage of the disease 40/41 patients had a cerebrospinal fluid (CSF) syndrome compatible with MPN (mononuclear pleocytosis, blood-brain barrier dysfunction, elevated IgG and/or oligoclonal bands). IgG antibody titers against borrelia antigen were elevated in all patients in the serum and in 21/30 also in the CSF. In all patients pain was an early and prominent symptom; the first symptoms are usually felt in the region of the tick bite or the erythema, initially as diffuse myalgia, arthralgia or pain in the connective tissue. In the further course the migrating pain becomes more radicular in character, without being limited to defined dermatomic areas or peripheral nerves. The intense, burning pain is characterized by exacerbation during the night. Peripherally and centrally acting analgesics have only minor effects. Often neurological deficits are still absent at this time. Erythema migrans with radicular pain in the region of the dermatological lesion was observed in 2 patients. This is an early manifestation of MPN. After MPN and/or Lyme arthritis a sympathetic reflex dystrophy (SRD) developed in 2 patients. In a further patient SRD was observed right at the beginning of the illness, immediately before MPN. There is a close clinical similarity between SRD and the acute stage of ACA. Therefore, borreliosis can be assumed to produce a painful skin dystrophy like SRD or ACA by direct injury to the sympathetic nerves even in the early clinical stage of the infection. The main conditions to be considered in the differential diagnosis are polymyalgia rheumatica; lumbar disk herniation; inflammatory radiculopathies of other origin (e.g. herpes zoster); painful neuropathies, including the diabetic thoraco-abdominal form; internal disorders of chest and abdomen with referred pain; lymphocytic meningitis of other origin, encephalomyelitis; and sympathetic reflex dystrophy. High-dose penicillin G i.v. is a potent analgesic in all patients with tick-borne neuroborreliosis. PMID- 18415557 TI - [Lateral asymmetry of the human pain sensitivity.]. AB - In repeated clinical studies a preponderance of pain syndromes on the left side of the body has frequently been observed. Experimental studies in humans revealed a lower pain threshold on the left, nondominant side. On the other hand, some studies do not confirm this lateralization. Since pain threshold is not a very valid measure of pain sensitivity in the range beyond threshold, and since clinical studies are limited by simply counting the incidence of the pain syndromes, we investigated whether a significant lateralization of pain sensitivity exists in the entire range beyond pain threshold. Handedness and gender were included as factors. For experimental pain stimulation in 24 subjects three different methods were used: local pressure on the middle phalanxes, a modified submaximal effort tourniquet test, and submerging the hands into cold water. For pain measurement beyond threshold we used the category splitting procedure (Heller).All three methods of stimulation produced corresponding results. Lefthanded subjects showed decreased pain sensitivity on the left side, right-handed subjects on the right side. This was true for the total range of pain sensitivity. For pain induced by pressure, lateral asymmetry increased with pain intensity, for the other two methods it was constant. Lateral asymmetry was found in all subjects, but significant differences could only be demonstrated in female Ss. It is concluded that both gender and handedness contribute to lateral asymmetry of pain sensitivity in man. PMID- 18415559 TI - [Not Available]. PMID- 18415558 TI - [Psychological aspects of acute postoperative pain.]. AB - Acute postoperative pain has an important psychological component. This psychological element could be registered by observing or measuring interindividual differences in pain experience, expressions of pain, and pain coping behavior. Emotions such as anxiety, anger, and helplessness accompany postoperative pain, and postoperative pain can also be elicited by very intense preoperative anxiety. This emotionally triggered pain exists independent of the sensory pain component elicited by the surgical injury and must be treated differently. Psychological factors also influence pain reduction. Postoperative aggression and its underlying metabolism facilitates recovery if the physicians or nurses can cope with the anger of the patient. Providing sensory descriptions of the pain during the preoperative visit and careful attention to both the pain experiencing and the pain-free patient help to reduce pain or prevent its escalation. PMID- 18415560 TI - [Not Available]. PMID- 18415562 TI - [Not Available]. PMID- 18415561 TI - [Not Available]. PMID- 18415563 TI - [Not Available]. PMID- 18415564 TI - [Not Available]. PMID- 18415565 TI - Editorial. PMID- 18415566 TI - [Documentation of chronic pain-a seroey.]. AB - This article reviews the methods currently in use for the measurement of chronic pain. The most important items for inclusion in questionnaires about the history and in pain diaries to elicit data on the time-course of pain are presented, and both the aims and the advantages and disadvantages of various strategies are discussed. The documentation of chronic pain in outpatients would allow answers to some questions concerned with medical epidemiology if practiced in a large number of therapeutic institutions, especially if the data were processed and evaluated by microcomputer. PMID- 18415568 TI - [Non-orthodox methods in pain therapy: Harmless and successful?]. AB - As a result of changing public attitudes non-orthodox methods of pain therapy have become widely accepted within everyday health care. The view that non orthodox methods, even if not overly successful are at least harmless is widely held. The present paper gives examples that support the existence of three risk factors: (a) Delay in the diagnosis of curable diseases owing to alternative, non orthodox forms of therapy. (b) Occurrence of serious or even life-threatening side-effects of non-orthodox forms of pain therapy. (c) Development of unusual diseases which are not easily diagnosed by the physician because he or she does not know that so-called natural or herbal products may be the reason for the symptoms. It is felt that these problems with non-orthodox pain therapy should be known by the physician. Patients should be questioned about the use of alternative methods of treatment and physicians should be aware of unwanted effects of acupuncture, cell therapy, Ajurvedic medicines and herbal therapies. In addition, they should know that some drugs sold as herbal products with "no side-effects" are enriched with e.g. phenylbutazone and/or corticoids, for example. PMID- 18415567 TI - [Postoperative pain.]. AB - A short survey about the different methods available for producing postoperative analgesia is given, the goal being to make it clear to the clinician that there are quite a number of techniques to be used although the everyday clinical practice often sticks to simple and not too effective methods of pain treatment following surgery. Initially presenting short informations about the neurophysiology of pain and the pathogenesis and causes of postoperative pain two main groups of producing analgesia are then discussed.Thefirst group deals with the systemic use of analgesics be it nonnarcotic analgesic antipyretics or narcotic analgesics (opioids). As for the first subgroup the peripheral action of these drugs (metamizol, acetylsalicylic acid, paracetamol) is brought about by blocking the synthesis of prostaglandins. These substances can only be used for very moderate postoperative pain f.i. following head and neck surgery. The strong acting opioids belong to the second subgroup. Recent informations on receptor sites in the brain and cord and the subgrouping of the receptors throws new light on the understanding of the different effects of these drugs and on the pathomechanisms of agonistic, antagonistic and mixed activities. The clinically used opioids then are mentioned (morphine, fentanyl, methadon, pethidin, piritramide, tilidin, buprenorphin and pentazocine) and dosage, duration of action, antagonisms and untoward side effects are presented. Stress is laid on the recent development of patient-controlled analgesia with all its advantages. Thesecond main group of methods for postoperative analgesia consists of regional anesthesia techniques as there are brachial plexus block, intercostal block and the continuous epidural analgesia using both local anesthetics and spinal opioids. The brachial plexus block in continuous form is absolutely able to prevent pain after operations in the shoulder-arm-region and can be prolonged even for weeks using catheter techniques. The intercostal block on the other hand practically can be performed only as single injection technique being relatively simple however from the technical point of view. The catheter epidural analgesia is the most important method within this group. In comparison to the centrally acting opioids the epidural technique brings some distinct advantages especially in the cardiorespiratory risk case. Choosing between "top up"-technique and continuous infusion of the local anaesthetic depends on the individual circumstances the latter method apparently giving a more steady level of analgesia. The spinal opioid techniques finally gain more and more importance during the last years. They present clear advantages over the local anesthetic methods as there are the long lasting analgesia and the selective blockade of pain not touching motor and sympathetic nerve fibers. A delayed respiratory depression however might be a serious danger showing an incidence of 0,3% in the epidural and some 10% in the subarachnoid route. Aiming to inform the clinician once again about the vast field of possibilities available to make the postoperative course painfree it is hoped that this important task in the postoperative period will be handled with more consequence and effectivity in the future. PMID- 18415569 TI - [Pilot studies with a serotonin agonist (AH 25086 B). Efficacy and tolerance in acute migraine attacks.]. AB - AH 25086 B is a selective agonist of the newly determined 5-HT(1) receptors, which are sited mainly in the intracranial section of the carotid artery. According to experimental studies, the effect of AH 25086 B is decidedly more highly selective than that of ergotamine; the blood flow through the arteriovenous anastomoses of the internal carotid artery is clearly reduced, while the blood flow through the capillaries supplying the brain is increased. With AH 25086 B administered in an infusion rapid abolition of migraine attacks already in progress proved possible. The first 12 patients worldwide to receive this preparation were treated, some on several occasions, for a total of 21 migraine attacks: 7 received one infusion, 3 received two, 1 three, and 1 patient received five infusions. There were some side-effects with some infusions: nausea (18), vomiting (9), and photophobia (19). On average it took 31 min (range 10-60 min) for the headache to be relieved, regardless of the duration of migraine symptoms before the start of treatment. This was not a controlled study, but the results (14 very good, 6 good or satisfactory, in 21 attacks treated) were better than could have been expected by chance. Tolerance of the preparation was good, all side-effects being transitory and mild; with dosages up to 1.6 mug kg(-1) min(-1) no changes were seen in heart rate or blood pressure. PMID- 18415570 TI - [Not Available]. PMID- 18415571 TI - [Not Available]. PMID- 18415572 TI - [Not Available]. PMID- 18415573 TI - [Not Available]. PMID- 18415574 TI - [Not Available]. PMID- 18415575 TI - [Not Available]. PMID- 18415576 TI - [Not Available]. PMID- 18415577 TI - [Not Available]. PMID- 18415579 TI - [Not Available]. PMID- 18415578 TI - [Not Available]. PMID- 18415580 TI - [Not Available]. PMID- 18415581 TI - [Not Available]. PMID- 18415582 TI - [Not Available]. PMID- 18415583 TI - [Not Available]. PMID- 18415584 TI - [Not Available]. PMID- 18415585 TI - [Not Available]. PMID- 18415587 TI - [Not Available]. AB - Non-opioide analgesics have different pharmacological effects and different profiles of their risks. The life-threatening side effects i.e. agranulocytosis after administration of Dipyrone (Metamizol) are rare, but the must be known and cautions must be considered. On the other hand information must be given about the severe side effects following the not-recommended use i.e. end-stage renal disease after misuse of non-opioid analgesics, because these diseases are preventable. PMID- 18415586 TI - [The role of morphine-6-glucuronide during longterm administration of morphine.]. AB - It has been shown that an active metabolite- morphine-6-glucuronide-is formed in man after administration of morphine. In spite of the chemical variation, the glucuronide still possesses high affinity to opioid receptors and the polarity of the glucuronide does obviously not exclude penetration of the blood-brain barrier. After peroral administration of morphine the concentration of the metabolite rises and exceeds within less than 30 minutes the concentration of the parent compound. The affinity profile of morphine-6-glucuronide as measured by binding experiments in membrane homogenates deviates from that of the parent compound. It can not be excluded at the moment that morphine-6-glucuronide will prove superior to morphine for long term treatment, since the kinetic behaviour excludes sharp undulations of the plasma concentration and probably of the concentration at receptor sites in the brain. It is speculated that this type of pharmacokinetic behaviour will reduce tolerance phenomena and abuse liability. PMID- 18415588 TI - [Not Available]. PMID- 18415589 TI - [Not Available]. PMID- 18415590 TI - [Not Available]. PMID- 18415591 TI - [Not Available]. PMID- 18415592 TI - [Not Available]. PMID- 18415593 TI - [Not Available]. PMID- 18415594 TI - [Not Available]. PMID- 18415595 TI - Drug therapy of pain in cancer. AB - CONCLUSION: Most pain in cancer should be easily relieved because it responds in a predictable way to opioid analgesic drugs. Some pains do not respond so well but can usually be ameliorated by the judicious use of adjuvant analgesics, non drug measures, and the active involvement of the multidisciplinary team. PMID- 18415596 TI - [Not Available]. PMID- 18415597 TI - [Not Available]. PMID- 18415599 TI - [Not Available]. PMID- 18415598 TI - [Not Available]. PMID- 18415600 TI - [Not Available]. PMID- 18415601 TI - [Relaxation and biofeedback in the treatment of chronic headache in children and adolescents. A status report.]. AB - The first studies on psychological treatment of pediatric headache appeared about 10 years ago; most of them were published in Anglo-American journals. This review focuses on relaxation training and biofeedback (EMG and hand temperature feedback) as the psychological interventions most often used in pediatric migraine and tension headaches. The results of randomized group studies, which were mostly well-controlled, on the efficacy of relaxation therapy (n=12) now allow a rather optimistic prognosis on the feasibility of this intervention. Most studies demonstrate clinically relevant reductions of headache frequency after training. Biofeedback studies are methodologically less well controlled and although positive effects have been observed, e.g., in single case studies, the relative usefulness of biofeedback has yet to be determined. Deficits in research on the psychological treatment of headache in children and adolescents are described, new research issues are discussed and recommendations for more systematic research are given. PMID- 18415603 TI - [Not Available]. PMID- 18415604 TI - [Pain and functional disturbances in the sacroiliac joint in disc herniation of the lower lumbar spine.]. AB - Back and leg pain in patients with lumbar disc herniation can be caused by various mechanisms. In addition to nerve root compression, functional alterations in the sacroiliac joint, facet joint or the iliolumbar and sacrotuberal ligaments can produce "pseudoradicular" lower back syndrome. The following study attempts to show whether or not pain and functional alterations in the sacroiliac joint (SIJ) correlate with herniations revealed by computed tomography (CT). The study also attempts to determine the correlation between pain and functional changes of the SIJ and the size and level of the disc herniation. Fifty patients with monosegmental disc herniations revealed by CT who showed no signs of bone or soft tissue alterations were included in this study. The average duration of the patients' complaints of leg or back pain was 5.7 years. Ninety-six percent of these patients had received conservative treatment before admission to our hospital. All patients were compared to a control group consisting of 16 healthy subjects of comparable age. All patients underwent a comprehensive functional, neurologic and radiologic examination. The CTs were analyzed by a standardized three-dimensional method. All of the 50 patients had sciatica complaints and a disc herniation revealed by CT. In two cases hemiation of the L3-4 disc was demonstrated, in 14 cases L4-5 disc herniation and in 34 cases a L5-S1 disc herniation. In contrast to the control group of 15 healthy subjects, the patients showed a significant number of functional disorders upon examination. In 84% of all patients, movement of the SIJ was restricted. Painful palpation of the symphysis was demonstrated in 46% of all cases. Thirty-five percent of patients with herniation of L4-5 disc demonstrated SIJ tenderness as opposed to 65% of the patients with herniation of the L5-S1 disc. This SIJ tenderness did not correlate with motion of the SIJ. In addition, SIJ motion and frequency of sensory dysfunction showed no correlation with the size of the disc herniation. Paralysis and loss of reflexes showed a positive correlation with the increasing size of the disc herniation. SIJ tenderness decreased as the size of the herniation increased. Dysfunction of the ipsilateral SIJ is explained by increased muscular tone caused by irritation of the n. sinuvertebralis and its lumbar coupling. Frequency of SIJ tenderness is significantly higher in patients with herniations between L5 and S1. Since the SIJ is innervated by the r. dorsalis of the sacral roots, the increased tenderness can be explained by the change in neurovegetative innervation of the SIJ. Due to the high correlation between lumbar disc herniation and SIJ dysfunction, disc herniation should be considered as a possible cause of sacroiliac-joint syndrome. PMID- 18415602 TI - [Systemic clonidine versus opioids in postoperative analgesia-A randomized double blind study.]. AB - INTRODUCTION: alpha(2)-Adrenozeptoragonisten agonists have shown antinociceptive and analgesic effects, which are not antagonized by naloxone. Therefore, the mechanism of action should be independent of opioid receptors. Most studies on this topic have been performed using clonidine. Experimentally the analgesic effect of clonidine can be suppressed by the inhibition of central adrenergic receptors. Furthermore, clonidine has analgesic effects at the spinal level. During recent years numerous studies have shown the analgesic effect of spinally or epidurally administered clonidine in humans. However, only very few studies have investigated the analgesic effect of parenterally administered clonidine in humans. METHODS: After the approval of the local ethical committee had been obtained, 60 patients (ASA I-III, age 18-65 years) scheduled for elective orthopaedic procedures were included in this double-blind randomized study. All patients gave their written consent on the day before the operation. Premedication was standardized and involved benzodiazepines. Isoflurane was used as the sole anaesthetic. Postoperatively the pain level of the patients was controlled by a visual analogue scale (VAS 0-10). When the VAS reached at least 5 and the patients requested an analgesic, they were randomly assigned to either the morphine, tramadol or clonidine group. Twenty patients received 5 mg morphine i.v., 20 patients received 50 mg tramadol and 20 patients received 150 clonidine i.v. If the analgesic effect was insufficient, the above-mentioned dosage was repeated after 30 min. The therapy was classified as a failure if no sufficent analgesia could be achieved within 60 min. These patients received 7.5 mg piritramide i.v. VAS and sedation were measured at 10-min intervals during the 1 st h and at 15-min intervals during the following 2h. Heart rate, blood pressure and oxygen saturation were measured at 5-min intervals during the whole study period. Statistical analysis of the data was performed by ANOVA, Wilcoxon test, Student'st-test and chi-square test using a level of significance ofP<0.05. RESULTS: All groups were comparable as regards their basic clinical parameters. Morphine, tramadol and clonidine significantly reduced the VAS within 20 min. During the whole study period the analgesic effect of clonidine was comparable with that of morphine and tramadol. No significant differences were observed in the number of repetitions after 30 min or in the failure rate. After 2 h sedation was significantly higher in the morphine group. No clinically relevant cardiovascular or respiratory side-effects were observed in any of the patients. DISCUSSION: In our study the analgesic effect of 150 mug clonidine i.v. was equivalent to that of 5 mg morphine i.v. and 50 mg tramadol. Our results in humans confirm the dosage relationship of 1ratio30 found by Eisenach in sheep. Further studies on the use of parenteral clonidine for postoperative analgesia seem to be warranted. PMID- 18415605 TI - [The so-called ischaemia test-a new diagnostic tool for reflex sympathetic dystrophy.]. AB - Thirty patients suffering from reflex sympathetic dystrophy accompanied by continuous spontaneous pain of the affected distal limb were investigated. The aim of the study was to determine whether a suprasystolic compression (cuff) fixed at the distal part of the affected limb and interrupting the distal circulation would influence the pain. In most cases, this so-called ischaemia test led to a substantial suppression of pain, which occurred during the first or second minute after the cuff had been applied. Therefore, this test can be used as a new diagnostic tool to support the clinical diagnosis of reflex sympathetic dystrophy. PMID- 18415606 TI - [Postoperative pain: patient's self-report versus observer's rating.]. AB - In 60 women undergoing vaginal hysterectomy, a total of 420 pain evaluations of postoperative pain intensity were performed by an observer and the patients. Pain intensity was rated by the observer on a visual analogue scale. The patients themselves evaluated their pain on a visual analogue scale and on a 101-point numerical rating scale. There was good agreement between pain intensity on the visual analogue scale and the 101-point numerical rating scale in the patients' self-assessment (r (2)=0.74;y=0.81x+11.4). The pain intensities determined by the observer on the visual analogue scale were markedly lower, on average than the patients' self-report (P<0.0001). The pain intensities rated by the observer on the visual analogue scale were on averange 37.7% of the patients' self assessments on the visual analogue scale. The correlation between patients' self assessments and observers' ratings was poor (r (2)=0.28;y=0.66x+31.3). There was also no clear correlation between pain intensity and heart rate or arterial blood pressure. A reliable assessment of pain intensity can only be performed by patients' self-assessment and not by observers' ratings. PMID- 18415607 TI - [Revision of the legal prescription requirements for strong opioids in the federal republic of Germany-an initiative of the German association for the study of pain.]. AB - In the Federal Republic of Germany, strict regulations complicate the prescription of strong opioids to patients suffering from severe pain. In summer 1991 the German Association for the Study of Pain launched an initiative directed at simplification of the prescription requirements for such analgesics and submitted the suggestions to the ministries responsible. In the meantime a draft bill has been presented by the Ministry of Health, with due consideration for the main recommendations. For example, substantial increases in the permitted quantities of the individual narcotic analgesics and the prescription periods are planned. Moreover, the prescriptions are to be simpler to make out and certain exemptions have been specified by law. Acceptance of the bill by the political committees and its realization are expected in autumn 1992. PMID- 18415609 TI - [Not Available]. PMID- 18415608 TI - [Diagnostic considerations in muscle pain.]. AB - Although the greatest part of the human body is composed of muscle, diseases of the muscle, such as muscular dystrophies and inflamatory or metabolic myopathies, occur invery few patients. On the other hand, myalgia is one of the most common symptoms in routine clinical medicine. This is problematic, because muscular pain can be caused by many different physical and psychiatric diseases. In order to avoid unecessary and expensive laboratory tests a careful examination of clinical symptoms and signs is necessary. PMID- 18415610 TI - [Not Available]. PMID- 18415611 TI - [Not Available]. AB - Different therapeutic modalities are available for the treatment of rheumatic pain. The most important one, besides physiotherapy, is medication with analgesics and adjuvant drugs. Analgesics are given orally and by a stepwise approach in keeping with the principles of cancer pain therapy. In the first step nonopioid analgesics are prescribed, especially non-steroid anti-inflammatory drugs (NSAID) if pain is caused by inflammation. Other nonopioid analgesics, which can be used as alternatives for patients with non-inflammatory pain, are metamizol and paracetamol. Weak or even strong opioids must be administered to patients with rheumatic diseases when pain relief is insufficient or side-effects occur during medication with non-opioids. Long-term treatment of rheumatic pain even with strong opioids such as oral morphine involves only a small risk of severe side-effects such as respiratory depression or the development of tolerance and drug abuse. Patients often suffer from constipation, nausea and vomiting, but these side-effects can be treated with laxatives and antiemetic drugs. There is no reason to differentiate between opioid medication in a cancer patient with pain and in a patient with "non-malignant" rheumatic pain. Centrally acting muscle relaxants may be helpful as adjuvant medication in patients with myalgia for example, and tricyclic antidepressants can also be beneficial, especially in neuropathic pain and for patients with psychiatric distress associated with pain. PMID- 18415612 TI - [Pumps and ports for analgesic therapy.]. AB - At one of the symposia at the 16th congress of the German Society for the Study of Pain in Berlin in 1991 an update on the use of pumps and ports in pain treatment was presented. This article tries to focus on some of the conclusions of this meeding. (1) To avoid neurologial damage only analgesic substances that have been tested in animals and with which we have adequate clinical experience, e.g. opiates, clonidine and baclofen, should be used for spinal anaesthesia. (2) The increasing number of manufactures of pumps, ports, catheters and puncture needles should pay more attention to safety and reliability of their products to avoid technical complications and should try to achieve compatibility between the different accessories such as catheters, needles and connecting pieces. (3) The mode of application (spinal versus epidural, pump versus port or externalized catheter) depends on the predicted period of treatment (spinal application for long-term treatment requires more hygienic precautions) and on differences in the individual care of the patient (4). Spinal opiates for benign pain (in case of failure of less invasive pain regiments) have been tested successfully, but the follow-up is not yet long enough to allow recommendation of this therapy for general use. (5) Local anaesthetics are useful for short-term use. The addition of local anaesthetics for continuous low-flow infusion of opiates requires further comparative studies. (6) Spinal baclofen is effective against pain induced by muscle spasms but not against non-spasticity-related pain syndrome. PMID- 18415613 TI - [How useful is the combination of B vitamins and analgesic agents?]. AB - Vitamins of the B group have long been used to treat neuropathies of different origins and the accompanying pain. A combination of the vitamins B(1), B(6), and B(12) prevents the slowing of impulse conduction produced by tetanic stimulation in diabetic mice. In patients suffering from diabetic neuropathy, B vitamins alleviate pain in the upper extremities. Thermosensitivity is restored by B vitamins in the upper but not in the lower extremities. It has recently also been reported that a combination of the vitamins B1, B6, and B12 has analgesic properties in non-neuropathic conditions. In animal experiments, B vitamins diminish nociceptive responses in spinal and thalamic neurones and potentiate the antinociceptive effect of analgesic agents. Similarly, B vitamins potentiate the therapeutic effect of diclofenac in patients suffering from acute low back pain. PMID- 18415614 TI - [Prevalence and characteristics of neuropathic pain in malignant disease.]. AB - Neuropathic pain is one of the problem areas in the management of cancer pain. In a retrospective study, prevalence and characteristics of neuropathic pain in 1318 cancer patients attending a pain clinic were examined. Of the patients, 135 suffered from neuropathic, 285 from neuropathic and nociceptive, 890 from nociceptive and 8 from unknown pain conditions. Among the patients with neuropathic pain 62% rated the pain intensity as very sincere; this was so in 48% of those with nociceptive pain. Neuropathic pain was caused by direct tumour involvement (nerve compression or infiltration) in 71%, by oncological treatment (surgery, chemotherapy, radiation) in 15%, by debilitating disease (e.g. herpes zoster) in 6% and by factors unrelated to cancer or its treatment in 8% of the patients. Of 110 clinically analysed neuropathic pain conditions, 44% were neuralgic, 31% radicular, 13% sympathically maintained, and 10% caused by deafferentiation, while in 3% the nature was unknown. To evaluate the efficacy of cancer pain treatment, nocicepetive pain has to be differentiated from neuropathic pain. In addition to this, neuropathic pain has to be divided into subgroups. PMID- 18415616 TI - [Not Available]. PMID- 18415615 TI - [Epidural steroid injections in the treatment of back pain. A retrospective and prospective analysis.]. AB - Epidural steroid injections are frequently used in the conservative treatment of backache, although they are still subject to critical discussion. Relief of pain is attributed to the anti-inflammatory effect of the steroid. During a 3-year period, 53 patients with back pain or differing aetiology were treated with one or more epidural injections of 14 mg betamethasone (2 ml Celestan) in a prospective and retrospective fashion. Patients were followed up for 1 year. Immediately after the injection 68% free of pain. Freedom from pain or improvement totally free of pain. Freedom from pain or improvement was reported by 39% of patients after 2 weeks and by 31% after 6 months to 1 year. Patients with acute pain (up to 6 months) responded better than patients with chronic symptoms. No significant correlations were detected between response and other characteristics, e.g. age, sex, number of injections, type of pain, intensity of pain, or psychological overlay. For patients with acute pain epidural steroid injections seem to be a safe, appropriate and promising procedure. PMID- 18415617 TI - [Chronic low back pain and life events.]. AB - In a retrospective, controlled clinical study the life events of 35 patients suffering from chronic low back pain (LBP) and a matched sample of 23 patients with neurotic depression (ICD 300.4) were investigated. The pain patients formed two groups: 19 patients with definite organic diagnosis (IASP code 530.96) and 16 without (adequate) organic lesion (IASP code 510.99). Somatic diseases (other than LBP), injuries and operations, as well as psychic trauma (feelings of shame, narcissistic traumatisations and object losses) were defined and counted as documented in the patient's histories. Their distribution over the period between the 10th year before onset of LBP resp. depression and the 10th year after onset was calculated. As a result all patients have to cope with a maximum of stressful life events in the initial year of their disease. Those patients, who fell ill younger than 30 years old, are confronted with an increased number of stressful life events even during several years before the onset. In all groups significant more stress is experienced after illness onset compared with the time before onset. LBP patients without organic findings experience more narcissistic traumatisations than the other two groups in the initial year and later on. In contrast depressive patients suffer from more other diseases, injuries and have undergone more operations than pain patients throughout the whole time investigated. Object losses occur equally often in all groups, apart from the initial year, when depressive patients have to cope with even more losses than the others. These results are discussed considering the development of chronic pain syndromes, the influence of age and their consequences for models of illness. There is convincing evidence, that physical injury is neither a necessary nor a sufficient condition for the development of chronic pain and that chronic pain is in essence an emotional disease based on unresolved unconscious conflicts requiring psychotherapy. PMID- 18415618 TI - [Progressive relaxation and EMG biofeedback in the treatment of chronic headache in children. Results of an explorative study.]. AB - A group study on the comparative efficacy of EMG biofeedback and progressive relaxation is presented. Sixteen children aged between 8 and 14 years with chronic tension headache and combined headache participated in the study. Six sessions of relaxation training and 12 (shorter) biofeedback sessions were held with each child. Both treatments had excellent results, which were apparent directly after training. All but one child benefited to a clinically significant extent from the treatment, with a reduction of more than 50% in headache frequency. Other variables indicate further positive effects of treatment (e.g., medication consumption, absence from school). After 6 months of follow-up the children treated by relaxation had achieved event further reductions in headache activity. Suggestions for further improvement in the clinical and economic efficiency of treatment formats are presented, and perspectives for future research are discussed. PMID- 18415619 TI - [Acupuncture for the treatment of pain-a prospective study.]. AB - Acupuncture is increasingly being used in Western medicine. In the present study the effect of acupuncture treatment was examined in 60 patients with chronic pain. The long-term success after 6 months was evaluated in 45 patients. Calculated on the basis of an index of the frequency of pain, its duration and intensity, a global success rate of 64% was found directly after the therapy and of 69% 6 months later. Furthermore, we examined to what extent the success of the therapy was dependent on several variables. It appeared that the consumption of analgesics could be reduced significantly in patients in whom the treatment was successful. All patients had been treated before by several methods. Because of the small number of side-effects and the patients' high degree of acceptance as well as the success of the treatment, we consider acupuncture an effective method in the treatment of chronic pain. PMID- 18415620 TI - [The efficacy of classic homeopathic therapy in chronic headache. Study protocol.]. AB - The efficacy of classic homeopathic therapy is scientifically not well proven. Few of the studies available are acceptable from a scientific point of view. In this paper we will describe a study protocol for a trial of classic homeopathy in chronic headache, which not only is in accordance with currently accepted scientific research standards but also accounts for the special needs of homeopathic therapy. The prescribing practioners are allowed to use any homeopathic drug at any potency or dosage, this enables them to ful fill the homeopathic principle of individuality. At the same time, the study is strictly controlled, randomized and double blind. The prescribing physician sends the homeopathic medication selected for a patient to a notary public, who either forwards it to the patient or substitute placebo, according to the randomization plan. Hence, the trial is not testing a specific drug, but the rationale of individual homeopathic drug selection and the efficacy of the selected drugs in headache patients in general. Patients suffering from chronic headaches for at least 1 year and with headaches at least once a week on average are eligible for the study. Exclusion criteria follow the generally accepted standards and account for the special needs of homeopathic therapy. Data are to be recorded in a patient's diary containing the outcome variables occurrence, duration, intensity of headaches and use of analgesic drugs. After a baseline period of at least 6 weeks the first consultation will take place, with the proper remedy selected and sent to the patient via the notary. After 6 weeks a first follow-up will allow the physician to modify the treatment if necessary. After another 6 weeks the final examination will take place. The duration of the homeopathic treatment, then, is 12 weeks. A total of 100 patients in two groups of 50 are to be treated in the study. The study started in the later part of 1991, and is scheduled to last for 2 1/2 years. We expect a critical discussion of the results from conventional medicine or from homeopathy, depending on the outcome of the study. The study protocol is being published in advance to enable the reviewers of the study to check the original study design and the a priori hypotheses adopted. PMID- 18415621 TI - [Treatment of trigeminal neuralgia with vincristine iontophoresis. A clinical pilot study.]. AB - Treatment of chronic pain disease is a scientific and clinical challenge encountered in all branches of medicine. Essential trigeminal neuralgia and chronic pain situations are noted for their exceptionally marked severity and also for their psychic and social consequences. It is not rare for a primarily somatic pain syndrome to develop into a painful disease in its own right, which is highly refractory to treatment. In transcutaneous iontophoresis of vincristine the neurotoxic effect of the alkaloid is employed locally and in a controlled manner. Iontophoresis of a microtubule inhibitor (vincristine, vinblastin) to the skin induces transganglionic degenerative atrophy of the central terminals of primary sensory neurons. Vincristine iontophoresis was applied in 33 patients hitherto unsuccessfully treated with various other methods. In 78% of the cases, attenuation of the pain was achieved. This noninvasive therapy proved to be free of side effects. PMID- 18415622 TI - [Not Available]. PMID- 18415623 TI - [Anticonvulsant agents in neuralgic pain.]. AB - The anticonvulsants, carbamazepine, clonazepam, phenytoin, and valproic acid are capable of depressing attacks of shooting pain in neuralgia. Shooting pain is perceived in trigeminal, intercostal, and other neuralgias, as a consequence of infectious diseases such as herpes zoster, and in the course of polyneuropathies of various causes. It is due to injury of nociceptive afferents, which generate bursts of activity in response to appropriate environmental changes. The anticonvulsant agents have no analgesic property per se, so that background pain remains unchanged. The depression of shooting pain results from the anticonvulsant action of the compounds. Both carbamazepine and phenytoin block synaptic transmission of neuronal hyperactivity by a direct depressant action that includes reduction of sodium conductance and by activation of inhibitory control. Clonazepam and valproic acid act by enhancing GABA-mediated inhibition of synaptic transmission. Carbamazepine is by far the most widely used compound; phenytoin, clonazepam, and valproic acid are not so popular because of their side effects. PMID- 18415624 TI - [Not Available]. PMID- 18415625 TI - [Not Available]. PMID- 18415626 TI - [Some considerations about the treatment of trigeminal neuralgia with intrathecal baclofen.]. AB - A case history of therapy resistent trigeminal neuralgia was presented by Wechsung and Muller. This patient was previously treated without any succes with glycerol injections, "electrocauterisation", a Janetta operation and 2 reexplorations of the posterior fossa. Attempts with oral carbamazepine and baclofen were unsuccesful too. The authors did not try out the combination of carbamazepine and baclofen, which is recommended by Fromm and Terrence. When conservative treatment fails, surgical procedures are indicated. The most succesful surgical procedure is the radiofrequency thermal lesion of the Gasserian ganglion (Sweet). Data from the literature from 8500 patients and own data (from over 600 patients) show that therapy resistance is very infrequent (0.5%). The costs of clinical implantation of a pump with intrathecally administered baclofen are 10-15 times higher than ambulant radiofrequency lesioning. PMID- 18415627 TI - [Subcutaneous CO(2) insufflation therapy.]. PMID- 18415628 TI - [Not Available]. PMID- 18415629 TI - [Not Available]. PMID- 18415630 TI - [Treatment of a high-grade pain syndrome in partial paralysis by series of paravasalic sympathetic nerve blockade.]. AB - Because of distinct pain symptoms in the presence of an ascending partial paralysis-Brown-Sequard Syndrome-from T 5/6 upward in a 56-year-old patient, therapy with paravasalic injections of bupivacaine 0.125% in the area of the femoral artery (right) was given. The patient kept a pain diary with entries every 2 h for 61 weeks. Evaluation of the documentation demonstrates the success of the therapy and shows the possibilities of different kinds of evaluation of well-documented pain data. PMID- 18415631 TI - [Not Available]. PMID- 18415632 TI - [Not Available]. PMID- 18415633 TI - [Analgesia with opioids in the paediatric patient.]. AB - Opioids have been used for analgesia in nearly all civilizations. In paediatrics their use has become widely accepted for combating severe pain, especially postoperative pain and tumour pain. Receptors in the central nervous system are the best known sites of action of opioids, but the existence of peripheral receptors is also probable. The action depends on whether the opioid is more agonist or antagonist and on the peculiarities of physiology in childhood: in the small child a hyperdynamic blood circulation makes resorption faster, and in newborn and premature infants distribution and excretion are influenced by the different composition of the body and the immaturity of liver and kidney. The best known opioid is morphine, and it is the reference substance with which all other opioids are compared. Fentanyl has been used even for the smallest ventilated prematures in recent times, as it is easy to manage and has an early onset of action. Its depressant action on the respiratory centre is an advantage when attempts of spontaneous breathing make mechanical ventilation difficult. Obstinate constipation is the disadvantage of both morphine and fentanyl, and an exacerbation of hyperbilirubinaemia has been seen with fentanyl. Nalbuphine causes a lower degree of respiratory depression. The newer opioids alfentanil and sufentanil have already been used for the relief of paediatric postoperative pain and during mechanical ventilation, but no special advantages of their use are reported. Meperidine has been favoured especially for postoperative pain, although it appears to have no advantages over morphine. Its active metabolite normeperidine may accumulate and cause seizures; meperidine should not be used in prematures or in children with renal dysfunction. There are few publications on the use of piritramide in paediatric pain. Tramadol is widely used for emergencies, as it has the least sedative action; but it has disadvantages in causing nausea and vomiting. Codeine is widely used for its antitussive action. While the necessity of good analgesia for even the smallest infant cannot be overstated, the opioid used must be carefully selected with reference to the age of the child and the pain to be controlled. PMID- 18415634 TI - [Does anginal pain influence the medical care-seeking behavior of patients in the prodromal phase prior to an acute myocardial infarction. Results of a post infarction late potential study.]. AB - The aim of the study was to determine the proportion of high-risk patients who received appropriate antianginal therapy in the prodromal phase prior to a myocardial infarction, as an indicator of medical care seeking behavior. To this end, 606 male infarct patients aged 29-65 years were retrospectively interviewed 17-21 days after acute myocardial infarct. It was found that 77% of all patients (465/606) suffered from anginal pain, but only 32% of the patients with angina pectoris were receiving antianginal therapy in the prodromal period before acute myocardial infarction. Patients not taking medication were significantly younger than those with antianginal medication; they were more often smokers; they were less often suffering from high blood pressure; they expressed more pronounced nonacceptance of the risk; their history of anginal pain was significantly shorter; and they belonged more often to the patient group with a first myocardial infarction. In stepwise logistic regression analysis, high blood pressure, older age and exhaustion were found to be associated with medical treatment before infarction in the patient group with first myocardial infarction. In patients with recurrent infarction, continued smoking and denial of the risk remained predictive of nonmedication. PMID- 18415635 TI - [Is intravenous lidocaine infusion suitable for postoperative pain management?]. AB - Intravenous administration of local anaesthetics has repeatedly been recommended for the treatment of chronic pain. Some authors have also reported on their use in postoperative pain management. However, most of these publications are case reports or refer to rather old studies or investigations based on study designs that fail to meet present scientific standards. We therefore performed a randomized prospective, double-blind study in 40 patients undergoing elective tonsillectomy, 20 of whom received an infusion of lidocaine at a dose of 1.5 mg/kg body weight (over 10 min) 30 min before the beginning of surgery, followed by 2 mg/kg body weight per h over 6 h and 0.5 mg/kg body weight per h for another 18 h. The patients in the control group received identical volumes of 0.9% NaCl solution. Mean lidocaine plasma concentrations determined 30 min and 3, 6, and 24 h after the beginning of surgery ranged between 2.29 and 0.58 mug/ml. Postoperative pain evaluation on the visual analogue scale and the 101-point numerical rating scale did not reveal,significantly lower pain scores in the lidocaine group than in the control group. During the first 24 h after surgery 12 patients in the lidocaine group required a total of 550 mg meperidine in addition, while 8 patients in the control group required a total of 300 mg meperidine. The postoperative meperidine consumption was not significantly diffent between the lidocaine group and the control group. Intravenous lidocaine infusion did not significantly reduce postoperative pain after tonsillectomy in the dosage used. PMID- 18415636 TI - [Pain and functional disturbances of the lumbar spine in lumbar herniated discs.]. AB - In this study we attempted to explore the correlation between lumbar disc herniation and functional disorders of the lumbar spine. Fifty patients with lumbar disc herniation proven by computed tomography underwent a comprehensive functional, neurological and radiological examination. All patients were compared to a control group consisting of 16 healthy subjects of comparable age. Only patients without signs of bone or soft tissue alterations or pregnancy at the time of examination were included into the study. Herniations of the L4-5 disc showed a dysfunction in the same segment in 64% of the cases. There was also a correlation between this segmental dysfunction and pain in the sacrotuberal and iliolumbar ligaments. All patients with segmental dysfunction felt pain in the dorsal ligaments. If there was no segmental dysfunction pain in the dorsal ligaments was encountered just as often as in the control group. Herniations of the L5-S1 disc had a dysfunction in the same segment in only 12% of the cases, but in 35% there was dysfunction of the L4-5 motion segment. In this group pain in the dorsal ligaments did not correlate with segmental movements. Frequency of ligamental pain in L4-5 herniations was equal to that in L5-S1 herniations. With increasing size of the disc herniation, the frequency of segmental dysfunction, paralysis and loss of reflexes also increased, but the pain in the dorsal ligament decreased.Segmental dysfunction is explained by increased muscular tone being provoked by irritation of the sinuvertebral nerve. The differences between L4-5 and L5-S1 movements are probably due to the different functional anatomy of these segments. Ligamental pain may be explained by the fact that these ligaments have the same insertion and the muscles have increased in tone. PMID- 18415637 TI - [Prescription of opiates. Analysis of outpatient care of pain patients insured with the AOK.]. AB - INTRODUCTION: The efficacy of morphine and other opioids in the treatment of severe pain, especially cancer pain, is undoubted. If opioids are used according to the WHO guidelines no severe side effects or psychological dependence develop. In Germany the use of strong opioids is controlled by law and requires a special prescription with offical forms and difficult regulations. METHODS: In Bochum, a city in Germany with about 400000 inhabitants, we analysed the treatment of outpatients with strong opioids in the last 3 months of 1989 and the first 3 months of 1990. RESULTS: Only 55 of the 92842 people insured with the Bochum branch of the AOK, a large health insurance company, received a prescription for strong opioids in these two quarters (0.059% of all members): 42 of these patients had a cancer disease and 13 a disease of nonmalignant origin. In all, these patients received 504 prescriptions for strong opioids (0.078% of the total number of prescriptions). Most of the patients received only one to five prescriptions. Only two patients with cancer pain received more than 20 prescriptions. Of all physicians in Bochum only 10.88% prescribed strong opioids. CONCLUSIONS: The prescription patterns we found demonstrate inadequate medical care of patients with severe pain. Continuous pain therapy with strong opioids was found in only a few cases. The main reason for this situation is the law pertaining the use of opioid drugs, with its difficult and restrictive regulations. This law cannot stop the development of increasing illegal drug consumption, but obstructs the medical care of patients with severe pain. PMID- 18415638 TI - [Continuous subcutaneous application of analgesics in a patient suffering from a head and neck tumor.]. AB - In the pain treatment of patients with incurable head and neck tumors it may be difficult to provide adequate oral drug therapy in the advanced stages. In these cases continuous subcutaneous application of morphine by means of external infusors is an alternative. An example of the method of continuous subcutaneous application of analgesics in pain therapy is shown in a patient suffering from a chondrosarcoma of the maxilla. Because oral pharmacotherapy provided insufficient analgesia in the final stage of the tumor disease, it was replaced by continuous subcutaneous application of morphine hydrochloride. A flexible cannula was inserted into the upper arm and connected to an external portable infusor device. Because of growing tumor infiltration, we progressively increased the morphine dosage. Nevertheless, continous subcutaneous application of droperidol, dexamethasone and clonidine was also needed to achieve much better analgesia. The technical simplicity of this safe and effective treatment make it suitable for outpatient care in tumor clinics. PMID- 18415639 TI - [Not Available]. PMID- 18415640 TI - [Can caffeine be regarded as a proven adjuvant to minor analgesics?]. PMID- 18415641 TI - [Drug treatment for pain in children and elderly persons.]. AB - The subject of treatment of pain in children and elderly patients has been neglected to date. This symposium deals with the problems of pain treatment in these two age groups and shows the importance of adequate therapy to improve their quality of life. PMID- 18415642 TI - [Not Available]. PMID- 18415644 TI - [Not Available]. PMID- 18415643 TI - [Not Available]. PMID- 18415645 TI - [Not Available]. PMID- 18415646 TI - [Not Available]. PMID- 18415647 TI - [Not Available]. PMID- 18415648 TI - Quantitative kinetic analysis of lung nodules using the temporal subtraction technique in dynamic chest radiographies performed with a flat panel detector. AB - Early detection and treatment of lung cancer is one of the most effective means of reducing cancer mortality, and to this end, chest X-ray radiography has been widely used as a screening method. A related technique based on the development of computer analysis and a flat panel detector (FPD) has enabled the functional evaluation of respiratory kinetics in the chest and is expected to be introduced into clinical practice in the near future. In this study, we developed a computer analysis algorithm to detect lung nodules and to evaluate quantitative kinetics. Breathing chest radiographs obtained by modified FPD and breath synchronization utilizing diaphragmatic analysis of vector movement were converted into four static images by sequential temporal subtraction processing, morphological enhancement processing, kinetic visualization processing, and lung region detection processing. An artificial neural network analyzed these density patterns to detect the true nodules and draw their kinetic tracks. Both the algorithm performance and the evaluation of clinical effectiveness of seven normal patients and simulated nodules showed sufficient detecting capability and kinetic imaging function without significant differences. Our technique can quantitatively evaluate the kinetic range of nodules and is effective in detecting a nodule on a breathing chest radiograph. Moreover, the application of this technique is expected to extend computer-aided diagnosis systems and facilitate the development of an automatic planning system for radiation therapy. PMID- 18415649 TI - Implementation of quality measures to reduce surgical site infection in colorectal patients. AB - PURPOSE: The goal of this study was to determine the rate of surgical site infection and compliance with process measures designed to prevent infection in a defined population of patients undergoing colorectal operations. METHODS: A task force consisting of surgeons, hospital infection control personnel, anesthesiologists, and nurses was convened to enforce the use of process measures to prevent infections. We monitored antibiotic selection, dosage, timing, redosing and discontinuation, hair removal technique, intraoperative and postoperative body temperature, and perioperative glucose control for 12 months by using electronic medical records. Patients underwent a minimum of 30 days of postoperative follow-up and the attending surgeon diagnosed infections. RESULTS: Between April 2006 and March 2007, 298 patients underwent abdominal colorectal operations. The overall infection rate was 20 percent for colon procedures and 11 percent for small-bowel procedures. Compliance for most process measures improved from the first to the fourth quarter, and during the final quarter, correct antibiotic dose and hair removal with clippers exceeded 90 percent. CONCLUSIONS: The rate of surgical site infection after colorectal surgery is likely to be higher than that reported in national quality improvement programs. Perfect compliance with performance measures may be difficult to attain. PMID- 18415650 TI - Long-term morbidity of sentinel node biopsy versus complete axillary dissection for unilateral breast cancer. AB - BACKGROUND: Postoperative changes after axillary lymph node surgery may significantly alter breast cancer survivors' (BCS) quality of life. Although sentinel lymph node biopsy (SLNB) has less immediate morbidity than axillary lymph node dissection (ALND), its long-term impact on shoulder abduction, arm swelling, and neurosensory changes has not been evaluated. The purpose of this study was to compare long-term morbidity after SLNB or ALND and breast conservation surgery. METHODS: Female BCS who remained free of disease at least 3 years after ALND or SLNB for Stage I-III unilateral breast cancer completed a symptom questionnaire and a brief neurosensory physical examination of the upper arm and axilla (range of motion, arm circumference, and sensation to light touch with cotton and needle). RESULTS: The mean age of the 187 participating BCS was 62 years. At a mean follow-up of 6.6 years after ALND and 4.9 years after SLNB, most BCS had full abduction; only 10 cases (8 ALND, 2 SLNB) had a >/=2 cm proximal and/or distal circumference difference on the ipsilateral side compared with the contralateral side. ALND was associated with a significantly greater likelihood of subjective arm numbness (P < .001), chest or axillary numbness (P < .001), arm or hand swelling (P < .001), and objective neurosensory changes in the posterior axilla, medial and distal upper arm (P < .001). Operative procedure was the only significant predictor of neurosensory changes (P < .001). CONCLUSION: SLNB is associated with significantly less subjective and objective long-term morbidity than ALND. PMID- 18415651 TI - Angiography is indicated for every sentinel bleed after pancreaticoduodenectomy. AB - BACKGROUND: Delayed massive bleeding is one of the leading causes of mortality after pancreaticoduodenectomy (PD) and is often preceded by sentinel bleed. Immediate and accurate diagnosis of sentinel bleed is essential to save patients from a delayed massive hemorrhage. Angiography is probably the procedure of choice for patients with sentinel bleed after PD, as it will localize the bleeding point and provide interventional embolization. The purpose of this study is to test the efficiency of angiography as the initial management for patients with sentinel bleed after pancreaticoduodenectomy. METHODS: The study group consisted of 283 patients who underwent PD from July 2002 to June 2007. Angiography and arterial embolization were performed for every sentinel bleed and detected pseudoaneurysm. Patients (n = 311) from a previous study (July 1996-June 2002) were used as a historical control group. RESULTS: Sentinel bleed was detected in 20 patients in study group. Of these, angiography-detected pseudoaneurysm was evident in seven (35%); all were successfully embolized. Delayed massive hemorrhage occurred in three of 13 patients with sentinel bleed but negative angiography. All three were operated on; one died of uncontrolled bleeding. The number of hemodynamically unstable patients before transfusion, units of transfused packed cells, and bleeding related mortalities were significantly less in study group than the control group. CONCLUSIONS: Institution of angiography for every detected sentinel bleed after PD enabled us to embolize seven pseudoaneurysms before massive hemorrhage. Most importantly, bleeding-related mortality was significantly less than in the absence of angiography. PMID- 18415652 TI - Herbal medicine in the United States: review of efficacy, safety, and regulation: grand rounds at University of California, San Francisco Medical Center. AB - INTRODUCTION: Herbal products have gained increasing popularity in the last decade, and are now used by approximately 20% of the population. Herbal products are complex mixtures of organic chemicals that may come from any raw or processed part of a plant, including leaves, stems, flowers, roots, and seeds. Under the current law, herbs are defined as dietary supplements, and manufacturers can therefore produce, sell, and market herbs without first demonstrating safety and efficacy, as is required for pharmaceutical drugs. Although herbs are often perceived as "natural" and therefore safe, many different side effects have been reported owing to active ingredients, contaminants, or interactions with drugs. RESULTS: Unfortunately, there is limited scientific evidence to establish the safety and efficacy of most herbal products. Of the top 10 herbs, 5 (ginkgo, garlic, St. John's wort, soy, and kava) have scientific evidence suggesting efficacy, but concerns over safety and a consideration of other medical therapies may temper the decision to use these products. CONCLUSIONS: Herbal products are not likely to become an important alternative to standard medical therapies unless there are changes to the regulation, standardization, and funding for research of these products. PMID- 18415653 TI - Development and testing of the Multidimensional Trust in Health Care Systems Scale. AB - OBJECTIVE: To describe the development and psychometric testing of the Multidimensional Trust in Health Care Systems Scale (MTHCSS). METHODS: Scale development occurred in 2 phases. In phase 1, a pilot instrument with 70 items was generated from the review of the trust literature, focus groups, and expert opinion. The 70 items were pilot tested in a sample of 256 students. Exploratory factor analysis was used to derive an orthogonal set of correlated factors. In phase 2, the final scale was administered to 301 primary care patients to assess reliability and validity. Phase 2 participants also completed validated measures of patient-centered care, health locus of control, medication nonadherence, social support, and patient satisfaction. RESULTS: In phase 1, a 17-item scale (MTHCSS) was developed with 10 items measuring trust in health care providers, 4 items measuring trust in health care payers, and 3 items measuring trust in health care institutions. In phase 2, the 17-item MTHCSS had a mean score of 63.0 (SD 8.8); the provider subscale had a mean of 40.0 (SD 6.2); the payers subscale had a mean of 12.8 (SD 3.0); and the institutions subscale had a mean of 10.3 (SD 2.1). Cronbach's alpha for the MTHCSS was 0.89 and 0.92, 0.74, and 0.64 for the 3 subscales. The MTHCSS was significantly correlated with patient-centered care (r = .22 to .62), locus of control-chance (r = .42), medication nonadherence (r = .22), social support (r = .25), and patient satisfaction (r = .67). CONCLUSIONS: The MTHCSS is a valid and reliable instrument for measuring the 3 objects of trust in health care and is correlated with patient-level health outcomes. PMID- 18415654 TI - Herpes simplex encephalitis and subsequent cytomegalovirus encephalitis after chemoradiotherapy for central nervous system lymphoma: a case report and literature review. AB - Neurological complications during the treatment of hematological malignancies have a wide range of causes. Treatment-related leukoencephalopathy has been recognized as a major complication of combined chemotherapy and radiotherapy for central nervous system (CNS) lymphoma, and can complicate the diagnosis of CNS infection. Herein, we present a patient with diffuse large B-cell lymphoma who developed herpes simplex encephalitis (HSE) and subsequent cytomegalovirus encephalitis after chemoradiotherapy for CNS relapse. Although cerebrospinal fluid examination (CSF) showed no significant pleocytosis, brain magnetic resonance imaging and polymerase chain reaction analysis of the CSF were useful in the diagnosis. With a review of the literature on the association between HSE and radiotherapy for CNS malignancies, our case suggests that an awareness of viral encephalitis is important in the differential diagnosis of acute neurologic disturbance during chemoradiotherapy for CNS lymphoma. PMID- 18415655 TI - Efficient retroviral transduction of human B-lymphoid and myeloid progenitors: marked inhibition of their growth by the Pax5 transgene. AB - We applied a coculture system for the genetic manipulation of human B-lymphoid and myeloid progenitor cells using murine bone marrow stromal cell support, and investigated the effects of forced Pax5 expression in both cell types. Cytokine stimulated cord blood CD34+ cells could be transduced at 85% efficiency and 95% cell viability by a single 24-h infection with RD114-pseudotyped retroviral vectors, produced by the packaging cell line Plat-F and bicistronic vector plasmids pMXs-Ig, pMYs-Ig, or pMCs-Ig, encoding EGFP. Infected CD34+ cells were seeded onto HESS-5 cells in the presence of stem cell factor and granulocyte colony-stimulating factor, allowing the extensive production of B progenitors and granulocytic cells. We examined the cell number and CD34, CD33, CD19, and CD20 lambda and kappa expressions by flow cytometry. Ectopic expression of Pax5 in CD34+ cells resulted in small myeloid progenitors coexpressing CD33 and CD19 and inhibited myeloid differentiation. After 6 weeks, the number of Pax5-transduced CD19+ cells was 40-fold lower than that of control cells. However, the expression of CD20 and the kappa/lambda chain on Pax5-transduced CD19+ cells suggests that the Pax5 transgene may not interfere with their differentiation. This report is the first to describe the effects of forced Pax5 expression in human hematopoietic progenitors. PMID- 18415656 TI - Anti-myeloma effect of homoharringtonine with concomitant targeting of the myeloma-promoting molecules, Mcl-1, XIAP, and beta-catenin. AB - Since a variety of cell intrinsic and extrinsic molecular abnormalities cooperatively promote tumor formation in multiple myeloma (MM), therapeutic approaches that concomitantly target more than one molecule are increasingly attractive. We herein demonstrate the anti-myeloma effect of a cephalotaxus alkaloid, homoharringtonine (HHT), an inhibitor of protein synthesis, through the induction of apoptosis. HHT significantly reduced Mcl-1, a crucial protein involved in myeloma cell survival, in all three myeloma cell lines examined, whereas certain BH3-only proteins, such as Bim, Bik, and Puma, remained unchanged following HHT treatment, and their expression levels depended on the cell type. HHT also reduced the levels of c-FLIP(L/S), activated caspase-8, and induced active truncated-Bid. Thus, HHT-induced apoptosis appears to be mediated via both intrinsic and extrinsic apoptosis pathways, and the resultant imbalance between BH3-only proteins and Mcl-1 may be pivotal for apoptosis by HHT. In addition, HHT treatment resulted in reduced levels of beta-catenin and XIAP proteins, which also contribute to disease progression and resistance to chemotherapy in MM. In combination, HHT enhanced the effects of melphalan, bortezomib, and ABT-737. These results suggest that HHT could constitute an attractive option for MM treatment though its ability to simultaneously target multiple tumor-promoting molecules. PMID- 18415657 TI - ESHAP therapy effective in a patient with Langerhans cell sarcoma. AB - We describe the rare case of a 53-year-old woman with systemic involvement of Langerhans cell sarcoma (LCS) who had undergone living-related liver transplantation. We chose the CHOP regimen as first-line chemotherapy, and clinical improvement of LCS was obtained. Intensive care was necessary due to the systemic involvement of LCS and severe infectious diseases. After the third cycle of CHOP therapy, however, disease progression was observed, and we administrated a modified ESHAP regimen (etoposide, carboplatin, cytarabine, methylprednisolone) as second-line therapy. A marked response was obtained after four cycles of this combination chemotherapy. Modified ESHAP may be a very effective combination chemotherapy regimen for LCS. PMID- 18415658 TI - HERG K+ channel expression in CD34+/CD38-/CD123(high) cells and primary leukemia cells and analysis of its regulation in leukemia cells. AB - The human ether-a-go-go-related (herg) gene encoding K+ channels (HERG) belongs to an evolutionarily conserved multigene family of voltage activated K+ channels. The functional properties of HERG K+ channels are complex and their contribution to the repolarization of the cardiac action potential are well understood. Recent studies revealed that HERG K+ channels are preferentially expressed in different histogenesis of tumor cells. Leukemia is a cancer that originates in the bone marrow hematopoietic stem cells (HSCs). Leukemia stem cells (LSCs) are critical in the perpetuation of the disease. A better understanding of LSCs and molecular biology will allow the design of more effective therapies. We report in this study that herg was expressed in CD34+/CD38-/ CD123(high) LSCs but not expressed in normal bone marrow CD34+/CD38- HSCs. In addtion, herg is also expressed in leukemia cell lines K562 and HL-60 and almost all the primary leukemia cells whereas not in the normal bone marrow cells. In addition, the expression of herg mRNA was not associated with the clinical and cytogenetic feature of leukemia. Moreover, HERG K+ channels can regulate leukemia cells proliferation and cell cycle. These data provide evidence for the oncogenic potential of HERG K+ channels and it may be a novel, potential pharmacological target for leukemia therapy in the future. PMID- 18415660 TI - Evaluation of sensitivity and specificity of doublecortin immunostatining for the detection of infiltrating glioma cells. AB - Diffuse gliomas are highly infiltrative intracranial tumors, but there are few useful markers for detecting infiltrating glioma cells in the surrounding brain tissue. Doublecortin (DCX) is a microtubule-associated protein (MAP) that plays a crucial role in neuroblast migration. It was recently demonstrated that DCX is preferentially expressed in invasive gliomas. However, the sensitivity and specificity of DCX as a marker for infiltrating glioma cells have not been fully evaluated. We immunohistochemically analyzed the expression pattern of DCX in human gliomas and compared it with that of MAP-2e, another marker for infiltrating glioma cells. We found that DCX was expressed specifically in infiltrating gliomas, but not in reactive, existing glia. Not all our cases exhibited stronger immunoreactivity to DCX at the invasive margin than at the core mass. The level of DCX expression was more variable from case to case than that of MAP-2e. For the identification of infiltrating glioma cells, DCX was thus more specific than MAP-2e whereas MAP-2e was more sensitive than DCX. DCX immunostaining would detect infiltrating low-grade glioma cells that are not efficiently labeled by proliferative markers. Taken together, DCX is applicable for the detection of individual infiltrating glioma cells when combined with other markers. PMID- 18415659 TI - Radioimmunotherapy with [188Re]-labelled anti-CD66 antibody in the conditioning for allogeneic stem cell transplantation for high-risk acute myeloid leukemia. AB - Between July 2000 and June 2003 a total of 21 patients with high-risk acute myeloid leukemia (AML; n = 14), AML after myelodysplastic syndrome (MDS; n = 6) or advanced MDS (n = 1) were treated with an 188-Re labelled anti-CD66 antibody in the conditioning regimen for allogeneic stem cell transplantation. Radioimmunotherapy (RIT) was followed by standard full-dose conditioning with busulfan and high-dose cyclophosphamide in 11 patients and reduced intensity conditioning regimen in 10 patients. All patients received an unmanipulated allogeneic graft from alternative donors (n = 15) or a HLA-identical familiy donor (n = 6). With a median follow up of 42 months (23-60) disease free survival for all patients was 43%. Nine patients are still alive and in ongoing complete hematological remission. The treatment related mortality was 28.6% (n = 6) and an equal number of patients died of relapsing disease within 30-385 days after transplantation. Late organ toxicity, monitored for more than 1 year, was mild and not clinically relevant. The combination of RIT with chemotherapeutic conditioning seems to be a therapy with an acceptable risk of treatment related morbidity and mortality as well as occurrence of severe acute GvHD. PMID- 18415661 TI - Frequent and variable abnormalities in p14 tumor suppressor gene in glioma cell lines. AB - Ten glioma cell lines were examined for abnormalities of exon 1beta of the p14 gene and then for abnormalities of the entire p14 gene with the use of previous findings of other exons. Abnormalities of exon 1beta and the entire p14 gene were detected in eight of ten cases: homozygous deletion of the entire gene in six cases, hemizygous deletion of exon 1beta with homozygous deletion of downstream exons in one case, and hemizygous deletion of the entire coding region with a missense mutation (A97V) at the C-terminal nucleolar localization domain in one case. The remaining two cases revealed no such abnormalities. p14 gene expression was observed in the latter two cases and one case with A97V mutation in the hemizygously deleted coding region, but not in the others, including one case with only exon 1beta. In the three cases with p14 gene expression, immunocytochemistry revealed p14 nucleolar staining, suggesting the retention of the functional activity of p14 protein and, in the case with the A97V mutation, an insufficient mutational effect as well. The present findings of the frequent and variable p14 gene abnormalities, including rare-type ones with or without sufficient mutational effect in glioma cell lines, might be of value for better understanding of the p14 gene and its related pathways in glioma carcinogenesis. PMID- 18415662 TI - Ultrastructural study of neuronal and related tumors in the ventricles. AB - Intraventricular tumors may arise from a variety of cells in the region. There are some difficulties in diagnosing these tumors because of their histologically similar appearance. We analyzed intraventricular tumors, including central neurocytoma, oligodendroglioma, cerebral neuroblastoma, and cerebellar neuroblastoma, the neuronal characters of which were established based on their ultrastructural findings, except for oligodendroglioma. Central neurocytoma and cerebellar neuroblastoma showed synaptic formation, and cerebral neuroblastoma possessed immature neurites. Oligodendroglioma showed similar structures to that of a normal oligodendrocyte. Furthermore, we review the literature and evaluate the usefulness of analyzing ultrastructures. PMID- 18415663 TI - Consecutive histological changes in an astroblastoma that disseminated to the spinal cord after repeated intracranial recurrences: a case report. AB - A 17-year-old male patient underwent surgery five times (four consecutive intracranial tumor removal surgeries and a final spinal tumor removal surgery). After the third surgery, this case was reported as a low-grade astroblastoma that is characterized by perivascular pseudorosettes consisting of elongated tumor cells arranged around the blood vessels. However, the fourth and fifth surgical specimens demonstrated very interesting histological changes in the astroblastoma. Through the course of relapses, the constituent cells of the astroblastic perivascular rosettes became smaller and rounder, and a multilayered cell arrangement was observed. The nucleus-to-cytoplasm ratio increased, and the compact intervascular cells ultimately lost glial fibrillary acidic protein (GFAP) expression. These undifferentiated cells showed high MIB-1 indices and an increased olig2 index. On the other hand, the cells in all the surgical specimens were positive for certain neuronal markers such as NSE, TUJ1, and nestin. Some astroblastomas may be more immature than the usual astrocytes; however, it is necessary to study more cases to confirm this. PMID- 18415664 TI - A case of lymphomatoid granulomatosis followed for 14 months on the basis of clinical and histological findings. AB - Lymphomatoid granulomatosis (LYG) is a systemic granulomatous disease characterized by B-cell proliferation of uncertain malignant potential. It most frequently affects the lungs but also occasionally affects the central nervous system. Its pathophysiology is unclear in numerous respects, thus making it difficult to diagnose and treat. We recently encountered a case of LYG that was followed clinically and histologically for 14 months. A 55-year-old man was hospitalized with multiple brain tumors for which the final diagnosis was not made until the second surgery, 14 months after the first intervention. Following the diagnosis, he was treated with steroid pulse therapy. At present, 3.5 years after the onset of LYG, the patient is in good condition with no signs of tumor recurrence. Although LYG is usually graded on the basis of histological findings, this patient showed no histological changes or any increase in disease grade during the 14-month follow-up period. PMID- 18415665 TI - Medulloblastoma demonstrating multipotent differentiation: case report. AB - We report a 6-year-old boy who presented with a medulloblastoma demonstrating classic, myoblastic, neuronal, glial, and melanotic differentiation and manifesting as severe morning headache. Magnetic resonance imaging revealed a mass lesion with cystic components in the cerebellar vermis. He underwent suboccipital craniotomy and total resection of the tumor. The specimen consisted of three morphologically distinct components. The first component consisted of densely packed cells with round-to-oval highly hyperchromatic nuclei surrounded by scanty cytoplasm. Immunohistochemical staining revealed diffuse expression of neurofilament protein and focal expression of desmin and myoglobin. The second component consisted of long spindle-shaped cells with elongated nuclei and eosinophilic cytoplasm. Immunohistochemical staining revealed diffuse expression of neurofilament protein, desmin, and myoglobin. The third component consisted of cells with small, densely hyperchromatic nuclei and scanty cytoplasm in a fine fibrillary background. Mature ganglion cells and melanotic tumor cells were also observed. Immunohistochemical staining revealed diffuse expression of synaptophysin and neurofilament protein, and focal expression of glial fibrillary acidic protein, S-100 protein, desmin, and myoglobin. The diagnosis was medulloblastoma with myoblastic, neuronal, astrocytic, and melanotic differentiation. Medulloblastoma demonstrating multipotent differentiation is rare, but the features observed in this case support the idea that medulloblastoma originates from multipotent stem cells. PMID- 18415667 TI - The domains of psychiatric practice: from centre to periphery. PMID- 18415666 TI - A case of cerebral ganglioneuronal tumor in the parietal lobe of an adult. AB - Central nervous system (CNS) neuroblastoma/ganglioneuroblastoma is one of the embryonal tumors with neuronal differentiation found in young adults, but it is most common in children, especially in those below the age of 5 years, whereas extraventricular neurocytoma, a rare neuroepithelial tumor with neuronal differentiation, mostly affects young adults. Here we present a rare case of cerebral ganglioneuronal tumor that occurred in a 32-year-old woman. The patient suffered from tonic convulsion, and computed tomography demonstrated a well demarcated, round tumor 3.3 cm in size with marked calcification in the right parietal lobe. Histological analysis revealed diffuse infiltration of small, round cells with scattered large ganglion-like cells. Immunohistochemically, the tumor cells did not react with any neuronal molecules, except for chromogranin A in ganglion-like large tumor cells, but electron microscopy demonstrated the presence of synapse-like nerve terminal structures without mature postsynaptic density, suggesting the presence of neoplastic tumor components with neuronal differentiation; thus, this tumor was diagnosed as CNS ganglioneuroblastoma with possible low-grade malignancy because the Mib-1 labeling index was less than 3% 4%. Here we discuss the histological entity of cerebral ganglioneuronal tumors, including extraventricular neurocytoma. PMID- 18415668 TI - Titration of cardiolipin by either 10-N-nonyl acridine orange or acridine orange sensitizes the adenine nucleotide carrier to permeability transition. AB - Under the action of carboxyatractyloside or fatty acids, adenine nucleotide translocase switches its function from nucleotide carrier to modulator of the opening of a non-specific pore. In addition to the effect of these agents, this modification in activity is, in some way, dependent on the influence of the lipid milieu of the membrane. Cardiolipin is, among other membrane phospholipids, the one that interacts the most with the translocase. This work shows that 10-N-nonyl acridine orange and acridine orange, probes for this phospholipid, modify the sensitivity of the translocase to carboxyatractyloside, oleate, and palmitate to induce permeability transition. The results also show that these probes stimulate the release of mitochondrial cytochrome c, and increase labeling of the carrier by eosin 5-maleimide. Based on the aforementioned it is proposed that the increase in sensitivity is due to a conformational change in the translocase, induced by the binding of the probe to cardiolipin. PMID- 18415669 TI - Nicorandil protects cardiac mitochondria against permeability transition induced by ischemia-reperfusion. AB - Ischemia followed by reperfusion is known to negatively affect mitochondrial function by inducing a deleterious condition termed mitochondrial permeability transition. Mitochondrial permeability transition is triggered by oxidative stress, which occurs in mitochondria during ischemia-reperfusion as a result of lower antioxidant defenses and increased oxidant production. Permeability transition causes mitochondrial dysfunction and can ultimately lead to cell death. A drug able to minimize mitochondrial damage induced by ischemia reperfusion may prove to be clinically effective. We aimed to analyze the effects of nicorandil, an ATP-sensitive potassium channel agonist and vasodilator, on mitochondrial function of rat hearts and cardiac HL-1 cells submitted to ischemia reperfusion. Nicorandil decreased mitochondrial swelling and calcium uptake. It also decreased reactive oxygen species formation and thiobarbituric acid reactive substances levels, a lipid peroxidation biomarker. We thus confirm previous reports that nicorandil inhibits mitochondrial permeability transition and demonstrate that nicorandil inhibits this process by preventing oxidative damage and mitochondrial calcium overload induced by ischemia-reperfusion, resulting in improved cardiomyocyte viability. These results may explain the good clinical results obtained when using nicorandil in the treatment of ischemic heart disease. PMID- 18415670 TI - Influence of mitochondrial DNA level on cellular energy metabolism: implications for mitochondrial diseases. AB - The total amount of cellular mitochondrial DNA (mtDNA) varies widely and seems to be related to the nature and metabolic state of tissues and cells in culture. It is not known, however, whether this variation has any significance in vivo, and to which extent it regulates energy production. To better understand the importance of the cellular mtDNA level, we studied the influence of a gradual reduction of mtDNA copy number on oxidative phosphorylation in two models: (a) a control human cell line treated with different concentrations of 2', 3' dideoxycytidine, a nucleoside analogue that inhibits mtDNA replication by interfering with mitochondrial DNA polymerase gamma, and (b) a cell line derived from a patient presenting mtDNA depletion. The two models were used to construct biochemical and phenotypic threshold curves. Our results show that oxidative phosphorylation activities are under a tight control by the amount of mtDNA in the cell, and that the full complement of mtDNA molecules are necessary to maintain a normal energy production level. PMID- 18415671 TI - Prevalence of overweight and obesity among women of childbearing age: results from the 2002 National Survey of Family Growth. AB - OBJECTIVE: To estimate the prevalence of overweight and obesity among U.S. women of childbearing age. METHODS: Our study population was drawn from the 2002 National Survey of Family Growth (NSFG) and consisted of non-pregnant female respondents aged 20-44 years with a valid body mass index (BMI) (N = 5,958). Univariate and bivariate analyses were conducted to document variations in the prevalence of overweight and obesity by age, race and Hispanic origin, and socioeconomic status. RESULTS: Overall, 24.5% of women 20-44 years of age were overweight (BMI 25.0-29.9 kg/m(2)) and 23.0% were obese (BMI > or = 30.0 kg/m(2)). Among those who were obese, 10.3% met the criteria for class II or III obesity (BMI > or = 35.0 kg/m(2)). Non-Hispanic black and women were 2.25 times more likely to be overweight or obese compared to non-Hispanic white women (95% CI: 1.87-2.69). This disparity in risk between non-Hispanic black and non Hispanic white women declined and no longer achieved statistical significance after adjustment for education, household income, and health insurance coverage. CONCLUSIONS: Nearly one in two U.S. women of childbearing age are either overweight or obese. The racial/ethnic disparity in prevalence rates may be due in part to variability in educational attainment, household income, and stability of health insurance coverage. PMID- 18415672 TI - A new algorithm for concealed accessory pathway localization using T-wave subtracted retrograde P-wave polarity during orthodromic atrioventricular reentrant tachycardia. AB - INTRODUCTION: AP localization can be predicted by analyzing the polarity of the delta wave, QRS polarity, and R/S ratio in patients with Wolff-Parkinson-White syndrome. However, the estimation of AP location is limited in patients with concealed pathways during atrioventricular reentrant tachycardias (AVRT). Thus, we analyzed retrograde P-wave polarity during orthodromic AVRT and developed an algorithm to predict the localization of concealed accessory pathways (AP). METHODS AND RESULTS: A total number of 131 patients with a single AP and inducible orthodromic AVRT were included. The initial 61 patients were analyzed retrospectively for algorithm development, whereas 70 patients were evaluated prospectively. The retrograde P-wave polarity was analyzed by subtracting the superimposing T-wave during orthodromic AVRT using custom-designed software. Four leads of the surface electrocardiogram (ECG) were identified to accurately distinguish AP locations assigned to four different regions around each AV annulus: I, aVR, aVL, and V(1). Lead V(1) was used to differentiate right (negative or isoelectric) from left (solely positive) APs. Retrograde P-wave in lead I was negative in left posterior APs exclusively and became more positive with an AP location shifting towards right anterior. P-wave polarity in lead aVR demonstrated a shift from a positive polarity from left APs to isoelectric in right APs. The opposite direction (shift from positive to isoelectric) was observed for lead aVL. The subsequently developed algorithm for concealed AP localization using these surface ECG leads demonstrated a high sensitivity, specificity, and positive predictive value particularly for common AP localizations (left posterior and inferior, and right septal) when applied in a prospective fashion. CONCLUSION: Concealed AP localization can be accurately predicted by the analysis of retrograde P-wave polarity during orthodromic AVRT using the algorithm derived from the presented study. PMID- 18415673 TI - Wake up and smell the coffee: yet another no go for cardiac patients? : editorial to "caffeinated coffee blunts the myocardial protective effects of statins against ischemia-reperfusion injury in the rat" by Ye et al. PMID- 18415674 TI - DARPP-32 expression in rat brain after an inhibitory avoidance task. AB - Step-down inhibitory avoidance (IA) is usually acquired in one single trial, which makes it ideal for studying processes initiated by training, uncontaminated by prior or further trials, rehearsals, or retrievals. Biochemical events in the hippocampus related to long-term memory (LTM) formation have been extensively studied in rats using a one trial step-down IA task. DARPP-32 (dopamine and cAMP regulated phosphoprotein of Mr 32 kDa) is a cytosolic protein that is selectively enriched in medium spiny neurons in the neostriatum. It has been shown that activation of DARPP-32 and the resultant inhibition of PP-1 activity is critical for the expression of two opposing forms of brain synaptic plasticity, striatal LTD and LTP. Both forms of plasticity are also critically linked to the activation of DA receptors. It has been shown with studies in DARPP-32 KO mice an important role of this protein in mediating the effects of DA on long term changes in neuronal excitability and to our knowledge, no studies have examined the effect of IA task on DARPP-32 expression. In order to demonstrate changes in the protein expression profile we analyzed DARPP-32 levels in the striatum, prefrontal cortex (PFC), hippocampus and entorhinal cortex of Wistar rats after step-down IA learning. Our results showed that IA induced changes on DARPP-32 expression in striatum and hippocampus. DARPP-32 expression changes corroborate with changes in expression and phosphorylation of CREB, NMDA, AMPA after IA that has been reported. These changes suggest that DARPP-32 might play a central role in the IA, as previously described as an integrator of the dopaminergic signal. PMID- 18415675 TI - Modulation by clamping: Kv4 and KChIP interactions. AB - The rapidly inactivating (A-type) potassium channels regulate membrane excitability that defines the fundamental mechanism of neuronal functions such as pain signaling. Cytosolic Kv channel-interacting proteins KChIPs that belong to neuronal calcium sensor (NCS) family of calcium binding EF-hand proteins co assemble with Kv4 (Shal) alpha subunits to form a native complex that encodes major components of neuronal somatodendritic A-type K+ current, I(SA), in neurons and transient outward current, I(TO), in cardiac myocytes. The specific binding of auxiliary KChIPs to the Kv4 N-terminus results in modulation of gating properties, surface expression and subunit assembly of Kv4 channels. Here, I attempt to emphasize the interaction between KChIPs and Kv4 based on recent progress made in understanding the structure complex in which a single KChIP1 molecule laterally clamps two neighboring Kv4.3 N-termini in a 4:4 manner. Greater insights into molecular mechanism between KChIPs and Kv4 interaction may provide therapeutic potentials of designing compounds aimed at disrupting the protein-protein interaction for treatment of membrane excitability-related disorders. PMID- 18415678 TI - Colonic metastasis from hepatocellular carcinoma: manifested by gastrointestinal bleeding. PMID- 18415676 TI - Role of nitric oxide synthases in Parkinson's disease: a review on the antioxidant and anti-inflammatory activity of polyphenols. AB - Natural polyphenols can exert protective action on a number of pathological conditions including neurodegenerative disorders. The neuroprotective effects of many polyphenols rely on their ability to permeate brain barrier and here directly scavenge pathological concentration of reactive oxygen and nitrogen species and chelate transition metal ions. Importantly, polyphenols modulate neuroinflammation by inhibiting the expression of inflammatory genes and the level of intracellular antioxidants. Parkinson's disease (PD) is a neurodegenerative disorder characterized by several abnormalities including inflammation, mitochondrial dysfunction, iron accumulation and oxidative stress. There is considerable evidence showing that cellular oxidative damage occurring in PD might result also from the actions of altered production of nitric oxide (NO). Indeed, high levels of neuronal and inducible NO synthase (NOS) were found in substantia nigra of patients and animal models of PD. Here, we evaluate the involvement of NOS/NO in PD and explore the neuroprotective activity of natural polyphenol compounds in terms of anti-inflammatory and antioxidant action. PMID- 18415677 TI - Benefits from dietary polyphenols for brain aging and Alzheimer's disease. AB - Brain aging and the most diffused neurodegenerative diseases of the elderly are characterized by oxidative damage, redox metals homeostasis impairment and inflammation. Food polyphenols can counteract these alterations in vitro and are therefore suggested to have potential anti-aging and brain-protective activities, as also indicated by the results of some epidemiological studies. Despite the huge and increasing amount of the in vitro studies trying to unravel the mechanisms of action of dietary polyphenols, the research in this field is still incomplete, and questions about bioavailability, biotransformation, synergism with other dietary factors, mechanisms of the antioxidant activity, risks inherent to their possible pro-oxidant activities are still unanswered. Most of all, the capacity of the majority of these compounds to cross the blood-brain barrier and reach brain is still unknown. This commentary discusses recent data on these aspects, particularly focusing on effects of curcumin, resveratrol and catechins on Alzheimer's disease. PMID- 18415679 TI - Role of procalcitonin in infectious gastroenteritis and inflammatory bowel disease. AB - BACKGROUND AND AIM: We have evaluated procalcitonin (PCT) as a diagnostic marker for bacterial gastroenteritis (GE) and as a disease activity marker in inflammatory bowel disease (IBD) patients. METHODS: This was a prospective single center study performed over a 1-year period. Venous blood samples were drawn from hospitalized patients with acute GE and tested for PCT, C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), and total white cell count (TWC); stools from the same patients were tested for standard pathogens. Venous blood samples from patients with IBD were tested for PCT, CRP, ESR, and platelet count. The PCT level was measured using an immunofluorescent assay, with normal being defined as <0.5 ng/ml. RESULTS: The GE arm of study consisted of 81 patients, 18.5% of whom were diagnosed with bacterial GE. The PCT and CRP levels were good diagnostic markers of bacterial GE, with an area under the curve (AUC) of 0.727 [95% confidence interval (CI) 0.580-0.874] and 0.786 (95% CI 0.627-0.946), respectively. An elevated PCT > or =0.5 ng/ml was associated with a 13-fold increased risk of renal impairment. The IBD arm of study consisted of 72 IBD patients. The PCT levels were not significantly different between active and inactive IBD in this patient cohort. CONCLUSION: Our results indicate that PCT and CRP are comparably good diagnostic markers of bacterial GE but that PCT is not useful as in monitoring disease activity in patients with IBD. PMID- 18415680 TI - A novel diquinolonium displays preclinical anti-cancer activity and induces caspase-independent cell death. AB - Quinolines are a class of chemical compounds with emerging anti-cancer properties. Here, we tested the activity of series of quinolines and quinoline like molecules for anti-cancer activity and identified a novel diquinoline, 1 methyl-2-[3-(1-methyl-1,2-dihydroquinolin-2-yliden)prop-1-enyl]quinolinium iodide (Q(2)). Q(2 )induced cell death in leukemia, myeloma, and solid tumor cell lines with LD50s in the low to submicromolar range. Moreover, Q(2) induced cell death in primary acute myeloid leukemia (AML) cells preferentially over normal hematopoietic cells. In a mouse model of leukemia, Q(2) delayed tumor growth. Mechanistically, Q(2) induced cell death through caspase independent mechanisms. By electron microscopy, Q(2) increased cytoplasmic vacuolization and mitochondrial swelling. Potentially consistent with the induction of autophagic cell death, Q(2) treatment led to a punctate distribution of LC3 and increased MDC staining. Thus, Q(2) is a novel quinolinium with preclinical activity in malignancies such as leukemia and myeloma and warrants further investigation. PMID- 18415681 TI - Role of Bcl-2 family members in caspase-3/9-dependent apoptosis during Pseudomonas aeruginosa infection in U937 cells. AB - Pseudomonas aeruginosa is a gram-negative opportunistic pathogen that is cytotoxic towards a variety of eukaryotic cells. To investigate the effect of this bacterium on monocyte, we infected human U937 cells with the P. aeruginosa strain in vitro. To explore the expression of Bcl-2 and Bax as well as caspase 3/9 activation in the apoptosis of human U937 cells induced by P. aeruginosa, Hoechst 33258 staining and Giemsa staining as well as Flow cytometry analysis were used to determine the rate of apoptosis, and the expressions of Bcl-2 and Bax were assayed by RT-PCR and Western blotting respectively. Bax protein conformation change was assayed by immunoprecipitation. Cytochrome c release was measured by Western blotting. Moreover, exposure of U937 cells to P. aeruginosa measured caspase-3/9 activity. It was found that the apoptosis of human U937 cells could be induced by Pseudomonas aeruginosa in a dose- and time-dependent manner. Also, there were a tendency of alterations with an increased expression level of Bax and a reduced expression level of Bcl-2, increased levels of cytochrome c release, and also with an increased activation of caspase-3/9 and Bax protein conformation change. For the evaluation of the role of caspases, caspase-3/9 inhibitors Z-DEVD-FMK and Z-LEHD-FMK respectively were used. The results were further confirmed by the observation that the caspase inhibitors Z DEVD-FMK and Z-LEHD-FMK blocked P. aeruginosa-induced U937 apoptosis. It is concluded that P. aeruginosa can induce apoptosis with an up-regulated expression of Bax and a down-regulated expression of Bcl-2, which resulted in increased levels of cytochrome c release and increased caspase-3 and -9 in human U937 cells. PMID- 18415682 TI - A review of the stereotype threat literature and its application in a neurological population. AB - Stereotype threat is a situational phenomenon, leading to test performance decrements, in which a member of a stigmatized group feels pressured by the possibility of confirming or being judged by a negative stereotype. This review article highlights the progression of research in the stereotype threat field, and its relevance to neurological populations. Early studies focused on demonstrating this effect in African American, women, and elderly populations. Since this time, research has continued to focus on these populations but has moved to elucidating stereotype threat's mediating psychological factors, studying the impact of individual differences in response to stereotype threat, and attempting to reduce its overall effect. A proposal for further study in neurological populations, under the framework of stereotype threat, comprises the last portion of the paper. It is argued that this social psychological phenomenon may, at least in part, account for poor neuropsychological test performance for neurologically compromised individuals. PMID- 18415684 TI - Central hypothyroidism. AB - Central hypothyroidism (CH) is a rare cause of hypothyroidism due to an insufficient stimulation of an otherwise normal thyroid gland and it is caused by either pituitary (secondary hypothyroidism) or hypothalamic (tertiary hypothyroidism) defects. The diagnosis of CH is usually suggested by the finding of lowered thyroid hormone concentrations, associated with inappropriately low/ normal TSH levels. Restoration and maintenance of euthyroidism represent the therapeutic goals in all forms of CH. On these basis, the vast majority of patients with CH is treated with standard levo-thyroxine (L-T4) therapy which is tailored according to FT4 circulating levels that should be maintained in the normal range. PMID- 18415683 TI - Neuropsychological sequelae of non-central nervous system cancer and cancer therapy. AB - Cancer patients report numerous adverse symptoms associated with their disease and treatment including cognitive dysfunction, fatigue, and affective distress. Cognitive dysfunction is ubiquitous in patients with primary central nervous system (CNS) cancer and recent evidence has documented similar deficits in patients with non-CNS cancer as well. Both the cancer itself and treatments including chemotherapy, biological response modifiers, and hormonal therapies have been demonstrated to adversely impact cognitive and neurobehavioral function. Neuroimaging and neurophysiological investigations have likewise revealed alterations in brain function that are helping to account for the nature of these cognitive disorders. Similarly, preclinical animal research is assisting to identify the pathophysiological mechanisms that underlie treatment-related neurotoxicities. The coalescence of multidisciplinary clinical and research efforts hold promise for the development of interventions that may offer neuroprotection in addition to currently available symptomatic therapies and cognitive rehabilitation techniques. PMID- 18415685 TI - Data and public health decision making on HIV prevention in injection drug users. PMID- 18415686 TI - The low temperature-responsive, Solanum CBF1 genes maintain high identity in their upstream regions in a genomic environment undergoing gene duplications, deletions, and rearrangements. AB - Some plants like Arabidopsis thaliana increase in freezing tolerance when exposed to low nonfreezing temperatures, a process known as cold acclimation. Other plants including tomato, Solanum lycopersicum, are chilling sensitive and incur injury during prolonged low temperature exposure. A key initial event that occurs upon low temperature exposure is the induction of genes encoding the CBF transcription factors. In Arabidopsis three CBF genes, present in a tandemly linked cluster, are induced by low temperatures. Tomato also harbors three tandemly-linked CBF genes, Sl-CBF3-CBF1-CBF2, but only one of these, Sl-CBF1, is low-temperature responsive. Here we report that Solanum species that are closely allied to cultivated tomato essentially share this structural organization, but the locus is in a dynamic state of flux. Additional paralogs and in-frame deletions between adjacent genes occur, and the genomic regions flanking the CBF genes are dissimilar across Solanum species. Nevertheless, the CBF1 upstream region remains intact and highly conserved. This feature differed for CBF2 and CBF3, whose upstream regions were far less conserved. CBF1 was also the only low temperature responsive gene in the cluster and its expression was greatly affected by a circadian clock. The tuber-bearing S. tuberosum and S. commersonii also harbored a fourth gene, CBF4, which was also low temperature responsive. CBF4 was physically linked to CBF5 in S. tuberosum, but CBF5 was absent from S. commersonii. Phylogenic analyses suggest that CBF5-CBF4 resulted from the duplication of the CBF3-CBF1-CBF2 cluster. DNA sequence motifs shared between the Solanum CBF1 and CBF4 upstream regions were identified, portions of which were also present in the Arabidopsis CBF1-3 upstream regions. These results suggest that much greater functional constraints are placed upon the Solanum CBF1 upstream regions over the other CBF upstream regions and that CBF4 has retained the capacity for low temperature responsiveness following the duplication event that gave rise to CBF4. PMID- 18415687 TI - Radiofrequency electromagnetic fields; male infertility and sex ratio of offspring. AB - Concern is growing about exposure to electromagnetic fields and male reproductive health. The authors performed a cross-sectional study among military men employed in the Royal Norwegian Navy, including information about work close to equipment emitting radiofrequency electromagnetic fields, one-year infertility, children and sex of the offspring. Among 10,497 respondents, 22% had worked close to high frequency aerials to a "high" or "very high" degree. Infertility increased significantly along with increasing self-reported exposure to radiofrequency electromagnetic fields. In a logistic regression, odds ratio (OR) for infertility among those who had worked closer than 10 m from high-frequency aerials to a "very high" degree relative to those who reported no work near high-frequency aerials was 1.86 (95% confidence interval (CI): 1.46-2.37), adjusted for age, smoking habits, alcohol consumption and exposure to organic solvents, welding and lead. Similar adjusted OR for those exposed to a "high", "some" and "low" degree were 1.93 (95% CI: 1.55-2.40), 1.52 (95% CI: 1.25-1.84), and 1.39 (95% CI: 1.15 1.68), respectively. In all age groups there were significant linear trends with higher prevalence of involuntary childlessness with higher self-reported exposure to radiofrequency fields. However, the degree of exposure to radiofrequency radiation and the number of children were not associated. For self-reported exposure both to high-frequency aerials and communication equipment there were significant linear trends with lower ratio of boys to girls at birth when the father reported a higher degree of radiofrequency electromagnetic exposure. PMID- 18415688 TI - The spatial and temporal distribution of predatory and phytophagous mites in field-grown strawberry in the UK. AB - Extensive sampling of strawberry plants in everbearing and June-bearing strawberry plantations and on potted plants showed that different species of mites were spatially separated. Of the two phytophagous species recorded, Tetranychus urticae was most abundant on old leaves and Phytonemus pallidus on folded leaves and flower/fruit clusters. Predatory phytoseiid mites were found on all plant parts but different species were spatially separated; Neoseiulus cucumeris and N. aurescens were found mostly on folded leaves and clusters, and N. californicus and Phytoseiulus persimilis on old and medium aged leaves. No Typhlodromus pyri were found in the field plantations. These patterns of distribution did not change over sampling dates in summer and early autumn. An understanding of this within-plant zonation of mite species is important when studying predator-prey interactions and when designing sampling strategies for strawberry. A programme to sample the entire mite system on strawberry should be stratified to include all the above mentioned parts of the plant. Different sampling protocols, as appropriate, are required for sampling different pest species and their associated predators. PMID- 18415689 TI - Developmental patterns of caspase-3, bax and bcl-2 proteins expression in the human spinal ganglia. AB - The distribution of the bcl-2, bax and caspase-3 proteins was investigated in the cells of developing human spinal ganglia. Paraffin sections of 10 human conceptuses between 5th and 9th gestational weeks were analysed morphologically, immunohistochemically and by TUNEL-method. Cells positive to caspase-3 had brown stained nuclei or nuclear fragmentations. At earliest stages, 6% of ganglion population were caspase-3 positive cells. Later on, a significant increase in number of caspase-3 positive cells appeared, particularly in the ventral part of ganglia (12%), and subsequently decreased to 6%. TUNEL-positive cells had the same distribution pattern as caspase-3 positive cells. Bax-positive cells followed the developmental pattern similar to caspase-3 cells, changing in range between 20% and 32%. There were 8% of bcl-2 positive cells at earliest stages. They increased significantly in dorsal part of the ganglion during the 7th week (28%), and than dropped to 15% by the end of the 8th week. These findings suggest a ventro-dorsal course of development in human spinal ganglia. Number of bcl-2, bax and caspase-3 positive cells changed in a temporally and spatially restricted manner, coincidently with ganglion differentiation. While apoptosis might control cell number, bcl-2 could act in suppression of apoptosis and enhancement of cell differentiation. PMID- 18415690 TI - Genetic variation in hormone metabolizing genes and risk of testicular germ cell tumors. AB - Testicular germ cell tumors (TGCT) that arise in young men are composed of two histologic types, seminomas and nonseminomas. Risk patterns for the two types appear to be similar and may be related to either endogenous or exogenous hormonal exposures in utero. Why similar risk patterns would result in different histologic types is unclear, but could be related to varying genetic susceptibility profiles. Genetic variation in hormone metabolizing genes could potentially modify hormonal exposures, and thereby affect which histologic type a man develops. To examine this hypothesis, 33 single nucleotide polymorphisms (SNPs) in four hormone metabolism candidate genes (CYP1A1, CYP17A1, HSD17B1, HSD17B4) and the androgen receptor gene (AR) were genotyped. Associations with TGCT were evaluated among 577 TGCT cases (254 seminoma, 323 nonseminoma) and 707 controls from the US Servicemen's Testicular Tumor Environmental and Endocrine Determinants (STEED) study. There were no significant associations with TGCT overall based on a test using an additive model. However, compared to homozygotes of the most common allele, two nonredundant SNPs in CYP1A1 were inversely associated with nonseminoma: CYP1A1 promoter SNP rs4886605 OR = 0.75 (95% CI = 0.54-1.04) among the heterozygotes and OR = 0.37, 95% CI = 0.12-1.11 among the homozygotes with a p-value for trend = 0.02; rs2606345 intron 1 SNP, OR = 0.69 (95% CI = 0.51-0.93) among heterozygotes and OR = 0.70 (95% CI = 0.42-1.17) among homozygotes, with a p-value for trend = 0.02. Caution in interpretation is warranted until findings are replicated in other studies; however, the results suggest that genetic variation in CYP1A1 may be associated with nonseminoma. PMID- 18415691 TI - Akt1 is necessary for the vascular maturation and angiogenesis during cutaneous wound healing. AB - Previous in vivo and in vitro studies have shown that Akt1 serves as a crucial regulator of vascular maturation, extracellular matrix composition, and angiogenesis in tumors. Hence, we hypothesized that Akt1 may be necessary for other angiogenesis-dependent processes, including wound healing. Using Akt1 (-/-) and Akt2 (-/-) mice, we demonstrate that deficiency of Akt1, but not Akt2, results in impaired assembly of collagen in skin wounds and around the blood vessels. Wounds in Akt1 (-/-) mice, but not in Akt2 (-/-) mice, were characterized by reduced vascular area as well as impaired vascular maturation as evidenced by reduced smooth muscle cell recruitment. Expression level of a major angiogenic growth factor, VEGF, was significantly lower in wound tissues of Akt1 (-/-) mice as compared to WT. However, despite the impaired collagen assembly and reduced angiogenesis in Akt1 (-/-) wounds, no significant difference in migration of fibroblasts into the wound area was observed between WT and Akt1 (-/-) mice. Importantly, the dynamics of wound closure were similar between WT, Akt1 (-/-), and Akt2 (-/-) mice. Thus, it appears that although the lack of Akt1 impairs VEGF expression, wound angiogenesis, and subsequent maturation of vasculature, it has no effect on the wound closure. These findings may have clinical applications for the improvement of treatment procedures with reported history of wound healing complications. PMID- 18415692 TI - Analysis of insecticides in okra and brinjal from IPM and non-IPM fields. AB - Samples of okra and brinjal fruits, collected from non-integrated pest management (Non-IPM) and IPM fields in village Raispur, Ghaziabad District (U.P.), were analyzed for pesticide residues. The residues of chlorpyrifos in soil were 4.219 and 1.135 microg/g at harvest time in non-IPM and IPM fields of summer okra crop from initial value of 0.407 microg/g before sowing, while in brinjal crop, it was not detected in soils of any trials. During first year of study, the residues of chlorpyrifos and cypermethrin in okra fruit were observed to be 5.75 and 0.625 microg/g, respectively, for non-IPM fields; and 0.104 microg/g of chlorpyrifos for IPM trials. The pesticide residues were found to be 0.77, 1.39, 0.4 and 0.32 microg/g for cypermethrin, chlorpyrifos, monocrotophos and dimethoate, respectively, for non-IPM okra fruits in second year. For brinjal fruit, residues of cypermethrin and imidacloprid were not detected in IPM trials while it was found to be 0.28 and 0.78 microg/g for cypermethrin and chlorpyrifos respectively, for non-IPM trials. PMID- 18415693 TI - Assessing the performance of wastewater treatment with the combination of Fenton and ferrite process. AB - Industrial wastewater typically contains various metal ions. Traditional metal ion treatment processes such as chemical precipitation generate large volumes of toxic sludge which needs to be further solidified or disposed of. The ferrite process (FP), which is another effective approach of treating metal ion containing wastewater, can crystallize metal ions into ferrites; the sludge easily precipitates, is stable and can be recycling. This investigation explores the feasibility of the Fenton process and the FP (FFP) for treating wastewater that contains metal ions. It considers one factor that adds noise to the FP, ethylenediaminetetraacetic acid, and establishes the optimum parameters of each procedure. The analytical results demonstrate that the proper conditions for Fenton process were pH = 2, [Fe(2+)] = 10(-2) M, H(2)O(2) dosing rate = 5 x 10( 4) mol min(-1), reaction time = 12 min. For the proposed multi-stage FP, the preferred Fe(2+) dosage was 0.098 mol in each stage and the sludge met the toxicity characteristic leaching procedure standards. Following the FFP, the effluent water or the sludge easily met Taiwan's standards. Finally, the SEM test demonstrated that size of the sludge particles was 50-110 nm. PMID- 18415694 TI - The impact of the tunnel exhausts in terms of heavy metals to the surrounding ecosystem. AB - Samples of soil, plants, and lichens were analysed for heavy metal content (Cd, Cr, Cu, Ni, Pb, and Zn) in relation to different distances from the tunnel ventilation systems in order to evaluate the environmental pollution caused by car exhausts pollution from the Tauerntunnel and the Katschbergtunnel (Austria). Results show that the extent of heavy metal pollution is related to the type of tunnel ventilation system. The vertical ventilation system which ends in an exhaust air tower in the alpine pasture of Mosermandl 1,900 m above sea level contaminates soils and plants up to a distance of 750 m from the exhaust source. The dispersion of metals in front of the horizontal ventilation systems, which are located next to the tunnel portals, exhibits a rapid decrease with distance. Lichen transplants placed in front of the horizontal ventilation systems show very high metal accumulation, which gradually declines with the distance from the emitter source, therefore the lichen species Pseudevernia furfuracea can be used as a very efficient biomonitor for monitoring heavy metal pollution caused by tunnel exhausts over time. Data from former years (1986, 1988, 1990 and 1992) and from this present research suggest that the contamination of heavy metals at Mosermandl has decreased slightly in soil samples, except for Cu, but increased in plant samples, whereas a significant increase of Pb was observed. PMID- 18415695 TI - Evaluation of frequently used drug interaction screening programs. AB - OBJECTIVE: Drug-drug interaction (DDI) screening programs are an important tool to check prescriptions of multiple drugs. The objective of the current study was to critically appraise several DDI screening programs. METHODS: A DDI screening program had to fulfil minimal requirements (information on effect, severity rating, clinical management, mechanism and literature) to be included into the final evaluation. The 100 most frequently used drugs in the State Hospital of Baden, Switzerland, were used to test the comprehensiveness of the programs. Qualitative criteria were used for the assessment of the DDI monographs. In a precision analysis, 30 drugs with and 30 drugs without DDIs of clinical importance were tested. In addition, 16 medical patient profiles were checked for DDIs, using Stockley's Drug Interactions as a reference. MAIN OUTCOME MEASURE: Suitability of DDI screening program (quality of monographs, comprehensiveness of drug list, statistical evaluation). RESULTS: Out of nine programs included, the following four fulfilled the above mentioned criteria: Drug Interaction Facts, Drug-Reax, Lexi-Interact and Pharmavista. Drug Interaction Facts contained the smallest number of drugs and was therefore the least qualified program. Lexi Interact condenses many DDIs into one group, resulting in less specific information. Pharmavista and Drug-Reax offer excellent DDI monographs. In the precision analysis, Lexi-Interact showed the best sensitivity (1.00), followed by Drug-Reax and Pharmavista (0.83 each) and Drug Interaction Facts (0.63). The analysis of patient profiles revealed that out of 157 DDIs found by all programs, only 18 (11%) were detected by all of them. No program found more than 50% of the total number of DDIs. A further evaluation using Stockley's Drug interactions as the gold standard revealed that Pharmavista achieved a sensitivity of 0.86 (vs Drug Interaction Facts, Lexi-Interact and Drug-Reax with a sensitivity of 0.71 each) and a positive predictive value of 0.67. CONCLUSION: None of the four DDI screening programs tested is ideal, every program has its strengths and weaknesses, which are important to know. Pharmavista offers the highest sensitivity of the programs evaluated with a specificity and positive predictive value in an acceptable range. PMID- 18415696 TI - Fitting light saturation curves measured using modulated fluorometry. AB - A blue diode PAM (Pulse Amplitude Modulation) fluorometer was used to measure rapid Photosynthesis (P) versus Irradiance (E) curves (P vs. E curves) in Synechococcus (classical cyanobacteria), Prochlorothrix (prochlorophyta), Chlorella (chlorophyta), Rhodomonas (cryptophyta), Phaeodactylum (bacillariophyta) Acaryochloris (Chl d/a cyanobacteria) and Subterranean Clover (Trifolium subterraneum, Papilionaceae, Angiospermae). Effective quantum yield (Phi(PSII)) versus irradiance curves could be described by a simple exponential decay function (Phi(PSII) = Phi(PSII, maxe(-kE)) although Log/Log transformation was sometimes found to be necessary to obtain the best fits. Photosynthesis was measured as relative Electron Transport Rate (rETR) standardised on a chlorophyll basis. P versus E curves were fitted to the waiting-in-line function (an equation of the form P = P(max) x k x E x e(-kE)) allowing half-saturating and optimal irradiances (E(optimum)) to be estimated. The second differential of the equation shows that at twice optimal light intensities, there is a point of inflection in the P versus E curve. Photosynthesis is inhibited 26.4% at this point of inflection. The waiting-in-line model was found to be a very good descriptor of photosynthetic light saturation curves and superior to hyperbolic functions with an asymptotic saturation point (Michaelis-Menten, exponential saturation and hyperbolic tangent). The exponential constants (k) of the Phi(PSII) versus E and P versus E curves should be equal because rETR is directly proportional to Phi(PSII) x E. The conventionally calculated Non-Photochemical Quenching (NPQ) in Synechococcus was not significantly different to zero but NPQ versus E curves for the other algae could be fitted to an exponential saturation model. The kinetics of NPQ does not appear to be related to the kinetics of Phi(PSII) or rETR. PMID- 18415697 TI - Apoprotein A-V: an important regulator of triglyceride metabolism. AB - Apolipoprotein A-V (apoA-V) was discovered in 2001 both by comparative sequencing and as a liver regeneration protein. The gene is a located at the APOA1/C3/A4/A5 gene cluster on chromosome 11q23, a locus well known for playing a major role in regulating plasma cholesterol and triglyceride (TG) levels. ApoA-V is produced in the liver and has very low plasma concentrations (0.1-0.4 mug/ml). Mice lacking apoA-V have 4-fold increased TG levels, whereas apoA-V overexpression leads to 40% plasma TG reduction. Based on metabolic studies in vivo, apoA-V enhances the catabolism of TG rich lipoproteins rather than affecting their intestinal or hepatic production. By activating proteoglycans-bound lipoprotein lipase (LPL), apoA-V can accelerate TG hydrolysis from VLDL and chylomicrons independent from other apoproteins. Several variants at the APOA5 gene locus have been detected in humans. Some single nucleotide polymorphisms (SNPs) are associated with significantly higher plasma TG levels in patients (e.g., -1131T > C, S19W, G185C). In addition, these SNPs may affect fibrate response and obesity. However, data for a possible association of APOA5 variants with coronary heart disease are not consistent. Severe structural mutations (Q139X, Q148X, IVS3 + 3G > C) predispose to familial hypertriglyceridaemia and late-onset chylomicronaemia. Thus, despite its low plasma concentration, apoA-V is a major regulator of plasma TG metabolism in humans. However, the precise mechanism of its function is not yet clear. PMID- 18415698 TI - Inherited epithelial transporter disorders--an overview. AB - In the late 1990s, the identification of transporters and transporter-associated genes progressed substantially due to the development of new cloning approaches such as expression cloning and, subsequently, to the implementation of the human genome project. Since then, the role of many transporter genes in human diseases has been elucidated. In this overview, we focus on inherited disorders of epithelial transporters. In particular, we review genetic defects of the genes encoding glucose transporters (SLC2 and SLC5 families) and amino acid transporters (SLC1, SLC3, SLC6 and SLC7 families). PMID- 18415699 TI - Cerebral MRI as a valuable diagnostic tool in Zellweger spectrum patients. AB - Patients with defects in the biogenesis of peroxisomes include those with Zellweger syndrome spectrum (ZSS), a developmental and progressive metabolic disease with a distinct dysmorphic phenotype and varying severity. The diagnosis of ZSS relies on the clinical presentation and the biochemical evaluation of peroxisomal metabolites. Mutation detection in one out of twelve genes coding for proteins involved in the biogenesis of peroxisomes confirms the diagnosis. In the absence of pronounced clinical features of ZSS, neuroradiological findings may lead the way to the diagnosis. Cerebral magnetic resonance imaging (cMRI) pathology in ZSS consists of abnormal gyration pattern including polymicrogyria and pachygyria, leukencephalopathy, germinolytic cysts and heterotopias as reported by previous systematic studies including cMRI of a total of 34 ZSS patients, only five of whom had a severe phenotype. The present study evaluated the cMRI results of additional 18 patients, 6 with a severe and 12 with a milder ZSS phenotype. It confirms and extends knowledge of the characteristic cMRI pattern in ZSS patients. Besides an abnormal gyration pattern and delayed myelination or leukencephalopathy, brain atrophy was a common finding. Polymicrogyria and pachygyria were more common in patients with severe ZSS, while leukencephalopathy increases with age in patients with longer survival. Nevertheless, an abnormal gyration pattern might be more frequent in patients with a mild ZSS than deduced from previous studies. In addition, we discuss the differential diagnosis of the ZSS cMRI pattern and review investigations on the pathogenesis of the ZSS cerebral phenotype in mouse models of the disease. PMID- 18415700 TI - L-2-hydroxyglutaric aciduria: identification of ten novel mutations in the L2HGDH gene. AB - L-2-hydroxyglutaric aciduria (L-2-HGA) is a metabolic disease with an autosomal recessive mode of inheritance. It was first reported in 1980. Patients with this disease have mutations in both alleles of the L2HDGH gene. The clinical presentation of individuals with L-2-HGA is somewhat variable, but affected individuals typically suffer from progressive neurodegeneration. Analysis of urinary organic acids reveals an increased signal of 2-hydroxyglutaric acid, mainly as the L-enantiomer. L-2-HGA is known to occur in individuals of various ethnic backgrounds, but up to now mutation analysis has been mainly focused on patients of Turkish and Portuguese origin. This led us to confirm the diagnosis on the DNA level and undertake the corresponding mutation analysis in individuals of diverse ethnicity previously diagnosed with L-2-HGA on the basis of urinary metabolites and clinical/neuroimaging data. In 24 individuals from 17 families with diverse ethnic and geographic origins, 13 different mutations were found, 10 of which have not been reported previously. At least eight of the patients were compound heterozygotes. The identification of two mutations (c.751C > T and c.905C > T in exon 7) in patients with different origins supports the view that they occurred independently in different families. In contrast, the mutation c.788C > T was detected in all six Venezuelan patients originating from the same Caribbean island of Margarita, but not in other patients, thus rendering a founder effect likely. None of the mutations was found in the control population, indicating that they are most probably causative. Mutation analysis may improve the quality of diagnosis and prenatal diagnosis of L-2-HGA. PMID- 18415701 TI - Reduced carbohydrate intake in citrin-deficient subjects. AB - Citrin is the liver-type aspartate-glutamate carrier that resides within the inner mitochondrial membrane. Citrin deficiency (due to homozygous or compound heterozygous mutations in the gene SLC25A13) causes both adult-onset type II citrullinaemia (CTLN2) and neonatal intrahepatic cholestasis (NICCD). Clinically, CTLN2 is characterized by hyperammonaemia and citrullinaemia, whereas NICCD has a much more varied and transient presentation that can include multiple aminoacidaemias, hypoproteinaemia, galactosaemia, hypoglycaemia, and jaundice. Personal histories from CTLN2 patients have repeatedly described an aversion to carbohydrate-rich foods, and clinical observations of dietary and therapeutic outcomes have suggested that their unusual food preferences may be directly related to their pathophysiology. In the present study, we monitored the food intake of 18 Japanese citrin-deficient subjects whose ages ranged from 1 to 33 years, comparing them against published values for the general Japanese population. Our survey confirmed a marked decrease in carbohydrate intake, which accounts for a smaller proportion of carbohydrates contributing to the total energy intake (PFC ratio) as well as a shift towards a lower centile distribution for carbohydrate intake relative to age- and sex-matched controls. These results strongly support an avoidance of carbohydrate-rich foods by citrin-deficient patients that may lead to worsening of symptoms. PMID- 18415702 TI - Mefolinate (5-methyltetrahydrofolate), but not folic acid, decreases mortality in an animal model of severe methylenetetrahydrofolate reductase deficiency. AB - Severe deficiency of methylenetetrahydrofolate reductase (MTHFR) results in homocystinuria, with a variety of neurological and vascular complications, and sometimes death in the first year of life. MTHFR (EC 1.5.1.20) catalyses the synthesis of 5-methyltetrahydrofolate (5-methylTHF) which is required for homocysteine remethylation to methionine. Mthfr (-/-) mice are a good animal model of severe MTHFR deficiency in humans. They have marked hyperhomocysteinaemia and a high rate of mortality in the neonatal period. We attempted to rescue Mthfr (-/-) mice from postnatal death by treating their Mthfr (+/-) mothers with mefolinate (a synthetic form of 5-methylTHF, dissolved in their drinking water) or with a folic acid-enriched diet throughout pregnancy and lactation. We monitored pups' vitality and body weights until 3 weeks of age. The majority of Mthfr (-/-) pups from the control groups died during the first week of life. Body weights of -/- pups from control groups were significantly less than those of their Mthfr (+/-) and Mthfr ( +/+ ) littermates. Mefolinate treatment significantly improved survival rates (64% survival) in the -/- pups and improved morphology of the cerebellum. Folic acid supplementation did not affect the survival rate or body weights of the -/- pups. Our study suggests that MTHFR is important for postnatal growth and vitality, and that 5-methylTHF deficiency contributes to the high postnatal mortality. Mefolinate may be a good candidate drug for treatment of severe MTHFR deficiency. PMID- 18415703 TI - Predictors for aneurismal formation. PMID- 18415704 TI - Molecular identification and phylogenetic relationships of seven Indian Sciaenids (Pisces: Perciformes, Sciaenidae) based on 16S rRNA and cytochrome c oxidase subunit I mitochondrial genes. AB - The partial sequences of 16S rRNA and cytochrome c oxidase subunit I (COI) mitochondrial genes were analyzed for species identification and phylogenetic relationships among the commercially important Indian sciaenids (Otolithes cuvieri, Otolithes ruber, Johnius dussumieri, Johnius elongatus, Johnieops vogleri, Otolithoides biauritus and Protonibea diacanthus). Sequence analysis of both genes revealed that the seven species fell into three distinct groups, which were genetically distant from each other and exhibited identical phylogenetic resolution. Partial sequences of both the genes provided sufficient phylogenetic information to distinguish the seven sciaenids indicating the usefulness of mtDNA based approach in species identification. PMID- 18415705 TI - Rasch analysis of the short form 8-item Parkinson's Disease Questionnaire (PDQ 8). AB - PURPOSE: To evaluate the Italian version of the 8-item Parkinson's Disease Questionnaire (PDQ-8)--a subset of PDQ-39 (a 39-item health-related quality of life instrument for subjects with Parkinson's Disease [PD])--through classical psychometric techniques and Rasch analysis. METHODS: Two convenience samples (100 PD subjects each) were observed consecutively from 2004 to 2006. One group completed the PDQ-8 nested within PDQ-39, the other, the stand-alone PDQ-8. RESULTS: Once verified that the two independent samples came from the same population and showed consistent item calibrations using differential item functioning analysis, the two groups were combined. Cronbach's alpha was 0.72. According to Rasch analysis, the response scale of PDQ-8 could be simplified into a 3-category rating scale. After that, all the PDQ-8 items fitted the construct that the scale was intended to measure. Item separation reliability of PDQ-8 was 0.98 and person separation reliability was 0.70. Principal component analysis on the standardized residuals suggested a minor departure in the data from Rasch criteria (multidimensionality) and some marginal inter-item dependency. CONCLUSIONS: The PDQ-8 embedded in the PDQ-39 presented psychometric properties similar to the stand-alone PDQ-8. Our results, while consistent with previous classical psychometric analyses, add information on the meaningfulness of PDQ-8 in people with PD. In particular, a simplification of its rating scale is recommended. Moreover, additional analyses should be performed in order to further check unidimensionality and local dependence, and try to improve item selection and scaling properties of the questionnaire. In order to use the PDQ-8 for clinical decision-making in reference to individuals, its reliability should first be increased. PMID- 18415706 TI - Validation of Chinese Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) in patients scheduled for total knee replacement. AB - OBJECTIVE: Our aim was to cross-culturally validate Chinese Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) in patients with knee osteoarthritis (OA) scheduled for total knee replacement in Singapore. METHODS: Chinese WOMAC was translated from the original English version following standard guidelines. Patients were asked to complete a questionnaire containing the WOMAC (twice within 6 days), the Short Form 36 (SF-36), and the EuroQoL EQ-5D. Reliability was assessed using Cronbach's alpha and intraclass correlation coefficients (ICC), dimensionality using item-to-domain correlations and factor analyses. Convergent and discriminant construct validity was assessed using six each a priori hypotheses. RESULTS: Chinese WOMAC was well accepted by the patients in the pilot test. The Chinese and English versions were therefore administered to a consecutive sample of 131 Chinese- and 127 English-speaking subjects, respectively, with knee OA. Cronbach's alpha exceeded 0.7 for all domains except for Chinese pain domain, whereas the ICC exceeded 0.7 for all domains. Hypothesized item-to-domain correlations were observed for all items except for four items in Chinese physical function domain. The factor analyses yielded seven and five factors with eigenvalues of more than 1.0 in the Chinese and English versions, respectively. A total of 10/12 a priori construct validity hypotheses were satisfied for the Chinese version. Weak correlations between WOMAC pain and SF-36 bodily pain and moderate correlation between WOMAC pain and EQ-5D anxiety/depression were observed. CONCLUSIONS: Chinese WOMAC was well accepted and demonstrated acceptable psychometric properties in Singaporean patients with severe knee OA. PMID- 18415707 TI - Care recipients' perceptions of activity-related life space and life satisfaction during and after geriatric rehabilitation. AB - The debate concerning older people's life spaces should be based on subjective priorities of the elderly themselves. The purpose of this study was therefore to improve the understanding of preferences of elderly care recipients regarding activity-related life space (ARLS) and life satisfaction. A mainly qualitative design was used. Fifteen persons aged 80-94 years, undergoing geriatric rehabilitation, were interviewed during hospital stay and on two follow-up occasions after discharge. Transcribed interviews were analyzed in line with the thematic framework approach. The results point to three approaches related to preferences of ARLS: hierarchical limitations, changing continuity, and boundary breaking. Adaptive approaches were employed when physical incapacity was considered a hindrance to activity, adaptations which as a rule resulted in limitations of ARLS preferences. Activity related to the area 'close to one's own body' emerged as one of three identified key activities with importance for life satisfaction, the others being socializing and going out of doors. Continuity of activity in a familiar life space was expressed as a common ideal. If the aim of geriatric rehabilitation is to improve care recipients' life satisfaction, attention needs to be paid to the subjective dimensions of the ARLS in the goal setting. PMID- 18415708 TI - Differential cytokine pattern in the exhaled breath of patients with lung cancer. AB - Tumour cells may alter the protein pattern of biological samples resulting in specific differences that may aid diagnosis and treatment. In this pilot study we tested the cytokine pattern of exhaled breath condensate of patients with lung cancer. Breath condensates collected from 50 smoking patients with lung cancer and 25 smokers without clinical or radiological sign of a pulmonary tumour but having co-morbidities with similar severity as those with lung cancer were pooled for antibody microarray analysis testing 120 cytokines in parallel. Every cytokine on the array gave a signal in both groups. Nine cytokines including eotaxin, FGFs, IL-10 and MIP-3 were present with more than two-fold difference between the two groups. Large number of cytokines is present in the exhaled breath. Further analysis of specific differences associated with lung cancer may have clinical importance. PMID- 18415709 TI - Expression of p21(waf1/cip1), p27 (kip1), p63 and androgen receptor in low and high Gleason score prostate cancer. AB - The aim of this study was to investigate the expression of p21(waf1/cip1), p27(kip1), p63 and androgen receptor proteins in relation to serum prostate specific antigen levels in low and high Gleason score prostate cancers. Biopsies of patients suffering from prostate adenocarcinoma of low (3 + 3 to 3 + 4) and high (5 + 4 to 5 + 5) Gleason scores (13 cases each group) were immunostained for positive regulators of cell cycle control (p21(waf1/cip1) and p27(kip1)), and essential markers of normal prostate gland ontogeny (p63) and growth (androgen receptor) to find differentially expressed markers of malignant progression. Serum prostate specific antigen levels were also monitored at the time of biopsy and following anti-androgen therapy. All cases except one in each group were androgen receptor positive. P63 and p21(waf1/cip1) proteins detected in normal basal cell nuclei were lost in all but one studied tumors respectively. P27(kip1) protein, however, was detected in all low Gleason score prostate cancers, but it was found in only 7/13 high score cases. Prostate specific antigen levels, either pre- or post-treatment, did not show strict correlation with the p27(kip1) results. The low to high grade dedifferentiation of prostate adenocarcinoma is accompanied with the down-regulation of p27(kip1) protein, which may be an important molecular sign of the lost cell cycle control. PMID- 18415710 TI - The significance of Galectin-3 expression in the immunocytochemical evaluation of thyroid fine needle aspiration cytology. AB - The aim of this study is to evaluate the significance of immunohistochemical expression of Galectin-3 in the differential diagnosis of benign and malignant thyroid nodules. We studied the fine needle aspiration specimens of 38 patients who had evaluated for nodular goiter and undergone a thyroid surgery between 2004 2005. Slides had been stained immunocytochemically with Galectin-3. The cytoplasmic staining of Galectin-3 was analyzed. Three cases of five follicular carcinomas had positive staining for Galectin-3, while two had not. Two cases with follicular adenomas were negative for Galectin-3. Five cases of six papillary carcinomas had positive staining for Galectin-3, while one case (the case with a papillary microcarcinoma) had not. The single cases with medullary and anaplastic carcinomas were negative for Galectin-3. None of the cases with a benign thyroid pathology had positive staining for Galectin-3. Galectin-3 immunocytochemical staining, had a sensitivity of 61.5%, specificity of 100%, positive predictive value of 100%, and negative predictive value of 83.3% for thyroid malignancies. For the evaluation of follicular neoplasm, Galectin-3 immunocytochemical staining had a sensitivity of 60%, specificity of 100%, positive predictive value of 100%, and negative predictive value of 50%. Galectin 3 expression in thyrocytes is a strong indicator of a malignant proliferative lesion especially for papillary and to an extent in follicular thyroid neoplasms. Galectin-3 could be used as a supplementary marker for cytological diagnosis. PMID- 18415711 TI - Peritumoral retraction clefting correlates with advanced stage squamous cell carcinoma of the esophagus. AB - The present study was designated to analyze correlation between the presence and extent of peritumoral retraction clefting and various clinicopathologic features in esophageal squamous cell carcinoma (ESCC), and to possibly establish the significance of this phenomenon in ESCC. Fifty-four consecutive patients with advanced ESCC were included in the study. The presence of peritumoral retraction clefting was classified on the basis of the proportion of tumor nests exhibiting this phenomenon. Tumors with clefts that affected up to 25% of tumor nests were classified as group I; with clefts that affected >25% to 50% of tumor nests as group II; with clefts that affected >50% to 75% of tumor nests as group III; and tumors with clefts that affected more than 75% of tumor nests were classified as group IV. Statistical analysis showed a correlation between presence and extent of peritumoral clefting and lymph node metastasis. T3 tumors and tumors with lymph node metastasis had significantly more pronounced peritumoral clefting compared with T2 tumors and tumors without lymph node metastasis. The presence of peritumoral clefting was not associated with the number of affected lymph nodes. There was no correlation between the presence and extent of peritumoral clefting with patient age and sex, and tumor location, diameter and grade. The association of peritumoral retraction clefting in ESCC with local invasiveness and lymph node metastasis indicated that peritumoral clefting could be a simple and useful morphological feature of tumor aggressiveness and may contribute to the pathological and clinical assessment of patients with ESCC. PMID- 18415712 TI - Polymorphisms of the XRCC1 and XPD genes and breast cancer risk: a case-control study. AB - The purpose of this case control study was to evaluate the role of X-ray repair cross-complementing group 1 (XRCC1) and xeroderma pigmentosum group D (XPD) genotypes as genetic indicators of susceptibility to breast cancer (BC). We analysed DNA samples from 114 breast cancer patients and 113 control subjects using polymerase chain reaction-restriction fragment length polymorphism. For the single nucleotide polymorphisms in XRCC1 exon 10 (Arg399Gln, G/A) and XPD exon 23 (Lys751Gln, A/C), no remarkable differences for genotype distribution and allele frequencies were observed between BC group and control group in the study. The genotype frequency for homozygote A/A in XPD exon 6 (Arg156Arg, C/A) were significantly different between BC and control groups (P < 0.0001, odds ratio = 2.14; 95% confidence interval 1.44-3.17). The data indicate a possible role for XPD (Arg156Arg, C/A) polymorphisms in BC susceptibility. PMID- 18415713 TI - No strong association between HER-2/neu protein overexpression and gene amplification in high-grade invasive urothelial carcinomas. AB - The generation of urothelial carcinoma is caused by the accumulation of various molecular changes, as in most malignancies. There are conflicting data about the status of HER-2/neu oncogene in urothelial carcinomas. The aim of this study was to determine the status of HER-2/neu oncogene in high-grade invasive urothelial carcinoma of urinary bladder both in protein and DNA level. We evaluated HER 2/neu protein overexpression by immunohistochemistry (IHC) and gene amplification by fluorescent in situ hybridization (FISH) and real-time quantitative PCR in paraffin-embedded samples of high-grade invasive urothelial carcinoma obtained from 36 patients. Polysomy 17 was also assessed by FISH. Immunohistochemically, HER-2/neu protein overexpression was observed in 22 (61.1%) tumors (ten tumors with score 3+ and 12 with score 2+). Fourteen of 36 tumors (38.9%) were evaluated as negative (score 0 or 1+). Complete concordance between FISH and the PCR was seen in all of the samples scored as 0 and 1+ by IHC. HER-2/neu gene amplification was observed in three of 27 (11.1%) tumors by FISH (nine samples were non-informative) and in eight of 36 (22.2%) tumors by the PCR. The complete concordance between HER2-2/neu protein overexpression and gene amplification was seen only in three of 27 tumors. Polysomy 17 was seen in nine tumors (33.3%). The results indicated that, in contrast to breast cancer, there was no strong association between HER-2/neu overexpression and gene amplification in invasive urothelial carcinomas, and polysomy 17 was higher in tumors showing HER-2/neu overexpression. PMID- 18415714 TI - SORS: a universal one-round PCR-based method for site-directed mutagenesis. AB - We have developed a novel protocol for site-directed mutagenesis of double stranded DNA. The procedure, termed SORS (named because it undergoes the sequential procedure of segmentation-overhang creating PCR-reannealing-splicing) mutagenesis, is exemplified by a substitution, a deletion, and an insertion of nucleotide(s) in target genes. The template DNA is PCR-amplified into two separate segments divided at the prospective mutation site, and each segment is amplified in two parallel PCRs using primers introducing the mutation. The primers are designed to be able to create protruding bases upon pooling, denaturing, and reannealing the two parallel reactions. The protruding bases at the prospective junction of the two segments are mutually complementary; therefore, the two segments can be re-spliced together to generate the mutated gene. Compared to previously published protocols, this procedure is rapid, restriction-independent and ensures higher success rate and lower potential to produce second-site mutations. PMID- 18415715 TI - Sustained expression of human cytomegalovirus glycoprotein B (UL55) in the seeds of homozygous rice plants. AB - Production of recombinant subunit vaccines in transgenic plants may be a means of reducing vaccine costs while increasing availability and safety. A plant-derived product found safe and effective for oral administration would provide additional advantages when used as a vaccine. Outstanding issues with the technology include transgene stability through successive generations and consistent bioproduction. We previously reported expression of glycoprotein B (gB) of human cytomegalovirus in seeds of transgenic tobacco. Here the goal was to determine if gB could be similarly expressed in rice, and if so, to examine expression over several plant generations. Results show that immunoreactive gB was successfully expressed in transgenic rice seeds, with sustained expression over three generations. The gB contained several neutralizing epitopes and was stable over 27 months. PMID- 18415716 TI - Physiotherapy after traumatic brain injury: a systematic review of the literature. AB - PRIMARY OBJECTIVES: At present there are no standardized recommendations concerning physiotherapy of individuals with traumatic brain injury (TBI) resulting in a high variability of methods and intensity. The aim of this literature review is to develop recommendations concerning physiotherapy in the post-acute phase after TBI on the basis of scientific evidence. METHOD: literature review: data bases: PubMed, PEDro, OT-Seeker, Cochrane and Cinahl. KEYWORDS: brain injury (in PEDro, OT-Seeker, Cochrane), brain injury AND physical therapy (in PubMed and Cinahl). RESULTS: Fourteen studies met the inclusion criteria and were grouped into sub-groups: sensory stimulation, therapy intensity, casting/splinting, exercise or aerobic training and functional skill training. While for sensory stimulation evidence could not be proven, a strong evidence exists that more intensive rehabilitation programmes lead to earlier functional abilities. The recommendation due to casting for the improvement of passive range of motion is a grade B, while only a C recommendation is appropriate concerning tonus reduction. Strong evidence exists that intensive task-orientated rehabilitation programmes lead to earlier and better functional abilities. CONCLUSION: Although some recommendations for the effectiveness of physical therapy interventions could be expressed, there are many questions concerning the treatment of humans with TBI which have not been investigated so far. Especially on the level of activity and participation only a few studies exist. PMID- 18415717 TI - Depression in the elderly after traumatic brain injury: a systematic review. AB - PRIMARY OBJECTIVE: To systematically review recent research published in peer reviewed journals that focus on depression in the elderly with TBI. To better understand the prevalence of depression in the elderly after TBI and to note any differences or similarities between the elderly and their younger counterparts. Explore depression in the general elderly population and depression in the overall TBI population to better understand depression in the elderly after TBI. METHODS AND PROCEDURES: MEDLINE and PsychINFO searches garnered one peer-reviewed English language article that met the following inclusion criteria: elderly participants with TBI, depression directly assessed and use of self-report measures and/or structured clinical interview. MAIN OUTCOMES AND RESULTS: Prevalence of depression in the elderly range from 1.8-8.9% in community-residing elders to 25% in nursing homes and long-term care settings. Prevalence of depression in the overall TBI population ranges from 15.3-42%. Little research has been published on depression in the elderly following TBI, but the available research report a prevalence of 21-37%. CONCLUSION: The presence of depression after TBI in the elderly has important implications for rehabilitation treatment. There is a great need for more research in this area requiring more stringent age markers and methodology. PMID- 18415718 TI - Prevalence and types of sleep disturbances acutely after traumatic brain injury. AB - PRIMARY OBJECTIVE: To assess the prevalence of and risk factors for sleep disturbances in the acute post-traumatic brain injury (TBI) period. RESEARCH DESIGN: Longitudinal, observational study. METHODS AND PROCEDURES: Fifty-four first time closed-head injury patients were recruited and evaluated within 3 months after injury. Pre-injury and post-injury sleep disturbances were compared on the Medical Outcome Scale for Sleep. The subjects were also assessed on anxiety, depression, medical comorbidity and severity of TBI. MAIN OUTCOMES AND RESULTS: Subjects were worse on most sleep measures after TBI compared to before TBI. Anxiety disorder secondary to TBI was the most consistent significant risk factor to be associated with worsening sleep status. CONCLUSIONS: Anxiety is associated with sleep disturbances after TBI. Further studies need to be done to evaluate if this is a causal relationship. PMID- 18415719 TI - Cognitive and affective outcomes of more severe compared to less severe carbon monoxide poisoning. AB - PRIMARY OBJECTIVE: To assess cognitive sequelae, depression and anxiety following carbon monoxide (CO) poisoning. RESEARCH DESIGN: Prospective cohort study. METHODS AND PROCEDURES: This study prospectively followed 256 patients, 55 with less severe and 201 with more severe CO-poisoning. The prevalence of cognitive sequelae, depression and anxiety at 6 weeks, 6 months and 12 months was compared in patients with less severe and more severe CO-poisoning. MAIN OUTCOMES AND RESULTS: Of the less severe CO-poisoned patients, 39% had cognitive sequelae, 21% depression and 30% anxiety at 6 weeks. Of the more severe CO-poisoned patients, 35% had cognitive sequelae, 16% depression and 11% anxiety at 6 weeks. There was no difference in the prevalence of cognitive sequelae at any time. The prevalence of depression was higher in patients with less compared with more severe CO poisoning at 6 months (p = 0.04), but not 6 weeks or 12 months. The prevalence of anxiety was higher in patients with less compared to more severe CO-poisoning at 6 weeks (p = 0.008), but not 6 or 12 months. Anxiety decreased over time in the less severe group (p < 0.01). CONCLUSION: CO-related cognitive sequelae, depression and anxiety are common and may be independent of poisoning severity. PMID- 18415720 TI - Conversation-based assessment of social cognition in adults with traumatic brain injury. AB - PRIMARY OBJECTIVE: The purpose of this study was to characterize performance of individuals with traumatic brain injury (TBI) on a novel video-based test, the Video Social Inference Test (VSIT). The VSIT was designed to capture social inference processes that would be engaged in daily conversations. The test required both initial social inferences and also predictions or explanations of subsequent behaviours. RESEARCH DESIGN: Between-groups comparison. METHODS: Adults with TBI (n = 19) and typical controls matched for age and sex (n = 19) completed the VSIT, as well as a working memory test and the Eyes Test, a widely used picture-based test of social cognition. RESULTS: VSIT scores were lower in the TBI group and higher than on the Eyes Test. Participants in both groups had lower scores when required to predict or explain future behaviours based on an initial social inference. CONCLUSIONS AND IMPLICATIONS: The results suggest that conversation-based stimuli may yield unique and useful information about social cognition beyond the laboratory. PMID- 18415721 TI - Determinants of compliance with hand splinting in an acute brain injured population. AB - PURPOSE: Poor compliance with hand splinting treatment in the acute injury stage increases the risk of ongoing disability by affecting recovery and functioning. The aims of this study were to identify key predictors of splinting non compliance in acute traumatic brain injury (TBI) patients and to determine the suitability of the Westmead PTA scale for predicting splinting compliance in TBI. METHOD: Retrospective medical record review of all patients who were (1) admitted to a major adult trauma hospital in 2005-2006 and (2) flagged as having concurrent brain and upper limb injuries. Data extracted included demographic information, co-morbidity, injury mechanism, TBI severity, incidence and duration of agitation. Compliance data included loss, removal or agitation with the splint or brace. RESULTS: Of the 71 subjects, 39.5% (n = 28) were non-compliant with their splint or brace; 60.5% (n = 43) were compliant. The presence and duration of agitation were the strongest predictors of non-compliance with splinting (p = 0.001 and p = 0.003, respectively). CONCLUSION: PTA status at splint application does not accurately predict compliance with splinting. This highlights a specific clinical gap in the management of hand injuries against a background of agitation and cognitive impairment. PMID- 18415722 TI - A brief neuropsychological screening test battery for cognitive dysfunction in Brazilian multiple sclerosis patients. AB - PRIMARY OBJECTIVE: The cognitive profile of patients with Multiple Sclerosis in the Brazilian population is relatively unknown and no cognitive screening instruments have been developed for this population. The purpose of the present study was to develop a neuropsychological screening instrument battery to identify cognitive impairment in epidemiological research protocols in this country. METHODS AND PROCEDURES: Fifty-four patients with Relapsing-Remitting Multiple Sclerosis (RRMS) and 54 controls matched by sex, age and educational level were evaluated through a long and comprehensive neuropsychological test battery. MAIN OUTCOMES AND RESULTS: In comparison to the control group, 51.8% of the patients with RRMS presented some form of cognitive impairment. Three neuropsychological tests - Verbal Fluency, Rey Auditory Verbal Learning and Enhanced Cued Recall - presented an 80.6% sensitivity and 97.2% specificity in differentiating RRMS patients with cognitive deficits from paired control subjects. CONCLUSIONS: These three tests constitute a Brief Battery that represents a first Brazilian MS cognitive screening instrument. However, further validation study is necessary to achieve better normative scores in a larger sample and to establish feasibility of this battery. PMID- 18415723 TI - Predictors of positive growth after traumatic brain injury: a longitudinal study. AB - PRIMARY OBJECTIVE: To investigate long-term positive psychological growth in individuals with traumatic brain injury (TBI) and to relate growth to injury characteristics and early outcomes. RESEARCH DESIGN: Longitudinal study. METHOD AND PROCEDURE: Long-term follow-up of a group of TBI survivors recruited between 1991-1995. In 2004, 240 of the 563 original participants were invited to take part in a follow-up study. At follow-up, survivors completed the Positive Changes in Outlook Questionnaire (CiOP) along with a structured interview/questionnaire which permitted a Glasgow Outcome Scale (GOSE) score to be assigned. RESULTS: One hundred and sixty-five TBI survivors completed both questionnaire and CiOP. One hundred and three (62%) participants had suffered severe TBI, 24 (15%) moderate and 38 (23%) mild. Mean length of follow-up was 11.5 years post-injury (range 9 25 years). On the GOSE at follow-up, 43 (26%) had severe disability; 72 (44%) moderate disability; and 50 (30%) good recovery. Scores on the CiOP indicated positive psychological growth in over half of the sample, as evidenced by agreement with items such as 'I don't take life for granted anymore' and 'I value my relationships much more now'. CiOP total scores did not correlate with any injury or early outcome variables. However, at long-term follow-up there was a negative correlation between positive growth and anxiety and depression. CONCLUSION: Survivors of mild, moderate and severe TBI showed evidence of long term positive changes in outlook. PMID- 18415725 TI - The relationship between socio-economic status and lexical development. AB - The British Picture Vocabulary Scale, second edition (BPVS-II), a measure of receptive vocabulary, is widely used by speech and language therapists and researchers into speech and language disorders, as an indicator of language delay, but it has frequently been suggested that receptive vocabulary may be more associated with socio-economic status. Studies on this topic have produced conflicting results. This study reviews the literature and tests the hypothesis that receptive vocabulary is associated with socio-economic status. The BPVS-II was administered to 76 typically-developing children aged 4 to 11, classified according to deprivation category, in Edinburgh, Scotland. The results showed no significant correlation between the two measures. Error patterns within the results are examined to discover why there should be discrepancy between them and the findings of other studies. The interaction between word frequency and the items used in the BPVS-II is examined, and implications for the use of the BPVS II with all children of primary school age are discussed. PMID- 18415726 TI - Lexical mixing in the early productive vocabularies of Maltese children: implications for intervention. AB - Since norms for vocabulary acquisition in Maltese children do not yet exist, documentation of productive vocabulary acquisition may contribute to establishing a baseline of lexical development. Clinical implications may thus be derived. The current study is a small-scale investigation of the proportions of Maltese and English lexemes in the vocabularies of ten normally-developing Maltese children aged between 12 and 30 months. The participants were primarily exposed to Maltese within their immediate environments, while receiving indirect exposure to English. Outcomes of parental report and language sampling were analysed for evidence of a bilingual dimension in these children's productive vocabularies. Translation equivalents were reported on by parents, but negligible evidence of equivalents emerged in conversational language use. In contrast, lexical borrowings were both reported and sampled. A substantial proportion of English lexemes were reported by the parents in the absence of Maltese equivalents. PMID- 18415727 TI - Avoidance strategies in an exceptional child during unsuccessful reading performances. AB - Employing an interactional analysis and focusing on the complexity of meaning making, this study investigates the uses of several types of avoidance strategies generated by a language-disordered boy who is struggling with literacy. The results suggest that these avoidance strategies may function as compensatory adaptations that assist him in overcoming his literacy limitations so that he can still sustain effective social action even within contexts where his literacy difficulties are highlighted. There are both theoretical and practical implications for these findings. PMID- 18415728 TI - Applying eye movement miscue analysis to the reading patterns of children with language impairment. AB - This paper presents a research report on an investigation into the reading eye movements of a bilingual 10-year, 10-month old girl with language impairment secondary to Downs Syndrome. Eye movement miscue analysis (Paulson, 2000) was employed to evaluate the visual sampling and oral productions of this child as she read from complete texts presented in Spanish and English. Results are presented in relationship to misconceptions manifest in the clinical practice of intervention providers. Based upon the research and this data, readers do not fixate on every word and the miscues they produce are not the result of failing to fixate on the text. PMID- 18415729 TI - Expressive versus receptive language skills in specific reading disorder. AB - Despite ample research into the language skills of children with specific reading disorder no studies so far have investigated whether there may be a difference between expressive and receptive language skills in this population. Yet, neuro anatomical models would predict that children who have specific reading disorder which is not associated with movement or attention difficulties, would have lower receptive language skills than expressive. This study investigates the difference between expressive and receptive language skills in a sample of 17 children with specific reading difficulty aged between 7 and 12 years. They were administered a battery of two receptive and two expressive language measures. The results showed that as the neuro-anatomical model would predict, the children scored significantly lower on tests of receptive than on tests of expressive language skills. PMID- 18415730 TI - Embodiment in communication--aphasia, apraxia and the possible role of mirroring and imitation. AB - The role of embodiment in communication is attracting an increased interest. This interest is to some extent caused by hypotheses and findings concerning mirror neurons in macaques, that is, neurons that are activated by production as well as perception of, for example, a certain movement of action. Mirror neurons seem to provide a fairly simple mechanism for acting, perceiving, imitating and pantomime, which could be crucial to the development of human communication and language. A number of theories try to extend similar ideas in describing human embodied communication. Some of the consequences of these theories are: (1) the close relation between speech and gestures; (2) the close relation between speech/language and praxis; and (3) the reconsideration of the importance for communication of more automatized versus more controlled processing. The purpose here is to point to possible consequences for clinical research and therapy concerning language disorders. PMID- 18415731 TI - Rethinking agrammatism: factors affecting the form of language elicited via clinical test procedures. AB - Current approaches to assessing agrammatism use data from restricted contexts, such as picture description and story telling tasks. There is evidence in the conversation analysis literature to suggest that conversational grammar may differ markedly from the grammar of such elicited language samples. The disparity between conversational and test grammar suggests that it is possible for the form of an agrammatic utterance to be motivated by the context in which it occurs. Thus, behaviours previously considered to be symptoms of agrammatism may be adaptations to talking in different environments. This study analyses two distinctive test response forms made by two agrammatic speakers, and discusses whether they might be strategic adaptations to testing rather than symptoms of impairment. PMID- 18415732 TI - Evaluation of a novel technique for assessing speech discrimination in children. AB - Methods used to assess children's speech perception and recognition in the clinical setting are out of step with current methods used to investigate these experimentally. Traditional methods of assessing speech discrimination, such as picture pointing, yield accuracy scores which may fail to detect subtle perceptual difficulties. This paper will report a novel method of assessing speech input processing that uses measurement of children's eye movements to provide information on speed and confidence as well as accuracy in discriminating phonological contrasts. Participants were typically developing children aged 2-7 years. Pairs of pictures representing auditory minimal pairs which varied in type and degree of phonological contrast were presented on a computer screen while the child heard a word matching one of these pictures. The child's eye movements in response to these stimuli were videorecorded for subsequent analysis of duration and direction of gaze. The effects of age and stimulus type on eye gaze were examined. The results were compared with those of a traditional picture pointing task using the same stimuli. The informativeness of the novel technique is evaluated on the basis of the findings. PMID- 18415733 TI - Parental involvement in speech intervention: a national survey. AB - A survey of 277 speech language pathologists (SLPs) investigated beliefs and practice regarding parents' involvement in service planning and delivery for children with speech impairment. Although the SLPs frequently involved parents in service delivery for speech intervention, parental involvement in service planning was less frequent. SLPs working in educational settings involved parents to a lesser extent than SLPs working in health settings and private practice. More experienced SLPs involved parents less in decision-making. A gap between the SLPs' beliefs and practice was found, with stated beliefs not always reflecting practice. 40% of respondents were unhappy with the level of parental involvement and perceived workplace, personal and parental barriers to working effectively with parents. Although the SLPs indicated that they believed in and used family centered practices, beliefs and practice regarding parent decision-making were therapist- rather than family-centered. PMID- 18415734 TI - The contribution of polysyllabic words in clinical decision making about children's speech. AB - Poor polysyllabic word (PSW) production seems to mark paediatric speech impairment as well as impairment in language, literacy and phonological processing. As impairment in these domains may only manifest in PSWs, PSW production may provide unique information that is often excluded from clinical decision making because insufficient PSWs are included in speech tests. A 5-stage model of PSW acquisition is described. The model, grounded in optimality theory, expresses a reciprocal relationship between the relaxation of markedness constraints and the contraction of faithfulness constraints from 12 months of age to adolescence. The markedness constraints that persist to the age of 7;11 years are associated with non-final weak syllables and within-word consonant sequences. Output changes are argued to reflect increasing specification of phonological representations with age, liberating information for motor planning and execution, resulting in increasingly accurate output. The clinical implications of PSWs in assessment and therapy are discussed. PMID- 18415735 TI - Optimizing stimulus length for clinical nasalance measures in Swedish. AB - Standardized passages used for speech nasalance measures may be too long for clinical use with very young or non-compliant patients. The aim of this study was to establish whether nasalance scores from shorter sections of three Swedish speech stimuli were equivalent to those from their corresponding whole stimulus. Nasalance recordings for three Swedish speech stimuli (oral, nasal and oronasal) were obtained from 29 typically developing Swedish children (7-11 years). Cumulative sentence combinations were evaluated for equivalence to their respective whole passages according to two different criteria: one based on t testing and the other on cumulative frequencies of score differences. Results showed that shorter sequences of sentences could be considered equivalent to the whole passage for the oral and nasal stimuli, provided that the sentences were sequenced in order of increasing difference from the whole stimulus score and that those sentences with scores significantly different from the whole passage score were not included. PMID- 18415736 TI - Intonation features of the expression of emotions in Spanish: preliminary study for a prosody assessment procedure. AB - This study aimed to find out what intonation features reliably represent the emotions of "liking" as opposed to "disliking" in the Spanish language, with a view to designing a prosody assessment procedure for use with children with speech and language disorders. 18 intonationally different prosodic realisations (tokens) of one word (limon) were recorded by one native Spanish speaker. The tokens were deemed representative of two categories of emotion: liking or disliking of the taste "lemon". 30 native Spanish speakers assigned them to the two categories and rated their expressiveness on a six-point scale. For all tokens except two, agreement between judges as to category was highly significant, some tokens attracting 100% agreement. The intonation contours most related to expressiveness levels were: for "liking", an inverted U form contour with exaggerated pitch peak within the tonic syllable; and for "disliking", a flat melodic contour with a slight fall. PMID- 18415737 TI - Perception and confusion of speech sounds by adults with a cochlear implant. AB - The aim of this pilot study was to identify the most common speech sound confusions of 5 Norwegian cochlear implanted post-lingually deafened adults. We played recorded nonwords, aCa, iCi and bVb, to our informants, asked them to repeat what they heard, recorded their repetitions and transcribed these phonetically. We arranged the collected data in confusion matrices to find the most common and most uncommon speech sound confusions. We found that the voiced and unvoiced consonants are seldom confused. We also found that there was a higher rate of consonant confusion for the iCi words than for the aCa words. The most frequent confusion was [eta] perceived as [n], [m] perceived as [n] and [upsilon] perceived as [n]. For the consonants, manner of articulation was rarely confused, but place of articulation was often confused. An exception from this was the confusion of [l] and [n], which differs only in manner of articulation. The latter is in accordance with reports we get from clinicians. We postulate that this is caused by the speech processing of the cochlear implant. We found less confusion of the vowels, which can be explained by the fact that vowels have much higher energy and longer duration than most of the consonants. The most frequent confusion was [a:] perceived as [see text] and [u:] perceived as [see text]. [e:], [i:] and [see text] were never confused with other vowels. PMID- 18415738 TI - Data analysis of Chinese characters in primary school corpora of Hong Kong and mainland China: preliminary theoretical interpretations. AB - Metalinguistic awareness (an awareness about the structure of orthography) had been considered vital for reading acquisition. The awareness of phonological regularity and consistency had been found in advanced readers in recent research. Evidence based on simplified Chinese suggested the effect of semantic transparency on reading in school readers. Studies based on traditional Chinese also reported that reading acquisition, including the development of metalinguistic awareness, is affected by script, properties of characters in school curricula, approaches and strategies of reading training. This paper reports the comparison between corpora of simplified Chinese characters based on primary school textbooks and the updated Hong Kong Corpus of Primary School Chinese (HKCPSC). The proportion of characters in the total curriculum, the ratio of phonetic-semantic compounds, visual complexity (defined by the number of strokes) and the levels of phonetic regularity and semantic transparency of Chinese characters across grades in the two corpora are compared. Two marked differences found are the frequency-weighted proportion of regular characters and the proportion of semantically transparent characters across grades. The relationships between the data and recent findings of reading development in Chinese are discussed. PMID- 18415739 TI - The signs B and B-bent in Israeli sign language according to the theory of Phonology as Human Behavior. AB - The purpose of the present research is to examine which of the two factors: (1) the iconic-semiotic factor; or (2) the human-phonetic factor is more relevant in explaining the appearance and distribution of the hand shape B-bent in Israeli Sign Language (ISL). The B-bent shape has been the subject of much attention in sign language research revolving around the question of its status as a phoneme. The arguments supporting the phonemic status of the B-bent hand shape have been primarily based on the semiotic opposition between the hand shape B and the hand shape B-bent. It has been claimed that in Italian Sign Language the hand shape B is perceptually distinct from the hand shape B-bent, i.e. in opposition to the general, neutral, unmarked meaning of the hand shape B, the iconic hand shape B bent has a more narrow, specific and marked meaning: DELIMIT. The B-bent hand shape appears in spatial-temporal signs such as "a little before, ahead, postpone or behind". In these signs the iconic structure of the hand shape B-bent is utilized to mark borders in space and time. The arguments opposing the perceptual/phonemic distinction between these hand shapes is based on the human phonetic factor, i.e. the need to reduce the effort on the part of the wrist joints in specific phonetic environments. We performed a quantitative and qualitative content analysis of the distribution of the basic units of 560 lexical signs taken from a stratified random sample from the ISL dictionary. The results were analyzed in the framework of the sign-oriented linguistic theory of the Columbia School including the theory of Phonology as Human Behavior. Our data revealed that the B-bent hand shape--as all the "building blocks" of the ISL--is a morpho-phonemic unit. We found that there is not only a phonemic distinction between hand shape B and hand shape B-bent in ISL (based on minimal pairs), but there is also a perceptual distinction between them. The qualitative analysis shows that the distribution of the B and the B-bent shapes in the lexicon of ISL is not random but rather motivated by a specific iconic meaning attached to each one of them. The results of our research lead to the conclusion that iconicity is the dominant explanatory factor in the Phonology of ISL. PMID- 18415740 TI - Past tense morphology in Cri du chat syndrome: experimental evidence. AB - It has been observed that persons with Cri du chat syndrome (CDCS) have poor language production. However, very few studies have addressed the question whether all aspects of language production are equally afflicted, or whether there are differences between for instance phonological and morphological abilities. The present study was aimed at investigating to what extent persons with CDCS inflect verbs, and, if so, which inflectional patterns in the target language they make use of, and further what kinds of inflectional errors they make. We compared the performance of three subjects (aged 11, 15 and 22) with CDCS on a past tense elicitation task. We found that all three subjects inflected verbs. The number of correct responses, however, varied from 28% to 53%. The majority of errors were over-generalizations, imitation of input, no response and substitution by semantically related verbs. PMID- 18415741 TI - Compliance with guidelines for the management of acute pancreatitis: a protocol is not enough. PMID- 18415742 TI - Subjective results and symptomatic outcome after fundoplication revision. AB - OBJECTIVE: In a small proportion of patients, fundoplication fails and a reoperation is required. However, there are few reports on the symptomatic outcome after reoperative antireflux surgery. The aim of this study was to evaluate the results after fundoplication revision. MATERIAL AND METHODS: All patients (n=71) undergoing fundoplication revision between 1997 and 2005 were included in this study. The follow-up data were collected both from the hospital records and during postoperative control visits, including a personal interview using a structured questionnaire; follow-up was completed by 61 patients (88.4%). RESULTS: The primary fundoplications included both open (n=21) and laparoscopic (n=40) approaches; 92% (n=56) of the reoperations were open procedures. The morbidity rate was 21% and the reoperation rate 16%. Fifty-six percent (n=34) of the patients regarded the result of their reoperative surgery as excellent, good or satisfactory at a mean follow-up of 51 months; 66% of the patients had no significant reflux symptoms after re-fundoplication. With the benefit of hindsight, 77% of the patients would again choose to undergo re-fundoplication, but only 48% of the patients would again primarily choose surgical treatment. Mortality rate was 1.4% (n=1) and in three patients the reoperative treatment required total gastrectomy. CONCLUSIONS: These suboptimal results show that surgical treatment for gastro-oesophageal reflux disease in general is far from being perfect and this is even more marked after reoperative antireflux surgery, as fundoplication revision can result in severe complications. This emphasizes the importance of proper patient selection for both initial and reoperative antireflux surgery. PMID- 18415743 TI - Relevance of the detection of intestinal metaplasia in non-dysplastic columnar lined oesophagus. AB - OBJECTIVE: In the USA, detection of intestinal metaplasia is a requirement for enrollment in surveillance programmes for dysplasia or adenocarcinoma in columnar lined oesophagus. In the UK, it is believed that failure to detect intestinal metaplasia at index endoscopy does not imply its absence within the columnarized segment or that the tissue is not at risk of neoplastic transformation. The aim of this study was to investigate the factors predicting the probability of detection of intestinal metaplasia in the columnarized segment. MATERIAL AND METHODS: Demonstration of intestinal metaplasia was analysed in 3568 biopsies of non-dysplastic columnar-lined oesophagus from 1751 patients from 7 centres in the UK. Development of dysplasia and adenocarcinoma was analysed in 322 patients without intestinal metaplasia and compared with that in 612 patients with intestinal metaplasia. RESULTS: Intestinal metaplasia was more commonly detected in males than in females (odds ratio 1.244), longer segment length (10.3% increase per centimetre) and increasing number of biopsies taken (24% increase per unit increase). After 5 years of follow-up, 54.8% of patients without intestinal metaplasia at index endoscopy demonstrated intestinal metaplasia, and 90.8% after 10 years. There was no significant difference in the rate of development of dysplasia or adenocarcinoma between patients with or without intestinal metaplasia detection at index endoscopy. CONCLUSIONS: Detection of intestinal metaplasia is subject to significant sampling error. It increases with segment length and number of biopsies taken. In the majority of patients, if sufficient biopsies are taken over time, intestinal metaplasia will be demonstrated. The decision to offer surveillance should not be based upon the presence or absence of intestinal metaplasia at index endoscopy as the risk of dysplasia and adenocarcinoma is similar in both groups. PMID- 18415744 TI - Effects of acid antisecretory drugs on mucus barrier of the rat against 5 fluorouracil-induced gastrointestinal mucositis. AB - OBJECTIVE: Acid antisecretory agents are used for the prophylaxis of cancer chemotherapy (CT)-induced gastrointestinal (GI) mucositis. Although these drugs seem to be clinically beneficial, data on their effects on the GI mucosal defense during CT treatment are scant. The objective of this study was to compare the effects of omeprazole, lansoprazole, and lafutidine on mucin, a major mucus component, during 5-fluorouracil (5-FU) treatment, as a CT regimen. MATERIAL AND METHODS: Rats, weighing approximately 230 g, were divided into five groups. The control group was administered 0.5% carboxymethylcellulose orally once daily for 5 days. The second, third, fourth, and fifth groups were treated with 5-FU (50 mg/kg), 5-FU plus omeprazole (10 mg/kg), 5-FU plus lansoprazole (10 mg/kg), and 5 FU plus lafutidine (30 mg/kg) in the same way, respectively. The rats were sacrificed on the sixth day, and their stomachs and small intestines were removed. Using anti-mucin monoclonal antibodies, we compared the immunoreactivity in different areas of the rats' GI tracts as well as the mucin content. RESULTS: Body-weight decreased in rats in the 5-FU group. Lafutidine, but neither omeprazole nor lansoprazole, inhibited the 5-FU-induced weight loss. Mucosal damage and reduced mucin content in stomach and small intestine were observed in rats receiving 5-FU alone. In the stomach, all antisecretory drugs caused the protective effects against 5-FU-induced mucosal injury and alleviation of the decreased mucin accumulation. In the jejunum and ileum, lafutidine, but neither omeprazole nor lansoprazole, ameliorated the 5-FU-induced mucosal damage and decreased mucin accumulation. CONCLUSION: Lafutidine could offer the possibility of more effective prevention of CT-induced mucositis through the activation of GI mucus cells. PMID- 18415745 TI - Efficacy of near-infrared irradiation on intractable hiccup in custom-set acupoints: evidence-based analysis of treatment outcome and associated factors. AB - OBJECTIVE: The condition intractable hiccup (IH) is generally an incapacitating disorder indicating neurologic or non-neurologic disorders. Linearly polarized, near-infrared irradiation (SL) is shown to be effective in the treatment of IH where it is applied on custom-set acupoints. The aim of this study was to investigate the treatment efficacy of IH by SL on the acupoints and to survey the relationship between IH and comorbid-related factors. MATERIAL AND METHODS. A total of 35 patients with IH were enrolled prospectively and divided into central and non-central groups. All patients received SL using an SG-type lens unit on the relevant acupoints and were followed-up for up to 6 months for efficacy of the novel therapy. RESULTS: There was no significant difference between patients with IH produced by central or non-central origins (p=0.7105) regarding the therapeutic effect of SL; however, the effects of age, bed-shaking, gun-waving motion and nasogastric (NG)-tube placement were significant. The severity index of IH was analyzed and found to be associated with the seasons. For those patients with elevated levels of aspartate aminotransferase (ASAT) after therapy, it took a significantly shorter (p=0.0029) period of time to treat IH with this novel therapy (p=0.0029). Thirty-four patients had complete resolution of IH within a few days of beginning SL, with partial resolution in 1 patient only. CONCLUSIONS: Without potential side effects, SL on custom-set acupoints could be a complementary therapy for patients with IH regardless of central or non-central origins. PMID- 18415746 TI - Measurement of colonic transit using a delayed-release capsule containing radio opaque markers. AB - OBJECTIVE: Colonic transit scintigraphy has several advantages in comparison with radio-opaque marker (ROM) techniques but it is limited to specialized centers. The aim of the study was to validate the ROM technique for measuring colonic transit using delayed-release capsules. MATERIAL AND METHODS: Colonic transit is expressed as the sum of the weighted mean marker residence times in the colon (geometric centers). Delayed-release capsules containing radio-opaque markers (DC ROMs) or scintigraphic methods were compared simultaneously in 10 healthy volunteers. Reproducibility and responsiveness of DC-ROM measurements were evaluated in 10 healthy volunteers at an interval of 2 weeks and after bisacodyl treatment, respectively. RESULTS: The transit profiles between the scintigraphic and DC-ROM methods were highly correlated (r = 0.994, p < 0.01). The reproducibility and responsiveness of DC-ROM measurements were good. Colonic transit measured by DC-ROM correlated with the stool form scale (r = 0.66, p < 0.01). CONCLUSION: The DC-ROM test is simple, highly reproducible, responsive, and appropriate for clinical trials and clinical practice. PMID- 18415747 TI - Dietary alpha-ketoglutarate reduces gastrectomy-evoked loss of calvaria and trabecular bone in female rats. AB - OBJECTIVE: Surgical removal of the stomach (gastrectomy, Gx) leads to osteopenia in animals and in humans. In the rat, Gx adversely affects calvaria and trabecular bone. alpha-Ketoglutarate (AKG) is a precursor of hydroxyproline--the most abundant amino acid in bone collagen. The purpose of this study was to investigate the effects of dietary AKG on Gx-induced osteopenia. MATERIAL AND METHODS: Twenty female Sprague-Dawley rats were subjected to Gx and divided between two groups: Gx+AKG in the drinking water and Gx+Vehicle (i.e. drinking water without AKG). Another 20 rats were sham-operated and divided between two groups: Sham+AKG and Sham+Vehicle. The daily dose of AKG was 0.43 g per 100 g rat. All the rats were killed 8 weeks later and the calvariae, femora and tibiae were collected. The integrity of the calvariae was analysed planimetrically, following transillumination and photography. The bone mineral content (BMC) and bone mineral density (BMD) were measured in the right femorae and tibiae (bone densitometry), leaving the left femorae and tibiae to be analysed histomorphometrically (measurement of trabecular bone volume and trabecular fractal dimension). RESULTS: Gx caused calvarial bone degradation, reduced trabecular bone (femur and tibia) and impaired trabecular architecture. In addition, Gx lowered the femoral/tibial BMC and BMD (mainly cortical bone). Dietary AKG counteracted the Gx-evoked impairment of calvaria and trabecular bone but failed to affect the BMC and the BMD in either sham- operated or Gx rats. CONCLUSIONS: Gx resulted in loss of calvarial, trabecular and cortical bone in the rat. AKG counteracted the effect of Gx on calvaria and trabecular bone but not on cortical bone. PMID- 18415748 TI - Epithelial and stromal genetic instability linked to tumor suppressor genes in ulcerative colitis-associated tumorigenesis. AB - OBJECTIVE: We have previously documented not only epithelial but also stromal genetic instability in ulcerative colitis (UC)-associated lesions, including adenocarcinomas, using microsatellite markers close to the p53 gene on chromosome 17 (Chr.17). However, about half of the UC-associated tumors do not have p53 gene alterations. The purpose of this study was to detect early genetic instability (loss of heterozygosity (LOH) and microsatellite instability (MSI)) of both epithelial and stromal cells in UC-associated tumorigenesis, using different microsatellite markers from the p53 gene. MATERIAL AND METHODS: The laser captured microdissection-PCR-GeneScan method was applied to investigate genetic instability in both the epithelial and stromal elements of early UC-associated lesions (regenerative mucosa and dysplasia) and carcinomas using multiple microsatellite markers, chiefly close to tumor suppressor genes (TSGs: p16(INK4A), Rb, Smad4 and fragile histidine triad (FHIT)). Furthermore, expression of their gene products was analyzed by immunohistochemistry. RESULTS: In epithelium, although LOH for Chr.17 markers increased along with histological progression, the frequencies of LOH or MSI for TSG markers were found to be almost constantly increased in both stromal and epithelial components of all lesion types. In contrast, genetic instability of National Cancer Institute (NCI) recommended standard markers was not found to be significantly correlated with UC associated tumorigenesis. Immunohistochemically, epithelial p16(INK4A) expression tended to be decreased in LOH-positive lesions (p = 0.0780) and Smad4 expression was significantly decreased (p < 0.05). CONCLUSIONS: These results suggest that genetic instability in the stroma, especially regarding TSG markers, may play an important role in early-phase, UC-associated tumorigenesis. In addition, decreased expression of TSG due to genetic alteration might contribute to tumorigenesis. PMID- 18415749 TI - Acute gastroenteritis outbreaks in Catalonia, Spain: norovirus versus Salmonella. AB - OBJECTIVE: To describe the epidemiology of norovirus (NV) and Salmonella gastroenteritis outbreaks in Catalonia between 15 October 2004 and 30 October 2005. MATERIAL AND METHODS: Epidemiological data were obtained from outbreak reports. The aetiologic agent was investigated by culture for enteric bacteria, enzyme immunoanalyses, reverse transcription polymerase chain reaction (RT-PCR) and molecular epidemiology genotyping for NV. The chi(2) test was used to compare proportions. Incidence rates and their confidence intervals (CI) were calculated assuming a Poisson distribution. RESULTS: A total of 225 acute gastroenteritis (AG) outbreaks were reported, with 3436 persons affected and 115 hospitalized. Sixty outbreaks (27%) accounting for 1791 cases (52%) were caused by NV, with an attack rate (AR) of 34.7% and a hospitalization rate (HR) of 0.2%. Seventy- four outbreaks (33%) were caused by Salmonella spp., affecting 646 persons (18.8% (AR 45.6% and HR 13.7%)). The difference in AR between Salmonella spp. and NV was statistically significant (odds ratio (OR) 1.57; 95% CI: 1.39-1.78; p < 0.001). The median number of affected persons per outbreak was 3-fold higher for NV (15 (2-174)) than for Salmonella (5 (2-44)). The incidence rate of AG cases associated with NV outbreaks was significantly higher (24.9 per 100,000 persons/year (95% CI: 23.7-26.1)) than for those associated with Salmonella (5(2 44)) outbreaks (9.2 per 100,000 persons/year (95% CI: 8.5-9.9)). CONCLUSIONS: The disease burden and impact of NV highlight the importance of timely notification and investigation of outbreaks and the development and implementation of preventive measures, such as health and safety education for caregivers and food handlers in order to reduce the incidence of infections. PMID- 18415750 TI - Mortality from gastric cancer in patients followed with upper gastrointestinal endoscopy. AB - OBJECTIVE: Although commonly practiced in Japan, the effectiveness of regular screening with upper gastrointestinal (UGI) endoscopy against gastric cancer has not been well evidenced. The aim of the study was to investigate if gastric cancer-related mortality can be reduced by regular endoscopy. MATERIAL AND METHODS: The medical records of 833 patients with gastric ulcer (GU) and 2547 without ulcer (NU) were analyzed; these patients received long-term, repeated endoscopic examinations between 1969 and 2004. Gastric cancer incidence, death by gastric cancer, and overall survival were compared with those in a Japanese general population by calculating the standardized incidence ratios (SIRs) and standardized mortality ratios (SMRs). RESULTS: The interval between UGI endoscopic examinations was 1.4 +/- 1.4 years in the GU group and 1.8 +/- 1.5 years in the NU group. During follow-up, 32 patients with GU and 61 in the NU group developed gastric cancer, showing annual incidence rates of 0.40% (95% CI: 0.24-0.56%) and 0.38% (0.28-0.48%), and SIRs of 2.21 (1.44-2.98) and 1.72 (1.29 2.15), respectively. The 5-year survival rate exceeded 80% among patients who developed gastric cancer. SMRs for gastric cancer and overall deaths were 0.50 (0.01-0.99) and 1.05 (0.87-1.23) in GU patients, and 0.45 (0.15-0.74) and 0.78 (0.69-0.88) in NU patients. There were no significant differences between the two groups in gastric cancer incidence, mortality from gastric cancer, and overall survival. CONCLUSIONS: Mortality from gastric cancer could be reduced by regular UGI endoscopy in a population with a high incidence of gastric cancer. PMID- 18415751 TI - Long-term survivors among Danish patients after resection for ductal adenocarcinoma of the pancreas. PMID- 18415752 TI - Relevance of endotoxin receptor CD14 and TLR4 gene variants in chronic liver disease. AB - OBJECTIVE: The lipopolysaccharide (LPS)-triggered release of inflammatory cytokines from Kupffer cells is mediated via the CD14/TLR4 receptor complex. This inflammatory pathway can be influenced by alterations in genes encoding for LPS receptor components. Thus, a -260 C>T transition in the CD14 promoter is thought to result in enhanced CD14 expression thereby increasing the LPS responsiveness in chronic liver diseases, whereas a D299G exchange in the TLR4 gene has the opposite effect. Our objective was to analyze these two variations. MATERIAL AND METHODS: The study comprised 1712 patients with chronic liver diseases of different etiologies and 385 healthy controls. Genotyping was carried out by melting curve analysis with fluorescence resonance energy transfer (FRET) probes in the LightCycler. RESULTS: Genotype frequencies of CD14 -260C>T and TLR4 D299G did not significantly differ between patients and controls (CD14 TT 21.6% versus 21.8%; TLR4 DG or GG 9.7% versus 10.4%). We found no significant correlation of these alterations with disease course either in the groups of patients with alcoholic liver disease or hepatitis C virus (HCV) infection or among patients requiring liver transplantation. A significantly higher frequency of the CD14 260TT genotype was observed (36.6% versus 21.8% in healthy controls, p=0.036) only in a small subgroup of patients (n=41) with mild cryptogenic chronic liver disease. CONCLUSIONS: Variants within these LPS receptor genes were equally distributed among patients with chronic liver diseases of different etiologies and obviously do not confer an increased risk for the severity of these chronic liver processes. PMID- 18415753 TI - Percutaneous cholecystostomy: a bridge to surgery or definite management of acute cholecystitis in high-risk patients? AB - OBJECTIVE: Cholecystectomy is the standard treatment for acute cholecystitis, but in high-risk patients with serious comorbidity and in patients of advanced age there is substantial morbidity and mortality associated with the intervention. In these selected patients, percutaneous cholecystostomy (PCS) is an alternative mode of management. The aim of the present study was to evaluate the outcome of PCS in selected patients with acute cholecystitis. MATERIAL AND METHODS: Thirty five patients, representing 0.6% of all acute cholecystitis patients managed during the period 1994-2003, were subjected to PCS. Patients' charts were reviewed retrospectively for age, gender, comorbidity, hospital stay, procedure, complications and final outcome, including requirement of additional interventions. RESULTS: PCS was considered successful in 34/35 patients, 26 of whom responded within 3 days. Two patients required additional cholecystectomy 3 days and 20 months, respectively, after the PCS procedure. Two patients underwent endoscopic retrograde cholangiopancreatography (ERCP) and one patient underwent rotation lithotripsy. Four patients suffered recurrent biliary complaints after the acute episode of cholecystitis, while the only serious procedure-related complication was bile leakage from the gallbladder in one patient, which necessitated cholecystectomy. CONCLUSIONS: PCS is a comparatively safe and efficient procedure in the treatment of acute cholecystitis in high-risk patients with serious comorbidity and in elderly patients, contraindicating the general anaesthesia required for laparoscopic or open cholecystectomy. PMID- 18415754 TI - Usefulness of penicillamine-stimulated urinary copper excretion in the diagnosis of adult Wilson's disease. AB - OBJECTIVE: Diagnosis of Wilson's disease (WD) is reliant on liver biopsy (LB) and measurement of hepatic copper. The aim of this study was to determine the usefulness of penicillamine-stimulated urinary copper excretion (PS-UCE), a non invasive diagnostic test, for the diagnosis of WD in adults. MATERIAL AND METHODS: In this prospective study of patients with suspected WD, total serum copper, ceruloplasmin, basal 24-h UCE and PS-UCE levels were measured. LB with copper determination was performed in those patients with persistent hypertransaminasemia and low ceruloplasmin or basal UCE > 40 microg/24 h. Diagnosis was established if the ceruloplasmin level was found to be < 20 mg/dl and hepatic copper > 250 microg/g. Results. A total of 115 patients were studied; LB was performed in 43, and WD was diagnosed in 6 (13.9%). Significant differences between WD and non-WD patients were found for basal UCE (WD: median 134.3 microg/24 h versus non-WD: median 19.0 microg/24 h (p < 0.05)) and PS-UCE (WD: median 1284.0 microg/24 h versus non-WD: median 776.0 microg/24 h; p < 0.01). In the ROC (receiver-operated curve) analysis, PS-UCE was the best discriminant between WD and non-WD (area under the curve (AUC) = 0.911, best cut off point 1057 microg/24 h, 100% sensitivity, 82.3% specificity). CONCLUSIONS: PS UCE is probably a useful non-invasive test in the diagnosis of WD, improving the selection of patients for diagnostic liver biopsy. Patients with PS-UCE under 1057 microg/24 h only rarely will suffer from WD and are unlikely to benefit from LB. PMID- 18415755 TI - Histopathology of liver biopsies from a thiopurine-naive inflammatory bowel disease cohort: prevalence of nodular regenerative hyperplasia. AB - OBJECTIVE: Nodular regenerative hyperplasia (NRH) and sinusoidal dilatation have been described in relation to thiopurine use in patients with inflammatory bowel disease (IBD). However, there is a dearth of data on the prevalence of these histological abnormalities in general. The aim of our study was to describe the prevalence of these histological liver changes in a thiopurine-naive IBD cohort. MATERIAL AND METHODS: Liver biopsy specimens were obtained from patients who were treated in a referral center and who underwent gastrointestinal surgery for IBD. Patients were excluded if thiopurines were ever used. The liver specimens were pathohistologically assessed with special attention to NRH. RESULTS: A total of 83, properly stained, liver specimens (Crohn's disease 61%) were evaluated. NRH was observed in 6% compared to sinusoidal dilatation of varying degree in 34% of specimens. An older age at biopsy was correlated with NRH (p=0.015). Fibrosis and steatosis of varying degrees were detected in 31% and 36% of liver biopsies, respectively. No cases of liver cirrhosis were observed. CONCLUSIONS: Pathohistological hepatic abnormalities are common in non-thiopurine using IBD patients. The association between thiopurine use, NRH and sinusoidal dilatation may be weaker than as reported in recent literature, as there is relatively high background prevalence in selected series. PMID- 18415756 TI - Appropriate steroid therapy for autoimmune pancreatitis based on long-term outcome. AB - OBJECTIVE: Because autoimmune pancreatitis (AIP) responds well to corticosteroids, many AIP patients are given this treatment. However, there is no consensus on the indications, dose, or duration of steroid treatment. The aim of this study was to establish the most appropriate steroid therapy regimen. MATERIAL AND METHODS: We retrospectively reviewed morphological and serological improvement after steroid therapy and long-term outcome including relapse in 41 AIP patients who were given steroid therapy and were prospectively followed-up for more than 1 year. RESULTS: All patients responded to steroid therapy, which was given because of bile duct stenosis secondary to sclerosing cholangitis in 34 AIP patients. Pancreatic enlargement normalized within one month; however, 13 patients had incomplete resolution of pancreatic duct narrowing, and 14 patients had incomplete resolution of bile duct stenosis. There was no correlation between the degree of morphological improvement and the initial prednisolone dose (30 mg and 40 mg/day). In 58% of 19 patients, serum IgG4 elevation failed to normalize. Glucose intolerance improved in 38% of the 21 patients with diabetes mellitus. Nine patients who had complete morphological and serological resolution, stopped their medication, and none have relapsed. Thirty-two patients continued maintenance therapy, and 4 of these patients suffered relapse. CONCLUSIONS: The indications for steroid therapy in AIP patients include bile duct stenosis caused by sclerosing cholangitis and other systemic diseases, such as retroperitoneal fibrosis and diabetes mellitus. We recommend that oral prednisolone be used at an initial dose of 30 mg/day; maintenance therapy is required in cases without complete morphological and serological resolution. PMID- 18415757 TI - Risk factors for recurrent acute alcohol-associated pancreatitis: a prospective analysis. AB - OBJECTIVE: In an earlier retrospective study we showed that 46% of patients with acute alcoholic pancreatitis had recurrent attacks within 10-20 years, about 30% having a recurrence during the first 3 years. The aim of this prospective follow up study was to determine the risk factors associated with recurrences. MATERIAL AND METHODS: Sixty-eight patients, (59 M, 9 F, mean age 46 years, range 25-71 years) who survived their first acute alcohol-induced pancreatitis from January 2001 to January 2004 volunteered to participate in the study. The diagnostic criteria for acute pancreatitis were epigastric pain, serum amylase > 3 times the upper normal range, elevated serum C-reactive protein (CRP), and signs of acute pancreatitis in imaging. Other etiologies were excluded. Alcohol consumption and dependency were detected by the Alcohol Use Disorders Identification Test (AUDIT) and the Short Alcohol Dependence Data (SADD), respectively, and by attempting to evaluate recent use in grams of pure alcohol. Social and demographic data of the patients, smoking, body mass index, and the severity of the pancreatitis were recorded. Serum and fecal markers of the endocrine and exocrine function and secretin-stimulated MRCP were studied. The patients were followed for a median 38 (25-61) months for recurrences, and at the 2-year time-point had a follow-up visit to investigate any changes in alcohol consumption. RESULTS: Seventeen (25%) patients had recurrences of acute alcoholic pancreatitis during the follow-up. Pre-illness alcohol consumption, the severity of the pancreatitis, patient's social or demographic data, pancreatic function tests or morphologic changes in MRCP, or smoking did not correlate with recurrence. None of the 13 patients with consistent total abstinence from alcohol at 2 years developed recurrent pancreatitis compared with 17 out of 51 (33%) patients with at least some alcohol consumption (p = 0.02). Use of other sedatives than alcohol before the first attack of pancreatitis was an independent risk factor associated with recurrence (HR = 6.95, 95% CI 2.45-19.72, p < 0.001). A lower reduction in dependency on alcohol (less decreased SADD points) during 2 years was associated with a higher recurrence rate (HR = 0.921/each reduced point, 95% CI 0.872-0.974, p=0.004). CONCLUSIONS: Contrary to chronic pancreatitis, smoking was not found to be a risk factor for recurrent episodes after the first attack of acute alcoholic pancreatitis. Abstinence from alcohol protects against recurrent pancreatitis. Patients who developed recurrent acute pancreatitis had increased dependency on alcohol, demonstrated by the use of other sedatives in addition to alcohol and supported by the less decreased dependency during the follow-up. PMID- 18415758 TI - Colonic lavage with two polyethylene glycol solutions prior to colonoscopy makes no difference: a prospective randomized controlled trial. AB - OBJECTIVE: It is suggested that bowel preparations for colonoscopy are easier to tolerate when a smaller volume of solution with a more pleasant taste is used. The aim of this study was to establish equivalence between a 3-l sulphate-free polyethylene glycol solution (SF-PEG) and a 4-l PEG solution in effectiveness, patients' acceptability and tolerability. MATERIAL AND METHODS: The study comprised 110 patients scheduled for elective colonoscopy and randomized to receive either SF-PEG or PEG. Before colonoscopy, the patients completed a questionnaire on stool frequency, medication, concomitant diseases, the amount of solution ingested, willingness to re-take it, volume of other fluid taken and tolerance of bowel preparation, taste of the laxative and occurrence of abdominal cramps. Three experienced endoscopists, blinded to the type of preparation, assigned bowel-cleansing scores using a validated 5-point scale to assess cleansing effect. RESULTS: Data were available for 102 patients (44 M (40%), mean age 53 years, range 23-83 years). No significant differences were found in cleansing the rectosigmoid (p = 0.71) or complete colon (p = 0.79). Diverticulosis, constipation, gender and body mass index (BMI) did not influence cleansing. There was no significant difference in compliance between the two groups (p = 0.61). No differences were found for tolerance, taste and abdominal cramps. Patients who received SF-PEG had a preference for the same preparation next time in comparison with patients who had PEG cleansing (17 (33%) versus 4 (8%), respectively) (p = 0.03). CONCLUSIONS: Both preparations are comparable in their cleansing effect and toleration. However, patients prefer cleansing with a smaller volume of solution. Improving the acceptability of colonic preparation could improve willingness to undergo colonoscopies in the future. PMID- 18415759 TI - Quality of life is impaired in patients with peristomal bulging of a sigmoid colostomy. AB - OBJECTIVE: Peristomal bulging caused by hernia or prolapse is common in patients with a sigmoidostomy. It is not known whether and to what extent peristomal bulging influences various daily activities. The purpose of this study was to evaluate the effects of bulging by using a general and disease-specific health scale (Short Health Scale, SHS) and a stoma-specific quality of life (Stoma-QoL) questionnaire in patients with and without peristomal bulging. MATERIAL AND METHODS: Seventy patients with sigmoidostomies were examined to identify peristomal bulging. The mean (SD) age was 71.7 (13.7) years and the patients had had their sigmoidostomies for a mean of 8.1 (7.9) years. Bulging was noticed in 46 patients (66%) while 24 had no bulging. RESULTS: It was found that patients with bulging were at a disadvantage. In the SHS, patients with bulging reported significantly impaired QoL in 3 out of 4 scales regarding symptom load, worry and general sense of well-being. Also, in the Stoma-QoL questionnaire there was a significant difference between patients with and those without bulging. CONCLUSIONS: QoL evaluated with a general and disease-specific instrument (SHS) was significantly impaired in patients with bulging around a sigmoidostomy. The Stoma-QoL questionnaire showed a small but statistically significant difference between patients with and those without bulging but the clinical significance is uncertain. Further studies are required to evaluate the role of some of the individual items in the Stoma-QoL questionnaire. PMID- 18415760 TI - Spontaneous splenic rupture precipitated by cough: a case report and a review of the literature. AB - Spontaneous rupture of a normal spleen without a history of trauma is a rare clinical entity. We report on a case of atraumatic splenic rupture in a 61-year old man who presented to the emergency department for abdominal pain and hypotension. There was no evidence of hematologic or infectious disease involving the spleen. The chronic cough described by the patient was the main trigger for the rupture. Although, spontaneous splenic rupture is rare, it is vital that physicians consider this diagnosis when evaluating patients with abdominal pain and hypotension. Failure to consider splenic rupture could be catastrophic and early diagnosis is essential for a better outcome. PMID- 18415761 TI - Increased colonic nitric oxide level in active IBD. PMID- 18415762 TI - A combination of rectal beclomethasone diproprionate and mesalazine in ulcerative proctitis. PMID- 18415763 TI - Management of gout. PMID- 18415765 TI - Using SELDI-TOF MS to identify serum biomarkers of rheumatoid arthritis. AB - OBJECTIVES: No satisfactory biomarkers are currently available to screen for rheumatoid arthritis (RA). We have developed and evaluated surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF MS) for detection and analysis of multiple proteins for distinguishing individuals with RA from control individuals. METHODS: A total of 156 serum samples from 90 RA patients, 30 patients with ankylosing spondylitis (AS), and 36 healthy individuals were examined by SELDI technology. Spectral data were analysed by the support vector machine (SVM) approach and potential biomarkers were chosen for system training and were used to construct a diagnostic model. RESULTS: Pattern 1, consisting of four protein peaks with m/z values of 3899, 4594, 7566, and 13,842, distinguished RA from the healthy samples with sensitivity of 90.0% and a specificity of 91.7%. Pattern 2, consisting of m/z peaks 4287 and 6471, distinguished RA from AS with a sensitivity of 86.7% and a specificity of 85.0%. CONCLUSION: The combination of SELDI-TOF MS and SVM could facilitate the discovery of better biomarkers for RA and also provide a useful tool for molecular diagnosis in the future. PMID- 18415764 TI - Combination drug strategy in recent-onset rheumatoid arthritis suppresses collagen I degradation and is associated with retardation of radiological progression. AB - OBJECTIVE: To assess whether serum C-terminal cross-linking telopeptide of type I collagen (ICTP), a marker of type I collagen degradation, has any additional value in the assessment of treatment effect in patients with early rheumatoid arthritis (RA). METHODS: A total of 182 patients were randomized to treatment either with three disease-modifying anti-rheumatic drugs (DMARDs) and low-dose prednisolone (COMBI) or with a single DMARD with or without low-dose prednisolone (SINGLE). We investigated the prognostic value of serum ICTP level for the progression of joint destruction in X-rays (Larsen's score) from baseline to 2 years. RESULTS: There was a statistically significant decrease in serum ICTP levels from baseline to 1 year. At 6 months, the serum ICTP level was lower in the COMBI patients compared to that of the SINGLE cases (p = 0.008, after adjustment for baseline ICTP). When grouping the patients according to serum ICTP tertiles at 6 months, there was a statistically significant trend for increasing median change in Larsen score from baseline to 2 years from lowest to highest ICTP tertile in the SINGLE patients [p = 0.008, after adjustment for 28-joint Disease Activity Score (DAS28) score and RF status at baseline], while in the COMBI, the change remained low in all ICTP tertiles. CONCLUSIONS: The COMBI strategy for recent-onset RA results in early suppression of type I collagen degradation, which is reflected in radiological joint damage at 2 years. Serum ICTP at 6 months may be useful for identifying those RA patients whose treatment should be intensified to prevent further joint damage. PMID- 18415766 TI - Increased levels of autoantibodies against catalase and superoxide dismutase associated with oxidative stress in patients with rheumatoid arthritis and systemic lupus erythematosus. AB - OBJECTIVE: To evaluate the level of autoantibodies against superoxide dismutase (SOD) and catalase (CAT) in the sera of rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE) Tunisian patients, to study the oxidative profile among the same patients and to establish a correlation between the two parameters in order to understand the role of each one in the genesis of the two diseases. METHOD: Using a standard enzyme-linked immunosorbent assay (ELISA), the levels of immunoglobulin G (IgG) and IgM directed against CAT and SOD in the sera of 39 RA patients, 40 SLE patients, and 50 control healthy individuals were evaluated. The oxidative/antioxidative profile was tested by measuring serum malondialdehyde (MDA), conjugated dienes (CD), CAT activity, and SOD activity. RESULTS: Our data showed increased levels of IgG antibodies (Ab) against CAT in both groups of patients (p<0.05) compared to control subjects. However, the SLE patients displayed an increased level of anti-SOD IgG (p<0.05). In all patients the lipid peroxidation was confirmed by high levels of MDA and conjugated dienes (p<0.05). RA patients exhibited an increasing CAT and SOD activity in their sera (p<0.05) with a positive correlation observed between CAT and IgG anti-CAT (p<0.05). The same results were observed for SLE patients. In addition, a positive correlation was observed between anti-CAT Ab and anti-SOD Ab in SLE patients (p<0.05). CONCLUSION: Collectively, these results suggested that the primary factor causing the oxidative stress observed in RA and SLE is excessive free radical production rather than impaired CAT or SOD activity due to autoantibody inhibition. PMID- 18415767 TI - Hand function tests are important and sensitive tools for assessment of treatment response in patients with rheumatoid arthritis. AB - OBJECTIVES: To assess the usefulness of hand function measurements in a study of treatment effects of tumour necrosis factor (TNF) blockers and to define the relationship between different hand function tests and also relate hand function to general disability and disease activity. METHODS: The study group consisted of 49 patients with established rheumatoid arthritis (RA) who were followed for 1 year while on TNF inhibitors. Evaluation of hand function included Signals of Functional Impairment (SOFI), grip and pinch grip force, and the Grip Ability Test (GAT). General disability was assessed by the Health Assessment Questionnaire (HAQ) and disease activity by the 28-joint Disease Activity Score (DAS28). The standardized mean response (SMR) method was used to evaluate sensitivity to change for all hand tests using DAS28 and HAQ as external indicators of change. RESULTS: HAQ, DAS28, grip and pinch grip force, and GAT showed a highly significant improvement over time (p<0.001). The improvement in SOFI was also significant (p<0.01). The correlations between the different hand tests varied between 0.45 and 0.72. All hand function tests were significantly related to HAQ but showed only weak correlations to DAS28. SOFI, grip force, and pinch grip force showed large sensitivity for improvement in DAS28 and HAQ (SMR = 0.8-0.9). GAT showed modest sensitivity (SMR = 0.6-0.7). CONCLUSIONS: Patients with advanced RA attained considerable improvement in hand function that was only partly reflected by measures of general disability and disease activity. Focused assessment of hand function is therefore important for optimal evaluation of treatment response. PMID- 18415768 TI - Prevalence survey of rheumatoid arthritis and spondyloarthropathy in Lithuania. AB - OBJECTIVE: To assess the prevalence of rheumatoid arthritis (RA) and spondyloarthropathy (SpA) in two Lithuanian cities, Vilnius and Kaunas. METHODS: The first step in this study involved the translation and validation of a telephone questionnaire developed by rheumatologists and epidemiologists in France. The second step comprised the prevalence survey. To detect RA and SpA cases in the populations of Vilnius and Kaunas, 6542 subjects selected randomly (every 50th) from the latest telephone book were interviewed by telephone using a validated case detection questionnaire (the screening phase). All subjects with rheumatic symptoms but an uncertain diagnosis were contacted by a rheumatologist (confirmation phase) by telephone. If the diagnosis remained uncertain, the subjects were invited for a rheumatological examination. RESULTS: We attempted to contact 3370 telephone numbers in Vilnius and 3172 in Kaunas, and had a response rate of 62.5% and 67.7%, respectively. Over the course of all the study phases (telephone interview, rheumatologist's interview, and clinical examination), 39 RA cases and 27 SpA cases were detected, resulting in a crude prevalence of 0.92% for RA (95% CI 0.65-1.25) and 0.64% (95% CI 0.42-0.92) for SpA. The standardized prevalence rate according to age and sex in the Lithuanian population showed an RA prevalence of 0.55 (95% CI 0.39-0.74) and a SpA prevalence of 0.84 (95% CI 0.53-1.21). CONCLUSIONS: The prevalence of RA and SpA in Lithuania was found to be one of the higher rates in Europe. A telephone interview using a validated short questionnaire enabled a cost- and time-saving epidemiological survey to be conducted to detect RA and SpA cases in the community. PMID- 18415769 TI - Ankylosing spondylitis in Danish and Norwegian twins: occurrence and the relative importance of genetic vs. environmental effectors in disease causation. AB - OBJECTIVE: To estimate the influence of genetic effects in the aetiology and pathogenesis of ankylosing spondylitis (AS). METHODS: The study comprised one Norwegian and two Danish nationwide twin surveys. In 1994 and 2002, respectively, 37,388 and 46,331 Danish twin individuals were asked by questionnaire if they had AS. Similarly, in 1998, 12,718 Norwegian twins were asked if they had AS using a questionnaire phrased according to the Danish survey. Twins reporting AS were categorized according to the modified New York criteria. RESULTS: A total of 113 twin individuals reported AS, of whom 81 (72.3%) participated in validation of the diagnosis. After validation, 39 probands were diagnosed with AS. Subsequent invitation of co-twins resulted in 27 complete pairs. The point prevalence and the annual incidence of AS was 0.1% and 3/100,000 person-years (pyr) among the Danish twins. The positive predictive value of self-reported AS was 49.3%. Probandwise concordance rates on AS were (2/5) 40% in monozygotic (MZ) and (1/23) 4% in dizygotic (DZ) twins [difference 35% (95% CI 2.9-72.8), p = 0.26]. Heredity analysis including previously published and the present HLA-B27-positive twin pairs indicated that additive genetic effects account for 94% (95% CI 0.56-0.99) of the variance in the causation of AS. CONCLUSION: Self-reported AS needs careful validation. The occurrence of AS in a Danish twin population was 0.1% and accords well with previous studies on singletons in hospital settings. The present study adds to previous evidence of a major genetic effect in the pathogenesis of AS. PMID- 18415770 TI - Incidence of thromboembolic events in patients with primary Sjogren's syndrome. AB - Primary Sjogren's syndrome (pSS) is a connective tissue disease with symptoms and serological findings often overlapping with systemic lupus erythematosus (SLE) (1). Thromboembolic events are common in SLE but not in pSS (2)(3). However, case reports have described pSS patients who developed fulminant multiorgan disease due to thrombotic diathesis 4, and we have presented a case with acute catastrophic anti-phospholipid syndrome (APS) in a pSS patient (5). In this study we wanted to examine the incidence of thromboembolic episodes and relate these to the presence of autoantibodies and coagulation abnormalities in 90 pSS patients during a 4.6-year follow-up. PMID- 18415771 TI - Intravenous cyclophosphamide in patients with chronic systemic inflammatory diseases: morbidity and mortality. AB - BACKGROUND: Few data exist concerning the development of malignancies and haemorrhagic cystitis in patients with systemic autoimmune diseases previously treated with intravenous (iv) cyclophosphamide (CYC). The use of mesna prophylaxis is also controversial. METHODS: The medical records of all patients with chronic systemic inflammatory diseases treated with iv or oral CYC at Stavanger University Hospital from 1985 to 1999 were reviewed. Eighty-five patients were identified, of whom 75 patients had been treated with iv CYC only and were thus included in this study. Of these 75 patients, 20 (27%) had died and 55 (73%) were alive. Forty-two (76%) out of the 55 patients consented to undergo a comprehensive clinical examination, including a cystoscopy in 33 of them. The medical history of the patients not clinically examined was based solely on medical records. Data from the Cancer Registry of Norway and Statistics Norway were used for comparison with normative data in the general population. RESULTS: Six patients (8%) developed malignant disease compared with an expected number of 4.5, giving a standard incidence ratio of 1.5 [95% confidence interval (CI) 0.7 3.2]. The observed number of deaths was 23 compared to an expected number of 6.3, giving a standard mortality ratio of 3.7 (95% CI 2.4-5.5). CONCLUSIONS: The standard incidence ratio of cancer following iv CYC was increased, although not statistically significantly. No urinary bladder cancer or haemorrhagic cystitis developed even though mesna prophylaxis was not given. PMID- 18415772 TI - Soluble tumour necrosis factor receptors sTNFR1 and sTNFR2 are produced at sites of inflammation and are markers of arthritis activity in Behcet's disease. AB - OBJECTIVE: We analysed the production of soluble tumour necrosis factor receptors sTNFR1 and sTNFR2 at sites of inflammation and measured their plasma concentrations to evaluate them as biological markers of disease activity. METHODS: Plasma samples of 35 patients with Behcet's disease (BD) were collected prospectively at monthly intervals and grouped for inactive disease, active BD without arthritis, and active BD with arthritis. sTNFR1 and sTNFR2 concentrations were measured using immunoassays and compared with other biological disease activity parameters. Plasma sTNFR levels were compared to synovial fluid (SF) levels in seven patients. Sixteen tissue samples of mucocutaneous lesions were stained for TNFR2 expression by immunohistochemistry. RESULTS: sTNFR1 and sTNFR2 were found at increased plasma concentrations in active BD, with the highest concentration in active BD with arthritis (p<0.001). Concentrations of both sTNFRs were at least three times higher in SF of arthritic joints than in the corresponding plasma samples (p = 0.025). A change of more than 1 ng/mL of sTNFR2 plasma concentrations correlated with a concordant change in arthritic activity (96% confidence interval). Sensitivity to change was superior to that of sTNFR1, and other biological disease activity parameters such as erythrocyte sedimentation rate (ESR), immunoglobulin (Ig)G, IgA, and interleukin (IL)-10 plasma concentrations. A strong staining for TNFR2 was found in mucocutaneous lesions, where mast cells were identified as the major source for this receptor. CONCLUSIONS: This longitudinal study demonstrates that sTNFR2 plasma concentrations are closely linked with active BD, and especially with arthritis. Taken together with the expression of TNFR molecules in mast cells of mucocutaneous lesions, our results indicate a fundamental role for the TNF/TNFR pathway in BD. PMID- 18415773 TI - Intra-articular sodium hyaluronate 2 mL versus physiological saline 20 mL versus physiological saline 2 mL for painful knee osteoarthritis: a randomized clinical trial. AB - OBJECTIVE: Methodological constraints weaken previous evidence on intra-articular viscosupplementation and physiological saline distention for osteoarthritis. We conducted a randomized, patient- and observer-blind trial to evaluate these interventions in patients with painful knee osteoarthritis. METHODS: We centrally randomized 251 patients with knee osteoarthritis to four weekly intra-articular injections of sodium hyaluronate 2 mL (Hyalgan 10.3 mg/mL) versus physiological saline 20 mL (distention) versus physiological saline 2 mL (placebo) and followed patients for 26 weeks. Inclusion criteria were age over 59 years and daily knee pain more than 20 mm on a 100-mm visual analogue scale (VAS) without satisfactory response to analgesics. During the trial, rescue analgesic were allowed. The primary outcome was pain on movement. The secondary outcomes were pain at rest, pain during the night, Knee Injury and Osteoarthritis Outcome Score (KOOS), Osteoarthritis Research Society International (OARSI) criteria, and global assessment of the patient's condition. RESULTS: The mean age of the patients was 69.4 years; 55% were women. The effects of hyaluronate 2 mL, physiological saline 20 mL, and physiological saline 2 mL did not differ significantly in reducing knee pain, knee function, or consumption of analgesics. Using OARSI criteria, no significant differences were found. The VAS and KOOS outcomes all improved significantly over time (p<0.0005), regardless of intervention group. No adverse events were reported. CONCLUSIONS: Intra-articular hyaluronate or distention with physiological saline did not significantly reduce pain compared with physiological saline placebo in patients with osteoarthritis of the knee. PMID- 18415774 TI - Measurement characteristics of a new rapid anti-CCP2 test compared to the anti CCP2 ELISA. AB - OBJECTIVE: Anti-cyclic citrullinated peptide (CCP)2 antibody status is an important diagnostic tool in the work-up of undifferentiated arthritis (UA)/early rheumatoid arthritis (RA) but the results of the enzyme-linked immunosorbent assay (ELISA) are only available a few days after the test. The aim of this study was to assess the measurement characteristics of a rapid anti-CCP2 test compared to the usual anti-CCP2 ELISA test. METHODS: In the first phase, rapid anti-CCP2 (CCPoint) tests were performed in capillary blood obtained by finger puncture (CAP), in venous blood from a clot tube (CLOT) and in serum (SERUM) in consenting RA patients. Anti-CCP2 measured in serum using the anti-CCP2 ELISA (ELISA) was set as the gold standard (reference). In the second phase of the study, specificity versus RA was confirmed in consenting non-RA patients and healthy controls. The anti-CCP2 results were negative (no visible line or anti-CCP2<25 U/mL) or positive (visible line or anti-CCP2> or =25 U/mL). RESULTS: A total of 880 subjects (109 RA patients, 351 non-RA patients and 420 healthy controls) were enrolled in this study. For the RA patients, 5%, 15%, and 1% of CAP, CLOT and SERUM tests, respectively, were inconclusive. The sensitivity and specificity of CAP compared with ELISA were 95% (95% CI 90-100) and 95% (95% CI 89-100), respectively, and the corresponding values for SERUM were 97% (95% CI 93-100) and 98 (95% CI 94-100). The specificity for RA versus non-RA and healthy controls was 99% (95% CI 97-100) and 99% (95% CI 98-100), respectively. CONCLUSION: The CCPoint test is a fast, valid and reliable anti-CCP2 test in both capillary blood and serum but not directly in venous blood. PMID- 18415775 TI - Successful treatment with multiple intra-articular injections of infliximab in a patient with psoriatic arthritis. AB - This case report presents the clinical and radiographic course of temporomandibular joint (TMJ) involvement in a patient with severe TMJ symptoms from psoriatic arthritis (PsA) resistant to both systemic infliximab and intra articular glucocorticoid and who therefore received multiple intra-articular infliximab injections for 36 weeks. TMJ symptoms improved after the first bilateral intra-articular infliximab injections but even more so after the second injections. The considerable improvement remained for the 36 weeks studied. Bilateral computerized tomography showed no progression in radiographic changes during the treatment. No adverse reaction was observed from the intra-articular injections. PMID- 18415776 TI - Hyperpigmentation of the skin due to hydroxychloroquine. PMID- 18415777 TI - Mycobacterium chelonae infection associated with adalimumab therapy. PMID- 18415778 TI - High-resolution T2-weighted abdominal magnetic resonance imaging using respiratory triggering: impact of butylscopolamine on image quality. AB - BACKGROUND: Respiratory triggering allows the acquisition of high-resolution magnetic resonance (MR) images of the upper abdomen. However, the depiction of organs close to the gastrointestinal tract can be considerably impaired by ghosting artifacts and blurring caused by bowel peristalsis. PURPOSE: To evaluate the effect of gastrointestinal motion suppression by intramuscular butylscopolamine administration on the image quality of a respiratory-triggered T2-weighted turbo spin-echo (T2w TSE) sequence of the upper abdomen. MATERIAL AND METHODS: Images of 46 patients were retrospectively analyzed. Twenty-four patients had received intramuscular injection of 40 mg butylscopolamine immediately before MR imaging. Fourteen of the 24 patients in the butylscopolamine group underwent repeat imaging after a mean of 29 min. Quantitative analysis of the ghosting artifacts was done by measuring signal intensities in regions of interest placed in air anterior to the patient. In addition, image quality was assessed qualitatively by two radiologists by consensus. RESULTS: Spasmolytic medication with butylscopolamine reduced ghosting artifacts and significantly improved image quality of the respiratory-triggered T2w TSE sequence. The most pronounced effect of butylscopolamine administration on image quality was found for the pancreas and the left hepatic lobe. The rate of examinations with excellent or good depiction of the pancreas and the left hepatic lobe in the group without premedication and in the butylscopolamine group was 55% vs. 96% (pancreatic head), 35% vs. 88% (pancreatic body), 43% vs. 96% (pancreatic tail), and 45% vs. 83% (left hepatic lobe), respectively. Regarding the duration of the effect of intramuscular butylscopolamine, repeat imaging after a mean of 29 min did not result in a significant deterioration of image quality. CONCLUSION: Intramuscular butylscopolamine administration significantly improves image quality of respiratory-triggered T2-weighted abdominal MR imaging by persistent reduction of peristaltic artifacts. MR imaging of the liver and pancreas in particular benefits from the suppression of gastrointestinal peristalsis by butylscopolamine. PMID- 18415779 TI - High-b-value diffusion-weighted magnetic resonance imaging of pancreatic cancer and mass-forming chronic pancreatitis: preliminary results. AB - BACKGROUND: Mass-forming chronic pancreatitis may mimic a pancreatic cancer on dynamic computed tomography (CT) and magnetic resonance (MR) imaging, and preoperative differential diagnosis is often difficult. Recently, the usefulness of diffusion-weighted MR imaging (DWI) in the diagnosis of pancreatic cancer has been reported in several studies. PURPOSE: To determine whether high-b-value DWI can distinguish pancreatic cancer from benign mass-forming chronic pancreatitis. MATERIAL AND METHODS: Twenty pancreatic cancers and four cases of mass-forming chronic pancreatitis were evaluated by high-b-value DWI (b=800 s/mm(2)). The signal intensity on DWI was visually evaluated, and the isotropic apparent diffusion coefficients (ADCs) were measured. RESULTS: All twenty pancreatic cancers showed high signal intensity (18 showed very high, two showed slightly high) on DWI. None of the mass-forming chronic pancreatitis cases showed very high intensity (three showed iso to low, one showed slightly high) on DWI. The ADCs in the pancreatic cancer and mass-forming chronic pancreatitis were 1.38 +/- 0.32 x 10(-3) mm(2)/s and 1.00 +/- 0.18 x 10(-3) mm(2)/s, respectively (P < 0.05). CONCLUSION: On high-b-value DWI, most pancreatic cancers showed very high signal intensity, and may hence be distinguished from benign mass-forming chronic pancreatitis based on our preliminary results. PMID- 18415780 TI - Poland's syndrome complicated with breast cancer: mammographic, ultrasonographic, and computed tomographic findings. AB - Poland's syndrome is a rare congenital anomaly characterized by unilateral chest wall hypoplasia and ipsilateral hand abnormalities. It has been reported in association with various malignancies and other developmental defects. We report here the case of a 58-year-old woman with Poland's syndrome who developed breast cancer in the ipsilateral normal breast. A review of the literature reveals that two studies of breast carcinoma associated with Poland's syndrome have been reported, but this paper is the first example of a carcinoma occurring in an otherwise normal breast associated with Poland's syndrome. PMID- 18415781 TI - Prophylactic implantation of inferior vena cava filter during endovascular therapies for deep venous thrombosis of the lower extremity: is it necessary? AB - BACKGROUND: There is strong controversy as to whether to use filters temporarily in the inferior vena cava (IVC) during endovascular therapies for deep venous thrombosis (DVT) in the lower extremity. PURPOSE: To evaluate the necessity of implantation of a retrievable inferior vena cava filter during treatment of DVT in the lower extremity using various endovascular techniques. MATERIAL AND METHODS: Studied were all 37 consecutive sessions of endovascular treatments performed in 34 patients with symptomatic lower-limb DVT between April 2001 and October 2006. In all sessions, a Gunther tulip retrievable vena cava filter (GTF) was implanted during the procedures. The degree of trapped thrombus in the filter was evaluated by repeated venocavography. RESULTS: Implantation of a total of 66 GTFs was successfully performed in 37 sessions. Worsening of or new formation of pulmonary embolism was avoided in all patients. In 20 (54.1%) of the 37 sessions, a trapped thrombus in the filter was observed. In 14, the trapped thrombus was large, filling more than half the height of the filter. CONCLUSION: Temporary implantation of filters is effective in preventing pulmonary emboli during endovascular treatment of DVT in the lower extremities. PMID- 18415782 TI - Patient number calculations. PMID- 18415783 TI - Clinical value of 16-slice multidetector computed tomography in symptomatic patients with suspected coronary artery disease. AB - BACKGROUND: Multidetector computed tomography angiography has emerged as a rapidly developing method for the noninvasive detection of coronary artery disease. PURPOSE: To investigate the diagnostic accuracy and limitations of multidetector computed tomography (MDCT) in the detection of significantly (> 50%) obstructive coronary artery disease (CAD) using a scanner equipped with 16 x 0.625-mm collimation. MATERIAL AND METHODS: MDCT angiography was performed in 153 patients (99 male, 54 female; mean age 55 +/- 10 years) with suspected CAD and scheduled for conventional coronary angiography (CCA). Image quality was assessed in terms of artifacts and segment visibility, and the assessable segments were screened for the presence of significant stenoses (> 50% lumen diameter reduction). The diagnostic performance of MDCT for the detection of significant stenosis was compared with the results of CCA. RESULTS: In all 153 patients, MDCT was carried out without complications. A total of 1989 coronary artery segments were evaluated. After exclusion of 394 non-evaluable segments (19.8%), 1595 segments (80.2%) were included in the analysis. The most frequent causes of poorly assessable segments were motion artifact (36%) and severe calcification (23%). Considering only the segments judged to be evaluable, the sensitivity, specificity, and positive and negative predictive values of 16-slice MDCT were 85%, 97%, 79%, and 98%, respectively. Including all segments in the analysis (evaluable and non-evaluable), the sensitivity was 74%, specificity 96%, positive predictive value 73%, and negative predictive value 97%. CONCLUSION: When all coronary artery segments are included, 16-slice MDCT has moderate sensitivity and very high specificity and negative predictive value in assessing coronary artery stenoses. High specificity and negative predictive value indicate that 16-slice MDCT may be a useful tool in reliably ruling out significant lesions in patients with a low pretest probability. PMID- 18415784 TI - Iodixanol compared to iohexol for contrast procedures: a case-matched retrospective cohort study. AB - BACKGROUND: Previous studies that have attempted to evaluate the effectiveness of an iso-osmolar contrast medium (IOCM) iodixanol compared to a low-osmolar contrast medium (LOCM) for contrast procedures show variable results. PURPOSE: To evaluate the nephrotoxicity of the IOCM iodixanol compared to the LOCM iohexol. MATERIAL AND METHODS: We performed a retrospective cohort study from April 2004 to March 2006. All contrast procedures with a pre- and post-exposure creatinine value were evaluated for inclusion. Contrast nephropathy (CN) was defined as post exposure creatinine elevation of > or = 25% or > 0.5 mg/dl within 7 days of contrast exposure. Cases of iodixanol exposure were matched to control cases of iohexol exposure (1:1) based on age, sex, presence of diabetes, pre-exposure creatinine value, and type of imaging study performed. We matched 397 cases of iodixanol (IOCM) exposure to 397 cases of iohexol (LOCM) exposure. Cases of iodixanol which could not be matched to controls were not included in the analysis. RESULTS: After adjustment for prior creatinine, medications, contrast iodine load, prior exposure to contrast material, heart failure, and hypertension, use of iodixanol did not significantly alter rates of CN compared to iohexol (OR 0.92, 95% CI 0.57-1.46; P = 0.71) or mortality (RR 0.79, 95% CI 0.59-1.06; P = 0.12). CONCLUSION: The use of the IOCM iodixanol was not associated with statistically significant protection against contrast nephropathy or all-cause mortality compared to a matched cohort of patients receiving the LOCM iohexol. PMID- 18415785 TI - Assessing doses to interventional radiologists using a personal dosimeter worn over a protective apron. AB - BACKGROUND: Interventional radiologists receive significant radiation doses, and it is important to have simple methods for routine monitoring of their exposure. PURPOSE: To evaluate the usefulness of a dosimeter worn outside the protective apron for assessments of dose to interventional radiologists. MATERIAL AND METHODS: Assessments of effective dose versus dose to dosimeters worn outside the protective apron were achieved by phantom measurements. Doses outside and under the apron were assessed by phantom measurements and measurements on eight radiologists wearing two routine dosimeters for a 2-month period during ordinary working conditions. Finger doses for the same radiologists were recorded using thermoluminescent dosimeters (TLD; DXT-RAD Extremity dosimeters). RESULTS: Typical values for the ratio between effective dose and dosimeter dose were found to be about 0.02 when the radiologist used a thyroid shield and about 0.03 without. The ratio between the dose to the dosimeter under and outside a protective apron was found to be less than 0.04. There was very good correlation between finger dose and dosimeter dose. CONCLUSION: A personal dosimeter worn outside a protective apron is a good screening device for dose to the eyes and fingers as well as for effective dose, even though the effective dose is grossly overestimated. Relatively high dose to the fingers and eyes remains undetected by a dosimeter worn under the apron. PMID- 18415786 TI - Diffusion-weighted imaging for predicting new compression fractures following percutaneous vertebroplasty. AB - BACKGROUND: Percutaneous vertebroplasty (PVP) is a technique that structurally stabilizes a fractured vertebral body. However, some patients return to the hospital due to recurrent back pain following PVP, and such pain is sometimes caused by new compression fractures. PURPOSE: To investigate whether the apparent diffusion coefficient (ADC) of adjacent vertebral bodies as assessed by diffusion weighted imaging before PVP could predict the onset of new compression fractures following PVP. MATERIAL AND METHODS: 25 patients with osteoporotic compression fractures who underwent PVP were enrolled in this study. ADC was measured for 49 vertebral bodies immediately above and below each vertebral body injected with bone cement before and after PVP. By measuring ADC for each adjacent vertebral body, ADC was compared between vertebral bodies with a new compression fracture within 1 month and those without new compression fractures. In addition, the mean ADC of adjacent vertebral bodies per patient was calculated. RESULTS: Mean preoperative ADC for the six adjacent vertebral bodies with new compression fractures was 0.55 x 10(-3) mm(2)/s (range 0.36-1.01 x 10(-3) mm(2)/s), and for the 43 adjacent vertebral bodies without new compression fractures 0.20 x 10(-3) mm(2)/s (range 0-0.98 x 10(-3) mm(2)/s) (P < 0.001). Mean preoperative ADC for the six patients with new compression fractures was 0.55 x 10(-3) mm(2)/s (range 0.21-1.01 x 10(-3) mm(2)/s), and that for the 19 patients without new compression fractures 0.17 x 10(-3) mm(2)/s (range 0.01-0.43 x 10(-3) mm(2)/s) (P < 0.001). CONCLUSION: The ADC of adjacent vertebral bodies as assessed by diffusion weighted imaging before PVP might be one of the predictors for new compression fractures following PVP. PMID- 18415787 TI - Comparative study of fluorodeoxyglucose positron emission tomography and magnetic resonance imaging for the detection of spinal bone marrow infiltration in untreated patients with multiple myeloma. AB - BACKGROUND: The presence and extent of osteolytic bone lesions in untreated patients with multiple myeloma are important factors in the staging of the disease, and the extent of bone lesions in multiple myeloma cases significantly influences decisions regarding therapy. Recently, fluorodeoxyglucose positron emission tomography (FDG-PET) and magnetic resonance imaging (MRI) have been used to detect bone marrow involvement in patients with multiple myeloma. PURPOSE: To compare the efficacy of FDG-PET and MRI for the detection of bone marrow infiltration into the spine in untreated patients with multiple myeloma. MATERIAL AND METHODS: Twenty-two patients with multiple myeloma underwent both FDG-PET and spine MRI. The examined spinal regions by MRI included 21 thoracic and lumbar spines, one lumbar spine, and 12 cervical spines. The following imaging sequences were performed: T1-weighted spin-echo MRI with and without fat suppression, and T2-weighted spin-echo MRI in the sagittal plane. In the patients with bone marrow abnormalities, an additional contrast-enhanced T1-weighted spin-echo MR image and a fat-suppressed T1-weighted spin-echo MR image were obtained. Patients were divided into three groups on the basis of the criteria defined by Durie and Salmon: stage I (n=9), stage II (n=3), and stage III (n=10). The number and location of lesions detected in both FGD-PET and MRI were recorded, and the lesions were compared using the McNemar test. Bone marrow biopsy results, the patient's clinical examinations, and other imaging findings (MRI, FDG-PET, etc.) were used as references. RESULTS: In stages I and II (37 lesions in 12 patients), FDG-PET and MRI detected lesions in 78% (29 of 37 lesions) and 86% (32 of 37 lesions), respectively. However, the difference between the abilities of FDG-PET and MRI to detect lesions was not statistically significant (P=0.317). In stage III (101 lesions in 10 patients), FDG-PET and MRI detected lesions in 80% (81 of 101 lesions) and 92% (93 of 101 lesions), respectively. The difference between the abilities of FDG-PET and MRI to detect lesions was statistically significant (P=0.038). CONCLUSION: MRI is superior to FDG-PET in detecting bone marrow involvement in the spine of patients with advanced multiple myeloma. PMID- 18415788 TI - The prevalence of cam-type deformity of the hip joint: a survey of 4151 subjects of the Copenhagen Osteoarthritis Study. AB - BACKGROUND: Cam deformity is a preosteoarthritic malformation causing premature hip-joint degeneration. While the pathogenetic pathway from deformity to osteoarthrosis (OA) has been well established, almost nothing is known of the malformation's epidemiology. PURPOSE: To determine the distribution of cam deformity in a large, unselected cohort from standardized anteroposterior (AP) pelvic radiographs. MATERIAL AND METHODS: The distribution of cam deformity was assessed in 3202 (1184 male, 2018 female) standardized AP pelvic radiographs using the triangular index (TI) and the alpha angle. The relationships between cam malformation and self-reported hip pain were evaluated, and the relative importance of known risk factors for cam malformation estimated. RESULTS: We found a pronounced sex-related difference in cam-deformity distribution. The overall prevalence of cam deformity was approximately 17% in men and 4% in women. The distribution of cam deformity was unaltered in subjects with normal joint space width or other features of hip-joint degeneration. We found no significant association with self-reported hip pain, nor did we find any relative importance of possible risk factors for hip deformity, such as body-mass index (BMI), occupational exposure to heavy workloads, or concomitant acetabular dysplasia. CONCLUSION: The results lend support to the thesis that cam deformity represents a silent slipped capital epiphysis, predominantly in men, and that it is a far from uncommon deformity in subjects with no apparent evidence of hip-joint osteoarthritis. PMID- 18415789 TI - Assessment of wall shear stress in the common carotid artery of healthy subjects using 3.0-tesla magnetic resonance. AB - BACKGROUND: Wall shear stress (WSS) has been proven to play a critical role in the formation and development of atherosclerotic plaques. Measurement of WSS in vivo is significant for the clinical assessment of atherosclerosis. PURPOSE: To assess the magnitude and distribution of local WSS in the common carotid artery (CCA) in vivo using 3.0T magnetic resonance (MR). MATERIAL AND METHODS: The common carotid artery of eight healthy volunteers was studied using a cine phase contrast MR sequence. A three-dimensional paraboloid model was applied to fit the velocity profiles, and the WSS values were calculated. The cross-sectional area, average flow velocity, maximum velocity, and flow rate were also obtained. RESULTS: Mean WSS was 0.850 +/- 0.195 (range 0.132-3.464) N/m(2) for the common carotid arteries; the spatial and temporal distribution and change of WSS were displayed. During a cardiac cycle, the mean velocity was 22.8 +/- 3.5 (16.9-28.3) cm/s, blood flow rate 8.03+/-1.45 (5.73-10.72) ml/s, and luminal vessel area 34.94+/-7.06 (24.25-49.01) mm(2). CONCLUSION: Local WSS values in CCAs can be measured using 3.0T MR imaging combined with image-processing techniques. Intersubject variations were found in the distribution and magnitude of wall shear stress as well as in the flow profile pattern in CCAs, which may be caused by different vessel morphologies. PMID- 18415790 TI - Diagnostic accuracy and interobserver variability of pulsed arterial spin labeling for glioma grading. AB - BACKGROUND: Although pulsed arterial spin labeling (PASL) enables the reliable qualitative grading of brain tumors, its use in quantification for glioma grading may be hampered by the limited interobserver variability associated with low spatial resolution. PURPOSE: To assess the interobserver variability and diagnostic accuracy of the relative tumor perfusion signal intensity (rTPS) calculated using PASL in glioma grading. MATERIAL AND METHODS: Fifty-eight patients with 61 cerebral astrocytomas underwent conventional MR imaging and PASL. Receiver operating characteristic analyses were used to determine the optimum thresholds for tumor grading. The sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) for identifying high grade gliomas were also calculated. Cohen's kappa statistic was used to determine the levels of interobserver variability in the quantitative analysis of PASL. RESULTS: The sensitivity, specificity, PPV, and NPV for determining a high-grade glioma with conventional MR imaging were 77.1, 73.1, 79.4, and 70.4%, respectively. A threshold value of 1.28 for rTPS provided a sensitivity, specificity, PPV, and NPV of 82.9, 96.2, 96.7, and 80.6%, respectively. There was a statistically significant difference in the rTPS between low- and high-grade astrocytomas (1.14 vs. 1.47, P<0.05). In the interobserver variability analysis, substantial agreement was obtained for the quantitative rTPS measurement from PASL (kappa =0.72). CONCLUSION: Quantitative perfusion measurement with PASL can improve the diagnostic accuracy of preoperative glioma grading, as compared to the application of conventional imaging alone. However, the interobserver variability for quantification is substantial. PMID- 18415791 TI - Tuberculosis of the parotid gland: computed tomographic findings. AB - BACKGROUND: Tuberculosis (TB) of the parotid gland is rare, even in endemic regions. Approximately 100 cases have been reported in the English literature. The computed tomographic (CT) features, however, have seldom been studied. PURPOSE: To determine the diagnostic CT features of tuberculosis of the parotid gland. MATERIAL AND METHODS: CT studies of four histologically proven cases of tuberculosis of the parotid gland were retrospectively reviewed. RESULTS: A total of 15 enlarged lymph nodes were found in the superficial lobes of the parotid glands. The nodes were arranged linearly within the gland. Enhancement patterns included homogeneous enhancement (9/15, 60%), homogeneous enhancement with eccentric microcysts (3/15, 20%), and thick-walled rim enhancement with central lucency (3/15, 20%). Thickened adjacent fascial plane and platysma were seen in two patients. Ipsilateral cervical lymphadenopathy was seen in all patients. CONCLUSION: In patients presenting with unilateral parotid nodules, TB should be considered when linearly arranged enhancing nodules are demonstrated in the superficial lobes of the glands on CT scan. PMID- 18415792 TI - Magnetic resonance imaging of pilocytic astrocytomas: usefulness of the minimum apparent diffusion coefficient (ADC) value for differentiation from high-grade gliomas. AB - BACKGROUND: On contrast-enhanced magnetic resonance (MR) images, pilocytic astrocytomas (PAs) are usually well-enhanced tumors that may mimic high-grade gliomas (HGGs). On the other hand, it has been suggested that areas exhibiting minimum apparent diffusion coefficient (ADC) values reflect the sites of highest cellularity within heterogeneous tumors. PURPOSE: To test the hypothesis that the cellularity of PAs is significantly different to the cellularity of HGGs, which should result in significant differences in minimum ADC values. MATERIAL AND METHODS: Between 1999 and 2005, 15 patients (nine males, six females) with histopathologically confirmed PAs underwent pretreatment MR examination including diffusion-weighted (DW) imaging. We reviewed their MR findings with respect to the size, location, morphology, contrast enhancement, and minimum ADC value of the tumors. The minimum ADC values of the 15 PAs were compared with those of 104 HGGs diagnosed during the same period. RESULTS: The diameter of the 15 PAs ranged from 11 to 60 mm (mean 36 mm); all were located around the ventricles, and all contained enhancing components. All except two small (11 and 14 mm) PAs contained cystic components. The minimum ADC values were significantly higher in PAs (median 1.688, range 1.375-1.897 x 10(-3) mm(2)/s) than HGGs (0.997, 0.543-2.024 x 10(-3) mm(2)/s) (P < 0.0001), although there was substantial overlap. Among the tumors with enhancing components, all but one PA were differentiated from the 76 HGGs with enhancing components (0.922, 0.543-1.462 x 10(-3) mm(2)/s) when the minimum ADC cutoff value was set at 1.5 x 10(-3) mm(2)/s. CONCLUSION: The minimum ADC value may be helpful for the differentiation between PAs and HGGs. A tumor with enhancing components should be PA instead of HGG when the minimum ADC value is higher than 1.5 x 10(-3) mm(2)/s. PMID- 18415793 TI - Clinical evaluation of a dual-side readout technique computed radiography system in chest radiography of premature neonates. AB - BACKGROUND: Recently, the dual-side readout technique has been introduced in computed radiography, leading to an increase in detective quantum efficiency (DQE) compared with the single-side readout technique. PURPOSE: To evaluate if the increase in DQE with the dual-side readout technique results in a higher clinical image quality in chest radiography of premature neonates at no increase in radiation dose. MATERIAL AND METHODS: Twenty-four chest radiographs of premature neonates were collected from both a single-side readout technique system and a double-side readout technique system. The images were processed in the same image-processing station in order for the comparison to be only dependent on the difference in readout technique. Five radiologists rated the fulfillment of four image quality criteria, which were based on important anatomical landmarks. The given ratings were analyzed using visual grading characteristics (VGC) analysis. RESULTS: The VGC analysis showed that the reproduction of the carina with the main bronchi and the thoracic vertebrae behind the heart was better with the dual-side readout technique, whereas no significant difference for the reproduction of the central vessels or the peripheral vessels could be observed. CONCLUSIONS: The results indicate that the higher DQE of the dual-side readout technique leads to higher clinical image quality in chest radiography of premature neonates at no increase in radiation dose. PMID- 18415795 TI - Pediatric multidetector computed tomography using tube current modulation and a patient image gallery. AB - BACKGROUND: Dose reduction is crucial in pediatric multidetector computed tomography (MDCT). PURPOSE: To perform pediatric 16-slice MDCT using tube current modulations and to adjust prospectively the tube current using a patient image gallery (IG) providing simulated dose-reduced protocols; and to evaluate and compare the image quality of the IG and the clinical MDCT. MATERIAL AND METHODS: 30 examinations (thorax, n = 15; abdomen, n = 8; pelvis, n = 7) in 20 patients (nine male, age 8.05 +/- 7.33 years, weight 29.8 +/- 24.02 kg) were performed according to an IG on a 16-slice MDCT with additional use of online tube current modulation (CARE Dose). Three radiologists visually assessed image quality from the IG and actual 16-slice MDCT scans. For objective analysis, image noise was determined. RESULTS: Statistical analysis showed moderate concordance in objective (K = 0.68-0.78) and subjective (K = 0.33-0.64) image assessment between the IG and clinical 16-slice MDCT scans. Depending on the weight group and clinical question, no or only minor dose reductions in the chest, but moderate to considerable reductions in the abdominal/pelvic 16-slice MDCT scans compared to previously used pediatric protocols were achieved. Extra dose reduction was achieved due to additional use of CARE Dose depending on age group and scan region (mean 8.6-23.9%). CONCLUSION: The IG enabled us to prospectively reduce the tube current and adapt the required image quality to the clinical question. Additional dose reduction was achieved with application of CARE Dose; nevertheless, the images are comparable to the simulated images of the IG. PMID- 18415796 TI - Critical review of Clara cell protein: sound science? PMID- 18415798 TI - Plasma biomarker profiling in the detection of growth promoter use in calves. AB - The detection of illicit growth promoter use during meat production within the European Union is reliant on residue testing which is a limiting factor on the number of animals which can be tested and consequently compromises the efficacy of testing procedures. The present study examined a novel detection strategy based on the profiling of plasma component concentrations in response to growth promoter administrations. Calves subjected to nortestosterone decanoate, 17beta oestradiol benzoate and dexamethasone were found to have altered urea, aminoterminal propeptide of type III procollagen and sex hormone binding globulin profiles in response to treatments. These findings demonstrate the potential of using the identification of perturbed profiles within a panel of biomarkers which cover a spectrum of biological activity to reveal growth promoter abuse. PMID- 18415799 TI - ICAM-1 expression in vaginal cells as a potential biomarker for inflammatory response. AB - This study aimed to elucidate the mechanisms that may lead to an efficient strategy to induce a suitable host response of the vaginal mucosa upon exposure to intravaginally delivered exogenous compounds. It was hypothesized that the upregulation of intercellular adhesion molecule (ICAM)-1 gene expression may reflect the inflammatory response evoked by exogenous compounds. Major emphasis was placed on ethylenediamine tetraacetic acid (EDTA) which was added as a synergistic agent to conventional spermicidal agents or anti-HIV drugs. The levels of ICAM-1 mRNA were examined as a surrogate marker for inflammatory response in human vaginal epithelial cells upon exposure to EDTA or interleukin (IL)-1beta (i.e. positive control, 25 mM). The effects of estrogen on EDTA induced ICAM-1 expression were also evaluated for the estrogen involvement in the inflammatory process of the vaginal mucosa. ICAM-1 expression in human vaginal cells (VK2/E6E7 cells) increased as EDTA concentration added to human vaginal cell lines increased. The effects of estrogen on EDTA-induced ICAM-1 expression in human vaginal epithelial cells were estrogen-concentration dependent; estrogen at lower concentrations (approximately 1-10 nM) did not affect ICAM-1 expression, whereas estrogen at higher concentrations (approximately 100 nM-1 microM) attenuated ICAM-1 expression. The influence of estrogen in ICAM-1 expression suggests the beneficial effects of estrogen on the regulation of vaginal homeostasis. Identification and quantification of specific surrogate markers for the inflammatory response evoked by exogenous compounds and their regulation by estrogen will lead to an efficient strategy against sexually transmitted diseases including AIDS. PMID- 18415800 TI - Plasma concentrations of matrix metalloproteinase-2, tissue inhibitor of metalloproteinase-1 and osteopontin reflect severity of heart failure in DOCA salt hypertensive rat. AB - The matrix metalloproteinases (MMPs) and their endogenous inhibitors, the tissue inhibitors of metalloproteinases (TIMPs) play a key role in extracellular matrix maintenance and are altered in the failing heart, both in experimental models and in chronic end-stage heart failure in humans. As the common diagnostic markers of heart failure, atrial natriuretic peptide (ANP) and B-type natriuretic peptide (BNP) primarily reflect increased pressure loading, determination of soluble, heart-derived MMPs and TIMPs in plasma, as well as the determination of the emerging fibrosis marker osteopontin (OPN) might be valuable tools for detecting heart fibrosis. In this study the effect of spironolactone treatment on plasma MMP-2, TIMP-1 and OPN levels was assessed in a heart failure animal model. Unilaterally nephrectomized Sprague Dawley rats received subcutaneous injection of 100 mg deoxycorticosterone acetate (DOCA) once a week and 1% (w/v) NaCl in drinking water. Blood pressure was monitored weekly and blood samples were collected after 1, 2 and 4 weeks. After 6 weeks, left ventricular contractility (LVC) and heart weight-to-body weight ratio (HW/BW) were assessed. DOCA treatment increased plasma MMP-2, TIMP-1 and OPN concentrations. Alterations of plasma marker levels were correlated with changes of HW/BW and paralleled impaired LVC. Furthermore, beneficial effects of spironolactone treatment were observed. In DOCA-salt hypertensive rats, plasma concentrations of MMP-2, TIMP-1 and OPN reflected heart failure associated with haemodynamic, functional and morphological changes. Based on these findings, it appears reasonable to use plasma markers of fibrosis to monitor the development of heart failure. PMID- 18415801 TI - Combined effect of GSTM1, GSTT1 and GSTP1 polymorphisms on histological subtypes of lung cancer. AB - Genetic polymorphisms are natural genetic variations in the gene sequence that occur at a frequency of >1% in the population. This genetic variability (polymorphisms) can be a factor in cancer risk. The functional polymorphisms in GST genes play an important role in susceptibility to lung cancer. In our previous study, we reported that the combination of certain genotypes of GSTM1, GSTT1 and CYP1A1 is associated with lung cancer. The study has been extended to investigate the potential role of polymorphism in GSTP1 alone or in combination with the status of GSTM1 and GSTT1 genes in the likelihood of development of lung cancer. A total of 302 subjects (151 cases and 151 controls) were evaluated. Using a case-control design, individuals were genotyped for GSTs using multiplex polymerase chain reaction and restriction fragment length polymorphism techniques. The data obtained were analyzed using multiple logistic regression. The combined 'at risk' genotypes of GSTM1 null and GSTT1 null in comparison with 'wild-type' genotypes seems to be associated with a greater risk of lung cancer, but the results are not significant (odds ratio (OR) 2.0, 95% confidence interval (CI) 0.68-5.96) and for squamous cell carcinoma (SqCC) it was 1.6-fold (OR 1.6, 95% CI 0.49-5.68). In summary, our case-control study of lung cancer revealed that the effect of these polymorphisms is not very marked for different genotypic combinations of GSTP1, GSTM1 and GSTT1 in the context of developing lung cancer in a north Indian population. However, the increased risk was limited to SqCC, and was not found for other histological subtypes. Further analyses on this topic are needed. PMID- 18415802 TI - Ethnic groups and high sensitivity C-reactive protein in Israel. AB - High-sensitivity C-reactive protein (hs-CRP) is a biomarker that correlates with atherothrombotic risk and outcome. hs-CRP is influenced by various modifiable and non-modifiable factors. We studied the relationship between ethnic background and hs-CRP level, among the Jewish population in Israel. A total of 3659 men and 2180 women were divided into two ethnic groups (Ashkenazi and Sephardic Jews), based on the knowledge of Jewish immigration patterns throughout the centuries. Mean hs CRP levels were calculated for each group and were adjusted for various factors known to influence hs-CRP. Sephardic Jews were found to have higher adjusted mean hs-CRP levels (2.0 mg l(-1) for men and 3.9 mg l(-1) for women) compared with Ashkenazi Jews (1.5 mg l(-1) for men and 2.9 mg l(-1) for women). Ethnic background emerged as an independent significant predictor of hs-CRP levels. We demonstrated that ethnicity is an important factor when considering hs-CRP as a marker of atherothrombotic risk. PMID- 18415803 TI - Variant CD44 expression by human fibroblasts. AB - CD44 is a cell surface glycoprotein with roles in tumour invasion and metastasis. CD44 is variably spliced from ten variant exons and mis-splicing is a biomarker for detection of colon, urothelial and other carcinomas. Fibroblasts are normally considered to lack variant exons and thus should not generate false-positive signals. Transcription of variant exons by fibroblasts was investigated by exon specific reverse transcription-polymerase chain reaction (RT-PCR) for variant exons v2-v10 using normal primary fibroblasts, immortalized and experimentally transformed fibroblasts. Flow cytometry, immunocytochemistry and Western blotting were used to determine expression. All types of fibroblasts, including normal primary culture fibroblasts, transcribed low levels of variant exon mRNA. Expression could not be detected by blotting or immunocytochemistry but flow cytometry revealed minor expression of some exons by all three types of cultured fibroblast. Fibroblasts do transcribe and express small amounts of variant exon CD44. This may need to be considered when using exon splicing as a biomarker for malignancy in clinical samples containing connective tissue. PMID- 18415804 TI - Evaluation of H2O2 and pH in exhaled breath condensate samples: methodical and physiological aspects. AB - This veterinary study is aimed at further standardization of H(2)O(2) and pH measurements in exhaled breath condensate (EBC). Data obtained in the study provide valuable information for many mammalian species including humans, and may help to avoid general pitfalls in interpretation of EBC data. EBC was sampled via the 'ECoScreen' in healthy calves (body weight 63-98 kg). Serum samples and condensates of ambient (indoor) air were collected in parallel. In the study on H(2)O(2), concentrations of H(2)O(2) in EBC, blood and ambient air were determined with the biosensor system 'ECoCheck'. In EBC, the concentration of H(2)O(2) was found to be dependent on food intake and increased significantly in the course of the day. Physiologically, lowest H(2)O(2) concentrations at 06:00 varied within the range 138-624 nmol l(-1) EBC or 0.10-0.94 nmol per 100 l exhaled breath and individual concentrations were significantly different indicating a remarkable intersubject variability. Highly reproducible results were seen within each subject (three different days within 4 weeks). No correlation existed between H(2)O(2) concentrations in EBC and blood, and EBC H(2)O(2) was not influenced by variables of spontaneous breathing. Further results confirmed that standardization of H(2)O(2) measurements in EBC requires (1) the re-calculation of the concentration exhaled per 100 l exhaled breath (because the analyzed concentration in the liquid condensate underlies multiple methodological sources of variability given by the collection process), and (2) subtracting the concentration of inspired indoor H(2)O(2). In the study on pH use of the ISFET electrode (Sentron, the Netherlands) and a blood gas analyzer ABL 550 (Radiometer, Denmark) led to comparable results for EBC-pH (r=0.89, R(2)=79.3%, p or = 1 post-randomization rehabilitation session, LVRS patients, patients with an FEV(1) > or = 20% predicted, and higher education were significantly more likely to complete rehabilitation. Patients with depressive and anxiety symptoms, and those who live > 36 miles compared to < 6 miles away were less likely to be adherent. Patients who underwent LVRS completed more exercise sessions than those in the medical group and were more likely to be adherent with post-randomization rehabilitation. A better understanding of patient factors such as socioeconomic status, depression, anxiety and transportation issues may improve adherence to pulmonary rehabilitation. PMID- 18415821 TI - Outpatient evaluation of vision and ocular motricity in 123 children with cerebral palsy. AB - PURPOSE: Children with cerebral palsy (CP) may present varied ophthalmological problems. Paradoxically, however, investigation of visual function and possible related disorders is not routinely conducted in this population during medical consultations. METHOD: This paper proposes a simple, practical guide for assessing vision and ocular motricity in children with CP. One-hundred and twenty three patients (mean age: 8.4 years +/- 2.3) from the Sarah Network of Rehabilitation Hospitals (Brasilia, Rio de Janeiro and Sao Luis units) were studied. Various parameters were investigated (smooth pursuit movements, strabismus, visual acuity, visual field, visuospatial neglect and nystagmus) using the following assessment tools: adapted smooth pursuit test for ocular motricity; Snellen test for visual acuity; confrontation method for visual field; cancellation test (star test) for visual neglect; and cover test for strabismus. Nystagmus was assessed based on patient observation. RESULTS: Eighty-one children completed all the evaluations. Among these, 38 (47%) had no visual problems (hemiplegia 60.5%; diplegia 41.4%; triplegia 12.5%; mixed tetraplegic 33.3%); 23 had difficulties in one of the tested items; and 20 had problems in two or more areas of vision. CONCLUSIONS: Visual and oculomotor tests are of significant importance in children with CP and provide relevant information for creating a rehabilitation programme aimed at the individual as a whole. PMID- 18415822 TI - Response of older patients with IgE-mediated asthma to omalizumab: a pooled analysis. AB - Asthma in older adults is under-recognized and possibly associated with allergic triggers. We conducted a pooled analysis of omalizumab double-blind, placebo controlled trials to evaluate efficacy in older adults. Data for the total study population and subjects aged > or = 50 years with moderate-severe allergic asthma were examined. We used Poisson regression to analyze the number of asthma exacerbations and logistic regression to evaluate treatment effectiveness. Symptom scores and total rescue medication puffs were evaluated by analysis of covariance. Omalizumab reduced the risk of clinically significant asthma exacerbations, led to a significantly greater response in patient/investigator reported global effectiveness, improved asthma symptom scores, and reduced rescue medication use in adults > or = 50 years with moderate-severe allergic asthma. PMID- 18415810 TI - Exercise testing in severe emphysema: association with quality of life and lung function. AB - Six-minute walk testing (6MWT) and cardiopulmonary exercise testing (CPX) are used to evaluate impairment in emphysema. However, the extent of impairment in these tests as well as the correlation of these tests with each other and lung function in advanced emphysema is not well characterized. During screening for the National Emphysema Treatment Trial, maximum ergometer CPX and 6MWT were performed in 1,218 individuals with severe COPD with an average FEV(1) of 26.9 +/ 7.1 % predicted. Predicted values for 6MWT and CPX were calculated from reference equations. Correlation coefficients and multivariable regression models were used to determine the association between lung function, quality of life (QOL) scores, and exercise measures. The two forms of exercise testing were correlated with each other (r = 0.57, p < 0.0001). However, the impairment of performance on CPX was greater than on the 6MWT (27.6 +/- 16.8 vs. 67.9 +/- 18.9 % predicted). Both exercise tests had similar correlation with measures of QOL, but maximum exercise capacity was better correlated with lung function measures than 6-minute walk distance. After adjustment, 6MWD had a slightly greater association with total SGRQ score than maximal exercise (effect size 0.37 +/- 0.04 vs. 0.25 +/- 0.03 %predicted/unit). Despite advanced emphysema, patients are able to maintain 6MWD to a greater degree than maximum exercise capacity. Moreover, the 6MWT may be a better test of functional capacity given its greater association with QOL measures whereas CPX is a better test of physiologic impairment. PMID- 18415824 TI - The value of exhaled nitric oxide in predicting bronchial hyperresponsiveness in children. AB - Reduced attention span and motor skills in children limit the practicability of bronchial provocation tests. To assess exhaled nitric oxide (FeNO) as a surrogate for bronchial hyperresponsiveness (BHR) in children with possible reactive airway disease, FeNO was measured using the single-breath method in 169 successive outpatients 11 +/- 5 years of age before lung function testing and subsequent bronchial provocation by exercise (n = 165) and methacholine (n = 134). Baseline forced expiratory volume in 1 second (FEV(1)) less than 80% of predicted and/or BHR were seen in 59%. FeNO correlated weakly with PD(20) to methacholine (r = 0.24, p < 0.05), but not with the change in FEV(1) due to exercise-induced bronchoconstriction (EIB) (r = 0.1, p > 0.05). The negative predictive value of FeNO less than 10 ppb for EIB was 94%, but overall accuracy for predicting BHR was low. Measurement of FeNO is not a substitute for bronchial provocation in children. PMID- 18415825 TI - Happy Air: a school-based educational program to maximize detection of asthma in children. AB - OBJECTIVE: To investigate whether an active partnership among school, parents, and pediatricians allows early identification and treatment of asthmatic children. METHODS: An asthma educational program (Happy Air), based on a strong family-physician-school interrelationship, was performed in six primary schools (2,765 children) before administering a screening questionnaire to the parents. RESULTS: A high response rate (96%) demonstrated 2,649 responders available for the asthma screening: 135 children (5%) received a diagnosis of asthma, of which 37 (27%) were recognized de novo. CONCLUSION: The active participation of school and parents is the determining factor for the success of an asthma screening program. PMID- 18415823 TI - Sleep quality in asthma: results of a large prospective clinical trial. AB - RATIONALE: Previous studies have suggested that asthmatics have an increased incidence of sleep disturbances. However, these studies have been limited by reliance on population surveys or small numbers of participants. OBJECTIVES: We sought to measure sleep quality and daytime sleepiness in a cohort of symptomatic asthmatics and to measure the effects of improved asthma control on sleep quality. METHODS: Data were collected in sub-study of a large multi-center randomized double-masked controlled trial of mild-moderate asthmatics evaluating the effect of low-dose theophylline on asthma control in comparison to montelukast and placebo. Each participant was administered sleep symptom questionnaires at randomization and at the final visit (6 months after randomization). These included the Pittsburgh Sleep Quality Questionnaire (PSQI) and the Epworth Sleepiness Scale (ESS). MEASUREMENTS AND MAIN RESULTS: Data were available for 487 participants. Baseline mean values were: age 40 +/- 15 years, 74% female, forced expiratory volume in 1 second (FEV(1)) 79 +16 percent predicted, Juniper Asthma Control Questionnaire (ACQ) score 2.35 +/- 0.63, PSQI 7.8 +/-4, and ESS 8.5 +/-4.9. There were no significant differences in the PSQI or ESS between the three treatment groups. Significant correlations were found at baseline between the global PSQI score and ACQ and quality of life and marginally with lung function. Significant correlation existed between improvements in PSQI and ESS with improved asthma control and quality of life. CONCLUSIONS: Sleep disturbances are common in asthmatics and are associated with asthma control and quality of life. Clinicians caring for asthmatics may need to complete a more detailed sleep history in patients with poorly controlled asthma. In addition, low-dose theophylline does not seem to impair sleep quality in asthmatics. PMID- 18415826 TI - Effect of budesonide and formoterol on IL-6 and IL-8 release from primary bronchial epithelial cells. AB - The aim of this study was to explore possible interactions between a glucocorticosteroid (budesonide) and a long-acting beta(2)-agonist (formoterol) on pro-inflammatory cytokine release. Primary bronchial epithelial cells (PBEC) were stimulated with organic dust and incubated with budesonide, formoterol, and the combination of both drugs. Interleukin (IL)-6 and -8 in the supernatant was assessed by enzyme-linked immunosorbent assay (ELISA). Formoterol increased the IL-6 release but did not influence the IL-8 release. Budesonide attenuated the IL 6 and IL-8 release, an inhibiting effect that was sustained, but not reinforced, by formoterol. PMID- 18415828 TI - Can equal access to care eliminate racial disparities in pediatric asthma outcomes? AB - A survey was given to the parents of 80 children with asthma between the ages of 3 and 18 years at the Pediatric Pulmonology Clinics of three military treatment facilities to evaluate asthma management and outcomes for different racial groups. Results demonstrated that management practices for the three groups were similar and that there were no significant differences in emergency department visits, prescription of oral steroids, or in the number of hospitalizations across the three groups. These findings suggest that equal access to care may allow children of different racial backgrounds to receive similar asthma care and achieve similar outcomes. PMID- 18415827 TI - Latino caregivers' beliefs about asthma: causes, symptoms, and practices. AB - BACKGROUND AND OBJECTIVE: This study examined belief systems of Latino caregivers who have children with asthma from Puerto Rican and Dominican backgrounds who resided on the Island of PR and the Mainland. The goal of this study was to document similarities and differences in beliefs about the causes, symptoms, and treatments of asthma across two sites and two Latino ethnic sub-groups of children who remain the most at risk for asthma morbidity. METHODS: Participants included 100 primary caregivers of a child with asthma; 50 caregivers from Island PR and 50 caregivers from mainland RI were interviewed (at each site; 25 caregivers were from Puerto Rican backgrounds and 25 caregivers were from Dominican backgrounds). The interview included an assessment of demographic information and beliefs about the causes and symptoms of asthma and asthma practices. RESULTS: Results indicated more similarities in beliefs about the causes and symptoms of asthma across site and ethnic group. The majority of differences were among beliefs about asthma practices by site and ethnic group. For example, a higher proportion of caregivers from Island PR, particularly those of Dominican descent, endorsed that a range of home and botanical remedies are effective for treating asthma. CONCLUSIONS: Results from this study point to several interesting directions for future research including larger samples of Latino caregivers with children who have asthma. A discussion of the importance of understanding cultural beliefs about asthma and asthma practices is also reviewed. PMID- 18415829 TI - Comparison of mometasone furoate dry powder inhaler and fluticasone propionate dry powder inhaler in patients with moderate to severe persistent asthma requiring high-dose inhaled corticosteroid therapy: findings from a noninferiority trial. AB - BACKGROUND: Inhaled corticosteroids (ICSs) are one of the suggested first-line therapies for patients with persistent asthma of moderate severity. METHODS: The efficacy and safety of mometasone furoate (MF) 400 microg twice daily (BID) and fluticasone propionate (FP) 500 microg BID administered for 12 weeks via dry powder inhaler (DPI) were compared in a noninferiority trial, in adults with moderate-to-severe persistent asthma. The primary variable was the change from baseline in am peak expiratory flow rate (PEFR). PM PEFR, forced expiratory volume in 1 second (FEV(1)), asthma symptoms, rescue medication use, response to therapy, exacerbation rates, and adverse events were also assessed. RESULTS: The lower bound of 95% CIs for treatment differences in the primary variable ranged from 2.6% to 5.6% throughout the 12-week study and were within the prespecified noninferiority range. No significant between-group differences were observed in lung function, rescue medication use, response to therapy, exacerbation rates, or adverse events. At most of the weeks assessed, there were no between-group differences in asthma symptoms. Most adverse events were mild-to-moderate. CONCLUSION: MF-DPI 400 microg BID was therapeutically equivalent to FP-DPI 500 microg BID in patients with moderate-to-severe persistent asthma. PMID- 18415830 TI - Psychological and somatic symptoms in screening for depression in asthma patients. AB - Correctly identifying asthma patients who need treatment for depression is part of comprehensive care. The objective of this study was to compare the prevalence of depressive symptoms measured by the short-form Center for Epidemiologic Studies Depression Scale (CESD-SF), which measures somatic and psychological symptoms, with the original and short-form Geriatric Depression Scale (GDS and GDS-SF), which measure only psychological symptoms. In total, 257 asthma patients (mean age 42 years, 75% women) completed the GDS (score range 0-30, positive screen > or = 11) and the CESD-SF (score range 0-30, positive screen > or = 10). The performance of each scale was compared to clinical diagnoses of depressive disorders reported by physicians using a skill score analysis. Twenty percent of patients had GDS scores > or = 11 and 32% had CESD-SF scores > or = 10. The somatic symptom of restless sleep was the most common CESD-SF symptom and the symptom that contributed most to the total score. The GDS had a skill score of +.16 (+1 = maximum possible, 0 = best guess) and the CESD-SF had a skill score of -.02 compared to physician-reported depressive disorders. Similar results were found for the GDS-SF. Thus, more patients had a positive CESD-SF screen, which was attributable mostly to a somatic sleep symptom that overlaps with asthma symptoms, and the GDS was more consistent with physicians' reports of depressive disorders. PMID- 18415831 TI - Effect of feedback letters to physicians and pharmacists on the appropriate use of medication in the treatment of asthma. AB - BACKGROUND: Suboptimal medication treatment of asthma has been reported. More specifically, short-acting beta 2-agonists are overused, while inhaled corticosteroids are underused. This can be related in part to poor adherence by patients to the prescribed regimen and to professionals' failure to comply with practice guidelines. Feedback seems to have an effect on professional practices related to medication use. OBJECTIVE: To assess the impact of feedback letters to physicians and pharmacists on their patients' appropriate use of asthma medication. METHODS: Two randomized trials were set up in the province of Quebec, Canada: one with physicians and another with pharmacists. A sample of voluntary physicians and pharmacists was randomly assigned to either the experimental group or to the control group. Those in the experimental groups received three consecutive feedback letters over a 9-month period summarizing the asthma medications acquired by their patients over the preceding year. The feedback focused on short-acting beta 2-agonists, long-acting beta 2-agonists and antileukotrienes and provided information on compliance with five appropriate-use criteria. Pharmacists received aggregate profiles and individual profiles with patients' names, while most physicians received aggregate profiles for all their eligible patients. Each mailing also included a pamphlet that summarized practice guidelines on asthma treatment. RESULTS: Seventy-one physicians and 60 pharmacists participated in the study. Physicians who received the feedback letters did not differ from those in the control group in terms of their proportion of prescriptions compliant with the criteria, either before the feedback or after it (p > 0.05). The before-after difference was also similar between groups. The same was true for pharmacists. However, although the before after difference for criteria 1 (frequency of use of short-acting beta 2 agonists) and 2 (frequency of use of long-acting beta 2-agonists) did not reach the usual statistical significance threshold of 0.05, the p value was under 0.10. CONCLUSIONS: As designed in this study, feedback provided to physicians did not improve the appropriate use of asthma medication. However, feedback to pharmacists is promising, especially when including patients' names so that pharmacists can intervene more specifically. PMID- 18415832 TI - Reduction of the total IgE level by omalizumab in children and adolescents. AB - BACKGROUND: Current data from clinical studies show that patients with severe allergic asthma experience a significant improvement from omalizumab. The early and late allergic reactions are inhibited by formation of complexes with free circulating immunoglobulin E (IgE), independent of which antigen activates the allergic cascade. The dosage of omalizumab depends on body weight and IgE level, yet no parameter has been established to guide dosage changes during therapy. The aim of this study was to investigate the value of the determination of total IgE by ADVIA Centaur assay to monitor the therapy progress. PATIENTS AND METHODS: Nine patients, 8 to 17 years of age, received therapy with omalizumab due to severe allergic bronchial asthma. In addition, the patients had pronounced rhinoconjunctivitis, food allergy, insect sting allergy, and/or neurodermitis. The total IgE in the serum (Sandwich-Immunoassay ADVIA Centaur) was measured in the patients once monthly before each omalizumab injection as a potential progress parameter. RESULTS: Six months after the beginning of therapy with omalizumab, a significant decrease of the total IgE concentration was found, in comparison to the baseline values (p < 0.01). In all patients, the tolerability of omalizumab was very good; there was a reduction in the frequency of the asthma exacerbations and rescue medications. The dosage of inhaled glucocorticoids could be lowered. All patients reported a clearly improved quality of life. CONCLUSIONS: The increase of the total IgE concentrations after administration of omalizumab described in the literature could not be confirmed. The value of total serum IgE as a progress parameter should be investigated in controlled studies with regard to sensitivity and specificity of the respective assays. The establishment of a test procedure for therapeutic monitoring appears urgently necessary, so that the appropriate dosage of omalizumab is applied in children and adolescents. Patients receiving omalizumab therapy should be closely monitored. PMID- 18415833 TI - Training improves physical fitness and decreases CRP also in asthmatic conscripts. AB - To study the respiratory and physical health of young men, 224 asthmatic and 668 non-asthmatic military conscripts were recruited from the intake groups of July 2004 and January 2005 in Kajaani, Finland. Factors affecting respiratory health were elicited by a questionnaire at the beginning of the service, and results of high sensitive C-reactive protein (hsCRP) determination, peak expiratory flow (PEF), and 12-minute running test were collected at the beginning and the end of the service. Respiratory infections were diagnosed by a study physician. Upon entering military service, asthmatics had frequent exercise- and cold-related asthma symptoms (69.6% and 76.3%), and 48% of them had no medication for asthma. At the beginning, 25.8% of asthmatics and 19.1% of non-asthmatics had a poor result of less than 2,200 m (p = 0.05) in the 12-minute running test, and after 180 to 362 days of service, the corresponding percentages were 11.7% and 9.7% (p = 0.434). The levels of hsCRP, a marker of low-grade systemic inflammation, decreased significantly among both asthmatics, 1.5 (p = 0.001), and non asthmatics, 1.6 mg/L (p < 0.001). Asthmatic men had 0.2 and non-asthmatics 0.1 respiratory infections per month (p < 0.001). In summary, asthmatic conscripts can enhance their physical fitness by training similarly to non-asthmatic ones. Their levels of hsCRP also decrease. PMID- 18415834 TI - The efficacy of montelukast and airway mast cell profiles in patients with cough variant asthma. AB - BACKGROUND: Cough variant asthma (CVA) is characterized by chronic cough without apparent wheezing; its pathophysiology is considered to be similar to that of classic asthma. OBJECTIVE: The clinical effects of montelukast, a cysteinyl leukotriene receptor antagonist, on cough variant asthma were assessed, and the activation profile of airway mast cells was examined. METHODS: Montelukast (10 mg/day) was given orally to 36 CVA patients (25 women and 11 men; median age, 37.5 years). Before treatment, the patients' bronchial mucosa underwent a biopsy with a fiberoptic bronchoscope. The biopsy specimens were double stained with anti-CD63 antibody and anti-human tryptase antibody. RESULTS: After 2 weeks of montelukast treatment, cough symptoms improved in 22 patients (the effective group) but did not improve in 14 patients (the ineffective group); in the ineffective group, the symptoms disappeared 2 weeks after they were switched to fluticasone propionate (400 microg/day) inhalation therapy. In the effective group, the time interval from the onset of symptoms to the initiation of treatment was significantly shorter than in the ineffective group. The bronchial mucosa biopsy specimens showed that the proportion of CD63-positive cells in tryptase-positive mast cells was significantly higher in the effective group than in the ineffective group; although the total numbers of mast cells were not different between the two groups. CONCLUSION: There is a subgroup of CVA patients in whom leukotrienes are closely involved in the pathogenesis of their chronic cough; activation of airway mast cells may be an essential feature in these patients. PMID- 18415836 TI - Fungal infections in hematopoietic stem cell transplant recipients. AB - Fungal infections have historically been, and remain important causes of transplant-related morbidity in recipients of hematopoietic stem cell transplant (HSCT). However, there have been notable changes in the epidemiology and outcomes of invasive fungal infections, induced by changes in the transplant procedures as well as supportive care. This review discusses invasive fungal infections in hematopoietic stem cell transplant recipients, with a focus on the host and the two most common infections, candidiasis and those caused by moulds. PMID- 18415837 TI - Experimental guinea pig model of dermatophytosis: a simple and useful tool for the evaluation of new diagnostics and antifungals. AB - The aim of this study was to establish a simple guinea pig model for the purpose of evaluating diagnostic principles and treatment modalities for dermatophytic infections. The following variables were evaluated; pre-treatment of the skin by shaving versus tape stripping, Microsporum canis or Trichophyton mentagrophytes test strains as etiologic agents, differences in inoculum concentrations, and inoculation with and without occlusion. The course of infection was evaluated clinically by redness and lesion scores and mycologically by microscopy, culture, and histopathology. The applicability of the model was evaluated with a recently developed diagnostic pan-dermatophyte PCR and antifungal treatment was tested with an oral solution of itraconazole, 10 mg/kg, once daily during days 3-14 of the test period. Pre-treatment of the skin with a manual razor was for practical reasons preferable to tape stripping. Inoculation under occlusion showed no advantage in the establishment of experimental infections. Infection severity showed some association with the inoculum concentration and subtype of T. mentagrophytes but not in studies involving M. canis. The establishment of dermatophytosis was confirmed by histopathology. Surprisingly, microscopy was found to be less sensitive than culture and the latter was as sensitive as pan dermatophyte PCR. Itraconazole significantly reduced lesion and redness score, with M. canis infections responding better to itraconazole treatment than those caused by T. mentagrophytes. In conclusion, we established a dermatophytosis animal model, which was proven useful for evaluating diagnostic methods and antifungal susceptibility testing. PMID- 18415838 TI - Morphological changes of the dermatophyte Trichophyton rubrum after photodynamic treatment: a scanning electron microscopy study. AB - Treatment strategies for superficial mycosis caused by the dermatophyte Trichophyton rubrum consist of the use of topical or oral antifungal preparations. We have recently discovered that T. rubrum is susceptible to photodynamic treatment (PDT), with 5,10,15-tris(4-methylpyridinium)-20-phenyl [21H,23H]-porphine trichloride (Sylsens B) as a photosensitizer. The susceptibility appeared to depend on the fungal growth stage, with PDT efficacy higher with microconidia when compared to mycelia. The aim of this study was to investigate, with the use of scanning electron microscopy, the morphological changes caused by a lethal PDT dose to T. rubrum when grown on isolated human stratum corneum. Corresponding dark treatment and light treatment without photosensitizer were used as controls. A sub-lethal PDT dose was also included in this investigation The morphologic changes were followed at various time points after the treatment of different fungal growth stages. Normal fungal growth was characterized by a fiber-like appearance of the surface of the hyphae and microconidia with the exception of the hyphal tips in full mycelia and the microconidia shortly after attachment to the stratum corneum. Here, densely packed globular structures were observed. The light dose (108 J/cm2) in the absence of Sylsens B, or the application of the photosensitizer in the absence of light, caused reversible fungal wall deformations and bulge formation. However, after a lethal PDT, a sequence of severe disruptions and deformations of both microconidia and the mycelium were observed leading to extrusion of cell material and emptied fungal elements. In case of a non-lethal PDT, fungal re-growth started on the remnants of the treated mycelium. PMID- 18415839 TI - Expression of genes in normal human monocytes in response to Aspergillus fumigatus. AB - The objective of these studies was to investigate genes of importance in the pathogenesis of Aspergillus infections. To do so, we employed microarray methodology to explore gene expression in human monocytes infected with Aspergillus conidia as compared with unstimulated monocytes and those stimulated with lipopolysaccharide (LPS) signaling through TOLL-like receptor 4 (TLR4). We found 997 (P 1 year), about 50 % of patients show a complete response. Adverse effects are generally mild but relapses occur more often compared to surgery. Conservative therapies including BTX are currently considered mostly as the first-line treatment. Among the surgical procedures, lateral sphincterotomy is the most effective treatment but shows higher incontinence and general morbidity rates than BTX. PMID- 18415899 TI - [External anal sphincter repair using the overlapping technique in patients with anal incontinence and concomitant pudendal nerve damage]. AB - BACKGROUND: No single surgical technique has so far emerged as the optimal approach to treat defects of the anal sphincter in patients with postpartum fecal incontinence. Our approach is to repair the external sphincter using the overlapping technique to optimize morphological and clinical outcome. The results were correlated with preoperatively determined pudendal nerve function. METHODS: Thirty-five patients were followed up for three years after repair of the external anal sphincter. The patients had grade 2 (n = 29) or grade 3 (n = 6) fecal incontinence. Nineteen (54 %) patients had a concomitant defect of the internal anal sphincter and 28 (80 %) had abnormal pelvic floor EMG findings. Before surgery, all patients underwent conservative treatment with biofeedback and electrostimulation. The muscle ends were overlapped with Vicryl 4-0 sutures. A standardized protocol was used for the perioperative management in all patients. RESULTS: Of the 35 patients who underwent overlapping repair of the external anal sphincter, 32 (91 %) had a satisfactory result at 3-year follow-up based on sonomorphological criteria. These 32 patients were continent for solid and liquid stools. Six of the 35 patients (17 %) continued to have flatus incontinence. Two (6 %) patients were improved and one patient (3 %) had unchanged incontinence. Pudendal nerve damage had no effect on the outcome of surgery. CONCLUSIONS: Our findings at 3-year follow-up show good results for the overlapping repair of the external anal sphincter in terms of morphology and clinical symptoms. This outcome depends on an adequate preoperative pelvic floor conditioning, optimal perioperative management, and use of a standardized operative technique. Surgical repair of the morphological defect is recommended even in patients with pudendal nerve damage. PMID- 18415900 TI - [Indications and results of sacral nerve stimulation in faecal incontinence]. AB - BACKGROUND: Sacral nerve stimulation (SNS) is an effective and less invasive treatment of faecal incontinence (FI). Patient selection has evolved from strict criteria to a more liberal approach, since temporary testing reliably predicts the efficacy of permanent stimulation in FI of various aetiologies. PATIENTS AND METHODS: From November 2005 until June 2007, we evaluated 20 consecutive patients (17 females, 3 males) with FI by percutaneous nerve evaluation (PNE), i. e., temporary stimulation. 13 patients proceeded to a permanent implantation of a pulse generator (3 bilateral generators). 11 patients with permanent stimulation were eligible for a minimum follow-up of 3 months. Median follow-up for this group was 10 (range 3-19) months. All patients provided bowel diaries, the disease-specific quality of life questionnaire of the American Society of Colon and Rectal Surgeons (ASCRS), and the Standard Short Form Health Survey Questionnaire (SF-36) at baseline, screening and at the follow-up. RESULTS: The aetiologies of the FI were pelvic floor insufficiency (n = 12), history of anterior resection (n = 3), history of surgery for disk prolaps (n = 2), sphincter disruption (n = 1), history of surgery for recto-vaginal fistula (n = 1), and idiopathic (n = 1). The mean number of incontinence episodes dropped from 9.9 to 1.3 during temporary testing (p = 0.02) and to 4.5 at last follow-up (p = 0.043). The quality of life assessment showed a significant improvement in the subscale embarrassment of the ASCRS (p = 0.043). There were 2 minor postoperative complications, and 1 medium-term failure of SNS treatment. CONCLUSION: SNS is a minimally invasive and effective treatment of FI. A pragmatic approach is justified due to the possibility of temporary testing and the low rate of complications. PMID- 18415901 TI - [The laparoscopic spleen-preserving approach to splenic cysts: advantages and limitations]. AB - INTRODUCTION: Due to the multiple functions of the spleen the preferred management of cystic non-parasitic lesions is nowadays laparoscopic partial splenectomy or decapsulation with preservation of the spleen. We have analysed our patients with non-parasitic cystic lesions and the current literature to weigh the benefits and complications of these methods in children and adults. PATIENTS AND METHODS: Laparoscopic partial splenectomy was performed in three children with dysontogenetic cysts. Laparoscopic marsupialisation was performed in one child and in three adults. RESULTS: Follow-up in the true cysts showed no recurrence in one patient and one residual cyst in the second. In the third patient, splenectomy was performed because there was not enough residual spleen to warrant preservation. In the group with post-traumatic cysts, no complications or recurrences were observed. DISCUSSION: The laparoscopic spleen-preserving approach for the treatment of non-parasitic cysts is feasible, but challenging. The main problem is a recurrence rate of more than 20 %. The reason for recurrence remains uncertain: it is probably due to different operative strategies, the morphology of the cysts or the presence of residual cysts. The recurrence rate in post-traumatic cysts is low on the basis of our own experience and a literature survey. CONCLUSION: The benefit of laparoscopic treatment in true non-parasitic splenic cysts has to be weighed against the rate of recurrence. A complete resection of the cysts should be attempted. In post traumatic cysts, laparoscopy offers a good minimally invasive treatment option. PMID- 18415902 TI - [Lay assessment of health care services using therapy for rectal cancer as an example]. AB - AIM: Multimodal treatment strategies make the assessment of health care services very difficult even for experts and almost impossible for lay persons. The aim of this project was to present complex data from scientific publications in a simplified way so that all essential information is preserved, but still assessable by lay persons and to compare their assessments with the recommendations of experts. METHODS: Using the surgical treatment of rectal cancer with or without preoperative radiation as an example, the aims of treatment as well as the "outcomes" (actually intended study endpoints) and "outputs" (surrogate parameters) were defined, identified and presented graphically for five key studies of neoadjuvant treatment. RESULTS: German lay persons (n = 59) favoured in the majority of the cases (93 %) surgical treatment without preoperative radiation. Lay persons assessed the results in a similar manner to other groups (physicians, health care workers, and health care politicians) and lay persons of other cultural backgrounds. Altogether, the participants (n = 152) favoured surgical treatment without preoperative therapy in 86 % of the cases (653 of 760). This lay assessment did not correlate with the assessments and recommendations of the scientific societies responsible for the guidelines. CONCLUSIONS: Complex scientific results can be prepared in such a way that their assessment by lay persons is feasible. Lay persons orientate their assessment according to the outcomes, while the recommendations of the guidelines are more directed by the outputs. These different viewpoints should be taken more into consideration for the development of guidelines than they are now. PMID- 18415903 TI - [Influenceable surgical and anesthesiological risk factors for the development of cardiac and pulmonary complications in laparoscopic surgery of the colon]. AB - INTRODUCTION: Due to the demographic shift in the age structure of the population, increasingly older, multimorbid patients are operated who have a substantially higher risk for the occurrence of intra- and postoperative complications. Apart from the identification of patient-referred, hardly influenceable risk factors, influenceable intraoperative surgical and anesthesiological risk factors have hardly ever been examined. The aim of this investigation was therefore to identify influenceable risk factors for the development of post-operative morbidity. METHODS: In a period of 44 months, we performed a laparoscopic colon resection in 157 men and 209 women with a mean age of 63 years. The ASA classification, POSSUM score, status of the anesthesiologist, change of the anesthesiologist, intraoperative monitoring, kind of anaesthesia, fluctuations of blood pressure and pulse during the operation, shock-index > 1, substitution of erythrocyte concentrates and FFPs as well as intraoperative surgical complications were documented prospectively. Postoperative general complications requiring therapy, in particular, cardiac and pulmonal problems as well as surgical complications, in particular, infections and hemorrhages, were documented. The data analysis was performed using the program package SPSS. RESULTS: Intraoperative monitoring was more frequently used in higher ASA stages, whereas for ASA stage IV no central venous line was used in 17 % and no arterial catheter was placed in 33 %. a similar tendency concerning the POSSUM score could not be determined. Patients cared for by junior surgeons exhibited cardiac complications in 6.7 % and 13.1 % had to be mechanically ventilated postoperatively versus 2 % of cardiac complications and 9 % mechanical ventilation among those managed by specialists. An increase in postoperative complications could also be found when a change in anesthesia took place. During treatment by an assistant in case of emergencies, in cases where intraoperative substitution of erythrocytes or an operation lasting more than two hours, more cardiac complications and a higher rate of mechanical respiration was observed than during treatment by a specialist. A mechanical respiration was significantly more necessary in higher ASA stages (p < 0.01), in an operation lasting more than 2 hours (p < 0.01), in cases with the occurrence of intraoperative bleeding complications (p < 0.01), procedures with a lower status of the anesthesiologist (p < 0.01) and in procedures with a change of the anesthesiologist (p < 0.05). CONCLUSION: Factors such as overweight, ASA classification or urgency cannot be changed. Surgical factors such as a standardisation of the operation technique with reduction of the operating time and careful staunching of bleeding can help to reduce postoperative complications. Anesthesiologists can also help by avoiding a change of the anesthesiologist as well as by preference of specialists in patients with higher ASA stages and in emergency cases. PMID- 18415905 TI - ["Intraoperative fluid therapy"]. PMID- 18415906 TI - [Intraoperative fluid management in pancreatic resections--the surgeon's view]. AB - Even though intraoperative fluid management during major intraabdominal surgery has frequently been addressed in the past, there is a lack of evidence-based recommendations. This report elucidates the topic from the surgeon's view. For the surgeon, the influence of larger fluid amounts on wound and anastomotic healing, bleeding complications and postoperative outcome (time of extubation, postoperative gastrointestinal function, hospital stay, etc.) is of interest. To clarify the question as to what a perioperative fluid regime should be composed of from a surgical point of view, data from the literature and our own studies were evaluated. The retrospective analysis of 98 pancreas resections that had been performed in our hospital revealed no significant differences concerning the occurrence of postoperative bleeding (8.2 %), wound infection (4.1 %), pancreatic fistula (9.4 %) and mortality (2.0 %) based on the administered intraoperative fluid amount. These results were comparable to those of other authors. The average intraoperatively infused fluid amount was 13.9 +/- 0.9 mL / kg / h. Catecholamines were administered in 74 % of all operations, while noradrenaline was used in 54 % of all cases. Although other factors might play a role in this setting, we can deduce from these data that application of a volume of 10-15 mL / kg / h has no negative influence on the outcome following pancreas resections and that the intraoperative fluid therapy should be targeted at these values. PMID- 18415907 TI - [Is a pediatric inguinal hernia appropriate to laparoscopic repair?--Some considerations and a comment on the article: Classen et al.: Laparoscopic inguinal herniorrhaphy in children--experiences in a tertiary referral medical center]. PMID- 18415908 TI - [Laparoscopic inguinal herniorrhaphy in children--experiences in a primary medical care center]. AB - AIM: Children undergoing laparoscopic herniorrhaphy may benefit from a minimally invasive access and the possible exploration of the contralateral groin. In this prospective feasibility study, the experiences and results after introduction of this method in a tertiary referral medical center are reported. METHODS: 50 laparoscopic herniorrhaphies were performed in 44 children (32 boys and 12 girls, range: 17 months-11 years) with open inner inguinal rings between January 2001 and March 2007. In 6 children (13.6 %) an additional contralateral open inner inguinal ring was detected. Laparoscopic access was obtained with a 5-mm laparoscope which was inserted transumbilically and two 2-mm needle holders were inserted through the inferolateral abdominal wall. The open inner inguinal rings were closed in 8 children using a resorbable Z-suture and in 36 children using a monofilamentous non-absorbable suture. Follow-up examinations included records of intra- and postoperative complications, recurrence rate and economic aspects. RESULTS: No intra- or perioperative complications occurred. Median operation time was 19 minutes. 43 / 44 operations were carried on an outpatient basis. Average material costs were about 20 Euro without any difference from the open access. During 36 months the follow-up rate was 91 %. Two children who had previously received an absorbable suture suffered from a recurring groin hernia within the first 6 months. CONCLUSION: Laparoscopic herniorrhaphy in children represents a safe and effective surgical method that can be recommended for tertiary referral medical centers with training facilities and experience in minimally invasive surgery. PMID- 18415909 TI - [Recurrence of Crohn's disease after surgery--causes and risks]. AB - BACKGROUND: In spite of the important role of conservative treatment, up to 90 % of all patients with Crohn's disease will undergo an operation during the course of their illness. Up to 50 % even need a second operation or further surgical procedures - with an increasing risk for perioperative complications. This study was designed to identify the risk factors for recurrence in patients with Crohn's disease and the influence of the primary operation. METHODS: Between 1986 and 2004, 412 patients with Crohn's disease required operative treatment. 218 underwent a primary procedure and 194 needed a reoperation. In particular, the indications for surgical treatment, the surgical procedures and the perioperative complications were registered and evaluated in the context of a possible recurrence of Crohn's disease. In this study, "recurrence" is defined as a reoperation because of Crohn's disease after a primary operation. RESULTS: The most common indications for a surgical treatment were stenosis (58.4 %) and fistulas (38.5 %). As the most frequent procedures, the ileocoecal resection and the partial resection of the small bowel were performed. Altogether, the complication rate was 11.5 %. The primary procedures (6.52 %) had less complications than the operations for a recurrence of Crohn's disease (17.70 %). The rate for the recurrence of Crohn's disease was 17.4 % after 5 years, 36.7 % after 10 years and 52.8 % after 15 years. Patients with fistulas as the indication for primary operation had the highest rate of recurrence (45 %). Patients with an isolated Crohn's lesion of the small intestine had a significantly higher risk for recurrence (59.5 %) than patients with lesions in the ileocoecal region or the colon. The anastomosis region (73 %) was the most common localisation for recurrence. CONCLUSION: On the basis of defined risk factors, patients with a high risk for recurrence can be identified. This is very important because of the higher risk for complications caused by reoperations compared to primary procedures. That is why interdisciplinary cooperation including postoperative care and optimal conservative treatment are absolutely essential. PMID- 18415910 TI - [The way to establish laparoscopic donor nephrectomy as the method of choice in live kidney donation--a single centre experience]. AB - INTRODUCTION: In contrast to the USA, laparoscopic donor nephrectomy is rarely practised in German transplant centres. Safety concerns and difficulties with the learning curve of this advanced laparoscopic procedure are the main obstacles to the establishment of this operation. PATIENTS AND METHODS: From 1998-2005, we performed laparoscopic kidney procurement in 50 live kidney donors on an intention to treat basis harvesting a total of 29 left and 21 right kidneys for transplantation. RESULTS: Negative adverse effects on the donor side were temporary nerve irritation (2 patients) and postoperative retroperitoneal hematoma. Reasons to convert to open nephrectomy were bleeding (2 patients) and adhesions (1 patient). On the recipient side, one kidney was lost due to renal vein thrombosis. Three patients required short-time dialysis after transplantation. All other kidney transplants worked without any problems. CONCLUSION: Laparoscopic donor nephrectomy is a safe procedure and has been established as the method of choice for live kidney donation in our hospital. PMID- 18415911 TI - [Prevention of type 2 diabetes--a missed opportunity]. PMID- 18415912 TI - [Validation of the German Diabetes Risk Score with metabolic risk factors for type 2 diabetes]. AB - BACKGROUND AND AIMS: The German Diabetes Risk Score (DRS) was developed at the German Institute of Human Nutrition from data of the European Prospective Investigation into Cancer and Nutrition (EPIC-Potsdam) in order to estimate the 5 year probability of developing type 2 diabetes, based on anthropometric measures and lifestyle as well as diet information. This study evaluated associations between the DRS and metabolic risk factors of type 2 diabetes for the purpose of further validating the risk score. METHODS: 2500 participants of the EPIC-Potsdam study were randomly selected, and glucose, HbA1c, triglycerides, HDL cholesterol, hs-C-reactive protein, and gamma-glutamyltransferase were determined. After exclusion of participants with known diabetes (n = 120), those for whom not all biomarkers could be determined (n = 92) and those with elevated plasma glucose concentrations (random glucose >or= 200 mg/dl or fasting glucose >or= 126 mg/dl, n = 9) or with low plasma glucose concentrations (< 50 mg/dl, n = 38), the data on 2223 participants (839 men and 1384 women) remained for analysis of which 640 (252 men and 388 women) were fasting at the time blood samples had been obtained. RESULTS: The DRS significantly correlated with all biomarkers (p < 0.001). Pearson correlation coefficients ranged from 0.25 for glucose and 0.45 for triglycerides. While glucose, HbA (1c), triglycerides, hs-CRP and gamma glutamyltransferase increased with increasing DRS points, HDL cholesterol was inversely associated. The mean concentrations of glucose, HbA (1c) and triglycerides for persons with < 300 DRS points were 83 mg/dl, 6.2 % und 62 mg/dl. In contrast, mean concentrations were 100 mg/dl for glucose, 6,9 % HbA (1c) und 171 mg/dl triglycerides among participants with >or= 700 DRS points. HDL cholesterol was considerably lower (37.6 mg/dl) among participants with >or= 700 DRS points than those with < 300 DRS points (55 mg/dl). There were also considerable differences in biomarker concentrations for hs-CRP and gamma glutamyltransferase in relation to the DRS. Associations were generally similar for men and women. CONCLUSIONS: Our data suggest that the DRS allows to detect persons with prediabetes with high sensitivity and specificity. Thus the DRS could be used as a routine medical checkup in programs for the prevention of type 2 diabetes. PMID- 18415913 TI - [Bilateral fractures of the humerus after hypoglycemic seizure]. AB - HISTORY: An 87 year-old woman with insulin dependent diabetes was referred to our hospital because she had sustained a generalized tonic-clonic seizure. Initial blood sugar concentration before hospital admission was 1.1 mmol/l. INVESTIGATIONS: Conventional radiography and computed tomography of both shoulders confirmed the diagnosis of bilateral proximal fractures of the humerus with posterior dislocation. The cause of the seizure was found to be iatrogenic because timing of application of the dual-acting insulin medication had been wrongly changed, and the patient had developed severe hypoglycemia. TREATMENT AND COURSE: After administration of glucose and a 7-day course of antibiotics for aspiration pneumonia, both fractures were treated surgically with hemiarthroplasty. After a four-week stay in hospital the patient was discharged to her nursing home, requiring much more care than before. CONCLUSION: The presentation of simultaneous bilateral fractures of the humerus is a rare but severe and almost pathognomonic complications of generalized tonic-clonic seizures, for example in the context of marked hypoglycemia. Appropriate monitoring and treatment of blood sugar levels in diabetics must be emphasized. PMID- 18415914 TI - [Purple urine bag syndrome]. PMID- 18415915 TI - [Nutrition therapy for gestational diabetes]. AB - Overweight and increased energy intake before conception are powerful risk factors in the development of gestational diabetes mellitus (GDM) and may also represent important determinants of the so-called fetal (mal-)programming, which may have long-term consequences for the health of the newborn. Thus, an adequate intake of energy and nutrients is of fundamental significance in the treatment of GDM, along with regular self-monitoring of blood glucose. This concept suffices in most cases to achieve the strict therapeutic goal of normoglycemia. However, because of a lack of data from interventional studies, there is uncertainty about the optimal macronutrient composition of the diet (carbohydrates, fat, protein) and meal distribution, as well as of the mode of calorie restriction in overweight and obese women with GDM. Varying the carbohydrate intake between 40 and 55 % of total energy intake appears to be acceptable and may be distributed across main meals and snacks. Thus, individualized nutritional treatment together with other specific lifestyle interventions are the principal components in the management of GDM. PMID- 18415916 TI - [Intensive insulin therapy]. PMID- 18415917 TI - [Early insulin administration in obese patients with type 2 diabetes makes sense- pro]. PMID- 18415918 TI - [Early insulin administration in obese patients with type 2 diabetes makes sense- contra]. PMID- 18415919 TI - [Young physicians' view on factors that increase the attractiveness of general practice]. AB - PURPOSE: The objectives of this study are to investigate young physicians' career plans at the end of their residency, and to assess what factors contribute in their view to improve the image of family medicine/general practice and make it an attractive career goal. METHODS: As part of a prospective study on career determinants in young physicians, begun in 2001, 534 residents were asked in 2007 about their aspired to professional career, their planned model of private practice, arguments against or for family medicine/general practice, respectively, and factors to improve the attractiveness of the latter. RESULTS: Of the study sample, 84 participants (42% men, 58% women) decided on family medicine (60% specialise in primary care, 40% in general internal medicine), 450 specialise in other medical specialties. Of the 534 study participants, 208 plan to work in a private practice, mostly in a group practice (88%). Of the future family physicians, 49% want to open their practice in an urban area, of the future specialists the respective number is 77%. Main reasons not to decide on family medicine are the uncertain development in health policy and the low income. The diversity of work, the broad spectrum of patients and the continuity in the doctor-patient relationship are named as factors in favour of family medicine. The possibility to run interdisciplinary group practices and better financial conditions are crucial factors to make family medicine an attractive career goal for young physicians. CONCLUSIONS: Family medicine is still assessed as an interesting field, however, the conditions of work as a family physician/general practitioner have a deterrent effect on young physicians. PMID- 18415920 TI - [Dementia patients and levels of care: do symptoms of the illness affect the grading?]. AB - OBJECTIVE: As the levels of care within long-term nursing care regulations are defined according to limitations in performing fundamental activities of daily living, the extent to which medically diagnosed cognitive, emotional and behavioral dementia symptoms are taken into account in the grading, should be investigated. METHOD: 390 patients with mild to moderate dementia from the Mid Franconia region, were included into the IDA ("Initiative Demenzversorgung in Allgemeinmedizin") study by specially trained general practitioners. The GPs had diagnosed dementia and noted the accompanying signs at baseline. In an interview with the caregiver, the Barthel Index was used to measure the level of help required with fundamental daily tasks. Predictors for grading were set down using logistic regression analysis. RESULTS: Where one accompanying sign is present, about half the patients had not been assigned a grade. Besides the Barthel Index, the presence of agitation and agnosia and the absence of depression are the only independent predictors for grading. All other symptoms, impairment of the executive function, loss of orientation, aphasia, anxiety, sleeplessness, aggressiveness and tendency to wander, have no significant predictive value. CONCLUSION: In the future development of nursing care insurance, the need for nursing care should be redefined using symptoms associated with dementia, particularly sleeplessness, aggressiveness and the tendency to wander as inclusion criteria. This is a prerequisite of improving the care available to dementia patients in the long term and also of expanding relief measures for family caregiver. PMID- 18415921 TI - [Health indicators and preventive behaviour of older employees in geriatric care]. AB - STUDY OBJECTIVE: As a consequence of the demographic transition, the proportion of geriatric nurses aged over 50 is increasing. The present study examines whether the prevalence of skin and back diseases and preventive behaviour differ between this group and younger employees. An additional objective was to record the proportion of older employees with a health- and occupation-related impairment. METHODS: 2 149 nurses working in home care settings and nursing homes were questioned in written form about their working ability, their preventive behaviour and their health situation. They were also subjected to dermatological investigation (with the Osnabruck Hand Eczema Severity Index - OHSI) and orthopaedic investigation (based on the multiple-step inventory of examinations). RESULTS: Older nurses more often rated their working ability as "(fairly) poor" than did younger nurses. This applied both to physical demands (11 vs. 4%) and to psychological demands (10 vs. 5%). They were also more affected by symptoms in the cervical spine (28 vs. 13%) and the lumbar spine (56 vs. 37%). Hand eczema was not more prevalent in older nurses. The state of health of 10% of the nurses aged over 50 indicated a health- and occupation-related impairment in regard to work ability and back complaints. Roughly equal proportions of the younger and older groups participated in measures for the primary prevention of back symptoms. The older nurses participated more often in advisory sessions on skin protection and more frequently applied cream to their hands. The younger nurses were in skin contact with water or fluids for shorter periods and more often exclusively used disinfectants after patient contact. CONCLUSION: Geriatric nurses with a health- and occupation-related impairment require intensive measures to support their health and to prevent health damage and to stop them from leaving work prematurely. If at all possible, these measures should not be restricted to a single social insurance agency. The individual preventive behaviour to avoid occupational back symptoms is not dependent on age. An age effect was observed for individual measures to protect the skin. Nevertheless, these had no effect on the risk of disease. The results indicate that more attention should be given to older employees during occupational training. PMID- 18415922 TI - [Social inequality as a predictor of occupational reintegration of chronic back pain patients following medical rehabilitation]. AB - BACKGROUND: The question of which factors influence return to work following medical rehabilitative measures in the case of chronic back pain is, in a social political context, of international significance. While psychosocial, socio economic and demographic factors as well as subjective perceptions of working conditions are individually discussed, the role of social inequality has thus far received little attention. OBJECTIVE: The present paper focuses on the occupational reintegration of patients with chronic dorsopathies following a medical rehabilitative intervention and aims to investigate the influence of predictor variables on successful reintegration. METHODS: The re-analysis conducted was based on a three-year prospective controlled intervention study with three measurement sessions. The study comprised a sample of 289 patients aged 20 to 64 years who had undergone first-time intervertebral disc operations in the lumbar spine region. Data were collected between March 2002 and February 2005 using a standardized written questionnaire. Winkler's (1998) composite index comprising indicators of occupational position, income and education was employed as a measure of social inequality. Age and sex were also included as independent variables in a binary logistic regression analysis. RESULTS: Of individuals undergoing rehabilitation, those from lower social classes returned to work later than those from higher social classes. Age and sex did not prove to be significant predictors of successful return to work. DISCUSSION: The probability of successful reintegration into working life increases according to the position within the vertical structure of social class. In light of this result, it can be assumed that not only differential allocation of patients to rehabilitation interventions but also the specific contents of these interventions should be modeled on the basis of economic criteria. PMID- 18415923 TI - [Psychosocial risk and protective factors for mental health in childhood and adolescence - results from The German Health Interview and Examination Survey for Children and Adolescents (KIGGS)]. AB - This report is based on the actual representative epidemiological data and describes the prevalence of mental disorders in children and adolescents in the context of selected risk and protective factors. The German Health Interview and Examination Survey for Children and Adolescents (KIGGS) showed that every 7th child aged 3-17 years had noticeable mental problems. At present we are discussing a multi-factorial developmental theory for these problems. It encompasses genetic and physiological factors together with personal and individual-developmental aspects as well as psychosocial and other environmental factors which have an influence on the development and progression of mental disturbances and illnesses. The realization of the potential of existing risk and protective factors is the base for the development of preventative measures to avoid mental dysfunction. Psychosocial risk factors in children and adolescents aged 3-17 years were identified as socio-economic status, single parent households and maternal unemployment. Children and adolescents with protective factors showed fewer signs of mental problems in comparison to their fellow age group with deficits of protective factors. These results show that personal resources are the most important predictors for mental health, followed by familial and social resources. PMID- 18415924 TI - [Predictors for compliance in orthodontic treatment]. AB - OBJECTIVE: Having established the importance of compliance as a significant factor of a successful orthodontic treatment, the aim of the present study was to evaluate intrapersonal and interpersonal factors which could help predict patient compliance. METHODS: The attributional style of 58 patients was assessed by a standardised questionnaire. An individual questionnaire was designed to determine attitudes concerning orthodontic treatment, the doctor-patient relationship, the wearing behaviour and control behaviour. The questionnaire was answered by the patient and by the orthodontist. The compliance was evaluated by the orthodontist on the basis of commonly used indicators for compliance: wearing time, oral hygiene and reliability of keeping the appointments. RESULTS: The results showed a significant correlation between the compliance and the attributional style of the patients in positive situations, but not between the compliance and the wearing behaviour estimated by the patient. The interpersonal comparison revealed a lack of knowledge on the part of the orthodontist about the patients' daily activities and their ability to correctly wear their appliances. CONCLUSION: The questionnaire answers show compliance to be a subjective construct of the orthodontist demonstrating mostly social-emotional matters. There is no consistency with compliance and the patients' statement concerning their wearing behaviour but with a positive attitude on the part of the patients demonstrating independent responsibility. PMID- 18415925 TI - [Outpatient management of stroke patients from the viewpoint of general practitioners in Hamburg--an exploratory study]. AB - OBJECTIVE: Stroke is the archetype of a chronic disease, with a considerable burden on ambulatory care. After discharge from hospital or inpatient rehabilitation, stroke patients are attended primarily by general practitioners (GPs). Little is known about the quality of outpatient care for stroke patients, so this study focused on the views of professionals involved in outpatient care. METHODS: A postal survey of 443 general practitioners from Hamburg, Germany was undertaken. Results were compared with a parallel survey carried out with physiotherapists and occupational therapists. RESULTS: 89 questionnaires were analysed. Based on a response rate of 28% the results cannot be seen as representative, but they do provide important information. As it seems, problems in ambulatory care do not start after discharge from hospital, but are partly caused by inpatient care. GPs see therapeutic benefits in ambulatory care for only half of their chronic stroke patients. They assess their own therapeutic efforts as effective in terms of medication, but they are less successful in changing their patients' behaviour with regard to lifestyle. Estimations of the prevalence of post-stroke depression vary substantially, with a mean of 47%. With regard to their daily practice, 60% of GPs do not find guidelines to be helpful. All in all, respondents describe the situation of chronic stroke patients and their families to be complex and difficult to manage. In addition to their professional performance, GPs carry out duties related to case management - as do the therapists. CONCLUSION: The care of stroke patients is not only the responsibility of a single group of professionals, but is a rather complex task involving different health-care professionals. The rehabilitation process and the interdisciplinary cooperation need to be further analysed to ensure a good cooperation between health-care professionals and to improve the quality of ambulatory care for stroke patients. PMID- 18415926 TI - [Amendment to the law governing the professional activities of physicians in general practice]. AB - The amendment to the law governing the activities of physicians in general free practice, which came into force on 1st January 2007, has brought a large number of changes. The age limit has been abolished, partial licences to practice and the formation of professional communities covering more than one district are permitted and new employment possibilities for physicians have been created. With these new regulations, many traditional principles of this law have been fundamentally changed. The purpose of the amendment is to make the health service more flexible and increase competition among providers of medical services. PMID- 18415927 TI - A new combined "interrupted-continuous" microvascular anastomotic technique. Experimental study and clinical application. AB - A new combined interrupted-continuous microvascular anastomotic technique is introduced, using four interrupted stay stitches and continuous suturing in between. Its efficacy is compared with the simple interrupted and continuous techniques. Microanastomoses were performed in the femoral artery and vein of rats. The patency, anastomosis time, and blood loss from the suture line of all anastomoses were recorded. Also, the sites of all anastomoses were harvested and histologically examined for lumen stenosis, media regeneration, and endothelial lining. Experimentation findings showed 100% patency rate in all groups. The combined and the continuous techniques were significantly faster and watertight compared with the simple interrupted. The only difference in pathology findings was the development of significant lumen stenosis in vein anastomoses performed with the continuous technique, compared with the other two techniques. The combined technique was clinically used in 16 free flap transfers and ensured 100% patency in all arterial and vein anastomoses, as well as survival of all flaps. In conclusion, the combined technique was found to be fast and easy to perform and ensured the same patency rate as the simple interrupted and continuous techniques. It also diminished the lumen stenosis effect of the continuous technique in vein anastomoses. Therefore, we suggest its use in both experimental studies and clinical cases. PMID- 18415928 TI - Radial forearm versus anterolateral thigh flap reconstruction after hemiglossectomy: functional assessment of swallowing and speech. AB - The authors retrospectively compared the results of postoperative speech and swallowing in patients who had undergone hemiglossectomy for carcinoma of the anterior tongue. Immediate reconstruction in 16 patients was with a free radial forearm flap and in another 16 with an anterolateral thigh flap. Clinical speech pathology evaluation included the Fletcher time-to-time maximum syllable repetition rate, multiple rhyme test, and overall quality and intelligibility of the patients' speech. Evaluation of swallowing included deglutition duration, bolus volume, and ingestion rate. The functional results with both flaps were adequate, and the two groups did not differ significantly between each other for either speech or swallowing. PMID- 18415929 TI - [Determinants of unplanned discharge from in-patient drug and alcohol detoxification: a retrospective analysis of 239 admissions]. AB - Unplanned and premature discharge from in-patient alcohol or drug detoxification is a common and severe problem in the treatment of substance abuse. So far, most of the relevant studies focused on drug detoxification, whereas only few studies also investigated alcohol detoxification. The aim of the present study was to comparatively identify and analyse determinants of unplanned discharge during in patient treatment in both diagnostic groups which simultaneously underwent detoxification under the same treatment setting. Subjects were 239 consecutive admissions (alcohol: n = 90; illegal drugs: n = 149) to a specialised qualified detoxification unit at the Psychiatric University Hospital of Heidelberg during the year 2000. Data on sociodemographical and psychosocial variables, medical history, psychopathological findings on admission and presence of psychiatric and/or somatic comorbidity as well as intensity level of withdrawal symptoms were collected retrospectively and analysed with respect to the prediction of planned/unplanned discharge. The high overall rates of unplanned discharge (alcohol: 43.3 % and drugs 62.4 %) confirm the previously reported figures. Treatment success of drug patients was rather affected by sociodemographical and psychosocial factors such as level of education, delinquency, unemployment and hepatitis C diagnosis. Relating to alcohol patients psychopathological findings on admission including orientation, affective state and cognition were most relevant for planned discharge. Furthermore, the results of this study underline the central role of motivation during in-patient treatment as well as the importance of a planned treatment continuation after discharge from the detoxification program. PMID- 18415930 TI - [Pharmacology of disulfiram - an update]. AB - Recently, we have been observing an increase in prescription of Disulfiram requiring medical knowledge concerning the overall effects, side effects and drug interactions of this substance. In the following report, we give an overview about the pharmacology and the side effects of Disulfiram. As an example we demonstrate a case, in which an acute neurological deficit was misjudged as a side effect of Disulfiram. Furthermore, the potential of Disulfiram as a promising treatment in addiction medicine will be discussed. PMID- 18415931 TI - Validity of a repeated-sprint test for football. AB - Three studies involving 108 football players were conducted to examine the reliability of a repeated-shuttle-sprint ability (RSSA) test and its ability to differentiate between players of various competitive levels and playing positions. Study 1: Short-term reliability was determined in 22 professional players completing the RSSA test (6 x 40-m sprints with 20 s of recovery between sprints) on two separate occasions. Study 2: Long-term reliability (seasonal changes) was examined in 31 professional players completing the RSSA test four times (during the preseason period, at the start, middle and end of the competitive season). Study 3: 108 players were divided and compared according to competitive level or playing position. Standard error of measurement values expressed as coefficient of variation for RSSA mean time and best time were 0.8 and 1.3 % (short-term reliability) and 0.9 and 1.2 % (long-term reliability), respectively. The smallest worthwhile changes were 0.5 % for both mean and best time. Professional players showed better RSSA performance than amateur players, and defenders displayed the lowest RSSA performance. In conclusion, the RSSA test showed adequate construct validity but only RSSA mean time showed sufficient reliability to detect large training-induced changes but not small important differences. PMID- 18415932 TI - Loss of Sox9 function results in defective chondrocyte differentiation of mouse embryonic stem cells in vitro. AB - The transcription factor Sox9 plays an important role during chondrogenesis. After early conditional inactivation of Sox9 in mesenchymal limb bud cells of mice, mesenchymal condensations as well as cartilage and bone are completely absent in the developing limbs. We analyzed chondrogenic differentiation of Sox9 /- mouse embryonic stem cells in vitro, using two clones with different targeted mutations. We found that the development of mature and hypertrophic chondrocytes is completely inhibited in the absence of Sox9 confirming that Sox9 is required for the formation of cartilage. In contrast, Sox9+/- mouse embryonic stem cells showed continuous but reduced differentiation into mature chondrocytes. Interestingly, the formation of early chondrogenic condensations expressing characteristic marker genes such as scleraxis, Sox5 and Sox6 was not inhibited in the absence of Sox9 in vitro. Thus, we propose that the earliest step of chondrogenesis could be regulated by a non cell-autonomous function of Sox9. PMID- 18415934 TI - Spatiotemporal clustering of cell death in the avian forebrain proliferative zone. AB - The extent to which programmed cell death is the fate of proliferative, rather than post-mitotic, cells remains controversial, but a preponderance of evidence suggests that at least some cells within the brain's proliferative zone die during mammalian brain development. One major unresolved question is the extent to which cell death in the proliferative zone is spatiotemporally patterned. In order to answer this question we used the terminal dUTP nick end labeling (TUNEL) method to stain apoptotic cells in the forebrain of chicken embryos at relatively early stages of brain development (Hamburger-Hamilton stages 19-32). Our principal finding is that most of the TUNEL-positive cells within the brain's proliferative zone are concentrated into distinct clusters, whose location varies with developmental stage. At stage 19, many TUNEL+ cells are found within the basal synencephalon, just below where the forebrain's first neurons are located. At stages 24-26, numerous TUNEL+ cells are located within the preoptic area and along the optic stalk. After stage 26, TUNEL labeling is prominent in two telencephalic areas: the thin dorsomedial telencephalon and the thickest portions of the telencephalon's lateral walls (i.e. the dorsal ventricular ridge). Collectively, the observed pattern of TUNEL staining suggests that cell death in the proliferative zone plays a substantial role in shaping the forebrain. In addition, cell death in the proliferative zone may be related to cell cycle exit. PMID- 18415933 TI - The distribution and behavior of extragonadal primordial germ cells in Bax mutant mice suggest a novel origin for sacrococcygeal germ cell tumors. AB - In the mouse, germ cells that do not reach the genital ridges rapidly die by a wave of apoptosis that requires the pro-apoptotic protein Bax. In Bax-null embryos, large numbers of ectopic (extragonadal) germ cells fail to die. We have studied the fates of these, in an effort to understand the etiology of human extragonadal germ cell tumors, which are thought to arise from ectopic germ cells. We find that ectopic germ cells in which apoptosis is blocked form a heterogeneous population, which partially differentiates along the gonocyte pathway to different extents in different regions of the embryo, and in the two genders. In particular, a previously undescribed population of ectopic germ cells was identified in the tail. These germ cells retained primitive markers for longer than ectopic germ cells in other regions, and represent a possible origin for sacrococcygeal type I extragonadal germ cell tumors found in neonates and infants. This hypothesis is supported, but not proved, by the finding of cells expressing the germ cell marker Oct4 associated with a coccygeal germ cell tumor in a human infant. PMID- 18415935 TI - Comparative study of mouse and human feeder cells for human embryonic stem cells. AB - Various types of feeder cells have been adopted for the culture of human embryonic stem cells (hESCs) to improve their attachment and provide them with stemness-supporting factors. However, feeder cells differ in their capacity to support the growth of undifferentiated hESCs. Here, we compared the expression and secretion of four well-established regulators of hESC pluripotency and/or differentiation among five lines of human foreskin fibroblasts and primary mouse embryonic fibroblasts throughout a standard hESC culture procedure. We found that human and mouse feeder cells secreted comparable levels of TGF beta 1. However, mouse feeder cells secreted larger quantities of activin A than human feeder cells. Conversely, FGF-2, which was produced by human feeder cells, could not be detected in culture media from mouse feeder cells. The quantity of BMP-4 was at about the level of detectability in media from all feeder cell types, although BMP-4 dimers were present in all feeder cells. Production of TGF beta 1, activin A, and FGF-2 varied considerably among the human-derived feeder cell lines. Low- and high-producing human feeder cells as well as mouse feeder cells were evaluated for their ability to support the undifferentiated growth of hESCs. We found that a significantly lower proportion of hESCs maintained on human feeder cell types expressed SSEA3, an undifferentiated cell marker. Moreover, SSEA3 expression and thus the pluripotent hESC compartment could be partially rescued by addition of activin A. Cumulatively, these results suggest that the ability of a feeder layer to promote the undifferentiated growth of hESCs is attributable to its characteristic growth factor production. PMID- 18415936 TI - Prolonged human/sheep cellular chimerism following transplantation of human hemopoietic stem cells into the ewe celomic cavity. AB - We evaluated the possibility of prolonged chimerism formation in fetus and lamb, following human cord blood-selected CD133+ hemopoietic stem cell (HSC) transplantation into the celomic cavity of ewes at a pre-immune fetal age (44-45 days of pregnancy). Nineteen ewes were injected with HSC and 5 controls with a saline solution. By PCR, HLA-DQ alpha 1 and 6 human microsatellites (CODIS) were used for HSC traceability. FISH analysis was performed with 8 human DNA probes from different chromosomes, to confirm chromosomal integrity, nuclear DNA localization and donor DNA identification. Immunological staining for revealing HLA-DQ alpha 1 expression demonstrated multilineage engraftment. Both HLA-DQ alpha 1 and microsatellites were detected in different tissues of 3 available aborted fetuses, to a lesser extent in 11 lambs tested at 2-months, but not 12 months after birth. Although only 1 fetus of siblings of each sheep was injected, all siblings revealed positive engraftments. Microsatellite analysis showed evidence of human allele segregation in different tissues of individual fetuses and lambs. FISH analysis confirmed chimerism and the presence of human chromosomes. Non-detection of some human gene sequences in different chromosomes and random finding of allele segregation for some human heterozygous microsatellites were found in different tissues of individual animals. Controls born from un-transplanted ewes never revealed any human DNA sequences nor HLADQ alpha 1 expression. PMID- 18415937 TI - Concentrations of TATA box-binding protein (TBP)-type genes affect chordamesodermal gene expression. AB - The TATA box-binding protein (TBP) is an essential component of transcription initiation complexes of all three eukaryotic RNA polymerases. Recent knockdown studies revealed that basic transcription factors are essential not only for gene transcription but also for regulating specific gene expression. However, the mechanism of and the effect by regulation of TBP expression are unknown during early embryogenesis. Here we show that the alteration of concentration of each TBP-type gene affected mutually one another's expression, suggesting that an optimal ratio of concentrations of TBP-type genes induce expression of specific genes. PMID- 18415938 TI - Expression of zinc transporter family genes in Dictyostelium. AB - Regulation of the zinc ion concentration is physiologically important to control the activities of a variety of cellular molecules. A BLAST search against a conserved domain of known zinc transporters identified twelve putative zinc transporter family genes in the Dictyostelium genome. Phylogenetic analysis revealed the presence of three zinc transporter subfamilies in Dictyostelium. One subfamily of proteins, consisting of the ZntA-D proteins, has weak homology to the STAT3-inducible LIV-1 protein. In addition, in situ hybridization revealed that the zntA-D genes are expressed in the pstAB cells, this expression being absent in the Dd-STATa null mutant. Thus, Dd-STATa may control stalk cell differentiation through some members of the zinc transporter family genes during Dictyostelium development. PMID- 18415939 TI - Calpain 2 expression pattern and sub-cellular localization during mouse embryogenesis. AB - Regulation of migration and proliferation by calpain has been shown in various cell types; however, no data are available concerning calpain 2 (capn2) localization in embryonic tissues. Here, we report the expression pattern of capn2 during mouse embryonic development. Expression of the capn2 gene is observed throughout embryonic development. From ES cells and the 8-cell stage to late neurulation stages, CAPN2 is expressed in the cytoplasm and nuclear compartments, with a clear co-localisation with chromatin. Whole-mount in situ hybridization analysis from E8.5 to 14.5 stages indicates high levels of capn2 expression in the nervous system, heart and mesodermal tissues. Up-regulation is maintained during later developmental stages in proliferating cells and in precursor cells involved in muscle (myoblasts) or bone formation (chondrocytes). At later developmental stages, elevated mRNA levels coincided with CAPN2 nuclear localization in these cell types, while differentiated cells maintained cytoplasmic expression. This detailed analysis reveals dynamic expression: nuclear localization was associated either with active cell mitosis in embryonic stem cells and early developmental stages or with precursor cells later during organogenesis. Thus, these data indicate that CAPN2 may represent a key factor in development from the first cell division. PMID- 18415940 TI - Expression and comparative genomics of two serum response factor genes in zebrafish. AB - Serum response factor (SRF) is a single copy, highly conserved transcription factor that governs the expression of hundreds of genes involved with actin cytoskeletal organization, cellular growth and signaling, neuronal circuitry and muscle differentiation. Zebrafish have emerged as a facile and inexpensive vertebrate model to delineate gene expression, regulation, and function, and yet the study of SRF in this animal has been virtually unexplored. Here, we report the existence of two srf genes in zebrafish, with partially overlapping patterns of expression in 3 and 7 day old developing animals. The mammalian ortholog (srf1) encodes for a 520 amino acid protein expressed in adult vascular and visceral smooth muscle cells, cardiac and skeletal muscle, as well as neuronal cells. The second zebrafish srf gene (srf2), encoding for a presumptive protein of only 314 amino acids, is transcribed at lower levels and appears to be less widely expressed across adult tissues. Both srf genes are induced by the SRF coactivator myocardin and attenuated with a short hairpin RNA to mammalian SRF. Promoter studies with srf1 reveal conserved CArG boxes that are the targets of SRF-myocardin in embryonic zebrafish cells. These results reveal that SRF was duplicated in the zebrafish genome and that its protein expression in all three muscle cell types is highly conserved across vertebrate animals suggesting an ancient code for transcriptional regulation of genes unique to muscle cell lineages. PMID- 18415941 TI - Expression of protocadherin 18 in the CNS and pharyngeal arches of zebrafish embryos. AB - Here, we report the results of molecular cloning and expression analyses of a non clustered protocadherin (pcdh), pcdh18 in zebrafish embryos. The predicted zebrafish pcdh18 protein shows 6566% identity and 7879% homology with its mammalian and Xenopus counterparts. It has a Disabled-1 binding motif in its cytoplasmic domain, which is characteristic of pcdh18. Zebrafish embryos expressed pcdh18 by the early gastrula stage, 6 h post-fertilization (hpf), in their animal cap but not in the germ ring or the shield. pcdh18 was expressed in the neural tube and the central nervous system (CNS) from 12 hpf. Some populations of cells in the lateral neural tube and spinal cord of 1218 hpf embryos expressed pcdh18, but expression in these cells disappeared by 24 hpf. The hindbrain of embryos at 2456 hpf expressed pcdh18 in cells closely adjacent to the rostral and caudal rhombomeric boundaries in a thread-like pattern running in the dorsoventral direction. The pcdh18-positive cells were localized in the ventral part of the hindbrain at 24 hpf and in the dorsal part from 36 hpf. pcdh18 was also expressed in the telencephalon, diencephalon, tectum, upper rhombic lip, retina and otic vesicle. Expression in the CNS decreased markedly before hatching. Pharyngeal arch primordia, arches, jaws and gills expressed pcdh18, and the molecule was also expressed in some endodermal cells in late embryos. PMID- 18415942 TI - Efficacy of pyriprole topical solution against the cat flea, Ctenocephalides felis, on dogs. AB - Three studies evaluating various aspects of the performance of pyriprole against the cat flea, Ctenocephalides felis, on dogs demonstrated that 12.5% pyriprole applied as a spot-on provides rapid, long-lasting efficacy against adult cat fleas, even under severe flea challenge. Speed of kill data indicate treatment with this product can interrupt an already established adult flea infestation, whereas monthly treatment can prevent reinfestation. Pyriprole disrupts the flea life cycle by killing adult fleas before they lay eggs for at least 30 days after treatment. The residual effect of pyriprole on debris from treated dogs (dander, hair, scales, and flea feces) resulted in a decreased ability of cat flea larvae to complete development to the adult stage for 2 weeks after application. Based on the results of these studies, 12.5% pyriprole represents a valuable new tool in the control of the cat flea, C. felis, on dogs. PMID- 18415943 TI - Efficacy of fipronil (9.8% w/w) + (S)-methoprene (8.8% w/w) and imidacloprid (8.8% w/w) + permethrin (44% w/w) against Dermacentor variabilis (American dog tick) on dogs. AB - This study evaluated overall efficacy, percentage of dogs free of live ticks, retention of ticks, and efficacy against retained ticks of fipronil (9.8% w/w) + (S)-methoprene (8.8% w/w) and imidacloprid (8.8% w/w) + permethrin (44% w/w) spot on topical solutions against Dermacentor variabilis-infested dogs. Tick control was significantly greater (P < .05) on dogs treated with fipronil + (S) methoprene than on control dogs for the entire 30-day study period. Conversely, the combination product of imidacloprid + permethrin demonstrated activity significantly (P < .05) different from that seen on control dogs only up to day 23. Significantly (P < .05) more dogs treated with fipronil + (S)-methoprene were free of live ticks compared with controls on days 3, 9, and 16. There was never a significant difference (P > .05) between control and imidacloprid + permethrin treated dogs in numbers of dogs free of live ticks. After the initial 10-minute tick exposure, lower numbers of ticks were retained on the imidacloprid + permethrin-treated dogs than on the fipronil + (S)-methoprene-treated dogs. However, when evaluating the effectiveness of the acaricides on "retained ticks," it appears that while some of the ticks were rapidly knocked down on the imidacloprid + permethrin-treated dogs, efficacy against ticks still retained at 10 minutes was never greater than 50%. PMID- 18415944 TI - Clinical field study of the safety and efficacy of spinosad chewable tablets for controlling fleas on dogs. AB - Preliminary studies showed spinosad to be rapidly effective and safe in controlling fleas on dogs. To validate these studies, a clinical trial was undertaken using 470 flea-infested client-owned dogs allocated to receive three monthly treatments with either beef-flavored chewable spinosad tablets (30-60 mg/kg) or selamectin applied according to label instructions. Flea counts 15 days after enrollment were reduced by 98.6% and 90.9% for spinosad- and selamectin treated dogs, respectively; at 90 days, flea count reductions were 99.9% and 98.9%, respectively. Compared with baseline, all flea reductions were significant (P < .001) for both products and spinosad was significantly (P <= .0172) more effective than selamectin at each postenrollment flea count. PMID- 18415945 TI - Serum pancreatic lipase immunoreactivity concentrations in dogs treated with potassium bromide and/or phenobarbital. AB - Potassium bromide, phenobarbital, or a combination of both is commonly used in the treatment of canine epilepsy. Several cases of clinical pancreatitis have been reported in dogs after treatment with potassium bromide, but the risk of elevated serum canine pancreatic lipase immunoreactivity concentrations in dogs treated with potassium bromide and/or phenobarbital has not previously been evaluated in a large group of dogs. This study suggests an increased risk for elevated serum canine pancreatic lipase immunoreactivity concentrations and possibly pancreatitis in dogs treated with potassium bromide or phenobarbital alone or in combination. PMID- 18415946 TI - Comparison of the time required to administer three different fluke and worm combination products to commercial beef cattle at housing. AB - Larger livestock units, a decline in the farm labor force, animal welfare concerns, and a trend toward more selective use of drugs have increased the focus on animal handling, time management, convenience, and compliance in administering veterinary therapeutics. This study was undertaken to quantify and compare the time needed to treat commercial beef cattle with three fluke and worm combination products with different administration profiles. Young beef cattle (n = 270) weighing approximately 400 kg were allocated to batches of five, which were randomly assigned to receive ivermectin + clorsulon injection, ivermectin + closantel injection, or levamisole + triclabendazole oral drench. The mean time needed to administer ivermectin + clorsulon (single injection) to five cattle was 31 seconds, which was significantly less than the 100 seconds needed for ivermectin + closantel (two injections) and the 126 seconds needed for levamisole + triclabendazole (P < .001). Such quantitative data can allow for better planning and selection of parasiticide treatment approaches at the farm level. PMID- 18415947 TI - Evaluation of on-arrival versus prompted metaphylaxis regimes using ceftiofur crystalline free acid for feedlot heifers at risk of developing bovine respiratory disease. AB - This study was designed to compare morbidity and mortality due to bovine respiratory disease (BRD) in 1,400 ranch-fresh, northern US calves that received either no metaphylaxis (MTX) (Group 1), MTX prompted by pen morbidity (Group 2), MTX prompted by a decline in feed intake (Group 3), or on-arrival MTX (Group 4). Ceftiofur crystalline free acid (6.6 mg/kg) was the MTX antimicrobial used. Decreased feed intake did not meet the study threshold, so Groups 1 and 3 were combined into a single control group. The percentage of calves not treated for BRD was 14% to 15% higher in Groups 4 and 2 (72.7% and 73.4%, respectively) compared with the controls (58.2%; P <= .02). Mortality was numerically higher in Group 2 (3.4%) compared with Groups 4 and 1 (0.9% and 1.6%, respectively). There were no differences in average daily gain or feed consumption. In northern, ranch fresh calves, MTX as a health management practice reduces the incidence of BRD and has better utility when performed on arrival versus relying on morbidity as a signal for antimicrobial administration. PMID- 18415948 TI - Psychotherapy termination: clinical and ethical responsibilities. AB - The successful termination of the psychotherapy relationship is each psychotherapist's goal. Yet, a number of circumstances may arise that interfere with the successful completion and termination of treatment. This article addresses both termination and abandonment and illustrates the applicable guidelines and standards from the American Psychological Association's (2002) Ethics Code. We conclude with 12 recommendations for proactively addressing termination and abandonment consistent with professional standards and each patient's best interests. PMID- 18415949 TI - Do syndromes matter (DSM)? PMID- 18415950 TI - Classification of anxiety and depressive disorders: problems and solutions. AB - The American Psychiatric Association and the World Health Organization have begun to revise their classifications of mental disorders. Four issues related to these revisions are discussed in this study: the structure of the classifications, the relationship between categories and dimensions, the sensitivity of categorical thresholds to definitions, and maximizing the utility and validity of the diagnostic process. There is now sufficient evidence to consider replacing the present groupings of disorders with an empirically based structure that reflects the actual similarities among disorders. For example, perhaps the present depression and anxiety disorders would be best grouped as internalizing disorders. Most mental disorders exist on a severity dimension. The reliability and validity of the classification might be improved if we accepted the dimensional nature of disorders while retaining the use of categorical diagnoses to enhance clinical utility. Definitions of the thresholds that define categories are very susceptible to detail. In International Classification of Diseases 11(ICD-11) and Diagnostic and Statistical Manual of Mental Disorders-V (DSM-V), disorders about which there is agreement should be identically defined, and disorders in which there is disagreement should be defined differently, so that research can identify which definition is more valid. The present diagnostic criteria are too complex to have acceptable clinical utility. We propose a reduced criterion set that can be remembered by clinicians and an enhanced criterion set for use with decision support tools. PMID- 18415951 TI - Hierarchical structures of affect and psychopathology and their implications for the classification of emotional disorders. AB - The Diagnostic and Statistical Manual of Mental Disorders-IV groups disorders into diagnostic classes on the basis of the subjective criterion of "shared phenomenological features." The current mood and anxiety disorders reflect the logic of older models emphasizing the existence of discrete emotions and, consequently, are based on a fundamental distinction between depressed mood (central to the mood disorders) and anxious mood (a core feature of the anxiety disorders). This distinction, however, ignores subsequent work that has established the existence of a general negative affect dimension that (a) produces strong correlations between anxious and depressed mood and (b) is largely responsible for the substantial comorbidity between the mood and anxiety disorders. More generally, there are now sufficient data to eliminate the current rational system and replace it with an empirically based taxonomy that reflects the actual-not the assumed-similarities among disorders. The existing structural evidence establishes that the mood and anxiety disorders should be collapsed together into an overarching superclass of emotional disorders, which can be decomposed into three subclasses: the distress disorders (major depression, dysthymic disorder, generalized anxiety disorder, posttraumatic stress disorder), the fear disorders (panic disorder, agoraphobia, social phobia, specific phobia), and the bipolar disorders (bipolar I, bipolar II, cyclothymia). An empirically based system of this type will facilitate differential diagnosis and encourage the ultimate development of an etiologically based taxonomy. PMID- 18415952 TI - Late percutaneous coronary intervention for the totally occluded infarct-related artery: a meta-analysis of the effects on cardiac function and remodeling. AB - BACKGROUND: Late percutaneous coronary intervention (PCI) of a totally occluded infarct-related artery (IRA) in stable patients is currently not recommended based on the lack of clear clinical benefits in randomized controlled trials. We sought to perform a systematic review and meta-analysis of randomized controlled trials comparing PCI with optimal medical therapy in patients with IRA occlusion more than 12 hr after onset of acute myocardial infarction (AMI), focusing on left ventricular function and remodeling. METHODS AND RESULTS: PubMed, CENTRAL, and mRCT were searched for eligible studies. Studies were included in the analysis if they were randomized controlled trials comparing conservative medical management with PCI performed at least 12 hr after the onset of symptoms of AMI, and data on left ventricular ejection fraction (LVEF) at baseline and follow-up were available. Studies were excluded if randomization occurred less than 12 hr after symptom onset, or if patients were hemodynamically unstable. Change in LVEF was the primary outcome of interest, with changes in left ventricular end diastolic volume index (LVEDVI) and end-systolic volume index (LVESVI) analyzed as secondary endpoints. We retrieved five studies in which baseline and follow up LVEF data were available enrolling a total of 648 patients: 342 patients randomized to PCI and 306 to medical treatment. There was a statistically significant difference in LVEF changes over time favoring PCI (+3.1%, 95% CI +1.0 to +5.2, P = 0.0004). In addition, there were statistically significant differences changes in both LVEDVI (-5.1 ml in favor of PCI, 95% CI of -9.4 to 0.8, P = 0.020) and LVESVI (-5.3 ml in favor in PCI, 95% CI of -8.3 to -2.4, P = 0.0005). CONCLUSIONS: This meta-analysis suggests that late revascularization of an occluded IRA may improve left ventricular systolic function and remodeling, supporting the "open artery hypothesis." The reason why these changes have not resulted in clinical benefits in large clinical trials is subject to debate. PMID- 18415953 TI - Late percutaneous recanalization of totally occluded infarct-related arteries- more than about time. PMID- 18415954 TI - Percutaneous coronary intervention for chronic total occlusion: novel 3 dimensional imaging and quantitative analysis. AB - OBJECTIVE: The purpose of this study was to evaluate chronic total occlusion (CTO) lesions in three dimensions (3D) and to examine the potential yield and added diagnostic value of these reconstructions in planning percutaneous interventional procedures. METHODS AND RESULTS: The novel CardiOp-B system for 3D reconstruction of the coronary vessels was used in 302 angiographic images from 58 consecutive patients (86% men; mean age 62 +/- 11 years) undergoing interventional treatment for CTO (61 CTOs). The success rate of 3D reconstruction was 83%. When successful, these reconstructions led to a significant improvement in lesion analysis, especially at the stump area and/or missing segment. Importantly, in 92% of the successful 3D reconstructions, the artery path in the lesion area could be delineated. In 95% of cases, in which post-stenting 3D reconstruction performed, the vessel path was similar to the lesion path suggested before stenting. The mean stenosis area was significantly smaller in the 3D reconstructions vs. 2D images (94 +/- 5.1% vs. 99 +/- 0.3%, P < 0.001), and the mean lesion length was significantly shorter (15.3 +/- 7.4 mm vs. 20.9 +/ 8 mm, P < 0.001). CONCLUSIONS: In most cases, 3D reconstruction of CTOs can clearly image the stump area, delineate the lesion path, and provide enough information for the clinician to precisely calculate the severity of stenosis and lesion length. 3D reconstructions may serve as a useful tool for planning interventional procedures for CTOs and improving their success rate. PMID- 18415955 TI - FDA notifications. FDA revises Baraclude labeling for HIV patients. PMID- 18415956 TI - FDA notifications. Tentative approval granted of generic nevirapine. PMID- 18415957 TI - Case: genotype reveals unusual discrepancy. PMID- 18415958 TI - Clinicians can help speed up Social Security adjudication process. PMID- 18415959 TI - FDA notifications. Tentative approval for generic stavudine, oral solution. PMID- 18415960 TI - FDA notifications. FDA approves NDA for Lexiva. PMID- 18415961 TI - Conscientious commitment. PMID- 18415962 TI - [Retropharyngeal tendinitis--a rare differential diagnosis of retropharyngeal abscess]. AB - Because of their similarity, clinical symptoms of a calcific retropharyngeal tendinitis can be misdiagnosed for a retropharyngeal abscess. We present two patients who were admitted for retropharyngeal abscess, however, careful examination revealed amorphous calcification anterior to C1-C2 in addition to prevertebral soft tissue swelling. Knowledge of the characteristic clinical symptoms and the pathognomonic imaging features is crucial to correctly diagnose the disease and to prevent unnecessary tests and treatment. PMID- 18415963 TI - [Interventions on the nose--management of acute nasal injuries; interventions of the nasal septum I]. PMID- 18415965 TI - [Information about anosmia, epidemiology, diagnosis, therapy]. PMID- 18415964 TI - [What is the procedure in expert assessment of hearing damage?]. PMID- 18415966 TI - Generic quality-of-life assessment in rheumatoid arthritis. AB - BACKGROUND: Generic health status measures are commonly used in the evaluation of rheumatoid arthritis (RA) patients. The reliability, validity, and sensitivity of the instruments in the assessment of quality of life (QOL) in RA, and how they correlate to other clinical measurements, have longbeen questioned. OBJECTIVE: Analyze the performance of a commonly used generic health status measure, the Medical Outcomes Study 36-Item Short Form (SF-36), against the Outcome Measures in Rheumatology(OMERACT) criteria. METHODS: Data were analyzed from 7 double blind, randomized controlled trials that examined the effectiveness of 1 or more interventions in RA. The primary outcome measures evaluated were the Mental and Physical Component Scores of the SF-36. Comparators were 1 or more of the following: the Health Assessment Questionnaire scores, tender joint count (TJC), the Disease Activity Score, and the American College of Rheumatology Responder Index (ACR20,ACR50, ACR70). The ability to detect a treatment effect in the study outcomes was evaluated using 3 measures: treatment difference,standardized response mean, and relative efficiency in relation to the TJC. RESULTS: As a generic QOL measure, the SF-36 is better suited to capture the holistic health of the patient, as reflected in the World Health Organization definition of health as being not only the avoidance of disease but the physical,emotional, and social well-being of the patient. Furthermore, use of the SF-36 permits comparisons of physical and mental aspects of QOL in the RA patient population, as well as comparisons of QOL parameters between patients with RA, other patient groups, and the general population. CONCLUSION: The SF-36 deserves serious consideration for inclusion in the core set of outcomes in RA trials. PMID- 18415967 TI - Improved health-related quality of life with effective disease-modifying antirheumatic drugs: evidence from randomized controlled trials. AB - Rheumatoid arthritis (RA) is a chronic autoimmune disease characterized by inflammation of the articular synovium, resulting in bony erosions, deformity, and, ultimately, joint destruction. With associated comorbid conditions,especially cardiovascular, it can result in significant morbidity as well as early mortality. Patients with RA report impairments in health-related quality of life (HRQOL) in comparison with age- and sex-matched populations without arthritis. These decreases in HRQOL are attributed to the pain, impairment in physical function, and fatigue associated with this disease. The introduction of new disease-modifying antirheumatic drugs has revolutionized the treatment of RA, particularly the biologic agents: etanercept, infliximab,adalimumab, abatacept, and rituximab.Importantly, administration of these agents has resulted in statistically significant and clinically meaningful improvements in physical function and HRQOL. Many clinical studies confirm that with these therapies, RA patients report improvements in HRQOL, reflected by improved physical function, less fatigue, and better emotional and mental function. Maintenance of physical function is no longer the only treatment goal for RA but also to improve, restore, and preserve HRQOL. Results from pivotal clinical trials are analyzed in this article and the relevance of the data derived from the clinical studies to day-to-day clinical practice are also discussed. PMID- 18415968 TI - [This week in the Wien Klischen Wochenschrift]. PMID- 18415969 TI - [System-forming factor of extrabrain changes in development of edema-swelling of the brain]. PMID- 18415970 TI - [Effect of lymphostimulation on lymphatic drainage of the rat lower limb in central lymphostasis]. AB - To evaluate functional status of rat hindleg lymphatic drainage in dynamic experiments, Wistar rats were divided into three groups: control, experimental and a group exposed to induced lymphostimulation. Central lymphostasis was provoked in experimental and lymphostimulation group. The latter group was also exposed to lymphotropic lymphostimulation. Functional status of the lymphatic drainage was assessed by the time of appearance of the marker (methylene blue) injected into the rat paw. The results indicate that lymphatic drainage and a carrier function of the lymph node were suppressed in the experimental group during the observation time. Lymphostimulation in the third group corrected the above disorders by experiment day 30. Thus, lymphostimulation can be an alternative method of management of the central lymphostasis syndrome in clinical practice. PMID- 18415971 TI - [Possibility of inverse development of morphological changes in the rat myocardium provoked by large doses of dopamine]. AB - It is shown that Wistar male rats keeping without medication and stressful effects for a month after administration of big doses of dopamine for two weeks does not lead to complete inverse development of heart remodeling. A decrease of the heart's mass occurs mainly because of a decrease of the right ventricle mass. The greatest changes of the extracellular matrix take place in the right ventricle: collagen level grows more noticeably, edema of the extracellular space relieves, correlations between collagen and blood vessels, cardiac hystiocytes and blood vessels increase aggravating right ventricular remodeling. PMID- 18415972 TI - [Mechanisms underlying changes in functional properties of red cells in acute action of carbon monoxide]. AB - It has been experimentally established that a single impact of carbon mono-oxide in concentration 4000 mg/m3 during 75 minutes (CL50) in rats is accompanied with disturbance of functional activities of blood red cells in the form of a decrease of deformability and an increase of aggregative capacity of erythrocytes which are most evident to the end of the first day after carbon mono-oxide administration. These changes are combined with structural modification and changes in metabolism of erythrocytic membrane. The signs of structural modification of erythrocytic membrane in rats exposed to acute effect of carbon mono-oxide in a mean lethal concentration stay long remaining during elimination of carboxyhemoglobinemia (up to 14-21 days). PMID- 18415973 TI - [Teaching pathophysiology in conditions of reforms of higher medical education]]. PMID- 18415974 TI - N-heterocyclic carbene ligands bearing hydrophilic and/or hydrophobic chains: Rh(I) and Pd(II) complexes and their catalytic activity. AB - A series of novel imidazolium salts bearing hydrophilic tetraethylene glycol (TEG) and/or hydrophobic long-chain alkyl (n-C12) functionalities, which are precursors for desired N-heterocyclic carbene (NHC) ligands, were synthesized and characterized. Rh(I)-NHC complexes were prepared in good yields by the silver carbene transfer method with NHC-Ag species derived from the imidazolium salts. The molecular structure of the Rh(I)-NHC complex having n-C12 chains has been determined by a single-crystal X-ray diffraction study and the complex is found to possess extended alkyl chains with anti conformations in the solid state. Hydrosilylation with the Rh(I) complexes and Suzuki-Miyaura coupling reactions with the Pd(II) complexes with these NHC ligands were carried out. In the latter case, the TEG moiety enhances catalytic activity considerably. PMID- 18415975 TI - Quality assessment of the Affymetrix U133A&B probesets by target sequence mapping and expression data analysis. AB - Careful analysis of microarray probe design should be an obligatory component of MicroArray Quality Control (MACQ) project [Patterson et al., 2006; Shi et al., 2006] initiated by the FDA (USA) in order to provide quality control tools to researchers of gene expression profiles and to translate the microarray technology from bench to bedside. The identification and filtering of unreliable probesets are important preprocessing steps before analysis of microarray data. These steps may result in an essential improvement in the selection of differentially expressed genes, gene clustering and construction of co-regulatory expression networks. We revised genome localization of the Affymetrix U133A&B GeneChip initial (target) probe sequences, and evaluated the impact of erroneous and poorly annotated target sequences on the quality of gene expression data. We found about 25% of Affymetrix target sequences overlapping with interspersed repeats that could cause cross-hybridization effects. In total, discrepancies in target sequence annotation account for up to approximately 30% of 44692 Affymetrix probesets. We introduce a novel quality control algorithm based on target sequence mapping onto genome and GeneChip expression data analysis. To validate the quality of probesets we used expression data from large, clinically and genetically distinct groups of breast cancers (249 samples). For the first time, we quantitatively evaluated the effect of repeats and other sources of inadequate probe design on the specificity, reliability and discrimination ability of Affymetrix probesets. We propose that only functionally reliable Affymetrix probesets that passed our quality control algorithm (approximately 86%) for gene expression analysis should be utilized. The target sequence annotation and filtering is available upon request. PMID- 18415976 TI - Simulation of coevolution in community by using the "Evolutionary Constructor" program. AB - An original modeling tool called Evolutionary Constructor has been proposed and described. Evolutionary Constructor combines the advantages of both generalized and portrait modeling and, additionally, provides an option to modify a current model's structure. The evolution of communities comprising atrophic ring-like network with the horizontal transfer of metabolism genes occurring among the communities has been modeled and presented. It has been demonstrated that a prolonged increase in the fitness of any single population that forms part of that trophic ring-like network of antagonistic communities will eventually lead that system to becoming absolutely dependent on environmental fluctuations. This result challenges the intuitive attitudes that the higher population fitness, the more stability is given to that population. Modeling of a system comprised by symbiotic communities has revealed that horizontal transfer confers a selective advantage not only on the acceptor population (which is up to expectations) but also on the donor population. It has therefore been demonstrated that horizontal transfer can be preserved by selection along evolution even without "selfish genes". Evolutionary Constructor can handle any phenotypic trait that is controlled genetically, epigenetically, etc., which extends the applicability of this tool to various processes of information transduction among populations, provided that these processes resemble horizontal gene transfer. PMID- 18415977 TI - Genetic flows in a structured one-dimensional population: simulation and real data on Baikalian polychaetes M. godlewskii. AB - In this work, we apply population dynamic approach enhanced with simulation of the fate of neutrally evolving "DNA sequences" included into each individual in the computer experiment to the case of the Baikalian endemic polychaetes Manavunkia. These animals inhabit a narrow littoral zone around whole the lake perimeter and are of very limited mobility. Accordingly, the general model was modified by addition of a "geographic barrier" of different isolating power and length of existence. Using this model, we simulated the process of genetic differentiation of groups in this organism taking into account isolation by distance and geographical barriers. Wright's F(st) test was used in order to estimate of genetic flow. Relevant sample sizes were estimated, which would produce most important population parameters of the precision required to describe micro-evolutionary processes in Manayunkia. PMID- 18415978 TI - RNAmodel web server: modeling classic attenuation in bacteria. AB - RNAmodel web server was recently established at the IITP RAS to implement our previously proposed model of the classic attenuation regulation of gene expression in bacteria. This new model is based on the concept of resonant-like interaction between RNA polymerase and hairpins of RNA secondary structure formed during transcription. Our modeling relies on standard Monte Carlo procedures and covers all essential stages of the process, including initiation and elongation of transcription and translation; the deceleration of ribosome on regulatory codons, which depends on the concentration of charged amino acyl-tRNA; the polymerase shifting delay caused by secondary structure folded into the mRNA segment between ribosome and polymerase; and, ultimately, either transcription terminating prematurely or polymerase reaching a region of structural genes (antitermination). By means of Monte Carlo simulation we build a function p(c) which expresses the probability of termination (i.e., an enzyme activity) vs. the concentration of charged amino acyl-tRNA or of amino acid itself, measured in actual or relative units. PMID- 18415979 TI - Translation of both complementary strands might govern early evolution of the genetic code. AB - The updated structural and phylogenetic analyses of tRNA pairs with complementary anticodons provide independent support for our earlier finding, namely that these tRNA pairs concertedly show complementary second bases in the acceptor stem. Two implications immediately follow: first, that a tRNA molecule gained its present, complete, cloverleaf shape via duplication(s) of a shorter precursor. Second, that common ancestry is shared by two major components of the genetic code within the tRNA molecule--the classic code per se embodied in anticodon triplets, and the operational code of aminoacylation embodied primarily in the first three base pairs of the acceptor stems. In this communication we show that it might have been a double, sense-antisense, in-frame translation of the very first protein encoding genes that directed the code's earliest expansion, thus preserving this fundamental dual-complementary link between acceptors and anticodons. Furthermore, the dual complementarity appears to be consistent with two mirror symmetrical modes by which class I and II aminoacyl-tRNA synthetases recognize the cognate tRNAs--from the minor and major groove side of the acceptor stem, respectively. PMID- 18415980 TI - Phylogenetic analysis of the p53 and p63/p73 gene families. AB - Proteins of the relative families p53 and p63/p73 are transcriptional factors that are involved in the signaling pathway in cells. The wide spectrum of their functions includes cell cycle arrest and apoptosis in response to DNA damage. The p53 protein also participates in development of particular tissues during embryogenesis. Thus, it is of high importance to establish the relation between structure, function and evolution of these proteins. In the current computational study, the evolutionary mode of the p63/p73 protein family is investigated. Search for the adaptive branches of the phylogenetic tree and the adaptive codons in the nucleotide sequences was performed using the codem1 program from the PAML package, version 3.14. The results obtained were compared with those of our previous phylogenetic analysis of the p53 protein. Evidence was obtained that the evolutionary history of the p63/p73 proteins has been under positive selection. An attempt is made to associate the current evidence with the previous for positive selection in the p53 family. Recently the G245C substitution has been assumed to result in formation of a novel Zn(2+)-binding site in the p53 protein. The molecular mechanics simulations were performed to estimate energy of zinc binding to its site in two dominant-negative p53 mutants--G245C and R175H--in comparison with the wild-type p53. The results of the estimation provided evidence of the novel Zn(2+)-binding site functionality in G245C mutant form. PMID- 18415981 TI - The evolution of the Hh-signaling pathway genes: a computer-assisted study. AB - Positive selection of genes that comprise signaling cascades and play the paramount role in the development of multicellular organisms is critical to our understanding of the reasons for the evolution of embryonic development. In this work, we analyze the evolution of 9 genes involved in the function of the Hh signaling cascade. We demonstrated that positive selection is a characteristic feature of the protein domains, encoded by gene regions, whose functions are related to the molecular mechanisms of development. We also found that the positive selection of Hh-signaling cascade transcription factors, morphogens, their development-related receptors and intracellular signal transduction factors are related to the divergence of the Bilateria taxonomic types. PMID- 18415982 TI - Separation of cytidine 5'-triphosphate biosynthesized from cytidine 5' monophosphate on ion-exchange resin and HPLC analysis of cytidine compounds. AB - Conditions were studied in the biosynthesis of cytidine 5'-triphosphate (CTP) from cytidine 5'-monophosphate (CMP). A 201 x 7 anion ion-exchange resin was applied for the separation of CTP from CMP. Adsorption isotherm and elution conditions (eluant, eluant concentration, flow rate, sample volume loaded) were investigated. At the same time, a new high-performance liquid chromatography on an anion ion-exchange column WAX-1 with UV detector at 260 nm was developed to measure CMP, cytidine 5'-diphosphate (CDP), and CTP. The retention time for CMP, CDP, and CTP are 0.723, 1.448, and 4.432 min, respectively. This new rapid high performance liquid chromatography (HPLC) method for the analysis of cytidine compounds in biological sample has a wide linear range with high precision and repeatability. PMID- 18415983 TI - Identification of a neutralizing scFv binding to human vascular endothelial growth factor 165 (VEGF165) using a phage display antibody library. AB - Vascular endothelial growth factor (VEGF) is a multifunctional cytokine that plays a major role in angiogenesis. Alternative splicing causes the production of several different isoforms (VEGF121, 145, 165, 183, 189, 206). VEGF is essential for tumor angiogenesis, and several studies have correlated elevated VEGF levels with tumor stage, metastases, and progression. We now report the isolation by phage display of human single-chain antibody fragment (scFv) anti-VEGF165. After four rounds of panning against VEGF165, 40 out of 90 phage clones displayed VEGF165-binding activity. One of the positive clones, designated B8, bound to VEGF165 with relatively high affinity and neutralized VEGF165 bioactivity in vitro. The B8 clone was expressed in the soluble form in Escherichia coli HB2151 and purified by immobilized metal affinity chromatography. The purified scFv recognized VEGF165 with the K(D) of 1.80 x 10(-8) M without cross-reaction to VEGF121. In addition to binding, the purified scFv could does-dependently inhibit VEGF165-induced human umbilical vein-derived endothelial cells proliferation. Together with its fully human mature, B8 scFv may have therapeutic implications in therapy of angiogenesis-dependent diseases. PMID- 18415984 TI - Cytochrome P450 BM-3 evolved by random and saturation mutagenesis as an effective indole-hydroxylating catalyst. AB - Cytochrome P450 BM-3 with the mutations A74G, F87V, and L188Q could catalyze indole to produce indigo and indirubin. To further enhance this capability, site directed and random mutageneses on the monooxygenase domain of P450 BM-3 mutant (A74G/F87V/L188Q; 3X) were performed. The mutant libraries created by error-prone polymerase chain reaction were screened using a colorimetric colony-based method on agar plates followed by a spectroscopic assay involving in absorption of indigo at 670 nm and NADPH at 340 nm in microtiter plate. Three mutants (K434R/3X, E435D/3X, and D168N/ A225V/K440N/3X) exhibited higher hydroxylation activity toward indole in comparison to parent enzyme. Moreover, using saturation site-directed mutagenesis at amino acid positions 168, 225, 434, 435, and 440, two P450 BM-3 variants (D168H/3X, E435T/3X) with an up to sixfold increase in catalytic efficiency (kcat/Km) were identified, and the mutant D168H/3X acquired higher regioselectivity resulting in more indigo (dimerized 3hydroxy-indole) compared to parent mutant (93 vs 72%). PMID- 18415986 TI - Development of a new bioprocess for production of 1,3-propanediol I.: Modeling of glycerol bioconversion to 1,3-propanediol with Klebsiella pneumoniae enzymes. AB - Glycerol is a renewable resource for it is formed as a byproduct during biodiesel production. Because of its large volume production, it seems to be a good idea to develop a technology that converts this waste into products of high value, for example, to 1,3-propanediol (1,3-PD). We suggested an enzymatic bioconversion in a membrane reactor in which the NAD coenzyme can be regenerated, and three key enzymes are retained by a 10-kDa ultrafilter membrane. Unfortunately, some byproducts also formed during successful glycerol to 1,3-PD bioconversion runs, as we used crude enzyme solution of Klebsiella pneumoniae. To study the possibilities to avoid this byproduct formation, we built a mathematical description of this system. The model was also used for simulation bioconversions of high glycerol concentration with and without elimination of byproduct formation and of continuous operation. PMID- 18415985 TI - Calreticulin transacetylase mediates the acetylation of nitric oxide synthase by polyphenolic acetate. AB - Our earlier investigations identified acetoxy drug: protein transacetylase (TAase), a unique enzyme in the endoplasmic reticulum (ER) catalyzing the transfer of acetyl groups from polyphenolic acetates (PA) to certain functional proteins. Recently we have established the identity of TAase with ER protein calreticulin (CR) and subsequently transacetylase function of CR was termed calreticulin transacetylase (CRTAase). CRTAase was purified and characterized from human placenta. CRTAase catalyzed the acetylation of a receptor protein nNOS, by a model PA 7, 8-diacetoxy-4-methylcoumarin (DAMC), which was visually confirmed by using antiacetyl lysine. The aim of this report was to provide tacit proof by providing mass spectrometry evidence for CRTAase catalyzed acetylation of purified nNOS by DAMC. For this purpose, purified nNOS was incubated with DAMC and CRTAase, the modified nNOS was analyzed by nanoscale LC-MS/MS, which recorded 11 distinct peptides with a significant score as acetylated on lysine residues. The distribution was in order: lysines-24, -33, -38, -131, and -229 of the PDZ domain, Lys-245 of the oxygenase domain, Lys-754 and -856 of FMN binding domain, Lys-989 of connecting domain and Lys-1300, -1321, and -1371 of the NADPH-binding domain were acetylated. The results documented in this paper highlighted for the first time modification of nNOS by way of acetylation. Our earlier work recorded the profound activation of platelet NADPH cytochrome P-450 reductase and the acetylation of the reductase protein by DAMC, which also remarkably enhanced intracellular levels of nitric oxide. The results reported here coupled with the aforementioned previous observations strongly implicate the possible role of the acetylation of the reductase domain of nitric oxide synthase (NOS) in the NOS activation. In addition, the acetylation of nNOS can be expected to potentiate the interaction with CR, eventually leading to the augmented catalytic activity of NOS and expression of the related biological effects. PMID- 18415987 TI - Corn fiber: structure, composition, and response to enzymes for fermentable sugars and coproducts. AB - Corn (Zea mays L.) fiber, which is the seed coat and residual endosperm left after grain processing, is a low-value residue that contains carbohydrates and aromatic compounds that could provide value-added coproducts. Treatment of corn fiber with NaOH and assessment by gas chromatography indicated a prevalence of ferulic acid, with about 90% ester-linked in the cell walls. p-coumaric acid was much lower at about 10% of the amount of ferulic acid. Histochemical reactions employing acid phloroglucinol and diazotized sulfanilic acid indicated the presence of phenolic acids in cell walls of the pericarp and aleurone layer. Various protocols were tested using milled corn fiber and pretreatment with commercial ferulic acid esterases before cellulase treatment, and dry weight loss and sugars and phenolic acids released into the filtrate were evaluated. Ferulic acid esterases effectively degraded corn fiber and released substantial amounts of ferulic acid and sugars (e.g., glucose and xylose) in the incubation medium. Light microscopy showed that ferulic acid esterase substantially disrupted the aleurone layer but caused little visible damage to the lignified pericarp cell walls. Amounts of compounds released varied with protocols, and one study with various milling methods showed that esterase pretreatment followed by cellulase released about 2.8 to 4.4 and 1.5 to 2.9 times more ferulic acid and glucose, respectively, than cellulase alone. The highest levels for one lot of corn fiber with esterase pretreatment followed by cellulase were 3.9 and 218 mg/g of ferulic acid and glucose, respectively. PMID- 18415988 TI - Aqueous ammonia soaking of switchgrass followed by simultaneous saccharification and fermentation. AB - Simultaneous saccharification and fermentation (SSF) of switchgrass was performed following aqueous ammonia pretreatment. Switchgrass was soaked in aqueous ammonium hydroxide (30%) with different liquid-solid ratios (5 and 10 ml/g) for either 5 or 10 days. The pretreatment was carried out at atmospheric conditions without agitation. A 40-50% delignification (Klason lignin basis) was achieved, whereas cellulose content remained unchanged and hemicellulose content decreased by approximately 50%. The Sacccharomyces cerevisiae (D5A)-mediated SSF of ammonia treated switchgrass was investigated at two glucan loadings (3 and 6%) and three enzyme loadings (26, 38.5, and 77 FPU/g cellulose), using Spezyme CP. The percentage of maximum theoretical ethanol yield achieved was 72. Liquid-solid ratio and steeping time affected lignin removal slightly, but did not cause a significant change in overall ethanol conversion yields at sufficiently high enzyme loadings. These results suggest that ammonia steeping may be an effective method of pretreatment for lignocellulosic feedstocks. PMID- 18415989 TI - Enhanced L-(+)-lactic acid production by an adapted strain of Rhizopus oryzae using corncob hydrolysate. AB - Corncob is an economic feedstock and more than 20 million tons of corncobs are produced annually in China. Abundant xylose can be potentially converted from the large amount of hemicellulosic materials in corncobs, which makes the crop residue an attractive alternative substrate for a value-added production of a variety of bioproducts. Lactic acid can be used as a precursor for poly-lactic acid production. Although current industrial lactic acid is produced by lactic acid bacteria using enriched medium, production by Rhizopus oryzae is preferred due to its exclusive formation of the L-isomer and a simple nutrition requirement by the fungus. Production of L-(+)-lactic acid by R. oryzae using xylose has been reported; however, its yield and conversion rate are poor compared with that of using glucose. In this study, we report an adapted R. oryzae strain HZS6 that significantly improved efficiency of substrate utilization and enhanced production of L-(+)-lactic acid from corncob hydrolysate. It increased L-(+) lactic acid final concentration, yield, and volumetric productivity more than twofold compared with its parental strain. The optimized growth and fermentation conditions for Strain HZS6 were defined. PMID- 18415991 TI - 'Overburdened' staff put mothers and babies at risk. PMID- 18415990 TI - Eleventh-hour changes to GP contract left PCTs with bill. PMID- 18415992 TI - Department plays down medic supply fears. PMID- 18415993 TI - New super-regulator begins to take shape. PMID- 18415994 TI - On local pay and national prices. PMID- 18415995 TI - 'Twas ever thus: why Darzi is 90 years too late. PMID- 18415996 TI - Data briefing. How to spend less while doing more. PMID- 18415997 TI - What a carve-up. PMID- 18415998 TI - Press complaints. School for scandal. PMID- 18415999 TI - Coaching. Learning to be unloved. PMID- 18416000 TI - Ocupational health. Nurse, heal thyself. PMID- 18416001 TI - Professor David Walliker. PMID- 18416002 TI - [Mansonella perstans filariasis]. AB - Mansonella perstans filariasis is widely present in Africa and equatorial America and its pathogenicity has been recently reconsidered. Although M. perstans infection has been considered a minor filariasis, remaining asymptomatic in most of infected subjects, more recent studies have shown that M. perstans is capable of inducing a variety of clinical features, including angioedemas, swellings like the "Calabar swellings" of loiasis, pruritus, fever, headache, pain in bursae and/or joint synovia, or in serous cavities. It is likely that some of the pathological changes observed are induced by the immune response to the infection. Eosinophilia is present in many cases of infection. Moreover M. perstans filariasis is difficult to be treated. Effective treatment is lacking and there is no consensus on optimal therapeutic approach. The most commonly used drug is diethylcarbamazine (DEC) that is however often ineffective. Although other drugs have been tried (e.g. praziquantel, ivermectin), none has proven to be reliably and rapidly effective. Mebendazole seemed more active than DEC in eliminating the infection, with a comparable rate of overall responses. Thiabendazole evidenced a small, but significant activity against the infection. Combination treatments (DEC plus mebendazole) resulted in a significantly higher activity compared with the single drugs. PMID- 18416003 TI - Aedes albopictus in Sardinia: reappearance or widespread colonization? AB - The Asian tiger mosquito Aedes albopictus (Skuse, 1894) was first discovered in the South of Sardinia in October 1994, in a tyre depot not far from Cagliari Elmas airport. Insecticide treatment was thought to have successfully eradicated the mosquito, but in 1996 and 1997 new breeding sites were discovered, a few at some distance from the first. More recently two sites have been reported in the heart of the city of Cagliari. It is not known whether the mosquito has spread from the first breeding place discovered, where treatment may not have been definitive, or whether they have been newly introduced. The recent sighting of Ae. albopictus in Olbia in the Northeast of the island tends to suggest the latter. Cagliari and Olbia are actually Sardinia's two largest sea ports of entry. PMID- 18416005 TI - AIDS vaccine efforts move forward, but full prevention is elusive goal. Experts talk about what is possible soon. PMID- 18416004 TI - Diagnosis of congenital toxoplasmosis: pre- and post-natal evaluation in Sicilian (Italy) epidemiological area. Preliminary data. AB - To evaluate the usefulness of conventional serological methods with western blot assay (WB) in congenital toxoplasmosis diagnosis, we prospectively enrolled in a clinical and serological follow-up all pregnant women with Toxoplasma gondii infection and their offspring, referred to us from October 2004. Western blot and standard serological test were performed on sera collected from mother during pregnancy and from mother and child at birth, at postpartum month 1-3-6-9 and 12. At this point in time, 22 pregnant women and 14 infants have completed the follow up. 4 newborns were infected and 2 had specific toxoplasmosis anomalies at the birth. In mothers without seroconversion, the WB performed during pregnancy demonstrates the highest accordance with postnatal follow-up whereas in 1 case the negative result of PCR analysis was not confirmed by postnatal observation. The detection of anti-T gondii IgG against 8 kDa accessory antigenic band and against the accessory band included between 35 and 40 kDa band in immunoblot assay was useful for diagnosis of acute phase but did not improve the evaluation of comparative postnatal profile. Althougth few infants have concluded the postnatal follow-up, the preliminary results showed a greater value of using a IgM and IgA WB test than other standard method for the early diagnosis of toxoplasmosis at birth also in child born to treated mothers. The comparative anti-T gondii IgG immunoblot profile of mother and child permitted us to reduce the time of ruling out infection in newborns born to mothers with probable or possible infection and/or when prenatal diagnosis is negative or not performed. PMID- 18416006 TI - New recommendations out on HIV & circumcision. PMID- 18416007 TI - Researchers make strides in global battle against HIV. Scientific advances may aid in stemming spread of HIV. PMID- 18416008 TI - Researchers note changing patterns of HIV morbidity. Study suggests benefit to starting ART early. PMID- 18416009 TI - FDA notifications. Tentative approval granted for combo antiretroviral drug. PMID- 18416010 TI - FDA notifications. Abbott sends out provider letter about Kaletra for kids. PMID- 18416011 TI - FDA notifications. Tentative approval for generic lamivudine/zidovudine. PMID- 18416012 TI - Drop-off in mammograms may help explain breast cancer decline. PMID- 18416013 TI - Obstructive sleep apnea. More than just a snoring problem. PMID- 18416014 TI - Mayo Clinic office visit. MRI and breast cancer screening. An interview with Stephen Phillips, M.D. PMID- 18416015 TI - How to relax. Making the most of your downtime. PMID- 18416016 TI - I've heard that I should look for the amount of elemental calcium when choosing a calcium supplement. What is elemental calcium, and how can I tell how much a supplement contains? PMID- 18416017 TI - Experience in control of avian influenza in the Americas. AB - The outbreaks of highly pathogenic avian influenza (HPAI) in Canada in 1966, the USA in 1984 and Mexico in 1994 led to a clear increase in biosecurity measures and improved intensive poultry production systems. In the past 12 years (1994 2006), there have been four outbreaks of HPAI on the American continent: in Mexico in 1994 (H5N2), in Chile in 2002 (H7N3), in the USA in 2004 (H5N2) and in Canada in 2004 (H7N3). In all cases, the control and eradication measures were based on prompt detection, depopulation of infected farms and epidemiological contacts, increased biosecurity measures and control of the movement of live poultry and their products, by-products and infected material. In Mexico, in addition to the aforementioned measures, the use of massive vaccination allowed eradication of HPAI in a relatively short time in two affected areas of high density commercial poultry. PMID- 18416018 TI - The pediatric electrocardiogram: part I: Age-related interpretation. AB - Emergency physicians attending to pediatric patients in acute care settings use electrocardiograms (ECGs) for a variety of reasons, including syncope, chest pain, ingestion, suspected dysrhythmias, and as part of the initial evaluation of suspected congenital heart disease. Thus, it is important for emergency and acute care providers to be familiar with the normal pediatric ECG in addition to common ECG abnormalities seen in the pediatric population. The purpose of this 3-part review will be to review (1) age-related changes in the pediatric ECG, (2) common arrhythmias encountered in the pediatric population, and (3) ECG indicators of structural and congenital heart disease in the pediatric population. PMID- 18416019 TI - Correction to the PRoFESS Trial paper by Diener et al. (Cerebrovasc Dis 2007;23:368-80. PMID- 18416020 TI - Methods in molecular biology. Preface. PMID- 18416021 TI - What is acceptance? PMID- 18416022 TI - Lung cancer: beyond the guidelines. PMID- 18416023 TI - Should cost of care be considered in a clinical practice guideline? PMID- 18416025 TI - Introduction to the special section on BioGrid: biomedical computations on the grid. PMID- 18416026 TI - [Quality of life evaluation in residential facilities for the mentally ill in Puglia (Italy). Phase 2: evaluation of quality of care]. PMID- 18416027 TI - [Health literacy: the new outcome indicator for evaluating a health education program]. AB - Through health promotion campaigns, health literacy has become a vital factor in determining a population's health in terms of the macro view of health. Since the 1990s, the concept has been increasingly valued in American health circles. Health literacy not only means understanding medical information, but also implies the degree to which individuals have the capacity to obtain, process, and understand the basic health information and services needed to make appropriate health decisions. There are three instruments for estimating individuals' health literacy, including the Rapid Estimate of Adult Literacy in Medicine (REALM) , the Short-Test of Functional Health Literacy in Adults (Short-TOFHLA) and the National Assessment of Adult Literacy (NAAL). Literature review reveals that health literacy is an important indicator for evaluating the effectiveness of a health education program for the 21st century. The author suggests that nurses adopt such strategies for health education as designing educational materials that prompt introspection, using a range of Internet technologies and designing strategies of community empowerment to conduct health education programs, in order to improve clients' health literacy and raise the effectiveness of such programs. PMID- 18416028 TI - Oncoproteins, heterochromatin silencing and microRNAs: a new link for leukemogenesis. AB - The pathogenesis of acute myeloid leukemias involves complex molecular events triggered by diverse alterations of genomic DNA. A limited number of initiating lesions, such as chromosomal translocations generating fusion genes, are constantly identified in specific forms of leukemia and are critical to leukemogenesis. Leukemia fusion proteins derived from chromosomal translocations can mediate epigenetic silencing of gene expression. Epigenetic deregulation of the DNA methylation status and of the chromatin "histone code" at specific gene sites cooperate in the pathogenesis of leukemias. The neutralization of these crucial oncogenic events can revert the leukemia phenotype. Thus, their identification and the study of their molecular and biological consequences is essential for the development of novel and specific therapeutic strategies. In this context, we recently reported a link between the differentiation block of leukemia and the epigenetic silencing of the microRNA-223 gene by the AML1/ETO oncoprotein, the product of the t(8;21) the commonest AML-associated chromosomal translocation. This finding indicates microRNAs as additional epigenetic targets for leukemogenesis and for therapeutic intervention in leukemias. PMID- 18416030 TI - Medicare finances: findings of the 2008 Trustees Report. AB - Each year Medicare's Board of Trustees issues a report that describes the financial condition of the Medicare program in the near term and over the next 75 years. Similar to recent trustees reports, the 2008 report projects that Medicare's Hospital Insurance Trust Fund will be depleted in 2019, at which time scheduled income will cover 78 percent of expenditures. Looking at these projections, some observers contend that Medicare must be fundamentally restructured in order to put the program--and the overall federal budget--on a sound fiscal course. Others point out that Medicare spending has grown at about the same rate as spending for private health insurance and argue that it is impossible to limit spending on Medicare without also slowing the growth of private health care costs or abandoning equal access to care for the aged and disabled. PMID- 18416029 TI - Epigenetic mechanisms of gene regulation during mammalian spermatogenesis. AB - The process of spermatogenesis is a unique form of cellular differentiation and is regulated by genetic and epigenetic factors. Recent studies have shown that some epigenetic factors (histone variants and histone modifying enzymes) are exclusively expressed during spermatogenesis; the disruption of these factors leads to spermatogenic defects. Also, during spermatogenesis a key epigenetic process termed meiotic sex chromosomes inactivation (MSCI) occurs; MSCI leads to the inactivation of most genes on the X and Y chromosomes; however, the mechanism of MSCI is distinct from X inactivation in female somatic cells. Furthermore, a new class of non-coding RNAs (i.e., piRNAs) has recently been identified which is exclusively expressed during spermatogenesis. This review discusses recent advances in our understanding of epigenetic mechanisms that operate during spermatogenesis with specific emphasis on histone modifications, MSCI and non coding RNAs. Finally, we propose that spermatogenesis can be a powerful experimental system to dissect epigenetic mechanisms of gene regulation. PMID- 18416031 TI - [Historical data and some comments on closed techniques in cardiac surgery]. AB - On the 29th of May, 1907 Ortiz de la Torre preformed, in the General Hospital of Madrid, the first surgery to treat a cardiac wound. In this same year Rehn reported the first 124 cases of sutured cardiac wounds, following the first one, performed by him, in the world on the 9th of September, 1896 which opened the cardiac surgery. This started with the so called closed techniques. We are presenting some data according to our own experience in honor to those pioneers surgeons. The most important contributions are reviewed as follows: 1. The suture of the heart's wounds with the definitive aportation of Rehn on the 9th of September, 1896. 2. The extracardiac surgery related with: a) Pathology of the pericardium with the proposals of Weill in 1895 and Delorme in 1898, about removing it in case of constrictive pericarditis and the first decortication executed by Rehn in 1920. The first case of removing a pericardium's cyst by Pickhardt in 1934, is mentioned. b) The great vessels' pathology with the contribution of Trendelenburg (1908) in case of lung embolie and the first patient operated with success by Kirschner in 1924. The ligature of the patent ductus arteriosus done by Gross and Hubbard in 1939, with the previous findings of Munro and Strieder. The resection of the narrow segment in coarctation of the aorta done by Crafoord, under sugerence of Nylin, in 1944. c) Extracardiac surgery in order to correct intracardiac pathology with reference to the several types of systemic-pulmonary anastomosis, starting with Blalock-Taussig operation in Fallot's tetralogy done the first time November 29th of 1944, and after that the Pott's aorto-pulmonary anastomosis (1946), the superior vena cava-right pulmonary artery anastomosis or Glenn's operation (1954), the Waterston's ascending aorta-right pulmonary artery anastomosis (1962) with the Cooley's modification (1965). It is also mentioned the banding of the pulmonary artery after Miller and Dammann (1952) as well as the techniqes aiming to correct the pulmonary hypertension in mitral stenosis. Lastly he quotes venous transposition operations in the treatment of transposition of the great vessels (Lillehei, Greech, Baffes). 3. With regard to the group of closed techniques which aim is to correct intracardiac pathology, the following techniques are taken into account: Blalock-Hanlon's operation (1948) that create an interauricular communication in case of transposition of great arteries and Rahskind's technique. Within mitral stenosis, sugerence of sir Lauden Brunton (1902), efforts of Cutler, Levine and Beck and the pioneer Souttar's surgical procedure, in 1925, are described. Afterwards Bailey's intervention (10th Juny 1948) and Harken and other surgeons are mentioned. Figuera's instrument, introduced through left auricle is also described. The first efforts to correct aortic and pulmonary stenosis are described from the initial contributions of Tuffier, Doyen (1913), Brock and Sellors (1948) ending with the ingenious attempts of closing interauricular communication with Swan's and Hufnagel's buttons (1950), Bailey's atrioseptopexy, Sondergaard's technique and Gross'well technique. Finally the pioneer Bailey's aportation (February 6th 1951) for closing interventricular septal defect using a tube of pericardium introduced through the interventricular defect, going out through the wall of the left ventricle. PMID- 18416032 TI - [Microbes: pathogen or no pathogen?]. AB - The pathogenic activity of a microorganism in a host depends on its genome, of the environmental factors that it is and of the immunity of the host. The recent advances are studied in the first two factors, so much the denominated "pathogenicity islands", that condition the virulence, as the external factors (temperature, pH, concentrations of metals, etc.) that they allow or not the development of this pathogenic capacity. The new genetics knowledge and molecular microbiology will allow important advances in the pathogenesis, diagnosis, treatment, epidemiology and prevention of the illness caused by biological agents. PMID- 18416033 TI - [HIV-1 genotypic resistance]. AB - Three sections have been studied: the characteristics of the virus (to insist on in their genome, able to produce structural and regulating proteins), and mainly, their biological cycle that allows us to know where the different antiretroviral will act; the resistance phenomena produced by their high answered and mutant capacity; and on third place, it show up some results of the investigations carried out in the Hospital Universitario San Cecilio, Granada, on two aspects: realization and validation of algorithms of interpretation of mutation based on virologic answer, and comparison of the influence according to the adult or smaller number of mutations appeared with the resistances in front of active medicines. PMID- 18416034 TI - [Functional digestive disorders and fibromyalgia]. AB - Fibromyalgia is a syndrome characterized by generalized chronic pain, with mainly musculoeskeletal and excess tenderness in certain areas of the body of unknown origin. Patients with fibromyalgia are often affected by additional pain symptoms, mostly funtional disorders. People with fibromyalgia experience migraines as well as symptoms of irritable bowel syndrome (IBS). Others digestive functional disorders have not been yet associated with fibromyalgia. We enclosed preliminary results of a study including all functional disorders according to Rome II criteria. We conclude that these disorders are more prevalent in patients with fibromyalgia and not only the IBS. These functional disorders are also associated with some emotional factors. All these factors make that quality of life in those patients were not good. PMID- 18416035 TI - [Perspectives on Benito Perez Galdos' cataracts]. AB - Galdos started with sight troubles before he was 40 years old. Later, he suffered an iritis and he had more marked cataract in his left eye. When he was 62 years old he suffered a transitory hemiplegia. In 1911 and in 1912 he was operated on for cataracts by the Dr. M. Marquez. At the same time, he presented joint pain, lightning pain especially in their lower extremities, ataxic gait, Romberg's sign, complete blindness and the impossibility of standing up, as a result he had to stay in the bed until his death. I think that Galdos suffered from tertiary syphilis (late) manifested by tabetic neurolues and ocular syphilis, which was the cause of his blindness, and besides arterioloesclerosis and hypertension. PMID- 18416036 TI - [Pathochrony and presage of obesity]. AB - The obesity is the metabolic most frequent illness; it affects at least to the population's of adults 12% and it produces increase of the morbimortality. We analyze their diverse mechanism: endogenous, constitutional, hyperbulimic, for sedentary, family or hereditary, senile, hypothalamic, neuro-psychic, endocrine metabolic, characteristic of certain races or ethnos, for alterations of the trofic appetite center, syndrome of Morgagni-Stewart-Morel, tumors of the islets of Langerhans, etc. Also, we index therapeutic rules. PMID- 18416037 TI - [The climate change view for a physician]. AB - There is no doubt that a climate change is happening with an increase of the mean temperature of the planet and an ensemble of extreme geologic and climatic phenomena. The causes are not well known and they seem produced by gases generated by fossil combustibles. As physicians we are interested in foreseeing solutions to the unknown repercussions of this change in the biosphere and the human life. Most of the world governments are dedicating resources to study this problem. The reduction of the use of this combustibles and the increase of the forest mass seem the most adequate solutions. PMID- 18416038 TI - [Glucolipotoxicity, resistance to the action of insulin and type 2 diabetes mellitus]. AB - At present the diabetes mellitus (DM) constitutes a tremendous sanitary problem with values of morbility, prevalence and economical costs highly elevated, mainly because type 2 DM. These facts have stimulated the scientific research on the processes of glucolipotoxicity and of insulin resistance and their contribution to the development of type 2 DM. Both hyperglycemia and hyperlipidemia are important biochemical alterations in the DM that produce toxic effects. Chronic hyperglycemia predisposes to the micro and macrovascular complications of DM through the glucoadaptation, glucotoxicity and glucoapoptosis steps, in which the alterations of the hexosamine pathway, glycation of key proteins on the insulin action cascade and oxidative stress of beta-cells produce a decrease in the synthesis and secretion of insulin together with resistance to this hormone. Also, the lipotoxic process is preceded by a lipoadaptation period characterized by the induction of genic programs directed to eliminate the excess of lipid metabolites through the activation of lipolitic routes and inhibition of lipogenesis. Nevertheless the persistence of hyperlipidemia produced a lipotoxicity facilitates the apparition of lipoapoptosis in which ceramides and nitric oxide play a role. Glucolipotoxicity concept was proposed by Prentki and Corkey in order to explain the metabolic alterations present in type 2 DM associated to obesity, such as increased abdominal fat, insulin resistance and abnormal insulin secretion. It is known that insulin resistance play an important role in the development of type 2 DM, until the point that it is already present 10 to 20 years before type 2 DM starts and it is a good predictor of this nosologic entity. Accordingly, a better knowledge of the mechanisms implied on insulin resistance may be very useful for the future treatment of patients with type 2 DM, which is considered as one of the great epidemics of the XXI century. PMID- 18416039 TI - [Promotion of basic research in the European Union: the European research council (ERC) and the initiative for science in Europe (ISE )]. AB - Europe has today a function of intellectual guidance and watch tower that will be unable to accomplish if remains far of the forefront in scientific research and application of knowledge The "delocalization of talents" towards the west is much worst than that of production towards the east. From January 1st 2007, the ERC, with an annual budget of 1.5 billion Euros, represents an important step to improve the situation. The ISE, representing the European scientific community, will further enhance it. PMID- 18416040 TI - [In memory of his excellency Mr. D. Antonio Fernandez de Molina]. PMID- 18416041 TI - [Obesity and not alcoholic esteatohepatits]. PMID- 18416042 TI - [Health and development, achievements and contrasts]. PMID- 18416043 TI - [Effect of L-arginine, vitamins C and E, and omega-3 acids (DHA, EPA) on oxidative stress and endothelial dysfunction in the mouse model of renal insufficiency]. PMID- 18416045 TI - While most diabetes drugs provide similar glucose control, some offer important advantages. PMID- 18416044 TI - New report on decreasing asthmagens in hospitals. PMID- 18416046 TI - New trends in foreign nurse recruitment. Interview by Diane E Scott. PMID- 18416047 TI - Were 'striking nurses' eligible for unemployment benefits? PMID- 18416048 TI - Home health nurse botches care of pressure ulcers. Case on point: Olsten Health Services, Inc. v. Cody, 2008 FLCA3 3D07-212- (03/05/08) So.2d -FL. PMID- 18416049 TI - AR: Forensic RN testified in child rape case: testimony allowed in evidence failure to object. PMID- 18416050 TI - Nurse sued employer & dr. for assault/false imprisonment by dr. Case on point: Baker v. Cook Children's Physician Net., 2008 TCA2 2-07-174 (0228/08) S.W.3d -TX. PMID- 18416051 TI - [ZAP-70--a marker in chronic B-cell lymphocytic leukemia]. PMID- 18416052 TI - [Clinical, morphologic and immunohistochemical factors of prognosis in papillary cancer of the thyroid]. PMID- 18416053 TI - [Gail coefficient as a risk factor and prognosticator for the course of breast cancer: correlation with hormonal and metabolic parameters and clinical and morphologic features]. AB - Gail coefficient (GC) generally used in breast cancer predictions for the next 5 year--or entire survival was determined in both patients and healthy controls of the same age, residents of St. Petersburg. Simultaneously, a correlation was established with hormono-metabolic indices, receptor pattern, tumor stage and size and some other characteristics. GC in cancer patients with age <50 was significantly higher than in control. In menopausal cancer patients, greater GC correlated with such parameters as body mass, weight index, glucose, total cholesterol and low density lipoproteids after fasting. The latter group showed a tendency towards enhanced estradiol and testosterone in blood serum. In reproductive patients with elevated GC, estradiol level rise was significantly lower and most tumors were receptor-negative. However, involvement of regional nodes was relatively rare. To summarize, GC determination characterizes risk and certain clinico-morphological features of distinction between reproductive and menopausal patients. PMID- 18416054 TI - [Frequency of 5382insC mutation of the BRCA1 gene]. AB - The paper assesses c5266dupC (5382insC) mutation incidence among breast cancer patients, residents of the Republic of Bashkorstan and Tyumen Region. It appeared as high as 4%. PMID- 18416055 TI - [Prediction of relapse-free period after radio- and chemotherapy in patients with breast cancer]. AB - The study was carried out in 55 females with breast cancer stage III-IV, aged 35 77 yrs. The patients received radiation and standard courses of CMF, or CAF, or CMF+trastuzumab, or carboplatin+vepesid, or taxotere+epirubicin, or mustoforan. Radiation was administered in fractionated single focal doses of 3 Gy. S-index was determined before treatment started by an original short (ca. 4 hrs) method of evaluating apoptotic leukocyte DNA disintegration after ex vivo test irradiation as described by Ivanov S.D. et al., 2003. The index ranged 0.40-1.44 rel. units thus correlating with 2-100 month relapse-free period. Direct correlations were identified between S-index and relapse-free period for different regimens of radiochemotherapy. S-index level in excess of 0.95 involved relatively longer relapse-free survival. In summary, our method offers means of predicting the effectiveness of a particular regimen prior to treatment thus making individualization possible. PMID- 18416056 TI - [Prognostic significance of cyclooxygenase (COX-1 and COX-2) expression in endometrial carcinoma (clinical and immunohistochemical study)]. AB - An immunohistochemical study of 100 endometrial adenocarcinomas assessed expression of COX-1 (40%) and COX-2 (72%) cyclooxygenases. An inverse correlation between COX-2 expression and tumor cell differentiation was matched by a direct one between COX-2 expression and survival. In cases of no expression, overall relapse-free 5-year survival was 92 and 88%, respectively, while distinct expression was associated with 52 and 48%, respectively (p = 0.0004; 0.0005). Similar relationships were reported for COX-1, although the significance was lower (p = 0.02). Since the differences in survival vis-a-vis clinico morphological features were insignificant, except for COX-1, COX-2 expression, the latter characteristics should be regarded as independent prognosticators, much more superior to such traditional factors as stage and cell differentiation. PMID- 18416057 TI - [Molecular, genetic and immunohistochemical investigation of carcinosarcoma of the female reproductive tract]. AB - Tissue precursors and genesis of female reproductive tract carcinoma vis-a-vis its carcinomatous and sarcomatous patterns remain unknown. To determine the clonal origin of 17 female reproductive tract carcinomas, such molecular, genetic and immunohistochemical techniques as PCR-SSCP and/or denaturing gel electrophoresis for K-ras, p53 and PTEN genes; D17S786, CHRNB1, TP53, BAT26 and BAT40 microsatellites and immunostaining for p53 protein were used. Carcinomatous and sarcomatous components were studied separately. Eight tumors were assumed to be monoclonal (combination or conversion tumors), while one--of an obscure origin. Our results suggest that carcinosarcomas were characterized by chromosomal instability. Moreover, it was shown that it is necessary to combine immunohistochemical techniques with a battery of methods including genetic ones to determine clonal origin of immunologically--stained carcinosarcomas. PMID- 18416059 TI - [Study of lactate dehydrogenase isoenzyme in the cerebrospinal fluid of patients with acute lymphoblastic leukemia]. AB - Lactate dehydrogenase isoenzymes were profiled in cerebrospinal fluid in patients with acute lymphoblastic leukemia. Cytological examination of patients with central nervous involvement identified isoenzyme 5 which failed to show after selective treatment. The isoenzyme's detection in cerebrospinal fluid without typical cytological characteristics may be regarded as a precursor of such complication. Hence, this may be used as an additional criterion for refinement of cytological evidence employed in diagnosis and prognosis of central nervous involvement in patients with acute lymphoblastic leukemia. PMID- 18416058 TI - [A comparison of two risk-adapted regimens of therapy for Hodgkin's disease in children and adolescents]. AB - The paper discusses the advantages offered by two regimens of therapy of Hodgkin's disease in children and adolescents (139)--DAL-HD (versions 87 and 90) (83) and SPbHD-05 (56). Survival rates were fairly high (OS--94.3%; DFS-- 90.0% and EFS--81.2%); they differed depending on risk group. Both the potential of a largen choice of prognostic criteria used in risk grading and the hazards from akylating drugs and anthracyclines were demonstrated. PMID- 18416060 TI - [Specific features of endogenous autorosette formation in the peripheral blood of cancer patients]. AB - The nature and rate of endogenous autorosette formation were investigated in peripheral blood from cancer patients. Both neutrophils, monocytes and platelets were involved. The latter lysed red blood cells. Thrombotic autorosette levels rose as the number of lymphocytes was falling and metastases appeared. PMID- 18416061 TI - [Effectiveness of multislice computed tomography for evaluating vascular invasion in retroperitoneal tumors]. AB - The potential of combined application of multislice computed tomography and angiography for evaluation of vascular invasion in retroperitoneal tumors has been assessed. The predictive value of the method was tested by comparing preoperative CT and angiographic data with intraoperative findings on vascular invasion--abdominal part of the aorta and its large branches, inferior vena cava and common iliac veins--in 44 patients; primary tumor--26, recurrent--18, pelvic localization--7, intraperitoneal--37. Thirty-one combined operations were performed. Multislice scans were obtained by using 4 rows of solid-state detectors. For intravenous injections, non-ionic contrasting substance Omniak (loxegol, Amersham, Ireland), 350 mg I/ml was used. PMID- 18416063 TI - [Use of radiochemotherapy for inoperable non-small cell lung cancer]. AB - The study was concerned with efficacy and tolerability of radiochemotherapy for inoperable non-small cell lung cancer using fractionated single focal dose of radiation in conjunction with vinorelbine or 5-fluorouracil. The treatment proved moderately toxic and was frequently followed by complete regression and survival lasting 12-36 months. Also, it was cost-effective due to shorter courses of radiotherapy. PMID- 18416062 TI - [Effect of light regimens and melatonin on homeostasis, life span and spontaneous tumorigenesis in male rats]. AB - The effect of various light regimens--standard, 12 hrs light:12 hrs darkness, natural lighting in the North-West of Russia and constant illumination--on homeostasis, life span and spontaneous tumorigenesis has been studied in male rats. It was shown that constant exposure or natural lighting involved accelerated aging as evaluated by coefficient of homeostatic stability and survival curves shifting to the left. Constant illumination was followed by a significantly faster spontaneous tumorigenesis as compared with standard lighting. Nocturnal administration of melatonin with drinking water (10 mg/l) prevented the adverse effects of constant and natural lighting on homeostasis and inhibited spontaneous tumorigenesis, particularly, that in the hemopoietic system. PMID- 18416064 TI - [Knowledge of lung cancer patients about their disease and their psychological adaptation]. AB - Among 42 patients with primary lung cancer, 11 refused, 20 admitted while 11 admitted partially the fact of their disease. Later on, compensatory mental dysadaptation in the first group was followed by inadequate attitude to the disease. The growing awareness of the disease involved neurotic symptom enhancement which was an adequate response under the circumstances. On the other hand, adequate self-assessment and finding ways of self-expression promoted psychological adaptation. The better the patient was informed of his or her condition, the more their sense of responsibility for it was stimulated. PMID- 18416065 TI - [Inflammatory breast cancer: clinical course, diagnosis and treatment]. AB - To identify influence of different therapeutic and prognostic factors on survival, a review of the data on treatment of 197 patients with inflammatory breast cancer was carried out at the Dispensary (1994-2004). Multimodal treatment included combinations of radiotherapy, mastectomy and chemotherapy (CMF--75; CAF- 98 and taxanes--9). Significantly lower survival was reported for patients with lymph node involvement (N1, N2 and N3) and those without mastectomy and radiotherapy. Overall 10-year survival was registered only in those who had received chemotherapy, radiotherapy and mastectomy (20.2% +/- 5.5); overall 5 year survival was 42.0% +/- 4.8. PMID- 18416067 TI - [Combination therapy of localized and locally-advanced renal-cell cancer using preoperative arterial embolization]. PMID- 18416066 TI - [Transcatheter treatment of liver metastases from breast cancer]. AB - Transcatheter treatment for liver metastases from breast cancer was given to 55 patients (1995-2006): hepatic artery infusion (HAI) with 5-fluorouracil, doxorubicin and carboplatin--11; hepatic artery oil chemoembolization (HAOC) with doxorubicin--14; taxotere (Docetaxel)--17; HAI+HAOC--13. Partial response was reported in 18%, stabilization--27%, metastatic progression--55%. Mean survival (MS) of all deceased patients--11.9 +/- 1.5; 1-, 2- and 3-year survival--40, 20 and 0%, respectively. HAOC: partial response and stabilization--42%, progression- 58%. As of January 2007, 7 patients survived 8-22 months (mean 14.3 months); 24 deceased had survived 16.8 +/- 3.3 months. HAOC+HAI: complete response--8%, partial--23%; stabilization--44%; progression--25%. Three patients have survived 24, 33 and 82 months; mean survival of 10 deceased--19.9 +/- 4.5 months, their 1 , 2- and 3-year survival was 90, 60 and 20%, respectively.. Mean survival of doxorubicin-treated patients was 18.9 +/- 1.6, taxotere--24.8 +/- 5.1 months (p < 0.05); 1-, 2- and 3-year survival- 57, 29 and 7%, and 88, 35 and 24%, respectively. HAOC+HAI was the most effective while application of taxotere was followed by longest mean survival. PMID- 18416068 TI - [Assessment of mucosal injury in stage T1-T2NOMO bladder cancer after radiation or intravesical chemotherapy]. AB - The report reviews the experience gained with radio- and chemotherapy-related injuries suffered by bladder cancer patients. It corroborates the opinion of most European specialists that indications for radical cystectomy be extended with due considerations of up-to-date potential in anesthesiology, extensive care and pharmacology. Possible untoward side effects of radiation and intravesical chemotherapy as well as means of solution of the problem are discussed. It is urgent considering the constantly growing number of such patients, their age and associated somatic problems. PMID- 18416069 TI - [Accelerated postoperative chemoradiotherapy for malignant cerebral glioma]. AB - Procedure of accelerated immunotherapy for cerebral glioma is presented. Large fractions of radiation ranged from 3 Gy, 5 times a week, to a total focal dose of 51 Gy. After accumulation of total doses of 18, 33 and 48 Gy, vincristine was injected intravenously. Urea derivatives were given on reaching 21, 36 and 51 Gy. Treatment with the immunomodulator roncoleukine was carried out on completion of radiotherapy. An evaluation of the immediate end results of accelerated immunotherapy showed improved survival as compared with standard treatment. PMID- 18416070 TI - [High-intensity ultrasonic destruction of fatty tissue in the course of lymphatic dissection in oncological patients]. AB - The paper discusses experimental findings in support of ultrasound destruction of fat tissue at different sites. Rate of destruction was designed to match that of development of the connective-tissue stroma of adipose tissue and size of adipose cells. Parameters of adipose tissue framework at different sites may be taken into account during extended lymph node dissection using ultrasound liposuction in oncological practice. PMID- 18416071 TI - [Current methods of radical treatment for localized prostate cancer]. PMID- 18416072 TI - The combined use of acupuncture and Chinese medicines for treatment of menopausal syndrome--a clinical report of 63 cases. AB - OBJECTIVE: To evaluate therapeutic effects of the combined use of acupuncture and Chinese medicines in 63 cases of menopausal syndrome. METHODS: Ninety-three cases of menopausal syndrome were randomly divided into 2 groups. Sixty-three cases of the treatment group were treated by the combined use of acupuncture and Chinese medicines. Thirty cases in the control group were only given Western medicines. RESULTS: The total effective rate was 96.83% in the treatment group, and 73.33% in the control group. CONCLUSIONS: The combined use of acupuncture andChinese medicines is more effective for treating menopausal syndrome. PMID- 18416073 TI - Acupuncture treatment for optic nerve contusion. PMID- 18416074 TI - Acupuncture combined with spinal tui na for treatment of primary dysmenorrhea in 30 cases. AB - OBJECTIVE: To observe the therapeutic effects in acupunture treatment of primary dysmenorrhea combined with spinal Tui Na, and study its mechanism. METHODS: Thirty cases of the treatment group were treated by acupuncture combined with spinal Tui Na, and thirty cases in the control group were treated by routine acupuncture. RESULTS: The total effective rate was 93.3% in the treatment group, and 73.3% in the control group, with a significant difference between the two groups (P < 0.05). CONCLUSIONS: Acupuncture combined with spinal Tui Na has good prospects for treatment of primary dysmenorrhea. PMID- 18416075 TI - Insomnia due to deficiency of both the heart and spleen treated by acupuncture moxibustion and Chinese tuina. AB - OBJECTIVE: To observe therapeutic effects of the comprehensive therapy of acupuncture-moxibustion and Chinese Tuina for treatment of insomnia due to deficiency of both the heart and spleen. METHODS: 92 cases were divided randomly into the treatment group (treated by acupuncture-moxibustion and Chinese Tuina) and the control group (treated by acupuncture-moxibustion). RESULTS: The therapeutic effect of the treatment group was obviously superior to that of the control group (the CHI2 test showed P < 0.01). CONCLUSIONS: The comprehensive therapy of acupuncture-moxibustion and Chinese Tuina can give marked therapeutic effects for treatment of insomnia due to deficiency of both the heart and spleen. PMID- 18416076 TI - Electro-acupuncture combined with the trigger point needle-embedding for treatment of primary trigeminal neuralgia in 31 cases. PMID- 18416077 TI - Forty cases of gastrointestinal neurosis treated by acupunture. AB - OBJECTIVE: To compare the therapeutic effect of acupuncture for gastrointestinal neurosis with that of oral remedy. METHODS: Eighty cases were randomly divided into the following 2 groups. In the treatment group, acupuncture was given for one month at the main points of Zhongwan (CV 12), Zusanli (ST 36), Taichong (LR 3) and Shenmen (HT 7), with the auxiliary points selected according to TCM differentiation. In the control group, Domperidone was orally administered for one month. RESULTS: The total effective rate was 92.5% in the treatment group and 75.0% in the control group, with a significant difference between the 2 groups (chi2 = 4.423, P < 0.05). Acupuncture was superior to the oral remedy in therapeutic effects. CONCLUSIONS: Acupuncture may show better results for gastrointestinal neurosis and with less toxic side effects. PMID- 18416078 TI - Clinical observation on the effect of earlobe-bleeding plus body acupuncture in 85 cases of common acne. AB - OBJECTIVE: To observe the clinical effect of earlobe-bleeding plus body acupuncture for common acne. METHODS: The earlobe-bleeding plus local and body acupuncture based on syndrome differentiation was adopted for 85 cases of common acne. RESULTS: Of the 85 cases treated, 61 were cured, 12 markedly effective, 9 improved, and 3 failed. The total effective rate was 96.47%. CONCLUSIONS: The therapy can treat both the Biao-symptom and Ben-root of common acne, short in course and effective in result. PMID- 18416079 TI - External application of herbal medicine to acupoints. AB - Application of herbal medicine to acupoints is to regulate the meridians, yin yang, and qi and blood for preventing and treating diseases through the pharmacological action of herbal medicines and with their stimulation to the acupoints. This article explains how to apply herbal medicines and gave the examples for the treatment of hypertension, asthma, chronic bronchitis and allergic rhinitis. Application of herbal medicines to acupoints is one of the important components of TCM, which shows satisfactory effects in treatment of some chronic diseases. PMID- 18416080 TI - Leptin, ghrelin and TNF-alpha before and after hypo-caloric traditional Chinese diet and auricular acupuncture. PMID- 18416081 TI - Clinical effects of shenqi fuzheng injection in the neoadjuvant chemotherapy for local advanced breast cancer and the effects on T-lymphocyte subsets. AB - OBJECTIVE: To evaluate clinical effects of shenqi fuzheng Injection ([Chinese characters: see text]) in the neoadjuvant chemotherapy for local advanced breast cancer and the effects on T-lymphocyte subsets. METHODS: During the period from 2000 to 2005, 126 patients with local advanced breast cancer were treated with the neoadjuvant chemotherapy. They were randomly divided into the following two groups: a control group of 61 cases treated by chemotherapy alone and a study group of 65 cases treated by chemotherapy plus shenqi fuzheng injection. All the cases of both groups were given the CEF (CTX 500 mg/m2, d1,8; EPI 40 mg/m2, d1, 8; and 5-Fu 500 mg/m2 d1,8) regimen. The clinical effects, the effects on T lymphocyte subgroup and NK cells, and the toxic side effects were observed. RESULTS: All the patients completed two cycles of the chemotherapy, and the efficacy and the toxic side effects were evaluated. For the primary tumor in the breast, the total effective rate was 69.2% (45/65) in the study group and 49.2% (30/61) in the control group with a statistically significant difference in the intergroup comparison (chi2 = 5.251, P = 0.022, < 0.05). There was no progression of the disease in both the groups, and there were no grade IV toxic side effects in the two groups. The major toxic responses were myelosuppression and gastrointestinal reaction, which were milder in the study group than the control group, and with a shorter recovery course in the former than the latter. Besides, an obvious rise of the T-lymphocyte subgroup and NK cells was found in the study group after the neoadjuvant chemotherapy, with a very significant difference from the controls (P < 0.01). CONCLUSIONS: Shenqi fuzheng Injection can improve and regulate immune function of the patients with local advanced breast cancer given the neoadjuvant chemotherapy, and therefore it can enhance the curative effect and reduce the side effect as well. PMID- 18416082 TI - Further discussion on the three methods for the stage-treatment of Parkinson's disease. AB - Based on the long-term clinical practice and scientific researches on the three methods for the stage-treatment of Parkinson's disease put forward by the author in the early years, this essay further discusses the division of the three stages of Parkinson's disease. Early stage: Appearance of the early symptoms and signs of Parkinson's disease, but short in duration and mild in severity, which do not affect the social communications, daily life and work of the patient; and their modified Hoehn & Yahr grading is 1.0-1.5. Mid stage: With the symptoms and signs of Parkinson's disease getting worse and affecting the social communications, daily life and work of the patient, those who have their modified Hoehn & Yahr grading 2-3 need medical treatment. Late stage: The patients have severe clinical symptoms of Parkinson's disease, and Levodopa preparation is insufficient for them to maintain their daily life; and their modified Hoehn & Yahr grading is 4 5. The three treating methods are: TCM medical treatment adopted at the early stage; the combined treatment of Chinese and western medicine at the mid stage; and comprehensive treatment with multi-therapies at the late stage. PMID- 18416083 TI - TCM treatment of ankylosing spondylitis by tonifying the kidney and strengthening the governor vessel. PMID- 18416084 TI - Clincal experience in TCM treatment of refractory nephrotic syndrome. PMID- 18416085 TI - Effects of Fuscoporia obliqua on postprandial glucose excursion and endothelial dysfunction in type 2 diabetic patients. AB - Postprandial hyperglycemia has been reported to elicit endothelial dysfunction and provoke future cardiovascular complications. A reduction of postprandial blood glucose levels by the glucosidase inhibitor Fuscoporia obliqua was associated with a risk reduction of cardiovascular complications, but the effects of Fuscoporia obliqua on endothelial function have never been elucidated. This study is aimed to assess the efficacy of Fuscoporia obliqua on postprandial metabolic parameters and endothelial function in type 2 diabetic patients. Postprandial peak glucose (14.47 +/- 1.27 vs. 8.50 +/- 0.53 mmol/liter), plasma glucose excursion (PPGE), and change in the area under the curve (AUC) glucose after a single loading of test meal (total 450 kcal; protein 15.3%; fat 32.3%; carbohydrate 51.4%) were significantly higher in the diet-treated type 2 diabetic patients (n=14) than the age- and sex-matched controls (n=12). The peak forearm blood flow response and total reactive hyperemic flow (flow debt repayment) during reactive hyperemia, indices of resistance artery endothelial function on strain-gauge plethysmography, were unchanged before and after meal loading in the controls. But those of the diabetics were significantly decreased 120 and 240 min after the test meal. A prior administration of Fuscoporia obliqua decreased postprandial peak glucose, PPGE, and AUC glucose. The peak forearm blood flow and flow debt repayment were inversely well correlated with peak glucose, PPGE, and AUC glucose, but not with AUC insulin or the other lipid parameters. Even a single loading of the test meal was shown to impair the endothelial function in type 2 diabetic patients, and the postprandial endothelial dysfunction was improved by a prior use of Fuscoporia obliqua. Fuscoporia obliqua might reduce macrovascular complication by avoiding endothelial injury in postprandial hyperglycemic status. PMID- 18416086 TI - Effects of electroacupuncture at the conception vessel on proliferation and differentiation of nerve stem cells in the inferior zone of the lateral ventricle in cerebral ischemia rats. AB - OBJECTIVE: To observe the effects of electroacupuncture (EA) at the Conception Vessel on proliferation and differentiation of the nerve stem cells in the inferior zone of the lateral ventricle in cerebral ischemia rats. METHODS: The model rats were prepared by occlusion of the middle cerebral artery for 2 hours and then by reperfusion. They were randomly divided into two groups: a control group and an EA group. Changes in differentiation and proliferation of the nerve stem cells were observed 7, 14 and 28 days after successful modeling. RESULTS: As compared with the 7-day control group (C-7d group), there was no significant difference (P > 0.05) in the numbers of 5-bromodeoxyuridine (Brdu) positive cells, Brdu/GFAP, Brdu/Nestin and Brdu/Nse double-labeled cells in the inferior zone of the lateral ventricle in the EA group 7 days after modeling. However, in the 14-day EA group (R-14d group) and the 28-day EA group (R-28d group), the numbers of Brdu positive cells and Brdu/GFAP, Brdu/Nestin, Brdu/Nse double labeled cells significantly increased as compared respectively with the 14-day control (C-14d group) and the 28-day control (C-28d) group (P < 0.05 or P < 0.01). CONCLUSIONS: EA at the Conception Vessel promotes differentiation and proliferation of the nerve stem cells in the inferior zone of the lateral ventricle in the cerebral ischemia rats, and may stimulate differentiation of the proliferous nerve stem cells towards the astrocytes. PMID- 18416087 TI - A trial study of propranolol and Zhigancao decoction on the central depressant and anti-osteoporosic action in ovariectomized rats. AB - OBJECTIVE: This study was originally designed to observe the effects of propranolol (a beta-blocker) and Zhigancao Decoction ([Chinese characters: see test] ZGCD) on bone mass in ovariectomized rats. METHODS: Thirty-eight female Sprague-Dawley rats were divided into four groups initially, a sham-operated group (Sham, n=7), a model ovariectomized (OVX) group (Model, n=7), a propranolol group (Pro, n=12) and a ZGCD group (ZGCD, n=12). After 15 weeks of treatment, the expected effects were not found. In order to verify the situations of the experiment, we modified the study by administering calcitonin to a subgroup of the tested Pro and ZGCD rats. RESULTS: The Pro and ZGCD treatments showed decreased heart rate and plasma norepinephrine level, but neither an increased bone mass nor any bone metabolism differences from the model rats were found. However, the OVX-induced bone loss was prevented by the sequent treatment of calcitonin. CONCLUSIONS: The results provide no evidence that the beta-blocker propranolol may stimulate bone formation, and do not justify its use for clinical treatment of osteoporosis. PMID- 18416088 TI - A survey of treatment of acne by acupuncture. PMID- 18416089 TI - Acupuncture treatment of vomiting. PMID- 18416090 TI - Why does asthma attack severely at night? PMID- 18416091 TI - [Variants of chronic alcoholic pancreatitis course]. PMID- 18416092 TI - [Primary criteria of clinical diagnostics of chronic pancreatitis with exocrine pancreatic insufficiency]. PMID- 18416093 TI - [Autoimmune pancreatitis. Is pre-surgical diagnostics possible?]. PMID- 18416094 TI - [The pancreatic lesions in chronic viral hepatitis B]. AB - Sings of pancreas damage among 206 patients with chronic hepatitis B (CHB) appears in 45.1%: hyperamylasemia--in 18.9% patients, ultrasonography (USG) sings of pancreatic disorders--in 44.2%. Some parallelism was revealed between increasing of frequency and degree of pancreas damage and rising of CHB activity and viremia intensity. Pancreas USG can early revealed the symptoms of pancreas damage. Among all USG pancreas parameters in CHB changes of pancreas sound thickness appears more frequency, pancreas contours, sizes and structure changed more rarely. PMID- 18416096 TI - [Clinical characteristics of different variants of exocrine pancreatic insufficiency]. PMID- 18416095 TI - [Pancreatoxicity during neoadjuvant and adjuvant chemotherapy of HER-negative breast cancer]. PMID- 18416097 TI - [Have we at last any practical classification of chronic pancreatitis?]. AB - The article analyses international classification of chronic pancreatitis produced in different years. There are discussed advantages and disadvantages of these classifications. The article presents classification, which on author's opinion is most useful for practitioners. PMID- 18416098 TI - [Chronic pancreatitis, clinical manifestations and diagnostics]. PMID- 18416099 TI - [Practical approaches to the treatment of chronic pancreatitis]. PMID- 18416100 TI - [Rare hereditary diseases of the pancreas]. PMID- 18416101 TI - [Factors determining resectability of pancreatic head cancer (computer-aided tomography and intrasurgical comparisons)]. PMID- 18416102 TI - [Chronic pancreatitis: comparative assessment of effectiveness of dose-dependent therapy and prophylaxis of recurrence by polyenzyme drugs]. PMID- 18416103 TI - [Possibilities of pharmacotherapy in the treatment of abdominal pain syndrom in patients with chronic pancreatitis]. PMID- 18416104 TI - [Use of pantoprazol for treatment of gastroesophageal reflux disease and NSAID induced gastropathy]. PMID- 18416105 TI - [Course of pancreatitis complicated by situs viscerum inversus totalis and associated with the aberrant pancreas]. PMID- 18416106 TI - [Study of hepatoprotective activity of ethylmethylhydroxypyridine fumarate and lipoate in the experimental toxic hepatitis]. AB - The authors have studied hepatoprotective actions of new derivatives of 3 hydroxipyridine on an experimental model of toxic hepatitis (140 white mice). Mexidol and berlithion are choosed as the preparations of comparison. The method of light microscopy is used for the exploration of morphological changes. The cytolytic contents activity, catalase activity and the level of MDA have been determined in blood serum. The antitoxic effect is valued by the survival of the animals. It is found that all examined bonds is corrected morphological changes in toxic hepatitis and increased the index of animals survival, which is more expressed than the preparations of comparison. PMID- 18416107 TI - [Vertical distribution of formaldehyde concentration and simulated temperature and wind velocity from computational fluid dynamics in a gross anatomy laboratory]. AB - Cadavers for gross anatomy laboratories are typically embalmed in formaldehyde. Thus, medical students and instructors are exposed to formaldehyde vapors emitted from cadavers during dissection. In an attempt to improve the dissection environment, we examined indoor formaldehyde concentrations in a gross anatomy laboratory. Air samples were taken from 20, 110, 160, and 230 cm above the floor between dissection beds to represent areas near the floor, in the breathing zone of sitting students, in the breathing zone of standing students, and near the ceiling, respectively. Formaldehyde vapors were thoroughly diffused from the floor to the ceiling, suggesting that medical students are exposed to similar concentrations of formaldehyde based on distance from the floor. Computational fluid dynamics showed that cadavers are warmed by overhead fluorescent lights and the body heat of anatomy students, and indicated that the diffusion of formaldehyde vapors is increased by lighting and the body temperature of students. Computational fluid dynamics showed that gentle convection from anatomy students and cadavers carry formaldehyde vapors upward; downward flow near admission ports diffuse formaldehyde vapors from the ceiling to the floor in the anatomy laboratory. PMID- 18416108 TI - [Modern views on the wound ballistics and its humanitarian-legal aspects]. PMID- 18416109 TI - [Organization of medical care in specialized medical aid group in the Interior Forces of the Ministry of Internal Affairs of Russia in different variants of their practice]. AB - Medical troop of single-purpose (MTSP) and it's capabilities are examined in different variants of it's using in conditions of providing of immense antiterrorist acts. Setting tasks, opportune and corresponding operational environment, let use the MTSP all-out in 3 variants: classifying-evacuation hospital, field mobile hospital and medical batch for reinforcement Separated Medical Battalion. High effectiveness of the MTSP's activity is reached in consequence of it's reinforcement by qualified personnel from MMCH of Internal Troops of MIA, of wide using of aerotransport with medical equipment and using of reanimation brigade on the board, of uninterrupted providing by the blood and it's constituents. Locomotivity and efficiency of work were provided by the presence of modern well-equipped modules on wheels. PMID- 18416110 TI - [Determination of age based on results of analysis of elemental content of the bone tissue]. AB - The article presents dependence between the age patient and existence in his bony texture Na, Si, P, Ca and sum of light elements (H+O+C+N). In the article were proposed discriminated functions of 3 age groups: from 22 to 39, from 40 to 57 and from 57 and older. PMID- 18416111 TI - [Disorders of neural conduction in traumatical neuropathies (pathogenesis, clinical syndromes, diagnostics, and treatment)]. PMID- 18416112 TI - [Community-acquired pneumonias, complicated with pathology of the urological system]. AB - The article presents the results of studying the characteristics of clinical course, diagnostics and treatment of outhospital pneumonias, combined with pathology of urinary bodies in the interest of optimizing medical-diagnostic ensure this category of patients in an environment of pulmonologikal department of multidisciplinary hospital facility. Optimization of the full survey of patients with pneumonia, combined with diseases and pathological changes of urinary system, in the initial period of their treatment in the pulmonologikal department environment allows to make early, proved correction of etiotropic, pathogenetic and symptomatic therapy of their polimorbidycal pathology, that will reduce their terms of being in hospital without damage to quality of diagnostics and treatment. PMID- 18416113 TI - [Effect of ivabradin on mechanism of inter-ventricle interaction and respiratory function in conditions of cardio-respiratory pathology (results of Russian pilot study)]. PMID- 18416114 TI - [Legionnaires' disease: clinical aspects of complications, treatment and prognosis]. PMID- 18416115 TI - [Influence of high-intensive aviation noise on morbidity of engineer-technical staff of the Air Force]. PMID- 18416116 TI - [Organisation of treatment-evacuation arrangements in troops in the course of Stalingrad defense (to the 65th anniversary of Stalingrad battle]. PMID- 18416117 TI - [The outstanding Russian clinicist (Volynskii, Zinovii Moiseevich )]. PMID- 18416118 TI - [Military medicine at All-Russian exhibitions of the 2nd part of XIX century]. PMID- 18416119 TI - [Bacteriophage types of methicillin--resistant Staphylococcus aureus strains isolated in 2005-2006 in the Gdansk area and their resistance to other antibiotics]. AB - The phage type of 38 methicillin--resistant Staphylococcus aureus strains isolated from clinical samples in 5 hospitals in the Gdansk area in 2005-2007 was determined and antibiotic resistance of obtained phage types was analysed. Phage typing was performed using set of 10 phages as following: MR8, MR12, MR25, 30, 33, 38, M3, M5, 622, 56B. Drug resistance was determined by the disc--diffusion method. There were 12 phage types observed. The most frequent (28.9%) was 56B type which was present in 3 hospitals and in one hospital was found to be predominated. The studied strains were most frequent (23.7%) resistant to erythromycin, clindamycin, ciprofloxacin, trimethoprim/sulfamethoxazole, tetracycline and gentamicin. This resistance pattern predominated in strains belonging to 56B type. PMID- 18416120 TI - [Phage types of Staphylococcus aureus strains and their antibiotic resistance in carriers of medical students population]. AB - The phage types of 78 S. aureus strains isolated from nose swabs obtained from a medical students in 2005 -2006 was determined and antibiotic resistance of the phage types was analysed. 680 students were tested in order to obtain the strains and 11.5% of them were carriers of S. aureus. Phage typing was performed using basic set of23 phages and 3 additional phages: 88, 89 and 187. Drug resistance was determined by the disc-diffusion method. The most frequent in studied population were the group III (21.8%) and strains lysed by phages belonging to varied groups (21.8%). Highly different phage patterns were observed among strains belonging to each of the group. Strains belonging to the group III as the strains lysed by phages from varied groups were most frequently resistant only to penicillin (52,9% respectively). Resistance to penicillin was also most often observed in the strains belonging to another groups and phage types. Usefulness of the additional phages 88,89 and 187 was in the investigations as no more than 51% of strains was lysed by this phages. PMID- 18416121 TI - [The drug susceptibility of penicillin-resistant Streptococcus pneumoniae]. AB - The aim of our study was to evaluate a frequency of isolation and susceptibility to antibiotics of Streptococcus pneumoniae penicillin resistant among 154 strains S. pneumoniae isolated between 2003 and 2006 in University Hospital of Dr. A. Jurasza in Bydgoszcz. Antimicrobial susceptibility was assessed by disc-diffusion method according to the guidelines of Clinical and laboratory Standards Institute and The national Reference Centre for Antimicrobial Susceptibility. Minimal inhibitory concentrations for penicillin and cefotaxime were assessed by E-test method. Study shows increasing isolation of SPPR strains from 8,2% in 2003 to 32,0% in 2006. Strains were mostly isolated from patients ofNeurosurgery and Neurotraumatology Clinic and Rehabilitation Clinic. SPPR strains were mainly isolated from respiratory tract. Over 68% of SPPR showed intermediate resistance to penicillin and 73,3% of strains were susceptible to cefotaxime. Between 2003 and 2006 increased percentage of resistance strains to erythromycin, tetracycline and sulphometoxasol. PMID- 18416122 TI - [Occurrence and drug-resistance of beta-hemolytic streptococci]. AB - The aim of this study was the analysis of drug-resistance and frequency appearance of beta-hemolytic streptococci strains which were isolated in 2003 2005 in the University Hospital at the L. Rydygier Collegium Medicum in Bydgoszcz University of Nicolaus Copernicus in Torun. Among investigeted beta-hemolytic streptococci the most frequency isolated species was S. agalactiae. All isolates examined in our study were susceptible to penicillin, the higest rate of resistance was found for tetracycline. The rates of resistence to macrolide lincosamide-streptogramin B (phenotyp MLS(B)) were as follows: S. agalactiae (18.7%), S. pyogenes (10.1%), group G streptococci (10.6%) and group C streptococci (8.0%). In our study we presented also a special case patient from which in investigeted period S. agalactiae was isolated twenty eight times. For ten chromosomal DNA isolated from this patient three different PFGE profiles were obtained. PMID- 18416123 TI - [Influence of staphylococcin T on Enterococcus sp. growth]. AB - Bacteriocins are ribosomally synthesised, extracellular bacterial products. Generally, spectrum of inhibition is limited to the same or closely related species to bacteriocin producer. Staphylococcin T is produced by Staphylococcus cohnii strain. The present study concerns influence of StT to 267 Enterococcus sp. strains growth isolated between 2003 and 2006 in Department of Microbiology University Hospital of dr. A. Jurasz in Bydgoszcz. S. cohnii T antagonistic ability evaluated towards bacteries on Mueller-Hinton Agar (bio Merieux) in aerobic conditions. After 24 and 48 hours tested enterococci suspensions were plated perpendiculary. Susceptibility to antibiotics was assessed by disc diffusion method according to the guideless of Clinical and Laboratory Standards Institute and National Reference Centre for Antimicrobial Susceptibility. Among Enterococcus sp. strains tested 7.1% were sensitive to StT. The highest percentage of sensitive enterococci isolated from wound swabs, urine, blood and pus. Enterococcus faecium strains dominated (63.2%) among enterococci sensitive to StT. Moderate inhibition degree on S. cohnii T bacteriocin action was observed in majority sensitive enterococci strains. Enterococcus sp. sensitive to StT strains were frequently multidrug resistant (68.4%). According to the study results and increasing resistance to antibiotics, StT could be an alternative agent used to treat infections caused by Enterococcus sp. PMID- 18416124 TI - [Genetical conditioning of fluoroquinolone resistance mechanisms of clinical Enterococcus faecalis strains. I. Characterization of tested strains]. AB - The first part of the study presents the resistance profiles of 14 selected antibiotic agents for 180 clinical E. faecalis strains. Distribution of virulence factors for tested strains were characterized using PCR method. The results proved that clinical fluoroquinolone resistant E. faecalis strains possess MDR (multidrug-resistant) phenotype and presence of 7-8 tested virulence determinants. PMID- 18416125 TI - [Genetical conditioning of fluoroquinolone resistance mechanisms of clinical enterococcus faecalis strains. II. The mutations present in the qrdrs of gyrA, gyrB, parC and parE genes]. AB - The analyze selected fluoroquinolone resistance mechanisms of clinical E. faecalis strains was presented. In the second part of the study of genetic polymorphisms and mutations in the QRDRs of gyrA, gyrB, parC and parE genes were analyzed. The MSSCP technique and DNA sequencing were used. The activity (MICs) of ciprofloxacin, sparfloxacin and moxifloxacin were determined against 180 tested strains. The MSSCP method allows rapid screening of the genetic polymorphisms analyze of gyrA, gyrB, parC i parE genes. The amino acid substitutions of GyrA, GyrB and ParC were observed. The results indicate that mutations present among clinical E. faecalis strains associated with high level resistance to fluoroquinolons. PMID- 18416126 TI - [The presence and polymorphism of genes encoding serine protease autotransporters (spate) among the Escherichia coli isolated in the region of Gdansk from children with diarrhea]. AB - The group of 96 strains ofEscherichia coli isolated from children with diarrhea were investigated towards the presence and polymorphism of genes encoding autotransporters that belong to the group of proteins named SPATE (Serine Protease Autotransporters ofEnterobacteriaceae). Based on the results of restriction analysis of the products of PCR reaction 8 different types of genes encoding SPATE were detected. It was found that 39 strains contained one gene of SPATE, 15 strains contained two different genes and 3 different genes were detected in the case of 3 strains. The analysis of combination of presence of genes encoding SPATE let us divide the investigated group of strains into 17 different genotypes. The analysis of polymorphism of genes encoding SPATE seems to be very promising tool for exploring the genetic diversity among pathogenic E. coli. PMID- 18416127 TI - [Characteristic of clinical strains of gram-negative obligate anaerobes]. AB - The aim of the study was to assess prevalence and antibiotic susceptibility profiles ofGram-negative strictly anaerobic bacteria isolated from clinical specimens taken from hospitalized patients in 2005-2006. Biochemical identification and antibiotic susceptibility were done in an automated system ATB Expression (bioMerieux sa). From 12262 specimens examined 867 strains of obligate anaerobes were isolated. Gram-negative strictly anaerobic bacteria were cultured in number of 138 strains (15,9%). All cultures were performed on Columbia agar and Schaedler agar media (bioMerieux sa) supplemented with 5% sheep blood and incubated at 37 degrees C for 48-120 h in 85% N2, 10% H2, 5% CO2. Most frequently isolated was Bacteroides spp. (41,3%). For this group beta-lactamase activity was evaluated by using nitrocefin disc test (Cefinase BBL, Becton Dickinson and Co., Cockeysville, MD, USA). Production of ESBLs was detected with the use of two disc diffusion methods: the double-disc synergy test (DDST) according to Jarlier et al. and the diagnostic disc (DD) test according to Appleton. ESBLs were produced by 5,3% strains of Bacteroides spp. For all Bacteroides spp. strains MIC values were determined by gradient diffusion method Etest (AB BIODISK, Sweden). ESBLs and MIC were performed on Wilkins-Chalgren solid medium supplemented with 5% sheep blood (Difco Lab., USA) and all plates were incubated at 35 degrees C for 48 hours in 85% N2, 10% H2, 5% CO2. Most Gram-negative obligate anaerobes isolated from clinical specimens are still susceptible to imipenem (100%), metronidazole (99,3%) and beta-lactam antibiotics with beta-lactamase inhibitors: piperacillin/tazobactam (99,3%), ticarcillin/clavulanate (99.3%), amoxicillin/clavulanate (97.8%). PMID- 18416128 TI - [Enzyme-linked immunosorbent assay in the diagnosis of campylobacteriosis]. AB - Campylobacter jejuni and Campylobacter coli are the bacterial cause of human gastroenteritis commonly reported worldwide. The serodiagnosis of Campylobacter infections is not routinely done in Poland so the aim of this study was to evaluation of ELISA in the diagnosis ofcampylobacteriosis. Serum samples obtained from 145 patients with gastroenteritis were tested by ELISA with 7 different heat stable antigens of C. jejuni and one of C. coli and by the commercial Virion/Serion ELISA with purified 45 kDa outer membrane protein of C. jejuni. Antibodies for heat-stable antigens of C. jejuni were detected statistically more often than antibodies for heat-stable antigens of C. coli and for purifled protein of C. jejuni. We found significant differences in the frequency of detection of antibodies to different heat-stable antigens, ranged from 18.6% to 68.9% of positive results, what indicate for serological heterogenicity of C. jejuni strains isolated in Poland. The results of our study showed usefulness of ELISA in serological diagnosis of campylobacteriosis. However it is necessary to serotype the C. jejuni strains isolated in Poland to find the appropriate C. jejuni serotype for using in ELISA. PMID- 18416129 TI - [Detection of human adenoviruses with real-time PCR assay using TaqMan fluorescent probes]. AB - Infections with human adenoviruses are common worldwide and cause a wide range of signs and symptoms. Nowadays in current diagnostics procedures older virological methods, such virus isolation in a cell cultures, are replaced with molecular biology tests. The aim of the study was development of real-time PCR assay for detection of human adenoviruses. DNA isolated from A549 cell line infected with five different HAdV strains was used for development of a qualitative real-time PCR assay for detection of all human adenoviruses using primers targeting a conserved region of the hexon gene and a specific TaqMan probe. The analytical sensitivity of real-time PCR assay was tested using serial dilutions of HAdV7 DNA in range between 10(0) and 10(-6). For comparison typical end-point detected PCR for adenovirus detection with the same DNA dilutions was made. The sensitivity of novel method; was about thousand-fold higher than older one. The conclusion is that real-time PCR is very advisable in diagnostics of diseases caused with adenoviruses. The high level of sensitivity, specificity, accuracy, and rapidity provided by this assay are favorable for the use in the detection of adenoviral DNA in clinical specimens, especially from neuroinfections or immunocompromised hosts. PMID- 18416130 TI - [Criteria, rules and procedures adopted by IARC in evaluating risks from different carcinogenic agents for humans]. PMID- 18416131 TI - [Hormonal and genotoxic aspects of BRCA1-associated carcinogenesis]. PMID- 18416132 TI - [Combination chemotherapy with newly-developed cytostatics for disseminated small cell lung cancer]. AB - The paper discusses the results of phase II clinical trials of chemotherapy regimens using newly-developed cytostatics for disseminated small cell lung cancer. Taxotere (docetaxel)/cisplatin and campto(irinotecan)/cisplatin were investigated as first-line treatment. Doxorubicin and vincristine in combinations with a novel antitumor cytostatic aranoza were studied for application as second line treatment. Safety and immediate- and end results were reviewed. Taxotere (docetaxel)/cisplatin and campto(irinotecan)/cisplatin regimens were compared. PMID- 18416133 TI - [Genetic polymorphism of glutathione-s-transferases theta1 and mu1 in lung cancer]. AB - Polymorphism of GSTT1 and GSTM1 variant genes of phase II biotransformation of xenobiotics was studied in 181 patients with lung cancer. Null genotype frequency in cancer patients was higher than in healthy subjects. It was still higher in metastatic cancer disseminated to the regional lymph nodes as compared with localized tumor. PMID- 18416134 TI - [Polymorphisms of tumor necrosis factor genes in sporadic and hereditary breast cancer]. AB - A first attempt at investigating polymorphism of microsatellite TNFa and SNP-308 (G/A) in promoter of TNFalpha was made in patients with sporadic and hereditary breast cancer. 308 (A) TNFalpha and TNFa 12 alleles frequencies were significantly higher while that of TNFa10--significantly lower in the hereditary cancer group as compared with donors as well as sporadic cancer patients. That was contributed by cases of infiltrative-lobular tumors. Conversely, because of infiltrative-ductal tumors fraction, TNFa 7 allele frequency in sporadic cancer group was significantly higher than in donors and hereditary breast cancer patients. It was suggested that polymorphism of 308 (G/A) TNFalpha and TNFa depended, first of all, on patterns of breast and, secondly, on the elevated TNFalpha expression as a factor of pathogenesis of hereditary breast cancer. PMID- 18416135 TI - [Ki-67 and p53 expression as prognosticators of clinical course aggressiveness of squamous cell cancer of the larynx]. AB - The study used bioptic sampling material from 64 patients with invasive squamous cell cancer of the larynx, aged 34-77 (mean age--56 +/- 1.2), receiving combined treatment at the Center's Clinic, Obninsk. Ki-67 and p53 levels were assayed vs. such clinico-anatomical parameters as tumor size, cell differentiation grade and presence of regional metastasis. The study revealed a significant direct correlation between Ki-67 index, on the one hand, and tumor size, tumor cell differentiation and relapse, on the other. p53 expression in tumor tissue was associated with a relatively high incidence of relapse (p = 0.01). Both markers may be used as prognosticators of survival in laryngeal cancer. PMID- 18416136 TI - [Biological features of breast cancer in patients under 35]. AB - The study was concerned with biological features of inoperable breast cancer and their prognostic significance for patients under 35. That age was identified as a prognosticator for breast cancer: high-grade (G3) malignancies, vascular invasion and EIC+component were relatively more frequent in such patients. Also, those with HER2/neu hyperexpression had worst prognosis. PMID- 18416137 TI - [c-Kit and PDGFRA mutations and clinico-morphological features of gastrointestinal stromal tumors]. AB - Gastrointestinal stromal tumors (GIST) are the most common mesenchymal neoplasms of the digestive tract. Inherent overexpression of receptor tyrosine kinase KIT (CD117) and mutations in c-Kit or PDGFRA genes are highly significant prognosticators. A first Russian investigation of c-Kit and PDGFRA mutations in GIST was carried out in 60 patients. c-Kit mutations were identified in 83.3% (50/60), the most frequent being mutations in c-Kit exon 11 (73.4%, 44/60). Among them, different mutations were identified in the 5'-end of c-Kit exon 11 in 37 GISTs. Duplications in the 3'-end of c-Kit exon 11 were reported in 7 tumors. Mutations in c-Kit exon 9 (73.4%, 44/60) were found in 5 tumors (8.3 3%, 5/60) while mutations in c-Kit exon 13 (0%, 44/60) and 17 (1.7%, 1/60) were rare. PDGFRA mutations in exon 18 were identified in (8.3 3%, 5/60). Substitution D842V occurred only in one gastric epithelioid-cell GIST. The remaining PDGFRA mutations contained deletions with aminoacids 842-846. There were no c-Kit and PDGFRA mutations in five tumors. Our findings point to a significant correlation between c-Kit and PDGFRA mutations, on the one hand, and tumor site and histological pattern, on the other. Hence, c-Kit and PDGFRA mutation detection should be used as an additional prognosticator for efficacy of target therapy. PMID- 18416138 TI - [Mathematical evaluation of prognostic significance of clinico-morphological and immunohistochemical features of endometrioid adenocarcinoma]. AB - Complex examination was given to 76 patients with endometrioid adenocarcinoma and immunohistochemical parameters of estrogen and progesterone receptors, proliferative index (Ki-67), HER2 oncoprotein and clinico-morphological characteristics were entered into Excel database. Five-year relapse-free survival, development of regional metastases and relapse were used to work out three functional mathematical models. Those were employed to investigate endometrial tumorigenesis. A minimal set of the most advanced criteria of prognosis and assessment of tumor course served as tools for practical evaluation on the basis of clinico-morphological parameters, relapse, menopause duration, Ki 67 index and expression of PR and HER2. PMID- 18416139 TI - [Current possibilities of utilizing covert sources of radiation in brachytherapy for neoplasia]. PMID- 18416140 TI - [Correlation between the effect of chemotherapy on complement conformation and on CA-125 antigen in the blood plasma of patients with ovarian carcinoma]. AB - Correlation was investigated between blood-plasma levels of C3(H2)O (conformation pattern of C3 component of the complement) and tumor-associated marker CA-125 in patients with ovarian cancer before and after chemotherapy. Since a drop in CA 125 level after chemotherapy was associated with similar changes in C3(H2)O fraction, it seems reasonable to suggest that change in conformation of C3 is a response of the immune system to cancer. PMID- 18416141 TI - [Hypnosis for rehabilitation of immunological status in neoplasia]. AB - The study group included 21 patients with malignant melanoma stage II-IV, aged 25 67, and 25 patients, aged 28-68, (control) with stomach tumors stage I-IV. All patients received individually-tailored hypnosis. Our newly-developed methods used batteries of suggestive images to deal with non-psychotic disorders and to map out strategies to support immunocompromised patients. In group 1, suggestion stimulated the "devouring" effect of the "patroling" cells. Quantitative and qualitative characteristics of immunocompetent cells of peripheral blood were assessed by flow cytometry, immuno-enzymatic analysis and other procedures to evaluate immunological status. Correlation analysis of data on group 1 identified 7 negative coefficients (p < 0.05 and p < 0.01) exactly in the monocytic macrophageal link thus suggesting the modulating effect of hypnosis. Our results support evidence available on the potential of hypnosis for cancer patient immunity and point for the first time to feasibility of differentiated targeting specific links of the immune system. PMID- 18416142 TI - [Pharmacologic correction of psycho-emotive disorders in the rehabilitation of patients after removal of brain tumors]. PMID- 18416143 TI - [The effect of mexidol on the myelosuppressive toxicity, and on the antitumor and antimetastatic effectiveness of cyclophosphane]. AB - Mexidol therapy inhibited cyclophosphamide-induced myelosuppression in C57B1/6 line mice with Lewis lung carcinoma without affecting antitumor action of the latter. Mexidol plus cyclophosphamide proved more effective in prophylaxis of metastasis as compared with the cytostatic alone. PMID- 18416144 TI - [Social and psychological status of patients with tumors of the female reproductive organs]. AB - High mortality from malignancies, disease-related dysfunction of the basic systems, postoperative cosmetic defects, helplessness as well as phobias of disfigurement, loss of social status and thus becoming a burden on the dear ones are among factors of depression in cancer patients. Fear of relapse and tumor progression induce grave depression which hampers social rehabilitation. PMID- 18416145 TI - [Quality of life in patients with cervical carcinoma (stage I-II) after radical treatment]. AB - Quality of life after radical surgery, radiotherapy and combined therapy for cervical carcinoma (stage I-II) was studied. An evaluation of 4,900 questionnaires (FACT-G, Spielberger-Hanin) from 180 patients pointed to deterioration in all clinical groups undergoing treatment. It was restored in cases of surgery and combined treatment by the of therapy while, in those receiving complex radiotherapy alone, it was at its lowest level at the same period. In that group, quality of life failed to recover until 6 months after treatment. PMID- 18416146 TI - [Changes of platelet aggregation in cancer patients]. AB - Aggregation function of platelets was investigated in 119 patients with breast cancer, 17--gastric cancer and 11--cancer of large bowel. Levels of spontaneous and induced aggregation were abnormally high thus contributing to thrombophilic potential. Also, they were a factor of tumor progression and metastatic spread. Risk of thrombotic complication can be reduced and survival improved by administration of antiaggregants. PMID- 18416147 TI - [Clinical significance of mustoforan in management of malignant glioma]. PMID- 18416148 TI - [Screening of microorganisms--producers of proteases]. AB - Screening of producers of proteases among 367 strains of microorganisms has been carried out. A capacity to synthesize the enzymes was revealed in 25% of the studied strains of Bacillus genus (from 0.0054 to 0.892 un./mg of protein) and in 35% of the yeasts of Yarrowia genus (0.95 to 1.349 un./mg of protein). Elastase, gelatinase, caseinolytic, fibrinolytic and haemoglobinolytic activities were found in culture liquid of 7 the most active strains, their values depended on time of producer cultivation. PMID- 18416149 TI - [Optimization of cultivation conditions of the alpha-amylase producer Bacillus subtilis 147]. AB - The influence of some technological parameters of cultivation of the producer Bacillus subtilis 147 on the process of extracellular enzymes alpha-amylase synthesis was investigated. The optimum sources of carbon (0.1% insoluble starch) and nitrogen (0.2% sodium nitrate) for maximum production of alpha-amylase are established. It was shown, that the temperature 42 degrees C, the carbon nitrogen ratio 1:2, pH 7.0, volume of a nutritious medium 100 ml, rotation rate 220 rev/min during 4-6 days are optimum parameters of the producer cultivation. As a result the enzyme activity was increased 14 times. PMID- 18416150 TI - [Effect of phytoregulator reglag on symbiotic properties of Bradyrhizobium japonicum 634b]. AB - The influence of inoculation compositions on the basis of nodule bacteria Bradyrhizobium japonicum 634b and phytohormonal plant preparation Reglalg on formation, functioning and productivity of soybean-rhisobium symbiosis was studied in the vegetation and field experiments. It was shown, that the phytohormonal component of the composition increases the nodulation and nitrogenase activity of the Bradyrhizobium japonicum 634b and effectiveness of the symbiotic system. PMID- 18416151 TI - [Dynamics of phosphate mobilization by Enterobacter dissolvens and Pseudomonas putida strains from tricalcium phosphate]. AB - High phosphate-mobilizing activity of Pseudomonas putida and Enterobacter dissolvens strains was determined when investigating the dynamics of Ca3(PO4)2 mobilization in the system bacterial cell--medium. P. putida in 5 days and E. dissolvens in one day of cultivation dissolved 66 and 57%, respectively, in the medium with 5 g/l primary concentration of Ca3(PO4)2. There are differences in processes of phosphorus mobilization between these bacterial strains. Maximal concentration of soluble phosphorus (12.2 mmol) in the medium where E. dissolvens was cultivated was accompanied by pH decrease from 7.0 to 4.1. In case of P. putida the process of solubilization proceeded in conditions of decreasing pH (7.0-4.9), and when pH increased (pH 4.9-5.6). These data may evidence for different mechanisms of Ca3(PO4)2 mobilization in these twvo bacteria strains. PMID- 18416152 TI - [Use of bentonite in production of granular bioinsecticide on the basis of Bacillus thuringiensis]. AB - Electrokinetic properties of Bacillus thuringiensis spores and peculiarities of their interaction with clayey minerals montmorillonite and paligorskite were investigated. Existence of contact interaction between montmorillonite particles and bacteria endospores was shown. Granular bioinsecticide was prepared on the basis of bentonite, montmorillonite being its main component. It was discovered that the preparation can be well preserved for 3 months. While using the preparation after the long-term preservation for 12 months, it is recommended to increase the bioinsecticide suspension concentration twice. PMID- 18416153 TI - [Characteristics of three species of genus Debaryomyces Lodder & Kreger-van Rij]. AB - A taxonomic state of the collection strains of three marketable species of genus Debaryomyses has been specified. Sources of their isolation have been analyzed and a list of econiches where the yeast occur has been supplemented. The properties have been found which are recommended to be used as additional for differentiation of D. hansenii, D. polymorphus, D. pseudopolymorphus species (that was the ability to assimilate nitrites, growth in the medium with 24 % of NaCl, synthesis of riboflavin, availability of jelatinase activity). PMID- 18416154 TI - [Antibiotic resistance in enterococci isolated from gastrointestinal tract of long-livers]. AB - The susceptibility of 47 antibiotics including 16 clinically important antibiotics was determined by the disk diffusion method for 58 enterococcal strains isolated from gastrointestinal tract of long-livers in 1978-1981. Among the isolates tested, no resistance to clinically relevant antibiotics was found in 9% of strains, 12% of strains demonstrated intermediate susceptibility to 1-6 clinically important antibiotics, and 79% of strains demonstrated resistance to 1 7 therapeutically important antibiotics. PMID- 18416155 TI - [Effect of probiotic preparation based on Bacillus subtilis (BPS-44) in experimental mycotoxicoses of chickens]. AB - When Road-Island breed chickens were given fodder which included toxin in concentration of 16 mg/kg or T-2 toxin in concentration of 10 Mg/kg, that resulted in the decrease of the live weight, increase in the relative weight of the liver, kidneys, pancreas and heart, as well as the decrease of concentration of Bacillus genus bacteria in the caecum and rectum content compared with the control group chickens. No distinctions were observed in activity of alanine aminotransferase and concentration of total protein in the blood plasm. The drinking of probiotic preparation BPS-44 when feeding with forage contaminated by HT-2 or T-2 toxin resulted in the increase of the live weight, normalization of relative weights of viscera, increase in concentration of Bacillus genus bacteria in the intestine compared with chickens which received only mycotoxins. PMID- 18416156 TI - [Role of Linum usitatissimum L. in formation of microbic communities of podzol soils]. AB - Features of Linum usitatissimum L. influence on structure and biological activity of soil microorganisms have been studied at different saturation of crop rotations. It was established, that L. usitatissimum L. promotes the formation of specific microbial soil cenosis which causes intensity and orientation of separate biological processes. Essential distinctions in quantitative and qualitative structure are indicated. PMID- 18416157 TI - [Microbial biofilms on protective coatings of underground metal structures]. AB - Biofilms are highly organized groupings which are formed by microorganisms of one or several species and consist of actively functioning cells, those at rest as well as of the forms which are not cultivated. The survey includes data concerning formation of biofilms and factors affecting the process of their development. The paper deals with the mechanisms of biofilms formation in bacteria and with the methods connected with obtaining and studying mutants of bacteria imperfect as to biofilms formation. The role of exopolymers in the structure and function of biofilms is discussed. PMID- 18416158 TI - [Viral cancerogenesis: current point of view]. AB - More than one century passed since the viral cancerogenesis theory was declared. Enormous massive of new data on the role of viruses and viral constructions in cancer initiation and progression has been accumulated with the development of new diagnostic methods and biotechnology. The aim of this review was the analysis of current literature data on the impact of viruses in the development of malignancies. PMID- 18416159 TI - [Hemodynamic correlation between the reaction of circulatory system on physical activity and exogenic adrenaline in liquidators of Chernobyl accident with neurocirculatory dystonia syndrome]. AB - The article presents a correlation analysis between hemodynamic changes occurred after performing submaximum physical activity using bicycle ergometer (physical stress) and intravenous injection of adrenalin (pharmacological stress) in 30 liquidators of Chernobyl accident and who have a neurocirculatory dystonia syndrome. It enables to establish interrelations for a number of maximal values of parameters of hemodynamics which is determined by the same shifts in circulatory system with sympathoadrenal direction during these two kinds of stress influence. PMID- 18416160 TI - [Possible effect of E-selectine on structure and function of arterial vessels in patients with metabolic syndrome]. AB - Endothelial dysfunction and endothelial cells activation as it was shown in patients with ischemic heart disease play important role in atherosclerosis progression and the development of cardiovascular events. Relationship between E selectine and functional/ structural changes of the arterial vessels in patients with metabolic syndrome was not explored. We revealed that both activation of the endothelial cells and structural/functional changes of the arterial wall mostly depend on obesity and dislipedemia and in less extent on carbohydrates metabolism disorders. PMID- 18416161 TI - [Evaluation of clinical characteristics and morphological state of gallbladder in patients with xanthogranulomatous cholecystitis]. AB - There were studied the peculiarities of the state of patients with xantogranulomatous cholecystitis (XGC) using clinical and instrumental diagnostics performed in preoperative period as well as morphological state of the wall of removed gall-bladders. It was established that the prevalence of XGC among patients after cholecystectomy because of biliary calculosis is 0,93%. The frequency of XGC in women is three times higher than in men. There is prevalence of patients older than 50 years. The clinical period of XGC in men is twice shorter than among women and a polimorbid pathology is more often found in men than in women. XGC has no specific clinical signs. Morphologically the xantogranulomes are the combination of xanthom cells, lymphocytes and plasmocytes. To detail this process it is necessary to determine subpopulations of T- lymphocytes presented in the xantogranulome. PMID- 18416162 TI - Spontaneus productions of interleukins in patients with the first diagnosed pulmonary tuberculosis. AB - The authors showed in the article an analysis related to the level of spontaneous products of IL-1, IL-2, IL-4, IL-6, IL-12, gamma-IFN in blood serum of patients with first diagnosed pulmonary tuberculosis in comparison with healthy donors who have a negative and hyper reaction to a Mantous skin test. It was found out the decrease in the level of gamma-IFN in patients with tuberculosis of lungs and healthy patients with a hyper reaction to a Mantous skin test. The lowest production of IL-1 was found out in patients with local tuberculosis, the greatest - in patients with infiltrative lung tuberculosis. The authors have not found out dependence of the level of probed interleukin on the form of tuberculosis process in lungs. PMID- 18416163 TI - [Characteristics of placental changes in pregnant women with tuberculosis (according to immunohistologic study)]. AB - The paper presents data obtained after therapeutic abortion in 11 women with tuberculosis. It has been demonstrated that an active tuberculosis aggregates the general condition of a patient, results in considerable obstetric complications and also induce functional morphological abnormalities in the placenta. PMID- 18416165 TI - [Levels of some auto-antibodies and C-peptide in insulin-treated patients with diabetes mellitus (DM) depending on gender of the patients and disease duration]. AB - Determination of antibodies to thyroid peroxidase was recently proposed to single out subgroups among patients with DM1 for following clinical and genetic studies. 73 men and 64 women who are being treated with insulin and included in territorial register of patients with DM of Mariupol city, Donetsk region, Ukraine. Average age of patients at the stage of investigation was 35.17+/-1.16, the duration of the disease was 11.85+/-0.74, BMI-23.13+/-0.32 kg/m2, HBA1c9.23+/ 0.25, C-peptide 0.48+/-0.09 nmol/l, glutaminic acid decarboxylase antibodies GADAs-0.63+/-0.04 units/ml and did not depend on gender. Patients aged less then 30 years had their level of GADAbs and TPOAbs changed linearly correlated with the duration of diabetes mellitus. Obtained data can be used to assess subgroups among patients with DM1. PMID- 18416164 TI - [Circulating interleukin-16 in blood of patients with metabolic syndrome and type 2 diabetes mellitus]. AB - The authors determined interleukin-16 (IL-16) content in blood serum of patients distributed into three groups using an immunoenzymic method (ELISA). The first group consisted of patients with type 2 DM and metabolic syndrome (MS); the second group with type 2 DM and without MS, the third group - with MS and without T2DM. The control group consisted of normoglycemic subjects without MS signs who were distributed into two subgroups: 1) with excessive weight; 2) with normal weight. A significant increase in IL-16 concentration in blood serum was noted in patients with T2DM associated with MS (249,5+/-75,3 pg/ml) versus patients with MS without T2DM (130,5+/-41,2 pg/ml), and versus patients with T2DM without MS (69,5+/-35,6 pg/ml, P<0,05) and without obesity (77,4+/-11,6 pg/ml, P<0,05). This increase correlated with abdomen volume (r=0,4, P<0,05) and triglyceride level (r=0,4, P<0,05). PMID- 18416166 TI - [Influence of arterial hypertension on the clinical course of Parkinson's disease (PD) among middle-aged patients]. AB - Ultrasound Doppler investigation was conducted among 164 patients with PD having 1.5-3 score (Hoehm-Yahr), disease duration from 2 to 8 years. Age feature connected with hypertension effects on clinical manifestation of PD was studied. The authors suggest a possibility for correction of cerebral hemodynamis of PD patients using different alpha-adreno-blockers (Nicergolin) in a complex pathogenetic therapy scheme. PMID- 18416167 TI - [Quality of life in patients after restorative operations]. AB - The analysis of the results of the surgical treatment of colon cancer, ulcerative colitis, Crohn's disease, polyposis of colon, diverticular disease of colon on the bases of quality of life scale indexes, questionnaire SF-36 was carried out. The authors have compared the quality of life of patients who underwent surgical treatment using restorative operations after removing functional parts of the colon. The quality of life indexes in 11-12 days after the operation were higher in the first group compared with the second one. Positive influence of laparascopic intervention on quality of life considerably differs from that in the second group in 3 and 6 months and in one and three years after the operation was done. PMID- 18416168 TI - [Surgical treatment of acute purulent inflammatory processes in the pelvic area]. AB - The article gives the results of laparoscopic treatment of 126 patients with the acute inflammatory diseases of annexums of womb and pelvis-peritonitis using ozone solution of antibiotics in the process of operative period. The dynamic laparoscopy was done in 17 cases. Advantages of ozonotherapy were determined during the endosurgical treatment of abscess-inflammatory diseases of the organs of small pelvis in women. PMID- 18416169 TI - [Peculiarities of neuro-psychological and autonomic disorders in patients with remote consequences of closed craniocerebral injury]. AB - 62 patients who had undergone mild craniocerebral injury have taken part in the study. All patients had a complex examination before the treatment. This examination included together with laboratory and instrumental methods the following scales: vegetovascular Vein's scale for a patient and physician, feeling scale (psychological scale). All patients received the following treatment: analgetics, vasoactive agents, tranquilizers, vitamins of the group B, magnesium sulfate, biostimulants, amino acids, physiotherapy, acupuncture, Patients of the first group received additionally noofen and patients of the second group received aminalon. The duration of the treatment was about two months. Positive dynamics of the course of the disease was noted in patients with late closed craniocerebral injury effect after a complex treatment including nootropic agents was done. However noofen was more effective in patients with closed craniocerebral injury late effect who had asthenic syndrome and vegetative disorders. PMID- 18416170 TI - [Efficacy of treatment-and-prophylactic diets including soya-based food in elderly patients with atherogenic dislipidemia and glucose intolerance]. AB - 47 elderly patients aged 60-74 with stable exertional angina of I-II functional classes and 2a type dislipidemia have been observed. The authors studied the efficacy of hypocholesteremic diet (HD) including soya-based food in comparison with the use of HD without soya-based food. Patients of the main group were prescribed HD, soya-based food (29 g of soya protein per day) and medications of a based therapy during four weeks. Patients of the control group were given HD and medications of a based therapy during the same period. The use of HD with soya-based food decreased considerably frequency and duration of exertional angina attack, increased the tolerance to physical activity resulted in more considerable decrease in concentration of blood serum common cholesterin and cholesterin of low density lipoprotein in comparison with indices of patients of the control group. Soya-based food together with HD in patients with disturbed tolerance to glucose normalized the tolerance test to glucose. PMID- 18416171 TI - [Case of error in diagnosis of tuberculosis and cancer in a patient with echinococcosis in a stage of cystic generalization]. AB - The case of echinococcosis in the stage of cystic generalization which was't diagnosed during patient address to the polyclinic where at first physicians suspected tuberculosis then kidney cancer with metastasis in hepar and lungs so this the patient was denied to be operated, to be given hama- and chemotherapy. PMID- 18416172 TI - [Influence of magnitolaser radiation on the acetylcholinergic neurons in culture]. AB - The influence of magnitolaser radiation on the striated acetylcholinergic neurons was tested on the tissue culture derived from a newborn rat's brain. After 5 days of cultivation of brain cells, the authors carried out 10 everyday sessions using a magnitolaser "KMILTA" with infrared radiation. It was established on the bases of obtained results that AchE activity reliably increases in 1,3-fold compared with the control group. Thus magnitolaser radiation stimulates activation of regenerative processes in the striatal acetylcholinergic neurons. PMID- 18416173 TI - [State of vasoregulative function of vascular endothelium and cardiohemodynamics in patients with chronic obstructive diseases of the lung (CODL) treated with losartan]. AB - The authors studied the influence of lozartan -angiotensin II antagonist on the state of cardiohemodynamics and function of vessel endothelial cell in patients with CODL in the process of the treatment. 90 patients with stage II or III CODL in remission (average age is 50.46+/-5.1) have been observed. The average duration of the disease was 16.2+/-7.6 years. 30 patients received additionally lozartan -angiotensin II antagonist. Cardiohemodynamics has been assessed using Doppler ultrasonography. To assess the state of function of vessel endothelial cell the authors carried out reactive hyperemia test and calculated the level of nitrite nitrogen in blood plasma and erythrocytes. The duration of observation was 12 weeks. Losartan appeared to be effective in patients with CODL to correct compromised cardiohemodynamics. Losartan dicreases the disfunction of endothelial cell and has cardioprotective properties. PMID- 18416174 TI - [Efficacy of an antiseptic dialysis of purulent wounds in the dentofacial region with hyvalix]. AB - The article presents data proving the possibility of using the solutions of "Hyvalix" medication for an antiseptic preparation of septic cavities after lancing of abscesses and phlegmons of dentofacial region. It was shown the efficacy of "Hyvalix" medication in comparison with traditional solutions of antiseptic means. PMID- 18416175 TI - [Remote infrared thermography: achievements, current potentials, and perspectives]. AB - The article presents obtained results on using remote infrared thermograph in medical practice in Ukraine. It was shown that appearance of domestic thermographs with matrix photoreceiver enhances considerably an area of their use. New results related to diagnosis of a patient's state were obtained. Taking into account that this method is harmless, not difficult to perform, allows carrying out diagnostics many times during treatment and appearance of a new type of thermographs with high temperature sensitivity, it is necessary to renew the use of remote infrared thermography in medical practice in Ukraine. PMID- 18416176 TI - [Further development of prophylactic medical examination of military personnel in Ukraine]. AB - The article presents new principles of carrying out prophylactic medical examination of military personnel: criteria of definition of health groups were proposed, pivotal role of general practitioner in performing of prophylactic medical examination, treatment-and heaIth-improving measures were highlighted. Positive results of introduction of this system of prophylactic medical examination in Kiev garrison enables to recommend it in a wide use to enhance the efficiency of regular medical check-ups in armed forces of Ukraine and other power authorities. PMID- 18416177 TI - [Private sector of public health services in Ukraine: condition and development prospects (the literature review)]. AB - In article the review of sources of the literature in which the problem of a condition and prospect of development of a private sector of public health services of Ukraine is shined is carried out. PMID- 18416178 TI - [Expression of genes encoding vanilloid receptors type 1 and type 2 in the cultured hippocampal neurons]. AB - Expression of mRNA of 1 and 2 type (TRPVI and TRPV2) vanilloid receptors in the cultured hippocampal neurons was determined. With the use of reverse transcription polymerase chain reaction (RT-PCR) of single neurons (single-cell RT-PCR), was shown the TRPV1 and TRPV2 receptors expression in both neurons and glia cell. With use of real-time (real-time PCR) we determined that the level of these genes expression in different neurons is identical. It was also shown that the level of mTRPVI in GABAergic neurons was considerably higher, than the level of mTRPV2, while expression of genes in the glutamatergic neurons did not differ. Findings about expression of TRPV 1 and TRPV2 allow to suppose that vanilloid receptors can play an important functional role in the hippocampal neurons. PMID- 18416179 TI - [The role of mitochondrial uncoupling proteins in the development of changes of endothelium-dependent reactions of the heart and vessels in experimental diabetes mellitus]. AB - The influence of mitochondrial uncoupling protein genipin on endothelium dependent reaction of vessels, heart contractility and myocardial oxygen consumption was studied at streptozotocin-induced rat model of diabetes mellitus. The partial restoration of damaged at diabetes mellitus vascular reactions as well as decrease ofmyocardial oxygen consumption has been shown after intraperitoneal injection of genipin. For example, after the introduction of 10 mg/kg genipin were shown a partial restoration of endothelium-dependent dilatation of aorta and coronary vessels, increase of contractive strength dependent responses of vascular smooth muscle, decrease of portal vein isolated strips stiffness, the improvement of contractive properties and decrease of myocardial stiffness. These data suggest a possible role for mitochondrial uncoupling protein in the development of changes of heart and vascular responses observed at experimental diabetes mellitus. PMID- 18416180 TI - [Systematic positioning errors and electromyographic activity of muscles in the human single-joint movements]. AB - Accuracy of positioning was studied on the elbow joint movements in healthy adult humans. Individual trials consisted of the conditioning and test movements (CM, TM) produced with visual guidance and without it, correspondingly. The two types of CMs included a phase of slow movement (flexion - P1, or extension - P2) to the same target position (50 degrees in the most experiments). P1 and P2 CMs started, correspondingly, from the full extension position (0 degrees), or after preliminary flexion (100 degrees). Subjects were asked to remember the target position during CMs and to reproduce it in TM. The experiments have shown that position errors depend on the type of CMs. The subjects overestimated the target position, the errors were more significant in TMs produced after P2 CMs as compared with P1 CMs. Full-wave rectified, averaged EMGs were compared for the two types of CMs. In flexors the dynamic phases ofEMG activity were statistically greater in P1 than in P2 CMs (a group of 25 subjects), while the stationary components of EMG did not differ. Possible mechanisms of the position errors are discussed. It is suggested that after-effects in the activity of muscle spindles may be the main source for the position errors. PMID- 18416181 TI - [The changes of expression and Fc-gamma-receptor's polypeptide composition of fetal small intestine enterocytes in Bos primigenius taurus L]. AB - The expression and polypeptide composition of Fc-gamma-receptors of enterocytes from Bos primigenius taurus L. intestine at 3-, 5-, 7-, 9- month of fetal development was investigated. The results of immunobloting show similar composition of Fc-gamma-receptors extracted from apical and basolateral membranes. The proteins that bind IgG after PAAG electrophoresis and transferring on nitrocellulose were observed as 120, 87, 72 and 43 kDa polypeptide line. The changes of each polypeptide contents were related to the changes of total content of Fc-gamma-receptor proteins. The rise in concentration of Fc-gamma-receptors on apical membrane was observed from 3-rd to 7-th month of fetal development. Maximal concentration of these proteins was detected on enterocytes at 7-th month of fetal development. Fc-gamma-receptors content on basolateral side was higher than on apical side. The presence of Fc-gamma-receptors on enterocyte's membrane indicates on active recycling of these receptors on plasma membrane and reflects early development of immune system in Bos primigenius taurus L. fetus. PMID- 18416182 TI - [Spatial distribution of synaptic vesicles in CA1 hippocampal synapses under conditions of induced long-term potentiation in vitro]. AB - Prolonged activation of excitatory glutamatergic synapses causes modifications in their functioning and ultrastructural organization. While postsynaptic activity induced changes have been relatively well studied, the data on spatial dynamics of synaptic vesicles (SV) under conditions of synaptic activation are still lacking. Using organotypic hippocampal slice cultures as a model system and electron microscopy as a technique, we analyzed changes in SV numbers and their spatial distribution in spine synapses ofhippocampal CA1 area. Two approaches were used to activate synapses: a protocol of brief oxygen-glucose deprivation known to induce so-called anoxia-hypoglycemic long-term potentiation (LTP), as well as high frequency stimulation of Schaffer collaterals inducing LTP of evoked postsynaptic potentials in CA1 synapses. Observations during the first hour after stimulation (30 and 60 min time-points) have shown that in both cases active functioning of synapses leaded to decrease in the total SV number as well as to depletion of the readily releasable SV pool. Both experimental protocols caused a decrease in spatial clustering of SV which was more pronounced after anoxia hypoglycemic LTP. Possible mechanisms and functional consequences of these phenomena are discussed. PMID- 18416183 TI - NADPH-diaphorase activity and neurovascular coupling in the rat cerebral cortex. AB - The distribution of NADPH-diaphorase-reactive (NADPH-dr) neurons and neuronal processes in the cerebral cortex and basal forebrain and their association with parenchymal vessels were studied in normal adult rats using NADPH-d histochemical protocol. The intensely stained cortical interneurons and reactive subcortically originating afferents, and stained microvessels were examined through a light microscope at law (x250) and high (x630) magnifications. NADPH-dr interneurons were concentrated in layers 2-6 of the M1 and M2 areas. However, clear predominance in their concentration (14 +/- 0.8 P < 0.05 per section) was found in layer 6. A mean number of labeled neurons in auditory (AuV), granular and agranular (GI, AIP) areas of the insular cortex was calculated to reach 12.3 +/- 0.7, 18.5 +/- 1.0 and 23.3 +/- 1.7 units per section, respectively (P < 0.05). The distinct apposition of labelled neurons to intracortical vessels was found in the M1, M2. The order of frequency of neurovascular coupling in different zones of the cerebral cortex was as following sequence: AuV (31.2%, n = 1040) > GI (18.0%, n = 640) > S1 (13.3%, n = 720) > M1 (6.3%, n = 1360). A large number of structural associations between labeled cells and vessels in the temporal and insular cortex indicate that NADPH-d-reactive interneurons can contribute to regulation of the cerebral regional blood flow in these areas. PMID- 18416184 TI - [The changes of free radical lipid peroxidation, antioxidant system activity, nitrogen oxide level in iodo-deficient hypothyrosis]. AB - The state of lipid peroxidation system, antioxidant defense enzyme activity, nitric oxide level during iodine defficiency hypothyrosis have been studied in the experiments on the laboratory rats. It has been established that in hypothyrosis the intensity of free-radical reactions rise greatly, nitric oxide level in blood serum decreases, antioxidant defense processes are depressed. PMID- 18416185 TI - [Connection of functional characteristics and ultrastructure of the lung and myocardium in hypoxia and srtess]. AB - The estimation of changes in breathing, circulation, gas exchange, lipid peroxidation, which can be connected with or determined by morphofunctional state of lung tissue and myocardium in rats under acute hypoxic hypoxia, acute dose hemorrhage and immobilization stress were made. Such approach is important not only from theoretical point of view, for the elucidation and definition of mechanisms which are responsible for function of oxygen-transport systems under unfavorable influences on the organism, but present particularly practical interest. The exposure of correlation between structure and function in investigated organs may permit to forecast the availability of some ultrastructural violations under ventilation or circulation changes in the organism. PMID- 18416186 TI - [Peculiarities of L-arginine metabolism in the blood leukocytes in experimental diabetes mellitus]. AB - In experimental streptozotocin-induced diabetes, the activity of enzymes which have common substrate (L-arginine) changes in leucocytes of peripheral blood. It leads to misbalance between different pathways of arginine metabolism: NO synthase (oxidative) pathway is activated, whereas arginase (nonoxidative) one is depressed. The injection of L-arginine under diabetes activated nonoxidative pathway, which can be seen in possible decrease in NO-synthase activity with unchanged arginase activity. After injection ofaminoguanidine in animals under diabetes the efficiency of both pathways in leucocytes decreases, which may be the basis of reconstitution of physiological pool to the relatively essential amino acid L-arginine. PMID- 18416187 TI - [Effect of selenium-containing supplements on the indices of specific immunity and nonspecific resistance in chicken]. AB - The deficit of selenium is related to immunity worsening. Selenium improves an immune answer and raises disease resistance. The aim of our work was to study the influence of organic selenium in comparison with inorganic selenium on the factors of unspecific resistance and state of humoral immunity of chicken broilers. PMID- 18416188 TI - [Effect of the diet with different microelement composition on the state of alveolar and pelvic bones in rats]. AB - During this experimental study with 6 week-old rats the rate of alveolar bone resorption increased with the balanced diet and drinking water containing 50 mg/l NaF, which is similar to the group of rats observed in case of periodontitis model with the same diet and supplemented with ammonium chloride 5 mg/kg. The mineral complex (Mg-sulfat, Zn-sulfat, Mr-sulfat, Na-citrat) 300 mg/kg possessing a pronounced anti acidosis effect when added to the same balanced diet and sodium fluoride in drinking water with the same concentration prevents resorption processes in the alveolar bone and reduces toxicity action of fluorides on the alveolar and pelvic bones by decreasing metabolic acidosis in the bone tissue. PMID- 18416189 TI - [Cross-spectral analysis of the stroke blood volume oscillations and R-R interval duration among men during different loadings]. AB - The correlation of the stroke blood volume oscillations and the R-R interval duration was investigated among 125 healthy young men at rest, with controlled breathing, at tilt test, with mental and physical loadings with the help of the cross-spectral analysis. Maximums and minimums of the cross-spectral power for these indexes were found to be groupped in three frequency ranges: 0-0.04 Hz, 0.07-0.13 Hz and 0.15-0.4 Hz. The positive tops of the cross-spectral power had the biggest amplitude in the 0-0.04 Hz, 0.07-0.13 Hz frequency ranges in the most conditions; negative tops stipulated with breathing--in the 0.15-0.4 Hz frequency ranges. The structure of the cross-periodgrams changed reliably with different loadings if compared with lying position at rest. The maximum top amplitude of the cross-periodgram was reproduced at the largest extent with repeated changes in 235 +/- 31 days in the ranges of 0.07-0.13 Hz, having median frequency of 0.09 0.01 Hz in all conditions. PMID- 18416190 TI - [Alimentary and oxygen deprivation as the modulator of the bone tissue physiological remodelling rate in young rats]. AB - Influencing of 28-days alimentary deprivation and intermittent hypoxia to 3 months Wistar male rats (n = 30) bone tissue physiological remodelling is studied. We investigated 3 groups of animals, I group was control, II--food limitation mode (-40% in relation to the normal ration) and III--animals, which breathed hypoxic gas mixture with 13% O2 in nitrogen (HGM) during 4 hours in the intermittent mode (10 min of deoxygenation and 10 min of reoxygenation). It is showed significant increasing of melatonin and glicozaminoglikans levels in the rats II and III gr. serum. Activity of alkaline phosphatase in the bone tissue of III gr. rats increased and in the bone tissue and serum of II gr. rats for 1.2 and 1.4 time accordingly. We registered significant increasing of serum acid phosphatase activity in animals, which have breathed hypoxic gas mixture. The IGF I gene expression level did not change practically in both experimental groups. We conclude, that alimentary deprivation and intermittent hypoxia have positive effects on the physiological remodelling of bone tissue. PMID- 18416191 TI - [Physiological properties and possible correction of adenosine triphosphate sensitive potassium channel function]. AB - Unique function of ATP-sensitive K channels (K(ATP)) is maintenance of interrelation between a metabolism and excitability of cells of various bodies and fabrics. In smooth muscle cells activation KP channels lead to vasodilatation, in beta-cells of a pancreas these channels play a role in interface of excitability of a membrane and secretion of insulin. Sulfonylurea drugs lowering a level of glucose in blood which use for treatment of a 2 type diabetes,--influence, contacting SUR 1 subunit and oppressing thus a K(ATP) current. It leads to membranes depolarisation, to Ca2+ entrance in cells and secretions of insulin. As opposed to this, diazoxide and pinacidil which reduce allocation of insulin, reduce arterial pressure, are specific K(ATP) activators. These preparations render therapeutic effects, stimulating K(ATP) and as consequence, limit increase of a level intracellular Ca2+ at depolarisation. At an ischemia of heart activation of K(ATP) channels reduces duration of potential of action, and it leads to reduction of Ca2+ entrance and renders cardio protective action. For last years significant successes in understanding of molecular mechanisms, determines K(ATP) channels activity, and in finding-out of their role in physiological and pathological processes are achieved. In this review of the literature the basic achievements in this area, and also problems which should be solved are considered. PMID- 18416192 TI - [Foreign accent syndrome in a case of dissociative (conversion) disorder]. AB - Psychiatric symptoms are often manifested in verbal expression. Generally, the contents of such expression are pathological. The formal abnormalities of speech are also observed in various mental disorders, as far as quantitative abnormalities are concerned. However, disturbance of intonation, namely disprosody, is more commonly observed in organic disorders of the brain. When the accent of words and the intonation of sentences changes from that of a native speaker, the speech sounds like the broken language of untrained foreigners. Such foreign accent syndrome is usually an issue of neuropsychology. In this paper, the authors report a case of foreign accent syndrome without organic brain syndrome. The patient was a 44-year-old woman, who developed panic disorder about year after her father's death. Then she developed aphonia. After aphonia was resolved, she began to speak haltingly as if a Chinese woman was trying to speak Japanese. Organic brain diseases were subsequently excluded. She had complicated familial conflicts, including a divorce from a violent and faithless husband, interpersonal difficulties with her husband's parents, and her pubertal daughter. We diagnosed her with dissociative (conversion) disorder of ICD-10. Our patient is clinically interesting, because case reports of dysprosody are unusual and often involve organic brain diseases. We suppose that foreign accent syndrome in our patient is a variant of aphonia, and the patient unconsciously assigned the symptom two ambivalent rolls: to snow that she cannot speak well, and to express her meaning. In addition, she had a Chinese-speaking aunt-in-law who was her ideal role model. We surmise that her symptom signifies identification with her aunt-in-law. PMID- 18416193 TI - [Novel assessment of intracellular calcium transient decay in cardiac muscle by curve-fitting with half-logistic function]. AB - A decrease in intracellular calcium (Ca2+) concentration in the cardiac muscle is one of the important factors to induce myocardial relaxation. A mono-exponential (m-E) function has been used for assessing myocardial relaxation curve of isometric tension and intracellular calcium transient (CaT) decay, and the m-E time constants for the relaxation curve of isometric tension (F tau E) and CaT decay (Ca tau E) have been recognised as lusitropic indices. However, we found that a half-logistic (h-L) function fits the relaxation curve of isometric tension much more precisely than the conventional m-E function in the ferret right ventricular (RV) papillary muscle. Moreover, we demonstrated that the goodness of the h-L fits for CaT decays was superior to the goodness of the m-E fits in the rabbit RV and murine left ventricular papillary muscles. The changes in the h-L time constants for the relaxation curves of isometric tension (F tau L) and CaT decays (Ca tau L) with the different onsets were significantly smaller than the changes in F tau E and Ca tau E, respectively. The differences in the h L non-zero asymptotes for the relaxation curves of isometric tension and CaT decays with the different onsets were smaller than the changes in the m-E non zero asymptotes. The h-L function model characterises the amplitudes and time courses of the relaxation curve of isometric tension and CaT decay more precisely than the m-E function model, and thus F tau L and Ca tau L serve as more novel and reliable lusitropic indices. Simultaneous analysis of myocardial relaxation curve of isometric tension and CaT decay using h-L functions can become a useful method for assessment of myocardial calcium handling. PMID- 18416194 TI - [Effects of postoperative continuous epidural analgesia after laparoscopy assisted colectomy]. AB - BACKGROUND: This study was conducted to evaluate the effects of continuous epidural analgesia on the postoperative pain and the early recovery after laparoscopy-assisted colectomy (LAC). METHODS: A total of 50 patients undergoing elective LAC were investigated for postoperative pain score and other variables retrospectively. RESULTS: Thirty patients who had received epidural analgesia (E group) showed significantly lower pain scores for 2 and 24 hours after surgery, compared with twenty patients who had received analgesics intramuscularly or transrectally (C group). The patients in the E group needed significantly less supplemental analgesics than those in C group for 12 to 24 hours after the operation. The incidence of postoperative complications and the early recovery were not significantly different between the two groups. CONCLUSIONS: These results show that postoperative continuous epidural analgesia is effective for postoperative pain relief in patients after laparoscopy-assisted colectomy. PMID- 18416195 TI - [Case of epidural hematoma developed after extraction of the epidural catheter and heparin injection in a patient with pulmonary embolism after colectomy]. AB - A 65-year-old man underwent transversal colectomy for colon cancer under combined epidural and general anesthesia. On the 1st postoperative day, he developed consciousness loss and low SpO2 (< 90%) after walking, and pulmonary embolism was diagnosed by CT-scan and pulmonary scintigraphy. His consciousness and hemodynamic state recovered, and anticoagulation therapy was started after extraction of the epidural catheter. Heparin 5000 units was injected and continuous injection was started. Five hours after the extraction of the catheter, he developed paraplegia and analgesia below L1, and epidural hematoma was found with magnetic resonance imaging (MRI). Emergent laminectomy was performed and the hematoma was removed. The day after laminectomy, injection of heparin was started and 1 g x day(-1) of methylpredonisolone administered for 3 days. His paraplegia did not improve after the laminectomy. We discussed about pulmonary embolism and epidural hematoma. PMID- 18416196 TI - [Use of high-dose dexmedetomidine infusion for anesthesia and sedation in a patient for microlaryngeal surgery maintained with spontaneous breathing]. AB - We experienced the anesthetic management using high-dose dexmedetomidine for microlaryngeal surgery maintaining spontaneous breathing. The anesthesia was maintained with dexmedetomidine infusion (initial dose 6 microg x kg(-1) x hr(-1) over 10 min followed by continuous infusion of 0.5 microg x kg(-1) x hr(-1)), intermittent small doses of fentanyl and topical application of lidocaine on the tongue, pharynx and larynx. The infusion of dexmedetomidine was increased over 30 min to 3 microg x kg(-1) x hr(-1) to reach the adequate sedation level and maintained at this rate for a further 15 min during the operation. During the whole perioperative period, there was no respiratory depression as measured by arterial blood gas analysis that recorded normal PaCO2 in the patient breathing supplemental oxygen. Hypotension (systemic arterial blood pressure less than 100 mmHg) occurred twice during dexmedetomidine administration, but was normolized by ephedrine administration. The preservation of respiratory drive offers the possibility that this anesthetic technique may be another method for providing anesthesia for the patient with a difficult airway. Moreover, there is one consensus on the importance of the basic principle that adequate topical or intravenous anesthesia is also essential during high-dose dexmedetomidine infusion. PMID- 18416197 TI - [Anesthetic management of coronary artery bypass grafting for unstable angina pectoris in a patient undergoing home oxygen therapy]. AB - A 70-year-old man with pneumoconiosis receiving home oxygen therapy presented with chest pain. Coronary angiogram showed severe stenosis of his left main trunks and required emergency off-pump coronary artery bypass grafting. Anesthesia was induced and maintained with propofol, fentanyl and vecuronium. The trachea was intubated and the lungs were ventilated with oxygen and air maintaining the PaO2 between 80-100 mmHg and the PaCO2 between 55-65 mmHg. The operation was performed uneventfully. In the intensive care unit, the trachea was extubated after ensuring adequate respiratory function. The postoperative course was satisfactory. PMID- 18416198 TI - [Hypoxemia during cesarean section due to massive pleural effusion in a patient with preeclampsia]. AB - We report hypoxemia during cesarean section in a 33-year-old woman with preeclampsia. She developed generalized body weight increase, severe proteinuria and decrease of platelets at week 33 of gestation. Cesarean section was scheduled under general anesthesia. The patient did not complain of dyspnea preoperatively. After induction of general anesthesia, arterial oxygen saturation decreased from 99 to 88%. Massive left pleural effusion was detected and chest drainage was performed, and oxygenation was improved. Accumulation of pleural effusion tends to be over-looked because many parturients with pleural effusion do not complain of dyspnea. Physicians should pay attention to pleural effusion in parturients, especially those with risk factors such as preeclampsia, generalized body weight increase and administration of beta stimulant. PMID- 18416199 TI - [Anesthesia in a patient with alkaptonuric ochronosis for total hip arthroplasty]. AB - Alkaptonuric ochronosis, caused by a deficiency of homogentisate 1,2-dioxygenase, is a rare, autosomal recessive, metabolic disorder. Accumulation of homogentisate acid (HGA) at the connective tissue destructs the spine and large joints, and cardiac valvular disease is prominent. In this report, we describe a case of alkaptonuric ochronosis for anesthetic management. A 75-year-old female patient with the disease was scheduled for a total-hip arthroplasty. We avoided applying general anesthesia for her valvular regurgitations. Spinal anesthesia was achieved successfully, and resulted in a hypesthesia level at T12. Although a epidural catheter was indwelled with no leak of cerebrospinal fluid, an accidental dural puncture appeared later during the surgery, suggesting a subdural catheterization. She had an uneventful perioperative course without any symptoms. In the patient of alkaptonuric ochronosis, the dura and arachnoid membrane could be damaged made vulnerable by HGA. In addition, since the clinical findings resemble ankylosing spondylitis, degenerative changes such as a narrowing of the disk space and spine fusion would make the regional technique unsuccessful. In term of anesthesia, alkaptonuric ochronosis requires ingenuity since there are a number of factors associated with prevention of untoward complications. Each case is to be evaluated individually and managed carefully. PMID- 18416200 TI - [Early goal-directed therapy (EDGT) using continuous central venous oxygen saturation monitoring in a patient with septic shock]. AB - Septic shock is an adverse clinical condition resulting in multiple organ failure from global tissue hypoxia. The importance of initial treatment is widely recognized. Thus, guidelines for septic shock recommend early goal-directed therapy (EGDT) during the first six hours of treatment. Central venous oxygen saturation monitoring is useful to maintain adequate tissue oxygen delivery. A newly developed central venous oximetry catheter (PreSep Oximetery Catheter, Edwards Lifesciences) allows continuous and easy monitoring of central venous oxygen saturation. This report shows the usefulness of this catheter in a patient who developed septic shock during an emergency operation for perforated bowel. By using EGDT perioperatively with continuous central venous oximetry, multiple organ failure might be successfully avoided. PMID- 18416201 TI - [Anesthesia for a patient with stiff-person syndrome]. AB - Stiff-person syndrome is a rare disease characterized by muscle rigidity and painful spasms in the axial and limb muscles. The authors reported here a case of an axilally lymphadenectomy in a 46-year-old woman with stiff-person syndrome. With train of four ratio (TOFR) monitoring at the ulnar nerve, general anesthesia was induced and maintained with fentanyl, vecuronium and propofol with target controlled infusion. A TOFR, BIS monitor and invasive arterial pressure monitoring were employed. During the operation, there was no muscle rigidity and spasm. Ten minutes after the operation, she was fully awake and train of four ratio recovered to 95%, and extubated uneventfully. We chose propofol, because of previous reports about prolonged hypotonicity by interaction of baclofen and isoflurane. Preoperative good symptom control, choice of total intravenous anesthesia (TIVA), and application of the electrical nerve stimulator prevented postoperative hypotonia and resulted in safe anesthetic management. PMID- 18416202 TI - [Case of postoperative paraplegia caused by idiopathic spinal cord infarction in a young male patient]. AB - We report a case of spinal cord infarction in a 20-year-old man, who underwent abdominal surgery under general anesthesia combined with epidural anesthesia. The patient was a healthy young man with no medical history. After the operation, he complained of weakness and sensory blockade of both legs. MRI revealed spinal cord infarction at the T9-10 vertebral levels. Steroid pulse therapy was introduced, and paraplegia almost disappeared. Spinal cord infarction in a young man is very rare, but should be considered in case of postoperative paraplegia even in a young patient. PMID- 18416203 TI - [Are transcutaneous blood gas measurement monitors (SenTec Digital Monitor system) useful for the respiratory management of chronic respiratory failure?]. AB - BACKGROUND: The level of arterial carbon dioxide partial tension (PaCO2) is a sine qua non for the determination of the adequacy of ventilation. The aim of this study was to assess the usefulness of SenTec Digital Monitor System in measuring transcutaneous arterial carbon dioxide partial tension (PCO2) non invasively for the management of chronic respiratory failure. METHODS: Three ICU patients suffering from chronic respiratory failure were enrolled. We recorded the data of PCO2 measured by a transcutaneous ear sensor (V-Sign : SenTec Inc.) and the data of PaCO2 (GASTAT-603ie Techno Medica Co., Ltd., Japan) obtained from arterial blood gas analysis. RESULTS: The mean PCO2 was 67.2 mmHg (min. 65.9 mmHg, max. 80.2 mmHg, n = 10), and the mean PaCO2 was 71.2 mmHg (min. 67.2 mmHg, max. 79.0 mmHg, n = 10). Regression analysis showed good correlation between PCO2 and PaCO2 (PCO2 = .95 x PaCO2-0.18 mmHg; R = 0.74). Bland-Altman analysis of PCO2 yields a bias of d = 3.9 mmHg with limits of agreement (1SD) -0.4 mmHg, +8.2 mmHg. CONCLUSIONS: SenTec Digital Monitor System enables non-invasive and reliable trend monitoring of PCO2 levels in patients with chronic respiratory failure. PMID- 18416204 TI - [Successful nasal intubation using airway scope with gum elastic bougie in a case of difficult airway]. AB - AWS (Airway Scope, Pentax, Tokyo), a new videolaryngoscope, was originally designed to facilitate oral laryngoscopy and tracheal intubation. We describe a successful case of nasal intubation with a combination of AWS and GEB (gum elastic bougie). A 50-year-old man with a past history of difficult intubation was scheduled to undergo subtotal esophagectomy. As the patient was scheduled to be ventilated overnight after the operation in the intensive care unit, nasal intubation with a combination of AWS and GEB was planed. Anesthesia was induced with intravenous propofol 100 mg, fentanyl 200 microg and vecuronium 7 mg and endotracheal tube was inserted through the nostril. Oral insertion of AWS showed full view of the glottis and nasal insertion of GEB was easily advanced into the trachea through an endotracheal tube, allowing tracheal intubation while utilizing GEB as a stylet. No complication occured in this case. In conclusion, nasal intubation using AWS combination with GEB is useful for patients with difficult airways and is easy to perform. PMID- 18416205 TI - [Case of serious lidocaine intoxication]. AB - Lidocaine is frequently used as a local anesthetic and has very low incidence of serious complications. I report on a case of lidocaine intoxication in a 66-year old man who presented with seizure and cardiac periarrest following the brachial plexus block with 450 mg of lidocaine. Twelve minutes after the lidocaine administration, he became unconscious and 29 minutes after, developed seizure. Bradycardia and subsequent cardiovascular depression were found 74 minutes after the lidocaine block. The patient was successfully treated with diazepam, atropine sulphate and transcutaneous pacing for each symptom, respectively. PMID- 18416206 TI - [Anesthetic management of a pediatric patient with Beckwith-Wiedemann syndrome accompanied by macroglossia]. AB - We report the anesthetic management of an infant with Beckwith-Wiedemann syndrome having massive macroglossia and umbilical hernia. Umbilical hernia repair and tongue reductions were performed under general anesthesia. Although a difficult airway was anticipated, we did not have difficulty in her airway management, even with tracheal intubation. After the operation, mechanical ventilation was required for three days because of airway obstruction by secretions in the mouth and her edematous tongue. She had no complications and the postoperative course was uneventful. PMID- 18416207 TI - [Anesthetic management of a case of craniotomy using TCI of remifentanil with intraoperative monitoring of motor evoked potential]. AB - We experienced anesthetic management using target-controlled infusion (TCI) of remifentanil for craniotomy in which monitoring of motor evoked potential (MEP) was performed. Anesthesia was induced and maintained by TCI of propofol and remifentanil. Vecuronium bromide was not administered except for facilitating tracheal intubation. Target effect-site concentrations (ESCs) of remifentanil during intubation, exposure, dura incision, microsurgery and closure were 6, 10, 8, 5 and 8 ng x ml(-1), respectively. Myogenic MEP was sufficiently elicited throughout the microsurgery without patient's body movement. Extubation was completed 10 min after the end of surgery with administration of remifentanil continued at a target ESC of 2 ng ml(-1) after surgery. Emergence from anesthesia was good without complaint of pain or respiratory disorder or new neurological deficit. It has been reported that remifentanil is suitable because of its wide dosage window with respect to recording MEP. ESC of remifentanil was fixed during continuous infusion, but its absolute value varies depending on lean body mass and/or age. The use of TCI enabled easy elimination of the above effects, adjustment of ESC to the expected value and maintenance of ESC of remifentanil at a constant level. TCI of remifentanil might be suitable for anesthesia with monitoring of MEP. PMID- 18416208 TI - [Heparin resistance associated with elevated factor VIII]. AB - An 84-year-old female patient was scheduled to undergo AVR, CABG, and Maze procedure. She had a history of hypertension, cerebral infarction, and branch retinal vein occlusion. Warfarin was administered preoperatively. Before the cardiopulmonary bypass (CPB), heparin 5,000 units was administered. Activated coagulation times (ACTs) before and after CPB were 123 sec and 157 sec, respectively. Additional heparin of 5,000 units extended ACT to 221 seconds, which was not enough for the CPB. Heparin 10,000 units was added, and ACT was 157 sec. AntithrombinIII (ATIII) and platelet counts were 75% and 270,000 mm(-3), respectively. ATIII 1,500 units was administered. ACT and ATIII became 133 sec and 123%, respectively. Because heparin resistance did not respond to ATIII, the operative method was changed to off-pump CABG. A postoperative examination revealed high factor VIII activity of 263%. Other results were as follows: protein C antigen, 40%; protein S antigen, 65%; factor VII, 50%; platelet factor 4, 12%; heparin cofactor II, 104%; von Willebrand factor antigen, 181%; heparin PF4-IgG antibody, negative; factor VIII inhibitor, negative. The low values of protein C, protein S, and factor VII may have been caused by warfarin. Other values were normal, except for the von Willebrand factor antigen. PMID- 18416209 TI - [Massive subcutaneous emphysema following percutaneous tracheostomy]. AB - Although percutaneous dilatational tracheostomy (PDT) is fast becoming the method of choice for securing an airway in chronic ventilated patients in an intensive care unit (ICU). Subcutaneous emphysema is an unusual and sometimes lethal complication which may extend the length of stay in the ICU. We report a case of massive subcutaneous emphysema without tracheal wall laceration that occurred in the ICU after PDT. An 81-year-old woman was admitted to our ICU due to infective exacerbation of chronic obstructive airways disease. Her medical therapy included nebulized bronchodilators, steroids and empirical antibiotics. Within thirty-six hours of initial endotracheal intubation PDT was performed. Several hours following the procedure, the patient developed massive subcutaneous emphysema encompassing her entire body. No pneumothorax was identified on subsequent chest X-ray. Laryngoscopic and bronchoscopic examination showed no evidence of tracheal wall laceration. We discuss the etiology and management plan for this rare complication. PMID- 18416210 TI - [Successful anesthetic management of two high-risk elderly patients using propofol-remifentanil]. AB - This report deals with anesthetic management using remifentanil in high-risk elderly patients with many complications such as arteriosclerosis and obstructive lung disease other than the disease which was the object of operation. Case 1; an 87-year-old woman with complete left bundle branch block and aortic valve stenosis was scheduled for cholecystectomy, intestinal tract incision and gallstone extraction. Case 2; a 79-year-old woman with obstructive lung disease was scheduled for sigmoidectomy and cholecystectomy. The use of remifentanil in anesthetic management to control the blood pressure and the pulse, was useful. They were extubated within 30 minutes after the operation. They received epidural anesthesia 30 minutes before the remifentanil discontinuance. After remifentanil is discontinued, they had little pain. Elderly patients considered at high-risk were treated under total intravenous anesthesia with remifentanil and propofol. PMID- 18416211 TI - [Case of severe transfusion-related acute lung injury during coronary artery bypass surgery]. AB - We report a case of transfusion-related acute lung injury (TRALI) during coronary artery bypass surgery. A 73-year-old man developed severe hypoxemia after transfusion of platelet concentrate just before completion of surgery. The possibility of cardiogenic pulmonary edema was excluded by transesophageal echocardiography. A chest X-ray showed bilateral pulmonary edema, suggesting TRALI. Steroid therapy and administration of sivelestat sodium hydrate and vasopressors were started. The patient's PaO2 /FIO2 ratio increased from 56 to 223 within 12 hrs. Anti-HLA and antigranulocyte antibodies were detected in the donated blood products. We assume that the cause of this case was an immune reaction through anti-HLA and/or antigranulocyte antibodies. TRALI is a potentially life-threatening, underrecognized and under-reported complication of transfusion. A better understanding and awareness is needed for medical staffs. PMID- 18416212 TI - [Drug errors during anesthesia]. AB - We report drug errors during anesthesia. The data were retrieved from the incident reports in a period of June 2000 to June 2003 at the Department of Anesthesiology, Fukuoka University Hospital. Drug errors occurred in 18 (0.12%) of 15,271 patients who were managed by anesthesiologists during the period. Among the 18 events, most frequent errors were 12 events of administration of wrong drugs. Errors in dosage were five, and wrong route of administration in one. There were no serious sequelae in the 18 patients. PMID- 18416213 TI - [Present state of anesthetic management for awake craniotomy in Japan]. AB - To avoid the neurological deficits after neurosurgical procedures, awake craniotomy applying intraoperative awake functional brain mapping has been employed. Anesthesia for awake craniotomy requires particular attention to airway management, control of seizures and measures for decreasing the anxiety of the patients. We investigated the current status of anesthetic management for awake craniotomy in Japan to establish a standard procedure for safe anesthesia. A questionnaire was sent to 80 universities with departments of anesthesia in Japan and 34 (43%) responded. In 19 institutes, awake craniotomies are being practiced. The first experience of awake craniotomy was in 1996 in Japan and since then most of the institutes have experienced only three or fewer cases. Airway management, control of nausea and vomiting, stable awakening during functional mapping and control of seizures were pointed out as problems during awake craniotomy. Based on the present results, our experience and the information from previous investigations, standard anesthetic management for awake craniotomy in our country will be documented. PMID- 18416214 TI - [Wastage of anesthetic related drugs in a university hospital]. AB - BACKGROUND: The cost of wasted anesthetic related agents has not been clear in Japanese hospitals. We investigated whether the trainees in anesthesiology influence the cost of wasted anesthetic related agents. METHODS: Investigation was carried out at the University of Tokyo Hospital. We interviewed each trainee in anesthesiology about all prepared anesthetic drugs and wasted ones at the end of each anesthetic management. RESULTS: The percentage of wasted ampoules of anesthetic related agents was 15.85%, but the percentage of wasted cost was 5.15%. A large difference was not observed in transition of training period, and this percentage was not improved by training. We considered that this wasted cost is within permissible ranges in comparison with other reports. CONCLUSIONS: During the training it is also important to develop a sense of medical economics. PMID- 18416215 TI - Establishing an association between a peri-operative perfusion score system (PerfSCORE) and post-operative patient morbidity/mortality during CPB cardiac surgery. AB - BACKGROUND: To date, there is no quality assurance program that correlates patient outcome to perfusion service provided during cardiopulmonary bypass (CPB). A score was devised, incorporating objective parameters that would reflect the likelihood to influence patient outcome. The purpose was to create a new method for evaluating the quality of care the perfusionist provides during CPB procedures and to deduce whether it predicts patient morbidity and mortality. METHODS: We analysed 295 consecutive elective patients. We chose 10 parameters: fluid balance, blood transfused, Hct, ACT, PaO2, PaCO2, pH, BE, potassium and CPB time. Distribution analysis was performed using the Shapiro-Wilcoxon test. This made up the PerfSCORE and we tried to find a correlation to mortality rate, patient stay in the ICU and length of mechanical ventilation. Univariate analysis (UA) using linear regression was established for each parameter. Statistical significance was established when p < 0.05. Multivariate analysis (MA) was performed with the same parameters. RESULTS: The mean age was 63.8 +/- 12.6 years with 70% males. There were 180 CABG, 88 valves, and 27 combined CABG/valve procedures. The PerfSCORE of 6.6 +/- 2.4 (0-20), mortality of 2.7% (8/295), CPB time 100 +/- 41 min (19-313), ICU stay 52 +/- 62 hrs (7-564) and mechanical ventilation of 10.5 +/- 14.8 hrs (0-564) was calculated. CPB time, fluid balance, PaO2, PerfSCORE and blood transfused were significantly correlated to mortality (UA, p < 0.05). Also, CPB time, blood transfused and PaO2 were parameters predicting mortality (MA, p < 0.01). Only pH was significantly correlated for predicting ICU stay (UA). Ultrafiltration (UF) and CPB time were significantly correlated (UA, p < 0.01) while UF (p < 0.05) was the only parameter predicting mechanical ventilation duration (MA). CONCLUSIONS: CPB time, blood transfused and PaO2 are independent risk factors of mortality. Fluid balance, blood transfusion, PaO2, PerfSCORE and CPB time are independent parameters for predicting morbidity. PerfSCORE is a quality of perfusion measure that objectively quantifies perfusion performance. PMID- 18416216 TI - Reduction of hemodilution in small adults undergoing open heart surgery: a prospective, randomized trial. AB - BACKGROUND: Given that there is an association between the degree of hemodilution during cardiopulmonary bypass (CPB) and postoperative complications, patients outcome might be improved if the nadir hematocrit concentration is kept within an optimal range. Smaller patients are more likely to have a low hematocrit during CPB: this phenomenon may be related, at least partially, to the extreme hemodilution induced by a large fixed CPB priming volume. METHODS: Forty patients with a body surface area (BSA) < 1.7 m2 undergoing open heart operations were randomized to either standard CPB with full prime volume (control group) or reduced prime extracorporeal circuit and vacuum-assisted venous drainage (VAVD) (study group). RESULTS: There were no significant differences between the groups with respect to baseline characteristics, body surface area, hematologic profile and operative data. Clinical outcomes were similar. Nadir hematocrit and hemoglobin on bypass were significantly lower in the control group (22 +/- 2.3 vs. 24 +/- 2.5%, p < 0.02 and 7.4 +/- 0.7 vs. 8 +/- 0.9 g/dl, p < 0.04, respectively). Postoperative chest tube drainage was significantly higher in the control group (272 +/- 253 vs. 139 +/- 84 ml, p < 0.04). There was no difference in blood transfusion in the two groups (0.5 +/- 1.14 vs. 1.0 +/- 1.77 units of packed red blood cells (PRBC), p = 0.29). CONCLUSIONS: Lowering CPB priming volume by means of using a small oxygenator and vacuum-assisted venous drainage (VAVD) resulted in a significant decrease of intraoperative hemodilution. This technique should be strongly considered for patients with a small BSA (<1.7 m2) undergoing open heart surgery. PMID- 18416217 TI - Evaluation of oxygenators and centrifugal pumps for long-term pediatric extracorporeal membrane oxygenation. AB - OBJECTIVE: Two extracorporeal membrane oxygenation (ECMO) circuits for children under 10 kg were evaluated and compared for plasma leakage, hemolysis, blood transfusions, and durability. METHODS: Group A (n=20) was supported by ECMO circuits with the Minimax oxygenator and the Biomedicus centrifugal pump. Group B (n=10) was supported by ECMO circuits with the Lilliput 2 ECMO oxygenator and the Rotaflow centrifugal pump. RESULTS: ECMO circuit durability, as measured by oxygenator lifespan, was significantly better in Group B than in Group A (p = 0.04). There was significantly lower hemolysis, measured by plasma free hemoglobin, in Group B (p = 0.019), and patients in Group B had significantly less need for antithrombin III transfusion (p = 0.004). No plasma leakage was observed in Group B oxygenators, but plasma leakage was observed in all Group A oxygenators. CONCLUSION: The combination of a Rotaflow centrifugal pump and Lilliput 2 ECMO oxygenator in pediatric ECMO circuits improved durability and reduced circuit-induced hemolysis. This improvement may be due to the low priming volume, the oxygenator's plasma leakage resistance, the suspended rotor of the centrifugal pump, or a combination of these factors. PMID- 18416218 TI - 180 ml and less: cardiopulmonary bypass techniques to minimize hemodilution for neonates and small infants. AB - OBJECTIVE: To determine the efficacy of decreasing cardiopulmonary bypass (CPB) prime volume for neonates and small infants by using low prime oxygenators, small diameter polyvinyl chloride (PVC) tubing and removing the arterial line filter (ALF) in an effort to reduce intraoperative exposure to multiple units of packed red blood cells (PRBC). METHODS: Two retrospective database studies comparing neonatal CPB prime volume were undertaken: Study 1--A CPB circuit consisting of a 1/8 inch arterial line, a 3/16 inch venous line and a low prime oxygenator with 172 ml total circuit prime (n=74) was compared to a circuit with a 3/16 inch arterial line, a 1/4 inch venous line and a higher prime oxygenator with a 350 ml total circuit prime (n=74). Study 2--The 172 ml circuit (n=389) was compared to a circuit that included an ALF and had a total circuit prime volume of 218 ml (n=389). RESULTS: Study 1--of the 74 neonates and small infants whose CPB prime volume was 350 ml, 19 were exposed to two or more intraoperative exogenous PRBC units while only 3 neonates and small infants in the 172 ml prime group (n=74) received two or more units (p = 0.0002). Study 2--of the 389 neonates and small infants where an ALF was used (prime volume 218 ml), 54 were exposed to two or more exogenous PRBC units while only 36 of the 389 patients where an ALF was not used (prime volume 172 ml) received two or more units of intraoperative PRBCs (p = 0.0436). CONCLUSION: Decreasing the neonatal and small infant extracorporeal circuit prime volume by as little as 46 ml resulted in significantly fewer multiple exposures to exogenous PRBC units. PMID- 18416219 TI - Low molecular starch versus gelatin plasma expander during CPB: does it make a difference? AB - BACKGROUND: Non-protein plasma expanders carry a risk of potentially severe allergic reactions. As prime for cardiopulmonary bypass, we routinely use a gelatin plasma expander. Plasma expanding during anesthesia is achieved with high molecular starch (200/0.5 kDalton) in combination with Ringer Lactate solution (RL) and in the Intensive Care Unit (ICU) with a low molecular starch (130/0.4 kDalton). We evaluated the feasibility of low molecular starch in combination with RL (group LMSRL) versus gelatin plasma expanding (group GPE) for priming CPB circuits in patients undergoing cardiac surgery in a randomized prospective trial. METHODS: One hundred and eighty adults who underwent primary valve or coronary artery bypass graft (CABG) surgery were equally stratified into 3 series of 60 patients with the routinely used oxygenators; Capiox RX-25, CML Duo and Quadrox-D. Then they were randomised by drawing lots and allocated into the LMSRL or GPE groups. We compared hematocrit, hemoglobin, platelet count, activated clotting time (ACT), lactate and colloid osmotic pressure (COP), blood loss, transfusion need, urine production and ICU stay. In addition, we monitored the average trans-oxygenator fluid resistance (AFR) for each type of oxygenator. RESULTS: The COP is significantly lower in the LMSRL group (20mmHg +/- 0.2 versus 18 mmHg +/- 0.2, p < 0.0001); as was the total use of plasma expanders (3846 ml +/- 98 versus 3059 ml +/- 77, p < 0.001). All other parameters were not significantly different. When comparing the observed AFR for the three types of oxygenators, a lower AFR in the LMSRL group (p < 0.02) was noted for the Capiox RX-25. CONCLUSIONS: This study shows a lower need for plasma expanders in patients who receive only starch plasma expanders. Further, we noted a lower COP in the LMSRL group, but since the mean COP was >17 +/- 0.2 mmHg, this cannot be considered of clinical importance. In conclusion, our study result supports the use of low molecular starch as a good alternative choice for priming CPB. PMID- 18416220 TI - Effects of zero-balanced ultrafiltration on procalcitonin and respiratory function after cardiopulmonary bypass. AB - The abnormal conditions to which blood is subjected during cardiopulmonary bypass (CPB) trigger an activation of the inflammatory response and cause pulmonary dysfunction. It has been suggested that high-volume, zero-balanced ultrafiltration (ZBUF) facilitates clearance of inflammatory mediators and improves post-operative pulmonary function. Procalcitonin, a newly discovered inflammatory mediator, has been found to be increased after CPB and has been proven to be an appropriate parameter for predicting pulmonary dysfunction secondary to CPB. The aim of this study was to investigate the effects of zero balanced ultrafiltration (ZBUF) on procalcitonin (PCT) and respiratory function of infants with Tetralogy of Fallot (TOF) after CPB. Twenty infants with TOF undergoing open-heart total surgical correction were randomly assigned to two groups. The trial group was given ZBUF (50 ml/kg) and conventional ultrafiltration (CUF), while the control group was given CUF only. Plasma PCT and pulmonary function were monitored and compared between the two groups before the operation (T1), before rewarming (T2), at the end of the operation (T3), and at 12 h, 24 h and 48 h after the operation (T4-T6). PCT was decreased in the trial group between 12 h and 48 h post-operatively, but the differences did not reach statistical significance. The trial group's pulmonary compliance was higher at 12 h post-operatively (p < 0.05). Oxygenation index was increased in the trial group at the end of the operation and 12 h post-operatively (p > 0.05). Intubation time was shorter in the trial group (P < 0.01). A positive correlation was found between peak PCT concentration and intubation time. ZBUF appeared to improve ventilation and shorten intubation time. The improved respiratory function may be due to the lower plasma PCT. PMID- 18416221 TI - Influence of early re-infusion of autologous shed mediastinal blood on clinical outcome after cardiac surgery. AB - Various strategies have been proposed to decrease allogeneic blood transfusion requirements after cardiac surgery. The aim of the study was to evaluate the efficacy of collected and re-infused autologous shed mediastinal blood on a patient's postoperative course. Ninety patients who underwent heart surgery with cardiopulmonary bypass (CPB) were studied. The patients were divided into two groups: Group 1 (n=41) received the centrifuged autologous shed mediastinal blood collected from the cardiotomy reservoir 4 hours after surgery; in Group 2 (n=49) all shed mediastinal blood was discarded (control group). Haemoglobin (Hb), haematocrit (Hct), C-reactive protein values, and leucocyte count were compared before surgery, at 4 h and 20 h after surgery, and on the fifth postoperative day. We have measured serum procalcitonin (PCT) concentration at 4 h and 20 h after CPB. We assessed drained blood loss within 20 postoperative hours. Leucocyte count, Hb, Hct values, C-reactive protein, and procalcitonin concentration did not differ between the groups before and at 4 h after surgery. Hb, Hct level, and leucocyte count were similar at 20 hours and on the fifth day after surgery. At 20 hours after surgery, an increase of serum PCT concentration (>0.5-2 ng/mL) was more frequent in Group 2 (58.3% vs. 33.3%; p = 0.03). On the fifth postoperative day, C-reactive protein concentration was lower in Group 1 (71.74 +/- 15.23; p <0.01) compared to Group 2 (93.53 +/- 20.3). Postoperative blood loss did not differ between the groups. Requirement for allogeneic blood transfusion was significantly lower in Group 1 (14.6% vs. 38.8%; p < 0.02). Patients in Group 1 developed less infective complications compared with Group 2 (2.4% and 16.3%, respectively; p < 0.05). The length of postoperative in-hospital stay was shorter in Group 1 compared with Group 2 (9.32 +/- 2.55 and 16.45 +/- 6.5, respectively; p < 0.05). We conclude that postoperative re-infusion of autologous red blood cells processed from shed mediastinal blood did not increase bleeding tendency and systemic inflammatory response and was effective in reducing the requirement for allogeneic transfusion, the rate of infective complications and the length of postoperative in-hospital stay. PMID- 18416222 TI - Mathematical modeling of buffers used in myocardial preservation. AB - OBJECTIVE: Buffers added to myocardial preservation solutions are considered to be critical for resisting myocardium pH changes from the accumulation of protons (H+). Our hypothesis is that mathematical modeling of three clinically used buffers will define their individual buffering capacities under simulated clinical conditions. METHODS: The buffers, tromethamine (THAM), sodium bicarbonate (HCO3-), and L-histidine, were compared in terms of their buffering capacity (beta) under specific temperatures and concentrations, using a mathematical model. RESULTS: At 37 degrees C, the maximal beta (betamax) occurred at pH 7.75 for THAM, pH 6.10 for HCO3-, and pH 5.89 for L-histidine at equimolar concentrations. A decrease in temperature moved betamax to a higher pH value for each buffer. At clinical concentrations, L-histidine provided the greatest buffering capacity followed by HCO3- and THAM, respectively. DISCUSSION: This model permitted comparison of the above buffers under simulated clinical conditions. The assumption was that the magnitude of betamax at a given temperature determines which buffer(s) could be most effective for myocardial preservation. Also, the assumption was taken that these buffers are used in a closed system--where there is no continuous blood flow--and that the buffering ability of THAM and L-histidine were not influenced by the accumulation of CO2 as is HCO3-. THAM and L-histidine were more effective at hypothermic temperatures compared with HCO3-; however, HCO3- provided buffering at normothermic temperatures. Through the theoretical considerations of this study, we propose that combining HCO3- with THAM or L-histidine could be most efficacious for myocardial preservation during open heart surgery or organ transplantation. PMID- 18416223 TI - Thoraco-abdominal aneurysm repair in a Jehovah's Witness: maximising blood conservation. AB - Thoraco-abdominal aneurysm repair usually necessitates blood or blood product transfusion which is prohibited in Jehovah's Witnesses. We report the blood conservation strategy used during thoraco-abdominal aneurysm repair in a Jehovah's Witness. This included pre-operative recombinant erythropoietin, per operative acute normovolaemic haemodilution, cell salvage, aprotinin, restricted heparinisation, left atrial-distal bypass and recombinant factor VIIa. Post operative haemoglobin levels were maintained, but a left haemothorax necessitated re-thoracotomy on post-operative day 4. Following re-thoracotomy, Hb was 12.0 g.dL(-1) and platelet count 49 x 10(9).L(-1). Recombinant erythropoietin was recommenced. At discharge (day 12), Hb was 10.1 g.d(L-1). The patient remains well at one year. A thoroughly, pre-planned multi-disciplinary blood conservation strategy can be used to undertake high-risk procedures. PMID- 18416224 TI - Emergency operation on the dissected ascending aorta of an adolescent with aortic coarctation. AB - This case report describes the successful repair of an acute dissection of the ascending aorta in a 16-year-old girl. The dissection appeared to be related to the presence of an aortic coarctation. Furthermore, a bicuspid aortic valve was found. Echocardiography was helpful for quick pre-operative diagnosis. A computed tomography (CT) scan was used post-operatively to confirm these results. At the onset of extracorporeal circulation, active drainage was applied at the femoral vein by means of a routinely built-in shunt bypassing the venous reservoir. The practical implementation of this shunt is described in this report. One femoral artery and one femoral vein were cannulated, resulting in a blood flow index of 2.21.min(-l).m(-2) which proved to be sufficient for blood supply to both the lower and the upper parts of the body. Successful percutaneous dilatation of the coarctation was performed five months after the first operation. PMID- 18416225 TI - AvMA's silver wedding, and a funeral. PMID- 18416226 TI - Show me the money: the new death investigation system. PMID- 18416227 TI - Anecdotes, data and the curse of the media case study. PMID- 18416228 TI - Acrimonious contact disputes and so-called parental alienation syndrome: a model of understanding to assist with resolution. PMID- 18416229 TI - I see you. As focus sharpens on the intensive care unit, remote monitoring capabilities and integration issues are under the microscope. PMID- 18416230 TI - Capital control. Project management applications can help ensure massive construction endeavors don't spin out of financial control. PMID- 18416231 TI - Corralling documents. With the lines blurring between DMI and EMR software, how do CIOs know which solution is right for them? PMID- 18416232 TI - Sound wireless strategy. At St Luke's, an expanding wireless strategy always includes protecting the integrity of the network. PMID- 18416233 TI - Tapping imaging reports. A new set of standards looks to bring usability to imaging management reports, by letting clinicians mine the data they hold. PMID- 18416234 TI - Setting the standards stage. If health information exchange is to get to the next level, formulating standards will be an absolute key. PMID- 18416235 TI - Researching integration. As academic medical centers push the envelope of patient care IT, it's important for CIOs to keep applications up to speed. PMID- 18416236 TI - Banking on donations. With revenues and profits ever shrinking, philanthropy can go a long way to bolstering hospitals' bottom lines. PMID- 18416237 TI - Credit crunch hits hospitals. For healthcare organizations with less-than-stellar credit ratings, raising cash has become a whole lot tougher. PMID- 18416238 TI - Regs change roles. Potential expansion in health savings accounts puts onus on hospitals to collect upfront. PMID- 18416239 TI - Backbone of an EMR. The central role of pharmacy information systems comes to light in enterprise-wide IT projects. PMID- 18416241 TI - What if? Planning for potential disasters strains technology budgets and stretches overworked staff members. PMID- 18416240 TI - Adoption in its own time. Forget about pushing physicians to adopt IT, a number of external factors will ensure technology is eventually embraced. PMID- 18416242 TI - [Aedes albopictus, vector of chikungunya and dengue viruses in Reunion Island: biology and control]. AB - Chikungunya virus (CHIKV) and dengue virus (DENV) are mosquito-borne viruses transmitted by the Aedes genus. Dengue is considered as the most important arbovirus disease throughout the World. Chikungunya, known from epidemics in continental Africa and Asia, has up to now been poorly studied. It has been recently responsible for the severe 2004-2007 epidemic reported in the Indian Ocean (IO), which has caused several serious health and economic problems. This unprecedented epidemic of the IO has shown severe health troubles with morbidity and death associated, which had never been observed before. The two major vectors of those arboviruses in the IO area are Aedes aegypti and Aedes albopictus. The latest is considered as the main vector in most of the islands of the area, especially in Reunion Island. Ae. albopictus showed strong ecological plasticity. Small disposable containers were the principal urban breeding sites, and preferred natural developmental sites were bamboo stumps and rock holes in peri urban and gully areas. The virus has been isolated from field collected Ae. albopictus females, and in two out of 500 pools of larvae, demonstrating vertical transmission. Experimental works showed that both Ae. albopictus and Ae. aegypti from west IO islands are efficient vectors of dengue and chikungunya viruses. Since 2006 and all along the epidemic of CHIKV, measures for the control of larvae (temephos then Bacillus thuringiensis) and adults (fenitrothion, then deltamethrine) of Ae. albopictus where applied along with individual and collective actions (by the use of repellents, and removal of breeding sites around houses) in Reunion Island. In order to prevent such epidemics, a preventive plan for arboviruses upsurge is ongoing processed. This plan would allow a quicker response to the threat and adapt it according to the virus and its specific vector. PMID- 18416243 TI - [Phlebotomine sandflies from Madagascar (Diptera: Psychodidae). VI--A new subgenus (Vattieromyia) including three new species: Sergentomyia (V.) sclerosiphon, S. (V.) namo and S. (V.) anka]. AB - Three new species of Phlebotomine sandflies belonging to the genus Sergentomyia Franca & Parrot are described from Madagascar: Sergentomyia sclerosiphon n. sp. S. namo n. sp. and S. anka n. sp. The taxa are created on the basis of the morphology of females. The males of S. sclerosiphon n. sp. and of S. anka n. sp. have been associated to the females according to the homologies of a fragment of the mtDNA cytochrome b sequences. The original morphology of the spermathecae and the sclerification of a part of the individual spermathecal ducts allow the creation of a new subgenus, Vattieromyia subg. nov., which includes these three new species. PMID- 18416244 TI - [Ecology of Stomoxys flies (Diptera: Muscidae) in Gabon. I. First survey in different ecological areas]. AB - The stomoxyine flies are hematophagous diptera and potential vectors of various pathogenic agents. Like those of the Afrotropical Region, the stomoxyine flies of Gabon remain nearly unknown. For these reasons, an entomological survey was conducted in a transverse way in eight localities representative of the various ecological zones of Gabon. The survey was based on the use of Vavoua traps. Various environmental factors able to influence the captures were noticed and included into a canonical correspondence analysis. In total, 15,966 Stomoxys spp., belonging to seven species or subspecies, were captured. The apparent densities (DAP) expressed as the number of flies per trap and per day, were highest in Franceville (41), Bakoumba (40), Makokou (25) and Mouila (21). The most abundant species were S. n niger (33.4%), S. transvittatus (33%), then S. calcitrans (17%). The principal factors that could explain the variability of the captures were the degree of anthropisation, the botanical facies (savanna or forest), the presence of wild and domestic fauna and the nature of the vegetal cover of the ground. S. calcitrans, S. niger niger were abundant in the areas where human presence was manifest. S. xanthomelas was present in forest belts. S. transvittatus, S. omega, S. inornatus were ubiquitous species. S. niger bilineatus was found in savannas areas. PMID- 18416245 TI - [Fleas community in introduced Siberian chipmunks (Tamias sibiricus Laxmann) in Forest of Senart, France]. AB - We examined the fleas community in an introduced population of Siberian chipmunks, Tamias sibiricus, between 2005 and 2007, in the Forest of Senart (Essonne, France). We collected and identified 383 fleas on 463 chipmunks (total: 1,891 captures on 471 chipmunks). In 2005, 120 fleas were also collected on 65 bank voles, Clethrionomys glareolus, and on 25 wood mice, Apodemus sylvaticus, trapped within the same area. Ceratophyllus sciurorum sciurorum formed 73.6% of the chipmunks' flea community, with an annual prevalence (P) ranging between 8 and 13% and a mean intensity (I) ranging between 1.1 and 1.6 fleas per individual. Among the six other species infecting this Sciurid, Ctenophthalmus agyrtes impavidus constituted 17.2% (P: 1.6-2.2%; I: 1.1-2.6), and Megabothris turbidus 8.1% (P: 0.8-1.9%; I: 1.0-1.4) of the flea community, respectively. These last two species represented respectively 60.8% and 36.6% of the flea community on the bank vole and the wood mouse. Originated from Asia, chipmunks did not import any flea species to Senart, probably because they were used as pets before their release in the wild. Abundance in C. s. sciurorum increased with adult chipmunk density and with juvenile density in summer. On adult chipmunks C. s. sciurorum tented to decrease with increasing abundance of red squirrels (Sciurus vulgaris]. Moreover, the two other flea species mainly infected young chipmunks during the fall, and their number was not related to chipmunk density. However, the distribution of species within the flea community became more balanced with increase juvenile chipmunk density. Overall, these results indicate that the close phyletic relationship between chipmunks and red squirrels contributed in the acquisition and the spread of fleas by chipmunks. Primary and secondary hosts densities, their habitat use, and more specifically burrowing activities and tree canopy use, also played a role in the spread of fleas on chipmunks. PMID- 18416246 TI - Skrjabinelazia rizzoi n. sp. (Nematoda: Seuratoidea) from a Sicilian lacertid, with comments on specific and biological diversity in the genus. AB - Skrjabinelazia rizzoi n. sp. (Seuratoidea), from Podarcis sicula captured at Cammarata, Agrigento Province, Sicily, is distinct from the 13 known species of the genus, including Skrjabinelazio sp. Rizzo from Catania. It is identified with the following set of characters: in the male, short spicules and gubernaculum, thin body; in the female, buccal cavity with a crown of leaflets, body cuticle without internal ornamentation, presence of cephalic and caudal vesicles, tail with a terminal digitiform spike 42-48 microm long. The morphology of the new species supports our previous hypothesis of two Skrjabinelazia lineages, one with spicules and one without spicules, respectively linked to Lacertidae and Gekkonidae. In Palearctic lacertids, five named species are presently known, S. taurica from Crimea (Ukraine), S. hoffmanni from Beijing (China), S. pyrenaica from Pyrenees (Spain), S. vozae from Cevennes (France), S. rizzoi from Agrigento Province, Sicily (Italy), but analysis of some published works suggests a greater diversity. S. rizzoi infection, found in April-May in 1/5 lizards, was recent with young females in the host's stomach and intestine, and males in the stomach. One female contained four membranous-shelled eggs. The two other females contained a few hatched infective larvae, membranous-shelled eggs with developing embryos and, unexpectedly at this early stage, a few thick-shelled divided eggs. As in several other Skrjabinelazia species, the progeny of S. rizzoi are adapted for intra-host suprainfection and inter-host transmission, but in this species the production of resistant eggs appears in very young females. PMID- 18416247 TI - Hypobaric hypoxia-related impairment of pulmonary surfactant proteins A and D did not favour Pneumocystis carinii Frenkel 1999 growth in non-immunocompromised rats. AB - It has been suggested that patients with pulmonary surfactant impairment are more susceptible to Pneumocystis infection than healthy controls. Owing the fact that most patients with pulmonary surfactant impairment also suffer from hypoxia, we explored the effect of intermittent hypobaric hypoxia conditions on the ability of non-immunocompromised rats infected by endotracheal route with P. carinii to clear the infection from their lungs. Control rats, inoculated or not with P. carinii, were maintained in normobaric normoxic conditions, and were submitted or not to dexamethasone administration. It was found that even if hypobaric hypoxia weakened host immune mechanisms and impaired significantly the surfactant composition, mainly of surfactant proteins A and D, these changes were not enough to favour the Pneumocystis growth or to inhibit the clearing of Pneumocystis organisms from the lungs of non-immunocompromised rats. The potential influence of surfactant protein changes on Pneumocystis infection is discussed. PMID- 18416248 TI - Molecular differentiation of Entamoeba histolytica and Entamoeba dispar from Tunisian food handlers with amoeba infection initially diagnosed by microscopy. AB - The purpose of the study was to obtain more reliable epidemiological data concerning Entamoeba (E.) histolytica infection in Tunisian food handlers using established molecular tools able to differentiate E. histolytica from E. dispar. From 2002 to 2005, 4,266 fresh stools specimens received in the setting of the National program of food handlers' control were analysed by optical microscopy. Twelve (2.8 per thousand) were positive for the presence of four nuclei cysts identified as E. histolytica/E. dispar. Extraction of DNA from the 12 samples, followed by specific amplifications of E. histolytica and E. dispar SSU rDNA, showed that 11 samples (92%) were positive for E. dispar and negative for E. histolytica. Sequencing analysis of 8 PCR products permitted to verify the results obtained with conventional PCR. The remaining sample was negative by PCR amplifying E. histolytica DNA or E. dispar DNA specifically, although it did not show any inhibition. It probably contains protozoan cysts genetically distinct from these two species but morphological similar. Estimation of relative proportions between E. histolytica and E. dispar in cyst carriers showed that all explored individuals harboured the non pathogenic E. dispar strains. This result highlights the need of use in this population of complementary tests that allow specific diagnosis and obviate unnecessary chemotherapy. PMID- 18416249 TI - Evaluation of circulating cathodic antigen (CCA) strip for diagnosis of urinary schistosomiasis in Hassoba school children, Afar, Ethiopia. AB - A total of 206 urine samples collected from Hassoba Elementary schoolchildren, Afar, Ethiopia, a low Schistosoma haematobium endemic setting, was diagnosed to evaluate the performance of CCA strip using double references, urine filtration technique and urinalysis dipstick (Combur 1.0 Test) that detect schistosome eggs and blood in urine, respectively. The former was used as a gold standard reference method. Sensitivity, specificity, positive and negative predictive values for the CCA were 52%, 63.8%, 56.7% and 59% respectively, with reference to urine filtration technique whereas these parameters were 50.4%, 62.4%, 55.6% and 57.5% respectively, with reference to Combur 10 Test. 47 S. haematobium egg positive children were found negative by CCA strip while 38 egg-negative children were found positive by CCA strip. Moreover, among the pre-tests done in duplicate, inconsistent results were also recorded. Assays were also compared with regard to the cost of equipment and reagents, speed and simplicity of use. Though CCA strip was found to be rapid and could be performed with minimal training, it was found to be expensive (US $ 4.95 per test) to use it for large scale field use even if its diagnostic value would have been satisfactory. Further development and standardization of the CCA strip are required for its applicability for field use. It is also recommended that its cost per strip should be substantially cut down if it is to be used in poor schistosomiasis endemic countries. PMID- 18416250 TI - Infection of foxes by Echinococcocus multilocularis in urban and suburban areas of Nancy, France: influence of feeding habits and environment. AB - This study evaluated the impact of biological and environmental factors on the infection of red foxes (Vulpes vulpes) by Echinococcus multilocularis in an endemic area of north-east France. From January 2004 to April 2006, 127 foxes were examined for E. multilocularis and their stomach contents analysed. The effect of year, season, age, sex and urbanisation level on E. multilocularis presence was estimated using a General Linear Model (GLM) with logit link, (i.e. logistic regression). Urbanisation level was the only influencing factor, with a decreasing gradient from rural [54%, CI 95% (40-68)] to peri-urban [31%, CI 95% (15-52)] and urban area [4%, CI 95% (0.7-15)]. The consumption of Arvicola terrestris and Microtus sp., grassland species, the main presumed intermediate hosts of E. multilocularis, was studied by the same approach. The two species were consumed less in the urban area and more in autumn than in spring. Anthropogenic food consumption was linked to urbanisation and to age. The frequency of anthropogenic food consumption decreased in the rural area. A global model explaining the presence of E. multilocularis and including urbanisation level and diet was then elaborated. Independently of urbanisation, there was a suggestion of less E. multilocularis infection with anthropogenic food consumption. Red foxes consuming Microtus sp. and A. terrestris had higher worm burden than those that did not. The results suggest that the decreasing gradient observed from rural to urban area is linked to behaviour and feeding habits. PMID- 18416251 TI - Epidemiological and clinical patterns of trichinellosis in Bulgaria from 1995 to 2002. PMID- 18416252 TI - [Aedes aegypti oviposition in response to NPK fertilizers]. AB - Fertilizers are mineral associations intended to bring to the plants nutritive complements necessary to their growth. Modern fertilizers (NPK) combine the three basic elements which are the nitrogen (N), phosphorus (P) and potassium (K). In this study, we investigated in tunnel apparatus the influence of aqueous solutions containing low, moderate and high concentrations of NPK on the oviposition of Aedes aegypti. The results showed that the solutions containing moderate concentrations (NK = 17-33 mg/l and P = 23-47 mg/l) attracted significantly more gravid females than distilled water (P < 0.001). Conversely, the solutions containing either low or high concentrations of NPK (NK = 8 mg/l and P = 12 mg/l; NK = 50 mg/I and P = 70 mg/l) did not induce significant attraction (P > 0.05). These findings suggest that NPK fertilizers may influence the egg-laying behaviour of Ae. aegypti in field situations. PMID- 18416253 TI - The effectiveness of a pyriprole (125 mg/ml) and a metaflumizone (150 mg/ml) combined with amitraz (150 mg/ml) spot-on treatment in preventing Phlebotomus perniciosus from feeding on dogs. AB - A controlled clinical trial was performed to assess the effectiveness of a pyriprole (125 mg/ml) and a metaflumizone (150 mg/ml) combined with amitraz (150 mg/ml) spot-on treatment (recommended dosage) in preventing adult female sandflies (Phlebotomus perniciosus) from feeding on dogs. Sandfly mortality was also assessed. Twelve beagle dogs were used in the study. Prior to treatment they were checked for their attractiveness to sandflies, ranked accordingly to generate partner triplets of equivalent sensitivity to sandflies: four control dogs, four treated with the pyriprole and four with the metaflumizone spot-on. The dogs were challenged with 50 unfed adult female sandflies (8-10 days old), in cages for one hour on Day 1 and Day 7. The sandflies were checked for blood feeding and mortality at one hour, 24 hours and 48 hours after exposure to the dogs. A very poor anti-feeding effect (near 7%) was seen on sandflies with the metaflumizone combined with amitraz and no antifeeding effect was seen with pyriprole. The sandfly mortality effect as a result of exposure to treated dogs was under 20% for the two spot-on. The two formulations could not be proposed in a leishmaniosis prevention program. PMID- 18416254 TI - Multifocal intraocular lenses: overview of their capabilities, limitations, and clinical benefits. PMID- 18416255 TI - Optical performance measurement and night driving simulation of ReSTOR, ReZoom, and Tecnis multifocal intraocular lenses in a model eye. AB - PURPOSE: To measure the optical properties of multifocal intraocular lenses (IOLs) for different pupil sizes. METHODS: An artificial eye was fabricated that had both the average spherical aberration and chromatic aberration levels found in the human eye. This model eye contained a saline-filled wet cell into which various IOLs can be mounted. Pupil sizes of 3 and 6 mm were explored with the system. Using the model eye, the following IOLs were examined: Alcon ReSTOR apodized diffractive, AMO ReZoom zonal refractive, and Tecnis ZM900 full-aperture diffractive. The modulation transfer function (MTF) for the lenses was calculated. The model eye was also used as a portable device to photograph nighttime driving scenes. RESULTS: For 3-mm pupils, the apodized and full aperture diffractive IOLs balance contrast between near and distance vision, whereas the zonal refractive IOL performs poorly for near vision. For 6-mm pupils, the apodized diffractive shifts performance from near vision to distance vision, whereas the zonal refractive and full-aperture diffractive IOLs continue to balance performance between distance and near. Subjectively, the night driving photographs showed much more stray light artifacts for the zonal refractive and the full-aperture and apodized diffractive IOLs. CONCLUSIONS: Under dark conditions, the shift of optical performance of the apodized diffractive lens towards distance vision reduces artifacts that appear under night driving conditions. These artifacts remain for the zonal refractive and full-aperture diffractive lenses. PMID- 18416256 TI - Visualization of the retinal image in an eye model with spherical and aspheric, diffractive, and refractive multifocal intraocular lenses. AB - PURPOSE: To present a method that visually demonstrates how spherical, aspheric, diffractive, and refractive multifocal intraocular lenses (IOLs) process light received from the cornea. METHODS: Monochromatic green light was projected through an Average Cornea Eye (ACE) Model with a cornea in front of the IOL. The model simulates a human cornea with average spherical aberration and visualizes the converging bundle of light leaving the IOL. Additionally, a US Air Force target was projected through the model, and the projected (retinal) image was captured. Various IOLs of differing designs were evaluated using this test setup. Multifocal IOLs included the aspheric diffractive Tecnis ZM900 and ZMA00 lenses; the refractive ReZoom NXG1 lens; the spherical AcrySof ReSTOR SA60D3 apodized diffractive lens; and the spherical diffractive CeeOn 811E lens. Monofocal IOLs included the spherical CeeOnEdge 911A IOL and the aspheric SofPort LI61AO, AcrySof IQ SN60WF, and Tecnis Z9000 and ZA9003 IOLs. RESULTS: The light paths of the different diffractive and refractive multifocal IOLs showed the variations in the processing of incoming light, illustrating the functional differences of IOL concepts. The US Air Force target projections in the ACE Model gave an impression of the functional optical quality of the different lenses. The value of this visualization method was demonstrated by comparing the results with modulation transfer function measurements. CONCLUSIONS: This visualization technique furthers the understanding of the working principles and quality of the retinal images produced by different mono- and multifocal IOLs. PMID- 18416257 TI - Long-term experience with mixing and matching refractive array and diffractive CeeOn multifocal intraocular lenses. AB - PURPOSE: To compare visual performance of the refractive Array SA40N and the diffractive CeeOn 811E multifocal intraocular lenses (IOLs) and to evaluate the potential benefits of combining both multifocal IOLs in the same patient. METHODS: Two groups of cataract patients were unilaterally implanted with either the CeeOn diffractive (n = 10) or the Array refractive multifocal IOL (n = 10). Another group was bilaterally implanted with one of each multifocal IOLs (mix & match group, n = 10). Visual acuity, spectacle independence, depth of focus, contrast sensitivity, presence of photic phenomena, and patient satisfaction were assessed postoperatively. RESULTS: All eyes achieved good distance visual acuity but better uncorrected near vision was achieved with the CeeOn diffractive design. Contrast sensitivity with either multifocal IOL was at the lower limit of the normal range but when multifocal IOLs were combined in the same patient, contrast sensitivity was not significantly different from phakic controls. Defocus curves revealed a superiority of CeeOn diffractive design for near and Array refractive design for intermediate but mix & match patients performed better overall than the other patients, particularly for intermediate distances, which was reflected by total independence from spectacles in 90% of patients compared to 60% in the other groups. Visual outcomes remained unchanged over time (1 month vs 6 month vs > 3 years). CONCLUSIONS: Bilateral implantation with a diffractive multifocal IOL in one eye and a refractive multifocal IOL in the fellow eye is safe and could provide patients with better intermediate vision, increased depth of focus and contrast sensitivity, and also less dependence on spectacles. PMID- 18416258 TI - Refractive lens exchange with the diffractive multifocal Tecnis ZM900 intraocular lens. AB - PURPOSE: To assess visual outcomes and patient satisfaction after refractive lens exchange followed by bilateral implantation of the multifocal Tecnis intraocular lens (IOL). METHODS: This prospective case series involved 59 eyes of 30 patients aged 56 +/- 8 years. Fifty-seven eyes were hyperopic (+3.52 +/- 1.80 diopters [D]) and 2 eyes were myopic (-1.12 +/- 0.53 D). Near, intermediate, and distance visual acuities were assessed at 1 and 6 months postoperatively. At last follow up, patients were asked about their overall satisfaction, the occurrence of photic phenomena, difficulties driving at night, and spectacle independence. RESULTS: Six months after surgery and laser retreatment in 15 eyes, 90% of eyes achieved monocular uncorrected distance visual acuity of 20/30 or better (0.087 +/- 0.085 logMAR) and 100% of eyes could read J2 or better without correction, including 90% of eyes achieving J1 or better (0.133 +/- 0.095 logMAR). Evaluation of visual performance at 1 month versus 6 months (n = 44 eyes, no retreatment) revealed a considerable improvement of the uncorrected (0.175 +/- 0.122 vs 0.127 +/- 0.094 logMAR; P = .005) and distance-corrected (0.099 +/- 0.057 vs 0.068 +/- 0.031 logMAR; P = .001) near visual acuity whereas mean refractive errors and distance visual acuity remained unchanged. Overall, 96.4% of patients were very satisfied with the procedure and would choose the same lens again. The majority of patients (92.8%) were totally free from spectacles with only 7.2% occasionally wearing glasses for intermediate tasks. CONCLUSIONS: The multifocal Tecnis ZM900 IOL provides good distance and near vision after refractive lens exchange and a period of neuroadaptation. However, laser vision correction might be required to achieve emmetropia as well as spectacles to achieve good intermediate vision. PMID- 18416259 TI - Intermediate vision and reading speed with array, Tecnis, and ReSTOR intraocular lenses. AB - PURPOSE: To retrospectively evaluate reading performance at intermediate distances of three types of multifocal intraocular lenses (IOLs) under different light conditions based on reading acuity and reading speed tests. METHODS: Thirty cataract patients (60 eyes) randomly assigned to receive the Array SA40N lens (n = 20 eyes), the Tecnis ZM001 lens (n = 20 eyes), and the ReSTOR lens (n = 20 eyes) were retrospectively tested for their reading abilities at distances of 40 cm, 60 cm, and 80 cm under low light conditions (6 cd/m2) and bright light conditions (100 cd/m2). RESULTS: LogMAR distance-corrected acuity in dim (bright) light at 40 cm was 0.456 (0.363) (Array), 0.446 (0.293) (ReSTOR), and 0.299 (0.140) (Tecnis); at 60 cm, it was 0.417 (0.259) (Array), 0.448 (0.318) (ReSTOR), and 0.381 (0.121) (Tecnis); at 80 cm, it was 0.359 (0.195) (Array), 0.395 (0.245) (ReSTOR), and 0.373 (0.124) (Tecnis). Uncorrected reading speed (words/min) in dim (bright) light at 40 cm was 25 (69) (Array), 58 (138) (ReSTOR), and 110 (166) (Tecnis); at 60 cm, it was 65 (129) (Array), 58 (129) (ReSTOR), and 135 (173) (Tecnis); and at 80 cm, it was 59 (153) (Array), 79 (164) (ReSTOR), and 121 (176 (Tecnis). CONCLUSIONS: Our study indicates that Array, ReSTOR, and Tecnis IOLs provide functional intermediate vision in bright light. In dim light, however, the Tecnis IOL provides better uncorrected reading speed at intermediate distances than Array and ReSTOR IOLs. PMID- 18416260 TI - Visual function with bilateral implantation of monofocal and multifocal intraocular lenses: a prospective, randomized, controlled clinical trial. AB - PURPOSE: To evaluate visual function of three types of multifocal intraocular lenses (IOLs) and one monofocal IOL (as the control group) after cataract surgery. METHODS: One hundred fourteen patients participated in a prospective, randomized, controlled clinical study and received monofocal Tecnis Z9000 (AMO) (n = 24, 48 eyes); symmetric diffractive multifocal Tecnis ZM900 (AMO) (n = 26, 52 eyes); zonal refractive multifocal ReZoom (AMO) (n = 32, 64 eyes); and asymmetric diffractive multifocal TwinSet (Acri.Tec) (n = 32, 64 eyes) IOLs. RESULTS: Mean binocular distance best spectacle-corrected visual acuity (BSCVA) (logMAR) was 0.05 for controls, 0.08 for ZM900, 0.07 for ReZoom, and 0.11 for TwinSet, with mean binocular distance BSCVA at near of 0.49, 0.06, 0.22, and 0.11, respectively. Mean contrast sensitivity was better for the monofocal IOL group than for the multifocal IOLs. Patients assigned to TwinSet had less favorable contrast sensitivity scores. Patients with monofocal IOLs had more frequently recommended near addition (74%) than those with multifocal IOLs. Patients with refractive ReZoom had also recommended near addition more frequently than the two diffractive groups. The percentage of dysphotopsia phenomena was 81% in patients with diffractive multifocal ZM900 compared with 48% in patients with monofocal IOLs, 53% with refractive ReZoom, and 47% with diffractive TwinSet. CONCLUSIONS: The monofocal IOL showed better visual function and lesser photic phenomena than multifocal IOLs but patients were spectacle dependent. ReZoom provided better distance BSCVA than the TwinSet diffractive model. Patients with Tecnis and TwinSet diffractive multifocal IOLs were more spectacle independent than patients with ReZoom. Patients with TwinSet had the worst visual function. Patients implanted with the Tecnis diffractive ZM900 were those reporting more photic phenomena. PMID- 18416261 TI - Visual function and change in quality of life after bilateral refractive lens exchange with the ReSTOR multifocal intraocular lens. AB - PURPOSE: To evaluate vision and quality of life and explore the correlation between vision and change of quality of life after bilateral refractive lens exchange of the Acrysof SA60D3 ReSTOR multifocal intraocular lens (IOL) for correction of ametropia in presbyopic populations. METHODS: Pre- and postoperatively, the National Eye Institute Refractive Error Quality of Life Instrument (NEI-RQL) was self-administered by 30 consecutive refractive lens exchange patients, including 19 myopes and 11 hyperopes. The change of NEI-RQL scores was determined for each subscale covering a specific aspect of quality of life. Refraction, visual acuity, and contrast sensitivity function were evaluated 3 and 6 months postoperatively. RESULTS: Postoperatively, all patients achieved binocular uncorrected distance and near visual acuity of 20/30 or better. Photopic contrast sensitivity function decreased significantly at high spatial frequencies (P < .05). Overall scores of expectations, activity limitations, dependence on correction, appearance, and satisfaction were significantly higher postoperatively (P < .05). Greater improvement in near vision and dependence on correction was achieved in hyperopes. However, glare scores were significantly lower postoperatively (P < .001). Lower preoperative uncorrected near visual acuity was associated with higher scores for dependence on correction in myopes (r = 0.51, P = .027), and overall patient satisfaction correlated significantly with postoperative uncorrected distance visual acuity (r = -0.44, P = .015). CONCLUSIONS: Refractive lens exchange with the ReSTOR IOL in presbyopic patients provided good visual function and better patient satisfaction compared with preoperative corrections. Improvement in vision-related quality of life was most evident in hyperopes. A slight reduction in contrast sensitivity function appears to have no influence on quality of life. PMID- 18416262 TI - Wavefront-guided excimer laser vision correction after multifocal IOL implantation. AB - PURPOSE: To investigate the use of wavefront-guided LASIK after multifocal intraocular lens (IOL) implantation to correct residual ametropia and higher order aberrations. METHODS: In a prospective, nonrandomized case series, wavefront-guided LASIK was performed in 27 eyes (19 patients) after multifocal IOL implantation (Tecnis diffractive IOL, n = 20; ReSTOR diffractive IOL, n = 4; ReZoom refractive IOL, n = 3) using the VISX STAR S4 IR excimer laser. Visual acuity, manifest refraction, and wavefront error were examined pre- and 3 months postoperatively. RESULTS: In the Tecnis group, results before (after) LASIK were: sphere +1.06 +/- 0.77 diopters (D) (-0.03 +/- 0.28 D; P = .0001), cylinder -1.13 +/- 0.73 D (-0.14 +/- 0.25 D; P = .00004), distance uncorrected visual acuity (UCVA) 20/45 +/- 0.09 (20/29 +/- 0.16; P = .00004), near UCVA 20/30 +/- 0.24 (20/25 +/- 0.16; P = .001), and higher order aberrations (4-mm pupil) 0.14 +/- 0.05 microm (0.18 +/-0.03 microm; P = .02). Distance and near best spectacle corrected visual acuity (BSCVA) did not change. In the ReSTOR group, results before (after) LASIK were: sphere +0.75 +/- 0.56 D (+0.13 +/- 0.22 D), cylinder 1.50 +/- 0.47 D (-0.13 +/- 0.22 D), distance UCVA 20/40 +/- 0.07 (20/26 +/- 0.07), near UCVA 20/44 +/- 0.05 (20/25 +/- 0.0), and higher order aberrations (4 mm pupil) 0.14 +/- 0.03 microm (0.20 +/- 0.02 microm). Distance and near BSCVA did not change. In the ReZoom group, results before (after) LASIK were: sphere +0.08 +/- 1.20 D (0.00 D), cylinder -0.83 +/- 0.120 D (0.00 D), distance UCVA 20/40 +/- 0 (20/25 +/- 0), near UCVA 20/60 +/- 0.09 (20/150 +/- 0.18), and higher order aberrations (4-mm pupil) 0.43 +/- 0.04 microm (0.39 +/- 0.03 microm). Patients lost one line of distance BSCVA and two lines of near BSCVA. CONCLUSIONS: Wavefront-guided LASIK is safe and effective in diffractive multifocal IOLs to correct residual refractive error, but higher order aberrations did not improve. It is not recommended in refractive multifocal IOLs, as these cannot be measured reliably with current wavefront sensors. PMID- 18416263 TI - Problems in the measurement of wavefront aberration for eyes implanted with diffractive bifocal and multifocal intraocular lenses. AB - PURPOSE: To determine the origin of the wavefront aberration as measured in eyes fitted with diffractive intraocular lenses (IOLs). METHODS: Wavefront aberration was measured by Shack-Hartmann aberrometry in 2 eyes fitted with diffractive bifocal contact lenses and 10 eyes implanted with Acrysof ReSTOR multifocal IOLs. RESULTS: Problems were encountered as a result of the multiple wavefronts generated by the diffractive lenses and their dependence on wavelength. These led to ambiguities in the positions of the Shack-Hartmann spot images and in the form of the derived wavefronts. CONCLUSIONS: Aberrometry may not provide reliable information on the wavefront aberration associated with either the distance or near power of diffractive IOLs. PMID- 18416264 TI - Intraindividual comparison of the effect of training on visual performance with ReSTOR and Tecnis diffractive multifocal IOLs. AB - PURPOSE: Several studies have shown that after multifocal intraocular lens (IOL) implantation visual function improves gradually over a period of several months. To accelerate this learning process, a prospective, intraindividual comparative study was performed to investigate the efficacy of a special visual training program on the postoperative visual performance. METHODS: Sixteen patients with bilateral phacoemulsification and multifocal IOL implantation in both eyes (Alcon ReSTOR [n = 8] and AMO Tecnis ZM900 [n = 81) received computer-based visual training 6 weeks postoperatively based on the concept of perceptual learning of discrimination line orientations. The training was performed over 2 weeks in six sessions in one eye of each patient. The untrained fellow eye served as control. Before and after the training period and at 6 months, orientation visual acuity, distance visual acuity, contrast sensitivity, and near visual acuity for different contrast levels were assessed. RESULTS: After 2 weeks of training, the mean improvement of orientation visual acuity in the trained eyes was 82%, which was significantly higher than the control eyes (P < .001). Contrast sensitivity and near vision under different contrast levels showed a significant benefit of training. The superior function of the trained eyes was still present at 6 months. CONCLUSIONS: Visual performance after multifocal IOL implantation can be significantly accelerated by a specific 2-week training program. This effect is sustained over a 6-month period. PMID- 18416265 TI - Comparison of Acri. Smart multifocal IOL, crystalens AT-45 accommodative IOL, and Technovision presbyLASIK for correcting presbyopia. AB - PURPOSE: To compare the visual outcomes of two intraocular lenses (IOLs) (Acri. Smart [multifocal] and AT-45 [accommodative]) for correction of distance and near visual acuity with a LASIK procedure for presbyopia (presbyLASIK) and a control group fitted with a standard distance-correcting monofocal IOL (Acrysof). METHODS: Distance and near decimal visual acuities were measured under similar conditions before and 6 months postoperatively in 45 patients (87 eyes) who comprised four groups: Acri. Smart IOL, AT-45 IOL, presbyLASIK, and control (monofocal IOL). All acuity measures were standardized at 20 m (distance, projected chart) and 40 cm (near). All visual acuity measures were taken in the same ambient light conditions and contrast. All data were collected in a randomized, masked style. RESULTS: Pre- and postoperative distance mean visual acuities for the Acri. Smart IOL, AT-45 IOL, presbyLASIK, and controls groups, respectively, were 0.15 +/- 0.21 and 0.66 +/- 0.35; 0.31 +/- 0.14 and 0.68 +/- 0.28; 0.35 +/- 0.15 and 0.80 +/- 0.33; and 0.32 +/- 0.18 and 0.59 +/- 0.29. Pre- and postoperative near mean visual acuities for the Acri. Smart IOL, AT-45 IOL, presbyLASIK, and control groups, respectively, were 0.13 +/- 0.21 and 0.62 +/- 0.35; 0.43 +/- 0.22 and 0.51 +/- 0.27; 0.15 +/- 0.20 and 0.68 +/- 0.33; and 0.40 +/- 0.29 and 0.30 +/- 0.13. Distance and near visual acuities improved significantly after surgery (P < .05) except near acuities for the AT-45 IOL and control groups. CONCLUSIONS: For these patients, distance and near acuities improved with presbyLASIK and Acri. Smart IOLs when compared with controls. In those patients treated with the AT-45 IOL, average distance visual acuity improved but not near visual acuity. PMID- 18416266 TI - Visual results following implantation of a refractive multifocal IOL in one eye and a diffractive multifocal IOL in the contralateral eye. AB - PURPOSE: To assess binocular visual results in patients who were scheduled to undergo cataract/refractive lens exchange with multifocal IOL implantation using a "mix & match" approach. METHODS: This prospective study enrolled 40 eyes of 20 patients aged between 44 and 78 years (median age = 58.1 years) implanted with a refractive multifocal IOL (ReZoom) in their dominant eye and a diffractive multifocal IOL (Tecnis) in their non-dominant eye. Near, intermediate, and distance vision were assessed at 2 months following implantation. Patients underwent refractive lens exchange or cataract surgery in the dominant eye first, followed by the non-dominant eye 1 to 2 weeks later. Emmetropia was the goal for all surgeries. RESULTS: Preoperatively, the mean spherical equivalent refraction was +2.019 +/- 1.417 diopters (D) (median: +2.063 D). The mean binocular distance decimal visual acuity was 1.06 +/- 0.60 D, the mean binocular intermediate decimal visual acuity was 0.50 +/- 0.90 D, and the mean binocular near decimal visual acuity was 1.10 +/- 0.40 D. CONCLUSIONS: Preliminary visual outcomes in this series of patients indicate good results at all distances. PMID- 18416267 TI - Postoperative changes in refraction and anterior chamber depth in different multifocal intraocular lenses. AB - PURPOSE: To investigate the axial shift of multifocal intraocular lenses (IOLs) during the first months after implantation and to determine its potential effect on uncorrected distance visual acuity. METHODS: This study comprised 8 patients (16 eyes) who received the Tecnis multifocal IOL and 8 patients (16 eyes) who received the ReSTOR multifocal IOL. Anterior chamber depth was measured with a Scheimpflug camera, and visual acuity and subjective manifest refraction were determined at 1 day, 1 week, and 1 and 3 months after surgery. RESULTS: The spherical equivalent refraction in the ReSTOR and Tecnis eyes, respectively, was +0.125 +/- 0.412 diopters (D) and +0.306 +/- 0.481 D at day 1, -0.375 +/- 0.280 D and -0.188 +/- 0.438 D at 1 week, -0.475 +/- 0.337 D and -0.018 +/- 0.512 D at 1 month, and 0.297 +/- 0.267 D and -0.036 +/- 0.393 D at 3 months. Anterior chamber depth in the ReSTOR and Tecnis eyes, respectively, was 3.18 +/- 0.1 mm and 3.16 +/- 0.17 mm at day 1, 3.18 +/- 0.10 mm and 3.17 +/- 0.20 mm at 1 week, 3.29 +/- 0.22 mm and 3.18 +/- 0.25 mm at 1 month, and 3.14 +/- 0.19 mm and 3.20 +/- 0.14 mm at 3 months. The change in axial position did not correlate with the change in refraction. CONCLUSIONS: Multifocal IOLs change their axial position slightly during the first postoperative months. The ReSTOR IOL showed a myopic change of approximately -0.50 D during this time period, which did not correlate with the change in axial position. PMID- 18416268 TI - Wavefront measurements of diffractive and refractive multifocal intraocular lenses in an artificial eye. AB - PURPOSE: To investigate whether wavefront measurements taken in eyes that have implanted multifocal intraocular lenses (IOLs) are reliable when creating laser refractive surgical treatments to remove residual refractive error from these eyes. METHODS: A specially designed fluid-filled model eye holding a multifocal IOL was refracted using a commercial wavefront eye refractor. The wavefront findings were then compared to the expected refractive error. The fluid-filled model was designed to match the optical characteristics of a human eye and to hold the IOL at the same distance from the cornea as in a human eye. Lenses tested were AMO Tecnis ZM900 series (diffractive design, +25.00 D, +4.00 D add), AMO ReZoom NXG1 series (refractive design, 25.00 D, +4.00 D add), and AMO 811E (diffractive with aspheric design 25.00 D, +4.00 add). The visible pupil diameter was 5.75 mm. RESULTS: Diffractive bifocal IOLs were found to create spot doubling in the Shack-Hartmann sensor image as expected from theoretical considerations. However, the residual sphere power and cylinder power reported were those expected. In addition, the higher order aberrations reported were reasonable in light of expected results. Refractive multifocal IOLs were found to create distorted Shack-Hartmann images as expected. The residual sphere power and cylinder power reported were not those expected nor were the higher order aberrations. In addition, the higher order values were sensitive to the position of the eye with respect to the refractor. CONCLUSIONS: Wavefront measurements from eyes with diffractive IOLs can be used, with caution, to plan laser refractive treatments. Wavefront measurements from eyes with refractive multifocal IOLs should not be used to plan post-implant laser refractive correction. PMID- 18416270 TI - How are drugs approved? Part 3. The stages of drug development. AB - The first article of this series on the drug development process described the historical evolution of the U.S. Food and Drug Administration (FDA), and last month's article reviewed the ethical foundations of clinical research. Before a new drug is marketed, a sequence of preclinical investigations and three phases of clinical studies are conducted. This drug development process involves the FDA, pharmaceutical companies (sponsors), clinical investigators, and institutional review boards. This article further describes this aspect of the drug development process. PMID- 18416271 TI - Music as a healing art for older adults. AB - There is increasing evidence of the importance of regular mental and physical exercise to maximize overall health and functioning in older adults. However, many individuals find that reduced strength or disabilities prevent them from participating in the kinds of exercise they enjoyed when they were younger. Music can provide the important benefits of both mental and physical stimulation to even frail older adults. Whether using Conductorcise for aerobic exercise, enjoying the communal experience of singing in a choir, or quietly reflecting on a music recording, music can serve as a healing art for older adults. PMID- 18416272 TI - Domestic violence: a worldwide exploration. AB - The concept of domestic violence is important to nursing because further knowledge of this phenomenon can help drive more effective intervention and prevention approaches. Domestic violence, also known as gender violence, interpersonal violence, and domestic abuse, is a daily and often a deadly fact of life for millions of women and girls around the world. Attacks are perpetrated in settings that include public streets, at places of employment, in the home, while serving in the military, and in prisons and jails. The costs of this violence, both monetary and in human lives, are tremendous. Although nurses are in a unique position to screen for domestic violence and provide assistance and advocacy for victims, barriers exist related to victim disclosure and staff training and comfort levels. Advanced practice nurses can address the issue directly through appropriate treatment facilitation, including both physical and psychosocial intervention. PMID- 18416273 TI - Behavioral activation and rehabilitation. AB - Rehabilitation from a mental or physical disorder can become complicated when patients feel overwhelmed by symptoms and unable to make progress. Newer behavioral approaches have been developed that can help these patients. This article describes behavioral activation techniques designed to address depression. PMID- 18416274 TI - Adolescents' attitudes toward schizophrenia, depression and PTSD. AB - The objective of this study was to compare adolescents' attitudes toward schizophrenia, depression, and posttraumatic stress disorder (PTSD). Stigmatizing attitudes toward these three mental disorders were evaluated in 325 senior students from medical, commercial, and grammar high schools in Croatia using a 45 item questionnaire. Results indicated that stigmatizing attitudes toward PTSD were at the same level as attitudes toward depression and were less stigmatizing than were attitudes toward schizophrenia. Negative attitudes were the lowest among medical students. Gender differences existed only for attitudes toward depression. Stigmatizing attitudes toward PTSD were not as high as expected. All of the students believed mental disorders are different from other disorders. Specific knowledge of psychiatry has been shown to reduce stigma. The nonaddictive nature of psychotropic medications should receive greater emphasis when teaching psychiatry. PMID- 18416276 TI - Practical legal issues in end of life care. PMID- 18416277 TI - Is nursing education up to the job? PMID- 18416278 TI - Therapeutic skills in nursing: have they been lost? PMID- 18416279 TI - The role of the cancer nurse coordinator in neurology. PMID- 18416280 TI - Increasing capacity in general practice. PMID- 18416281 TI - Levetiracetam and bleeding disorders. AB - Six studies were conducted in healthy male volunteers to evaluate the effect of levetiracetam on bleeding time. In three open-label studies, a single dose of levetiracetam (250, 500, or 1000 mg, respectively) was administered 12 hours after acetylsalicylic acid (aspirin). Bleeding time increased by 3.5% to 30% relative to baseline, but the effect was not dose-related and not clinically relevant. In a fourth open-label study, levetiracetam was administered twice daily for 4 days, with aspirin administered with the penultimate dose of levetiracetam. The other two studies had a double-blind, placebo-controlled crossover design: levetiracetam or placebo was administered twice daily for 4.5 days and then aspirin was coadministered with the final dose. In the open-label multiple-dose study, the bleeding time increase was more pronounced after repeated levetiracetam doses and ingestion of aspirin than after a single 250-mg dose. However; no clinically relevant change in bleeding time or difference from placebo was observed in the double-blind, crossover studies. Except for two subjects in a crossover study, one of whom received placebo, no absolute bleeding time value was above the normal range of 4-8 minutes. These results indicate that levetiracetam does not produce clinically significant increases in bleeding time in healthy male volunteers. Further a review of clinical trials suggests that levetiracetam does not appear to cause clinically significant or relevant hematological adverse events suggestive of underlying hematological disorders. PMID- 18416282 TI - Obstacles to the use of intravenous tissue plasminogen activator for acute ischemic stroke. Is time the only barrier? AB - BACKGROUND AND AIMS: The short time window is frequently cited as the main reason for exclusion of intravenous tissue plasminogen activator (tPA) in acute stroke. Identifying and circumventing barriers to thrombolysis other than time could increase the frequency of treatment. The goal of this study was to identify whether the rate of treatment with tPA would increase if time window was not an obstacle to treatment. METHODS: In four hospitals we prospectively recorded the rate of tPA use in consecutive patients admitted with acute ischemic stroke and in those admitted within 3 hours, the reasons why thrombolysis was not given, and the potential gain in the rate of tPA use if all patients had been admitted within 3 hours considering all exclusion criteria other than time. RESULTS: We recruited 486 patients (258 men; mean age, 70.4 +/- 13.5 years), of whom 154 (31.7%) were admitted within 3 hours. The time of stroke onset was unknown in 28 (5.8%). The rate of tPA use was 11.1% in the whole study population and 35.1% in those admitted within 3 hours. In 'eligible' patients, TIA in 33 patients (21.4%), age >80 years in 27 (17.5%), mild stroke in 27 (17.5%), rapidly recovering stroke in 9 (5.8%), a too long door-to-needle time in 8 (5.2%), and severe hypertension in 5 (3.2%) were the main reasons for not to treat. Two patients were not treated despite the absence of documented reasons. If all patients had hypothetically been admitted within 3 hours, the rate of tPA use would have increased from 11.1% up to 27.9% of the whole population and up to 37.4% if aged patients and those with no documented reasons or with a rapidly recovering stroke had also been considered for treatment. CONCLUSIONS: Delayed presentation to hospital is the main factor limiting the use of thrombolysis in acute stroke, but additional patients would be treated if those with no contraindication, advanced age or rapidly recovering stroke were considered as eligible. PMID- 18416283 TI - Autonomic function in tension-type headache. AB - The pathogenesis of tension-type headache (TTH) is poorly understood. TTH is mostly associated with the sympathetic nervous system. Reports considering autonomic nervous system functioning with regard to TTH are limited. Autonomic nervous system disorders could however play a role in the etiopathology of TTH. Compared with sympathetic nervous system functioning, techniques for evaluating parasympathetic nervous system activity are more limited. Therefore, the aim of the study was to measure heart rate recovery (HRR), an index of vagal activity, in TTH patients. Thirty-seven patients with TTH (15 with episodic and 22 with chronic TTH) and 37 control subjects underwent exercise tolerance testing according to modified Bruce protocol. Heart rate recovery (HRR) at 1 and 3 minutes (HRR1 and HRR3) were calculated. HRR 1 and 3 were found to be similar in patients and controls. However the resting heart rate in patients with episodic TTH was found to be significantly lower than the resting heart rate in patients with chronic TTH and control subjects. These results suggest that parasympathetic nervous system function is not affected in TTH patients. However sympathetic nervous system tonus, which is evaluated by resting heart rate, is low in patients with episodic TTH, suggesting sympathetic hypofunction. PMID- 18416284 TI - Recurrent miller fisher syndrome with abnormal terminal axon dysfunction: a case report. AB - Miller Fisher syndrome (MFS) is a localized variant of Guillain-Barre syndrome (GBS), characterized by ophthalmoplegia, areflexia, and ataxia. Recent neurophysiological studies have suggested that abnormal terminal axon dysfunction occurs in some cases of Miller Fisher syndrome and Guillain-Barrd syndrome. We present a rare case report of recurrent MFS with abnormal terminal axon dysfunction. To the best of our knowledge, this is the first case report of recurrent MFS with terminal axon dysfunction that persisted up to nine months after the initial presentation of the second attack with positive antiganglioside antibodies and full clinical recovery. PMID- 18416285 TI - Left recurrent laryngeal palsy after left total hip arthroplasty; stretch injury due to inappropriate positioning in the lateral approach? AB - An 80-year-old man is described with a left-sided recurrent laryngeal nerve palsy after primary total hip arthroplasty. Intubation was uncomplicated. Aetiology is discussed. It might be caused by intubation injury, stretch-induced injury due to cervical hyperextension or inappropriate positioning during surgery in the lateral approach or a combination. PMID- 18416286 TI - Acquired progressive spastic paraparesis due to neurobrucellosis: a case report. AB - A 39-year-old man with a 4-month history of transient pins and needles sensations occurring below the waist while walking and difficulty walking presented to our outpatient clinic. He had an approximate 1-year history of bilateral hearing loss, the etiology of which was unknown. His symptoms had been progressive, and there was no significant family history. He demonstrated a spastic gait and required assistance for walking. Deep tendon reflexes were hypertonic; a sensation deficit was defined as originating from the 12th thoracic vertebra. Babinski's sign was positive bilaterally. Sphincter abnormalities were seen in the patient's bladder and bowel functions. Cerebral and spinal magnetic resonance images with contrast media were unremarkable. An analysis of the patient's cerebrospinal fluid was consistent with neurobrucellosis. Owing to spastic paraparesis and hearing loss, the diagnosis of neurobrucellosis was made. Combined antimicrobial therapy was started and continued 6 months. His neurologic condition improved, and he was able to walk without help after 3 months' treatment. Our case illustrates that acquired progressive spastic paraparesis may occur during the course of neurobrucellosis. Neurobrucellosis should be borne in mind when patients present with spastic paraparesis. PMID- 18416287 TI - A Foix's syndrome revealing "mirror" giant intracavernous aneurysms an illustrative case report of a therapeutic dilemma. PMID- 18416289 TI - A case of 'tropical' myelopathy. AB - We present a case of lower limb sensory disturbances and weakness in a patient originating from Mali. MRI showed a diffuse myelopathy of the cervical and thoracic spinal cord. Serological evaluation of blood and cerebrospinal fluid pointed towards schistosomiasis as the cause. Histological confirmation was made on bladder-biopsy. Treatment with praziquantel and steroids brought marked clinical improvement. This case illustrates the need to keep in mind more exotic causes of myelopathy in those patients coming from endemic regions. PMID- 18416288 TI - Isolated nontraumatic abducens nerve palsy. AB - INTRODUCTION: Abducens nerve palsy (ANP) is the most common isolated palsy. The injury of the abducens nerve can occur anywhere along its long course, so differential diagnosis of ANP occasionally demands thorough investigation to find the proper cause. PATIENTS: Ten patients with isolated ANP are presented. The causes included nasopharyngeal carcinoma, diabetes, cholesteatoma of the inner ear, carotid-cavernous fistula, subarachnoid bleeding, hydrocephalus, toxic ANP, multiple sclerosis, clinically isolated syndrome suggestive of multiple sclerosis, and Tolosa-Hunt syndrome. DISCUSSION: Based on the cases presented and review of the literature, we argue that every patient with isolated nontraumatic ANP requires brain MRI as an initial diagnostic tool. If this finding remains inconclusive, additional tests including angiography and CSF examination should be performed. PMID- 18416290 TI - Tumefactive lesion in multiple sclerosis. PMID- 18416291 TI - [Current views on biliary secretion and pathogenesis of gallbladder stones]. PMID- 18416292 TI - [Obesity as a risk factor of chronic kidney disease in patients undergoing primary angioplasty]. AB - The number of patients with chronic kidney disease-CKD is still growing. Overweight and obesity present also an important problem of world public health. However, there are not many data showing possible association between obesity and incresing risk of development of renal failure recently it has been demonstrated that in obese patients secondary focal segmental glomerulosclerosis and glomerular hypertrophy appear more frequently. The aim of this study was to estimate glomerular filtration rate-GFR in patients with normal serum creatinine concentration undergoing primary angioplasty according to body mass index. The study included 1413 patients udergoing primary angioplasty for acute myocardial infarction. The following parameters were assessed: age, gender, family history of cardiovascular disease, risk factors of cardiovascular disease (hypertension, diabetes mellitus, obesity etc.), previous myocardial infarction, pre-existing heart failure, treatment given, localization of infarct, coronary stenting, serum creatinine before angioplasty, cholesterol, LDL, HDL, triglycerides, glucose, blood pressure. Of a total of 1413 patients, 1337 (94.62%, 943 M, 394 F) had correct serum creatinine concentration (below 1.5 mg/dl for men, below 1.2 mg/dl for women). Glomerular filtration rate was calculated from serum creatinine levels by using the simplified Modification of Diet in Renal Disease Study formula--MDRD, Cockcroft-Gault equation and Jeliffe formula. An average value of GFR in study group was 79.94 +/- 24.51 ml/min (Cockcroft-Gault equation), 73.02 +/- 21.96 ml/min (Cockcroft-Gault adjusted to weight), 90.37 +/- 25.1 ml/min (MDRD equation) and 77.67 +/- 21.65 ml/min (Jeliffe formula). A significant lower serum creatinine levels and GFR (assessed by 3 formulas and Cockcroft-Gault using adjusted weight) were observed in women group. In the whole study group (with normal serum creatinine levels) substantial correlation was found between age and serum creatinine concentration (r = 0.13, p > 0.001), GFR (MDRD, r = -0.37, p < 0.001, Cockcroft-Gault, r = -0.62, p < 0.001, adjusted to weight r = -0.64, p < 0.001, Jeliffe r = -0.61, p < 0.001) and also between BMI and GFR (MDRD r = 0.28, p < 0.001, Cockcroft-Gault, r = 0.31, p < 0.001, adjusted to weight r = 0.08, p < 0.001, Jeliffe r = 0.341, p < 0.001), but not with serum creatinine concentration (r = 0.03, p = 0.3). In patients with normal serum creatinine levels percentage of patients with GFR below 60 ml/min ranges from 4.79% up to 30.74%. In patients with higher BMI, higher GFR may be partially caused by glomerular hyperfiltration. Overweight or obesity are significant, but potentially changeable risk factors for development of chronic renal failure. However, chronic kidney disease is one of the complications of obesity. PMID- 18416293 TI - [Evaluation of bone mineral density in women with chronic liver diseases during perimenopausal period]. AB - Osteoporosis is the most frequently occurring metabolic diseases of bones, observed especially in women after menopause. The goal of the paper was a comparison of bone mineral density (BMD) of health women with that in perimenopausal patients with chronic liver diseases. The study was performed in 47 patients with chronic liver diseases, aged: 37-56 years. Qualification criteria included chronic type B hepatitis, chronic type C hepatitis and cirrhosis of viral aetiology. The control group consisted of 15 healthy, age matched women. All the women had been examined in order to identify other risk factors of osteoporosis development. RESULTS: The incidence of decreased BMD was statistically higher in the group of patients with chronic hepatic diseases, compared to the group of healthy subjects. No bone fracture was found in any of the examined patients. CONCLUSIONS: Routine densitometric examinations should be performed in all women in perimenopausal age with chronic liver diseases. Results of our studies indicate that in patients with liver diseases, the lowest BMD values are found in the group of patients with cirrhosis or chronic type C hepatitis. These women constitute a large risk group for secondary osteoporosis development. PMID- 18416294 TI - [Left ventricular remodelling and rebuilding of the infarcted area. Morphometric evaluation of the relationships between left ventricular mass, myocyte diameter, capillary density and fibrosis]. AB - The aim of the study was to assess the structural changes in both the infarcted region and free left ventricular (LV) wall that underly post-infarct LV remodeling in the human heart. MATERIAL AND METHODS: Morphometric evaluation was performed in 31 human hearts, as follows: group I (control)--non-cardiac death (mean age: 52.4 +/- 11.2) and group II--post-Q-wave myocardial infarction (QMI) death (mean age: 58.8 +/- 9.4). The following morphological parameters were assessed: LV mass, infarct scar extent, stenosis of the infarct-related artery (IRA), myocyte diameter, myocyte nuclear diameter, myocyte nuclear density, density of the coronary network and LV fibrosis. Morphometric measurements were performed with the use of digital image analyser Leica Q500MC. RESULTS: A significant increase in the LV mass was noted in group II in comparison with the control group (296.0 +/- 81.3 g vs 150.2 +/- 18.6 g; p < 0.00001). LV hypertrophy was associated with: an increase in the myocyte diameter within the infarcted region (increase 61%) and free LV wall (increase 35%), an increase in the myocyte nuclear diameter (increase 28.1% and 11.2%, respectively), a reduction in the myocyte nuclear density (increase 52.9% and 34.4%, respectively), a reduction in the capillary density (48.6% and 4.1%, respectively) and an increase in fibrosis within the free LV wall. Significant linear relationships were observed within the free LV wall between: LV mass and myocyte diameter (r = 0.695; p < 0.001), infarct size and myocyte diameter (r = 0.451; p = 0.046), myocyte diameter and myocyte nuclear diameter (r = 0.551; p = 0.021), myocyte diameter and miocyte nuclear density (r = -0.542; p = 0.011) and miocyte diameter and capillary density (r = -0.569; p = 0.009). Such relationships were not observed within the infarcted area. Conversely, significant relationships between a reduction in the capillary density and IRA stenosis (r(s) = -0.512; p = 0.03) as well as the infarct size (r = -0.616; p = 0.006) were noted. CONCLUSIONS: 1. Left ventricular remodelling post myocardial infarction is a heterogenous process and the observed differences are mainly quantitative in character; 2. Compensatory left ventricular hypertrophy dominates in the areas not affected by necrosis. Rebuilding of the infarcted area is constrained mainly by the changes in the capillary network and depends on the IRA status. PMID- 18416295 TI - [Prevalence of cervical spine inflammatory changes in rheumatoid arthritis patients and the value of neurological examination in their diagnosis]. AB - The aim of the study was to evaluate prevalence of cervical spine inflammatory changes, especially atlantoaxial pathology, and their possible relation to subjective and objective neurological symptoms in rheumatoid arthritis patients. 100 patients (88 female and 12 male) aged 23 to 85 (61.4 +/- 12.9), with the mean disease duration of 12.5 +/- 9.5 years were included in the study. According to radiological examination (lateral and antero-posterior X-ray of the cervical spine) supplemented by MR of the cervical spine or CT of the atlanto-axial joint in suspected cases, 26% of patients had only inflammation, next 15% of patients presented with instability of the atlanto-axial joint and 9% developed basilar invagination of the dens of axis. 18% of patients presented subaxial cervical instability. Neurological examitation was performed by independent neurologist in 99 patients, only 14 presented abnormalities suggesting cervical myelopathy. Two of them showed no patology of the cervical spine. Remaining patients presented: C1/C2 inflammation in 4 cases, anterior atlanto-axial subluxation (AAS) in two cases, basilar invagination in 4 cases and instability with medullary compression on lower cervical levels only--in two cases. There were 4 cases of coexisting C1/C2 changes with medullary compression due to discopathy and (in 3 of them) instability on lower cervical levels. In 6 cases surgical stabilisation was proposed (5 patients with basilar invagination and 1 patient with AAS and myelopathy). There was statistically significant correlation between symptoms (like: paraesthesiae, intermittent problems with hearing and seeing), neurological examination and degree of radiological damage of atlanto-axial joint. The authors concluded that careful medical history and neurological examination can be useful in making decision of further radiological diagnostic procedures of the cervical spine in rheumatoid arthritis. PMID- 18416296 TI - [Mycophenolate mofetil maintenance therapy in lupus nephritis]. AB - INTRODUCTION: The aim of this study was to assess clinical efficacy and changes in immunological parameters during maintenance therapy with mycophenolate mofetil (MMF) in patients with lupus nephritis. METHODS: Patients (n = 7) with systemic lupus erythematosus (SLE) and proliferative nephropathy confirmed by renal biopsy, in whom disease remission was induced with intravenous cyclophosphamide, received MMF (2 x 500 mg daily) for 3 months. Clinical and immunological parameters as well as drug tolerance were assessed before, and after 1 and 3 months of MMF therapy. RESULTS: MMF therapy lead to a decrease in disease activity index (SLEDAI), and maintenance of previously achieved reduction of proteinuria, improvement of renal function and immunological parameters. These changes were associated with decrease in the number of activated T cells and increase in B cells. The titer of anti-dsDNA antibodies was reduced in 5 out of 7 patients. CONCLUSIONS: Mycofenolate mofetil is effective in the maintenance therapy oflupus nephritis. A 3-month MMF treatment allows for sustained beneficial immunological profile achieved in induction therapy. PMID- 18416297 TI - [Lipid and carbohydrate disturbances in men with coronary artery disease according to age]. AB - The aim of this study was to search for the factors favouring the revealing of a coronary artery disease by comparing selected lipids and carbohydrate parameters between two groups--younger and older men with coronary artery disease. Into the study were included 60 men with stable angina confirmed by coronarography (at least 6 months after CABG, PTC A or myocardial infraction, with EF > 40%) and 33 men without atherosclerotic changes in the coronary vasseles as the control group. The two groups were divided depending on age: the younger subgroup [(under 60-ty years old-group: A with CAD, n = 30) and (B without CAD, n = l8)] and older subgroup [(over 70 years old-group: C with CAD, n = 30) and (D without CAD, n = 15)]. Men of middle age were not included. Serum semples were analyzed for the concentration of lipids parameters including apolipoprotein AI and B. The oral glucose tolerance test was carried out for all patients. In control groups (B and C) in comparison to the groups with CAD (A and C) was observed significantly higher concentration of the HDL cholesterol (by abouth 28% in younger men and 16% in older men (r with CAD = 0.40) and ApoAI (respectively by about 17% and 12.5%, r = 0.47). Weak, but positive corelation with the presence of coronary heart disease was shown for fasting glucose concentration. The younger and older men with CAD (A vs C) statistically significantly differed in concentration of HDL (a.v. 9.1 +/- 6.2 vs 46.7 +/- 11,7 ng/dl) and strongly correlated with HDL ApoAI (av. 137.6 +/- 12.6 vs 146.6 +/- 21.5 ng/dl). The negative family history was more often in younger patients. PMID- 18416298 TI - [Peripartum cardiomyopathy and preeclampsia complicated with HELLP syndrome--a case report]. AB - Peripartum cardiomyopathy (PC) and preeclampsia with HELLP syndrome are serious complications of pregnancy, but the coincidence of both in one pregnancy is extremely rare. Here, we report a case of 32-year-old primipara who in 35th Hbd presented for the first time in her life symptoms of severe heart failure (HF) in NYHA class III/IV. In 37th Hbd the diagnosis of PC was established based on clinical status and echocardiographic examination, which demonstrated a dilatation of heart chambers and impaired left ventricular systolic function with decreased ejection fraction (EF) 17%. In 37th Hbd she developed symptoms of preeclampsia complicated with HELLP syndrome (hemolysis, elevated liver enzymes, low platelets) and further a DIC syndrome as well. Because the patient was in critical condition and the foetus' life was threatened the pregnancy was terminated with urgent cesarean section. Then the patient developed shock, respiratory insufficiency and increasing renal failure. Successful treatment, included administration of pressor amines, respirator, hemodialyses, multiple fresh frozen plasma and blood transfusions. The symptoms of HELLP syndrome resolved by 9th day of treatment. Although optimal treatment of HF was administered with significant clinical improvement, the normalization of left ventricle systolic function was not observed. At 2 and 13 months follow-up, EF remained low and was 34 and 36% respectively. This allows to diagnose persistent PC. Based on the case, the issues of etiopathogenesis, treatment, prognosis and the risk of recurrence of PC and HELLP syndromes in a possible pregnancy are discussed. PMID- 18416299 TI - [Broncho-pulmonary carcinoid with the cardiac arrhythmia manifestation]. AB - We are presenting the case of a 60-year-old woman (OM ) with a bronchial pulmonary carcinoid which the only, first and dominating symptom was paroxysmal atrial tachycardia, as well as ventricular group extrasystoles. After 1.5 years of treatment with amiodarone, the patient was admitted to hospital because of haemoptysis. In the imaging examinations of the thorax, a cavernous structure of 36 x 47 mm in the III segment of the left lung was discovered. The patient underwent urgent surgery after an episode of massive pulmonary haemorrhage- surgery included excision of the upper lobe of the left lung containing the tumour and mediastinal lymph nodes. On microscopic examination, an atypical carcinoid was diagnosed. After the surgery, arrhythmia ceased and did not return for a period of 3 following years. The current imaging examinations and determinations of 5-HIO uringexcretion acid do not indicate relapse of the tumour. PMID- 18416300 TI - [Subclinical hypothyroidism]. PMID- 18416301 TI - [Efficacy and safety of TNF inhibitors--results of randomized, controlled clinical trials vs long term observational studies]. PMID- 18416302 TI - [Somatostatin analogs]. PMID- 18416303 TI - [Anti-tNFalpha therapy in chronic severe asthma]. PMID- 18416304 TI - Activation of fungal silent gene clusters: a new avenue to drug discovery. AB - The ongoing exponential growth of DNA sequence data will lead to the discovery of many natural-product biosynthesis pathways by genome mining for which no actual product has been characterised. In many cases, these clusters remain silent under laboratory conditions. New technologies based on genetic engineering are available to induce silent genes. Heterologous expression of a silent gene cluster under the control of defined promoters can be applied. Alternatively, promoters of biosynthesis genes within the genome can be exchanged by defined promoters. Most promising, however, is the activation of pathway-specific regulatory genes, which was recently demonstrated. Such regulatory genes are present in many secondary metabolite gene clusters. This approach is rendered feasible by the fact that all of the genes encoding the large number of enzymes required for the synthesis of a typical secondary metabolite are clustered and that in some cases, a single regulator controls the expression of all members of a gene cluster to a certain extent. The advantage of this technique is that only a small gene needs to be handled, and that an ectopic integration is sufficient, bypassing all limitations of homologous recombination. Most conveniently, this strategy can trigger the concerted expression of all pathway genes. The vast amount of DNA sequences in the public database represents only the beginning of this new genomics era. The activation of these gene clusters by genetic engineering will lead to the discovery of many so far unknown products and therefore represents a novel avenue to drug discovery. PMID- 18416305 TI - Total synthesis studies on macrocyclic pipecolic acid natural products: FK506, the antascomicins and rapamycin. AB - This chapter derives its inspiration from the challenges presented to total synthesis chemists, by a particular group of macrocyclic pipecolic acid natural products. Although there is considerable emphasis on the completed syntheses of the main characters (FK506 (1), the antascomycins (4 and 5) and rapamycin (7)), the overall complexity of the molecular problem has stimulated a wealth of new knowledge, including the development of novel strategies and the invention of new synthetic methods. The ingenious and innovative approaches to these targets have enabled new generations of analogues, and provided material to further probe the biology of these fascinating molecules. With pharmaceutical application as an immunosuppressant, as well as potential use for the treatment of cancer and neurodegenerative diseases, this family of natural products continues to inspire new and interesting science while providing solutions to healthcare problems of the world. PMID- 18416306 TI - Application of natural product-inspired diversity-oriented synthesis to drug discovery. AB - Natural products have played a critical role in the identification of numerous medicines. Synthetic organic chemistry and combinatorial chemistry strategies such as diversity-oriented synthesis (DOS) have enabled the synthesis of natural product-like compounds. The combination of these approaches has both improved the desired biological properties of natural products as well as the identification of novel compounds. Diversity concepts and strategies to access novel compounds inspired by natural products will be reviewed. PMID- 18416307 TI - Cheminformatics analysis of natural products: lessons from nature inspiring the design of new drugs. AB - Natural products (NPs) have evolved over a very long natural selection process to form optimal interactions with biological macromolecules. NPs are therefore an extremely useful source of inspiration for the design of new drugs. In the present study we report the results of a cheminformatics analysis of more than 130,000 NP structures. The physicochemical properties of NPs and their typical structural features are compared to those of bioactive molecules and average organic molecules. The relationship between the structure of NPs and the type of organism from which they have come has also been analyzed. The aim of this study was to identify those properties and structural features which are typical for NPs and discriminate this class of molecules from common synthetic molecules, with the ultimate goal being to provide a guide for the design of novel NP-like bioactive structures. Hopefully the results of this analysis help to eliminate the old myth about NPs as being 'too complex' or having 'bad properties', as well as help us to focus on these areas of NP structural space which are essential for biological activity, taking advantage of the process of natural selection over billions of years to guide us to new and as yet unexplored areas of the Chemical Structure Universe. PMID- 18416308 TI - Chemical-genetic approaches for exploring the mode of action of natural products. AB - Determining the mode of action of bioactive compounds, including natural products, is a central problem in chemical biology. Because many genes are conserved from the yeast Saccharomyces cerevisiae to humans and a number of powerful genomics tools and methodologies have been developed for this model system, yeast is making a major contribution to the field of chemical genetics. The set of barcoded yeast deletion mutants, including the set of approximately 5000 viable haploid and homozygous diploid deletion mutants and the complete set of approximately 6000 heterozygous deletion mutants, containing the set of approximately 1000 essential genes, are proving highly informative for identifying chemical-genetic interactions and deciphering compound mode of action. Gene deletions that render cells hypersensitive to a specific drug identify pathways that buffer the cell against the toxic effects of the drug and thereby provide clues about both gene and compound function. Moreover, compounds that show similar chemical-genetic profiles often perturb similar target pathways. Gene dosage can be exploited to discover connections between compounds and their targets. For example, haploinsufficiency profiling of an antifungal compound, in which the set of approximately 6000 heterozygous diploid deletion mutants are scored for hypersensitivity to a compound, may identify the target directly. Creating deletion mutant collections in other fungal species, including the major human fungal pathogen Candida albicans, will expand our chemical genomics tool set, allowing us to screen for antifungal lead drugs directly. The yeast deletion mutant collection is also being exploited to map large-scale genetic interaction data obtained from genome-wide synthetic lethal screens and the integration of this data with chemical genetic data should provide a powerful system for linking compounds to their target pathway. Extensive application of chemical genetics in yeast has the potential to develop a small molecule inhibitor for the majority of all approximately 6000 yeast genes. PMID- 18416309 TI - Epothilones as lead structures for new anticancer drugs--pharmacology, fermentation, and structure-activity-relationships. AB - Epothilones (Epo's) A and B are naturally occurring microtubule-stabilizers, which inhibit the growth of human cancer cells in vitro at low nM or sub-nM concentrations. In contrast to taxol (paclitaxel, Taxol) epothilones are also active against different types of multidrug-resistant cancer cell lines in vitro and against multidrug-resistant tumors in vivo. Their attractive preclinical profile has made epothilones important lead structures in the search for improved cytotoxic anticancer drugs and Epo B (EPO906, patupilone) is currently undergoing Phase III clinical trials. Numerous synthetic and semisynthetic analogs have been prepared since the absolute stereochemistry of epothilones was first disclosed in mid-1996 and their in vitro biological activity has been determined. Apart from generating a wealth of SAR information, these efforts have led to the identification of at least six compounds (in addition to Epo B), which are currently at various stages of clinical evaluation in humans. The most advanced of these compounds, Epo B lactam BMS-247550 (ixabepilone), has recently obtained FDA approval for the treatment of metastatic and advanced breast cancer. This chapter will first provide a summary of the basic features of the biological profile of Epo B in vitro and in vivo. This will be followed by a review of the processes that have been developed for the fermentative production of Epo B. The main part of the chapter will focus on the most relevant aspects of the epothilone SAR with regard to effects on tubulin polymerization, in vitro antiproliferative activity, and in vivo antitumor activity. Particular emphasis will be placed on work conducted in the authors' own laboratories, but data from other groups will also be included. In a final section, the current status of those epothilone analogs undergoing clinical development will be briefly discussed. PMID- 18416310 TI - Histone deacetylase inhibitors from microorganisms: the Astellas experience. AB - Histone deacetylase (HDAC) inhibitors, such as trichostatin A and trapoxin, which were first found in microorganisms, potently and selectively inhibit HDAC enzymes. They have made a strong contribution to research on HDACs, chromatin control, abnormal epigenetic control in various diseases and the significance of acetylation in posttranslational modification. Recently, HDAC inhibitors have been focused on as potential drugs for the treatment of several diseases, including cancer, although trichostatin A and trapoxin show no effects in animal models because of their metabolic instability in vivo. Chemical modification has been conducted in order to overcome this drawback. We discovered the microbial metabolites FK228 (also known as FR901228, romidepsin, depsipeptide, NSC-630176 and NSC-630176D) and YM753 (spiruchostatin A). Both compounds have bicyclic structures and represent a novel structural class of HDAC inhibitor. The enzyme and tumor cell growth inhibitory activities of FK228 were found to be very potent. It also showed potent HDAC inhibitory activity in vivo. FK228 is the first potent HDAC inhibitor to undergo clinical development as a potential treatment for solid and hematological cancers. Due to its dramatic effect in patients with refractory cutaneous T-cell lymphoma (CTCL), in October 2004 the US Food & Drug Administration (FDA) granted fast-track status to FK228 as monotherapy for the treatment of CTCL in patients who have relapsed following, or become refractory to, another systemic therapy. Thus HDAC inhibitors such as FK228 and YM753 have potential as tools for life science studies and also as therapeutic agents for various intractable diseases. PMID- 18416311 TI - Ascomycete derivative to MS therapeutic: S1P receptor modulator FTY720. AB - Fingolimod (FTY720) represents the first in a new class of immune-modulators whose target is sphingosine-1-phosphate (S1P) receptors. It was first identified by researchers at Kyoto University and Yoshitomi Pharmaceutical as a chemical derivative of the ascomycete metabolite ISP-1 (myriocin). Unlike its natural product parent, FTY720 does not interfere with sphingolipid biosynthesis. Instead, its best characterized mechanism of action upon in vivo phosphorylation, leading to the active principle FTY720-P, is the rapid and reversible inhibition of lymphocyte egress from peripheral lymph nodes. As a consequence of S1P1 receptor internalization, tissue-damaging T-cells can not recirculate and infiltrate sites of inflammation such as the central nervous system (CNS). Furthermore, FTY720-P modulation of S1P receptor signaling also enhances endothelial barrier function. Due to its mode of action, FTY720 effectively prevents transplant rejection and is active in various autoimmune disease models. The most striking efficacy is in the multiple sclerosis (MS) model of experimental autoimmune encephalomyelitis, which has now been confirmed in the clinic. FTY720 demonstrated promising results in Phase II trials and recently entered Phase III in patients with relapsing MS. Emerging evidence suggests that its efficacy in the CNS extends beyond immunomodulation to encompass other aspects of MS pathophysiology, including an influence on the blood-brain-barrier and glial repair mechanisms that could ultimately contribute to restoration of nerve function. FTY720 may represent a potent new therapeutic modality in MS, combined with the benefit of oral administration. PMID- 18416312 TI - Artemisinin--an innovative cornerstone for anti-malaria therapy. AB - Artemisinin-based Combination Therapies (ACT) are recommended by the World Health Organization (WHO) to treat especially multidrug resistant forms of malaria, as currently used medications have become increasingly ineffective. In this chapter, the discovery of artemisinin from Traditional Chinese Medicine and its further development to ACT are reviewed. It is highlighted how the complex supply chain to the naturally occurring endoperoxide artemisinin, required to produce ACT based drugs, was established; thus addressing the significant therapeutic needs and high demands for the medication. PMID- 18416313 TI - [Epidemiology of dog bite lesions in Tierra del Fuego, Argentina]. AB - The coexistence between man and dog has resulted in mutual benefits during thousands of years, nevertheless some problems have recently arisen where bite injuries have an important role. The aim of this work was to describe the epidemiological characteristics of dog bite lesions which occurred during a year in Tierra del Fuego. A descriptive observational, transversal study of patients who were admitted with dog bite injuries to clinic and pediatric emergency services of hospitals and health centers was carried out between 3/1/05 and 3/1/06. A total of 382 records were made. The children group represents 49.5%. The group between 5 and 9 year-old was the most affected (44.4%; IC95% 37.2 51.8). In the adult group, that between 15 and 24 year-old was the most affected (29.5%; IC95% 23.2-36.5). More lesions were registered in men (57.6%) than in women (42.4%) (chi2 = 8.6, p = 0.003). During spring and summer months, 56.5% of the lesions were registered. A 72.8% of the incidents occurred on the public highway. Dogs of big size were responsible of 49.7% (IC95% 44.6-54.9) of the injuries. The 89.8% of the incidents were caused by another person's dog. Of the lesions, 55.8% (IC95% 50.6-60.8) were registered in lower extremities and 11% (IC95% 8.1-14.7) in head and neck. These data show that dog bite lesions affect the health and impact in the quality of life of the population of Tierra del Fuego, Argentina. PMID- 18416314 TI - [Postoperative fever in orthopedic and urologic surgery]. AB - Post-operative fever incidence varies widely. In clean and clean-contaminated surgery the non-infectious fever is more frequent than the infectious fever. We performed a prospective study including 303 patients who underwent orthopedic and urologic elective surgery. The aims of our study were to investigate the incidence of post-operative fever, its etiology, the relationship between time of onset and the etiology, and the usefulness of extensive fever work-up to determine post-operative infection. The incidence of post-operative fever was 14% (42/303) of which 81% (34/42) was noninfectious and 19% (8/42) was infectious. The etiology of the fever in the first 48 hours after surgery was always non infectious (p < 0.001). An extensive fever work-up was performed in patients who presented fever only after the initial 48 hours of surgery with normal physical examination (n = 19) consisting of chest x-ray, blood (2) and urine cultures. The chest x-ray was normal in all the patients, the urine cultures were positive in four cases (21%, IC 95%: 6-45) and the blood cultures in only one case (5%, IC 95%: 0.1-26). Seven patients had post-operative infections without fever as a clinical sign. The most frequently observed etiology of post-operative fever was non-infectious, related to the normal inflammatory host response to surgery. Based on the present results, the extensive fever work-up performed to investigate post-operative infection does not seem to be a useful tool. The diagnosis of post-operative infection was based on clinical follow up and the correct interpretation of the patient's symptoms and signs. PMID- 18416315 TI - [Treatment and post-treatment with lonidamine in human colon carcinoma HT-29 cell line]. AB - Lonidamine (1-[2,4-dichlorophenyl methyl]-1H indazole-3-carboxylic acid), Ind, is an antitumoral drug acting on mitochondria and glucose metabolism. Cell growth and metabolic effects of Ind and drug post-treatment effect were investigated in undifferentiated HT-29 human colonic carcinoma cell line which requires high glucose medium concentration for growth. 0.2 mM Ind significantly decreased cell spreading and growth in monolayer or agar cell culture. After drug treatment cell growth was reestablished to control value within 24 h. Ind modified glycoconjugates and mannose-receptor distribution (analyzed by confocal microscopy), while glucose-glycogen and protein synthesis were not affected, these being the possible reasons for the fast reversible effect. PMID- 18416316 TI - [Tumor size and survival in lung cancer, stage IA]. AB - TNM staging is an important long-term predictor for survival of lung cancer patients. Some studies have shown, however, that tumor size may have intrinsic prognostic value independent of TNM stage. The relationship between tumor size and survival is particularly unclear in T1 tumors. The objective of this study was to assess the prognostic value of tumor size in surgically resected stage I of non-small cell lung cancer (NSCLC). Clinical records of 79 patients with stage IA NSCLC were reviewed. In 34.4% of patients (n = 28) size was < or = 1.5 cm. Surgical mortality was 1.3%. Disease recurrence was noted in 19%. Patients with tumors < or = 15 mm had a significantly higher 5-year survival (95% CI:0.05 vs. 77% CI: 0.07 in > 15mm group). Disease-free survival was 95% for tumors less than 15 mm vs. 72% in larger tumors. Using Cox Multivariate analysis, the most determinant factor for higher risk of mortality was size > 15 mm (relative risk 25.9, IC: 2.3-292, p = 0.004). The independent influence of tumor size in stage IA NSCLC may have practical implications with regards to proposals for screening asymptomatic individuals at high risk for lung cancer. PMID- 18416317 TI - [Alcohol consumption in the emergency room]. AB - We examined patterns of alcohol consumption and blood alcohol levels in emergency room patients. The study was conducted in a public hospital of the province of Jujuy, Argentina. A survey was conducted among all patients during the 24 hours of 4 consecutive days (N = 500) using the Alcohol Use Disorders Identification Test, short version (AUDIT-C). Blood samples were collected in 84% of participants. Results showed that 37% were hazardous drinkers (men 50% vs. women 23%), 29% had symptoms of dependence (men 48% vs. women 11%); 17% of men and 3% of women had high blood alcohol levels, equal or greater than 50 mg/dl. In multivariate logistic regression models men had increased likelihood of hazardous drinking (Adjusted OR 3.2; 95% CI 2.1-5.0), symptoms of dependence (Adjusted OR 7.5; 95% CI 4.4-12.7) and increased blood alcohol levels (Adjusted OR 8.0; 95% CI 2.2-28.8), compared with women. Patients admitted due to accidents, violence, or drug use, had increased likelihood of hazardous drinking (Adjusted OR 2.4; 95% CI 1.5-3.9), dependence symptoms (Adjusted OR 2.0; 95% CI 1.2-3.3) and increased blood alcohol levels (Adjusted OR 7.7; 95% CI 3.0-19.8), compared with those admitted for other reasons. Alcoholism has a significant impact on the occurrence of injuries and on the demand imposed on health services. Prevention programs can potentially reduce the negative health and social impact of alcoholism. PMID- 18416318 TI - Frequency of pancreatic beta-cell autoimmunity markers in patients with autoimmune thyroid disease. AB - A total of 305 ambulatory patients recruited at the Division of Endocrinology, Hospital de Clinicas, University of Buenos Aires, with autoimmune thyroid disease (AITD) were studied to search for associations between autoimmune thyroid disease and presence of serum markers of autoimmune diabetes mellitus. Screening for markers of pancreatic beta-cell autoimmunity was performed by radioligand binding assays (RBA) as follows: autoantibodies to glutamic acid decarboxylase (GADA) and proinsulin (PAA) were determined in all sera, whereas autoantibodies to protein tyrosine phosphatase (IA-2A) and insulin (IAA) were additionally measured in 200 sera randomly selected from the total collection. In addition, every GADA positive serum among the remaining 105 sera was systematically tested for the presence of IA-2A and IAA. In the cohort of 305 AITD patients 22 (7.2%) were previously diagnosed as type 1, type 2 or insulin-requiring type 2 diabetics. Ten of these patients presented serum marker positivity specific for beta-cell autoantigens and 12 were marker negative. On the other hand, considering the majority of non-diabetic AITD patients (n = 283), beta3-cell marker positivity was detected in 17 individuals (6.0%). The prevalence of autoimmune diabetes markers was much higher in the studied population than in the general population utilized as a control group, and GADA was the most frequent marker. PMID- 18416319 TI - [Flow cytometry for the study of mediastinal tumors with abundant lymphoid elements]. AB - The anterior mediastinum is a common site of tumors with abundant lymphoid elements. Flow cytometry is a useful complementary technique to analyze this type of tumors, which provides qualitative and quantitative information. A differential diagnosis can be sometimes made between thymoma and precursor T lymphoblastic lymphoma (T-LBL). Correct identification is of utmost importance for patient treatment. A total of 38 mediastinal tumors were analyzed, and samples were separated for flow cytometry. Flow cytometry data from thymomas and normal thymic tissue were compared with 42 cases of T-LBL from other anatomical locations. Among 38 mediastinal tumors, we found 6 benign lesions, 9 diffuse large B-cell lymphomas (DLBCL), 10 Hodgkin lymphomas (HL), 11 thymomas and 2 T LL. Flow cytometry provided positive information in 24 cases, and defined lymphoid neoplastic cells immunophenotype or the typical lymphocytes accompanying thymomas. Flow cytometry helped differentiate 10 cases of HL and 4 benign lesions from other lymphomas (DLBCL, T-LBL, etc.). CD3, CD4 and CD8 expressions were most useful for the differential diagnosis of thymomas and T-LL. To conclude, flow cytometry is a valid complementary technique, which promptly provides information on mediastinal lesions, requiring small quantities of tissue for both early diagnosis and follow up of these diseases. PMID- 18416320 TI - [Smoking during pregnancy in Argentina and Uruguay]. AB - Argentina and Uruguay are among the countries in which a large proportion of young women smoke. The rate of smokers during pregnancy in both countries is not well known, and data on the characteristics of women who quit smoking during pregnancy compared to those who continue smoking are not available. We conducted a survey including 1512 pregnant women > or = 18 years old (796 in Argentina; 716 in Uruguay), during antenatal visits in public hospitals of large urban regions; 44% of the women in Argentina and 53% in Uruguay had been or were regular smokers. 11% of the surveyed women in Argentina and 18% in Uruguay continued smoking during pregnancy. In both countries, the proportion of women who lived with smokers, allowed smoking at home, and were regularly or always exposed to tobacco smoke indoors, were 49%, 46% and 20% in the subgroup of women who never smoked, 67%, 60% and 32% in those who quit, and 78%, 75% and 52% in those who continued smoking, respectively. The study confirms a serious public health problem in both countries, and documents that environmental exposure persists in subgroups of women, even in those who quit smoking. It is important that the public health sector should provide access to effective programs for smoking cessation, to women who smoke during pregnancy. For the development of a new program, any intervention intending to have at least a moderate and sustainable success, it should seriously consider including components targeting the smoking environment of the pregnant women who smoke. PMID- 18416321 TI - [Young woman with hyperthyroidism associated with severe tricuspid regurgitation]. AB - Cardiovascular manifestations of hyperthyroidism are frequent and sometimes are relevant in the clinical picture. Usually an hyperdynamic circulatory state hallmarks the disease with low peripheral resistance, increased intravascular volume and cardiac output. However, right chamber dilatation with tricuspid valve regurgitation and cardiac failure are unusual. We present the case of a young woman with Graves-Basedow disease without cardiovascular history who complained about palpitations, peripheral edemas, weight loss and fever. The clinical findings were tachycardia with irregular pulse, right heart failure and regurgitant tricuspid murmur that increased with inspiration. The chest radiograph and the echocardiogram showed right ventricular dilatation and severe tricuspid regurgitation without pulmonary hypertension. The treatment with propranolol, corticosteroids and diuretics was successful. The patient was asymptomatic with sinus rhythm. We discuss the effects of thyroidal hormones on the cardiovascular system and postulate pathophysiologic mechanisms of heart failure in hyperthyroidism. PMID- 18416322 TI - [Amniotic fluid embolism. Diagnostic criteria in two fatal cases]. AB - Amniotic fluid embolism still remains an important cause of maternal mortality. We present information obtained by echocardiography and right cardiac catheterization of two patients who developed amniotic fluid embolism and died from shock and disseminated intravascular coagulation despite intensive medical treatment. Although the pathophysiology remains controversial, amniotic fluid embolism can be presumptively diagnosed and managed with hemodynamic values and echocardiography. PMID- 18416323 TI - [Real time intracardiac electrograms for the diagnosis of pacemaker malfunction]. AB - Chronic ventricular lead dislodgement is an infrequent complication of pacemaker implantation. Occasionally, the dislodged lead may sense and capture a chamber in which the lead was not originally positioned. Intracardiac real time electrograms and channel markers are useful tools for the diagnosis of pacemaker malfunction. We present the case of a patient with a ventricular lead dislodgement into the atrium. The ventricular lead was able to sense and capture the atrium. Initial diagnosis was performed based on the deductive analysis of intracardiac real time electrograms and channel markers and confirmed by chest X-ray. PMID- 18416324 TI - [Betalactam antibiotics combined with bectalactamases inhibitors. Amoxicillin sulbactam]. AB - Betalactamases production is one of the main bacterial resistance mechanisms to betalactam antibiotics. The use of bectalactamases inhibitors combined with betalactam antibiotics allows the inactivation of certain betalactamases produced by Gram positive, Gram negative and anaerobic organisms, and even by mycobacteria. Betalactamases inhibitors are an improved therapeutic alternative compared with the other betalactam since, in most cases, they cover a wider antimicrobial spectrum than their analogues. Betalactamases enzimatic activity is specifically directed to the betalactam ring hydrolisis, producing a compound without antibacterial activity. According to their genomic position within microorganisms, betalactamases can be either chromosomic or plasmidic. Currently there are three betalactamases inhibitors locally available: clavulanic acid, sulbactam and tazobactam. Of them, only sulbactam has an intrinsic antimicrobial activity against penicillin binding proteins. The clinical experience from over 20 years confirms that the combination of betalactam antibiotics is effective in the empirical initial treatment of respiratory, intraabdominal, urinary tract and gynecologic infections, including those of polymicrobial origin. In the specific case of amoxicillin-sulbactam, experiences have shown the effectiveness of the combination in the treatment of peritonsillar abscess, otitis media, sinusitis, community acquired pneumonia, acute exacerbation of chronic obstructive pulmonar disease (COPD), urinary tract infection and obstetric/gynecologic infections. The spectrum and pharmacologic properties of this combination makes it also an excellent option for the treatment of skin/soft tissue and intraabdominal infections. PMID- 18416325 TI - [Argentine Consensus of Congenital Toxoplasmosis]. AB - The mother-to-child transmission in Toxoplasma gondii infection occurs only when the infection is acquired for the first time during pregnancy. The prenatal and early postnatal diagnosis can only be achieved by serological testing. Serologic tests have different sensitivities, specificities and complexities, so that different tests in more than one blood sample are necessary for the diagnosis. Serological follow-up of the infants should be conducted during the first year of life or until the diagnosis of congenital toxoplasmosis can be ruled out. Treatment recommendations try to reduce the transmission rate and the risk of congenital damage. Congenital toxoplasmosis incidence rate is approximately 5 per 1000 births, but can be reduced to 0.5 per 1000 with an active screening program. The aim of this consensus group was to review the scientific literature on congenital toxoplasmosis and prepare a statement on prevention, diagnosis and treatment that should be implemented in our country. PMID- 18416326 TI - [Constructive feedback. A strategy to enhance learning]. AB - Constructive feedback is defined as the act of giving information to a student or resident through the description of their performance in an observed clinical situation. The following keys are required during feedback in order to get student's performance improvement: the observation of an event, the instructor's comment on said performance, by following always a pre-established pattern or standard, and advice on improvement. Major impact takes place when a student or a resident compares the teacher's feedback to its own performance. The dissonance between desired and actual performance constitutes a strong motivation and deep learning generator. Most of the teachers find it essential, but in general they carry it out in a short and non specific way. Medical students and residents are willing to get and need constructive feedback and they clearly associate it to high-quality learning. However, even though they appreciate it, they seldom request it. It is essential to take maximum advantage of feedback by giving recommendations, producing reactions and self-analysis and also by defining an appropriate action plan. It takes time and effort to apply this tool, but it is of great value for the growth and academic development of students and residents. PMID- 18416327 TI - [Nobel Prize 2007 in Physiology or Medicine. Designing genomes in mice]. PMID- 18416328 TI - [Preventive medicine in the elderly. Yes or No?]. PMID- 18416329 TI - [Cryptomnesia]. PMID- 18416330 TI - [Generational transformation of biomedical investigation]. PMID- 18416331 TI - Primary cutaneous leiomyosarcoma: a histological and immunohistochemical study of 4 cases. AB - Primary cutaneous leiomyosarcoma is an uncommon malignant neoplasm with a predilection for the lower extremities. A retrospective study of 4 cases was undertaken to analyse the clinicopathological characteristics and immunohistochemical profile of these neoplasms with emphasis on prognosis. Two male and 2 female patients aged between 49 and 80 years presented with painless tumours involving the lower lip, the chin, the scrotum and the shoulder. Histological examination of the initial biopsy specimen established a diagnosis of cutaneous leiomyosarcoma. All cases co-expressed smooth muscle actin and vimentin regardless of primary tumour site. Wide surgical excision of the tumour was performed in only 3 cases, and the remaining patient refused further treatment. Of the patients undergoing surgical intervention, local recurrence occurred in one case. No metastases were observed. Long-term follow-up of patients with cutaneous leiomyosarcoma is mandatory to detect local recurrence and distant metastases that can occur even years after the initial excision. PMID- 18416332 TI - [Budget management in anatomical pathology: health technology assessment of new methodologies]. AB - The author's experience in heath technology assessment of new methodologies for routine diagnosis at the Department of Anatomical Pathology at the Trento Hospital is presented. The workload of the department together with the annual budget trends (from 2000 to 2006) of the various costs is analysed. Budget analysis also allows evaluation of expenses relative to the introduction of new tests, which are increasingly requested in order to personalise therapy accordingly to the biological profile of individual patients. Health Technology Assessment permits in-depth analysis of the efficacy, safety, costs, benefits and feasibility in addition to providing a measurement of the contribution to improving the quality of work and life. This is an important tool in decision making processes for pathologists, especially in consideration of the limited resources available in healthcare. PMID- 18416333 TI - [Choroid plexus tumours: cytogenetic analysis of a single case and literature review]. AB - Choroid plexus tumours are intraventricular papillary lesions that are observed in typical papillomas, atypical papillomas and carcinomas. They usually occur in childhood, and can result in hydrocephalus and increased intracranial pressure. The present paper describes a case of choroid plexus papilloma in an adult woman; cytogenetic analysis of the lesion is also presented, which demonstrated the presence of tumour mosaicism. Compared to the chromosomal aberrations observed in previous cases, those in the present tumour do not show significant differences between papillomas harboured in adults and paediatric patients. Moreover, there was no apparent correlation between genetic alternations in typical and atypical papillomas and prognosis or recurrence. PMID- 18416334 TI - Breast metastases from undifferentiated nasopharyngeal carcinoma. AB - The most common primary tumours metastasising to the breast include melanoma, lymphoma, lung cancer and ovarian cancer, while metastases from head and neck cancer are rare. Metastases from undifferentiated nasopharyngeal carcinoma cancers are extremely rare, and only 3 well-documented cases have been reported in the English literature. We report a fourth case of breast metastases from nasopharyngeal carcinoma confirmed by in situ hybridization, focusing on clinical data as well as radiologic and pathologic features. PMID- 18416335 TI - Small cell malignant melanoma: an unusual morphologic variant. AB - Small cell melanoma is a recognized rare variant of malignant melanoma. We report a case of a brown, ulcerated papule on the left third finger of an 80-year-old woman. Microscopic examination revealed the presence within the epidermis of diffuse sheets of monomorphic small to medium-sized cells. The nuclei were round or oval, and hyperchromatic with inconspicuous nucleoli. Melanin pigment was either absent or minimal. This case report draws attention to the difficulties encountered in the histological diagnosis of this rare variant of malignant melanoma. PMID- 18416336 TI - [Mammary hamartoma with atypical stromal cells: a potential diagnostic dilemma]. AB - Hamartoma of the breast is a pseudotumoural lesion that does not usually pose diagnostic problems for the pathologist. Although atypical stromal cell (ASCs) can be encountered in several benign and malignant breast lesions, their occurrence in hamartoma has not been reported to date. The authors report a case of breast hamartoma containing numerous atypical mono- or multinucleated stromal cells within the fibro-fatty component. This unusual feature raised differential diagnostic problems with pleomorphic lipoma, well-differentiated liposarcoma and malignant phylloid tumour with a lipomatous heterologous component. Immunohistochemistry, showing positivity to vimentin and CD34, revealed that ASCs are fibroblastic in nature, and thus are likely to represent a morphological variant of the fibroblasts of the native mammary stroma. PMID- 18416337 TI - [Angiomyofibroblastoma of the vulva: a rare but distinct entity. Case report and literature review]. AB - Angiomyofibroblastoma is a benign vulvar tumour involving soft tissue that is characterized by alternating hypocellular and hypercellular areas of spindle stromal cells, admixed and aggregated around blood vessels. It is important to recognize this entity as it shows benign behaviour with respect to other mesenchymal tumours of the vagina, which have a more aggressive behaviour. PMID- 18416338 TI - Mesenchymal hamartoma of the chest wall in an infant. AB - Mesenchymal hamartoma of the chest wall is a very rare, benign tumour with distinct clinical, radiological and histopathologic characteristics. The lesion develops during foetal life, and is present at or shortly after birth with an extrapleural mass arising from the rib cage with or without respiratory distress and marked rib deformity. Several imaging techniques have been used for diagnosis, but a definitive diagnosis is established only by histopathological examination. Such lesions are composed of a varying admixture of hyaline cartilage that has features resembling growth plate cartilage, along with fascicles of spindle cells, woven bone and hemorrhagic cysts. Accurate diagnosis of mesenchymal hamartoma is important since many chest wall masses in children are malignant. We report a case of mesenchymal hamartoma of the left posterior chest wall surgically resected in an infant who was found to have a palpable mass at birth. Two years after surgery, the patient is alive and well, with no evidence of recurrence. PMID- 18416339 TI - Granuloma multiforme: first case report in Tunisia. AB - Granuloma multiforme is a rare granulomatous skin disease, usually reported in sub-Saharan African countries. The exact aetiology of granuloma multiforme is still unknown. We report the case of a patient who presented clinical and histopathological features of granuloma multiforme that can be considered the first described case in Tunisia. PMID- 18416340 TI - Cystic nephroma in the adult: pathological aspects and therapeutic implications. AB - Cystic nephroma is a benign renal neoplasm. Since its initial description, there has been much debate regarding its origin. Preoperative diagnosis of Cystic nephroma is difficult to achieve. The differential diagnoses of Cystic nephroma are recently described mixed epithelial and stromal tumours of the kidney and cystic renal cell carcinoma. The Authors report three cases of Cystic nephroma and illustrate the clinical, radiological and histological features of this renal neoplasm. PMID- 18416342 TI - Neoteric therapies in alcoholic hepatitis--has the outcome changed? PMID- 18416341 TI - [New diagnostic markers for mesothelioma]. PMID- 18416343 TI - Update in liver imaging. AB - Over the last decade, liver imaging has experienced a revolution providing a bewildering array of options for detection and characterisation of liver lesions. Cross-sectional imaging modalities like computed tomography and magnetic resonance imaging have improved in speed and resolution, thereby facilitating multiphasic scanning of the liver. The advent of the use of contrast agents for ultrasonography and magnetic resonance imaging has further paved the way for definitive diagnosis in an attempt to obviate the need for invasive diagnosis. This article captures advances made in ultrasonography, computed tomography, magnetic resonance imaging and positron emission tomography and brings the specialist up to date with the latest in liver imaging. PMID- 18416344 TI - Abdominal compartment syndrome--an old syndrome, a new perspective. AB - The abdominal compartment syndrome is a life threatening condition resulting from pathologic elevation of the intraabdominal pressure. Prompt diagnosis is required to avoid significant sequelae. Diagnosis of this syndrome is based on clinical findings and intra abdominal pressure monitoring. Treatment consists of decompressive laparotomy, which corrects the pathology. Various surgical techniques are described to manage the open abdomen. Despite considerable attention accorded to this disorder, it is still associated with high morbidity and mortality. This review article deals with the identification of risk factors, pathophysiology, diagnostic criteria and treatment of critically ill patients with the abdominal compartment syndrome. PMID- 18416345 TI - Small intestinal bacterial overgrowth in North Indian patients with celiac disease. AB - BACKGROUND: Occasionally celiac patients continue to experience gastro-intestinal symptoms even with a gluten free diet. In these cases, small intestinal bacterial overgrowth may be one of the causes of the lack of response. Therefore, this prospective study was planned to determine the prevalence of small intestinal bacterial overgrowth in celiac patients. MATERIALS AND METHODS: We studied 87 confirmed cases of celiac disease from North India and 87 age and sex matched controls. Celiac disease was confirmed by positive IgA antitissue transglutaminase on ELISA. 80 g glucose hydrogen breath test (non-invasive test) was performed to establish small intestinal bacterial overgrowth. Rise of more than 10 ppm in hydrogen concentration over baseline value within two hours was considered suggestive of small intestinal bacterial overgrowth. RESULTS: Out of 87 patients with celiac disease, 49 were male and 38 were female. The mean (+/ SD) age for male patients was 26.3 +/- 16.3 years (range 14-59 years) and for female patients was 28.4 +/- 15.6 years (range 16-58 years). Amongst the controls, 52 (59.8%) were male and 35 (40.2%) were female. The mean (+/- SD) age for male controls was 27.6 +/- 14.5 years (range 15-57 years) and for female controls was 29.3 +/- 16.5 years (range 18-59 years). Hydrogen breath test was suggestive of bacterial overgrowth in 18 of the 87 (20.7%) celiac disease patients but not in any of the apparently healthy controls. CONCLUSION: This study indicates that a large number of celiac patients from North India suffer from bacterial overgrowth which can be accordingly treated with antibiotics. PMID- 18416346 TI - Is conventional sphincterotomy possible in patients with common bile duct stones impacted at the ampulla of Vater? AB - BACKGROUND: Needle knife precut papillotomy has been used to gain access or remove impacted common bile duct stones at the ampulla of Vater. However, precut papillotomy is a risky procedure with high complication rates and is presumably best dealt with by the expert. We attempted to find out the feasibility of performing conventional endoscopic sphincterotomy in patients with impacted stones at the ampulla. METHODS: All patients undergoing endoscopic retrograde cholangiopancreatography for stones impacted at the ampulla between the years 2000 and 2005 were enrolled in the study. All attempts were made to achieve biliary cannulation through the conventional route including use of the long route or guidewire to cannulate the papilla. Needle knife papillotomy was resorted to only if attempts to cannulate the papilla through the conventional route failed. The success and complications of the procedures were recorded. RESULTS: Thirty-two patients were enrolled. The impacted bile duct stone could be removed in all the patients. Conventional endoscopic retrograde cholangiopancreatography and endoscopic sphincterotomy were performed in 23 patients with impacted stones. The long route had to be employed in seven patients while guidewire cannulation of the papilla was needed in four patients. Needle knife papillotomy was needed in only nine patients. None of the patients in whom the conventional route was employed developed complications, whereas bleed requiring endoscopic management was observed in three (33%) of the nine patients in whom needle knife papillotomy was performed. CONCLUSIONS: Endoscopic retrograde cholangiopancreatography and endoscopic sphincterotomy can be performed by the conventional route in the majority of patients with impacted stones at the ampulla. This approach appears to be safer than needle knife papillotomy. However, needle knife papillotomy may be required in patients in whom biliary access cannot be gained through the conventional approach. PMID- 18416347 TI - Serum pepsinogen I and II levels in various gastric disorders with special reference to their use as a screening test for carcinoma stomach. AB - INTRODUCTION: The role of serum pepsinogen in the diagnosis of gastric carcinoma is well established. Its role in other common upper alimentary disorders has not been widely studied. The aim of this study was to describe the effect of various gastric disorders on the levels of pepsinogen I, pepsinogen II and pepsinogen I/II ratio, with an emphasis on the diagnosis of carcinoma stomach in the South Indian population. METHODS: A total of 210 patients in seven groups, including one control group, were studied. The groups included patients with carcinoma stomach, Helicobacter pylori gastritis, peptic ulcer, portal hypertensive gastropathy, non-ulcer dyspepsia and erosive gastritis. Serum pepsinogen I, pepsinogen II and pepsinogen I/II ratio were estimated using an enzyme-linked immunosorbent assay technique. RESULTS: Patients with carcinoma of the stomach, when compared with controls, had a significantly lower pepsinogen I level (87.2 microg/L vs. 158.1 microg/L, p=0.0002) and pepsinogen I/II ratio (4.3 vs. 7.2, p = 0.0001). No significant change in pepsinogen levels occurred in the other groups. The cut-off levels of pepsinogen I (115.3 microg/L) and pepsinogen I/II ratio (6.2), determined by THE ROC curve, when applied in parallel provided a sensitivity of 97% and a negative predictive value of 91.4% for the diagnosis of carcinoma stomach. When the tests were applied in parallel, the likelihood ratio of a negative test was 0.06, indicating that individuals without carcinoma stomach were 16 times more likely to have a negative test than those with carcinoma. This fulfilled the essential prerequisites of an ideal screening test. CONCLUSION: Serum pepsinogen estimation is a useful diagnostic tool in the diagnosis of carcinoma stomach. The significance of serum pepsinogen level in portal hypertensive gastropathy, non-ulcer dyspepsia, peptic ulcer, Helicobacter pylori gastritis and erosive gastritis was not established. PMID- 18416348 TI - Quality of life and nutritional status in alcohol addicts and patients with chronic liver disease. AB - OBJECTIVE: The aim of the study was to assess and compare the nutritional status and quality of life in chronic liver disease (alcoholic and non-alcoholic) patients and alcohol addicts. METHODS: Patients with alcoholic liver disease (n=41), nonalcoholic liver disease (n=40), alcohol addicts (n=25) without liver disease and healthy controls (n=25) were randomly selected. Nutritional status was assessed using anthropometric measurements viz. skin fold thickness, arm muscle circumference and area. Biochemical estimations included liver function tests. Food intake was assessed using 48 hour recall and macro-nutrient intake was calculated. Quality of life was assessed using the SF-36 questionnaire. RESULTS: The mean value of mid-arm muscle area was significantly lower in patients from the non-alcoholic liver disease group when compared with the other 2 groups (p= 0.0). Body fat store depletion was significantly lower in the alcohol addict group when set against the alcoholic liver disease and non alcoholic liver disease groups (p= 0.0). The mean percentages of ideal calories (p= 0.0) and proteins (p= 0.0) were significantly higher in alcohol addicts but no significant differences in the mean percentage of fat intake (p= 0.1) was observed. The frequency of macro-nutrient deficiency was highest in the non alcoholic liver disease group (p= 0.0). Ethanol consumption was not significantly different between alcohol addicts and patients suffering from alcoholic liver disease (p=0.06). Patients with liver disease (irrespective of aetiology) scored significantly lower on the quality of life scale when compared to alcohol addicts. CONCLUSIONS: Malnutrition is more frequent and severe in patients suffering from chronic liver disease in comparison to alcohol addicts. The health status is significantly poorer in patients suffering from alcoholic liver disease. Alcohol does not seem to play a primary role in the pathogenesis of liver disease and malnutrition. PMID- 18416349 TI - Severe and treatment resistant strongyloidiasis--indicator of HTLV-I infection. AB - The association between severe and persistent strongyloidiasis with human T cell lymphotropic virus type I (HTLV-1) infection is well documented in reports from HTLV-1 endemic regions like Japan and Jamaica. But there are no reports from non endemic areas like India. We report a case of severe intestinal strongyloidiasis in a 45-year old Keralite man, living in Sikkim. Despite standard treatment with many courses of albendazole, his stool persistently showed Strongyloides stercoralis larvae. In the absence of other immunosuppressive conditions, human T cell lymphotropic virus type I infection was considered and determined positive. Subsequently, treatment with 2 courses of ivermectin achieved eradication of the infection. On follow-up, 3 years later, his stools again revealed Strongyloides stercoralis larvae. PMID- 18416350 TI - Primary oesophageal tuberculosis: a rare entity. AB - Tuberculous infection of the oesophagus is rare and primary oesophageal tuberculosis is seen even more infrequently. We report a case of oesophageal tuberculosis in a 32-year-old female patient who presented to us with odynophagia and weight loss. Endoscopy showed a solitary ulcerative oesophageal lesion. Further investigation resulted in a diagnosis of oesophageal tuberculosis with no manifestations of tuberculosis elsewhere. She responded well to antitubercular treatment. This case was classified as primary oesophageal tuberculosis. PMID- 18416351 TI - Enteric fever with suspected intestinal perforation and incidental renal cell carcinoma--a case report. AB - Enteric fever presents with protean manifestations, at times eluding the treating physician. We report the case of a 19-year-old woman whose clinical presentation suggested enteric fever, however, autopsy revealed occult renal cell carcinoma. We emphasise here, the need to investigate non-infective causes of pyrexia. PMID- 18416352 TI - Aggressive angiomyxoma of the jejunum: a rare cause of obscure gastrointestinal bleeding. AB - Obscure gastrointestinal bleeding accounts for nearly 5% of all gastrointestinal haemorrhage and is frequently due to a lesion in the small bowel. We report the case of a male patient with obscure overt gastrointestinal bleed in whom repeated upper gastrointestinal endoscopy, colonoscopy, computed tomography scanning and exploratory laparotomy showed no specific pathology. However, on capsule endoscopy done subsequently, a small polyp in the jejunum was located and resected. Histology revealed an aggressive angiomyxoma. This type of small bowel lesion has not been reported in the literature before. PMID- 18416353 TI - Benign recurrent intrahepatic cholestasis (BRIC) in an adult. AB - Benign recurrent intrahepatic cholestasis is a rare hereditary disorder characterised by recurrent episodes ofcholestasis. We report the case of a young male patient with benign recurrent intrahepatic cholestasis who presented to us with recurrent cholestatic jaundice and pruritus with negative work up for all possible aetiologies and a liver biopsy consistent with intrahepatic cholestasis. He improved on treatment with ursodeoxycholic acid and ondansterone and is doing well on follow up. PMID- 18416354 TI - Jejunogastric intussusception--a rare cause of recurrent upper gastrointestinal bleeding: case report and review of literature. AB - Jejunogastric intussusception is a known complication following gastrojejunostomy. However, only occasionally have they presented with haematemesis and we are unaware of any case where it led to recurrent haematemesis. We describe a case where the patient developed recurrent upper gastrointestinal bleeding due to recurrent episodes of jejunogastric intussusception following gastrojejunostomy performed 12 years earlier for duodenal ulcer disease. PMID- 18416355 TI - Pancreatic mucinous cystadenoma. AB - We report here the case of a 75-year-old lady who presented to us with a 4-month history of abdominal symptoms. The computed tomography scan revealed a cystic lesion in the tail of the pancreas. Distal pancreatectomy was done and biopsy showed a benign mucinous neoplasm. Because this is potentially malignant it is vital to diagnose it before it becomes malignant; identification of this entity remains a diagnostic and therapeutic challenge. PMID- 18416356 TI - Partners in critical care. AB - Patient- and family-centred care (PFCC) concepts are increasingly cited in the critical care literature and are a welcome addition to the vernacular of the intensive care unit (ICU). The implementation and maintenance of a supportive PFCC environment is challenging, however, and usual strategies for knowledge translation using guidelines and policies, no matter how articulate, have not yet resulted in sustained practice change at the point of care delivery. In this article, co-authored by community partners, the physician director and nurse leader of one tertiary care ICU, we describe an initiative in which patient and family representatives were included in the ICU interdisciplinary team membership. After two years and now, at the conclusion of the assignment, options for community partner participation in various activities related to unit governance are shared. PMID- 18416357 TI - Exploring CRRT practices in ICU: a survey of Canadian hospitals. AB - BACKGROUND: Continuous renal replacement therapy (CRRT) is a highly specialized therapy not only for patients with acute renal failure, but also for patients with other critical conditions. The nursing work involved in CRRT is complex and the learning requirements are challenging to sustain ongoing competence. There are also adverse events associated with CRRT, such as those resulting from the anticoagulation therapy. Limited nursing literature is available regarding the use of CRRT by nurses in Canadian intensive care units. METHODS: In 2005, the authors conducted a national survey of CRRT nursing practices with nurse educators in intensive care units. The survey had fixed choice, as well as open ended questions exploring various aspects of CRRT implementation, including education provided and adverse events experienced by patients. RESULTS: Of the 53 teaching and non-teaching hospitals that were sent the survey, 50 replied (94% response rate). Thirty-four of the sites used CRRT. The results reported here represent the findings from the survey questions pertaining to education and adverse events only. CONCLUSION: The results indicate that education, continuing competence, and prevention and management of adverse events, such as bleeding and filter clotting, are the major issues related to CRRT nursing practice across Canada. PMID- 18416358 TI - Implementing an ICU outreach team model. AB - There is an increasing need for critical care expertise outside of the intensive care unit (ICU). This is particularly related to high acuity levels and limited nursing resources. Teams of critical care health care providers have been formed to meet this need. Team structure and services provided vary according to the organization's need and the resources available. In this article, the process of establishing an ICU outreach team without additional resources, in a 200-bed tertiary care hospital in central Saskatchewan is presented. The team responds to calls from within the hospital to provide critical care expertise to patients about whom the professional caregiver is concerned, or patients who have early warning signs of physiological distress. Implementation of the team has been successful in improving timely patient care, providing a resource for nurses to access for their patients, and providing an opportunity for education and support for nurses on inpatient units. PMID- 18416359 TI - The Canadian ICU collaborative: working together to improve patient outcomes as an interprofessional team. PMID- 18416360 TI - Preventing substitution errors involving high-concentration heparin products. AB - Critical care practitioners routinely administer heparin for various indications (e.g., treatment of acute coronary syndrome, venous thromboembolism prophylaxis, line maintenance) and by various routes (e.g., intravenously, subcutaneously). Knowledge of reported incidents involving high-concentration heparin products can increase practitioner awareness of risks for error-induced injury associated with storage and administration of anticoagulants, such as heparin. Substitution errors leading to administration of an incorrect dose of unfractionated heparin are highlighted and suggestions for system-based error prevention strategies are provided. PMID- 18416361 TI - WIPHL helps doctors empower patients. PMID- 18416362 TI - Reflections on a summer internship in public health. PMID- 18416363 TI - University of Wisconsin School of Medicine and Public Health: a century of inspiration. PMID- 18416364 TI - Chronic kidney disease (CKD) in Wisconsin: time to address this public health problem. PMID- 18416365 TI - Wisconsin firearm deer hunting season: injuries at a level I trauma center, 1999 2004. AB - CONTEXT: Hunting continues to be a passion and common pastime for many US citizens, especially in rural areas. Unfortunately, with the large volume of hunters entering the woods each season, hunting injuries continue to be common. OBJECTIVE: Review the experience of a level I trauma center during each of Wisconsin's 9-day deer firearm hunting seasons over a 6-year period and identify potential prevention elements based on the findings. DESIGN: We retrospectively reviewed all hunting-related injury patient data entered into the University of Wisconsin Hospital and Clinic's (UWHC) level I trauma registry from 1999 to 2004, for each 9-day Wisconsin deer hunting firearm season. We compared injury occurrence with Wisconsin DNR statewide hunting-related firearm injury incidence data over the same time frame. The study was conducted at a level I university tertiary referral trauma center. The study included any patient admitted to the UWHC during the study period with a hunting-related injury entered into the trauma registry. OUTCOMES MEASURED: Primary outcomes recorded included patient demographics, mechanism of injury, types of injuries, comorbidities, injury severity scores, and mortality. RESULTS: Twenty-four patients were treated for hunting-related injuries during the study period. The majority of hunters were male (95%), with an average age of 44.5 years. Treestand injuries accounted for 16 of the 24 injuries treated; the rest of the injuries were firearm-related. Most of the injuries (18) occurred during the first 3 days of the hunting season, with the remaining 6 injuries taking place around the Thanksgiving holiday period. Injury severity scores (ISS) ranged from 1 to 50. Orthopedic concerns accounted for 79% of the injuries, while general surgical was 50%, and neurosurgical was 12.5%. Two fatalities occurred due to complications from injuries caused by falling from a treestand. CONCLUSION: Falls from a tree-stand and firearm shootings represent 2 mechanisms for severe hunting-related injuries during the 9-day deer firearm hunting season in Wisconsin. Formal treestand safety instruction should be emphasized during hunting education classes and all hunters need to heed safe hunting recommendations pertaining to firearms and treestands, and be particularly cautious during the first few days of the hunting season. PMID- 18416366 TI - Local public health department funding: trends over time and relationship to health outcomes. AB - BACKGROUND: Local health departments contribute to population health improvement through the core functions of assessment, policy development, and assurance. Their capacity to perform these functions may be affected by funding and staffing. OBJECTIVE: To describe local health department funding and staffing levels and determine the relationship between these measures and county-level health outcomes. METHODS: Ten years of total funding, funding by revenue source, and staffing data from local health departments in all 72 Wisconsin counties were collected from the Department of Health and Family Services and analyzed. Summary measures for county health outcomes were obtained from the 2006 Wisconsin County Health Rankings, and a correlation matrix was created to determine associations between outcomes and measures of health department capacity. RESULTS: On average, Wisconsin local health departments spend $20.60 per capita, ranging from $7.50 to $68.30 among counties. While total per capita funding in the state (adjusted for inflation) increased $0.82 per year, a closer look reveals 3 distinct periods: increases of $0.20 per year during 1995-1997 and $1.33 per year during 1997-2001; but a decrease of $0.27 during 2001-2004. Local health departments in counties with worse health outcomes had only slightly higher average funding and staffing levels during 2002-2004. CONCLUSION: Levels of health department funding in Wisconsin, already low by US standards, declined slightly in the past 3 years. Although counties with the worst health outcomes had slightly higher levels of public health funding, considerable disparities exist. State policymakers might consider investing more resources in counties with the greatest need, to support evidence-based public health programs and reduce existing geographic health disparities in Wisconsin. PMID- 18416367 TI - Improving the accuracy of payment classifications through use of a case management protocol. AB - OBJECTIVES: The purpose of this study is to determine if a Case Management Protocol (CMP) improves the accuracy of assignment of Medicare patients to the appropriate payment classification. METHODS: MetaStar, Wisconsin's Quality Improvement Organization (QIO), invited Wisconsin hospitals to participate in this project; 19 hospitals did so. A CMP enables physicians to enter an order in the medical record to "admit the patient under the case management protocol" when it is not obvious to the physician whether the patient should be admitted as an inpatient or placed in an outpatient status. A trained case management professional accesses the documentation in the medical record and makes a recommendation to the physician as to the appropriate status. The decision is ratified by the physician in the form of a signed order. RESULTS: In comparing 1 day inpatient stays as a percentage of all hospital stays in a group of hospitals that considered the use of the CMP, to that same percentage in the hospitals that did not consider the use of a CMP, there was a reduction of 1-day stays for the former group that was significantly (P<.01) greater than for the latter group; the decrease in target payments for the former group also was significantly greater than that for the latter group (P<.01). CONCLUSION: The use of a CMP to assign Medicare patients to appropriate payment classifications is an effective method of increasing the accuracy of such assignment. PMID- 18416368 TI - Spinal intramedullary cysticercosis of the conus medullaris. AB - Neurocysticercosis is the most common central nervous system (CNS) parasitic disease worldwide, but spinal cysticercal infection is relatively rare, especially in the United States. Because of increased immigration to the United States from endemic areas, the incidence of neurocysticercosis has risen, especially in California, Texas, Arizona, and other southwestern states, but not in Wisconsin. Spinal intramedullary cysticercosis involving the conus medullaris is an uncommon clinical condition that can lead to irreversible neurological deficits if untreated. Rarely, Taenia solium, a cestode that causes neurocysticercosis, may produce spinal intramedullary lesion, which may mimic an intramedullary tumor. We report a case of thoracolumbar spinal intramedullary cysticercosis caused by Taenia solium. Spinal neurocysticercosis should be kept in mind in the differential diagnosis of intramedullary conus lesions even if the patient lives in Wisconsin. PMID- 18416369 TI - Amyloidosis presenting as lower gastrointestinal hemorrhage. AB - AL-Amyloid rarely presents in the gastrointestinal tract as acute gastrointestinal hemorrhage, especially in the absence of clinical disease elsewhere in the body. There are no reported cases of monoclonal gammopathy of undetermined significance progressing to AL-Amyloid presenting as lower gastrointestinal hemorrhage. We report a case of a patient initially diagnosed with monoclonal gammopathy of undetermined significance who progressed to AL Amyloid over the course of 1 year. His progression resulted in primary colonic amyloidosis that manifested as lower gastrointestinal hemorrhage. The diagnosis was made by biopsy of a sigmoid plaque demonstrating necrotic material on histopathology. Amyloid deposition was seen on congo red and on birefringence. The bleeding stopped spontaneously without intervention and he was discharged his fourth day in the hospital. Further evaluation revealed no involvement in other organ systems. The plan is to treat with melphalan and dexamethasone. We conclude that early endoscopic examination and biopsy of the surrounding intestinal tissue is indicated when patients with monoclonal gammopathy of undetermined significance present with gastrointestinal hemorrhage to evaluate for the progression to AL-Amyloidosis. Treatment to prevent recurrent hemorrhage and further progression of the disease should be considered. PMID- 18416370 TI - Could declining home values affect your retirement plans? PMID- 18416371 TI - [A pioneer in pain management. Interview by Sylvie Warnet]. PMID- 18416375 TI - [Orienting care to a nursing perspective]. PMID- 18416376 TI - [Haptonomy and care of the elderly patient]. PMID- 18416377 TI - [Value of haptonomy in neonatal intensive care]. PMID- 18416378 TI - [The corporal approach in care]. PMID- 18416379 TI - [To alleviate and reconstruct by sensory comunication]. PMID- 18416380 TI - [Caring in rehabilitating the body image of burned patients]. PMID- 18416381 TI - [Caring in another way, the health promotion space in the Limoges university hospital center]. PMID- 18416382 TI - [Quality of life promotion, a part in the whole care process]. PMID- 18416383 TI - [Taking care until the end of life]. PMID- 18416384 TI - [Do you know about gypsotherapy?]. PMID- 18416385 TI - [The nursing care record]. PMID- 18416386 TI - [Multidisciplinary management of chronic non-cancerous pain]. PMID- 18416387 TI - [Analysis of therapeutic management five years after a cerebrovascular accident]. PMID- 18416388 TI - [A real case in gastrointestinal surgery]. PMID- 18416389 TI - [Oral hygiene in the hospital milieu]. PMID- 18416390 TI - [Jean-Baptiste Pussin (1745-1811)]. PMID- 18416391 TI - [Cytological and bacteriological examination of urine]. PMID- 18416392 TI - [New drugs for HIV infection]. PMID- 18416394 TI - Why mandatory training needs resuscitating. PMID- 18416393 TI - Could Colin Norris have been stopped? PMID- 18416395 TI - 'We had a vision of a better service'. Interview by Clare Lomas. PMID- 18416396 TI - New guidance on osteoarthritis focuses on patient education. PMID- 18416397 TI - Patient assessment. Part 4 - blood glucose testing. PMID- 18416398 TI - Communication in end-of-life cardiac care 1: difficult issues. AB - This is a two-part unit on communication issues in end-stage cardiac disease. Part 1 discusses the main communication problems relevant to end-stage heart failure. Patients may wish to discuss end-of-life issues but are denied the chance to do so as nursing staff often lack the skills or confidence in dealing with such issues. PMID- 18416400 TI - A chance to make a difference. PMID- 18416399 TI - Restructuring trauma wards to reduce patient stay. PMID- 18416401 TI - Only part of the solution. PMID- 18416402 TI - The 372 million pound sterling diet. PMID- 18416403 TI - Images of obesity. PMID- 18416404 TI - Wellbeing of new mothers. AB - This paper reports indicators of wellbeing and early parenting stress in a representative sample of first-time mothers in north-east England. A total of 185 mothers were recruited in the antenatal period to a controlled trial of an early parenting intervention. They were interviewed at home when the baby was aged around one month, and completed the General Health Questionnaire (GHQ12) and the Parenting Daily Hassles Scale. Almost half of mothers reported psychological distress above the accepted GHQ12 cut-off point. However, distress was not related to variables such as low socio-economic status, as had been predicted. Early parenting stress was greater in relatively more educated and older mothers. New motherhood is likely to be stressful, even where mothers do not have postpartum depression, and so a range of supports is required. PMID- 18416405 TI - Attention deficit hyperactivity disorder: parents' and professionals' perceptions. AB - Attention deficit hyperactivity disorder (ADHD) is a prevalent childhood psychiatric condition. This study was a qualitative investigation with parents and professionals conducted in two north London boroughs, using focus groups as well as semi-structured and narrative interviews. The aim was to explore parents' and professionals' beliefs regarding the causes of ADHD and their perceptions of service provision. The sample was drawn purposively from GP practices and voluntary support groups. Professionals were recruited via professional networks. Analysis was thematic. It was found that the views of parents and professionals differed. Professionals were more likely to see ADHD as a medical condition, while parents were more likely to see ADHD in association with socio environmental causes. Delayed diagnosis, inadequate access to information and a lack of co-ordinated care are stated as some of the reasons for parental dissatisfaction with services. Professionals emphasised the need for multidisciplinary input into the management of ADHD. The implications of these findings were that parents often battled with professionals to encourage them to see their viewpoint, access to treatment was influenced by the views of parents and professionals, and noncompliance occurred when parents had different views from professionals. PMID- 18416406 TI - Targeted home visiting intervention: the impact on mother-infant relationships. AB - This paper reviews and reflects on six papers, which describe various early home visiting interventions that resulted in positive outcomes, particularly in relation to the mother-infant relationship. The papers were published in a special issue of the Infant Mental Health Journal in 2006. We provide a brief overview of each paper, highlighting the conclusions drawn across the six papers. Key learning points for health visitors in the UK are explored throughout. Suggestions are described for improvements to the way in which health visiting services in the UK are structured in order to further enhance mother-infant relationships. The overall finding is that mother-infant relationships and interactions can be improved through early home visiting interventions over a period of time, and that this prevents childhood problems later on. It is recommended that specific patient groups be targeted and offered tailored programmes of interventions that have a relevant theoretical base, by trained nursing staff who are supported by an infant mental health consultant. PMID- 18416407 TI - Education or training? PMID- 18416409 TI - Paying to work in the NHS. PMID- 18416410 TI - Collegiality. PMID- 18416411 TI - Accuracy of impressions and casts using different implant impression techniques in a multi-implant system with an internal hex connection. AB - PURPOSE: The aim of this study was to investigate the deviations of the implant positions of both impressions and casts using different impression materials and techniques. Furthermore, the existence of a correlation between the deviations of the impression and those of the cast was investigated. MATERIALS AND METHODS: A reference model was fabricated with 5 Frialit-2 implants parallel to each other. In a standardized experimental setting, 5 stone casts were produced with 5 different techniques using polyether (A) or polyvinyl siloxane (B through E). In 3 groups, a direct technique was used with a medium-viscosity material or a putty tray material in combination with a light-viscosity syringe material (A to C). In 2 groups, an indirect technique (either 1-step [group D] or 2-step [group E] was used with a putty-tray material in combination with a light-viscosity syringe material. The center-to-center distances were measured for impressions and casts in the horizontal plane using a computer-aided microscope, and the relative and absolute deviations compared to the reference model were calculated. Analysis of variance followed by the post-hoc Scheffe test (parametric data) or the Kruskal Wallis test followed by pair-wise Mann-Whitney tests (nonparametric data) were used for statistical analyses. Deviations of impressions were compared with their respective casts using paired t tests and the Pearson correlation coefficient. RESULTS: No significant differences for the relative deviations were found for impressions (-5 to -8 microm) or casts (+7 to +16 microm). Group E produced significantly higher absolute deviations for impressions (38 microm) and casts (39 microm) compared to the other groups (11 to 18 microm and 17 to 23 microm, respectively). A significant correlation between deviation of the impression and its respective cast was found for every group (r = 0.40 to 0.80) except group D. CONCLUSIONS: The distortions in the horizontal plane of the casts obtained from the impression techniques of groups A to D would probably not affect the clinical fit of implant-retained superstructures. Because of the high variation of deviations (-113 to +124 microm), the 2-step technique cannot be recommended. The method to measure both impression and cast provided a better understanding of how inaccuracies are caused. PMID- 18416412 TI - Maxillary sinus pneumatization following extractions: a radiographic study. AB - PURPOSE: The aim of this study was to identify and measure postextraction maxillary sinus pneumatization using fixed reference lines on panoramic radiographs. MATERIALS AND METHODS: One hundred fifty-two panoramic radiographs, each of a different subject, were used to measure superoinferior differences of the sinus floor position in dentate sites in comparison with contralateral edentulous sites. Fifty-eight pairs of panoramic radiographs, each pair of the same subject, were used to measure superoinferior differences of the sinus floor position in the same site before and 6 to 67 months after extraction. All measurements were performed using an interorbital line and 2 zygomatic process lines as reference lines. Statistical correlations between the amount of expansion of the sinus and the root classification, projection length, duration after extraction, type, and number of the extracted teeth were examined. RESULTS: Postextraction expansion of the sinus in an inferior direction occurred in both comparisons (2.18 +/- 2.89 mm for dentate versus contralateral sites and 1.83 +/- 2.46 mm for the same site pre- and postextraction). A considerable amount of expansion occurred after the extraction of teeth surrounded by a superiorly curving sinus floor (5.27 +/- 1.59 mm). A larger expansion was also measured after second molar extractions in comparison to first molars and in cases of extractions of 2 or more adjacent posterior teeth. DISCUSSION: Posterior maxillary tooth extraction caused an inferior expansion of the maxillary sinus in relation to fixed anatomic landmarks, thus proving the pneumatization phenomenon after tooth loss. CONCLUSIONS: Sinus pneumatization was identified after extraction of maxillary posterior teeth. The expansion of the sinus was larger following extraction of teeth enveloped by a superiorly curving sinus floor, extraction of several adjacent posterior teeth, and extraction of second molars (in comparison with first molars). If dental implant placement is planned in these cases, immediate implantation and/or immediate bone grafting should be considered to assist in preserving the 3-dimensional bony architecture of the sinus floor at the extraction site. PMID- 18416413 TI - Biomechanical response of implant systems placed in the maxillary posterior region under various conditions of angulation, bone density, and loading. AB - PURPOSE: The aim of this study was to determine the relative contribution of changes in implant system, position, bone type, and loading condition on the biomechanical response of a single-unit implant-supported restoration using nonlinear 3-dimensional finite element analysis (3D FEA). MATERIALS AND METHODS: FEA models of a single-unit (crown) restoration supported by the Frialit-2 implant and MH-6 abutment or the Straumann standard implant with the Straumann solid abutment were used. Each system was analyzed by FEA with both straight and 20-degree angled abutments. Simulated implant placement was performed in the maxillary premolar area with 3 variations in implant orientation relative to the residual ridge. Analysis of each orientation was conducted for each of 4 bone quality types described by Lekholm and Zarb, with lateral and axial loading conditions imposed. The effect of each variable was expressed as a percentage of the total sum of squares as computed using analysis of variance. RESULTS: Larger strain values were noted in cortical bone with lateral force and the Frialit-2 system. Bone strain increased with decreasing bone density and was affected primarily by bone quality. Implant stress was influenced mainly by implant position. CONCLUSIONS: Better stress/strain distribution is possible when implants are placed along the axis of loading with multiple areas of cortical contact. The Straumann solid abutment performed better as a force-transmission mechanism. PMID- 18416414 TI - Evaluation of the cylinder implant thread height and width: a 3-dimensional finite element analysis. AB - PURPOSE: To evaluate continuous and simultaneous variations of thread height and width for an experimental screw-type implant. MATERIALS AND METHODS: A finite element model of an implant with a V-shaped thread was created. The range of thread height was set at 0.20 to 0.60 mm, and the range of thread width was set at 0.10 to 0.40 mm. Forces of 100 N and 50 N were applied along the implant axis (AX) and an angle of 45 degrees in a buccolingual direction (45-degree BL), respectively. The maximum von Mises stresses in jawbone were evaluated, and the sensitivity of the stress in jawbone to the variables was also evaluated. RESULTS: Under AX load, the maximum von Mises stresses in cortical and cancellous bones increased by 4.3% and 63.0%, respectively, as thread parameters changed. Under 45-degree BL load, maximum von Mises stresses in cortical and cancellous bones increased by 19.3% and 118.0%, respectively. When thread height was from 0.34 to 0.50 mm and thread width was 0.18 to 0.30 mm, the tangent slope of the maximum von Mises stress response curve ranged from -1 to 1. The variation of the maximum von Mises stresses in jawbone was more sensitive to thread height than to thread width. CONCLUSIONS: Stress in cancellous bone is more likely to be influenced by thread parameters than stress in cortical bone. A 45-degree BL force is more likely to be influenced by thread parameters than an axial force. A thread height of 0.34 to 0.50 mm and a thread width of 0.18 to 0.30 mm is optimal from a biomechanical point of view. In the design of a screw-type implant, thread height is more important than thread width for the reduction of stress within the bone. PMID- 18416415 TI - Skeletal anchorage in orthodontics--a review of various systems in animal and human studies. AB - PURPOSE: The aim of the present investigation was to review and evaluate the current literature on skeletal bone anchorage in orthodontics with regard to success rates of the various systems. MATERIALS AND METHODS: MEDLINE, PubMed, and Cochrane searches (period January 1966 to January 2006, English language) of animal and human studies using skeletal anchorage during orthodontic treatment were scrutinized. A total of 50 relevant articles were identified which investigated various types of implants. RESULTS: Two types of anchorage systems are used in orthodontics: (1) osseointegrated dental implants, including temporary mid-palatal implants. These systems were associated with a wide variety of success rates in animal studies. In human studies, the systems were shown to be reliable, with success rates between 85% and 100% (ie, systems still functioning at the end of the orthodontic treatment). (2) Nonosseointegrated mini plates and mini-screw anchorage systems. Titanium miniplates were associated with 100% success in animals, and hardly any loss of these mini-plates (bone anchors) were lost due to infection in human studies, with success rates between 91% and 100%. Few long-term studies on nonosseointegrated mini-screws were found, but in animal studies, success rates ranged from 90% to 100%. A success rate of more than 75% in human studies is considered favorable for these orthodontic implants, which confirms the clinical applicability of this type of immediate loading anchor support in orthodontics. CONCLUSIONS: Both animal and human studies revealed that mesiodistal and intrusion movements can be reliably carried out by means of skeletal anchorage devices. A drawback is that animal studies do not reflect the real orthodontic clinical situation; thus, the outcome of these studies should be interpreted with caution. Human studies, however, show that orthodontic forces between 100 and 400 grams can be applied successfully to skeletal anchorage devices. Appropriate treatment strategies need to be confirmed by randomized prospective clinical trials. (More than 50 references). PMID- 18416416 TI - Fate of implant-retained craniofacial prostheses: life span and aftercare. AB - PURPOSE: To assess the need for surgical and prosthetic aftercare of craniofacial prostheses supported by endosseous implants. MATERIALS AND METHODS: A retrospective clinical study assessing the surgical and prosthetic aftercare from implant placement to last visit of follow-up was performed in consecutively treated patients with implant-retained craniofacial prostheses in a department of oral and maxillofacial surgery between 1988 and 2003. RESULTS: Ninety-five patients were rehabilitated with implant-retained craniofacial prostheses. Mean follow-up was 88 months (median, 79 months). Two hundred seventy implants were placed; 153 implants in the mastoid region, 99 in the orbital region, and 18 in the nasal region. The craniofacial defects were due to genetic disorders (24 patients), trauma (12 patients), and ablative tumor surgery (59 patients). In the latter group, 104 implants (33 patients) were placed in irradiated bone. Thirty implants were lost; 8 implants in nonirradiated bone (95.2% overall implant survival rate; mastoid, 95.7%; orbit, 94.1%; nose, 87.5%) and 22 implants in irradiated bone (78.8% overall implant survival rate; mastoid, 86.2%; orbit, 73.8%; nose, 90.0%). Irrespective of the craniofacial defect, on average every 1.5 to 2 years a new facial prosthesis was made, mostly for reasons because of discoloration (31.2%), problems with attachment of the acrylic resin clip carrier to the silicone (25.3%), rupture of the silicone (13.3%), or bad fit (10.9%). Severe skin reactions around implants or beneath prostheses were only observed in the orbital region. CONCLUSION: Implant-retained craniofacial prostheses are a reliable treatment option for the restoration of craniofacial defects. The need for surgical aftercare was minor, and prosthetic aftercare predominantly consisted of making new prostheses. PMID- 18416417 TI - Biological space adaptation to implant dimensions. AB - PURPOSE: Implant osseointegration has been well described, but coronal osseous healing continues to be investigated because of its impact on esthetic results and long-term maintenance. Although numerous implant diameters and designs exist, little is known about the role of these parameters on surrounding bone. Therefore, this study aimed at elucidating the influence of implant dimensions on crestal bone morphology. MATERIALS AND METHODS: Sixty Biomet/3i implants (20 standard, 20 wide, and 20 expanded platform [XP]) were randomly placed in posterior quadrants of 5 mongrel dogs. After healing, exposure of the implants to the oral cavity, and euthanasia of the animals, samples were harvested. Histomorphometric measurements were performed to determine the bone cuff height, width, and angle, and analysis of variance was applied to compare groups. RESULTS: Formation of a periimplant cuff was noticed in all implant sites. Mean cuff height was 0.8 mm, 1 mm, and 1.4 mm for standard, wide, and XP implants, respectively. Mean cuff width was 1.9 mm, 2.1 mm, and 2.8 mm for standard, wide, and XP implants, respectively. These differences were statistically significant between wide and XP implants (P = .035), as well as between standard and XP implants (P = .001). Angle did not differ significantly between implants of different platform widths. CONCLUSIONS: Craterization after placement of healing abutments and a healing period was observed around all implants. Width and height of the cuff varied significantly with implant diameter and platform design, but the angle formed with the implant did not vary significantly. PMID- 18416418 TI - Immediate loading of splinted locking-taper implants: 1-year survival estimates and risk factors for failure. AB - PURPOSE: The purpose of this study was to estimate the 1-year survival rate of immediate vertical-load splinted locking-taper implants and to identify risk factors for implant failure. MATERIALS AND METHODS: To address the research aim, the investigators implemented a retrospective cohort study design and enrolled a sample derived from the population of patients who had received immediate vertical-load splinted implants (Bicon, Boston, MA). The predictor variables were the sets of exposures associated with implant failure and classified as demographic, health status, anatomic, implant specific, prosthetic, and surgical. The primary outcome variable was implant failure, which was defined as implant removal. Descriptive, univariate, and multivariate analyses using clustered marginal approach of the Cox proportional hazards models were computed. The level of statistical significance was set at P < .05. RESULTS: The study cohort was composed of 209 patients who received 477 implants. The overall 1-year Kaplan Meier survival estimate was 90.3% (95% CI: 86.9%, 93.7%). In the multivariate model, implant placement in a delayed manner versus implantation the same day as extraction (hazard ratio = 3.7, P = .002), uncoated implants versus coated implants (hazard ratio = 22.1, P < .001), and an increased per-unit number of pontics involved in the temporary prosthesis (hazard ratio = 1.8, P < .001) were statistically associated with an increased risk of implant failure. CONCLUSIONS: An overall 1-year survival estimate of 90.3% (95% CI: 86.9%, 93.7%) was calculated for immediately loaded splinted implants. After controlling for other variables, 3 variables-timing of implant placement relative to extraction (delayed implant placement after tooth extraction), coating of implant (uncoated), and increased number of pontics-were associated with an increased risk for implant failure. PMID- 18416419 TI - GBR and autogenous cortical bone particulate by bone scraper for alveolar ridge augmentation: a 2-case report. AB - Scientific literature describes autogenous bone as the gold standard among graft materials for alveolar reconstructive procedures. Alveolar ridge augmentation has been clinically achieved with different forms of autogenous bone, including autogenous cortical bone particulate (ACBP). However, few histologic studies demonstrating the biologic potential and healing dynamics following the use of ACBP are currently available. This case report presents 2 patients in whom atrophic edentulous alveolar crests were submitted to a vertical/lateral ridge augmentation prior to implant placement. The technique was performed through the use of a titanium-reinforced expanded polytetrafluoroethylene (e-PTFE) membrane with an ACBP graft obtained from the retromolar region with a specially designed bone scraper. Bone biopsy specimens were harvested at 9 months after graft placement. Analysis of the reconstructed bone revealed bone with a lamellar quality characterized by a mature osteonic structure. Sparse particles of grafted bone were evident in direct contact with the regenerated bone. Marrow spaces showed a normal stromal component with limited grafted particles. PMID- 18416420 TI - Dental rehabilitation using an implant-carrying plate system in a severely resorbed edentulous maxilla: a case report. AB - This clinical article describes a case of dental rehabilitation using an implant carrying plate system (EPITEC) for a patient with severely resorbed edentulous maxilla and microstomia. In this case, the presence of microstomia prevented bone augmentation procedures through an intraoral approach. Treatment using 2 endosseous implants inserted in the canine regions and an implant-supported overdenture was planned. However, endosseous implants were not feasible on the right side because of insufficient available bone volume. An implant-carrying plate system was then utilized on the right side. Four months later, an implant supported ball-attachment overdenture was fabricated. At the 2-year follow-up, the clinical course remained uneventful, and the patient remained satisfied with the treatment. PMID- 18416421 TI - Ideal implant positioning in an anterior maxillary extraction socket by creating an apico-palatal guiding slot: a technical note. AB - In anterior maxillary extraction sockets, immediate implants are usually positioned along the palatal slope of the extraction socket. However, this positioning frequently causes unintentional slipping of dental implant toward thin labial plate, leading to its fracture or perforation. To prevent this, the present authors have created a simple apico-palatal guiding slot in the extraction socket following atraumatic extraction. Guided by this slot, sequential drilling up to implant placement can be easily accomplished while maintaining ideal axial orientation of the dental implant. PMID- 18416422 TI - A bovine-bone mineral block for the treatment of severe ridge deficiencies in the anterior region: a clinical case report. AB - A bovine-bone mineral block was used to treat a severe horizontal and vertical anterior ridge deficiency. Such a block can be shaped to conform to the defect, and it avoids the need for harvesting autogenous bone or fixation of the block with screws. After a 6-month integration period, an implant was placed. Six months later the implant was restored with a single crown. The case has been followed for 3 years. Slow bone resorption has been observed at the interface between the block and the surrounding bone, but the interproximal bone peaks, important for soft tissue support and esthetics, have been maintained over time. The 3-year follow-up results suggest that bovine-bone mineral blocks may be a suitable bone-replacement material for augmentation of extensive alveolar ridge defects in the anterior region. PMID- 18416423 TI - Benign paroxysmal vertigo secondary to placement of maxillary implants using the alveolar expansion technique with osteotomes: a study of 4 cases. AB - The osteotome method is an often-used technique of great utility in certain patients with maxillary bone atrophy. However, it has been associated with the provocation of benign paroxysmal positional vertigo (BPPV), which has been described as a consequence of working the implant bed with osteotomes. During the placement of maxillary dental implants using the osteotome technique, the trauma induced by percussion with the surgical hammer, along with hyperextension of the neck during the operation, can displace otoliths and induce BPPV. Four cases of BPPV occurring after the preparation of maxillary implant beds are presented. Treatment consists fundamentally of maneuvers to move the calcium carbonate crystals from their anomalous location in the semicircular canal to their correct place in the utricle. PMID- 18416424 TI - Endosseous implant placement in conjunction with inferior alveolar nerve transposition: a report of an unusual complication and surgical management. AB - Inferior alveolar nerve transposition and placement of endosseous implants is one of the treatment options for patients with an edentulous posterior mandible with inadequate bone height superior to the inferior alveolar canal. The possible complications associated with this technique include prolonged neurosensory disturbances, infection, and pathologic fracture. This report presents the surgical management of a patient who sustained a mandibular fracture after inferior alveolar nerve transposition for the placement of 3 endosseous implants. PMID- 18416425 TI - Severe osteomyelitis following immediate placement of a dental implant. AB - Risk factors and complications in immediate implant placement are widely discussed. The present report describes a case of severe osteomyelitis as a serious complication after the immediate placement of a dental implant into an extraction socket of a 61-year-old woman. The course leads from initial treatment of recurrent perimandibular abscesses with surgical drainage and high-dose intravenous antibiotics to a refractory osteomyelitis. Hemimandibulectomy and partial mandibular reconstruction with a free fibular flap followed. PMID- 18416426 TI - Closure of oroantral communication with buccal fat pad flap in zygomatic implant surgery: a case report. AB - The scientific literature has demonstrated the use of the buccal fat pad (BFP) flap to cover bone grafts in the correction of maxillary osseous defects and in the closure of oroantral communications. The use of the pedicled BFP flap to provide an immediate blood supply to a recipient site is recommended to provide closure of oroantral communications. The author presents a case report of zygomatic implant surgery in which the BFP flap technique was used in the closure of an oroantral communication caused by maxillofacial surgery. PMID- 18416427 TI - Alveolar cleft closure by distraction osteogenesis with skeletal anchorage during consolidation. AB - Intraoral distraction osteogenesis (DO) has been widely used for the reconstruction of various dentoalveolar defects. However, its use in the management of alveolar clefts is relatively new. This method allows the closure of the cleft via the regeneration of new alveolar bone and attached gingiva through the distraction of a dento-osseous segment. It eliminates the need for a donor site for autogenous bone grafting and possible graft failure. However, the relatively long consolidation period required for the use of intraoral DO devices may result in soft tissue irritation that would compromise patient cooperation, especially in children. In the case presented, the intraoral DO technique was used for the treatment of a unilateral residual alveolar cleft and an implant was subsequently placed in the regenerated bone. A miniplate was also placed to serve as a skeletal anchor to enable the early removal of the distractor device. The distractor was removed before the beginning of the consolidation phase. PMID- 18416428 TI - Peri-implant bone loss caused by occlusal overload: repair of the peri-implant defect following correction of the traumatic occlusion. A case report. AB - The purpose of this case report is to demonstrate the relation between occlusal overload and peri-implant bone loss and the reversal of the situation after removal of the offending forces. The placement of an unstable removable prosthesis on 3 well-integrated implants that had been stable for 9 years caused noticeable bone loss after 6 months. The elimination of the traumatic occlusion reversed the situation, and a remarkable healing of the peri-implant tissue occurred until the pretrauma condition was nearly restored. The condition has been stable for the past 4 years. PMID- 18416430 TI - Nanomosaic network for the detection of proteins without direct electrical contact. AB - A nanomosaic network of metallic nanoparticles for the detection of ultralow concentrations of proteins is reported, which uses two planar microelectrodes embedded in a microchip that permit generation of capacitive coupling to the nanomosaic system without the need for direct electrical contact with the channel. By tailoring the microchannel surface using a sandwich configuration of polyethylene terephthalate/gold nanoparticles/poly(L-lysine), the surface charge can be modified following biomolecular interactions and monitored using a noncontact admittance technique. This nanodevice system behaves like a tunable capacitor and can be employed for the detection of any kind of molecule. The femtomolar detection of an anionic protein, such as beta-lactoglobulin in phosphate-buffered saline medium, is taken as an example. PMID- 18416429 TI - Micro-organism-triggered release of silver nanoparticles from biodegradable oxide carriers allows preparation of self-sterilizing polymer surfaces. AB - The antimicrobial activity of silver has attracted significant research interest and contributes to an exponentially growing use of this noble metal in commodity products. In this investigation, we describe a general approach to increase the antimicrobial activity of a silver-containing surface by two to three orders of magnitude. The use of 1-2-nm silver particles decorating the surface of 20-50-nm carrier particles consisting of a phosphate-based, biodegradable ceramic allows the triggered release of silver in the presence of a growing microorganism. This effect is based on the organism's requirements for mineral uptake during growth creating a flux of calcium, phosphate, and other ions to the organism. The growing micro-organism dissolves the carrier containing these nutrients and thereby releases the silver nanoparticles. Further, we demonstrate the rapid self sterilization of polymer surfaces containing silver on calcium phosphate nanoparticles using a series of human pathogens. Colony-forming units (viable bacteria or fungi counts) have been routinely reduced below detection limit and suggest application of these self-sterilizing surfaces in hospital environments, food and pharmaceutical processing, and personal care. PMID- 18416431 TI - Characterization of the pH-dependent dissociation of a multimeric metalloprotein Streptomyces rubiginosus xylose isomerase by ESI FT-ICR mass spectrometry. AB - We report an analysis of the pH-dependent dissociation of a multimeric metalloprotein, xylose isomerase from Streptomyces rubiginosus (XI), by electrospray ionization (ESI) Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometry. Xylose isomerases are industrially significant enzymes that catalyze interconversion of aldose and ketose sugars. XI is biologically active as a approximately 173-kDa tetrameric complex, comprised of four identical approximately 43-kDa subunits and eight metal cations, unequivocally identified as the Mg(2+) cations in this work. ESI FT-ICR mass spectra of XI measured in the pH range of 3.0-6.9 indicated that the dissociation of the intact holo-tetramer is initiated by the loss of all eight Mg(2+) cations at pH O-->C7') ether linkages have been located in sequences of PAs ranging in chain lengths of two to five monomer units using ESI-MS(n) data. Using the fragmentation information from ESI-MS(n) experiments, a total of 17 PAs were structurally sequenced by systematic real time ESI-MS(n). Among them ten A-type and six B-type hop PAs are reported here for the first time. PMID- 18416439 TI - Sample size for equivalence trials: a case study from a vaccine lot consistency trial. AB - For some trials, simple but subtle assumptions can have a profound impact on the size of the trial. A case in point is a vaccine lot consistency (or equivalence) trial. Standard sample size formulas used for designing lot consistency trials rely on only one component of variation, namely, the variation in antibody titers within lots. The other component, the variation in the means of titers between lots, is assumed to be equal to zero. In reality, some amount of variation between lots, however small, will be present even under the best manufacturing practices. Using data from a published lot consistency trial, we demonstrate that when the between-lot variation is only 0.5 per cent of the total variation, the increase in the sample size is nearly 300 per cent when compared with the size assuming that the lots are identical. The increase in the sample size is so pronounced that in order to maintain power one is led to consider a less stringent criterion for demonstration of lot consistency. The appropriate sample size formula that is a function of both components of variation is provided. We also discuss the increase in the sample size due to correlated comparisons arising from three pairs of lots as a function of the between-lot variance. PMID- 18416440 TI - Measurement error correction for nutritional exposures with correlated measurement error: use of the method of triads in a longitudinal setting. AB - Nutritional exposures are often measured with considerable error in commonly used surrogate instruments such as the food frequency questionnaire (FFQ) (denoted by Q(i) for the ith subject). The error can be both systematic and random. The diet record (DR) denoted by R(i) for the ith subject is considered an alloyed gold standard. However, some authors have reported both systematic and random errors with this instrument as well.One goal in measurement error research is to estimate the regression coefficient of T(i) (true intake for the ith subject) on Q(i) denoted by lambda(TQ). If the systematic errors in Q(i) and R(i) (denoted by q(i) and r(i)) are uncorrelated, then one can obtain an unbiased estimate of lambda(TQ) by lambda(RQ) obtained by regressing R(i) on Q(i). However, if Corr(q(i), r(i))>0, then lambda(RQ)>lambda(TQ).In this paper, we propose a method for indirectly estimating lambda(TQ) even in the presence of correlated systematic error based on a longitudinal design where Q(i) (surrogate measure of dietary intake), R(i) (a reference measure of dietary intake), and M(i) (a biomarker) are available on the same subjects at 2 time points. In addition, between-person variation in mean levels of M(i) among people with the same dietary intake is also accounted for. The methodology is illustrated for dietary vitamin C intake based on longitudinal data from 323 subjects in the European Prospective Investigation of Cancer (EPIC)-Norfolk study who provided two measures of dietary vitamin C intake from the FFQ (Q(i)) and a 7-day DR (R(i)) and plasma vitamin C (M(i)) 4 years apart. PMID- 18416441 TI - Rapid screening and characterization of drug metabolites using a multiple ion monitoring-dependent MS/MS acquisition method on a hybrid triple quadrupole linear ion trap mass spectrometer. AB - A novel LC/MS/MS method that uses multiple ion monitoring (MIM) as a survey scan to trigger the acquisition of enhanced product ions (EPI) on a hybrid quadrupole linear ion trap mass spectrometer (Q TRAP) was developed for drug metabolite identification. In the MIM experiment, multiple predicted metabolite ions were monitored in both Q1 and Q3. The collision energy in Q2 was set to a low value to minimize fragmentation. Results from analyzing ritonavir metabolites in rat hepatocytes demonstrate that MIM-EPI was capable of targeting a larger number of metabolites regardless of their fragmentation and retained sensitivity and duty cycle similar to multiple reaction monitoring (MRM)-EPI. MIM-based scanning methods were shown to be particularly useful in several applications. First, MIM EPI enabled the sensitive detection and MS/MS acquisition of up to 100 predicted metabolites. Second, MIM-MRM-EPI was better than MRM-EPI in the analysis of metabolites that undergo either predictable or unpredictable fragmentation pathways. Finally, a combination of MIM-EPI and full-scan MS (EMS), as an alternative to EMS-EPI, was well suited for routine in vitro metabolite profiling. Overall, MIM-EPI significantly enhanced the metabolite identification capability of the hybrid triple quadrupole-linear ion trap LC/MS. PMID- 18416442 TI - Joint spatial survival modeling for the age at diagnosis and the vital outcome of prostate cancer. AB - Prostate cancer (PrCA) is the most common malignancy in men and a leading cause of cancer mortality among males in the United States. Large geographical variation and racial disparities exist in both the incidence of PrCA and the survival rate after diagnosis. In this population-based study, a joint spatial survival model is constructed to investigate factors that affect the age at diagnosis of PrCA and the subsequent survival. The joint model for these two time to-event outcomes is specified through parametric models for age at diagnosis and survival time conditional on diagnosis age. To account for possible correlation in these outcomes among men from the same geographical region, frailty terms are included in the survival model. Both spatially correlated and uncorrelated frailties are incorporated in each model considered. The deviance information criterion is used to select a best-fitting model within the Bayesian framework. The results from our final best-fitting model indicate that race, marital status at diagnosis, and cancer stage are significantly associated with both of the two time-to-event outcomes. No pattern emerged in the geographical distribution of age at PrCA diagnosis. In contrast, a spatially clustered pattern was observed in the geographic distribution of survival experience post diagnosis. PMID- 18416444 TI - Welcome and opening remarks 2007 CDC-ATSDR symposium on statistical methods. Preface. PMID- 18416445 TI - A method for the meta-analysis of mutually exclusive binary outcomes. AB - Meta-analyses of multiple outcomes need to take into account the within-study correlation across the different outcomes. Here we focus on the meta-analysis of dichotomous outcomes that are mutually exclusive and exhaustive. Correlations between effect sizes for mutually exclusive outcomes are negative and can be obtained from data already available. We present both fixed-effects and random effects methods that account for the negative correlations and yield correct simultaneous confidence intervals for both the marginal outcome-specific effect sizes and the relative effect sizes between outcomes. Formulae for the odds ratio, risk ratio, risk difference, and the differences in the arcsin-transformed risks are provided. An example of a meta-analysis of randomized trials of radiotherapy and mastectomy with axillary lymph node clearance versus only mastectomy with axillary clearance for early breast cancer is presented. The mutually exclusive outcomes of breast cancer deaths and deaths secondary to other causes are examined in separate meta-analyses, and also by taking the between outcome correlation into account. We argue that mutually exclusive outcomes in the meta-analyses of binary data are optimally analyzed in a multinomial setting. This may also be applicable when a meta-analysis examines only one out of several mutually exclusive outcomes. For large sample sizes and/or low event counts, the covariances between outcome-specific effect sizes are small, and either ignoring them or accounting for them would result in similar estimates for any practical purpose. However, meta-analysts should explore the robustness of the findings from individual meta-analyses when mutually exclusive outcomes are assessed. PMID- 18416446 TI - Application timing and efficacy of alternatives for the insecticidal control of Tipula paludosa Meigen (Diptera: Tipulidae), a new invasive pest of turf in the northeastern United States. AB - BACKGROUND: Two invasive crane flies, Tipula paludosa Meigen and Tipula oleracea L. (Diptera: Tipulidae), were detected in New York State for the first time in 2004. Both are damaging pests of turfgrass and other horticultural systems in North America where establishment has already been documented. To develop management recommendations for the Northeast and define opportunities for preventive (autumn) and curative (spring) control, four insecticide trials targeting T. paludosa larvae were conducted over a 2 year period. RESULTS: The most efficacious (>or=70% control in both trials) products against early instars in autumn were bifenthrin, carbaryl, chlorantraniliprole, clothianidin and trichlorfon. Results varied for azadirachtin, Beauveria bassiana, cyfluthrin, dinotefuran, imidacloprid, imidacloprid + bifenthrin and indoxacarb. Clothianidin and dinotefuran were most efficacious against fourth instars in spring; results varied for imidacloprid, indoxacarb and trichlorfon. CONCLUSION: Several insecticides offer alternatives for preventive and curative control of T. paludosa, but, because there is little overlap with application windows for scarab larvae pests, management may entail an entirely new insecticide treatment window, implying new economic and environmental burdens to the turfgrass industry. Moreover, curtailing the impact and spread of these invasives may be severely hampered because the best performing alternatives (clothianidin, dinotefuran) are not registered in New York. PMID- 18416447 TI - A two-level structural equation model approach for analyzing multivariate longitudinal responses. AB - The analysis of longitudinal data to study changes in variables measured repeatedly over time has received considerable attention in many fields. This paper proposes a two-level structural equation model for analyzing multivariate longitudinal responses that are mixed continuous and ordered categorical variables. The first-level model is defined for measures taken at each time point nested within individuals for investigating their characteristics that are changed with time. The second level is defined for individuals to assess their characteristics that are invariant with time. The proposed model accommodates fixed covariates, nonlinear terms of the latent variables, and missing data. A maximum likelihood (ML) approach is developed for the estimation of parameters and model comparison. Results of a simulation study indicate that the performance of the ML estimation is satisfactory. The proposed methodology is applied to a longitudinal study concerning cocaine use. PMID- 18416448 TI - Consultants' forum: should post hoc sample size calculations be done? AB - Pre-study sample size calculations for clinical trial research protocols are now mandatory. When an investigator is designing a study to compare the outcomes of an intervention, an essential step is the calculation of sample sizes that will allow a reasonable chance (power) of detecting a pre-determined difference (effect size) in the outcome variable, at a given level of statistical significance. Frequently studies will recruit fewer patients than the initial pre study sample size calculation suggested. Investigators are faced with the fact that their study may be inadequately powered to detect the pre-specified treatment effect and the statistical analysis of the collected outcome data may or may not report a statistically significant result. If the data produces a "non statistically significant result" then investigators are frequently tempted to ask the question "Given the actual final study size, what is the power of the study, now, to detect a treatment effect or difference?" The aim of this article is to debate whether or not it is desirable to answer this question and to undertake a power calculation, after the data have been collected and analysed. PMID- 18416449 TI - Effect of genotype of cocoa (Theobroma cacao) on attractiveness to the mirid Sahlbergella singularis (Hemiptera: Miridae) in the laboratory. AB - BACKGROUND: Mirids are a major constraint to cocoa growing in Africa. Cocoa breeding for tolerance/resistance could be effective in an integrated pest management system. Attractiveness is one aspect of tolerance/resistance, and decreasing the attractiveness of cocoa trees should be a good way of reducing damage. RESULTS: Small-scale laboratory tests were carried out in Cameroon to assess differences in the attractiveness to mirids of eleven cocoa genotypes. The genotypes were ranked according to their attractiveness score and a distance from a mean value. An analysis of variance was performed and revealed significant differences between cocoa genotypes (F=3.15, P<0.001). The cocoa genotype groupings revealed three major categories, with BE10 and SNK413 proving to be the most attractive. In contrast, five genotypes, IMC60, the Catongo Trinitario genotypes, Playa Alta2 (from Venezuela), SIC5 and SNK614, proved to be less attractive than the mean. Four genotypes (PA107, SNK619, UPA134 and T60/887) displayed similar attractiveness to the mean. CONCLUSION: The circular microtest offered the advantage of not needing a reference cocoa genotype. The least attractive clones, such as IMC60, were also the most tolerant in the field. Comparisons with the results of other studies are proposed. PMID- 18416450 TI - Anti-HIV adaptive immunity: determinants for viral persistence. AB - The immense difficulty in primary control of human immunodeficiency virus (HIV) and simian immunodeficiency virus (SIV) infection by adaptive immune responses has been a topic of exceptional importance. CD8+ cytotoxic T lymphocytes (CTLs) do play a central role in primary resolution of viremia, but their potency in viral control is generally constrained in the natural courses of HIV/SIV infections. The overall repertoire of CTLs is dependent on both the host and the virus genetic polymorphisms, and the potency of each individual CTL is affected by immunological and virological determinants. HIV/SIV infections lack early appearance of neutralising antibodies (NAbs), and our recent finding has suggested a possibility of their absence contributing to diminished virus specific CD4+ T-cell responses leading to failure in primary viral control. Extrapolations from studies in macaque models of SIV infection and analyses of the cohorts of HIV control in humans have to date delineated the numerous requirements for attainment of viral control. Understanding of the individual components of adaptive immune responses and their optimal concert required for HIV/SIV control would contribute to development of an effective AIDS vaccine. Here, we discuss current insights into CTLs and NAbs, and speculate their possible protective mechanism against establishment of persistent HIV/SIV infection. PMID- 18416451 TI - Direct costs associated with mild cognitive impairment in primary care. AB - BACKGROUND/AIMS: Little is known about the direct costs of individuals with Mild Cognitive Impairment (MCI). This study investigates the direct costs associated with MCI according to recent diagnostic criteria from a societal perspective. METHODS: Four hundred and fifty-two primary care patients aged 75+ from Leipzig, Germany, were investigated in face-to-face interviews regarding MCI according to the current diagnostic criteria of the International Working Group on MCI, resource utilisation and costs (questionnaire of service utilisation and costs), as well as chronic medical illness (Chronic Disease Score). Resource utilisation was monetarily valued using 2004/2005 prices. RESULTS: Mean annual direct costs were 4,443 euro for patients with MCI (n=39) and 3,814 euro for patients without MCI (n=413) (p=0.34). Looking at the cost components, patients with and without MCI only significantly differed regarding pharmaceutical costs (1,210 euro vs 1,062 euro; p<0.05) not caused by antidementive drugs. CONCLUSION: Direct costs of individuals having MCI are not significantly increased in comparison to direct costs of individuals without cognitive deficits. PMID- 18416452 TI - Clustering and switching in semantic fluency: predictors of the development of Alzheimer's disease. AB - OBJECTIVE: The aims of the study are twofold: (1) to compare semantic fluency, clustering and switching performance among subjects with memory complaints, patients with Alzheimer Disease (AD), and healthy controls; and (2) to examine the clinical utility of the clustering/switching scoring system in the prediction of incident AD in subjects with memory complaints. METHODS: A semantic fluency task was used to compare thirty eight subjects with memory complaints, forty two AD patients and twenty five healthy controls on the total number of words generated, clustering and switching performance. Subjects with memory complaints were followed-up for a maximum period of two years and re-evaluated. They remained in the memory complaints group (twenty eight subjects) or were defined as probable AD (ten subjects). RESULTS: AD patients generated fewer correct words (p < 0.001) and showed a reduction in clustering (p = 0.008) and switching (p < 0.001). Subjects with memory complaints showed a significant reduction in correct words (p < 0.001) and clustering performance (p = 0.008) compare to controls. In the first evaluation, the subgroup of patients who converted to AD at follow up produced less correct words (p < 0.01) and smaller clusters (p = 0.007) than the subgroup who did not become demented. There were no differences in switching between these two subgroups. AD development was better predicted by cluster size than by the total number of words generated or by switching. CONCLUSIONS: Subjects with memory complaints and AD patients have an alteration in both qualitative and quantitative aspects of semantic fluency. A clustering analysis could enhance the reliability of early AD diagnosis. PMID- 18416453 TI - Opisthorchiasis in Thailand: review and current status. AB - Opisthorchiasis caused by Opisthorchis viverrini (O. viverrini) remains a major public health problem in many parts of Southeast Asia including Thailand, Lao PDR, Vietnam and Cambodia. The infection is associated with a number of hepatobiliary diseases, including cholangitis, obstructive jaundice, hepatomegaly, cholecystitis, chole-lithiasis and cholangiocarcinoma. The liver fluke infection was induced by eating raw or uncooked fish products that is the tradition and popular in the northeastern and northern region, particularly in rural areas of Thailand. Health education programs to prevent and control opisthorchiasis are still required in high-risk areas. PMID- 18416454 TI - Gene therapy: regulations, ethics and its practicalities in liver disease. AB - Gene therapy is a new and promising approach which opens a new door to the treatment of human diseases. By direct transfer of genetic materials to the target cells, it could exert functions on the level of genes and molecules. It is hoped to be widely used in the treatment of liver disease, especially hepatic tumors by using different vectors encoding the aim gene for anti-tumor activity by activating primary and adaptive immunity, inhibiting oncogene and angiogenesis. Despite the huge curative potential shown in animal models and some pilot clinical trials, gene therapy has been under fierce discussion since its birth in academia and the public domain because of its unexpected side effects and ethical problems. There are other challenges arising from the technique itself like vector design, administration route test and standard protocol exploration. How well we respond will decide the fate of gene therapy clinical medical practice. PMID- 18416455 TI - Inhibition of CXCR4 activity with AMD3100 decreases invasion of human colorectal cancer cells in vitro. AB - AIM: To investigate the effect and mechanism of blockade of the CXC chemokine receptor-4 (CXCR4) signaling pathway by AMD3100, a small non-peptide CXCR4 inhibitor, on invasion and metastasis of colorectal cancer cells in vitro. METHODS: Human colorectal cancer cell line SW480 was treated with AMD3100 at different final concentrations. 3-(4,5-dimethylthiazole-2-yl)-2.5-dipheny ltetrazolium bromide (MTT) assay was used to detect the effect of AMD3100 on cell proliferation. The invasion ability of SW480 cells was determined by cell invasion assay kit. In the presence of AMD3100, the CXCL12-mediated migratory response of SW480 cells was tested by classical chemotaxis assays. RT-PCR analysis and Western blotting were used to detect the expression of vascular endothelial growth factor (VEGF), matrix metalloproteinase-2 (MMP-2) and -9 (MMP 9) in SW480 cells. RESULTS: Cell viability was significantly suppressed by AMD3100 in a dose-dependent manner. AMD3100 (100 and 1000 ng/mL) significantly inhibited the invasion ability of SW480 cells. Treatment with AMD3100 markedly reduced the expression of VEGF and MMP-9 but not MMP-2 in SW480 cells. CONCLUSION: The CXCL12/CXCR4 system is an important mediator of proliferation and invasion of CXCR4-expressing colorectal cancer cells. AMD3100 inhibited invasion and metastasis activity of the colorectal cancer cell line SW480 through down regulation of VEGF and MMP-9 expression. PMID- 18416456 TI - Munc18/SNARE proteins' regulation of exocytosis in guinea pig duodenal Brunner's gland acini. AB - AIM: To examine the molecular mechanism of exocytosis in the Brunner's gland acinar cell. METHODS: We used a submucosal preparation of guinea pig duodenal Brunner's gland acini to visualize the dilation of the ductal lumen in response to cholinergic stimulus. We correlated this to electron microscopy to determine the extent of exocytosis of the mucin-filled vesicles. We then examined the behavior of SNARE and interacting Munc18 proteins by confocal microscopy. RESULTS: One and 6 micromol/L carbachol evoked a dose-dependent dilation of Brunner's gland acini lumen, which correlated to the massive exocytosis of mucin. Munc18c and its cognate SNARE proteins Syntaxin-4 and SNAP-23 were localized to the apical plasma membrane, and upon cholinergic stimulation, Munc18c was displaced into the cytosol leaving Syntaxin-4 and SNAP-23 intact. CONCLUSION: Physiologic cholinergic stimulation induces Munc18c displacement from the Brunner's gland acinar apical plasma membrane, which enables apical membrane Syntaxin-4 and SNAP-23 to form a SNARE complex with mucin-filled vesicle SNARE proteins to affect exocytosis. PMID- 18416457 TI - Myelophil, a mixture of Astragali Radix and Salviae Radix extract, moderates toxic side effects of fluorouracil in mice. AB - AIM: To evaluate the efficacy of Myelophil, an extract containing Astragali Radix and Salviae Radix, for reducing complications induced by 5-fluorouracil (5-FU) in a gastrointestinal cancer model. METHODS: We injected 5-FU into mice and then administered Myelophil to examine the ability of the drug to treat the side effects of 5-FU in mice. Peripheral blood counts, histological examinations, and colony-forming assays of bone marrow were conducted, followed by swimming tests and assessment of survival times. RESULTS: Myelophil restored red and white blood cells and platelets in blood, and recovered cell density in bone marrow to levels comparable to those observed within the control group. In addition, Myelophil significantly increased colony-forming unit granulocyte-macrophage (CFU-GM) and CFU-erythroid (CFU-E) compared to the control group. We confirmed that interleukin-3 gene expression was upregulated by Myelophil in spleen cells. Myelophil administration also doubled the survival rate of mice that were severely myelosuppressed as a result of 5-FU injection at a lethal dose of 70%. Finally, the swimming performance of mice significantly improved as a result of Myelophil treatment. CONCLUSION: These results provide experimental evidence in support of clinical applications of Myelophil to minimize 5-FU-induced myelosuppression and improve general post-chemotherapy health. PMID- 18416458 TI - Proliferation of L02 human hepatocytes in tolerized genetically immunocompetent rats. AB - AIM: To investigate whether human hepatocytes could proliferate after transplantation to normal immunocompetent rats treated with 2-acetaminofluorene or retrorsine and partial hepatectomy. METHODS: L02 hepatocyte-tolerant Sprague Dawley rats were injected with retrorsine, 2-acetaminofluorene or normal saline. L02 hepatocytes were then transplanted via the spleen. Human albumin and its mRNA, specific proliferating cell nuclear antigen (PCNA), L02 hepatocyte dynamic distribution, number density and area density of PCNA-positive cells in the liver were determined. RESULTS: All the examined indicators were not significantly different between the rats treated with 2-acetaminofluorene and normal saline, which was not the case with rats treated with retrorsine. A dynamic distribution of L02 hepatocytes in the rat liver was detected from wk 1 to mo 6 after transplantation in the retrorsine group and from wk 1 to 10 in the 2 acetaminofluorene group. Human albumin and its mRNA were detected from wk 2 to mo 6 in the retrorsine group and from wk 1 to 8 in the 2-acetaminofluorene group. Specific human PCNA was detected in the rat liver from wk 2 to mo 6 in the retrorsine group and from wk 2 to 6 in the 2-acetaminofluorene group. Human albumin and its mRNA contents as well as the number of PCNA positive cells reached a peak at wk 4. CONCLUSION: L02 human hepatocytes could not proliferate significiantly after transplantation to the normal, immunocompetent rats treated with 2-acetaminofluorene. L02 human hepatocytes can survive for 10 wk after transplantation and express human albumin for 8 wk. L02 human hepatocytes can proliferate and express human albumin for 6 mo after transplantation to the rats treated with retrorsine. The chimeric L02 human hepatocytes, which then underwent transplantation into tolerant rats, were normal in morphogenesis, biochemistry and function. PMID- 18416459 TI - Preservation of non-heart-beating donor livers in extracorporeal liver perfusion and histidine-trytophan-ketoglutarate solution. AB - AIM: To compare the preservation of non-heart-beating donor (NHBD) livers in cold histidine-trytophan-ketoglutarate (HTK) solution and extracorporeal liver perfusion (ECLP). METHODS: Livers harvested from healthy pigs were stored for 10 h in cold HTK solution (group A, n = 4) or perfused with oxygenated autologous blood at body temperature (group B, n = 4). Both groups were then tested on the circuit for 4 h. Bile production, hemodynamic parameters, hepatocyte markers and reperfusion injury of extracorporeal livers were tested in each group. Liver tissues from each group were examined at the end of reperfusion. RESULTS: At 1, 2, 3 and 4 h after reperfusion, bile production, hemodynamic parameters, hepatocyte markers and reperfusion injury of livers in group A were statistically different from those in group B (P < 0.05 or P < 0.01). CONCLUSION: ECLP is better than HTK solution to preserve NHBD livers. ECLP can assess the graft viability before liver transplantation. PMID- 18416460 TI - Effect of Chaiqinchengqi decoction on sarco/endoplasmic reticulum Ca2+-ATPase mRNA expression of pancreatic tissues in acute pancreatitis rats. AB - AIM: To investigate the effect of Chaiqinchengqi decoction (CQCQD) on sarco/endoplasmic reticulum Ca2+-ATPase (SERCA) mRNA expression of pancreatic tissues in acute pancreatitis (AP) rats. METHODS: Thirty Sprague-Dawley (SD) rats were randomized into control group, AP group and CQCQD group (n = 3 x 10). The rats in the CQCQD group were intragastrically administered with CQCQD (10 mL/kg every 2 h) after induction of AP by intraperitoneal injection of caerulein (50 microg/kg.h x 5) within 4 h. At 6 h after the induction of AP model, pancreatic tissues were collected for the pathological observation, mRNA extraction for determination of SERCA1 and SERCA2 mRNA expression or pancreatic acinar cell isolation for measurement of fluorescence intensity (FI) of intracellular calcium ion concentration [Ca2+]i. RESULTS: There was no expression of pancreatic SERCA1 mRNA in the control group and the AP group. The expression of pancreatic SERCA2 mRNA in the AP group was down-regulated (expression ratio = 0.536; P = 0.001) compared with the control group, while that in the CQCQD group was up-regulated (expression ratio = 2.00; P = 0.012) compared with AP group. The FI of intracellular [Ca2+] of pancreatic acinar cells in the AP group (138.2 +/- 23.1) was higher than the C group (111.0 +/- 18.4) and the CQCQD group (118.7 +/- 15.2 ) (P < 0.05) and the pancreatic pathological score in the CQCQD group was lower than that in the AP group (5.7 +/- 1.9 vs 9.2 +/- 2.7, P < 0.05). CONCLUSION: CQCQD can up-regulate the expression of SERCA2 mRNA of pancreatic tissues, reduce intracellular calcium overload and relieve pancreatic tissue lesions. PMID- 18416462 TI - Effects of alpha-adrenoreceptor antagonists on apoptosis and proliferation of pancreatic cancer cells in vitro. AB - AIM: To discuss the expression of alpha-adrenoreceptors in pancreatic cancer cell lines PC-2 and PC-3 and the effects of alpha1- and alpha2-adrenoreceptor antagonists, yohimbine and urapidil hydrochloride, on the cell lines in vitro. METHODS: We cultured the human ductal pancreatic adenocarcinoma cell lines PC-2 and PC-3 and analyzed the mRNA expression of alpha1- and alpha2-adrenergic receptors by reverse transcription polymerase chain reaction (RT-PCR). The effects of yohimbine and urapidil hydrochloride on cell proliferation were assessed by 3-(4,5-dimethylthiasol-2-yl)-2,4,-diphenyltetrazolium bromide (MTT) assay. Apoptosis was detected using the terminal deoxyribonucleotidyl transferase (TdT)-mediated biotin-16-dUTP nick-end labeling (TUNEL) assay and flow cytometry (FCM). RESULTS: PC-2 expressed mRNA in alpha1- and alpha2-adrenoreceptors. MTT assays showed that urapidil hydrochloride had no effect on PC-3 cell lines. However, exposure to urapidil hydrochloride increased DNA synthesis in PC-2 cell lines as compared to the control group. PC-2 cell lines were sensitive to both drugs. The proliferation of the 2 cell lines was inhibited by yohimbine. Cell proliferation was inhibited by yohimbine via apoptosis induction. CONCLUSION: The expression of alpha1- and alpha2-adrenoreceptors is different in PC-2 and PC-3 cell lines, which might be indicative of their different functions. The alpha2 adrenoceptor antagonist, yohimbine, can inhibit the proliferation of both cell lines and induce their apoptosis, suggesting that yohimbine can be used as an anticancer drug for apoptosis of PC-2 and PC-3 cells. PMID- 18416461 TI - Vascular endothelial growth factor attenuates hepatic sinusoidal capillarization in thioacetamide-induced cirrhotic rats. AB - AIM: To investigate the effect of vascular endothelial growth factor (VEGF) transfection on hepatic sinusoidal capillarization. METHODS: Enhanced green fluorescent protein (EGFP)/VEGF transfection was confirmed by immunofluorescence microscopy and immunohistochemistry both in primary hepatocytes and in normal liver. Cirrhotic rats were generated by thioacetamide (TAA) administration and then divided into a treatment group, which received injections of 400 microg of plasmid DNA encoding an EGFP-VEGF fusion protein, and a blank group, which received an equal amount of normal saline through the portal vein. The portal vein pressure was measured in the normal and cirrhotic state, in treated and blank groups. The average number of fenestrae per hepatic sinusoid was determined using transmission electron microscopy (TEM), while the relative abundance of VEGF transcripts was examined by Gene array. RESULTS: Green fluorescent protein was observed in the cytoplasms of liver cells under immunofluorescence microscopy 24 h after transfection with EGFP/VEGF plasmid in vitro. Staining with polyclonal antibodies against VEGF illustrated that hepatocytes expressed immunodetectable VEGF both in vitro and in vitro. There were significant differences in the number of fenestrae and portal vein pressures between normal and cirrhotic rats (7.40 +/ 1.71 vs 2.30 +/- 1.16 and 9.32 +/- 0.85 cmH2O vs 17.92 +/- 0.90 cmH2O, P < 0.01), between cirrhotic and treated rats (2.30 +/- 1.16 cmH2O vs 4.60 +/- 1.65 and 17.92 +/- 0.90 cmH2O vs 15.52 +/- 0.93 cmH2O, P < 0.05) and between the treatment group and the blank group (4.60 +/- 1.65 cmH2O vs 2.10 +/- 1.10 cmH2O and 15.52 +/- 0.93 cmH2O vs 17.26 +/- 1.80 cmH2O, P < 0.05). Gene-array analysis revealed that the relative abundance of transcripts of VEGF family members decreased in the cirrhotic state and increased after transfection. CONCLUSION: Injection of a plasmid encoding VEGF through the portal vein is an effective method to induce the formation of fenestrae and decrease portal vein pressure in cirrhotic rats. Therefore, it may be a good choice for treating hepatic cirrhosis and portal hypertension. PMID- 18416463 TI - Efficacy, risk factors and complications of endoscopic polypectomy: ten year experience at a single center. AB - AIM: To examine the efficacy and complications of colonoscopic resection of colorectal polypoid lesions. METHODS: We retrospectively reviewed 1354 polypectomies performed on 1038 patients over a ten-year period. One hundred and sixty of these were performed for large polyps, those measuring > or = 20 mm. Size, shape, location, histology, the technique of polypectomy used, complications, drugs assumption and associated intestinal or extra intestinal diseases were analyzed. For statistical analysis, the Pearson chi2 test, NPC test and a Binary Logistic Regression were used. RESULTS: The mean patient age was 65.9 +/- 12.4 years, with 671 men and 367 women. The mean size of polyps removed was 9.45 +/- 9.56 mm while the size of large polyps was 31.5 +/- 10.8 mm. There were 388 pedunculated and 966 sessile polyps and the most common location was the sigmoid colon (41.3%). The most frequent histology was tubular adenoma (55.9%) while for the large polyps was villous (92/160 -57.5%). Coexistent malignancy was observed in 28 polyps (2.1%) and of these, 20 were large polyps. There were 17 procedural bleeding (1.3%) and one perforation. The statistical analysis showed that cancer is correlated to polyp size (P < 0.0001); sessile shape (P < 0.0001) and bleeding are correlated to cardiac disease (P = 0.034), tubular adenoma (P = 0.016) and polyp size. CONCLUSION: The endoscopic resection is a simple and safe procedure for removing colon rectal neoplastic lesions and should be considered the treatment of choice for large colorectal polyps. The polyp size is an important risk factor for malignancy and for bleeding. PMID- 18416465 TI - Changes in count and function of splenic lymphocytes from patients with portal hypertension. AB - AIM: To investigate changes in numbers and proliferative function of splenic lymphocytes in patients with hypersplenism due to portal hypertension (PH), to provide evidence for further study of immune status of the spleen during PH. METHODS: Twelve spleens from patients with hypersplenism due to PH served as the PH group, and four spleens from cases of traumatic spleen rupture were regarded as the control group. After weighing the spleen, lymphocytes were separated and counted using a cell counting plate to calculate the lymphocyte count per gram of spleen tissue (relative quantity) and total lymphocyte count in whole spleen (absolute quantity). The immunohistochemical SP method was used to observe the density and distribution of lymphocytes in the spleen. The MTT method was used to observe changes in lymphocyte proliferative function. RESULTS: As compared to the control group, the splenic lymphocytes in the PH group showed that: (1) There was no difference in distribution but a significant decrease in density; (2) the number of lymphocytes per gram of spleen (relative quantity) decreased significantly [(0.822 +/- 0.157) x 10(8) vs (1.174 +/- 0.254) x 10(8), P < 0.01]; (3) with the significant increase in the weight of the PH spleen (832.6 +/- 278.2 g vs 211.7 +/- 85.6 g, P < 0.01), the total quantity of lymphocytes (absolute quantity) increased significantly [(0.685 +/- 0.072) x 10(11) vs (0.366 +/- 0.057) x 10(11), P < 0.01]; and (4) the proliferative function of lymphocytes was enhanced: T lymphocytes, (0.022 +/- 0.005 vs 0.015 +/- 0.003, P < 0.05), and B lymphocytes (0.034 +/- 0.006 vs 0.023 +/- 0.001, P < 0.01). CONCLUSION: Although lymphocyte density in the spleen decreased in patients with PH, the total quantity of lymphocytes increased because spleen weight increased greatly, along with the proliferating function. With respect to changes in lymphocytes, PH spleens may still have immune function, although it may be disordered. However, complete evaluation of the immune function of the spleen in PH requires more research. PMID- 18416464 TI - Intraoperative ultrasound as an educational guide for laparoscopic biliary surgery. AB - AIM: To analyze the efficacy of routine intraoperative ultrasound (IOUS) as a guide for understanding biliary tract anatomy, to avoid bile duct injury (BDI) after laparoscopic cholecystectomy (LC), as well as any burden during the learning period. METHODS: A retrospective analysis was performed using 644 consecutive patients who underwent LC from 1991 to 2006. An educational program with the use of IOUS as an operative guide has been used in 276 cases since 1998. RESULTS: IOUS was highly feasible even in patients with high-grade cholecystitis. No BDI was observed after the introduction of the educational program, despite 72% of operations being performed by inexperienced surgeons. Incidences of other morbidity, mortality, and late complications were comparable before and after the introduction of routine IOUS. However, the operation time was significantly extended after the educational program began (P < 0.001), and the grade of laparoscopic cholecystitis (P = 0.002), use of IOUS (P = 0.01), and the experience of the surgeons (P = 0.05) were significant factors for extending the length of operation. CONCLUSION: IOUS during LC was found to be a highly feasible modality, which provided accurate, real-time information about the biliary structures. The educational program using IOUS is expected to minimize the incidence of BDI following LC, especially when performed by less-skilled surgeons. PMID- 18416466 TI - Analysis of the human Atox 1 homologue in Wilson patients. AB - AIM: To analyze the metallochaperone antioxidant-1 (Atox1) gene sequence in Wilson disease patients. METHODS: Mutation analysis of the four exons of the Atox1 gene including the intron- exon boundaries was performed in 63 Wilson disease patients by direct sequencing. RESULTS: From 63 selected patients no mutations were identified after the entire coding region including the intron- exon boundaries of Atox1 were sequenced. One known polymorphism within the Atox1 gene (5'UTR -99 T>C) in 31 (49%) of the Wilson patients as well as one previously undescribed variation (5'UTR -68 C>T) in 2 of the Wilson patients could be detected. Statistical analyses revealed that the existence of a variation within the Atox1- gene showed a tendency towards an earlier onset of the disease. CONCLUSION: Based on the data of this study, no major role can be attributed to Atox1 in the pathophysiology or clinical variation of Wilson disease. PMID- 18416467 TI - Primary gastric mucosa associated lymphoid tissue lymphoma: clinical data predicted treatment outcome. AB - AIM: To determine clinical characteristics and treatment outcome of gastric lymphoma after chemotherapy and immuno-chemotherapy. METHODS: Thirty four patients with primary gastric mucosa associated lymphoid tissue (MALT) lymphoma (Ann Arbor stages I to IV) were enrolled. All had upper gastric endoscopy, abdominal ultrasonography, CT and H pylori status assessment (histology and serology). After anti-H pylori treatment and initial chemotherapy, patients were re-examined every 4 mo. RESULTS: Histological regression of the lymphoma was complete in 22/34 (64.7%) and partial in 9 (26.5%) patients. Median follow up time for these 31 responders was 60 mo (range 48-120). No regression was noted in 3 patients. Among the 25 (73.5%) H pylori positive patients, the eradication rate was 100%. CONCLUSION: Using univariate analysis, predictive factors for overall survival were international prognostic index (IPI) score, hemoglobin level, erythrocyte sedimentation rate (ESR), and platelet numbers (P < 0.005). In addition to this, Cox proportion hazard model differentiate IPI score, ESR, and platelets as predictors of survival. PMID- 18416468 TI - Radiotherapy for 65 patients with advanced unresectable hepatocellular carcinoma. AB - AIM: To evaluate the efficacy of radiotherapy (RT) in patients with advanced unresectable hepatocellular carcinoma (HCC). METHODS: A total of 65 patients were treated with RT in the Korea University Medical Center. The median age of the patients was 60 years, and 86.2% were men. 18.5% and 81.5% of the patients were diagnosed as TNM stage II and IV-A, respectively. Treatment response was assessed 4 mo after initiation of RT. Tumor regression rate 1 mo after initiation of RT (TRR1m) was also assessed. Duration of survival was calculated from the initiation of RT. RESULTS: The objective treatment response was 56.9%. The 12 mo survival rate was 34.7%. Predictive factors for survival were Child-Pugh grade, alpha-fetoprotein level and treatment response. An objective response was achieved more frequently in patients with TRR1m > or = 20% than in those with TRR1m < 20% (P < 0.001). CONCLUSION: RT is effective in treating advanced HCC with a tumor response rate of 56.9%. PMID- 18416469 TI - Is there correlation between pancreatic enzyme and radiological severity in acute pancreatitis? AB - AIM: To investigate the correlation between the changes of pancreatic enzyme, the biochemical markers and the clinical results according to the Balthazar computer tomography (CT) grade. METHODS: Between July 2004 and July 2005, we reviewed the charts of 119 patients who were admitted to our hospital with acute pancreatitis. RESULTS: Eighty-three patients (69.7%) were male, and the mean age of the patients was 57 +/- 15.7 years. The biliary pancreatitis patients had an older mean age. Forty-nine patients (41.1%) had biliary pancreatitis and forty-six (38.6%) had alcoholic pancreatitis. Group 3 patients had a longer duration of pain (2.51 +/- 1.16 vs 3.17 +/- 1.30 vs 6.56 +/- 6.13, P < 0.001), a longer period of fasting (7.49 +/- 4.65 vs 10.65 +/- 5.54 vs 21.88 +/- 13.81, P < 0.001) and a longer hospital stay (9.17 +/- 5.34 vs 14.63 +/- 8.65 vs 24.47 +/- 15.52, P < 0.001) than the other groups. On the univariate analysis, the factors that affected the radiological grade were the leukocyte count at admission (P = 0.048), the hemoglobin (P = 0.016) and total bilirubin concentrations (P = 0.023), serum lipase (P = 0.009), the APACH II scores at admission (P = 0.017), the APACH II scores after 24 h (P = 0.031), the C-reactive protein (CRP) titer (P = 0.0001) and the follow up CRP titer (P = 0.003). But the CRP level (P = 0.001) and follow up CRP titer (P = 0.004) were only correlated with the radiological grade on multivariate analysis. According to the ROC curve, when we set the CRP cut off value at 83 mg/L, the likelihood ratio for a positive test was 3.84 and the likelihood ratio for a negative test was 0.26 in group 3. CONCLUSION: In conclusion, our study suggests that the CRP with the radiological severity may be used to estimate the severity of acute pancreatitis. PMID- 18416471 TI - Association between colonic polyps and diverticular disease. AB - AIM: To evaluate the association between colonic polyps and diverticular disease in Japan. METHODS: We retrospectively reviewed the medical records of 672 consecutive patients who underwent total colonoscopy between August 2006 and April 2007 at Nishinjo Hospital, Okinawa, Japan. Patients with a history of any of the following were excluded from the study: previous polypectomy, colonic resection, and inflammatory bowel diseases. The association between colonic polyps and diverticular disease was analyzed by logistic regression analysis, adjusted for age and sex. RESULTS: Prevalence of colonic polyps in all patients with diverticular disease was significantly higher than that in those without diverticular disease (adjusted odds ratio 1.7). CONCLUSION: Our data showed that patients with diverticular disease have a higher risk of colonic polyps compared to those without. PMID- 18416470 TI - Stronger inhibition of gastric acid secretion by lafutidine, a novel H2 receptor antagonist, than by the proton pump inhibitor lansoprazole. AB - AIM: To compare the antisecretory activity and plasma drug concentrations of a single oral dose of 10 mg lafutidine, a novel H2 receptor antagonist, with those of the proton pump inhibitor lansoprazole (LPZ) 30 mg. METHODS: Ten volunteers without H pylori infection participated in this crossover study comparing lafutidine 10 mg with LPZ 30 mg. Intragastric pH was monitored for 6 h in all participants, and blood samples were collected from four randomly selected individuals after single-dose administration of each drug. RESULTS: The median intragastric pH was significantly higher in individuals who received lafutidine 10 mg than in those who received LPZ 30 mg 2, 3, 4, 5, and 6 h after administration. Maximal plasma drug concentration was reached more promptly with lafutidine 10 mg than with LPZ 30 mg. CONCLUSION: In H pylori-negative individuals, gastric acid secretion is more markedly inhibited by lafutidine than by LPZ. PMID- 18416472 TI - Comparison of immediate surgical outcomes between posterior pelvic exenteration and standard resection for primary rectal cancer: a matched case-control study. AB - AIM: To determine the immediate surgical outcome and recovery of bowel function following posterior pelvic exenteration (PPE) for primary rectal cancer with suspected local invasion to the female internal reproductive organs, in comparison with a case-control series of standard resection for primary rectal cancer. METHODS: We analyzed 10 consecutive female patients undergoing PPE for the aforementioned indication between December 2003 and May 2006 in a single institution. Data were prospectively collected during hospitalization, including patient demographics, tumor- and operation-related variables and early surgical outcomes. These patients were compared with a group of female patients, matched for age, co-morbidity and location of tumor, who underwent standard resection for primary rectal cancer in the same period (non PPE group). RESULTS: In the PPE group, pathological reports showed direct invasion of the reproductive organs in 4 cases and an involvement of lymph nodes in 7 cases. A sphincter-saving operation was performed in each case. Operative time was longer (274 min vs 157 min, P < 0.001) and blood loss was greater (769 mL vs 203 mL, P = 0.008) in the PPE group. Time to first bowel movement, time to first defecation, time to resumption of normal diet, and hospital stay were not significantly different between the two groups. Postoperative complication rates were also similar. CONCLUSION: PPE for rectal cancer was associated with longer operative time and increased blood loss, but did not compromise immediate surgical outcomes and postoperative bowel function compared to standard rectal resection. PMID- 18416473 TI - Alcohol consumption and metabolic syndrome among Shanghai adults: a randomized multistage stratified cluster sampling investigation. AB - AIM: To examine the relations of alcohol consumption to the prevalence of metabolic syndrome in Shanghai adults. METHODS: We performed a cross-sectional analysis of data from the randomized multistage stratified cluster sampling of Shanghai adults, who were evaluated for alcohol consumption and each component of metabolic syndrome, using the adapted U.S. National Cholesterol Education Program criteria. Current alcohol consumption was defined as more than once of alcohol drinking per month. RESULTS: The study population consisted of 3953 participants (1524 men) with a mean age of 54.3 +/- 12.1 years. Among them, 448 subjects (11.3%) were current alcohol drinkers, including 405 males and 43 females. After adjustment for age and sex, the prevalence of current alcohol drinking and metabolic syndrome in the general population of Shanghai was 13.0% and 15.3%, respectively. Compared with nondrinkers, the prevalence of hypertriglyceridemia and hypertension was higher while the prevalence of abdominal obesity, low serum high-density-lipoprotein cholesterol (HDL-C) and diabetes mellitus was lower in subjects who consumed alcohol twice or more per month, with a trend toward reducing the prevalence of metabolic syndrome. Among the current alcohol drinkers, systolic blood pressure, HDL-C, fasting plasma glucose, and prevalence of hypertriglyceridemia tended to increase with increased alcohol consumption. However, low-density-lipoprotein cholesterol concentration, prevalence of abdominal obesity, low serum HDL-C and metabolic syndrome showed the tendency to decrease. Moreover, these statistically significant differences were independent of gender and age. CONCLUSION: Current alcohol consumption is associated with a lower prevalence of metabolic syndrome irrespective of alcohol intake (g/d), and has a favorable influence on HDL-C, waist circumference, and possible diabetes mellitus. However, alcohol intake increases the likelihood of hypertension, hypertriglyceridemia and hyperglycemia. The clinical significance of these findings needs further investigation. PMID- 18416474 TI - Role of the duodenum in regulation of plasma ghrelin levels and body mass index after subtotal gastrectomy. AB - AIM: To investigate the role of the duodenum in the regulation of plasma ghrelin levels and body mass index (BMI), and the correlation between them after subtotal gastrectomy. METHODS: Forty-two patients with T0-1N0-1M0 gastric cancer were divided into two groups after gastrectomy according to digestive reconstruction pattern, Billroth I group (n = 23) and Billroth II group (n = 19). Ghrelin levels were determined with radioimmunoassay (RIA) before and on d 1, 7, 30 and 360 after gastrectomy, and BMI was also measured. RESULTS: The two groups had identical postoperative trends in ghrelin alterations during the early stage, both decreasing sharply to a nadir on d 1 (36.7% vs 35.7%), then markedly increasing on d 7 (51.0% vs 51.1%). On d 30, ghrelin levels in the Billroth I group were slightly higher than those in the Billroth II group. However, those of the Billroth I group recovered to 93.6% on d 360, which approached, although lower than, the preoperative levels, and no statistically significant difference was observed. Those of the Billroth II group recovered to only 81.6% and manifested significant discrepancy with preoperative levels (P = 0.033). Compared with preoperative levels, ghrelin levels of the two groups decreased by 6.9% and 18.4% and BMI fell by 3.3% and 6.4%, respectively. The linear regression correlations were revealed in both groups between decrease of ghrelin level and BMI (R1(2) = 0.297, P = 0.007; R2(2) = 0.559, P < 0.001). CONCLUSION: Anatomically and physiologically, the duodenum compensatively promotes ghrelin recovery and accordingly enhances BMI after gastrectomy. Regarding patients with insufficient ghrelin secretion, ghrelin is positively associated with BMI. PMID- 18416475 TI - Study on vasculogenic mimicry in malignant esophageal stromal tumors. AB - AIM: To investigate whether malignant esophageal stromal tumors contain PAS positive patterned matrix-associated vascular channels, which are lined by tumor cells, but not vascular endothelial cells. That is vasculogenic mimicry (VM) independent of tumor angiogenesis. METHODS: Thirty-six tissue samples of malignant esophageal stromal tumors were analyzed. Tissue sections were stained for Vascular endothelial growth factor (VEGF), CD31 and periodic acid Schiff (PAS). The level of VEGF, the microvascular density (MVD) and the vasculogenic mimicry density (VMD) were determined. RESULTS: PAS-positive patterned matrix associated vascular channels were detected in 33.3% (12/36) of tumor samples. Within these patterned channels, red blood cells were found. The level of VEGF and the MVD in tumors containing patterned channels were significantly higher than those in tumors not containing patterned channels (P < 0.05). At the same time, the malignant degree of tumors was higher, the proportions of tumors containing patterned channels were not only more, but also in the each kind of tumors containing patterned channels. CONCLUSION: In malignant esophageal stromal tumors, a VM mechanism causes some tumor cells to deform themselves and secrete extracellular matrix; thus, PAS-positive patterned matrix-associated vascular channels appear and supplying blood to the tumors to sustain their growth and metastasis. PMID- 18416476 TI - Impact of postoperative omega-3 fatty acid-supplemented parenteral nutrition on clinical outcomes and immunomodulations in colorectal cancer patients. AB - AIM: To investigate the effect of omega-3 fatty acid parenteral supplementation postoperatively on clinical outcomes and immunomodulation in colorectal cancer patients. METHODS: Forty-two patients undergoing radical colorectal cancer resection with an indication for total parenteral nutrition postoperatively were enrolled in this prospective, double-blind, randomized, controlled study. Patients received total parenteral nutrition supplemented with either soybean oil (LCT; Intralipid, Fresenius-Kabi, SO group, n = 21) or a combination of omega-3 fish oil and soybean oil (LCT:fish oil = 5:1, fish oil; Omegaven, Fresenius-Kabi, FO group, n = 21), up to a total of 1.2 g lipid/kg per day for 7 d postoperatively. A same volume calorie and nitrogen was administrated. Routine blood test, biochemistry, systemic levels of IL-6 and TNF-alpha, percentage of CD3+, CD4+, and CD8+ lymphocytes were evaluated preoperatively and on postoperative d 1 and 8. Patient outcome was evaluated considering mortality during the hospital stay, length of postoperative hospital stay, and occurrence of infectious complications. RESULTS: Both lipid regimens were well tolerated. No differences between the two groups were noticed in demographics, baseline blood test, biochemistry, serum levels of IL-6 and TNF-alpha, percentage of CD4+, CD8+ lymphocytes, and ratios of CD4+/CD8+. Compared with those on postoperative d 1, serum IL-6 levels on postoperative d 8 were significantly depressed in the FO group than in the reference group (-44.43 +/- 30.53 vs -8.39 +/- 69.08, P = 0.039). Simultaneously, the ratios of CD4+/CD8+ were significantly increased in the FO group (0.92 +/- 0.62 vs 0.25 +/- 1.22, P = 0.035). In addition, depression of serum TNF-alpha levels (-0.82 +/- 2.71 vs 0.27 +/- 1.67, P = 0.125) and elevation of CD3+ and CD4+ lymphocyte percentage (12.85 +/- 11.61 vs 3.84 +/- 19.62, P = 0.081, 17.80 +/- 10.86 vs 9.66 +/- 17.55, P = 0.084, respectively) were higher in the FO group than in the reference group. Patients in the FO group tended to need a shorter postoperative hospital stay (17.45 +/- 4.80 d vs 19.62 +/- 5.59 d, P = 0.19). No statistically significant difference was found when stratified to mortality and occurrence of infectious complications. CONCLUSION: Postoperative supplementation of omega-3 fatty acids may have a favorable effect on the outcomes in colorectal cancer patients undergoing radical resection by lowering the magnitude of inflammatory responses and modulating the immune response. PMID- 18416477 TI - Analysis of risk factors for the interval time, number and pattern of hepatic metastases from gastric cancer after radical gastrectomy. AB - AIM: To analyze the risk factors for interval time, number and pattern of hepatic metastases from gastric cancer after radical gastrectomy, and provide evidence for predicting and preventing hepatic metastasis from gastric cancer after radical gastrectomy. METHODS: A retrospective study of 87 patients with hepatic metastasis who underwent radical gastrectomy for gastric cancer from 1996 to 2001. The data was analyzed to evaluate significant risk factors for interval time, number and pattern of hepatic metastases originating from gastric cancer after radical gastrectomy. RESULTS: The size of gastric cancer and lymph node metastases were independently correlated with the interval time of hepatic metastases; the depth of invasion was independently correlated with the number of hepatic metastases; while the depth of invasion and Lauren classification were independently correlated with the pattern of hepatic metastases. CONCLUSION: We evaluated the interval time of hepatic metastases with the size of gastric cancer and lymph node metastases. The depth of invasion could be used to evaluate the number of hepatic metastases, while the depth of invasion and the Lauren classification could be used to evaluate the pattern of hepatic metastases in patients who underwent radical gastrectomy. PMID- 18416478 TI - Ulcerative colitis presenting as leukocytoclastic vasculitis of skin. AB - A number of cutaneous changes are known to occur in the course of inflammatory bowel diseases (IBD), including pyoderma gangrenosum, erythema nodosum, perianal disease, erythematous eruptions, urticaria, and purpura. However, occurrence of skin manifestations prior to the development of ulcerative colitis is a rare occasion. Here, we report a case of ulcerative colitis associated with leukocytoclastic vasculitis in which the intestinal symptoms became overt 8 mo after the development of skin lesions. PMID- 18416479 TI - Association of primary biliary cirrhosis with idiopathic thrombocytopenic purpura. AB - Although both primary biliary cirrhosis (PBC) and idiopathic thrombocytopenic purpura (ITP) are autoimmune diseases, the association of the 2 diseases is rare. Here, we report a case of ITP that developed during the follow-up of PBC in a 74 year-old man. The patient had been diagnosed with PBC 12 years previously, and had received treatment with ursodeoxycholic acid. The platelet count decreased from approximately 60 x 10(9)/L to 8 x 10(9)/L, and the association of decompensated liver cirrhosis (PBC) with ITP was diagnosed. Steroid and immune gamma globulin therapy were successful in increasing the platelet count. Interestingly, human leukocyte antigen genotyping detected the alleles DQB1*0601 and DRB1*0803, which are related to both PBC and ITP in Japanese patients. This case suggests common immunogenetic factors might be involved in the development of PBC and ITP. PMID- 18416480 TI - Non-invasive prediction of oesophageal varices in cirrhosis. AB - Non-invasive predictors of varices in cirrhosis would reduce the need for screening endoscopies. Platelet count and spleen size have been shown to be useful parameters, in mixed groups of cirrhotics with different aetiologies. We evaluated this in two homogeneous groups with cirrhosis due to hepatitis C and alcohol. Non-invasive predictors appear promising in the former group, but less so in the latter group. PMID- 18416481 TI - Breaking the mirror: Asymmetrical disconnection between the phonological input and output codes. AB - In this paper, we study the link between the processing systems that sustain speech perception and production in a patient (F.A.) with conduction aphasia. Her pattern of performance in repetition task - quantitative but also qualitative striking difference in errors with pseudowords versus words - cannot be properly accounted for either by a perception deficit or by a production deficit. We discuss this finding according to theoretical models of phonological processing and show that it is best explained by an impaired ability to transfer phonological information from the perception to the production system. We also probed for a phonological link in the opposite direction, from the production to the perception system. F.A.'s results show that this link was not impaired. Overall, our results suggest that (a) the phonological codes in perception and in production are separate but connected by two conversion mechanisms and that (b) these two mechanisms can be disrupted independently. PMID- 18416482 TI - Models of confabulation: a critical review and a new framework. AB - Confabulation can be defined as statements or actions that involve distortions of memories. This paper reviews current theories of confabulation focusing on source monitoring, temporal-context, and retrieval theories. The attributes and criticisms of these three models are discussed. From this review, a three-factor cognitive-neuropsychological framework is proposed, which can be used to explain the variable symptoms of confabulation. The framework takes its basis from the Langdon and Coltheart (2000a, 2000b) cognitive model of delusional belief formation. The model suggests that two deficits are likely in most cases of confabulation - an executive control retrieval deficit and an evaluation deficit. It also takes into consideration how the general organization of the autobiographical memory store and a person's individual emotional/motivational biases can influence confabulatory symptoms and content. This is an overarching framework that can be used to model confabulations, and it builds upon links between delusions and confabulation. PMID- 18416483 TI - Lexical acquisition in progressive aphasia and frontotemporal dementia. AB - We investigated the characteristics of language difficulty in frontotemporal dementia (FTD) by exposing these patients to a new verb in a naturalistic manner and then assessing acquisition of the grammatical, semantic, and thematic matrix information associated with the new word. We found that FTD patients have difficulty relative to healthy seniors in their acquisition of the new verb, but that progressive nonfluent aphasia (PNFA), semantic dementia (SD), and social/dysexecutive variant (SOC/EXEC) subgroups of FTD demonstrate relatively distinct impairment profiles. Specifically, PNFA patients showed relative difficulty assigning the new verb to its correct grammatical form class, reflecting compromised processing of the associated grammatical information. SD patients were impaired at associating the new word with its pictorial representation, suggesting impaired processing of the new verb's semantic attributes. SOC/EXEC patients showed their greatest difficulty judging violations of the new word's associated thematic roles, implying that limited executive resources underlie in part the difficulty in integrating grammatical and semantic information into a coherent thematic matrix. Similar impairment profiles were seen during a follow-up session one week after the initial evaluation. These deficits in lexical acquisition reflect the breakdown of a language-processing system that consists of highly interactive but partially dissociable grammatical, semantic, and resource-based components, leading to relatively distinct language processing deficits in each subgroup of patients with FTD. PMID- 18416484 TI - Local capture in Balint's syndrome: effects of grouping and item familiarity. AB - We assessed the factors contributing to local capture in G.K., a patient with Balint's syndrome. G.K. found it difficult to identify the global shape of a compound stimulus, and his poor identification of the global shape was not resolved by introducing familiar local stimuli that were either from the same category as the global target but not tied to a response (Experiment 1) or that belonged to a different category to the target (Experiment 2). Effects of item familiarity were tested by examining local capture from Hebrew letters on compound English letters. Prior to training with the Hebrew letters, G.K. was able to identify the global compounds (Experiment 3a). In contrast, after training with the letters, there was a significant increase in local capture (Experiment 3b). Two additional experiments examined effects of local grouping. Local capture was reduced when the local elements were rectangles or ellipses and grouped by closure of local elements (Experiment 4). Global discrimination also improved when local items were unfamiliar rectangular figure eights (Experiment 5a). However, these grouping effects were counteracted by effects of familiarity when the local elements were changed slightly to become familiar number eights (Experiment 5b). The data suggest that familiar local items are likely to capture spatial attention, whilst grouping by closure between local elements can help global shapes compete for spatial attention with local elements in such patients. The impairment in global perception in Balint's syndrome is not all-or-none but graded according to the factors influencing local capture. PMID- 18416485 TI - What does a patient with semantic dementia remember in verbal short-term memory? Order and sound but not words. AB - In this study, we explored capacities for three different aspects of short-term verbal memory in patients with semantic dementia. As expected, the two patients had poor recall for lexico-semantic item information, as assessed by immediate serial recall of word lists. In contrast, their short-term memory for phonological information was preserved, as evidenced by normal performance for immediate serial recall of nonword lists, with normal or increased nonword phonotactic-frequency effects, and increased sensitivity to phonological lures in a delayed probe recognition task. Furthermore, the patients appeared to have excellent memory for the serial order of the words in a list. These data provide further support for the proposal that language knowledge is a major determining factor of verbal STM capacity, but they also highlight the necessary distinction of processes involved in item and order recall, as proposed by recent models of STM. PMID- 18416486 TI - Colour agnosia impairs the recognition of natural but not of non-natural scenes. AB - Scene recognition can be enhanced by appropriate colour information, yet the level of visual processing at which colour exerts its effects is still unclear. It has been suggested that colour supports low-level sensory processing, while others have claimed that colour information aids semantic categorization and recognition of objects and scenes. We investigated the effect of colour on scene recognition in a case of colour agnosia, M.A.H. In a scene identification task, participants had to name images of natural or non-natural scenes in six different formats. Irrespective of scene format, M.A.H. was much slower on the natural than on the non-natural scenes. As expected, neither M.A.H. nor control participants showed any difference in performance for the non-natural scenes. However, for the natural scenes, appropriate colour facilitated scene recognition in control participants (i.e., shorter reaction times), whereas M.A.H.'s performance did not differ across formats. Our data thus support the hypothesis that the effect of colour occurs at the level of learned associations. PMID- 18416487 TI - The impact of progressive semantic loss on reading aloud. AB - The role of semantics in reading aloud remains controversial. To explore this issue, the current study examined the impact of semantic loss on reading-aloud performance in 7 patients with semantic dementia. The results revealed a heterogenous pattern of reading difficulties. Of the patients, 2 selectively made errors on inconsistent words (i.e., surface dyslexia), 4 had a generalized reading deficit with increased errors on consistent words, inconsistent words, and nonwords, while the remaining patient had relatively intact reading-aloud accuracy. All patients had longer reading latencies on real words than controls. The relationship between the reading and semantic deficits in the patients was examined at the item-specific level. This suggested that reading-aloud errors were related to the semantic impairment for inconsistent words but not consistent words. In contrast, semantic loss was related to longer latencies for both consistent and inconsistent words. These findings support models of reading that include a role for semantics in the reading-aloud process. PMID- 18416488 TI - Maintaining the ties that bind: the role of an intermediate visual memory store in the persistence of awareness. AB - Segregation and feature binding are essential to the perception and awareness of objects in a visual scene. When a fragmented line-drawing of an object moves relative to a background of randomly oriented lines, the previously hidden object is segregated from the background and consequently enters awareness. Interestingly, in such shape-from-motion displays, the percept of the object persists briefly when the motion stops, suggesting that the segregated and bound representation of the object is maintained in awareness. Here, we tested whether this persistence effect is mediated by capacity-limited working-memory processes, or by the amount of object-related information available. The experiments demonstrate that persistence is affected mainly by the proportion of object information available and is independent of working-memory limits. We suggest that this persistence effect can be seen as evidence for an intermediate, form based memory store mediating between sensory and working memory. PMID- 18416489 TI - Localizing the deficit in a case of jargonaphasia. AB - We report the case of a neologistic jargonaphasic and ask whether her target related and abstruse neologisms are the result of a single deficit, which affects some items more severely than others, or two deficits: one to lexical access and the other to phonological encoding. We analyse both correct/incorrect performance and errors and apply both traditional and formal methods (maximum-likelihood estimation and model selection). All evidence points to a single deficit at the level of phonological encoding. Further characteristics are used to constrain the locus still further. V.S. does not show the type of length effect expected of a memory component, nor the pattern of errors associated with an articulatory deficit. We conclude that her neologistic errors can result from a single deficit at a level of phonological encoding that immediately follows lexical access where segments are represented in terms of their features. We do not conclude, however, that this is the only possible locus that will produce phonological errors in aphasia, or, indeed, jargonaphasia. PMID- 18416490 TI - Examination of the split fovea theory in a case of pure left hemialexia. AB - To address the extent to which the visual foveal representation is split, we examine the case of a patient, M.B., suffering from a left mesial occipital lesion and presenting a pure left hemialexia and a right hemianopia with a spared area of the macula. Reading performance on tachistoscopically presented four letter words and pseudowords in the spared area of the right visual field was significantly better than reading performance in the intact left visual field. Reading performance in the spared area of the right visual field was also significantly better than reading performance of stimuli centred on the fovea. Moreover, a length effect was found only in the left half of pseudowords centred on the fovea, but not in the right half (up to five letters). These differences in reading efficiency between the left and right halves of the foveal region militates in favour of the split fovea theory and cannot be explained by the bilateral projection theory. PMID- 18416491 TI - Visual attention in adults with developmental dyslexia: evidence from manual reaction time and saccade latency. AB - Two studies were conducted to investigate visual attention deficits in dyslexia. In Experiment 1, adults with dyslexia and age- and IQ-matched controls completed a simple cueing task; participants responded to briefly presented (20 ms) eccentric targets (3 degrees , 6 degrees , or 9 degrees ) with a key press. In Experiment 2, the same participants completed a saccade version of the task, and saccade amplitude, accuracy, and latency were measured. The results revealed comparable performance between the groups on the manual reaction time task. The groups also performed similarly in saccade accuracy and latency. Moreover, neither group showed a visual field asymmetry in their performance, with the exception that adults with dyslexia showed longer saccade latency for 9 degrees targets presented to their left visual field than did controls. However, on the latter measure, the majority (78%) of those with dyslexia performed within the range of the control group. Correlational analyses revealed associations between reading and phoneme awareness in both groups, but phoneme awareness was not associated with visual attention in adult dyslexics. Together, the results are not compatible with a visual attention deficit in adult dyslexia, while they provide support for the phonological deficit hypothesis. PMID- 18416492 TI - Maximizing the power of comparing single cases against a control sample: an argument, a program for making comparisons, and a worked example from the Pyramids and Palm Trees Test. AB - In neuropsychological research, it is frequently necessary to compare the performance of a single case with that of a control sample. Recently, there has been a discussion about whether existing methods are adequate in preventing Type 1 errors due to increased variability in the single-case data (Crawford, Garthwaite, Howell, & Gray, 2004; Mitchell, Mycroft, & Kay, 2004; Mycroft, Mitchell, & Kay, 2002) and about the power of any comparisons. In this paper, we propose an extension of the modified t test introduced by Crawford and Howell (1998). We provide Monte Carlo simulations that show that it is possible to increase the power of the modified t test by retesting the single case. We also make available a computer program that implements the method introduced in this paper and that can be used by neuropsychologists to test for a difference between a single case and a control group. We apply our procedure to an example of a patient examined on the "Pyramids and Palm Trees" test. PMID- 18416493 TI - Processing of syntactically complex sentences relies on verbal short-term memory: evidence from a short-term memory patient. AB - The role of the phonological loop in auditory sentence comprehension was examined in a 35-year-old woman with a selective deficit of verbal short-term memory (STM). More specifically, the objective of the experiment was to test whether sentence comprehension is limited by number of propositions, as suggested by Rochon, Waters, and Caplan (2000), or whether it depends on syntactic complexity. In an offline task, severe impairment was present on cleft objects and centre embedded structures; the deficit on object relatives in right peripheral position was less relevant, and the patient was able to handle sentential coordination easily. In an online task, her processing of centre-embedded structures and object relatives in right peripheral position was significantly slower than that in controls. She was also significantly slower in processing the verb of the object relative in centre-embedded structures. The results obtained do not support the claim that the difficulties encountered by patients with limited working-memory resources are due to the number of propositions in the sentence; they do, however, suggest a direct involvement of the phonological loop in processing syntactically complex sentences. PMID- 18416494 TI - Semantic errors in deep dyslexia: does orthographic depth matter? AB - Semantic errors of oral reading by aphasic patients are said to be comparatively rare in languages with a shallow orthography. The present report concerns three bilingual brain-damaged patients who prior to their stroke were fluent in both English, an orthographically deep language, and Welsh, an orthographically shallow language. On a picture-naming task, each patient made a similar proportion of semantic errors in the two languages. Similarly, in oral reading of the corresponding words, no patient produced proportionally more semantic paralexias in English than in Welsh. The findings are discussed in relation to the summation hypothesis as invoked by Miceli, Capasso, and Caramazza (1994) to explain apparent differences in frequency of semantic errors of reading in languages differing in orthographic depth. PMID- 18416495 TI - The influence of spatial coordinates in a case of an optic ataxia-like syndrome following cerebellar and thalamic lesion. AB - We report the case of a patient (S.C.) who, after a lesion involving the midbrain, thalamus, and cerebellum on the left side, was unable to reach a target at which he was not directly gazing. When attempting to do so with either hand, he showed an optic ataxia-like behaviour: A rightward deviation was manifest with respect to the real position of the target object, accuracy being worse for the right than for the left hand and for the right than for the left hemifield. To assess whether S.C.'s reaching accuracy was affected by the position of the target with respect to the retina, eyes, and/or head, he was asked to perform a pointing task in different conditions. By manipulating eyes and head position, the relative location of the target with respect to these body parts was altered. Further, to verify the relevance of the visual feedback for the accuracy of the reaching responses, the task was also performed either with or without vision of the performing hand (closed- and open-loop conditions). The results showed that the patient's reaching impairment reflects a deficit in spatio-motor coding mainly within head-centred coordinates. The lack of visual feedback about the hand spatial location worsened the reaching performance. This new pattern of reaching deficits is discussed in relation to the theoretical framework of both parietal optic ataxia and cerebellar ataxia. PMID- 18416496 TI - Comparison of a single case to a control or normative sample in neuropsychology: development of a Bayesian approach. AB - Frequentist methods are available for comparison of a patient's test score (or score difference) to a control or normative sample; these methods also provide a point estimate of the percentage of the population that would obtain a more extreme score (or score difference) and, for some problems, an accompanying interval estimate (i.e., confidence limits) on this percentage. In the present paper we develop a Bayesian approach to these problems. Despite the very different approaches, the Bayesian and frequentist methods yield equivalent point and interval estimates when (a) a case's score is compared to that of a control sample, and (b) when the raw (i.e., unstandardized) difference between a case's scores on two tasks are compared to the differences in controls. In contrast, the two approaches differ with regard to point estimates of the abnormality of the difference between a case's standardized scores. The Bayesian method for standardized differences has the advantages that (a) it can directly evaluate the probability that a control will obtain a more extreme difference score, (b) it appropriately incorporates error in estimating the standard deviations of the tasks from which the patient's difference score is derived, and (c) it provides a credible interval for the abnormality of the difference between an individual's standardized scores; this latter problem has failed to succumb to frequentist methods. Computer programs that implement the Bayesian methods are described and made available. PMID- 18416497 TI - Two orienting mechanisms in posterior parietal lobule: an rTMS study of the Simon and SNARC effects. AB - The aim of the present study was to investigate, with repetitive transcranial magnetic stimulation (rTMS), the involvement of posterior parietal lobule (PPL) circuits in producing the Simon effect and the spatial-numerical association of response codes (SNARC) effect. The Simon effect is indexed by faster responses to left-side stimuli with left- than with right-side key-presses and faster responses to right-side stimuli with right- than with left-side key-presses. The SNARC effect is indexed by faster responses to smaller numbers with left- than with right-side key-presses and faster responses to larger numbers with right- than with left-side key-presses. Participants were required to perform a parity judgement on numbers ranging from 1 to 9 (without 5), by pressing a left or a right response key. The numbers were presented to either the left or the right side of fixation. Two bilateral PPL areas lying along the intraparietal sulcus (IPS) were stimulated with rTMS. Results suggested a causal role of the anterior portion of PPL of both hemispheres in the Simon effect and of the posterior portion of PPL of both hemispheres in the Simon effect and the SNARC effect. PMID- 18416498 TI - Qualitatively different forms of pure alexia. AB - In this study we investigated two patients with pure alexia, F.C. and L.D.S., in order to make inferences about how processes and levels involved in the early stage of visual word recognition are organized and how they can be selectively damaged. Moreover, we investigated whether pure alexia can be caused by different functional deficits. F.C. and L.D.S. were presented with tasks of letter processing and tasks of orthographic integration. There was a clear double dissociation between the pattern of performance of F.C. and L.D.S. F.C. was able to process single letters rapidly and accurately, but was unable to group together the letters that he had correctly identified. By contrast, L.D.S. was slower and more impaired at letter identification, but she could use letter groups to assist reading. Thus, two different forms of pure alexia emerged: F.C. has a higher level deficit in integrating letters, whereas L.D.S. has a lower level deficit in letter processing. The results support the assumption of a functional organization of the reading process that involves a series of orthographic units (i.e., single letters, sublexical letter groups, and the lexical unit), which can be selectively damaged. Finally, our data present difficulties for models of pure alexia that assume all patients to have a low level processing deficit. PMID- 18416499 TI - Family resemblance: ten family members with prosopagnosia and within-class object agnosia. AB - We report on neuropsychological testing done with a family in which many members reported severe face recognition impairments. These 10 individuals were high functioning in everyday life and performed normally on tests of low-level vision and high-level cognition. In contrast, they showed clear deficits with tests requiring face memory and judgements of facial similarity. They did not show deficits with all aspects of higher level visual processing as all tested performed normally on a challenging facial emotion recognition task and on a global-local letter identification task. On object memory tasks requiring recognition of particular cars and guns, they showed significant deficits so their recognition impairments were not restricted to facial identity. These results strongly suggest the existence of a genetic condition leading to a selective deficit of visual recognition. PMID- 18416500 TI - The orthographic buffer in writing Chinese characters: evidence from a dysgraphic patient. AB - We investigated the postlexical processes in writing Chinese characters by studying the delayed copying performance of a Chinese dysgraphic patient, W.L.Z. His delayed copying difficulty could not be attributed to peripheral motor deficit and could not be readily explained by lexical or semantic factors. Instead, the copying performance was sensitive to a word length variable (number of logographemes), and the most prevalent errors were logographeme substitutions. Furthermore, in the substitution errors, the target logographemes and responses tended to share visual/motoric attributes. We propose that the delayed copying difficulty reflects a deficit to the buffering component in writing (coined "logographeme output buffer"), and the universality and language-specific features of the output buffer in writing are discussed. PMID- 18416501 TI - I can't recognize your face but I can recognize its movement. AB - Idiosyncratic facial movements can provide a route to facial identity (review in Roark, Barrett, Spence, Abdi, & O'Toole, 2003). However, it is unclear whether recognizing a face in this way involves the same cognitive or neural mechanisms that are involved in recognizing a static face. Three studies on a developmental prosopagnosic (C.S.) showed that although he is impaired at recognizing static faces, he can discriminate between dynamic identities (Experiments 1a and 1b) and can learn to name individuals on the basis of their idiosyncratic facial movements (Experiment 2), at levels that are comparable to those of matched and undergraduate control groups. These results suggest a possible cognitive dissociation between mechanisms involved in dynamic compared to static face recognition. However, future work is needed to fully understand this dissociation. PMID- 18416502 TI - Dissociation between deficits in explicit procedures and implicit processes in the visual-spatial and the phonological systems during reading acquisition. AB - Two clinical studies are reported of children with proficient word-reading skill despite severe performance deficits of explicit visual recognition of the lateral ("mirror image") orientation of letters. In dissociation from their deficit in such an explicit procedure, the children had proficient implicit processing of letters that change identity with lateral reversal, as shown in nonword reading, and in letter-naming accuracy and reaction times relative to a normal comparison sample of children. In one child, another dissociation was also apparent in the phonological system. He had a severe performance deficit in explicit processing of phonemes (phonemic awareness) but was proficient in grapheme-phoneme recoding for reading. The findings indicate that it is possible for both the visual spatial coding and the phonological systems to function in reading acquisition with impaired performance of explicit procedures in each system while maintaining proficient development of reading skill. PMID- 18416504 TI - Evidence of a divided-attention advantage in autism. AB - People with autism spectrum disorders appear to have some specific advantages in visual processing, including an advantage in visual search tasks. However, executive function theory predicts deficits in tasks that require divided attention, and there is evidence that people with autism have difficulty broadening their attention (Mann & Walker, 2003). We wanted to know how robust the known attentional advantage is. Would people with autism have difficulty dividing attention between central and peripheral tasks, as is required in the Useful Field of View task, or would they show an advantage due to strengths in visual search? Observers identified central letters and localized peripheral targets under both focused- and divided-attention conditions. Participants were 20 adults with high-functioning autism and Asperger's syndrome and 20 adults matched to the experimental group on education, age, and IQ. Contrary to some predictions, individuals with autism tended to show relatively smaller divided attention costs than did matched adults. These results stand in stark contrast to the predictions of some prevalent theories of visual and cognitive processing in autism. PMID- 18416503 TI - Nouns, verbs, objects, actions, and the animate/inanimate effect. AB - We report an aphasic patient, Z.B.L., who showed a significant advantage for verbs compared to nouns in picture-naming tests. Within the object class, he performed better on animate things than on nonliving things in picture naming as well as in an "attribute judgement task". This pattern of performance is contrary to the central prediction of a recent proposal (Bird, Howard, & Franklin, 2000), which attributes noun-verb dissociation in aphasic patients to deficits in processing certain kinds of semantic features. This model proposes that conceptual representations of verbs have a lower proportion of sensory features than do representations of nouns; the same is proposed for inanimate versus animate items within the noun category. Noun deficits are assumed to arise due to impairment for the processing of sensory features. The model predicts that if a patient is more impaired for nouns than for verbs, he will also display more difficulty with animate than with inanimate objects. Contrary to predications derived from this theory, Z.B.L. performed better with animate than inanimate nouns. PMID- 18416505 TI - Morphological processing with deficient phonological short-term memory. AB - This paper investigates the processing of Hebrew derivational morphology in an individual (S.E.) with deficient phonological short-term memory. In comparison to 10 age- and education-matched men, S.E. was impaired on digit span tasks and demonstrated no recency effect in word list recall. S.E. had low word retention span, but he exhibited phonological similarity and word length effects. His ability to make lexical decisions was intact. In a paired-associate test S.E. successfully learned semantically and morphologically related pairs but not phonologically related pairs, and his learning of nonwords was facilitated by the presence of Hebrew consonant roots. Semantic and morphological similarity enhanced immediate word recall. Results show that S.E. is capable of conducting morphological decomposition of Hebrew-derived words despite his phonological deficit, suggesting that transient maintenance of morphological constituents is independent of temporary storage and rehearsal of phonological codes, and that each is processed separately within short-term memory. PMID- 18416506 TI - Oculomotor capture in ADHD. AB - It is generally thought that deficits in response inhibition form an important area of dysfunction in patients with attention-deficit/hyperactivity disorder (ADHD). However, recent research using visual search paradigms seems to suggest that these inhibitory deficits do not extend towards inhibiting irrelevant distractors. Using an oculomotor capture task, the present study investigated whether boys with ADHD and their nonaffected brothers are impaired in suppressing reflexive eye movements to a task-irrelevant onset distractor. Results showed that boys with ADHD had slower responses than controls, but were as accurate in their eye movements as controls. Nonaffected brothers showed similar problems in the speed of responding as their affected brothers, which might suggest that this deficit relates to a familial risk for developing the disorder. Importantly, all three groups were equally captured by the distractor, which shows that boys with ADHD and their brothers are not more distracted by the distractor than are controls. Saccade latency and the proportion of intrusive saccades were related to continuous dimensions of ADHD symptoms, which suggests that these deficits are not simply present or absent, but rather indicate that the severity of these deficits relate to the severity of ADHD. The finding that boys with ADHD (and their nonaffected brothers) did not have problems inhibiting irrelevant distractors contradicts a general response inhibition deficiency in ADHD, which may be explained by the relatively independency of working memory in this type of response inhibition. PMID- 18416507 TI - Local bias and local-to-global interference without global deficit: a robust finding in autism under various conditions of attention, exposure time, and visual angle. AB - A wide variety of paradigms have shown that autistic individuals present with superior performance on visual tasks. Here, the impact of task constraints on visual hierarchical processing in autism was investigated. By employing free- and forced-choice procedures, global and local processing of Navon-type hierarchical numerals was examined in 15 autistic persons (13 males, 2 females) and a comparison group. In the free-choice condition, autistics chose global and local targets randomly, though they were faster responding to local than to global targets, regardless of visual angle and exposure duration. In contrast, the comparison group exhibited a global advantage in naming time, which was evident only for shorter exposures, as well as effects of visual angle. In the forced choice condition, autistics presented with a more important local-to-global interference than global-to-local interference, whereas the comparison group exhibited global advantage and bidirectional interference. Overall, the autistic participants presented with atypical local-to-global interference and local advantage in incongruent conditions (where global and local targets differ), in naming time as well as accuracy. The relative insensitivity of local bias to task constraints in autistics, in comparison to nonautistic participants, indicates that local bias, with local-to-global interference, is a key and characteristic feature of autistic visual cognition and a strong candidate for the "endophenotype" of autism. PMID- 18416508 TI - Do local bias and local-to-global interference reflect intact global processing in autism? PMID- 18416510 TI - Semantic priming in deep-phonological dyslexia: contrasting effects of association and similarity upon abstract and concrete word reading. AB - We report a series of experiments investigating the reading abilities of a patient R.O.M. who exhibited the syndrome of deep-phonological dyslexia. In a series of semantic priming tasks, R.O.M. was requested to read word lists containing either abstract or concrete words, which were related by semantic similarity or semantic association, or which shared no clear semantic relationship. R.O.M. read semantically similar concrete words significantly more accurately than unrelated items, but showed no such advantage for semantically associated concrete words. By contrast, semantically associated abstract words were read significantly more accurately than unrelated items, but there was no evidence of priming for semantically similar abstract words. Thus, we describe an attempt to harness semantic priming to find converging evidence for a distinction between the representational frameworks underpinning abstract and concrete concepts. The findings are considered in the context of competing theories of the abstract/concrete distinction. PMID- 18416511 TI - Harmonic priming in an amusic patient: the power of implicit tasks. AB - Our study investigated with an implicit method (i.e., priming paradigm) whether I.R. - a brain-damaged patient exhibiting severe amusia - processes implicitly musical structures. The task consisted in identifying one of two phonemes (Experiment 1) or timbres (Experiment 2) on the last chord of eight-chord sequences (i.e., target). The targets were harmonically related or less related to the prior chords. I.R. displayed harmonic priming effects: Phoneme and timbre identification was faster for related than for less related targets (Experiments 1 and 2). However, I.R.'s explicit judgements of completion for the same sequences did not differ between related and less related contexts (Experiment 3). Her impaired performance in explicit judgements was not due to general difficulties with task demands since she performed like controls for completion judgements on spoken sentences (Experiment 4). The findings indicate that implicit knowledge of musical structures might remain intact and accessible, even when explicit judgements and overt recognition have been lost. PMID- 18416512 TI - Preorthographical constraints on visual word recognition: evidence from a case study of developmental surface dyslexia. AB - We investigated the visual word recognition ability of M.T., a young boy with surface dyslexia, by means of a paradigm that measures performance as a function of the eye fixation position within the word, known as the "viewing-position effect" paradigm. In well-achieving readers, the viewing-position effect is mainly determined by factors affecting letter visibility and by lexical constraints on word recognition. We further quantified M.T.'s sensory limitations on letter visibility by computing visual-span profiles - that is, the number of letters recognizable at a glance. Finally, in an ideal-observer's perspective, M.T.'s performance was compared with a parameter-free model combining M.T.'s letter visibility data with a simple lexical matching rule. The results showed that M.T. did not use the whole visual information available on letter identities to recognize words and that preorthographical factors constrained his word recognition performance. The results can be best accounted for by a reduction of the number of letters processed in parallel. PMID- 18416513 TI - The representation of unseen objects in visual neglect: effects of view and object identity. AB - We provide evidence for long-term priming based on view-specific representations of neglected stimuli. A patient with visual neglect, M.P., was asked to search for a target presented amongst other objects on a table. Subsequently recognition memory was tested for items that were identified and for items missed in search. Items that were missed were rejected more slowly than novel items in the recognition memory task, providing evidence for implicit processing (Experiment 1). Implicit memory for missed items was both item-specific (Experiment 2) and view-specific (Experiment 3), and it was eliminated when there were intervening activities lasting about 1 hour (Experiment 4). There was also an implicit memory for distractors in the search task, which was item- but not view-specific (Experiments 2 and 3) and it lasted for at least an hour, even with other intervening activities (Experiment 4). The data suggest that the representations of neglected stimuli may differ qualitatively from those of nonneglected items, with representations of neglected objects being both view-specific and vulnerable across extended retention intervals. The results support the argument that attention is needed in order to encode object representations that are robust to view transformations and temporal decay or interference. PMID- 18416514 TI - Computers and accessibility. PMID- 18416515 TI - Individual responses to a method of cursor assistance. AB - PURPOSE: The aim of this study was to evaluate a new click assistance technique, Steady Clicks, designed to help computer users with motor impairments to click more accurately using a mouse. Specifically, Steady Clicks suppresses two types of errors: slipping while clicking and accidentally clicking. Steady Clicks suppresses these errors by freezing the cursor during mouse clicks, preventing overlapping button presses and suppressing clicks made while the mouse is moving at a high velocity. METHOD: Eleven individuals with motor impairments participated in a repeated-measures evaluation of Steady Clicks. This evaluation involved performing a clicking task both with and without Steady Clicks, while time-stamped log files detailing each participant's cursor movements and mouse button presses were recorded. RESULTS: When using Steady Clicks, five of the 11 participants were able to select targets using significantly fewer attempts, and had significantly improved overall task performance times. Blocking of overlapping and high velocity clicks also shows promise as an error filter. For some participants, Steady Clicks had effects beyond a simple reduction in errors and error correction time, including faster positioning times with fewer, shorter pauses. Two participants slipped a greater distance when using Steady Clicks, suggesting they were taking advantage of the support. Two were much less fatigued, and one was able to start using a simpler clicking strategy. Nine participants preferred Steady Clicks to the unassisted condition. CONCLUSION: Large individual differences were found not only in performance but also in the ways that individuals reacted to and benefited from the cursor assistance. The results showed that Steady Clicks can be used to effectively block slipping and accidental clicks by individuals for whom this is a problem. This assistance could be used in conjunction with existing techniques for cursor positioning, to enable faster and more effective mouse use for those who currently struggle with the standard computer mouse. PMID- 18416516 TI - The Vocal Joystick: evaluation of voice-based cursor control techniques for assistive technology. AB - PURPOSE: Mouse control has become a crucial aspect of many modern day computer interactions. This poses a challenge for individuals with motor impairments or those whose use of hands is restricted due to situational constraints. We present a system called the Vocal Joystick which allows the user to continuously control the mouse cursor by varying vocal parameters such as vowel quality, loudness and pitch. METHOD: Evaluations were conducted to characterize expert performance capability of the Vocal Joystick, and to compare novice user performance and preference for the Vocal Joystick and two other existing speech based cursor control methods. RESULTS: Our results show that Fitts' law, a well adopted model of human motor performance for movement tasks, is a good predictor of the speed - accuracy tradeoff for the Vocal Joystick, and suggests that the optimal performance of the Vocal Joystick may be comparable to that of a conventional hand-operated joystick. Novice user evaluations show that the Vocal Joystick can be used by people without extensive training, and that it presents a viable alternative to existing speech-based cursor control methods. CONCLUSIONS: The Vocal Joystick, with its ease of use, minimal setup requirement, and controllability, offers promise for providing an efficient method for cursor control and other forms of continuous input for individuals with motor impairments. PMID- 18416517 TI - Enabling devices, empowering people: the design and evaluation of Trackball EdgeWrite. AB - PURPOSE: To describe the research and development that led to Trackball EdgeWrite, a gestural text entry method that improves desktop input for some people with motor impairments. To compare the character-level version of this technique with a new word-level version. Further, to compare the technique with competitor techniques that use on-screen keyboards. METHOD: A rapid and iterative design-and-test approach was used to generate working prototypes and elicit quantitative and qualitative feedback from a veteran trackball user. In addition, theoretical modelling based on the Steering law was used to compare competing designs. RESULTS: One result is a refined software artifact, Trackball EdgeWrite, which represents the outcome of this investigation. A theoretical result shows the speed benefit of word-level stroking compared to character-level stroking, which resulted in a 45.0% improvement. Empirical results of a trackball user with a spinal cord injury indicate a peak performance of 8.25 wpm with the character level version of Trackball EdgeWrite and 12.09 wpm with the word-level version, a 46.5% improvement. Log file analysis of extended real-world text entry shows stroke savings of 43.9% with the word-level version. Both versions of Trackball EdgeWrite were better than on-screen keyboards, particularly regarding user preferences. Follow-up correspondence shows that the veteran trackball user with a spinal cord injury still uses Trackball EdgeWrite on a daily basis 2 years after his initial exposure to the software. CONCLUSIONS: Trackball EdgeWrite is a successful new method for desktop text entry and may have further implications for able-bodied users of mobile technologies. Theoretical modelling is useful in combination with empirical testing to explore design alternatives. Single-user lab and field studies can be useful for driving a rapid iterative cycle of innovation and development. PMID- 18416518 TI - Real and virtual explorations of the environment and interactive tracking of movable objects for the blind on the basis of tactile-acoustical maps and 3D environment models. AB - PURPOSE.: This study describes the development of a multi-functional assistant system for the blind which combines localisation, real and virtual navigation within modelled environments and the identification and tracking of fixed and movable objects. METHOD: The approximate position of buildings is determined with a global positioning sensor (GPS), then the user establishes exact position at a specific landmark, like a door. This location initialises indoor navigation, based on an inertial sensor, a step recognition algorithm and map. Tracking of movable objects is provided by another inertial sensor and a head-mounted stereo camera, combined with 3D environmental models. This study developed an algorithm based on shape and colour to identify objects and used a common face detection algorithm to inform the user of the presence and position of others. RESULTS: The system allows blind people to determine their position with approximately 1 metre accuracy. Virtual exploration of the environment can be accomplished by moving one's finger on a touch screen of a small portable tablet PC. The name of rooms, building features and hazards, modelled objects and their positions are presented acoustically or in Braille. CONCLUSIONS: Given adequate environmental models, this system offers blind people the opportunity to navigate independently and safely, even within unknown environments. Additionally, the system facilitates education and rehabilitation by providing, in several languages, object names, features and relative positions. PMID- 18416519 TI - Indoor wayfinding: developing a functional interface for individuals with cognitive impairments. AB - PURPOSE: Assistive technology for wayfinding will significantly improve the quality of life for many individuals with cognitive impairments. The user interface of such a system is as crucial as the underlying implementation and localisation technology. We studied the user interface of an indoor navigation system for individuals with cognitive impairments. METHOD: We built a system using the Wizard-of-Oz technique that let us experiment with many guidance strategies and interface modalities. Through user studies, we evaluated various configurations of the user interface for accuracy of route completion, time to completion, and user preferences. We used a counter-balanced design that included different modalities (images, audio, and text) and different routes. RESULTS: We found that although users were able to use all types of modalities to find their way indoors, they varied significantly in their preferred modalities. We also found that timing of directions requires careful attention, as does providing users with confirmation messages at appropriate times. CONCLUSIONS: Our findings suggest that the ability to adapt indoor wayfinding devices for specific users' preferences and needs will be particularly important. PMID- 18416520 TI - A voice-activated integrated development environment for manually disabled programmers. AB - PURPOSE: The paper presents the design and implementation of voice-based interface to support two types of application development activities (coding, and visual GUI design) by manually disabled programmers. METHOD: Prototype programs were implemented, and tests were performed for both proposed voice-based interface models. RESULTS: The interface design concepts, and the results and lessons learned of the usability studies are discussed. CONCLUSIONS: The usability studies indicated that the interface models were sound, although the voice recognition aspects left something to be desired. PMID- 18416521 TI - MobileASL: intelligibility of sign language video over mobile phones. AB - For Deaf people, access to the mobile telephone network in the United States is currently limited to text messaging, forcing communication in English as opposed to American Sign Language (ASL), the preferred language. Because ASL is a visual language, mobile video phones have the potential to give Deaf people access to real-time mobile communication in their preferred language. However, even today's best video compression techniques can not yield intelligible ASL at limited cell phone network bandwidths. Motivated by this constraint, we conducted one focus group and two user studies with members of the Deaf Community to determine the intelligibility effects of video compression techniques that exploit the visual nature of sign language. Inspired by eye tracking results that show high resolution foveal vision is maintained around the face, we studied region-of interest encodings (where the face is encoded at higher quality) as well as reduced frame rates (where fewer, better quality, frames are displayed every second). At all bit rates studied here, participants preferred moderate quality increases in the face region, sacrificing quality in other regions. They also preferred slightly lower frame rates because they yield better quality frames for a fixed bit rate. The limited processing power of cell phones is a serious concern because a real-time video encoder and decoder will be needed. Choosing less complex settings for the encoder can reduce encoding time, but will affect video quality. We studied the intelligibility effects of this tradeoff and found that we can significantly speed up encoding time without severely affecting intelligibility. These results show promise for real-time access to the current low-bandwidth cell phone network through sign-language-specific encoding techniques. PMID- 18416522 TI - Protection against aflatoxin B1-induced cytotoxicity by expression of the cloned aflatoxin B1-aldehyde reductases rat AKR7A1 and human AKR7A3. AB - The reduction of the aflatoxin B 1 (AFB 1) dialdehyde metabolite to its corresponding mono and dialcohols, catalyzed by aflatoxin B 1-aldehyde reductase (AFAR, rat AKR7A1, and human AKR7A3), is greatly increased in livers of rats treated with numerous chemoprotective agents. Recombinant human AKR7A3 has been shown to reduce the AFB 1-dialdehyde at rates greater than those of the rat AKR7A1. The activity of AKR7A1 or AKR7A3 may detoxify the AFB 1-dialdehyde, which reacts with proteins, and thereby inhibits AFB 1-induced toxicity; however, direct experimental evidence of this hypothesis was lacking. Two human B lymphoblastoid cell lines, designated pMF6/1A2/AKR7A1 and pMF6/1A2, were genetically engineered to stably express AKR7A1 and/or cytochrome P4501A2 (1A2). The pMF6/1A2/AKR7A1 cells were refractory to the cytotoxic effects of 3 ng/mL AFB 1, in comparison to pM6/1A2 cells, which were more sensitive. Diminished protection occurred at higher concentrations of AFB 1 in pMF6/1A2/AKR7A1 cells, suggesting that additional factors were influencing cell survival. COS-7 cells were transfected with either vector control, rat AKR7A1, or human AKR7A3, and the cells were treated with AFB 1-dialdehyde. There was a 6-fold increase in the dialdehyde LC 50, from 66 microM in vector-transfected cells to 400 microM in AKR7A1-transfected cells, and an 8.5-fold increase from 35 microM in vector transfected cells to 300 microM in AKR7A3-transfected cells. In both cases, this protective effect of the AFAR enzyme was accompanied by a marked decrease in protein adducts. Fractionation of the cellular protein showed that the mitochondria/nuclei and microsomal fractions contained the highest concentration of protein adducts. The levels of human AKR7A3 and AKR7A2 were measured in 12 human liver samples. The expression of AKR7A3 was detectable in all livers and lower than those of AKR7A2 in 11 of the 12 samples. Overall, these results provide the first direct evidence of a role for rat AKR7A1 and human AKR7A3 in protection against AFB 1-induced cytotoxicity and protein adduct formation. PMID- 18416524 TI - Theoretical and experimental study on a self-assembling polysaccharide forming nanochannels: static and dynamic effects induced by a soft confinement. AB - It is well-known that the polysaccharide scleroglucan (Sclg) exhibits a triple helix conformation (triplex) and it is able to form hydrogels in water solution. Furthermore, these hydrogels are influenced by the presence of borax, in terms of rheological and drug release properties. In previous works, we showed that the presence of borax stabilizes the intertriplex interactions and that the property variations, induced by borax, can be fully explained, considering that the Sclg triplexes can form nanochannel-like structures. In this paper, the stability of these aggregates has been experimentally studied by means of atomic force microscopy (AFM) and theoretically investigated by means of molecular dynamics (MD) simulations. The simulations indicate that the borax stabilizes nanochannel like structures when seven triplexes are considered. The simultaneous presence of different Sclg triplexes in a narrow space strongly influences the properties of confined water molecules in a way similar, in many aspects, to that of water molecules located in the inner part of well-defined nanochannels (e.g., diffusion inside carbon nanotubes). As a consequence, also the conformational properties of flanking regions of Sclg triplexes are influenced. Furthermore, differential scanning calorimetry (DSC) data show that the well-known conformational transition occurring at 280 K for Sclg does not take place in the presence of borax. The MD simulations suggest that such lack of transition is a direct consequence of the presence of borax. The role of Na+ counterions in the hydrogel structure is also investigated. PMID- 18416525 TI - Synthesis and structure of a new heptaborate oxoanion isomer: B7O9(OH)5(2-). AB - The nonmetal borate [H3N(CH2)7NH3][B7O9(OH)5].H2O (1) was prepared via a neat reaction of 1,7-diaminoheptane with excess boric acid under mild nonhydrothermal conditions. Single-crystal X-ray characterization of 1 revealed a new isomer of the heptaborate anion, B7O9(OH)5(2-). The heptaborate anion in 1 is comprised of four fused (BO)3 rings sharing three four-coordinate boron atoms and a single three-coordinate oxygen atom. This anion is a structural isomer of another recently described heptaborate anion, consisting of a chain of three (BO)3 rings. Compound 1 crystallized in the triclinic space group P1 with a=9.3266(17) A, b=10.1222(19) A, c=10.847(2) A, alpha=89.422(2) degrees, beta=82.349(2) degrees, gamma=75.158(2) degrees, V=980.7(3) A3, and Z=2. PMID- 18416526 TI - Synthesis and characterization of (3,1) Ru2(F3ap)4(NCS) and (3,1) Ru2(F3ap)3(F2Oap)(NCS) where F3ap is the 2-(2,4,6-trifluoroanilino)pyridinate anion. AB - Two isothiocyanate diruthenium complexes, (3,1) Ru2(F3ap)4(NCS) 1 and (3,1) Ru2(F3ap)3(F2Oap)(NCS)2 (where F3ap=2,4,6-trifluoroanilinopyridinate anion), were synthesized from (3,1) Ru2(F3ap)4Cl and SCN(-) under different experimental conditions. Each compound was examined as to its structural, electrochemical, spectroscopic, and magnetic properties. Compound 1 contains three unpaired electrons as its parent compound but 2 is diamagnetic. The X-ray molecular structures of 1 and 2 reveal that the NCS group is coordinated to the dimetal unit via nitrogen in both compounds with the Ru-N-C bond angle being 176.5 degrees for 1 and 166.0 degrees for 2. An elongation of the Ru-Ru bond distance and a shortening of both the Ru-Np (p=pyridyl) and the Ru-Na (a=anilino) bond lengths is seen upon going from (3,1) Ru2(F3ap)4Cl to 2, but the conversion of (3,1) Ru2(F3ap)4Cl to 1 does not affect significantly structural features of the Ru2(L) 4 framework. Compound 1 undergoes one reduction and two oxidations, all three of which involve the dimetal core, whereas 2 undergoes two metal-centered reductions, one metal-centered oxidation, and one ligand-based oxidation due to the presence of the F2Oap ligand on the Ru2 complex. The reactivity of 1 with SCN(-) was also investigated. PMID- 18416527 TI - Toluene-sandwiched trinuclear copper(I) and silver(I) triazolates and phosphine adducts of dinuclear copper(I) and silver(I) triazolates. AB - Cu (I) and Ag (I) complexes of the fluorinated triazolate ligand [3,5-(C3F7)2Tz]( ) have been synthesized using the corresponding metal(I) oxides and the triazole. They form pi-acid/base adducts with toluene, leading to [Tol][M3][Tol] ([Tol]=toluene; [M3]={[3,5-(C3F7)2Tz]Cu}3 or {[3,5-(C3F7)2Tz]Ag}3) type structures. Packing diagrams show the presence of extended chains of the type {[Tol][M3][Tol]}infinity, but the intertoluene ring distances are too long for significant pi-arene/pi-arene contacts. These copper and silver triazolates react with PPh3 (at a 1:1 metal ion/P molar ratio), leading to dinuclear {[3,5 (C3F7)2Tz]Cu(PPh3)}2 and {[3,5-(C3F7) 2Tz]Ag(PPh3)}2. They feature a six-membered Cu(mu-N-N) 2Cu or Ag(mu-N-N)2Ag core with a boat conformation. PMID- 18416528 TI - Organotitanoxanes with unique structure among transition-element organometallic oxide derivatives. AB - The synthesis of novel titanoxane compounds, [{(TiCl)(Ti)[mu-(eta(5)-C5Me4SiMe2O kappaO)]2(mu-O)}2(mu-O)] (4) and [{Ti[mu-(eta(5)-C5Me4SiMe2O-kappaO)](mu-O)}6] (5), by controlled hydrolysis of a dinuclear titanium/oxo complex is described. Complexes 4 and 5 show unprecedented structural features for organometallic oxide derivatives of transition elements and represent unique fully characterized examples of tetra- and hexanuclear organo-transition-metal oxide compounds with an open-chain and a monocyclic structure, respectively. PMID- 18416529 TI - Lithium-palladium complex supported by phosphonatophosphine and chloride ligands. AB - The reaction of the phosphonatophosphine ligand Ph2PCH(SiMe3)P(O)(OEt)2 with [PdCl2(COD)] in the presence of LiCl afforded the neutral, mixed-metal complex [LiPd2CI5{mu-Ph2PCH2P(O)(OEt)2}2] in which the Li(+) cation is tetrahedrally coordinated by the PO bonds and two Pd-bound Cl atoms. PMID- 18416530 TI - Peptides are building blocks of heat-induced fibrillar protein aggregates of beta lactoglobulin formed at pH 2. AB - The proteinaceous material present in beta-lactoglobulin fibrils formed after heating (20 h at 85 degrees C) at pH 2 was identified during this study. Fibrils were separated from the nonaggregated material, and the fibrils were dissociated using 8 M guanidine chloride and 0.1 M 1,4-dithiothreitol (pH 8). Characterization of the different fractions was performed using thioflavin T fluorescence, high-performance size-exclusion chromatography, reversed-phase HPLC, and mass spectrometry (MALDI-TOF). Beta-lactoglobulin was found to be hydrolyzed into peptides with molecular masses between 2000 and 8000 Da, and the fibrils were composed of a part of these peptides and not intact beta lactoglobulin. The majority of the peptides (both aggregated and nonaggregated) were a result from cleavage of the peptide bonds before or after aspartic acid residues. Explanations for the presence of certain peptide fragments in the fibrils are the hydrophobicity, low charge, charge distribution, and capacity to form beta-sheets. PMID- 18416531 TI - CrCl2-promoted stereospecific and stereoselective alkyl- and silylcyclopropanation of alpha,beta-unsaturated amides. AB - An efficient chromium-promoted alkyl- or silylcyclopropanation of alpha,beta unsaturated amides is described. These reactions can be carried out on (E)- or (Z)-alpha,beta-enamides in which the C-C double bond is di-, or trisubstituted. This process takes place with total stereospecificity and the new stereogenic center is generated with high or total stereoselectivity. Some synthetic applications of the obtained silylcyclopropyl amides are also reported. Two mechanisms based on the generation of carbenoid or carbene complexes have been proposed to explain this cyclopropanation reaction. PMID- 18416532 TI - Cyclic nitriles: stereodivergent addition-alkylation-cyclization to cis- and trans-abietanes. AB - Diverse cyclic hydroxy nitriles are readily synthesized through sequential 1,2 1,4-Grignard addition-methylations to 3-oxo-1-cyclohexene-1-carbonitrile. Acid catalyzed intramolecular cyclizations of the cyclic hydroxy nitriles reveal fundamental stereoselectivity trends in Friedel-Crafts cyclizations to cis- and trans-abietanes. In contrast to previous assumptions, comparative cationic cyclizations with electron-rich and electron-poor aromatic nucleophiles exhibit similar preferences for cyclization to cis-abietanes. Optimizing the cyclizations for trans-abietanes has identified ZrCl 4 as an exceptional Lewis acid which, for cyclizations of iminolactones, favors trans-abietanes as the only observable diastereomer. The sequential oxonitrile addition-Friedel-Crafts cyclization strategy provides a rapid, stereodivergent synthesis of cis- or trans-abietanes, demonstrates the dramatic influence of ZrCl 4 in promoting cationic cyclizations, and in contrast to previous assumptions suggests that the cyclization stereoselectivity is not correlated with the electronic nature of the aromatic nucleus. PMID- 18416533 TI - Atmospheric pressure air direct current glow discharge ionization source for ion mobility spectrometry. AB - A new atmospheric pressure air direct current glow discharge (DCGD) ionization source has been developed for ion mobility spectrometry (IMS) to overcome the regularity problems associated with the conventional (63)Ni source and the instability of the negative corona discharge. Its general electrical characteristics were experimentally investigated. By equipping it to IMS, a higher sensitivity was obtained compared to that of a (63)Ni source and corona discharge, and a linear dynamic range from 20 ppb to 20 ppm was obtained for m xylene. Primary investigations showed that alkanes, such as pentane, which are nondetectable or insensitively detectable with (63)Ni-IMS, can be efficiently detected by DCGD-IMS and the detection limit of 10 ppb can be reached. The preliminary results have shown that the new DCGD ionization source has great potential applications in IMS, such as online monitoring of environment pollutants and halogenated compounds. PMID- 18416534 TI - Flame etching enhances the sensitivity of carbon-fiber microelectrodes. AB - Small sensors are useful for in vivo measurements and probing small spaces. In this paper, we compare two methods of fabrication of small, cylindrical carbon fiber microelectrodes: flame-etching and electrochemical etching. With both methods, microelectrodes can be fabricated with tip diameters of 1 to 3 microm. Electrodes were tested with fast-scan cyclic voltammetry. Flame etching resulted in electrodes that have larger S/N ratios and higher currents per unit area for 1 microM dopamine than normal carbon-fiber microelectrodes or electrochemically etched electrodes. Therefore, the increased sensitivity is not just a property of size. The flame-etched surfaces had nanometer-scale surface features that were not observed on the other electrodes and exhibited increased sensitivity for other electroactive compounds found in the brain, including ascorbic acid, DOPAC, and serotonin. Faster kinetics and a faster response to a step change in dopamine were also observed, when the applied waveform was -0.4 to 1.0 V and back at 400 V/s. The sensitivity of the flame-etched electrodes was enhanced by overoxidizing the surface. The flame-etched electrodes were used to detect dopamine release in anesthetized rats after a single stimulation pulse. The small flame-etched electrodes will facilitate measurements of low concentrations in discrete brain regions or small organisms. PMID- 18416535 TI - Membrane features and activity of GPI-anchored enzymes: alkaline phosphatase reconstituted in model membranes. AB - The influence of membrane lipid environment on the activity of GPI-anchored enzymes was investigated with human placental alkaline phosphatase reconstituted by a detergent-dialysis technique in liposomes composed of palmitoyloleoylphosphatidylcholine, alone or in mixture with lipids enriched along with the protein within lipid rafts: cholesterol, sphingomyelin, and GM1 ganglioside. The highest V max was recorded for a phosphatidylcholine/10% GM1 mixture (143 +/- 5 nmol of substrate hydrolyzed per minute per microgram of protein), while the lowest for a phosphatidylcholine/30% cholesterol mixture and for raft-mimicking 1:1:1 phosphatidylcholine/sphingolipid/cholesterol liposomes (M:M:M) (57 +/- 3 and 52 +/- 3, respectively). No significant differences in K m were detected. The protein segregation, assessed using the chemical cross-linker bis(sulfosuccinimidyl)suberate, increased with the protein:lipid ratio, within the 1:1200-1:4800 protein:lipid molar ratio range, but did not affect enzyme activity. The activity decreased when the order of the lipid bilayers was increased, higher for those containing cholesterol, as judged by steady-state fluorescence polarization of 1,6-diphenyl-1,3,5-hexatriene. Finally, the GPI enzyme activity was affected by membrane curvature. This result was suggested by a strong inverse correlation (Pearson's correlation coefficient = 0.91; p < 0.0001) between activity and liposome diameter, measured by laser light scattering and ranging between 59 +/- 6 nm for a phosphatidylcholine/10% GM1 mixture (displaying the highest activity) and 188 +/- 25 nm for a phosphatidylcholine/30% cholesterol mixture and 185 +/- 23 nm for raft-mimicking liposomes (displaying the lowest activities). The activity-membrane curvature relationship was further confirmed by comparing the activity of proteoliposomes having different sizes but identical lipid compositions. These data open the possibility that the activity of GPI-anchored enzymes may be modulated by membrane microenvironment features, in particular by membrane curvature and cholesterol-enriched ordered microenvironments, such as those of lipid rafts. PMID- 18416536 TI - Crystal structure of the pyocyanin biosynthetic protein PhzS. AB - The human pathogen Pseudomonas aeruginosa produces pyocyanin, a blue-pigmented phenazine derivative, which is known to play a role in virulence. Pyocyanin is produced from chorismic acid via the phenazine pathway, nine proteins encoded by a gene cluster. Phenazine-1-carboxylic acid, the initial phenazine formed, is converted to pyocyanin in two steps that are catalyzed by the enzymes PhzM and PhzS. PhzM is an adenosylmethionine dependent methyltransferase, and PhzS is a flavin dependent hydroxylase. It has been shown that PhzM is only active in the physical presence of PhzS, suggesting that a protein-protein interaction is involved in pyocyanin formation. Such a complex would prevent the release of 5 methyl-phenazine-1-carboxylate, the putative intermediate, and an apparently unstable compound. Here, we describe the three-dimensional structure of PhzS, solved by single anomalous dispersion, at a resolution of 2.4 A. The structure reveals that PhzS is a member of the family of aromatic hydroxylases characterized by p-hydroxybenzoate hydroxylase. The flavin cofactor of PhzS is in the solvent exposed out orientation typically seen in unliganded aromatic hydroxylases. The PhzS flavin, however, appears to be held in a strained conformation by a combination of stacking interactions and hydrogen bonds. The structure suggests that access to the active site is gained via a tunnel on the opposite side of the protein from where the flavin is exposed. The C-terminal 23 residues are disordered as no electron density is present for these atoms. The probable location of the C-terminus, near the substrate access tunnel, suggests that it may be involved in substrate binding as has been shown for another structural homologue, RebC. This region also may be an element of a PhzM-PhzS interface. Aromatic hydroxylases have been shown to catalyze electrophilic substitution reactions on activated substrates. The putative PhzS substrate, however, is electron deficient and unlikely to act as a nucleophile, suggesting that PhzS may use a different mechanism than its structural relatives. PMID- 18416537 TI - Enantioselectivity in zebrafish embryo toxicity of the insecticide acetofenate. AB - Enantioselectivity in separation and toxicology of chiral xenobiotics have become one of the frontier topics interfacing chemistry and toxicology. In this study, enantiomers of insecticide acetofenate (AF) were separated on selected chiral columns by HPLC, and enantioselectivity in developmental toxicity was evaluated using the zebrafish embryo-larval assays. The AF enantiomers were baseline separated on Chiralcel OD, Chiralpak AD, and Sumichiral OA-2500I columns under optimized conditions. Pure enantiomers were obtained on Chiralcel OD. Optical rotatory dispersion (ORD) and circular dichroism (CD) detectors were used to determine the elution order and CD spectra of the enantiomers. The absolute configuration of enantiomers was identified as S-(+)-AF and R-(-)-AF by the octant rule from force-field calculations and CD spectra. The individual enantiomers were used in 4-day zebrafish embryo-larval bioassays, and a series of developmental end points were measured and compared. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to investigate the expressions of estrogen receptor alpha (ERalpha) in zebrafish embryo exposed to varying enantiomers. While the enantiomers showed no difference in acute toxicity, significant enantioselectivity was observed in developmental toxicities such as yolk sac edema and pericardial edema. The data of qRT-PCR showed that there was about 3.2-fold induction in the mRNA levels of ERalpha between fish exposed to (+)-AF and (-)-AF. The results suggest that enantioselectivity may occur at the developmental level even in the absence of selective acute toxicity and should be considered when evaluating ecotoxicological effects of chiral contaminants. PMID- 18416538 TI - Conformational properties of equilenin-DNA adducts: stereoisomer and base effects. AB - Equilin and equilenin, components of the hormone replacement therapy drug Premarin, can be metabolized to the catechol 4-hydroxyequilenin (4-OHEN). The quinoids produced by 4-OHEN oxidation react with dC, dA, and dG to form unusual stable cyclic adducts, which have been found in human breast tumor tissue. Four stereoisomeric adducts have been identified for each base. These 12 Premarin derived adducts provide a unique opportunity for analyzing effects of stereochemistry and base damage on DNA structure and consequently its function. Our computational studies have shown that these adducts, with obstructed Watson Crick hydrogen-bond edges and near-perpendicular ring systems, have limited conformational flexibility and near-mirror-image conformations in stereoisomer pairs. The dC and dA adducts can adopt major- and minor-groove positions in the double helix, but the dG adducts are positioned only in the major groove. In all cases, opposite orientations of the equilenin rings with respect to the 5' --> 3' direction of the damaged strand are found in stereoisomer pairs derived from the same base, and no Watson-Crick pairing is possible. However, detailed structural properties in DNA duplexes are distinct for each stereoisomer of each damaged base. These differences may underlie observed differential stereoisomer and base dependent mutagenicities and repair susceptibilities of these adducts. PMID- 18416539 TI - Measurement of protein sulfhydryls in response to cellular oxidative stress using gel electrophoresis and multiplexed fluorescent imaging analysis. AB - The significance of free radicals in biology has been established by numerous investigations spanning a period of over 40 years. Whereas there are many intracellular targets for these radical species, the importance of cysteine thiol posttranslational modification has received considerable attention. The current studies present a highly sensitive method for measurement of the posttranslational modification of protein thiols. This method is based on labeling of proteins with monofunctional maleimide dyes followed by 2D gel electrophoresis to separate proteins and multiplexed fluorescent imaging analysis. The method correctly interrogates the thiol/disulfide ratio present in commercially available proteins. Exposure of pulmonary airway epithelial cells to high concentrations of menadione or t-butyl hydroperoxide resulted in the modification of cysteines in more than 141 proteins of which 60 were subsequently identified by MALDI-TOF/TOF MS. Although some proteins were modified similarly by these two oxidants, several showed detectably different maleimide ratios in response to these two agents. Proteins that were modified by one or both oxidants include those involved in transcription, protein synthesis and folding, and cell death/growth. In conclusion, these studies provide a novel procedure for measuring the redox status of cysteine thiols on individual proteins with a clearly demonstrated applicability to interactions of chemicals with pulmonary epithelial cells. PMID- 18416540 TI - Catalytic motors for transport of colloidal cargo. AB - Autonomous micro- and nanomotors should, in principle, deliver materials in a site-directed fashion, powering the assembly of dynamic, nonequilibrium superstructures. Here we demonstrate that catalytic Pt-Au nanomotors can transport a prototypical cargo: polystyrene microspheres. In addition, motors with Ni segments can overcome both Brownian orientational fluctuations and biased rotation of the rod-sphere doublet to enable persistent steerable uniaxial motion in an external magnetic field. Assuming a cargo-independent motive force, the speeds are inversely proportional to the Stokes resistance, which we compute using a completed double-layer boundary integral equation. In addition, we demonstrate motors transporting cargo via chemotaxis toward a H2O2 fuel source. PMID- 18416541 TI - Nanophotonic switch: gold-in-Ga2O3 peapod nanowires. AB - A novel metal-insulator heterostructure made of twinned Ga2O3 nanowires embedding discrete gold particles along the twin boundary was formed through a reaction between gold, gallium, and silica at 800 degrees C during simple thermal annealing. The Au-in-Ga2O3 peapods spontaneously crystallized under phase separation induced by the formation of twin boundaries. The nanostructures were analyzed by field emission scanning (FESEM) and transmission electron microscopes (FETEM), and their photoresponse was investigated using a double-frequency Nd:YAG laser with a wavelength of 532 nm on a designed single-nanowire device. The surface plasmon resonance (SPR) effects of embedded Au nanoparticles are proposed to be responsible for the remarkable photoresponse of these novel structures. PMID- 18416542 TI - Theoretical study of 5-aminolevulinic acid tautomerization: a novel self catalyzed mechanism. AB - 5-Aminolevulinic acid (5ALA) is the key synthetic building block in protoporphyrin IX (PpIX), the heme chromophore in mitochondria. In this study density functional theory calculations were performed on the tautomers of 5ALA and the tautomerization reaction mechanism from its enolic forms (5-amino-4 hydroxypent-3-enoic acid and 5-amino-4-hydroxypent-4-enoic acid) to the more stable 5ALA. The hydrated form 5-amino-4,4-dihydroxypentanoic acid was also studied. The lowest energy pathway of 5ALA tautomerization is by means of autocatalysis, in that an oxygen of the carboxylic group transfers the hydrogen atom as a "crane", with an activation energy of approximately 15 kcal/mol. This should be compared to the barriers of about 35 kcal/mol for water assisted tautomerization, and 60 kcal/mol for direct hydrogen transfer. For hydration of 5ALA, the water catalyzed activation barrier is found to be approximately 35 kcal/mol, approximately 5 kcal/mol lower than direct hydration. PMID- 18416543 TI - Aza-peptidyl Michael acceptors. A new class of potent and selective inhibitors of asparaginyl endopeptidases (legumains) from evolutionarily diverse pathogens. AB - Aza-peptide Michael acceptors with the general structure of Cbz-Ala-Ala-AAsn- trans-CH=CHCOR are a new class of inhibitors specific for the asparaginyl endopeptidases (AE) (legumains). Structure-activity relationships (SARs) were characterized for a set of 31 aza-peptide Michael acceptors with AEs derived from three medically important parasites: the protist Trichomonas vaginalis, the hard tick Ixodes ricinus, and the flatworm Schistosoma mansoni. Despite arising from phylogenetically disparate organisms, all three AEs shared a remarkably similar SAR with lowest IC50 values extending into the picomolar range. The results suggest an evolutionary constraint on the topography of the prime side of the active site. SAR also revealed that esters in the P1' position are more potent than disubstituted amides and that monosubstituted amides and alkyl derivatives show little or no inhibition. The preferred P1' residues have aromatic substituents. Aza-asparaginyl Michael acceptors react with thiols, which provides insight into the mechanism of their inhibition of asparaginyl endopeptidases. PMID- 18416545 TI - Application of belief theory to similarity data fusion for use in analog searching and lead hopping. AB - A wide variety of computational algorithms have been developed that strive to capture the chemical similarity between two compounds for use in virtual screening and lead discovery. One limitation of such approaches is that, while a returned similarity value reflects the perceived degree of relatedness between any two compounds, there is no direct correlation between this value and the expectation or confidence that any two molecules will in fact be equally active. A lack of a common framework for interpretation of similarity measures also confounds the reliable fusion of information from different algorithms. Here, we present a probabilistic framework for interpreting similarity measures that directly correlates the similarity value to a quantitative expectation that two molecules will in fact be equipotent. The approach is based on extensive benchmarking of 10 different similarity methods (MACCS keys, Daylight fingerprints, maximum common subgraphs, rapid overlay of chemical structures (ROCS) shape similarity, and six connectivity-based fingerprints) against a database of more than 150,000 compounds with activity data against 23 protein targets. Given this unified and probabilistic framework for interpreting chemical similarity, principles derived from decision theory can then be applied to combine the evidence from different similarity measures in such a way that both capitalizes on the strengths of the individual approaches and maintains a quantitative estimate of the likelihood that any two molecules will exhibit similar biological activity. PMID- 18416544 TI - In vitro and in vivo evaluation of bifunctional bisthiosemicarbazone 64Cu complexes for the positron emission tomography imaging of hypoxia. AB - The copper(II) bisthiosemicarbazonato complex, copper-diacetyl-bis(N4 methylthiosemicarbazonate) (Cu-ATSM), has been used clinically as a positron emission tomography (PET) tracer for the delineation of hypoxia. Six novel, asymmetric bis(thiosemicarbazones) derived from diacetyl-2-(4-N-methyl-3 thiosemicarbazone)-3-(4-N-amino-3-thiosemicarbazone) (H2ATSM/A), one of which contained a nitroimidazole functionality, were radiolabeled with 64Cu (t1/2=12.7 h, beta+=19.3%). In vitro studies were performed on three of the compounds using EMT6 mammary carcinoma cells under hypoxic and normoxic conditions. All compounds displayed rapid cellular association and appreciable hypoxic selectivity with increased uptake under normoxic and hypoxic conditions when compared to 64Cu ATSM. Biodistribution and small animal PET imaging studies were then carried out in vivo using two compounds in EMT6 tumor-bearing mice. The compounds showed high tumor uptake, but also substantial accumulation in the liver. These complexes demonstrate that H 2ATSM/A represents a novel and versatile synthetic platform that can be utilized to provide hypoxic cell selectivity through functionalization of the bisthiosemicarbazonate group. PMID- 18416546 TI - 3D metal-organic frameworks based on elongated tetracarboxylate building blocks for hydrogen storage. AB - Two 3D metal-organic frameworks (MOFs) with a new biphenol-derived tetracarboxylate linker and Cu(II) and Zn(II) metal-connecting points were synthesized and characterized by single-crystal X-ray crystallographic studies. The two isostructural MOFs exhibit distorted PtS network topology and show markedly different framework stability. The porosity and hydrogen uptake of the frameworks were determined by gas adsorption experiments. PMID- 18416547 TI - New attempt at directly generating superbasicity on mesoporous silica SBA-15. AB - Direct generation of superbasicity on mesoporous silica SBA-15 was realized by tailoring the host-guest interaction, and calcium species were selected as the guest in modifying SBA-15. The results show that calcium species could be homogeneously distributed on the surface of SBA-15. Because of the host-guest interaction, the decomposition of the supported calcium nitrate was apparently easier than the bulk one. Surprisingly, the calcium nitrates modified SBA-15 (CaNS) samples exhibited superbasicity with good preservation of the mesostructure after activation, differing from the potassium nitrate loaded SBA 15 samples that displayed weak basicity with collapsed mesostructure. The present superbasic CaNS materials also possess good water resistance and high surface areas, up to 429 m(2) g(-1), which is promising for their potential applications in adsorption and catalysis. Further investigation concerning the roles played by the guest in basicity formation on SBA-15 was conducted. The samples modified by Group 2 metal nitrates showed strong basicity with base strength (H-) of 22.5 27.0 and good preservation of mesostructure. In contrast, loading Group 1 metal nitrates on SBA-15 produced samples with weak basicity ( H-=9.3-15.0) and collapsed mesostructure after activation. Such differences can be related to the interaction between the resulting metal oxide and the silica support, as well as the mobility of the cations in the metal oxide. PMID- 18416548 TI - Nanosecond CO photodissociation and excited-state character of [Ru(X)(X')(CO)2(N,N'-diisopropyl-1,4-diazabutadiene)] (X=X'=Cl or I; X=Me, X'=I; X=SnPh3, X'=Cl) studied by time-resolved infrared spectroscopy and DFT calculations. AB - The character and dynamics of the low-lying excited states of [Ru(X)(X')(CO)2(iPr dab)] (X=X'=Cl or I; X=Me, X'=I; X=SnPh3, X'=Cl; iPr-dab=N, N'-diisopropyl-1,4 diazabutadiene) were studied experimentally by pico- and nanosecond time-resolved IR spectroscopy (TRIR) and (for X=X'=Cl or I) computationally using density functional theory (DFT) and time-dependent DFT (TD-DFT) techniques. The lowest allowed electronic transition occurs between 390 and 460 nm and involves charge transfer from the Ru(halide)(CO) 2 unit to iPr-dab, denoted (1)MLCT/XLCT (metal to-ligand/halide-to-ligand charge transfer). The lowest triplet state is well modeled by UKS-DFT-CPCM calculations, which quite accurately reproduce the excited-state IR spectrum in the nu(CO) region. It has a (3)MLCT/XLCT character with an intraligand (iPr-dab) (3)pipi* admixture. TRIR spectra of the lowest triplet excited state show two nu(CO) bands that are shifted to higher energies from their corresponding ground-state positions. The magnitude of this upward shift increases as a function of the ligands X and X' [(I)2 < (Sn)(Cl) < (Me)(I) < (Cl)2] and reveals increasing contribution of the Ru(CO)2-->dab MLCT character to the excited state. The lowest triplet state of [Ru(Cl)2(CO)2(iPr-dab)] undergoes a approximately 10 ps relaxation that is followed by CO dissociation, producing cis(CO,CH 3CN),trans(Cl,Cl)-[Ru(Cl)2(CH 3CN)(CO)(iPr-dab)] with a unity quantum yield and 7.2 ns lifetime and without any observable intermediate. To our knowledge, this is the first example of a "slow" CO dissociation from a thermally equilibrated triplet charge-transfer excited state. PMID- 18416549 TI - Three new Cu-azido polymers and their systematic interconversion: role of the amount of the blocking amine on the structural diversity and magnetic behavior. AB - Three new coordination polymers [Cu5(N3)10(en)2]n (1), [Cu6(N3)12(en)4]n (2), and [Cu4(N3)8(en)4]n (3) have been synthesized in a controlled manner by treatment of a 1:2 mixture of Cu(NO3)2 and NaN3 with varying amount of ethylenediamine (en). Single-crystal structure analyses clearly indicated that the puckered Cu4 biscubane unit in 1 gradually opens to a slightly more open Cu4 macrocyclic unit in 2 when more en approaches to the Cu4 core. Upon addition of further en, an open Cu4 linear secondary building unit was obtained in complex 3. Complex 1 contains four different kinds of bridging modes of the azide anion and is a complicated 3D polymer. Similarly, complexes 2 and 3 are 3D and 2D polymers, respectively, containing three different kinds of bridging azides. Complex 3 contains two very rare cis end-to-end (EE) and single-end-on (EO) azido modes. Structural transformation from 1 to 3 was monitored and explained qualitatively. Variable-temperature magnetic studies in the temperature range of 300-2 K reveal the existence of dominant ferromagnetic behavior in all the three cases with a metamagnetic-type behavior in complex 1 with the critical field of transition at 0.8 T. The purity of all the complexes were established by elemental analyses, as well as by the powder XRD patterns that matched well with the expected patterns from the single-crystal structure analysis. PMID- 18416550 TI - Theoretical study of pyrazolate-bridged dinuclear platinum(II) complexes: interesting potential energy curve of the lowest energy triplet excited state and phosphorescence spectra. AB - Four kinds of 3,5-dialkylpyrazolate(R2pz)-bridged dinuclear platinum(II) complexes [Pt2(mu-R2pz)2(dfppy)2] (dfppy=2-(2,4-difluorophenyl)pyridine; R2pz=pyrazolate in 1, 3,5-dimethylpyrazolate in 2, 3-methyl-5- tert butylpyrazolate in 3, and 3,5-bis(tert-butyl)pyrazolate in 4) were theoretically investigated by the DFT(B3PW91) method. The Stokes shift of their phosphorescence spectra was discussed on the basis of the potential energy curve (PEC) of the lowest energy triplet excited state (T1). This PEC significantly depends on the bulkiness of substituents on pz. In 1 and 2, bearing small substituents on pz, one local minimum is present in the T1 state besides a global minimum. The local minimum geometry is similar to the S0-equilibrium one. The T1 state at this local minimum is characterized as the pi-pi* excited state in dfppy, where the dpi orbital of Pt participates in this excited state through an antibonding interaction with the pi orbital of dfppy; in other words, this triplet excited state is assigned as the mixture of the ligand-centered pi-pi* excited and metal to-ligand charge transfer excited state ((3)LC/MLCT). The geometry of the T1 global minimum is considerably different from the S0-equilibrium one. The T1 state at the global minimum is characterized as the triplet metal-metal-to-ligand charge transfer ((3)MMLCT) excited state, which is formed by the one-electron excitation from the dsigma-dsigma antibonding orbital to the pi* orbital of dfppy. Because of the presence of the local minimum, the geometry change in the T1 state is suppressed in polystyrene at room temperature (RT) and frozen 2 methyltetrahydrofuran (2-MeTHF) at 77 K. As a result, the energy of phosphorescence is almost the same in these solvents. In fluid 2-MeTHF at RT, on the other hand, the geometry of the T1 state easily reaches the T1-global minimum. Because the T1-global minimum geometry is considerably different from the S0-equilibrium one, the phosphorescence occurs at considerably low energy. These are the reasons why the Stokes shift is very large in fluid 2-MeTHF but small in polystyrene and frozen 2-MeTHF. In 3 and 4, bearing bulky tert-butyl substituents on pz, only the T1-global minimum is present but the local minimum is not. The electronic structure of this T1-global minimum is assigned as the (3)MMLCT excited state like 1 and 2. Though frozen 2-MeTHF suppresses the geometry change of 3 and 4 in the T1 state, their geometries moderately change in polystyrene because of the absence of the T1-local minimum. As a result, the energy of phosphorescence is moderately lower in polystyrene than in frozen 2 MeTHF. The T1-global minimum geometry is much different from the S0-equilibrium one in 3 but moderately different in 4, which is interpreted in terms of the symmetries of these complexes and the steric repulsion between the tert-butyl group on pz and dfppy. Thus, the energy of phosphorescence of 3 is much lower in fluid 2-MeTHF than in frozen 2-MeTHF like 1 and 2 but that of 4 is moderately lower; in other words, the Stokes shift in fluid 2-MeTHF is small only in 4. PMID- 18416551 TI - Impact of the hard-to-cook phenomenon on phenolic antioxidants in dry beans (Phaseolus vulgaris). AB - Epidemiological studies have established a link between consumption of dry beans and lower incidence of degenerative diseases. This relationship is attributed in part to properties of natural antioxidants present in beans. The objective of this study was to determine if the hard-to-cook (HTC) phenomenon in beans had a negative effect on the content of free and bound phenolic antioxidants and antioxidant capacity. Folin-Ciocalteu, Trolox equivalent antioxidant capacity, and HPLC methods were used to quantify the content of phenolic acids and antioxidant capacity. Results showed that the HTC phenomenon did not equally affect the content and antioxidant capacity of phenolic acids in different bean cultivars. Black beans were most affected, the contents of free and acid hydrolyzable phenolic acids being reduced by 35 and 36%, respectively, and the antioxidant activity by 18 and 25%, respectively. This study showed that the HTC phenomenon affected a potential nutritive characteristic of dry beans. PMID- 18416552 TI - Quantification of cysteine S-conjugate of 3-sulfanylhexan-1-ol in must and wine of petite arvine vine by stable isotope dilution analysis. AB - Making use of a convenient synthetic approach to prepare the deuterated S-3 (hexan-1-ol)-cysteine by a Michael addition reaction, an analytical method was developed to measure the presence of the cysteine S-conjugate, precursor of 3 sulfanylhexan-1-ol (3-mercaptohexan-1-ol), in must and wine from Petite Arvine vine. The method uses a stable isotope dilution assay with a suitable one-step sample preparation and HPLC-MS detection. The method has limits of detection and quantification of 3 and 10 microg/L, respectively. A correlation between the increase of the precursor concentration and the increase of the degree of rot has been established. PMID- 18416553 TI - Release of protein, lipid, and vitamin E from almond seeds during digestion. AB - The evaluation of the bioaccessibility of almond nutrients is incomplete. However, it may have implications for the prevention and management of obesity and cardiovascular disease. This study quantified the release of lipid, protein, and vitamin E from almonds during digestion and determined the role played by cell walls in the bioaccessibility of intracellular nutrients. Natural almonds (NA), blanched almonds (BA), finely ground almonds (FG), and defatted finely ground almonds (DG) were digested in vitro under simulated gastric and gastric followed by duodenal conditions. FG were the most digestible with 39, 45, and 44% of lipid, vitamin E, and protein released after duodenal digestion, respectively. Consistent with longer residence time in the gut, preliminary in vivo studies showed higher percentages of nutrient release, and microscopic examination of digested almond tissue demonstrated cell wall swelling. Bioaccessibility is improved by increased residence time in the gut and is regulated by almond cell walls. PMID- 18416554 TI - Acremonium zeae, a protective endophyte of maize, produces dihydroresorcylide and 7-hydroxydihydroresorcylides. AB - Acremonium zeae has been characterized as a protective endophyte of maize and displays antifungal activity against other fungi. Pyrrocidines A and B were discovered to be the metabolites accounting for this activity. During a population survey of A. zeae isolates from maize seeds produced in nine states to determine their ability to produce pyrrocidines, another metabolite of A. zeae, unrelated to the pyrrocidines, was found to have widespread occurrence (105 of 154 isolates) and to be produced in amounts comparable to the pyrrocidines. Further chemical studies of fermentation extracts of an A. zeae isolate (NRRL 45893) from maize led to the identification of a new compound, dihydroresorcylide, the saturated analogue of cis-resorcylide. Also identified were the two diastereomers of 7-hydroxydihydroresorcylide. Dihydroresorcylide and pyrrocidines A and B were detected by LC-APCI-MS in symptomatic maize kernels from ears that were wound-inoculated in the milk stage with A. zeae NRRL 34559. PMID- 18416555 TI - Nondestructive application of laser-induced fluorescence spectroscopy for quantitative analyses of phenolic compounds in strawberry fruits (Fragaria x ananassa). AB - Laser-induced fluorescence spectroscopy (LIFS) was nondestructively applied on strawberries (EX = 337 nm, EM = 400-820 nm) to test the feasibility of quantitatively determining native phenolic compounds in strawberries. Eighteen phenolic compounds were identified in fruit skin by UV and MS spectroscopy and quantitatively determined by use of rp-HPLC for separation and diode-array or chemical reaction detection. Partial least-squares calibration models were built for single phenolic compounds by means of nondestructively recorded fluorescence spectra in the blue-green wavelength range using different data preprocessing methods. The direct orthogonal signal correction resulted in r (2) = 0.99 and rmsep < 8% for p-coumaroyl-glucose, and r (2) = 0.99 and rmsep < 24% for cinnamoyl-glucose. In comparison, the correction of the fluorescence spectral data with simultaneously recorded reflectance spectra did not further improve the calibration models. Results show the potential of LIFS for a rapid and nondestructive assessment of contents of p-coumaroyl-glucose and cinnamoyl glucose in strawberry fruits. PMID- 18416556 TI - Characterization of the p-coumaric acid decarboxylase from Lactobacillus plantarum CECT 748(T). AB - It was previously reported that cell cultures from Lactobacillus plantarum CECT 748 (T) were able to decarboxylate phenolic acids, such as p-coumaric, m coumaric, caffeic, ferulic, gallic, and protocatechuic acid. The p-coumaric acid decarboxylase (PDC) from this strain has been overexpressed and purified. This PDC differs at its C-terminal end when compared to the previously reported PDC from L. plantarum LPCHL2. Because the C-terminal region of PDC is involved in enzymatic activity, especially in substrate activity, it was decided to biochemically characterize the PDC from L. plantarum CECT 748 (T). Contrarily to L. plantarum LPCHL2 PDC, the recombinant PDC from L. plantarum CECT 748 (T) is a heat-labile enzyme, showing optimal activity at 22 degrees C. This PDC is able to decarboxylate exclusively the hydroxycinnamic acids p-coumaric, caffeic, and ferulic acids. Kinetic analysis showed that the enzyme has a 14-fold higher K(M) value for p-coumaric and caffeic acids than for ferulic acid. PDC catalyzes the formation of the corresponding 4-vinyl derivatives (vinylphenol and vinylguaiacol) from p-coumaric and ferulic acids, respectively, which are valuable food additives that have been approved as flavoring agents. The biochemical characteristics showed by L. plantarum PDC should be taken into account for its potential use in the food-processing industry. PMID- 18416557 TI - DNA polymerase I-mediated translesion synthesis in RecA-independent DNA interstrand cross-link repair in E. coli. AB - DNA interstrand cross-links (ICLs) are mainly repaired by the combined action of nucleotide excision repair and homologous recombination in E. coli. Genetic data also suggest the existence of a nucleotide excision repair-dependent, homologous recombination-independent ICL repair pathway. The involvement of translesion synthesis in this pathway has been postulated; however, the molecular mechanism of this pathway is not understood. To examine the role of translesion synthesis in ICL repair, we generated a defined substrate with a single psoralen ICL that mimics a postincision structure generated by nucleotide excision repair. We demonstrated that the Klenow fragment (DNA polymerase I) performs translesion synthesis on this model substrate. This in vitro translesion synthesis assay will help in understanding the basic mechanism of a postincision translesion synthesis process in ICL repair. PMID- 18416558 TI - Optimized proteomic analysis on gels of cell-cell adhering junctional membrane proteins. AB - A high level of structural organization of functional membrane domains in very narrow regions of a plasma membrane is crucial for the functions of plasma membranes and various other cellular functions. Conventional proteomic analyses are based on total soluble cellular proteins. Thus, because of insolubility problems, they have major drawbacks for use in analyses of low-abundance proteins enriched in very limited and specific areas of cells, as well as in analyses of the membrane proteins in two-dimensional gels. We optimized proteomic analyses of cell-cell adhering junctional membrane proteins on gels. First, we increased the purity of cell-cell junctions, which are very limited and specific areas for cell cell adhesion, from hepatic bile canaliculi. We then enriched junctional membrane proteins via a guanidine treatment; these became selectively detectable on two- dimensionally electrophoresed gels after treatment with an extremely high concentration of NP-40. The framework of major junctional integral membrane proteins was shown on gels. These included six novel junctional membrane proteins of type I, type II, and tetraspanin, which were identified by mass spectrometry and by a database sequence homology search, as well as 12 previously identified junctional membrane proteins, such as cadherins and claudins. PMID- 18416559 TI - Overall kinetic mechanism of saccharopine dehydrogenase (L-glutamate forming) from Saccharomyces cerevisiae. AB - Kinetic studies were carried out for histidine-tagged saccharopine reductase from Saccharomyces cerevisiae at pH 7.0, suggesting a sequential mechanism with ordered addition of reduced nicotinamide adenine dinucleotide phosphate (NADPH) to the free enzyme followed by L-alpha-aminoadipate-delta-semialdehyde ( L-AASA) which adds in rapid equilibrium prior to l-glutamate in the forward reaction direction. In the reverse reaction direction, nicotinamide adenine dinucleotide phosphate (NADP) adds to the enzyme followed by addition of saccharopine. Product inhibition by NADP is competitive vs NADPH and noncompetitive vs alpha-AASA and L glutamate, suggesting that the dinucleotide adds to the free enzyme prior to the aldehyde. Saccharopine is noncompetitive vs NADPH, alpha-AASA, and L-glutamate. In the direction of saccharopine oxidation, NADPH is competitive vs NADP and noncompetitive vs saccharopine, L-glutamate is noncompetitive vs both NADP and saccharopine, while L-AASA is noncompetitive vs saccharopine and uncompetitive vs NADP. The sequential mechanism is also corroborated by dead-end inhibition studies using analogues of AASA, L-glutamate, and saccharopine. 2-Amino-6 heptenoic acid was chosen as a dead-end analogue of L-AASA and is competitive vs AASA, uncompetitive vs NADPH, and noncompetitive vs L-glutamate. alpha Ketoglutarate (alpha-Kg) serves as the dead-end analogue of L-glutamate and is competitive vs L-glutamate and uncompetitive vs L-AASA and NADPH. In the direction of saccharopine oxidation, N-oxalylglycine, L-pipecolic acid, L leucine, alpha-ketoglutarate, glyoxylic acid, and L-ornithine were used as dead end analogues of saccharopine and showed competitive inhibition vs saccharopine and uncompetitive inhibition vs NADP. The equilibrium constant for the reaction was measured at pH 7.0 by monitoring the change in absorbance of NADPH and is 200 M(-1). The value is in good agreement with the value determined using the Haldane relationship. PMID- 18416560 TI - Zebrafish neuroglobin is a cell-membrane-penetrating globin. AB - Neuroglobin (Ngb) is a recently discovered vertebrate heme protein that is expressed in the brain and can reversibly bind oxygen. Mammalian Ngb is involved in neuroprotection under oxidative stress conditions, such as ischemia and reperfusion. We previously demonstrated that human ferric Ngb binds to the alpha subunit of heterotrimeric G proteins (Galphai) and acts as a guanine nucleotide dissociation inhibitor (GDI) for Galphai. Recently, we used a protein delivery reagent, Chariot, and demonstrated that the GDI activity of human Ngb is tightly correlated with its neuroprotective activity. In the present study, we found that chimeric ZHHH Ngb, in which module M1 of human Ngb is replaced by that of zebrafish Ngb, protects PC12 cells against oxidative stress-induced cell death even in the absence of Chariot. Using fluorescein isothiocyanate (FITC)-labeled Ngb proteins, we demonstrated that both zebrafish and chimeric ZHHH Ngb can penetrate cell membranes in the absence of Chariot, suggesting that module M1 of zebrafish Ngb can translocate into cells. This is the first report of a native cell-membrane-penetrating globin. PMID- 18416561 TI - The folding kinetics of the SDS-induced molten globule form of reduced cytochrome c. AB - The folding of reduced cytochrome c (redcyt c) is increasingly being recognized as undergoing a mechanism that deviates from a two-state process. In previous far UV TRORD studies of redcyt c folding, a rapidly forming intermediate was attributed to the appearance of a molten-globule-like (MG) state [Chen, E., Goldbeck, R. A., and Kliger, D. S. (2003) J. Phys. Chem. A 107, 8149-8155]. A slow folding phase (>1 ms) was identified with the formation of native (N) secondary structure from that MG form. Here, using 0.65 mM SDS to induce the MG state in oxidized cytochrome c, folding of redcyt c was triggered with fast photoreduction and probed from early microseconds to milliseconds using far-UV TRORD spectroscopy. The kinetics of the reaction are described with a time constant of 50 +/- 16 ms, which corresponds to 1 +/- 0.6 ms upon extrapolation of the data to zero SDS concentration. The latter folding time is about 5 times faster than the calculated GuHCl-free time constant of 5.5 +/- 1.4 ms for slow phase folding obtained in our previous study. This ratio of rates would be consistent with a scenario in which 20-30% MG that is suggested to form in the fast phase of redcyt c folding in GuHCl is an obligatory intermediate. The native state forms from this obligatory intermediate with an observed rate, k(f) = fk(G- >N) where f is the fractional population of MG and k(G-->N) is the microscopic rate for MG --> N. Calculation and comparison of the m(#)/m values show agreement within the uncertainties between the SDS ( approximately 0.5) and GuHCl ( approximately 0.3) based redcyt c folding experiments, suggesting that the two experiments report on comparable intermediates. The m values were obtained from far-UV CD SDS titration experiments, from which calculated thermodynamic parameters allowed estimation of the reduction potential for the MG state to be approximately 155 mV (-15 kJ/mol) vs NHE which, like the reduction potential for the native state, is more favorable than that for the unfolded protein. PMID- 18416562 TI - Structural basis for the unusual specificity of Escherichia coli aminopeptidase N. AB - Aminopeptidase N from Escherichia coli is a M1 class aminopeptidase with the active-site region related to that of thermolysin. The enzyme has unusual specificity, cleaving adjacent to the large, nonpolar amino acids Phe and Tyr but also cleaving next to the polar residues Lys and Arg. To try to understand the structural basis for this pattern of hydrolysis, the structure of the enzyme was determined in complex with the amino acids L-arginine, L-lysine, L-phenylalanine, L-tryptophan, and L-tyrosine. These amino acids all bind with their backbone atoms close to the active-site zinc ion and their side chain occupying the S1 subsite. This subsite is in the form of a cylinder, about 10 A in cross-section and 12 A in length. The bottom of the cylinder includes the zinc ion and a number of polar side chains that make multiple hydrogen-bonding and other interactions with the alpha-amino group and the alpha-carboxylate of the bound amino acid. The walls of the S1 cylinder are hydrophobic and accommodate the nonpolar or largely nonpolar side chains of Phe and Tyr. The top of the cylinder is polar in character and includes bound water molecules. The epsilon-amino group of the bound lysine side chain and the guanidinium group of arginine both make multiple hydrogen bonds to this part of the S1 site. At the same time, the hydrocarbon part of the lysine and arginine side chains is accommodated within the nonpolar walls of the S1 cylinder. This combination of hydrophobic and hydrophilic binding surfaces explains the ability of ePepN to cleave Lys, Arg, Phe, and Tyr. Another favored substrate has Ala at the P1 position. The short, nonpolar side chain of this residue can clearly be bound within the hydrophobic part of the S1 cylinder, but the reason for its facile hydrolysis remains uncertain. PMID- 18416563 TI - Multiplexed proteomics mapping of yeast RNA polymerase II and III allows near complete sequence coverage and reveals several novel phosphorylation sites. AB - The multisubunit RNA polymerases (Pols) II and III synthesize mainly eukaryotic mRNAs and tRNAs, respectively. Pol II and Pol III are protein complexes consisting of 12 and 17 subunits. Here we analyzed both yeast Pol II and Pol III by multiplexed mass spectrometric analysis using various proteases and both collision induced and electron transfer dissociation. The cumulative data obtained from using the various proteases (trypsin, chymotrypsin, Glu-C and Lys C) and the two peptide fragmentation approaches allowed us to map nearly the complete sequences of all constituents of both Pol II and III. Notably, chymotrypsin behaved equally well as and in certain circumstances better than trypsin in the context of protein coverage. Although the available high resolution structures have exposed extensive mechanistic insights into transcription, the role of post-translational modification in these processes has been addressed to a lesser extent. In our analysis of Pol II and III we detected 19 phosphorylation sites, of which 12 have not been previously reported. Identified phosphosites were mapped on the Pol II structure which provided indications that they might play a role in regulating the conformation of the clamp region and, as a consequence, interaction of Pol II with nucleic acids. The described multiplexed proteomics approach is generic and reveals that it is possible to map a protein complex to near completion while applying less than 5 mug (approximately 10 pmol) of total starting material. PMID- 18416564 TI - Reversible addition-fragmentation chain transfer polymerization in DNA biosensing. AB - Reversible addition-fragmentation chain transfer polymerization is employed here to allow detector-free visualization of specific DNA sequences for which dynamic polymer growth is used in signal amplification. In particular, surface-initiated polymer growth was regulated by the immobilization of chain transfer agents on the Au surface where DNA hybridization occurred. A linear polymer growth was observed as a function of the reaction time, characteristic of "living" polymer reactions. Significant improvement in assay sensitivity was realized in comparison to the previously reported polymerization-based sensing method by enhancing polymer growth rate and reducing background noises caused by nonspecific adsorption. Direct visualization of fewer than 2,000 copies of a short oligonucleotide sequence was demonstrated in a detector-free fashion. PMID- 18416565 TI - Ceramic nanoparticle/monodisperse latex coatings. AB - Ceramic nanoparticle/monodisperse latex coatings with a nanoparticle-rich surface and a latex-rich body were created by depositing aqueous dispersions of monodisperse latex, approximately 550 nm in diameter, and nanosized ceramic particles onto substrates and drying. On the top surface of the dried coating, the latex particles are closely packed with nanoparticles uniformly occupying the interstitial spaces, and along the cross section, nanoparticles fill the spaces between the latex particles in the near surface region; a compacted latex structure, nearly devoid of nanoparticles, lies beneath. Cryogenic scanning electron microscopy images of partially dried coatings at successive drying stages reveal two important steps in forming this structure: top-down consolidation of latex particles and accumulation of nanoparticles in interstitial spaces among latex particles near the surface. A systematic study of the effect of processing conditions, including nanoparticle concentration, nanoparticle size, latex glass transition temperature, and drying conditions, on the final microstructure was carried out. The unique microstructure described above forms when the monodisperse latex is large enough to create pore channels for the transport of nanosized particles and the drying conditions favor "top down" as opposed to "edge-in" drying. PMID- 18416566 TI - Post-translational modifications of Desulfovibrio vulgaris Hildenborough sulfate reduction pathway proteins. AB - Recent developments in shotgun proteomics have enabled high-throughput studies of a variety of microorganisms at a proteome level and provide experimental validation for predicted open reading frames in the corresponding genome. More importantly, advances in mass spectrometric data analysis now allow mining of large proteomics data sets for the presence of post-translational modifications (PTMs). Although PTMs are a critical aspect of cellular activity, such information eludes cell-wide studies conducted at the transcript level. Here, we analyze several mass spectrometric data sets acquired using two-dimensional liquid chromatography tandem mass spectrometry, 2D-LC/MS/MS, for the sulfate reducing bacterium, Desulfovibrio vulgaris Hildenborough. Our searches of the raw spectra led us to discover several post-translationally modified peptides in D. vulgaris. Of these, several peptides containing a lysine with a +42 Da modification were found reproducibly across all data sets. Both acetylation and trimethylation have the same nominal +42 Da mass, and are therefore candidates for this modification. Several spectra were identified having markers for trimethylation, while one is consistent with an acetylation. Surprisingly, these modified peptides predominantly mapped to proteins involved in sulfate respiration. Other highly expressed proteins in D. vulgaris, such as enzymes involved in electron transport and other central metabolic processes, did not contain this modification. Decoy database searches were used to control for random spectrum/sequence matches. Additional validation for these modifications was provided by alternate workflows, for example, two-dimensional gel electrophoresis followed by mass spectrometry analysis of the dissimilatory sulfite reductase gamma-subunit (DsrC) protein. MS data for DsrC in this alternate workflow also contained the +42 Da modification at the same loci. Furthermore, the DsrC homologue in another sulfate reducing bacterium, Desulfovibrio desulfuricans G20, also showed similar +42 Da modifications in the same pathway. Here, we discuss our methods and implications of potential trimethylation in the D. vulgaris sulfate reduction pathway. PMID- 18416568 TI - Control of protein interfacial affinity by nonionic cosolvents. AB - In a biological cell, proteins perform their functions in a highly complex environment comprising crowding and confinement effects as well as interactions with interfaces, cosolvents, and other biomolecules. Cosolvents can stabilize or destabilize the native folded structure of proteins in solution. In this study, we show that nonionic cosolvents also affect the interfacial affinity of proteins. We use bovine ribonuclease A and a planar silica-water interface as model system and apply neutron and optical reflectometry to analyze this system. The degree of protein adsorption and the density profile of adsorbed protein molecules were determined in the absence and the presence of cosolvents. It has been found that both the protein stabilizing glycerol and the protein destabilizing urea cause a distinct reduction in protein interfacial affinity, which may represent a rather unexpected result. However, it is suggested that different mechanisms are underlying the similar effects of glycerol and urea. PMID- 18416569 TI - Stabilizing helical polyalanine peptides with negative polarity or charge: capping with cysteine. AB - Alanine-based peptides are widely known for their propensity to form helices, whether in the gas phase or in aqueous solution. Interactions of substituent groups or peptides with the helical macrodipole may either encourage or discourage the formation or stability of a helix, depending upon the placement of these groups. We report the first study of the inductive stabilization of a number of peptides through electronegative or anionic N-terminal residue capping. Using Charmm27/CMAP equilibrium and replica-exchange (REX) molecular dynamics (MD) simulations with Generalized Born implicit solvation methods, we find that the N-terminal cysteine capping of alanine peptides strongly enhances the helicity, even allowing the helical moiety to remain at temperatures beyond the denaturing temperature. Though the overall number of hydrogen bonds is enhanced, this stabilization seems to occur indirectly through interaction with the helical macrodipole rather than as a direct result of hydrogen bonding involving the cysteine, though the nature of the hydrogen bonding changes. PMID- 18416567 TI - Global changes in and characterization of specific sites of phosphorylation in mouse and human histone H1 Isoforms upon CDK inhibitor treatment using mass spectrometry. AB - Global changes in the phosphorylation state of human H1 isoforms isolated from UL3 cells have been investigated using mass spectrometry. Relative changes in H1 phosphorylation between untreated cells and cells treated with dexamethasone or various CDK inhibitors were determined. The specific cyclin-dependent kinase consensus sites of phosphorylation on the histone H1 isoforms that show changes in phosphorylation were also investigated. Three sites of phosphorylation on histone H1.4 isoforms have been identified. PMID- 18416570 TI - Isothermal crystallization kinetics study on aqueous solution of poly(vinyl methyl ether) by FTIR and optical microscopy method. AB - A study on the isothermal crystallization kinetics of aqueous solution of poly (vinyl methyl ether) (PVME) was carried out by Fourier transform infrared (FTIR) and optical microscopy respectively. IR spectra of PVME solution were measured as a function of time under the isothermal crystallization conditions. With the process of crystallization, the phase of solution changes from transparent state to opaque one within around 1-2 min for 40 or 45 wt % PVME sample, the C-H symmetric stretching bands (nus(CH3)) shift to lower wave number 2823 cm(-1). The red shift of nus(CH3) absorption band was not observed in the transparent noncrystallization area. Using the temperature jump method, we determined the growth rate of ice crystal between the glass transition temperature Tg and the melting temperature Tm. At the different crystallization temperatures Tc, the different morphologies and dimension of ice crystal are also observed. PMID- 18416571 TI - Translational diffusion of macromolecules and nanoparticles modeled as non overlapping bead arrays in an effective medium. AB - There are three objectives to the present work. First, starting from a boundary element (BE) formulation of low Reynolds number hydrodynamics, model the translational diffusion of macromolecules modeled as an array of non-overlapping beads, and show how this approach is equivalent to previous formulations of "bead hydrodynamics" and under what conditions. Second, show how this approach can be improved upon by accounting for the variation in forces over the surfaces of individual beads and also extending the approach to a gel modeled as an effective medium, EM. Third, develop a "combined obstruction and hydrodynamic effect" model of the translational diffusion of irregularly shaped macromolecules in a gel. In one of the cases studied, the BE approach is shown to be equivalent to previous "bead model" formulations in which intersubunit hydrodynamic interaction is modeled using the Rotne-Prager tensor. A bead model that accounts for the variation in hydrodynamic stress forces over the individual bead surfaces is shown to be in best agreement with exact results for simple bead arrays made up of 2-4 subunits. The translational diffusion of rods, modeled as strings of from 2 to 100 touching beads in dilute gels is examined. Interpolation formulas valid over a range of gel concentrations and rod lengths are derived for the parallel and perpendicular components of the diffusion tensor as well as the orientationally averaged diffusion tensor. The EM model accounts for the long range hydrodynamic interaction exerted by the gel support matrix on the diffusing particle of interest but does not account for the reduction in diffusion caused by the direct obstruction of the gel, or steric effect. Both effects are accounted for by writing the translational diffusion in a gel as the product of two terms representing long-range hydrodynamic interaction and steric effects. Finally, the diffusion of a 564 base pair DNA in a 2% agarose gel is examined and model results are compared to experiment (Pluen, A.; Netti, P. A.; Jain, R. K.; Berk, D. A. Biophys. J. 1999, 77, 542-552). For reasonable choices of model parameters, fair agreement between theory and experiment is achieved. PMID- 18416572 TI - Gaining an insight into the photoreactivity of a drug in a protein environment: a case study on nalidixic acid and serum albumin. AB - The binding of nalidixic acid (NA) with human and bovine serum albumin (HSA and BSA) in buffer solution at pH 7.4 was investigated using circular dichroism (CD), UV absorption and fluorescence spectroscopy. Global analysis of multiwavelength spectroscopic data afforded the equilibrium constants of the most stable noncovalent drug/protein adducts of 1:1 and 2:1 stoichiometry and their individual CD, UV absorption, and fluorescence spectra. The primary binding site of the drug was located in subdomain IIIA (Sudlow Site II), whereas the secondary one was assigned to subdomain IIA. Conformational and CD calculations afforded the binding geometries. In the complexes, the fluorescence of the protein was strongly quenched by energy transfer and that of the drug was suppressed by electron transfer. Laser flash photolysis at 355 nm evidenced the formation of a radical pair consisting of a tyroxyl radical (lambdamax = 410 nm) and a reduced nalidixate anion radical NA(2-)* (lambdamax = 640 nm) with quantum yield of 0.4 0.5. Strong evidence was obtained that the process that involves Tyr411 in HSA (Tyr409 in BSA). A further transient with lambdamax approximately 780 nm observed in HSA was attributed to oxidation of the -(S200-S246)- bridge upon electron transfer to NA(-)*. Decay of the confined radical pairs occurred with rates approximately 10(7) s(-1). Formation of covalent drug-protein adducts in mixtures irradiated at lambdairr> 324 nm was proved using HPLC with fluorescence detection. PMID- 18416573 TI - Rapid photodynamics of vitamin B6 coenzyme pyridoxal 5'-phosphate and its Schiff bases in solution. AB - The active form of vitamin B6, pyridoxal 5'-phosphate (PLP), is an important cofactor for numerous enzymes in amine and amino acid metabolism. Presented here is the first femtosecond transient absorption study of free PLP and two Schiff bases, PLP-valine and PLP-alpha-aminoisobutyric acid (AIB), in solution. Photoexcitation of free PLP leads to efficient triplet formation with an internal conversion rate that increases with increasing pH. The measured excited-state kinetics of the PLP-valine Schiff base exhibits a dramatic deuterium dependence as a result of excited-state proton transfer (ESPT) of the Calpha hydrogen in the amino acid substrate. This is consistent with formation of the key reaction carbanionic intermediate (quinonoid), which is resonance stabilized by the electron-deficient, conjugated pi system of the Schiff base/pyridine ring. The transient absorption signals of the PLP-Schiff base with alpha-methylalanine (2 aminoisobutyric acid), which does not have a Calpha proton, lack an observable deuterium effect, verifying ESPT formation of the quinonoid intermediate. In contrast to previous studies, no dependence on the excitation wavelength of the femtosecond kinetics is observed with PLP or PLP-valine, which suggests that a rapid (<250 fs) tautomerization occurs between the enolimine (absorbing at 330 nm) and ketoenamine (absorbing at 410 nm) tautomers in solution. PMID- 18416574 TI - Modular synthesis of 5-substituted thiophen-2-yl C-2'-deoxyribonucleosides. AB - A new modular methodology of preparation of 5-substituted thiophene-2-yl C nucleosides was developed. A Friedel-Crafts-type of C-glycosidation of 2 bromothiophene with toluoyl-protected methylglycoside 2 gave the desired protected 1beta-(5-bromothiophen-2-yl)-1,2-dideoxyribofuranose 4a in 60%. The key intermediate 4a was then subjected to a series of palladium-catalyzed cross coupling reactions. The cross-coupling reactions with alkyl organometallics gave beta-(5-alkylthiophen-2-yl)-2-deoxyribonucleosides 4 and 7 in moderate yields accompanied by side-products of reduction. On the other hand, cross-couplings with arylstannanes proceeded smoothly to give a series of beta-(5-arylthiophen-2 yl)-2-deoxyribonucleosides 4 in good yields. Deprotection of toluoylated nucleosides by NaOMe in MeOH and silylated nucleosides by Et 3N.3HF gave a series of free C-nucleosides 6. Alternatively, other types of 5-arylthiophene C nucleosides 6 were prepared in one step by the aqueous-phase cross-coupling reactions of unprotected 1beta-(5-bromothiophen-2-yl)-1,2-dideoxyribofuranose with boronic acids. Title 5-arylthiophene C-nucleosides 6 exhibit interesting fluorescent properties with emission maxima varying from 339 to 396 nm depending on the aryl group attached. PMID- 18416576 TI - Iminodiaziridines by regio- and stereoselective cyclization of diastereomeric singlet triazatrimethylenemethane diradicals generated through photolysis of 5 imino-4,5-dihydro-1H-tetrazoles. AB - 1,4-Dialkyl-5-(N-alkylimino)-4,5-dihydro-1H-tetrazoles were prepared in high yields by deprotonation with sodium hydride of 1,4-dialkyl-5-(N alkylamino)tetrazolium salts that were adorned with two or three different alkyl groups, including methyl, trideuteriomethyl, and tert-butyl groups. Direct irradiation (lambda > 255 nm) at -60 degrees C yielded molecular nitrogen and mixtures of 1,2-dialkyl-3-(N-alkylimino)diaziridines (83-87%) along with carbodiimides (13-17%) arising by 1,3-dipolar cycloreversion. The missing 1,3 dipoles, alkyl azides, did not survive photolysis. Each member of a pair of isotopomers and of a pair of isomers, and an iminodihydrotetrazole, whose three nitrogens were tagged, yielded a characteristic mixture of three isomeric iminodiaziridines that allowed the mode of formation to be deduced. The results are interpreted in terms of photodenitrogenation of the iminodihydrotetrazoles to furnish diastereomeric singlet triazatrimethylenemethane diradicals that retain the inherited configurations before ring closure to iminodiaziridines, presumably in two steps via mono-orthogonal diradicals. PMID- 18416575 TI - Scalable preparation of both enantiomers of 2-(1-hydroxy-2-oxocyclohexyl)acetic acid. AB - The efficient, scalable preparation of both enantiomers of 2-(1-hydroxy-2 oxocyclohexyl)acetic acid in enantiomerically pure form is reported using environmentally benign conditions in 30% overall yield (6 steps) for the (S) isomer, in 27% (7 steps) for the (R)-isomer, from cyclohexanone. PMID- 18416577 TI - Design of a "new motif" with beta-amino acids and alpha-aminoxy acids: synthesis of hybrid peptides with 12/10-helix. AB - Hybrid peptides are prepared from a C-linked carbo-beta-amino acid ester (R-beta Caa) and an alpha-aminoxy acid (R-Ama) derived from S-lactic acid. Extensive NMR (in CDCl 3 solution), CD, and MD studies on the tetra- and hexapeptides led to identification of robust 12/10-mixed helices. The dipeptide repeat having an R beta-Caa and an R-Ama thus provides a "new motif" to realize a 12/10-mixed helix, for the first time, in oligomers containing R-Ama. To understand the impact of side chains in the mixed helix formation, R-beta-Caa/Ama (with no substitution in Ama) and S-beta-hAla/R-Ama oligomers were investigated. NMR studies revealed the existence of 12/10-helices in these hybrid peptides, and the side chains of monomers were found to have a profound influence on their stabilities. These observations imply that the propensity of beta-amino acid to prefer a mixed 12/10 helix governs the structural behavior in these peptides. The structural consequences of the lone-pair repulsion between nitrogen and oxygen atoms result in a new and interesting structural motif which behaves like "pseudo" beta (3),beta(2)-peptides in generating 12/10-mixed helices. PMID- 18416578 TI - A cross-metathesis route to the 5-F(2)-isoprostanes. AB - A library of eight 5-F(2)-isoprostanes was prepared through a ring-opening metathesis/cross-metathesis protocol between functionalized bicyclo[3.2.0]heptenes, ethylene, and alpha,beta-unsaturated ketones. This sequence provided racemic enones in a regio- and stereoselective fashion that could be converted to enantiomerically enriched allylic alcohols through a catalyst-controlled asymmetric reduction. Completion of the sidechains, followed by global deprotection, resulted in a stereodivergent route to eight enantiomerically enriched 5-F(2)-isoprostanes. Overall, the synthesis of this library of known and anticipated lipid oxidation metabolites was achieved in 10 steps from commercially available 4-hydroxy-2-cyclopentenone. PMID- 18416579 TI - Concise, stereoselective route to the four diastereoisomers of 4-methylproline. AB - The full stereochemical characterization of 4-methylproline, a rare amino acid found in a number of peptidic secondary metabolites, has often been hindered by long reaction sequences or low stereoselectivity in the synthesis leading to reference samples. The preparation of the four diastereoisomers of 4 methylproline by a concise and stereoselective route is presented and features a six-step route with late-stage stereodivergence, good stereoselectivity for both cis- and trans-series (75% and 88% de, respectively), and good overall yields (cumulative yields of 30-40%). Additional data on the Marfey's derivatives of the stereoisomers are also presented. PMID- 18416580 TI - Nanogap detector inside nanofluidic channel for fast real-time label-free DNA analysis. AB - We report fabrication and characterization of a novel real-time, label-free DNA detector, that uses a long nanofluidic channel to stretch a DNA strand and a nanogap detector (with a gap as small as 9 nm) inside the channel to measure the electrical conduction perpendicular to the DNA backbone as it moves through the gap. We have observed electrical signals caused by 1.1 kilobase-pair (kbp) double stranded (ds)-DNA passing through the gap in the nanogap detectors with a gap equal to or less than 13 nm. PMID- 18416581 TI - ZnO nanostructures as efficient antireflection layers in solar cells. AB - An efficient antireflection coating (ARC) can enhance solar cell performance through increased light coupling. Here, we investigate solution-grown ZnO nanostructures as ARCs for Si solar cells and compare them to conventional single layer ARCs. We find that nanoscale morphology, controlled through synthetic chemistry, has a great effect on the macroscopic ARC performance. Compared with a silicon nitride (SiN) single layer ARC, ZnO nanorod arrays display a broadband reflection suppression from 400 to 1200 nm. For a tapered nanorod array with average tip diameter of 10 nm, we achieve a weighted global reflectance of 6.6%, which is superior to an optimized SiN single layer ARC. Calculations using rigorous coupled wave analysis suggest that the tapered nanorod arrays behave like modified single layer ARCs, where the tapering leads to impedance matching between Si and air through a gradual reduction of the effective refractive index away from the surface, resulting in low reflection particularly at longer wavelengths and eliminating interference fringes through roughening of the air ZnO interface. According to the calculations, we may further improve ARC performance by tailoring the thickness of the bottom fused ZnO layer and through better control of tip tapering. PMID- 18416582 TI - Therapy for Helicobacter pylori infection can be improved: sequential therapy and beyond. AB - As with other bacterial infections, successful treatment of Helicobacter pylori infections depends on the use of antibacterial agents to which the organism is susceptible. In this article, we use the proposed report card grading scheme (i.e. grade A, B, C, D, F) for the outcome of clinical trials, where intention-to treat cure rates >95% = A, 90-95% = B, 85-89% = C, 81-84% = D and <81% = F. The goal of therapy is to consistently cure >95% of patients (e.g. provide grade A results). Like tuberculosis, H. pylori infections are difficult to cure and successful treatment generally requires the administration of several antibacterial agents simultaneously. Duration of therapy is also important and depends upon whether resistance is present; 14 days is often best. With few exceptions, worldwide increasing macrolide resistance now undermines the effectiveness of the legacy triple therapy (e.g. a proton pump inhibitor [PPI], clarithromycin and amoxicillin) and, in most areas, cure rates have declined to unacceptable levels (e.g. grade F). The development of sequential therapy was one response to this problem. Sequential therapy has repeatedly been shown in head-to head studies to be superior to legacy triple therapy. Sequential therapy, as originally described, is the sequential administration of a dual therapy (a PPI plus amoxicillin) followed by a Bazzoli-type triple therapy (a PPI plus clarithromycin and tinidazole) and has been shown to be especially useful where there is clarithromycin resistance. However, the cure rates of the original sequential treatment are grade B and can probably be further improved by changes in dose, duration or administration, such as by continuing the amoxicillin into the triple therapy arm. The sequential approach may also be more complicated than necessary, based on the fact that the same four drugs have also been given concomitantly (at least nine publications with >700 patients) as a quadruple therapy with excellent success. This article discusses the approach to therapy in the modern era where antimicrobial resistance is an increasing problem and legacy triple therapy is no longer an acceptable initial choice. Methods to achieve acceptable eradication rates (e.g. grade A or B results) are discussed and, specifically, sequential therapy is considered both conceptually and practically. Suggestions are provided regarding how sequential therapy might be improved to become a grade A therapy as well as how to identify situations where it can be expected to yield unacceptable results. New uses for current drugs are discussed and suggestions for subsequent randomized comparisons to overcome phenotypic and genotypic resistance are given. We propose a change in focus from comparative studies (designed to prove that a new therapy is superior to a known inferior therapy) to demanding that efficacious therapies meet or exceed a pre-specified level of success (i.e. grade A or B result). To do so, coupled with less concern about the effect of recommendations on the pharmaceutical industry, should provide clinicians with much higher quality information, and improve the quality of medical care and recommendations regarding treatment. Ultimately, there is little or no justification for comparative testing that includes an arm with known unacceptably low results. H. pylori gastritis is an infectious disease and should be approached and treated as such. PMID- 18416583 TI - Bevacizumab in non-small cell lung cancer. AB - Lung cancer continues to be the leading cause of cancer death in Western countries. The median survival time for advanced non-small cell lung cancer (NSCLC) remains poor and chemotherapy is the treatment of choice for most patients with metastatic NSCLC. Platinum-based chemotherapy has long been the standard of care for advanced NSCLC. The formation of new blood vessels (angiogenesis) is needed for the growth and invasiveness of primary tumours, and plays an important role in metastatic growth. Vascular endothelial growth factor (VEGF) has emerged as a key potential target for the pharmacological inhibition of tumour angiogenesis. This review discusses current data and the future potential of bevacizumab, a recombinant humanized monoclonal antibody that binds VEGF, in the treatment of NSCLC. Results from a phase II study showed that the addition of bevacizumab to the first-line chemotherapy with paclitaxel and carboplatin (CP) may increase the overall survival (OS) and the time to progression in advanced NSCLC. Based on these promising results, a randomized phase III trial compared the combination of bevacizumab with CP versus CP alone in the treatment of advanced non-squamous NSCLC. The combination of CP plus bevacizumab led to a statistically significant increase in median OS and progression-free survival (PFS) compared with CP alone, with a response rate (RR) in the CP arm of 15% compared with 35% in the bevacizumab plus CP arm (p < 0.001). More recently, the randomized AVAIL (Avastin in Lung Cancer) study, which evaluated cisplatin with gemcitabine plus bevacizumab in two different dosages versus chemotherapy alone in 1043 patients with recurrent or advanced non squamous NSCLC, reported a significant increase of PFS, RR and duration of response for both of the bevacizumab-containing arms. Bevacizumab has also been investigated in combination with erlitonib as second-line treatment in two small early phase trials, with interesting results. Bevacizumab was generally well tolerated in clinical trials; the main treatment-associated adverse events were neutropenia and haemorrhage, especially in the lung, but also at other sites. Several trials that incorporate bevacizumab in combination with new active drugs in NSCLC are ongoing and should further help to define the place of bevacizumab in the therapy of NSCLC. PMID- 18416584 TI - Novel therapies for anti-neutrophil cytoplasmic antibody-associated vasculitis. AB - The rapid expansion in the therapeutic modalities available for the treatment of anti-neutrophil cytoplasmic antibody-associated vasculitides (AAV), with clear limitations in existing strategies, prompted us to undertake a review of novel therapies reported in MEDLINE and EMBASE. Tumour necrosis factor (TNF)-alpha antagonism with infliximab is described favourably in retrospective series and open-label trials. However, evidence from the WGET (Wegener's Granulomatosis Etanercept Trial) does not support the clinical use of etanercept, and a significantly higher malignancy rate following TNFalpha inhibition questions the role of this approach. Uncontrolled evidence alone supports remission induction with rituximab-mediated B-lymphocyte depletion and may be less effective in predominantly granulomatous AAV. Remission following T-lymphocyte depletion can be achieved with alemtuzumab and antithymocyte globulin, but it is not yet clear what the clinical role will be for these agents in AAV. In addition, these agents are associated with prolonged lymphopenia and pulmonary complications, respectively. Stem cell transplantation to support immune reconstitution following the use of such agents has been trialled in AAV, but studies included very few patients. Purine and pyrimidine antimetabolites mycophenolate mofetil and leflunomide are likely to play an important role in the treatment of AAV, but results supporting remission maintenance and induction in the former are limited to uncontrolled trials, such that their use remains experimental at this time. Similarly, 15-deoxyspergualin may provide an alternative to cyclophosphamide but awaits randomized controlled trial evidence. The MEPEX (MEthylprednisolone versus Plasma EXchange) trial supports plasma exchange in renal disease but this may be limited by pulmonary complications. Randomized controlled evidence also exists for intravenous immunoglobulin, although improvement may not be sustained. Antimicrobial therapy may be of use in Wegener's granulomatosis patients with predominantly upper respiratory tract involvement. Safety concerns, notably of infection and malignancy, were common and need to be explored in subsequent trials. In addition, concomitant immunosuppressants and non-standardized definitions were major limitations, and future studies of these and newer agents must follow agreed standards of study design and reporting to facilitate clearer interpretation of the circumstances (e.g. disease stage, severity or organ involvement) under which these agents perform optimally. Consequently, use is still limited to centres experienced in such agents and mostly in the context of clinical trials. PMID- 18416586 TI - Early and sustained virological response in non-responders with chronic hepatitis C: a randomized open-label study of pegylated interferon-alpha-2a versus pegylated interferon-alpha-2b. AB - OBJECTIVES: The purpose of this randomized open-label study was to assess the efficacy of treatment with pegylated interferon-alpha-2a versus pegylated interferon-alpha-2b, both plus ribavirin, in inducing early and sustained virological response (EVR and SVR) in chronic hepatitis C non-responders. PATIENTS AND METHODS: A total of 108 patients with chronic hepatitis C who were non-responders to previous combined therapy (standard interferon-alpha plus ribavirin for > or = 3 months) were enrolled and equally randomized into two groups in this intention-to-treat analysis. The patients exhibited similar baseline features. One group received subcutaneous pegylated interferon-alpha-2a 180 microg once weekly, while the other was treated with subcutaneous pegylated interferon-alpha-2b 1.5 microg/kg once weekly. Ribavirin 15 mg/kg/day was included in both protocols. Treatment duration for EVR was 12 weeks. Patients who demonstrated non-detectable hepatitis C virus (HCV) RNA or a > or = 2 log(10) reduction in viral load at week 12 continued therapy up to 48 weeks, with assessments every 3 months during a follow-up of 24 weeks. RESULTS: All patients in both groups completed the EVR study, then seven patients receiving pegylated interferon-alpha-2a and seven patients receiving pegylated interferon-alpha2b discontinued treatment as a result of severe adverse effects. After 12 weeks of treatment, viral load reduction was >2 log(10) with both pegylated interferon alpha-2a (-2.53) and pegylated interferon-alpha-2b (-2.48) with no significant difference. At the end of week 48, HCV RNA was undetectable in 14 of 54 patients (25.9%) receiving pegylated interferon-alpha-2a and in 15 of 54 patients (27.7%) receiving pegylated interferon-alpha-2b. When terminating follow-up, an SVR was observed in 11 of 54 patients (20.4%) who received pegylated interferon-alpha-2a and 10 of 54 patients (18.4%) receiving pegylated interferon-alpha-2b. The incidence and severity of adverse events was similar in both groups. CONCLUSIONS: Our results seem to show that in chronic hepatitis C patients who are non responsive to previous therapy, EVR to the two pegylated interferons did not significantly differ with a similar therapeutic efficacy defined as SVR. PMID- 18416585 TI - Pharmaceutical management of ovarian cancer : current status. AB - Over recent decades, truly impressive progress has been made in the outcome associated with the pharmacological antineoplastic management of women with advanced ovarian cancer. Following initial surgery, the large majority of patients with this malignancy will receive a chemotherapy regimen that includes a platinum drug (carboplatin or cisplatin) and a taxane (paclitaxel or docetaxel). Currently, objective responses are observed in approximately 60-80% of patients treated in the front-line setting, with documented improvements in overall survival compared with prior non-platinum and taxane programmes. Unfortunately, despite the high response rate to initial chemotherapy, the majority of women with advanced disease will experience recurrence of the malignant process and be candidates for a variety of possible second-line therapeutic options. It is well recognized that ovarian cancer patients who are documented to experience an initial response to platinum-based chemotherapy but where the disease recurs approximately 6 or more months following the completion of primary therapy, may have another clinically meaningful response (both objective and subjective) to a second platinum-based strategy. However, an optimal management approach in this setting remains to be defined. Furthermore, the malignant cell populations in all ovarian cancer patients who experience an initial relapse of the disease process will eventually be resistant to the platinum agents. In this setting, multiple drugs have been shown to be biologically active. Again, an optimal strategy to be employed in the platinum-resistant setting has yet to be demonstrated through the conduct of evidence-based trials. Reasonable goals of therapy in women with recurrent or resistant ovarian cancer are to improve overall survival, reduce the severity (and delay the occurrence) of symptoms and optimize overall quality of life. PMID- 18416587 TI - Meropenem: a review of its use in the treatment of serious bacterial infections. AB - Meropenem (Merrem, Meronem) is a broad-spectrum antibacterial agent of the carbapenem family, indicated as empirical therapy prior to the identification of causative organisms, or for disease caused by single or multiple susceptible bacteria in both adults and children with a broad range of serious infections. Meropenem is approved for use in complicated intra-abdominal infection (cIAI), complicated skin and skin structure infection (cSSSI) and bacterial meningitis (in paediatric patients aged > or = 3 months) in the US, and in most other countries for nosocomial pneumonia, cIAI, septicaemia, febrile neutropenia, cSSSI, bacterial meningitis, complicated urinary tract infection (UTI), obstetric and gynaecological infections, in cystic fibrosis patients with pulmonary exacerbations, and for the treatment of severe community-acquired pneumonia (CAP). Meropenem has a broad spectrum of in vitro activity against Gram-positive and Gram-negative pathogens, including extended-spectrum beta-lactamase (ESBL)- and AmpC-producing Enterobacteriaceae. It has similar efficacy to comparator antibacterial agents, including: imipenem/cilastatin in cIAI, cSSSI, febrile neutropenia, complicated UTI, obstetric or gynaecological infections and severe CAP; clindamycin plus tobramycin or gentamicin in cIAI or obstetric/gynaecological infections; cefotaxime plus metronidazole in cIAI; cefepime and ceftazidime plus amikacin in septicaemia or febrile neutropenia; and ceftazidime, clarithromycin plus ceftriaxone or amikacin in severe CAP. Meropenem has also shown similar efficacy to cefotaxime in paediatric and adult patients with bacterial meningitis, and to ceftazidime when both agents were administered with or without tobramycin in patients with cystic fibrosis experiencing acute pulmonary exacerbations. Meropenem showed greater efficacy than ceftazidime or piperacillin/tazobactam in febrile neutropenia, and greater efficacy than ceftazidime plus amikacin or tobramycin in patients with nosocomial pneumonia. Meropenem is well tolerated and has the advantage of being suitable for administration as an intravenous bolus or infusion. Its low propensity for inducing seizures means that it is suitable for treating bacterial meningitis and is the only carbapenem approved in this indication. Thus, meropenem continues to be an important option for the empirical treatment of serious bacterial infections in hospitalized patients. PMID- 18416588 TI - Sucrose-formulated octocog alfa: a review of its use in patients with haemophilia A. AB - Sucrose-formulated octocog alfa (Kogenate Bayer, Kogenate FS, Helixate FS, Helixate nexgen) is a full-length recombinant human coagulation factor VIII (FVIII) product that is purified and formulated without the addition of human serum albumin and is stabilized with sucrose. The purification process of this formulation includes a solvent/detergent viral inactivation step. Sucrose formulated octocog alfa is approved in the EU and US for the treatment of bleeding in patients with haemophilia A (congenital FVIII deficiency). Additionally, it is approved in the EU for the prophylaxis of bleeding in patients with haemophilia A and as a continuous infusion treatment in patients undergoing major surgery. Sucrose-formulated octocog alfa is effective and well tolerated as a FVIII replacement therapy in patients with haemophilia A, including those with severe disease undergoing major surgery. The therapeutic profile of this sucrose-formulated product cannot be compared with that of other octocog alfa or moroctocog alfa products because of a lack of head-to-head comparative studies. Pathogen transmission has not been reported with use of sucrose-formulated octocog alfa. Available data indicate that sucrose-formulated octocog alfa is an appropriate alternative to other recombinant FVIII products for the treatment and prophylaxis of bleeding episodes in adults and children with haemophilia A. PMID- 18416589 TI - Retapamulin: a review of its use in the management of impetigo and other uncomplicated superficial skin infections. AB - Topical retapamulin (Altabax, Altargo) is the first pleuromutilin antibacterial approved for the treatment of uncomplicated superficial skin infections caused by Staphylococcus aureus (excluding meticillin-resistant S. aureus [MRSA]) and Streptococcus pyogenes in patients aged > or = 9 months. In the EU, retapamulin is indicated for use in patients with impetigo or with infected small lacerations, abrasions or sutured wounds (without abscesses); in the US, it is indicated for use in patients with impetigo. Retapamulin has a novel site of action on bacterial ribosomes. In clinical trials in patients with impetigo, topical retapamulin 1% ointment twice daily for 5 days (the approved regimen) was superior to placebo; treatment with retapamulin was noninferior to that with topical fusidic acid. In patients with secondarily infected traumatic lesions, treatment with retapamulin was noninferior to that with oral cefalexin, although the efficacy of retapamulin was reduced in patients with MRSA infections or superficial abscesses. Retapamulin was well tolerated in both paediatric and adult patients, and the majority of adverse events were of mild to moderate severity. Thus, the introduction of topical retapamulin 1% ointment extends the treatment options available in the management of impetigo and uncomplicated secondarily infected traumatic lesions. PMID- 18416590 TI - Ciclesonide nasal spray: in allergic rhinitis. AB - Ciclesonide nasal spray delivers the corticosteroid ciclesonide as a hypotonic spray via a metered-dose manual pump. Systemic exposure to ciclesonide and its active metabolite desisobutyryl-ciclesonide is low after intranasal administration. High protein binding (approximately 99%) and rapid first-pass clearance further reduce systemic exposure to the drug. In well designed trials, intranasal ciclesonide 200 microg once daily for 2-4 weeks was more effective than placebo in terms of improving nasal symptoms in adolescents and adults with moderate to severe seasonal allergic rhinitis. Quality of life measures were statistically significantly improved in ciclesonide relative to placebo recipients during the first 2 weeks of therapy. Similarly, in adolescents and adults with moderately severe perennial allergic rhinitis, ciclesonide 200 microg once daily was more effective than placebo in terms of reducing nasal symptoms in well designed trials of 6 weeks' and 1 year's duration. Improvements relative to placebo in quality of life measures were not considered clinically relevant. Ciclesonide nasal spray was generally well tolerated in these clinical trials; most adverse events were mild to moderate in intensity. PMID- 18416591 TI - Muscle fatigue during football match-play. AB - One of the consequences of sustaining exercise for 90 minutes of football match play is that the capability of muscle to generate force declines. This impairment is reflected in the decline of work-rate towards the late part of the game. Causes of this phenomenon, which is known as fatigue, and some of its consequences are considered in this article. The stores of muscle glycogen may be considerably reduced by the end of the game, especially if there has not been a tapering of the training load. Thermoregulatory strain may also be encountered, resulting in a fall in physical performance, or there may be a reduced central drive from the nervous system. The decline in muscle strength may increase the predisposition to injury in the lower limbs. Central fatigue may also occur with implications for muscle performance. Strategies to offset fatigue include astute use of substitutions, appropriate nutritional preparation and balancing pre cooling and warm-up procedures. There is also a role for endurance training and for a pacing strategy that optimizes the expenditure of energy during match-play. PMID- 18416592 TI - Scapular positioning in athlete's shoulder : particularities, clinical measurements and implications. AB - Despite the essential role played by the scapula in shoulder function, current concepts in shoulder training and treatment regularly neglect its contribution. The 'scapular dyskinesis' is an alteration of the normal scapular kinematics as part of scapulohumeral rhythm, which has been shown to be a nonspecific response to a host of proximal and distal shoulder injuries. The dyskinesis can react in many ways with shoulder motion and function to increase the dysfunction. Thoracic kyphosis, acromio-clavicular joint disorders, subacromial or internal impingement, instability or labral pathology can alter scapular kinematics. Indeed, alteration of scapular stabilizing muscle activation, inflexibility of the muscles and capsule-ligamentous complex around the shoulder may affect the resting position and motion of the scapula. Given the interest in the scapular positioning and patterns of motion, this article aims to give a detailed overview of the literature focusing on the role of the scapula within the shoulder complex through the sports context. Such an examination of the role of the scapula requires the description of the normal pattern of scapula motion during shoulder movement; this also implies the study of possible scapular adaptations with sports practice and scapular dyskinesis concomitant to fatigue, impingement and instability. Different methods of scapular positioning evaluation are gathered from the literature in order to offer to the therapist the possibility of detecting scapular asymmetries through clinical examinations. Furthermore, current concepts of rehabilitation dealing with relieving symptoms associated with inflexibility, weakness or activation imbalance of the muscles are described. Repeating clinical assessments throughout the rehabilitation process highlights improvements and allows the therapist to actualize rationally his or her intervention. The return to the field must be accompanied by a transitory phase, which is conducive to integrating new instructions during sports gestures. On the basis of the possible scapular disturbance entailed in sports practice, a preventive approach that could be incorporated into training management is encouraged. PMID- 18416593 TI - The role of membrane fatty-acid transporters in regulating skeletal muscle substrate use during exercise. AB - While endogenous carbohydrates form the main substrate source during high intensity exercise, long-chain fatty acids (LCFA) represent the main substrate source during more prolonged low- to moderate-intensity exercise. Adipose tissue lipolysis is responsible for the supply of LCFA to the contracting muscle. Once taken up by skeletal muscle tissue, LCFA can either serve as a substrate for oxidative phosphorylation or can be directed towards esterification into triacylglycerol. Myocellular uptake of LCFA comprises a complex and incompletely understood process. Although LCFA can enter the cell via passive diffusion, more recent reports indicate that LCFA uptake is tightly regulated by plasma membrane located transport proteins (fatty acid translocase [FAT/CD36], plasmalemmal located fatty acid binding protein [FABPpm] and fatty acid transport protein [FATP]). Depending on cardiac and skeletal muscle energy demands, some of these LCFA transporters can translocate rapidly from intracellular pools to the plasma membrane to allow greater LCFA uptake. This translocation process can be induced by insulin and/or muscle contraction. However, the precise signalling pathways responsible for activating the translocation machinery remain to be elucidated. This article will provide an overview on the effects of diet, acute exercise and exercise training on the expression and/or translocation of the various LCFA transporters in skeletal muscle tissue (FAT/CD36, FABPpm, FATP). PMID- 18416596 TI - Caution is required when comparing the effectiveness of voluntary versus stimulated versus combined strength training modalities. PMID- 18416595 TI - Cardiac troponin T release after prolonged strenuous exercise. AB - Over the past 2 decades, there has been a large interest in cardiac troponin T (cTnT) elevations, which are often seen following endurance sport events. There have been many reports on this topic, although sometimes with different approaches. We reviewed the available literature on cTnT elevations after prolonged strenuous exercise and discovered profound differences in the percentage of subjects reported to have elevated cTnT concentrations. This could partly be attributed to differences in immunoassay characteristics, such as cross reactivity with skeletal troponin T, and the use of different cut-off values used in the different studies. The elevations were transient, with levels decreasing to pre-event concentrations within 24-48 hours. This might be explained by the relatively short half-life of cTnT, or water imbalance during and after the event. The release mechanism of cTnT, as well as the long-term positive or negative effects, remains unclear. Future research should therefore be aimed at clarifying the release mechanism of cTnT. Furthermore, the benefits and the possible long-term negative aspects of prolonged exercise should be evaluated. PMID- 18416594 TI - Catecholamines and the effects of exercise, training and gender. AB - Stress hormones, adrenaline (epinephrine) and noradrenaline (norepinephrine), are responsible for many adaptations both at rest and during exercise. Since their discovery, thousands of studies have focused on these two catecholamines and their importance in many adaptive processes to different stressors such as exercise, hypoglycaemia, hypoxia and heat exposure, and these studies are now well acknowledged. In fact, since adrenaline and noradrenaline are the main hormones whose concentrations increase markedly during exercise, many researchers have worked on the effect of exercise on these amines and reported 1.5 to >20 times basal concentrations depending on exercise characteristics (e.g. duration and intensity). Similarly, several studies have shown that adrenaline and noradrenaline are involved in cardiovascular and respiratory adjustments and in substrate mobilization and utilization. Thus, many studies have focused on physical training and gender effects on catecholamine response to exercise in an effort to verify if significant differences in catecholamine responses to exercise could be partly responsible for the different performances observed between trained and untrained subjects and/or men and women. In fact, previous studies conducted in men have used different types of exercise to compare trained and untrained subjects in response to exercise at the same absolute or relative intensity. Their results were conflicting for a while. As research progressed, parameters such as age, nutritional and emotional state have been found to influence catecholamine concentrations. As a result, most of the recent studies have taken into account all these parameters. Those studies also used very well trained subjects and/or more intense exercise, which is known to have a greater effect on catecholamine response so that differences between trained and untrained subjects are more likely to appear. Most findings then reported a higher adrenaline response to exercise in endurance-trained compared with untrained subjects in response to intense exercise at the same relative intensity as all-out exercise. This phenomenon is referred to as the 'sports adrenal medulla'. This higher capacity to secrete adrenaline was observed both in response to physical exercise and to other stimuli such as hypoglycaemia and hypoxia. For some authors, this phenomenon can partly explain the higher physical performance observed in trained compared with untrained subjects. More recently, these findings have also been reported in anaerobic-trained subjects in response to supramaximal exercise. In women, studies remain scarce; the results are more conflicting than in men and the physical training type (aerobic or anaerobic) effects on catecholamine response remain to be specified. Conversely, the works undertaken in animals are more unanimous and suggest that physical training can increase the capacity to secrete adrenaline via an increase of the adrenal gland volume and adrenaline content. PMID- 18416598 TI - In vivo response to methotrexate forecasts outcome of acute lymphoblastic leukemia and has a distinct gene expression profile. AB - BACKGROUND: Childhood acute lymphoblastic leukemia (ALL) is the most common cancer in children, and can now be cured in approximately 80% of patients. Nevertheless, drug resistance is the major cause of treatment failure in children with ALL. The drug methotrexate (MTX), which is widely used to treat many human cancers, is used in essentially all treatment protocols worldwide for newly diagnosed ALL. Although MTX has been extensively studied for many years, relatively little is known about mechanisms of de novo resistance in primary cancer cells, including leukemia cells. This lack of knowledge is due in part to the fact that existing in vitro methods are not sufficiently reliable to permit assessment of MTX resistance in primary ALL cells. Therefore, we measured the in vivo antileukemic effects of MTX and identified genes whose expression differed significantly in patients with a good versus poor response to MTX. METHODS AND FINDINGS: We utilized measures of decreased circulating leukemia cells of 293 newly diagnosed children after initial "up-front" in vivo MTX treatment (1 g/m(2)) to elucidate interpatient differences in the antileukemic effects of MTX. To identify genomic determinants of these effects, we performed a genome-wide assessment of gene expression in primary ALL cells from 161 of these newly diagnosed children (1-18 y). We identified 48 genes and two cDNA clones whose expression was significantly related to the reduction of circulating leukemia cells after initial in vivo treatment with MTX. This finding was validated in an independent cohort of children with ALL. Furthermore, this measure of initial MTX in vivo response and the associated gene expression pattern were predictive of long-term disease-free survival (p < 0.001, p = 0.02). CONCLUSIONS: Together, these data provide new insights into the genomic basis of MTX resistance and interpatient differences in MTX response, pointing to new strategies to overcome MTX resistance in childhood ALL. TRIAL REGISTRATIONS: Total XV, Therapy for Newly Diagnosed Patients With Acute Lymphoblastic Leukemia, http://www.ClinicalTrials.gov (NCT00137111); Total XIIIBH, Phase III Randomized Study of Antimetabolite-Based Induction plus High-Dose MTX Consolidation for Newly Diagnosed Pediatric Acute Lymphocytic Leukemia at Intermediate or High Risk of Treatment Failure (NCI-T93-0101D); Total XIIIBL, Phase III Randomized Study of Antimetabolite-Based Induction plus High-Dose MTX Consolidation for Newly Diagnosed Pediatric Acute Lymphocytic Leukemia at Lower Risk of Treatment Failure (NCI-T93-0103D). PMID- 18416599 TI - Targeted nanoparticles for imaging incipient pancreatic ductal adenocarcinoma. AB - BACKGROUND: Pancreatic ductal adenocarcinoma (PDAC) carries an extremely poor prognosis, typically presenting with metastasis at the time of diagnosis and exhibiting profound resistance to existing therapies. The development of molecular markers and imaging probes for incipient PDAC would enable earlier detection and guide the development of interventive therapies. Here we sought to identify novel molecular markers and to test their potential as targeted imaging agents. METHODS AND FINDINGS: Here, a phage display approach was used in a mouse model of PDAC to screen for peptides that specifically bind to cell surface antigens on PDAC cells. These screens yielded a motif that distinguishes PDAC cells from normal pancreatic duct cells in vitro, which, upon proteomics analysis, identified plectin-1 as a novel biomarker of PDAC. To assess their utility for in vivo imaging, the plectin-1 targeted peptides (PTP) were conjugated to magnetofluorescent nanoparticles. In conjunction with intravital confocal microscopy and MRI, these nanoparticles enabled detection of small PDAC and precursor lesions in engineered mouse models. CONCLUSIONS: Our approach exploited a well-defined model of PDAC, enabling rapid identification and validation of PTP. The developed specific imaging probe, along with the discovery of plectin-1 as a novel biomarker, may have clinical utility in the diagnosis and management of PDAC in humans. PMID- 18416600 TI - Research in complex humanitarian emergencies: the Medecins Sans Frontieres/Epicentre experience. PMID- 18416601 TI - Gene-environment interaction in yeast gene expression. AB - The effects of genetic variants on phenotypic traits often depend on environmental and physiological conditions, but such gene-environment interactions are poorly understood. Recently developed approaches that treat transcript abundances of thousands of genes as quantitative traits offer the opportunity to broadly characterize the architecture of gene-environment interactions. We examined the genetic and molecular basis of variation in gene expression between two yeast strains (BY and RM) grown in two different conditions (glucose and ethanol as carbon sources). We observed that most transcripts vary by strain and condition, with 2,996, 3,448, and 2,037 transcripts showing significant strain, condition, and strain-condition interaction effects, respectively. We expression profiled over 100 segregants derived from a cross between BY and RM in both growth conditions, and identified 1,555 linkages for 1,382 transcripts that show significant gene-environment interaction. At the locus level, local linkages, which usually correspond to polymorphisms in cis-regulatory elements, tend to be more stable across conditions, such that they are more likely to show the same effect or the same direction of effect across conditions. Distant linkages, which usually correspond to polymorphisms influencing trans-acting factors, are more condition-dependent, and often show effects in different directions in the two conditions. We characterized a locus that influences expression of many growth-related transcripts, and showed that the majority of the variation is explained by polymorphism in the gene IRA2. The RM allele of IRA2 appears to inhibit Ras/PKA signaling more strongly than the BY allele, and has undergone a change in selective pressure. Our results provide a broad overview of the genetic architecture of gene-environment interactions, as well as a detailed molecular example, and lead to key insights into how the effects of different classes of regulatory variants are modulated by the environment. These observations will guide the design of studies aimed at understanding the genetic basis of complex traits. PMID- 18416602 TI - Cooperative retraction of bundled type IV pili enables nanonewton force generation. AB - The causative agent of gonorrhea, Neisseria gonorrhoeae, bears retractable filamentous appendages called type IV pili (Tfp). Tfp are used by many pathogenic and nonpathogenic bacteria to carry out a number of vital functions, including DNA uptake, twitching motility (crawling over surfaces), and attachment to host cells. In N. gonorrhoeae, Tfp binding to epithelial cells and the mechanical forces associated with this binding stimulate signaling cascades and gene expression that enhance infection. Retraction of a single Tfp filament generates forces of 50-100 piconewtons, but nothing is known, thus far, on the retraction force ability of multiple Tfp filaments, even though each bacterium expresses multiple Tfp and multiple bacteria interact during infection. We designed a micropillar assay system to measure Tfp retraction forces. This system consists of an array of force sensors made of elastic pillars that allow quantification of retraction forces from adherent N. gonorrhoeae bacteria. Electron microscopy and fluorescence microscopy were used in combination with this novel assay to assess the structures of Tfp. We show that Tfp can form bundles, which contain up to 8 10 Tfp filaments, that act as coordinated retractable units with forces up to 10 times greater than single filament retraction forces. Furthermore, single filament retraction forces are transient, whereas bundled filaments produce retraction forces that can be sustained. Alterations of noncovalent protein protein interactions between Tfp can inhibit both bundle formation and high amplitude retraction forces. Retraction forces build over time through the recruitment and bundling of multiple Tfp that pull cooperatively to generate forces in the nanonewton range. We propose that Tfp retraction can be synchronized through bundling, that Tfp bundle retraction can generate forces in the nanonewton range in vivo, and that such high forces could affect infection. PMID- 18416603 TI - Reconstitution of DNA strand exchange mediated by Rhp51 recombinase and two mediators. AB - In the fission yeast Schizosaccharomyces pombe, genetic evidence suggests that two mediators, Rad22 (the S. pombe Rad52 homolog) and the Swi5-Sfr1 complex, participate in a common pathway of Rhp51 (the S. pombe Rad51 homolog)-mediated homologous recombination (HR) and HR repair. Here, we have demonstrated an in vitro reconstitution of the central step of DNA strand exchange during HR. Our system consists entirely of homogeneously purified proteins, including Rhp51, the two mediators, and replication protein A (RPA), which reflects genetic requirements in vivo. Using this system, we present the first robust biochemical evidence that concerted action of the two mediators directs the loading of Rhp51 onto single-stranded DNA (ssDNA) precoated with RPA. Dissection of the reaction reveals that Rad22 overcomes the inhibitory effect of RPA on Rhp51-Swi5-Sfr1 mediated strand exchange. In addition, Rad22 negates the requirement for a strict order of protein addition to the in vitro system. However, despite the presence of Rad22, Swi5-Sfr1 is still essential for strand exchange. Importantly, Rhp51, but neither Rad22 nor the Swi5-Sfr1 mediator, is the factor that displaces RPA from ssDNA. Swi5-Sfr1 stabilizes Rhp51-ssDNA filaments in an ATP-dependent manner, and this stabilization is correlated with activation of Rhp51 for the strand exchange reaction. Rad22 alone cannot activate the Rhp51 presynaptic filament. AMP-PNP, a nonhydrolyzable ATP analog, induces a similar stabilization of Rhp51, but this stabilization is independent of Swi5-Sfr1. However, hydrolysis of ATP is required for processive strand transfer, which results in the formation of a long heteroduplex. Our in vitro reconstitution system has revealed that the two mediators have indispensable, but distinct, roles for mediating Rhp51 loading onto RPA-precoated ssDNA. PMID- 18416604 TI - Polycomb repressive complex 2 (PRC2) restricts hematopoietic stem cell activity. AB - Polycomb group proteins are transcriptional repressors that play a central role in the establishment and maintenance of gene expression patterns during development. Using mice with an N-ethyl-N-nitrosourea (ENU)-induced mutation in Suppressor of Zeste 12 (Suz12), a core component of Polycomb Repressive Complex 2 (PRC2), we show here that loss of Suz12 function enhances hematopoietic stem cell (HSC) activity. In addition to these effects on a wild-type genetic background, mutations in Suz12 are sufficient to ameliorate the stem cell defect and thrombocytopenia present in mice that lack the thrombopoietin receptor (c-Mpl). To investigate the molecular targets of the PRC2 complex in the HSC compartment, we examined changes in global patterns of gene expression in cells deficient in Suz12. We identified a distinct set of genes that are regulated by Suz12 in hematopoietic cells, including eight genes that appear to be highly responsive to PRC2 function within this compartment. These data suggest that PRC2 is required to maintain a specific gene expression pattern in hematopoiesis that is indispensable to normal stem cell function. PMID- 18416605 TI - Host PrP glycosylation: a major factor determining the outcome of prion infection. AB - The expression of the prion protein (PrP) is essential for transmissible spongiform encephalopathy (TSE) or prion diseases to occur, but the underlying mechanism of infection remains unresolved. To address the hypothesis that glycosylation of host PrP is a major factor influencing TSE infection, we have inoculated gene-targeted transgenic mice that have restricted N-linked glycosylation of PrP with three TSE strains. We have uniquely demonstrated that mice expressing only unglycosylated PrP can sustain a TSE infection, despite altered cellular location of the host PrP. Moreover we have shown that brain material from mice infected with TSE that have only unglycosylated PrP(Sc) is capable of transmitting infection to wild-type mice, demonstrating that glycosylation of PrP is not essential for establishing infection within a host or for transmitting TSE infectivity to a new host. We have further dissected the requirement of each glycosylation site and have shown that different TSE strains have dramatically different requirements for each of the glycosylation sites of host PrP, and moreover, we have shown that the host PrP has a major role in determining the glycosylation state of de novo generated PrP(Sc). PMID- 18416606 TI - Bond failure with a no-mix adhesive system. AB - OBJECTIVE: To determine the bond failure pattern and time to first bond failure at an orthodontic clinic. MATERIALS AND METHODS: The study included 63 subjects meeting the study criteria. A total of 1074 brackets (Roth prescription 0.022 slot) were bonded to incisors, canines, and premolars using a no-mix adhesive (Unite, 3M Unitek). The survival and failure rates of the brackets were evaluated by tooth position in the dental arch, sagittal occlusal relationship, and gender of the patients. Overall bracket survival rates were estimated using the Kaplan Meier test. RESULTS: The total percentage of bond failure was 17.87%. The mean survival time for the sample was 235 days (SE = 32.27 days). Significantly higher failure rates were observed for posterior than anterior teeth (P < .05) and in the Class II division 2 malocclusion group than other malocclusion groups (P < .05). No difference was observed between dental arches or genders. However, in terms of survival time, the difference was marginally significant for gender (P = .051) CONCLUSION: For 1074 brackets bonded with a no-mix adhesive system in 63 patients, the mean time until first bracket failure was 235 days. The overall failure rate of brackets was 17.87%. PMID- 18416607 TI - Time course of expression of bcl-2 and bax in rabbit condylar chondrocytes following forward mandibular positioning. AB - OBJECTIVE: To clarify the expression of Bcl-2 and Bax following forward mandibular positioning (FMP) in the condylar chondrocytes of rabbits. MATERIALS AND METHODS: Sixty rabbits at 8 weeks of age were randomly allocated to the experimental group (n = 36) and control group (n = 24). Rabbits in the experimental group were induced to FMP by a functional appliance. Six rabbits from the experimental group and four from the control group were sacrificed after 3 days and 1, 2, 4, 8, and 12 weeks, respectively. All the right temporomandibular joints (TMJs) were collected and the expression of Bcl-2 and Bax was evaluated by immunohistochemical staining. RESULTS: The results showed the expression pattern of Bcl-2 and Bax during 12 weeks after FMP. The expression of Bcl-2 reached the highest level at 1 week, whereas Bax reached its maximal expression after 4 weeks. Subsequently, the expression of Bcl-2 and Bax gradually decreased. The ratio of Bcl-2/Bax began to decrease 3 days after FMP and continued to decline until 12 weeks. CONCLUSIONS: FMP with functional appliances could change the expression of Bcl-2 and Bax, which is related to apoptosis in condylar chondrocytes. PMID- 18416608 TI - Cranial base features in skeletal Class III patients. AB - OBJECTIVE: To investigate the cranial base configuration in skeletal Class III patients to clarify the conflicting findings from literature. MATERIALS AND METHODS: Initial lateral radiographs of 54 skeletal Class III patients and 54 matched controls (Class I, II/1, II/2) aged 14 to 24 years were analyzed retrospectively for 21 cephalometric basicranial variables and jaw lengths relative to anterior cranial base length. RESULTS: In contrast to overall cranial base length, the anterior (N-S) and posterior (S-Ba, S-Ar) sections failed to show a significant reduction in Class III patients. The significantly more acute angles Ca-S-Ba and Se-S-Ba reflected increased cranial base flexure. Resulting anterior condylar displacement was shown by significant reduction of Se-S-Cd and Ar-Ca. Relative mandibular length was significantly increased. CONCLUSIONS: Decreased basicranial angulation associated with Class III mandibular protrusion was clearly confirmed for skeletal Class III patients. Overall shortening of the cranial base apparently resulted from various minor alterations. The results are compatible with the deficient orthocephalization hypothesis of Class III morphogenesis. The basicranial-maxillary relationship in skeletal Class III remains unclear. PMID- 18416609 TI - Preapplication of orthodontic forces to the donor teeth affects periodontal healing of transplanted teeth. AB - OBJECTIVE: To investigate how the preapplication of orthodontic forces to the donor teeth affects the periodontal healing after transplantation. MATERIALS AND METHODS: The orthodontic force (1.5 cN) was applied to the maxillary right molars of 6-week-old male Spraque-Dawley rats (n = 21) in the experimental side, and the left side of the same animals was used as the control. After 7 days, both right and left maxillary second molars were extracted or replanted. Periodontal conditions were evaluated in the histological specimens 7 days after applying orthodontic force (before and after extraction) and 14 days after replantation. RESULTS: The application of orthodontic force for 7 days significantly increased the periodontal ligament (PDL) space and also the width of the alveolar socket, which resulted in a rich attached PDL to the root surface of the extracted teeth. Significantly more root resorption was also detected in the control side without preapplication of orthodontic force 14 days after replantation. This root resorption might involve in the disruption of the PDL. CONCLUSION: These results suggested that the preapplication of orthodontic force to the donor teeth increased the PDL width and eased the extraction, which might decrease root resorption after replantation. PMID- 18416610 TI - Inconsistencies in the differential diagnosis of open bite. AB - OBJECTIVE: To examine inconsistencies in the differential diagnosis of open bite. MATERIALS AND METHODS: Using visual judgments, a total of 77 anterior open-bite cases in the postpubertal growth period were grouped as either morphogenetic, functional, or combination. The same sample was also grouped as either hyperdivergent, normodivergent, or hypodivergent using mandibular-plane angle and conventional cephalometry. Kappa analysis was used to test the agreement between the 2 methods of evaluation, and chi2 tests were used to analyze the distribution of cephalometrically grouped hyperdivergent, normodivergent, and hypodivergent cases among the visually assessed morphogenetic, functional and combination groups and vice versa. A kappa score of 0.343 indicated a weak agreement between visual judgment and cephalometric methods of evaluation (P < .001). RESULTS: Despite the expectation that cases evaluated as hyperdivergent using cephalometry would be visually evaluated as morphogenetic, more than half of the cases assessed as hyperdivergent were in fact classified as functional. CONCLUSIONS: These findings highlight the inadequacy of relying solely on cephalometric evaluation to classify open bite. PMID- 18416611 TI - Asymmetry of the face in orthodontic patients. AB - OBJECTIVE: To investigate the laterality of the normal asymmetry of the human face, examining differences in laterality in relation to sex, growth stage, and skeletal classification. MATERIALS AND METHODS: A total of 1800 Japanese subjects (651 males and 1149 females; mean age, 15 years 3 months; range, 4 years 2 months to 59 years 11 months) were selected. Individuals in the sample were categorized according to sex, one of three growth stages, and one of three skeletal patterns. Differences in length between distances from the points at which ear rods were inserted to the facial midline and the perpendicular distance from the soft tissue menton to the facial midline were measured on a frontal facial photograph. Subjects with a discrepancy of more than 3 standard deviations of the measurement error were categorized as having left- or right-sided laterality. RESULTS: Of subjects with facial asymmetry, 79.7% had a wider right hemiface, and 79.3% of those with chin deviation had left-sided laterality. These tendencies were independent of sex, age, or skeletal jaw relationships. In this regard, during pubertal growth, the proportion of subjects with wider right hemiface decreased (P < .0001), whereas the proportion of those with a wider left hemiface increased (P < .01), despite a consistent tendency for right-sided dominance. CONCLUSION: These results suggest that laterality in the normal asymmetry of the face, which is consistently found in humans, is likely to be a hereditary rather than an acquired trait. PMID- 18416612 TI - Polymerization efficiency of glass-ionomer and resin adhesives under molar bands. AB - OBJECTIVE: To determine the degree of cure of a light-cured resin-modified glass ionomer (RMGI) under molar bands compared with a light-cured resin and a dual cured resin. MATERIALS AND METHODS: The 3 cements used were Fuji Ortho LC, Eagle Spectrum resin, and Variolink II dual-cure. Each sample was indirectly light cured for 20 seconds (10 seconds occlusally, 10 seconds cervically) under sections of molar bands, and the degree of cure was evaluated with micro-MIR FTIR spectroscopy. RESULTS: The RMGI exhibited a significantly higher mean degree of cure (55.31%) than both of the resins (Eagle 19.23%; Variolink II, 25.42%), which did not differ significantly at alpha = .05 level of significance. CONCLUSION: Higher degree of conversion can be obtained from RMGIs under molar bands compared with composite resin adhesives provided the proper curing technique is used. PMID- 18416613 TI - Dental arch dimensions in Class II division 1 malocclusions with mandibular deficiency. AB - OBJECTIVE: To test the hypothesis that there is no difference in the dimensions of the upper and lower dental arches in Class II division 1 malocclusion with a mandibular deficiency compared to normal Class I occlusion dental arches. MATERIALS AND METHODS: Photocopies of the dental arches of 48 patients exhibiting Class II division 1 malocclusion with mandibular deficiency and of 51 individuals with normal occlusion were compared. Mandibular deficiency was diagnosed clinically. All 99 individuals were in the permanent dentition. The ages of the subjects ranged from 11 years 4 months to 20 years (mean age = 12 years 5 months). RESULTS: When compared to subjects with normal occlusion, the upper dental arches of the Class II division 1 patients presented reduced transverse dimensions and longer sagittal dimensions while the lower arches were less influenced. CONCLUSION: The hypothesis is rejected. Significant differences are present between the dimensions of the upper and lower dental arches in Class II division 1 malocclusion (with a mandibular deficiency and in the permanent dentition) compared to normal Class I occlusion dental arches. PMID- 18416614 TI - Rebonding of orthodontic brackets. Part I, a laboratory and clinical study. AB - OBJECTIVE: To compare rebonding of orthodontic brackets based on the hypothesis that no difference would be found between the adhesive systems with respect to shear bond strength, mode of failure, and clinical failure rates. MATERIALS AND METHODS: The three adhesive systems included two self-etch primers (Transbond and M-Bond) and a conventional phosphoric acid etch (Rely-a-Bond). The sample size was 20 premolars for each adhesive system. The shear bond strength was tested 24 hours after bracket bonding with the bonding/debonding procedures repeated two times after the first debonding. Bond strength, adhesive remnant index (ARI), and failure sites were evaluated for each debonding. Statistical analysis consisted of a two-way analysis of variance (ANOVA) followed by Scheffe analysis. The clinical portion evaluated 15 patients over a 12-month period. RESULTS: The mean shear bond strengths after the first, second, and third debondings for Rely-a Bond were 8.4 +/- 1.8, 10.3 +/- 2.4, and 14.1 +/- 3.3 MPa, respectively; for Transbond 11.1 +/- 4.6, 13.6 +/- 4.5, and 12.9 +/- 4.4 MPa, respectively; and for M-Bond 8.7 +/- 2.7, 10.4 +/- 2.4, and 12.4 +/- 3.4 MPa, respectively. After the three debondings the mean shear bond strength increased significantly from the first to the third debonding for Rely-a-Bond and M-bond (P 99%) biologically active rPA per litre of culture, which can be used for the detection of anthrax and as a potential component of a vaccine against anthrax. PMID- 18416697 TI - Solar power to prevent type 1 diabetes? PMID- 18416698 TI - Assessment and management of hypoglycemia in children and adolescents with diabetes. PMID- 18416699 TI - On the outside looking in: longitudinal muscle development in the gut. PMID- 18416700 TI - Impedance-pH monitoring: new standard for measuring gastro-oesophageal reflux. AB - Impedance monitoring is a new technique that can be used to detect the flow of fluids and gas through hollow viscera. With impedance monitoring gastro oesophageal reflux can be detected independent of its acidity by differences in electrical impedance between the mucosal surface, fluids and gas that surround the catheter. Clinically, it is used in combination with oesophageal pH monitoring, and the combination of impedance-pH monitoring allows recognition of both acidic and weakly acidic reflux episodes. Studies have shown that impedance pH monitoring is useful in the evaluation of patients with proton pump inhibitor resistant typical reflux symptoms, as it provides a higher yield in detecting reflux as the cause of a patient's symptoms compared to pH monitoring alone. It is therefore likely that impedance-pH monitoring will largely replace pH monitoring in the next 5 years and it will become the standard for reflux detection. PMID- 18416702 TI - Fate of foodborne pathogens on green onions and tomatoes by electrolysed water. AB - AIMS: To investigate the efficacy of electrolysed water (EW) in killing Escherichia coli O157:H7, Salmonella typhimurium and Listeria monocytogenes on the surfaces of spot-inoculated green onions and tomatoes. METHODS AND RESULTS: Green onions and tomatoes were inoculated with a cocktail of three strains each of E. coli O157:H7, Salm. typhimurium and L. monocytogenes and treated with acidic electrolysed water (AC-EW), alkaline electrolysed water (AK-EW), alkaline electrolysed water followed by acidic electrolysed water (AK-EW + AC-EW), deionized water followed by acidic electrolysed water (DW + AC-EW) and deionized water (control, DW) for 15 s, 30 s, 1 min, 3 min and 5 min at room temperature (22 +/- 2 degrees C). The relative efficacy of reduction was AC-EW > DW + AC-EW approximately AK-EW + AC-EW > AK-EW > DW. CONCLUSIONS: Acidic EW treatment was able to significantly reduce populations of the three tested pathogens from the surfaces of green onions and tomatoes with increasing exposure time. SIGNIFICANCE AND IMPACT OF THE STUDY: Rinsing in acidic EW reveals an effective method to control the presence of E. coli O157:H7, Salm. typhimurium and L. monocytogenes on the surfaces of fresh green onions and tomatoes, without affecting their organoleptic characteristics. This indicates its potential application for the decontamination of fresh produce surfaces. PMID- 18416703 TI - Molecular characterization of multiple xylanase producing thermophilic/thermotolerant fungi isolated from composting materials. AB - AIMS: Molecular characterization of commercially important group of xylanase producing thermophilic/thermotolerant fungi. METHODS AND RESULTS: DNA from 16 thermophilic/thermotolerant fungal isolates was amplified by PCR using three sets of primers: (i) internal transcribed spacer sequence (ITSI-5.8S-ITSII), (ii) D1/D2 hyper variable region of 26S rDNA and (iii) 18S rDNA region. The amplified products of internal transcribed spacers (ITS) and D1/D2 region were sequenced and analysed using CLUSTALX, whereas, amplified 18S rDNA region was subjected to RFLP analysis based on restriction digestion with RsaI, MboI and Hinf I. CONCLUSIONS: The sequence based analyses of ITSI-5.8S-ITSII as compared with D1/D2 region of 26-28S rDNA was found to be a better tool for phylogenetic resolution of thermophilic/thermotolerant fungi. The ITSI-5.8S-ITSII sequence based dendrogram indicates an early divergence of the alkaline active xylanase producing thermophilic fungal strains. SIGNIFICANCE AND IMPACT OF THE STUDY: This study was the first report on phylogenetic characterization of thermophilic/thermotolerant fungi. PMID- 18416704 TI - Chloraminated drinking water does not generate bacterial resistance to antibiotics in Pseudomonas aeruginosa biofilms. AB - AIM: To determine if exposure of Pseudomonas aeruginosa biofilms to chloraminated drinking water can lead to individual bacteria with resistance to antibiotics. METHODS AND RESULTS: Biofilms of P. aeruginosa PA14 were grown in drinking water in a Kadouri drip-fed reactor; the biofilms were treated with either 0.5 mg l(-1) or 1.0 mg l(-1) of chloramine for 15 or 21 days; control biofilms were grown in water without chloramine. Fewer isolates with antibiotic resistance were obtained from the chloramine-treated biofilms as compared to the control. Minimum inhibitory concentrations (MIC) for selected antibiotic-resistant isolates were determined using ciprofloxacin, tobramycin, gentamicin, rifampicin and chloramphenicol. All of the isolates tested had increased resistance over the wildtype to ciprofloxacin, rifampicin and chloramphenicol, but were not resistant to tobramycin or gentamicin. CONCLUSIONS: Under these test conditions, there was no detectable increase in antibiotic resistance in P. aeruginosa exposed as biofilms to disinfectant residues in chloraminated drinking water. SIGNIFICANCE AND IMPACT OF THE STUDY: Chloramine in drinking water, while unable to kill biofilm bacteria, does not increase the potential of P. aeruginosa to become resistant to antibiotics. PMID- 18416706 TI - Association of maternal HIV infection with increase of infant mortality in Malawi. AB - AIM: To determine the association between maternal HIV infection and infant mortality in Malawi. METHODS: A synthetic cohort life table based on the birth history of 2618 childbirths during 1999 and 2004, from the subsample of 2020 mothers who completed interview and were tested for HIV virus in the 2004 Malawi Demographic and Health Survey was used. The survey collected socio-demographic and health data of a natural representative sample of women aged 15 to 49; and obtained voluntary counselling tests for HIV infection from one-third of the representatives of the sample. Associations of maternal HIV status and other factors with infant mortality were estimated using survival regression analysis and the results are presented as hazard ratios (HR) with level of statistical significance (P-value). RESULTS: Children born to HIV-infected mothers were more than two times as likely to die during infancy as those born to uninfected mothers (HR = 2.21; P < 0.01). Controlling for other risk factors and confounding factors for infant mortality further sharpened this relationship (HR = 2.70; P < 0.01). Boys are more likely to die in infancy than girls. Young mothers and mothers not receiving prenatal care, and low-birthweight children and children living in rural areas, particular so in the northern region, were associated with a higher risk of infant mortality. CONCLUSION: Maternal HIV infection is strongly associated with infant mortality in Malawi independent of many other factors. Results from this study suggest that the HIV/AIDS epidemic has had an enormous impact on child well-being, child survival and infant mortality. The impact increases as the HIV/AIDS epidemic matures and infection in mothers and adults increases. PMID- 18416705 TI - Single versus bihemispheric amplitude-integrated electroencephalography in relation to cerebral injury and outcome in the term encephalopathic infant. AB - BACKGROUND: The demand for early diagnosis and prognostication of cerebral injury in the encephalopathic term infant is increasing to facilitate appropriate management. The single-channel amplitude-integrated electroencephalogram (S-aEEG) has been shown to have predictive utility for the severely encephalopathic infant. New bedside aEEG devices with more channels are entering the neonatal environment. Little data are available to compare the utility of two channels (B aEEG) with that of an S-aEEG recording. AIM: To compare the utility of the S-aEEG and the B-aEEG in the prediction of cerebral injury, as determined by magnetic resonance imaging (MRI), and neurodevelopmental outcome in the term encephalopathic infant. METHODS: Term encephalopathic infants, with or without seizures, admitted to a level III NICU were included in this study. These infants had simultaneous S-aEEG and B-aEEG recordings. MRI was undertaken during the clinical course and classified as to the extent of cerebral injury. Neurological outcome was assessed at 2 years of age. RESULTS: Twenty-eight encephalopathic term infants were included in the study. There was high level of agreement between both brain monitors (Kappa = 0.68, P < 0.001), but there was disagreement in the classification in four cases where the S-aEEG was normal when the B-aEEG was severely abnormal (McNemar's test P = 0.046). Of note in these four cases, all had a severely abnormal MRI and poor neurodevelopmental outcome at 2 years. CONCLUSION: Amplitude measurements by the B-aEEG appear more sensitive in detecting cerebral injury in comparison with the S-aEEG, particularly in the setting of unilateral injury. PMID- 18416707 TI - Ethical issues for parents of extremely premature infants. AB - Evidence suggests that NICU (neonatal intensive care unit) parents with an baby born at the threshold of viability do not always receive sufficient counselling during an emergency admission and as a consequence, are not well-informed to accept withdrawal of treatment or quality of life decisions. As prospective parents are not educated earlier in pregnancy about extreme premature delivery, crucial information and counselling explaining neonatal issues is only offered to labouring women during their emergency admission. As a result, most have difficulty understanding the risks and benefits of baby's treatment and therefore rely heavily on the perinatal physician to take responsibility for the initial treatment. However, this lack of understanding often leaves parents disadvantaged, as many are left unprepared to participate objectively in quality of life decisions. According to recent research, morbidity figures remain relatively high with one in five survivors at risk of a long-term disability. This shows that some parents will still be confronted by ethical decision of whether or not to continue treatment, and this may not be apparent until days after treatment has been established. As recent research has shown, parents do, in fact, want increased involvement in the decision-making process regarding their child's treatment. Therefore, it has been argued, that parents should be provided with information earlier in pregnancy to familiarise themselves with quality of life issues which they may encounter as the NICU parents of an extremely premature infant. PMID- 18416708 TI - Tubulointerstitial nephritis: drugs are not always to blame. PMID- 18416710 TI - A case of Sandifer's syndrome: significance in the differential diagnosis of growth retardation. PMID- 18416709 TI - Long-standing fever and Angelman syndrome: report of two cases. AB - An 8-month-old girl and a 20-month-old boy who presented with motor and developmental delay and long-standing fever are presented. The patients were diagnosed as Angelman syndrome with fluorescence in situ hybridization (FISH) analysis. Despite extensive clinical and laboratory examinations, no inflammatory or infectious origin for the fever was found. It was considered that the long standing fever observed in these cases was due to hypothalamic dysfunction for thermoregulation. PMID- 18416711 TI - A rare umbilical anomaly: trunk like umbilicus. PMID- 18416712 TI - Geographical variation in the use of intertidal rocky shores by the lizard Microlophus atacamensis in relation to changes in terrestrial productivity along the Atacama Desert coast. AB - 1. The movement of materials and organisms between ecosystems is a common process in nature. 2. In the present study we investigate the hypothesis that the movement of consumers between ecosystems depends not only on the differences in productivity between ecosystems and prey availability, but also on these animals' biological characteristics. 3. To address this hypothesis we investigated the changes in abundance, habitat utilization and diet of the lizard Microlophus atacamensis along its geographical range on the coast of the Atacama Desert. Within this range, intertidal rocky shore communities do not show important variations in their species composition and abundance, but terrestrial communities show a steep gradient of productivity associated with the increase in rainfall from north to south. 4. Our results show that the use of intertidal habitats and the consumption of intertidal prey by M. atacamensis change within its geographical range: in the North, the species uses intertidal areas and behaves as an herbivore consuming mostly algae, whereas in the South it expends most of the time in terrestrial habitats as a carnivore mainly of arthropods. 5. Our study gives new evidence for cross-ecosystem connections created by consumer movement between habitats of contrasting but variable productivity levels. PMID- 18416713 TI - A web resource for the UK's long-term individual-based time-series (LITS) data. PMID- 18416716 TI - Health and social care needs of children and young people. PMID- 18416717 TI - Young people supporting parents with mental health problems: experiences of assessment and support. AB - The explosion of interest in young people as carers over the last decade and a half conceals the fact that there are still no reliable estimates of the number of young people with caregiving responsibilities. This is even more problematic in circumstances where the 'looked after' person has a mental health problem. In this study, we reflect on what can be done to identify, assess and support young people in these circumstances. We draw on selected findings from a study that has been examining the constituents of good assessment practice in work with family carers supporting relatives with mental health problems. The study embraces different carer groups but this paper concentrates on the experiences of young carers at one study site where Barnardo's and partner organisations had developed a joint initiative targeting young people who are looking after parents and relatives with mental health problems. Following a review of the literature about young people as carers, the paper describes how Barnardo's worked to support them through its young carers projects. Based on face-to-face interviews with the young people (N = 10) caring for a mother with mental health problems, the main part of the paper provides an account of how they talk about, make sense of and evaluate the support they have received through this combined initiative. The findings underscore the value of one particular young carers project, and provide clues about what lessons may be transferable to other similar projects. PMID- 18416718 TI - Research priorities: Bless thee! Thou art translated. PMID- 18416719 TI - Behaviour after cerebral lesions in children and adults. 1962. PMID- 18416720 TI - Autism and diagnostic substitution. PMID- 18416721 TI - Acute disseminated encephalomyelitis: where does it start and where does it stop? PMID- 18416724 TI - Pass the torch, please! PMID- 18416725 TI - Immediate and early non-occlusal loading of Straumann implants with a chemically modified surface (SLActive) in the posterior mandible and maxilla: interim results from a prospective multicenter randomized-controlled study. AB - OBJECTIVE: Immediate and early loading of dental implants can simplify treatment and increase overall patient satisfaction. The purpose of this 3-year prospective randomized-controlled multicenter study was to assess the differences in survival rates and bone level changes between immediately and early-loaded implants with a new chemically modified surface (SLActive). This investigation shows interim results obtained after 5 months. MATERIAL AND METHODS: Patients > or =18 years of age missing at least one tooth in the posterior maxilla or mandible were enrolled in the study. Following implant placement, patients received a temporary restoration either on the day of surgery (immediate loading) or 28-34 days after surgery (early loading); restorations consisted of single crowns or two to four unit fixed dental prostheses. Permanent restorations were placed 20-23 weeks following surgery. The primary efficacy variable was change in bone level (assessed by standardized radiographs) from baseline to 5 months; secondary variables included implant survival and success rates. RESULTS: A total of 266 patients were enrolled (118 males and 148 females), and a total of 383 implants were placed (197 and 186 in the immediate and early loading groups, respectively). Mean patient age was 46.3+/-12.8 years. After 5 months, implant survival rates were 98% in the immediate group and 97% in the early group. Mean bone level change from baseline was 0.81+/-0.89 mm in the immediate group and 0.56+/-0.73 mm in the early group (P<0.05). Statistical analysis revealed a significant center effect (P<0.0001) and a significant treatment x center interaction (P=0.008). CONCLUSIONS: The results suggested that Straumann implants with an SLActive can be used predictably in time-critical (early or immediate) loading treatment protocols when appropriate patient selection criteria are observed. The mean bone level changes observed from baseline to 5 months (0.56 and 0.81 mm) corresponded to physiological observations from other studies, i.e., were not clinically significant. The presence of a significant center effect and treatment x center interaction indicated that the differences in bone level changes between the two groups were center dependent. PMID- 18416726 TI - Immediate transmucosal implant placement in molar extraction sites: a 12-month prospective multicenter cohort study. AB - AIM: To assess the clinical and radiographic outcomes of immediate transmucosal placement of implants into molar extraction sockets. STUDY DESIGN: Twelve-month multicenter prospective cohort study. MATERIAL AND METHODS: Following molar extraction, tapered implants with an endosseous diameter of 4.8 mm and a shoulder diameter of 6.5 mm were immediately placed into the sockets. Molars with evidence of acute periapical pathology were excluded. After implant placement and achievement of primary stability, flaps were repositioned and sutured allowing a non-submerged, transmucosal healing. 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 bioresrobable collagen membrane. Standardized radiographs were obtained at baseline and 12 months thereafter. Changes in depth and width of the distance from the implant shoulder (IS) and from the alveolar crest (AC) to the bottom of the defect (BD) were assessed. RESULTS: Eighty-two patients (42 males and 40 females) were enrolled and followed for 12 months. They contributed with 82 tapered implants. Extraction sites displayed sufficient residual bone volume to allow primary stability of all implants. Sixty-four percent of the implants were placed in the areas of 36 and 46. GBR was used in conjunction with the placement of all implants. No post-surgical complications were observed. All implants healed uneventfully yielding a survival rate of 100% and healthy soft tissue conditions after 12 months. Radiographically, statistically significant changes (P<0.0001) in mesial and distal crestal bone levels were observed from baseline to the 12 month follow-up. CONCLUSIONS: The findings of this 12-month prospective cohort study showed that immediate transmucosal implant placement represented a predictable treatment option for the replacement of mandibular and maxillary molars lost due to reasons other than periodontitis including vertical root fractures, endodontic failures and caries. PMID- 18416727 TI - Photo-induced hydrophilicity enhances initial cell behavior and early bone apposition. AB - OBJECTIVE: The anatase form of titanium dioxide (TiO(2)) exhibits photo-induced hydrophilicity when it is irradiated with ultraviolet (UV) light. In the present study, the effect of photo-induced hydrophilicity on initial cell behavior and bone formation was evaluated. MATERIALS AND METHODS: Plasma source ion implantation method and post-annealing were employed for coating the anatase form of TiO(2) to the surface of the titanium disk and implant. Half of the disks and implants were illuminated with UV for 24 h beforehand, whereas the other halves were blinded and used as controls. Photo-induced hydrophilicity was confirmed by a static wettability assay. The effects of this hydrophilicity on cell behavior were evaluated by means of cell attachment, proliferation and morphology using pluripotent mesenchymal precursor C2C12 cells. Thereafter, bone formation around the hydrophilic implant inserted in the rabbit tibia was confirmed histomorphometrically. RESULTS: The water contact angle of the photo-induced hydrophilic disk decreased markedly from 43.5 degrees to 0.5 degree. Cell attachment and proliferation on this hydrophilic disk showed significant improvement. The cell morphology on this hydrophilic disk was extremely flattened, with an elongation of the lamellipodia, whereas a round/spherical morphology was observed on the control disk. The photo-induced hydrophilic implant enhanced the bone formation with the bone-to-metal contact of 28.2% after 2 weeks of healing (control: 17.97%). CONCLUSION: The photo-induced hydrophilic surface used in the current study improves the initial cell reactions and enhances early bone apposition to the implant. PMID- 18416728 TI - Vivosorb, Bio-Gide, and Gore-Tex as barrier membranes in rat mandibular defects: an evaluation by microradiography and micro-CT. AB - OBJECTIVES: The objectives of this study were to determine whether a new degradable synthetic barrier membrane (Vivosorb) composed of poly(dl-lactide epsilon-caprolactone) (PDLLCL) can be useful in implant dentistry and to compare it with collagen and expanded polytetrafluoroethylene (ePTFE) membranes. MATERIAL AND METHODS: In 192 male Sprague-Dawley rats, a standardized 5 mm circular defect was created through the right angle of the mandible. New bone formation was evaluated by post-mortem microradiography and micro-CT (muCT) imaging. Four groups (control, PDLLCL, collagen, ePTFE) were evaluated at three time intervals (2, 4, and 12 weeks). In the membrane groups the defects were covered; in the control group the defects were left uncovered. Data were analysed using a multiple regression model. RESULTS: New bone formation could be detected by post mortem microradiography in 130 samples and by muCT imaging in 112 samples. Bone formation was progressive in 12 weeks, when the mandibular defect was covered with a membrane. Overall, more bone formation was observed underneath the collagen and ePTFE membranes than the PDLLCL membranes. CONCLUSIONS: In contrast to uncovered mandibular defects, substantial bone healing was observed in defects covered with a PDLLCL membrane. However, bone formation in PDLLCL-covered defects tended to be less than in the defects covered with collagen or ePTFE. The high variation in the PDLLCL samples at 12 weeks may be caused by the moderate adherence of this membrane to bone compared with collagen. These results indicate that further study is needed to optimize the properties of PDLLCL membranes. PMID- 18416729 TI - Endoscopic dilatation in Crohn's disease. PMID- 18416731 TI - The AJT Report. News and issues that affect organ and tissue transplantation. PMID- 18416732 TI - A closer look at T cells in the artery wall. PMID- 18416733 TI - Machine perfusion for 'marginal' liver grafts. AB - Due to the critical shortage of deceased donor grafts, clinicians are continually expanding the criteria for an acceptable liver donor to meet the waiting list demands. However, the reduced ischemic tolerance of those extended criteria grafts jeopardizes organ viability during cold storage. Machine perfusion has been developed to limit ischemic liver damage but despite its proven biochemical benefit, machine liver perfusion is not yet considered clinically due to its low practicability. In this review, we summarize our understanding of the role of machine perfusion in marginal liver preservation. The goal is to highlight advantages or disadvantages of current perfusion techniques and to explain the underlying mechanisms. We provide evidence for the need of a liver perfusion performance shortly before implantation, and point out promising designs. PMID- 18416734 TI - Organ trafficking and transplant tourism: a commentary on the global realities. AB - The extent of organ sales from commercial living donors (CLDs) or vendors has now become evident. At the Second Global Consultation on Human Transplantation of the World Health Organization's (WHO) in March 2007, it was estimated that organ trafficking accounts for 5-10% of the kidney transplants performed annually throughout the world. Patients with sufficient resources in need of organs may travel from one country to another to purchase a kidney (or liver) mainly from a poor person. Transplant centers in 'destination' countries have been well known to encourage the sale of organs to 'tourist' recipients from the 'client' countries. PMID- 18416735 TI - First things first: laying the ethical and factual groundwork for living kidney donor selection standards. PMID- 18416736 TI - OPTN policy regarding prioritization of patients with hepatopulmonary syndrome: does it provide equitable organ allocation? AB - United Network for Organ Transplantation (UNOS) policy 3.6.4.5.1 provides exception points to patients diagnosed with hepatopulmonary syndrome (HPS) to compensate for their reported increased mortality risk. We compared pre- and posttransplant and overall outcomes in 255 patients receiving exception points under this policy (HPS policy patients) with 32 358 nonexception control patients listed in the model for end-stage liver disease (MELD) era to determine whether the intent of the policy is being met. Overall, 92.5% of HPS policy patients versus 45.5% of controls had been transplanted, 5.1% versus 31.2% remained on waiting list and 1.5% versus 14.1% had died while awaiting transplant (p < 0.0001 for each comparison). Relative risk (RR) of death for HPS policy patients compared to controls was 0.158 (confidence interval [CI]: 0.059-0.420, p = 0.0002) pretransplant, and 0.827 (CI: 0.587-1.170, p = 0.28) posttransplant. Overall (combined waitlist and posttransplant) RR of death was 0.514 (CI: 0.374 0.707, p = 0.00004) compared with controls. After adjustment for laboratory MELD, overall RR was 0.807 (CI: 0.587-1.110, p = 0.19), indicating that HPS policy patients' mortality risk would be similar to that of controls had they been listed with their laboratory MELD score. HPS policy patients have a significant pretransplant survival advantage over standard liver transplant candidates because of the exception points awarded, and have similar posttransplant survival. Better criteria for diagnosing and grading of HPS are required. PMID- 18416737 TI - Incidence of PTLD in pediatric renal transplant recipients receiving basiliximab, calcineurin inhibitor, sirolimus and steroids. AB - Pediatric renal transplant recipients were enrolled in a multicenter, randomized, double-blind trial of steroid withdrawal. Subjects received basiliximab, calcineurin inhibitor, sirolimus and steroids. Of 274 subjects enrolled, 19 (6.9%) subjects developed posttransplant lymphoproliferative disorder (PTLD). The relative hazard (RH) for PTLD was 5.3-fold higher in children aged < or =5 versus those >12 years (p = 0.0017). EBV seronegative subjects had a 4.7-fold higher RH compared to EBV positive subjects (p = 0.02). Among EBV donor+/recipient- (D+/R-) subjects, the RH increased by 6.1-fold (p = 0.0001). In a multivariate model, risk factors included recipient age < or =5 years (RH 3.2, 95% CI: 1.1-9.6, p = 0.034) and EBV D+/R- status (RH 7.7, 95% CI: 1.6-35.9, p = 0.010). Of 19 patients with PTLD, 17 are alive with functioning grafts and 2 lost their grafts, 1 of whom subsequently died of recurrent PTLD. This 'robust' immunosuppression protocol was associated with low rejection rates but an unacceptably high incidence of PTLD. The combination of basiliximab, calcineurin inhibitor, sirolimus and steroids resulted in over-immunosuppression in a high-risk pediatric population and we do not recommend its use. Future studies must include routine viral monitoring to permit early identification of viral activity and a protocol driven reduction of immunosuppression aimed at avoiding complications. PMID- 18416738 TI - Comparing microarray versus RT-PCR assessment of renal allograft biopsies: similar performance despite different dynamic ranges. AB - In renal allografts, assessing gene expression can add relevant diagnostic information to histopathology. Results can be expressed as single genes or gene sets, representing pathogenesis-based transcript sets (PBTs): cytotoxic T-cell associated, interferon gamma- induced or decreased kidney parenchymal transcripts. Two technology platforms are available: RT-PCR and microarrays. We compared RT-PCR, U133plus2.0 microarrays and histopathology in 86 biopsies. We compared 13 potentially diagnostic genes as RT-PCR probes to microarray-derived PBTs, 'mini'-PBTs (small sets of 3-5 transcripts) and a histology classifier. Most RT-PCR probes (10/13) correlated well with the corresponding microarray probe sets (r > 0.8). Exceptions included FASLG and CD8B1 microarray probe sets, which were not performing on microarrays but were detectable by RT-PCR most likely due to differences in sensitivity. In general, RT-PCR showed greater dynamic range, detecting small changes in normal kidneys, but RT-PCR and microarrays gave similar results in abnormal kidneys. Individual transcripts or mini-PBTs assessed by either platform correlated well with one another, with microarray PBTs and the histology classifier. Thus, microarrays and RT-PCR assessments agree strongly with one another and histopathology in assessing transplant inflammation, particularly, when results are expressed as PBTs or mini PBTs. The dynamic range of both platforms was sufficient to detect the relevant changes in rejection. PMID- 18416739 TI - Malignancy after heart transplantation: incidence, prognosis and risk factors. AB - The Spanish Post-Heart-Transplant Tumour Registry comprises data on neoplasia following heart transplantation (HT) for all Spanish HT patients (1984-2003). This retrospective analysis of 3393 patients investigated the incidence and prognosis of neoplasia, and the influence of antiviral prophylaxis. About 50% of post-HT neoplasias were cutaneous, and 10% lymphomas. The cumulative incidence of skin cancers and other nonlymphoma cancers increased with age at HT and with time post-HT (from respectively 5.2 and 8.9 per 1000 person-years in the first year to 14.8 and 12.6 after 10 years), and was greater among men than women. None of these trends held for lymphomas. Induction therapy other than with IL2R-blockers generally increased the risk of neoplasia except when acyclovir was administered prophylactically during the first 3 months post-HT; prophylactic acyclovir halved the risk of lymphoma, regardless of other therapies. Institution of MMF during the first 3 months post-HT reduced the incidence of skin cancer independently of the effects of sex, age group, pre-HT smoking, use of tacrolimus in the first 3 months, induction treatment and antiviral treatment. Five-year survival rates after first tumor diagnosis were 74% for skin cancer, 20% for lymphoma and 32% for other tumors. PMID- 18416740 TI - T cells in cardiac allograft vasculopathy are skewed to memory Th-1 cells in the presence of a distinct Th-2 population. AB - Cardiac allograft vasculopathy (CAV) in heart transplantation (HTx) patients remains the major complication for long-term survival, due to concentric neointima hyperplasia induced by infiltrating mononuclear cells (MNC). Previously, we showed that activated memory T-helper-1 (Th-1) cells are the major component of infiltrating MNC in coronary arteries with CAV. In this study, a more detailed characterization of the MNC in human coronary arteries with CAV (n = 5) was performed and compared to coronary arteries without CAV (n = 5), by investigating MNC markers (CD1a, DRC-1, CD3, CD20, CD27, CD28, CD56, CD68, CD69, FOXP3 and HLA-DR), cytokines (IL-1A, 2, 4, 10, 12B, IFN-gamma, and TGF-beta1), and chemokine receptors (CCR3, CCR4, CCR5, CCR7, CCR8, CXCR3 and CX3CR1) by immunohistochemical double-labeling and quantitative PCR on mRNA isolated from laser microdissected layers of coronary arteries. T cells in the neointima and adventitia of CAV were skewed toward an activated memory Th-1 phenotype, but in the presence of a distinct Th-2 population. FOXP3 positive T cells were not detected and production of most cytokines was low or absent, except for IFN gamma, and TGF-beta. This typical composition of T-helper cells and especially production of IFN-gamma and TGF-beta may play an important role in the proliferative CAV reaction. PMID- 18416741 TI - Regional and racial disparities in the use of live non-directed kidney donors. AB - Use of live non-directed donors (LNDDs), or altruistic donors, has increased significantly over the past decade and has fueled debate regarding the ethics and allocation of this new source of live donor kidneys. Three allocation philosophies are currently in use, including donor-centric, recipient-centric and socio-centric models, and our group has also advocated the use of LNDDs in paired donation. However, no universally accepted allocation policy exists, nor does national oversight. To determine allocation patterns resulting from current practice models, we analyzed the 372 LNDD kidney transplants performed in the United States since 1998. Most LNDD transplants occurred at a minority of centers, with only five centers performing over 10, and over 28% of LNDDs traveled out-of-state to donate. Furthermore, a center's use of LNDD kidneys did not correlate with that center's organ shortage. Finally, African Americans were significantly under-represented among recipients who were allocated LNDD kidneys, even after accounting for differences in the racial makeup of the waiting list representing centers using LNDD kidneys. These disparities suggest the need for continued monitoring and discussion of LNDD at a national level. If non-directed donation continues to rise at its current rate, a national allocation policy may be reasonable. PMID- 18416743 TI - Heme oxygenase-1 polymorphisms and renal transplantation outcomes: balancing at the detection limit of allelic association studies. PMID- 18416747 TI - Factors influencing efficiency of sliding mechanics to close extraction space: a systematic review. AB - OBJECTIVES: To review recent literature to determine strength of clinical evidence concerning the influence of various factors on the efficiency (rate of tooth movement) of closing extraction spaces using sliding mechanics. DESIGN: A comprehensive systematic review on prospective clinical trials. An electronic search (1966-2006) of several databases limiting the searches to English and using several keywords was performed. Also a hand search of five key journals specifically searching for prospective clinical trials relevant to orthodontic space closure using sliding mechanics was completed. Outcome Measure - Rate of tooth movement. RESULTS: Ten prospective clinical trials comparing rates of closure under different variables and focusing only on sliding mechanics were selected for review. Of these ten trials on rate of closure, two compared arch wire variables, seven compared material variables used to apply force, and one examined bracket variables. Other articles which were not prospective clinical trials on sliding mechanics, but containing relevant information were examined and included as background information. CONCLUSION - The results of clinical research support laboratory results that nickel-titanium coil springs produce a more consistent force and a faster rate of closure when compared with active ligatures as a method of force delivery to close extraction space along a continuous arch wire; however, elastomeric chain produces similar rates of closure when compared with nickel-titanium springs. Clinical and laboratory research suggest little advantage of 200 g nickel-titanium springs over 150 g springs. More clinical research is needed in this area. PMID- 18416748 TI - Short-term changes in temporomandibular joint function in subjects with cleft lip and palate treated with maxillary distraction osteogenesis. AB - OBJECTIVES: To investigate the short-term effects of maxillary distraction osteogenesis (DO) on temporomandibular joint (TMJ) function in 21 subjects with cleft lip and palate (CLP). Design - Morphological changes in the maxillofacial region were measured using lateral cephalometric radiographs taken immediately before (pre-DO) and after DO (post-DO) and 1 year after DO (1-year follow-up). A questionnaire was evaluated using a visual analog scale. A chi-square test was used to compare the prevalence of TMJ symptoms between pre-DO and 1-year follow up. The Spearman correlation coefficient was used to determine the correlation between changes in cephalometric variables and TMJ symptoms in association with maxillary DO. Statistical significance was set at p < 0.05. Results - The ANB (anteroposterior relationship of the maxilla with the mandible) angle and the mandibular plane angle at pre-DO, post-DO, and 1-year follow-up were -4.3 degrees , +5.8 degrees , +4.3 degrees and 32.1 degrees , 33.5 degrees , 33.6 degrees , respectively. The average amounts of anterior and downward movement of the maxilla at post-DO and 1-year follow-up were 8.3, -1.3 and 0.9, 1.1 mm, respectively. The prevalence of TMJ symptoms showed no significant increase in association with maxillary DO. Moreover, there was no significant correlation between changes in cephalometric variables and TMJ symptoms. Conclusion - These results suggest that there was no short-term (i.e., up to 1 year after DO) effect of maxillary DO on TMJ function in subjects with CLP. PMID- 18416749 TI - Spacing and crowding among African and Caucasian children. AB - OBJECTIVE: To determine spacing and crowding according to ethnic group, gender and dental emergence stage among Tanzanian African and Caucasian children. DESIGN: Cross-sectional epidemiological clinical study. SETTING: A total of 869 African (428 boys, 441 girls) and 706 Caucasian (319 boys, 387 girls) school children, aged 3(1/2)-16 years. MAIN OUTCOME MEASURES: Comparison of spacing and crowding between African and Caucasian children according to gender and dental emergence stage. RESULTS: Spacing was more often found in the maxilla, while crowding was more common in the mandible. Only during the transition of the maxillary permanent front teeth was there significantly more spacing in Caucasians. No gender differences were found. In both samples spacing decreased during later emergence stages. Crowding was more often found in Caucasian children than in African children. In Caucasian children the frequency of crowding increased with advanced emergence stages, while for Africans the trend was not consistent. CONCLUSION: When planning resources for orthodontic treatment for different populations as well as planning treatment for individuals, ethnic background and emergence stage of the dentition need to be considered. PMID- 18416750 TI - A new system for scanning, measuring and analyzing dental casts based on a 3D holographic sensor. AB - OBJECTIVES: To evaluate the reliability of a new technique for measuring 3D scanned orthodontic cast models with cross-section planes using teledent, a new software, developed at Technion - for the purpose of this research. EXPERIMENTAL VARIABLE: Thirty orthodontic plaster models were divided into three equal groups according to severity of teeth crowding. Measurements of mesio-distal tooth width and the arch length were performed manually on the casts using a conventional caliper. Thereafter, the models were scanned and processed into the software using a 3-D measuring scanner with a holographic sensor 'ConoProbe' (by Optimet, Jerusalem, Israel). teledent used two types of digital measurements; linear and cross-section planes to perform a space analysis on the scanned teeth. Significance was determined by the paired Wilcoxon rank sum test. RESULTS: Results show that 3D measurements of tooth width and arch length obtained by cross-section planes were generally similar to manual caliper measurements, while linear measurements were statistically smaller. When comparing space analysis, both digital measurements were statistically smaller than the caliper (p < 0.05). The difference in space analysis between the caliper and the cross-section plane measurements was very small (0.38-0.74 mm) and can be considered clinically acceptable. However, a difference of 1.19-3 mm between the linear measurements and the caliper might have clinical implications especially in severely crowded dentition. CONCLUSIONS: This study suggests that measurements performed by cross section planes are as accurate as the manual caliper and can be employed clinically while the accuracy of linear measurements is sometimes questionable. PMID- 18416751 TI - The neurocranial and craniofacial morphology in children with solitary median maxillary central incisor (SMMCI). AB - OBJECTIVES: The purpose of this study was to describe the neurocranial and craniofacial morphology on profile radiographs of children with single median maxillary central incisor (SMMCI). DESIGN: Cephalometric analyses of neurocranium and craniofacial morphology on profile radiographs. SETTING AND SAMPLE POPULATION: Department of Orthodontics, School of Dentistry, University of Copenhagen. Thirteen children with SMMCI, 12 girls and one boy (7-17 years of age). OUTCOME MEASURE: Cephalometric measurement were compared with normal standards using a paired t-test and Wilcoxon signed rank test. RESULTS: The size of the neurocranium (especially the length of the anterior cranial base), the maxillary prognathia, the maxillary inclination, the mandibular prognathia and the inclination of the mandibular incisors are significantly reduced in SMMCI. The mandibular inclination, the vertical jaw relationship, the alveolar bone prognathia in the upper jaw and the mandibular angle are significantly enlarged in SMMCI. CONCLUSION: The present study showed that occurrence of SMMCI is a sign of a developmental anomaly associated with deviations in neurocranial size and shape and in craniofacial morphology. PMID- 18416752 TI - Histopathological changes in collagen and matrix metalloproteinase levels in articular condyle of experimental model rats with jaw deformity. AB - OBJECTIVE: To investigate the dynamics of the cartilage matrix in the articular condyle after removal of a side shift plate; Emergence of type I, II, and III collagen in the matrix as well as changes in levels of matrix metalloproteinase (MMP)-1, -8, and -13 that degrade collagen were studied histopathologically and immunohistochemically. DESIGN: Lateral displacement of the mandible was achieved by attaching a side shift plate to the anterior teeth of the maxilla in male rats at 6 weeks. The wearing period of the side shift plate was 8 weeks. Observations were made at 0, 1, 2, 4 and 8 weeks after removal. RESULTS: In histopathological findings, the timing of proliferation of the layer of hypertrophy varied between the bilateral sides. In immunohistochemical findings a significant decline in the expression of type II collagen in the displacement side was observed immediately after removal. Moreover, the expressions of MMPs were elevated in both sides on 0 weeks. At 1 week after removal, a significant elevated in the expression of type II collagen, MMPs was decline in both sides. CONCLUSIONS: After removal, the levels of MMP-1, -8, and 13 were reduced and the emergence of type II collagen increased. Thus, cellular outgrowth was initiated to trigger intracartilaginous ossification to restore the cartilage matrix. PMID- 18416753 TI - Skull thickness in patients with skeletal deep bite. AB - OBJECTIVES: Skull thickness in relation to patients with skeletal deep bite has not been reported previously. The present study examines the skull thickness in patients with skeletal deep bite and compares it with the skull thickness in subjects with neutral occlusion and normal craniofacial morphology. DESIGN: A retrospective case-control study. SETTING AND SAMPLE POPULATION: The material comprised 36 patients divided into two groups, a group of 18 patients with skeletal deep bite (eight females, 10 males) and a control group of 18 subjects (eight females, 10 males) with neutral occlusion and normal craniofacial morphology. OUTCOME: Skull thickness measurements. RESULTS: No significant gender differences were found regarding skull thickness. The skull was thicker in the deep bite group compared with the group with neutral occlusion and normal craniofacial morphology (p < 0.05). CONCLUSION: The present study demonstrates that patients with skeletal deep bite have a significantly thicker skull than subjects with neutral occlusion and normal vertical craniofacial morphology. This is considered important for the treatment planning for orthodontic and orthognathic patients. PMID- 18416754 TI - Lipids and skin barrier function--a clinical perspective. AB - The stratum corneum (SC) protects us from dehydration and external dangers. Much is known about the morphology of the SC and penetration of drugs through it, but the data are mainly derived from in vitro and animal experiments. In contrast, only a few studies have the human SC lipids as their focus and in particular, the role of barrier function in the pathogenesis of skin disease and its subsequent treatment protocols. The 3 major lipids in the SC of importance are ceramides, free fatty acids, and cholesterol. Human studies comparing levels of the major SC lipids in patients with atopic dermatitis and healthy controls have suggested a possible role for ceramide 1 and to some extent ceramide 3 in the pathogenesis of the disease. Therapies used in diseases involving barrier disruption have been sparely investigated from a lipid perspective. It has been suggested that ultraviolet light as a treatment increases the amount of all 3 major SC lipids, while topical glucocorticoids may lead to a decrease. Such effects may influence the clinical outcome of treatment in diseases with impaired barrier function. We have, therefore, conducted a review of the literature on SC lipids from a clinical perspective. It may be concluded that the number of human studies is very limited, and in the perspective of how important diseases of impaired barrier function are in dermatology, further research is needed. PMID- 18416755 TI - Polymorphisms in the interleukin-1 gene influence the stratum corneum interleukin 1 alpha concentration in uninvolved skin of patients with chronic irritant contact dermatitis. AB - BACKGROUND: Interleukin (IL)-1 alpha and its receptor antagonist IL-1 ra play a role in skin inflammation. Several polymorphisms in the IL1 gene cluster, coding for IL-1 alpha, IL-1 ra, and IL-1 beta, influence their protein expression. Within this cluster, strong linkage disequilibrium has been shown. OBJECTIVE: We studied the association between the polymorphisms IL1A-889 (C-->T) and IL1B-31 (T ->C) and the concentration of IL-1 alpha and IL-1 ra in the stratum corneum (SC). METHOD: In 124 patients with chronic irritant contact dermatitis, we genotyped the IL1A-889 and IL1B-31 polymorphisms and determined the amount of IL-1 alpha and IL-1 ra on tape strips obtained from uninvolved skin of the volar forearm. RESULTS: The SC IL-1 alpha concentration was 23% and 47% lower in subjects with IL1A-889 C/T genotype and T/T genotype, respectively, compared with wild-type genotype. In subjects with IL1B-31 C/C genotype, the IL-1 alpha concentration was 51% lower compared with C/T and T/T genotypes. The ratio IL-1 ra/IL-1 alpha increased twofold in IL1A-889 C/T genotype and threefold in T/T genotype compared with wild type. CONCLUSIONS: We have shown a clear effect of IL1 genotype on protein expression in the SC. This altered expression may be responsible for the interindividual differences in the inflammatory response of the skin. PMID- 18416756 TI - Cytokine gene polymorphisms and susceptibility to chronic irritant contact dermatitis. AB - BACKGROUND: Cytokines play an important role in skin inflammation. OBJECTIVES: We determined whether polymorphisms in cytokine genes contribute to the occurrence of occupational chronic irritant contact dermatitis (CICD). METHODS: In a case control study, 9 polymorphisms in the genes coding for interleukin (IL)-1 alpha, IL-1 beta, IL-8, IL-10, and tumour necrosis factor (TNF)-alpha were determined in 197 patients with CICD. 217 apprentices in vocational training for high-risk occupations for CICD served as controls. RESULTS: For all polymorphisms, no differences in genotype distributions were found between patients and controls. However, in patients with self-reported low levels of wet work and irritant exposure, more TNFA -308 variant genotypes (G/A and A/A) were present compared with those exposed to higher levels or controls, which indicates a TNFA-induced increase of susceptibility. In patients with TNFA -308 variant genotypes, the prevalence of flexural eczema was higher (48% and 57%) compared with that in patients presented with wild-type genotype (30%). Regarding IL1A -889, prevalence of symptoms of dermatitis was lower in apprentices with T/T or C/T genotype (32% and 36%) compared with wild-type genotype (54%, C/C). This indicates a protective effect of these variant alleles in acquiring hand dermatitis. CONCLUSIONS: This study provides evidence that some genetic variations alter susceptibility to (chronic) dermatitis. Knowledge of the impact of genetic differences on the risk of CICD is essential in predictive testing of individuals at risk. PMID- 18416757 TI - Significant exposures to isoeugenol derivatives in perfumes. AB - BACKGROUND: Isoeugenol, an important fragrance allergen in consumers, has been restricted to 200 p.p.m. since 1998 according to guidelines issued by the fragrance industry. However, no decline in contact allergy to isoeugnol has been detected. It has been speculated that isoeugenol derivatives, especially isoeugenyl acetate, are used instead. Isoeugenyl acetate is probably metabolized in the skin to isoeugenol and gives positive patch test reactions in 1/3 of isoeugenol-sensitized individuals. OBJECTIVES: To investigate the content of isoeugenol, isoeugenyl acetate, and two isoeugenol ethers in perfumes/aftershaves. MATERIALS AND METHODS: 29 international brand perfumes/aftershaves were analysed for the target fragrance ingredient by gas chromatography-mass spectrometry. All samples were analysed in duplicate at detection levels of 1-5 p.p.m. RESULTS: 16 products (55%) contained isoeugenol. The maximum concentration was 202 p.p.m. 10 products (34%) contained isoeugenyl acetate, which in 9 cases occurred together with isoeugenol. The concentrations of isoeugenyl acetate ranged from 20 to 4689 p.p.m. 13 products (44%) contained 64.9-1755.0 p.p.m. isoeugenyl methyl ether. Isoeugenyl benzyl ether was not detected in any of the investigated products. CONCLUSIONS: Isoeugenyl acetate is present in perfumes/aftershaves, in some products in significant amounts. This may lead to elicitation of contact allergy in isoeugenol-sensitized individuals and may contribute to unchanged levels of isoeugenol sensitization. PMID- 18416758 TI - Occupational contact dermatitis due to essential oils. AB - BACKGROUND: Allergic contact dermatitis induced by the occupational use of products containing essential oils has not been studied comprehensively. OBJECTIVES: The aim of the present report was to describe the characteristics, diagnosis, and outcome of 5 patients with occupational contact dermatitis because of essential oils attending our outpatient dermatology clinics over a 2-year period. PATIENTS: These patients are added to the 11 cases reported thus far in the literature. RESULTS: The research shows that for proper diagnosis, patch tests with the standard series and the fragrance series should be performed, in addition to tests with the specific oils to which the patients were exposed. CONCLUSIONS: Patients should be instructed to avoid the allergens identified. Sensitization to essential oils has important implications for the occupational future of affected individuals. PMID- 18416759 TI - Occupational dermatoses in health care workers evaluated for suspected allergic contact dermatitis. AB - BACKGROUND: Occupational skin diseases, including allergic contact dermatitis (ACD), irritant contact dermatitis, and allergic contact urticaria (ACU), occur commonly among health care workers (HCWs). PURPOSE: To evaluate the aetiology of the various skin diseases afflicting HCWs evaluated for suspicion of ACD and/or ACU and to identify the most common allergens among HCWs found to have ACD and/or ACU. METHODS: A total of 1434 patients underwent patch testing. The demographic data and most common allergens for HCWs (n = 100) and non-HCWS (n = 1334) were compared. RESULTS: HCWs were statistically more likely than non-HCWS to be female, have hand dermatitis, and have a history of atopy. HCWs were also more likely to have work-related ACD especially to quaternium-15, thiuram, carba mix, glutaraldehyde and benzalkonium chloride, and to have ACU to latex. LIMITATIONS: This study was retrospective and is subject to the resultant biases of all such investigations. Only patients suspected of having ACD and who underwent patch testing are included in our database. The prevalence of ACD and ACU is likely to be higher than that seen in the general HCW population. CONCLUSIONS: Our results underscore the importance of thoroughly evaluating HCWs for ACD and ACU with the use of expanded standard allergen series and prick or radioallergosorbent testing to latex. PMID- 18416760 TI - Reduced frequency of atopic dermatitis in quinoline-allergic patients: the 'hapten-atopy hypothesis'. AB - BACKGROUND: While allergy to food proteins is almost exclusively found in association with atopy, it has been our experience that contact allergy to some contact allergens/haptens with both cutaneous and gastrointestinal exposures is reduced in atopic dermatitis (AD) patients as a group. OBJECTIVE: To assess the contact allergy rates of two classes of antimicrobial haptens, one with both cutaneous and gastrointestinal exposures (quinolines) and one with only significant cutaneous exposure (aminoglycosides), with respect to the presence or absence of AD. METHODS: Contact allergy rates to neomycin (aminoglycoside) and quinoline mix/clioquinol in patients attending the St John's Institute of Dermatology for diagnostic patch testing were retrospectively analysed; current AD and history of AD were noted. RESULTS: In comparison to neomycin-allergic subjects, there was a highly significant negative association between quinoline contact allergy and current presence of AD (P = 0.0028); negative association between quinoline contact allergy and a history of AD did not reach significance (P = 0.07). CONCLUSIONS: In comparison to an antimicrobial with no significant gastrointestinal exposure (neomycin), contact allergy to quinolines is negatively associated with the presence of AD. This is in contrast to food protein allergy, which is strongly associated with atopy. Possible explanations could include (i) confounding factors or (ii) AD patients are efficient at orally tolerizing haptens and inefficient at orally tolerizing proteins, secondary to their atopic status or (iii) oral tolerance of haptens antagonizes tolerance of food proteins and also leads to an immunological shift towards atopy (hapten-atopy hypothesis). PMID- 18416761 TI - Patients with vulval pruritus: patch test results. AB - BACKGROUND: Persistent vulval pruritus is a frequent problem and patients may not show signs of a primary vulval dermatosis. Allergic contact sensitivity is an important factor in such patients but may also occur as a secondary event in women with vulval dermatoses. OBJECTIVES: We aim to describe women with vulval pruritus who were evaluated for vulval dermatoses. We patch tested to help define or exclude allergens. METHODS: In a 3-year period, 50 women with vulval pruritus were examined for existence of vulval dermatosis. All the patients were patch tested to the European Baseline Series, medicaments, preservatives, vehicles, cosmetics and products they routinely used. RESULTS: Ages of the patients ranged from 22 to 62 years (mean 39.9 +/- 9 years) and duration of the diseases ranged from 0.5-360 months (mean 56.1 +/- 65.6 months). 52% of the women had at least 1 positive patch test. 8 patients (16%) had 1 or more relevant allergic positive reactions. The relevant allergens were usually cosmetics, preservatives and medicaments. CONCLUSIONS: Women with vulval pruritus were patch tested to help define or rule our allergens. Contact allergy incidence in these patients is high. The relevant allergens were usually cosmetics, preservatives and medicaments. PMID- 18416762 TI - Tacrolimus ointment in the treatment of occupationally induced chronic hand dermatitis. AB - BACKGROUND: Occupational hand dermatitis (OHD) is a common and often chronic condition. Usage of topical corticosteroids is limited in long-term therapy because of potential side-effects. New treatment options are highly appreciated. OBJECTIVE: To study efficacy and safety of tacrolimus in patients suffering from OHD. PATIENTS AND METHODS: In a prospective, open, multicentre study, 29 patients with OHD of predominantly mild-to-moderate severity were treated with tacrolimus ointment (Protopic 0.1%) during a 4-week active treatment period with twice daily application followed by a 2-month optional treatment period. Efficacy was evaluated by the use of a standardized hand eczema score and by clinical severity rating, conducted on all study visits (screening, baseline and weeks 2, 4, 6, and 12). Adverse events were recorded for safety evaluation. RESULTS: The hand eczema score declined significantly after 2 weeks of treatment compared with baseline and further declined until the end of the study. Finally, 12 (44%) patients were clear of hand eczema. Worsening of the dermatitis occurred in 2 patients (7%). Clinical grading indicated improvement too. Overall tolerability was good. CONCLUSION: Topical tacrolimus might be an efficacious treatment option for chronic OHD. Blinded and randomized controlled studies are necessary to confirm the results of this pilot study. PMID- 18416763 TI - Anaphylaxis caused by contact urticaria because of epoxy resins: an extraordinary emergency. PMID- 18416764 TI - Allergic contact dermatitis because of Cineraria hybrid in a keen gardener housewife. PMID- 18416766 TI - Tosylamide/formaldehyde resin allergy--a consideration in the atopic toddler. PMID- 18416765 TI - Allergic contact dermatitis due to transdermal buprenorphine. PMID- 18416767 TI - Allergic contact dermatitis to DMDI in an office application. AB - Contact dermatitis to isocyanates is a problem and is usually described within industrial settings. A patient who developed an acute allergic contact dermatitis using a dicyclohexylmethane-4,4'-diisocyanate (DMDI)-charged cartridge to create resin-coated '3D labels' within an office environment is described. PMID- 18416768 TI - Airborne allergic contact dermatitis from 4-(bromomethyl)benzoic acid in a university chemist. PMID- 18416769 TI - Allergic contact dermatitis and asthma caused by limonene in a labourer handling citrus fruits. PMID- 18416770 TI - Allergic contact dermatitis to iodopropynyl butylcarbamate found in a cosmetic cleansing wipe. PMID- 18416772 TI - Addendum to Geier J, Lessmann H, Uter W, Schnuch A. Are concomitant patch test reactions to epoxy resin and BIS-GMA indicative of cross reactivity? Contact Dermatitis 2007: 57: 376-380. PMID- 18416773 TI - Validity of reaction index and positivity ratio? PMID- 18416774 TI - HLA-DRB1 alleles influence clinical phenotypes in Japanese patients with ulcerative colitis. AB - The human leukocyte antigen (HLA) region has been implicated in the disease susceptibility of inflammatory bowel disease by several linkage and association studies. In Caucasians, HLA-DRB1 has been reported to determine the clinical phenotypes of ulcerative colitis (UC). Others and we previously reported that HLA DRB1*1502 was strongly associated with UC in the Japanese population. However, the contribution of HLA-DRB1 to the clinical phenotypes in Japanese UC has not been elucidated yet. The aim of this study was to determine whether HLA-DRB1 alleles were associated with the clinical phenotypes in Japanese patients with UC. A total of 353 patients with UC were recruited. Patients were classified into subgroups by sex, age at diagnosis, disease extent, need for steroid therapy or need for surgical treatment. The allele frequency of HLA-DRB1*08 was significantly higher in patients whose disease extended beyond the rectum (left sided and extensive UC) than in those with proctitis [odds ratio (OR)=2.20, Pc=0.043). The allele frequency of HLA-DRB1*09 was significantly higher in patients with UC diagnosed at the age of 40 years or older than in those with UC diagnosed before the age of 40 years (OR=2.31, Pc=0.022). Besides these positive associations, no significant differences were found in the allele frequencies between the other subgroups. We conclude that HLA-DRB1*09 is associated with the age at diagnosis and HLA-DRB1*08 is associated with the disease extent of UC in Japanese. These results indicate that HLA-DRB1 is not only associated with the overall UC susceptibility but also associated with the clinical phenotypes in Japanese. PMID- 18416775 TI - HNA-1a, HNA-1b and HNA-1c gene frequencies in Argentineans. AB - Human neutrophil antigens (HNA) are polymorphic structures located in the neutrophil membrane. The neutrophil-specific antigens HNA-1a (NA1), 1b (NA2) and 1c (SH) are well-recognized allotypic forms of FcgammaRIIIb and the most frequent targets of neutrophil alloantibodies. The aim of this study was to determine the gene frequencies of the neutrophil-specific antigens belonging to the HNA-1 system in blood donors and Toba Amerindians from Rosario, Argentina. Two hundred and eighteen unrelated healthy Argentinean blood donors and Toba Amerindians from Rosario were typed for HNA-1a, HNA-1b and HNA-1c using polymerase chain reaction with sequence-specific primers. For the Argentinean blood donors, the HNA-1a and HNA-1b gene frequencies were 0.44 and 0.56 and for the Amerindians Toba were 0.77 and 0.23, respectively. The HNA-1c antigen is present in 4.7% (gene frequency=0.023) of the blood donors but in none of the Amerindian individuals. The present data showed that the HNA-1 allele frequencies in the major population and the Toba Amerindians from Rosario are similar to those described in European and others distant Amerindians populations, respectively. PMID- 18416777 TI - What becomes of the whistleblowers? PMID- 18416778 TI - Nursing competence 10 years on: fit for practice and purpose yet? AB - AIMS AND OBJECTIVES: This paper examines how nurses are prepared to be clinically competent and safe at registration, so that they are fit for practice and purpose. It follows up two papers on competence published in 1997 and 1998 and investigates subsequent developments. BACKGROUND: In 1979, major changes in nursing affected nurse education and preparation for competence. In the following two decades, it became clear that nurses lacked clinical skills. This paper examines subsequent changes and asks the question whether this crucial shortcoming has now been remedied. This paper considers the background and context of change in nursing and nurse education in the 1980s. It looks at the new ideology, to prepare the 'knowledgeable doer' and examines the consequences of the change on nursing competency from the 1990s to the present day. METHODS: This is a position paper. Professional policy documents from the English National Board for Nursing, Midwifery and Health Visiting, United Kingdom Central Council for Nursing, Midwifery and Health Visiting and Nursing and Midwifery Council, government reports and legislation on nursing and relevant nursing literature are examined and critically analysed and conclusions drawn. CONCLUSIONS: From 1923 1977, mandatory nursing syllabuses set by the General Nursing Council of England and Wales required the registered nurse to have acquired certain specific clinical skills. These were rigorously tested to an explicit standard set by the General Nursing Council before a nurse was awarded state registration. Twenty five years later, the loss of this system for ensuring this competence and the implications of this loss, have been widely recognised. As a result, many nurse training institutions have introduced clinical skills laboratories, simulation of practice and the Objective Structured Clinical Examination. However, to the authors' surprise and contrary to their initial expectations, the Nursing and Midwifery Council has not made these systems uniform or mandatory and so still has no way of ensuring all nurse training is producing safe nurses in the United Kingdom. The authors conclude that the untested educational ideology that brought root and branch change to nurse training in 1983 and which failed to produce nurses 'fit for practice and purpose' may still prevail. RELEVANCE TO CLINICAL PRACTICE: The present paper demonstrates that United Kingdom nurse training still has no uniform and mandatory system in place to ensure, as far as is possible, that all registered nurses are clinically competent and safe to practice. PMID- 18416779 TI - Engaging service users in the development of stroke services: an action research study. AB - AIMS AND OBJECTIVES: To develop local stroke services by involving, in a meaningful way, those affected by stroke in identifying and prioritising service development issues. BACKGROUND: Consideration should be given to including the views of service users in the evaluation and re-design of services. In stroke, the involvement of patients and carers in the development of services has generally been at a consultative level. There is a need to explore strategies to engage patients and carers more fully with the service development agenda. DESIGN: Action research. METHODS: Within the Reflection Phase of this action research study patients' and carers' experiences of stroke services were explored by semi-structured interviews and focus groups. In the Planning and Action Phases, externally facilitated workgroups comprising patients, carers and professionals used a synthesis of the data to identify a range of service development priorities, which informed the development of action plans. RESULTS: Information provision, preparation for transfer of care and the integration of social and leisure activities appear to be priorities for the development of stroke services from the perspective of patients and carers. These findings informed the work of four collaborative workgroups to lead developments in the stroke pathway as follows: stroke prevention and immediate care, early and continuing rehabilitation, transfer of care and long-term support and information provision. CONCLUSION: Stroke patients and carers played a key and meaningful part in the identification and prioritisation of service development issues. External facilitation ensured progress with service development and maintaining patient and carer participation. Several pragmatic recommendations for enhancing the local delivery of stroke services were made. RELEVANCE TO CLINICAL PRACTICE: With external facilitation it is possible for stroke service users to take a meaningful role in service developments that moves beyond consultation. PMID- 18416780 TI - Shifting roles in nursing--does role extension require role abdication? AB - AIMS AND OBJECTIVES: To discuss findings relating to role extension and loss of nursing care to auxiliary nurses. BACKGROUND: There is ongoing discussion in the literature about what nurses' roles may be and how the extension of role affects patient care. Various models have been devised to measure outcomes and the value of nursing to patients. However, there are limited data on the views of nurses themselves in terms of what they perceive their role to be and what they feel about role change. DESIGN: A qualitative approach was used with the help of the elements of grounded theory. METHODS: Data were analysed using a constant comparative method with core categories identified. The study described in this paper was the final of three. The first two studies involved student nurses and through theoretical sampling, the third sample was chosen to expand the data gained from the students. RESULTS: The findings from the student studies indicated concern that the nursing role was being undertaken by auxiliary nurses. The results of the final study, as discussed in this paper, confirmed this. However, one main difference was that qualified nurses were not necessarily unhappy about auxiliary nurses' role expansion but were concerned that the role of the nurse was moving away from the bedside. CONCLUSIONS: The data suggest that nurses' roles may be hard to define. An abdication of role, as opposed to delegation of role, seems to be occurring. Critical thinking is needed to ensure this is a decision advocated by clinical nurses. RELEVANCE TO CLINICAL PRACTICE: Nurses need to be explicit about what their clinical roles are. This study provides data expressing the views of clinical nurses about role expansion and role abdication and corresponding feelings about it. PMID- 18416781 TI - Just how male are male nurses..? AB - AIM: The aim of the present research study was to elucidate, quantitatively, the gender role perceptions of male nurses in Ireland. BACKGROUND: Caring, women and the female gender role are all historically and fiercely synonymous. However, not all carers are women. For instance, male nurses also assume caring roles. What we do not know is how these men actually relate to their own gender role. Is it possible that because of their immersion in a stereotypically caring career they actually occupy the female gender role? DESIGN: A quantitative non-experimental descriptive design was adopted. METHOD: Short-form Bem sex role inventory was mailed to a random sample of 250 male registered general nurses in Ireland to ascertain whether they perceived themselves to occupy the male or female gender role. RESULTS: One hundred and four men completed the inventory. Overall, the sample identified with more female than male gender norms. Specifically, 78 respondents identified themselves as adhering to more female gender role norms than male gender role norms, whereas 21 respondents identified more strongly with male gender role norms. Five respondents identified equally with both gender roles. CONCLUSION: This study quantitatively elucidates the gender perceptions of male nurses in Ireland for the first time. RELEVANCE TO CLINICAL PRACTICE: Adherence to the female gender role may be an important prerequisite to caring. If this is true, then this study supports the notion that many male nurses occupy this gender role. However, adoption of facets of the female gender role may not be unique to male nurses. Many men may occupy this role and perhaps resultantly be attracted to or well-suited to caring careers. Attracting such men may help in solving the recruitment and retention issues that surround caring careers. Furthermore, the attraction of more men to caring careers may subside the stigma for the minority of men already in such careers. PMID- 18416782 TI - Reliability and validity of the CCNCS: a dependency workload measurement system. AB - AIM: To refine, test and evaluate the Community Client Need Classification System (CCNCS). BACKGROUND: Workload assessment in community nursing is complicated by the range of services that may be delivered in one patient interaction. The CCNCS is a workload measurement system designed to capture the direct and indirect elements of community nursing work and is suitable for use with all care groups in the community. DESIGN: Survey. METHOD: Forty-four community nurses implemented the CCNCS with all clients in their caseload for four weeks. Community nursing in the Irish Republic is known as public health nursing. The Public Health Nurses (PHNs) recorded the total time in minutes that was spent on each client each week. The satisfaction with and experiences of PHNs using the CCNCS during the study period was also recorded. RESULTS: Participants endorsed the utility of the CCNCS for use in community nursing. Inter-rater and intra-rater reliability results were positive with high level of agreement between raters in relation to scoring community clients. The amount of time the PHNs spent with clients correlated with ascending level of client need. CONCLUSIONS: The CCNCS affords insight into the complex nature of community nursing. It discriminates between levels of need and has potential to provide a standardised assessment of need in all community-nursing clients. Adequate resources are required to conduct further testing of the reliability and predictive validity of this system. RELEVANCE TO CLINICAL PRACTICE: The CCNCS can provide objective evidence of community nursing workload and thus facilitate workforce planning. PMID- 18416783 TI - Development and psychometric evaluation of workplace psychologically violent behaviours instrument. AB - AIM: To develop an instrument to determine nurses' perceptions of psychologically violent behaviours that they are exposed to in the workplace. BACKGROUND: According to Leymann, psychological terror or mobbing in work life involves hostile and unethical communication, which is directed in a systematic way towards one individual who, due to mobbing, is pushed into a helpless and defenceless position, and being held there by means of continuing mobbing activities. DESIGN: Survey. METHODS: Because nurses who work in hospitals are generally the principle victims of physical, emotional and verbal violence due to the nature of their work environment, the research sample comprised 476 hospital nurses. Data were collected via self-administered questionnaires. RESULTS: The instrument to determine the perception of workplace psychologically violent behaviours contains 33 items and four factors (individual's isolation from work, attack on professional status, attack on personality and direct attack). All items have shown statistically significant correlation (p < 0.01); the instrument's total Cronbach's alpha internal consistency coefficient was found to be 0.93. CONCLUSIONS: The findings show that the instrument's validity and reliability are within the limits of an acceptable level and that it is an instrument that will encourage more studies on this subject. RELEVANCE TO PRACTICE: Defining the psychological pressure that nurses are exposed to in the workplace and determining its negative effects on the victim of workplace psychological pressure and on the institution will make it possible to protect individuals and the institution from psychological violence with both individual and institutional practices. PMID- 18416784 TI - An exploratory study on the perceptions of healthy ageing among Chinese adults in Hong Kong. PMID- 18416785 TI - Commentary on Lee P (2007) What does partnership in care mean for children's nurses? Journal of Clinical Nursing 16, 518-526. PMID- 18416786 TI - Commentary on McSherry, R., Mudd, D., Campbell, S. (2007) Evaluating the perceived role of the nurse consultant through the lived experience of healthcare professionals. Journal of Clinical Nursing 16, 2066-2080. PMID- 18416789 TI - A best-evidence synthesis review of the administration of psychotropic pro re nata (PRN) medication in in-patient mental health settings. AB - AIMS AND OBJECTIVES: This paper aims to synthesise published literature of drug use/administration studies of pro re nata psychotropic medications in mental health wards. DESIGN: The study employed a best-evidence synthesis review design. BACKGROUND: The administration of psychotropic pro re nata medications is a frequently used clinical intervention in mental health wards. Pro re nata contributes to exposing patients to high doses of antipsychotic medication. Despite the frequent use of pro re nata, there is limited evidence of their effectiveness. METHODS: A best-evidence synthesis review. RESULTS: Six major themes emerged from the literature: (i) frequency of administration; (ii) administration during the 24-hour day; (iii) administration associated with length and stage of admission; (iv) rationales for administration; (v) medicines administered (including route of administration); and (vi) effects and side effects of the medicines administered. CONCLUSIONS: Overall findings indicate that the administration of psychotropic pro re nata varies radically and appears to be influenced by many variables. Relevance to clinical practice. Patients are most likely to receive a benzodiazepine or typical antipsychotic as pro re nata. Pro re nata is an important and under-researched clinical intervention used in mental health wards. PMID- 18416790 TI - A systematic review of professional support interventions for breastfeeding. AB - OBJECTIVES: The objectives of this systematic review were first, to describe how breastfeeding is professionally supported during pregnancy, at maternity hospitals and during the postnatal period. Secondly, to find out how effective interventions are in supporting breastfeeding. BACKGROUND: Breastfeeding is an effective way to promote the health of infants. In many countries, the rates for breastfeeding remain lower than recommended. Many studies have examined breastfeeding promotion interventions; some of them are successful and some fail. It is important to find effective combinations of support. DESIGN: Systematic review. METHODS: Search of CINAHL, Medline and Cochrane Central Register databases were conducted for data collection. The search was limited to articles published in Finnish, Swedish and English between the year 2000 and March 2006, focusing on breastfeeding and breastfeeding support interventions. Two reviewers independently analysed 36 articles in the final analysis. RESULTS: Interventions expanding from pregnancy to the intrapartum period and throughout the postnatal period were more effective than interventions concentrating on a shorter period. In addition, intervention packages using various methods of education and support from well-trained professionals are more effective than interventions concentrating on a single method. CONCLUSIONS: During pregnancy, the effective interventions were interactive, involving mothers in conversation. The Baby Friendly Hospital Initiative (BFHI) as well as practical hands off -teaching, when combined with support and encouragement, were effective approaches. Postnatally effective were home visits, telephone support and breastfeeding centres combined with peer support. Relevance to clinical practice. Professionals need breastfeeding education and support of their organisations to act as breastfeeding supporters. The BFHI -programme is effective and it would be wise to include the core components of the programme in breastfeeding promotion interventions. Mothers benefit from breastfeeding encouragement and guidance that supports their self-efficacy and feelings of being capable and empowered, and is tailored to their individual needs. PMID- 18416791 TI - A critical literature review exploring the challenges of delivering effective palliative care to older people with dementia. AB - AIM: This paper considers the challenges of delivering effective palliative care to older people with dementia and the possible strategies to overcome barriers to end-of-life care in these patients. BACKGROUND: In UK alone, approximately 100,000 people with dementia die each year and as the number of older people increases, dementia is set to become even more prevalent. Dementia is a progressive terminal illness for which there is currently no cure. Patients dying with dementia have significant health-care needs and in recent years it has been recognised that palliative care should be made available to everyone regardless of diagnosis, as this improves comfort and quality of life. Despite this, patients dying with dementia are often still not given access to palliative care services. METHOD: A review of English language literature published after 1996 to the present day relating to older people with dementia during the terminal phase of their illness. RESULTS: Twenty-nine articles met inclusion criteria for the review. Most originated from North America and UK and were mostly quantitative in nature. Four key themes were identified: difficulties associated with diagnosing the terminal phase of the illness (prognostication); issues relating to communication; medical interventions; and the appropriateness of palliative care intervention. CONCLUSIONS: This review reinforces the importance of providing appropriate palliative care to individuals suffering from end-stage dementia and identifies some of the barriers to extending such specialist palliative care provision. RELEVANCE TO PRACTICE: There is an urgent need to improve palliative care provision for older people with end-stage dementia and, in addition, more research is required on the needs of patients entering the terminal phase of dementia to assist the allocation of appropriate resources and training to ensure quality and equality in the provision of end-of-life care. PMID- 18416792 TI - A systematic review of the use of hydrocolloids in the treatment of pressure ulcers. AB - AIMS AND OBJECTIVES: The aim of this systematic literature review was to describe the current evidence in the field of pressure ulcer treatment with hydrocolloids and to give recommendations for clinical practice and further research. BACKGROUND: Pressure ulcers are a common problem in clinical practice and generate substantial expense. A wide range of dressings is available but little is known about the effect on pressure ulcer healing. METHODS: A Cochrane-based search strategy was used in four databases (PubMed, Embase, the Cochrane Central Register of Controlled Trials and the Cumulative Index to Nursing and Allied Health Literature), manuals and reference lists. Randomised controlled trials on the treatment of pressure ulcers with hydrocolloids, as defined by the British National Formulary, were systematically included and analysed. RESULTS: Twenty nine publications, dealing with 28 different studies, met the inclusion criteria and were included in the review. Hydrocolloids were most frequently used on pressure ulcers grade 2-3. Concerning the healing of the pressure ulcer, hydrocolloids are more effective than gauze dressings for the reduction of the wound dimensions. The absorption capacity, the time needed for dressing changes, the pain during dressing changes and the side-effects were significantly in favour of hydrocolloids if compared to gauze dressings. Based on the available cost-effectiveness data, hydrocolloids seemed to be less expensive compared with collagen-, saline- and povidine-soaked gauze but more expensive compared to hydrogel, polyurethane foam and collagenase. CONCLUSIONS: This review demonstrates that hydrocolloids are to be preferred to gauze dressings in the treatment of pressure ulcers. Additional research is needed to confirm these results. RELEVANCE TO CLINICAL PRACTICE: Based on the studies included in this review, hydrocolloids are frequently used in the treatment of grade 2 and 3 pressure ulcers and are more effective and less expensive than gauze dressings. Compared with alginates, polyurethane dressings, less-contact layers, topical enzymes and biosynthetic dressings, hydrocolloids are less effective. PMID- 18416793 TI - Acupressure and meridian massage: combined effects on increasing body weight in premature infants. AB - AIMS AND OBJECTIVES: The purpose of this study was to assess the body weight gain of combined acupressure and meridian massage in premature infants. BACKGROUND: Each year in Taiwan, 8-10% of newborns are delivered prematurely. Effective postnatal care is essential to ensure the healthy development of premature infants. Massage therapy has been administered to facilitate weight gain in the neonatal unit. The evidence-based findings in previous studies did not include acupressure or meridian massage. DESIGN: A double-blind clinical trial was conducted in a medical centre in central Taiwan. METHODS: Forty subjects were randomised into two groups. The 20 subjects in the experimental group were given a standard procedure of acupressure at Zhongwan, Zusanli, Yongquan, abdominal rubbing, spleen and stomach meridian massage, and kneading the points along the spine of the bladder meridian. These treatments were administered for 15 minutes per session, one hour before meals, three times daily over 10 days. The control group, also consisting of 20 subjects, underwent routine care and was observed. The infants' body weights and the volume of milk ingested were measured and recorded daily. RESULTS: The daily average weight gain of the infants in the experimental group was 32.7 g (SD = 8.1) compared with 27.3 g (SD = 7.7) in the control group. While in the first week there was no significant difference in weight gain between the two groups; in the second week, the weight gain observed in the experimental group was significantly higher than that observed in the control group. CONCLUSIONS: An experimental trial established the effects of using acupressure and meridian massage on increasing body weight in premature infants. Acupressure and meridian massage have a significant effect on weight gain in premature infants. RELEVANCE TO CLINICAL PRACTICE: Nurses could be trained in acupressure and meridian massage techniques to provide more effective level of clinical care for premature infants. PMID- 18416794 TI - Against all odds: a retrospective case-controlled study of women who experienced extraordinary breastfeeding problems. AB - AIMS: The study investigated factors empowering women to continue breastfeeding despite experiencing extraordinary difficulties. The study documented the experiences and characteristics of women who continued to breastfeed (continuing cohort) and those who weaned (non-continuing cohort) despite extraordinary difficulties. DESIGN: Retrospective case control. METHODS: The study was undertaken in south-east Queensland, Australia in 2004. Forty women (20 in each cohort) were recruited over six months. Both quantitative (breastfeeding knowledge questionnaire) and qualitative (semi-structured interviews) data were collected. This paper describes the qualitative data. RESULTS: Women from both cohorts expressed idealistic expectations about breastfeeding and experienced psychological distress due to their breastfeeding problems. Those who continued breastfeeding used coping strategies and exhibited personal qualities that assisted them to overcome the difficulties experienced. Women who continued to breastfeed were more likely to report relying on a health professional they could trust for support. This latter cohort were also more likely to report having peers with which they shared their experiences. Non-continuing women expressed feelings of guilt and inadequacy following weaning and were more likely to feel isolated. CONCLUSIONS: This study has highlighted the methods women use to deal with breastfeeding problems. It has also revealed modificable factors that can improve breastfeeding duration. RELEVANCE TO CLINICAL PRACTICE: The findings indicate that clinicians should: *Provide information which accurately reflects the breastfeeding experience; *Ensure systems are in place so that effective postnatal support for breastfeeding difficulties is available; *Consider screening to ascertain levels of psychological distress, sadness and disillusionment among breastfeeding women; *Design educational interventions with elements of cognitive skills, problem-solving and self-efficacy training to equip women with the skills to overcome any experienced difficulties. PMID- 18416795 TI - Predicting child abuse and neglect: ethical, theoretical and methodological challenges. AB - AIMS: This paper explores the ethical, theoretical and methodological implications of predictive testing in studies of child abuse, neglect and harm. BACKGROUND: The use of screening instruments to make predictions about children who may later be abused, neglected or at risk of exclusion is promoted in research and policy. The recent UK Action Plan on Social Exclusion suggests that health visitors and community midwives should be trained to use these tools in practice. METHOD: Position paper. CONCLUSIONS: The accuracy of screening instruments to identify children who will be abused or neglected has not been established. Even if the theoretical and methodological challenges of predictive instruments could be overcome, the ethical implications of their use are very difficult for nurses and midwives. RELEVANCE TO CLINICAL PRACTICE: Nurses, midwives and health visitors require high levels of awareness of risk factors in working with children and families. We have a responsibility to reach professional judgements about risks of harm for individuals. This is best performed through improving skills in working with parents and carers to identify those factors which may impede their ability to offer safe developmental care to their children. PMID- 18416796 TI - Development and psychometric testing of a Chinese version of the Fatigue Scale Children in Taiwan. AB - AIMS: The aim of this study was to develop and evaluate the psychometric properties of a Chinese version of the Fatigue Scale-Children (FS-C-C). BACKGROUND: Cancer-related fatigue is one of the most distressing and prevalent symptoms reported by paediatric oncology patients during and after their cancer treatment. A reliable and valid instrument to measure fatigue is essential to the successful clinical care of paediatric oncology patients in Taiwan who experience this troubling symptom. DESIGN AND METHODS: A total of 108 paediatric oncology Taiwanese patients who were aged 7-12 years participated in this cross-sectional, instrumentation study. The FS-C was translated into Chinese by using an established translation/back-translation method. The psychometric testing of the FS-C-C included internal consistency, content validity, construct validity, convergent validity, criterion-related validity and known-group validity. RESULTS: The FS-C-C achieved an acceptable internal consistency coefficient. Content validity was moderately high (content validity indices ranged from 83 100%). Confirmatory factor analysis supported the three-factor model as an acceptable model fit (the goodness-of-fit index and adjusted goodness-of-fit index both were greater than 0.9; normed fit index, 0.89; root-mean-square residual, 0.15). Although the results did not support the known-group hypothesis, the convergent validity, concurrent validity and predictive validity of the FS-C C were supported by significant associations with the PedsQL Multidimensional Fatigue Scale, the Anxious/Depressed subscale of the Child Behaviour Checklist and the generic and disease-specific scores on the Quality of Life for Children with Cancer instrument, respectively. CONCLUSION: Our newly developed FS-C-C is a reliable and valid instrument for the measurement of cancer-related fatigue intensity in Taiwanese children. RELEVANCE TO CLINICAL PRACTICE: The FS-C-C could provide useful information to guide clinical practice in assessing and managing cancer-related fatigue in childhood cancer patients in Taiwan. PMID- 18416797 TI - Differences in health-related quality of life between older nursing home residents without cognitive impairment and the general population of Norway. AB - AIM: To advance the understanding of health-related quality of life among older nursing home residents by assessing their health-related quality of life and comparing this with norms from the general population. METHODS: The study used a two-group cross-sectional comparative design. The samples comprised 227 nursing home residents aged 65-102 years with at least six months' residence and a representative population sample of 1137 Norwegian citizens aged 65-102 years. All nursing home residents had a Clinical Dementia Rating Scale score > or =0.5 and were capable of conversing. The respondents provided demographic information and were surveyed using the SF-36 Health Survey. We used univariate and multivariate linear models to identify possible differences in health-related quality of life between the nursing home residents and the general population, controlled for age, sex, marital status and education. RESULTS: After adjustment for age, group, sex, marital status and education, the nursing home residents scored significantly higher on bodily pain and on physical and emotional role limitation and significantly lower on the other SF-36 subscales, except social functioning, with the largest differences for physical functioning (mean nursing home 23.2 and mean general population 62.9). The general population scores on all subscales generally increased with increasing education but not among the nursing home residents. CONCLUSIONS: The mean SF-36 scale scores differed markedly between the nursing home residents and the general population, with the nursing home residents generally scoring lower. The association with background variables known to be related to health-related quality of life differed between the groups. Healthcare professionals should increase attention to health-related quality of life among nursing home residents, periodically assess health-related quality of life and consider interventions that may improve health-related quality of life in older institutionalised populations. RELEVANCE TO CLINICAL PRACTICE: This study highlights the role of nurses and other health professionals in ensuring that nursing home residents have opportunities to improve their health-related quality of life. PMID- 18416798 TI - Differences in prevalence of pressure ulcers between the Netherlands and Germany- associations between risk, prevention and occurrence of pressure ulcers in hospitals and nursing homes. AB - AIM: This study compares pressure ulcer prevalence and prevention activities in nursing homes and hospitals within two European countries. Background. Over three years stable differences have been found between the Netherlands (NL) and Germany (GER) with higher pressure ulcer rates in the NL. As previous analyses have shown, the differences cannot be entirely explained by differences in the population's vulnerability to pressure ulcers because they still remain after risk adjustment. Therefore, the differences in prevalence must be caused by other factors. The purpose of this study is to analyse if any potential differences in preventive activities can account for the varying occurrence of pressure ulcers. METHOD: In both countries, nation-wide surveys were conducted annually using the same standardised questionnaires. Trained nurses examined all consenting patients of the voluntarily participating facilities. This examination included a skin assessment of the entire body. Data regarding risk factors, prevention and details about wounds were then collected. RESULTS: In-patients of 29 German (n = 2531) and 71 Dutch (n = 10,098) nursing homes and 39 German (n = 8515) and 60 Dutch (n = 10,237) hospitals were investigated. The use of pressure-reducing devices was more common in the NL than in GER, but all other interventions were more frequently provided to German risk patients than to their Dutch counterparts. The pressure ulcer prevalence was significantly higher in the Dutch sample. After adjusting for gender, age, Braden Score and prevention, the probability of having a pressure ulcer was 8.1 times higher for Dutch nursing home residents than for German residents. CONCLUSION: Some of the variance in pressure ulcer prevalence between the two countries can be explained by varying pressure ulcer prevention. However, some remarkable differences still remain unexplained. RELEVANCE TO CLINICAL PRACTICE: The extent of pressure ulcer prevention, especially repositioning and nutrition intervention provided to patients at risk, is not in accordance with international guidelines. PMID- 18416799 TI - Commentary on Schofield I, Kerr S & Tolson D (2007) 'An exploration of the smoking-related health beliefs of older people with chronic obstructive pulmonary disease'. Journal of Clinical Nursing 16, 1726-1735. PMID- 18416800 TI - Commentary on Mamhidir A-G, Karlsson I, Norberg A, Kihlgren M (2007) Weight increase in patients with dementia, and alteration in meal routines and meal environment after integrity promoting care. Journal of Clinical Nursing 16, 987 996. PMID- 18416801 TI - Commentary on Mantzoukas S & Watkinson S (2007) Advanced nursing practice: reviewing the international literature and developing the generic features. Journal of Clinical Nursing 16, 28-37. PMID- 18416802 TI - Commentary on Donnelly F & Wiechula R (2006) The lived experience of a tracheostomy tube change: a phenomenological study. Journal of Clinical Nursing, 15, 1115-1122. PMID- 18416803 TI - Commentary on Humphreys A, Johnson S, Richardson J, Stenhouse E, Watkins M (2007) Evaluating the effectiveness of nurse, midwife/allied health professional consultants: a systematic review and meta-synthesis Journal of Clinical Nursing 16, 1792-1808. PMID- 18416804 TI - Commentary on Jolley J (2007) Choose your doctorate. Journal of Clinical Nursing 16, 225-233. PMID- 18416806 TI - Commentary on Josefsson K, Sonde L & Robins Wahlin T-B (2007) Violence in municipal care of older people in Sweden as perceived by registered nurses. Journal of Clinical Nursing 16, 900-910. PMID- 18416808 TI - Commentary on Eley R, Hegney D, Buikstra E, Fallon T, Plank A & Parker V (2007) Aged care nursing in Queensland--the nurses' view. Journal of Clinical Nursing, 16, 860-872. PMID- 18416810 TI - How many human genes can be defined as housekeeping with current expression data? AB - BACKGROUND: Housekeeping (HK) genes are ubiquitously expressed in all tissue/cell types and constitute a basal transcriptome for the maintenance of basic cellular functions. Partitioning transcriptomes into HK and tissue-specific (TS) genes relatively is fundamental for studying gene expression and cellular differentiation. Although many studies have aimed at large-scale and thorough categorization of human HK genes, a meaningful consensus has yet to be reached. RESULTS: We collected two latest gene expression datasets (both EST and microarray data) from public databases and analyzed the gene expression profiles in 18 human tissues that have been well-documented by both two data types. Benchmarked by a manually-curated HK gene collection (HK408), we demonstrated that present data from EST sampling was far from saturated, and the inadequacy has limited the gene detectability and our understanding of TS expressions. Due to a likely over-stringent threshold, microarray data showed higher false negative rate compared with EST data, leading to a significant underestimation of HK genes. Based on EST data, we found that 40.0% of the currently annotated human genes were universally expressed in at least 16 of 18 tissues, as compared to only 5.1% specifically expressed in a single tissue. Our current EST-based estimate on human HK genes ranged from 3,140 to 6,909 in number, a ten-fold increase in comparison with previous microarray-based estimates. CONCLUSION: We concluded that a significant fraction of human genes, at least in the currently annotated data depositories, was broadly expressed. Our understanding of tissue specific expression was still preliminary and required much more large-scale and high-quality transcriptomic data in future studies. The new HK gene list categorized in this study will be useful for genome-wide analyses on structural and functional features of HK genes. PMID- 18416811 TI - Transcriptome architecture across tissues in the pig. AB - BACKGROUND: Artificial selection has resulted in animal breeds with extreme phenotypes. As an organism is made up of many different tissues and organs, each with its own genetic programme, it is pertinent to ask: How relevant is tissue in terms of total transcriptome variability? Which are the genes most distinctly expressed between tissues? Does breed or sex equally affect the transcriptome across tissues? RESULTS: In order to gain insight on these issues, we conducted microarray expression profiling of 16 different tissues from four animals of two extreme pig breeds, Large White and Iberian, two males and two females. Mixed model analysis and neighbor - joining trees showed that tissues with similar developmental origin clustered closer than those with different embryonic origins. Often a sound biological interpretation was possible for overrepresented gene ontology categories within differentially expressed genes between groups of tissues. For instance, an excess of nervous system or muscle development genes were found among tissues of ectoderm or mesoderm origins, respectively. Tissue accounted for ~11 times more variability than sex or breed. Nevertheless, we were able to confidently identify genes with differential expression across tissues between breeds (33 genes) and between sexes (19 genes). The genes primarily affected by sex were overall different than those affected by breed or tissue. Interaction with tissue can be important for differentially expressed genes between breeds but not so much for genes whose expression differ between sexes. CONCLUSION: Embryonic development leaves an enduring footprint on the transcriptome. The interaction in gene x tissue for differentially expressed genes between breeds suggests that animal breeding has targeted differentially each tissue's transcriptome. PMID- 18416812 TI - Conducting a meta-ethnography of qualitative literature: lessons learnt. AB - BACKGROUND: Qualitative synthesis has become more commonplace in recent years. Meta-ethnography is one of several methods for synthesising qualitative research and is being used increasingly within health care research. However, many aspects of the steps in the process remain ill-defined. DISCUSSION: We utilized the seven stages of the synthesis process to synthesise qualitative research on adherence to tuberculosis treatment. In this paper we discuss the methodological and practical challenges faced; of particular note are the methods used in our synthesis, the additional steps that we found useful in clarifying the process, and the key methodological challenges encountered in implementing the meta ethnographic approach. The challenges included shaping an appropriate question for the synthesis; identifying relevant studies; assessing the quality of the studies; and synthesising findings across a very large number of primary studies from different contexts and research traditions. We offer suggestions that may assist in undertaking meta-ethnographies in the future. SUMMARY: Meta-ethnography is a useful method for synthesising qualitative research and for developing models that interpret findings across multiple studies. Despite its growing use in health research, further research is needed to address the wide range of methodological and epistemological questions raised by the approach. PMID- 18416813 TI - Non-random retention of protein-coding overlapping genes in Metazoa. AB - BACKGROUND: Although the overlap of transcriptional units occurs frequently in eukaryotic genomes, its evolutionary and biological significance remains largely unclear. Here we report a comparative analysis of overlaps between genes coding for well-annotated proteins in five metazoan genomes (human, mouse, zebrafish, fruit fly and worm). RESULTS: For all analyzed species the observed number of overlapping genes is always lower than expected assuming functional neutrality, suggesting that gene overlap is negatively selected. The comparison to the random distribution also shows that retained overlaps do not exhibit random features: antiparallel overlaps are significantly enriched, while overlaps lying on the same strand and those involving coding sequences are highly underrepresented. We confirm that overlap is mostly species-specific and provide evidence that it frequently originates through the acquisition of terminal, non-coding exons. Finally, we show that overlapping genes tend to be significantly co-expressed in a breast cancer cDNA library obtained by 454 deep sequencing, and that different overlap types display different patterns of reciprocal expression. CONCLUSION: Our data suggest that overlap between protein-coding genes is selected against in Metazoa. However, when retained it may be used as a species-specific mechanism for the reciprocal regulation of neighboring genes. The tendency of overlaps to involve non-coding regions of the genes leads to the speculation that the advantages achieved by an overlapping arrangement may be optimized by evolving regulatory non-coding transcripts. PMID- 18416814 TI - anNET: a tool for network-embedded thermodynamic analysis of quantitative metabolome data. AB - BACKGROUND: Compared to other omics techniques, quantitative metabolomics is still at its infancy. Complex sample preparation and analytical procedures render exact quantification extremely difficult. Furthermore, not only the actual measurement but also the subsequent interpretation of quantitative metabolome data to obtain mechanistic insights is still lacking behind the current expectations. Recently, the method of network-embedded thermodynamic (NET) analysis was introduced to address some of these open issues. Building upon principles of thermodynamics, this method allows for a quality check of measured metabolite concentrations and enables to spot metabolic reactions where active regulation potentially controls metabolic flux. So far, however, widespread application of NET analysis in metabolomics labs was hindered by the absence of suitable software. RESULTS: We have developed in Matlab a generalized software called 'anNET' that affords a user-friendly implementation of the NET analysis algorithm. anNET supports the analysis of any metabolic network for which a stoichiometric model can be compiled. The model size can span from a single reaction to a complete genome-wide network reconstruction including compartments. anNET can (i) test quantitative data sets for thermodynamic consistency, (ii) predict metabolite concentrations beyond the actually measured data, (iii) identify putative sites of active regulation in the metabolic reaction network, and (iv) help in localizing errors in data sets that were found to be thermodynamically infeasible. We demonstrate the application of anNET with three published Escherichia coli metabolome data sets. CONCLUSION: Our user-friendly and generalized implementation of the NET analysis method in the software anNET allows users to rapidly integrate quantitative metabolome data obtained from virtually any organism. We envision that use of anNET in labs working on quantitative metabolomics will provide the systems biology and metabolic engineering communities with a mean to proof the quality of metabolome data sets and with all further benefits of the NET analysis approach. PMID- 18416815 TI - Marked variation in predicted and observed variability of tandem repeat loci across the human genome. AB - BACKGROUND: Tandem repeat (TR) variants in the human genome play key roles in a number of diseases. However, current models predicting variability are based on limited training sets. We conducted a systematic analysis of TRs of unit lengths 2-12 nucleotides in Whole Genome Shotgun (WGS) sequences to define the extent of variation of 209,214 unique repeat loci throughout the genome. RESULTS: We applied a multivariate statistical model to predict TR variability. Predicted heterozygosity correlated with heterozygosity in the CEPH polymorphism database (correlation rho = 0.29, p < 0.0005) better than the correlation between the CEPH and WGS data (rho = 0.17), presumably because the model smoothes noise from small sample sizes. A multivariate logistic model of 8 parameters accounted for 36% of the variation in the WGS data. Validation studies of 70 experimentally investigated TRs revealed high concordance with the model's predictions (p < 0.0001). CONCLUSION: Variability among 2-12-mer TRs in the genome can be modeled by a few parameters, which do not markedly differ according to unit length, consistent with a common mechanism for the generation of variability among such TRs. Analysis of the distributions of observed and predicted variants across the genome showed a general concordance, indicating that the repeat variation dataset does not exhibit strong regional ascertainment biases. This revealed a deficit of variant repeats in chromosomes 19 and Y - likely to reflect a reduction in 2-mer repeats in the former and a reduced level of recombination in the latter - and excesses in chromosomes 6, 13, 20 and 21. PMID- 18416816 TI - Gene transfer of Hodgkin cell lines via multivalent anti-CD30 scFv displaying bacteriophage. AB - BACKGROUND: The display of binding ligands, such as recombinant antibody fragments, on the surface of filamentous phage makes it possible to specifically attach these phage particles to target cells. After uptake of the phage, their internal single-stranded DNA is processed by the host cell, which allows transient expression of an encoded eukaryotic gene cassette. This opens the possibility to use bacteriophage as vectors for targeted gene therapy, although the transduction efficiency is very low. RESULTS: Here we demonstrate the display of an anti-CD30 single chain variable fragment fused to the major coat protein pVIII on the surface of bacteriophage. These phage particles showed an improved binding and transduction efficiency of CD30 positive Hodgkin-lymphoma cells, compared to bacteriophage with the anti-CD30 single chain variable fragment fused to the minor coat protein pIII. CONCLUSION: We can conclude from the results that the postulated multivalency of the anti-CD30-pVIII displaying bacteriophage combined with disseminated display of the anti-CD30 scFv on the whole particle surface is responsible for the improved gene transfer rate. These results mark an important step towards the use of phage particles as a cheap and safe gene transfer vehicle for the gene delivery of the desired target cells via their specific surface receptors. PMID- 18416817 TI - ATBF1 and NQO1 as candidate targets for allelic loss at chromosome arm 16q in breast cancer: absence of somatic ATBF1 mutations and no role for the C609T NQO1 polymorphism. AB - BACKGROUND: Loss of heterozygosity (LOH) at chromosome arm 16q is frequently observed in human breast cancer, suggesting that one or more target tumor suppressor genes (TSGs) are located there. However, detailed mapping of the smallest region of LOH has not yet resulted in the identification of a TSG at 16q. Therefore, the present study attempted to identify TSGs using an approach based on mRNA expression. METHODS: A cDNA microarray for the 16q region was constructed and analyzed using RNA samples from 39 breast tumors with known LOH status at 16q. RESULTS: Five genes were identified to show lower expression in tumors with LOH at 16q compared to tumors without LOH. The genes for NAD(P)H dehydrogenase quinone (NQO1) and AT-binding transcription factor 1 (ATBF1) were further investigated given their functions as potential TSGs. NQO1 has been implicated in carcinogenesis due to its role in quinone detoxification and in stabilization of p53. One inactive polymorphic variant of NQO1 encodes a product showing reduced enzymatic activity. However, we did not find preferential targeting of the active NQO1 allele in tumors with LOH at 16q. Immunohistochemical analysis of 354 invasive breast tumors revealed that NQO1 protein expression in a subset of breast tumors is higher than in normal epithelium, which contradicts its proposed role as a tumor suppressor gene.ATBF1 has been suggested as a target for LOH at 16q in prostate cancer. We analyzed the entire coding sequence in 48 breast tumors, but did not identify somatic sequence changes. We did find several in-frame insertions and deletions, two variants of which were reported to be somatic pathogenic mutations in prostate cancer. Here, we show that these variants are also present in the germline in 2.5% of 550 breast cancer patients and 2.9% of 175 healthy controls. This indicates that the frequency of these variants is not increased in breast cancer patients. Moreover, there is no preferential LOH of the wildtype allele in breast tumors. CONCLUSION: Two likely candidate TSGs at 16q in breast cancer, NQO1 and ATBF1, were identified here as showing reduced expression in tumors with 16q LOH, but further analysis indicated that they are not target genes of LOH. Furthermore, our results call into question the validity of the previously reported pathogenic variants of the ATBF1 gene. PMID- 18416818 TI - The educational background and qualifications of UK medical students from ethnic minorities. AB - BACKGROUND: UK medical students and doctors from ethnic minorities underperform in undergraduate and postgraduate examinations. Although it is assumed that white (W) and non-white (NW) students enter medical school with similar qualifications, neither the qualifications of NW students, nor their educational background have been looked at in detail. This study uses two large-scale databases to examine the educational attainment of W and NW students. METHODS: Attainment at GCSE and A level, and selection for medical school in relation to ethnicity, were analysed in two separate databases. The 10th cohort of the Youth Cohort Study provided data on 13,698 students taking GCSEs in 1999 in England and Wales, and their subsequent progression to A level. UCAS provided data for 1,484,650 applicants applying for admission to UK universities and colleges in 2003, 2004 and 2005, of whom 52,557 applied to medical school, and 23,443 were accepted. RESULTS: NW students achieve lower grades at GCSE overall, although achievement at the highest grades was similar to that of W students. NW students have higher educational aspirations, being more likely to go on to take A levels, especially in science and particularly chemistry, despite relatively lower achievement at GCSE. As a result, NW students perform less well at A level than W students, and hence NW students applying to university also have lower A-level grades than W students, both generally, and for medical school applicants. NW medical school entrants have lower A level grades than W entrants, with an effect size of about 0.10. CONCLUSION: The effect size for the difference between white and non-white medical school entrants is about B0.10, which would mean that for a typical medical school examination there might be about 5 NW failures for each 4 W failures. However, this effect can only explain a portion of the overall effect size found in undergraduate and postgraduate examinations of about -0.32. PMID- 18416819 TI - Are the determinants of vertebral endplate changes and severe disc degeneration in the lumbar spine the same? A magnetic resonance imaging study in middle-aged male workers. AB - BACKGROUND: Modic changes are bone marrow lesions visible in magnetic resonance imaging (MRI), and they are assumed to be associated with symptomatic intervertebral disc disease, especially changes located at L5-S1. Only limited information exists about the determinants of Modic changes. The objective of this study was to evaluate the determinants of vertebral endplate (Modic) changes, and whether they are similar for Modic changes and severe disc degeneration focusing on L5-S1 level. METHODS: 228 middle-aged male workers (159 train engineers and 69 sedentary factory workers) from northern Finland underwent sagittal T1- and T2 weighted MRI. Modic changes and disc degeneration were analyzed from the scans. The participants responded to a questionnaire including items of occupational history and lifestyle factors. Logistic regression analysis was used to evaluate the associations between selected determinants (age, lifetime exercise, weight related factors, fat percentage, smoking, alcohol use, lifetime whole-body vibration) and Modic type I and II changes, and severe disc degeneration (= grade V on Pfirrmann's classification). RESULTS: The prevalences of the Modic changes and severe disc degeneration were similar in the occupational groups. Age was significantly associated with all degenerative changes. In the age-adjusted analyses, only weight-related determinants (BMI, waist circumference) were associated with type II changes. Exposure to whole-body vibration, besides age, was the only significant determinant for severe disc degeneration. In the multivariate model, BMI was associated with type II changes at L5-S1 (OR 2.75 per one SD = 3 unit increment in BMI), and vibration exposure with severe disc degeneration at L5-S1 (OR 1.08 per one SD = 11-year increment in vibration exposure). CONCLUSION: Besides age, weight-related factors seem important in the pathogenesis of Modic changes, whereas whole-body vibration was the only significant determinant of severe disc degeneration. PMID- 18416821 TI - Association between amebic liver abscess and human immunodeficiency virus infection in Taiwanese subjects. AB - PURPOSE: Invasive amebiasis is an emerging parasitic disorder in Taiwan, especially in patients diagnosed with human immunodeficiency virus (HIV) infection. Thirty-three Taiwanese subjects with amebic liver abscess (ALA) were examined and a possible correlation between ALA and HIV infection was investigated. RESULTS: Among ALA patients, the proportion of HIV-positive individuals increased during the study period. ALA was the first major clinical presentation in 54% of HIV patients with ALA. Overall, 58% (14/24) of HIV infected patients had a CD4+ count > 200 cells/muL and 82.1% (23/28) had no concurrent opportunistic infection or other evidence of HIV infection. There was no marked difference in clinical characteristics between HIV-positive and HIV negative ALA patients except the level of leukocytosis. CONCLUSION: While the clinical characteristics described herein cannot be used to determine whether ALA patients have HIV infection, routine HIV testing is recommended in patients with ALA, even in the absence of HIV symptoms. PMID- 18416820 TI - Assessment of CcpA-mediated catabolite control of gene expression in Bacillus cereus ATCC 14579. AB - BACKGROUND: The catabolite control protein CcpA is a transcriptional regulator conserved in many Gram-positives, controlling the efficiency of glucose metabolism. Here we studied the role of Bacillus cereus ATCC 14579 CcpA in regulation of metabolic pathways and expression of enterotoxin genes by comparative transcriptome analysis of the wild-type and a ccpA-deletion strain. RESULTS: Comparative analysis revealed the growth performance and glucose consumption rates to be lower in the B. cereus ATCC 14579 ccpA deletion strain than in the wild-type. In exponentially grown cells, the expression of glycolytic genes, including a non-phosphorylating glyceraldehyde-3-phosphate dehydrogenase that mediates conversion of D-glyceraldehyde 3-phosphate to 3-phospho-D-glycerate in one single step, was down-regulated and expression of gluconeogenic genes and genes encoding the citric acid cycle was up-regulated in the B. cereus ccpA deletion strain. Furthermore, putative CRE-sites, that act as binding sites for CcpA, were identified to be present for these genes. These results indicate CcpA to be involved in the regulation of glucose metabolism, thereby optimizing the efficiency of glucose catabolism. Other genes of which the expression was affected by ccpA deletion and for which putative CRE-sites could be identified, included genes with an annotated function in the catabolism of ribose, histidine and possibly fucose/arabinose and aspartate. Notably, expression of the operons encoding non-hemolytic enterotoxin (Nhe) and hemolytic enterotoxin (Hbl) was affected by ccpA deletion, and putative CRE-sites were identified, which suggests catabolite repression of the enterotoxin operons to be CcpA-dependent. CONCLUSION: The catabolite control protein CcpA in B. cereus ATCC 14579 is involved in optimizing the catabolism of glucose with concomitant repression of gluconeogenesis and alternative metabolic pathways. Furthermore, the results point to metabolic control of enterotoxin gene expression and suggest that CcpA mediated glucose sensing provides an additional mode of control in moderating the expression of the nhe and hbl operons in B. cereus ATCC 14579. PMID- 18416822 TI - Characterisation of the cannabinoid receptor system in synovial tissue and fluid in patients with osteoarthritis and rheumatoid arthritis. AB - INTRODUCTION: Cannabis-based medicines have a number of therapeutic indications, including anti-inflammatory and analgesic effects. The endocannabinoid receptor system, including the cannabinoid receptor 1 (CB1) and receptor 2 (CB2) and the endocannabinoids, are implicated in a wide range of physiological and pathophysiological processes. Pre-clinical and clinical studies have demonstrated that cannabis-based drugs have therapeutic potential in inflammatory diseases, including rheumatoid arthritis (RA) and multiple sclerosis. The aim of this study was to determine whether the key elements of the endocannabinoid signalling system, which produces immunosuppression and analgesia, are expressed in the synovia of patients with osteoarthritis (OA) or RA. METHODS: Thirty-two OA and 13 RA patients undergoing total knee arthroplasty were included in this study. Clinical staging was conducted from x-rays scored according to Kellgren-Lawrence and Larsen scales, and synovitis of synovial biopsies was graded. Endocannabinoid levels were quantified in synovial fluid by liquid chromatography-mass spectrometry. The expression of CB1 and CB2 protein and RNA in synovial biopsies was investigated. Functional activity of these receptors was determined with mitogen-activated protein kinase assays. To assess the impact of OA and RA on this receptor system, levels of endocannabinoids in the synovial fluid of patients and non-inflamed healthy volunteers were compared. The activity of fatty acid amide hydrolase (FAAH), the predominant catabolic endocannabinoid enzyme, was measured in synovium. RESULTS: CB1 and CB2 protein and RNA were present in the synovia of OA and RA patients. Cannabinoid receptor stimulation of fibroblast like cells from OA and RA patients produced a time-dependent phosphorylation of extracellular signal-regulated kinase (ERK)-1 and ERK-2 which was significantly blocked by the CB1 antagonist SR141716A. The endocannabinoids anandamide (AEA) and 2-arachidonyl glycerol (2-AG) were identified in the synovial fluid of OA and RA patients. However, neither AEA nor 2-AG was detected in synovial fluid from normal volunteers. FAAH was active in the synovia of OA and RA patients and was sensitive to inhibition by URB597 (3'-(aminocarbonyl) [1,1'-biphenyl]-3-yl) cyclohexylcarbamate). CONCLUSION: Our data predict that the cannabinoid receptor system present in the synovium may be an important therapeutic target for the treatment of pain and inflammation associated with OA and RA. PMID- 18416823 TI - Raisins and additional walking have distinct effects on plasma lipids and inflammatory cytokines. AB - BACKGROUND: Raisins are a significant source of dietary fiber and polyphenols, which may reduce cardiovascular disease (CVD) risk by affecting lipoprotein metabolism and inflammation. Walking represents a low intensity exercise intervention that may also reduce CVD risk. The purpose of this study was to determine the effects of consuming raisins, increasing steps walked, or a combination of these interventions on blood pressure, plasma lipids, glucose, insulin and inflammatory cytokines. RESULTS: Thirty-four men and postmenopausal women were matched for weight and gender and randomly assigned to consume 1 cup raisins/d (RAISIN), increase the amount of steps walked/d (WALK) or a combination of both interventions (RAISINS + WALK). The subjects completed a 2 wk run-in period, followed by a 6 wk intervention. Systolic blood pressure was reduced for all subjects (P = 0.008). Plasma total cholesterol was decreased by 9.4% for all subjects (P < 0.005), which was explained by a 13.7% reduction in plasma LDL cholesterol (LDL-C) (P < 0.001). Plasma triglycerides (TG) concentrations were decreased by 19.5% for WALK (P < 0.05 for group effect). Plasma TNF-alpha was decreased from 3.5 ng/L to 2.1 ng/L for RAISIN (P < 0.025 for time and group x time effect). All subjects had a reduction in plasma sICAM-1 (P < 0.01). CONCLUSION: This research shows that simple lifestyle modifications such as adding raisins to the diet or increasing steps walked have distinct beneficial effects on CVD risk. PMID- 18416824 TI - Release of glutamate and CGRP from trigeminal ganglion neurons: Role of calcium channels and 5-HT1 receptor signaling. AB - BACKGROUND: The aberrant release of the neurotransmitters, glutamate and calcitonin-gene related peptide (CGRP), from trigeminal neurons has been implicated in migraine. The voltage-gated P/Q-type calcium channel has a critical role in controlling neurotransmitter release and has been linked to Familial Hemiplegic Migraine. Therefore, we examined the importance of voltage-dependent calcium channels in controlling release of glutamate and CGRP from trigeminal ganglion neurons isolated from male and female rats and grown in culture. Serotonergic pathways are likely involved in migraine, as triptans, a class of 5 HT1 receptor agonists, are effective in the treatment of migraine and their effectiveness may be due to inhibiting neurotransmitter release from trigeminal neurons. We also studied the effect of serotonin receptor activation on release of glutamate and CGRP from trigeminal neurons grown in culture. RESULTS: P/Q-, N- and L-type channels each mediate a significant fraction of potassium-stimulated release of glutamate and CGRP. We determined that 5-HT significantly inhibits potassium-stimulated release of both glutamate and CGRP. Serotonergic inhibition of both CGRP and glutamate release can be blocked by pertussis toxin and NAS-181, a 5-HT1B/1D antagonist. Stimulated release of CGRP is unaffected by Y-25130, a 5 HT3 antagonist and SB 200646, a 5-HT2B/2C antagonist. CONCLUSION: These data suggest that release of both glutamate and CGRP from trigeminal neurons is controlled by calcium channels and modulated by 5-HT signaling in a pertussis toxin dependent manner and probably via 5-HT1 receptor signaling. This is the first characterization of glutamate release from trigeminal neurons grown in culture. PMID- 18416825 TI - Structural insights on the pamoic acid and the 8 kDa domain of DNA polymerase beta complex: towards the design of higher-affinity inhibitors. AB - BACKGROUND: DNA polymerase beta (pol beta), the error-prone DNA polymerase of single-stranded DNA break repair as well as base excision repair pathways, is overexpressed in several tumors and takes part in chemotherapeutic agent resistance, like that of cisplatin, through translesion synthesis. For this reason pol beta has become a therapeutic target. Several inhibitors have been identified, but none of them presents a sufficient affinity and specificity to become a drug. The fragment-based inhibitor design allows an important improvement in affinity of small molecules. The initial and critical step for setting up the fragment-based strategy consists in the identification and structural characterization of the first fragment bound to the target. RESULTS: We have performed docking studies of pamoic acid, a 9 micromolar pol beta inhibitor, and found that it binds in a single pocket at the surface of the 8 kDa domain of pol beta. However, docking studies provided five possible conformations for pamoic acid in this site. NMR experiments were performed on the complex to select a single conformation among the five retained. Chemical Shift Mapping data confirmed pamoic acid binding site found by docking while NOESY and saturation transfer experiments provided distances between pairs of protons from the pamoic acid and those of the 8 kDa domain that allowed the identification of the correct conformation. CONCLUSION: Combining NMR experiments on the complex with docking results allowed us to build a three-dimensional structural model. This model serves as the starting point for further structural studies aimed at improving the affinity of pamoic acid for binding to DNA polymerase beta. PMID- 18416826 TI - Explorative data analysis of MCL reveals gene expression networks implicated in survival and prognosis supported by explorative CGH analysis. AB - BACKGROUND: Mantle cell lymphoma (MCL) is an incurable B cell lymphoma and accounts for 6% of all non-Hodgkin's lymphomas. On the genetic level, MCL is characterized by the hallmark translocation t(11;14) that is present in most cases with few exceptions. Both gene expression and comparative genomic hybridization (CGH) data vary considerably between patients with implications for their prognosis. METHODS: We compare patients over and below the median of survival. Exploratory principal component analysis of gene expression data showed that the second principal component correlates well with patient survival. Explorative analysis of CGH data shows the same correlation. RESULTS: On chromosome 7 and 9 specific genes and bands are delineated which improve prognosis prediction independent of the previously described proliferation signature. We identify a compact survival predictor of seven genes for MCL patients. After extensive re-annotation using GEPAT, we established protein networks correlating with prognosis. Well known genes (CDC2, CCND1) and further proliferation markers (WEE1, CDC25, aurora kinases, BUB1, PCNA, E2F1) form a tight interaction network, but also non-proliferative genes (SOCS1, TUBA1B CEBPB) are shown to be associated with prognosis. Furthermore we show that aggressive MCL implicates a gene network shift to higher expressed genes in late cell cycle states and refine the set of non-proliferative genes implicated with bad prognosis in MCL. CONCLUSION: The results from explorative data analysis of gene expression and CGH data are complementary to each other. Including further tests such as Wilcoxon rank test we point both to proliferative and non-proliferative gene networks implicated in inferior prognosis of MCL and identify suitable markers both in gene expression and CGH data. PMID- 18416827 TI - A statistical model for the identification of genes governing the incidence of cancer with age. AB - The cancer incidence increases with age. This epidemiological pattern of cancer incidence can be attributed to molecular and cellular processes of individual subjects. Also, the incidence of cancer with ages can be controlled by genes. Here we present a dynamic statistical model for explaining the epidemiological pattern of cancer incidence based on individual genes that regulate cancer formation and progression. We incorporate the mathematical equations of age specific cancer incidence into a framework for functional mapping aimed at identifying quantitative trait loci (QTLs) for dynamic changes of a complex trait. The mathematical parameters that specify differences in the curve of cancer incidence among QTL genotypes are estimated within the context of maximum likelihood. The model provides testable quantitative hypotheses about the initiation and duration of genetic expression for QTLs involved in cancer progression. Computer simulation was used to examine the statistical behavior of the model. The model can be used as a tool for explaining the epidemiological pattern of cancer incidence. PMID- 18416828 TI - A computational model for sex-specific genetic architecture of complex traits in humans: implications for mapping pain sensitivity. AB - Understanding differences in the genetic architecture of complex traits between the two sexes has significant implications for evolutionary studies and clinical diagnosis. However, our knowledge about sex-specific genetic architecture is limited largely because of a lack of analytical models that can detect and quantify the effects of sex on the complexity of quantitative genetic variation. Here, we derived a statistical model for mapping DNA sequence variants that contribute to sex-specific differences in allele frequencies, linkage disequilibria, and additive and dominance genetic effects due to haplotype diversity. This model allows a genome-wide search for functional haplotypes and the estimation and test of haplotype by sex interactions and sex-specific heritability. The model, validated by simulation studies, was used to detect sex specific functional haplotypes that encode a pain sensitivity trait in humans. The model could have important implications for mapping complex trait genes and studying the detailed genetic architecture of sex-specific differences. PMID- 18416829 TI - ParaKMeans: Implementation of a parallelized K-means algorithm suitable for general laboratory use. AB - BACKGROUND: During the last decade, the use of microarrays to assess the transcriptome of many biological systems has generated an enormous amount of data. A common technique used to organize and analyze microarray data is to perform cluster analysis. While many clustering algorithms have been developed, they all suffer a significant decrease in computational performance as the size of the dataset being analyzed becomes very large. For example, clustering 10000 genes from an experiment containing 200 microarrays can be quite time consuming and challenging on a desktop PC. One solution to the scalability problem of clustering algorithms is to distribute or parallelize the algorithm across multiple computers. RESULTS: The software described in this paper is a high performance multithreaded application that implements a parallelized version of the K-means Clustering algorithm. Most parallel processing applications are not accessible to the general public and require specialized software libraries (e.g. MPI) and specialized hardware configurations. The parallel nature of the application comes from the use of a web service to perform the distance calculations and cluster assignments. Here we show our parallel implementation provides significant performance gains over a wide range of datasets using as little as seven nodes. The software was written in C# and was designed in a modular fashion to provide both deployment flexibility as well as flexibility in the user interface. CONCLUSION: ParaKMeans was designed to provide the general scientific community with an easy and manageable client-server application that can be installed on a wide variety of Windows operating systems. PMID- 18416830 TI - The Retinoic Acid Receptor-alpha mediates human T-cell activation and Th2 cytokine and chemokine production. AB - BACKGROUND: We have recently demonstrated that all-trans-retinoic acid (ATRA) and 9-cis-retinoic acid (9-cis RA) promote IL-4, IL-5 and IL-13 synthesis, while decreasing IFN-gamma and TNF-alpha expression by activated human T cells and reduces the synthesis of IL-12p70 from accessory cells. Here, we have demonstrated that the observed effects using ATRA and 9-cis RA are shared with the clinically useful RAR ligand, 13-cis retinoic acid (13-cis RA), and the retinoic acid receptor-alpha (RAR-alpha)-selective agonist, AM580 but not with the RAR-beta/gamma ligand, 4-hydroxyphenylretinamide (4-HPR). RESULTS: The increase in type 2 cytokine production by these retinoids correlated with the expression of the T cell activation markers, CD69 and CD38. The RAR-alpha selective agonist, AM580 recapitulated all of the T cell activation and type 2 cytokine-inducing effects of ATRA and 9-cis-RA, while the RAR-alpha-selective antagonist, RO 41-5253, inhibited these effects. CONCLUSION: These results strongly support a role for RAR-alpha engagement in the regulation of genes and proteins involved with human T cell activation and type 2 cytokine production. PMID- 18416831 TI - Two-dimensional electrophoretic comparison of metastatic and non-metastatic human breast tumors using in vitro cultured epithelial cells derived from the cancer tissues. AB - BACKGROUND: Breast carcinomas represent a heterogeneous group of tumors diverse in behavior, outcome, and response to therapy. Identification of proteins resembling the tumor biology can improve the diagnosis, prediction, treatment selection, and targeting of therapy. Since the beginning of the post-genomic era, the focus of molecular biology gradually moved from genomes to proteins and proteomes and to their functionality. Proteomics can potentially capture dynamic changes in protein expression integrating both genetic and epigenetic influences. METHODS: We prepared primary cultures of epithelial cells from 23 breast cancer tissue samples and performed comparative proteomic analysis. Seven patients developed distant metastases within three-year follow-up. These samples were included into a metastase-positive group, the others formed a metastase-negative group. Two-dimensional electrophoretical (2-DE) gels in pH range 4-7 were prepared. Spot densities in 2-DE protein maps were subjected to statistical analyses (R/maanova package) and data-mining analysis (GUHA). For identification of proteins in selected spots, liquid chromatography-tandem mass spectrometry (LC MS/MS) was employed. RESULTS: Three protein spots were significantly altered between the metastatic and non-metastatic groups. The correlations were proven at the 0.05 significance level. Nucleophosmin was increased in the group with metastases. The levels of 2,3-trans-enoyl-CoA isomerase and glutathione peroxidase 1 were decreased. CONCLUSION: We have performed an extensive proteomic study of mammary epithelial cells from breast cancer patients. We have found differentially expressed proteins between the samples from metastase-positive and metastase-negative patient groups. PMID- 18416832 TI - Changes in viral load and HBsAg and HBeAg status with age in HBV chronic carriers in The Gambia. AB - BACKGROUND: Little is known about changes in hepatitis B viral load (HBV DNA) in relation to age in Africa. The aim of this study is to determine the natural course of HBV chronic infection, particularly in relation to sequential changes in serum HBV DNA levels and hepatitis B surface (HBsAg) antigen/hepatitis e antigen (HBeAg) status by age. METHODS: The study was conducted on 190 HBV chronic carriers, aged 1-19 years who were followed for 19 years. 160, 99 and 123 were traced at 5, 9 and 19 years later. All available samples were tested for HBsAg and HBeAg, whilst 170, 61, 63 and 81 were tested for HBV DNA at the baseline, and at 5, 9 and 19 years following recruitment. RESULTS: In general HBeAg which correlated with high levels of HBV DNA was lost at a much faster rate than HBsAg. 86% of the carriers who were recruited at the age of 1-4 yrs lost HBeAg by the age of 19 years compared to 30% who lost HBsAg. HBeAg negative carriers had serum HBV DNA levels of < 105 copies per mL, HBV DNA positivity declined from 100% in 1-4 yrs old carriers at recruitment to 62.5%,60% and 88% at 5, 9 and 19 years respectively following recruitment. CONCLUSION: After 19 years of follow up, the majority of HBV surface antigen carriers had lost HBeAg positivity and had low levels of viral replication. However small proportions (10 20%) retained HBeAg and continue to have high levels of viral replication. PMID- 18416833 TI - Directional and balancing selection in human beta-defensins. AB - BACKGROUND: In primates, infection is an important force driving gene evolution, and this is reflected in the importance of infectious disease in human morbidity today. The beta-defensins are key components of the innate immune system, with antimicrobial and cell signalling roles, but also reproductive functions. Here we examine evolution of beta-defensins in catarrhine primates and variation within different human populations. RESULTS: We show that five beta-defensin genes that do not show copy number variation in humans show evidence of positive selection in catarrhine primates, and identify specific codons that have been under selective pressure. Direct haplotyping of DEFB127 in humans suggests long-term balancing selection: there are two highly diverged haplotype clades carrying different variants of a codon that, in primates, is positively selected. For DEFB132, we show that extensive diversity, including a four-state amino acid polymorphism (valine, isoleucine, alanine and threonine at position 93), is present in hunter-gatherer populations, both African and non-African, but not found in samples from agricultural populations. CONCLUSION: Some, but not all, beta-defensin genes show positive selection in catarrhine primates. There is suggestive evidence of different selective pressures on these genes in humans, but the nature of the selective pressure remains unclear and is likely to differ between populations. PMID- 18416835 TI - New technical approach for the repair of an abdominal wall defect after a transverse rectus abdominis myocutaneous flap: a case report. AB - INTRODUCTION: Breast reconstruction with autologous tissue transfer is now a standard operation, but abnormalities of the abdominal wall contour represent a complication which has led surgeons to invent techniques to minimize the morbidity of the donor site. CASE PRESENTATION: We report the case of a woman who had bilateral transverse rectus abdominis myocutaneous flap (TRAM-flap) breast reconstruction. The surgery led to the patient developing an enormous abdominal bulge that caused her disability in terms of abdominal wall and bowel function, pain and contour. In the absence of rectus muscle, the large defect was repaired using a combination of the abdominal wall component separation technique of Ramirez et al and additional mesh augmentation with a lightweight, large-pore polypropylene mesh (Ultrapro). CONCLUSION: The procedure of Ramirez et al is helpful in achieving a tension-free closure of large defects in the anterior abdominal wall. The additional mesh augmentation allows reinforcement of the thinned lateral abdominal wall. PMID- 18416834 TI - Heterologous microarray experiments allow the identification of the early events associated with potato tuber cold sweetening. AB - BACKGROUND: Since its discovery more than 100 years ago, potato (Solanum tuberosum) tuber cold-induced sweetening (CIS) has been extensively investigated. Several carbohydrate-associated genes would seem to be involved in the process. However, many uncertainties still exist, as the relative contribution of each gene to the process is often unclear, possibly as the consequence of the heterogeneity of experimental systems. Some enzymes associated with CIS, such as beta-amylases and invertases, have still to be identified at a sequence level. In addition, little is known about the early events that trigger CIS and on the involvement/association with CIS of genes different from carbohydrate-associated genes. Many of these uncertainties could be resolved by profiling experiments, but no GeneChip is available for the potato, and the production of the potato cDNA spotted array (TIGR) has recently been discontinued. In order to obtain an overall picture of early transcriptional events associated with CIS, we investigated whether the commercially-available tomato Affymetrix GeneChip could be used to identify which potato cold-responsive gene family members should be further studied in detail by Real-Time (RT)-PCR (qPCR). RESULTS: A tomato-potato Global Match File was generated for the interpretation of various aspects of the heterologous dataset, including the retrieval of best matching potato counterparts and annotation, and the establishment of a core set of highly homologous genes. Several cold-responsive genes were identified, and their expression pattern was studied in detail by qPCR over 26 days. We detected biphasic behaviour of mRNA accumulation for carbohydrate-associated genes and our combined GeneChip-qPCR data identified, at a sequence level, enzymatic activities such as beta-amylases and invertases previously reported as being involved in CIS. The GeneChip data also unveiled important processes accompanying CIS, such as the induction of redox- and ethylene-associated genes. CONCLUSION: Our Global Match File strategy proved critical for accurately interpretating heterologous datasets, and suggests that similar approaches may be fruitful for other species. Transcript profiling of early events associated with CIS revealed a complex network of events involving sugars, redox and hormone signalling which may be either linked serially or act in parallel. The identification, at a sequence level, of various enzymes long known as having a role in CIS provides molecular tools for further understanding the phenomenon. PMID- 18416836 TI - Effects of Dielectrophoresis on Growth, Viability and Immuno-reactivity of Listeria monocytogenes. AB - Dielectrophoresis (DEP) has been regarded as a useful tool for manipulating biological cells prior to the detection of cells. Since DEP uses high AC electrical fields, it is important to examine whether these electrical fields in any way damage cells or affect their characteristics in subsequent analytical procedures. In this study, we investigated the effects of DEP manipulation on the characteristics of Listeria monocytogenes cells, including the immuno-reactivity to several Listeria-specific antibodies, the cell growth profile in liquid medium, and the cell viability on selective agar plates. It was found that a 1-h DEP treatment increased the cell immuno-reactivity to the commercial Listeria species-specific polyclonal antibodies (from KPL) by ~31.8% and to the C11E9 monoclonal antibodies by ~82.9%, whereas no significant changes were observed with either anti-InlB or anti-ActA antibodies. A 1-h DEP treatment did not cause any change in the growth profile of Listeria in the low conductive growth medium (LCGM); however, prolonged treatments (4 h or greater) caused significant delays in cell growth. The results of plating methods showed that a 4-h DEP treatment (5 MHz, 20 Vpp) reduced the viable cell numbers by 56.8-89.7 %. These results indicated that DEP manipulation may or may not affect the final detection signal in immuno-based detection depending on the type of antigen-antibody reaction involved. However, prolonged DEP treatment for manipulating bacterial cells could produce negative effects on the cell detection by growth-based methods. Careful selection of DEP operation conditions could avoid or minimize negative effects on subsequent cell detection performance. PMID- 18416837 TI - Parameter optimization in S-system models. AB - BACKGROUND: The inverse problem of identifying the topology of biological networks from their time series responses is a cornerstone challenge in systems biology. We tackle this challenge here through the parameterization of S-system models. It was previously shown that parameter identification can be performed as an optimization based on the decoupling of the differential S-system equations, which results in a set of algebraic equations. RESULTS: A novel parameterization solution is proposed for the identification of S-system models from time series when no information about the network topology is known. The method is based on eigenvector optimization of a matrix formed from multiple regression equations of the linearized decoupled S-system. Furthermore, the algorithm is extended to the optimization of network topologies with constraints on metabolites and fluxes. These constraints rejoin the system in cases where it had been fragmented by decoupling. We demonstrate with synthetic time series why the algorithm can be expected to converge in most cases. CONCLUSION: A procedure was developed that facilitates automated reverse engineering tasks for biological networks using S systems. The proposed method of eigenvector optimization constitutes an advancement over S-system parameter identification from time series using a recent method called Alternating Regression. The proposed method overcomes convergence issues encountered in alternate regression by identifying nonlinear constraints that restrict the search space to computationally feasible solutions. Because the parameter identification is still performed for each metabolite separately, the modularity and linear time characteristics of the alternating regression method are preserved. Simulation studies illustrate how the proposed algorithm identifies the correct network topology out of a collection of models which all fit the dynamical time series essentially equally well. PMID- 18416838 TI - Identification of proteins secreted by malaria parasite into erythrocyte using SVM and PSSM profiles. AB - BACKGROUND: Malaria parasite secretes various proteins in infected RBC for its growth and survival. Thus identification of these secretory proteins is important for developing vaccine/drug against malaria. The existing motif-based methods have got limited success due to lack of universal motif in all secretory proteins of malaria parasite. RESULTS: In this study a systematic attempt has been made to develop a general method for predicting secretory proteins of malaria parasite. All models were trained and tested on a non-redundant dataset of 252 secretory and 252 non-secretory proteins. We developed SVM models and achieved maximum MCC 0.72 with 85.65% accuracy and MCC 0.74 with 86.45% accuracy using amino acid and dipeptide composition respectively. SVM models were developed using split-amino acid and split-dipeptide composition and achieved maximum MCC 0.74 with 86.40% accuracy and MCC 0.77 with accuracy 88.22% respectively. In this study, for the first time PSSM profiles obtained from PSI-BLAST, have been used for predicting secretory proteins. We achieved maximum MCC 0.86 with 92.66% accuracy using PSSM based SVM model. All models developed in this study were evaluated using 5-fold cross-validation technique. CONCLUSION: This study demonstrates that secretory proteins have different residue composition than non-secretory proteins. Thus, it is possible to predict secretory proteins from its residue composition-using machine learning technique. The multiple sequence alignment provides more information than sequence itself. Thus performance of method based on PSSM profile is more accurate than method based on sequence composition. A web server PSEApred has been developed for predicting secretory proteins of malaria parasites,the URL can be found in the Availability and requirements section. PMID- 18416840 TI - Rare extracranial localization of primary intracranial neoplasm. AB - Meningioma, craniopharyngeoma and glioma are mainly intracranial lesions. Nevertheless, in rare cases these entities may occur solely as extracranial lesions that may present as intranasal/sinusoidal masses, with headaches and nasal obstruction. We present three cases of common intracranial tumors, with purely extracranial extension. The three described cases demonstrate, that preoperative MRI and CT imaging is important for differential diagnosis to exclude intracranial connections of the tumors. A definitive diagnosis requires specialized immunohistochemical examinations. In all cases of intranasal or pharyngeal neoplasm the diagnosis of meningioma, craniopharyngeoma and glioma should be considered as differential diagnosis to optimize the surgical procedure. PMID- 18416839 TI - In silico identification of conserved microRNAs in large number of diverse plant species. AB - BACKGROUND: MicroRNAs (miRNAs) are recently discovered small non-coding RNAs that play pivotal roles in gene expression, specifically at the post-transcriptional level in plants and animals. Identification of miRNAs in large number of diverse plant species is important to understand the evolution of miRNAs and miRNA targeted gene regulations. Now-a-days, publicly available databases play a central role in the in-silico biology. Because, at least ~21 miRNA families are conserved in higher plants, a homology based search using these databases can help identify orthologs or paralogs in plants. RESULTS: We searched all publicly available nucleotide databases of genome survey sequences (GSS), high-throughput genomics sequences (HTGS), expressed sequenced tags (ESTs) and nonredundant (NR) nucleotides and identified 682 miRNAs in 155 diverse plant species. We found more than 15 conserved miRNA families in 11 plant species, 10 to14 families in 10 plant species and 5 to 9 families in 29 plant species. Nineteen conserved miRNA families were identified in important model legumes such as Medicago, Lotus and soybean. Five miRNA families - miR319, miR156/157, miR169, miR165/166 and miR394 were found in 51, 45, 41, 40 and 40 diverse plant species, respectively. miR403 homologs were found in 16 dicots, whereas miR437 and miR444 homologs, as well as the miR396d/e variant of the miR396 family, were found only in monocots, thus providing large-scale authenticity for the dicot- and monocot-specific miRNAs. Furthermore, we provide computational and/or experimental evidence for the conservation of 6 newly found Arabidopsis miRNA homologs (miR158, miR391, miR824, miR825, miR827 and miR840) and 2 small RNAs (small-85 and small-87) in Brassica spp. CONCLUSION: Using all publicly available nucleotide databases, 682 miRNAs were identified in 155 diverse plant species. By combining the expression analysis with the computational approach, we found that 6 miRNAs and 2 small RNAs that have been identified only in Arabidopsis thus far, are also conserved in Brassica spp. These findings will be useful for tracing the evolution of small RNAs by examining their expression in common ancestors of the Arabidopsis Brassica lineage. PMID- 18416841 TI - "Concordance between comorbidity data from patient self-report interviews and medical record documentation". AB - BACKGROUND: Comorbidity is an important adjustment measure in research focusing on outcomes such as health status and mortality. One recurrent methodological issue concerns the concordance of comorbidity data obtained from different reporting sources. The purpose of these prospectively planned analyses was to examine the concordance of comorbidity data obtained from patient self-report survey interviews and hospital medical record documentation. METHODS: Comorbidity data were obtained using survey interviews and medical record entries from 525 hospitalized Acute Coronary Syndrome patients. Frequencies and descriptive statistics of individual and composite comorbidity data from both sources were completed. Individual item agreement was evaluated with simple and weighted kappas, Spearman Rho coefficients for composite scores. RESULTS: On average, patients reported more comorbidities during their patient survey interviews (mean = 1.78, SD = 1.99) than providers had documented in medical records (mean = 1.27, SD = 1.43). Higher proportions of positive responses were obtained from self reports compared to medical records for all conditions except congestive heart failure and renal disease. Older age and higher depressive symptom levels were significantly associated with poorer levels of data concordance. CONCLUSION: These results demonstrate that survey comorbidity data from ACS patients may not be entirely concordat with medical record documentation. In the absence of a gold standard, it is possible that hospital records did not include all pre-admission comorbidities and these patient survey interview methods may need to be refined. Self-report methods to facilitate some patients' complete recall of comorbid conditions may need to be refined by health services researchers. TRIAL REGISTRATION: ClinicalTrials.gov NCT00416026. PMID- 18416842 TI - Duplicate gene expression in allopolyploid Gossypium reveals two temporally distinct phases of expression evolution. AB - BACKGROUND: Polyploidy has played a prominent role in shaping the genomic architecture of the angiosperms. Through allopolyploidization, several modern Gossypium (cotton) species contain two divergent, although largely redundant genomes. Owing to this redundancy, these genomes can play host to an array of evolutionary processes that act on duplicate genes. RESULTS: We compared homoeolog (genes duplicated by polyploidy) contributions to the transcriptome of a natural allopolyploid and a synthetic interspecific F1 hybrid, both derived from a merger between diploid species from the Gossypium A-genome and D-genome groups. Relative levels of A- and D-genome contributions to the petal transcriptome were determined for 1,383 gene pairs. This comparison permitted partitioning of homoeolog expression biases into those arising from genomic merger and those resulting from polyploidy. Within allopolyploid Gossypium, approximately 24% of the genes with biased (unequal contributions from the two homoeologous copies) expression patterns are inferred to have arisen as a consequence of genomic merger, indicating that a substantial fraction of homoeolog expression biases occur instantaneously with hybridization. The remaining 76% of biased homoeologs reflect long-term evolutionary forces, such as duplicate gene neofunctionalization and subfunctionalization. Finally, we observed a greater number of genes biased toward the paternal D-genome and that expression biases have tended to increases during allopolyploid evolution. CONCLUSION: Our results indicate that allopolyploidization entails significant homoeolog expression modulation, both immediately as a consequence of genomic merger, and secondarily as a result of long-term evolutionary transformations in duplicate gene expression. PMID- 18416843 TI - APOE epsilon 4 lowers age at onset and is a high risk factor for Alzheimer's disease; a case control study from central Norway. AB - BACKGROUND: The objective of this study was to analyze factors influencing the risk and timing of Alzheimer's disease (AD) in central Norway. The APOE epsilon4 allele is the only consistently identified risk factor for late onset Alzheimer's disease (LOAD). We have described the allele frequencies of the apolipoprotein E gene (APOE) in a large population of patients with AD compared to the frequencies in a cognitively-normal control group, and estimated the effect of the APOE epsilon4 allele on the risk and the age at onset of AD in this population. METHODS: 376 patients diagnosed with AD and 561 cognitively-normal control individuals with no known first degree relatives with dementia were genotyped for the APOE alleles. Allele frequencies and genotypes in patients and control individuals were compared. Odds Ratio for developing AD in different genotypes was calculated. RESULTS: Odds Ratio (OR) for developing AD was significantly increased in carriers of the APOE epsilon4 allele compared to individuals with the APOE epsilon3/epsilon3 genotype. Individuals carrying APOE epsilon4/epsilon4 had OR of 12.9 for developing AD, while carriers of APOE epsilon2/epsilon4 and APOE epsilon3/epsilon4 had OR of 3.2 and 4.2 respectively. The effect of the APOE epsilon4 allele was weaker with increasing age. Carrying the APOE epsilon2 allele showed no significant protective effect against AD and did not influence age at onset of the disease. Onset in LOAD patients was significantly reduced in a dose dependent manner from 78.4 years in patients without the APOE epsilon4 allele, to 75.3 in carriers of one APOE epsilon4 allele and 72.9 in carriers of two APOE epsilon4 alleles. Age at onset in early onset AD (EOAD) was not influenced by APOE epsilon4 alleles. CONCLUSION: APOE epsilon4 is a very strong risk factor for AD in the population of central Norway, and lowers age at onset of LOAD significantly. PMID- 18416844 TI - Pleomorphic liposarcoma of the foot: a case report. AB - BACKGROUND: Liposarcomas are among the most common sarcomas of adult life. Pleomorphic liposarcoma, characterized by pleomorphic lipoblasts, is the rarest subtype. To our knowledge only three cases of pleomorphic liposarcoma of the foot or ankle have been reported so far. CASE PRESENTATION: A 71-year-old female presented with a large growing mass on the dorsum of her right foot. Computed tomography showed invasive tumorous mass. Excision biopsy revealed the mass to be a pleomorphic liposarcoma, and below the knee amputation was performed. CONCLUSION: Although the incidence of pleomorphic liposarcoma in the foot is very low, it is essential to perform thorough histological analysis of all soft tissue masses, regardless of their benign appearance, because only prompt radical surgery can result in a good prognosis for the patient. PMID- 18416845 TI - Ser80Ile mutation and a concurrent Pro25Leu variant of the VHL gene in an extended Hungarian von Hippel-Lindau family. AB - Von Hippel-Lindau disease (VHL) is a rare autosomal dominant disease characterized by development of cystic and tumorous lesions at multiple sites, including the brain, spinal cord, kidneys, adrenals, pancreas, epididymis and eyes. The clinical phenotype results from molecular abnormalities of the VHL tumor suppressor gene, mapped to human chromosome 3p25-26. The VHL gene encodes two functionally active VHL proteins due to the presence of two translational initiation sites separated by 53 codons. The majority of disease-causing mutations have been detected downstream of the second translational initiation site, but there are conflicting data as to whether few mutations located in the first 53 codons, such as the Pro25Leu could have a pathogenic role. In this paper we report a large Hungarian VHL type 2 family consisting of 32 members in whom a disease-causing AGT80AAT (Ser80Ile) c.239G>A, p.Ser80Ile mutation, but not the concurrent CCT25CTT (Pro25Leu) c.74C>T, p.Pro25Leu variant co-segregated with the disease. To our knowledge, the Ser80Ile mutation has not been previously described in VHL type 2 patients with high risk of pheochromocytoma and renal cell cancer. Therefore, this finding represents a novel genotype-phenotype association and VHL kindreds with Ser80Ile mutation will require careful surveillance for pheochromocytoma. We concluded that the Pro25Leu variant is a rare, neutral variant, but the presence such a rare gene variant may make genetic counseling difficult. PMID- 18416846 TI - Effects of intraspecific competition on the life cycle of the stonefly, Nemurella pictetii (Plecoptera: Nemouridae). AB - BACKGROUND: Considerable variation of life cycle duration in given insect species has been frequently recorded. Splitting of populations into cohorts with different life cycle lengths may occur, sometimes even between siblings from the same batch. Larval populations of the stonefly Nemurella pictetii in central Europe regularly split into a very fast developing and a normal univoltine cohort, leading to partial multivoltinism. The causes for such variation remain unknown but presumably act on the larval stage in which most of the life cycle is spent. We therefore studied possible effects of intraspecific competition on growth and development of larvae in the laboratory. RESULTS: Intraspecific competition had important influence on growth and development of the larvae. High larval densities led to reduced growth and retarded development through interference, not through exploitative competition. All specimens were negatively affected by frequent encounters and the resulting disturbance. There were no dominant individuals able to grow and develop faster than the rest, at the expense of the others. CONCLUSION: Differences in life cycle length of Nemurella pictetii may result from different larval densities in different microhabitats and resultant different degrees of interference competition. Although competition alone probably does not cause splitting of populations into cohorts with different life cycle duration differences in size and development caused by other factors are certainly enhanced by intraspecific competition. PMID- 18416847 TI - Respiratory effects of different recruitment maneuvers in acute respiratory distress syndrome. AB - INTRODUCTION: Alveolar derecruitment may occur during low tidal volume ventilation and may be prevented by recruitment maneuvers (RMs). The aim of this study was to compare two RMs in acute respiratory distress syndrome (ARDS) patients. METHODS: Nineteen patients with ARDS and protective ventilation were included in a randomized crossover study. Both RMs were applied in each patient, beginning with either continuous positive airway pressure (CPAP) with 40 cm H2O for 40 seconds or extended sigh (eSigh) consisting of a positive end-expiratory pressure maintained at 10 cm H2O above the lower inflection point of the pressure volume curve for 15 minutes. Recruited volume, arterial partial pressure of oxygen/fraction of inspired oxygen (PaO2/FiO2), and hemodynamic parameters were recorded before (baseline) and 5 and 60 minutes after RM. All patients had a lung computed tomography (CT) scan before study inclusion. RESULTS: Before RM, PaO2/FiO2 was 151 +/- 61 mm Hg. Both RMs increased oxygenation, but the increase in PaO2/FiO2 was significantly higher with eSigh than CPAP at 5 minutes (73% +/- 25% versus 44% +/- 28%; P < 0.001) and 60 minutes (68% +/- 23% versus 35% +/- 22%; P < 0.001). Only eSigh significantly increased recruited volume at 5 and 60 minutes (21% +/- 22% and 21% +/- 25%; P = 0.0003 and P = 0.001, respectively). The only difference between responders and non-responders was CT lung morphology. Eleven patients were considered as recruiters with eSigh (10 with diffuse loss of aeration) and 6 with CPAP (5 with diffuse loss of aeration). During CPAP, 2 patients needed interruption of RM due to a drop in systolic arterial pressure. CONCLUSION: Both RMs effectively increase oxygenation, but CPAP failed to increase recruited volume. When the lung is recruited with an eSigh adapted for each patient, alveolar recruitment and oxygenation are superior to those observed with CPAP. PMID- 18416848 TI - Prevalence and significance of psammoma bodies in cervicovaginal smears in a cervical cancer screening program with emphasis on a case of primary bilateral ovarian psammocarcinoma. AB - BACKGROUND: The purpose of our study was to determine the prevalence and significance of psammoma bodies (PBs) in the cervicovaginal smears of the screening population of Trento district (Italy), with the description of the cytological presentation of an asymptomatic bilateral ovarian psammocarcinoma. METHODS: From 1993 to 2006, women with PBs detected on consecutively screened cervical smears were identified from the computerized pathology database of Rovereto Hospital. The follow-up period was set from the time of cytological diagnosis to May 31st, 2007. Clinical information was obtained from retrospective review of women's medical records. The source of PBs was identified with adequate diagnostic procedures. RESULTS: PBs were found in six of the 201,231 Papanicolaou screening smears (0.0029%). Benign conditions (intrauterine device, inclusion ovarian cysts and ovarian cystoadenofibroma with PBs) were found in four patients. In two cases, PBs were associated with malignant cells; a bilateral ovarian malignancy was diagnosed in both cases, a serous adenocarcinoma and a psammocarcinoma. CONCLUSION: PBs in the cervicovaginal smears are a rare finding, associated more often with benign conditions than with malignancies. Moreover, to our knowledge, our case of primary ovarian psammocarcinoma is the first report in which the presence of malignant cells and PBs in the cervicovaginal and endometrial smears represents the first manifestation of disease. PMID- 18416849 TI - Determinants of late disease-stage presentation at diagnosis of HIV infection in Venezuela: a case-case comparison. AB - BACKGROUND: Although Venezuela has a National Human Immunodeficiency Virus (HIV) Program offering free diagnosis and treatment, 41% of patients present for diagnosis at a later disease-stage, indicating that access to care may still be limited. Our study aimed to identify factors influencing delay in presenting for HIV-diagnosis using a case-case comparison. A cross-sectional survey was performed at the Regional HIV Reference Centre (CAI), Carabobo Region, Venezuela. Between May 2005 and October 2006 225 patients diagnosed with HIV at CAI were included and demographic, behavioural and medical characteristics collected from medical files. Socio-economic and behavioural factors were obtained from 129 eligible subjects through interviews. "Late presentation" at diagnosis was defined as patients classified with disease-stage B or C according to the 1993 Centers for Disease Control and Prevention (Atlanta, USA) classification, and "early presentation" defined as diagnosis in disease-stage A. RESULTS: Of 225 subjects, 91 (40%) were defined as late presenters. A similar proportion (51/129) was obtained in the interviewed sub-sample. Older age (>30 years), male heterosexuality, lower socio-economic status, perceiving ones partner to be faithful and living >/= 25 km from the CAI were positively associated with late diagnosis in a multivariate model. Females were less likely to present late than heterosexual males (odds ratio = 0.23, P = 0.06). The main barriers to HIV testing were low knowledge of HIV/AIDS, lack of awareness of the free HIV program, lack of perceived risk of HIV-infection, fear for HIV-related stigma, fear for lack of confidentiality at testing site and logistic barriers. CONCLUSION: Despite the free Venezuelan HIV Program, poverty and barriers related to lack of knowledge and awareness of both HIV and the Program itself were important determinants in late presentation at HIV diagnosis. This study also indicates that women; heterosexual, bisexual and homosexual men might have different pathways to testing and different factors related to late presentation in each subgroup. Efforts must be directed to i) increase awareness of HIV/AIDS and the Program and ii) the identification of specific factors associated with delay in HIV diagnosis per subgroup, to help develop targeted public health interventions improving early diagnosis and prognosis of people living with HIV/AIDS in Venezuela and elsewhere. PMID- 18416850 TI - A gene sets approach for identifying prognostic gene signatures for outcome prediction. AB - BACKGROUND: Gene expression profiling is a promising approach to better estimate patient prognosis; however, there are still unresolved problems, including little overlap among similarly developed gene sets and poor performance of a developed gene set in other datasets. RESULTS: We applied a gene sets approach to develop a prognostic gene set from multiple gene expression datasets. By analyzing 12 independent breast cancer gene expression datasets comprising 1,756 tissues with 2,411 pre-defined gene sets including gene ontology categories and pathways, we found many gene sets that were prognostic in most of the analyzed datasets. Those prognostic gene sets were related to biological processes such as cell cycle and proliferation and had additional prognostic values over conventional clinical parameters such as tumor grade, lymph node status, estrogen receptor (ER) status, and tumor size. We then estimated the prediction accuracy of each gene set by performing external validation using six large datasets and identified a gene set with an average prediction accuracy of 67.55%. CONCLUSION: A gene sets approach is an effective method to develop prognostic gene sets to predict patient outcome and to understand the underlying biology of the developed gene set. Using the gene sets approach we identified many prognostic gene sets in breast cancer. PMID- 18416851 TI - Noninvasive mechanical ventilation may be useful in treating patients who fail weaning from invasive mechanical ventilation: a randomized clinical trial. AB - INTRODUCTION: The use of noninvasive positive-pressure mechanical ventilation (NPPV) has been investigated in several acute respiratory failure situations. Questions remain about its benefits when used in weaning patients from invasive mechanical ventilation (IMV). The objective of this study was to evaluate the use of bi-level NPPV for patients who fail weaning from IMV. METHODS: This experimental randomized clinical trial followed up patients undergoing IMV weaning, under ventilation for more than 48 hours, and who failed a spontaneous breathing T-piece trial. Patients with contraindications to NPPV were excluded. Before T-piece placement, arterial gases, maximal inspiratory pressure, and other parameters of IMV support were measured. During the trial, respiratory rate, tidal volume, minute volume, rapid shallow breathing index, heart rate, arterial blood pressure, and peripheral oxygen saturation were measured at 1 and 30 minutes. After failing a T-piece trial, patients were randomly divided in two groups: (a) those who were extubated and placed on NPPV and (b) those who were returned to IMV. Group results were compared using the Student t test and the chi square test. RESULTS: Of 65 patients who failed T-piece trials, 28 were placed on NPPV and 37 were placed on IMV. The ages of patients in the NPPV and IMV groups were 67.6 +/- 15.5 and 59.7 +/- 17.6 years, respectively. Heart disease, post surgery respiratory failure, and chronic pulmonary disease aggravation were the most frequent causes of IMV use. In both groups, ventilation time before T-piece trial was 7.3 +/- 4.1 days. Heart and respiratory parameters were similar for the two groups at 1 and 30 minutes of T-piece trial. The percentage of complications in the NPPV group was lower (28.6% versus 75.7%), with lower incidences of pneumonia and tracheotomy. Length of stay in the intensive care unit and mortality were not statistically different when comparing the groups. CONCLUSION: The results suggest that NPPV is a good alternative for ventilation of patients who fail initial weaning attempts. NPPV reduces the incidence of pneumonia associated with mechanical ventilation and the need for tracheotomy. TRIAL REGISTRATION: CEP HCPA (02-114). PMID- 18416852 TI - Simultaneous monitoring of independent gene expression patterns in two types of cocultured fibroblasts with different color-emitting luciferases. AB - BACKGROUND: Luciferase assay systems enable the real-time monitoring of gene expression in living cells. We have developed a dual-color luciferase assay system in which the expression of multiple genes can be tracked simultaneously using green- and red-emitting beetle luciferases. We have applied the system to monitoring independent gene expressions in two types of cocultured fibroblasts in real time. RESULTS: Two Rat-1 cell lines were established that stably express either green- or red-emitting luciferases under the control of the mBmal1 promoter, a canonical clock gene. We cocultured these cell lines, and gene expression profiles in both were monitored simultaneously. The circadian rhythms of these cell lines are independent, oscillating following their intrinsic circadian phases, even when cocultured. Furthermore, the independent rhythms were synchronized by medium change as an external stimulus. CONCLUSION: Using this system, we successfully monitored independent gene expression patterns in two lines of cocultured fibroblasts. PMID- 18416853 TI - A comparison of plotless density estimators using Monte Carlo simulation on totally enumerated field data sets. AB - BACKGROUND: Plotless density estimators are those that are based on distance measures rather than counts per unit area (quadrats or plots) to estimate the density of some usually stationary event, e.g. burrow openings, damage to plant stems, etc. These estimators typically use distance measures between events and from random points to events to derive an estimate of density. The error and bias of these estimators for the various spatial patterns found in nature have been examined using simulated populations only. In this study we investigated eight plotless density estimators to determine which were robust across a wide range of data sets from fully mapped field sites. They covered a wide range of situations including animal damage to rice and corn, nest locations, active rodent burrows and distribution of plants. Monte Carlo simulations were applied to sample the data sets, and in all cases the error of the estimate (measured as relative root mean square error) was reduced with increasing sample size. The method of calculation and ease of use in the field were also used to judge the usefulness of the estimator. Estimators were evaluated in their original published forms, although the variable area transect (VAT) and ordered distance methods have been the subjects of optimization studies. RESULTS: An estimator that was a compound of three basic distance estimators was found to be robust across all spatial patterns for sample sizes of 25 or greater. The same field methodology can be used either with the basic distance formula or the formula used with the Kendall Moran estimator in which case a reduction in error may be gained for sample sizes less than 25, however, there is no improvement for larger sample sizes. The variable area transect (VAT) method performed moderately well, is easy to use in the field, and its calculations easy to undertake. CONCLUSION: Plotless density estimators can provide an estimate of density in situations where it would not be practical to layout a plot or quadrat and can in many cases reduce the workload in the field. PMID- 18416855 TI - Brown-Vialetto-Van Laere syndrome. AB - The Brown-Vialetto-Van Laere syndrome (BVVL) is a rare neurological disorder characterized by progressive pontobulbar palsy associated with sensorineural deafness. Fifty-eight cases have been reported in just over 100 years. The female to male ratio is approximately 3:1. The age of onset of the initial symptom varies from infancy to the third decade. The syndrome most frequently presents with sensorineural deafness, which is usually progressive and severe. Lower cranial nerve involvement and lower and upper motor neuron limb signs are common neurological features. Other features include respiratory compromise (the most frequent non-neurological finding), limb weakness, slurring of speech, facial weakness, and neck and shoulder weakness. Optic atrophy, retinitis pigmentosa, macular hyperpigmentation, autonomic dysfunction, epilepsy may occur. The etiopathogenesis of the condition remains elusive. Approximately 50% of cases are familial, of which autosomal recessive is suggested. The remaining cases are sporadic. The diagnosis is usually based on the clinical presentation. Investigations (neurophysiological studies, magnetic resonance imaging of the brain, muscle biopsy, cerebrospinal fluid examination) are done to exclude other causes or to confirm the clinical findings. The differential diagnoses include the Fazio-Londe syndrome, amyotrophic lateral sclerosis, Nathalie syndrome, Boltshauser syndrome and Madras motor neuron disease. Treatment with steroids or intravenous immunoglobulin may result in temporary stabilization of the syndrome. However, the mainstays of management are supportive and symptomatic treatment, in particular assisted ventilation and maintenance of nutrition via gastrostomy. The clinical course of BVVL is variable and includes gradual deterioration (almost half of cases), gradual deterioration with stable periods in between (a third of cases) and deterioration with abrupt periods of worsening (just under a fifth of cases). After the initial presentation, one third of patients survive for ten years or longer. PMID- 18416854 TI - The underlying mechanisms for development of hypertension in the metabolic syndrome. AB - High blood pressure is an important constituent of the metabolic syndrome. However, the underlying mechanisms for development of hypertension in the metabolic syndrome are very complicated and remain still obscure. Visceral/central obesity, insulin resistance, sympathetic overactivity, oxidative stress, endothelial dysfunction, activated renin-angiotensin system, increased inflammatory mediators, and obstructive sleep apnea have been suggested to be possible factors to develop hypertension in the metabolic syndrome. Here, we will discuss how these factors influence on development of hypertension in the metabolic syndrome. PMID- 18416856 TI - Major histocompatibility complex (Mhc) class Ib gene duplications, organization and expression patterns in mouse strain C57BL/6. AB - BACKGROUND: The mouse has more than 30 Major histocompatibility complex (Mhc) class Ib genes, most of which exist in the H2 region of chromosome 17 in distinct gene clusters. Although recent progress in Mhc research has revealed the unique roles of several Mhc class Ib genes in the immune and non-immune systems, the functions of many class Ib genes have still to be elucidated. To better understand the roles of class Ib molecules, we have characterized their gene duplication, organization and expression patterns within the H2 region of the mouse strain C57BL/6. RESULTS: The genomic organization of the H2-Q, -T and -M regions was analyzed and 21 transcribed Mhc class Ib genes were identified within these regions. Dot-plot and phylogenetic analyses implied that the genes were generated by monogenic and/or multigenic duplicated events. To investigate the adult tissue, embryonic and placental expressions of these genes, we performed RT PCR gene expression profiling using gene-specific primers. Both tissue-wide and tissue-specific gene expression patterns were obtained that suggest that the variations in the gene expression may depend on the genomic location of the duplicated genes as well as locus specific mechanisms. The genes located in the H2-T region at the centromeric end of the cluster were expressed more widely than those at the telomeric end, which showed tissue-restricted expression in spite of nucleotide sequence similarities among gene paralogs. CONCLUSION: Duplicated Mhc class Ib genes located in the H2-Q, -T and -M regions are differentially expressed in a variety of developing and adult tissues. Our findings form the basis for further functional validation studies of the Mhc class Ib gene expression profiles in specific tissues, such as the brain. The duplicated gene expression results in combination with the genome analysis suggest the possibility of long-range regulation of H2-T gene expression and/or important, but as yet unidentified nucleotide changes in the promoter or enhancer regions of the genes. Since the Mhc genomic region has diversified among mouse strains, it should be a useful model region for comparative analyses of the relationships between duplicated gene organization, evolution and the regulation of expression patterns. PMID- 18416857 TI - Immunotherapy with internally inactivated virus loaded dendritic cells boosts cellular immunity but does not affect feline immunodeficiency virus infection course. AB - Immunotherapy of feline immunodeficiency virus (FIV)-infected cats with monocyte derived dendritic cells (MDCs) loaded with aldrithiol-2 (AT2)-inactivated homologous FIV was performed. Although FIV-specific lymphoproliferative responses were markedly increased, viral loads and CD4+ T cell depletion were unaffected, thus indicating that boosting antiviral cell-mediated immunity may not suffice to modify infection course appreciably. PMID- 18416859 TI - Grammaticality judgments in children: the role of age, working memory and phonological ability. AB - This paper examines the role of age, working memory span and phonological ability in the mastery of ten different grammatical constructions. Six- through eleven year-old children (n=68) and adults (n=19) performed a grammaticality judgment task as well as tests of working memory capacity and receptive phonological ability. Children showed early mastery of some grammatical structures (e.g. word order, article omissions) while even the oldest children differed from adults on others (e.g. past tense, third person singular agreement). Working memory capacity and phonological ability accounted for variance in grammaticality judgments above and beyond age effects. In particular, working memory capacity correlated with structures involving verb morphology and word order; phonological ability was important for structures with low phonetic substance. Children's relative difficulty with the different constructions showed parallels to adult performance under memory load stress, indicating working memory capacity may be a limiting factor in their performance. Implications for performance by memory and phonologically impaired populations are discussed. PMID- 18416858 TI - In patient stroke rehabilitation efficiency: influence of organization of service delivery and staff numbers. AB - BACKGROUND: Outcomes of inpatient stroke rehabilitation need to be reviewed in terms of optimal resource utilization (staff time, service organization, and duration of stay). We compared FIM efficiency scores between three hospitals, and also variation in FIM scores over a ten year period in one hospital undergoing reduction in staff numbers, to examine the relationship between outcome and service characteristics. METHOD: This is a retrospective study comparing the mean FIM efficiency for stroke patients (FIM score - FIM admission score) divided by duration of stay for 2005 among three rehabilitation hospitals adjusting for age and baseline FIM score, and a longitudinal study of changes in mean FIM efficiency during a ten year period in one hospital, to examine the effects of different service organization and staff numbers. RESULTS: FIM efficiency (FIMEG) was inversely associated with age, and positively associated with admission FIM score. FIMEG was higher in the hospital with a coordinated care plan involving medical, nursing, occupational, physiotherapy staff and other healthcare providers working as a team, with a seamless interface with community rehabilitation services. Over a ten year period, reduction in staff numbers was associated with reduction in FIMEG, which may be offset to some extent by service re-engineering. CONCLUSION: Within hospital organization of stroke rehabilitation services may influence outcome. A critical number of staff may be identified for the provision of services, below which rehabilitation efficiency may be affected. PMID- 18416860 TI - Developing spatial localization abilities and children's interpretation of where. AB - Two-year-old children often start asking questions with where. In this study we test whether children understand where to mean route or absolute location and whether the size of the space or elevation made a difference. Previous research has documented developmental changes over the preschool years in children's non verbal spatial reasoning. Forty-eight children between two and five years of age were interviewed. We asked them to point in response to where questions about an object, rooms on the same floor and on a different floor. All children pointed to the location of the hidden objects. The youngest children pointed to the route to rooms while the oldest children were more likely to point to the location of rooms. With age, the children gradually used more spatial location terms than deictic terms in response to where. These results suggest that children's meaning of where initially differs for different sized spaces and developmental changes reflect non-verbal cognition. PMID- 18416861 TI - Turkish children use morphosyntactic bootstrapping in interpreting verb meaning. AB - How might syntactic bootstrapping apply in Turkish, which employs inflectional morphology to indicate grammatical relations and allows argument ellipsis? We investigated whether Turkish speakers interpret constructions differently depending on the number of NPs in the sentence, the presence of accusative case marking and the causative morpheme. Data were collected from 60 child speakers and 16 adults. In an adaptation of Naigles, Gleitman & Gleitman (1993), the participants acted out sentences (6 transitive and 6 intransitive verbs in four different frames). The enactments were coded for causativity. Causative enactments increased in two-argument frames and decreased in one-argument frames, albeit to a lesser extent than previously found in English. This effect was generally stronger in children than in adults. Causative enactments increased when the accusative case marker was present. The causative morpheme yielded no increase in causative enactments. These findings highlight roles for morphological and syntactic cues in verb learning by Turkish children. PMID- 18416862 TI - The acquisition of German relative clauses: a case study. AB - This paper investigates the development of relative clauses in the speech of one German-speaking child aged 2 ; 0 to 5 ; 0. The earliest relative clauses we found in the data occur in topicalization constructions that are only a little different from simple sentences: they contain a single proposition, express the actor prior to other participants, assert new information and often occur with main-clause word order. In the course of the development, more complex relative constructions emerge, in which the relative clause is embedded in a fully-fledged main clause. We argue that German relative clauses develop in an incremental fashion from simple non-embedded sentences that gradually evolve into complex sentence constructions. PMID- 18416863 TI - Repetition as ratification: how parents and children place information in common ground. AB - Repetition is used for a range of functions in conversation. In this study, we examined all the repetitions used in spontaneous conversations by 41 French adult child dyads, with children aged 2 ; 3 and 3 ; 6, to test the hypotheses that adults repeat to establish that they have understood, and that children repeat to ratify what adults have said. Analysis of 978 exchanges containing repetitions showed that adults use them to check on intentions and to correct errors, while children use them to ratify what the adult said. With younger children, adults combine their repeats with new information. Children then re-repeat the form originally targeted by the adult. With older children, adults check on intentions but less frequently, and only occasionally check on forms. Older children also re repeat in the third turn but, like adults, add further information. For both adults and children, repeats signal attention to the other's utterances, and place the information repeated in common ground. PMID- 18416864 TI - Object and action picture naming in three- and five-year-old children. AB - The objectives were to explore the often reported noun advantage in children's language acquisition using a picture naming paradigm and to explore the variables that affect picture naming performance. Participants in Experiment 1 were aged three and five years, and in Experiment 2, five years. The stimuli were action and object pictures. In Experiment 1, action pictures produced more errors than object pictures for the three-year-olds, but not the five-year-olds. A qualitative analysis of the errors revealed a somewhat different pattern of errors across age groups. In Experiment 2 there was no robust difference in accuracy for the actions and objects but naming times were longer for actions. Across both experiments, imageability was a robust predictor of object naming performance, while spoken frequency was the most important predictor of action naming. The results are discussed in terms of possible differences in the manner in which nouns and verbs are acquired. PMID- 18416865 TI - Noun grammaticalization and determiner use in French children's speech: a gradual development with prosodic and lexical influences. AB - This study investigates when and how French-learning children acquire the main grammatical constraint on the noun category, i.e. the obligatory use of a preceding determiner. Spontaneous speech samples coming from the corpora of twenty children in each of three age groups, 1 ; 8, 2 ; 6, 3 ; 3, were transcribed and coded with respect to morphosyntactic, lexical and length properties of nouns. Results indicate that noun grammaticalization is a gradual process which involves early transitional procedures, as well as an increasing diversity in the content and contexts of determiner use. In support of prosodic hypotheses, noun length effects (in favor of monosyllabic nouns) mostly occurred at 1 ; 8. Animacy effects supporting the lexical hypothesis (in favor of inanimate nouns) occurred at 2 ; 6 and 3 ; 3. We suggest that noun grammaticalization is influenced by both prosodic and lexical factors. Prosodic influences predominate in the first steps of the developmental process, while lexical influences emerge in later steps. PMID- 18416866 TI - Is epenthesis a means to optimize feet? A reanalysis of the CLPF database. AB - Fikkert (1994) analyzed a large corpus of Dutch children's early language production, and found that they often add targetless syllables to their words in order to create bisyllabic feet. In this note we point out a methodological problem with that analysis: in an important number of cases, epenthetic vowels occur at places where grammatical morphemes (e.g. plural and diminutive suffixes) may be expected. Hence, the seemingly targetless syllables may represent grammatical morphemes. A reanalysis of Fikkert's original data reveals that her rhythmic explanation cannot be maintained if those cases are excluded from the analysis. PMID- 18416867 TI - The representation of morphologically complex words in the developing lexicon. AB - The study reported here examined the manner in which children represent morphologically complex words in the lexicon. Children in grades 1 to 5 completed a fragment completion task to assess the priming effects of morphologically related words. Both inflected and derived words (e.g. needs and needy, respectively) were more effective primes than control words (e.g. needle) that share similar orthography and phonology with the target word (e.g. need). These effects were consistent across the developmental period studied. Further, equivalent priming effects from the inflected and derived forms suggest that these word types are represented similarly in the developing lexicon. PMID- 18416868 TI - Acquiring causatives in Taiwan Southern Min. AB - This case study is based on the longitudinal data of a girl (LYC, 1 ; 2-3 ; 3) acquiring Taiwan Southern Min (TSM) as her first language, and it aims to discover the overgeneralization pattern of children acquiring causatives in TSM. Among the three types of causative, the errors found in other languages are mostly with lexical causatives; however, in TSM, the errors occur with morphological and analytic causatives. Being an analytic language, TSM tends to spell out the causative meaning through morphological and analytic causatives and thus most errors occur with these two types. In contrast, lexical causatives, which contain a semantic element CAUSE, were acquired late; in the data collected (1 ; 2-3 ; 3) lexical causatives were not yet found. This case study provides evidence from TSM to show a different overgeneralization pattern. PMID- 18416869 TI - Dietary pattern predicts breast cancer risk--evidence from the EPIC-Potsdam study. PMID- 18416870 TI - Nutritional profile of foods offered and consumed in a Belgian university canteen. AB - OBJECTIVE: To evaluate the nutritional profile of a lunch offered and consumed in a university canteen in Belgium. DESIGN: The qualitative and quantitative content of 4,365 meals theoretically available and 330 meals consumed was recorded during five weekdays spread over three weeks. Meal combinations were evaluated using a scoring system based on recommendations for Na content, energy from fat, and fruit and vegetable portions. SETTING: University canteen in Belgium. RESULTS: Only a 5 % of the meal combinations available and consumed complied with the three basic dietary recommendations for a hot lunch. The nutritional profile of the meals consumed was in line with that of the meals available. CONCLUSIONS: Our results show how the nutritional profile of what is eaten is largely determined by what is offered. To ensure overall compliance with dietary recommendations, considerable changes on the supply side, i.e. an increase in fruit and vegetable portions and a reduction in salt and fat of the lunch, are needed first in our setting. Our assessment provides baseline data to pilot a nutrient profiling intervention and shows how a nutrient profiling system can be used for meal evaluation purposes. PMID- 18416871 TI - Behavioral and psychological symptoms of dementia in relation to level of cognitive impairment. AB - BACKGROUND: Many people with dementia exhibit some behavioral or psychological symptoms, e.g. aggressive or aberrant motor behavior, depression or hallucinations, at some time during the course of the disorder. The aim of the present study was to describe the probability of the occurrence of these symptoms of dementia in relation to the level of cognitive impairment. METHODS: 3404 people with cognitive impairment were selected from two large cross-sectional surveys of those in geriatric care settings, conducted in 1982 and 2000 in the county of Vasterbotten, Sweden. Symptoms were assessed using the Multi Dimensional Dementia Assessment Scale (MDDAS), subsumed with a rotated factor analysis, and investigated in relation to level of cognitive impairment, measured using the Gottfries cognitive scale. RESULTS: The passiveness factor had an almost linear correlation to the level of cognitive impairment (r2 = 0.237). Non linear correlations, with highest prevalences in middle-stage cognitive impairment, were found for aggressive behavior (r2 = 0.057), wandering behavior (r2 = 0.065), restless behavior (r2 = 0.143), verbally disruptive/attention seeking behavior (r2 = 0.099), regressive/inappropriate behavior (r2 = 0.058), hallucinatory symptoms (r2 = 0.021) and depressive symptoms (r2 = 0.029). CONCLUSION: The relations between the behavioral and psychological symptoms of dementia and level of cognitive impairment were non-linear, with higher prevalence rates in the middle stages of dementia, apart from the symptom of passiveness, which increased almost linearly with the severity of cognitive impairment. PMID- 18416872 TI - Illicit drug use and problematic use in the elderly: is there a case for concern? PMID- 18416873 TI - Serum elevated gamma glutamyltransferase levels may be a marker for oxidative stress in Alzheimer's disease. AB - BACKGROUND: Gamma glutamyltransferase (GGT) plays a role in cellular glutathione uptake, which is an important element of antioxidant mechanisms. An increase in serum GGT is thought to be an early and sensitive marker of oxidative stress. Oxidative stress has a role in the pathogenesis of Alzheimer's disease (AD). The aim of this study was to investigate the GGT levels in AD. METHOD: In this cross sectional study, 132 patients with AD (mean age: 74.1 +/- 7.4, female 62.9%) and 158 age- and gender-matched normal controls (mean age: 74.5 +/- 6.3, female 67.1%) were evaluated. For cognitive assessment, MMSE and clock drawing tests were performed; DSM-IV and NINCDS-ADRDA criteria were used. Serum GGT, aspartate aminotransferase, alanine aminotransferase and alkaline phosphatase concentrations were determined. RESULTS: Median (min-max) GGT levels were 18 (9 70) in AD group and 17 (5-32) in normal controls. Mann-Whitney U test showed that GGT levels were significantly higher in AD patients (p = 0.012). Linear regression analysis revealed AD was an independent correlate of elevated GGT levels. Hypertension, diabetes mellitus, total cholesterol, and low density lipoprotein cholesterol were not associated with GGT levels. CONCLUSION: GGT levels were increased significantly in AD patients. To evaluate the role of GGT as a marker of oxidative stress in AD, further studies are needed. PMID- 18416874 TI - Research criteria for the diagnosis of Alzheimer's disease: genetic risk factors, blood biomarkers and olfactory dysfunction. PMID- 18416875 TI - The influence of drug use on fall incidents among nursing home residents: a systematic review. AB - BACKGROUND: Falls are a major health problem among the elderly, particularly in nursing homes. Abnormalities of balance and gait, psychoactive drug use, and dementia have been shown to contribute to fall risk. METHODS: We conducted a systematic review of the literature to investigate which psychoactive drugs increase fall risk and what is known about the influence of these drugs on gait in nursing home residents with dementia. We included studies with a prospective cohort design on psychoactive drug use in nursing homes with dementia residents and with falls as an outcome measure. RESULTS: Seventeen studies were included in the review. Pooled risk estimates were not calculated because there was no homogeneity across studies. We assessed the strength of evidence for psychoactive drugs as a prognostic factor for falls by defining four levels of evidence: strong, moderate, limited or inconclusive. Strong evidence was defined as consistent findings (> or =80%) in at least two high quality cohorts. We found strong evidence that the use of multiple drugs (3/3 cohorts, effect sizes 1.30-1 xs 0.30), antidepressants (10/12 cohorts, effect sizes 1.10-7.60), and anti anxiety drugs (2/2 cohorts, effect sizes 1.22-1.32) is associated with increased fall risk. The evidence for the association of other psychoactive drug classes with fall risk was limited or inconclusive. CONCLUSIONS: Research on the contribution of psychoactive drugs to fall risk in nursing home residents with dementia is limited. The scarce evidence shows, however, that multiple drugs, antidepressants and anti-anxiety drugs increase fall risk in nursing home populations with residents with dementia. PMID- 18416876 TI - The needs of people with dementia in care homes: the perspectives of users, staff and family caregivers. AB - BACKGROUND: People with dementia in care homes are at risk of having their views ignored and decisions are often made on their behalf by care home staff and family carers. The aim of this study was to compare the ratings of needs of older people with dementia living in care homes, as assessed by the older person themselves, a family caregiver, and the staff of the care home. METHOD: The Camberwell Assessment of Need for the Elderly (CANE) was completed for 238 residents with dementia by care staff. In addition, 149 users and 81 family caregivers also completed the assessment of the person's met and unmet needs. RESULTS: Many users were able to report their met and unmet needs despite having moderately severe dementia. The views of the person with dementia were commonly in agreement with the staff and family carers' views. However, user and carer pairs had the lowest average percentage agreement of 63% compared to that of 77% between user and staff pairs. In particular, users reported relatively higher unmet needs for psychological distress, company and information, and high unmet needs for daytime activities and eyesight/hearing problems. CONCLUSIONS: Users' views on their needs should be sought even when the person has dementia and lives in a care home. Reliance solely on assessment by staff or carers may lead to under recognition of unmet needs. PMID- 18416877 TI - Depressive symptoms among elderly inpatients of a Brazilian university hospital: prevalence and associated factors. AB - BACKGROUND: This study aims to evaluate the prevalence of depressive symptoms and associated factors among elderly hospital inpatients. METHODS: A cross-sectional study evaluated 189 participants using the Geriatric Depression Scale, the Mini mental State Examination and the Katz and Lawton Index, to assess dependence regarding activities of daily living (ADL). RESULTS: Most of the participants were women, aged between 60 and 92 years, with low levels of educational attainment and personal income, and non-qualified occupations. The prevalence of depressive symptoms was 56%, but only 3% had a psychiatric diagnosis registered in their medical records. Univariate analysis showed significant associations between depressive symptoms and low educational level and income, marital status, number of hospitalizations in the previous year, cognitive decline, dependence regarding basic and instrumental ADL, and death. After logistic regression, the only variables that remained significantly associated with depression were low educational level, dependence regarding basic ADL, and death. CONCLUSIONS: Depressive symptoms were independently associated with low educational level and dependence regarding basic ADL. Hospitalized elderly people with depressive symptoms were more likely to die. It is essential to diagnose and treat depression properly in this population to minimize its negative impacts. PMID- 18416878 TI - Facial nerve palsy secondary to middle-ear lipoma. AB - OBJECTIVE: We present the first reported case of a middle-ear lipoma presenting with facial nerve palsy. We review the available literature on middle-ear lipomas and alert the surgeon to the possibility of a lipoma occurring in this location. CASE REPORT: A 33-year-old man presented to our unit with a right-sided, House Brackmann grade two, lower motor neurone facial palsy. A computed tomography scan revealed abnormal soft tissue in the epitympanic recess, extending to the region of the geniculate ganglion. At middle-ear exploration, a lump of fatty tissue was found filling the anterior middle-ear cleft, juxtaposed to the horizontal portion of the facial nerve. The patient's facial palsy resolved within a few weeks of surgery. CONCLUSION: Lipomas are a rare but real differential diagnosis of a mass in the middle ear. Early imaging is advised. PMID- 18416879 TI - Deafness in the developing world: the place of cochlear implantation. AB - INTRODUCTION: This paper attempts to review changes in the lives of hearing impaired patients within the developing world, brought about by globalisation and development. The paper also explores limitations to improved care and addresses the collective moral responsibility of developed nations. METHODS: Analysis of literature. RESULTS: Within developing nations, large populations have emerged with a similar pattern of problems, access to information and aspirations as those living in developed nations. However, marked differences in income have persisted. These trends have resulted in a relative increase in the proportion of the hearing-impaired population in need of cochlear implantation, while at the same time restricting their access to such treatment. CONCLUSIONS: The emergence of global markets and media and a shared sense of destiny amongst the people of this planet should translate into a concerted, worldwide effort to assist the deaf in developing countries. Much more can be done within existing resources and frameworks to improve the quality of these peoples' lives. PMID- 18416880 TI - Endoscopic modified Lothrop procedure for repair of lateral frontal sinus cerebrospinal fluid leak. AB - OBJECTIVE: To describe a previously unreported case of repair of a lateral frontal sinus cerebrospinal fluid leak, using the endoscopic modified Lothrop procedure. METHOD: Case report of new technique, with reference to the world literature. RESULTS: An effective endoscopic, transnasal repair of a lateral frontal sinus cerebrospinal fluid leak was achieved in a 60-year-old man. The defect was closed with fat, fascia lata and free mucosal grafts. The left nasal cavity was packed and a lumbar drain left in situ post-operatively. The drain and packs were removed one week later and the patient discharged with no complications, and no recurrence at 12 months' follow up. CONCLUSION: Such cerebrospinal fluid leaks have traditionally been repaired using an external approach with osteoplastic flaps and obliteration of the sinus. We highlight the endoscopic modified Lothrop technique as an effective alternative approach to repair of cerebrospinal fluid leaks in poorly accessible areas of the frontal sinus. PMID- 18416881 TI - Strongyloides stercoralis: a field-based survey of mothers and their preschool children using ELISA, Baermann and Koga plate methods reveals low endemicity in western Uganda. AB - To ascertain the current status of strongyloidiasis in mothers and their preschool children, a field-based survey was conducted in western Uganda using a combination of diagnostic methods: ELISA, Baermann concentration and Koga agar plate. The prevalence of other soil-transmitted helminthiasis and intestinal schistosomiasis were also determined. In total, 158 mothers and 143 children were examined from five villages within Kabale, Hoima and Masindi districts. In mothers and children, the general prevalence of strongyloidiasis inferred by ELISA was approximately 4% and approximately 2%, respectively. Using the Baermann concentration method, two parasitologically proven cases were encountered in an unrelated mother and child, both of whom were sero-negative for strongyloidiasis. No infections were detected by Koga agar plate method. The general level of awareness of strongyloidiasis was very poor ( < 5%) in comparison to schistosomiasis (51%) and ascariasis (36%). Strongyloidiasis is presently at insufficient levels to justify inclusion within a community treatment programme targeting maternal and child health. Better epidemiological screening is needed, however, especially identifying infections in HIV-positive women of childbearing age. In the rural clinic setting, further use of the Baermann concentration method would appear to be the most immediate and pragmatic option for disease diagnosis. PMID- 18416883 TI - Larval migration of Toxocara canis in piglets and transfer of larvae from infected porcine tissue to mice. AB - Visceral larva migrans (VLM), caused by Toxocara canis larvae in humans, animals and birds, is now well documented throughout the world. Seven piglets were infected orally with 5 x 104 embryonated eggs and the migration and distribution of T. canis larvae in the tissues were evaluated. After artificial gastric juice digestion, larval yields at necropsy from different organs and muscles on days 1, 3, 7, 15 and 30 post-infection (DPI) revealed 3.05, 0.97, 0.21, 0.13, 0.05, 0.14% recovery from liver, lungs, heart, kidneys, skeletal muscles and brain tissues respectively, with a total of 2486 (4.97%) recovery from all tissues together. The highest number of larvae 1527 (3.05%) was recovered from the liver throughout the period (1-30 DPI), indicating a special affinity of larvae for the liver. Subsequently five mice were each infected orally with 5 g of infected pig liver and, after necropsy on 10 DPI, 20 +/- 3.62, 17 +/- 5.10, 3 +/- 1.26, 12 +/- 3.92 and 30 +/- 5.69 larvae were recovered from liver, lungs, heart, brain and muscles, respectively. Thus, primarily, the migratory potential and adaptation of T. canis larvae in porcine tissue was examined and, subsequently, their establishment in the second paratenic host, the mouse, has been successful. No influence of host sex on the migratory potential of T. canis larvae was observed. The related pathology caused by migratory larvae and its zoonotic significance through the consumption of raw or undercooked pork has been emphasized. PMID- 18416882 TI - Minimum sampling effort for reliable non-invasive estimations of excretion abundance of Elaphostrongylus cervi L1 in red deer (Cervus elaphus) populations. AB - Faecal surveys are commonly used as non-invasive means to evaluate population abundance of animals as well as comparable indexes of prevalence and intensity of diseases between populations, especially macroparasites. While faecal surveys are among one of the simplest means to perform these evaluations, they are time consuming and labour intensive. The present study evaluated 80 red deer (Cervus elaphus) faecal samples collected in two study sites for the presence and abundance of first-stage larvae of the nematode Elaphostrongylus cervi and established pools of samples for epidemiological analysis. The analysis of 20-30 individual samples would produce a reliable estimate of the 'true' prevalence, and the error of the smaller sample size only doubled that of the 80 reference samples. The analysis of 5 pools of 5 pellet groups each, or of 4 pools of 10 pellet groups each, provided a reliable estimation of the 'true' excretion abundance estimated by the analysis of all 80 samples. These pools comprise a cost-effective and dependable alternative for measuring and obtaining comparable abundances of parasitic faecal excretion stages in red deer populations, which is very valuable for epidemiological and ecological research as well as for management purposes. PMID- 18416884 TI - Chondroitin sulphate for symptomatic osteoarthritis: critical appraisal of meta analyses. AB - BACKGROUND: Chondroitin sulphate (CS) is an important structural component of cartilage and is approved and regulated as a symptomatic slow-acting drug for osteoarthritis (OA) (SYSADOA) in Europe and some other countries. Although numerous studies have shown the clinical benefits of CS to decrease pain, improve functional disability, reduce non-steroidal anti-inflammatory drug (NSAID) or acetaminophen consumption, and good tolerability with an additional carry-over effect, there are still some concerns regarding its effectiveness in treating OA. PURPOSE: To examine the data provided by meta-analyses to clarify the effectiveness of CS as a symptomatic treatment for OA. METHODS: A MEDLINE database search was conducted for appropriate meta-analyses published between 1997 and 2007. Five meta-analyses that limited their analysis to randomised controlled trials (RCTs) comparing CS with placebo or no-treatment control arms were retrieved. RESULTS: Four meta-analyses showed significant clinical effects of CS compared with placebo for pain and function measures and one demonstrated greater reduction of analgesic co-medication in patients assigned to the active treatment. In one meta-analysis, the 20 trials included in the study showed a high degree of heterogeneity and the conclusion that CS showed minimal symptomatic benefits was based on the analysis of only three trials. One meta analysis showed that pain relief after CS treatment steadily increased between 4 and 12 weeks of treatment, whereas the time course of pain relief after treatment with NSAIDs decreased. Two meta-analyses reported consistently higher frequencies of side effects in the placebo group than in patients treated with CS. CONCLUSION: Data provided by these meta-analyses indicate that CS has a slight to moderate efficacy in the symptomatic treatment of OA, with an excellent safety profile. PMID- 18416885 TI - 24-Week study on the use of collagen hydrolysate as a dietary supplement in athletes with activity-related joint pain. AB - BACKGROUND: Collagen hydrolysate is a nutritional supplement that has been shown to exert an anabolic effect on cartilage tissue. Its administration appears beneficial in patients with osteoarthritis. OBJECTIVE: To investigate the effect of collagen hydrolysate on activity-related joint pain in athletes who are physically active and have no evidence of joint disease. DESIGN AND SETTING: A prospective, randomized, placebo-controlled, double-blind study was conducted at Penn State University in University Park, Pennsylvania. Parameters including joint pain, mobility, and inflammation were evaluated with the use of a visual analogue scale during a 24-week study phase. STUDY PARTICIPANTS: Between September 2005 and June 2006, 147 subjects who competed on a varsity team or a club sport were recruited. Data from 97 of 147 subjects could be statistically evaluated. INTERVENTION: One hundred and forty-seven subjects (72 male, 75 female) were randomly assigned to two groups: a group (n = 73) receiving 25 mL of a liquid formulation that contained 10 g of collagen hydrolysate (CH-Alpha) and a group (n = 74) receiving a placebo, which consisted of 25 mL of liquid that contained xanthan. MAIN OUTCOME MEASURES: The primary efficacy parameter was the change in the visual analogue scales from baseline during the study phase in relation to the parameters referring to pain, mobility, and inflammation. RESULTS: When data from all subjects (n = 97) were evaluated, six parameters showed statistically significant changes with the dietary supplement collagen hydrolysate (CH) compared with placebo: joint pain at rest, assessed by the physician (CH vs. placebo (-1.37 +/- 1.78 vs. -0.90 +/- 1.74 (p = 0.025)) and five parameters assessed by study participants: joint pain when walking (-1.11 +/ 1.98 vs. -0.46 +/- 1.63, p = 0.007), joint pain when standing (-0.97 +/- 1.92 vs. -0.43 +/- 1.74, p = 0.011), joint pain at rest (-0.81 +/- 1.77 vs. -0.39 +/- 1.56, p = 0.039), joint pain when carrying objects (-1.45 +/- 2.11 vs. -0.83 +/- 1.71, p = 0.014) and joint pain when lifting (-1.79 +/- 2.11 vs. -1.26 +/- 2.09, p = 0.018). When a subgroup analysis of subjects with knee arthralgia (n = 63) was performed, the difference between the effect of collagen hydrolysate vs. placebo was more pronounced. The parameter joint pain at rest, assessed by the physician, had a statistical significance level of p = 0.001 (-1.67 +/- 1.89 vs. 0.86 +/- 1.77), while the other five parameters based on the participants' assessments were also statistically significant: joint pain when walking (p = 0.003 (-1.38 +/- 2.12 vs. -0.54 +/- 1.65)), joint pain when standing (p = 0.015 ( 1.17 +/- 2.06 vs. -0.50 +/- 1.68)), joint pain at rest with (p = 0.021 (-1.01 +/ 1.92 vs. -0.47 +/- 1.63)), joint pain when running a straight line (p = 0.027 ( 1.50 +/- 1.97 vs. -0.80 +/- 1.66)) and joint pain when changing direction (p = 0.026 (-1.87 +/- 2.18 vs. -1.20 +/- 2.10)). CONCLUSION: This was the first clinical trial of 24-weeks duration to show improvement of joint pain in athletes who were treated with the dietary supplement collagen hydrolysate. The results of this study have implications for the use of collagen hydrolysate to support joint health and possibly reduce the risk of joint deterioration in a high-risk group. Despite the study's size and limitations, the results suggest that athletes consuming collagen hydrolysate can reduce parameters (such as pain) that have a negative impact on athletic performance. Future studies are needed to support these findings. PMID- 18416886 TI - Cost-effectiveness of escitalopram for generalized anxiety disorder in Canada. AB - BACKGROUND: Generalized Anxiety Disorder (GAD) is a common chronic disease with a lifetime prevalence estimated to range from 4.2% to 12.7%. GAD places a substantial burden upon patients and healthcare resources. OBJECTIVE: To determine the cost-effectiveness of escitalopram for GAD in a Canadian primary care setting from two perspectives [Ministry of Health (MoH) and society (SOC)]. METHODS: A 24-week decision-analytic model was constructed using Data/TreeAge software. Patients were treated with escitalopram or generic paroxetine. Clinical rates were determined from the literature; expert opinion guided model pathway development. Effectiveness was measured as 'symptom-free days' (SFDs). Analyses from MoH perspective focused on direct costs of treatment (drugs, physician visits), while SOC also accounted for indirect costs associated with workdays lost due to GAD. Unit costs of healthcare services and wage rates were obtained from standard Canadian sources (2005 Canadian $ values). Cost-effectiveness was expressed as the incremental cost-effectiveness ratio (ICER). Extensive one-way and probabilistic sensitivity analyses were conducted. RESULTS: Escitalopram was associated with higher expected number of SFDs than paroxetine (86.4 vs. 77.0 SFD, respectively). From the MoH perspective, expected costs were Can$724 and Can$663 for escitalopram and paroxetine arms, respectively, resulting in the ICER for escitalopram vs. paroxetine of Can$6.56/SFD (Can$2362/symptom free year). From the SOC perspective, escitalopram dominated paroxetine as more effective on SFDs and less costly. Sensitivity analyses demonstrated robustness of the model. Limitations include the absence of comorbidities, which are common in practice, lack of long-term data, and assuming that dropouts in trials reflect those in practice. CONCLUSION: Escitalopram was found to be cost-effective compared with paroxetine in treatment of GAD from the Canadian MoH perspective, and dominating paroxetine from the SOC perspective. Therefore, a possible advantage may exist at the population level in the treatment of GAD with escitalopram in Canada. PMID- 18416890 TI - Photophysics of 9,10-anthracenediol and a bifunctional sacrificial template in solution and xerogels. AB - Site selectively templated and tagged xerogels (SSTTX) represent a new sensing platform. Although this platform has several attractive features, the template formation process is not fully understood. To address this issue we have explored the photophysics of two model compounds (9,10-anthracenediol and a bifunctional sacrificial template (BST)) when dissolved in solution and when sequestered within a xerogel. The solution experiments show that the carbamate tethers on the BST (which are eventually cleaved to form the analyte responsive sites that make up the SSTTX) do not alter the anthracene residue's intrinsic photophysics. In contrast, 9,10-anthracenediol and BST molecules sequestered within a xerogel sense and report from a distribution of microenvironments. The distribution mean values are very similar, but the variance is statistically greater for the BST doped xerogel in comparison to the 9,10-anthracenediol-doped xerogel. The most likely causes of this behavior are heterogeneity and electron and energy transfer processes that are controlled by differences in the position/orientation of the anthracene moiety at the pore surface in the Class I (9,10-anthracenediol) and Class II (BST) xerogels. These results also suggest that the initial template sites produced during the SSTTX formation process are not discrete; they are intrinsically more diverse (maybe 30%) in comparison to the types of template sites created by traditional molecular imprinting strategies. However, our previously reported SSTTX binding studies do not reveal any evidence for a distribution of analyte-to-SSTTX binding. This apparently anomalous behavior may result because the relative standard deviation of the binding process is intrinsically small and/or one or more of the steps that follow template site formation attenuate the final template site distribution within the SSTTX. PMID- 18416887 TI - Direct and indirect costs among employees with diabetic retinopathy in the United States. AB - OBJECTIVE: To examine, from the employer perspective, the direct (healthcare) and indirect (workloss) costs of employees with diabetic retinopathy (DR) compared to control non-DR employees with diabetes, and within DR subgroups. METHODS: Compared annual costs using claims data from 17 large companies (1999-2004). 'DR employees' (n = 2098) had >or= 1 DR (International Classification of Disease, 9th Revision [ICD-9]) diagnosis; DR subgroups included employees with diabetic macular edema (DME), proliferative DR (PDR), and employees receiving photocoagulation or vitrectomy procedures. Descriptive and multivariate tests were performed. RESULTS: DR employee annual direct costs were $18,218 (indirect = $3548) compared to $11,898 (indirect = $2374) for controls (Delta = $2032 (adjusted); p < 0.0001). Costs differences were larger across DR employee subgroups: DME/non-DME ($28,606/$16,363); PDR/non-PDR ($30,135/$13,445; p < 0.0001); DR with/without photocoagulation ($34,539/$16,041; p < 0.0001); and DR with/without vitrectomy ($63,933/$17,239; p < 0.0001). LIMITATIONS: This study examined the incremental costs of treating DR employees, which may be higher than the incremental costs of DR itself. Some measures of diabetes severity (e.g., duration of diabetes) were not available in the claims data, and were therefore not included in the multivariate models. The cost of photocoagulation and vitrectomy procedures pertain to individuals who underwent these procedures, and not the cost of the procedures themselves. CONCLUSION: DR employees had significantly higher costs than controls, and larger differences existed within DR subgroups. Indirect costs accounted for about 20% of total cost. PMID- 18416891 TI - Standoff detection of chemical and biological threats using laser-induced breakdown spectroscopy. AB - Laser-induced breakdown spectroscopy (LIBS) is a promising technique for real time chemical and biological warfare agent detection in the field. We have demonstrated the detection and discrimination of the biological warfare agent surrogates Bacillus subtilis (BG) (2% false negatives, 0% false positives) and ovalbumin (0% false negatives, 1% false positives) at 20 meters using standoff laser-induced breakdown spectroscopy (ST-LIBS) and linear correlation. Unknown interferent samples (not included in the model), samples on different substrates, and mixtures of BG and Arizona road dust have been classified with reasonable success using partial least squares discriminant analysis (PLS-DA). A few of the samples tested such as the soot (not included in the model) and the 25% BG:75% dust mixture resulted in a significant number of false positives or false negatives, respectively. Our preliminary results indicate that while LIBS is able to discriminate biomaterials with similar elemental compositions at standoff distances based on differences in key intensity ratios, further work is needed to reduce the number of false positives/negatives by refining the PLS-DA model to include a sufficient range of material classes and carefully selecting a detection threshold. In addition, we have demonstrated that LIBS can distinguish five different organophosphate nerve agent simulants at 20 meters, despite their similar stoichiometric formulas. Finally, a combined PLS-DA model for chemical, biological, and explosives detection using a single ST-LIBS sensor has been developed in order to demonstrate the potential of standoff LIBS for universal hazardous materials detection. PMID- 18416892 TI - Raman signal processing software for automated identification of mineral phases and biosignatures on Mars. AB - Data from the ESA ExoMars Rover Mission will provide invaluable input for further studies in astro/exobiology. The search for mineral products as indicators of present and/or past biogenetic activities in Mars' surface and subsurface samples is the main objective of the compact Raman-laser-induced breakdown spectroscopy (LIBS) instrument. The inherent features of Raman spectroscopy and LIBS make the combined instrument a unique and very powerful tool in the search for biomarkers and hence it is regarded as the highest priority instrument for mineral analysis within the mission. We have developed a software package for the on-board processing of the instrument's data outputs, including spectral conditioning and search-match characterization of mineral phases and biomarkers. In this paper we show the mathematical and physical basis of the software package. PMID- 18416893 TI - Surface-enhanced Raman scattering spectroscopy as a sensitive and selective technique for the detection of folic acid in water and human serum. AB - Surface-enhanced Raman scattering (SERS) is shown to give linear and sensitive concentration-dependent detection of folic acid using silver nanoparticles created via ethylene-diaminetetraacetic acid (EDTA) reduction. Optical detection by SERS overcomes the primary limitation of photodissociation encountered during the application of other shorter wavelength ultraviolet (UV)/near-UV techniques such as fluorescence based microscopy. The SERS approach in water-based samples was demonstrated and optimized using several longer wavelengths of excitation (514.5, 632.8, and 785 nm). Excitation in the green (514.5 nm) was found to achieve the best balance between photodissociation and SERS efficiency. Linear concentration dependence was observed in the range of 0.018 to 1 microM. The importance of folic acid in a clinical setting and the potential applications of this technique in a biological environment are highlighted. We demonstrate the potential to transfer this technique to real biological samples by the detection of folic acid in human serum samples by SERS. PMID- 18416894 TI - Mid-infrared diffuse reflection of a strongly absorbing analyte on non-absorbing and absorbing matrices. Part I: homogeneous powders. AB - KBr disks of caffeine were prepared quantitatively so that the absorptivity of caffeine in this environment could be measured. The disks were then ground up finely and their diffuse reflection spectrum was measured. The average path length traveled by remitted photons was then calculated by the simple application of the Beer-Lambert law. The path length that an average photon travels within a finely powdered matrix of KBr is approximately 4 mm when the concentration of a uniformly dispersed, strongly absorbing analyte such as caffeine is 0.01% or less. This path length then decreases as the concentration of the analyte increases as more photons are absorbed by the analyte. When carbon black is added to the mixture of caffeine and KBr and the measurements are repeated, the effective path length drops even further. The average photon that is remitted from a fine infrared-transparent powder containing a very low concentration of an analyte has been shown to encounter at least 400 particles, indicating a highly random path. The more strongly the matrix absorbs, the shorter the path length. When 0.1% of carbon black is added to the disk, the path length drops to about 100 microm. PMID- 18416895 TI - Mid-infrared diffuse reflection of a strongly absorbing analyte on non-absorbing and absorbing matrices. Part II: thin liquid layers on powdered substrates. AB - The average total path length traveled by photons in mid-infrared diffuse reflection spectrometry of powdered potassium chloride matrices coated with a thin layer of silicone oil has been estimated. The average path length of the photons that had passed through the silicone oil layer was first calculated by the application of the Beer-Lambert law. The total path length through the coated particles was then estimated by dividing the path length of the silicone oil by its concentration. The average path length of the photons that had been diffusely reflected from KCl particles with an average diameter of approximately 5 microm that had been coated with a very thin layer of the silicone oil (corresponding to a concentration of 0.01% w:w) was estimated to be about 8 mm. This path length was significantly reduced either as the silicone oil concentration or the absorption of the matrix was increased. The result helps to explain why diffuse reflection spectrometry is a far poorer sampling technique for trace analysis when the matrix has strong absorption and why diffuse reflection is not a very promising technique for the standoff detection of nonvolatile chemical warfare agents. PMID- 18416896 TI - Sequential wavelength tuning: dynamics at interfaces investigated by vibrational sum-frequency spectroscopy. AB - Vibrational sum-frequency spectroscopy is a powerful tool for the study of interfaces, but its application has hitherto mainly been limited to static structure. This contribution demonstrates how the considerably improved stability of state-of-the-art lasers and parametric generators can be exploited to study the evolution of interfacial structure continuously for several hours. By sequential wavelength tuning and automated control of spatial beam overlap at the target, amplitude changes of sum-frequency resonances in widely spaced infrared regions can be probed. This offers great advantages for the study of the synchronism of molecular processes at interfaces. PMID- 18416897 TI - Pulsed terahertz attenuated total reflection spectroscopy. AB - Pulsed terahertz attenuated total reflection (ATR) spectra of solid materials and liquids covering the 10 cm(-1) to 120 cm(-1) (0.3 THz to 3.6 THz) region of the electromagnetic spectrum are recorded using a terahertz pulsed spectrometer and silicon ATR modules. Pulsed terahertz ATR measurements are completed nondestructively using small amounts of sample (typically 1 mg for solids) and no sample preparation. Many terahertz analyses can be run in rapid sequence, minimizing the analysis time. PMID- 18416898 TI - Estimation of effective refractive index of birefringent particles using a combination of the immersion liquid method and light scattering. AB - A method to detect the effective refractive index and concentration of birefringent pigments is suggested. The method is based on the utilization of the immersion liquid method and a multifunction spectrophotometer for the measurement of back scattered light. The method has applications in the measurement of the effective refractive index of pigments that are used, e.g., in the paper industry to improve the opacity of paper products. PMID- 18416899 TI - Novel spectropolarimeter employing fixed polarizers for the determination of optically active samples. AB - A novel spectropolarimeter, based on modification of an ordinary, inexpensive, multiwavelength ultraviolet (UV)-visible-near-infrared (NIR) spectrophotometer, is described and applied to the determination of sucrose, sucrose inversion, and enantiomeric composition of solutions of (R)-(+)-limonene and (S)-(-)-limonene. The instrument has no moving parts, and optical rotation measurements are encoded as an apparent absorbance. Apparent absorbance measurements can be combined with multivariate statistical analysis over a wide spectral range, and a background correction technique that employs the sample as its own blank provides an effective means of correcting for the presence of chromophores that also absorb over the wavelengths of interest. The instrument was tested against an ordinary polarimeter and showed good performance with both colorless and colored samples. PMID- 18416900 TI - Prediction of long and short residue properties of crude oils from their infrared and near-infrared spectra. AB - Research has been carried out to determine the feasibility of chemometric modeling of infrared (IR) and near-infrared (NIR) spectra of crude oils to predict the long residue (LR) and short residue (SR) properties of these samples. A novel method is described to predict short residue properties at different flashing temperatures based on the IR spectrum of a crude oil measured at room temperature. The resulting method is the subject of European patent application number 07251853.3 filed by Shell Internationale Research Maatschappij B.V. The study has been carried out on 47 crude oils and 4 blends, representing a large variety of physical and chemical properties. From this set, 28 representative samples were selected by principle component analysis (PCA) and used for calibration. The remaining 23 samples were used as a test set to validate the obtained partial least squares (PLS) regression models. The results demonstrate that this integrated approach offers a fast and viable alternative for the currently applied elaborate ASTM (American Society for Testing and Materials) and IP (Institute of Petroleum) methods. IR spectra, in particular, were found to be a useful input for the prediction of different LR properties. Root mean square error of prediction values of the same order of magnitude as the reproducibility values of the ASTM methods were obtained for yield long on crude (YLC), density (D(LR)), viscosity (V(LR)), and pour point (PP), while the ability to predict the sulfur contents (S) and the carbon residue (CR) was found to be useful for indicative purposes. The prediction of SR properties is also promising. Modeling of the IR spectra, and to a lesser extent, the NIR spectra as a function of the average flash temperature (AFT) was particularly successful for the prediction of the short residue properties density (D(SR)) and viscosity (V(SR)). Similar results were obtained from the models to predict SR properties as a function of the yield short on crude (YSC) values. Finally, it was concluded that the applied protocol including sample pretreatment and instrumental measurement is highly reproducible and instrument and accessory independent. PMID- 18416901 TI - Novel near-infrared sampling apparatus for single kernel analysis of oil content in maize. AB - A method of rapid, nondestructive chemical and physical analysis of individual maize (Zea mays L.) kernels is needed for the development of high value food, feed, and fuel traits. Near-infrared (NIR) spectroscopy offers a robust nondestructive method of trait determination. However, traditional NIR bulk sampling techniques cannot be applied successfully to individual kernels. Obtaining optimized single kernel NIR spectra for applied chemometric predictive analysis requires a novel sampling technique that can account for the heterogeneous forms, morphologies, and opacities exhibited in individual maize kernels. In this study such a novel technique is described and compared to less effective means of single kernel NIR analysis. Results of the application of a partial least squares (PLS) derived model for predictive determination of percent oil content per individual kernel are shown. PMID- 18416902 TI - Near-infrared analysis of whole kernel barley: comparison of three spectrometers. AB - This study was conducted to develop calibration models for determining quality parameters of whole kernel barley using a rapid and nondestructive near-infrared (NIR) spectroscopic method. Two hundred and five samples of whole barley grains of three winter-habit types (hulled, malt, and hull-less) produced over three growing seasons and from various locations in the United States were used in this study. Among these samples, 137 were used for calibration and 68 for validation. Three NIR instruments with different resolutions, one Fourier transform instrument (4 cm(-1) resolution), and two dispersive instruments (8 nm and 10 nm bandpass) were utilized to develop calibration models for six components (moisture, starch, beta-glucan, protein, oil, and ash) and the results were compared. Partial least squares regression was used to build models, and various methods for preprocessing of spectral data were used to find the best model. Our results reveal that the coefficient of determination for calibration models (NIR predicted versus reference values) ranged from 0.96 for moisture to 0.79 for beta glucan. The level of precision of the model developed for each component was sufficient for screening or classification of whole kernel barley, except for beta-glucan. The higher resolution Fourier transform instrument gave better results than the lower resolution instrument for starch and beta-glucan analysis. The starch model was most improved by the increased resolution. There was no advantage of using a higher resolution instrument over a lower resolution instrument for other components. Most of the components were best predicted using first-derivative processing, except for beta-glucan, where second-derivative processing was more informative and precise. PMID- 18416903 TI - Laser-induced fluorescence ratios of Cajanus cajan L. under the stress of cadmium and its correlation with pigment content and pigment ratios. AB - Laser-induced fluorescence spectra were used to characterize the effect of cadmium on the pigment status of the leaves of Cajanus cajan L. Laser-induced fluorescence spectra of untreated as well as cadmium treated (0.01 mM, 0.10 mM, and 1.00 mM) Cajanus cajan L. were recorded using the 355 nm line of a Nd:YAG laser as the excitation source and a monochromator with an intensified charge coupled device as a detector in the region 400-800 nm. The fluorescence intensity ratios (FIR) of control as well as treated Cajanus cajan L. have been calculated by evaluating curve fitted parameters using a Gaussian spectral function. In addition, some growth parameters, such as photosynthetic pigment content, were also measured. The 355 nm line of the laser-light-excited leaves not only showed a fluorescence emission in the red spectral region (650-800 nm), but also in the blue-green region (400-570 nm). The chlorophyll FIR F690/F740 strongly correlated with the photosynthetic pigment content (total chlorophyll and carotenoids) and its ratio. Consequently, a correlation was also seen between the ratio of the blue-green fluorescence F470/F540 and the photosynthetic pigment content. The results indicated that the plants treated with 0.01 mM of cadmium exhibited better growth, while higher concentrations of cadmium were hazardous for Cajanus cajan L. PMID- 18416904 TI - Mode-mismatched dual-beam differential thermal lensing with optical scheme design optimized using expert estimation for analytical measurements. AB - The optimization of the optical scheme design of a mode-mismatched dual-beam thermal-lens spectrometer for differential (dual-cell) measurements in a far field mode using diffraction thermal-lens theory is carried out. A criterion for an expert estimation of the quality of the spectrometer design for differential thermal-lens measurements in analytical chemistry (sensitivity, low limits of detection, and quantification) is also developed. The theoretical calculations agree well with previous papers on differential thermal lensing. Using the example of iron(II) tris-(1,10-phenanthrolinate), it is shown that the blank signal compensation in differential thermal lens spectrometry provides a decrease in the limit of detection by an order of magnitude compared to the decrease in single-cell measurements. Using an artificial two-component mixture of ferroin and potassium dichromate, it is shown that dual-beam differential thermal lens spectrometry makes it possible to determine trace components against 900-fold excess amounts of interfering substances. PMID- 18416905 TI - Photothermal deflection determination of iron(II) with ferrozine with sorption preconcentration on Silufol plates. AB - Photothermal deflection spectroscopy was applied to the selective detection of iron(II) chelate with ferrozine by its sorption preconcentration on Silufol plates. The linearity range was 1 x 10(-11) - 6 x 10(-8) mol cm(-2) of chelate at the plate surface, which corresponded to 1 x 10(-9) -4 x 10(-6) M of chelate in solution. The limits of detection and quantification are 8 x 10(-12) and 2.5 x 10(-11) mol cm(-2) at the plate from 15 microL of test solution (0.5 nM and 1.5 nM in solution, respectively), and the absolute detection limit is 8 fmol in the whole spot applied to a plate. Characteristics and features of photothermal deflection detection are discussed. PMID- 18416906 TI - Laser-induced breakdown spectroscopy for determination of the organic oxygen content in anthracite coal under atmospheric conditions. AB - Laser-induced breakdown spectroscopy has been used to measure the organic oxygen content in pulverized anthracite coal under atmospheric conditions. Special spectral processing including the optimal O(I) emission-line selection by comparing the spectral correlation coefficients with the N(I) line, internal normalization with the N(I) line, and temperature correction are proposed and employed to satisfy the multi-line analysis method and yield the most accurate quantitative results. The calibration method for determining the organic oxygen content of coal is presented, with an accuracy of 1.15-1.37% and an average relative error of 19.39% being evaluated through an experiment performed on six anthracite coal samples. The relative measurement error distribution has also been studied. PMID- 18416907 TI - Barriers to eliminating waste in US health care. PMID- 18416908 TI - Are networks the answer to achieving integrated care? PMID- 18416909 TI - Do clinicians always maximize patient outcomes? A conjoint analysis of preferences for carotid artery testing. AB - OBJECTIVES: The value clinicians place on diagnostic information is subject to psychological influences and systematic biases, but there is limited evidence of how these factors may affect patients' health outcomes. We assessed the relative value attached by experienced clinicians to different diagnostic test characteristics and how their preferences relate to patient outcomes, focusing on strategies for testing symptomatic patients for carotid artery stenosis. METHODS: Using conjoint analysis, experienced neurologists and vascular surgeons ranked 10 diagnostic strategies defined in terms of four characteristics. Clinicians' preferences were analysed using an ordered probit model and compared with those obtained using a risk neutral expected value (EV) model developed to predict the consequences of each strategy as if the clinicians' sole goal were to optimize patient outcome. Results were tested for internal consistency and robustness to key model assumptions. RESULTS: Preferences for positive predictive value (PPV), relative to negative predictive value (NPV), elicited from the clinicians diverged substantially from those estimated by the EV model based on 5-year stroke-free survival (ratios of -0.8 and -32.8, respectively). Conversely, preferences for NPV, relative to test morbidity, from the two models matched closely. CONCLUSIONS: Clinicians attached substantially more importance to the PPV of carotid artery tests than would be justified by their impact on patient outcomes. Cognitive errors and attitudes to risk are likely to play an important role in explaining this finding. This study casts doubts on the validity of common assumptions made in the evaluation of health interventions, and in clinical and policy decisions. PMID- 18416910 TI - Patient choice in general practice: the implications of patient satisfaction surveys. AB - OBJECTIVES: To identify factors that explain patient satisfaction with general practice physicians and hence that may drive patients' choice of practice. METHODS: Logistic regression analysis of English National Health Service national patient survey data is used to identify the aspects of general practice care that are associated with high levels of overall satisfaction among patients. RESULTS: Confidence and trust in the doctor is the most important factor in explaining the variation in overall patient satisfaction (predicting 82% of satisfaction levels accurately). The seven variables relating to the relationship between patient and doctor have stronger explanatory power than other aspects of the general practitioner (GP) experience. The variables with the lowest overall predictive power are whether the patient was told how long they would have to wait in the surgery (72%), the length of time they had to wait after their appointment time (74%) and ability to get through to the surgery on the phone (74%). CONCLUSIONS: Patients value the quality of their relationship with their doctor more than the appearance of the surgery, accessibility of appointments and their experience in the waiting room. This suggests that, if current restrictions on choice of GP were removed, we would in theory expect a patient's choice to be driven by the quality of the doctor-patient relationship. Once a patient establishes a good relationship with a GP, however, we might expect them to be loyal and therefore unlikely to change practice unless the relationship with the doctor breaks down. Although relationship factors are important to the satisfaction of patients, it is not clear that they will lead large numbers of people to change their GP. PMID- 18416911 TI - Hospital clinicians' attitudes towards a statutory advocacy service for patients lacking mental capacity: implications for implementation. AB - OBJECTIVES: To examine attitudes to the Mental Capacity Act's new statutory Independent Mental Capacity Advocate (IMCA) service in England and Wales and consider the implications for its delivery. METHODS: Quantitative data describing all referrals to the seven pilot IMCA services (January 2006-March 2007) and qualitative data from semi-structured interviews with 18 doctors, 21 senior nurses and one discharge planning manager in four general hospitals in England. RESULTS: Of 127 hospital-based referrals to the seven pilot IMCA services, 29 (23%) were for patients facing serious medical treatments, 52% of whom were judged to lack decision-making capacity due to a learning disability; ninety eight (77%) were for patients facing a change of accommodation upon hospital discharge, 62% of whom were elderly and lacked capacity due to dementia. While aware of the potential benefits of the IMCA service, clinicians were generally negative about the contribution advocates could make to patients' medical care and thought they could only contribute usefully in a minority of ethically complicated decisions. In contrast, they were more positive about the involvement of advocates in hospital discharge decisions and hoped that they would improve current discharge practice. CONCLUSIONS: Clinicians held ambivalent attitudes towards the involvement of a statutory IMCA service in medical decisions, reflecting beliefs that the service was largely impractical and unnecessary given current procedures for making medical decisions in patients' 'best interests'. Conversely, clinicians were more likely to support advocacy in discharge decisions because they believed that non-medically qualified advocates could make a valuable contribution to decisions that were seen as predominantly social and where practice was frequently considered deficient. By holding these beliefs, clinicians are failing to have due regard for the IMCA service as a statutory measure for safeguarding patients' interests. PMID- 18416912 TI - Competing norms: Canadian rural family physicians' perceptions of clinical practice guidelines and shared decision-making. AB - OBJECTIVES: Implementation of clinical practice guidelines (CPGs) and shared decision-making are both advocated in primary care. Some authors argue that CPGs can enhance informed decisions by patients and physicians, while others warn that a standardized implementation of CPGs could hinder patients' involvement in decision-making. Our objective was to explore rural family physicians' perception of the interaction between clinical practice guidelines and shared decision making in medical practice. METHODS: A qualitative study using a semi-structured focus group interview: with 17 family physicians and residents, in a Canadian rural town. Interviews were audio-taped and transcribed verbatim. Thematic content analysis was performed and validated by the constant comparative method, member checking and group debriefing. RESULTS: Two distinct conceptions of how clinical practice guidelines should assist decision-making emerged. On the one hand, guidelines were seen as helping clinicians to make decisions on behalf of their patient about the best course of action. For interventions with uncertain benefit or that carried significant trade-off for patients, guidelines were seen as a tool that should inform decision-making between physicians and patients, providing them with details about risks, benefits, costs and alternative treatments. The pressure to apply guideline recommendations was perceived as a potential barrier to patient participation in decision-making. CONCLUSION: In circumstances where physicians judge patient participation in decision-making to be important, physicians perceive a tension between the need to respect patients' preferences and the pressure to apply guidelines. CPGs should include information that supports shared decision-making, besides their current focus on influencing prescription patterns, costs and health outcomes. PMID- 18416913 TI - Modelling the expected net benefits of interventions to reduce the burden of medication errors. AB - OBJECTIVES: The aim of this study is to estimate the potential costs and benefits of three key interventions (computerized physician order entry [CPOE], additional ward pharmacists and bar coding) to help prioritize research to reduce medication errors. METHODS: A generic model structure was developed to describe the incidence and impacts of medication errors in hospitals. The model follows pathways from medication error points at alternative stages of the medication pathway through to the outcomes of undetected errors. The model was populated from a systematic review of the medication errors literature combined with novel probabilistic calibration methods. Cost ranges were applied to the interventions, the treatment of preventable adverse drug events (pADEs), and the value of the health lost as a result of an ADE. RESULTS: The model predicts annual health service costs of between pound 0.3 million and pound 1 million for the treatment of pADEs in a 400-bed acute hospital in the UK. Including only health service costs, it is uncertain whether any of the three interventions will produce positive net benefits, particularly if high intervention costs are assumed. When the monetary value of lost health is included, all three interventions have a high probability of producing positive net benefits with a mean estimate of around pound 31.5 million for CPOE over a five-year time horizon. CONCLUSIONS: The results identify the potential cost-effectiveness of interventions aimed at medication errors, as well as identifying key drivers of cost-effectiveness that should be specifically addressed in the design of primary evaluations of medication error interventions. PMID- 18416914 TI - The quality of mixed methods studies in health services research. AB - OBJECTIVES: To assess the quality of mixed methods studies in health services research (HSR). METHODS: We identified 118 mixed methods studies funded by the Department of Health in England between 1994 and 2004, and obtained proposals and/or final reports for 75. We applied a set of quality questions to both the proposal and report of each study, addressing the success of the study, the mixed methods design, the individual qualitative and quantitative components, the integration between methods and the inferences drawn from completed studies. RESULTS: Most studies were completed successfully. Researchers mainly ignored the mixed methods design and described only the separate components of a study. There was a lack of justification for, and transparency of, the mixed methods design in both proposals and reports, and this had implications for making judgements about the quality of individual components in the context of the design used. There was also a lack of transparency of the individual methods in terms of clear exposition of data collection and analysis, and this was more a problem for the qualitative than the quantitative component: 42% (19/45) versus 18% (8/45) of proposals (p = 0.011). Judgements about integration could rarely be made due to the absence of an attempt at integration of data and findings from different components within a study. CONCLUSIONS: The HSR community could improve mixed methods studies by giving more consideration to describing and justifying the design, being transparent about the qualitative component, and attempting to integrate data and findings from the individual components. PMID- 18416915 TI - Can cheap generic statins achieve national cholesterol lowering targets? AB - OBJECTIVES: The Department of Health in England recommend that simvastatin and pravastatin should be prescribed in at least 69% of statin prescriptions in primary care on the assumption that these drugs (and at doses prescribed) are as effective as alternatives. We aimed to identify whether primary care trusts (PCTs) that used a high proportion of simvastatin and pravastatin performed as well on the Quality and Outcome Framework (QOF) targets related to cholesterol as those PCTs that used less. METHODS: QOF data were obtained for all PCTs for 2005 2006. National prescribing data for statins was analysed for the same time period. The square of the Pearson correlation was used to assess the association between the two. RESULTS: The average PCT values for the three QOF indicators for CHD, stroke and diabetes were 78% (range 66-88%), 72% (58-82%) and 79% (67-88%), respectively. The percentage use of simvastatin and pravastatin by PCTs varied from 18-84%, with a mean of 57%. There was no evidence of any association between the use of simvastatin and pravastatin as a percentage of all statin items and success in achieving the QOF targets. CONCLUSIONS: PCTs that had a high proportion of simvastatin and pravastatin use were just as successful achieving cholesterol targets for patients with coronary heart disease, diabetes and stroke as those that used more atorvastatin, rosuvastatin or fluvastatin. This supports the policy to use the less expensive generic statins. PMID- 18416916 TI - Evidence-based medicine and patient choice: the case of heart failure care. AB - OBJECTIVES: The implementation of evidence-based medicine and policies aimed at increasing user involvement in health care decisions are central planks of contemporary English health policy. Yet they are potentially in conflict. Our aim was to explore how clinicians working in the field of heart failure resolve this conflict. METHODS: Qualitative semi-structured interviews were carried out with health professionals who were currently caring for patients with heart failure, and observations were conducted at one dedicated heart failure clinic in northern England. RESULTS: While clinicians acknowledged that patients' ideas and preferences should be an important part of treatment decisions, the widespread acceptance of an evidence-based clinical protocol for heart failure among the clinic doctors significantly influenced the content and style of the consultation. CONCLUSION: Evidence-based medicine was used to buttress professional authority and seemed to provide an additional barrier to the adoption of patient-centred clinical practice. PMID- 18416917 TI - Decentralizing resource allocation: early experiences with district health boards in New Zealand. AB - OBJECTIVES: In New Zealand in 2001, a system of purchasing health services by a centralized purchasing agency was replaced by 21 district health boards (DHBs) which are responsible for both providing health services directly and for purchasing services from non-government providers. This paper describes the processes associated with the allocation of health resources in the decentralized system and considers the extent to which four of the government's stated objectives are likely to be achieved. METHODS: Two rounds of interviews with national stakeholders and senior DHB personnel plus case studies in five districts which included key informant interviews, observation at board meetings and document analysis. RESULTS: The re-structuring of the health sector in New Zealand appears to have simultaneously enhanced and inhibited the achievement of government objectives. Local decision-making has encouraged greater local responsiveness and new funding arrangements have allayed concerns about inter regional equity. The system is less commercially oriented than it was during the 1990s and collaboration between DHBs is improving. However, the combination of increased integration of purchasing and provision within DHBs and the focus on financial deficits in the early years appears to have inhibited the development of partnership relationships between DHBs and non-government providers, and of longer-term funding arrangements for high quality providers. Non-government providers perceive that DHBs have a tendency to favour their own providers when allocating contracts. CONCLUSIONS: Decentralized decision-making is starting to make some inroads towards achieving some of the government's objectives with respect to resource allocation and purchasing. PMID- 18416918 TI - World class commissioning: a health policy chimera? AB - The health reforms in England have entered a phase in which greater emphasis is being placed on market-like mechanisms. The ability of the commissioners of care to negotiate on equal terms with providers will be of critical importance in the emerging market. The government has set out plans to develop 'world class commissioning' and this essay reviews experience in Europe, New Zealand and the United States to understand what is involved in working towards this goal. The evidence reviewed shows that in no system is commissioning done consistently well and highlights the obstacles to the development of world class commissioning. The reasons for this centre on the complexity of health care and the inherent difficulty of commissioning health services in publicly financed systems. Commissioners will need to be able to access a range of expertise and are likely to incur significant expenditure in so doing. There are warning signs from other systems of health reforms that result in adversarial and legalistic approaches, and do not give sufficient attention to relational contracting. Even if world class commissioning is developed, it may fall short of its potential in the absence of other changes in the design of the reforms, such as autonomous providers and appropriate payment systems. In view of these challenges, a more promising alternative would be to develop competing integrated systems. PMID- 18416919 TI - Understanding determinants of health service use from a systems perspective. PMID- 18416922 TI - UK Health Services Research Network makes steady progress. PMID- 18416920 TI - Doctors, lies and the addiction bureaucracy. AB - Almost everything you know about heroin addiction is wrong. Not only is it wrong, but it is obviously wrong. Heroin is not highly addictive; withdrawal from it is not medically serious; addicts do not become criminals to feed their habit; addicts do not need any medical assistance to stop taking heroin; and contrary to received wisdom, heroin addiction most certainly is a moral or spiritual problem. A literary tradition dating back to De Quincey and Coleridge, and continuing up to the deeply sociopathic William Burroughs and beyond, has misled all Western societies for generations about the nature of heroin addiction. These writers' self-dramatizing and dishonest accounts of their own addiction have been accepted uncritically, and have been more influential by far in forming public attitudes than the whole of pharmacological science. As a result, a self-serving, self perpetuating and completely useless medical bureaucracy has been set up to deal with the problem. PMID- 18416923 TI - Is fast access to general practice all that should matter? A discrete choice experiment of patients' preferences. AB - OBJECTIVES: To determine the relative importance of factors that influence patient choice in the booking of general practice appointments for two health problems. METHODS: Two discrete choice experiments incorporated into a survey of general practice patients and qualitative methods to support survey development. RESULTS: An overall response of 94% (1052/1123) was achieved. Factors influencing the average respondent's choice of appointment, in order of importance, were: seeing a doctor of choice; booking at a convenient time of day; seeing any available doctor; and having an appointment sooner rather than later (acute, low worry condition). This finding was the same for an ongoing, high worry condition but in addition the duration of the appointment was also of (small) value. Patients traded off speed of access for more convenient appointment times (a willingness to wait an extra 2.5-3 days longer to get a convenient time slot for an acute low worry/ongoing, high worry condition, respectively). However, contrary to expectation, patients were willing to trade off speed of access for continuity of care (e.g. willingness to wait five days longer to see the doctor of their choice for an acute, low worry condition). Preferences varied by a person's gender, work and carer status. CONCLUSIONS: Patients hold strong preferences for the way general practice appointment systems are managed. Contrary to current policy on improving access to primary care patients value a more complex mix of factors than fast access at all costs. It is important that policy-makers and practices take note of these preferences. PMID- 18416924 TI - Legitimacy of hospital reconfiguration: the controversial downsizing of Kidderminster hospital. AB - OBJECTIVES: This paper examines the contested organizational legitimacy of hospital reconfiguration, which continues to be a central issue in health care management. METHODS: A qualitative study which focuses on the controversial downsizing of Kidderminster Hospital, a highly publicized landmark case of district general hospital closure. Rhetorical strategies are analysed to examine how legitimacy was constructed by stakeholder groups and how these strategies were used to support or resist change. RESULTS: Stakeholders promoting change legitimized re-organization pragmatically and morally arguing the need for centralization as a rational necessity. Stakeholders resisting change argued for cognitive and moral legitimacy in current service arrangements, contrasting local versus regionalized aspects of safety and provision. Groups managed to talk past each other, failing to establish a dialogue, which led to significant conflict and political upheaval. CONCLUSIONS: Stakeholders value hospitals in different ways and argue for diverse accounts of legitimacy. Broader discourses of medical science and democratic participation were drawn into rhetorical texts concerning regionalization to render them more powerful. PMID- 18416925 TI - Characteristics of the emergency and urgent care system important to patients: a qualitative study. AB - OBJECTIVES: To explore patients' views and experiences of the emergency and urgent care system to inform the development of a questionnaire for routine assessment of the system's performance from the patient perspective. METHODS: Qualitative research with people who had recently used the system: 47 people in eight focus groups and 13 individual interviews. RESULTS: Recurrent themes included characteristics of the system which are rarely addressed in service specific questionnaires, in particular, confusion over the most appropriate service to use for particular health problems, coordination between services and informational continuity across services. Other characteristics were identified which, although commonly included in service-specific questionnaires, could have system-level consequences. These included communication between health professionals and patients, and ease of access to services. For example, patients' perception of poor communication with one service could increase their subsequent use of other services in the system. Proactive behaviour from health professionals was an important system characteristic because it could allay patient anxiety by making patients feel that their concerns were being taken seriously and that staff could sort out problems such as feeling 'stuck in' or 'bounced around' the system. 'Candidacy', whereby eligibility for health care is jointly negotiated between the user and the service provider, was evident as an issue for patients across the social spectrum when seeking help urgently. CONCLUSIONS: Questionnaires designed to assess patients' views and experiences of emergency and urgent care should address system-level as well as service-specific issues in order to address the full range of patient concerns. PMID- 18416926 TI - Equity and service innovation: the implementation of a bibliotherapy scheme in Wales. AB - OBJECTIVE: Book Prescription Wales (BPW) is a pilot bibiliotherapy scheme launched in July 2005 as a primary care treatment option for people with mild to moderate mental health problems. In an innovative model, patients are prescribed self-help books from a list, to borrow from local libraries. Our objective was to evaluate its implementation, focusing on the issue of equity of service delivery. METHODS: Data were gathered from Welsh Assembly Government concerning project set up and borrowing rates. Mailed questionnaires were completed by 21/22 (95.4%) Local Health Boards and 44/64 (68.8%) Community Mental Health Teams. In addition, 327 out of 497 (66%) primary care practices were surveyed by telephone, 20 prescribers took part in in-depth telephone interviews and three focus groups were conducted with library staff. RESULTS: From July 2005-March 2006, books were borrowed 15,236 times. There was a 10-fold variation in borrowing rates across local authorities (1.07 to 10.18 loans/1000 people). The priority which Local Health Board staff reported giving to the scheme varied. Uptake among prescribers was mixed: in 35% of general practices (n = 116) no-one participated. Prescribers reported different ways of using the bibliotherapy scheme. Library staff reported issues of patchy uptake. CONCLUSION: Variation in usage of bibliotherapy raises questions about equity; it is unlikely to reflect the distribution of people who could potentially benefit. Factors influencing variation existed all along the implementation chain. It is not always possibly to separate demand-side and supply-side factors when considering equity and service innovation in health care. PMID- 18416927 TI - Part of a global workforce: migration of British-trained pharmacists. AB - OBJECTIVES: Many countries, including the UK, have identified a shortage of pharmacists, partly due to emigration. This study was undertaken to examine the extent and nature of migration taking place among British-qualified pharmacists. METHODS: Mixed methods, including secondary analysis of quantitative data, qualitative research and a large self-completion survey of all British-registered pharmacists with an overseas address. RESULTS: Almost 11% of British-registered pharmacists reside overseas. Nearly three-quarters are British-trained and most are UK nationals. The US, Canada and Australia are the main destinations. The majority work as pharmacists in health services, but sizeable proportions are either retired, not working for other reasons or work in industry. Those who emigrate include those returning home, moving for career opportunities, for lifestyle reasons or as a 'spouse trailer'. For many the move abroad is a permanent one. CONCLUSIONS: Great Britain is both a source and destination country for migrating pharmacists. Emigration currently exceeds immigration. Pharmacists are not migrating to developing countries, so the profession may want to consider ways of contributing to the health care systems in developing countries which are the source of some of the immigrant pharmacists to Great Britain. PMID- 18416928 TI - The increasing burden of alcoholic liver disease on United Kingdom critical care units: secondary analysis of a high quality clinical database. AB - OBJECTIVES: To investigate the effect of increasing alcohol consumption on the number of admissions to adult, general critical/intensive care units (ICUs) in England and Wales with alcoholic liver disease, their case mix, mortality, and impact on critical care and hospital activity by extrapolating from admissions to ICUs over the last 10 years. METHODS: Secondary analysis of a high quality clinical database from a national clinical audit using data from 385,429 admissions to174 ICUs in England and Wales between December 1995 and July 2005, of which 4219 (1.1%) had alcoholic liver disease. The extrapolated total number of admissions with alcoholic liver disease and total number of ICU bed-days occupied were calculated. Changes over time in the case mix (age, sex and APACHE II and ICNARC risk prediction models), mortality at ultimate discharge from acute hospital, and length of stay in ICU and in hospital were explored. RESULTS: The percentage of ICU admissions with alcoholic liver disease increased from 0.65% in 1996 to 1.35% in 2005, but the case mix remained similar. Mortality decreased and length of stay increased over this period. The extrapolated total number of admissions to all 229 adult, general critical care units in England and Wales increased from 550 in 1996 to 1513 in 2005, and the extrapolated total number of bed-days occupied by these admissions increased from around 3100 to over 10,000. CONCLUSIONS: Admissions to ICUs in England and Wales with alcoholic liver disease tripled over the 10-year period from 1996 to 2005. The continuing increase in alcohol consumption means that this trend is likely to continue. PMID- 18416929 TI - Career intentions of pharmacy students. AB - OBJECTIVES: In light of pharmacy workforce shortages in Great Britain, the profession's regulatory body commissioned a programme of longitudinal work to explore pharmacy career decision-making in relation to influences on career choice and intended career paths. Our objective was to gather data on career intentions that could be used to produce robust predictions about pharmacist supply. METHODS: Two annual surveys conducted with the same cohort of pharmacy students in 2005 and 2006. The questionnaires sought to clarify influences on respondents' career intentions and their early career plans. RESULTS: Only two thirds of respondents intended going straight into British pharmacy practice after training, with fewer white men (57%) than ethnic minority men (71%) intending to go straight into practice. Preferences for early careers reflected existing occupational segregation, with 41% of white females hoping to work in hospital pharmacy and a similar proportion of ethnic minority men (40%) hoping to work for a large multiple community pharmacy after training. CONCLUSIONS: A sizeable proportion of pharmacy students do not intend entering the profession for which they have trained, a proportion which is much larger than estimated by other studies. This has significant implications for workforce planning. Existing gender and ethnic segregation in the profession may have occurred as a result of personal choice rather than being a function of constraints operating within the pharmacy labour market. PMID- 18416930 TI - The local adaptation of national recommendations for preventing early-onset neonatal Group B Streptococcal disease in UK maternity units. AB - OBJECTIVE: To evaluate how UK maternity unit protocols conformed to national recommendations for preventing early-onset neonatal Group B Streptococcal (GBS) disease. METHODS: In December 2005, all UK obstetric maternity units were contacted and asked to provide a copy of their protocol on preventing GBS disease. Information was extracted on the protocol's recommendations, its development date and the evidence cited. The protocol's recommendations were then compared against the recommendations in the Royal College of Obstetricians and Gynaecologists (RCOG) guideline. RESULTS: Protocols were obtained for 171 of the 227 units (75%), of which 120 were developed after the guideline has been published. There were 134 protocols (78%) that followed the RCOG prevention strategy, recommending a risk-based approach to selecting women for intrapartum antibiotic prophylaxis (IAP). However, the sets of risk factors named as indications for IAP differed between the protocols and only 34 of these 134 protocols were entirely consistent with the guideline. The 37 protocols (22%) that did not follow the RCOG prevention strategy recommended IAP for some risk factors but only if a bacteriological test was also GBS positive. CONCLUSIONS: There are considerable differences in the GBS protocols used in maternity units in the UK despite the availability of a national guideline. Consequently, some high-risk women may not receive IAP while some women without risk factors are treated needlessly. While local adaptation may be for legitimate reasons, the processes used in some units seem to require improvement. PMID- 18416931 TI - Learning from other countries: an on-call facility for health care policy. AB - Recognizing that robust information on health systems in other countries can provide valuable lessons for the English National Health Service, the Department of Health commissioned an academic team to provide an 'On-call Facility for International Healthcare Comparisons' in 2005. This paper describes the work of this novel approach to informing policy and reviews the experience of the first two years. It illustrates the well-documented challenges of comparative analysis of health systems. One important issue is understanding the health system context so as to interpret phenomena and draw appropriate policy conclusions. Other challenges include the potential tension between academic interest and rigour, and the need for timely analysis to inform the Department of Health's rapidly changing policy agenda. The diversity and nature of topics covered, as well as the rapid turn-around time have meant that the Facility has had to balance rigour and timeliness carefully to ensure the value and relevance of reports. A strong research base linked with an international network of country experts promotes the provision of high quality analyses at relatively low costs. However, such an arrangement can only be sustained if it provides scope for additional primary research. A formal evaluation of the influence on health care policy-making in England is not yet available. Such knowledge will be of crucial importance for the development of similar resources elsewhere. PMID- 18416932 TI - Prediction of human pharmacokinetics-biliary and intestinal clearance and enterohepatic circulation. AB - The main objective was to evaluate and propose methods for predicting biliary clearance (CL(bile)) and enterohepatic circulation (EHC) of intact drugs in man. Another aim was to evaluate to role of intestinal drug secretion and propose a method for prediction of intestinal secretion CL (CL(i)). Animal data poorly predict the CL and CL(bile) of biliary excreted drugs, and the suggested molecular weight threshold for bile excretion as the dominant elimination route does not seem to hold. Active transport, low metabolic intrinsic CL (CL(int)) and, as an approximation, permeability (P(e)) less than that of metoprolol is required for substantial CL(bile) to occur. The typical EHC plasma concentration vs time profile (multiple peaks) is demonstrated for many low metabolic CL(int) compounds with efflux and moderate to high intestinal P(e) and fraction absorbed. Physiologically-based in-vitro to in-vivo (PB-IVIV) methodology with in-vitro intrinsic CL(bile)-data obtained with sandwich-cultured human hepatocytes has generated 2- and 5-fold underpredictions for two compounds with intermediate to high CL(bile). This is despite not considering the unbound fraction. Possible explanations include low transporter activity and diffusion limitations in the in vitro experiments. Intestinal reabsorption and EHC were also neglected in these predictions and in-vivo CL(bile) estimations. The sandwich model and these reference data are still very useful. Consideration of an empirical scaling factor and a newly developed approach that accounts for intestinal reabsorption and EHC could potentially lead to improved PB-IVIV predictions of CL(bile). Apparently, no attempts have been made to predict CL(i). Elimination via the intestinal route does not appear to be of great importance for the few compounds with available data, but could be equally as important as bile excretion. Net secretion in-vitro P(e) and newly estimated in-vivo intrinsic CL(i) data for digoxin and rosuvastatin could be useful for approximation of CL(i) of other compounds. PMID- 18416933 TI - Pharmaceutical and pharmacological importance of peptide transporters. AB - Peptide transport is currently a prominent topic in membrane research. The transport proteins involved are under intense investigation because of their physiological importance in protein absorption and also because peptide transporters are possible vehicles for drug delivery. Moreover, in many tissues peptide carriers transduce peptidic signals across membranes that are relevant in information processing. The focus of this review is on the pharmaceutical relevance of the human peptide transporters PEPT1 and PEPT2. In addition to their physiological substrates, both carriers transport many beta-lactam antibiotics, valaciclovir and other drugs and prodrugs because of their sterical resemblance to di- and tripeptides. The primary structure, tissue distribution and substrate specificity of PEPT1 and PEPT2 have been well characterized. However, there is a dearth of knowledge on the substrate binding sites and the three-dimensional structure of these proteins. Until this pivotal information becomes available by X-ray crystallography, the development of new drug substrates relies on classical transport studies combined with molecular modelling. In more than thirty years of research, data on the interaction of well over 700 di- and tripeptides, amino acid and peptide derivatives, drugs and prodrugs with peptide transporters have been gathered. The aim of this review is to put the reports on peptide transporter-mediated drug uptake into perspective. We also review the current knowledge on pharmacogenomics and clinical relevance of human peptide transporters. Finally, the reader's attention is drawn to other known or proposed human peptide-transporting proteins. PMID- 18416934 TI - Skin-compatible lecithin drug delivery systems for fluconazole: effect of phosphatidylethanolamine and oleic acid on skin permeation. AB - The purpose of the present study was to evaluate skin-compatible drug delivery systems for fluconazole. Pseudoternary phase diagrams were constructed, composed of different soybean lecithins/oil/isopropanol and water. The role of the various lecithin compositions was expressed in the different resulting isotropic areas. Based on these phase diagrams, two systems were chosen as drug delivery systems for fluconazole. The influence of phosphatidylethanolamine and of the oil component on the skin permeation of fluconazole was investigated. The more phosphatidylethanolamine, the greater was the fluconazole skin permeation, independent of the hydrophilicity of the system. The influence of oleic acid and isopropylmyristate as the oil component was compared and a greater penetration enhancing effect was found for the microemulsion containing oleic acid. PMID- 18416935 TI - Effect of molecular weight and end-group nature on the solubility of ethylene oxide oligomers in 2H, 3H-decafluoropentane and its fully fluorinated analogue, perfluoropentane. AB - Fluorinated liquids possess high chemical and physical stability, are tolerated by the human body and, therefore, show great promise in biomedical fields; however, they require extensive formulation. Phase diagrams are reported here for a series of ethylene oxide oligomeric additives in 2H,3H-perfluoropentane (HPFP), a non-chlorofluorocarbon fluorinated liquid regarded as a model propellant for pressurized metered-dose inhalers. Over a wide range of temperatures and concentrations, dihydroxyl end-capped poly(ethylene glycols) (PEGs) exhibited a lower critical solution temperature (LCST) that was strongly molecular weight dependent. In contrast, monomethyl (and thus monohydroxy) and dimethyl end-capped poly(ethylene oxides) were fully miscible with HPFP over the same temperature and concentration ranges, suggesting that the phase behaviour was dominated by end group/solvent interactions. By systematically substituting HPFP for the fully fluorinated analogue perfluoropentane, the ability of these end-groups to interact with the solvent was perturbed and LCST-type behaviour was induced in the previously fully miscible monomethyl and dimethyl end-capped PEGs. Concomitantly, with increasing perfluoropentane content, the LCST of the dihydroxyl end-capped PEGs was driven to lower temperatures. Therefore, the phase behaviour of these systems may be controlled by 'tuning' the end-group structure of the ethylene oxide oligomers, and varying the hydrogen bonding capabilities of the fluorinated solvents. PMID- 18416936 TI - Nebulisation of corticosteroid suspensions and solutions with a beta(2) agonist. AB - The aim of this study was to determine the output of salbutamol nebulised in combination with either flunisolide or beclometasone dipropionate (BDP) from two different nebulisers under simulated breathing conditions. The BimboNeb and Nebula nebulisers were used to nebulise 3.0 mL of the two drug mixtures (salbutamol, 5000 microg plus either flunisolide, 600 microg, or BDP, 800 microg). Particle size was determined by inertial impaction. Total outputs of all drugs from both nebulisers were measured using a sinus flow pump under simulated paediatric and adult breathing patterns. The mass median aerodynamic diameter (MMAD) of BDP particles from the mixture was 6.34 mum using the BimboNeb and 5.34 mum using the Nebula. Values for salbutamol in this mixture were 3.93 and 3.32 microm, respectively. The MMAD of flunisolide particles from the BimboNeb and Nebula were 3.74 and 3.65 microm, respectively, while for salbutamol were 3.79 and 3.74 microm, respectively. With the simulated adult breathing pattern, all drug outputs from both mixtures were greater from the BimboNeb than from the Nebula after 5 and 10 min' nebulisation. Drug delivery from the BimboNeb, but not the Nebula, was affected by the simulated breathing pattern. Outputs with the BimboNeb were lower with the paediatric breathing pattern than with the adult pattern. In the majority of cases, nebulising for 10 min produced significantly greater drug output than after 5 min. For the Nebula, outputs were generally similar at 5 and 10 min, irrespective of the breathing pattern. These results highlight the need to assess the amount of aerosolised drug available when drugs are combined, when different nebulisers are used and when they are used with patients of different ages. PMID- 18416937 TI - Optimizing delivery of flurbiprofen to the colon using a targeted prodrug approach. AB - The carboxylic group responsible for the gastric side-effects of the propionic acid derivative, flurbiprofen, was masked temporarily to overcome these side effects and to accomplish colon-specific delivery of the drug. An amide prodrug (FLU-GLY) was synthesized by coupling flurbiprofen with L-glycine. Confirmation and characterization of the structure of the synthesized prodrug included elemental analysis, Fourier transform (FT)-IR, FT-NMR, mass (FAB) spectroscopy, and determinations of R(f), R(t) and R(M) values, respectively. Aqueous solubility and lipophilicity (logP) value were determined at pH 1.2, 4.0, 6.8 and 7.4. In-vitro reversion of FLU-GLY to flurbiprofen was measured at different pHs and in a simulated colonic environment. Acute toxicity and ulceration potential were evaluated in-vivo in albino rats. Pre-formulation studies showed increased hydrophilicity but a non-significant increase in lipophilicity of the prodrug. In vitro reversion studies suggested that the prodrug remained intact until colonic pH was attained, when the colonic microfloral enzymes (amidase) hydrolysed the FLU-GLY amide linkage, releasing the free drug. In-vivo evaluation indicated that the prodrug was much less toxic and had less ulcerogenic activity than the parent drug. Selective delivery of drugs to the colon can be useful in terms of reducing the dose administered and reducing undesirable side-effects. PMID- 18416938 TI - In-vivo evaluation in rats of colon-specific microspheres containing 5 fluorouracil. AB - The aims of this investigation were to determine the distribution in the gastrointestinal (GI) tract of Eudragit S-100 encapsulated colon-specific sodium alginate microspheres containing 5-fluorouracil (5-FU) in rats, and to perform pharmacokinetic and pharmacodynamic studies. Comparisons were with a control immediate-release (IR) formulation of 5-FU. 5-FU was distributed predominantly in the upper GI tract from the IR formulation but was distributed primarily to the lower part of the GI tract from the microsphere formulation. No drug was released in the stomach and intestinal regions from the colon-specific microspheres. Significantly, a high concentration of the active drug was achieved in colonic tissues from the colon-specific microspheres (P < 0.001), which was higher than the IC50 required to halt the growth of and/or kill colon cancer cells. Colon cancer was induced in rats by subcutaneous injection of 1,2-dimethylhydrazine (40 mg kg (-1)) for 10 weeks. The tumours induced were non-invasive adenocarcinomas and were in Duke's stage A. The 5-FU formulations were administered for 4 weeks after tumour induction. Non-significant reductions in tumour volume and multiplicity were observed in animals given the colon-specific microspheres. Enhanced levels of liver enzymes (SGOT, SGPT and alkaline phosphatase) were found in animals given the IR formulation of 5-FU, and values differed significantly (P < 0.001) from those in animals treated with the colon-specific microspheres. Elevated levels of serum albumin and creatinine, and leucocytopenia and thrombocytopenia were observed in the animals given the IR formulation. In summary, Eudragit S-100 coated alginate microspheres delivered 5-FU to colonic tissues, with reduced systemic side-effects. A long-term dosing study is required to ascertain the therapeutic benefits. PMID- 18416939 TI - Effects of morin on the pharmacokinetics of nicardipine after oral and intravenous administration of nicardipine in rats. AB - This study investigated the effects of orally administered morin, an inhibitor of cytochrome P450 3A (CYP3A) and P-glycoprotein (P-gp), on the pharmacokinetics of orally and intravenously administered nicardipine in rats. Nicardipine is reportedly a substrate for CYP3A4 and P-gp. Nicardipine was administered orally (12 mgkg(-1)) with or without orally administered morin (1.5, 7.5 and 15 mgkg( 1)), and intravenously (4 mgkg(-1)) with or without orally administered morin (7.5 and 15 mgkg(-1)). In the presence of morin, the pharmacokinetic parameters of nicardipine were significantly altered in the oral group but not in the intravenous group, suggesting that CYP3A-mediated metabolism of nicardipine in the liver is not significantly inhibited by morin. The presence of 7.5 and 15 mgkg(-1) of morin significantly increased (P < 0.01, 67.8-112%) the area under the plasma concentration-time curve and the peak plasma concentration (P < 0.01, 53.5-93.1%) of orally administered nicardipine. The presence of 7.5 and 15 mgkg( 1) of morin significantly decreased (P < 0.01, 40.4-52.8%) the total body clearance of orally administered nicardipine compared with the control group. The enhanced oral bioavailability of nicardipine suggests that intestinal-mediated CYP3A4 metabolism and P-gp-mediated efflux of nicardipine are inhibited by morin. Based on these results, concomitant use of morin or morin-containing dietary supplements with nicardipine may require close monitoring for potential drug interactions. PMID- 18416940 TI - ABCC2 haplotype is not associated with drug-resistant epilepsy. AB - Several studies have investigated the association between the ABCB1 polymorphism and drug-resistant epilepsy. However, the effect of ABCC2 polymorphisms on anti epileptic drug (AED) responsiveness remains unknown. The ABCC2 polymorphisms have been genotyped in 279 Japanese epileptic patients treated with AEDs. The association between the AED responsiveness and the polymorphisms was estimated by a haplotype-based analysis. No genotype or haplotype was associated with drug resistant epilepsy. On the other hand, the delGCGC haplotype at G-1774delG, C 24T, G1249A and C3972T was over represented among the epileptic patients with a complication of mental retardation in comparison with those without (32.4% vs 22.0%; P=0.009); and the G-1774delG allele was also associated with mental retardation (P=0.03). No association between the ABCC2 genotypes or haplotypes, and the responsiveness of AEDs was observed, although this finding was inconclusive because of the small sample size. PMID- 18416941 TI - Synthesis, pharmacological activity and nitric oxide generation by nitrate derivatives of theophylline. AB - Nitrates of theophylline derivatives - potential nitric oxide (NO) donors - were synthesized by esterification of 7-hydroxyalkyl theophylline derivatives with fuming nitric acid. The nitrates obtained were tested in-vitro in reactions with sulfydryl compounds at appropriately adjusted pH and temperature. Under the applied conditions, the synthesized compounds underwent decomposition to release NO, quantified using a polarographic method using a selective isolated (ISO-NO) sensor. The effects of dyphylline and proxyphylline and their new synthesized nitrates on arterial blood pressure (BP) were measured in spontaneously hypertensive (SH) rats. BP was measured in conscious SH rats using the tail-cuff method. Both short- and long-term administration of the xanthines tested significantly decreased systolic, diastolic and mean BP. The hypotensive effect of a single dose of nitrate dyphylline on mean BP was greater than that of the parent compound (P = 0.000012; P=0.000472 at 30 and 60 min post-dose, respectively), whereas proxyphylline and its nitrate derivative had similar activity. In rats treated with the tested compounds for 9 days twice daily, the decrease in BP persisted for at least 16 h after the last dose. Proxyphylline produced the most marked decrease in diastolic and mean BP. Among the xanthines examined, proxyphylline nitrate had the strongest hypotensive effect when administered in a single dose to animals pretreated with the same compound for 9 days. These results indicate that insertion of a nitrate group weakly modifies the hypotensive action of the studied xanthines in SH rats. PMID- 18416942 TI - The ability of selected pyridinium salts to increase the cytotoxic activity of vincristine but not doxorubicin towards sensitive and multidrug resistant promyelocytic leukaemia HL60 cells. AB - The aim of this study was to examine the effect of selected pyridinium salts, 1 methyl-3-nitropyridine chloride (MNP(+)Cl(-)) and 3,3,6,6,10-pentamethyl-3,4,6,7 tetrahydro-[1,8(2H,5H)-dion]acridine chloride (MDION(+)Cl(-)), on the activity of doxorubicin (DOX) and vincristine (VINC) towards human promyelocytic leukaemia HL60 cells as well as its multidrug resistant (MDR) sublines exhibiting two different phenotypes of MDR related to the overexpression of P-glycoprotein (HL60/VINC) or MRP1 (HL60/DOX). MNP and MDION salts were much less cytotoxic themselves (about 100-fold and 2000-fold compared with DOX and VINC, respectively) against HL60 cells but, in contrast to DOX and VINC, they conserved an important cytotoxic activity towards resistant HL60/VINC and HL60/DOX cells (resistance factor, RF = 2-4.5). It was shown that MNP(+)Cl(-) and MDION(+)Cl(-) increased the cytotoxicity of non-bioreductive antitumour agent VINC towards human promyelocytic leukaemia HL60 cells and its resistant sublines HL60/VINC and HL60/DOX. However, in the case of DOX the decrease in its cytotoxic activity towards all studied cell lines was observed in the presence of MNP(+)Cl(-) and MDION(+)Cl(-). Presented data suggest that the bioreductive drug DOX, in contrast to VINC, could compete with pyridinium salts (MNP(+)Cl(-) and MDION(+)Cl(-)) for NADPH-dependent oxidoreductases and for undergoing cellular reductive activation. This could explain the inefficiency of these salts to increase the cytotoxic activity of DOX against examined leukaemic HL60 cell line and its MDR sublines, HL60/VINC and HL60/DOX. PMID- 18416943 TI - Omeprazole promotes gastric epithelial cell migration. AB - Proton pump inhibitors (PPIs) are effective at preventing non-steroidal anti inflammatory drug (NSAID)-induced gastric ulcers. They are also superior to histamine H(2)-receptor antagonists and misoprostol in treating NSAID-induced gastric ulcer healing. This study explored whether omeprazole, a PPI, can modulate ulcer healing through epithelial cell proliferation and/or cell migration using a rat normal gastric epithelial cell line (RGM-1). Flow cytometry was used to determine cell proliferation and an artificial wound model was used to measure cell migration. Western blot analysis was performed to evaluate the possible mechanisms of action. Omeprazole treatment (10(-8), 10(-6) and 10(-4)M) for 12 and 24 h did not promote cell proliferation. However, similar doses of the drug (10(-6) and 10(-4)M) incubated for 24-48 h significantly promoted the basal cell migration of gastric epithelial cells. Further, the higher concentration of omeprazole (10(-4)M) reversed the inhibitory action of indometacin (10(-5)) on cell migration. Western blot results showed that omeprazole did not increase cyclooxygenase-2 expression and did not activate signal transduction pathways, including extracellular signal-regulated kinase (ERK1/ERK2), P38 mitogenic activated protein kinase, and phosphatidyl inositol 3-kinase. The results suggest that omeprazole is beneficial in basal ulcer healing and it reversed the adverse action of indometacin on ulcer repair under acid-independent conditions. These actions are likely to be mediated through the promotion of gastric epithelial cell migration but not cell proliferation. PMID- 18416944 TI - Effects of glycyrrhetinic acid and liquorice extract on cell proliferation and prostate-specific antigen secretion in LNCaP prostate cancer cells. AB - Glycyrrhetinic acid (GA) is the active metabolite of glycyrrhizic acid, one of the components of liquorice extract. It has been shown to possess anti inflammatory activity and to inhibit hepatic tumour growth. In this preliminary study, we have shown that GA could significantly reduce the rate of proliferation of LNCaP androgen dependent prostate cancer cells, whereas it had no effect on proliferation of PC3 and DU145 androgen-independent prostate cancer cells. Additionally, GA could significantly reduce the production of prostate-specific antigen by LNCaP cells maintained in-vitro. This study provides a sound platform for further investigation. PMID- 18416945 TI - The pharmacokinetics of antofloxacin in renally impaired rats. AB - Our aim was to investigate whether renal impairment induced by cisplatin altered the pharmacokinetics of antofloxacin. Antofloxacin (7.5 mg kg(-1), i.v.) was given to normal or renally impaired rats (induced by cisplatin). Concentrations of antofloxacin in plasma and urine were measured using HPLC. Pharmacokinetic parameters were estimated. The plasma concentrations of antofloxacin in the renally impaired rats were significantly higher than those in the normal rats, accompanied by significant increase of the area under the plasma concentration time curve (AUC) (968.78+/-259.39 microg min mL(-1) versus 509.84+/-46.19 microg min mL(-1) in normal rats P < 0.05). The system clearance (CL) and renal clearance (CL(R)) of antofloxacin decreased from 12.66+/-1.15 mL kg(-1) min(-1) and 3.21+/-1.80 mL kg(-1) min(-1) in normal rats, to 6.63+/-2.82 mL kg(-1) min( 1) and 0.31+/-0.15 mL kg(-1)min(-1), respectively. No differences between two treatments in half-life and mean residence time were found. We concluded that renal impairment induced by cisplatin significantly altered the pharma-cokinetics of antofloxacin and resulted in decrease of the renal elimination. PMID- 18416946 TI - The English national screening programme for sight-threatening diabetic retinopathy. AB - OBJECTIVES: The main objective of the national screening programme is to reduce the risk of sight loss among people with diabetes due to diabetic retinopathy (DR). METHODS: Offering two-field mydriatic digital photographic screening to all people with diabetes in England over the age of 12 years. STAGE OF DEVELOPMENT: The programme is in its infancy, receiving the first year's annual reports from approximately 96 screening programmes, each of which have developed to offer screening to a minimum number of 12,000 people with diabetes, which would cover a population of 350,000 people with 3.4% diabetes prevalence. The national programme has commenced the External quality assurance (QA) programme in order to achieve and sustain the highest possible standards. POTENTIAL BENEFITS: England has a population of two million people with diabetes over the age of 12 and it is believed that there is a prevalence of blindness of 4200 and an annual incidence of blindness of 1280 people with diabetes. This programme has the potential to reduce the prevalence of blindness in England from 4200 people to 1000 people and a conservative estimate of reducing the annual incidence of DR blindness by one third would save 427 people per annum from blindness. These figures are based on the UK certification of blindness but if World Health Organization (WHO) definitions are used the prevalence, incidence and potential reductions in blindness are much greater. PMID- 18416947 TI - The implementation of revised guidelines and the performance of a screening programme for congenital hypothyroidism. AB - OBJECTIVES: To see whether revised screening standards and health-professional training are associated with changes in the performance of a neonatal screening programme for congenital hypothyroidism (CHT). METHODS: Screening data from the regional screening service in Durham and Newcastle, which covers north-east England and North Cumbria. SETTING: We assessed the timing of the different stages of the screening process leading up to the introduction of the revised guidelines between April 2004 and March 2005 (year 1) and afterwards between April 2005 and March 2006 (year 2) in all babies notified as having CHT. We also assessed the interval between sampling and specimen arrival in the laboratory at the beginning and end of year 2 in all babies screened. RESULTS: Twenty-three babies tested positive or borderline in year 1 and 18 babies in year 2. There was reduced variability in the overall time from birth to notification in year 2 versus year 1 (P = 0.001). This reduction was a consequence of a reduced interval between sample collection and arrival in the laboratory (P = 0.047) and for the laboratory to notify the positive test result (P = 0.003). There was a reduction in the mean time from sampling to receipt by the laboratory in the 2997 babies screened in the final month compared with the 2498 babies screened in the first month of year 2 (P = 0.01). CONCLUSION: There was an improvement in neonatal screening programme performance around the time that revised neonatal screening guidelines were introduced. This highlights the importance of ongoing education and training for those involved in screening programmes. PMID- 18416948 TI - Implementation of the newborn screening programme for sickle cell disease in England: results for 2003-2005. AB - OBJECTIVES: This paper reports early screening results from the newborn sickle cell disease screening programme recently implemented in England. SETTING: England. Screening is offered at 5-8 days of age as part of the existing bloodspot test and offered to all babies irrespective of ethnicity. METHODS: The laboratory methods recommended are high performance liquid chromatography (HPLC) and iso-electric focusing (IEF). Two methods of analysis must be applied to all screen positive results. The conditions screened for are:- Sickle cell anaemia (Hb SS), Hb SC disease, Hb S/beta-thalassaemia, Hb S/D(Punjab), Hb S/O(Arab), Hb S/HPFH. Carriers identified for the common haemoglobin variants are reported to parents and follow-up counselling is offered. A bespoke laboratory quality assurance programme has been established which has defined standards of satisfactory performance. RESULTS: Provisional figures from the first seven months of screening (up to March 2004) 108,255 infants were screened gave a screen positive rate of 1:900 for these high prevalence areas and a carrier rate of 2.7%. Figures for 2004-2005 show about 250 significant screen positive results for sickle cell disorders and about 6,500 carriers were identified. The birth prevalence for screen positive results from 2004-05 is 1:1500. We estimate that when there is countrywide data, the national birth prevalence will be about 1:2000-1:2,500. CONCLUSION: The results from the national newborn sickle cell screening programme in England-show that the sickle cell disorders are as common as cystic fibrosis (CF) in England, although the distribution of cases is concentrated in London and other urban areas. The findings and approach to implementation adopted in England may be of interest to other Western European countries with increasing rates of sickle cell disease who are considering such programmes and also to other developed countries. PMID- 18416949 TI - The effect of an organized, nationwide breast cancer screening programme on non organized mammography activities. AB - OBJECTIVES: To analyse the effect of an organized, nationwide breast cancer screening programme on non-organized mammography activities in Hungary. SETTING: The nationwide dataset of the Hungarian National Health Insurance Fund Administration covering the years 2000-2005. METHODS: Data derived from the nationwide database of the Hungarian National Health Insurance Fund Administration. The study includes all women undergoing mammography before (2000 2001) and after (2002-2003/2004-2005) the introduction of organized screening. RESULTS: The number of women having non-organized (opportunistic/diagnostic) mammograms was around 250,000 in 2000-2001, but increased to 350,000 in 2005. In the age group 45-64 years in 2000-2001, only 27.4% of all women undergoing mammography were examined within locally-organized programmes. After the introduction of the nationwide programme, this percentage increased to 61.0% in 2002-2003, and 56.3% in 2004-2005. After the introduction of the nationwide organized programme (2002-2003), the proportion of organized screening mammographies remained among the highest in county Hajdu-Bihar (78.4%) and Zala (88.3%) and increased significantly in county Vas (87.7%). CONCLUSION: The introduction of an organized nationwide screening programme in Hungary resulted in increases in the number of screening mammographies, and also of non-organized mammographies. Although the ratio of organized screening versus non-organized mammography changed in favour of screening mammographies, there are large within country differences between counties. PMID- 18416950 TI - Estimating the cumulative risk of a false-positive under a regimen involving various types of cancer screening tests. AB - OBJECTIVES: When evaluating screening for the early detection of cancer, it is important to estimate both harms and benefits. One common harm is a false positive (FP), which is a positive screening result, perhaps followed by an invasive test, with no cancer detected on the diagnostic work-up or within a specified time period. An important goal is to estimate the risk of at least one FP, which we call the cumulative risk of an FP, if persons took a regimen of various screening tests, as is commonly recommended. The estimation is complicated because the data come from a study in which subjects are offered various screening tests in rounds with some missing tests in most subjects. Previous methods for estimating cumulative risk of FPs with a single type of test are not directly applicable, so a new approach was developed. METHODS: The tests were ordered by appearance, where the last test was either the first FP (analogous to a failure time) or the last test taken with no FPs having occurred on that test or previously (analogous to a censoring time). We applied a Kaplan Meier approach for survival analysis with the innovation that the hazard for a first FP for a given test depends on the type of test and number of previous tests of that type which were taken. RESULTS: The method is illustrated with data from the screening arm of the randomized Prostate, Lung, Colorectal and Ovarian (PLCO) Cancer Screening Trial. With an FP defined as a diagnostic work-up in the absence of cancer (or advanced adenoma) within three years, the probability of at least one FP among 14 tests in men was 60.5% with 95% confidence interval of (59.3%, 61.6%). CONCLUSION: A simple estimate is proposed for the probability of at least one FP if persons took a regimen of multiple screening tests of different types. The methodology is useful for summarizing the burden of multiphasic screening programmes. PMID- 18416951 TI - Performance of systematic and non-systematic ('opportunistic') screening mammography: a comparative study from Denmark. AB - OBJECTIVES: Evaluation and comparison of the performance of organized and opportunistic screening mammography. METHODS: Women attending screening mammography in Denmark in 2000. The study included 37,072 women attending organized screening. Among these, 320 women were diagnosed with breast cancer during follow-up. Opportunistic screening was attended by 2855 women with 26 women being diagnosed with breast cancer. Data on women attending screening were linked with information on cancer status. Each woman was followed with respect to diagnosis of breast cancer (invasive as well as in situ) for a period of two years. Screening outcome and cancer status during follow-up were combined to assess whether the result of the examination was true-positive, true-negative, false-positive or false-negative. Based on this classification, age-adjusted sensitivity and specificity of organized and opportunistic screening were calculated. RESULTS: Defining BI-RADS(trade mark) 4-5 as a positive screening outcome, the overall sensitivity of opportunistic screening was 33.6% and the specificity was 99.1%. Using BI-RADS(trade mark) 3-5 as positive, the sensitivity was 37.4% and the specificity was 97.9%. Organized screening (which was not categorized according to BI-RADS(trade mark)) had an overall sensitivity of 67.2% and a specificity of 98.4%. CONCLUSION: Our study showed a considerably higher sensitivity in organized screening than in opportunistic screening, while the specificity was fairly similar in the two settings. The findings support implementation of population-based breast screening programmes, as recommended in the 'European guidelines for quality assurance in breast cancer screening and diagnosis'. PMID- 18416952 TI - A cohort effect in cervical screening coverage? AB - OBJECTIVES: The objective of this study is to analyse cervical screening coverage data over time. METHODS: Routinely collected cervical screening statistics, in England, on the proportion of women who have undergone cervical screening with cytology during the preceding five years. The participants included all women residents eligible for cervical screening. RESULTS: Overall coverage remained at about 82% or over between 1995 and 2000. Since 2000, however, coverage has drifted slowly down to just over 80% in 2005. Coverage has long been observed to be related to age. In 2005, the coverage rate was 71% in women aged 25-29, 83% in those aged 35-54 and 75% in those aged 55-64. Comparing coverage by age in the three years--1995, 2000 and 2005--shows broad conformity with this pattern in each of the three years, but overlaid is the fact that at ages below 50, the rate has been falling while at ages above 55, the rate has been rising. The fall in screening coverage appears to be largely a cohort effect, with women born in the 1960s and later being increasingly less likely to participate. CONCLUSIONS: No specific reason for this effect is evident. Action could be targeted at women aged 25-34 to address falling coverage. PMID- 18416953 TI - Incidence of interval breast cancers after 650,000 negative mammographies in 13 Italian health districts. AB - OBJECTIVES: To estimate the total proportional incidence of interval breast cancers in a two-yearly mammography screening programme, and to perform subgroup analyses by woman's age, screening centre-specific recall rate and screening round. METHODS: Using unconverted electronic data-sets from the 13 screening centres in the Emilia-Romagna Region of northern Italy (540,450 women aged 50-69 years), a database of 919,538 mammography records was created. Of these, 655,175 eligible single-mammography records (1997-2002) from 379,318 women were record linked with the regional Breast Cancer Registry. In the two-year inter-screening interval, a total of 1,022,694.3 woman-years at risk were accumulated, with 695 interval cancers observed and 2428.3 expected. The observed number of interval cancers was divided by the expected number to obtain the proportional incidence. RESULTS: The total proportional incidence of first- and second-year interval cancers was 0.18 (95% CI 0.15-0.20) and 0.43 (0.39-0.47), respectively. Woman's age was inversely associated with proportional incidence in both interval years, with a cut-off point at age 60. A screening centre-specific recall rate greater than the regional average of 5% was associated with a proportional incidence of 0.14 (0.11-0.17) versus 0.20 (0.17-0.24) in the first interval year, and of 0.36 (0.31-0.41) versus 0.50 (0.44-0.56) in the second. The proportional incidence remained unchanged between the first and subsequent screening rounds. CONCLUSIONS: The results were in line with the previous Italian data and with the recommended European standards. The inverse effect of woman's age and of recall rate was expected. PMID- 18416954 TI - High-grade cervical abnormalities and screening intervals in New South Wales, Australia. AB - OBJECTIVES: This study aims to determine the association of two versus three year screening intervals with the likelihood of detection of a high-grade cervical abnormality and cervical cancer. METHODS: Data were obtained from the New South Wales (NSW) Papanicolaou (Pap) Test Register (PTR) and NSW Central Cancer Registry (CCR). Subjects were human papillomavirus (HPV) unvaccinated women aged 20-69 years who had a minimum of two Pap tests with a negative result at their first recorded Pap test (n = 1,213,295). Logistic regression was used to determine the association between screening interval and the likelihood of: (1) a cytological prediction of high-grade abnormality, defined as cervical intraepithelial neoplasia (CIN) 2 or greater; (2) a histologically confirmed high grade abnormality; (3) a cytological prediction of cervical cancer and (4) a confirmed diagnosis of cervical cancer, controlling for potential confounders of age and socioeconomic status (SES) of area of residence. RESULTS: For each year increase in the screening interval, the odds of a histologically confirmed high grade abnormality increased significantly in women aged 20-29 years (odds ratio [OR] 1.24, 95% confidence interval [CI] 1.20-1.28) and in women aged 30-49 years (OR 1.11, 95% CI 1.06-1.16), but not in women aged 50-69 years (OR 1.08, 95% CI 0.89-1.32). Similar results were observed for cytologically detected high-grade abnormalities. The screening interval was significantly and positively associated with a cytological prediction of cervical cancer (OR 1.40, 95% CI 1.28-1.54) and a confirmed cervical cancer diagnosis (OR 1.66, 95% CI 1.33-2.07) in women aged 20-69 years. Despite the higher risks, we estimate that if the screening interval were increased from two to three years, and the number of women participating in treatment screening was the same as for biennial screening in NSW, then 140 (95% CI 60-220) fewer high-grade cytology results would occur per year, and there would be around 19 (95% CI -50- 89) fewer high-grade cases confirmed by histology. As the 95% CI covers zero in the latter estimate, there is no significant difference in expected confirmed high-grade abnormalities between biennial and triennial screening if the same number of women are screened triennally as biennially. [corrected] Equivalently, 2.3 (95% CI 1.8-2.8) and 1.9 (95% CI 1.5-2.4) extra cases of high-grade cytology and histology, respectively, would be expected per 1000 women with initially negative cytology if the screening interval were extended from two to three years. CONCLUSION: Increasing the cervical screening interval from two to three years would be expected to significantly increase the odds of detection of a high-grade abnormality for NSW women aged 20-49 years and cervical cancer for NSW women aged 20-69 years. Accordingly, our study provides evidence in support of retaining the recommended cervical screening interval at two years for HPV unvaccinated, well women. PMID- 18416955 TI - Breast cancer screening programme as setting for an adjunct research project: effect on programme attendance. AB - The purpose of this randomized controlled trial was to examine if an adjunct research project to a breast cancer screening programme would affect the high programme attendance. Women residing in the municipality of Tromso aged 55 years or older, scheduled to receive an invitation letter to their first screening round in the Norwegian Breast Cancer Screening Programme during our 15-week recruitment period, were eligible. We randomly selected up to 25 invitees per screening day to receive a mailed request to participate in the research project named the Tromso Mammography and Breast Cancer study. These women constituted the study arm, while the remaining eligible invitees served as the control arm. The attendance rate to the screening programme was 80.1% among the 253 women in the study arm and 74.8% among the 397 women in the control arm (P = 0.09). Our trial finds no effect on the high attendance to the breast cancer screening programme indicating that cancer screening programmes might be suitable settings for adjunct research projects. PMID- 18416956 TI - Routine molecular screening for common alpha-thalassaemia deletions is necessary as part of an antenatal screening programme. PMID- 18416958 TI - Diagnostic accuracy of immunochemical faecal occult blood tests according to number of samples and positivity threshold. PMID- 18416960 TI - Statistical guidelines for authors. PMID- 18416959 TI - Guidance on terminology. PMID- 18416961 TI - Health technology assessment programme. PMID- 18416962 TI - Balloon catheter technology in sinus surgery. PMID- 18416963 TI - Allergic profiles in unilateral nasal polyps of bilateral chronic rhinosinusitis. AB - BACKGROUND: Bilateral chronic rhinosinusitis (CRS), with unilateral nasal polyps (NPs), is a good model for study, being characterized by a similar inflammatory reaction and environment in both nasal cavities of a subject, although NPs exist in only one nasal cavity. It may be feasible to identify specific pathological factors responsible for the disparities by comparing the differences between the nasal cavities. METHODS: In patients with bilateral CRS, polyp tissue and the middle turbinate tissue on the side of the polyp were examined and compared with contralateral nonpolyp middle turbinate tissue. Tissue and serum-specific immunoglobulin E (sIgE) levels were measured, and eosinophilic infiltration, interleukin (IL)-5 and IL-1-beta were assessed also in the polyp tissue. RESULTS: The presence of allergic symptoms correlated well with a positive serum CAP test but poorly with a positive tissue CAP test. There were no obvious differences between tissue sIgE levels on the NP side compared with the nonpolyp side. Tissue eosinophilia was a general characteristic of NPs, whether there was allergy or not. IL-5 and IL-1-beta levels were elevated in NPs, regardless of allergy. There were no differences in the specimens on the nonpolyp side between allergy and nonallergy groups. CONCLUSION: With this model, we have more precise, however, not new results. Localized allergic reaction is not a major factor for the development of a unilateral polyp. Inflammation associated with eosinophilic infiltration and the presence of proinflammatory cytokines, including IL-5 and IL 1-beta, are more important in the pathogenesis of NPs than the allergic reaction. PMID- 18416965 TI - Sinusitis in pilots of different aircraft types: is allergic rhinitis a predisposing factor? AB - BACKGROUND: Despite the high association of allergic rhinitis and acute sinusitis, their exact relationship remains unclear, especially in pilots. The purpose of this study was to analyze the possible relationship of a history of allergic rhinitis with the occurrence of acute sinusitis in Israeli air force pilots. A comparative case series was conducted. METHODS: Events of acute sinusitis were compared between Israeli air force pilots with (n=54) and without (n=82) allergic rhinitis who presented for their annual physical examination. RESULTS: Previous episodes of acute sinusitis were noted in 33% of the pilots with allergic rhinitis and 21% of the control group (p=0.09). A separate analysis of young pilots (<26 years old) yielded corresponding rates of 57% versus 29% (p<0.001). When the groups were divided by type of pilot, the results showed that 54% of the transport pilots, 34% of the fighter pilots, and 13% of the helicopter pilots with rhinitis also suffered from acute sinusitis, as opposed to 28, 15, and 15%, respectively, of the control group. CONCLUSION: Despite careful selection, allergic rhinitis is still a very common disease in pilots and may pose a risk of acute sinusitis. The lower prevalence of acute sinusitis in combat than in transport pilots with rhinitis may be explained by vasoconstriction due to psychological and physiological stress during flight missions. PMID- 18416966 TI - In vitro effects of preservatives in nasal sprays on human nasal epithelial cells. AB - BACKGROUND: The preservatives benzalkonium chloride and potassium sorbate are widely used in nasal drops and sprays. Recently, side effects resulting from mucosal damage caused by benzalkonium chloride and potassium sorbate were reported. METHODS: We investigated the toxicity of benzalkonium chloride and potassium sorbate on human nasal epithelial cells in vitro. Using primary human nasal epithelial cells, different concentrations of benzalkonium chloride, potassium sorbate, or phosphate-buffered saline (PBS; control group) solutions were cocultured with nasal epithelial cells for 15 minutes. Then, the viability of the cells and the cell morphology were assessed. RESULTS: Nasal epithelial cells were more severely damaged with use of clinical preparations or higher concentrations of benzalkonium chloride than in the control group. In addition, nasal epithelial cell membrane lysis was seen on electronic microscopy in the benzalkonium chloride groups. In contrast, there was no significant cell damage seen in the potassium sorbate groups compared with the control group, even with higher concentrations than clinically used. CONCLUSION: Potassium sorbate appears to be a relatively safer preservative than benzalkonium chloride for use in nasal sprays and drops in vitro study. PMID- 18416964 TI - Th2 cytokines associated with chronic rhinosinusitis with polyps down-regulate the antimicrobial immune function of human sinonasal epithelial cells. AB - BACKGROUND: Chronic rhinosinusitis with nasal polyps (CRSwNPs) is a disorder characterized by persistent eosinophilic Th2 inflammation and frequent sinonasal microbial colonization. It has been postulated that an abnormal mucosal immune response underlies disease pathogenesis. The relationship between Th2 inflammatory cytokines and the innate immune function of sinonasal epithelial cells (SNECs) has not been explored. METHODS: Human SNECs (HSNECs) isolated from control subjects and patients with CRS were assessed for expression of antimicrobial innate immune genes and proinflammatory cytokine genes by real-time polymerase chain reaction, ELISA, and flow cytometry. A model of the Th2 inflammatory environment was created by exposure of primary HSNEC to the Th2 cytokine interleukin (IL)-4 or IL-13 for 36 hours, with subsequent assessment of innate immune gene expression. RESULTS: HSNEC obtained from CRSwNP patients displayed decreased expression of multiple antimicrobial innate immune markers, including toll-like receptor 9, human beta-defensin 2, and surfactant protein A. Baseline expression of these genes by normal and CRS HSNEC in culture is significantly down-regulated after incubation with IL-4 or IL-13. CONCLUSION: Expression of multiple innate immune genes by HSNEC is reduced in CRSwNP. One mechanism appears to be a direct effect of the leukocyte-derived Th2 cytokines present in the sinonasal mucosa in CRSwNP. Impaired mucosal innate immunity may contribute to microbial colonization and abnormal immune responses associated with CRSwNP. PMID- 18416967 TI - Influence of essential and fatty oils on ciliary beat frequency of human nasal epithelial cells. AB - BACKGROUND: In alternative and complementary medicine, the use of essential and fatty oils has become more and more popular. In addition to conventional medical therapies, self-medication is showing increasing popularity, using agents with unclear compounds and poorly controlled dosages. Among other disorders, these alternative treatments are used in bronchitis and rhinitis, including some topical applications. Thus, the influence on ciliated epithelia should be evaluated, because a disturbance of the ciliary function can lead to recurrent sinusitis and chronic rhinosinusitis. The aim of this study was to test the influence of fatty and essential oils on the ciliary beat frequency (CBF) of nasal mucosa in vivo. METHODS: The influence of sesame oil, soy oil, peanut oil, Miglyol 840, thyme oil, lavender oil, eucalyptus oil, and menthol on the ciliary activity of nasal brushings was evaluated by digital high-speed imaging. RESULTS: The presence of most fatty oils resulted in an increase in CBF, the effect being highest for peanut oil. Miglyol 840 had no significant influence on CBF. The essential oils were tested at a concentration of 0.2 and 2%. Thyme oil did not affect CBF, whereas the presence of all other essentials oils resulted in an increase in CBF; the effect was higher at 0.2% than at 2%. CONCLUSION: Except thyme oil and Miglyol 840, all tested oils caused an increase in CBF. Interestingly, the 0.2% concentrations of essential oils resulted in stronger effects when compared with the 2% concentrations. PMID- 18416968 TI - Expression of mammaglobins A and B in nasal polyps is similar in patients with and without allergic rhinitis. AB - BACKGROUND: The causes of nasal polyposis remain unclear. Mammaglobins have been implicated in its pathogenesis. However, their association with the occurrence of nasal polyps in the presence of allergic rhinitis (AR) has not been explored. The aim of this study was to compare the expression levels of mammaglobins A and B with the nasal polyps of patients with and without AR. METHODS: Thirty-one patients with bilateral nasal polyposis underwent skin-prick tests to specific aeroallergens. Nasal polyp tissues were obtained from all patients and divided into two groups as nasal polyps with and without AR depending on clinical history and the skin-prick test results. All polyp tissues were analyzed for the levels of mammaglobin A and mammaglobin B by using real-time quantitative polymerase chain reaction technique. RESULTS: Of the 16 samples from patients having nasal polyps with AR, only 1 sample expressed a detectable level of mammaglobin A (1/16). There was no detectable expression of mammaglobin A in tissues from the group of nasal polyps without AR (0/15). Expression of mammaglobin B was detected in all nasal polyp tissues from both groups. The expression of mammaglobin B was not significantly different between nasal polyps with AR (median, 25th-75th percentiles; 0.023, 0.013-0.046) and nasal polyps without AR (0.032, 0.007-0.16). CONCLUSION: Expression levels of mammaglobins A and B in nasal polyps are not different between patients with and without AR. Our findings suggest that mammaglobins' implication in the pathogenesis of nasal polyps is independent of an underlying AR. PMID- 18416969 TI - Serum inflammatory protein profiles in patients with chronic rhinosinusitis undergoing sinus surgery: a preliminary analysis. AB - BACKGROUND: The specific aim of this study was to evaluate surface-enhanced laser desorption/ionization time-of-flight mass spectroscopy (SELDI-TOF-MS) as a clinical screening tool in differentiating patients with chronic rhinosinusitis (CRS) from healthy control patients. METHODS: Serum samples were prospectively collected at a tertiary care academic medical center from 96 CRS patients who underwent functional endoscopic sinus surgery (FESS) and 38 control volunteers who were negatively screened according to the Rhinosinusitis Task Force guidelines on CRS. SELDI-TOF-MS was performed on serum samples to detect protein profiles in the range of 1-100 kDa. Analysis of spectral data was performed to determine the sensitivity and specificity of SELDI-TOF-MS in distinguishing patients with CRS requiring FESS from healthy controls and to determine potential markers for protein purification. RESULTS: Serum protein profiles generated from SELDI-TOF-MS in the range of 1-100 kDa were analyzed. Classification and regression tree analysis based on peak expression correctly classified patients with CRS with 77.1% sensitivity and 65.8% specificity, a positive predictive value of 88%, and a negative predictive value of 53%. Underexpression of a protein peak at 8.4 kDa was associated with CRS in 77% of cases. CONCLUSION: SELDI-TOF-MS serum protein profile analysis is able to distinguish CRS patients requiring FESS from healthy, negatively screened controls with a sensitivity of 77.1% and specificity of 65.8%. Additional investigation is required to determine if SELDI-TOF-MS will make an effective clinical tool. PMID- 18416970 TI - The roles and limitations of computed tomography in the preoperative assessment of sinonasal inverted papillomas. AB - BACKGROUND: Preoperative radiological assessment of sinonasal inverted papilloma (SNIP) is important in the planning of surgical treatment. This study investigates the roles and limitations of preoperative plain computed tomography (CT) scan in the preoperative assessment of SNIP. METHODS: Plain CT scans from 30 patients with SNIP were reviewed retrospectively by a radiologist who had no prior knowledge of the final surgical findings. Disease at each sinus was judged by the CT findings of opacity and additional signs. The overall disease was staged according to the staging system proposed by Krouse. All of the findings were compared with the final disease extent and staging confirmed by intraoperative and histological findings. RESULTS: Using opacity with additional signs for diagnosis, the range of accuracy of CT diagnosis for each sinus involvement was 83-97%. Staging by plain CT was concordant with postoperative staging in 80% of patients. Among the additional signs, focal hyperostosis or "bony strut" had the highest positive predictive value (100%) of tumor origin. CONCLUSION: Focal hyperostosis or bony strut is the most important CT sign predicting the origin of tumor. Although using multiple CT diagnostic signs provides a reasonable assessment of tumor origin and extent, accurate tumor mapping was still impossible because of inadequate differentiation of tumor from inflammatory pathologies. This drawback may be overcome by a complementary MRI scan. Since preoperative CT staging was inaccurate in 20% of cases, surgical planning should be flexible to provide for the need of the intraoperative findings. PMID- 18416971 TI - Utility of preoperative high-resolution CT and intraoperative image guidance in identification of cerebrospinal fluid leaks for endoscopic repair. AB - BACKGROUND: Endoscopic repair of cerebrospinal fluid (CSF) leaks is a well established procedure. The radiological workup and use of intraoperative image guidance (IGS), lumbar drain (LD), and intrathecal fluorescein (IF) are less universally accepted. This study examines endoscopic repair of CSF leaks using preoperative high-resolution CT (HRCT) and intraoperative IGS without IF or LD. METHODS: Retrospective review was performed of a single surgeon experience between September 2002 and January 2007. RESULTS: Forty-two anterior skull base defects in 40 patients were repaired endoscopically. The etiology of CSF leaks was traumatic in 5, spontaneous in 12, and iatrogenic in 25 defects (15 from endoscopic sinus surgeries and 10 from neurosurgical procedures). Fifty-two percent of defects presented with a meningoencephalocele. Defect location was sphenoid sinus (17 defects), ethmoid roof (14 defects), cribriform plate (9 defects), and frontal sinus (2 defects). Defect size ranged from 1x1 mm to 2x2 cm. Follow-up ranged from 8 to 46 months. All defects were identified preoperatively on HRCT with successful intraoperative IGS localization. No patients required IF for identification of CSF leak. Overall successful repair was achieved in 40 (95%) defects after initial repair, and 100% after revision. Seven patients had LD, 6 patients were early in the series. Twenty-three patients were discharged the day after surgery and 6 patients were discharged on day 2. CONCLUSION: Using preoperative HRCT for diagnosis and intraoperative IGS for localization, even small skull base defects can be definitively identified. High success rates and early discharges are achieved without additional procedures such as IF or LD. This is a less invasive and more cost-efficient method of repairing CSF leaks. PMID- 18416972 TI - Orbital and cranial nerve presentations and sequelae are hallmarks of invasive fungal sinusitis caused by Mucor in contrast to Aspergillus. AB - BACKGROUND: Acute fulminant invasive fungal sinusitis (IFS) is a rapidly progressing, destructive process almost exclusively affecting immunocompromised patients. Outcome differences have been found in patients with Mucor and Aspergillus. We performed this study to compare the presenting symptoms and long term morbidity with IFS caused by Mucor versus Aspergillus species. METHODS: Retrospective chart review was performed of 48 patients with 49 cases of acute fulminant IFS over a 19-year period. Presenting symptoms and long-term morbidity related to the orbits and cranial nerves were evaluated. RESULTS: Mucor was found in 22 cases and Aspergillus was found in 27 cases. Orbital (proptosis, periorbital edema, and ophthalmoplegia) and cranial nerve symptoms were seen at presentation more often in Mucor (6 [27%] and 9 [41%]) than in Aspergillus patients (3 [11%] and 7 [26%]; p=0.079). Long-term orbital and cranial nerve sequelae occurred in 16 (72%) Mucor cases and 10 (37%) Aspergillus cases (p=0.0210). The IFS-related mortality was 32% (7) in the Mucor group and 11% (3) in the Aspergillus group (p=0.089). CONCLUSION: Patients with acute fulminant IFS present with similar sinus symptoms; however, there is a trend toward a greater prevalence of orbital and neurological symptoms in patients with Mucor versus Aspergillus. Long-term orbital and neurological morbidity is more prevalent in patients with Mucor compared with Aspergillus. These data suggest that the presence of orbital and neurological symptoms at presentation warrants more aggressive surgical intervention because of the likelihood of Mucor. PMID- 18416973 TI - Effect of postural change on nasal airway and autonomic nervous system established by rhinomanometry and heart rate variability analysis. AB - BACKGROUND: Clinically, nasal obstruction is experienced frequently in the supine position, and the nasal autonomic nervous system (ANS) may be involved in the mechanism. The central ANS functions at maintaining cardiovascular hemodynamics. However, during postural change, the corresponding changes of the central ANS may simultaneously change the nasal airway as well. In this study, the relationships between nasal ANS and central ANS were explored using rhinomanometry (RMM) and heart rate variability (HRV) analysis between postural changes. METHODS: Twelve healthy volunteers aged between 19 and 39 years and without a history of allergic rhinitis or significant nasal anatomic obstruction were enrolled for the study. The nasal airway was measured using RMM in a sitting position and then in a supine position; the electrocardiography was simultaneously recorded. RESULTS: In supine position, the total nasal airflow significantly decreased and the airway resistance significantly increased (p<0.05, Wilcoxon signed-rank test). The ratio of low frequency power to high frequency (HF) power of HRV that represents sympathetic modulation significantly decreased in the supine position (p<0.05, Wilcoxon signed-rank test). However, the HF that represents parasympathetic activity did not show significant change with postural change. The correlations of heartbeat interval with total inspiratory airflow and total inspiratory resistance were significant also (p<0.01, Pearson's correlation). CONCLUSION: The central ANS activities significantly correlated with changes to the nasal airway during postural change. The central ANS, especially the sympathetic nervous system, may play a role in controlling nasal airway during postural change. PMID- 18416974 TI - The use of portable intraoperative computed tomography scanning for real-time image guidance: a pilot cadaver study. AB - BACKGROUND: This study was performed to assess the feasibility of using intraoperative computed tomography (CT) to provide real-time updates to image guidance systems (IGSs) during surgery. METHODS: The xCAT ENT portable intraoperative CT scanner (Xoran Technologies, Ann Arbor, MI) was used to acquire scans before, midway, and at the end of six cadaver dissections during the Southern States Rhinology Course, Augusta, GA, in October 2006. These scans were used to recalibrate three different IGSs used during the dissection. Time measurements were recorded and dosimetry was obtained from the cornea, sphenoid sinus (near the optic chiasm), and from the operative field during acquisition of the images. IGS accuracy was determined at the skull base and lamina papyracea. Surgeons were interviewed on benefits of real-time updates to the IGS after completion of dissections. RESULTS: The xCAT ENT scanner was compatible with all three IGS platforms. The average time to update the IGS was 13 minutes. Radiation doses to the cornea were 620 mrad per scan, and optic chiasm was 800 mrad/scan. The accuracy at the anterior skull base improved from 1.58 to 0.62 mm (p=0.026). The accuracy at the posterior skull base improved from 1.46 to 0.71 mm (p=0.014). The accuracy at the lamina was not significantly changed. CONCLUSION: Intraoperative portable CT scanning with real-time IGS updates is feasible and likely would add little additional time. Accuracy is improved at the skull base. Prospective studies on actual patients are warranted. PMID- 18416975 TI - Use of intraoperative CT scanning in endoscopic sinus surgery: a preliminary report. AB - BACKGROUND: Image-guided functional endoscopic sinus surgery (IG-FESS) using preoperatively acquired CT images for navigation has been proven to facilitate complicated endoscopic sinus procedures and decrease the need for revision procedures. However, this system does not render the intraoperative anatomic changes that occur throughout the procedure. The purpose of this study is to show the technical feasibility and the potential impact of intraoperative CT scanning in conjunction with real-time update of computer-assisted navigation during ESS. METHODS: Intraoperative CT scanning was performed in selected patients undergoing FESS. A scan was taken before the conclusion of surgery performed by skilled endoscopic sinus surgeons. The scans were evaluated for residual disease or cellular partitions. Where indicated, intraoperative scan was then loaded into the image guidance system and intraoperative scans were then used for additional IGS. The scan findings were documented. RESULTS: Intraoperatively acquired CT scans were obtained successfully in <40 seconds and able to be loaded into the image guidance system within minutes. All surgeries were preformed without complication. New information obtained from the intraoperative CT scan led to alteration in the surgical plan in 30% of patients. CONCLUSION: Intraoperative CT scanning can be preformed with currently available technology and has the potential to improve the extent of surgery in patients with complicated anatomy and extensive disease. PMID- 18416976 TI - Long-term endonasal endoscopic review of successful duraplasty after endonasal endoscopic skull base surgery. AB - BACKGROUND: This study was performed to examine the long-term endonasal endoscopic morphological appearance of successful duraplasty after endoscopic skull base surgery for different pathology. METHODS: This study included 65 patients who underwent endonasal endoscopic surgery for different skull base lesions with successful duraplasty. Forty patients had pituitary adenomas, 25 with macroadenomas and 15 with microadenomas. Twenty patients with cerebrospinal fluid rhinorrhea of different etiologies and three patients with meningoencephalocele were included. There were two patients with skull base meningiomas, 1 with an extensive greater wing meningioma reaching the nasal cavity and the 1 with recurrent olfactory groove meningioma. Different types of autologous materials were used in reconstructing the skull base defect. Clinical follow-up with endoscopic nasal examination was done routinely 1, 3, 6, and 12 months after surgery. CT and MRI were performed when indicated. The follow-up period ranged from 6 months to 8 years. RESULTS: Starting from 3 months after surgery to the rest of the follow-up period, endonasal endoscopic view of the site of duraplasty showed that with small skull base defect (<5 mm), there was neither dural pulsation nor prolapse. With moderate-size defect (5-10 mm) there was dural pulsation without prolapse. With larger defect (>10 mm) there was dural pulsation and prolapse. These findings were constant regardless of the etiology of the lesion and the reconstruction material used. CONCLUSION: This long-term study showed that dural pulsation and prolapse at the site of the successful duraplasty is a function of the size of the bony defect and does not depend on the pathology of the lesion or the autologous material used for reconstruction. For any future endonasal procedure for these patients, the surgeons should be fully aware of the state of duraplasty to avoid any complication. PMID- 18416977 TI - The impact of septodermoplasty and potassium-titanyl-phosphate (KTP) laser therapy in the treatment of hereditary hemorrhagic telangiectasia-related epistaxis. AB - BACKGROUND: A variety of modalities are available for the control of recurrent epistaxis in hereditary hemorrhagic telangiectasia (HHT). Laser ablation, in particular potassium-titanyl-phosphate (KTP), has gained popularity as it coagulates the telangiectasia with minimal peripheral tissue injury. Septodermoplasty (SDP) also can be performed in the day surgery setting. Telangiectasia recurred, necessitating repeated treatments. The frequency and interval between procedures is not well documented. The purpose of this study was to describe the frequency of surgical interventions for HHT patients and the impact of SDP. METHODS: A retrospective review was undertaken of procedures performed in a tertiary hospital unit during a 60-month period for HHT. The incidence of KTP laser and SDP, days between treatments, total number of interventions and perioperative hemoglobin (Hb) were audited. Patient groups were identified as definite or possible HHT according to the Curacao criteria. All suffered from epistaxis sufficient to seek medical treatment. RESULTS: Three hundred one procedures were performed on 131 patients during the study period. In total, 33 SDPs and 268 KTP laser treatments were performed; 78.3% of patients required three or less procedures. The mean time interval between treatments was 473 (+/-515 days) days. The rate of KTP after an SDP decreased from 1.83 (+/ 1.99) to 0.78 (+/-0.85; p=0.012). Hb level was not associated with treatments required. CONCLUSION: Surgical interventions with laser and SDP in HHT are always time limited as recurrence of telangiectasia is inevitable. This study discusses the expectations of therapy, mainly frequency and duration of effect, along with the impact of SDP. The outcomes assist in better management of patient expectations. PMID- 18416978 TI - A randomized controlled pilot study of epsilon-aminocaproic acid as a topical hemostatic agent for postoperative bleeding in the sheep model of chronic sinusitis. AB - BACKGROUND: Significant bleeding with blood clot formation in the postoperative period may predispose to the formation of postoperative adhesions. A topical hemostatic agent would potentially improve postoperative comfort and lessen adhesion formation. This pilot study was performed to evaluate the effects of a novel topical hemostatic agent AMICAR (epsilon-aminocaproic acid; Xanodyne Pharmacal, Inc., Florence, KY) on postoperative bleeding after endoscopic sinus surgery (ESS). METHODS: In a prospective randomized controlled pilot study full thickness mucosal injuries were created on the lateral nasal wall, ethmoturbinal and the maxillary ostium on both sides of 10 sheep. Eosinophilic chronic sinusitis was confirmed both by endoscopy and by biopsy before full thickness injuries. The topical hemostatic agent (AMICAR) was sprayed onto a randomly selected side with or without mucoadhesive methyl cellulose (5 sheep in each group). The control side received a spray of saline of equal volume. The degree of bleeding in the surgical field was graded using a visual analog scale until total hemostasis was achieved. The grades at 2-minute intervals as well as the time to achieve total hemostasis were recorded. RESULTS: Eosinophilia of the epithelium did not correlate with the severity of bleeding (p<0.05). There was a statistically significant reduction in the grade of bleeding in the lateral nasal wall both superior and inferior to the middle turbinate attachment when AMICAR alone was used compared with normal saline (p=0.004 and p=0.003, respectively) but when AMICAR was used in combination with methyl cellulose this significance was lost (p=0.076 and p=0.502). CONCLUSION: AMICAR may be considered for use either during or after sinus surgery to reduce bleeding. PMID- 18416979 TI - Effects of sinus surgery on lung transplantation outcomes in cystic fibrosis. AB - BACKGROUND: In cystic fibrosis (CF) patients who are candidates for lung transplant, pretransplant sinus surgery has been advocated to avoid bacterial seeding of the transplanted lungs. This study reviews the 17-year experience of pretransplant sinus surgery among CF patients at a major transplant center. METHODS: Retrospective chart review was performed in all CF patients who underwent heart-lung or lung transplantation at Stanford Medical Center between 1988 and 2005. Postoperative culture data from bronchoalveolar lavage (BAL) and sinus aspirates were evaluated, in addition to survival data. RESULTS: Eighty seven CF transplant recipients underwent pretransplant sinus surgery; 87% (n=59/68) of patients showed recolonization of the lung grafts with Pseudomonas on BAL cultures. The median postoperative time to recolonization was 19 days. Bacterial floras cultured from sinuses were similar in type and prevalence as the floras cultured from BAL. When compared with published series of comparable cohorts in which pretransplant sinus surgery was not performed, there was no statistically significant difference in the prevalence of Pseudomonas recolonization. Times to recolonization also were similar. Survival rates in our cohort were similar to national survival rates for CF lung transplant recipients. CONCLUSION: Despite pretransplant sinus surgery, recolonization of lung grafts occurs commonly and rapidly with a spectrum of flora that mimics the sinus flora. Survival rates of CF patients who undergo prophylactic sinus surgery are similar to those from centers where prophylactic sinus surgery is not performed routinely. Pretransplant sinus surgery does not appear to prevent lung graft recolonization and is not associated with overall survival benefit. PMID- 18416980 TI - Asthma outcomes after endoscopic sinus surgery in aspirin-tolerant versus aspirin induced asthmatic patients. AB - BACKGROUND: Certain diseases affect both upper and lower airways. Aspirin-induced asthma (AIA) is a clinical entity characterized by asthma, nasal polyposis, and aspirin intolerance. To understand the response of the lower airway to surgical treatment of the sinuses, we examined asthma outcomes in AIA compared with a second group of aspirin-tolerant asthmatic (ATA) patients to establish if there were any differences between the two groups after endoscopic sinus surgery (ESS). METHODS: A retrospective record review was performed of 91 asthmatic subjects with chronic rhinosinusitis. Forty-one subjects had AIA and 50 subjects had ATA. Subjective and objective asthma outcome parameters were used to compare between the two groups at three time points: immediately before ESS and 6 and 12 months after ESS. RESULTS: Preoperatively, AIA patients had significantly higher asthma severity (p<0.0001) and lower forced expiratory volume in 1 second values (p=0.04). At 12 months after ESS, a statistically significant difference between the two groups with better results in AIA patients was seen in asthma severity improvement (p=0.010) and in the decrease of ICS doses (p<0.0001), without significant differences between the two groups in other asthma outcome parameters. CONCLUSION: AIA patients usually present with more severe asthma. The asthmatic complaints of AIA and ATA patients continue to improve significantly over 6 and 12 month after ESS. Although ESS helped both groups of patients, AIA had statistically significant better results compared with ATA patients in asthma severity scores and decreased need for ICS. PMID- 18416981 TI - Functional endoscopic dilatation of the sinuses: patient satisfaction, postoperative pain, and cost. AB - BACKGROUND: The purpose of this study was to determine how functional endoscopic dilatation of the sinuses (FEDS) compares with functional endoscopic sinus surgery (FESS) in a select group of patients with respect to (1) elimination of symptoms, (2) patient satisfaction, (3) postoperative narcotic use, and (4) cost. A retrospective study was performed of 70 patients with chronic rhinosinusitis who underwent FEDS or FESS as primary or revision treatment. METHODS: Symptoms and satisfaction based on the Sino-Nasal Outcome Test (SNOT-20) questionnaires and global patient assessment, postoperative narcotic use, and costs were compared after 3-month follow-up. RESULTS: SNOT-20 change scores indicated that both FEDS and FESS had clinically meaningful treatment responses. Patient satisfaction was higher and postoperative narcotics usage was less with FEDS. The cost for primary procedures was similar, whereas the cost for revision surgery using FEDS was considerably less. Turbinate lateralization and scarring was more common in the FEDS group, particularly early in the study. The incidence of recurrent sinus infections during the follow-up period was similar for both groups. Only one patient in the FEDS group required a repeat intervention within the short-term follow-up period. CONCLUSION: Both FEDS and FESS resulted in significant improvement in SNOT-20 scores for selected patients with mild disease. Patient satisfaction and postoperative narcotic use of FEDS compare favorably with FESS. Cost of FEDS was comparable with FESS for primary procedures but was less than FESS for revision procedures. Long-term efficacy and final cost of FEDS remain to be addressed, taking into account the need for revision procedures after initial FEDS, by means of long-term studies and objective outcome measures. PMID- 18416982 TI - Outcomes of posterior lacrimal sac approach in endoscopic dacryocystorhinostomy: review of 35 cases. AB - BACKGROUND: The purpose of this study was to describe the posterior lacrimal sac approach in endoscopic dacryocystorhinostomy (DCR) performed at our institute and report perioperative results achieved with this procedure. METHODS: A prospective clinical study was performed of 35 adult patients with nasolacrimal duct obstruction who underwent posterior lacrimal sac approach DCR from March 1998 to May 2005. Follow-up period ranged from 13 to 30 months (average, 17.5 months; SD, 6.8 months). "Surgical success" was defined as complete relief of epiphora and patent surgical ostium on endoscopic assessment. RESULTS: Surgical success was achieved in 30/35 (85.7%) patients after the primary surgery. Of 5 unsuccessful patients who complained of occasional epiphora, 4 patients had formed stenosis and 1 patient had granulation around the surgical opening, and all had revision surgery. Four of 5 (80.0%) patients achieved surgical success. Thus, including the result of revision surgery, 34/35 (97.1%) patients were successful. CONCLUSION: Posterior lacrimal sac approach in endoscopic DCR has several advantages: good sac accessibility, a low complication rate, and a relatively high success rate. Therefore, the posterior lacrimal sac approach appears to offer a useful alternative approach for the surgical treatment of nasolacrimal duct obstruction. PMID- 18416983 TI - Silicone tubing is not necessary after primary endoscopic dacryocystorhinostomy: a prospective randomized study. AB - BACKGROUND: Endoscopic dacryocystorhinostomy (EN-DCR) is an effective procedure when treating saccal and postsaccal obstructions of the nasolacrimal pathway. The benefit of silicone tubing after DCR is still controversial. We conducted a prospective, randomized study to evaluate the necessity of bicanalicular silicone tubes after primary EN-DCR. METHODS: Forty-six consecutive primary EN-DCR procedures were performed in 42 patients during 2004-2007. The patients were randomized into two study groups according to whether silicone tubing was used or not. RESULTS: The overall success rate after primary EN-DCR was 89%: with silicone tubes it was 78%, and without silicone tubes it was 100%. The difference between these two groups was statistically significant (p<0.049). The follow-up period was 6 months and included three follow-up visits: 1 week, 2 months, and 6 months postoperatively. If inserted, the silicone tubes were removed at the 2 month visit. CONCLUSION: The results of our prospective, randomized study showed that the use of silicone tubes after primary EN-DCR is not necessary. PMID- 18416985 TI - Promotion of oral health: need for interprofessional collaboration. PMID- 18416986 TI - Upfront. Avocados may help prevent oral cancer, OSU study shows. PMID- 18416988 TI - Effects of a stannous fluoride-impregnated dental floss on in vivo salivary fluoride levels. AB - PURPOSE: The aim of this in vivo pilot study was to determine the concentration of fluoride retained intra-orally in saliva after flossing with dental floss impregnated with stannous fluoride (SnF(2)). METHODS: Participants flossed their teeth ad libitum with 2 premeasured lengths of fluoridated dental floss. Expectorated saliva samples were collected in vials before flossing (PF), immediately postflossing(IPF), at 30 minutes (30), and 1 hour (60) after flossing for analysis with a fluoride-specific electrode and an Orion millivoltmeter. Postflossing samples were compared to the preflossing samples using ANOVA and Tukey's HSD. RESULTS: Differences between the PF and IPF group means were found to be statistically significant at p<0.01. No other significant differences were found between or among any of the groups. Salivary fluoride levels at 60 minutes (60) were similar to those prior to flossing (PF). CONCLUSIONS: It can be concluded that fluoride can be released from flossing with the tested SnF(2) impregnated dental floss elevating salivary fluoride levels for at least 30 minutes. Use of this fluoride-containing dental floss offers an option for delivery of fluoride to individuals at risk for dental caries. PMID- 18416990 TI - Moving research knowledge into dental hygiene practice. AB - Dental hygiene, as an emerging profession, needs to increase the number of intervention studies that identify improvements in oral health outcomes for clients. Historically, dental hygiene studies have typically been atheoretical, but the use of theoretical frameworks to guide these studies will increase their meaningfulness. Rogers' theory of diffusion of innovations has been used to study research utilization across many disciplines, and may offer insights to the study of research use in dental hygiene. Research use is an important component of evidence-based practice (EBP), and diffusion of research knowledge is an important process in implementing EBP. The purpose of this paper is to use diffusion of innovations theory to examine knowledge movement in dental hygiene, specifically through the example of the preventive practice of oral cancer screening by dental hygienists, considered as an innovation. Diffusion is considered to be the process by which an innovation moves through communication channels over time among a social network. We suggest diffusion theory holds promise for the study of knowledge movement in dental hygiene, but there are limitations including access to and understanding research studies as innovations. Nevertheless, using a theoretical framework such as Rogers' diffusion of innovations will strengthen the quality of intervention research in dental hygiene, and subsequently, health outcomes for clients. PMID- 18416989 TI - Survey of oral health practices among adults in a North Carolina Hispanic population. AB - PURPOSE: The purpose of this study was to collect baseline data pertaining to the oral health of Hispanics residing in Siler City, NC, a microcosm of the flourishing Latino growth found especially in the southeastern United States. METHODS: A convenience sample of 158 Hispanic adults was recruited. A 41-item Spanish language survey was utilized. Questions addressed preventive oral health practices, oral health knowledge and beliefs, perceived needs, and demographic information. RESULTS: Analysis of data were conducted to find demographic characteristics, frequencies, and correlations. The following associations were found statistically significant (p < .05): (1) brushing frequency and belief that healthy gums bleed; (2) dental insurance and frequency of dental visits; (3) source of oral hygiene instruction and participant's reported brushing or flossing, and dental visit frequency. CONCLUSIONS: This exploratory study found that this population lacks knowledge related to oral health and that they have indicated an overwhelming need for preventive dental prophylaxis. Characteristics found in this population were low income, low education, no dental insurance, and not speaking fluent English. The following suggestions may aid in eliminating dental health care disparities among this population: 1) training and recruiting oral health care providers that are culturally sensitive and speak Spanish; 2) conducting oral health promotional programs sensitive to the needs of Hispanics; 3) Dental Hygiene programs should expand clinical rotations to provide culturally diverse dental hygiene services; 4) changes should be made in North Carolina dental hygiene practice laws to increase care to underserved population. PMID- 18416991 TI - Advanced caries in a patient with a history of bariatric surgery. AB - PURPOSE: The rate of bariatric surgery in the United States has risen significantly in the past decade as the prevalence of extreme obesity continues to increase. Although bariatric surgery is an effective therapeutic modality for extreme obesity, it is associated with risk factors and select oral health conditions. This case study describes a patient with a significant increase in dental caries after having gastric bypass surgery. It was hypothesized that bariatric patients may be at an increased risk for caries due to the need for more frequent and prolonged meals throughout the day. METHODS: Dietary, bacterial, and salivary risk assessments were completed 5 years postsurgery. A 7 day food record was analyzed using Nutrition Calc Plus + 2.0 software and a plaque index was measured. In vitro test kits were used to evaluate the cariogenicity of the plaque as well as the quantity, quality, and buffering capacity of the patient's saliva. Clinical exams and dental radiographs were used to evaluate caries activity pre-and post-surgery. RESULTS: On average, the patient typically consumed fermentable carbohydrates 5 times per day with 12% of her calories from sucrose. The plaque index and cariogenicity were high. The patient demonstrated low salivary production with poor buffering capacity and viscous saliva. The caries activity was noticeably higher postsurgery versus presurgery. CONCLUSION: The present case study supports the hypothesis that recommended postsurgical meal patterns may place the patient at an increased risk for dental caries, particularly in the presence of other risk factors. More research is needed to further evaluate this relationship. PMID- 18416993 TI - Is carotid artery stenting safer and more effective than endarterectomy? PMID- 18416992 TI - A much-anticipated FDA reform bill is signed into law: progress and compromise. PMID- 18416994 TI - Carotid endarterectomy or carotid artery stenting? PMID- 18416995 TI - Update on antiplatelet agents, including MATCH, CHARISMA, and ESPRIT. AB - Despite recent advances in the acute treatment of stroke, prevention and risk factor modification remain the mainstays of management for patients with ischemic stroke and transient ischemic attack. The majority of noncardioembolic ischemic strokes are atherothrombotic, presumed to be associated with the activation and aggregation of platelets. Antiplatelet medications have been shown to be effective in the secondary prevention of stroke of presumed arterial origin, both as monotherapy and in combination. Among combination of antiplatelet agents, aspirin plus extended-release dipyridamole has demonstrated statistically significant additive benefit over monotherapy with each agent. Clopidogrel plus aspirin does not prevent recurrent ischemic stroke over each component individually, and the combination increases the risk of hemorrhagic side effects. This article reviews the most recent studies on antiplatelet medications, including the combination of aspirin and clopidogrel or extended-release dipyridamole, and discusses some of the controversies that still exist with the use of antiplatelet agents. PMID- 18416996 TI - Indications for carotid artery surgery and stent: the role of carotid ultrasound. AB - Carotid artery disease is the most frequently identified cause of ischemic stroke and is mostly due to atherosclerotic disease. Landmark trials have demonstrated that surgical intervention in cases of high-grade carotid stenosis can reduce the risk of subsequent stroke. Endovascular approaches continue to be evaluated in ongoing trials. Careful patient selection is critical if the potential benefits of carotid revascularization are to be realized. Ultrasound is a safe, accurate, readily available method to evaluate carotid artery disease. The degree of stenosis is the parameter most frequently used to make decisions about therapeutic approaches. Plaque characteristics may also be useful for identifying high-risk patients. Microembolic signals detected by transcranial Doppler ultrasound can identify cerebral embolization before or after carotid intervention. This review discusses the current clinical role of carotid ultrasound in the selection of patients for the two most frequently used carotid interventions: carotid endarterectomy or carotid angioplasty and stenting. PMID- 18416997 TI - Carotid plaque, stroke pathogenesis, and CRP: treatment of ischemic stroke. AB - C-reactive protein (CRP), an inflammatory marker, has been identified as a likely predictor of the risk of a future stroke. In clinical settings, it has been consistently observed that higher concentrations of CRP are associated with larger brain infarcts and worst neurologic outcome. However, there is still controversy over the degree of risk conferred by elevated CRP concentrations. CRP, like many other hemostatic factors, is an acute-phase protein and, therefore, it is not always clear whether its association with cerebrovascular disease reflects its contribution to atherothrombosis, its acute-phase condition, or both. Whether a reduction of CRP levels could be beneficial to stroke patients remains to be clarified. More studies are needed before CRP becomes a routine part of the evaluation of stroke patients. PMID- 18416998 TI - Aortic occlusion for cerebral ischemia: from theory to practice. AB - Ischemic stroke is a devastating disorder with limited treatment options. Arterial recanalization with thrombolysis or mechanical thrombectomy may be used to restore perfusion in only a subset of cases. Collateral therapeutic strategies such as partial aortic occlusion attempt to reverse ischemia, the principal detrimental element in stroke pathophysiology. This article considers the theoretic basis of aortic occlusion as a therapeutic strategy for cerebral ischemia, procedural details employing the NeuroFlo (CoAxia, Maple Grove, MN) device, ongoing and prior clinical studies, and potential practice implications in the future. The hemodynamic mechanisms associated with flow redistribution due to aortic occlusion and impact on the dynamic role of collateral perfusion in the ischemic brain are considered. Targeting ischemia rather than clot disruption or consideration of venous hemodynamics and flow redistribution may initiate a radical transformation in stroke care. Ultimately, demonstration of a rational mechanism that averts ischemia will be essential. PMID- 18416999 TI - Neuroprotection in cerebral ischemia: emphasis on the SAINT trial. AB - Acute ischemic stroke (AIS) is a significant cause of death and disability in the United States. It has been 10 years since tissue plasminogen activator became the first medication approved by the US Food and Drug Administration for treatment for AIS. However, this treatment simply reopens arteries. The identification of deleterious cellular reactions that occur secondary to cerebral ischemia has led investigators to search for neuroprotection strategies to complement reperfusion. More than 100 human trials, including a handful of phase III trials, had failed to produce an efficacious neuroprotective agent. In 2006, the first positive trial of neuroprotection was published: the SAINT I (Stroke-Acute Ischemic NXY Treatment) study. In February 2008, the SAINT II study was published, indicating that NXY-059 was not effective for AIS treatment. PMID- 18417000 TI - Positron emission tomography for perfusion and function. PMID- 18417001 TI - The promise of whole-heart coronary MRI. AB - The noninvasive detection of coronary artery disease has been a major goal of newer cardiac imaging technologies. Over the past decade, coronary MRI has undergone significant advances, resulting in excellent sensitivity for detecting coronary artery disease. Whole-heart coronary MRI, a technique that is somewhat analogous to coronary CT angiography, has emerged as a promising approach for the noninvasive evaluation of the coronary arteries. PMID- 18417002 TI - Developments in coronary CT angiography. AB - CT imaging of the coronary arteries by contrast-enhanced CT, termed "coronary CT angiography" or "coronary CTA," has become increasingly stable and robust during the past few years. Several trials have demonstrated rather high diagnostic accuracies in somewhat selected patient groups, and this method can be applied clinically to rule out coronary artery stenoses. Based on clinical considerations, this will most likely be beneficial for symptomatic patients who are not at high risk for coronary artery disease, both with stable symptoms or presenting with acute chest pain. Other applications are more problematic, such as use in patients after stent placement or coronary artery bypass graft surgery. Further improvements in technology are expected to allow expansion of indications for coronary CTA in the future. PMID- 18417003 TI - Epidemiologic guidance with coronary artery calcium scoring. AB - The importance of screening for subclinical coronary artery disease is reinforced by the detection gap existing between the currently used risk stratification tools and the persistently elevated rates of cardiovascular disease in Western countries. Medicare data clearly indicate the extremely high cost of caring for patients with end-stage diseases, and early detection may curb some of these expenses. Coronary artery calcium screening has become a widely used tool to estimate risk in a variety of categories in the general population and is discussed in this review. PMID- 18417004 TI - Assessment of ventricular structure and function with multidetector CT and MRI. AB - Accurate, noninvasive assessment of ventricular function is fundamental to providing excellent care to patients with cardiovascular diseases. Three dimensional imaging using cardiac magnetic resonance (CMR) permits the measurement of ventricular structure and function with such precision and accuracy that it now serves as the standard of reference for this purpose. Multidetector CT (MDCT) permits similar three-dimensional reconstruction and measurement of ventricular function. Available data indicate there is good agreement between MDCT and CMR measurements of ventricular function. Patients with cardiomyopathy and distorted ventricles stand to benefit the most from these techniques, particularly with the possibility of combined noninvasive angiography and systolic function assessment with MDCT. PMID- 18417005 TI - Late gadolinium-enhanced cardiac magnetic resonance. AB - Recent advances in cardiovascular magnetic resonance have made it an important clinical tool. Late gadolinium enhancement imaging after gadolinium administration is now a well-validated and practical test to assess myocardial scarring. Due to its high spatial resolution, late gadolinium enhancement is much more sensitive in detecting small infarctions than older imaging modalities. However, late gadolinium enhancement is not specific for myocardial infarction but can occur in a number of cardiomyopathies. New studies suggest that the extent and pattern of enhancement may have prognostic and therapeutic implications, making it an important non-invasive instrument in cardiovascular diagnostics. PMID- 18417006 TI - Courage under fire: what is the optimal approach to initial treatment of stable angina? PMID- 18417007 TI - What is new in the 2006 ACC/AHA guidelines on valvular heart disease? AB - In June 2006, the American College of Cardiology/American Heart Association Guidelines for the Management of Patients with Valvular Heart disease were published. Although many recommendations were unchanged from the 1998 guidelines, several new suggestions bear comment. The new guidelines favor a more aggressive approach to most valve lesions. For patients with low-gradient, low ejection fraction aortic stenosis, detecting inotropic reserve to help risk stratify such patients is now recommended. In the field of aortic regurgitation, the use of vasodilators to forestall surgery has been called into doubt by newer data; thus, recommendations for vasodilator use have been softened. For patients with mitral regurgitation, earlier surgery and mitral repair are highlighted. The new guidelines also reflect a growing trend toward the use of biological valves instead of mechanical prostheses. This 150-page document with more than 1000 references should be useful in guiding practice for these often complex patients. This article summarizes some of the more salient changes in 2006 compared with those of 1998. PMID- 18417008 TI - New approaches to quantifying aortic stenosis severity. AB - Previously, aortic valve stenosis (AS) etiology was usually congenital or due to rheumatic disease. However, the most frequent cause is now degenerative AS, which is often part of a continuum including increased rigidity of the aorta due to atherosclerosis and left ventricular dysfunction due to coronary artery disease. This article highlights newer approaches to quantify AS taking into account the inter-relation between the different components (valvular, vascular, and ventricular) affecting clinical outcome in these patients. Emphasis is given to a more comprehensive evaluation of AS severity going beyond classical measurements and including indices such as 1) the energy loss index to quantify the valvular obstruction net of pressure recovery; 2) systemic arterial compliance to quantify vascular load; and 3) valvular-arterial impedance to assess the global (valvular + vascular) increase in afterload. Routine use of these indices, easily measured by Doppler echocardiography, should improve clinical management of AS patients. PMID- 18417009 TI - Imaging modalities in valvular heart disease. AB - Evaluating valvular heart disease requires a multi-parametric analysis of valvular pathology, hemodynamic derangements, and impact on ventricular size and function. The capability to perform real-time three-dimensional (3-D) imaging has vastly strengthened the already established role of echocardiography. CT and MRI advances have led to their use as daily clinical tools. Two-dimensional and 3-D echocardiography and Doppler modalities allow for accurate assessment of valvular lesions, pressure gradients, stenotic valve orifice areas, pulmonary artery pressures, intracardiac pressures, and regurgitant volumes. Quantitation of chamber volumes has become more accurate and reproducible with 3-D echocardiography, CT, and cardiac MRI. Although ultrasound imaging is the primary tool, the other techniques provide adjuvant or alternate options to examine valvular heart disease. This array of imaging modalities is likely to provide greater insights into the pathophysiology of valvular heart disease, new pointers to prognosis, and also guide innovative treatment strategies. PMID- 18417010 TI - Percutaneous aortic valve replacement. AB - Aortic valve replacement can produce dramatic benefit in the setting of aortic stenosis. The potential for morbidity and mortality associated with thoracotomy, cardiopulmonary bypass, and aortotomy has fostered a search for alternatives. Early experience with transcatheter endovascular aortic valve implantation demonstrated feasibility and durability, but the procedure was difficult to reproduce. However, equipment, techniques, and experience have evolved rapidly. Balloon-expandable and self-expanding prostheses and percutaneous femoral artery and open left ventricular apical access have found favor, each with potential advantages and disadvantages. Procedural success rates and clinical outcomes continue to improve. Current studies suggest that morbidity and mortality are competitive in comparison to conventional surgery in selected high-risk patients. PMID- 18417011 TI - Tricuspid valve repair for treatment and prevention of secondary tricuspid regurgitation in patients undergoing mitral valve surgery. AB - Secondary or functional tricuspid regurgitation occurring late after mitral valve surgery is associated with high morbidity and mortality. In this article, we review the pathophysiology of secondary tricuspid regurgitation and the evidence supporting the use of tricuspid valve annuloplasty for preventing and treating secondary tricuspid regurgitation. Liberal application of tricuspid valve annuloplasty is recommended to prevent progression of secondary regurgitation, as contrary to widely held opinion, fixing the left-sided valve dysfunction often does not resolve secondary tricuspid valve dysfunction. Based on existing literature, assessing the tricuspid valve annular dimensions can be recommended as part of all mitral valve operations, and annuloplasty strongly considered in patients with tricuspid annular dilatation or moderate to severe tricuspid regurgitation. PMID- 18417012 TI - Molecular imaging of atherosclerosis. AB - Techniques are being developed for clinical molecular imaging of atherosclerosis to identify and characterize vulnerable plaques in each vascular territory. Molecular imaging encompasses multiple imaging modalities that depict cellular and subcellular processes. Molecular imaging can provide a "virtual histology" noninvasively about atherosclerotic disease, characterizing vascular lesions with markers of inflammation, angiogenesis, lipid metabolism, and more. PMID- 18417013 TI - Targeting tissue angiotensin-converting enzyme for imaging cardiopulmonary fibrosis. AB - Convincing clinical and experimental evidence exists that the renin-angiotensin system and its primary effector peptide, angiotensin II, are linked to the pathophysiology of interstitial fibrosis in the heart and in the lung. Tissue angiotensin-converting enzyme and locally synthesized angiotensin II exert local trophic effects, which regulate fibroblast growth and proliferation. Thus, the presence of tissue renin-angiotensin system offers an opportunity for targeted imaging, which may be valuable for understanding the pathobiology of cardiopulmonary disorders and for guiding medical therapy. PMID- 18417014 TI - Advances in rubidium PET and integrated imaging with CT angiography. AB - The role of rubidium-82 (Rb-82) positron emission tomography (PET) in the evaluation and care of patients with suspected coronary artery disease is evolving in conjunction with advances in PET instrumentation, data analysis, and clinical research. Instrumentation developments such as three-dimensional acquisition, new scintillator materials, and x-ray CT help to improve the quality of Rb-82 images. New approaches to kinetic modeling and software tools for analysis of clinical Rb-82 studies are being developed and evaluated, enabling quantification of absolute myocardial blood flow and flow reserve. Recent clinical research studies are providing new insights into the value of Rb-82 cardiac imaging compared with single photon emission CT myocardial perfusion imaging. Integrated x-ray CT angiography and Rb-82 PET perfusion imaging on hybrid PET/CT systems is an exciting new prospect. Complementary anatomical and functional information on atherosclerosis and ischemia can be provided in a single imaging session for better diagnosis and risk stratification of patients with coronary artery disease. PMID- 18417015 TI - Myocardial fatty acid metabolism and cardiac performance in heart failure. AB - It is well established that cardiac metabolism is abnormal in heart failure (HF). Experimental studies suggest that in severe HF, cardiac metabolism reverts to a more fetal-like substrate use characterized by enhanced glucose and downregulated free fatty acid (FFA) metabolism. Correspondingly, in humans, when FFA levels are similar, myocardial glucose metabolism is increased, and FFA metabolism is decreased. However, depression of left ventricular function and insulin resistance induces a shift back to greater FFA uptake and oxidation by increasing circulating FFA availability. Myocardial insulin resistance may further impair myocardial glucose uptake and lead to an energy depletion state. Experimental and preliminary clinical studies suggest that metabolic modulators enhancing myocardial glucose oxidation may improve cardiac function in patients with chronic HF. However, it has been found that acute FFA deprivation is harmful to the cardiac performance. Optimizing myocardial energy metabolism may serve as an additional approach for managing HF, but further studies are warranted. PMID- 18417016 TI - The role of integrated PET-CT scar maps for guiding ventricular tachycardia ablations. AB - Reentrant ventricular tachycardia is the next emerging frontier in electrophysiology. Current ablation strategies rely on endocardial voltage measurements to identify myocardial scar and guide catheter ablation procedures. However, this voltage mapping approach has several inherent limitations. In patients with structural heart disease, positron emission tomography (PET)/CT has the potential to provide supplementary scar characterization by displaying additional metabolic (by PET) and morphologic (by CT) tissue-specific information. Three-dimensional scar maps can be created from the imaging datasets, which are uploaded into clinical mapping systems, and can facilitate substrate-guided ablation procedures. This has the potential to shorten procedure times, decrease complications, and improve the procedural success. PMID- 18417017 TI - Neurolytic blocks revisited. AB - The use of neurolytic blockade is a staple in the management of cancer pain. However, the data on neurolysis for chronic pain are plagued by inconsistencies in patient selection, diagnostic criteria, technical standards, and outcome measures. No one neurolytic agent or technique has been proven superior to another. Current evidence suggests that patients with pain of malignant origin may benefit from a variety of neurolytic techniques, as the benefit of documented short-term pain relief may outweigh risk at the end of life. In the absence of compelling data suggesting low-risk long-term efficacy, neurolysis for chronic benign pain should be cautiously considered, in most cases, only after failure of aggressive multidisciplinary management. PMID- 18417018 TI - Intradiscal electrothermal therapy, percutaneous discectomy, and nucleoplasty: what is the current evidence? AB - Over the past decade, there has been a surge of minimally invasive techniques aimed at treating both discogenic low back pain (LBP) and radicular pain. This article assesses the current evidence for three such treatments: intradiscal electrothermal therapy (IDET), percutaneous discectomy, and nucleoplasty. An electronic search of the literature carried out using the Cochrane Library database (2007) and Medline (1966-2007) identified 77 references relating to IDET, 363 to percutaneous discectomy, and 36 to nucleoplasty. Two randomized controlled trials (RCTs) assessed the effectiveness of IDET; one demonstrated a positive effect on pain severity only, whereas the other demonstrated no substantial benefit. Other RCTs show that percutaneous intradiscal radiofrequency thermocoagulation is ineffective for the treatment of discogenic LBP. Trials of automated percutaneous discectomy suggest that clinical outcomes after treatment are at best fair and often worse when compared with microdiscectomy. There are no published RCTs assessing Coblation (ArthroCare Spine, Stockholm, Sweden) technology. PMID- 18417019 TI - Kyphoplasty and vertebroplasty. AB - Percutaneous vertebroplasty is the injection of a vertebral compression fracture (VCF) with bone cement, generally polymethylmethacrylate. Percutaneous kyphoplasty is the placement of balloons into the vertebral body with an inflation/deflation sequence to create a cavity before the cement injection. These procedures are most often performed in a percutaneous fashion on an outpatient (or short stay) basis. The procedure is indicated for painful VCFs due to osteoporosis or malignancy, and painful hemangiomas. The procedure may have efficacy in painful vertebral metastasis and traumatic compression fractures. Much evidence favors the use of this procedure for pain associated with these disorders. The overall risks of the procedure are low, but serious complications (including spinal cord compression) can occur. With good patient selection and careful technique, these complications are avoidable, making the risk-to-benefit ratio highly favorable. PMID- 18417020 TI - Peripheral nerve stimulation for chronic pain. AB - Electrical stimulation has been used since ancient times to treat painful conditions. Electrotherapy for pain was largely consigned to the realm of quackery until the introduction of the Gate Control Theory by Melzack and Wall in 1965 provided a rationale for direct stimulation of peripheral nerves. Since that time, peripheral nerve stimulation has been applied to the treatment of painful conditions throughout the body, beginning with the major nerves of the extremities and culminating today in precise subcutaneous field stimulation targeted to specific areas of neuropathic pain. This article reviews the history, development, and current areas of interest in peripheral nerve stimulation for the treatment of neuropathic pain. PMID- 18417021 TI - Cognitive effects of opioids. AB - Although opioid analgesics are safe and effective tools for the treatment of moderate to severe pain, there remain large gaps in understanding of the effects of these drugs on the many dimensions of functioning. This article summarizes the biomedical evidence addressing cognitive effects of the opioid analgesics. Current evidence indicates that cognitive function can be influenced by use of opioid analgesics, although the effects vary between drugs, are thought to be most significant with mixed-activity drugs, codeine, propoxyphene, and meperidine, and are generally most concerning during the first few days after starting opioid therapy, before tolerance develops. Blanket policies regarding the activities of driving and working are inappropriate; this issue is best addressed on a patient-specific basis. PMID- 18417022 TI - Pulsed radiofrequency for chronic pain. AB - Pulsed radiofrequency (PRF), a technology related to continuous radiofrequency, is unique in that it provides pain relief without causing significant damage to nervous tissue. The mechanism by which PRF controls pain is unclear, but it may involve a temperature-independent pathway mediated by a rapidly changing electrical field. Although much anecdotal evidence exists in favor of PRF, there are few quality studies substantiating its utility. PMID- 18417023 TI - Chronic daily headache: when to suspect sinus disease. AB - Migraine may be responsible for many headaches thought to be caused by the sinuses. Patients complaining of "sinus headache" must have a complete ear, nose, and throat examination. Occasionally, chronic headaches may arise from the sinuses; a thorough history is important to search for symptoms of facial pain and pressure along with other nasal sinus symptoms. PMID- 18417024 TI - Chronic daily headache: when to suspect meningitis. AB - Millions of patients see physicians each year for headache, most of which are primary headaches. However, serious secondary headaches, such as meningitis, represent about 5% of children and 1% to 2% of adults seen in the emergency department for headache. A primary care or emergency department physician may initially miss individuals with bacterial meningitis. Considering meningitis as a headache cause is important because delay in the diagnosis may have adverse consequences. A careful history and physical examination are central in identifying individuals at high risk for meningitis. This article lists information that can be obtained from the patient that may be indicative of meningitis. Performing a lumbar puncture with appropriate examination of the cerebrospinal fluid (CSF) is the key to establishing the diagnosis of meningitis. This article also includes the types of meningitis that should be considered when the CSF demonstrates a pleocytosis. PMID- 18417025 TI - Obesity and chronic daily headache. AB - Population studies suggest that obesity is associated with migraine progression from episodic to chronic daily headaches. Although not a risk factor for episodic migraine, obesity is associated with frequent and severe headaches among migraineurs. We suggest that obesity is an exacerbating factor for migraine. This association seems to be specific to migraine and does not apply to tension-type headache. We review the clinical evidence that links obesity and migraine progression. We then review the potential pathophysiologic mechanisms to support this relationship. We close by briefly discussing clinical interventions related to obesity and migraine. PMID- 18417026 TI - Occipital nerve stimulation for medically intractable headache. AB - There is an unmet need, and thus a continued search, for effective treatments for patients with chronic daily headache who do not respond to or tolerate conventional therapies. Recent interest has focused on the use of occipital nerve stimulation (ONS) to treat patients with chronic, disabling headaches who are not adequately controlled by usual therapies. A small but growing body of evidence supports the use of ONS for the treatment of intractable headache. Electrical stimulation of the occipital nerve has both peripheral and central nervous system effects that modulate nociception. The exact role for ONS will evolve as our understandings of its physiologic effects, efficacy, and safety grow. PMID- 18417027 TI - Chronic post-traumatic headache: associations with mild traumatic brain injury, concussion, and post-concussive disorder. AB - Chronic post-traumatic headache (PTH) always arises from some sort of head injury such as mild traumatic brain injury or concussion. The terminology and definitions for these injuries remain controversial and continue to evolve. This article reviews and updates current terminology and definitions for these head injuries, along with current concepts of pathophysiology. The second portion of this article considers the persistence of the other symptoms associated with chronic PTH, which constitute the post-concussive disorder. PTH is the most common symptom after a head injury, but post-concussive symptoms often complicate the clinical picture and must be considered in the overall management of the chronic PTH patient. PMID- 18417028 TI - Chronic headache and pituitary tumors. AB - Pituitary tumors come to clinical attention due to endocrine dysfunction, distortion of local structures surrounding the pituitary fossa, or as an incidental finding during neuroimaging for headache. Explanations for pituitary tumor-associated headache include stretching of the dura mater and invasion of pain-producing structures within the cavernous sinus. However, small functional pituitary lesions may present with severe headache without cavernous sinus invasion or suprasellar extension. Prolactinomas and growth hormone-secreting tumors have a high prevalence of rare headache phenotypes with or without autonomic features, suggesting that biochemical abnormalities within the hypothalamo-pituitary axis may play a role in headache. Somatostatin analogues may be highly effective at aborting headache associated with functionally active pituitary lesions, particularly in the case of acromegaly. A proposed mechanism for this is inhibition of nociceptive peptides. This article summarizes the clinical features, pathophysiology, and potential treatment approaches to pituitary tumor-associated headache. PMID- 18417029 TI - Oral manifestations associated with HIV infection. AB - Oral lesions are among the early signs of HIV infection and can predict progression to AIDS. The lesions commonly associated with the infection include oral candidiasis, herpes simplex infection, oral Kaposi's sarcoma, oral hairy leukoplakia, parotid gland enlargement, gingival diseases, xerostomia, and recurrent oral ulcerations. The introduction of highly active antiretroviral therapy has changed the epidemiology of some of the oral diseases associated with HIV infection. This review discusses the oral manifestations associated with HIV disease, the change in the pattern of the disease, and some research questions that need more emphasis from the research community. PMID- 18417030 TI - T-cell exhaustion in HIV infection. AB - Generation of memory T cells, which mediate immunity against microbes and cancers, relies, for optimal activity, on the interactions of multiple cell types that are highly regulated through the expression of soluble factors and negative and positive receptors. Their disruption will lead to aberrant immune responses, which can result in the invasion of the host by foreign pathogens. In chronic viral infections including HIV and hepatitis C virus, persistence of antigen and lack of CD4 help (HIV) disrupt memory T-cell function and induce defects in memory T-cell responses, which have been defined as T-cell exhaustion. In this review, we examine the molecular mechanisms involved in such T-cell dysfunction. Better understanding of these mechanisms will assist in the development of novel therapies to prevent the immune damage mediated by HIV infection. PMID- 18417031 TI - Regulatory T cells in HIV infection: who's suppressing what? AB - The role of regulatory T cells (Treg) in HIV pathogenesis is not fully elucidated. Persistent antigens, such as HIV, are believed to promote the expansion and activation of antigen-specific Treg, and several reports have described beneficial and detrimental roles for Treg in HIV pathogenesis. These apparently contradictory observations may arise from imprecision in enumerating Treg and the lack of definition of Treg subsets. New markers allowing more precise identification and purification of Treg for functional studies have been described recently, and these may open avenues for efficient isolation of pure, homogenous populations of human Treg. PMID- 18417032 TI - New players in cytokine control of HIV infection. AB - Cytokines are involved early in the pathogenesis of HIV infection and disease progression as a component of immunologic dysregulation and immunodeficiency and as determinants controlling virus replication. Several steps, before and after retroviral integration into host DNA in T cells and macrophages, are affected by cytokines whereas CCR5 and CXCR4 binding chemokines can interfere with viral entry. A growing number of potential players--including the gamma-common interleukin (IL)-7, IL-15, and IL-21 together with IL-17, IL-18, IL-19, IL-20, IL 23, and IL-27--are discussed in terms of their perturbation in HIV infection and of their effects on virus replication. Thus, an increasing intersection of HIV infection and the cytokine network represents a crucial determinant of virus replication and immunologic dysregulation and will likely play a key role in the development of effective strategies of HIV prevention and immunologic reconstitution. PMID- 18417033 TI - The potential of RNA interference-based therapies for viral infections. AB - RNA interference (RNAi) is a natural mechanism in cells that suppresses or silences the expression of aberrant or foreign genes. This activity is being developed as a potential antiviral therapeutic strategy. Studies in vitro, and some in vivo, appear to show the feasibility of using RNAi to treat virus infection. Therapeutic use of RNAi seems to be promising when directed against viruses that cause localized acute infections in accessible target cells. Therapeutic strategies using RNAi against viruses that cause chronic infections, such as HIV, hepatitis B virus, or hepatitis C virus, are more difficult to design, but studies have begun to address identifiable problems. Two clinical trials using RNAi have recently been initiated--one phase II trial against respiratory syncytial virus and a phase I trial against HIV. It will be of much interest to see whether nucleic acid therapies can offer another route to treating viral infection. PMID- 18417034 TI - Tat-regulated expression of RNA interference: triggers for the treatment of HIV infection. AB - HIV infection is a lifelong problem requiring continual medication for suppressing viral replication. Current strategies of antiretroviral drug combinations have proven effective in prolonging the time from infection to the symptoms of AIDS. Nevertheless, chemotherapy is not without its problems, which include toxicities and eventual emergence of virus mutants that are resistant to current antiretrovirals. Gene therapy refers to the introduction of effector oligonucleotides to transiently or stably alter gene expression or the delivery and expression of an exogenous gene within a specific target cell. A number of studies have demonstrated effective silencing/inhibition of HIV-1 replication by using RNA-based effector oligonucleotides for RNA interference. In this study, we have taken advantage of lentiviral vector-mediated delivery of anti-HIV short hairpin RNA for the treatment of HIV infection in hematopoietic cells. PMID- 18417035 TI - Role of natural killer cells in HIV pathogenesis. AB - Although the majority of research on immune cell recognition of HIV-infected cells has focused on CD8+ T cells with an eye towards vaccine development, innate immune recognition by natural killer (NK) cells has become a focus in recent years. Genetic and mechanistic data indicate that NK cells play a role during pathogenesis, and research on NK biology in the context of the broader immune response shows that NK cells are required to mount an effective antiviral response. HIV is able to escape cytotoxic T lymphocyte recognition by downmodulation of major histocompatibility complex class I receptors, which should enhance NK cytotoxicity against infected targets. However, the virus has evolved elaborate mechanisms to evade NK cell responses. Moreover, NK cell function as a whole is compromised through poorly understood mechanisms as a result of viremia. Further work on the role of NK cells during all stages of disease will further our understanding of the immune response against HIV. PMID- 18417036 TI - Treatment of hepatitis C genotype 2/3. PMID- 18417037 TI - What's new in the treatment of ascites and spontaneous bacterial peritonitis. AB - In recent years, there have been important advances in the clinical management of ascites and its related complications, such as hyponatremia, hepatorenal syndrome (HRS), and spontaneous bacterial peritonitis (SBP). Moreover, new drugs are currently being investigated for their potential usefulness in managing these complications. This article is not intended to comprehensively review all the literature published in recent years; rather, the authors discuss only studies they believe represent a potentially significant advance in this field. The following review is divided into two parts; the first discusses ascites and renal function abnormalities, including hyponatremia and HRS, and the second discusses SBP management. PMID- 18417038 TI - Pregnancy and liver disease. AB - Although liver disease does not occur frequently during pregnancy, when it does, it can cause significant morbidity and mortality for both fetus and mother. Recent advances in molecular genetics have provided some insight into the pathogenesis of diseases such as intrahepatic cholestasis of pregnancy (IHCP) and acute fatty liver of pregnancy. Newer studies linking serum bile acid levels with fetal outcome support the importance of this marker in IHCP. Randomized trials with ursodeoxycholic acid have shown promising results in reducing maternal symptoms and serum bile acid levels in IHCP. Although further studies must be performed, early strategies aimed at reducing viral loads in mothers may reduce perinatal transmission of hepatitis B. PMID- 18417039 TI - Hepatorenal syndrome: current management. AB - Hepatorenal syndrome (HRS) is a dreaded complication of advanced cirrhosis, with dismal prognosis if left untreated. Recent understanding of the pathophysiology underlying HRS development has led to many new treatments. Vasoconstrictors (eg, midodrine, terlipressin, or norepinephrine) together with albumin can improve the effective arterial blood volume with increased renal perfusion pressure. This combination has been proven successful in reversing HRS in approximately two thirds of patients. Transjugular intrahepatic portosystemic shunt (TIPS), which reduces portal pressure and decreases the sympathetic drive associated with portal hypertension, is effective in improving renal function. The combination of a TIPS and vasoconstrictor therapy has been shown to normalize renal function in a small number of patients. Liver transplantation corrects many of the pathophysiological abnormalities associated with HRS, and is the best treatment option. Patient outcome is improved further if the renal function is normalized prior to liver transplantation. PMID- 18417040 TI - Transjugular intrahepatic portosystemic shunt in the management of complications of portal hypertension. AB - Transjugular intrahepatic portosystemic shunt (TIPS) is a commonly used approach for managing many complications of portal hypertension. It is an attractive option due to its relative ease of creation (> 90% success rate) and the availability at most hospitals of an interventional radiologist capable of performing the procedure. TIPS is the preferred approach to control acutely bleeding esophageal or gastric varices that cannot be controlled with medical management. It is also now preferred to surgical shunts for preventing rebleeding in patients who rebleed despite adequate medical management. TIPS is more effective than large-volume paracentesis in controlling refractory cirrhotic ascites, with possibly a slight survival benefit but also increased encephalopathy. TIPS should be used to control refractory ascites in patients who cannot be managed with large-volume paracentesis. The role of TIPS in the treatment of hepatorenal syndrome is unclear; currently only patients with type 2 hepatorenal syndrome should be considered candidates for TIPS. PMID- 18417041 TI - Live donor liver transplantation: current status. AB - The inequality between supply of grafts and demand for transplants has forced the transplant community to devise ways to increase the number of available livers for transplant (ie, through use of extended criteria donor grafts and living donation). Since 2002, the number of live donor liver transplantations (LDLT) performed has declined due to concerns of donor safety and lack of clear outcome data establishing success equivalent to that of deceased donor liver transplantation (DDLT). Recent data suggest that LDLT outcomes are comparable with those of DDLT, provided a center has performed more than 20 procedures, both in patients with and without hepatitis C. Further studies are needed to define the optimal donor and the ideal recipient for LDLT. Results from a National Institutes of Health-funded consortium of nine transplant centers are highly anticipated. These data are expected to underscore the viability of LDLT as a life-saving therapy for certain patients with end-stage liver disease. PMID- 18417043 TI - Hepatitis C: current options for nonresponders to peginterferon and ribavirin. AB - Hepatitis C virus infection remains a significant health problem worldwide. The development of interferon-based therapies has led to increased success in eradicating this viral infection; however, no recent additions to the current treatment care regimen (peginterferon and ribavirin) have been made. Nonresponders to peginterferon/ribavirin make up a diverse patient population that can be difficult to manage. Through a better understanding of treatment response and viral kinetics, clinicians are employing altered dosing schedules to minimize the burden of viral illness. Consequently, clinicians can now manage nonresponders by providing supportive care and reserving retreatment for those most likely to respond to additional antiviral therapy, particularly when correctable factors from a previous treatment course with the potential for positive intervention have been identified. The decision for further medical management is based on clinical and laboratory parameters that have been shown to predict which patients will benefit most from another attempt at therapy. PMID- 18417042 TI - Diagnosis and management of cholangiocarcinoma. AB - Cholangiocarcinoma (CC) is a rare yet frequently fatal tumor that causes significant morbidity and mortality due to late presentation. Radiology is the mainstay of CC diagnosis; however, advances in understanding the pathogenesis of CC, including the role of oncogenes, inflammation-mediated genomic instability, and interleukin-6/STAT-3 signaling pathways, may allow development of new diagnostic and prognostic markers and targets for CC therapy. Although surgical resection is the standard of care for resectable CC, liver transplantation has shown excellent results in selected patients. The use of chemotherapy and radiotherapy are currently limited by marginal response rates, toxicity, and biliary complications. Locally ablative therapies in the form of transcatheter arterial chemoembolization and radioembolization are under investigation. Molecular therapies (eg, epidermal growth factor receptor, ErbB-2, and vascular endothelial growth factor receptor antagonists) and immunotherapy using diabodies are also under investigation for treatment of unresectable CC. PMID- 18417044 TI - Early detection of liver cancer: diagnosis and management. AB - Hepatocellular carcinoma (HCC) is the fifth most common cause of cancer deaths worldwide, and the incidence is rising. Despite a wide array of treatment options, fewer than half of candidates for potentially curative treatments receive them. The diagnosis and management of HCC require a multidisciplinary approach involving various clinical specialties. The foundation of diagnosis is high-quality imaging, with MRI being the test of choice. Some patients also require guided biopsy when MRI is equivocal. Treatment options depend upon the tumor stage and the degree of underlying synthetic dysfunction. Potentially curative treatments include surgical resection and transplantation. Other treatments that prolong survival include percutaneous ablation and transarterial chemoembolization. A new oral agent, sorafenib, was recently shown to prolong survival in patients with advanced HCC. By increasing surveillance and treatment of HCC, outcomes for these patients may be improved. PMID- 18417045 TI - The obesity epidemic and nonalcoholic fatty liver disease in children. AB - Childhood obesity is a worldwide health problem associated with an increase in the prevalence and severity of nonalcoholic fatty liver disease (NAFLD). This review covers the progress made between 2005 and 2007 in understanding the epidemiology, histology, and treatment of pediatric NAFLD. The number of children with NAFLD presents a major public health crisis. Noninvasive diagnostic tools offer future promise, but currently are unable to grade and stage disease. Therefore, pediatric NAFLD remains a clinico-pathological diagnosis requiring direct demonstration of liver steatosis and the exclusion of other causes of fatty liver and/or hepatitis. There are currently no proven therapies for NAFLD in children; however, TONIC (Treatment of Nonalcoholic Fatty Liver Disease in Children), the first multicenter clinical trial of pediatric NAFLD, is currently in progress. Such studies are imperative to address fundamental questions regarding cause and cure. PMID- 18417046 TI - Nonalcoholic fatty liver disease as a component of the metabolic syndrome. AB - Nonalcoholic fatty liver disease (NAFLD) is an important cause of liver-related morbidity and mortality. The association between NAFLD and the metabolic syndrome is well established: the presence of the metabolic syndrome signifies advanced histology in NAFLD patients. Emerging data indicate that patients with NAFLD have a significantly higher prevalence of cardiovascular disease. This article reviews the definitions of the two syndromes, their association, and the cardiovascular risk conferred by both when they occur separately and together. This review also discusses management options for the syndromes, including treating the individual components of the metabolic syndrome in NAFLD patients. PMID- 18417048 TI - Asthma-obesity relationship: association or causality? PMID- 18417047 TI - Novel therapies in hepatitis B and C. AB - Chronic hepatitis B and C affect approximately 500 million people in the world, with substantial disease burden including liver cirrhosis and hepatocellular carcinoma. For chronic hepatitis B, two treatment strategies are currently available, both with suboptimal response and significant side effects. Promising new drugs are approaching the stage of approval; however, these agents still need further development to control this disease. Based on the understanding of the hepatitis C virus life cycle, new treatment developments for chronic hepatitis C tend to succeed rapidly; therefore, it is only a matter of time before new therapies emerge. This review summarizes the most important new agents available for treatment of chronic hepatitis B and C. PMID- 18417050 TI - The mold conundrum in chronic rhinosinusitis: where do we stand today? AB - Chronic rhinosinusitis (CRS) is an inflammatory disorder affecting the nose and paranasal sinuses. Although bacteria have long been implicated as pathogens in most forms of CRS, fungi may be responsible for some forms. Several recent studies demonstrated that, under optimal conditions, fungi can be identified in the nose and paranasal sinuses of nearly every individual (including all CRS patients). An aberrant immune response to these ubiquitous fungi has been suggested to explain the development of CRS in some individuals. Several mechanisms requiring additional research, including adequate controls, have been proposed and are reviewed in this article. Although preliminary trials suggested that CRS signs and symptoms improve upon treatment with topical and oral antifungal agents, several double-blind, placebo-controlled trials demonstrated the contrary. In the absence of convincing immunologic data and evidence of clinical improvement upon therapy with antifungal agents, the case against fungi remains unproven. PMID- 18417051 TI - Sublingual immunotherapy and allergic rhinitis. AB - This paper reviews the safety and efficacy of sublingual immunotherapy (SLIT) in the treatment of allergic rhinitis. The literature from 1986 through 2007 shows approximately a 6000-fold range in doses found to be effective with SLIT. However, recent studies in large patient populations have demonstrated a clear dose response with an effective dose range that appears to be equivalent to one to two times the monthly subcutaneous immunotherapy dose administered daily or weekly (ie, 15 to 30 microg of major allergen). Further study is needed to establish the optimal dose and dosing schedule for each formulation. Local reactions (eg, oral itchiness) are common, and serious adverse reactions, although rare, have been reported. Cost-effective analysis cannot be made until the effective dose is established. SLIT appears to be a promising treatment for allergic rhinitis, but it is currently considered investigational in the United States until a formulation approved by the US Food and Drug Administration is available. PMID- 18417052 TI - Upper airways reactions to cold air. AB - Cold air-induced rhinitis is a common complaint of individuals with chronic allergic or nonallergic rhinitis and those with no chronic nasal disease. It is characterized by rhinorrhea, nasal congestion, and nasal burning that appear within minutes of exposure to cold air and dissipate soon after exposure is terminated. The symptoms of cold-air rhinitis are reproduced experimentally with nasal cold-air provocation. This procedure has shown that nasal mast cell activation and sensory nerve stimulation are associated with the development of nasal symptoms. Sensory nerve activation generates a cholinergic reflex that leads to rhinorrhea; therefore, anticholinergic agents are highly effective in treating cold-air rhinitis. Experimental data suggest that individuals with nasal cold-air sensitivity may have reduced ability to compensate for the water loss that occurs during exposure to cold air. Therefore, the symptoms of cold air induced rhinitis may reflect the activation of compensatory mechanisms to restore mucosal homeostasis. PMID- 18417053 TI - Complementary therapy in allergic rhinitis. AB - The term complementary/alternative medicine (CAM) refers to those therapeutic and diagnostic approaches different from conventional allopathic medicine. CAM may encompass homeopathy, acupuncture, phytotherapy, antioxidant therapy, and numerous holistic or behavioral techniques. Allergists and physicians of all disciplines are confronted with patients using CAM treatments, making it imperative that they become familiar with the scientific literature surrounding them. Given the high prevalence of allergic diseases and associated costs of CAM treatments, proof of CAM therapies is needed to establish appropriate guidelines for their use. Efficacy of CAM modalities should be established with randomized, double-blind, placebo-controlled trials, including adverse-effects monitoring. Of all the CAM therapies examined to treat allergic rhinitis, some herbal therapies and antioxidants demonstrate a trend toward some clinical efficacy. Researchers have yet to determine how to integrate these CAM modalities into the general treatment paradigm of allergic rhinitis. PMID- 18417054 TI - Allergen avoidance. AB - The systematic avoidance of indoor allergens by modification of houses, furnishings, or hygiene practices has long been advocated to reduce both the incidence of allergic diseases in at-risk infants and exacerbations or symptoms in those previously sensitized with such a disease. However, such advocacy is now under challenge, due to both a lack of evidence of clinical efficacy of avoidance measures and rapidly changing models of disease causation and pathology. This article reviews the current dilemmas in understanding the complexity of allergen sources, disease risks, nature of bioaerosol exposure, and the inadequacies of many available methods. PMID- 18417055 TI - Intranasal corticosteroids in the treatment of acute rhinosinusitis. AB - Acute rhinosinusitis (ARS) is one of the most frequent diagnoses in clinical practice. Common symptoms include anterior and/or posterior nasal drainage, nasal obstruction, and facial pain, pressure, or sensation of fullness. ARS etiology is more often viral than bacterial, and infection-initiated inflammation is the primary pathophysiologic mechanism responsible for symptoms. Because ARS is usually a self-limited process, the primary goal of treatment is symptom relief, which can be more effectively achieved by targeting the underlying inflammation. Intranasal steroids (INS) have been shown to reduce components of respiratory inflammation and increase symptom relief for ARS when used alone or combined with antibiotics. Based on current evidence in support of INS therapy, the European Position Paper on Rhinosinusitis and Nasal Polyps 2007 guidelines and a recent Cochrane meta-analysis recommend using INS in the treatment of ARS, both as monotherapy and as an adjunct to antibiotics. PMID- 18417056 TI - The effect of air pollution on asthma and allergy. AB - Air pollution exposure is associated with increased asthma and allergy morbidity and is a suspected contributor to the increasing prevalence of allergic conditions. Observational studies continue to strengthen the association between air pollution and allergic respiratory disease, whereas recent mechanistic studies have defined the prominent role of oxidative stress in the proallergic immunologic effects of particulate and gaseous pollutants. The identification of common genetic polymorphisms in key cytoprotective responses to oxidative stress has highlighted the importance of individual host susceptibility to pollutant induced inflammation. Future therapy to reduce the adverse effects of air pollution on allergic respiratory disease will likely depend on targeting susceptible populations for treatment that reduces oxidative stress, potentially through enhancement of phase 2 enzymes or other antioxidant defenses. PMID- 18417057 TI - Nasal reflexes: implications for exercise, breathing, and sex. AB - Nasal patency, with both congestion and decongestion, is affected in a wide variety of reflexes. Stimuli leading to nasal reflexes include exercise; alterations of body position, pressure, and temperature; neurologic syndromes; and dentistry. As anticipated, the vagal and trigeminal systems are closely integrated through nasobronchial and bronchonasal reflexes. However, perhaps of greater pathophysiologic importance are the naso-hypopharyngeal-laryngeal reflexes that become aggravated during sinusitis. None other than Sigmund Freud saw deeply beyond the facial adornment and recognized the deeper sexual tensions that can regulate nasal functions and psychoanalytical status. Wine, women, and song are linked with airflow through the nose-the nose, which by any other name would still smell as sweetly. PMID- 18417058 TI - Anti-inflammatory treatment of chronic rhinosinusitis: a shifting paradigm. AB - Treatment of chronic rhinosinusitis still represents an unmet medical need. Presently, the US Food and Drug Administration has not approved any drugs for the treatment of this common condition. Various inflammatory processes are involved in the pathogenesis of chronic rhinosinusitis. Treatment of infection and surgical intervention to correct anatomic abnormalities often are inadequate in management when singly employed. Anti-inflammatory therapy such as topical cortico-steroids and more recently long-term, low-dose macrolide therapy has been integrated into the treatment algorithm for chronic rhinosinusitis. Better classification and point-of-care identification of inflammatory features may improve choice of anti-inflammatory therapy and thus outcome. PMID- 18417059 TI - Recent advances in the role of NKT cells in allergic diseases and asthma. AB - Asthma is the result of chronic airway inflammation that is dominated by the presence of eosinophils and CD4(+) T lymphocytes. CD4(+) T cells include several subsets and play a critical role in orchestrating the inflammation, predominantly by secreting interleukin-4 and interleukin-13. Recently, research identified a new subset of T cells, natural killer T (NKT) cells, which also express the CD4 marker. In contrast to conventional CD4(+) T cells, NKT cells do not respond to peptide antigens, but rather to glycolipids. In animal models of asthma, direct activation of NKT cells by glycolipids results in the secretion of extensive amounts of cytokines and triggers the development of airway hyperreactivity. Moreover, in patients with chronic asthma, NKT cells can be found in bronchoalveolar lavage fluids in significant amounts. These data strongly suggest that NKT cells play an important role in asthma pathogenesis. PMID- 18417060 TI - Steroid resistance in asthma: mechanisms and treatment options. AB - Glucocorticoid insensitivity presents a profound management problem in patients with asthma because conventional therapies are not effective. Glucocorticoids, acting through the glucocorticoid receptor (GR), are able to selectively repress inflammatory gene expression by utilizing several distinct mechanisms targeting nuclear factor-varphiB and activator protein-1 activation complexes and by effects on mitogen-activated protein kinases. Different model systems often activate distinct sets of signaling molecules and different glucocorticoid responsiveness may result from differences in concentrations and timing of steroid treatment of cells, GR expression levels, and the precise inflammatory stimulus used. Thus, abnormal activation of many signaling pathways may affect corticosteroid responsiveness in patients with corticosteroid-resistant asthma. Understanding the molecular mechanisms of GR action and inaction may lead to the development of new anti-inflammatory drugs or enable clinicians to reverse the relative steroid-insensitivity that is characteristic of some patients with severe asthma. PMID- 18417061 TI - Will CETP inhibition survive the demise of torcetrapib? PMID- 18417062 TI - Optimal medical therapy with or without percutaneous coronary intervention for stable coronary disease: the COURAGE Study. PMID- 18417063 TI - Triglycerides and the risk of coronary artery disease. PMID- 18417064 TI - Prevention and noninvasive management of coronary atherosclerosis in patients with diabetes. AB - Diabetes mellitus (DM) is a worldwide epidemic. Its prevalence is rapidly increasing in both developing and developed countries. Coronary heart disease (CHD) is highly prevalent and is the major cause of morbidity and mortality in patients with diabetes. Individuals with prediabetes states, with or without known CHD, should undergo lifestyle modifications aimed at preventing DM. In patients with CHD and DM, routine use of aspirin and an angiotensin-converting enzyme inhibitor, along with strict glycemic, blood pressure, and lipid control, is strongly recommended. Intense insulin therapy may be needed for glycemic control, and high-dose statin therapy may be needed for lipid control. For blood pressure control, angiotensin-converting enzyme inhibitors and angiotensin receptor blockers are considered first-line therapy. Noncompliance with medications and/or lifestyle measures and underprescription of evidence-based therapies remain important unsolved problems. PMID- 18417065 TI - Alcohol consumption and heart failure: a systematic review. AB - Heart failure (HF) remains a major public health issue. It is estimated that about 500,000 Americans per year are diagnosed with HF. Despite advanced medical and surgical treatments for HF, mortality after the onset of HF is still high, thereby underscoring the importance of primary prevention. Among modifiable lifestyle factors, alcohol consumption appears to play a role in the development of HF. Although excessive drinking has been known to lead to alcoholic cardiomyopathy and light-to-moderate drinking may confer some cardiovascular benefits, recent studies suggest it is not only the quantity, but also drinking patterns and genetic factors, that may influence the relation between alcohol consumption and cardiovascular disease. This article reviews current evidence on the association between alcohol consumption and HF. PMID- 18417066 TI - Racial and ethnic differences in hypertension. AB - Hypertension is a major cause of disease burden in all racial and ethnic groups and in both developing and developed regions and countries. Much of the racial and ethnic disparity in cardiovascular outcomes can be attributed to the excess burden of hypertension. Racial and ethnic differences in blood pressure occur because of biology and sociology. Causes of racial differences in blood pressure likely begin early in life and reflect the complex relationship of these gene and environment interactions. Hypertension treatment and control remain less than optimal worldwide, and awareness is still a problem in many racial and ethnic groups. Instituting lifestyle changes for the primary prevention and treatment of hypertension among the general population would decrease prevalence and be effective in eliminating many racial and ethnic differences. This review highlights racial and ethnic differences in the prevalence and incidence of hypertension and identifies contributing factors associated with these differences. PMID- 18417067 TI - Rheumatoid arthritis and cardiovascular disease. AB - Rheumatoid arthritis (RA) is a chronic systemic inflammatory disease affecting approximately 1% of the adult general population. Cardiovascular disease is recognized as the leading cause of death in RA patients, accounting for nearly 40% of their mortality. Patients with RA are at a twofold increased risk for myocardial infarction and stroke, with risk increasing to nearly threefold in patients who have had the disease for 10 years or more. Congestive heart failure appears to be a greater contributor to excess mortality than ischemia. This increased cardiovascular disease risk in RA patients seems to be independent of traditional cardiovascular risk factors. Pathogenic mechanisms include pro oxidative dyslipidemia, insulin resistance, prothrombotic state, hyperhomocysteinemia, and immune mechanisms such as T-cell activation that subsequently lead to endothelial dysfunction, a decrease in endothelial progenitor cells, and arterial stiffness, which are the congeners of accelerated atherosclerosis observed in RA patients. This paper discusses pathogenic mechanisms, effects of methotrexate, tumor necrosis factor antagonists, steroids, and statins, with a perspective on therapy. PMID- 18417068 TI - PPARgamma agonists and coronary atherosclerosis. AB - The prevalence of type 2 diabetes mellitus (T2DM) is growing at an alarming rate and reaching epidemic proportions, and cardiovascular disease continues to be one of the leading causes of death in the United States. The key relationship between these two diseases (knowing that T2DM is a strong risk factor for cardiovascular disease) is insulin resistance and the detrimental effect it has on macrovasculature. Thiazolidinediones (TZDs) are peroxisome proliferator-activated receptor gammaagonists that are beneficial in the treatment of T2DM and have the added benefit of modifying lipid profiles. This review discusses the basic science linking insulin resistance to atherosclerosis and describes the major TZD trials in the recent literature. It also addresses the clinical implications of these studies and media scrutiny surrounding the recent controversial report that TZDs may be linked to an increased risk of myocardial infarction. PMID- 18417069 TI - Adjunctive interventions in myocardial infarction: the role of statin therapy. AB - Statin therapy has reduced cardiovascular morbidity and mortality across the spectrum of atherosclerosis. The administration of statins has been demonstrated to be effective in primary and secondary prevention clinical trials evaluating patients with high and low risk-factor profiles. The presumed mechanism of benefit of hypolipidemic therapy in the prevention of atherosclerotic disease was a reduction in the deposition of atherogenic lipoproteins in vulnerable areas of the coronary vasculature. Subsequent experimental studies with statins demonstrated a variety of potentially beneficial effects that would extend clinical benefit beyond lipid-lowering per se. Statin therapy beneficially alters inflammation, coagulation and fibrinolytic parameters, endothelial function, vasoreactivity, and platelet function. The demonstration of the non-lipid or pleiotropic effects provided the theoretical basis for a possible role as an adjunctive therapy in acute coronary syndromes. Retrospective analysis of a variety of early trials indicated a potential benefit of statins during acute ischemic syndromes. Recent clinical trials have addressed this important clinical question in a prospective controlled manner. The Myocardial Ischemia Reduction with Aggressive Cholesterol Lowering (MIRACL) and the Thrombolysis In Myocardial Infarction (TIMI)-22 studies present strong clinical evidence in favor of the administration of statins as adjunctive therapy in acute ischemic syndromes. PMID- 18417070 TI - Aspirin resistance in atherosclerosis. AB - Clinically, aspirin resistance is defined as the failure of aspirin therapy to prevent an acute vascular thrombotic event despite regular intake of appropriate doses. In the laboratory, aspirin resistance encompasses the drug's failure to attain a particular level of platelet inhibition. From a clinical standpoint, the inability of aspirin to prevent a thrombotic event, despite appropriate cyclooxygenase-1 inhibition, implies the involvement of other factors. Evidence is emerging that aspirin resistance, as defined by residual platelet activity, merely reflects an individual's enhanced basal platelet function and suggests a hereditary component. Due to the multifactorial nature of cardiovascular disease, it is likely that a single therapy like aspirin cannot fully treat and prevent all thrombotic complications in the setting of atherosclerosis. PMID- 18417071 TI - Intensive statin therapy in acute coronary syndromes. AB - Acute coronary syndromes (ACS) represent an enormous disease burden, especially in the Western world, where they are one of the largest causes of mortality and morbidity. Patients suffering an acute coronary event are at very high risk of further coronary events, and although improvements in medical therapy over the past two decades have significantly reduced the risk, it remains high. 3-Hydroxy 3-methylglutaryl coenzyme A reductase inhibitors (statins) are now fully established in the primary and secondary prevention of stable coronary heart disease, and recently a substantial body of evidence has proven their efficacy in ACS. This paper examines the evidence for statin use in ACS. PMID- 18417073 TI - Proatherogenic high-density lipoprotein, vascular inflammation, and mimetic peptides. AB - Atherosclerosis is an example of an inflammatory disorder. During the acute phase and under inflammatory conditions, high-density lipoprotein (HDL), which is normally anti-inflammatory, can become proinflammatory. Reactive oxygen species generated by several enzyme systems can modify phospholipids and sterols, producing oxidized phospholipids and oxidized sterols that reduce the capacity of HDL to protect against undesirable oxidative modifications of molecules. In animal models of dyslipidemia, diabetes, vascular inflammation, and chronic rejection, it is observed that reducing oxidative and inflammatory pressure will help HDL regain its protective role. One way to accomplish this is through the use of apolipoprotein A-I mimetic peptides, which remove oxidation products from lipoproteins and cell membranes, returning normal structure and function to low density lipoprotein and HDL. These mimetic peptides markedly reduce atherosclerosis in animal models. Published studies of apolipoprotein mimetic peptides in models of inflammatory disorders other than atherosclerosis suggest that they have efficacy in a wide range of inflammatory conditions. PMID- 18417072 TI - Novel markers of inflammation in atherosclerosis. AB - Inflammation plays a key role in the pathogenesis of atherosclerosis. Understanding the process of inflammation as it pertains to atherosclerosis has provided researchers with multiple opportunities to identify novel markers for use in cardiovascular disease management. This article discusses the inflammatory cascade as it pertains to atherosclerosis and some of the well-studied markers of inflammation. It also discusses the limitations of current risk stratification models and characteristics of a good biomarker. PMID- 18417074 TI - Recombinant aequorin with a reactive cysteine residue for conjugation with maleimide-activated antibody. AB - The mutated recombinant aequorin with a reactive cysteine residue (Cys-aequorin) was highly purified and then conjugated with a maleimide-activated antibody without significant loss of luminescence activity. The conjugate ratio of Cys aequorin to heavy chain of immunoglobulin G (IgG) was estimated to be 1:1. To test the bioluminescent immunoassay with aequorin-labeled antibody, alpha fetoprotein (AFP), a serological marker of liver cancer, was used as a model analyte. The measurable range of AFP was 0.02 to 200 ng/ml with the coefficient of variation between 2.1 and 4.5%. PMID- 18417075 TI - Fluorescence assay for mitochondrial permeability transition in cardiomyocytes cultured in a microtiter plate. AB - Mitochondrial permeability transition pore (MPTP) is a voltage-dependent, large conductance channel of the inner mitochondrial membrane with an important role in a range of pathophysiological conditions. To facilitate studies of pharmacological pore modulation, we describe an assay in a model using neonatal cardiomyocytes in a 96-well microtiter plate format. In the presence of mitochondrial membrane potential Delta Psi m, accumulation of rhodamine-123 in mitochondria (40,000 cells/well, 2.6 microM rhodamine-123) caused fluorescence signal quenching. Following substitution of dye-free buffer, dequenching occurred on the distribution of rhodamine-123 into the extracellular volume. The addition of a small buffer volume containing digitonin (final concentration 10 microg/ml) and Ca(2+) (final concentrations up to 100 microM free Ca(2+)) caused dequenching (Delta F) due to Delta Psi m dissipation by MPTP, as evidenced by inhibition in the presence of cyclosporin A (0.2-2 microM) and facilitation by pH 6.2. Delta F due to Delta Psi m-dissipating agent carbonyl cyanide p trifluoromethoxyphenylhydrazone (FCCP) or alamethicin (10 microM) was insensitive to either pH or cyclosporin A. Inhibition of Ca(2+)-induced (but not of FCCP- or alamethicin-induced) Delta F by glycogen synthase kinase 3beta (GSK3 beta) antagonist SB216763 and adenosine, acting at the level of intracellular signaling and plasma membrane receptors, respectively, is shown to illustrate potential applications of this assay. Limitation of the assay to cells with energized mitochondria is stressed. PMID- 18417076 TI - Molecular basis for the ATPase activity of CFTR. AB - CFTR is a member of the ABC (ATP binding cassette) superfamily of transporters. It is a multidomain membrane protein, which utilizes ATP to regulate the flux of its substrate through the membrane. CFTR is distinct in that it functions as a channel and it possesses a unique regulatory R domain. There has been significant progress in understanding the molecular basis for CFTR activity as an ATPase. The dimeric complex of NBD structures seen in prokaryotic ABC transporters, together with the structure of an isolated CF-NBD1, provide a unifying molecular template to model the structural basis for the ATPase activity of CFTR. The dynamic nature of the interaction between the NBDs and the R domain has been revealed in NMR studies. On the other hand, understanding the mechanisms mediating the transmission of information from the cytosolic domains to the membrane and the channel gate of CFTR remains a central challenge. PMID- 18417077 TI - Erythropoietin has an antiapoptotic effect after myocardial infarction and stimulates in vitro aortic ring sprouting. AB - Aims were to explore if darbepoietin-alpha in mouse can induce angiogenesis and if moderate doses after myocardial infarction stimulates periinfarct capillary and arteriolar densities, cell proliferation, and apoptosis. Myocardial infarction was induced by ligation of LAD. Mouse aortic rings (0.8mm) were cultured in matrigel and the angiogenic sprouting was studied after addition of darbepoietin-alpha with and without VEGF-165. After 12 days the hemoglobin concentration was 25% higher in the darbepoietin-alpha treated mice than in the control group. No difference in capillary densities in the periinfarct or noninfarcted areas was seen with darbepoietin-alpha. Cell proliferation was about 10 times higher in the periinfarct area than in the noninfarcted wall. Darbepoietin-alpha treatment led to a decrease of cell proliferation (BrdU, (p<0.02)) and apoptosis (TUNEL, p<0.005) with about 30% in the periinfarct area. Darbepoietin-alpha and VEGF-165 both independently induced sprouting from aortic rings. The results suggest that darbepoietin-alpha can induce angiogenesis but that moderate doses after myocardial infarction are not angiogenic but antiapoptotic. PMID- 18417078 TI - Examination and expansion of the substrate range of m-hydroxybenzoate hydroxylase. AB - The gene encoding m-hydroxybenzoate hydroxylase (mobA) was cloned from Comamonas testosteroni GZ39. MobA converts m-hydroxybenzoate and to a lesser extent p hydroxybenzoate to protocatechuate. To explore the structural and functional relationships in phenolic acid monooxygenases, MobA was subjected to in vitro mutagenesis by error-prone PCR and the mutant MobAs were screened for their ability to oxidize phenol or 3-aminophenol. A mutant MobA with a single V257A substitution was able to transform phenol to catechol, providing the first example of monooxygenase acting on phenolic acids that can also hydroxylate phenol. The mutant MobA also has enhanced ability to transform resorcinol, hydroquinone, p-hydroxybenzoate, 2,5-dihydroxybenzoate, 3,4-dihydroxybenzoate, 3 chlorophenol, 4-chlorophenol, 4-chlororesorcinol, and 4-nitrophenol. Several MobA mutants were obtained for their ability to transform 3-aminophenol to a related substituted catechol. Mutant MobAs with single amino acid substitutions (H135P, A400G, or D416A) were derived from these mutants and verified for their ability to transform 3-aminophenol. PMID- 18417079 TI - Glucuronic acid can extend O-linked core 1 glycans, but it contributes only weakly to the negative surface charge of Drosophila melanogaster Schneider-2 cells. AB - Previous studies of the mucin-type O-glycome of the fruit fly Drosophila melanogaster have revealed a restricted pattern of neutral core-type glycans corresponding to the Tn-(GalNAcalpha) and the T-antigen (Galbeta1-3GalNAcalpha). In particular, no extension of the core 1 glycan with acidic sugars, like sialic acid, was detected. Here we report on the identification of an acidic O-linked trisaccharide expressed on secreted endogenous and recombinant glycoproteins of the embryonal hemocyte-like Drosophila Schneider-2 (S2) cell line. The glycan is composed of glucuronic acid, galactose and N-acetylgalactosamine and its structure was determined as GlcA1-3Gal1-3GalNAc. The O-linked trisaccharide resembles the peripheral structures of acidic D. melanogaster glycosphingolipids. Glucuronic acid may substitute for sialic acid in this organism, however its expression on the S2 cell surface may only marginally contribute to the negative surface charge as revealed by free-flow cell electrophoresis prior to and after beta-glucuronidase treatment of the cells. PMID- 18417080 TI - Suggesting childhood food illness results in reduced eating behavior. AB - Previous studies have shown that suggesting childhood events can influence current self-reported attitudes towards future behavior. This study shows that suggesting a false past event (i.e. becoming sick on a specific food during childhood) can modify present behavior (i.e. reduce eating of the food). Participants screened to be normal eaters received or did not receive a suggestion that they likely became sick on spoiled peach yogurt as a child. One week later they took part in an allegedly separate marketing taste-test study, during which they rated preferences for a variety of crackers and yogurts. After completing ratings, participants were invited to freely eat the remaining food while completing questionnaires. Results revealed that the participants receiving the suggestion expressed lower preference specifically for peach yogurt, and ate less yogurt of all the types, while not differing in eating of crackers. These results demonstrate that suggesting false past events influences subsequent behavior. PMID- 18417081 TI - A simple and safe anastomosis in pancreaticogastrostomy using mattress sutures. AB - When a suture does not trap a sufficient volume of pancreatic parenchyma in the conventional pancreatic anastomosis, laceration may be caused by even modest tension. Therefore, we used mattress sutures with gastric wall, which works protectively. An opening was made on the posterior wall of the stomach. By using a 3-0 polypropylene monofilament thread with a straightened needle at each end, the proximal gastric wall, the pancreas, and the distal gastric wall were sutured sequentially to accomplish anastomosis in a U-like fashion. After 2 to 4 additional sutures were completed, the pancreatic stump was invaginated into the stomach and all sutures are tied. Mattress sutures were made on the pancreas, which was held between the superior and inferior walls of the stomach. Postoperative amylase levels in the drainage fluid decreased significantly and none of the 17 patients developed pancreatic fistula formation. The technique is simple and shortens the time required for anastomosis. PMID- 18417082 TI - The role of surgery in the treatment of recurrent gastric cancer. AB - BACKGROUND: The purpose of the current study was to determine the role of surgery in the treatment of recurrent gastric cancer. METHODS: Of the 347 patients with recurrent gastric cancer, 61 patients (17.8%) who underwent surgery were evaluated retrospectively. The underlying causes and types of surgery, survival, and postoperative quality of life were analyzed. RESULTS: The most common cause of surgery was intestinal obstruction due to carcinomatosis. Complete resection was possible in 15 patients (24.6 %), including 10 gastric remnant recurrences, and 2 hepatic and 3 ovarian metastases. The survival of patients who had complete resection was significantly longer than the other groups (52.2 months for complete resections, 13.1 months for palliative procedures, and 8.7 months for laparotomy alone, respectively) (P < .05). The median hospital-free survival (HFS) durations were 9.4, 2.9, and 2.2 months for incomplete resection, bypass/enterostomy, and laparotomy only, respectively (P < .05). CONCLUSION: Surgical treatment in recurrent gastric cancer is rarely indicated; however, if complete resection could be accomplished, long-term survival can be expected. Bypass surgery for symptom palliation did not increase the HFS. PMID- 18417083 TI - The use of radiation therapy after breast-conserving surgery in hormonally treated breast cancer patients is dependent on patient age, geographic region, and surgeon specialty. AB - BACKGROUND: The use of radiation therapy (RT) after breast-conserving surgery (BCS) is inconsistent in patients treated with hormonal therapy (HT). We sought to identify factors influencing the decision to use RT in this setting. METHODS: Patients in the North American Fareston vs. Tamoxifen Adjuvant (NAFTA) trial who had BCS were evaluated for factors influencing the use of RT using univariate and multivariate analyses. RESULTS: Of the 1,811 patients enrolled in the NAFTA trial, 1,222 (67.4%) had BCS. Of these, 241 (19.7%) did not have RT. There were no significant differences in tumor grade, lymphovascular invasion, estrogen receptor status, or nodal status between those who received RT and those who did not. On univariate analyses, patients who did not receive RT were more likely to be older (median 72 versus 66 years, P < .001), have larger tumors (median 1.35 versus 1.10 cm, P = .009), and be progesterone negative (18.3% versus 13.1%, P = .048). Surgeons in the West were most likely to omit RT, whereas those in the Midwest were least likely to omit it (26.7% versus 7.2%, P < .001). Surgical oncologists were more likely to omit RT after BCS than general surgeons (21.8% versus 13.7%, P < .001). Academic affiliation (P < .001), number of breast procedures performed per year (P = .017), and the percentage of breast practice (P = .019) also correlated with whether or not RT was used. On multivariate analysis, patient age (P < .001), geographic region (P = .006), and surgeon specialty (P = .027) remained significant. CONCLUSION: Patient age, geographic region, and surgeon training influence the decision to use RT after BCS in patients receiving HT. PMID- 18417084 TI - Learning style and laparoscopic experience in psychomotor skill performance using a virtual reality surgical simulator. AB - BACKGROUND: People learn in different ways, and training techniques and technologies should accommodate individual learning needs. This pilot study looks at the relationship between learning style, as measured with the Multiple Intelligences Developmental Assessment Scales (MIDAS), laparoscopic surgery experience and psychomotor skill performance using the MIST VR surgical simulator. METHODS: Five groups of volunteer subjects were selected from undergraduate tertiary students, medical students, novice surgical trainees, advanced surgical trainees and experienced laparoscopic surgeons. Each group was administered the MIDAS followed by two simulated surgical tasks on the MIST VR simulator. RESULTS: There was a striking homogeny of learning styles amongst experienced laparoscopic surgeons. Significant differences in the distribution of primary learning styles were found (P < .01) between subjects with minimal surgical training and those with considerable experience. A bodily-kinesthetic learning style, irrespective of experience, was associated with the best performance of the laparoscopic tasks. CONCLUSION: This is the first study to highlight the relationship between learning style, psychomotor skill and laparoscopic surgical experience with implications for surgeon selection, training and credentialling. PMID- 18417085 TI - Adenocarcinoma of the gastroesophageal junction after bariatric surgery. AB - BACKGROUND: The development of upper gastrointestinal malignancies after bariatric surgery has not been well characterized. Our objective was to review the experience of patients with distal esophageal cancer that was diagnosed after bariatric surgery. METHODS: A retrospective review was conducted to identify patients who had undergone bariatric surgery (1999 to 2006) and who later developed high-grade dysplasia or adenocarcinoma of the distal esophagus. RESULTS: Three patients (of 2,875 [0.1%]) developed esophageal cancer: 2 after Roux-en-Y gastric bypass and 1 after vertical banded gastroplasty. All three patients had complaints of reflux, and two were treated with esophagectomy. The third patient presented with invasive carcinoma and died 2 years after diagnosis. CONCLUSIONS: Our findings emphasize the importance of precise endoscopic evaluation before bariatric surgery in patients with gastroesophageal reflux disease (GERD), of the necessity for continuing postsurgical surveillance in patients with known Barrett's esophagitis, and of early evaluation in patients who develop new symptoms of GERD after bariatric surgery. PMID- 18417086 TI - Construct validity of an objective assessment method for laparoscopic intracorporeal suturing and knot tying. AB - BACKGROUND: The ideal objective assessment method for laparoscopic technical skills is difficult to achieve in the operating room. Recent "VR2OR" studies have used a blinded, 2-reviewer error-based video tape analysis for intraoperative performance assessment. This study examines the validity of this methodology applied to laparoscopic intracorporeal suturing and knot tying. METHODS: Four groups of subjects--experts (EX), surgery residents trained to expert criterion levels using simulation (TR), surgery residents receiving no supplemental training (NR), and medical students receiving simulation-based training (MS)- performed the fundal suturing portion of a laparoscopic Nissen fundoplication and were video-recorded for analysis. Two separate groups of surgeon reviewers (K.V.S. + M.B.; I.-P.H. + A.G.) were trained to evaluate laparoscopic suturing and knot tying performance using specific metrics. Subjects' operative performance was assessed by reviewers blinded to their training status and scored using an error-based, step specific scoring system to an inter-rater agreement of 80% or greater. Three primary performance measures were assessed: time, errors, and needle manipulations and comparisons between groups were made using a 1-way analysis of variance (ANOVA) with post-test. RESULTS: A total of 40 fundal sutures (10 in each group) were scored by 2 separate rater groups with inter rater agreement consistently greater than 80%. Inter-rater agreement was highest with the EX group (91%, range 76%-100%) and lowest with the NR group (85%, range 81%-98%). On average, the EX group significantly outperformed the other groups with regards to time (P <.0001), errors (P <.002), and needle manipulations (P <.01). Performance of the TR group was comparable to the EX group with regards to errors and manipulations (P = not significant [NS]), and outperformed the NR and MS groups with regards to time (P <.05 and P <.001). Performance between the NR and MS groups were similar for all 3 measures. CONCLUSIONS: This assessment method demonstrates discriminative validity. Time appears to be the most sensitive indicator of skill level, as significant differences between EX, TR, and NR/MS groups were seen. The methodology is transferable across different reviewers and is acceptable for high-stakes assessment. PMID- 18417087 TI - Postoperative antibiotic therapy for children with perforated appendicitis. PMID- 18417088 TI - Does intraoperative radioguided surgery influence the complication rates and completeness of completion thyroidectomy? AB - BACKGROUND: To investigate whether radioguided surgery (RGS) has any beneficial effects on the complication rates and the completeness of completion thyroidectomy (CT) in a center experienced in endocrine surgery. METHODS: Thirty three patients scheduled for CT for thyroid carcinoma were randomly selected for 2 types of intervention. CT was performed by RGS following administration of 5 mCi technetium-99m in 15 patients (group 1) and with conventional surgical exploration without RGS in 18 patients (group 2). The duration of the CT, thyroid function tests, iodine-131 uptake at 24 hours at the third postoperative week, and complication rates were compared between groups 1 and 2. RESULTS: In groups 1 and 2, the duration of CT (63.3 +/- 7.5 vs 65 +/- 10.8 minutes, P = .7), postoperative serum thyrotropin-stimulating hormone (TSH) levels (43.9 +/- 17.5 mIU/L vs 36.8 +/- 8.6 mIU/L, P = .2), postoperative (131)I uptake at 24 hours (6.86 +/- 1.7 vs 7.0 +/- 1.3, P = .8), and complication rates (13.3% vs 5.6%, P = .6) showed no significant differences. CONCLUSION: RGS during CT offers no benefit over conventional surgical exploration with respect to operation time, complication rates, or completeness of surgery in a center experienced in endocrine surgery. However, it might be helpful for general surgeons who are less familiar with re-operative thyroid surgery. PMID- 18417089 TI - Equestrian injuries: incidence, injury patterns and risk factors for 10 years of major traumatic injuries. PMID- 18417090 TI - New immunomodulators in the treatment of Graves' ophthalmopathy. AB - Steroids have been used in the therapy of the moderate to severe forms of Graves' ophthalmopathy (GO) and other autoimmune diseases as they act only as general immunosuppressants. Previous work has shown that blocking the CD-20 receptor on B lymphocytes has significantly affected the clinical course of GO, by rapidly reducing inflammation and the degree of proptosis. We have studied nine patients with Graves' disease, of whom seven had active GO and two, with newly diagnosed hyperthyroidism, only mild lid signs. We also studied a group of 20 consecutive patients, treated with intravenous glucocorticoids (IVGC) according to a standard protocol. Patients treated with RTX (1000mg i.v. twice at two-week interval) and those treated with IVGC (500mg i.v. for 16 weeks) were studied monthly up to 12 months after the first drug infusion. By ophthalmological examination, GO was assessed by the clinical activity score (CAS) and by the NOSPECS score. Thyroid function and lymphocyte count were measured by standardized methods. RTX was well tolerated and only minor side-effects were reported in 30% of patients during the first infusion. All patients attained peripheral B-cell depletion with the first RTX infusion. All but one patients showed both CD20+ cells and CD19+ cells depletion, while one had persistent 3-5% CD19+ cells in the periphery, mostly CD19+5+. Thyroid function was not affected by RTX therapy. Titers of antithyroglobulin (TgAb), antithyroperoxidase and anti-TSH receptor antibodies (TRAb) did not change significantly (P=NS) and did not correlate to CD20+ depletion (P=NS). CAS values decreased significantly (P<0.0001). Proptosis decreased significantly after RTX in both patients with active GO (ANOVA; P<0.0001) and in those with Graves' hyperthyroidism and lid signs (ANOVA; P<0.003). The degree of inflammation (NOSPECS class 2) decreased significantly in response to RTX (ANOVA; P<0.001). In patients treated with IVGC, mean CAS value decreased significantly less than in those treated with RTX (P<0.05). Adverse effects were more frequent after IVGC (45% of patients). Seventy-five percent of patients responded to IVGC and 10% showed relapse of active GO six to eight weeks after withdrawal. The results of this study on RTX in GO suggest that the drug is effective in modifying the disease course and that the improvement of the clinical activity of GO after RTX was more significant than after IVGC. We did not observe relapse of active GO, even after B-cell return in peripheral blood. This might be related to the persistence of a significant degree of B-cell depletion after RTX observed in the peripheral blood as late as two years of follow-up. RTX therapy was also effective in improving proptosis and soft tissue inflammation. The mechanism by which RTX affects GO is unknown. It may act as a true immunosuppressor by switching off reactions inducing the active phase of TAO, perhaps by influencing the cytokine production in the orbit or by inducing depletion of antigen presenting B-cells. PMID- 18417091 TI - Molecular diagnosis of differentiated thyroid cancer. Towards the application in clinical practice. PMID- 18417092 TI - Preclinical pharmacology of the new GLP-1 receptor agonist AVE0010. PMID- 18417093 TI - [Hormonal targets of the autoimmune response]. PMID- 18417094 TI - Preclinical evidences suggest new treatment options for endocrine disorders: Pasireotide (SOM230) and Everolimus (RAD001). PMID- 18417095 TI - A change management framework for macroergonomic field research. AB - With the proliferation of macroergonomic field research, it is time to carefully examine how such research should be managed and implemented. We argue that the importance of attending to high-quality implementation of field research is equal to that of methodological rigor. One way to systematically manage the implementation process is to adopt a change management framework, wherein the research project is conceptualized as an instance of organization-level change. Consequently, principles for successful organization-level change from the literature on change management can be used to guide successful field research implementation. This paper briefly reviews that literature, deriving 30 principles of successful change management, covering topics such as political awareness, assembling the change team, generating buy-in, and management support. For each principle, corresponding suggestions for macroergonomic field research practice are presented. We urge other researchers to further develop and adopt frameworks that guide the implementation of field research. PMID- 18417096 TI - Defining stakeholder involvement in participatory design processes. AB - A participatory approach could be used to implement work place or organizational improvements. However, the question is which participants should be involved and how. In this paper the theoretical involvement in different steps of a linear stepwise approach is described and compared with the latest projects of 300 practitioners. From a theoretical point of view ergonomists and employees play an essential role in the improvement process and are involved in most stages of a change process. Designers play an important role in idea generation and prototyping. Top management and middle management are important in the first step to set goals that are consistent with the strategy of the enterprise. Middle management is also needed in the steps where improvements are selected. This theoretical prediction is affirmed. However, middle management appeared to be also involved in implementation. The role of ergonomists is in practice limited in later stages in implementation, which is not preferred by the ergonomists. PMID- 18417097 TI - Facilitating participatory steps for planning and implementing low-cost improvements in small workplaces. AB - In this paper, practical means of facilitating participatory steps taken in workplace improvement programs in small workplaces were reviewed. The reviewed programs included those organized by partners of our Asian inter-country network for small enterprises, construction sites, home workplaces and agricultural farms. Trainers who commonly acted as facilitators were found to play multiple roles in helping managers, workers and farmers take initiative and achieve immediate improvements. The participatory steps were more successfully facilitated when the trainers supported (a) building on local good practice, (b) focusing on a range of basic ergonomics principles, and (c) stepwise progress through feedback of achievements. The use of action-oriented toolkits comprising low-cost action checklists and group work guides was commonly helpful. The locally adjusted nature of the toolkits seemed essential. Trainers could thus help people build local initiative, plan and implement low-cost ideas through serial group work steps and confirm benefits in a stepwise manner. The review of the results suggested that a local network of trainers trained in the use of locally adjusted toolkits was vital for facilitating effective improvements in different small workplaces. PMID- 18417098 TI - Psychosocial and socio-demographic correlates of medication compliance among people with schizophrenia. AB - This study examined the medication compliance of people with schizophrenia in relation to their self-stigma, insight, attitude towards medication, and socio demographic status via a cross-sectional observational design. Eighty-six Chinese adults with schizophrenia were recruited from the psychiatric hospitals and community settings for this study. The findings suggested that stereotype agreement of self-stigmatization and attitude towards medication were moderately correlated with medication compliance. Poor insight and living alone were found to be significant predictors of medication compliance based on regression analysis. Insight was identified to be the strongest predictor on compliance which accounted for 68.35% of the total variance. Although self-stigma is only moderately linked with medication compliance, its effects on medication-induced stigma cannot be ignored. PMID- 18417099 TI - Daytime functioning in older patients suffering from chronic insomnia: treatment outcome in a randomized controlled trial comparing CBT with Zopiclone. AB - The paper presents data from a randomized controlled trial comparing treatment effects of cognitive behavioural therapy (CBT), hypnotic treatment (Zopiclone), and placebo in a sample of insomnia patients. Data from the same trial have already demonstrated that CBT was more efficient in improving sleep than Zopiclone. The novel outcomes that are reported here concern daytime functioning. Forty-six older patients (age >or= 55) qualifying for a diagnosis of primary insomnia were recruited to participate. Assessments were completed at baseline, post-treatment, and at a 6-months follow-up, and measures of worry, anxiety, depression, interpersonal relationships, subjective alertness, vigilance, and quality of life were used. The participants in both treatment conditions scored within the normal range on the outcome measures at baseline with the exception of reporting less alertness, relative to a group of good sleepers. One interaction effect indicated that subjective alertness improved more in the Zopiclone group than the CBT group from baseline to post-treatment, and another that CBT was more effective than Zopiclone in reducing trait anxiety from baseline to follow-up. It was concluded that the treatments yielded only minor effects on the measures of daytime functioning, and that none of them was clearly superior to the other. PMID- 18417100 TI - Accuracy of retrospective memory and covariation estimation in patients with obsessive-compulsive disorder. AB - Assessment methods relying on biased or inaccurate retrospective recall may distort knowledge about the nature of disorders and lead to faulty clinical inferences. Despite concerns about the accuracy of retrospective recall in general and in particular with obsessive-compulsive disorder (OCD) patients, the accuracy of retrospective recall for one's own symptoms assessed in vivo is unknown in this population. This study used a prospective ecological momentary assessment (EMA) methodology to create a criterion against which to assess recall accuracy in OCD patients. Although results indicated that patients' retrospective recall of OCD symptoms was fairly accurate, they consistently overestimated the magnitude of OCD symptom covariation with non-OCD facets (e.g., sleep duration, contemporaneous stress level, etc.). Findings suggest that even when recall of OCD symptoms is accurate, patients may be inaccurate in estimating symptom covariation. The findings have implications for the research, case conceptualization, and assessment of OCD, and may extend to other disorders. PMID- 18417101 TI - A cyclopentanediol analogue selectively suppresses the conserved innate immunity pathways, Drosophila IMD and TNF-alpha pathways. AB - Innate immunity comprises evolutionarily conserved self-defense mechanisms against microbial infections. In mammals, innate immunity interacts with adaptive immunity and has a key role in the regulated immune response. Therefore, innate immunity is a pharmaceutical target for the development of immune regulators. Using Drosophila ex vivo culture systems, we isolated a cyclopentanediol analogue from Aspergillus sp. as an immunosuppressive substance. This compound selectively suppressed activation of the IMD pathway in Drosophila in vivo and the target molecules of the compound lie between the Imd adaptor protein and dTAK1 kinase in the IMD pathway. In human cells, the compound suppressed TNF-alpha, but not IL 1beta, stimulation-induced activation of NF-kappaB, suggesting that its target molecules are upstream of TAK1 in mammalian innate immunity. PMID- 18417102 TI - Impaired fear memory, altered object memory and modified hippocampal synaptic plasticity in split-brain mice. AB - The hippocampus is critical for memory formation. However, the contributions of the hippocampal commissure (HC) and the corpus callosum (CC) are less clear. To elucidate the role of the forebrain commissures in learning and memory, we performed a behavioural and electrophysiological characterization of an inbred mouse strain that displays agenesis of the CC and congenitally reduced HC (BTBR T+ tf/J; 'BTBR'). Compared to a control strain, BTBR mice have severely impaired contextual fear memory, with normal object recognition memory. Interestingly, continuous environmental "enrichment" significantly increased object recognition in BTBR, but not in control C57BL/6 ('BL/6') mice. In area CA1 of hippocampal slices, BTBR displayed intact expression of long-term potentiation (LTP), paired pulse facilitation (PPF) and basal synaptic transmission, compared to BL/6 mice. However, BTBR hippocampal slices show an increased susceptibility to depotentiation (DPT), an activity-induced reversal of LTP. We conclude that the HC and CC are critical for some forms of hippocampal memory and for synaptic resistance to DPT. Agenesis of the CC and HC may unmask some latent ability to encode, store or retrieve certain forms of recognition memory. We suggest that the increased susceptibility to DPT in BTBR may underlie the memory phenotype reported here. PMID- 18417103 TI - Glucose transporter 5 is undetectable in outer hair cells and does not contribute to cochlear amplification. AB - Glucose transporter 5 (Glut5) is a high-affinity fructose transporter. It was proposed to be a motor protein or part of the motor complex required for cochlear amplification in outer hair cells (OHCs). Here we show that, in contrast to previous reports, Glut5 is undetectable, and possibly absent, in OHCs harvested from wildtype mice. Further, Glut5-deficient mice display normal OHC morphology and motor function (i.e., nonlinear capacitance and electromotility) and normal cochlear sensitivity and frequency selectivity. We conclude that Glut5 is not required for OHC motility or cochlear amplification. PMID- 18417104 TI - Involvement of serotonin 2A receptors in the analgesic effect of tramadol in mono arthritic rats. AB - The analgesic effects of tramadol are considered to be mediated by both the opioid system and the serotonergic system. This study investigated the involvement of a subtype of serotonin receptors, 5-hydroxytryptamine (5-HT)2A receptor, in the analgesic effect of tramadol. The intraperitoneal (i.p.) injection of tramadol reduced the paw withdrawal latency (PWL) to radiant heat testing in mono-arthritic rats. The antagonistic effect of i.p. ketanserin (a 5 HT2A receptor antagonist) on tramadol analgesia was observed. The expression of the 5-HT2A receptor mRNA in the nucleus of raphe magnus (NRM), ventrolateral periaqueductal gray (vlPAG) and spinal dorsal horn of mono-arthritic rats after a ten-day treatment with tramadol was measured with in situ hybridization. Either single injections or 10 days of tramadol treatment dose-dependently elevated PWL of arthritic rats while ketanserin could partially antagonize the tramadol analgesic effect. Expression of the 5-HT2A receptor mRNA in NRM, ipsilateral vlPAG, and the ipsilateral spinal dorsal horn of arthritic rats was significantly increased after tramadol treatment. These results suggest that 5-HT2A receptors are involved in the analgesic effect of tramadol. This study provides evidence for involvement of 5-HT2A receptors in the tramadol analgesia of inflammatory pain. The increase in this receptor mRNA in the chronic study may contribute to the sustaining effect of tramadol long-term treatments in clinical practice. PMID- 18417105 TI - Cocaine withdrawal-induced trafficking of delta-opioid receptors in rat nucleus accumbens. AB - Interactions between the opioidergic and dopaminergic systems in the nucleus accumbens (NAcb) play a critical role in mediating cocaine withdrawal-induced effects on cell signaling and behavior. In support of this, increased activation of striatal dopamine-D1 receptors (D1R) results in desensitization of delta opioid receptor (DOR) signaling through adenylyl cyclase during early cocaine withdrawal. A potential cellular substrate underlying receptor desensitization is receptor internalization. The present study examined the effect of cocaine withdrawal on subcellular localization of DOR in dendrites of the NAcb core (NAcbC) and shell (NAcbS) using immunoelectron microscopy. Female and male rats received binge-pattern cocaine or saline for 14 days and subsequently underwent 48 h withdrawal. Animals were transcardially perfused and tissue sections were processed for immunogold-silver localization of DOR. Semi-quantitative analysis revealed that cocaine withdrawal caused an increase in the percentage of DOR localized intracellularly in the NAcbS of male and female rats and the NAcbC of male rats compared to saline controls. In contrast, in the NAcbC of female rats, there was an increase in DOR associated with the plasma membrane following cocaine withdrawal. To determine whether modulation of D1R could directly impact DOR containing neurons, the hypothesis that DOR and D1R co-exist in common neurons of the NAcb was examined in naive rats. Semi-quantitative analysis revealed a subset of profiles containing both DOR and D1R immunoreactivities. The present findings demonstrate a redistribution of DOR in the NAcb following cocaine withdrawal and provide anatomical evidence supporting D1R regulation of DOR function in a subset of NAcb neurons. PMID- 18417107 TI - Automatic lung nodule matching on sequential CT images. AB - We propose an automatic segmentation and registration method that provides more efficient and robust matching of lung nodules in sequential chest computed tomography (CT) images. Our method consists of four steps. First, the lungs are extracted from chest CT images by the automatic segmentation method. Second, gross translational mismatch is corrected by optimal cube registration. This initial alignment does not require extracting any anatomical landmarks. Third, the initial alignment is step-by-step refined by hierarchical surface registration. To evaluate the distance measures between lung boundary points, a three-dimensional distance map is generated by narrow-band distance propagation, which drives fast and robust convergence to the optimal value. Finally, correspondences of manually detected nodules are established from the pairs with the smallest Euclidean distances. Experimental results show that our segmentation method accurately extracts lung boundaries and the registration method effectively finds the nodule correspondences. PMID- 18417106 TI - The role of category learning in the acquisition and retention of perceptual expertise: a behavioral and neurophysiological study. AB - This study examined the neural mechanisms underlying perceptual categorization and expertise. Participants were either exposed to or learned to classify three categories of cars (sedans, SUVs, antiques) at either the basic or subordinate level. Event-Related Potentials (ERPs) as well as accuracy and reaction time were recorded before, immediately after, and 1-week after training. Behavioral results showed that only subordinate-level training led to better discrimination of trained cars, and this ability was retained a week after training. ERPs showed an equivalent increase in the N170 across all three training conditions whereas the N250 was only enhanced in response to subordinate-level training. The behavioral and electrophysiological results distinguish category learning at the subordinate level from category learning occurring at the basic level or from simple exposure. Together with data from previous investigations, the current results suggest that subordinate-level training, but not basic-level or exposure training, leads to expert-like improvements in categorization accuracy. These improvements are mirrored by changes in the N250 rather than the N170 component, and these effects persist at least a week after training, so are conceivably related to long-term learning processes supporting perceptual expertise. PMID- 18417108 TI - Anteroventrally localized activity in the optic vesicle plays a crucial role in the optic development. AB - The vertebrate eye develops from the optic vesicle (OV), a laterally protrusive structure of the forebrain, by a coordinated interaction with surrounding tissues. The OV then invaginates to form an optic cup, and the lens placode develops to the lens vesicle at the same time. These aspects in the early stage characterize vertebrate eye formation and are controlled by appropriate dorsal ventral coordination. In the present study, we performed surgical manipulation in the chick OV to remove either the dorsal or ventral half and examined the development of the remaining OV. The results show that the dorsal and ventral halves of the OV have a clearly different developmental pattern. When the dorsal half was removed, the remaining ventral OV developed into an entire eye, while the dorsal OV developed to a pigmented vesicle consisting of retinal pigmented epithelium alone. These results indicate that the ventral part of the OV retains the potency to develop the entire eye structure and plays an essential role in proper eye development. In subsequent manipulations of early chick embryos, it was found that only the anterior ventral quadrant of the OV has the potential to develop the entire eye and that no other part of the OV has a similar activity. Fgf8 expression was localized in this portion and no Fgf8 expression was observed within the OV when the ventral OV was removed. These results suggest that the anterior ventral portion of the OV plays a crucial role in the proper development of the eye, possibly generating the dorsal-ventral gradients of signal proteins within the eye primordium. PMID- 18417109 TI - Sequential actions of Pax3 and Pax7 drive xanthophore development in zebrafish neural crest. AB - The Pax3/7 gene family has a fundamental and conserved role during neural crest formation. In people, PAX3 mutation causes Waardenburg syndrome, and murine Pax3 is essential for pigment formation. However, it is unclear exactly how Pax3 functions within the neural crest. Here we show that pax3 is expressed before other pax3/7 members, including duplicated pax3b, pax7 and pax7b genes, early in zebrafish neural crest development. Knockdown of Pax3 protein by antisense morpholino oligonucleotides results in defective fate specification of xanthophores, with complete ablation in the trunk. Other pigment lineages are specified and differentiate. As a consequence of xanthophore loss, expression of pax7, a marker of the xanthophore lineage, is reduced in neural crest. Morpholino knockdown of Pax7 protein shows that Pax7 itself is dispensable for xanthophore fate specification, although yellow pigmentation is reduced. Loss of xanthophores after reduction of Pax3 correlates with a delay in melanoblast differentiation followed by significant increase in melanophores, suggestive of a Pax3-driven fate switch within a chromatophore precursor or stem cell. Analysis of other neural crest derivatives reveals that, in the absence of Pax3, the enteric nervous system is ablated from its inception. Therefore, Pax3 in zebrafish is required for specification of two specific lineages of neural crest, xanthophores and enteric neurons. PMID- 18417111 TI - Absorption of polycyclic aromatic hydrocarbons by Pinus bark: analytical method and use for environmental pollution monitoring in the Palermo area (Sicily, Italy). AB - In the light of using biomaterial as pollutants sorbents for the environmental biomonitoring, we report here the results of the absorption of polycyclic aromatic hydrocarbons (PAHs) by pine bark. Quantitative analysis of 19 different polycyclic aromatic compounds was performed, after Soxhlet extraction in dichloromethane, by means GC-MS technique. Pinus was chosen due to its wide distribution in the Mediterranean area. The passive absorption of PAH by pine bark in the metropolitan areas allowed us to evaluate the air quality of Palermo. The obtained results showed a very high range of PAHs concentrations, from 33 microg/kg (bark dry weight), as SigmaPAHs, at the control site to 1015 microg/kg along a road with high traffic flow. PMID- 18417110 TI - Vsx2/Chx10 ensures the correct timing and magnitude of Hedgehog signaling in the mouse retina. AB - Vertebrate retinal progenitor cells (RPCs) undergo a robust proliferative expansion to produce enough cells for the retina to form appropriately. Vsx2 (formerly Chx10), a homeodomain protein expressed in RPCs, is required for sufficient proliferation to occur. Sonic Hedgehog protein (SHH), secreted by retinal ganglion cells (RGCs), activates Hedgehog (Hh) signaling in RPCs and is also required for sufficient proliferation to occur. Therefore, we sought to determine if reduced Hh signaling is a contributing factor to the proliferation changes that occur in the absence of Vsx2. To do this, we examined Shh expression and Hh signaling activity in the homozygous ocular retardation J (orJ) mouse, which harbors a recessive null allele in the Vsx2 gene. We found that Shh expression and Hh signaling activity are delayed during early retinal development in orJ mice and this correlates with a delay in the onset of RGC differentiation. At birth, reduced expression of genes regulated by Hh signaling was observed despite the production of SHH ligand. orJ RPCs respond to pre-processed recombinant SHH ligand (SHH-N) in explant culture as evidenced by increased proliferation and expression of Hh target genes. Interestingly, proliferation in the orJ retina is further inhibited by cyclopamine, an antagonist of Hh signaling. Our results suggest that reduced Hh signaling contributes to the reduced level of RPC proliferation in the orJ retina, thereby revealing a role for Vsx2 in mediating mitogen signaling. PMID- 18417112 TI - Mechanisms of renal anionic drug transport. AB - By utilizing filtration, active secretion and reabsorption processes, the kidney can conserve essential nutrients, and eliminate drugs and potentially toxic compounds. Active uptake of organic anions and cations across the basolateral membrane, and their extrusion into the urine across the brush border membrane mainly takes place in the renal proximal tubule cells, and is facilitated via a range of substrate-specific tubular transporters. Many drugs and their phase II conjugates are anionic compounds, and therefore renal organic anion transporters are important determinants of their distribution and elimination. Competition for renal excretory transporters may cause drugs to accumulate in the body leading to toxicity, which is a potential hazard of concomitant drug administration. Here, we present a brief update on the most prominent human proximal tubule organic anion transporters, which either belong to the ATP-binding cassette (ABC) or the solute carrier transporter (SLC) families. We focus on the participation of the individual transporters in renal anionic drug elimination, in an attempt to understand their overall biological and pharmacological significance, hoping to inspire further studies in the renal transporters field. PMID- 18417113 TI - The (pro)renin receptor: a new addition to the renin-angiotensin system? AB - The renin-angiotensin system is still incompletely understood. In particular, the function of prorenin, the inactive precursor of renin, is unknown. Yet, prorenin levels are >10-fold higher than renin levels, and prorenin increases even further in subjects with diabetes mellitus displaying microvascular complications. The recent discovery of a (pro)renin binding receptor may shed light on the role of prorenin. This review discusses the possibility that prorenin binding to this receptor results in prorenin activation, thereby allowing angiotensin generation, and that prorenin simultaneously acts as an agonist of this receptor, inducing angiotensin-independent effects. Transgenic animals overexpressing the receptor, as well as a receptor antagonist are now available, and future studies should reveal to what degree this concept is applicable to humans as well. PMID- 18417114 TI - Pharmacology of airway smooth muscle proliferation. AB - Airway smooth muscle thickening is a pathological feature that contributes significantly to airflow limitation and airway hyperresponsiveness in asthma. Ongoing research efforts aimed at identifying the mechanisms responsible for the increased airway smooth muscle mass have indicated that hyperplasia of airway smooth muscle, due in part to airway myocyte proliferation, is likely a major factor. Airway smooth muscle proliferation has been studied extensively in culture and in animal models of asthma, and these studies have revealed that a variety of receptors and mediators contributes to this response. This review aims to provide an overview of the receptors and mediators that control airway smooth muscle cell proliferation, with emphasis on the intracellular signalling mechanisms involved. PMID- 18417115 TI - Neuro-immune interactions in inflammatory bowel disease and irritable bowel syndrome: future therapeutic targets. AB - The gastro-intestinal tract is well known for its largest neural network outside the central nervous system and for the most extensive immune system in the body. Research in neurogastroenterology implicates the involvement of both enteric nervous system and immune system in symptoms of inflammatory bowel disease and irritable bowel syndrome. Since both disorders are associated with increased immune cell numbers, nerve growth and activation of both immune cells and nerves, we focus in this review on the involvement of immune cell-nerve interactions in inflammatory bowel disease and irritable bowel syndrome. Firstly, the possible effects of enteric nerves, especially of the nonadrenergic and noncholinergic nerves, on the intestinal immune system and their possible role in the pathogenesis of chronic intestinal inflammatory diseases are described. Secondly, the possible effects of immunological factors, from the innate (chemokines and Toll-like receptors) as well as the adaptive (cytokines and immunoglobulins) immune system, on gastro-intestinal nerves and its potential role in the development of inflammatory bowel disease and irritable bowel syndrome are reviewed. Investigations of receptor-mediated and intracellular signal pathways in neuro-immune interactions might help to develop more effective therapeutic approaches for chronic inflammatory intestinal diseases. PMID- 18417116 TI - Health effects of quercetin: from antioxidant to nutraceutical. AB - Quercetin, a member of the flavonoids family, is one of the most prominent dietary antioxidants. It is ubiquitously present in foods including vegetables, fruit, tea and wine as well as countless food supplements and is claimed to exert beneficial health effects. This includes protection against various diseases such as osteoporosis, certain forms of cancer, pulmonary and cardiovascular diseases but also against aging. Especially the ability of quercetin to scavenge highly reactive species such as peroxynitrite and the hydroxyl radical is suggested to be involved in these possible beneficial health effects. Consequently, numerous studies have been performed to gather scientific evidence for these beneficial health claims as well as data regarding the exact mechanism of action and possible toxicological aspects of this flavonoid. The purpose of this review is to evaluate these studies in order to elucidate the possible health-beneficial effects of the antioxidant quercetin. Firstly, the definitions as well as the most important aspects regarding free radicals, antioxidants and oxidative stress will be discussed as background information. Subsequently, the mechanism by which quercetin may operate as an antioxidant (tested in vitro) as well as the potential use of this antioxidant as a nutraceutical (tested both ex vivo and in vivo) will be discussed. PMID- 18417117 TI - Angiotensin-(1-7): pharmacological properties and pharmacotherapeutic perspectives. AB - Therapeutic modulation of the renin-angiotensin system is not complete without taking into consideration the beneficial effects of angiotensin-(1-7) in cardiovascular pathology. Various pharmacological pathways are already exploited to involve this heptapeptide in therapy as both inhibitors of angiotensin converting enzyme and angiotensin II type 1 receptor blockers increase its levels. These drugs and administered angiotensin-(1-7) elicit various common effects, and some effects of the drugs are partially mediated by angiotensin-(1 7). The pharmacodynamic profile of angiotensin-(1-7) is rather complex, and in vitro and in vivo studies demonstrated a wide palette of effects for angiotensin (1-7), some of them potentially beneficial for cardiovascular disease. Using various animal models to study cardiovascular physiology and disease it was shown that angiotensin-(1-7) has antihypertensive, antihypertrophic, antifibrotic and antithrombotic properties, all properties that may prove beneficial in a clinical setting. We also observed a novel action of angiotensin-(1-7), namely its capacity to stimulate the proliferation of endothelial progenitor cells. Access of angiotensin-(1-7) to the clinic, however, is restricted due to its unfavorable pharmacokinetic properties. In order to benefit of the therapeutic potential of angiotensin-(1-7) it is crucial to increase its half-life, either by using more stable analogues, which are now under development, or specific delivery methods. We here review the pharmacological characteristics and therapeutic potential of angiotensin-(1-7), implementing the experimental strategies taken to exploit the pharmacological mechanism of this heptapeptide in a clinical setting, and present our contribution to this field of research. PMID- 18417118 TI - Implications of hepatic cytochrome P450-related biotransformation processes in veterinary sciences. AB - Cytochrome P450 enzymes (CYP450) represent a superfamily of monooxigenases that play a pivotal role in drug metabolism. In contrast to the extensive database available for human and rodent CYP450 enzyme activities, the data related to animal species that are regular patients in veterinary medicine, are far from being complete. The major obstacles are the significant inter-species and intra species differences. With the aim to provide an overview of the current knowledge, key data for important species, such as dogs and cats, horses, pigs and ruminants, are presented, and compared with findings from humans. Analysis of these data shows, that currently no links can be established between certain physiological traits, such as herbivorous and carnivorous species, monogastric animal and ruminants, nor within a given species, as for example cattle. This implies that for all new pharmaceutical entities individual assays are needed for every animal species or even every individual breed. It can be anticipated, however, that investigations into the upstream transcriptional regulation of CYP450 enzymes will provide more insight into the observed expression levels, thus allowing to modulate kinetic parameters of old and new drugs, as the same transcription factors control also the expression of prominent drug transporters. PMID- 18417119 TI - Implications of ABC transporters on the disposition of typical veterinary medicinal products. AB - The ATP-Binding Cassette (ABC) transporters ABCB1, ABCC2 and ABCG2 are efflux transporters that facilitate the excretion of drugs, contribute to the function of biological barriers and maintain low cytoplasmic substrate concentrations in cells. ABC transporters modulate drug absorption, distribution and elimination according to the level of expression in the intestine, liver, kidney, and at biological barriers such as the blood-brain barrier. Moreover individual transporters are known to convey multi-drug resistance to tumour cells. While these diverse functions have been described in laboratory animal studies and in humans, the available information is very limited in animal species that are typical veterinary patients. This brief review summarizes the available data on organ distribution and expression levels in animals, genetic defects in dogs resulting in a non-functional P-gp expression, and describes examples of kinetic investigations directed to assess the clinical relevance of species differences in ABC-transporter expression. PMID- 18417120 TI - Neuropharmacology of glucocorticoids: focus on emotion, cognition and cocaine. AB - Hormone pharmacology has been quite interesting in The Netherlands the past century and this contribution is dedicated to the glucocorticoid hormones underlying adaptation to stress. The story starts in 1936 with Tadeus Reichstein and Ernst Laqueur who discovered corticosterone at the time Hans Selye formulated the stress concept. Today highly sophisticated technologies help to unravel the action mechanism of the glucocorticoids from gene to behaviour. In today's concept glucocorticoids coordinate in concert with other stress mediators the initial stress reactions with the management of later adaptations. Glucocorticoids modulate early life programming of stress reactivity and are a significant factor in brain plasticity underlying adaptation, the aging process and vulnerability to disease. Here we focus on the role of glucocorticoids in emotions, cognitive performance and behavioural sensitisation to cocaine. PMID- 18417121 TI - The Wnt/frizzled pathway in cardiovascular development and disease: friend or foe? AB - Proteins from the Wnt family have been implicated in cell-cell communication in a wide variety of developmental and physiological processes. Wnt signaling is required for different aspects of cardiac and vascular development, including myocardial specification, cardiac morphogenesis and cardiac valve formation as well as endothelial and vascular smooth muscle cell proliferation. Defective Wnt signaling can result in different cardiac and vascular abnormalities. In the adult heart and blood vessels, Wnt signaling activity is quite low under normal conditions. However, this pathway is reactivated during the pathological cardiac remodeling induced by pressure overload, in injured arteries and after myocardial infarction. Genetically modified animal models have shown that inhibition of Wnt signaling results in increased angiogenesis, better infarct healing and an attenuated hypertrophic response of the heart. This suggests that pharmacological inhibition of Wnt signaling could provide a novel therapeutic strategy to prevent excessive cardiac and vascular remodeling. PMID- 18417122 TI - Tissue transglutaminase: a novel pharmacological target in preventing toxic protein aggregation in neurodegenerative diseases. AB - Alzheimer's disease, Parkinson's disease and Huntington's disease are neurodegenerative diseases, characterized by the accumulation and deposition of neurotoxic protein aggregates. The capacity of specific proteins to self-interact and form neurotoxic aggregates seems to be a common underlying mechanism leading to pathology in these neurodegenerative diseases. This process might be initiated and/or accelerated by proteins that interact with these aggregating proteins. The transglutaminase (TG) family of proteins are calcium-dependent enzymes that catalyze the formation of covalent epsilon-(gamma-glutamyl)lysine isopeptide bonds, which can result in both intra- and intermolecular cross-links. Intramolecular cross-links might modify self-interacting proteins, and make them more prone to aggregate. In addition, intermolecular cross-links could link self aggregating proteins and thereby initiate and/or stimulate the aggregation process. So far, increased levels and activity of tissue transglutaminase (tTG), the best characterized member of the TG family, have been observed in many neurodegenerative diseases, and the self-interacting proteins, characteristic of Alzheimer's disease, Parkinson's disease and Huntington's disease, are known substrates of tTG. Here, we focus on the role of tTG in the initiation of the aggregation process of self-interacting proteins in these diseases, and promote the notion that tTG might be an attractive novel target for treatment of neurodegenerative diseases. PMID- 18417123 TI - Human in vivo research on the vascular effects of adenosine. AB - In situations of impending tissue danger, such as during ischaemia, the concentration of the endogenous purine nucleoside adenosine rapidly increases. Subsequent stimulation of G-protein coupled adenosine receptors induces several cardiovascular effects, such as vasodilation, inhibition of inflammation, modulation of sympathetic nervous system activity, and increasing myocardial tolerance against ischaemia-reperfusion, which are all aimed at protecting the affected tissue. Although animal models have consistently shown profound cardiovascular protection by adenosine, up to now translation of this knowledge into clinical practice is limited. This current review is focused on human in vivo studies on the cardiovascular effects of adenosine. Several techniques, such as microdialysis, venous occlusion plethysmography, and (99m)Tc-annexin A5 scintigraphy can be used to study these effects of adenosine in healthy volunteers in vivo. By use of these techniques, recent studies have shown that the cardiovascular effects of adenosine can be modulated by genetic factors (e.g. a single nucleotide polymorphism in the gene encoding for adenosine monophosphate deaminase), by metabolic factors (e.g. by the plasma homocysteine concentration), and by drugs, such as caffeine, dipyridamole, and methotrexate. Given the cardiovascular protective properties of adenosine, these factors could well modulate the risk or extent of cardiovascular disease in patients and knowledge of these factors could be of benefit in daily clinical practice to optimize treatment in patients with cardiovascular disease. PMID- 18417124 TI - Osteoprotegerin/RANK/RANKL axis in cardiac remodeling due to immuno-inflammatory myocardial disease. AB - OBJECTIVE: Osteoprotegerin (OPG) and the receptor activator of nuclear factor kappaB ligand (RANKL) are inflammatory cytokines traditionally linked to the regulation of bone remodeling. We hypothesize that the OPG/RANK/RANKL axis may be involved in extracellular matrix remodeling in immuno-inflammatory heart diseases, and explore the probable underlying mechanisms by using anti-IL-17 in the model of experimental autoimmune myocarditis. METHODS: EAM was induced in Lewis rats by injection of porcine cardiac myosin. All the rats were randomly distributed into day 0, day 7, day 14 and day 28 groups, which means rats in certain group were cervical dislocated on day 0, day 7, day 14 and day 28 respectively. HE staining and Masson's staining were used for measurement of cardiac hypertrophy and interstitial fibrosis. Hydroxyproline content and collagen cross-linking were determined in heart section. Anti-IL-17 or control antibody was injected i.p. 2 h before and 3 days and 7 days after the first myosin immunization in EAM model, that is, group anti-IL-17 or group control antibody. IL-17 and OPG/RANK/RANKL axis expressions were detected by realtime RT PCR in all the six groups. In the in vitro studies, cardiac fibroblasts were cultured and treated with IL-17 or vehicle for 48 h. Total RNA was isolated from harvested cells and realtime RT-PCR was performed to detect the RANK, RANKL, OPG and MMP-2, MMP-9, TIMP-1 and TIMP-2 expressions, then matrix metalloproteinase activity was assayed. RESULTS: Our in vivo results revealed that expression of IL 17 and the OPG/RANK/RANKL axis increased significantly from day 0 to day 28, with IL-17 and OPG increased relatively steeply. In the in vitro study, we detected OPG, RANK and RANKL mRNA expressions in the cultured fibroblasts with or without IL-17 stimulation. We found that IL-17 increased the OPG/RANK/RANKL axis activity (P<0.05). Although IL-17 induced a significant increase in MMP-2 and MMP-9 gene expressions in cardiac fibroblasts, there was no change in TIMP-2 and TIMP-1 expressions. CONCLUSIONS: Our results suggest that the OPG/RANK/RANKL axis may be involved in cardiac remodeling in immuno-inflammatory myocardial diseases and progression of chronic HF and thus may represent targets for intervention in this disorder. PMID- 18417125 TI - Enhanced function in the good forelimb of hemi-parkinson rats: compensatory adaptation for contralateral postural instability? AB - In this paper we present two new assays of rat motor behavior which can be used to assess function linked to postural stability in each forelimb independently. Postural instability is a major deficit in Parkinson's disease that is resistant to levodopa therapy and contributes to the risk of falling. We applied both tests, one forelimb at a time, to normal rats as well as rats extensively depleted of dopamine by unilateral infusion of 6-hydroxydopamine (6-OHDA, given in the medial forebrain bundle) to produce a hemi-parkinsonian syndrome. The 6 OHDA rats showed severe postural instability in the impaired forelimb, but unexpectedly showed enhanced function in the non-impaired forelimb. The data suggest that the intact hemisphere may undergo rapid reorganization subsequent to unilateral dopamine depletion, which allows for compensatory function of the "intact" limb. Measurements of amphetamine-induced striatal c-fos expression, as well as behavior results gathered when animals were under the influence of apomorphine or haloperidol, indicate that this potential reorganization may require non-dopaminergic neural plasticity. The relevance of these findings for unilateral rat models of neurological disease is discussed. PMID- 18417126 TI - The false and the true bifid condyles. AB - The bifid mandibular condyle has been described as a condition of unknown aetiology and uncertain pathogenesis. Many see it as the product of accidental trauma or forceps delivery, with the two heads occurring one behind the other in the sagittal plane. In bioanthropological literature, "bifid condyle" often describes pitting in the sagittal plane, dividing the condyle mediolaterally. We examined 38 male and 16 female pre-European-contact Pacific islanders' adult mandibles, and 24 male and 29 female modern Indian mandibles, recording frequency, prominence and position of any condylar groove in both coronal and sagittal planes. We report the tenth known case of a bilaterally-bifid condyle. A groove was found almost twice as likely to occur on the left condyle of the Indians than of the Pacific Islanders, but equally likely to occur on the right side of both groups. That same finding applied to males and females. In order to avoid terminological ambiguity, we suggest that the term "bifid condyle" should be reserved for describing multiple condyles in the sagittal plane only - the true bifid condyle. An hypothesis is offered for the occurrence of the groove in the sagittal plane. PMID- 18417127 TI - Maternal influences on the sexual behavior and reproductive success of the female rat. AB - In many species, including humans, there is evidence for parental effects on within-sex variations in reproductive behavior. In the present studies we found that variations in postnatal maternal care were associated with individual differences in female sexual behavior in the rat. Females born to and reared by dams that showed enhanced pup licking/grooming (i.e., High LG mothers) over the first week postpartum showed significantly reduced sexual receptivity and alterations in the pacing of male mounting (i.e., longer inter-intromission intervals) observed in a paced mating test. There were minimal effects on the sexual behavior of the male offspring. The female offspring of High LG mothers showed a reduced lordosis rating, a decreased mount:intromission ratio, received fewer ejaculations and were less likely to achieve pregnancy following mating in the paced mating context. The data suggest maternal influences on the sexual development of the female rat that are functionally relevant for reproductive success. Together with previous studies these findings imply that maternal care can 'program' reproductive strategies in the female rat. PMID- 18417128 TI - Patterns of testosterone responsiveness and immunity in relation to competitive behavior in chicks. PMID- 18417129 TI - Facilitating in-hospital transport of trauma patients: design of a trauma life support trolley. PMID- 18417130 TI - External adjuncts to enhance fracture healing: what is the role of ultrasound? AB - Current methods of fracture care use various adjuncts aimed at decreasing time to fracture union and improving fracture union rates. Among the most commonly used modalities, low-intensity pulsed ultrasound is emerging as a safe, cost-effective and reliable treatment for both fresh fractures and fracture nonunions. Both in vivo and in vitro basic science studies have helped to elucidate potential mechanisms of ultrasound action and a number of prospective, randomised, double blind, placebo-controlled trials exist demonstrating the clinical efficacy of low intensity pulsed ultrasound. This article will review the evidence for the use of low-intensity pulsed ultrasound in fracture care. PMID- 18417131 TI - Expandable nailing system for tibial shaft fractures. AB - OBJECTIVE: To evaluate and present our current clinical experience in the treatment of closed and open tibial shaft fractures using the expandable intramedullary nailing system. DESIGN: Prospective study. SETTING: One level-1 trauma centre. PARTICIPANTS: Twenty-six patients with acute tibial shaft fractures with at least 10 cm of intact cortex on both sides of the fracture. INTERVENTION: Internal fixation using the Fixion expandable intramedullary nail (Disc-O-Tech Medical Technologies Ltd., Herzliya, Israel). MAIN OUTCOME MEASUREMENTS: Operative and fluoroscopy time, healing time and perioperative complications were recorded. RESULTS: Twenty-six tibial fractures were treated (OTA classification: 3 type A1, 8 A2, 8 A3, 3 B1, 3 B2, and 1 B3), six of which were open. All fractures healed by week 18.5 with an mean of 12.8+/-3.8 weeks. The operating time ranged from 20 to 50 min with an mean of 40+/-12.17 min. The fluoroscopy time ranged from 6 to 22s with a mean of 10+/-5s. In one case the nail failed to expand, as detected by X-ray control, and had to be exchanged intraoperatively. Two patients reported anterior knee pain during the follow up, but did not wish any further treatment. In a low demanding patient rotational malalignment was noted and no further action needed. CONCLUSION: According to the results of this study, the use of the Fixion nailing system, where indicated, is associated with minimal complications and very good functional outcomes in fractures OTA types A and B. The advantages of the expandable nail include the decrease in the operating and fluoroscopy time and the simplicity of its application. PMID- 18417132 TI - Outcomes and costs of penetrating trauma injury in England and Wales. AB - BACKGROUND: Penetrating trauma injury is generally associated with higher short term mortality than blunt trauma, and results in substantial societal costs given the young age of those typically injured. Little information exists on the patient and treatment characteristics for penetrating trauma in England and Wales, and the acute outcomes and costs of care have not been documented and analysed in detail. METHODS: Using the Trauma Audit Research Network (TARN) database, we examined patient records for persons aged 18+ years hospitalised for penetrating trauma injury between January 2000 and December 2005. Patients were stratified by injury severity score (ISS). RESULTS: 1365 patients were identified; 16% with ISS 1-8, 50% ISS 9-15, 15% ISS 16-24, 16% ISS 25-34, and 4% with ISS 35-75. The median age was 30 years and 91% of patients were men. Over 90% of the injuries occurred in alleged assaults. Stabbings were the most common cause of injury (73%), followed by shootings (19%). Forty-seven percent were admitted to critical care for a median length of stay of 2 days; median total hospital length of stay was 7 days. Sixty-nine percent of patients underwent at least one surgical procedure. Eight percent of the patients died before discharge, with a mean time to death of 1.6 days (S.D. 4.0). Mortality ranged from 0% among patients with ISS 1-8 to 55% in patients with ISS>34. The mean hospital cost per patient was pound 7983, ranging from pound 6035 in patients with ISS 9-15 to pound 16,438 among patients with ISS>34. Costs varied significantly by ISS, hospital mortality, cause and body region of injury. CONCLUSION: The acute treatment costs of penetrating trauma injury in England and Wales vary by patient, injury and treatment characteristics. Measures designed to reduce the incidence and severity of penetrating trauma may result in significant hospital cost savings. PMID- 18417133 TI - Subperiosteal elevation of the ulnar nerve-Anatomical considerations and preliminary results. AB - Mobilisation of the ulnar nerve often leads to perineural scarring. About 30 years ago, B.F. Korosec, M.D., developed a novel surgical approach to the distal humerus combined with osteotomy of the olecranon, which avoids visualisation or direct manipulation of the ulnar nerve. The main advantage of his technique lies in subperiosteal elevation of the ulnar nerve together with all structures of the cubital tunnel. In the present study, anatomical dissections of distal humerus structures were performed in a cadaver to define individual steps of the technique introduced by Dr. Korosec. Ten phases of the procedure were identified. Ten patients (4 males and 6 females) with type 13-C and 13-B fractures of the distal humerus (AO/ASIF) were operated on. The patients showed no clinical signs of ulnar nerve injury before surgery. During the operation, the ulnar nerve was electrically stimulated at the axilla and compound muscle action potentials were continuously recorded from hypothenar muscles. Latency and amplitude of the potentials were monitored. Analysis of these data using the unpaired t-test for median values revealed no statistically significant differences between individual stages of the operation (p<0.00625). An amplitude decrease accompanied by a latency increase by more than 10% over the baseline value was found to indicate impending nerve damage. The novel approach to the distal humerus presented in this article is a safe procedure. PMID- 18417134 TI - Humeral nailing revisited. AB - Unreamed interlocked humeral nailing for stabilisation of acute humeral fractures was introduced a decade ago. Antegrade and retrograde nail insertion are equally popular. The role of nailing as opposed to plating of humeral fractures is the subject of continuous debate. Between 1997 and 2005, 99 acute fractures of the humeral shaft were treated operatively with the unreamed humeral nail (UHN, Synthes) in our Level I Trauma Centre. The mean age of the patients was 63 years. Only eight patients (8.1%) were polytraumatised, nine patients had an open fracture (9.1%), five had a primary radial nerve palsy (5.1%). There were 54 antegrade and 45 retrograde nailings. The procedures were performed by 19 different surgeons, who carefully followed a detailed operation protocol. There were 6 adverse events: 3 secondary radial nerve palsies (3%), 2 fissures at the insertion point (2%) and one false placement of a locking screw (1%). Three patients developed pseudarthrosis (3%). Eight further operation were necessary (8.1%): 3 exploration of the radial nerve, 3 for treatment of pseudarthrosis, one replacement of a locking screw and one wound revision for superficial wound infection. Ninety patients (92 fractures) were evaluated after bone healing. Shoulder function was assessed using the Constant Score, elbow function with the Mayo Elbow Score. 91.3% and 5.4% of patients had an excellent or good shoulder function, 81.5% and 14.1% had an excellent or good elbow function. All patients with a functional deficit of the shoulder joint had antegrade, all patients with a deficit at the elbow joint retrograde nailing. Motor function recovered in all radial nerve palsies. 93.5% of patients had an excellent or good functional end result. Unreamed humeral nailing is a valid therapeutic option for stabilisation of acute humeral shaft fractures. Antegrade and retrograde nailing are associated with specific but different complications. By strictly adhering to the operation technique, the number and severity of complications can be reduced. When good fracture alignment and stability are obtained, uneventful bone healing with good functional outcome is the rule. PMID- 18417135 TI - Double ureter at repair of obstetric fistula. PMID- 18417136 TI - Announcing the winner of the John J. Sciarra IJGO Prize Paper Award for 2007. PMID- 18417137 TI - Optimal waist circumference cutoffs for abdominal obesity in Chinese. AB - OBJECTIVE: To determine the appropriate cutoffs for visceral fat area (VFA) measured by magnetic resonance imaging linking to risk of the metabolic syndrome (MetS) and the corresponding waist circumference in Chinese. METHODS AND RESULTS: Totally 1,140 individuals (men 525, women 615) aged from 35 to 75 years were included. The components of the MetS were defined by International Diabetes Federation (IDF) and Chinese Diabetes Society (CDS) definition, respectively. Receive operating characteristic curve analyses were used to determine the appropriate cutoffs of VFA and corresponding waist circumference in the prediction of the MetS. The optimal VFA cutoff was near 80 cm(2) in identifying the MetS with two or more components but not including overweight/obesity by either of the two definitions in all subjects. There was no difference in men by ages while women aged < 50 years tended to have lower VFA cutoff than those aged > or = 50 years by the two definitions. The appropriate waist circumference cutoffs were 90 cm in men and 85 cm in women for the MetS. CONCLUSION: The optimal cutoff of waist circumference for abdominal obesity is 90 cm for men and 85 cm for women in Chinese. PMID- 18417138 TI - Synthesis of natural arm swing motion in human bipedal walking. AB - It has historically been believed that the role of arm motion during walking is related to balancing. Arm motion during natural walking is distinguished in that each arm swing is with the motion of the opposing leg. Although this arm swing motion is generated naturally during bipedal walking, it is interesting to note that the arm swing motion is not necessary for stable walking. This paper attempts to explain the contribution of out-of-phase arm swing in human bipedal walking. Consequently, a human motion control methodology that generates this arm swing motion during walking is proposed. The relationship between arm swing and reaction moment about the vertical axis of the foot is explained in the context of the dynamics of a multi-body articulated system. From this understanding, it is reasoned that arm swing is the result of an effort to reduce the reaction moment about the vertical axis of the foot while the torso and legs are being controlled. This idea is applied to the generation of walking motion. The arm swing motion can be generated, not by designing and tracking joint trajectories of the arms, but by limiting the allowable reaction moment at the foot and minimizing whole-body motion while controlling the lower limbs and torso to follow the designed trajectory. Simulation results, first with the constraint on the foot vertical axis moment and then without, verify the relationship between arm swing and foot reaction moment. These results also demonstrate the use of the dynamic control method in generating arm swing motion. PMID- 18417140 TI - Ultra-performance liquid chromatography/mass spectrometry of intact proteins. AB - Given that numerous small molecule applications of ultra-performance liquid chromatography (UPLC) have been published, efforts were made to examine the potential of UPLC to enhance the separation of intact proteins. Beginning with typically employed conditions, column temperature and organic solvent were optimized followed by an HPLC vs. UPLC comparison. When applied to a mixture of 10 protein standards, the optimized method yielded improved chromatographic resolution, enhanced sensitivity, and a threefold increase in throughput. Subsequent cell lysate analysis demonstrated no compromise in chromatographic or mass spectral data quality at 1/3 of the original run time. PMID- 18417139 TI - The role of hydrogel structure and dynamic loading on chondrocyte gene expression and matrix formation. AB - Crosslinked poly(ethylene glycol) (PEG) hydrogels are attractive scaffolds for cartilage tissue engineering because of their ability to mimic the aqueous environment and mechanical properties of native cartilage. In this study, hydrogel crosslinking density was varied to study the influence of gel structure and the application of dynamic loading (continuous, 1 Hz, 15% amplitude strain) on chondrocyte gene expression over approximately 1 week culture. Gene expression was quantified using real-time RT-PCR for collagen II and aggrecan, the major cartilage extracellular matrix (ECM) components, and collagen I, an indicator of chondrocyte de-differentiation. When chondrocytes were encapsulated in PEG gels with low or high crosslinking, a high collagen II expression compared to collagen I expression (1000 or 100,000:1, respectively) indicated the native chondrocyte phenotype was retained. In the absence of loading, relative gene expression for collagen II and aggrecan was significantly higher (e.g., 2-fold and 4-fold, respectively, day 7) in the low crosslinked gels compared to gels with higher crosslinking. Dynamic loading, however, showed little effect on ECM gene expression in both crosslinked systems. To better understand the cellular environment, ECM production was qualitatively assessed using an in situ immunofluorescent technique and standard histology. A pericellular matrix (PCM) was observed as early as day 3 post-encapsulation and the degree of formation was dependent on gel crosslinking. These results suggest the PCM may protect the cells from sensing the applied loads. This study demonstrates that gel structure has a profound effect on chondrocyte gene expression, while dynamic loading has much less of an effect at early culture times. PMID- 18417141 TI - Calibration of an evaporative light-scattering detector as a mass detector for supercritical fluid chromatography by using uniform Poly(ethylene glycol) oligomers. AB - The quantitativeness of an evaporative light-scattering detector (ELSD) for supercritical fluid chromatography (SFC) was evaluated by using an equimass mixture of uniform poly(ethylene glycol) (PEG) oligomers. Uniform oligomers, in which all molecules have an identical molecular mass, are useful for the accurate calibration of detectors. We calibrated the SFC-ELSD system for various concentrations and molecular masses by using an equimass mixture of PEG oligomers. ELSD not only showed a good linear response to the injected concentration over a wide concentration range, from 10(-4) to 10(-1)g/mL, but also showed a strong dependence on the molecular mass of the solute. By using chromatograms of the equimass mixture of uniform oligomers to calibrate SFC-ELSD, it was possible to determine exact values of not only the average mass but also the molecular-mass distribution for a PEG 1540 sample. The average molecular mass was shifted to a higher value by several percentage points after calibration of the ELSD. PMID- 18417142 TI - Behavior of adsorbed and fluid phases versus retention properties of amino acids on the teicoplanin chiral selector. AB - The relationship between adsorption equilibria of two amino acids, i.e., l,d threonine and l,d-methionine on the teicoplanin chiral selector and their phase behavior has been analyzed. The experimental and numerical methods have been proposed to determine activity coefficients of amino acids in different solvent systems. The procedure was based on the analysis of solubility properties of the amino acids in aqueous solutions of methanol, ethanol and propanol-2-ol used as the mobile phases in chromatographic elution. The solubility measured in mixed alcohol-water solutions was correlated with the non-random-two-liquid (NRTL) model for the activity coefficients. The values of activity coefficients were incorporated into the adsorption isotherm equation, which allowed the analysis of retention properties of the amino acids versus their fluid phase behavior. For the investigation the experimental data of adsorption equilibria of amino acids as well as of the mobile phase constituents acquired in a previous work were exploited. The composition of both the mobile and the adsorbed phases was found to affect the retention properties of the amino acids. For water-rich mobile phases the activity in the adsorbed phase determined the retention mechanism, while for the alcohol-rich systems activity in the mobile phase was predominant. PMID- 18417143 TI - A thermoreversible double gel: characterization of a methylcellulose and kappa carrageenan mixed system in water by SAXS, DSC and rheology. AB - Sol-gel and gel-sol thermal transition of methylcellulose/water, kappa carrageenan/water and methylcellulose/kappa-carrageenan/water mixtures was investigated utilizing small-angle X-ray scattering (SAXS), differential scanning calorimetry (DSC) and oscillatory rheological experiments in temperature regime from 20 to 80 degrees C. Methylcellulose (E461) and kappa-carrageenan (E407) are well-known additives used for gelation in various nutrition and other products. The formulation and characterization of a mixed thermoreversible methylcellulose/kappa-carrageenan/water gel with very interesting double thermal transition gel-sol-gel upon heating was possible. This specific thermal behavior provides a liquid state of the system between the low-temperature and high temperature gel-state and at the same time allows for the easy temperature tuning of the system's state. As such this system is suggested to be further tested as potential carrier for various functional colloidal systems. PMID- 18417144 TI - Physico-chemical investigation of asymmetrical peptidolipidyl-cyclodextrins. AB - A new class of amphiphilic peptidolipidyl-cyclodextrins is reported. The derivatives are chiral due to the presence of an L-leucine in the spacer arm that links a saccharide moiety and a grafted, saturated hydrocarbon chain. Self assembly properties of the peptidolipidyl-cyclodextrins are characterized by quasi-elastic light scattering, turbidity and UV-visible absorption measurements. NMR experiments give insight into the intermolecular dipolar interactions as a function of temperature and concentration. N-dodecyl-N alpha-(6 I-amidosuccinyl-6 1-deoxy-cyclomaltoheptaose)-L-leucine (1) is poorly soluble in aqueous media. N dodecyl-N(alpha)-(6 I-amidosuccinyl-6 I-deoxy-2 I,3 I-di-O-methyl-hexakis-(2 II VII,3 II-VII,6 II-VII-tri-O-methyl)-cyclomaltoheptaose)-L-leucine (2) is found to be more soluble and self-assembles into stable supramolecular colloidal aggregates with nanometric dimensions above a critical aggregation concentration (CAC). It has a propensity for solubilization of hydrophobic species revealing a micellar-like behavior, which is compared to that of the non-ionic detergent octyl glucoside. On the contrary, compound 1 precipitates in a crystalline phase beyond its water solubility limit, and it does not display any solubilizing capacity. The observed behavior corroborates at the molecular level with the NMR results. PMID- 18417145 TI - Glucosamine-carrying temperature- and pH-sensitive microgels: preparation, characterization, and in vitro drug release studies. AB - Glucosamine-carrying temperature- and pH-sensitive microgels with an average diameter of about 100 nm were successfully prepared by free radical precipitation polymerization. The thermo- and pH-responsive properties of the microgels were designed by the incorporation of N-isopropylacrylamide (NIPAM) and acrylic acid (AAc) to copolymerize with acrylamido-2-deoxyglucose (AADG). The stimuli sensitivity of the microgels was studied by the measurement of their sizes and volume phase transition temperature (VPTT) under different surrounding conditions. The results showed that the microgels were responsive to temperature, pH, and ionic strength, and could have a desired VPTT by modifying AADG and AAc contents. The effect of temperature and pH on insulin release from the microgels was also investigated. The release of drug at the tumor-surrounding environment is faster than that under normal physiological conditions. A preliminary in vitro cell study showed that the glucosamine-carrying microgels are more biocompatible to mouse fibroblast cells, compared to the microgels without glucosamine. These glucosamine-carrying dual-sensitive microgels may be promising carriers for targeted drug delivery to tumors. PMID- 18417146 TI - Enhanced fluoride sorption using La(III) incorporated carboxylated chitosan beads. AB - The carboxylated chitosan beads (CCB), which have a defluoridation capacity (DC) of 1385 mg F(-)/kg, have been further chemically modified by incorporating La(3+) ion (La-CCB) and its DC was found to be 4711 mg F(-)/kg whereas the raw chitosan beads (CB) possess only 52 mg F(-)/kg. The fluoride removal by La-CCB is governed by both adsorption and complexation mechanism. The functional groups present in beads were identified by FTIR analysis. The surface condition and existence of fluoride on the beads was confirmed by SEM with EDAX analysis. The experimental data have been analyzed using Freundlich and Langmuir isotherm models. Thermodynamic parameters such as DeltaG(o), DeltaH(o) and DeltaS(o) were calculated to predict the nature of sorption. The kinetic studies were investigated with reaction-based and diffusion-based models. A field trial was carried out with fluoride water collected from a nearby fluoride-endemic village. PMID- 18417147 TI - Interface synthesis of mesoporous MnO2 and its electrochemical capacitive behaviors. AB - Mesoporous MnO(2) has been synthesized by means of a novel, facile, and template free method by virtue of a soft interface between CCl(4) and H(2)O without any surfactants or organometallic precursors or ligands. X-ray diffraction spectroscopy, Fourier transform infrared spectroscopy analysis, scanning electron microscopy, and an ASAP2010 autoadsorption analyzer were applied to investigate the composition and microstructure of the as-synthesized MnO(2). The structure characterizations indicated a good mesoporous structure for as-prepared MnO(2) with an adsorption average pore diameter of 9.7 nm, mesoporous volume of 0.58 cm(3) g(-1), and Brunauer-Emmett-Teller specific surface area of 239 m(2) g(-1). Electrochemical properties of the mesoporous MnO(2) were elucidated by cyclic voltammograms, galvanostatic charge-discharge, and electrochemical impedance spectroscopy in 1 M Na(2)SO(4) electrolyte. Electrochemical data analysis demonstrated that as-synthesized MnO(2) had good capacitive behavior due to its unique mesoporous structure. A specific capacitance of ca. 220 F g(-1) could still be delivered for the mesoporous MnO(2) even at a scan rate of 100 mV s(-1). PMID- 18417148 TI - Film characterization of poly(styrene-butylacrylate-acrylic acid)-silica nanocomposite. AB - To improve the present unsatisfactory UV-shielding and water-resistance properties of many emulsion-type latexes, in this study, poly(styrene butylacrylate-acrylic acid) (PSBA)-grafted-silica (PSBA-g-silica) hybrid nanoparticles were firstly prepared by suspension-dispersion-polymerization. The resulting hybrid nanoparticles were then used to add into PSBA latex to fabricate PSBA nanocomposites. The influence of PSBA-g-silica hybrid nanoparticles on the interfacial adhesion, film morphology, thermodynamics properties, UV-shielding, water-resistance and mechanical performance of PSBA films were investigated systematically. Our results revealed that filling 1.5 wt% PSBA-g-silica hybrid nanoparticles resulted in significant improvement in interfacial adhesion properties, UV-shielding, water-resistance and mechanical properties. Fabricating the silica-based latex nanocomposite is an effective approach to develop the new emulsion-type paints and adhesives. PMID- 18417149 TI - Donnan potential and surface potential of a spherical soft particle in an electrolyte solution. AB - A simple numerical method, which does not involve numerical integration of the Poisson-Boltzmann equations, is presented for obtaining the relationship between the Donnan potential and surface potential of a spherical soft particle (i.e., a polyelectrolyte-coated particle) in a symmetrical electrolyte solution. We assume that a soft particle consists of the particle core of radius a covered with an ion-penetrable surface layer of polyelectrolytes of thickness d and that ionized groups of valence Z are distributed at a uniform density of N in the polyelectrolyte layer and the relative permittivity takes the same value in the regions outside and inside the polyelectrolyte layer. The Donnan potential and surface potential are determined by the values of a, d, Z, N, and the Debye Huckel parameter kappa of the electrolyte solution. Numerical results obtained by the present method are in excellent agreement with exact results obtained by solving the nonlinear spherical Poisson-Boltzmann equations for the both regions inside and outside the polyelectrolyte layer. PMID- 18417150 TI - Simple and highly sensitive assay system for TNFR2-mediated soluble- and transmembrane-TNF activity. AB - Drugs that target tumor necrosis factor-alpha (TNF) are particularly important in the treatment of severe inflammatory progression in rheumatoid arthritis, Crohn's disease and psoriasis. Despite the central role of the TNF/TNF receptor (TNFR) in various disease states, there is a paucity of information concerning TNFR2 signaling. In this study, we have developed a simple and highly sensitive cell death based assay system for analyzing TNFR2-mediated bioactivity that can be used to screen for TNFR2-selective drugs. Using a lentiviral vector, a chimeric receptor was engineered from the extracellular and transmembrane domain of human TNFR2 and the intracellular domain of mouse Fas and the recombinant protein was then expressed in TNFR1(-/-)R2(-/-) mouse preadipocytes. Our results demonstrate that this chimeric receptor is capable of inducing apoptosis by transmembrane- as well as soluble-TNF stimuli. Moreover, we found that our bioassay based on cell death phenotype had an approximately 80-fold higher sensitivity over existing bioassays. We believe our assay system will be an invaluable research tool for studying TNFR2 and for screening TNFR2-targeted drugs. PMID- 18417151 TI - Comparison of carboxylated and Penta-His microspheres for semi-quantitative measurement of antibody responses to His-tagged proteins. AB - The Luminex system is a flow cytometry based tool that permits the simultaneous measurement of many analytes from just a single serum sample. The technology uses microspheres, which are available in different colors and can be coated with different kinds of biomolecules. For the immobilisation of His-tagged proteins, two types of beads can be used: chemically activated carboxylated beads or Penta His beads, which have antibodies against His-tags on their surface. In this study, we compared carboxylated and Penta-His beads. For carboxylated as compared to Penta-His beads, the non-specific background is lower (Median Fluorescence Intensity; MFI>250, 0% versus 15%), the specific signal intensity is higher (mean MFI 2860 versus 722) and not dependent on the configuration of the protein. Above all, the protein coupled carboxylated beads are useful over longer periods of time. Therefore, we conclude that for developing a multiplex assay for semi quantitative measurement of antibody responses against His-tagged proteins the best microspheres to use are the carboxylated ones. PMID- 18417152 TI - The adverse cardiopulmonary phenotype of caveolin-1 deficient mice is mediated by a dysfunctional endothelium. AB - Recently generated caveolin-1 deficient mice (cav-1(-/-)) display several physiological alterations such as severe heart failure and lung fibrosis. The molecular mechanisms how the loss of caveolin-1 (cav-1) mediates these alterations are currently under debate. A plethora of studies support a role of cav-1 as a negative regulator of endothelial nitric oxide synthase (eNOS). Accordingly, constitutive eNOS hyperactivation was observed in cav-1(-/-). Given the hyperactivated eNOS enzyme we hypothesized that disturbed eNOS function is involved in the development of the cardiopulmonary pathologies in cav-1(-/-). The present study argues that loss of cav-1 results in enhanced eNOS activity but not in increased vascular tetrahydrobiopterin (BH(4)) levels (which acts as an essential eNOS cofactor) thereby causing a stoichiometric discordance between eNOS activity and BH(4) sufficient to cause dysfunctional eNOS signaling. The resultant oxidative stress is largely responsible for major cardiac and pulmonary defects observed in cav-1(-/-). BH(4) donation to cav-1(-/-) led to a normalized BH(4)/BH(2) ratio, to reduced oxidant stress, to substantial improvements of both systolic and diastolic heart function and to marked amelioration of the impaired lung phenotype. Notably, the antioxidant tetrahydroneopterin which is not essential for eNOS function showed no relevant effect. Taken together these novel findings indicate that dysfunctional eNOS is of central importance in the genesis of the cardiopulmonary phenotype of cav-1(-/-). Additionally, these findings are generally of paramount importance since they underline the deleterious role of an uncoupled eNOS in cardiovascular pathology and they additionally suggest BH(4) as an effective cure. PMID- 18417154 TI - Chaotic gene regulatory networks can be robust against mutations and noise. AB - Robustness to mutations and noise has been shown to evolve through stabilizing selection for optimal phenotypes in model gene regulatory networks. The ability to evolve robust mutants is known to depend on the network architecture. How do the dynamical properties and state-space structures of networks with high and low robustness differ? Does selection operate on the global dynamical behavior of the networks? What kind of state-space structures are favored by selection? We provide damage propagation analysis and an extensive statistical analysis of state spaces of these model networks to show that the change in their dynamical properties due to stabilizing selection for optimal phenotypes is minor. Most notably, the networks that are most robust to both mutations and noise are highly chaotic. Certain properties of chaotic networks, such as being able to produce large attractor basins, can be useful for maintaining a stable gene-expression pattern. Our findings indicate that conventional measures of stability, such as damage propagation, do not provide much information about robustness to mutations or noise in model gene regulatory networks. PMID- 18417153 TI - An evaluation of anxiety sensitivity, emotional dysregulation, and negative affectivity among daily cigarette smokers: relation to smoking motives and barriers to quitting. AB - The present investigation evaluated the relations between anxiety sensitivity and motivational bases of cigarette smoking, as well as barriers to quitting smoking, above and beyond concurrent substance use, negative affectivity, and emotional dysregulation among a community sample of 189 daily cigarette smokers (46% women; M(age)=24.97 years, SD=9.78). Results indicated that anxiety sensitivity was significantly related to coping, addictive, and habitual smoking motives, as well as greater perceived barriers to quitting. These effects were evident above and beyond the variance accounted for by concurrent tobacco, alcohol, and marijuana use and discernable from shared variance with negative affectivity and emotional dysregulation. Emotional dysregulation was significantly related to stimulation, habitual, and sensorimotor smoking motives and greater perceived barriers to quitting, whereas negative affectivity was only significantly related to smoking for relaxation. These findings uniquely add to a growing literature suggesting anxiety sensitivity is an important and unique cognitive factor for better understanding clinically-relevant psychological processes related to cigarette smoking. PMID- 18417155 TI - The establishment of a novel non-alcoholic steatohepatitis model accompanied with obesity and insulin resistance in mice. AB - Non-alcoholic steatohepatitis (NASH) is a hepatic manifestation of the metabolic syndrome that can progress to liver cirrhosis. The major aim of this study was to establish a novel NASH mouse model accompanied by obesity and insulin resistance, then explore the molecular mechanisms of NASH and evaluate the effects of both the peroxisome proliferator-activated receptor alpha (PPARalpha) agonist fenofibrate and the PPARgamma agonist rosiglitazone in this established NASH model. The novel model was induced in C57BL/6 mice by 23 weeks of ad libitum feeding of a modified high-fat diet (mHFD), with lower methinione and choline and higher fat content. In comparison to the controls, the model animals developed pronounced obesity, dyslipidemia and insulin resistance. Marked liver lesions characterized by severe steatosis, inflammation, fibrosis, increased hepatic triglyceride content, and elevated serum alanine aminotransferase (ALT) levels were observed in the models. In this novel model, treatment with fenofibrate or rosiglitazone significantly improved insulin sensitivity and corrected dyslipidemia; however, fenofibrate was more effective than rosiglitazone in improving hepatic morphology and ALT levels. Further study showed that long-term feeding of mHFD significantly increased expression of mRNA for hepatic PPARgamma, adipose fatty acid binding protein (ap2) and CD36 and suppressed expression of mRNA for hepatic PPARalpha and carnitine palmitoyl transferase-1a (CPT-1a). These results showed the successful establishment of the combined NASH and obese insulin resistance mouse model. Additionally, aberrant expressions of hepatic PPARalpha and PPARgamma may play a major role in the pathogenesis of NASH by affecting hepatic lipogenesis and fatty acid oxidation in this novel model. PMID- 18417157 TI - [Epilepsy and insula]. AB - The insula is the only cortical part of the brain that is not visible on the surface of the hemisphere, because it is totally covered by the frontoparietal and temporal opercula. The insula is triangular in shape and is separated from the opercula by the anterior, superior, and inferior peri-insular sulci. It is morphologically divided into two parts by the central insular sulcus. The anterior part of the insula bears three short gyri, and its posterior part contains two long gyri. The vascular supply of the insula is mainly provided by the M2 segment of the middle cerebral artery, a substantial obstacle to any open or stereotactic procedure aiming at the insular region. The insula is functionally involved in cardiac rhythm and arterial blood pressure control, as well as in visceromotor control and in viscerosensitive functions. There is substantial evidence that the insula is involved as a somesthetic area, including a major role in the processing of nociceptive input. The role of the insula in some epilepsies was recently investigated by means of depth electrode recordings made following Talairach's stereoelectroencephalography (SEEG) methodology. It appears that ictal signs associated with an insular discharge are very similar to those usually attributed to mesial temporal lobe seizures. Ictal symptoms associated with insular discharges are mainly made up of respiratory, viscerosensitive (chest or abdominal constriction), or oroalimentary (chewing or swallowing) manifestations. Unpleasant somatosensory manifestations, always opposite the discharging side, are also frequent. Ictal signs arising from the insula occur in full consciousness; these are always simple partial seizures. Seizures arising from the temporal lobe always invade the insular region, but in approximately 10% of cases, the seizures originate in the insular cortex itself. These data explain that there has been a rebirth of interest in the insula from a surgical perspective over the past few years. The literature contains no reports of cases of resection of insular cortex alone; most insular resections are performed in the context of temporal resection, when there is some evidence of seizures originating in the insula itself. Such procedures are risky and their efficacy, in terms of postoperative surgical outcome, has not yet been clearly assessed. In this context, less invasive procedures, such as SEEG-guided radiofrequency thermolesions of the insular cortex, are under investigation. PMID- 18417158 TI - [Multiple SEEG-guided RF-thermolesions of epileptogenic foci]. AB - In many patients with drug-resistant partial epilepsy, depth electrode recordings may be required to delineate the best region for cortical resection. We usually implant depth electrodes according to Talairach's stereoelectroencephalography (SEEG) methodology. Using these chronically-implanted depth electrodes, it is possible to generate radiofrequency (RF) thermolesions of the epileptic foci and networks. The advantages of this type of technique are supported by several lines of evidence, in particular, the high number of implanted electrodes makes it possible to generate several thermolesions, whereas the bleeding risk is null, since no additional electrode trajectory is required. Lesions are generated using 100- to 120-mA bipolar current (50V), applied for 10-40s within the epileptogenic zone, as identified by the SEEG recordings. No general or neurological complication occurred during the procedures. Forty-three patients investigated with video-SEEG recordings for presurgical assessment of drug-resistant partial epilepsy were treated using SEEG-guided RF-thermolesions of the epileptic foci between 2001 and 2006, with a follow-up ranging from 12 to 66 months. Three patients were seizure-free and 52% of the patients had a decrease in their seizure frequency of at least 50%. Of the patients presenting a malformation of cortical development etiology (i.e. dysplasia or heterotopia), 70% were classified as responders (at least a 50% decrease in seizure frequency) (p=0.052), whereas the results were less favorable in patients with a cryptogenic and hippocampal sclerosis etiology. Twenty patients underwent conventional cortectomy in a second step, 18 of whom are in Engel class I. In conclusion, SEEG guided RF-thermolesions of the epileptic foci and networks proved to be a safe therapeutic procedure capable of providing an immediate benefit in terms of seizure control, especially in patients with epilepsy symptomatic of cortical development malformation. Such thermolesions do not preclude subsequent conventional surgery in case of failure, which can be proposed as an alternative procedure if no resective surgery is possible. PMID- 18417156 TI - Microfibril-associate glycoprotein-2 (MAGP-2) promotes angiogenic cell sprouting by blocking notch signaling in endothelial cells. AB - Angiogenesis is highly sensitive to the composition of the vascular microenvironment, however, our understanding of the structural and matricellular components of the vascular microenvironment that regulate angiogenesis and the molecular mechanisms by which these molecules function remains incomplete. Our previous results described a novel pro-angiogenic activity for Microfibril Associated Glycoprotein-2 (MAGP-2), but did not address the molecular mechanism(s) by which this is accomplished. We now demonstrate that MAGP-2 promotes angiogenic cell sprouting by antagonizing Notch signaling pathways in endothelial cells. MAGP-2 decreased basal and Jagged1 induced expression from the Notch sensitive Hes-1 promoter in ECs, and blocked Jagged1 stimulated Notch1 receptor processing in transiently transfected 293T cells. Interestingly, inhibition of Notch signaling by MAGP-2 seems to be restricted to ECs since MAGP 2 increased Hes-1 promoter activity and Notch1 receptor processing in heterologous cell types. Importantly, constitutive activation of the Notch signaling pathway blocked the ability of MAGP-2 to promote angiogenic cell sprouting, as well as morphological changes associated with angiogenesis. Collectively, these observations indicate that MAGP-2 promotes angiogenic cell spouting in vitro by antagonizing Notch signaling pathways in ECs. PMID- 18417159 TI - [Epilepsy surgery in children: from a multidisciplinary network to the creation of an expertise and care centre]. AB - We report on the experience of a network created in 1994 to evaluate children with drug-resistant epilepsies who are candidates for surgical treatment. The network includes epilepsy units from several university hospitals in France that decided to share not only their clinical expertise to better respond to the need for a multidisciplinary approach of epilepsy surgery in children, but also all the technical and human resources available in the various teams. This mode of operation has certainly provided concrete proof of its efficacy since it undoubtedly facilitated, and even accelerated, access to optimal presurgical evaluation and epilepsy surgery for hundreds of children. However, after 10 years of this very enriching practice it became evident that our approach was certainly necessary but not sufficient. It is estimated that every year in France nearly 500 children are candidates for surgical treatment, and following a presurgical evaluation, 50% of them could be operated on. Today, only 150-200 children have access to a presurgical evaluation every year. This is a highly paradoxical situation since, even if the human suffering component that such a situation generates is set aside, the direct and indirect life-time costs for every 100 nonoperated patients is estimated at 40 million euros. As a result of our cumulated experience, in 2004 we proposed a different operating model with the creation of an expertise center that will combine not only medical care services provided by a fully equipped multidisciplinary team, but also a pole of applied clinical and fundamental research, a medicosocial center managed by a lay association and an industrial development pole. The project has been recently validated by the Ministry of Health and is supported by a number of national and regional institutions. The Institute for Children and Adolescents with Epilepsy- IDEE--is designed to accelerate diagnostic procedures and, when indicated, access to optimal presurgical evaluation, while also serving as a model for a medical and economic evaluation of epilepsy care in children. PMID- 18417160 TI - [Cellular mechanisms of the epilepsies: In vitro studies on human tissue]. AB - BACKGROUND AND PURPOSE: Animal models have provided very valuable data to specify the physiopathological mechanisms of the various forms of epilepsy. However, the question arises of knowing which of these experimental results are relevant to the human epileptic brain. The development of epileptic surgery makes it possible to directly study the functional properties of human brain tissue in vitro and to analyze the mechanisms underlying seizures and epileptogenesis. We review some of the results obtained over the last few years in our laboratory based on electrophysiological, immunocytochemical and molecular experiments conducted on human brain tissue. RESULTS: This review covers a number of the mechanisms of neuronal synchronizations generating epileptiform discharges, including the role of electrical synapses connecting the inhibitory interneurons, particularly in Taylor-type focal cortical dysplasia and the functional lability of GABAergic inhibition in epileptogenic human cortical tissue, which may sustain triggering and propagation of seizures. Some of these mechanisms have not been described in animal models. CONCLUSIONS: Studies on human tissue, when carefully designed, are necessary to validate the data collected on animal models and will continue to provide us with new and important information on the cerebral changes related to epilepsy. Moreover, these studies allow development of a class of antiepileptic drugs that have a completely new mechanism of action, which could be effective in the treatment of drug-resistant epilepsies. PMID- 18417161 TI - [Can functional MRI replace the Wada test?]. AB - Temporal lobe epilepsy (TLE) is usually associated with specific impairment of memory functions. A reliable assessment of memory function is therefore a crucial element of the presurgical evaluation for epilepsy surgery, especially for temporal lobectomy. To date, intracarotid amobarbital testing remains the gold standard in evaluating memory before planning surgery in patients with refractory TLE, when neuropsychological testing is not contributive. Functional MRI holds great promise as a powerful tool in memory evaluation. Preliminary studies in epileptic patients have shown promising results. In the future, functional MRI will certainly contribute to partially replacing the Wada test or to evaluating the hippocampal memory capacities independently. PMID- 18417162 TI - [High-resolution EEG (HR-EEG) and magnetoencephalography (MEG)]. AB - HR-EEG (high resolution EEG) and MEG (magnetoencephalography) allow the recording of cerebral electromagnetic activities with excellent temporal resolution. These tools have also considerably progressed in spatial resolution and now constitute real methods of Electric and Magnetic Source Imaging. Their limits and the precision of the results obtained are discussed in distinct types of partial epilepsy. HR-EEG and MEG allow localization of scalp-EEG interictal spikes and more rarely ictal activities. They now contribute to the presurgical evaluation of pharmacoresistant partial epilepsies. These investigations appear to be of particular importance in presurgical assessment of MRI-negative epilepsy. PMID- 18417163 TI - [Temporal disconnection as an alternative treatment for intractable temporal lobe epilepsy: techniques, complications and results]. AB - Temporal lobe epilepsy (TLE) is the most common form of intractable partial epilepsy in adults. Surgery (lobectomy or amygdalohippocampectomy) is effective in most patients. However, some complications can occur and brain shift, hematoma into the post operative cavity and occulomotor nerve palsy have been reported due to the surgical technic. We report the technique, safety and efficacy of temporal disconnection in nonlesional TLE. Forty-seven patients (18 males, 29 females; handedness: 12 left, 33 right; aged 35 years+/-10; mean duration of epilepsy: 24+/-10 years) underwent temporal disconnection (20 left, 27 right) guided by neuronavigation. Sixteen patients (35 %) underwent additional presurgical evaluation with SEEG. The outcome was assessed using Engel's classification. At the two-year follow-up, 85 % of the patients were seizure-free (Engel I), 26 (58 %) of whom were Ia. Postoperative persistent morbidity included mild hemiparesis (n=1), mild facial paresis (n=1), quadranopsia (n=23) and hemianopia (n=1). Verbal memory worsened in 13 % of cases when the disconnection was performed in the dominant lobe. MRI follow-up showed two cases of nonsymptomatic thalamic or pallidal limited ischemias, two cases of temporal horn-cystic dilatation, one requiring surgical reintervention without sequelae. There was one case of postoperative phlebitis. In the seizure-free patient group, postoperative EEG showed interictal temporal spikes at three months, one year and two years located in the anterior temporal region. Temporal disconnection is effective, prevents the occurrence of subdural cyst and hematomas in the temporal cavity, prevents the occurrence of oculomotor palsy, and limits the occurrence of quadranopsia. However, comparative studies are required to evaluate temporal disconnection as an alternative to lobectomy in nonlesional TLE. PMID- 18417164 TI - [Ictal single photon computed tomography and SISCOM: methods and utility]. AB - Ictal single photon emission computed tomography (SPECT) reflects epileptic activity through hyperperfusion associated with ictal discharge. It provides valuable spatial information on ictal activity, but its temporal resolution is limited. Therefore, information provided by SPECT is not restricted to the epileptogenic zone, but demonstrates a larger epileptogenic network, related to the spatiotemporal dynamics of ictal processes. This review includes a description of the technique, followed by a description of the different parameters likely to influence the ictal perfusion images. SPECT gives contributes original diagnostic data to the decision-making process which will complete, the other evaluation parameters. PMID- 18417165 TI - [Safety and efficacy of multiple subpial transections: report of a consecutive series of 30 cases]. AB - PURPOSE: To present our results using multiple subpial transections (MST) for the treatment of pharmacologically refractory epilepsy (PRE) with epileptogenic foci in eloquent areas. METHOD: Between January 2003 and March 2006, we treated 33 patients with PRE with epileptogenic foci in eloquent areas by MST "in rays", either isolated (MSTs group) or completing resection or disconnection of other cortical areas (MST+ group). Our first 30 patients had a follow-up of at least 24 months: eight in the MSTs group and 22 in the MST+ group. Four postoperative grades were distinguished based on a modified Engel classification: seizure-free (100% seizure reduction equals to Grade I), substantial significant seizure reduction (75% to 99% seizure reduction equals to Grade II), moderate significant reduction (50% to 74% seizure reduction equals to Grade III) and finally no significant reduction (seizure reduction less than 50% equals to Grade IV). RESULTS: In the MSTs group, two patients (25%) were in grade I and five (62%) in grade II or III. In the MST+ group, six patients (27%) were in grade I and 13 (59%) in grade II or III. All patients showed some seizure reduction and some improvement in behavior or cognitive function with no permanent neurological deficit. CONCLUSION: This series supports the notion that multiple subpial transections are associated with a significant seizure reduction (in 86.6% of the cases reported herein) and that the risk of permanent neurological deficit can be very low. PMID- 18417166 TI - [Radiosurgery for drug-resistant epilepsies: state of the art, results and perspectives]. AB - BACKGROUND: There is growing interest in the use of radiosurgery in epilepsy. We analyzed our experience in this field in an attempt to define the potential of radiosurgery in epileptology. MATERIAL AND METHODS: [corrected] Our local clinical experience (134 patients), accumulated over the last 15 years, mainly includes treatment of temporal lobe epilepsy without space-occupying lesions (59 patients), including 53 with pure MTLE, 61 cases of hypothalamic hamartoma, two cases of callosotomy, and 12 other types of epilepsy. RESULTS: The analysis of our material, as well as other clinical and experimental data, suggest that the use of radiosurgery is beneficial only to patients in whom a strict preoperative definition of the extent of the epileptogenic zone (or network) has been achieved and strict rules of dose planning have been applied. As soon as these principles are not observed, the risk of treatment failure and/or side effects increases dramatically. Long-term outcome data are now available and published for MTLE but not yet for other types of epilepsy. Long-term safety and efficacy in MTLE are comparable to surgical resection but radiosurgery has the advantage of sparing verbal memory in patients operated by Gamma Knife (GK) on the dominant side. In small hamartomas, the efficacy is comparable to microsurgery but with a dramatic reduction in risk. CONCLUSION: The vast amount of clinical materiel and long-term evaluation now support the use of GK surgery in small hypothalamic hamartomas and MTLE when the patient is at risk of verbal memory loss. PMID- 18417167 TI - [Animal models to develop surgery of focal epilepsies?]. AB - A model is a simplified preparation that reproduces only the most critical features of a disease. To be considered as a validated animal model, such an experimental preparation must fulfill three criteria: isomorphism or similarities of the symptoms; predictivity or identical pharmacological reactivity; homology or etiological similarity. In epilepsy, the use of animal models helps our understanding of physiological and pathological networks involved in the genesis, maintenance, and propagation of seizures. The animal models of epilepsy are also useful in designing and testing new surgical therapeutical strategies, in particular using deconnection or neuromodulation in drug-resistant focal epilepsies. Here we describe three animal models of focal epilepsy, adapted to addressing experimental surgery issues. Kindling consists in the regular liminar stimulation of a given brain structure in the rodent to develop a focal discharge that is secondarily generalized. The local application of epileptogenic agents such as cobalt, iron, or penicillin leads to focal discharges that do not generalize in the rodent or the primate. It is a model of focal neocortical epilepsy without secondary generalization. The focal application of kainate, an excitotoxic glutamate agonist, in the dorsal hippocampus of the adult mouse results, after a latent period, in spontaneous and recurrent focal discharges, behavioral interictal troubles, drug resistance, and histological anomalies reminiscent of hippocampal sclerosis. This constitutes a model of mesial-temporal epilepsy. Better knowledge, in these models, of the neural networks generating, propagating, and/or controlling the seizures should make it possible to design innovative surgical approaches for the treatment of drug-resistant epilepsies. PMID- 18417168 TI - [Changes in spontaneous epileptic activity after selective intrahippocampal transection in a model of chronic mesial temporal lobe epilepsy]. AB - Drug-resistant mesial temporal lobe epilepsy with hippocampal sclerosis is associated with anatomical, ultrastructural and functional changes that facilitate generation and spread of epileptic seizures. Intrahippocampal circuits are modified in their transversal lamellar and longitudinal translamellar organization. Neuronal death and the neuroplasticity of surviving cells contribute to major phenomena: an increased hyperexcitability of the hippocampal formation and an increased synchronization of its principal cells. Selective disruption of the epileptic networks that are involved in mesial temporal lobe epilepsy may have a therapeutic effect. We present here the preliminary results of a selective intrahippocampal transection in a chronic model of mesial temporal lobe epilepsy after focal injection of kainic acid in adult mice. A complete transection of the hippocampal formation (including dentate gyrus and hippocampus proper, sparing the fimbria) results in a blockade of ictal activities spread from the generator, a reduction in their frequency and an increase in their duration. In contrast, after a transection sparing the dentate gyrus and hilus, no modification was noted. In this model of mesial temporal lobe epilepsy, longitudinally projecting axonal circuits of the dentate gyrus and hilus appear to be implicated in generation, propagation and interruption of ictal activities within hippocampal formation. PMID- 18417169 TI - [Neuropsychological assessment in epilepsy surgery of children]. AB - In children with epilepsy, surgical treatment has been recognized as a valuable therapeutic option that can improve the cognitive prognosis and facilitate schooling. Most of the earlier studies focused on general intellectual outcome and behavior. However, substantial progress and new developments in the neuropsychological evaluation of epileptic children treated with surgery have been made over the past few years. Several recent studies explored language, memory, executive functions and behavior in patients undergoing surgery, taking into account cerebral maturation and plasticity processes. Neuropsychological studies have demonstrated greater functional recovery after surgery in children than in adults. Notably, there is evidence of particular reorganization processes for language dominance. Other studies demonstrate that early surgery of temporal lobe epilepsy in children is associated with better memory than in adults. Longitudinal studies are still needed to describe developmental trajectories and optimize neuropsychological rehabilitation. PMID- 18417170 TI - [Drug resistance in partial epilepsy: epidemiology, mechanisms, pharmacogenetics and therapeutical aspects]. AB - It has been established that 20-30% of epilepsies are not controlled by antiepileptic drugs. Drug resistance is associated with several major problems, including prognosis, cognitive function, behavior, mortality, cost and quality of life. Apart from classic risk factors for drug resistance, such as neurological, psychiatric, imaging, EEG abnormalities, a high frequency of seizures before medical therapy and complex febrile convulsions, the potential role of multidrug transporters as well as their genetic control and the altered sensitivity of neuronal drug receptors has gained growing attention. In the future, pharmaceutical engineering may bypass these factors. To a certain extent, drug resistance may develop progressively in a neurobiological process and the control of this process could limit its development. PMID- 18417171 TI - [Dual pathology]. AB - Dual pathology is defined as the association of two potentially epileptogenic lesions, hippocampal (sclerosis, neuronal loss) and extrahippocampal (temporal or extratemporal). Epileptic activity may be generated by either lesion and the relative importance of every lesion's epileptogenicity conditions the surgical strategy adopted. Most frequently associated with hippocampal sclerosis are cortical dysplasias. The common physiopathology of the two lesions is not clearly established. Extrahippocampal lesions may be undetectable on MRI (microdysgenesis, for example) and ictal discharge patterns may vary among dual pathology patients. The surgical strategy depends on the location of the extrahippocampal lesion and its relative role in seizure generation; however, reported surgical results suggest that simultaneous resection of mesial temporal structures along with the extrahippocampal lesion should be performed. PMID- 18417172 TI - [Relevance of basic research to clinical data: Good answers, wrong questions!]. AB - What conclusions can be derived from experimental data on human epilepsies? This review discusses these issues, notably concerning human temporal lobe epilepsies (TLEs) and infantile epilepsies, where important advances have been achieved in both theory and the comprehension of epileptogenic mechanisms. A wide spectrum of human and animal data converge to show that the naive network transforms to one that generates seizures spontaneously. Thus, in TLE, experimental and human data suggest that the inaugurating status generates a sequence of events that lead to the sprouting of fibers and the formation of novel excitatory synapses. This reactive plasticity constitutes a basis for the generation of novel seizures by the epileptic network. Similarly, in vitro studies indicate that in immature hippocampal formation, the propagation of high- but not low-frequency seizures can transform a naive network into one that generates further seizures, thereby, giving an indication as to the types of seizure that are epileptogenic. In conclusion, it is suggested that although animal data cannot mimic human seizures in all their complex and variable etiologies, it provides essential indications on the mechanisms that enable seizure generation. PMID- 18417173 TI - Abolished thermal and mechanical antinociception but retained visceral chemical antinociception induced by butorphanol in mu-opioid receptor knockout mice. AB - Butorphanol is hypothesized to induce analgesia via opioid pathways, although the precise mechanisms for its effects remain unknown. In this study, we investigated the role of the mu-opioid receptor (MOP) in thermal, mechanical, and visceral chemical antinociception induced by butorphanol using MOP knockout (KO) mice. Butorphanol-induced thermal antinociception, assessed by the hot-plate and tail flick tests, was significantly reduced in heterozygous and abolished in homozygous MOP-KO mice compared with wildtype mice. The results obtained from our butorphanol-induced mechanical antinociception experiments, assessed by the Randall-Selitto test, were similar to the results obtained from the thermal antinociception experiments in these mice. Interestingly, however, butorphanol retained its ability to induce significant visceral chemical antinociception, assessed by the writhing test, in homozygous MOP-KO mice. The butorphanol-induced visceral chemical antinociception that was retained in homozygous MOP-KO mice was completely blocked by pretreatment with nor-binaltorphimine, a kappa-opioid receptor (KOP) antagonist. In vitro binding and cyclic adenosine monophosphate assays also showed that butorphanol possessed higher affinity for KOPs and MOPs than for delta-opioid receptors. These results molecular pharmacologically confirmed previous studies implicating MOPs, and partially KOPs, in mediating butorphanol-induced analgesia. PMID- 18417174 TI - Psychosocial stress and chronic ethanol ingestion in male rats: effects on elevated plus maze behavior and ultrasonic vocalizations. AB - Psychosocial stress is known to enhance anxiety levels in rodents. The present study evaluated the effect of long-term ingestion of ethanol (EtOH) on anxiety levels of male Long Evans rats housed either singly or in triads (three/cage). Based on measures of offensive and defensive behaviors, triad-housed rats were designated as being dominant, subdominant or subordinate. The effect of chronic ingestion of a 6% EtOH solution was assessed on behaviors in the elevated plus maze (EPM) test, and on air-puffs elicited 22 kHz ultrasonic vocalizations (USV). EtOH naive dominant rats showed less anxiogenic-like (open arms time) and displacement behavior (grooming), but more risk assessment behavior (stretch attend postures) than their cage-mates or the single-housed rats. Intake of EtOH prior to the EPM test was lower in dominant and single-housed rats compared to subdominant and subordinate rats. Triad-housed rats, but not the single-housed rats, decreased their intake of EtOH after the EPM test. Overall, EtOH intake had an anxiolytic effect in all rats, but the effect was most prominent in single housed rats. Furthermore, EtOH intake decreased air-puff induced USVs of triad housed rats, but increased USVs of single-housed rats. These data indicate that housing condition and rank status influence the emotional state of male rats. Furthermore, the anxiolytic effect of voluntarily ingested EtOH was depended on these variables. PMID- 18417175 TI - Limonoids from the stem bark of Cedrela odorata. AB - Four nomilin/obacunol derivatives and a swietenolide derivative, together with seven known limonoids, were isolated from stem bark of Cedrela odorata and their structures established by spectroscopic methods. Antifeedant activity of the isolated compounds was also tested. PMID- 18417176 TI - Screening of chemical composition, antimicrobial and antioxidant activities of Artemisia essential oils. AB - The chemical composition of essential oils isolated from aerial parts of seven wild sages from Western Canada -Artemisia absinthium L., Artemisia biennis Willd., Artemisia cana Pursh, Artemisia dracunculus L., Artemisia frigida Willd., Artemisia longifolia Nutt. and Artemisia ludoviciana Nutt., was investigated by GC-MS. A total of 110 components were identified accounting for 71.0-98.8% of the oil composition. High contents of 1,8-cineole (21.5-27.6%) and camphor (15.9 37.3%) were found in Artemisia cana, A. frigida, A. longifolia and A. ludoviciana oils. The oil of A. ludoviciana was also characterized by a high content of oxygenated sesquiterpenes with a 5-ethenyltetrahydro-5-methyl-2-furanyl moiety, of which davanone (11.5%) was the main component identified. A. absinthium oil was characterized by high amounts of myrcene (10.8%), trans-thujone (10.1%) and trans-sabinyl acetate (26.4%). A. biennis yielded an oil rich in (Z)-beta-ocimene (34.7%), (E)-beta-farnesene (40.0%) and the acetylenes (11.0%) (Z)- and (E)-en-yn dicycloethers. A. dracunculus oil contained predominantly phenylpropanoids such as methyl chavicol (16.2%) and methyl eugenol (35.8%). Artemisia oils had inhibitory effects on the growth of bacteria (Escherichia coli, Staphylococcus aureus, and Staphylococcus epidermidis), yeasts (Candida albicans, Cryptococcus neoformans), dermatophytes (Trichophyton rubrum, Microsporum canis, and Microsporum gypseum), Fonsecaea pedrosoi and Aspergillus niger. A. biennis oil was the most active against dermatophytes, Cryptococcus neoformans, Fonsecaea pedrosoi and Aspergillus niger, and A. absinthium oil the most active against Staphylococcus strains. In addition, antioxidant (beta-carotene/linoleate model) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activities were determined, and weak activities were found for these oils. PMID- 18417177 TI - Rescue chemotherapy for patients with resistant or relapsed endemic Burkitt's lymphoma. AB - Patients with endemic Burkitt's lymphoma who failed primary treatment with the Malawi 2002 or 2003 Burkitt's lymphoma treatment protocols, consisting of high frequency cyclophosphamide 40 mg/kg and intrathecal methotrexate, were offered rescue chemotherapy. Twenty-eight patients (14 boys and 14 girls; age range 3-13 years) with resistant disease (n=8) or relapse (n=20) presented to the Queen Elizabeth Central Hospital, Blantyre, Malawi. Treatment consisted of cyclophosphamide 60 mg/kg and vincristine 1.5 mg/m(2) i.v. on Days 1, 8 and 15, plus intrathecal methotrexate on the same days in those patients treated for a relapse. The majority of patients (81%) had St Jude stage III or IV disease. Twenty patients (71%) achieved a complete clinical remission. Day 8 treatment was delayed in eight children and Day 15 treatment in five patients, both for a median of 7 days, mainly due to neutropenia. Ten patients relapsed after 42-311 days (median 105 days). Ten patients (36%) remained in remission for 353-712 days (median 487 days). Patients whose first relapse occurred after 6 months as well as those with limited disease had the best outcome. This simple 15-day chemotherapy schedule salvaged 36% of patients and significantly increased the overall cure rate of our Burkitt's lymphoma patients. PMID- 18417178 TI - Collection and evaluation of semen from the three-toed sloth (Bradypus tridactylus). AB - Sloths (Bradypus sp.) are extremely sensitive animals that suffer with the destruction and fragmentation of forests. They present a low population growth rate and need to be further studied for the preservation of the specie. Thus, the aim of this study was to establish an efficient semen collection protocol as well as characterize sperm concentration, motility and morphology in order to contribute with information about the reproductive traits of this specie, which has never been described in the literature before. For that, nine Bradypus tridactylus males were captured during the wet season and six during the dry season, in Manaus (AM), located in the north region of Brazil, semen was collected by electroejaculation with shocks given in sequences of progressive intensities (minimum 20mA and maximum 60mA). All animals ejaculated small volumes of semen and in some of them, the volume ejaculated was not enough for a complete spermiogram. Physical characteristics observed on the collections of the wet season were different from those seen in the specimen collected in the dry season. Motility an vigor was very low and did not show forward progression, only oscillatory movement. After Spermac stain, spermatozoa presented a wide variety of defects; however, the differences in morphology were not significant between seasons. The morphology assessed by scanning electron microscopy shows that the head in both groups could be elongated, short or could have a base narrower than the apex and the midpiece narrowed abruptly, forming a nip in its transition to the tail. Although further studies are necessary to verify our preliminary findings concerning seasonal variation in sperm quality, these results demonstrate that semen can be safely collected from sloths by electroejaculation and provide the first reports of semen characteristics in this species. PMID- 18417179 TI - 1-Nitropyrene (1-NP) induces apoptosis and apparently a non-apoptotic programmed cell death (paraptosis) in Hepa1c1c7 cells. AB - Mechanistic studies of nitro-PAHs (polycyclic aromatic hydrocarbons) of interest might help elucidate which chemical characteristics are most important in eliciting toxic effects. 1-Nitropyrene (1-NP) is the predominant nitrated PAH emitted in diesel exhaust. 1-NP-exposed Hepa1c1c7 cells exhibited marked changes in cellular morphology, decreased proliferation and different forms of cell death. A dramatic increase in cytoplasmic vacuolization was observed already after 6 h of exposure and the cells started to round up at 12 h. The rate of cell proliferation was markedly reduced at 24 h and apoptotic as well as propidium iodide (PI)-positive cells appeared. Electron microscopic examination revealed that the vacuolization was partly due to mitochondria swelling. The caspase inhibitor Z-VAD-FMK inhibited only the apoptotic cell death and Nec-1 (an inhibitor of necroptosis) exhibited no inhibitory effects on either cell death or vacuolization. In contrast, cycloheximide markedly reduced both the number of apoptotic and PI-positive cells as well as the cytoplasmic vacuolization, suggesting that 1-NP induced paraptotic cell death. All the MAPKs; ERK1/2, p38 and JNK, appear to be involved in the death process since marked activation was observed upon 1-NP exposure, and their inhibitors partly reduced the induced cell death. The ERK1/2 inhibitor PD 98057 completely blocked the induced vacuolization, whereas the other MAPKs inhibitors only had minor effects on this process. These findings suggest that 1-NP may cause apoptosis and paraptosis. In contrast, the corresponding amine (1-aminopyrene) elicited only minor apoptotic and necrotic cell death, and cells with characteristics typical of paraptosis were absent. PMID- 18417181 TI - In vitro and in vivo evaluation of the effects of piperine on P-gp function and expression. AB - Piperine, a major component of black pepper, is used as spice and nutrient enhancer. The purpose of the present study was to evaluate the effects of acute and prolonged piperine exposure on cellular P-gp expression and function in vitro and in vivo. Piperine at concentrations ranging from 10 to 100 microM, determined by MTT assay to be non-cytotoxic, was observed to inhibit P-gp mediated efflux transport of [(3)H]-digoxin across L-MDR1 and Caco-2 cell monolayers. The acute inhibitory effect was dependent on piperine concentration, with abolishment of [(3)H]-digoxin polarized transport attained at 50 microM of piperine. In contrast, prolonged (48 and 72 h) co-incubation of Caco-2 cell monolayers with piperine (50 and 100 microM) increased P-gp activity through an up-regulation of cellular P-gp protein and MDR1 mRNA levels. The up-regulated protein was functionally active, as demonstrated by a higher degree of [(3)H]-digoxin efflux across the cell monolayers, but the induction was readily reversed by the removal of the spice from the culture medium. Peroral administration of piperine at the dose of 112 microg/kg body weight/day to male Wistar rats for 14 consecutive days also led to increased intestinal P-gp levels. However, there was a concomitant reduction in the rodent liver P-gp although the kidney P-gp level was unaffected. Our data suggest that caution should be exercised when piperine is to be co administered with drugs that are P-gp substrates, particularly for patients whose diet relies heavily on pepper. PMID- 18417180 TI - Activation of Nrf2 by arsenite and monomethylarsonous acid is independent of Keap1-C151: enhanced Keap1-Cul3 interaction. AB - Drinking water contaminated with arsenic, a human carcinogen, is a worldwide health issue. An understanding of cellular signaling events in response to arsenic exposure and rational designing of strategies to reduce arsenic damages by modulating signaling events are important to fight against arsenic-induced diseases. Previously, we reported that activation of the Nrf2-mediated cellular defense pathway confers protection against toxic effects induced by sodium arsenite [As(III)] or monomethylarsonous acid [MMA(III)]. Paradoxically, arsenic has been reported to induce the Nrf2-dependent signaling pathway. Here, we report the unique mechanism of Nrf2 induction by arsenic. Similar to tert butylhydroquinone (tBHQ) or sulforaphane (SF), arsenic induced the Nrf2-dependent response through enhancing Nrf2 protein levels by inhibiting Nrf2 ubiquitination and degradation. However, the detailed action of arsenic in Nrf2 induction is different from that of tBHQ or SF. Arsenic markedly enhanced the interaction between Keap1 and Cul3, subunits of the E3 ubiquitin ligase for Nrf2, which led to impaired dynamic assembly/disassembly of the E3 ubiquitin ligase and thus decreased its ligase activity. Furthermore, induction of Nrf2 by arsenic is independent of the previously identified C151 residue in Keap1 that is required for Nrf2 activation by tBHQ or SF. Distinct mechanisms of Nrf2 activation by seemingly harmful and beneficial reagents provide a molecular basis to design Nrf2-activating agents for therapeutic intervention. PMID- 18417182 TI - Gene expression changes induced by ochratoxin A in renal and hepatic tissues of male F344 rat after oral repeated administration. AB - Ochratoxin A (OTA), a naturally occurring mycotoxin, is nephrotoxic in all animal species tested and is considered a potent renal carcinogen, particularly in male rats. Its mechanism of toxicity is still unknown, although oxidative stress appears to be a plausible mechanism. Therefore, the objective of this study was to identify the biological pathways that are modulated in vivo by OTA in male F344 rats in order to gain further insight into its mechanism of renal toxicity. Rats were gavaged daily with OTA (500 microg/kg bw) and gene expression profiles in target and non-target organs were analyzed after 7 and 21 days administration. As was expected, a time-dependent increase of OTA concentrations was found in plasma, kidney and liver, with the concentrations found in both tissues being quite similar. However, histopathological examinations only revealed changes in kidney; signs of nephrotoxicity involving single cell necrosis and karyomegalic nuclei were observed in the treated rats. The number of differentially expressed genes in kidney was much higher than in liver (541 versus 11 at both time points). Several similarities were observed with other in vivo gene expression data. However, great differences were found with previous in vitro gene expression data, with the exception of DNA damage response which was not observed at mRNA level in any of our study conditions. Down-regulation was the predominant effect. Oxidative stress response pathway and genes involved in metabolism and transport were inhibited at both time points. RGN (regucalcin) - a gene implicated in calcium homeostasis - was strongly inhibited at both time points and genes implicated in cell survival and proliferation were up-regulated at day 21. Moreover, translation factors and annexin genes were up-regulated at both time points. Apart from oxidative stress, alterations of the calcium homeostasis and cytoskeleton structure may be present at the first events of OTA toxicity. PMID- 18417183 TI - Azaspiracid: first evidence of protein binding in shellfish. AB - The occurrence of azaspiracid (AZA) toxins in contaminated shellfish has been the focus of much research. The present study investigated the binding properties of these toxins in mussels of the species Mytilus edulis. The work involved extraction of proteins and AZAs from contaminated mussel hepatopancreas and examination of the extracts by isoelectric focusing (IEF), size exclusion chromatography (SEC) and sodium docecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). Liquid chromatography coupled with tandem mass spectrometry analysis (LC-MS/MS) was also performed in this study to identify AZAs. Blank mussels were subjected to the same purification and analytical procedures. AZAs were found to be weakly bound to a protein with a molecular weight of 45 kDa, in samples of contaminated mussels. This protein, which was abundant in contaminated mussels, was also present in blank mussels, albeit at much lower concentrations. It was further noted that a 22 kDa protein was also present only in contaminated mussel samples. PMID- 18417184 TI - The extraction of features and disparities from images by a model based on the neurological organisation of the visual system. AB - A computational simulation of the early stages of mammalian visual processing, from the retina to the primary visual cortex, is described. The simulation uses elements that are organised according to the anatomical connections of the biological visual system. It explores how observed responses of simple cells of the primary visual cortex can be generated by a small number of stages of the types of processing that are observed in the nervous system. Edge features are extracted from single images and disparities between stereoscopic image pairs are detected with good reliability. An important parameter affecting processing was found to be the strength of the surround inhibition between the elements that represent neurones of the primary visual cortex. PMID- 18417185 TI - Chromatic temporal integration and retinal eccentricity: psychophysics, neurometric analysis and cortical pooling. AB - Psychophysical chromatic sensitivity deteriorates in peripheral retina, even after appropriate size scaling of targets. This decrease is more marked for stimuli targeted at the long- (L) to middle-wavelength (M) cone opponent system than for stimuli targeted at short-wavelength (S) pathways. Foveal chromatic mechanisms integrate over several hundred milliseconds for pulse detection. If the time course for integration were shorter in the periphery, this might account for sensitivity loss. Psychophysical chromatic temporal integration (critical duration) for human observers was estimated as a function of eccentricity. Critical duration decreased by a factor of 2 (from approximately 200 to approximately 100 ms) from the fovea to 20 degrees eccentricity. This partly (but not completely) accounts for the decrease in /L-M/ sensitivity in the periphery, but almost completely accounts for the decrease in S-cone sensitivity. Some loss of /L-M/I sensitivity thus has a cortical locus. In a physiological analysis, we consider how the /L-M/ cone parvocellular pathway integrates chromatic signals. Neurometric contrast sensitivities of individual retinal ganglion cells decreased with the square-root of stimulus duration (as expected from Poisson statistics of ganglion cell firing). In contrast, psychophysical data followed an inverse linear relationship (Bloch's law). Models of cortical pooling mechanisms incorporating uncertainty as to stimulus onset and duration can at least partially account for this discrepancy. PMID- 18417186 TI - Plaid perception is only subtly impaired in strabismic amblyopia. AB - Amblyopes exhibit a global motion anomaly that implicates processing beyond the local motion analysis of V1 possibly involving areas MT and MST in the extra striate cortex. Here, we sought to further investigate this deficit by measuring the perception of moving plaid stimuli by amblyopic observers, since there is good physiological evidence that the motion of such stimuli is determined by processes beyond V1. The conditions under which the two moving components constituting the plaids were seen to cohere or move transparently over one another were investigated by manipulating their relative spatial frequencies. Percepts were measured using both short presentation durations, where both the percept and the direction of motion were reported, and long presentation durations where the bi-stability of the stimulus was directly measured. In addition, we measured the ability of amblyopic eyes to perceive globally coherent motion in a multiple aperture stimulus. We found a small increased tendency for both amblyopic and fellow-fixing eyes to perceive short duration plaid stimuli as coherent relative to control eyes, but no difference for long duration plaids. In addition, amblyopic eyes saw less coherence in multiple aperture stimuli than fellow-fixing eyes but were not reliably different from control eyes. We therefore conclude that the neural mechanisms underlying plaid perception are only subtly abnormal in amblyopia. PMID- 18417188 TI - Calculating human exposure to endocrine disrupting pesticides via agricultural and non-agricultural exposure routes. AB - Endocrine Disrupting Chemicals (EDCs) are of increasing concern because of their potential impacts on the environment, wildlife and human health. Pesticides and some pesticide metabolites are an important group of EDC, and exposure to them is a poorly quantified source of human and environmental exposure to such chemicals generally. Models for estimating human exposure to Endocrine Disrupting (ED) pesticides are an important risk management tool. Probabilistic models are now being used in addition to deterministic ones in all areas of risk assessment. These can provide more realistic exposure estimates, because they are better able to deal with variation and uncertainty more effectively and better inform risk management decisions. Deterministic models are still used and are of great value where exposure data are scarce. Models or groups of models that provide holistic human ED pesticide exposure estimates are required if the risk posed to humans by ED pesticides is to be better assessed. Much more research is needed to quantify different exposure routes such as exposure from agricultural spray drift and the medical use of pesticides to develop such models. Most available probabilistic models of human exposure were developed in the USA and require modification for use elsewhere. In particular, datasets equivalent to those used to create and apply the American models are required. This paper examines the known routes of human pesticide exposure with particular reference to ED pesticides and their quantification as unlike pesticides generally, many ED pesticides are harmful at very low doses, especially if exposure occurs during sensitive stages of development, producing effects that may not manifest for many years or that affect descendants via epigenetic changes. It also summarises available deterministic and probabilistic models commonly used to calculate human exposure. The main requirement if such models are to be used in the UK is more quantitative data on the sources and pathways of human ED pesticide exposure. PMID- 18417189 TI - Potential geographic distribution of atmospheric nitrogen deposition from intensive livestock production in North Carolina, USA. AB - To examine the consequences of increased spatial aggregation of livestock production facilities, we estimated the annual production of nitrogen in livestock waste in North Carolina, USA, and analyzed the potential distribution of atmospheric nitrogen deposition from confined animal feeding operations ("CAFO") lagoons. North Carolina is a national center for industrial livestock production. Livestock is increasingly being raised in CAFOs, where waste is frequently held, essentially untreated, in open-air lagoons. Reduced nitrogen in lagoons is volatilized as ammonia (NH(3)), transported atmospherically, and deposited to other ecosystems. The Albemarle-Pamlico Sound, NC, is representative of nitrogen-sensitive coastal waters, and is a major component of the second largest estuarine complex in the U.S. We used GIS to model the area of water in the Sound within deposition range of CAFOs. We also evaluated the number of lagoons within deposition range of each 1 km(2) grid cell of the state. We considered multiple scenarios of atmospheric transport by varying distance and directionality. Modeled nitrogen deposition rates were particularly elevated for the Coastal Plain. This pattern matches empirical data, suggesting that observed regional patterns of reduced nitrogen deposition can be largely explained by two factors: limited atmospheric transport distance, and spatial aggregation of CAFOs. Under our medium-distance scenario, a small portion (roughly 22%) of livestock production facilities contributes disproportionately to atmospheric deposition of nitrogen to the Albemarle-Pamlico Sound. Furthermore, we estimated that between 14-37% of the state receives 50% of the state's atmospheric nitrogen deposition from CAFO lagoons. The estimated total emission from livestock is 134,000 t NH(3) yr(-1), 73% of which originates from the Coastal Plain. Stronger waste management and emission standards for CAFOs, particularly those on the Coastal Plain nearest to sensitive water bodies, may help mitigate negative impacts on aquatic ecosystems. PMID- 18417187 TI - Mitochondrial DNA repair in aging and disease. AB - Mitochondria are organelles which, according to the endosymbiosis theory, evolved from purpurbacteria approximately 1.5 billion years ago. One of the unique features of mitochondria is that they have their own genome. Mitochondria replicate and transcribe their DNA semiautonomously. Like nuclear DNA, mitochondrial DNA (mtDNA) is constantly exposed to DNA damaging agents. Regarding the repair of mtDNA, the prevailing concept for many years was that mtDNA molecules suffering an excess of damage would simply be degraded to be replaced by newly generated successors copied from undamaged genomes. However, evidence now clearly shows that mitochondria contain the machinery to repair the damage to their genomes caused by certain endogenous or exogenous damaging agents. The link between mtDNA damage and repair to aging, neurodegeneration, and carcinogenesis associated processes is the subject of this review. PMID- 18417190 TI - Evolution of the source apportionment of the lipidic fraction from sediments along the Fensch River, France: a multimolecular approach. AB - The Fensch River (FR) is one of the most contaminated rivers in France due to the population density and the concentration of industrial activities in this small watershed area. From upstream to downstream, the organic matter extracted from sediments has been analyzed by gas chromatography-mass spectrometry and molecules have been quantified and classified into natural, petrogenic, pyrogenic and sewage water (SW) markers. Upstream the river, anthropogenic molecules are already predominant and represent 87.1% of the molecules quantified. This proportion increases from upstream to downstream and rises to 96.8% at the confluence of the FR with the Moselle River. In the upper part of the FR the contamination is mainly due to human waste (coprostanol: 36.44 microg/g; 42.1% of anthropogenic markers). In the lower part, the contribution of SW markers decreases from 42.1 to 2.4% and the proportion of pyrogenic molecules increases from 29.6 to 59.6%. The major sources of pyrogenic organic matter have been determined by calculation of specific ratios on polycyclic aromatic hydrocarbons and by comparison with reported data. Coal tar, road runoff and atmospheric depositions of urban particles seem to be the major pyrogenic sources. Along the river, the proportion of petrogenic molecules remains constant and those molecules seem to be mainly inherited from road runoff, in the upper part of the FR. Industrial lubricants that occur in steel plant sludge are an additional source in the lower part of the river. PMID- 18417191 TI - The diffuse infiltration of road runoff: an environmental improvement. AB - The Laboratory of Engineering and Environmental Geology (GEOLEP) has been mandated by Swiss authorities (Swiss Federal Road Office FedRO) to test a new road runoff management concept. This concept promotes the diffuse infiltration of road runoff into infiltration slopes designed for this purpose. Soils retain particles and contaminants; this lowers the road impact on the environment and simultaneously improves aquifer recharge. This concept has to be adapted to aquifer vulnerability and traffic conditions. Thus, a real-scale experimental station was designed and built in Switzerland to assess the feasibility of this new concept. This station allowed the testing of two lysimeters composed of 80 cm of A and B-horizons. Water and chemical fluxes were measured at the lysimeter bases. Road runoff was sampled in a distinct collector. Infiltration of road runoff into the local aquifer was monitored thanks to six piezometers. Water quality and quantity were therefore measured at each step of the infiltration process. Results provided by 112 natural events showed that soil horizons accommodated road runoff flows. The least favourable conditions for contaminant retention are encountered during thunderstorms, when high concentrations of substances deposited on the road are remobilised within a short time and rapidly percolates through soil horizons. Thus, three artificial events were designed to mimic heavy thunderstorms. Concentrations measured in road runoff notably decreased after soil filtration. Substances with high distribution coefficients Kd (low mobility) had concentrations reduced to lower values (1/1000 to 1/10,000), while those with high mobility retained similar concentrations even after soil filtration. However, these mobile substances exhibited low concentrations in the underlying aquifer due to dilution. This innovative road runoff management concept can thus be readily implemented outside groundwater protection zones where aquifers are slightly vulnerable; it undoubtedly lowers the environmental impact of roads, does not endanger road integrity, and locally increases aquifer recharge. PMID- 18417192 TI - Influence of smoking during pregnancy on haemostasis in healthy full term neonates. AB - BACKGROUND: Clinical and experimental researches have linked smoking to disturbances of coagulation and fibrinolysis. Several potential mechanisms are incriminated involving inflammation, fibrinogen synthesis and clotting factors. Based on the fact that the majority of tobacco components cross the placental barrier, the objective of our current study is to investigate the influence of smoking during pregnancy on neonatal haemostasis. STUDY DESIGN: The study was based on a comparative evaluation of coagulation and fibronolysis between healthy full term infants of women who smoked during pregnancy and a control group. Subjects consisted of 39 newborns of smoking and 43 newborns of nonsmoking mothers. Blood samples were obtained shortly after birth and before the administration of vitamin K. Investigation included: PT, INR, aPTT, fibrinogen, coagulation factors II, V, VII, VIII, IX, X, XI, XII, vWillebrand (vWF), protein C and S, APCr, anti-thrombin (AT), t-PA and PAI-1. The independent t- test was used to compare the differences between the values of coagulation and fibrinolytic parameters at the p<0.05 level. RESULTS: We discovered a statistically significant decrease in factor II and protein S levels and an elevation in t-PA and factor VIII concentrations in newborns of smoking mothers, without clinical manifestations of altered haemostasis. There were no significant differentiations in other coagulation or fibrinolytic parameters. CONCLUSION: The alteration in factor II, protein S, t-PA and factor VIII in neonates exposed in utero to tobacco smoke is not accompanied by loss in the balance between coagulation and fibrinolytic pathways. PMID- 18417193 TI - Enhancing effect of the (145)Met-allele of GPIb alpha on platelet sensitivity to aspirin under high-shear conditions. PMID- 18417194 TI - Endothelial haemostatic markers in members of families with familial combined hyperlipidemia. AB - INTRODUCTION: The aim of this study was to evaluate the plasma levels of endothelial haemostatic markers - von Willebrand factor (vWF), plasminogen activator inhibitor-1 (PAI-1), tissue plasminogen activator (t-PA) and soluble thrombomodulin (sTM) - in asymptomatic, nonsmoking members of families with familial combined hyperlipidemia (FCH). We investigated the association between these factors and the intima-media thickness (IMT) of the common carotid artery, selected risk factors of atherosclerosis and markers of insulin resistance. METHODS: 82 members of 29 FCH families were divided into two groups: HL (probands and hyperlipidemic first-degree relatives, n=47) and NL (normolipidemic first degree relatives, n=35). The control groups C-HL (n=20) and C-NL (n=20) consisted of sex- and age-matched healthy individuals. IMT was measured by ultrasound at a far wall of both common carotid arteries. RESULTS: Compared with healthy controls, hyperlipidemic subjects had significantly higher levels of vWF (146.4+/ 73.2% versus 112.2+/-29.4%, p<0.05), of PAI-1 (102.4[83.0-117.0] ng/ml versus 63.5[31.8-87.3] ng/ml, p<0.01) and of t-PA (5.1[2.5-7.9] ng/ml versus 3.4[1.4 5.8] ng/ml, p<0.05). They had increased IMT, which correlated with vWF (r=0.29, p<0.05). Their normolipidemic relatives had significantly higher levels of vWF (137.2+/-42.8% versus 106.6+/-24.0%, p<0.01) and of PAI-1 (75.3[53.2-92.0] ng/ml versus 48.6[37.4-85.9] ng/ml, p<0.05). Levels of vWF, PAI-l and t-PA were independently associated with several markers of insulin resistance. CONCLUSIONS: Asymptomatic members of FCH families have increased endothelial haemostatic factors- vWF, PAI-1, t-PA, which are associated with insulin resistance. VWF correlates with morphological vascular changes, detected by the increase of IMT, presented in only hyperlipidemic subjects. PMID- 18417195 TI - Variations in platelet protein associated with arterial thrombosis. AB - INTRODUCTION: Hyperactivity of platelets has been associated with thrombotic episodes by molecular mechanisms not yet elucidated. The present work aimed at identifying whether the platelet protein content from patients who had suffered an arterial thrombosis episode differed from that of platelets obtained from normal healthy donors. METHODS: Differential platelet protein profiles were determined by 2-dimensional (2-D) gel electrophoresis and Western blot analysis of total platelet lysates. Identification of differentially expressed proteins was carried out by mass spectrometry (MALDI-TOF). RESULTS: We found a decreased platelet content of three protein spots in patients of arterial thrombosis: integrin linked kinase (ILK), fructose bisphosphate aldolase (aldolase) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) whereas the content of four other protein spots was increased: actin binding protein, coronine like (p57), non-muscle myosin heavy chain (NMMHC-A), pyruvate kinase M2 isoenzyme (PK) and phosphoglycerate kinase (PGK). The variations in ILK, GAPDH and PK were validated by Western blot analysis. The proteins showing a decreased platelet content in arterial thrombosis patients are associated with the cytoskeletal insoluble fraction and the detected increase in some proteins seems to be due to the generation of peptides caused by a limited proteolysis. Differences in the protein profiles of circulating platelets from arterial thrombosis were maintained months after the acute thrombotic event and disappear in the long term. CONCLUSIONS: The observed variations in some platelet proteins suggest the existence of a perturbation in the cytoskeletal organization and increased proteolysis, both indicative of a platelet pro-active state, persistent after the thrombotic event. PMID- 18417196 TI - Cardiac resynchronization: insight from experimental and computational models. AB - Cardiac resynchronization therapy (CRT) is a promising therapy for heart failure patients with a conduction disturbance, such as left bundle branch block. The aim of CRT is to resynchronize contraction between and within ventricles. However, about 30% of patients do not respond to this therapy. Therefore, a better understanding is needed for the relation between electrical and mechanical activation. In this paper, we focus on to what extent animal experiments and mathematical models can help in order to understand the pathophysiology of asynchrony to further improve CRT. PMID- 18417199 TI - Association of built-environment, social-environment and personal factors with bicycling as a mode of transportation among Austrian city dwellers. AB - OBJECTIVE: To determine the association of built-environment, social-environment, and personal-level factors with bicycling for transportation, among adult city dwellers. METHOD: Survey of a representative sample of 1000 inhabitants of the city of Graz, Austria, using a computer-assisted telephone interview addressing cycling behavior and associated personal, social and environmental factors. RESULTS: The prevalence of biking for transportation was 22.5%. After adjustment for gender, age, education, physical activity level and distance from home to destination, cycling was positively associated with the presence of bike lane connectivity (OR=2.09) and social support/modeling (OR=1.62), and negatively associated with the perceived barriers of "physical discomfort" (OR=0.49) and "an impractical transport mode" (OR=0.50). Analysis of interactions indicated that the effect of the perceived benefit of "rapidity" was stronger in physically active persons than inactive individuals, and the effect of the perceived barrier of "an impractical mode of transportation" was stronger among women than men. CONCLUSION: In addition to cycling-related social support and perceived benefits and barriers, bike lane connectivity may be an important determinant of cycling as a means of transportation among adult city dwellers. PMID- 18417198 TI - Alveolar macrophage phagocytosis is impaired in children with poorly controlled asthma. AB - BACKGROUND: Lower respiratory tract infection is a differentiating feature of children with poorly controlled asthma. OBJECTIVE: Given the role of alveolar macrophages (AMs) in innate immunity, we hypothesized that AM phagocytosis might be impaired in poorly controlled asthma. METHODS: Bronchoalveolar lavage fluid AMs were isolated from 28 asthmatic children (moderate asthma, n = 12; severe asthma, n = 16), 10 nonasthmatic children with chronic cough treated with inhaled corticosteroids, and 10 healthy adult control subjects. AMs were stimulated with LPS and exposed to fluorescein isothiocyanate-conjugated Staphylococcus aureus for 2 hours. Phagocytosis was quantified by using a phagocytic index (PI) calculated from the percentage of phagocytic cells multiplied by the relative fluorescence (RFU) units of S. aureus per cell. Apoptosis was determined from the percentage of cells positive for poly (adenosine diphosphate-ribose) polymerase. RESULTS: Phagocytosis as measured by using the unstimulated PI was decreased in subjects with poorly controlled asthma (healthy control subjects, 9330 +/- 3992 RFU; chronic cough, 9042 +/- 5976 RFU; moderate asthma, 4361 +/- 2536 RFU; severe asthma, 3153 +/- 1886 RFU; P < .001) and remained unchanged with LPS stimulation. Children with severe asthma also had increased AM apoptosis, both the unstimulated and LPS-simulated states (P < .001), which correlated with the PI. CONCLUSIONS: AM function is compromised in children with poorly controlled asthma and is characterized by decreased phagocytosis and increased apoptosis. PMID- 18417200 TI - Motor programming in apraxia of speech. AB - Apraxia of Speech (AOS) is an impairment of motor programming. However, the exact nature of this deficit remains unclear. The present study examined motor programming in AOS in the context of a recent two-stage model [Klapp, S. T. (1995). Motor response programming during simple and choice reaction time: The role of practice. Journal of Experimental Psychology: Human Perception and Performance, 21, 1015-1027; Klapp, S. T. (2003). Reaction time analysis of two types of motor preparation for speech articulation: Action as a sequence of chunks. Journal of Motor Behavior, 35, 135-150] that proposes a preprogramming stage (INT) and a process that assigns serial order to multiple programs in a sequence (SEQ). The main hypothesis was that AOS involves a process-specific deficit in the INT (preprogramming) stage of processing, rather than in the on line serial ordering (SEQ) and initiation of movement. In addition, we tested the hypothesis that AOS involves a central (i.e., modality-general) motor programming deficit. We used a reaction time paradigm that provides two dependent measures: study time (the amount of time for participants to ready a motor response; INT), and reaction time (time to initiate movement; SEQ). Two experiments were conducted to examine INT and SEQ in AOS: Experiment 1 involved finger movements, Experiment 2 involved speech movements analogous to the finger movements. Results showed longer preprogramming time for patients with AOS but normal sequencing and initiation times, relative to controls. Together, the findings are consistent with the hypothesis of a process-specific, but central (modality-independent) deficit in AOS; alternative explanations are also discussed. PMID- 18417201 TI - Masked repetition priming using magnetoencephalography. AB - Masked priming is used in psycholinguistic studies to assess questions about lexical access and representation. We present two masked priming experiments using MEG. If the MEG signal elicited by words reflects specific aspects of lexical retrieval, then one expects to identify specific neural correlates of retrieval that are sensitive to priming. To date, the electrophysiological evidence has been equivocal. We report findings from two experiments. Both employed identity priming, where the prime and target are the same lexical item but differ in case (NEWS-news). The first experiment used only forward masking, while the prime in the second experiment was both preceded and followed by a mask (backward masking). In both studies, we find a significant behavioral effect of priming. Using MEG, we identified a component peaking approximately 225 ms post onset of the target, whose latency was sensitive to repetition. These findings support the notion that properties of the MEG response index specific lexical processes and demonstrate that masked priming can be effectively combined with MEG to investigate the nature of lexical processing. PMID- 18417202 TI - Comparison of the bond strength of laser-sintered and cast base metal dental alloys to porcelain. AB - OBJECTIVE: The purpose of this study was to compare shear bond strengths of cast Ni-Cr and Co-Cr alloys and the laser-sintered Co-Cr alloy to dental porcelain. METHODS: Dental porcelain was applied on two cast and one laser-sintered base metal alloy. Ten specimens were prepared for each group for bond strength comparison. ANOVA followed by Tukey HSD multiple comparison test (alpha=0.05) was used for statistical analysis. Fractured specimens were observed with a stereomicroscope to classify the type of failure after shear bond testing. RESULTS: While the mean shear bond strength was highest for the cast Ni-Cr metal ceramic specimens (81.6+/-14.6 MPa), the bond strength was not significantly different (P>0.05) from that for the cast Co-Cr metal-ceramic specimens (72.9+/ 14.3 MPa) and the laser-sintered Co-Cr metal-ceramic specimens (67.0+/-14.9 MPa). All metal-ceramic specimens prepared from cast Ni-Cr and Co-Cr alloys exhibit a mixed mode of cohesive and adhesive failure, whereas five of the metal-ceramic specimens prepared from the laser-sintered Co-Cr alloy exhibited the mixed failure mode and five specimens exhibited adhesive failure in the porcelain. CONCLUSIONS: The new laser-sintering technique for Co-Cr alloy appears promising for dental applications, but additional studies of properties of the laser sintered alloy and fit of castings prepared by this new technique are needed before its acceptance into dental laboratory practice. SIGNIFICANCE: Laser sintering of Co-Cr alloy seems to be an alternative technique to conventional casting of dental alloys for porcelain fused to metal restorations. PMID- 18417203 TI - Suitability evaluation of sol-gel derived Si-substituted hydroxyapatite for dental and maxillofacial applications through in vitro osteoblasts response. AB - Si-hydroxyapatite (Si-HAP) has been used in orthopedic, dental, and maxillofacial surgery as a bone substitute. OBJECTIVE: The aim of this investigation was to study the effect of Si substitution into the hydroxyapatite matrices and evaluate the biocompatibility effects of Si-HAP material in vitro with human osteoblasts. METHODS: Silicon-substituted hydroxyapatite (Si-HAP) bioceramic materials were prepared by incorporating small amounts of silicon into the structure of hydroxyapatite [Ca10(PO4)6(OH)2, HAP] through a sol-gel method. A series of silicon substitutions ranging from 0, 1, 3 and 5 mol%, which are comparable to the measured silicon contents in natural bone, were performed. RESULTS: Single phase Si-HAP was obtained upon calcining the as-prepared powders up to 800 degrees C since no secondary phases, such as tricalcium phosphate (TCP), tetracalcium phosphate (TeCP) or calcium oxide (CaO), were identified by X-ray diffraction analysis. The effects of silicon-substituted hydroxyapatite (Si-HAP) materials towards the responses of human osteoblast-like (HOB) cells were investigated and compared with pure hydroxyapatite. SIGNIFICANCE: The Si-HAP indicated a significant increase in cell growth density with culture time irrespective of the amount of Si substituted in HAP. A high Si content (5 mol%) appears to promote rapid bone mineralization, since large amount of calcium phosphate minerals started to develop across the ECM by day 31 for a sample containing 5 mol% Si. On the other hand, a high Si content may result in fast dissolution of the material, owing to a decrease of HAP crystallite size, which might not be ideal for cell attachment for prolonged time periods. An optimum level of Si appears to exist at 3 mol%, which balances these effects. PMID- 18417204 TI - Innovations in bonding to zirconia-based materials: Part I. AB - OBJECTIVES: Establishing a reliable bond to zirconia-based materials has proven to be difficult which is the major limitation against fabricating adhesive zirconia restorations. This bond could be improved using novel selective infiltration etching conditioning in combination with engineered zirconia primers. Aim of the work was to evaluate resin-to-zirconia bond strength using selective infiltration etching and novel silane-based zirconia primers. METHODS: Zirconia discs (Procera Zirconia) received selective infiltration etching surface treatment followed by coating with either of five especially engineered experimental zirconia primers. Pre-aged resin-composite discs (Tetric Ivo Ceram) were bonded to the treated surface using an MDP-containing resin-composite (Panavia F 2.0). The bilayered specimens were cut into microbars and the microtensile bond strength (MTBS) was evaluated. 'As-sintered' zirconia discs served as a control (alpha=0.05). The broken microbars were examined using a scanning electron microscope (SEM). RESULTS: The combination of selective infiltration etching with experimental zirconia primers significantly improved (F=3805, P<0.0001) the MTBS values (41+/-5.8 MPa) compared to the 'as-sintered' surface using the same primers which demonstrated spontaneous failure and very low bond strength values (2.6+/-3.1 MPa). SEM analysis revealed that selective infiltration etching surface treatment resulted in a nano-retentive surface where the zirconia primers were able to penetrate and interlock which explained the higher MTBS values observed for the treated specimens. PMID- 18417205 TI - Gene expression profiles of fathead minnows exposed to surface waters above and below a sewage treatment plant in Minnesota. AB - Water treatment plants are often not effective in removing pharmaceuticals, personal care products or natural hormones from their effluents. To test the effects of these effluents on fish, we exposed male fathead minnows (Pimephales promelas) for 48h to effluents at two sites, one up-stream and the other down stream from a water treatment plant. Gene expression profiling showed that significant changes occurred in the gonad of fish exposed below, compared to above the treatment plant and to laboratory control fish. Among the biological processes affected were the innate immune response, response to stress, control of homeostasis, control of transcription, metabolism, and cell communication. This work suggests that fish are impacted by exposures to the sewage treatment effluents and effects can be detected rapidly by gene expression profiling. PMID- 18417207 TI - Sutureless amniotic membrane transplantation for ocular surface reconstruction with a chemically defined bioadhesive. AB - The purpose of this study was to evaluate the efficiency and safety of a sutureless amniotic membrane transplantation (AMT) for ocular surface reconstruction with a chemically defined bioadhesive (CDB). The CDB was synthesized from aldehyded polysaccharides and epsilon-poly(L-lysine), two kinds of medical and food additives, as starting materials. Biocompatibility assay indicated that the CDB showed excellent biocompatibility with in vitro and in vivo ocular surface tissues and most of the CDB was histologically degraded within 4 weeks. Sutureless AMT using the CDB was safely and successfully performed onto a rabbit scleral surface. Transplanted amniotic membrane (AM) evaluated by histological, electron microscopic- and immunohistochemical examination indicated that the CDB did not affect normal differentiation of the cells or the integrity of the surrounding tissue. Thus, this newly developed CDB was found to be very useful for sutureless AMT for ocular surface reconstruction, without considering the risk of infection. It has the ability to fix AM to the ocular surface for a long time-period without additional inflammation, scarring, or damage to the surrounding tissues. PMID- 18417206 TI - Modification of silk fibroin using diazonium coupling chemistry and the effects on hMSC proliferation and differentiation. AB - A simple chemical modification method using diazonium coupling chemistry was developed to tailor the structure and hydrophilicity of silk fibroin protein. The extent of modification using several aniline derivatives was characterized using UV-vis and 1H NMR spectroscopies, and the resulting protein structure was analyzed with ATR-FTIR spectroscopy. Introduction of hydrophobic functional groups facilitated rapid conversion of the protein from a random coil to a beta sheet structure, while addition of hydrophilic groups inhibited this process. hMSCs were grown on these modified silks to assess the biocompatibility of these materials. The hydrophilicity of the silk derivatives was found to affect the growth rate and morphology, but hMSCs were able to attach, proliferate and differentiate into an osteogenic lineage on all of the silk derivatives. PMID- 18417208 TI - Reactive oxygen species inhibit polycystin-2 (TRPP2) cation channel activity in term human syncytiotrophoblast. AB - Pregnancy is often associated with oxidative stress (OS) and lower antioxidant defences, which are both implicated in the pathophysiology of preeclampsia, free radical-induced birth defects, and abortions, as well as gestational diabetes mellitus (GDM), where products of lipid peroxidation are increased. The molecular target(s) of increased oxygen free radicals and consequent lipid peroxidation in the human placenta remains ill defined. The human syncytiotrophoblast (hST) expresses abundant polycystin-2 (PC2, TRPP2), a TRP-type Ca(2+)-permeable non selective cation channel. Here, we explored the effect of reactive oxygen species (ROS) on PC2 channel activity of term hST. Apical membranes of the hST were reconstituted in a lipid bilayer chamber. Addition of either hydrogen-peroxide (H(2)O(2)) or tert-butyl hydroperoxide (tBHP) to the cis chamber (intracellular side) rapidly and completely inhibited PC2-mediated cation channel activity in reconstituted hST vesicles. A dose-response titration with increasing concentrations of H(2)O(2) gave an IC(50)=131 nM. The effect of H(2)O(2) on the isolated protein from in vitro transcribed/translated material was significantly different. H(2)O(2) inhibited PC2 cation channel activity, with a much lower affinity (IC(50)=193 microM). To correlate these findings with H(2)O(2)-induced lipid peroxidation, TBARS where measured in hST apical membranes incubated with H(2)O(2). Increased TBARS by exposure of hST apical membranes to H(2)O(2) (625 microM) returned to control value in the presence of catalase (167 microg/ml). Taken together these data indicate that ROS affect PC2 channel function by targetting both membrane lipids and the channel protein. Thus, OS in human pregnancy may be linked to dysregulation of channels such as PC2, which allow the transport of Ca(2+) into the placenta. Oxidative complications in pregnancy may implicate dysfunctional cation transfer between mother and fetus. PMID- 18417209 TI - Contractility of placental vascular smooth muscle cells in response to stimuli produced by the placenta: roles of ACE vs. non-ACE and AT1 vs. AT2 in placental vessel cells. AB - Our previously published work has shown that non-ACE angiotensin II (Ang II) generating system is dominate in the placenta and may play a critical role in regulation of placental vascular contractile function. In the present study, using a collagen gel contraction assay we further studied contractility of placental vascular smooth muscle cells (VSMCs) in response to factors produced by preeclamptic (PE) placentas. Placental VSMCs/type-1 collagen gels were incubated with PE placental conditioned medium in the presence or absence of inhibitors or receptor blockers. Captopril (an ACE inhibitor), chymostatin (a non-ACE chymase inhibitor), losartan (an AT1 receptor blocker) and PD123,319 (an AT2 receptor blocker) were used to study the specific ACE vs. non-ACE and AT1 vs. AT2 effects on placental VSMC contractility, respectively. Our results showed that chymostatin, but not captopril, and PD123,319, but not losartan, significantly attenuated placental VSMC/collagen gel contraction, p<0.01, respectively. The inhibitory effects of chymostatin and PD123,319 were dose-dependent. Our results suggest that chymase, a non-ACE Ang II generating enzyme, may contribute significantly to Ang II generated in the placenta vascular tissue and that the AT2 receptor may play an important role in the regulation of Ang II induced contractility of placental VSMCs. These results provide new insights into Ang II generation and Ang II receptor regulation of vessel contractile function in the placental vasculature. These results also suggest the potential role of increased chymase activity and altered AT2 receptor function in placental related pregnancy disorders such as preeclampsia and IUGR. PMID- 18417210 TI - Effect of variable long-term maternal feed allowance on the development of the ovine placenta and fetus. AB - Maternal feed allowance during pregnancy can affect the development of the ovine placenta and fetus. The impact of variations in feed allowance prior to as well as throughout pregnancy has received less attention. Ewes were offered 0.6 (R), 1.2 (C) or 1.8 (AL) maintenance requirements from 89 days before conception until day 133 of pregnancy. Ewes were euthanised on days 50, 92 and 133 of pregnancy. Ewe live weight and body condition score, maternal and fetal metabolic and hormonal profiles, fetal body dimensions and organ weights, and the number, weight and morphology of placentomes were measured. Maternal live weight and condition score were lower in R compared to AL ewes at all stages of pregnancy (P<0.05). Plasma glucose and albumin concentrations of R ewes were significantly reduced (P<0.05) at mid and late gestation, respectively. Placental components were generally unresponsive to long term variations in maternal feed allowance. However, placental weight was significantly (P<0.05) correlated with fetal weight at days 50 (r=0.59) and 133 (r=0.69) of gestation. By late gestation growth retarded singleton fetuses from R ewes were 19% lighter (P<0.05), with reduced abdominal (9%) and thoracic (10%) girths (P<0.05) but of similar crown-rump length compared with fetuses from AL ewes. These differences were associated with significantly reduced IGF-I concentrations in fetal plasma (P<0.05). In conclusion, maternal, placental and fetal adaptations to long established planes of variable maternal feed allowance were able to maintain fetal growth during early and mid-pregnancy while fetal growth restriction, associated with reduced fetal IGF-I levels, became apparent in late pregnancy. PMID- 18417211 TI - Calcium signalling in wound-responsive normal human urothelial cell monolayers. AB - Epithelial tissue repair requires coordination of migratory and proliferative activity both adjacent to and remote from the wound edge. Although calcium signalling is implicated, the specific mechanisms are poorly understood. This study characterises the calcium signal invoked in response to scratch wounding of normal human urothelial (NHU) cells and relates it to the localised cellular response. Immediately after wounding of confluent NHU cell monolayers, cells adjacent to the wound edge showed a sustained (>30 min) rise in [Ca(2+)](i), while there was an independent, but simultaneous calcium wave that propagated out from the wound edge. The transient signal involved release of calcium from intracellular stores and was not mediated via gap junctions, but by diffusion of extracellular agonists. We demonstrated that ATP was partially responsible for the initiation and propagation of the calcium wave and showed that the calcium release mechanism was mediated in part via activation of inositol-1,4,5 triphosphate (IP(3)) receptors. By contrast, the sustained calcium signal originated from the extracellular milieu and correlated with an increased rate of migration by these cells. The work presented here provides supportive evidence that the calcium signature, defined by its temporal and amplitude characteristics, is important in co-ordinating the response of cells within an epithelial cell monolayer after wounding. PMID- 18417212 TI - Low erythropoietin production as non-oncogenic co-factor contributing to disease manifestation in low-risk MDS: a hypothesis supported by unique case reports. AB - Little is known about factors contributing to disease-evolution in early-phase myelodysplastic syndromes (MDS). In this article, low erythropoietin (EPO) production is discussed as a 'non-oncogenic' co-factor responsible for disease manifestation in a group of patients with low-risk MDS. The hypothesis is based on the observations that (i) individuals with bone marrow dysplasia and MDS related karyotypes but relatively high EPO levels may present without anemia and thus without frank MDS over years, (ii) several elderly patients with idiopathic anemia are low EPO producers and their anemia can be corrected with EPO therapy, and (iii) the well-known fact that low-risk MDS patients typically respond to EPO therapy when they have low endogenous EPO levels. PMID- 18417213 TI - Megakaryocytic blast crisis as a presenting manifestation of chronic myeloid leukemia. AB - Acute megakaryocytic leukemia is a rare form of acute myelogenous leukemia and may occur either de novo or by transformation of a preexisting myelodysplastic or myeloproliferative process including blast crisis of chronic myeloid leukemia (CML). Megakaryocytic blast crisis as the presenting manifestation of CML is extremely rare. We describe such a patient with no prior hematologic disease who presented with acute megakaryoblastic leukemia and extramedullary involvement, in whom the leukemic cells carried the BCR-ABL1 translocation as part of a complex karyotype. Using targeted sequential fluorescence in situ hybridization (T-FISH) technique, we detected two copies of BCR-ABL1 fusion gene in the leukemic blasts while the neutrophils carried a single copy of BCR-ABL1 fusion gene, thereby proving the origin of the megakaryoblastic leukemia from a previously undiagnosed CML clone. Blast crisis as a presenting manifestation of CML is rare and detecting clonal evolution of acute leukemia by specialized cytogenetic techniques may have important diagnostic and therapeutic implications. PMID- 18417214 TI - Improvement of cardiac function, mitral regurgitation and pulmonary hypertension in a patient with chronic eosinophilic leukemia (CEL) after low dose imatinib therapy. PMID- 18417216 TI - Ecotoxicological assessment of water quality and ecosystem health: a case study of the Volga River. AB - A comprehensive assessment is presented of the ecotoxicological situation in the Volga River basin from the viewpoint of ecosystem health. Concentrations of organic and inorganic toxic substances in water are reported. Basic clinical and postmortem signs of fish intoxication are described; changes in the cellular structure of their organs and tissues, as well as disturbances in hemogenesis, developing under the effect of toxic agents, are characterized. The comparative characteristics of several microelements (Ni, Cu, Sr, Al, Zn, Co, Mn, Pb, Cd, Hg, As) found in fish are presented. The main disturbances to fish caused by the accumulation of microelements in their organs and tissues are also considered. Based on dose-effect dependencies calculated with respect to the total concentration of toxic substances, standardized to MPC, and fish health criteria, cases that exceed the critical levels of pollutants are demonstrated for the investigated river sections. PMID- 18417215 TI - Fibrinogen-bearing protein genes in the snail Biomphalaria glabrata: characterization of two novel genes and expression studies during ontogenesis and trematode infection. AB - All fibrinogen (FBG)-bearing proteins documented to date in the freshwater snail Biomphalaria glabrata, the intermediate host of the human blood fluke Schistosoma mansoni, possess the same molecular structure; one or two immunoglobin superfamily (IgSF) domains at the N-terminus and a FBG domain at the C-terminus (named as FBG-related protein (FREP)). Here we report two novel genes that encode FBG-bearing proteins from B. glabrata. Different from all known FREPs, the first gene encodes a protein (657 amino acids (aa)) composed of a long N-terminal region with no sequence homology to any known protein, a middle epidermal growth factor (EGF) repeat region and a C-terminal FBG domain, designated FBG-related molecule (FReM). Differential expression at 2 days post-exposure (dpe) to the trematode S. mansoni or Echinostoma paraensei was found in the S. mansoni susceptible M line and resistant BS-90 snail strains. The second gene is a new member of the FREP family, designated FREP14, which encodes a 399 aa putative secreted protein. FREP14 is different from known FREPs in that it is encoded by a single locus and is not upregulated in early or late stage S. mansoni exposure, but is upregulated in late stage E. paraensei infection. Furthermore, gene expression during the snail's ontogenesis and at a late stage of trematode infection (52 dpe) has been investigated in the two newly identified genes (FReM and FREP14) described in this paper and five representative members of known FREPs (FREPs 2, 3, 4, 12, and 13). A variety of expression patterns were observed, suggestive of functional diversity among the members of FBG-bearing proteins. Our findings further broaden our understanding of the diversity and function of the FBG-bearing protein encoded genes in B. glabrata. PMID- 18417217 TI - The effects of treated effluents on the intensity of papillomatosis and HSP70 expression in roach. AB - Epidermal papillomatosis in fish has been proposed as an indicator of environmental stress but experimental evidence of connection between contaminants and papillomatosis in fish is scarce. We studied changes in the intensity of epidermal papillomatosis and the expression of heat shock protein 70 (HSP70) in roach, Rutilus rutilus, exposed to treated pulp mill and municipal effluents. In male roach, the increase in papillomatosis intensity was higher in fish exposed to 15% than in fish exposed to 1.5% concentration of municipal effluent. No differences were observed in papillomatosis development in females, or in HSP70 expression. In all the experiments conducted, the increasing effect of effluents seemed to be more pronounced in male fish suggesting that sex-related factors affected the intensity of papillomatosis after exposure to effluents. The present results indicate that environmentally relevant concentrations of municipal effluents may be contributing to the development of papillomatosis in fish. PMID- 18417218 TI - Changes in blood pesticide levels in booted eagle (Hieraaetus pennatus) associated with agricultural land practices. AB - We estimated the degree of exposure to lindane and endosulfan in the blood of booted eagle nestlings (Hieraaetus pennatus) (1999-2003), in order to assess the usefulness of these samples as a unit for monitoring changes in exposure as a result of shifts in agricultural practices and the implementation of legal measures. The highest blood lindane concentrations were obtained 1 year prior to its prohibition by the European Union. Subsequent to that year, the drop in blood concentrations was dramatic. Furthermore, endosulfan blood concentrations follow a progression coinciding with an increase in olive, grape and plum-tree crops. We conclude that concentrations of organochlorine pesticides in the blood of booted eagle nestlings may be used to monitor the use of those pesticides over a particular agricultural region and alert the authorities of possible environmental or health risks. PMID- 18417219 TI - [Unruptured intracranial aneurysm and microsurgical exclusion: the need of a randomized study of surgery versus natural history]. AB - Although unruptured intracranial aneurysm (UIA) is becoming a more common finding nowadays, determining the optimal treatment strategy is difficult because the risk of rupture is poorly understood and surgery is not without its own hazards. As the mortality rate after rupture is estimated to range from 56 to 83%, the final therapeutic decision is the result of an analysis of rupture risk and the risks related to surgical exclusion, which may be determined by consideration of the risk factors. We considered the UIA to have a high risk of rupture if it was located on the vertebrobasilar arterial system (RR: 4.4; CI 95%, 2.7-6.8), between 7 and 12 mm in size (RR: 3.3; CI 95%, 1.3-8.2) or larger (RR: 17; CI 95%, 8-36.1), multilobular and had a ratio of depth to width greater than 3.4 (risk x 20). A family history of UIA would constitute a major rupture risk (two to seven times that of spontaneous UIA). Other factors related to UIA rupture include arterial hypertension (RR: 1.46; CI 95%, 1.01-2.11) and smoking (RR: 3.04; CI 95%, 1.21-7.66). After microsurgical exclusion, the morbidity and mortality rates were calculated as 9 and 1.5%, respectively. Microsurgical risk factors were age (32% > 65 years), and factors related to the UIA itself and surgery, such as size (14% > 15 mm), location, presence of atherosclerosis and difficulty of surgical clip application. The incidence of rupture after microsurgical exclusion was estimated to be 0.2% per year, and complete microsurgical exclusion was achieved in 90% of patients. A randomised study of microsurgical exclusion of UIA would offer further proof of our therapeutic hypotheses. PMID- 18417220 TI - Application of in vitro neurotoxicity testing for regulatory purposes: Symposium III summary and research needs. AB - Prediction of neurotoxic effects is a key feature in the toxicological profile of many compounds and therefore is required by regulatory testing schemes. Nowadays neurotoxicity assessment required by the OECD and EC test guidelines is based solely on in vivo testing, evaluating mainly effects on neurobehavior and neuropathology, which is expensive, time consuming and unsuitable for screening large number of chemicals. Additionally, such in vivo tests are not always sensitive enough to predict human neurotoxicity and often do not provide information that facilitates regulatory decision-making processes. Incorporation of alternative tests (in vitro testing, computational modelling, QSARs, grouping, read-across, etc.) in screening strategies would speed up the rate at which compound knowledge and mechanistic data are available and the information obtained could be used in the refinement of future in vivo studies to facilitate predictions of neurotoxicity. On 1st June 2007, the European Commission legislation concerning registration, evaluation and authorisation of chemicals (REACH) has entered into force. REACH addresses one of the key issues for chemicals in Europe, the lack of publicly available safety data sheets. It outlines a plan to test approximately 30,000 existing substances. These chemicals are currently produced in volumes greater than 1ton/year and the essential data on the human health and ecotoxicological effects are lacking. It is estimated that approximately 3.9 million test animals (including 2.6 million vertebrates) (Hartung T, Bremer S, Casati S, Coecke S, Corvi R, Fortnaer S, et al. ECVAM's response to the changing political environment for alternatives: consequences of the European Union chemicals and cosmetics policies. ATLA 2003;31:473-81) would be necessary to fulfill the requirements of REACH if the development and establishment of alternative methods is not accepted by regulatory authorities. In an effort to reduce animal use and testing costs within this tonnage band, the European Commission has advocated the use of alternative approaches. Neurotoxicity testing is not directly addressed within REACH, however when alerts are observed based on organ specific toxicity studies then neurotoxicity assessment has to be performed. This session at the 11th International Neurotoxicology Association Meeting provided a forum to openly discuss and debate the potential of in vitro testing strategies that could be relevant for neurotoxicity evaluation in the context of regulatory requirements. The EU FP6 project A-Cute-Tox was presented as an example of a possible in vitro testing strategy for prediction of human acute systemic toxicity. Other presentations focused on the characterization of the available in vitro models (cell lines and primary culture) and neuronal specific endpoints, with a special emphasis on electrical activity, metabonomics and modulation of vesicular neurotransmitter release as possible neuronal endpoints relevant for in vitro neurotoxicity testing. Finally, it was underlined that in vitro systems (strategies) that have the potential to be applied for neurotoxicity assessment have to be formally validated under standardised conditions that have been recognised by national and international validation bodies. PMID- 18417222 TI - Adult bone marrow-derived mononuclear cells expressing chondroitinase AC transplanted into CNS injury sites promote local brain chondroitin sulphate degradation. AB - Injury to the CNS of vertebrates leads to the formation of a glial scar and production of inhibitory molecules, including chondroitin sulphate proteoglycans. Various studies suggest that the sugar component of the proteoglycan is responsible for the inhibitory role of these compounds in axonal regeneration. By degrading chondroitin sulphate chains with specific enzymes, denominated chondroitinases, the inhibitory capacity of these proteoglycans is decreased. Chondroitinase administration involves frequent injections of the enzyme at the lesion site which constitutes a rather invasive method. We have produced a vector containing the gene for Flavobacterium heparinum chondroitinase AC for expression in adult bone marrow-derived cells which were then transplanted into an injury site in the CNS. The expression and secretion of active chondroitinase AC was observed in vitro using transfected Chinese hamster ovarian and gliosarcoma cells and in vivo by immunohistochemistry analysis which showed degraded chondroitin sulphate coinciding with the location of transfected bone marrow-derived cells. Immunolabelling of the axonal growth-associated protein GAP-43 was observed in vivo and coincided with the location of degraded chondroitin sulphate. We propose that bone marrow-derived mononuclear cells, transfected with our construct and transplanted into CNS, could be a potential tool for studying an alternative chondroitinase AC delivery method. PMID- 18417221 TI - Higher blood harmane (1-methyl-9H-pyrido[3,4-b]indole) concentrations correlate with lower olfactory scores in essential tremor. AB - BACKGROUND: Harmane (1-methyl-9H-pyrido[3,4-b]indole), a neurotoxin, may be an environmental risk factor for essential tremor (ET). Harmane and related chemicals are toxic to the cerebellum. Whether it is through this mechanism (cerebellar toxicity) that harmane leads to ET is unknown. Impaired olfaction may be a feature of cerebellar disease. OBJECTIVE: To determine whether blood harmane concentrations correlate with olfactory test scores in patients with ET. METHODS: Blood harmane concentrations were quantified using high performance liquid chromatography. Odor identification testing was performed with the University of Pennsylvania Smell Identification Test (UPSIT). RESULTS: In 83 ET cases, higher log blood harmane concentration was correlated with lower UPSIT score (rho=-0.46, p<0.001). 25/40 (62.5%) cases with high log blood harmane concentration (based on a median split) had low UPSIT scores (based on a median split) vs. 12/43 (27.9%) ET cases with low log blood harmane concentration (adjusted odd ratios (OR) 4.04, 95% confidence intervals (CI) 1.42-11.50, p=0.009). When compared with the low log blood harmane tertile, the odds of olfactory dysfunction were 2.64 times higher in cases in the middle tertile and 10.95 times higher in cases in the high tertile. In 69 control subjects, higher log blood harmane concentration was not correlated with lower UPSIT score (rho=0.12, p=0.32). CONCLUSIONS: Blood harmane concentrations were correlated with UPSIT scores in ET cases but not controls. These analyses set the stage for postmortem studies to further explore the role of harmane as a cerebellar toxin in ET. PMID- 18417223 TI - Disturbed holistic processing in autism spectrum disorders verified by two cognitive tasks requiring perception of complex visual stimuli. AB - Central coherence is a key concept in research on autism spectrum disorders (ASD). It refers to the process in which diverse information is integrated and higher meaning is constructed in context. A malfunction in this process could result in abnormal attention to partial information in preference to the whole. To verify this hypothesis, we studied the performance of two visual tasks by 10 patients with autistic disorder or Asperger's disorder and by 26 (experiment 1) or 25 (experiment 2) normal subjects. In experiment 1, the subjects memorized pictures, some pictures with a change related to the main theme (D1) and others with a change not related to the main theme (D2); then the same pictures were randomly presented to the subjects who were asked to find the change. In experiment 2, the subjects were presented pictures of a normal (N) or a Thatcherized (T) face arranged side by side inversely (I) or uprightly (U) and to judge them as the same or different. In experiment 1, ASD subjects exhibited significantly lower rates of correct responses in D1 but not in D2. In experiment 2, ASD subjects exhibited significantly longer response times in NT-U but not in TN-I. These results showed a deficit in holistic processing, which is consistent with weak central coherence in ASD. PMID- 18417224 TI - The crucial role of IL-2/IL-2RA-mediated immune regulation in the pathogenesis of type 1 diabetes, an evidence coming from genetic and animal model studies. AB - Interleukin-2 (IL-2) plays an established role in T-cell regulation through binding to the high-affinity IL-2 receptor (IL-2R). The alpha-chain encoded by the IL2RA (CD25) gene is a substantial component of the high-affinity receptor molecule highly expressed by activated T lymphocytes. Recently, a strong evidence was obtained for the involvement of IL-2RA in conferring susceptibility to type 1 diabetes (T1D). Significant association with T1D was also found in the region on chromosome 4q27 containing the IL2 gene and homologous to the susceptibility locus idd3 in non-obese diabetic (NOD) mice, an animal model for human T1D. Here we focus on the discussion of these new findings suggesting for a crucial role of IL-2/IL-2RA-mediated regulatory mechanisms in preventing T1D. The non-redundant role of IL-2 and its receptor in etiology of T1D could be particularly attributable to the regulation of CD4+ CD25+ regulatory T cells, whose function is critical in maintaining immune homeostasis. PMID- 18417225 TI - Interleukin-17 in transverse myelitis and multiple sclerosis. AB - CSF IL-6 is elevated in transverse myelitis (TM) and predicts disability. Since IL-17 regulates cytokines (TNFalpha, IL-1beta and IL-6) known to stimulate IL-6 production by astrocytes, we sought to determine whether IL-17 was increased in TM and MS compared to healthy controls (HC) and other neurologic diseases (OND). IL-17 and IL-6 levels were measured in stimulated peripheral blood mononuclear cell (PBMC) supernatants from HC, MS, TM and OND. IL-17 was increased in TM compared to HC, MS, and OND (mean pg/ml+/-standard error; HC: 36.1+/-11.7, MS: 89.4+/-23.3, TM: 302.6+/-152.5, OND: 41.2+/-13.0, p=0.01). IL-6 was increased in TM relative to MS and HC (HC: 2624 pg/ml+/-641, MS: 6129+/-982, TM: 12,536+/ 2657, OND: 6920+/-1801, p<0.002). MS patients with early disease (<2 years) also had increased levels of IL-17 (p<0.04) and IL-6 (p<0.05). Cytokine neutralization experiments demonstrated that IL-6 was the main inducer of astrocyte IL-6 production. We conclude that IL-17 and IL-6 production from PBMC in TM and early MS are increased and induce astrocyte IL-6 production through IL-6. PMID- 18417226 TI - Hypoxia ischemia affects ultrasonic vocalization in the neonatal rat. AB - Perinatal hypoxic-ischemic encephalopathy results in a spectrum of pathologies related to the degree of initial infarct and environmental factors, including maternal interactions. Infants actively influence their environment by crying; rat pups produce ultrasonic vocalizations (USVs). Our study observed that ischemic pups engage in less time producing USVs and make fewer USVs overall, with male ischemic pups experiencing reductions in more categories than females. Future studies should consider whether alterations in mother-pup interactions result from these reductions. PMID- 18417227 TI - Long-term consequences of soman poisoning in mice Part 1. Neuropathology and neuronal regeneration in the amygdala. AB - To date, studies on soman-induced neuropathology mainly focused on the hippocampus, since this brain region is a well-delimited area with easily detectable pyramidal neurons. Moreover, the hippocampus is severely damaged after soman exposure leading to a substantial alteration of behavioral mnemonic processes. The neuropathology described in the hippocampus, however, and its behavioral consequences cannot be extrapolated to all other limbic damaged brain areas such as the amygdala. Accordingly, in this inaugural paper, using hemalun phloxin staining and NeuN immunohistochemistry, the number of damaged and residual healthy neurons was quantified in the amygdala in mice over a 90-day period after soman injection (1.2LD(50) of soman). On post-soman day 1, a moderate neuronal cell death (about 23% of the whole neurons) was evidenced. In parallel, a large quantity of degenerating neurons (about 36% of the whole neurons) occurred in this brain region and survived from post-soman day 1 to day 15. The death of these damaged neurons was initiated on post-soman day 30, and ended on post-soman day 90. Concomitantly, as quantified by NeuN immunohistochemistry, a clear neuronal regeneration was demonstrated in the amygdala of soman-poisoned mice between 60 and 90 days after neurotoxicant exposure. In the companion paper (see part 2), the possible effects of both long term neuropathology and delayed neuronal regeneration were evaluated on amygdala driven emotional processes. PMID- 18417228 TI - Disruption of the Plasmodium berghei 2-Cys peroxiredoxin TPx-1 gene hinders the sporozoite development in the vector mosquito. AB - To investigate the physiologic role of cytosolic 2-Cys peroxiredoxin of Plasmodium berghei (PbTPx-1), we infected the vector mosquito Anopheles stephensi with a parasite carrying a targeted knockout of pbtpx-1 (Prx-KO). The number of Prx-KO midgut oocysts at 14-15 days post-feeding (pf) was comparable to that of the parent strain (WT); however, the numbers of sporozoites that formed in midgut oocysts and accumulated in the salivary gland of Prx-KO-infected mosquitoes by 21 days pf were decreased to 10-20% and 3-10%, respectively, of those values in WT infected mosquitoes. A higher frequency of DNA strand breaks was detected in Prx KO oocysts than in WT oocysts. Sporozoites carrying the targeted disruption had reduced infectivity in mice; however, the knockout did not affect the ability of the sporozoite to reach the liver parenchyma and initiate exo-erythrocytic form (EEF) development. TPx-1 may be involved in development during exponentially multiplying stages, such as sporozoites and EEF. PMID- 18417229 TI - Veterinary public health: past success, new opportunities. AB - Animal diseases are known to be the origin of many human diseases, and there are many examples from ancient civilizations of plagues that arose from animals, domesticated and wild. Records of attempts to control zoonoses are almost as old. The early focus on food-borne illness evolved into veterinary medicine's support of public health efforts. Key historical events, disease outbreaks, and individuals responsible for their control are reviewed and serve as a foundation for understanding the current and future efforts in veterinary public health. Animal medicine and veterinary public health have been intertwined since humans first began ministrations to their families and animals. In the United States, the veterinary medical profession has effectively eliminated those major problems of animal health that had serious public health ramifications. These lessons and experiences can serve as a model for other countries. Our past must also be a reminder that the battle for human and animal health is ongoing. New agents emerge to threaten human and animal populations. With knowledge of the past, coupled with new technologies and techniques, we must be vigilant and carry on. PMID- 18417230 TI - Is managed care restraining the adoption of technology by hospitals? AB - As health care costs increase, cost-control mechanisms become more widespread and it is crucial to understand their implications for the health care market. This paper examines the effect that managed care activity (based on the aim to control health care expenditure) has on the adoption of technologies by hospitals. We use a hazard rate model to investigate whether higher levels of managed care market share are associated with a decrease on medical technology adoption during the period 1982-1995. We analyze annual data on 5390 US hospitals regarding the adoption of 13 different technologies. Our results are threefold: first, we find that managed care has a negative effect on hospitals' technology acquisition for each of the 13 medical technologies in our study, and its effect is stronger for those technologies diffusing in the 1990s, when the managed care sector is at its largest. If managed care enrollment had remained at its 1984 level, there would be 5.3%, 7.3% and 4.1% more hospitals with diagnostic radiology, radiation therapy and cardiac technologies, respectively. Second, we find that the rise in managed care leads to long-term reductions in medical cost growth. Finally, we take into account that profitability analysis is one of the main dimensions considered by hospitals when deciding about the adoption of new technologies. In order to determine whether managed care affects technologies differently if they have a different cost-reimbursement ratio (CRR), we have created a unique data set with information on the cost-reimbursement for each of the 13 technologies and we find that managed care enrollment has a considerably larger negative effect on the adoption of less profitable technologies. PMID- 18417231 TI - WST-1-based cell cytotoxicity assay as a substitute for MTT-based assay for rapid detection of toxigenic Bacillus species using CHO cell line. AB - Bacillus cereus continues to be one of the important foodborne pathogens due to its ability to produce various heat-labile and -stable toxins. Several methods have been developed to assess the pathogenicity of the B. cereus strains; however, most of these take more than 2-3 days to provide confirmatory results. In this study we standardized a one-step cytotoxicity assay using WST-1 (4-[3-(4 iodophenyl)-2-(4-nitrophenyl)-2H-5-tetrazolio]-1,3-benzene disulfonate) and compared with the traditional MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide)-based assay for rapid detection of cytotoxic Bacillus spp. using Chinese hamster ovary (CHO) cell line. Crude toxin preparations from 50 isolates of Bacillus spp. were exposed to CHO cell line for 1 h or 24 h and the cytotoxicity was determined by using WST-1 and MTT-based methods. Most B. cereus strains and some strains of other Bacillus species from our collection or from food sources showed comparably high cytotoxicity using either of the methods (P=0.81); however, WST-1 assay provided results in only 3 h while MTT assay in 44 52 h. A positive correlation (R2=0.93) between WST-1 and MTT assays strongly suggests that the WST-1-based cytotoxicity assay could be used as an alternative method to MTT assay for rapid (3 h) confirmation of toxigenic Bacillus species in foods prior to their retail distribution or consumption. PMID- 18417232 TI - An anthropomorphic phantom study of visualisation of surgical clips for partial breast irradiation (PBI) setup verification. AB - Surgical clips were investigated for partial breast image-guided radiotherapy (IGRT). Small titanium clips were insufficiently well visualised. Medium tantalum clips were best for megavoltage IGRT and small tantalum clips were best for floor mounted kilovoltage IGRT (ExacTrac). Both small tantalum and medium titanium clips were suitable for isocentric kilovoltage IGRT. PMID- 18417233 TI - Need for objective patient selection criteria for palliative radiotherapy in advanced head and neck cancer. PMID- 18417235 TI - Salivary alpha amylase and cortisol responses to different stress tasks: impact of sex. AB - Neuro-endocrine markers such as salivary alpha amylase (sAA) and cortisol (CORT) play an important role in establishing human responses to stressful events. Whereas sAA levels reflect sympathetic system activity, salivary cortisol appears to be a valid measure for HPA axis activity. Although many studies looked at either sAA or CORT responses in reaction to stress, work still has to be done to look at the way these systems interact, especially when both systems are activated. Additionally, sex effects in CORT responses have been investigated relatively often, but possible sex differences in sAA levels and responses, or the way both systems interact has not been the focus of sufficient studies to yield a univocal conclusion. In this study we presented a group of healthy participants (n=80) with two mildly stressful tasks, consisting of an aversive picture rating task and a cold pressor stress (CPS) task. The second task was compared with a control task. We expected a rise in sAA level in response to the first task and sAA as well as CORT responses on the second task and explored the interaction between the two responses. Results indicate that sAA is indeed a sensitive marker in both psychologically and physically induced arousal paradigms, whereas a cortisol response was only observed in the CPS task. Men had higher sAA levels than women during the complete course of the study, but men and women were comparable in their responsivity to the tasks. No strong correlations between sAA and CORT responses were found. PMID- 18417236 TI - Taxometric analysis of biceps femoris EMG following electrocutaneous stimulation over the sural nerve: determining the latent structure of the nociceptive flexion reflex (NFR). AB - The nociceptive flexion reflex (NFR) is a polysynaptic withdrawal reflex typically assessed from biceps femoris electromyogram (EMG) following noxious stimulation of the ipsilateral sural nerve. Electrophysiological evidence suggests the reflex is elicited following the activation of small diameter A delta afferents. As a result, the NFR is assumed to be a categorically distinct construct that emerges from EMG activity only following nociceptor activation. Despite the widespread use of the NFR in pain research, there has been little attempt to verify the latent structure of the NFR. The present study used "coherent cut kinetics" taxometric analyses to examine whether the latent structure of biceps femoris EMG reflects the taxonic structure that would be predicted from electrophysiological evidence. To achieve this end, preliminary analyses first compared different methods of scoring NFR magnitude. Results suggested the presence of a taxon in the covariance of biceps femoris EMG and stimulus intensity that is likely to be the NFR. Furthermore, preliminary analyses suggested the best method of scoring NFR magnitude was using Cohen's d. Implications of these results are discussed. PMID- 18417237 TI - Cardiovascular fitness and executive control during task-switching: an ERP study. AB - Cardiovascular fitness recently has been linked to executive control function in older adults. The present study examined the relationship between cardiovascular fitness and executive control in young adults using event-related potentials (ERPs). Participants completed a two-part experiment. In part one, a graded exercise test (GXT) was administered using a cycle ergometer to obtain VO(2)max, a measure of maximal oxygen uptake. High-fit participants had VO(2)max measures at or above the 70th percentile based on age and sex, and low-fit participants had VO(2)max measures at or below the 30th percentile. In part two, a task switching paradigm was used to investigate executive control. Task-switching trials produced slower response times and greater amplitude for both the P3a and P3b components of the ERP relative to a non-switch trial block. No ERP components varied as a function of fitness group. These findings, combined with results from previous research, suggest that the relationship between greater cardiovascular fitness and better cognitive function emerges after early adulthood. PMID- 18417238 TI - Carotenoid accumulation in bacteria with enhanced supply of isoprenoid precursors by upregulation of exogenous or endogenous pathways. AB - Carotenoids are isoprenoid pigments of industrial and nutritional interest. Although they are produced in non-carotenogenic Escherichia coli engineered with the appropriate biosynthetic genes, only a limited pool of their metabolic precursors is available in these bacteria. We have compared the production of carotenoids (lycopene) in strains in which the supply of precursors was enhanced either by upregulating the endogenous pathway via overexpression of deoxyxylulose 5-phosphate synthase (DXS) or by incorporating an exogenous MVA+ operon. In strains expressing DXS under the control of a leaky IPTG-inducible promoter, lycopene accumulation was increased up to 8-fold in the absence of inducer. Addition of IPTG, however, negatively affected lycopene production. Although induction of too high levels of the MVA+ operon enzymes also appeared to cause interference with cell metabolism, supplementation with mevalonate (to be metabolized into carotenoid precursors) resulted in a 10-fold increase in lycopene levels in cells with a near wild-type background. An additional 2-fold increase (up to 228mg/l) was obtained using an engineered BL21 strain. These results confirm that the MVA+ pathway is most convenient to upregulate the production of carotenoids (lycopene) production in E. coli but that factors other than precursor supply should be considered for high pigment accumulation levels. PMID- 18417239 TI - Characterisation of the first complete genome sequence of the roe deer (Capreolus capreolus) papillomavirus. AB - The complete genomic DNA of a novel roe deer (Capreolus capreolus) papillomavirus (CcPV1) was amplified and sequenced from fibropapillomatous skin lesions of a Hungarian roe deer. Viral DNA was detected by a pair of degenerate primers and the remaining genomic sequence was amplified by a long-template high-fidelity PCR and sequenced. The CcPV1 genome was 8032 bp long and contained open reading frames (ORFs) typical for Delta-papillomaviruses (E6, E7, E1, E2, E4, E5, E9, L2, and L1) and a 799 bp long untranslated regulatory region (URR). Phylogenetic analysis based on the 3861 bp long concatenated sequence of the E1-E2-L2-L1 ORFs and on separate alignments of all major ORFs using both neighbour-joining and maximum parsimony methods placed CcPV1 on a distinct branch between Ovine papillomavirus 1 and the other deer papillomaviruses within the Delta papillomavirus genus, although pairwise nucleotide alignments of L1 ORF sequences determined highest identities with European Elk Papillomavirus (71.2%) and Reindeer Papillomavirus (70.3%). PMID- 18417240 TI - Preparing polymer-based sustained-release systems without exposing proteins to water-oil or water-air interfaces and cross-linking reagents. AB - We report a method to load proteins into polymer-based sustained-release systems without exposing them to water-oil or water-air interfaces, factors known to denature proteins. By dispersing a dextan solution containing a protein into a PEG solution containing small amount of alginate, a stable aqueous-aqueous "emulsion" was formed. The poly-anionic alginate generated a diffuse double layer around each dextran droplet to prevent them from contacting with each other and fusing to a block phase. Proteins distributed in the stabilized dextran droplets by preferential partition favoring dextran. Freeze-drying this emulsion resulted in protein-loaded dextran particles, 1-2 microm in diameter and 1.6 g/cm(3) in density. The particles were harvested by washing the lyophilized powder using organic solvents to remove the PEG continuous phase. An activity assay of encapsulated beta-galactosidase indicated that protein activity was preserved during the particle-forming process including the step of sonicating the particles in organic solvents. The dextran particles also improved release profile and integrity of proteins when encapsulated in degradable polymer sustained-release systems. The aqueous-aqueous emulsion offers a convenient way to prepare solvent-resistant protein-polysaccharide particles that can easily be incorporated in a variety of polymer-based pharmaceutical dosage forms and medical devices such as microspheres, scaffolds and drug-eluting stents for sustained-release protein delivery. PMID- 18417241 TI - Analysis of muscle microcirculation in advanced diabetes mellitus by contrast enhanced ultrasound. AB - AIMS: Contrast enhanced ultrasound (CEUS) was recently established to quantify perfusion deficits in peripheral arterial disease (PAD). However, this approach was not suitable to assess microangiopathy of skeletal muscle, a major contributor to PAD in diabetic patients. We hypothesized that an optimized methodology would detect impaired microcirculation. METHODS: Ten patients with advanced diabetes mellitus (mean diabetes duration 21 years), 10 PAD patients, and 10 control subjects were enrolled consecutively. The arrival times of the contrast agent Sonovue after intravenous injection were assessed selectively in a small artery, muscle tissue and a muscle vein of the calf muscle. Contrast transit times (CTTs) were calculated as the differences between arrival times. RESULTS: The median CTT for artery-vein was significantly higher in the diabetes group (43 s) than in the PAD (22 s, p=0.007) and control groups (11 s, p<0.001, no value overlap). CTTs for artery-muscle and muscle-vein were shorter with highest median values in the diabetes group. CONCLUSIONS: We validated improved CEUS as consistent method to detect changes in the microvascular bed. This method may become a valuable tool to quantify impaired microcirculation in diabetes and help to improve patient care. PMID- 18417242 TI - Cardiovascular events and insulin therapy: a retrospective cohort analysis. AB - To investigate the impact of insulin plus other risk factors on cardiovascular disease (CVD) events in patients with type 2 diabetes (T2D), we conducted a retrospective study among patients with T2D from a U.S. managed care plan (n=342,692). Date of first CVD event was defined as index date. For patients without events, index date was date of last-observed claim. CVD event rates were calculated; odds of CVD event were compared for patients with/without insulin use. Events analyzed included stroke, MI, other cardiac/cerebrovascular events. Among insulin group (n=14,167), 22 patients/1000 patient-years experienced a CVD event, compared with 19/1000 patient-years in non-insulin group (n=328,077). Adjusting for risk factors and comorbidities, odds of event for insulin group compared to non-insulin group was 0.66, ranging from 25% lower for patients aged 65+ to 42% lower for patients aged 31-45. Insulin treatment of T2D was associated with reduced risk of CVD events compared to other or no therapy. PMID- 18417243 TI - The effect of continuous subcutaneous glucose monitoring (CGMS) versus intermittent whole blood finger-stick glucose monitoring (SBGM) on hemoglobin A1c (HBA1c) levels in Type I diabetic patients: a systematic review. AB - BACKGROUND AND PURPOSE: Continuous glucose monitoring (CGMS) is a relatively new technology that measures interstitial glucose every 5 min for 72 h. The resulting profile provides a more comprehensive measure of glycemic excursions than intermittent self-blood finger-stick glucose monitoring (SBGM) and thus could potentially improve diabetes control. We performed a meta-analysis of randomized controlled trials comparing CGMS and SBGM in Type 1 diabetic patients. Our aim was to determine whether CGMS leads to better hemoglobin A1c (HBA1c) levels, a marker of long-term vascular risk. METHODS: Randomized controlled trials comparing CGMS and SBGM in Type 1 diabetic patients were identified using both manual and electronic searches of the literature in MEDLINE, EMBASE, PUBMED and Cochrane Central Registry of Controlled Trials from 1996 to March 2007. Relevant studies were independently selected by two reviewers, who also extracted data on study design, quality and effect on HBA1c levels. Data from all trials were pooled using a random effects model. RESULTS: Seven studies with a total of 335 patients fulfilled the inclusion criteria. Five studies were confined to the pediatric population (age<18 years). Study duration varied from 12 to 24 weeks. Compared with SBGM, CGMS was associated with a non-significant reduction in HBA1c (0.22%; 95% CI: -0.439% to 0.004%, p=0.055). CONCLUSIONS: There is insufficient evidence to support the notion that CGMS provides a superior benefit over SBGM in terms of HBA1c reduction. There was some indication of improved detection of asymptomatic nocturnal hypoglycemia in the CGMS group. PMID- 18417244 TI - Give me a break! Informal caregiver attitudes towards respite care. AB - BACKGROUND/OBJECTIVE: Because informal health care is now recognized to be indispensable to health care systems, different forms of respite care have been developed and publicly funded that supposedly alleviate caregivers' perceived burdens and help prolong the care giving task. Nonetheless, the use of respite care services is low even among substantially strained caregivers. To throw light on this low usage, this paper explores the associations between attitudes towards respite care, characteristics of the care giving situation, and the need and use of respite care. METHOD: The survey, administered to a sample of 273 informal caregivers, addressed caregiver, care recipient, and care giving situation characteristics, as well as the familiarity and use of respite care services. It also included a sub-set of 12 statements eliciting attitudes towards respite care from an earlier study [Van Exel NJA, De Graaf G, Brouwer WBF. Care for a break? An investigation of informal caregivers' attitudes toward respite care using Q methodology. Health Policy 2007;83(2/3):332-42]. Associations between variables were measured using univariate statistics and multinomial logistic regression. RESULTS: We found three caregiver attitudes, distributed fairly equally in the sample, that are apparently associated with caregiver educational level, employment status, health and happiness, as well as care recipient gender, duration and intensity of care giving, relationship, co-residence, need for surveillance, and subjective burden and process utility of care giving. However, the relation between attitude and familiarity with and use of respite care services is ambiguous. CONCLUSIONS: Although further exploration is needed of the mix of Q-methodology and survey analysis, the overall results indicate that a considerable portion of the caregiver population needs but does not readily ask for support or respite care. This finding has important policy implications in the context of an ageing population. PMID- 18417245 TI - Towards an integrated system for planning and assisting maxillofacial orthognathic surgery. AB - Computer-assisted maxillofacial orthognathic surgery is an emerging and interdisciplinary field linking orthognathic surgery, remote signal engineering and three-dimensional (3D) medical imaging. Most of the computational solutions already developed make use of different specialized systems which introduce difficulties both in the information transfer from one stage to the others and in the use of such systems by surgeons. Trying to address such issue, in this work we present a common computer-based system that integrates proposed modules for planning and assisting the maxillofacial surgery. With that we propose to replace the current standard orthognathic preoperative planning, and to bring information from a virtual planning to the real operative field. The system prototype, including three-dimensional cephalometric analysis, static and dynamic virtual orthognathic planning, and mixed reality transfer of information to the operation room, is described and the first results obtained are presented. PMID- 18417246 TI - Treatment outcome in performance status 2 advanced NSCLC patients administered platinum-based combination chemotherapy. AB - BACKGROUND: There is no consensus regarding chemotherapy to patients with advanced NSCLC (ANSCLC) and performance status (PS) 2. Using data from a national multicenter study comparing two third-generation carboplatin-based regimens in ANSCLC patients, we evaluated the outcome of PS 2 patients. PATIENTS AND METHODS: The 123 PS 2 patients were compared to 309 PS 0/1 patients regarding survival, quality of life (QOL) and treatment toxicity. RESULTS: PS 2 patients had lower haemoglobin, lower global QOL and more pain, nausea/vomiting and dyspnea at inclusion. 68% of PS 2 patients received three chemotherapy courses vs. 85% in the PS 0/1 group (P<0.01). Median and 1-year survival were lower in the PS 2 group, 4.5 vs. 8.9 months and 10% vs. 37% (P<.01). More PS 2 patients needed blood transfusions (P=0.03) and hospitalization (P<0.01). In contrast, PS 2 patients had better relief of pain and dyspnea, and tended to a better global QOL and did not experience more leucopoenia, infections or bleeding. CONCLUSIONS: Despite shorter survival, treatment toxicity was acceptable and PS 2 patients achieved better improvement of pain and dyspnea and tended to better global QOL when compared to PS 0/1 patients. PMID- 18417247 TI - Fates and osteogenic differentiation potential of human mesenchymal stem cells in immunocompromised mice. AB - Human mesenchymal stem cells (hMSCs) from bone marrow were genetically marked by using a murine leukaemia virus construct encoding enhanced green fluorescent protein (eGFP). The marked cells were either directly implanted into the tibialis anterior muscle or introduced into a variety of other tissue sites in immunocompromised mice (NOD/SCID and C.B-17 SCID/beige) to investigate their fates and differentiation potentials. It was observed that the hMSCs survived for up to 12 weeks and showed site-specific morphological phenotypes. hMSCs delivered by intravenous injection were found mainly in the lungs and were detected rarely in other organs. Histomorphometry showed that, after implantation of hMSCs into the tibialis anterior muscle juxtaskeletally, the areas of reactive host callus formation at 1 and 2 weeks and of ectopic human bone formation at 1 week were significantly increased compared with the control group. Expression of eGFP and human RUNX2, alkaline phosphatase, osteocalcin, osteopontin, and collagen type I mRNAs were detected in mice implanted with the labelled hMSCs but not in sham treated samples. Active clearance of the reactive callus and ectopic calcified tissue by osteoclast-like tartrate-resistant acid phosphatase-positive cells was observed. We conclude that the eGFP-labelled hMSCs can survive and retain the potential to differentiate morphologically into a variety of apparent mesenchymal phenotypes in vivo. Absolute confirmation of differentiation capacity requires further study and is complicated by known possibilities of fusion of donor and host cells or limited transfer of genetic material. Nevertheless, the genetically marked hMSCs are shown to participate extensively in bone formation and turnover. Control of the host osteoclast/macrophage responses resulting in clearance of formed osteogenic tissue warrants further investigation to promote prolonged human osteogenesis in immunocompromised mice. Furthermore, any proposed general cytotherapeutic strategy for enhanced osteogenesis is likely to require supplementation of local bone-forming biological signals. PMID- 18417248 TI - Viva adhesion! A Mediterranean meeting on cell-matrix contacts and tissue invasion. PMID- 18417249 TI - A role for the podosome/invadopodia scaffold protein Tks5 in tumor growth in vivo. AB - Podosomes and invadopodia are electron-dense, actin-rich protrusions located on the ventral side of the cellular membrane. They are detected in various types of normal cells, but also in human cancer cells and in Src-transformed fibroblasts. Previously we have shown that the scaffold protein Tks5 (tyrosine kinase substrate 5) co-localizes to podosomes/invadopodia in different human cancer cells and in Src-transformed NIH-3T3 cells. Upon reduced expression of Tks5 podosome formation is decreased, which leads to diminished gelatin degradation in vitro in various human cancer cell lines. It is unclear, however, whether cancer cells need podosomes for tumor growth and metastasis in vivo. To test this idea, we evaluated the ability of Src-transformed NIH-3T3 cells, showing stable reduced expression of Tks5 and podosome formation (Tks5 KD), to form subcutaneous tumors in mice. We demonstrate that decreased expression of Tks5 correlated with reduced tumor growth at this site. In addition, we generated lung metastases from these cells following tail vein injection. The lungs of mice injected i.v. with the Tks5 KD showed smaller-sized metastases, but there was no difference in the number of lesions compared to the controls, indicating that podosomes may not be required for extravasation from the blood stream into the lung parenchyma. Independent of the microenvironment however, the reduced tumor growth correlated with decreased tumor vascularization. Our data potentially implicate a novel role of podosomes as mediators of tumor angiogenesis and support further exploration of how podosome formation and Tks5 expression contribute to tumor progression. PMID- 18417250 TI - Podosome-type adhesions and focal adhesions, so alike yet so different. AB - Cell-matrix adhesions are essential for cell migration, tissue organization and differentiation, therefore playing central roles in embryonic development, remodeling and homeostasis of tissues and organs. Matrix adhesion-dependent signals cooperate with other pathways to regulate biological functions such as cell survival, cell proliferation, wound healing, and tumorigenesis. Cell migration and invasion are integrated processes requiring the continuous, coordinated assembly and disassembly of integrin-mediated adhesions. An understanding of how integrins regulate cell migration and invasiveness through the dynamic regulation of adhesions is fundamental to both physiological and pathological situations. A variety of cell-matrix adhesions has been identified, namely, focal complexes, focal adhesions, fibrillar adhesions, podosomes, and invadopodia (podosome-type adhesions). These adhesion sites contain integrin clusters able to develop specialized structures, which are different in their architecture and dynamics although they share almost the same proteins. Here we compare recent advances and developments in the elucidation of the organization and dynamics of focal adhesions and podosome-type adhesions, in order to understand how such subcellular sites - though closely related in their composition - can be structurally and functionally different. The underlying question is how their respective physiological or pathological roles are related to their distinct organization. PMID- 18417251 TI - Identification of the insulin-responsive tyrosine phosphorylation sites on IRSp53. AB - The 53-kDa insulin receptor substrate protein (IRSp53) is part of a regulatory network that organises the actin cytoskeleton in response to stimulation by small GTPases, promoting formation of actin-rich cell protrusions such as filopodia and lamellipodia. It had been established earlier that IRSp53 is tyrosine phosphorylated in response to stimulation of the insulin and insulin-related growth factor receptors, but the consequences of tyrosine phosphorylation for IRSp53 function are unknown. Here, we have used a variety of IRSp53 truncation and point mutants to identify insulin-responsive tyrosine phosphorylation sites on IRSp53. We have found that the C-terminal half of IRSp53 (residues 251-521) undergoes tyrosine phosphorylation in response to insulin stimulation of the insulin beta receptor or epidermal growth factor stimulation via the epidermal growth factor receptor, and that the key residue for insulin receptor-mediated phosphorylation is tyrosine 310, located in a region between the N-terminal IRSp53/MIM homology domain (IMD, residue 1-250) and the central SH3 domain (residues 374-438) that is predicted to be natively unstructured. Mutation of tyrosine 310 to phenylalanine or glutamic acid abrogates the phosphorylation in response to insulin stimulation, but not in response to stimulation of the epidermal growth factor receptor. The N-terminal IMD, which mediates dimerisation of IRSp53, is required for efficient tyrosine phosphorylation downstream of either the insulin or epidermal growth factor receptor stimulation, yet does not appear to include a tyrosine-phosphorylated site itself. Thus, we have identified tyrosine 310 as a primary site of tyrosine phosphorylation in response to insulin signalling and we have shown that although IRSp53 is tyrosine phosphorylated in response to epidermal growth factor receptor signalling, tyrosine 310 is not crucial. Furthermore, the tyrosine phosphorylation status does not appear to affect the cell morphology and production of filopod-like structures upon expression of IRSp53. PMID- 18417252 TI - Selective regulation of the perinuclear distribution of glucose transporter 4 (GLUT4) by insulin signals in muscle cells. AB - Insulin regulates glucose transporter 4 (GLUT4) availability at the surface of muscle and adipose cells. In L6 myoblasts, stably expressed GLUT4myc is detected mostly in a perinuclear region. In unstimulated cells, about half of perinuclear GLUT4myc colocalizes with the transferrin receptor (TfR). Insulin stimulation selectively decreased the perinuclear colocalization of GLUT4myc with TfR determined by 3D-reconstruction of fluorescence images. Perinuclear GLUT4myc adopted two main distributions defined morphometrically as 'conical' and 'concentric'. Insulin rapidly reduced the proportion of cells with conical in favor of concentric perinuclear GLUT4myc distributions in association with the gain in surface GLUT4myc. Upon removal of insulin, the GLUT4myc perinuclear distribution and surface levels reversed in parallel. In contrast, hypertonicity (which like insulin elevates surface GLUT4myc) did not elicit perinuclear GLUT4myc redistribution. Insulin also caused redistribution of perinuclear vesicle-associated membrane protein-2 (VAMP2), without alteration of perinuclear TfR and VAMP3. Inhibitory mutants of phosphatidylinositol-3 kinase (Deltap85) or Akt substrate AS160 (AS160-4P) prevented insulin-mediated perinuclear GLUT4myc redistribution. Tetanus toxin expression did not prevent the perinuclear GLUT4myc redistribution, suggesting that redistribution is independent of GLUT4myc fusion with the plasma membrane. We propose that insulin causes selective, dynamic relocalization of perinuclear GLUT4myc and VAMP2 and perinuclear GLUT4myc redistribution is a direct target of insulin-derived signals. PMID- 18417253 TI - The association between food cravings and consumption of specific foods in a laboratory taste test. AB - This pilot study tested the relation between food cravings and food intake in the laboratory. Participants (n=91; mean BMI=35.1 kg/m2) completed the Food Craving Inventory to measure cravings for sweets, fats, carbohydrates, and fast food fats, and a taste test consisting of four foods (jelly beans, M&M's, regular potato chips, and baked low-fat potato chips). Thereafter, participants could eat the items ad libitum. Specific food cravings were significantly correlated with consumption of corresponding types of foods. The sweets scale correlated with M&M and jelly bean intake, but not chip intake. The fats scale correlated only with intake of regular potato chips. PMID- 18417254 TI - Who consumes functional foods and nutraceuticals in Canada? Results of cluster analysis of the 2006 survey of Canadians' demand for food products supporting health and wellness. AB - This research seeks to identify consumer segments related to consumption of functional food and nutraceutical products in Canada. The segments are differentiated by consumer receptivity to functional foods and nutraceuticals. In turn, receptivity is tied to attitudes, motivations and knowledge related to food/diet and connections with health. At one extreme, a segment emerges that is highly receptive to functional foods and nutraceuticals, and shows a keen interest in learning about foods that have health benefits. A second segment, conversely, has low receptivity, but demonstrates a higher degree of knowledge related to the relationship between food/diet and health. A key driver of receptivity towards functional foods and nutraceuticals appears to be perceptions of disease threat, which tend to increase with age. Pre-emptive use of messages relating to the preventative properties of these products does not seem to have a major impact on the receptivity of consumers. PMID- 18417255 TI - Effects of homocysteine on metabolic pathways in cultured astrocytes. AB - Homocysteine is an amino acid that is an important risk factor for several neurodegenerative diseases such as Alzheimer's and Parkinson's disease. Increased homocysteine levels induce neuronal cell death in a variety of neuronal types. However, very few studies have probed the effects of homocysteine in astrocytes. The present study investigated the effects of homocysteine on primary cultures of astrocytes by exposing astrocytes to 400 microM homocysteine for 20 h. Metabolic extracts of cells were prepared following a 4-h incubation in minimum medium with 5.5 mM [U-(13)C]glucose in the presence or absence of homocysteine and analysed using (13)C NMR. The expression level of pyruvate dehydrogenase kinase isoform 2 (PDK-2), NAD(P)H levels and mitochondrial membrane potential responses were investigated following culture with homocysteine. Metabolomic analysis was performed using (1)H NMR spectroscopy and pattern recognition analysis. Following incubation with homocysteine there was a significant decrease (48%) in the ratio of flux through pyruvate carboxylase (PC) and pyruvate dehydrogenase (PDH) which was due to an increased flux through PDH. In addition, homocysteine culture resulted in a significant reduction in PDK-2 protein expression. Following stimulation with glucose there was a significant increase in NAD(P)H levels and an impaired hyperpolarisation of the mitochondrial membrane in homocysteine treated cells. Metabolomic analysis showed that the most discriminating metabolites following homocysteine treatment were choline and hypotaurine. In summary, the results demonstrated that sub-lethal concentrations of homocysteine caused significant metabolic changes and altered mitochondrial function in primary cultures of astrocytes. PMID- 18417256 TI - Synthesis, Raman, FT-IR, NMR spectroscopic data and antimicrobial activity of mixed aza-oxo-thia macrocyclic compounds. AB - Mixed aza-oxo-thia macrocyclic ligands 1,3,5,11,13,15-hexaaza-6,10,16,20-tetraoxo 8,18-dithia-2,3,4:12,13,14-dipyridine cyclocosane (L(1)); 1,3,5,12,14,16-hexeaza 6,11,17,22-tetraoxo-8,9,19,20-tetrathia-2,3,4:13,14,15-dipyridine cyclodocosane (L(2)); 1,3,5,13,15,17-hexaaza-6,12,18,24-tetraoxo-9,21-dithia-2,3,4:14,15,16 dipyridine cyclotetracosane (L(3)) and 1,3,5,14,16,18-hexaaza-6,13,19,26-tetraoxo 9,10,22,23-tetrathia-2,3,4:15,16,17-dipyridine cyclohexacosane (L(4)) were synthesised. The structural features of the ligands have been studied by elemental analyses, Raman, IR, (1)H and (13)C NMR spectroscopy. The antimicrobial activities of the ligands were evaluated using disk diffusion method in dimethyl sulfoxide (DMSO) as well as the minimal inhibitory concentration (MIC) dilution method, against nine bacteria. The obtained results from disk diffusion method were assessed in side-by-side comparison with those of penicillin G, ampicillin, cefotaxime, vancomycin, ofloxacin, and tetracycline well known antibacterial agents. The results from dilution procedure were compared with gentamycin as antibacterial and nystatin as antifungal. The antifungal activities are reported on five yeast cultures namely Candida albicans, Kluyveromyces fragilis, Rhodotorula rubra, Debaryomyces hansenii, and Hanseniaspora guilliermondii, and the results are referenced with nystatin, Ketoconazole, and clotrimazole, commercial antifungal agents. In most cases, the compounds show broad-spectrum (Gram(+) and Gram(-) bacteria) activities that were more active or equipotent to the antibiotic and antifungal agents in the comparison tests. PMID- 18417257 TI - A cost-utility analysis in patients receiving enteral tube feeding at home and in nursing homes. AB - A cost-utility analysis was undertaken of enteral tube feeding (ETF) in patients with cerebrovascular accident (CVA). Mortality during ETF was established in nursing homes (n7007) or at home (n2888). Quality of life was measured (EuroQol) on a sub-sample (n25). Quality adjusted life years (QALYs) over 3 years and the cost/QALY were calculated. The cost/QALY (1 pound=1.482 euros; 30/06/05) for patients receiving ETF at home was 12,817 pounds (10,351 pounds-16,826 pounds using 95% CI for quality of life) and insensitive to the frequency of home visits (50-150% of the average frequency; 11,851 pounds-13,782 pounds), outcome of patients reverting to full oral feeding (0-100% survival; 11,023 pounds-14,440 pounds), and computed outcome of a 'control' group not given ETF (0.125-0.25 year survival; 12,991 pound-14,006 pounds). The cost/QALY in nursing homes (10,304 pounds-68,064 pounds) varied depending on the state contribution to non-medical costs and is above and below the threshold (30,000 pounds/QALY). The data suggest the cost-effectiveness of ETF in patients with CVA receiving ETF at home or in nursing homes, where the non-medical costs are paid privately, compares favourably with other interventions. The cost-effectiveness of ETF in nursing homes when the state pays all non-medical costs compares unfavourably with other treatments, raising economic, clinical, and ethical concerns. PMID- 18417258 TI - Delivery of vaccine peptides by rapid conjugation to baculovirus particles. AB - Baculoviruses deliver strong activation signals to dendritic cells and can promote potent immune responses. These properties can be harnessed to use baculovirus as an adjuvant and carrier particle for immunogenic peptides. In this study we use a chemical linker to couple peptides to the baculovirus Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV). Intranasal delivery of baculovirus coupled with immunogenic peptides to mice elicited antigen-specific IgG1 and IgG2a antibody. Furthermore, antigen-specific IgA was detected in the lung, and an IFN-gamma response was observed upon re-stimulation with antigen. We show that chemical coupling enables the rapid modification of AcMNPV, allowing multiple epitopes to be delivered simultaneously on a self-adjuvanting carrier particle. PMID- 18417259 TI - Development of a travelers' diarrhea vaccine for the military: how much is an ounce of prevention really worth? AB - Infectious diarrhea is one of the many threats to the deployed military, and given limited resources, a decision to pursue a vaccine acquisition strategy should be based on best evidence that weighs costs and benefits compared to alternatives. An economic model was developed to estimate the marginal cost to avert a duty day lost due to diarrhea for a vaccine acquisition strategy compared to current clinical management, for both multiplex and pathogen-specific vaccines. Vaccines against Campylobacter and enterotoxigenic Escherichia coli appeared to be more favorable than a Shigella vaccine. This model provides an evidence-based decision tool to support prioritization in vaccine development. PMID- 18417260 TI - Kikuchi disease: an unusual cause of cervical lymphadenopathy in an Afro Caribbean woman. AB - Cervical lymphadenopathy is often seen in maxillofacial surgery departments and many common causes have been identified. We discuss an unusual case of cervical lymphadenopathy and weight loss in an Afro-Caribbean patient. PMID- 18417261 TI - The hematologic manifestations of the antiphospholipid syndrome. AB - Various hematological pathologies have been described in association with antiphospholipid syndrome (APS). Thrombocytopenia is frequently found in APS patients, its incidence has ranged from 22-42% in different series, it is usually moderate (>50x10(9)/L) without clinical manifestation and requires no intervention. A high percentage of patients with isolated idiopathic thrombocytopenic purpura have antiphospholipid antibodies, however the pathogenetic role and the clinical importance of these antibodies in this condition is still not clear. Other hematological manifestations reported in association with APS include: bone marrow necrosis, and various thrombotic microangiopathic syndromes such as: thrombotic thrombocytopenic purpura, hemolytic-uremic syndrome, HELLP (hemolysis, elevated liver enzymes, low platelet) syndrome, and catastrophic APS. A high index of suspicion is needed for the early recognition and treatment of these conditions. PMID- 18417262 TI - Multidisciplinary approach of organic catatonia in children and adolescents may improve treatment decision making. AB - Catatonia is an infrequent but severe condition in young people. Organic diseases may be associated and need to be investigated though no specific recommendations and guidelines are available. We extensively reviewed the literature of all the cases of organic catatonia in children and adolescents from January 1969 to June 2007. We screened socio-demographic characteristics, organic diagnosis, clinical characteristics and treatment. We found 38 cases of children and adolescents with catatonia due to an organic condition. The catatonic syndrome occurred in 21 (57%) females and 16 (43%) males. The mean age of patients was 14.5 years (+/ 3.39) [range=7-18 years], and three died from their condition. The organic conditions included infectious diseases (N=10), neurological conditions (N=10), toxic induced states (N=12) and genetic conditions including inborn errors of metabolism (N=6). The onset was dominantly acute, and the clinical presentation most frequently stuporous. Although benzodiazepines were recommended as primary symptomatic treatment, they were rarely prescribed. In several cases, therapeutic approach was related to organic cause (e.g., plasma exchange in lupus erythematosus; copper chelators in Wilson's disease). Based on this review and on our own experience of catatonia in youth, we proposed a consensual and multidisciplinary diagnostic strategy to help practitioners to identify underlying organic diseases. PMID- 18417263 TI - Human percutaneous absorption of a direct hair dye comparing in vitro and in vivo results: implications for safety assessment and animal testing. AB - Although in vitro skin absorption studies often detect small residues of applied test material in the epidermis/dermis, it is uncertain whether the residue is within the living skin. We studied the dermal absorption of a hair dye hydroxyanthraquinone-aminopropyl methyl morpholinium methosulphate (HAM) in human skin in vivo and in vitro. In vivo, skin (back and scalp) received 0.5% HAM in a commercial formulation at 20microg/cm2 After 0.5 or 48h, skin was tape stripped, followed by cyanoacrylate biopsies (CAB). Sebum from scalp sites was collected for 48h. In vitro, skin was treated with 20mg/cm2 dye for 0.5h, penetration determined after 24h. In vivo, at 0.5h, total recovery (back) was 0.67microg/cm2 (tape strips+CAB). Fluorescence microscopy showed HAM in the hair follicle openings (HFO). At 0.5h, scalp tape strips contained 1.80microg/cm2, HFO 0.82microg/cm2. At 48h, HFO contained 0.21microg/cm2, sebum 0.80microg/cm2. In vivo, skin residues were in the non-living skin and eliminated via desquamation and sebum secretion. In vitro, the SC contained 1.50microg/cm2, epidermis/dermis 0.86microg/cm2, receptor fluid<0.04microg/cm2, a total of 0.90microg/cm2 was considered to be bioavailable. In vitro epidermis/dermis residues were nearly identical to those located in non-living skin in vivo. In conclusion, in vitro percutaneous penetration studies may produce seemingly bioavailable material , which raises the need for a Threshold of Skin Absorption (TSA) addressing a negligible dermal absorption in order to avoid unnecessary in vivo toxicity studies on substances that produce no significant human systemic exposure. PMID- 18417264 TI - Toxic effects of perinatal lead exposure on the brain of rats: involvement of oxidative stress and the beneficial role of antioxidants. AB - The aim of this study was to determine whether changes in the activities of antioxidant enzymes occur in the brain of lead-exposed rats (300mgPb/L in drinking water) and to investigate the potential benefit of the administration of some natural antioxidants (Zn 20mg/L+vitamins A 50.000U/L, C 2g/L, E 500mg/L and B6 500mg/L) during pregnancy and lactation. Lead exposure caused a significant increase in brain TBARS (23%) vs. control, whereas co-administration of antioxidants+lead was effective in reducing TBARS levels. The catalase activity in brain samples of the lead group was enhanced 99% vs. control, but no changes were found in the remainder of the groups. No statistically significant effect of lead and/or antioxidants in brain SOD activity was noted. Acid phosphatase activity was enhanced in both lead groups but no changes were found in alkaline phosphatase activity. Finally, a statistically significant decrease (-35%) of acetylcholinesterase activity was noted in the lead+antioxidants group. This study provides evidence of the beneficial role of antioxidants in early status of brain development in rats against lead exposure. PMID- 18417265 TI - Comment to "Di-(2-ethylhexyl) phthalate is without adjuvant effect in mice on ovalbumin" [Toxicology 244 (2008) 231-241]. PMID- 18417266 TI - Hepatic injuries of hexachloroethane smoke inhalation: the first analytical epidemiological study. AB - There has been no human epidemiological data regarding the hepatic injuries of hexachloroethane-zinc oxide (HC/ZnO) inhalation. This is the first epidemiological study to investigate whether HC/ZnO inhalation exposure can induce hepatic dysfunction in exposed soldiers. Twenty soldiers, exposed to high concentration of HC/ZnO smoke for 3-10 min in a narrow tunnel (0.6 m in width) during military training, were recruited as exposed group and they were divided into high-exposed group (n=10) and low-exposed group (n=10) by the distance from the explosion locale as a surrogate of exposure condition. Another 64 soldiers, not visiting the explosion areas, were recruited as referents. Venous blood was collected for liver function analyses. After log transformation of alanine aminotransferase (ALT) and adjustment for potential confounders, serum ALT in high-exposed soldiers was statistically significantly higher than those of referents for the 3 weeks following exposure. The serum ALT in low exposed soldiers was statistically significantly higher than those of referents at the 3rd week following exposure. The mean ALT levels also showed decreasing gradients by the distance from exposure locale. In addition, the proportions of abnormality on ALT (>40U/L) were also significantly different among three exposure conditions. Follow-up study showed that the hepatic dysfunction started from 1 to 2 weeks and peaked from 3rd to 5th week after exposure. ALT level was then returned to normal within 6-8 weeks after removing from HC/ZnO smoke exposure. No sequelas in hepatic dysfunction were found until 72 weeks follow-up. CONCLUSION: We concluded that inhalation of HC/Zn smoke can induce acute, dose-dependent and definite temporal relationship hepatic dysfunction. PMID- 18417267 TI - An investigation of factors influencing patients' satisfaction with new complete dentures using structural equation modelling. AB - OBJECTIVE: To investigate possible relationships between patient and clinical factors and patient satisfaction with new complete dentures. METHODS: Patients attending King's College London Dental Institute, Guy's Hospital, London, for new complete dentures were recruited. Relevant history was recorded and the edentulous mouth and existing dentures, where present, were examined at the beginning of treatment. The edentulous mouth, the previous and new dentures were re-examined at the first post-insertion visit. Three months after this visit, patients were sent a postal questionnaire to assess patient satisfaction. Data were analysed using structural equation modelling techniques. RESULTS: 723 patients were recruited. Significant relationships were found between quality of complete dentures, quality of residual alveolar, adaptability factors and patients' satisfaction with complete dentures. CONCLUSIONS: Quality of the mandibular residual alveolar ridges, retention and stability of the mandibular dentures, accuracy of reproduction of retruded jaw relationship and patient adaptability were powerful determinants of patients' satisfaction with new complete dentures. PMID- 18417268 TI - Associations among central nervous system serotonergic function and neuroticism are moderated by gender. AB - Serotonergic dysregulation is associated with negative affect. Plasma prolactin responses to a tryptophan enhancement challenge are used as a measure of central nervous system serotonergic activity. We examined prolactin responses to a tryptophan challenge as they relate to the personality domains of neuroticism, extraversion, openness, agreeableness, and conscientiousness. Participants were 67 volunteers. Regression models assessed peak prolactin response to intravenous tryptophan infusion as a predictor of neuroticism, extraversion, openness, agreeableness, and conscientiousness. Prolactinxgender product terms were included to examine moderation by gender. Models were adjusted for baseline levels of prolactin, age, and race. Gender moderated the association between N and prolactin level (p<.03). Higher levels of N were associated with decreased levels of prolactin responses in females, whereas the opposite was true for males. Remaining personality domains were not related to prolactin levels. Findings add to literature suggesting the serotonin system functions differently, in important ways, in males and females. PMID- 18417269 TI - Temperament, health-related behaviors, and autonomic cardiac regulation: the cardiovascular risk in young Finns study. AB - Temperament, as indicated by Cloninger's psychobiological model predicts coronary heart disease risk, but its association with autonomic cardiac regulation, a potential mediating mechanism, is unclear. We examined the associations between temperament traits and autonomic cardiac regulation in a resting situation in 798 women and 580 men derived from a population-based sample. After adjustment for age and sex, harm avoidance was associated with lower level of high-frequency (HF) variation, root mean square successive differences (RMSSDs), the percentage of successive R-R intervals>50 ms (pNN50) and higher heart rate (HR) (all p or =13 and a peak urinary flow rate (PFR) 5-13ml/s were randomly allocated to 4 treatment groups; patients with residual urine volume of >350ml were excluded. INTERVENTION: Patients received either CET at dosages of 5mg/wkx4, 10mg/2 wkx2 or 10mg/wkx4 or placebo. MEASUREMENTS: IPSS, PFR and mean uroflow, residual urinary volume, prostate volume, plasma testosterone, quality of life, and sexual function were evaluated over a total of 20 wk after randomization. RESULTS AND LIMITATIONS: Of 140 randomized patients, one patient did not complete treatment, 5 others dropped out off-treatment, before week 12 evaluation of the primary end point. In all CET groups a rapid improvement in mean IPSS was obtained, with a peak effect of -5.4 to -5.9 (placebo: -2.8). After all dosages of CET given, changes from baseline and differences to placebo were statistically significant up to week 20. Similarly, secondary parameters showed a significant, rapid, and persistent improvement for all CET dosages. All dosage regimens were well tolerated. The study evaluated a single treatment course only; further studies with repeated treatment courses will be required to establish a dose regimen for long-term disease management. CONCLUSIONS: At all dosage regimens tested, CET was safe and effective in patients with symptomatic BPH, with a trend towards a more rapid onset of effect for the CET 10mg/wkx4 regimen. Response persisted up to the end of follow-up, 16 wk after the last dose. PMID- 18417276 TI - B7-H4 reverse signaling induces the apoptosis of EBV-transformed B cells through Fas ligand up-regulation. AB - B7-H4 has an inhibitory effect on immune responses via the down-regulation of T cell-mediated immunity, but how the engagement of B7-H4 molecules by counter molecules affects the signaling mechanism of the B7-H4-expressing cells is poorly defined. In this study, we found that B7-H4 expression was enhanced on B cells infected with Epstein-Barr virus (EBV) and that triggering of these molecules induced apoptosis of EBV-transformed B cells. Engagement of B7-H4 initially increased intracellular level of ROS, which then induced the expression of FasL. Engagement of B7-H4 subsequently provoked Fas-mediated and caspase-dependent apoptosis in association with cytochrome c and AIF, and EndoG was released from the mitochondria on EBV-transformed B cells. These results suggest that B7-H4 may be a potential therapeutic target for EBV involved malignancy diseases. PMID- 18417277 TI - Assessment of metal element concentrations in mussel (M. Galloprovincialis) in Eastern Black Sea, Turkey. AB - The main goal of this work is to determine the effects of pollution of copper, lead and zinc mines on the Eastern Black Sea. Metal and heavy metal concentrations in the Eastern Black Sea mussels were measured using Energy Dispersive X-ray Fluorescence (EDXRF) and Flame Atomic Absorption Spectroscopy (FAAS). The analytical results showed that the tissue of mussel in Eastern Black Sea contains K, Ca, Fe, Cu, Zn, and Sr elements, and the shell of mussel contains Ca, Cu, Sr, and Ba elements. Due to the detection limit of EDXRF, the mussels were analyzed with FAAS for Cr, Mn, Ni, Cd and Pb elements. An ANOVA and Pearson correlation analyses were performed. The results showed although that the mean concentrations of Cu and Zn for the tissue of the mussels were markedly above the permissible levels of the Turkish regulations, Zn concentration is in the limits of the Food and Agriculture Organization (FAO). PMID- 18417278 TI - Study of arsenic(V) adsorption on bone char from aqueous solution. AB - Arsenic is a toxic element and may be found in natural waters as well as in industrial waters. Leaching of arsenic from industrial wastewater into groundwater may cause significant contamination, which requires proper treatment before its use as drinking water. The present study described the removal of As(V) on bone char in batch studies conducted as a function of pH, dosage of adsorbent, and contact time. Kinetics revealed that uptake of As(V) ion by bone char was very rapid in the first 30min and equilibrium time was independent of initial As(V) concentration. And the adsorption process followed a first-order kinetics equation. The arsenic removal was strongly dependent on pH and dosage of adsorbent. Fourier transform infrared spectra of bone char before and after As(V) adsorption demonstrated that Ca-OH functional group plays an important role for As(V) ions removal, and the mechanisms of the removal of As(V) on bone char was complex mechanism where both co-precipitation and ion exchange. The results suggested that bone char can be used effectively for the removal of As(V) ion from aqueous solution. PMID- 18417279 TI - Sorption properties of the activated carbon-zeolite composite prepared from coal fly ash for Ni(2+), Cu(2+), Cd(2+) and Pb(2+). AB - Composite materials of activated carbon and zeolite have been prepared successfully by activating coal fly ash (CFA) by fusion with NaOH at 750 degrees C in N(2) followed by hydrothermal treatments under various conditions. Uptake experiments for Ni(2+), Cu(2+), Cd(2+) and Pb(2+) were performed with the materials thus obtained from CFA. Of the various composite materials, that were obtained by hydrothermal treatment with NaOH solution (ca. 4M) at 80 degrees C (a composite of activated carbon and zeolite X/faujasite) proved to be the most suitable for the uptake of toxic metal ions. The relative selectivity of the present sorbents for the various ions was Pb(2+)>Cu(2+)>Cd(2+)>Ni(2+), with equilibrium uptake capacities of 2.65, 1.72, 1.44 and 1.20mmol/g, respectively. The sorption isotherm was a good fit to the Langmuir isotherm and the sorption is thought to progress mainly by ion exchange with Na(+). The overall reaction is pseudo-second order with rate constants of 0.14, 0.17, 0.21 and 0.20Lg/mmol min for the uptake of Pb(2+), Cu(2+), Cd(2+) and Ni(2+), respectively. PMID- 18417280 TI - Ion exchange of ammonium in natural and synthesized zeolites. AB - In this study, zeolite Na-P and Na-Y was prepared by hydrothermal treatment of the Chinese natural clinoptilolite with NaOH. The ion exchange of NH4+ into the three zeolites in the temperature range of 288-333K was also investigated, and the thermodynamic parameters were calculated. The selectivity sequence for NH4+ entering the sodium form of the three materials was Na-clinoptilolite>Na-Y>Na-P, as indicated by values of DeltaG degrees . The results demonstrated that the Si/Al molar ratio of zeolites determined the selectivity for NH4+. PMID- 18417281 TI - Removal of trivalent chromium from water using low-cost natural diatomite. AB - Trivalent chromium was removed from the artificial wastewater using low-cost diatomite in batch and continuous systems. In batch system, four different sizes and five different amount of sorbent were used. The effect of the temperature on sorption was evaluated with using three different temperatures. As a result of the experiments, 85% of the trivalent chromium was removed from the wastewater in conditions of using 1.29mm grain material at 30 degrees C temperature for 60min in batch system but chromium removal was 82% at 30 degrees C temperature for 22min and 97% from the wastewater at 30 degrees C temperature for 80min in continuous system. Also, the equilibrium adsorption isotherms have been analyzed by Langmuir and Freundlich models. The Langmuir isotherms have the highest correlation coefficients. Langmuir adsorption isotherm constants corresponding to adsorption capacity, q0, were found to be 28.1, 26.5 and 21.8mg Cr3+/g diatomite at 15, 30 and 45 degrees C, respectively. Adsorption process was an exothermic process as a result of thermodynamic parameters calculations. The kinetic data of the sorption showed that the pseudo second-order equation was the more appropriate, which indicate that the intraparticle diffusion is the rate-limiting factor. PMID- 18417282 TI - Precipitation of cetyl (hexadecyl) pyridineum chloride using mono and divalent oxyanions. AB - Experimental investigations have been carried out to observe the performance of precipitation behavior of potassium permanganate (KMnO4) and potassium dichromate (K2Cr2O7) in cetyl (hexadecyl) pyridineum chloride (CPC) solution. As in the case of Al3+-dodecylbenzenesulfonate systems [P. Somasundaran, K.P. Anathapadmanabhan, M.S. Celik, Langmuir 4 (1988) 1061-1063], the precipitation is found to be caused by interaction of CPC micelles with oxyanions. The counter oxyanions have a strong tendency to bind themselves to the surface of cationic CPC micelles. This lowered the free oxyanion concentration in solution. Therefore, to start precipitation, higher oxyanion concentration is required for higher CPC concentration. The effects of temperature, concentrations of both counter ions and CPC on the precipitation have been studied in detail. It has been observed that at CPC to KMnO4 concentration ratio of 1.0 (concentration of CPC and KMnO4 is 400ppm), the percentage precipitation of CPC is around 99.3 at 30 degrees C. The percent precipitation of CPC decreases to about 94% when temperature increases to 70 degrees C at the same condition. The extent of CPC precipitation increases at the same experimental condition when K2Cr2O7 is used instead of KMnO4. PMID- 18417283 TI - Co-metabolic degradation of bensulfuron-methyl in laboratory conditions. AB - The present study deals with the degradation of bensulfuron-methyl by microorganisms cultured with different sources of carbon, nitrogen and phosphorus. Addition of carbon source accelerated the degradation of bensulfuron methyl under co-metabolism process. Sodium lactate was the best carbon source for the degradation of bensulfuron-methyl, compared to other carbon sources studied, and the degradation ratio of bensulfuron-methyl reached 79.5%, whereas only 34.6 and 29.7% were removed in the presence of glucose and sucrose, respectively. Supplement of nitrogen source also enhanced degradation of bensulfuron-methyl. However, no significant differences were observed in the loss of bensulfuron methyl between organic nitrogen and inorganic source. Phosphate buffer was supplemented into the media to maintain neutral conditions for the advantage of the strain growth since increase in pH value was observed. An orthogonal array design was applied to arrange main factors singled out for investigating the influence of factor and interaction between them on the degradation of bensulfuron-methyl. Statistical analysis showed that the concentration of sodium lactate, bensulfuron-methyl and inoculum size were the main effects, and the interaction of sodium lactate and bensulfuron-methyl was of statistical significance. PMID- 18417284 TI - Cellular localization of cadmium and structural changes in maize plants grown on a cadmium contaminated soil with and without liming. AB - The effects of different concentrations of soil cadmium (0, 1, 3, 5, 10, and 20mgkg(-1)) on growth, structural changes and cadmium cellular localization in leaves of maize plants (Zea mays L.) were investigated in a pot experiment. The results showed that the structural changes observed in maize leaves were not only a response to the Cd-induced stress but also a cellular mechanism to reduce the free Cd(+2) in the cytoplasm. However, this mechanism seems to be efficient only up to a Cd concentration in leaves between 27 and 35mgkg(-1) for soils without and with liming, respectively. The cellular response varied with both the Cd concentration in soil and liming. For limed soil, Cd was preferentially accumulated in the apoplast while for unlimed soils Cd was more evenly distributed into the cells. The ability of Cd accumulation depended on the leaf tissue considered. The apoplast collenchyma presented the highest Cd concentration followed by the endodermis, perycicle, xylem, and epidermis. On the other hand, symplast Cd accumulated mainly in the endodermis, bundle sheath cells, parenchyma, and phloem. Based on the structural changes and growth reduction, the critical toxic concentration of soil Cd to maize plants is between 5 and 10mgkg(-1). PMID- 18417285 TI - Extractive removal of chromium (VI) from industrial waste solution. AB - Extractive removal of Cr (VI) was carried out from chloride solutions using cyanex 923 mixed with kerosene. The efficiency of this extractant was studied under various experimental conditions, such as concentration of different mineral acids in the aqueous phase, concentration of cyanex 923 and Cr (VI) present in the initial aqueous feed, temperature and time of extraction, organic to aqueous (O/A) phase ratio. Percentage Cr (VI) extraction decreases with the increase in temperature at varying concentration of cyanex 923. The interference of the impurities usually associated with Cr (VI) such as Cr (III), Cu, Ni, Fe (II), Zn, Chloride and sulphate, etc., were examined under the optimized conditions and only Zn was found to interfere. Under the optimum experimental conditions 98.6 99.9% of Cr (VI) was extracted in 3-5 min at O/A of 2 with the initial feed concentration of 1g/L of Cr (VI). The extracted Cr (VI) was quantitatively stripped with 1M NaOH and the organic phase obtained after the stripping of Cr (VI) was washed with dilute HCl solution to neutralize any NaOH trapped/adhered to the solvent and then with distilled water. This regenerated solvent was reused in succeeding extraction of chromium (VI). Finally a few experiments were performed with the synthetic effluent from an electroplating industry. PMID- 18417286 TI - The effect of operational parameters on electrocoagulation-flotation process followed by photocatalysis applied to the decontamination of water effluents from cellulose and paper factories. AB - Cellulose and paper pulp factories utilize a large amount of water generating several undesirable contaminants. The present work is a preliminary investigation that associates the electrocoagulation-flotation (EC) method followed by photocatalysis to treat such wastewater. For EC, the experiment with aluminium and iron electrodes showed similar efficiency. Iron electrodes (anode and cathode) were chosen. By applying 30min of EC/Fe(0), 153A m(-2) and pH 6.0, the COD values, UV-vis absorbance and turbidity underwent an intense decrease. For the subsequent UV photocatalysis (mercury lamps) TiO(2) was employed and the favourable operational conditions found were 0.25g L(-1) of the catalyst and solution pH 3.0. The addition of hydrogen peroxide (50mmol L(-1)) highly increased the photo-process performance. By employing the UV/TiO(2)/H(2)O(2) system, the COD reduction was 88% compared to pre-treated effluents and complete sample photobleaching was verified. The salt concentration on EC (iron electrodes) showed that the electrolysis duration can be reduced from 30 to 10min by the addition of 5.0g L(-1) of NaCl. The biodegradability index (BOD/COD) increased from 0.15 (pre-treated) to 0.48 (after EC) and to 0.89 (after EC/photocatalysis irradiated for 6h), showing that the employed sequence is very helpful to improve the water quality. This result was confirmed by biotoxicity tests performed with microcrustaceous Artemia salina. PMID- 18417287 TI - Removal of total phenols from olive-mill wastewater using an agricultural by product, olive pomace. AB - The purpose of this study was to investigate the sorption of total phenols, which are contained in olive-mill wastewater (OMWW), on solid by-products of olive pomace processing mills. Preliminary batch experiments were conducted using three different types of olive pomace, dried olive pomace (OP-1), dried and solvent extracted olive pomace (OP-2) and dried, solvent extracted and incompletely combusted olive pomace (OP-3). According to the results, OP-3 showed high performance for total phenols sorption and stability. For sorbent concentration of 10gL(-1) and sorbate concentration of 50mgL(-1), more than 40% of initial total phenols concentration was removed. Sorption kinetics was well described by the pseudo-second order rate model (R2>0.99). Total phenols removal efficiency was improved by increasing sorbent concentration and solution's pH or decreasing particle size of the sorbent material. The Langmuir and Freundlich isotherms sufficiently described OP-3 sorption capacity for the concentration range studied (50-500mgL(-1)). Fixed bed sorption experiments showed that lower flow rates and smaller particle size of sorbent resulted in longer column exhaustion time and higher initial removal efficiency. Experiments with thermally or chemically regenerated OP-3 showed that sorption capacity deteriorated after regeneration. PMID- 18417288 TI - The electrochemical investigation of salts partition with ion exchange membranes. AB - The regenaration of acid and base from the solutions containing metallic salts was achieved by salt splitting method (SSM) using not only anion-exchange membranes (AEM) but also cation-exchange membrane (CEM). In these experiments, while the ion exchange membrane was anion-exchange membrane, acid solutions were used as an anolyte and different salts of potassium were used as a catholyte. In addition to these experiments, while the ion exchange membrane was cation exchange membrane, base solutions were used as a catholyte and different salts of potassium were used as an anolyte. The effects of current density, initial concentrations of anolyte and catholyte solutions, the type of salt solution and the type of the ion-exchange membranes on the recovery ratio of bases and acids were investigated. The results of the experiments were investigated with the Statistical Package for Social Sciences (SPSS) program. The obtained results show that this technique can be used not only for recovering the acid and base wastes of industry but also for removing the impurities in order to obtain pure acids and bases in laboratory conditions. PMID- 18417289 TI - AMPA-induced dark cell degeneration is associated with activation of caspases in pyramidal neurons of the rat hippocampus. AB - Various neurons in the central nervous system (CNS) exhibit selective vulnerability to AMPA-induced delayed neurotoxicity known as dark cell degeneration. Hippocampal pyramidal neurons in the CA1 and CA3 regions display such vulnerability that encompasses morphological changes including cytoplasmic and nuclear condensation, neuronal shrinkage, formation of cytoplasmic vacuoles, and general failure of physiology. The present study was undertaken to ascertain the potential involvement of initiator (caspase-9) and executor (caspase-3) caspases in AMPA-receptor-induced dark cell degeneration in pyramidal neurons. Immunohistochemical analyses revealed that immunoreactivity of the active form of caspase-9 and -3 was increased in pyramidal neurons in CA1 and CA3 regions of the hippocampus following AMPA (100 microM). Elevated levels of active caspase-9 immunoreactivity generally preceded elevations in active caspase-3 immunoreactivity. The pan caspase inhibitor FK011 effectively attenuated AMPA induced dark cell degeneration in both CA1 and CA3 regions. Collectively, the data suggest a role for these caspases in mediating AMPA-induced toxicity in pyramidal neurons of the rat hippocampus. PMID- 18417291 TI - Assessing fear in patients with cervical pain: development and validation of the Pictorial Fear of Activity Scale-Cervical (PFActS-C). AB - The fear avoidance model (FAM) postulates that fear of pain or reinjury is a risk factor for persistent pain and disability, because it leads to the avoidance of physical activity. Research on the FAM has not yielded consistent results, which may be attributed to the model itself, but could also be a product of the way fear of movement is assessed. Studies of the FAM have measured fear using verbal scales consisting of items that are often vague and have only an indirect relationship with fear. This study reports on the development of a Pictorial Fear of Activity Scale-Cervical (PFActS-C). The instrument consists of a set of photographs depicting movements in which four factors that determine biomechanical demands on the neck are systematically varied--Direction of Movement, Arm Position, Weight Bearing, and Extremity of Movement. Patients (n=355) who had been involved in motor vehicle collisions with minimal symptoms (n=143) and moderate to severe symptoms (n=212) rated their fear of engaging in a set of activities depicted in the PFActS-C. Based on a principle components analysis, a 19 item measure was developed. Internal consistency (alpha=.98), stability over time (n=44, IntraClass Correlation=.72), and construct validity were all good to excellent. The results indicate that the PFActS-C may be a useful tool for assessing fear of movement in patients with cervical pain. Research is needed to confirm the factor structure of the PFActS-C and to assess the generalizability of the results to other samples with neck pain. PMID- 18417292 TI - A patient-based national survey on postoperative pain management in France reveals significant achievements and persistent challenges. AB - We carried out a national survey on postoperative pain (POP) management in a representative sample (public/private, teaching/non-teaching, size) of 76 surgical centers in France. Based on medical records and questionnaires, we evaluated adult patients 24h after surgery, concerning information: pre and postoperative pain, evaluation, treatment and side effects. A local consultant provided information about POP management. Data were recorded for 1900 adult patients, 69.3% of whom remembered information on POP. Information was mainly delivered orally (90.3%) and rarely noted on the patient's chart (18.2%). Written evaluations of POP were frequent on the ward (93.7%) with appropriate intervals (4.1 (4.0)h), but not frequently prescribed (32.7%). Pain evaluations were based on visual analog scale (21.1%), numerical scale (41.2%), verbal scale (13.8%) or non-numerical tool (24%). Pain was rarely a criterion for recovery room discharge (19.8%). Reported POP was mild at rest (2.7 (1.3)), moderate during movement (4.9 (1.9)) and intense at its maximal level (6.4 (2.0)). Incidence of side effects was similar according to patient (26.4%) or medical chart (25.1%) including mostly nausea and vomiting (83.3%). Analgesia was frequently initiated during anesthesia (63.6%). Patient-controlled analgesia (21.4%) was used less frequently than subcutaneous morphine (35.1%) whose prescription frequently did not follow guidelines. Non-opioid analgesics used included paracetamol (90.3%), ketoprofen (48.5%) and nefopam (21.4%). Epidural (1.5%) and peripheral (4.7%) nerve blocks were under used. Evaluation (63.4%) or treatment (74.1%) protocols were not available for all patients. This national, prospective, patient-based, survey reveals both progress and persistent challenges in POP management. PMID- 18417293 TI - The value of plant extracts with antioxidant activity in attenuating coccidiosis in broiler chickens. AB - Coccidiosis remains one of the most important diseases in the poultry industry and results in the annual loss of millions of US dollars by the poultry industry. In South Africa and other developing countries where a large percentage of the population is unemployed, cheap food production is necessary. If the control of the coccidian parasite could be made more economical, these savings could be passed on to the consumer. In Europe, where the economics are different, people are becoming more aware of the potential dangers of using antimicrobials in producing animal protein. A solution to both these problems could be the use of plant products that function by mechanisms other than those of chemotherapeutics, with the additional advantage of a natural origin. Antioxidant compounds could hold promise for the control of Eimeria infections due to the association of coccidial infection with lipid peroxidation of the intestinal mucosa. Four plant extracts with antioxidant activity were screened for their anticoccidial activity in vivo with toltrazuril as the positive control. Combretum woodii (160 mg/kg) proved to be extremely toxic to the birds, while treatment with Tulbaghia violacea (35 g/kg), Vitis vinifera (75 mg/kg) and Artemisia afra (150 mg/kg) resulted in feed conversion ratios similar to toltrazuril, and higher than the untreated control. T. violacea also significantly decreased the oocyst production in the birds. From this study we conclude that antioxidant-rich plant extracts have potential benefits in treating coccidial infections. The promising results obtained with T. violacea justify further studies on the potential value of the plant as a therapeutic or prophylactic anticoccidial agent. PMID- 18417294 TI - Attenuation of heart rate control and neural degeneration in nucleus ambiguus following chronic intermittent hypoxia in young adult Fischer 344 rats. AB - Chronic intermittent hypoxia (CIH) attenuates baroreflex control of heart rate (HR). In this study, we assessed whether CIH exposure reduced nucleus ambiguus (NA) control of HR and induced neural degeneration in the NA. Fischer 344 (age: 3 4 months) rats were exposed to either room air (RA: normoxia) or intermittent hypoxia for 35-50 days. At the end of these exposures, animals were anesthetized with pentobarbital. HR responses to arterial blood pressure (AP) changes induced by phenylephrine (PE) and sodium nitroprusside (SNP) were measured. In another set of rats, HR and AP responses to L-glutamate (L-Glu) microinjections (10 mM, 20 nl) into the left NA and electrical stimulation of the left cervical vagus nerve at 1-30 Hz (0.5 mA, 1 ms) for 20 s were measured. Brainstem slices at the level of -800, -400, 0, +400, +800 microm relative to the obex were processed in additional rats using Nissl staining. The NA was identified by retrogradely labeling vagal motoneurons using the tracer tetramethylrhodamine dextran (TMR-D) which was injected into the ipsilateral nodose ganglion. We found that CIH significantly 1) reduced the baroreflex control of HR (slope RA: -1.2+/-0.2 bpm/mmHg; CIH -0.5+/-0.1 bpm/mmHg; P<0.05); 2) attenuated the HR responses to l Glu injections into the NA [HR: -280+/-15 (RA) vs. -235+/-16 (CIH) beats/min; P<0.05]; 3) augmented the HR responses to electrical stimulation of the vagus (P<0.05); 4) induced a significant cellular loss in the NA region (P<0.05). Thus, CIH induces a cell loss in the NA region which may contribute to attenuation of baroreflex sensitivity and NA control of HR following CIH. PMID- 18417295 TI - Family functioning of out-of-treatment cocaine base paste and cocaine hydrochloride users. AB - Knowledge of family structure and behavioral dynamics of out-of-treatment drug users under poverty becomes relevant, because of the role that the family plays in drug use and rehabilitation. HYPOTHESES: 1. The perception of drug users about their family functioning reveals a dysfunctional organization and communication connection problems with their families; and 2. the family system of cocaine base paste (CBP) users presents greater dysfunctionality than cocaine hydrochloride (CH) users. METHOD: Cross-sectional descriptive design of primary users of CH (n=236) and primary users of CBP (n=231) during the last month, out-of-substance abuse treatment during the last 6 months. INSTRUMENTS: Risk Behavior Questionnaire and How-Is-Your-Family Questionnaire. RESULTS: The total sample presented 72.9% of families with risk functioning; CBP users registered a more deteriorated family structure and communication-connection than CH users. CONCLUSIONS: CBP and CH users, who are hidden from health treatment services, do indeed present a high degree of family dysfunction; and the CBP group, compared to the CH group, presented various indicators of greater risk in their family dysfunction. PMID- 18417296 TI - Defect in lipid rafts results in failed tolerance induction at the maternal-fetal interface: a possible cause for the recurrent spontaneous abortion. AB - The pregnancy is the successful induction and maintenance of maternal tolerance to semi-allogeneic fetus. As a failure result, recurrent spontaneous abortion occurs in about 1-2% of women of reproductive age, defined as the loss of three or more consecutive pregnancies. The mechanism of recurrent spontaneous abortion is often elusive. Recently, mounting evidence suggests that HLA-G induced suppressive uterine natural killer (uNK) cells play an important role in the maternal-fetal tolerance and their abnormalities are responsible for recurrent spontaneous abortion. However, there are some clinical cases of recurrent spontaneous abortion could not be detected of the HLA-G alterations, while their uNK cells showed considerable cytolytic activities against fetus. Thus we hypothesize that lipid rafts, specialized micro-domains in plasma membrane, is of vital importance in the HLA-G-NK cells interactions and the accompanied suppressive induction process. We further hypothesize that the defect in lipid rafts may result in failed tolerance induction at the maternal-fetal interface, which would be a novel explanation for the occurrence of recurrent spontaneous abortion. The hypothesis can be practically evaluated by in vitro experiments and clinical tests. To sum up, this hypothesis proposes a new mechanism in the NK cells suppressive induction. Also the hypothesis may provide new vision in the drug development and disease control of recurrent spontaneous abortion. PMID- 18417297 TI - Poly(ADP-ribose) signal in seizures-induced neuron death. AB - Poly(ADP-ribose) is found to be involved in many physiological or pathological processes. It is mainly modulated by poly(ADP-ribose) polymerase (PARP) and poly(ADP-ribose) glycohydrolase (PARG). Either PARP or PARG is associated with the neuronal death in a variety of neurodegenerative diseases. Cumulative data have suggested that poly(ADP-ribose) regulation might have a therapeutic value in neurotoxicity-induced neuron damage, probably due to the inhibition of apoptosis, suppressing of inflammation and activation of cell survival signaling. We hypothesize poly(ADP-ribose) play an important role in seizures-induced neuron death. Seizures can lead to neuron degeneration as for the exitotoxity of glutamate. Recently, it is indicated seizures also can trigger PARP activation. Further investigation is needed to determine whether poly(ADP-ribose) signal is a therapeutic target for seizures-induced injury. PMID- 18417298 TI - Asymmetric testicular levels in the crotch: a thermodynamic perspective. PMID- 18417299 TI - Prospective evaluation to establish a dose response for clinical oral mucositis in patients undergoing head-and-neck conformal radiotherapy. AB - PURPOSE: We conducted a clinical study to correlate oral cavity dose with clinical mucositis, perform in vivo dosimetry, and determine the feasibility of obtaining buccal mucosal cell samples in patients undergoing head-and-neck radiation therapy. The main objective is to establish a quantitative dose response for clinical oral mucositis. METHODS AND MATERIALS: Twelve patients undergoing radiation therapy for head-and-neck cancer were prospectively studied. Four points were chosen in separate quadrants of the oral cavity. Calculated dose distributions were generated by using AcQPlan and Eclipse treatment planning systems. MOSFET dosimeters were used to measure dose at each sampled point. Each patient underwent buccal sampling for future RNA analysis before and after the first radiation treatment at the four selected points. Clinical and functional mucositis were assessed weekly according to National Cancer Institute Common Toxicity Criteria, Version 3. RESULTS: Maximum and average doses for sampled sites ranged from 7.4-62.3 and 3.0-54.3 Gy, respectively. A cumulative point dose of 39.1 Gy resulted in mucositis for 3 weeks or longer. Mild severity (Grade 75% of days) presented a lower incidence of diarrhea (6.5% versus 20.3%, P = 0.02; and 5.9% versus 19.8%, P = 0.05, respectively). CONCLUSION: Providing enteral nutrition to the hospitalized elderly during the summer months is associated with a higher risk of diarrhea. Strategies aimed toward improvement in the quality of enteral nutrition practices should be evaluated to minimize this deleterious clinical outcome. PMID- 18417322 TI - Regional gray matter abnormalities in panic disorder: a voxel-based morphometry study. AB - Although abnormalities in brain structures involved in the neurobiology of fear and anxiety have been implicated in the pathophysiology of panic disorder (PD), relatively few studies have made use of voxel-based morphometry (VBM) magnetic resonance imaging (MRI) to determine structural brain abnormalities in PD. We have assessed gray matter volume in 19 PD patients and 20 healthy volunteers using VBM. Images were acquired using a 1.5 T MRI scanner, and were spatially normalized and segmented using optimized VBM. Statistical comparisons were performed using the general linear model. A relative increase in gray matter volume was found in the left insula of PD patients compared with controls. Additional structures showing differential increases were the left superior temporal gyrus, the midbrain, and the pons. A relative gray matter deficit was found in the right anterior cingulate cortex. The insula and anterior cingulate abnormalities may be relevant to the pathophysiology of PD, since these structures participate in the evaluation process that ascribes negative emotional meaning to potentially distressing cognitive and interoceptive sensory information. The abnormal brain stem structures may be involved in the generation of panic attacks. PMID- 18417323 TI - Preparation and characterization of novel polymeric micelles for 9-nitro-20(S) camptothecin delivery. AB - 9-Nitro-20(S)-camptothecin (9-NC) has achieved remarkable curative effect in anticancer research. However, the clinical application of 9-NC is largely hampered by its poor solubility and stability. In this paper, novel amphiphilic block copolymers derived from d,l-lactide, trimethylene carbonate, and methylated poly(ethylene glycol) (mPEG) (PECA) with different molecular weight were synthesized and characterized. Self-assembly PECA micelles loaded with 9-NC were prepared. The micelles were regular spheres with a diameter ranged from 20 to 120 nm. The critical micelle concentration (CMC) decreased with the increase of the hydrophobic components. The solubility of 9-NC was improved obviously with micelle encapsulation. The stability experiments proved that over 90% of 9-NC could keep its lactone form in micelle solution after incubating in phosphate buffered saline for 100 min, while the corresponding proportion for free drug solution was 25%. The release of 9-NC was nearly zero-order after the burst release, and the long hydrophobic chain length led to slower release rate. The novel PECA copolymer micelles could be effective carriers to improve the solubility, stability, and release performance of 9-NC. PMID- 18417324 TI - [Torticollis in children]. PMID- 18417325 TI - A new choanoflagellate species from Taiwan: morphological and molecular biological studies of Diplotheca elongata nov. spec. and D. costata. AB - A new species of acanthoecid choanoflagellate isolated from brackish waters of the Danshui estuary in North Taiwan has a mineralized lorica that consists of two chambers with a total length of 19-36microm. It shares with Diplotheca costata the features of a posterior lorica chamber formed from broad and flattened costal strips and an anterior chamber with spatula-shaped costal strips. The new species has therefore been placed in the same genus and named Diplotheca elongata. A phylogenetic analysis of partial SSU rDNA sequences from Diplotheca costata and D. elongata supports this taxonomic affiliation. This is a large and distinctive choanoflagellate which has not been reported in any previous study, suggesting that it may be an endemic species of restricted distribution. PMID- 18417326 TI - Morphology and morphogenesis of a new marine ciliate, Apokeronopsis bergeri nov. spec. (Ciliophora, Hypotrichida), from the Yellow Sea, China. AB - The morphology and morphogenesis of a new marine hypotrich ciliate, Apokeronopsis bergeri nov. spec., collected from mussel-farming waters near Qingdao, China, are described from living and protargol-impregnated specimens. This ciliate has characteristics that place it in the family Pseudokeronopsidae, namely, two long rows of frontal cirri (bicorona), which are continuous with the long midventral rows, and a single row of marginal cirri on each side of the body. It shares with its only congener, Apokeronopsis crassa, the long rows of buccal and transverse cirri and the wide spacing between the midventral rows of cirri. These characters separate the genus Apokeronopsis from Pseudokeronopsis, which has a single buccal cirrus, fewer transverse cirri and midventral rows of cirri arranged in a typical zig-zag pattern. A. bergeri differs from A. crassa in its shape, colour and in the numbers of membranelles and transverse cirri. Although morphogenesis in A. bergeri is similar to that of A. crassa in most respects, the mode of formation of the buccal cirri is slightly different. The close relationship of A. bergeri with A. crassa, and the more distant relationship with three Pseudokeronopsis species, is supported by a comparison of the sequences of their ITS1-5.8S-ITS2 rDNA regions. PMID- 18417327 TI - Autonomous virtual mobile robot for three-dimensional medical image exploration: application to micro-CT cochlear images. AB - OBJECTIVE: In this paper, we present an autonomous virtual mobile robot (AVMR) for three-dimensional (3D) exploration of unknown tubular-like structures in 3D images. METHODS AND MATERIALS: The trajectory planning for 3D central navigation is achieved by combining two neuro-fuzzy controllers, and is based on 3D sensory information; a Hough transform is used to locally fit a cylinder during the exploration, estimating the local radius of the tube. Nonholonomic constraints are applied to assure a smooth, continuous and unique final path. When applied to 3D medical images, the AVMR operates as a virtual endoscope, directly providing anatomical measurements of the organ. After a thorough validation on challenging synthetic environments, we applied our method to eight micro-CT datasets of cochleae. RESULTS: Validation on synthetic environments proved the robustness of our method, and highlighted key parameters for the design of the AVMR. When applied to the micro-CT datasets, the AVMR automatically estimated length and radius of the cochleae: results were compared to manual delineations, proving the accuracy of our approach. CONCLUSIONS: The AVMR presents several advantages when used as a virtual endoscope: the nonholonomic constraint guarantees a unique and smooth central path, which can be reliably used both for qualitative and quantitative investigation of 3D medical datasets. Results on the micro-CT cochleae are a significant step towards the validation of more clinical computed tomography (CT) studies. PMID- 18417329 TI - Bioprocess of uniform-sized crosslinked chitosan microspheres in rats following oral administration. AB - Chitosan microspheres have a great potential in pharmaceutical application. In this study, uniform-sized chitosan microspheres crosslinked with glutaraldehyde (CG microspheres) were prepared by Shirasu Porous Glass (SPG) membrane emulsification technique. Based on the characterizations of uniform size and autofluorescence, it was possible to develop a new detecting system for observing and quantifying the CG microspheres in rats with three different diameters (2.1, 7.2 and 12.5 microm) synchronously after oral administration. This system was a combination of scanning electron microscopy (SEM), laser scanning confocal microscope (LSCM) and flow cytometer technique, which showed the advantages of being simple, intuitionistic, repeatable and sensitive. After oral administration of three kinds of particles with different diameters, bioadhesion in gastrointestinal tract, absorption in gastrointestinal tract, distribution in systemic tissues, and biodegradation in reticuloendothelial system (RES) were studied firstly in detail. The CG microspheres showed different fates in bioadhesion, absorption and distribution according to their diameters, while the biodegradation also varied due to the different locations in RES. These original results would indicate a better way for the CG microspheres in the clinical application. PMID- 18417328 TI - Modulation of gel formation and drug-release characteristics of lidocaine-loaded poly(vinyl alcohol)-tetraborate hydrogel systems using scavenger polyol sugars. AB - Polyol sugars, displaying a plurality of hydroxyl groups, were shown to modulate tetrahydroxyborate (borate) cross-linking in lidocaine hydrochloride containing poly(vinyl alcohol) semi-solid hydrogels. Without polyol, demixing of borate cross-linked PVA hydrogels into two distinct phases was noticeable upon lidocaine hydrochloride addition, preventing further use as a topical system. d-Mannitol incorporation was found to be particularly suitable in circumventing network constriction induced by ionic and pH effects upon adding the hydrochloride salt of lidocaine. A test formulation (4% w/v lidocaine HCl, 2% w/v d-mannitol, 10% w/v PVA and 2.5% w/v THB) was shown to constitute an effective delivery system, which was characterised by an initial burst release and a drug release mechanism dependent on temperature, changing from a diffusion-controlled system to one with the properties of a reservoir system. The novel flow properties and innocuous adhesion of PVA-tetrahydroxyborate hydrogels support their application for drug delivery to exposed epithelial surfaces, such as lacerated wounds. Furthermore, addition of a polyol, such as d-mannitol, allows incorporation of soluble salt forms of active therapeutic agents by modulation of cross-linking density. PMID- 18417330 TI - Development of microporous drug-releasing films cast from artificial nanosized latexes of poly(styrene-co-methyl methacrylate) or poly(styrene-co-ethyl methacrylate). AB - Two sets of copolymers comprising of styrene and either methyl or ethyl methacrylate as comonomer were conveniently synthesized by microemulsion copolymerization. The purified materials were characterized by GPC-MALLS and were shown to form artificial nanolatexes in THF. ATR-FTIR analysis revealed differences in copolymer composition and based on the copolymer properties, a selection of copolymers was chosen to cast drug-loaded, microporous films that exhibit microencapsulation of drug agglomerates. The contact angles of the copolymers suggested potential applications in medical devices to prevent the formation of bacterial biofilms that commonly result in infections. Additionally, the different copolymeric films showed two phases of drug release characterized by a rapid initial drug release followed by a zero-order phase. Depending on the application, one could select the copolymer films that best suited the application i.e. for short-term drug release applications such as urinary catheters or long-term applications such as artificial implants. PMID- 18417331 TI - Sustained release of bioactive glycosylated glial cell-line derived neurotrophic factor from biodegradable polymeric microspheres. AB - Glial cell-line derived neurotrophic factor (GDNF), a potent neurotrophic factor for dopaminergic neurons, appeared as a promising candidate for treating Parkinson's disease. GDNF microencapsulation could ensure protection against degradation due to the fragile nature of the protein. Poly(lactide-co-glycolide) (PLGA) microparticles loaded with recombinant glycosylated GDNF obtained in a mammalian cell line were prepared by TROMS, a semi-industrial technique capable of encapsulating fragile molecules maintaining their native properties. The effects of several parameters as PLGA copolymer type, PEG 400 quantity co encapsulated with GDNF or drug loading, on the properties of the particles were investigated. Microparticles showed a mean diameter between 8 and 30 microm, compatible with their stereotaxic implantation. The drug entrapment efficiency ranged from 50.6% to 100% depending on the microsphere composition. GDNF was better encapsulated using hydrophilic polymers with high molecular weight such as RG 503H. In vitro drug release was influenced by the polymer type as well as by the amount of PEG 400 co-encapsulated with GDNF. Microparticles prepared using PLGA RG 503H released 67% of the total protein content within 40 days. Moreover, very low concentrations of poly(vinyl alcohol) were detected after microparticles washing and freeze-drying. Finally, a PC-12 bioassay demonstrated that the in vitro GDNF released was bioactive. PMID- 18417332 TI - Wnt signaling in somite development. AB - During vertebrate embryogenesis, specialized mesodermal structures, called somites, give rise to a variety of mesodermal tissues including skeletal muscles, vertebrae and dermis. Development of the somites is a rhythmic process that involves a series of steps including segmentation of the paraxial mesoderm, epithelialization, somite formation, somite maturation, somite patterning and differentiation of somitic cells into different lineages. Wnt signaling has been found to play crucial roles in multiple steps of somite development. In this review, we present a brief overview of current knowledge on Wnt signaling events during the development of somites and their derivatives. PMID- 18417333 TI - Clinical anatomy of the calcaneal tuberosity. AB - The aim of the study was a qualitative anatomical analysis of the macroscopic features of the surface of the calcaneal tuberosity, of the architecture of its cancellous bone and histological structure of the whole region. Dry human bones and pathological dissection material 24-36 h post mortem were used in the study. On the tuberosity, the variability of its surface relief and the two borders between the superior, middle and inferior facets were studied. More frequent medial declination of the inferior line, corresponding to the distal circumference of the Achilles tendon attachment, was found. Two systems of expressive condensation of cancellous bone just below the surface of the calcaneal tuberosity were described. In the histological part of the study, the distribution and different thickness of the fibrous cartilage layer covering the attachment region of Achilles tendon, the bottom of retro-calcaneal bursa and the whole surface of the calcaneal tuberosity were described. The functional and clinical relevance of results obtained are evaluated from the point of view of disciplines dealing with the pathology and surgery of the heel region. The relationships of official anatomical terms and a wide spectrum of clinical synonyms designating this region are discussed. PMID- 18417334 TI - An amino acid substitution in PBP-3 in Haemophilus influenzae associate with the invasion to bronchial epithelial cells. AB - Haemophilus influenzae is a common pathogen of respiratory infections. We examined whether beta-lactamase-negative ampicillin-resistant (BLNAR) strains that are known to have ampicillin resistance due to a substitution of amino acid of penicillin binding protein (PBP)-3, differ from beta-lactamase-negative ampicillin-susceptible strains with regard to invasion of bronchial epithelium. After 3h incubation of each of 34 beta-lactamase-negative ampicillin-susceptible and 57 BLNAR strains in the presence of BEAS-2B cells, a human bronchial epithelium cell line, extracellular bacteria were killed using gentamicin and intracellular bacteria numbered. All nine strains in which the efficiency of invasion was 1% or higher were BLNAR strains. The rate of invasion was significantly greater in strains with PBP-3 amino acid substitution (Met377 to Ile, Ser385 to Thr, Leu389 to Phe, and Asn526 to Lys) (n=34) than in those with no amino acid substitution. Electron microscopy showed that high invasive BLNAR strains were observed in cytoplasm of BEAS-2B cell layer. The injured cells were 9.44+/-1.76% among attaching cells examined by trypan blue staining after 6h. These data may suggest that the amino acid substitution of the PBP in BLNAR strains may at least partly play roles in macropinocytosis, leading to the invasion and injury to epithelial cells. PMID- 18417335 TI - Assay to mechanically tune and optically probe fibrillar fibronectin conformations from fully relaxed to breakage. AB - In response to growing needs for quantitative biochemical and cellular assays that address whether the extracellular matrix (ECM) acts as a mechanochemical signal converter to co-regulate cellular mechanotransduction processes, a new assay is presented where plasma fibronectin fibers are manually deposited onto elastic sheets, while force-induced changes in protein conformation are monitored by fluorescence resonance energy transfer (FRET). Fully relaxed assay fibers can be stretched at least 5-6 fold, which involves Fn domain unfolding, before the fibers break. In native fibroblast ECM, this full range of stretch-regulated conformations coexists in every field of view confirming that the assay fibers are physiologically relevant model systems. Since alterations of protein function will directly correlate with their extension in response to force, the FRET vs. strain curves presented herein enable the mapping of fibronectin strain distributions in 2D and 3D cell cultures with high spatial resolution. Finally, cryptic sites for fibronectin's N-terminal 70-kD fragment were found to be exposed at relatively low strain, demonstrating the assay's potential to analyze stretch-regulated protein-protein interactions. PMID- 18417337 TI - Pilot project of biogas production from pig manure and urine mixture at ambient temperature in Ventanilla (Lima, Peru). AB - Parque Porcino de Ventanilla has an extension of 840ha with 2200 farmers dedicated to pig production. There is a lack of services in the area (i.e., water supply, electricity, or waste collection). Anaerobic treatment of pig manure would replace current dumping and incineration, reducing environmental pollution and hazards to public health, as well as providing an organic fertilizer and biogas. The objective of the present work was to study the viability of ambient temperature anaerobic digestion of pig manure diluted in urine, by means of on site pilot scale reactors. The final goal was to establish design parameters for anaerobic digesters to be implemented; since it was part of a project to improve life conditions for the farmers through the incorporation of better management techniques. Experiments were carried out in a low-cost pilot plant, which consists of three anaerobic digesters (225L total volume), without heating or agitation, placed in a greenhouse. The start-up of the digestion process was performed with a mixture of temperature adapted pig manure-sludge and fresh rumen, and showed a good performance regardless of the dilution of pig manure with water or urine, which is a key parameter due to the scarcity of water in the area under study. PMID- 18417336 TI - Cytokine-producing B lymphocytes-key regulators of immunity. AB - The successful use of B cell depletion therapy for the treatment of autoimmune disease has led to a resurgent appreciation of B cells as powerful regulators of immunity. However, to the surprise of many, B cells appear to regulate autoimmune conditions independently of their ability to produce autoantibodies. Indeed, disturbances in the ability of B cell subsets to present antigen, produce cytokines, and regulate the activities of T cells is emerging as a key feature in many inflammatory diseases. Here we review the recent literature describing cytokine-producing regulatory and effector B cell subsets in health and disease and discuss how future B cell-directed therapies might target the pathologic cytokine-producing effector B cell subsets without impacting the protective regulatory subsets. PMID- 18417338 TI - Convergence on chromatin of non-genomic and genomic pathways of hormone signaling. AB - Gene regulation by steroid hormones involves genomic and non-genomic signaling pathways and the relationship between these two pathways is unknown. Genomic actions are often mediated by binding of the ligand-activated hormone receptors to hormone responsive elements (HREs) followed by recruitment of co-regulators, remodeling of chromatin and formation of the transcription initiation complex. The non-genomic effects of steroid hormones involve the rapid and transient activation of several kinase cascades often mediated by a subpopulation of "nuclear" receptors located in the cytoplasmic side of the cell membrane. The progesterone effect on breast cancer cell proliferation involves activation of the Src/Ras/Erk cascade mediated by a specific interaction between two domains of the N-terminal half of PR and the ligand-binding domain of ERalpha. Unexpectedly, selective inhibition of Erk, or its target kinase Msk1, interferes with chromatin remodeling and blocks MMTV transcriptional activation. A complex of activated PR, Erk and Msk1 is recruited to promoter already 5 min after hormone treatment and phosphorylates histone H3 at serine 10, leading to displacement of HP1gamma, as a requisite for recruitment of Src1, chromatin remodeling complexes (hSnf2h and Brg1) and RNA polymerase II. Thus, activation of signaling cascades in the cytoplasm is essential for chromatin remodeling and transcriptional activation of a subset of steroid hormone target genes. PMID- 18417339 TI - Experimental and kinetic modelling studies on the acid-catalysed hydrolysis of the water hyacinth plant to levulinic acid. AB - A comprehensive experimental and modelling study on the acid-catalysed hydrolysis of the water hyacinth plant (Eichhornia crassipes) to optimise the yield of levulinic acid (LA) is reported (T=150-175 degrees CH2SO4 = 0.1-1M, water hyacinth intake=1-5wt%). At high acid concentrations (>0.5M), LA was the major organic acid whereas at low acid concentrations (<0.1M) and high initial intakes of water hyacinth, the formation of propionic acid instead of LA was favoured. The highest yield of LA was 53mol% (35wt%) based on the amount of C6-sugars in the water hyacinth (T=175 degrees CH2SO4 =1M , water hyacinth intake=1wt%). The LA yield as a function of the process conditions was modelled using a kinetic model originally developed for the acid-catalysed hydrolysis of cellulose and good agreement between the experimental and modelled data was obtained. PMID- 18417340 TI - Enhancement of bioremediation by Ralstonia sp. HM-1 in sediment polluted by Cd and Zn. AB - In this study, the potential for the application of the bioaugmentation to Cd and Zn contaminated sediment was investigated. A batch experiment was performed in the lake sediments augmented with Ralstonia sp. HM-1. The degradation capacity of 18.7 mg-DOC/l/day in the treatment group was bigger than that of the blank group (4.4 mg-DOC/l/day). It can be regarded as the result of the reduction of the metal concentration in the liquid phase due to adsorption into the sediments, with the increased alkalinity resulting from the reduction of sulfate by sulfate reducing bacteria (SRB). The removal efficiency of cadmium and zinc in the treatment group was both 99.7% after 35 days. Restrain of elution to water phase from sediment in the Ralstonia sp. HM-1 added treatment group was also shown. In particular, the observed reduction of the exchangeable fraction and an increase in the bound to organics or sulfide fraction in the treatment group indicate its role in the prevention of metal elution from the sediment. Therefore, for bioremediation and restrain of elution from the sediment polluted by metal, Ralstonia sp. augmentation with indigenous microorganism including SRB, sediment stabilization and restrain of elution to surface water is recommended. PMID- 18417341 TI - Cellulosic hydrogen production with a sequencing bacterial hydrolysis and dark fermentation strategy. AB - In this study, cellulose hydrolysis activity of two mixed bacterial consortia (NS and QS) was investigated. Combination of NS culture and BHM medium exhibited better hydrolytic activity under the optimal condition of 35 degrees C, initial pH 7.0, and 100rpm agitation. The NS culture could hydrolyze carboxymethyl cellulose (CMC), rice husk, bagasse and filter paper, among which CMC gave the best hydrolysis performance. The CMC hydrolysis efficiency increased with increasing CMC concentration from 5 to 50g/l. With a CMC concentration of 10g/l, the total reducing sugar (RS) production and the RS producing rate reached 5531.0mg/l and 92.9mg/l/h, respectively. Furthermore, seven H2-producing bacterial isolates (mainly Clostridium species) were used to convert the cellulose hydrolysate into H2 energy. With an initial RS concentration of 0.8g/l, the H2 production and yield was approximately 23.8ml/l and 1.21mmol H2/g RS (0.097mmol H2/g cellulose), respectively. PMID- 18417342 TI - A refined pharmacophore model for HIV-1 integrase inhibitors: Optimization of potency in the 1H-benzylindole series. AB - We report herein the development of a new three-dimensional pharmacophore model for HIV-1 integrase inhibitors which led to the discovery of some 4-[1-(4 fluorobenzyl)-1H-indol-3-yl]-2-hydroxy-4-oxobut-2-enoic acids that are able to specifically inhibit the strand transfer step of integration at nanomolar concentration. The synthesis of the new designed molecules is also described. PMID- 18417343 TI - Pyridyl-phenyl ether monoamine reuptake inhibitors: Impact of lipophilicity on dual SNRI pharmacology and off-target promiscuity. AB - A novel series of pyridyl-phenyl ethers are disclosed, which possess dual 5-HT and NA reuptake pharmacology with good selectivity over dopamine reuptake inhibition. An analysis of the relationship between lipophilicity and pharmacology highlighted that potent dual SNRI activity was only achievable at c log P>3.5. The series was found to possess significant polypharmacology issues, and we concluded that this off-target promiscuity was related to lipophilicity. PMID- 18417344 TI - Identification of acridinyl hydrazides as potent aspartic protease inhibitors. AB - We have identified acridinyl derivatives as potent aspartic protease inhibitors by virtual screening of in-house library of synthetic compounds. Enzyme inhibition experiments showed that both compounds inhibit human cathepsin D and Plasmodium falciparum plasmepsin-II in nanomolar ranges. The IC(50) values against cathepsin D and plasmepsin-II of compound-Nar103 were found to be 9.0+/ 2.0 and 4.0+/-1.0nM and of compound-Nar110 were 0.5+/-0.05 and 0.13+/-0.03nM, respectively. Ligand docking predicted the binding of acridinyl derivatives at the substrate-binding cleft, where hydrazide part of the inhibitors interact with the S1-S1' subsite residues including catalytic aspartates. The phenyl ring and acridinyl moiety of the inhibitors were predicted to interact with S2/S3 and S2'/S3' subsite residues. PMID- 18417345 TI - Normative data for the dynamic pedobarographic profiles of children. AB - In order to establish the clinical utility of pedobarography in the treatment of childhood foot pathology, a reliable set of pedobarograph data describing non pathologic feet is required. The purpose of this study was to describe the pedobarographic profiles of normal children across all ages, with specific focus on young children and explore age-related differences in foot pressure patterns. The Tekscan HR Mat pressure measurement system was used in a protocol involving a dynamic test at self-selected speed and walking pattern of 146 normal children (age range 1.6-14.9 years). Relative force and timing data were obtained across five foot segments (heel, lateral midfoot, medial midfoot, lateral forefoot, and medial forefoot). Analysis of variance (ANOVA) techniques were applied to determine if there were any age-related differences in foot pressure profiles in children across four a priori pedobarograph variables: % of stance at initiation at the heel, % of stance at initiation at the medial midfoot, maximum % force at the heel, maximum % force at the medial midfoot. Differences in foot pressure profiles were distinguished across three age groups: (1) Group 1: <2 years; (2) Group 2: 2-5 years; and (3) Group 3: >5 years. Age-related differences in initiation patterns, force transmission, and the amount of time spent on each foot segment provide evidence for maturation of children's foot pressure profiles from a flatfoot pattern in the young child to a curvilinear pattern in the older child. PMID- 18417346 TI - Pre-operative mental wellbeing and the outcome of knee replacement. AB - This study aimed to determine if a difference exists between the mental well being of patients prior to total knee, medial unicompartmental and patellofemoral joint replacement and then assessed its effect on outcome. One hundred and thirteen patients were identified (41 TKR, 37 UKR and 35 PFJR). All patients had mental health assessed pre-operatively using SF-12. The reduced WOMAC (rWOMAC) score was used pre-operatively and then at 8 and 24 months post-operation to assess outcome. Reduced pre-operative mental wellbeing was found in 54% of the patients. There was no statistically significant difference demonstrated between the mean mental component scores (MCS) between the three operation groups. The mean rWOMAC at 24 months in the MCS<40 group was 28 compared to 17 in those with high mental well being (MCS>60). Pre-operative MCS had a significant effect, such that reduced mental wellbeing was associated with a worse 24 month outcome. PMID- 18417348 TI - Pharmacological and pharmacokinetic characterization of 2-piperazine-alpha isopropyl benzylamine derivatives as melanocortin-4 receptor antagonists. AB - A series of 2-piperazine-alpha-isopropylbenzylamine derivatives were synthesized and characterized as melanocortin-4 receptor (MC4R) antagonists. Attaching an amino acid to benzylamines 7 significantly increased their binding affinity, and the resulting compounds 8-12 bound selectively to MC4R over other melanocortin receptor subtypes and behaved as functional antagonists. These compounds were also studied for their permeability using Caco-2 cell monolayers and metabolic stability in human liver microsomes. Most compounds exhibited low permeability and high efflux ratio possibly due to their high molecular weights. They also showed moderate metabolic stability which might be associated with their moderate to high lipophilicity. Pharmacokinetic properties of these MC4R antagonists, including brain penetration, were studied in mice after oral and intravenous administrations. Two compounds identified to possess high binding affinity and selectivity, 10d and 11d, were studied in a murine cachexia model. After intraperitoneal (ip) administration of 1mg/kg dose, mice treated with 10d had significantly more food intake and weight gain than the control animals, demonstrating efficacy by blocking the MC4 receptor. Similar in vivo effects were also observed when 11d was dosed orally at 20mg/kg. These results provide further evidence that a potent and selective MC4R antagonist has potential in the treatment of cancer cachexia. PMID- 18417347 TI - Redefining the structure-activity relationships of 2,6-methano-3-benzazocines. Part 6: Opioid receptor binding properties of cyclic variants of 8 carboxamidocyclazocine. AB - A series of 7,8- and 8,9-fused pyrimidinone, aminopyrimidine and pyridone derivatives of 8-carboxamidocyclazocine (8-CAC) have been prepared and evaluated in opioid receptor binding assays. Targets were designed to corroborate a pharmacophore hypothesis regarding the bioactive conformation of the carboxamide of 8-CAC. In addition to the results from this study strongly supporting this pharmacophore hypothesis, a number of novel compounds with high affinity to opioid receptors have been identified. PMID- 18417349 TI - Distribution and uptake of 137Cs in relation to alkali metals in a perhumid montane forest ecosystem. AB - We determined the content of radiocesium ((137)Cs) and alkali metals in soils, plants (2 ferns, a shrub and moss) and rainwater collected in an undisturbed forest ecosystem. The (137)Cs activity and the isotopic ratio of (137)Cs/Cs in the samples were used to interpret the distribution and uptake of (137)Cs and the alkali metals in plants. As a whole, the (137)Cs in plants was assimilated together with K but was not dependent on Cs. Different adaptations of fern species collected in ecological niches cause them to have different (137)Cs/Cs ratios. Diplopterygium glaucum is distributed at the edges of the forest; it usually has shallow organic layers, and the root takes up more stable Cs from mineral layers, leading to lower (137)Cs/Cs ratios than that in the understory Plagiogyria formosana and Rhododendron formosanum species. The steady supply of stable Cs through the uptake by D. glaucum from deep soils may gradually dilute the (137)Cs concentration and thus explain the lower (137)Cs/Cs ratio in the fern samples. The (137)Cs is predicted to be proportional to the Cs content across plant species in the biological cycle once isotopic equilibrium is attained. PMID- 18417350 TI - Microwave-induced nucleophilic [18F]fluorination on aromatic rings: synthesis and effect of halogen on [18F]fluoride substitution of meta-halo (F, Cl, Br, I) benzonitrile derivatives. AB - The meta-halo-3-methylbenzonitrile derivatives (-F, -Cl, -Br, -I) were synthesized as model compounds to study reactivity towards aromatic nucleophilic substitution. A single-mode microwave system was incorporated into a commercial radiochemical synthetic module for (18)F labeling. Labeling yields of 64% for fluoro-, 13% for bromo- and 9% for chloro-precursors were achieved in DMSO in <3 min. The observed order of reactivity of the leaving groups toward aromatic nucleophilic substitution was F>>Br>Cl>>>I. PMID- 18417351 TI - In vivo gene delivery to proliferating cells in the striatum generated in response to a 6-hydroxydopamine lesion of the nigro-striatal dopamine pathway. AB - The degeneration of neurons in the mammalian brain is commonly associated with the division of cells located in the damaged area. The aim of the present study has been to characterise the phenotype of newly born cells in the striatum of adult rats following 6-hydroxydopamine lesion of the nigro-striatal pathway. Newborn cells were identified through labelling with either bromodeoxyuridine or retrovirus encoding green fluorescence protein. We report here that the overwhelming majority of these cells have glial characteristics. In order to promote the generation of new neurons we retrovirally introduced either the noggin or neurogenin2 genes into newborn cells following the 6-hydroxydopamine lesion. Transduction with neurogenin2 resulted in the production of cells resembling neuroblasts, however these cells did not appear to survive. Noggin transduction did not result in the generation of new neurons, but interestingly, greatly increased the number of oligodendrocytes generated from newborn cells. PMID- 18417353 TI - Development and optimization of an in vitro chloroplastic protein import assay using recombinant proteins. AB - The in vitro protein import experiment is one of the most important techniques for determining protein localization. For chloroplastic proteins, proteins of interest are incubated with isolated chloroplasts in the presence of energy sources. Radio-labeled proteins synthesized either in vitro or in vivo have been widely used as substrate proteins. Here we report our development of the protein import assay system in which non-radio-labeled proteins, overexpressed in Escherichia coli, were applied. In this system, substrate proteins were designed to carry epitope-tags, thus allowing analysis of imported proteins by SDS-PAGE, followed by immunoblotting to detect these tags. Furthermore, the imported proteins were found to be incorporated into their native form. These observations indicated that recombinant proteins were imported into chloroplasts and folded correctly. Therefore, this assay system could represent another valuable tool for determining protein localization. PMID- 18417352 TI - Expression of expanded polyglutamine targets profilin for degradation and alters actin dynamics. AB - Huntington's disease is caused by polyglutamine expansion in the huntingtin protein. Huntingtin directly interacts with profilin, a major actin monomer sequestering protein and a key integrator of signals leading to actin polymerization. We observed a progressive loss of profilin in the cerebral cortex of Huntington's disease patients, and in cell culture and Drosophila models of polyglutamine disease. This loss of profilin is likely due to increased degradation through the ubiquitin proteasome system. Profilin loss reduces the F/G actin ratio, indicating a shift in actin polymerization. Overexpression of profilin abolishes mutant huntingtin toxicity in cells and partially ameliorates the morphological and functional eye phenotype and extends lifespan in a transgenic polyglutamine Drosophila model. These results indicate a link between huntingtin and profilin and implicate profilin in Huntington's disease pathogenesis. PMID- 18417354 TI - Sun beds and cod liver oil as vitamin D sources. AB - The objective of this study was to (1) to determine the contribution of moderate sun bed exposure to serum 25(OH)D(3) levels; (2) to estimate the decay time of a high 25(OH)D(3) level obtained by sun bed exposure; and (3) to evaluate if the recommended ingestion of vitamin D is sufficient to maintain the 25(OH)D(3) concentration obtained by sun bed exposure. Ten volunteers (20-35 y.o.), skin type I and II, living in Olso, Norway were whole body exposed twice per week to the radiation of a commercial and approved sun bed (Life Sun S 100 W, Wolff System), starting with 0.5 MED (minimal erythema dose) and escalating to up to 1 MED per exposure for 4 weeks. After that, half of the volunteers were given a daily supplement of 200 IU vitamin D in the form of cod liver oil capsules, while the other half of the persons received no supplements. Erythema did not occur at any time and a slight pigmentation was seen in most of the volunteers after the sun bed exposures. Serum level of 25(OH)D(3) increased by about 40% on the average. The initial serum 25(OH)D(3) level was different among the volunteers (40-100 nmol/L). Within eight weeks after the last exposure the 25(OH)D(3) level decreased to the initial value in all volunteers irrespective of vitamin D supplementation or not. PMID- 18417355 TI - Risk stratification in neonates and infants submitted to cardiac surgery with cardiopulmonary bypass: a multimarker approach combining inflammatory mediators, N-terminal pro-B-type natriuretic peptide and troponin I. AB - Low cardiac output syndrome (LCOS) is a common problem following cardiac surgery with cardiopulmonary bypass (CPB) in neonates and infants, and its early recognition remains a challenging task. We aimed to test whether a multimarker approach combining inflammatory and cardiac markers provides complementary information for prediction of LCOS and death in children submitted to cardiac surgery with CPB. Forty-six children younger than 18 months with congenital heart defects were prospectively enrolled. No intervention was made. Blood samples were collected pre-operatively, during CPB and post-operatively (PO) for measurement of interleukin (IL)-6, IL-8, IL-10, tumor necrosis factor (TNF)-alpha, cardiac troponin I (cTnI) and N-terminal pro-B-type natriuretic peptide (NT-proBNP). Clinical data and outcome variables were recorded. Logistic regression was used to identify predictors of LCOS and death. Multivariate logistic regression identified pre-operative NT-proBNP and IL-8 4h PO as independent predictors of LCOS, while cTnI 4h PO and CPB length were independent predictors of death. The use of inflammatory and cardiac markers in combination improved sensitivity, negative predictive value and accuracy of the models. In conclusion, the combined assessment of inflammatory and cardiac biochemical markers can be useful for identifying young children at increased risk for LCOS and death after heart surgery with CPB. PMID- 18417356 TI - Disruption of the gamma c cytokine network in T cells during HIV infection. AB - The common gamma chain (gammac)-sharing cytokines (IL's-2, 4, 7, 9, 15, and 21) play a vital role in the survival, proliferation, differentiation and function of T lymphocytes. As such, disruption of their signaling pathways would be expected to have severe consequences on the integrity of the immune system. Indeed, it appears that the signaling network of these cytokines is both disrupted and exploited by HIV at various stages of infection. IL-2 secretion and signaling downstream of its receptor are impaired in T cells from chronically-infected HIV+ patients. Elevated plasma IL-7 levels and decreased IL-7Ralpha expression in patient T cells results in significantly decreased responsiveness to this critical cytokine. Interestingly, IL-2 and IL-15 are also able to render CD4+ T cells permissive to HIV infection through their influence on the activity of the APOBEC3G deaminase enzyme. Herein, we describe the current state of knowledge on how the gammac cytokine network is affected during HIV infection, with a focus on how this impairs CD4+ and CD8+ T cell function while also benefiting the virus itself. We also address the use of cytokines as adjuncts to highly active antiretroviral therapy to bolster immune reconstitution in infected patients. PMID- 18417357 TI - Characterization of phosphate-containing metabolites by calcium adduction and electron capture dissociation. AB - Several phosphate-containing metabolites, including nicotinamide adenine dinucleotide (NAD), nicotinamide adenine dinucleotide phosphate (NADP), adenosine 5'-diphosphate ribose (ADP-r), adenosine 5'-triphosphate (ATP), and guanosine 5' triphosphate (GTP), have been characterized with electron capture dissociation (ECD) and sustained off-resonance irradiation collision-activated dissociation (SORI-CAD) tandem mass spectrometry (MS/MS) in positive-ion mode. Calcium complexation was used to successfully produce abundant doubly charged cationic precursor ions with or without hydration. This approach enabled application of ECD to acidic metabolites for the first time. Fragmentation pathways observed in ECD and SORI-CAD of calcium-adducted phosphate-containing metabolites were complementary. Unique fragmentation was observed in ECD compared to SORI-CAD MS/MS, including ribose cross-ring cleavage for NAD and NADP, and generation of hydrated product ions, including cross-ring fragments, for hydrated ATP and GTP. A combination of ECD and CAD appears promising for maximizing structural information about metabolites. PMID- 18417359 TI - Strong anion exchange for studying protein-DNA interactions by H/D exchange mass spectrometry. AB - The use of mass spectrometry to study protein-ligand interactions is expanding into more complex systems including protein-DNA interactions. The excess amount of a model DNA or, more typically, an oligodeoxynucleotide (ODN), needed to study such interactions in an amide hydrogen-deuterium (H/D) exchange experiment, for example, causes serious signal suppression in the protein analysis. We describe here a modification of the traditional H/D exchange protocol whereby we utilize a strong anion exchange column to rapidly remove the ODN from solution before MS analysis. We showed the successful incorporation of such a column in a study of two protein-ODN systems: (1) the DNA-binding domain of human telomeric repeat binding factor 2 with a telomeric oligodeoxynucleotide and (2) thrombin with the thrombin-binding aptamer. The approach gave no appreciable difference in back exchange compared to a method in which no strong anion exchange (SAX) is used. PMID- 18417358 TI - On the benefits of acquiring peptide fragment ions at high measured mass accuracy. AB - The advantages and disadvantages of acquiring tandem mass spectra by collision induced dissociation (CID) of peptides in linear ion trap Fourier-transform hybrid instruments are described. These instruments offer the possibility to transfer fragment ions from the linear ion trap to the FT-based analyzer for analysis with both high resolution and high mass accuracy. In addition, performing CID during the transfer of ions from the linear ion trap (LTQ) to the FT analyzer is also possible in instruments containing an additional collision cell (i.e., the "C-trap" in the LTQ-Orbitrap), resulting in tandem mass spectra over the full m/z range and not limited by the ejection q value of the LTQ. Our results show that these scan modes have lower duty cycles than tandem mass spectra acquired in the LTQ with nominal mass resolution, and typically result in fewer peptide identifications during data-dependent analysis of complex samples. However, the higher measured mass accuracy and resolution provides more specificity and hence provides a lower false positive ratio for the same number of true positives during database search of peptide tandem mass spectra. In addition, the search for modified and unexpected peptides is greatly facilitated with this data acquisition mode. It is therefore concluded that acquisition of tandem mass spectral data with high measured mass accuracy and resolution is a competitive alternative to "classical" data acquisition strategies, especially in situations of complex searches from large databases, searches for modified peptides, or for peptides resulting from unspecific cleavages. PMID- 18417360 TI - Stable synaptic retention of serine-880-phosphorylated GluR2 in hippocampal neurons. AB - Phosphorylation of S880 within the GluR2 C-terminus has been reported to promote endocytosis of AMPA receptors (AMPARs) by preventing GluR2 interaction with the putative synaptic anchoring proteins GRIP and ABP. It is not yet established however, whether S880 phosphorylation induces removal of AMPARs from synaptic sites, and the trafficking of phosphorylated GluR2 subunits with surface and endocytosed GluR2 has not been directly compared within the same intact neurons. Here we show that phosphorylation of GluR2 subunits by PKC activated with phorbol esters is compartmentally restricted to receptors located at the cell surface. Endogenous AMPARs containing S880-phosphorylated GluR2 remained highly synaptic and colocalized with postsynaptic markers to the same extent as AMPARs which did not contain S880-phosphorylated GluR2. Moreover, following S880 phosphorylation, exogenous GluR2 homomers were found specifically at the cell surface and did not co-traffic with the internalized endosomal GluR2 population. We also show that GluR2 is endogenously phosphorylated by a constitutively active kinase pharmacologically related to PKC, and this phosphorylation is opposed by the protein phosphatase PP1. Our results demonstrate a population of hippocampal AMPARs which do not require interaction with GRIP/ABP for synaptic anchorage. PMID- 18417361 TI - The MUPP1-SynGAPalpha protein complex does not mediate activity-induced LTP. AB - At excitatory synapses of hippocampal neurons, the multi-PDZ domain scaffolding protein, MUPP1, assembles the NR2B subunit of the NMDA receptor (NMDAR), Ca2+ calmodulin kinase (CamKII), and the alpha1 isoform of the postsynaptic density GTPase activating protein, SynGAP (SynGAPalpha). In order to evaluate the role of this complex in excitatory synaptic neurotransmission we specifically disrupted MUPP1-SynGAPalpha interactions in CA1 neurons of acute hippocampal slices using intracellular perfusion with peptides derived from SynGAPalpha-MUPP1 binding domains. Disruption of the interaction between MUPP1 and SynGAPalpha with two complementary peptides derived from SynGAP and MUPP1 mutual binding sites resulted in enhancement of excitatory postsynaptic currents (EPSCs). This potentiation did not occlude pairing-induced long-term potentiation (LTP); indeed the amplitude of postsynaptic responses of activity-potentiated synapses was further increased. Pre-potentiation of excitatory synapses with theta burst stimulations did not modify the MUPP1-SynGAPalpha-dependent enhancement of EPSCs. Our data suggest that MUPP1-SynGAPalpha complex dissociation triggers a mechanism for AMPAR enhancement that is distinct from activity-induced LTP. PMID- 18417362 TI - Affective intentionality and self-consciousness. AB - We elaborate and defend the claim that human affective states ("feelings") are, among other things, self-disclosing. We will show why affective intentionality has to be considered in order to understand human self-consciousness. One specific class of affective states, so-called existential feelings, although often neglected in philosophical treatments of emotions, will prove central. These feelings importantly pre-structure affective and other intentional relations to the world. Our main thesis is that existential feelings are an important manifestation of self-consciousness and figure prominently in human self-understanding. We offer an ordering of four levels of existential feelings and also give considerations in favour of the essential bodily nature of these feelings. PMID- 18417363 TI - Elucidating the neural correlates of egoistic and moralistic self-enhancement. AB - Self-enhancement is the biasing of one's view of oneself in a positive direction. The brain correlates of self-enhancement remain unclear though it has been reported that the medial prefrontal cortex (MPFC) may be important for producing self-enhancing responses. Previous studies have not examined whether the neural correlates of self-enhancement depend on the particular domain in which individuals are enhancing themselves. Both moralistic and egoistic words were presented to participants while transcranial magnetic stimulation (TMS) was applied to the MPFC, precuneus or in a sham orientation. Participants were asked to make decisions as to the words describing themselves, some of which were positive and some of which were negative. It was found the MPFC TMS significantly disrupted egoistic self-enhancement when TMS was delivered to the MPFC. Judgments involving moralistic words were not influenced by TMS. These data provide further evidence that MPFC is involved in self-enhancement, and that the role of MPFC may be selective in this regard. PMID- 18417364 TI - Nicotine-induced nystagmus correlates with midpontine activation. AB - The pathomechanism of nicotine-induced nystagmus (NIN) is unknown. The aim of this study was to delineate brain structures that are involved in NIN generation. Eight healthy volunteers inhaled nicotine in darkness during a functional magnetic resonance imaging (fMRI) experiment; eye movements were registered using video-oculography. NIN correlated with blood oxygen level-dependent (BOLD) activity levels in a midpontine site in the posterior basis pontis. NIN-induced midpontine activation may correspond to activation of the dorsomedial pontine nuclei and the nucleus reticularis tegmenti pontis, structures known to participate in the generation of multidirectional saccades and smooth pursuit eye movements. PMID- 18417365 TI - Differential superior prefrontal activity on initial versus subsequent shifts in naive subjects. AB - Flexible adaptation to changing environments requires shifting of a cognitive set, one basic function of the prefrontal cortex. Set shifting, as instantiated in the Wisconsin Card Sorting Task (WCST) administered in a neuropsychological testing room, is typically achieved when subjects have no prior experiences of updating one WCST behavior to another. By contrast, earlier neuroimaging studies typically involved examination of repeated transitions between particular behaviors, to which situation subjects are far from naive. Naive subjects with no prior knowledge of the WCST were recruited in the present functional magnetic resonance imaging study to test set shifting under unknown situations that they experienced for the first time. Prominent activation was revealed in the left superior prefrontal cortex selectively on the initial shifts. On the other hand, the inferior prefrontal cortex was significantly activated on both the initial and subsequent shifts. The superior prefrontal activation distinguishable from the conventional inferior prefrontal activation suggests a selective role of this region in performance of the WCST in naive subjects. PMID- 18417366 TI - Folic acid and methionine in the prevention of teratogen-induced congenital defects in mice. AB - INTRODUCTION: Periconceptional supplementation with multivitamins containing folic acid reduces the risk of congenital malformations. We have previously investigated the effect on the murine development of a multiple retinoic acid competitive antagonist, Bristol-Myers-Squibb 189453, showing that treated fetuses were affected with heart defects, thymus aplasia or hypoplasia, and severe anomalies of the central nervous system. Hereby, we analyzed the effects of nutritive therapy involving folic acid and methionine on teratogen-induced congenital defects in mice. MATERIALS AND METHODS: A total of 132 outbred CD1 litters were studied. Pregnant mice were divided into four experimental groups, and an oral supplementation of H(2)O or folic acid, or methionine, or folic acid+methionine was administered from 0.5 days postcoitum until the end of pregnancy. At 7.5 days postcoitum, mice from all these groups were administered Bristol-Myers-Squibb 189453 to induce the teratogenic effect. At the end of pregnancy, fetuses were dissected and tissues were analyzed by histology and flow cytometric assays. RESULTS: Folic acid reduces congenital heart diseases from 81.3% to 64.8%, neural tube defects from 20.3% to 3.7%, and thymus abnormalities from 98.4% to 27.8%, restoring a normal number of differentiated thymus cells. Methionine is less effective in contrasting congenital heart diseases and neural tube defects, and induces thymus cell proliferation but not differentiation. Folic acid+methionine weakly reduce congenital heart diseases and neural tube defects, but consistently reduce the incidence of fetuses affected with thymus pathologies from 98.4% to 67.7%. CONCLUSIONS: Our results suggest that folic acid and methionine periconceptional supplementations may influence the incidence of congenital defects and may probably induce negative selection of embryos presenting developmental anomalies. PMID- 18417367 TI - Presence of fatty-acid-binding protein 4 expression in human epicardial adipose tissue in metabolic syndrome. AB - BACKGROUND: Metabolic syndrome is a cluster of different clinical manifestations that are risk factors for atherothrombotic cardiovascular disorders. Fatty-acid binding protein 4 (FABP4/aP2), which is highly expressed in adipocytes, specifically exerts intracellular lipid trafficking. A high level of fatty-acid binding protein 4 expression present in obese subjects has also been found in mice and humans, especially in macrophages at atherosclerotic lesions. An in vivo study demonstrated that the inhibitor of aP2 would be a new therapeutic agent for treating metabolic diseases in mice. We have investigated the mRNA expression of fatty-acid-binding protein 4 in human epicardial adipose and ascending aorta tissues of metabolic syndrome and nonmetabolic syndrome patients. METHODS: Paired epicardial adipose and ascending aorta tissue samples were obtained from 10 metabolic syndrome patients and 4 nonmetabolic syndrome patients during coronary bypass grafting and aortic valve replacement therapy, respectively. Fatty-acid binding protein 4 gene expression was determined by quantitative real-time polymerase chain reaction. RESULTS AND CONCLUSIONS: Fatty-acid-binding protein 4 expression of epicardial adipose tissue was significantly higher in metabolic syndrome patients than in nonmetabolic syndrome controls (P<.05). In metabolic syndrome patients, fatty-acid-binding protein 4 expression in epicardial adipose tissue was 66 times higher than fatty-acid-binding protein 4 expression in ascending aorta tissue. The expression level of fatty-acid-binding protein 4 in epicardial adipose tissue was found to be significantly correlated with waist circumference in all subjects (r=.535, P<.05). Our data showed for the first time that human epicardial adipose and ascending aorta tissues express fatty-acid binding protein 4 and that its level of expression in epicardial adipose tissues of metabolic syndrome patients is elevated. Increased fatty-acid-binding protein 4 gene expression in epicardial adipose tissues of metabolic syndrome patients led us think that fatty-acid-binding protein 4 might be an important factor in metabolic syndrome. PMID- 18417368 TI - Intravascular epithelioid hemangioendothelioma of the inferior vena cava: case report of an unusual and unpredictable vascular tumor. AB - Vascular neoplasms are uncommon and pose a diagnostic and treatment challenge to the pathologist and surgeon, respectively. Epithelioid hemangioendothelioma is a rare neoplasm of vascular origin with an unknown etiology. Its biologic behavior lies somewhere between that of a benign hemangioma and that of a malignant angiosarcoma; however, it is unpredictable at best. Intravascular epithelioid hemangioendotheliomas have been described more often in veins than arteries, and there are only about 30 reports in the English literature. We report here the case of an epithelioid hemangioendothelioma of the inferior vena cava, which presented with abdominal pain, ascites and pedal edema. PMID- 18417369 TI - Cardiac rhabdomyoma. AB - Primary tumors of the heart are exceedingly rare with a prevalence rate of around 0.01% in autopsy studies. A majority of primary cardiac tumors are benign. Rhabdomyomas are the most common of benign pediatric cardiac tumors. A newborn female infant presented with perioral cyanosis and respiratory distress. She deteriorated progressively. The infant expired, and at autopsy, dissection of the heart revealed a large tumor mass in the interventricular septum. Histologically, the mass showed the classic "spider cells," and a diagnosis of cardiac rhabdomyoma of childhood was established. PMID- 18417370 TI - The Cryptocercus punctulatus species complex (Dictyoptera: Cryptocercidae) in the eastern United States: comparison of cuticular hydrocarbons, chromosome number, and DNA sequences. AB - The goal of the current study was to determine if cuticular hydrocarbons could be used to empirically delimit taxa within the Cryptocercus punctulatus species complex in the eastern United States. Cockroaches were collected from rotting logs in 22 locations across four states. Hydrocarbon phenotypes and two mitochondrial (16S and COII) genes and one nuclear (ITS2) gene were independently analyzed to determine their relationship with chromosome number. Five distinct hydrocarbon phenotypes were found, but these were only partly congruent with chromosome number and thus with purported species descriptions. Molecular and cuticular hydrocarbon data each indicate that Cryptocercus with a male karyotype of 2n=43 belong to at least two distinct, distantly related lineages. One 2n=43 lineage is sister group to the 2n=37 and 2n=39 clade, and has a unique hydrocarbon profile. The other 2n=43 lineage is sister group to the 2n=45 samples, and its cuticular hydrocarbons group with four samples of the 2n=45 lineage. The cuticular hydrocarbons of two additional 2n=45 samples diverge from this assemblage. Results indicate cuticular hydrocarbons and chromosome number have some degree of evolutionary independence; neither is completely reliable in delineating historical lineages. Our data provide support for the parallel model of chromosome evolution in the species complex. PMID- 18417371 TI - How many ECG leads are required for in vivo studies in safety pharmacology? AB - This article explains the principles of electrocardiography, and explains how it is used by Safety Pharmacology, with a focus on the requirement for multiple leads in Safety Pharmacology assessment. Electrocardiography as used in different disciplines (e.g., medicine, anesthesiology, physiology, and pharmacology/toxicology/safety pharmacology) has different requirements for the number of electrodes applied. Electrodes may be placed at an infinite number of points on the body, and voltages (electrocardiograms) may be registered between/among them. However in safety pharmacology there is little evidence that more than 1--or at most 3--lead(s) is (are) required to provide all of the information that might be present using an infinite number. This is based upon (1) the biophysics of the heart as a generator of electrical potential/voltage, (2) the fact that most properties of electrophysiology affected adversely by drugs are expressed as changes in durations, and (3) experience. A single, unipolar lead (V(3)) recorded from the left sternal border at the 5th intercostal space possesses minimal artifact and large, stable deflections. This lead allows for accurate measurement of heart rate and rhythm, durations of component deflections (e.g., PQ, QRS, QT), and J-point deviation. A greater number of leads seldom or never yield additional information that detects liabilities. Commonly voltages recorded between the right thoracic and left pelvic limbs (lead II) provides information similar to lead V(3), and lead II is easier to apply, and produces voltages with less artifact and similar to those in lead V(3). A lead measuring the voltage between the left and right thoracic limbs (lead I) along with lead II allows for estimating orientation of vectors in the frontal plane, but knowledge of these vectors seldom or never indicates liability of a test article. PMID- 18417372 TI - Recovery from stroke in patients with diabetes mellitus. AB - BACKGROUND AND AIM: Diabetes mellitus (DM) is recognized as an important risk factor for stroke and might theoretically influence post-stroke level of disability, increasing the extension of the cerebral injured area. However, results of the few researches aimed at studying this influence are contradictory; moreover, the effect of DM on motor recovery has not been extensively studied. The aim of this study was to investigate the effect of DM on both functional and motor recovery. METHODS: A total of 395 acute patients with first stroke were selected in a rehabilitation department and divided into two groups on the basis of the presence or absence of DM (DM+ and DM-, respectively). Outcome measures were the Barthel Index, the Fugl-Meyer Assessment Scale, and the mobility part of the motor assessment chart according to Lindmark and Hamrin. Participants were assessed at admission to department (T1, 13.9+/-7.9 days from stroke onset), at discharge (T2, 40.1+/-13.4), and at follow-up (T3, 84.2+/-14.3). A 2 x 3 analysis of variance with repeated measures was performed to verify the effect of group and of phase of assessment on motor and functional measures and their interaction. RESULTS: DM+ and DM- groups included 93 and 302 patients, respectively. Both groups showed a significant and progressive improvement in all outcome measures (P<.001), but no interaction was found between group and phase of assessment, which means that motor and functional recovery was similar in the two groups. CONCLUSION: Results suggest that diabetes has no influence on motor and functional outcome within the acute and post-acute phase after stroke. Further research should investigate motor recovery in a longer-term period and with larger samples. PMID- 18417373 TI - Comparative evaluation of three semi-quantitative radiographic grading techniques for knee osteoarthritis in terms of validity and reproducibility in 1759 X-rays: report of the OARSI-OMERACT task force. AB - OBJECTIVE: The objective of this work was to compare the measurement properties of three categorical X-ray scoring methods of knee osteoarthritis (OA), both on semiflexed and extended views. METHODS: In data obtained from trials and cohorts, X-rays were graded using Kellgren and Lawrence (KL), the OA Research Society International (OARSI) joint space narrowing score, and measurement of joint space width (JSW). JSW was analyzed as a categorical variable. Construct validity was assessed through logistic regression between X-ray stages and Western Ontario and McMaster Universities OA Index. Inter-observer reliability was assessed in 50 subjects for extended views by weighted kappa. Intra-observer reliability and sensitivity to change were assessed separately for extended and semiflexed views in 50 patients who had both views performed, over a 30-month interval, by weighted kappa and standardized response mean (SRM). RESULTS: Extended views were available from three trials and two cohorts (1759 X-rays), including one trial in which both extended and semiflexed views (antero-posterior) were obtained. Correlation with clinical parameters was low for the three scoring methods, except for the single community-based cohort. Inter-rater reliability was higher for categorical JSW in extended views (kappa, 0.86 vs 0.56 and 0.48 for KL and OARSI, respectively). Intra-rater reliability was higher for categorical JSW, both in extended views (0.83 vs 0.61 and 0.71) and in semiflexed views (0.89 vs 0.50 and 0.67). Sensitivity to change was also higher for categorical JSW, particularly in semiflexed views (SRM, 0.49 vs 0.22 and 0.34). CONCLUSION: These results indicate categorical JSW, in particular on semiflexed views, may be the preferred method to evaluate structural severity in knee OA clinical trials. PMID- 18417375 TI - Systematic review of the nutritional supplements dimethyl sulfoxide (DMSO) and methylsulfonylmethane (MSM) in the treatment of osteoarthritis. AB - OBJECTIVE: Conventional treatment of osteoarthritis (OA) with non-steroidal anti inflammatory drugs is associated with serious gastrointestinal side effects and in view of the recent withdrawal of some cyclo-oxygenase-2 inhibitors, identifying safer alternative treatment options is needed. The objective of this systematic review is to evaluate the existing evidence from randomised controlled trials of two chemically related nutritional supplements, dimethyl sulfoxide (DMSO) and methylsulfonylmethane (MSM) in the treatment of OA to determine their efficacy and safety profile. METHODS: The electronic databases [Cochrane Library, Medline, Embase, Amed, Cinahl and NeLH (1950 to November 2007)] were searched. The search strategy combined terms: osteoarthritis, degenerative joint disorder, dimethyl sulfoxide, DMSO, methylsulfonylmethane, MSM, clinical trial; double blind, single blind, RCT, placebo, randomized, comparative study, evaluation study, control. Inclusion and exclusion criteria were applied. Data were extracted and quality was assessed using the JADAD scale. RESULTS: Six studies were included [evaluating a total of 681 patients with OA of the knee for DMSO (N=297 on active treatment); 168 patients for MSM (N=52 on active treatment)]. Two of the four DMSO trials, and both MSM trials reported significant improvement in pain outcomes in the treatment group compared to comparator treatments, however, methodological issues and concerns over optimal dosage and treatment period, were highlighted. CONCLUSION: No definitive conclusion can currently be drawn for either supplement. The findings from all the DMSO studies need to be viewed with caution because of poor methodology including; possible unblinding, and questionable treatment duration and dose. The data from the more rigorous MSM trials provide positive but not definitive evidence that MSM is superior to placebo in the treatment of mild to moderate OA of the knee. Further studies are now required to identify both the optimum dosage and longer-term safety of MSM and DMSO, and definitive efficacy trials. PMID- 18417374 TI - Histone deacetylase inhibitors suppress interleukin-1beta-induced nitric oxide and prostaglandin E2 production in human chondrocytes. AB - OBJECTIVE: Overproduction of nitric oxide (NO) and prostaglandin E(2) (PGE(2)) plays an important role in the pathogenesis of osteoarthritis (OA). In the present study, we determined the effect of trichostatin A (TSA) and butyric acid (BA), two histone deacetylase (HDAC) inhibitors, on NO and PGE(2) synthesis, inducible NO synthase (iNOS) and cyclooxygenase (COX)-2 expression, and nuclear factor (NF)-kappaB DNA-binding activity, in interleukin-1beta (IL-1)-stimulated human OA chondrocytes, and on IL-1-induced proteoglycan degradation in cartilage explants. METHODS: Chondrocytes were stimulated with IL-1 in the absence or presence of increasing concentrations of TSA or BA. The production of NO and PGE(2) was evaluated using Griess reagent and an enzyme immunoassay, respectively. The expression of iNOS and COX-2 proteins and mRNAs was evaluated using Western blotting and real-time reverse transcriptase-polymerase chain reaction (RT-PCR), respectively. Proteoglycan degradation was measured with dimethymethylene blue assay. Electrophoretic mobility shift assay (EMSA) was utilized to analyze the DNA-binding activity of NF-kappaB. RESULTS: HDAC inhibition with TSA or BA resulted in a dose-dependent inhibition of IL-1-induced NO and PGE(2) production. IL-17- and tumor necrosis factor-alpha (TNF-alpha) induced NO and PGE(2) production was also inhibited by TSA and BA. This inhibition correlated with the suppression of iNOS and COX-2 protein and mRNA expression. TSA and BA also prevented IL-1-induced proteoglycan release from cartilage explants. Finally, we demonstrate that the DNA-binding activity of NF kappaB, was induced by IL-1, but was not affected by treatment with HDAC inhibitors. CONCLUSIONS: These data indicate that HDAC inhibitors suppressed IL-1 induced NO and PGE(2) synthesis, iNOS and COX-2 expression, as well as proteoglycan degradation. The suppressive effect of HDAC inhibitors is not due to impaired DNA-binding activity of NF-kappaB. These findings also suggest that HDAC inhibitors may be of potential therapeutic value in the treatment of OA. PMID- 18417376 TI - Painful tongue ulcerations in patients with bisphosphonate-associated osteonecrosis of the jaws. AB - OBJECTIVES: Bisphosphonate-associated osteonecrosis of the jaws (BONJ) is a recently recognized and well described complication in cancer patients. While pain and associated morbidity have been reported, this has largely been attributed to localized infection of the nonvital bone and surrounding soft tissue, or in rare cases, pathologic fracture. STUDY DESIGN: Cancer patients who were treated with intravenous bisphosphonate therapy and subsequently developed BONJ with associated traumatic tongue ulcerations were reported. RESULTS: We report a series of 5 patients who presented with unilateral painful tongue ulcerations secondary to the tongue rubbing against adjacent exposed mylohyoid bone on the lingual surface of the mandible. Smoothing, recontouring, or removal of the bone, and in some cases intralesional steroid therapy, resulted in complete resolution of symptoms. CONCLUSIONS: Oral health care specialists and physicians must be alert to this potential complication of BONJ because it is easily managed with conservative measures and results in rapid and significant improvement in symptoms and overall quality of life. PMID- 18417377 TI - Ameloblastoma associated with the nevoid basal cell carcinoma (Gorlin) syndrome. AB - Nevoid basal cell carcinoma syndrome (NBCCS) is an autosomal dominant disorder characterized by a wide range of clinical signs and symptoms. The major criteria for the diagnosis are multiple cutaneous basal cell carcinomas, multiple odontogenic keratocysts of the jaw, palmar and plantar pits, and skeletal abnormalities. Here, we report an unusual case of NBCCS in a 68-year-old woman with late onset of clinical signs and symptoms and with an associated ameloblastoma. Only 4 other cases of this unusual association have been reported. PMID- 18417378 TI - Odontogenic tumors: a study of 250 cases in an Indian teaching hospital. AB - OBJECTIVE: Studies on odontogenic tumors published from many parts of the world show a distinct geographic variation. But there is little information available in the English-language literature on the relative frequency of odontogenic tumors in India. This retrospective study was designed to determine the relative frequency of odontogenic tumors in an Indian teaching hospital and compare them with various reports from other parts of the world. STUDY DESIGN: The pathology records of the Department of Oral Pathology, Nair Hospital Dental College, from January 1971 to December 2006 were reviewed retrospectively, and based on World Health Organization classifications, 1992, we reclassified and analyzed the odontogenic tumors. The mandible and maxilla were divided into 7 anatomic regions, and the distribution of each odontogenic tumor among these regions was recorded and analyzed. RESULTS: A total of 250 cases of odontogenic tumors were reported in the 36-year period. Of these cases, 98.8% of the tumors were benign. The most frequent histologic type was ameloblastoma (61.5%), followed by adenomatoid odontogenic tumor (12.4%) and myxomas (6%). Odontomas, which are reported as the most frequent odontogenic tumor in American and European case reports, only accounted for 6%. In general, the odontogenic tumors showed a predilection for the mandible and the posterior regions of the jaws. Ameloblastomas occurred with a marked predilection for the mandible with a ratio of 18.1:1. But adenomatoid odontogenic tumor showed predilection for the maxilla, anterior regions of the jaws, and young females. CONCLUSIONS: Odontogenic tumors show a definite geographic variation. In India, ameloblastoma and adenomatoid odontogenic tumor are the most frequent odontogenic tumors, with distinct anatomic predilections. PMID- 18417379 TI - Heterotopic neuroglial tissue: two cases involving the tongue and the buccal region. AB - We report two rare cases of extranasal heterotopic neuroglial tissue and review the literature on the topic. The clinical, histological, and immunohistochemical features are presented. Both patients had lesions extranasally, even though the sinonasal region is the predominant site for these heterotopias. The first lesion was located in the buccal area in an 8-year-old boy and the second lesion in the tongue of a 2-year-old boy. They had relatively small lesions with few clinical symptoms. Complete excision was made and the follow-up was unremarkable. Heterotopic neuroglial tissue is considered to be a congenital condition. Complete excision of these lesions is recommended in spite of the lack of malignant potential, but because of the possibility of continuous growth. PMID- 18417380 TI - Dedifferentiated epithelial-myoepithelial carcinoma of the parotid gland: a rare case report of immunohistochemical analysis and review of the literature. AB - Dedifferentiation of salivary gland neoplasms is a rare event, unlike bone and soft part sarcomas, which was first described by Stanley et al. in 1988. An additional case of dedifferentiated epithelial-myoepithelial carcinoma (EMC) is reported here. The patient was a 70-year-old Japanese man who requested examination of the rapid growth of a mass in the right parotid region, which he had first noticed 25 years previously. Clinical examination showed an ill circumscribed, 6.8 x 4.7 x 7.0-cm lesion. Histologically, most parts of the lesion were high-grade carcinoma (HGC) with sheetlike and nestlike growth of markedly atypical cells and comedonecrosis, whereas the minor part consisted of typical EMC. The outer clear cells of EMC were positive for alpha-smooth muscle actin (ASMA), p63, cytokeratin (CK) 14, and vimentin, and the inner ductal cells of EMC were positive for CKs and epithelial membrane antigen. HGC was negative for ASMA, CK14, and vimentin, but diffusely positive for p53 protein and cyclin D1. The Ki-67 labeling index of EMC was 11.5%, whereas that of HGC was 67.1%. These findings and a review of literature indicate that HGC arose from preexisting EMC, and this phenomenon is the dedifferentiation of EMC. Dedifferentiated EMC is extremely rare. PMID- 18417381 TI - Condylar positioning devices for orthognathic surgery: a literature review. AB - In the past few years, many devices have been proposed for preserving the preoperative position of the mandibular condyle during bilateral sagittal split osteotomy. Accurate mandibular condyle repositioning is considered important to obtain a stable skeletal and occlusal result, and to prevent the onset of temporomandibular disorders (TMD). Condylar positioning devices (CPDs) have led to longer operating times, the need to keep intermaxillary fixation as stable as possible during their application, and the need for precision in the construction of the splint or intraoperative wax bite. This study reviews the literature concerning the use of CPDs in orthognathic surgery since 1990 and their application to prevent skeletal instability and contain TMD since 1995. From the studies reviewed, we can conclude that there is no scientific evidence to support the routine use of CPDs in orthognathic surgery. PMID- 18417382 TI - Association between salivary flow rates, oral symptoms, and oral mucosal status. AB - OBJECTIVE: To investigate the association of salivary flow rates with oral symptoms and oral mucosal status. STUDY DESIGN: The study population included 462 Israeli subjects attending a xerostomia clinic. After patient history and oral mucosal examination, major gland sialometry, and complementary tests, patients were divided into 6 groups: drug-induced salivary gland hypofunction (SGH), Sjogren syndrome (SS), radiation-induced SGH, idiopathic SGH, xerostomia without SGH, and control. RESULTS: Oral mucosal alterations were more prevalent in all SGH groups than in the control group. Oral symptoms (except speech impairment) were more frequent in all SGH groups. The postradiation group showed the highest frequency of oral mucosal alterations and of swallowing and mastication complaints. Individuals complaining of xerostomia (compared with those who did not) displayed lower major salivary gland flow rates and a higher frequency of oral mucosal alterations CONCLUSIONS: Presence of oral mucosal alterations may help but are not enough to identify patients for further evaluation of SGH. Difficulties in mastication and swallowing are most specifically related to advanced SGH. PMID- 18417383 TI - Do oral lesions associated with HIV affect quality of life? AB - OBJECTIVE: To assess the impact of oral lesions on quality of life (QOL) in HIV positive (HIV+) individuals using the Oral Health Impact Profile (OHIP). STUDY DESIGN: This was a cross-sectional analytic study that compared 2 groups of HIV+ patients. Group 1 (n = 71) consisted of patients who presented with oral manifestations of HIV infection. Patients in group 2 (n = 79) were also HIV positive but had no oral lesions. The QOL measures were assessed by either simple frequency counts or the use of item weights. RESULTS: Oral candidiasis was the most common oral lesion diagnosed. Patients in group 1 reported significantly greater impacts (P < .05) for all 7 subscales in OHIP. CONCLUSIONS: Oral lesions associated with HIV infection negatively affect the oral health-related QOL in infected patients. PMID- 18417384 TI - In vitro evaluation of the sealing ability of MTA used for the repair of furcation perforations with and without the use of an internal matrix. AB - OBJECTIVE: To evaluate the effect of matrix on mineral trioxide aggregate (MTA) when used to repair furcal perforations. STUDY DESIGN: Forty-five human molars with furcal perforations were randomly divided into 3 groups and repaired with MTA either alone or over a barrier (calcium sulfate or Collaplug). Eight teeth without perforations provided negative control samples. After assessing repair quality, 1 mol/L glucose solution was used as the tracer to detect the leakage. The concentration of leaked glucose was measured after 1, 2, 4, 7, 15, and 20 days using a glucose oxidase method. RESULTS: No significant difference in either leakage or overfilling was found between the groups with Collaplug and MTA alone (P > .05). Although no overfilling was detected in the group with calcium sulfate, that group had the most leakage (P < .05). CONCLUSIONS: Neither of the 2 internal matrices improved the sealing ability and reduced the incidence of overfilling simultaneously. PMID- 18417385 TI - Oral myofibromas: report of two cases and review of clinical and histopathologic differential diagnosis. AB - Myofibroma is a benign mesenchymal neoplasm composed of myofibroblasts which has been described with different synonyms since the first report in 1951. It may show clinical and histologic features that may be misinterpreted as a malignancy. We describe 2 cases of oral myofibromas affecting infants; the first one showed a rapid growth with teeth displacement and ulceration; the second one presented a relatively slow growth with an indolent course. Differential diagnosis included benign and malignant mesenchymal neoplasms, salivary gland tumors, and reactive processes. Microscopic analysis of both lesions revealed a spindle cell tumor with immunoreactivity for vimentin, muscle-specific actin, and specific smooth muscle isoform alpha-actin, rendering the diagnoses of myofibroma. The patients were treated with surgical excision, and both are in follow-up without any signs of recurrence. Myofibroma presents a wide range of differential diagnosis, including benign and malignant neoplasms. Therefore, accurate diagnosis may avoid an unnecessary aggressive therapy. PMID- 18417386 TI - Radiological evaluation of changes in the gonial angle after teeth extraction and wearing of dentures: a 3-year longitudinal study. AB - OBJECTIVE: The aim of this study was to evaluate the gonial angle measurements of 20 adult patients made from panoramic radiographs and to assess differences from dentate state to edentate state-with dentures. STUDY DESIGN: A total of 100 panoramic radiographs of 20 patients were evaluated, and gonial angle sizes were measured. Panoramic radiographs were made while the patients were dentulous and in the first month, sixth month, first year, and third year after tooth extraction and wearing dentures after a 2-3-month healing period. All radiographs used in this study were made with the same panoramic machine by the same person. Gonial angles were measured by tracing a line on panoramic radiographs tangential to the most inferior points at the gonial angle and the lower border of the mandibular body and another line tangential to the posterior borders of the ramus and the condyle. RESULTS: Difference in size of the gonial angle was found for both sides between continuous measurements. The gonial angle values tend to increase in both sides after tooth extractions. After the third measurement for both sides, the values start to decrease. CONCLUSION: After wearing denture for 3 years, a statistically significant decrease was found in the gonial angle values according to the edentulous state (P < .05). PMID- 18417387 TI - Osteolipoma of the oral and pharyngeal region: report of a case and review of the literature. AB - Osteolipoma is an uncommon phenomenon that rarely occurs in the oral and pharyngeal region. A 68-year-old man presented with a 4-year history of a relatively small painless mass in the right mandibular alveolar mucosa adjacent to the premolar teeth. The lesion was easily excised and histologically confirmed to be an osteolipoma. Ten cases of oral and pharyngeal osteolipoma have appeared in the English literature thus far. In addition to reporting a new case of osteolipoma arising in the oral cavity in this article, the literature is reviewed accordingly. PMID- 18417388 TI - Mechanical properties of a new and improved nickel-titanium alloy for endodontic use: an evaluation of file flexibility. AB - OBJECTIVE: A new manufacturing method to produce more flexible and resistant NiTi endodontic instruments has been recently developed. The purpose of the study was to determine whether this new manufacturing method produces NiTi instruments (TF) of superior flexibility. STUDY DESIGN: Ten .06 size 25 prototype (TF) instruments (Sybron Endo) and ten 06.25 ProFile instruments (Dentsply-Maillefer) were evaluated for stiffness on bending. Test procedures strictly followed ANSI/ADA Specification No.28. Bending moment was measured when the instrument attained a 45 degrees bend. All data obtained were recorded and statistically analyzed using an ANOVA test. RESULTS: Statistical analysis of data showed that bending moments were significantly greater (P < .05) for ProFile instruments (mean value 59.06 g/cm), compared to the prototype instruments (mean value 35.60 g/cm), thereby confirming the increased flexibility of TF instruments. CONCLUSION: Results of the present study demonstrate the ability of the new manufacturing process to produce more flexible NiTi rotary instruments. PMID- 18417389 TI - Prevalence of commonly found pathoses associated with mandibular impacted third molars based on panoramic radiographs in Turkish population. AB - OBJECTIVE: The objective was to determine the association between commonly found pathologic conditions and angulations and impaction depths of lower third molar teeth. STUDY DESIGN: In this retrospective study 1,914 panoramic radiographs with 3,050 impacted lower third molars (ILTMs) were examined. RESULTS: The problems and the prevalence evaluated were caries on mandibular second molar (12.6%), caries on ILTM (5.3%), bone loss at the distal aspect of ILTM (9.7%), and periodontal tissue damage on adjacent tooth (8.9%). A total of 26.5% of all the cases were affected by any one of the 4 pathologic changes. It was determined that horizontal and mesioangular ILTMs had high risk of caries development on second or third molar; in contrast, vertical and distoangular ILTMs had low risk. Distoangular and vertical ILTMs had high risk of development of bone loss at the distal aspect. Mesioangular and horizontal ILTMs had a high risk for periodontal tissue damage on an adjacent tooth. When evaluating impaction depth of the ILTM, class A had a higher risk of pathology than class B or C. CONCLUSIONS: Horizontal and mesioangular impactions were found with more pathologic situations; especially in class A impaction depth. Angulation and impaction depth of the ILTM should be taken into consideration when making a decision whether to extract an ILTM or not. PMID- 18417390 TI - Oral malodor and removable complete dentures in the elderly. AB - OBJECTIVE: The objectives of this study were to assess oral malodor in patients with removable complete dentures and analyze its association with age, sex, subjective oral dryness, burning mouth, tongue coating, and overnight denture wear in elderly subjects. STUDY DESIGN: The study comprised 105 complete denture wearers (56 male, 49 female) between the ages of 50 and 78 from the province of Kirikkale in the Central Anatolia Region of Turkey. Oral malodor was assessed using a portable sulfide monitor. Participants were given a self-administered, structured questionnaire. Baseline and final volatile sulfur compound (VSC) levels and tongue-coating scores (TCS) were measured. Analysis included frequency, cross-tabulations, ANOVA, and independent sample and paired t tests. Odds ratios of VSC-related factors were also calculated. A P value less than .05 was considered statistically significant. RESULTS: There was no significant correlation found between age, gender, smoking, existence of a medical condition, or regular use of medication and VSC levels in mouth air (P > .05). However, educational level (P < .05), oral dryness (P < .05), burning mouth (P < .05), overnight denture wear (P < .01), and TCS (P < .001) were found to be significantly correlated with VSC levels. Significant differences were also found between baseline and final measurements of both VSC levels and TCS (P < .01). CONCLUSION: This study found the accumulation of bacterial plaque on the tongue, oral dryness, burning mouth, overnight denture wear, and lower educational levels to be significantly related to oral malodor. Following tongue care maintenance and overnight removal of dentures, patients' oral malodor levels decreased significantly. PMID- 18417391 TI - Getting published--improving the odds. PMID- 18417392 TI - Characteristics of 283 supernumerary teeth in southern Chinese children. AB - OBJECTIVES: This was a retrospective study to determine the characteristics and distribution of supernumerary teeth in southern Chinese children. METHODS: The study population consisted of 208 children aged from 2 to 16 years. RESULTS: Males were more frequently affected than females in the ratio 3.1:1. Of the 283 supernumerary teeth, 95.0% were located in the premaxilla, 71.5% were conical, 70.7% were unilateral, 29.3% were bilateral, 47.7% were inverted, and 16.9% were erupted. The mean age at the time of diagnosis and removal of the supernumerary teeth was 7.3 +/- 2.7 years (the minimum age was 2.1 years) and 8.1+/- 2.7 years (the minimum age was 4.1 years), respectively. Seventy percent of the children were in the mixed dentition and 81.3% of the supernumeraries were removed under general anesthesia. CONCLUSION: In accordance with previous studies, the majority of the supernumerary teeth were located in the premaxillary region, were conical shaped, and remained unerupted. PMID- 18417393 TI - Nasopharyngeal stenosis secondary to soft palate dysgenesis in a cat. AB - This report is the first description of soft palate dysgenesis inducing nasopharyngeal stenosis in a cat. The symptomatology included continuous stertorous respiration and changes in phonation. Open-mouth breathing, seromucous nasal discharge and sneezing were intermittently present. On physical examination the cat showed a respiratory pattern characteristic of upper airway disease, with stridor, increased inspiratory effort and severe bilateral obstruction of nasal passages to the airflow. Pharyngo-laryngoscopic evaluation showed an abnormal conformation of the soft palate that appeared attached to the caudal and lateral pharyngeal wall and stretched the whole length of the pharynx. A strip of soft tissue emanated from the caudal part of the hard palate to the right aspect of the base of the tongue. Caudal to this strip of tissue, the intrapharyngeal opening was stenosed, therefore preventing normal airflow. Surgical reconstruction was not recommended. The cat was clinically stable with mild respiratory symptoms 17 months after the diagnosis. PMID- 18417394 TI - On the measurement of 15N-{1H} nuclear Overhauser effects. AB - Accurate quantification of the 15N-{1H} steady-state NOE is central to current methods for the elucidation of protein backbone dynamics on the fast, sub nanosecond time scale. This experiment is highly susceptible to systematic errors arising from multiple sources. The nature of these errors and their effects on the determined NOE ratio is evaluated by a detailed analysis of the spin dynamics during the pair of experiments used to measure this ratio and possible improvements suggested. The experiment that includes 1H irradiation, is analyzed in the framework of Average Liouvillian Theory and a modified saturation scheme that generates a stable steady-state and eliminates the need to completely saturate 1H nuclei is presented. The largest source of error, however, in 1H dilute systems at ultra-high fields is found to be an overestimation of the steady-state NOE value as a consequence of the incomplete equilibration of the magnetization in the so-called "reference experiment". The use of very long relaxation delays is usually an effective, but time consuming, solution. Here, we introduce an alternative reference experiment, designed for larger, deuterated systems, that uses the fastest relaxing component of the longitudinal magnetization as a closer approximation to the equilibrium state for shorter relaxation delays. The utility of the modified approach is illustrated through simulations on realistic spin systems over a wide range of time scales and experimentally verified using a perdeuterated sample of human ubiquitin. PMID- 18417395 TI - The ontogeny of physiological response to light intensity in early stage spiny lobster (Jasus edwardsii) larvae. AB - Early stage phyllosoma larvae of the spiny rock lobster Jasus edwardsii were examined for swimming speed, feeding, oxygen consumption and nitrogen excretion as instantaneous performance indicators when exposed to different irradiance levels. Swimming speed was measured in recently hatched Stage I larvae while all other parameters were measured in larvae from hatch to mid-Stage V. The swimming speed of recently hatched Stage I phyllosoma increased logarithmically between light intensities of 2.9 x 10(14) and 1.8 x 10(16) quanta s(-1) cm(-2) indicating that, within this range, swimming activity was only suppressed at the lowest irradiance level. Larvae examined under dark (no light) conditions showed lower feed intake, oxygen consumption and nitrogen excretion than larvae under low (7.7 x 10(12) q s(-1) cm(-2)) and high (3.9 x 10(14) q s(-1) cm(-2)) light intensities, and this was a consistent pattern observed throughout development from hatch to Stage V. There was no difference in feeding, oxygen consumption and nitrogen excretion between larvae exposed to low and high light intensities. However, from mid-Stage I to mid-Stage V, the metabolic feeding efficiency (feed intake:oxygen consumption ratio) was consistently higher in larvae exposed to low light intensity than in phyllosoma assessed in the dark and under high irradiance. A light intensity of about 7.7 x 10(12) quanta s(-1) cm(-2) and no higher than 3.9 x 10(14) quanta s(-1) cm(-2) is recommended to stimulate feeding and optimise metabolic feeding efficiency in early larval stages of J. edwardsii. PMID- 18417396 TI - Ontogenesis of catabolic and energy metabolism capacities during the embryonic development of spotted wolffish (Anarhichas minor). AB - The catabolic and energy metabolism capacities during spotted wolffish (Anarhichas minor) embryogenesis were investigated. We assessed the embryo's ability to catabolize proteins (trypsin-like proteases) and lipids (triglyceride lipase) and examined the development of metabolic capacities using enzymatic assays: ability to use carbohydrates (pyruvate kinase), amino acids (aspartate aminotransferase) and fatty acids (hydroxyacyl-CoA dehydrogenase) for energy production, and aerobic (citrate synthase) and anaerobic (lactate dehydrogenase) energy production. Functional enzymatic systems were detected from the eyed stage (350 degree-days), except for fatty acids, which was detected from 540 degree days. To compare the development of 1) aerobic and anaerobic pathways and 2) the capacity to mobilize the different energy substrates, enzymatic ratios were calculated. Anaerobic capacity appeared to increase at a significantly higher rate than the aerobic capacity. Ratios revealing the relative capacity to use specific energy substrates showed a significantly slower increase during development in the capacity to use carbohydrates than amino acids and fatty acids. The end of embryogenesis was characterized by a significant decrease in the use of carbohydrates for aerobic energy production but an increasing capacity to use amino acids. Egg survival as affected by the variability in metabolic parameters is discussed. PMID- 18417397 TI - Changes in body composition during weight loss in obese client-owned cats: loss of lean tissue mass correlates with overall percentage of weight lost. AB - Obesity is one of the most common medical diseases in cats, but there remains little information on success of weight loss regimes in obese client-owned cats. No information currently exists on body composition changes during weight loss in clinical cases. Twelve obese client-owned cats undertook a weight loss programme incorporating a high-protein low fat diet. Body composition was quantified by dual-energy X-ray absorptiometry, before and after weight loss. Mean (+/-standard deviation) weight loss was 27+/-6.8% of starting weight, and mean rate of weight loss was 0.8+/-0.32% per week. Mean energy allocation during weight loss was 32+/ 7.0 kcal/kg target weight. Mean composition of tissue lost was 86:13:1 (fat:lean:bone mineral). The proportion of lean tissue loss was positively associated with overall percentage of weight loss (simple linear regression, r(2)=44.2%, P=0.026). Conventional weight loss programmes produce safe weight loss, but lean tissue loss is an inevitable consequence in cats that lose significant proportions of their starting body weight. PMID- 18417398 TI - Apparent clinical resolution of pinnal actinic keratoses and squamous cell carcinoma in a cat using topical imiquimod 5% cream. AB - Imiquimod is a topical immune response modifier and stimulator used in humans to treat a number of cutaneous neoplasms. This case report describes a cat with actinic keratoses and squamous cell carcinoma of the pinnae. The pinnal lesions were treated with topical 5% imiquimod three times per week. Treatment was discontinued after 82 days of therapy. Twelve weeks of topical imiquimod application resulted in clinical resolution of the pinnal lesions. Although no post-treatment biopsies were performed, there was no relapse of the pinnal lesions in 5 months of clinical follow-up. Expected side effects were limited to erythema, crusting, alopecia, and mild discomfort at the sites of application during the first 3 weeks of application. These results suggest that topical imiquimod, although unproven, might be a therapeutic option or adjunct to therapy for cats with actinic keratoses and squamous cell carcinoma, especially those cats for whom surgery and radiation therapy are not an option. PMID- 18417399 TI - Analysis of non-hospital antibacterial pharmacotherapy in Poland. AB - OBJECTIVES: The aim of this research was to analyze the antibacterial drug consumption pattern in a 1-million-plus strictly defined population in Poland. We assessed outpatient antibiotic sales (ATC J01) in relation to patient age and season of the year, and sought to determine the group of patients with the most frequent recurrence of bacterial diseases. METHODS: The Lubuskie Regional Unit of the National Health Fund (NHF) and the Central Statistical Office (GUS) were the main sources of data. For the period 2002-2005, data on outpatient sales of antibiotics (ATC J01) in Lubuskie Province were collected and expressed in DDD (defined daily dose; World Health Organization anatomical therapeutic chemical (ATC)/DDD version 2006) per 1000 inhabitants per day (DID). RESULTS: During the period 2002-2005, the average level of antibiotic use amounted to 19.8 DID in the Lubuskie population. During the 3-year period, 64.3% of the population was treated with antibiotics. This value varied for different age groups. Of the patients, 22.7% utilized 62.6% of a 3-year supply of antibiotic therapy in the province; a small 1.9% of the population used 10.0% of the supply. The seasonal variation of antibiotic consumption in different age groups showed a strong tendency to be flatter as the older age groups were analyzed. CONCLUSIONS: The data gained from the payers, i.e., the healthcare system, is a very valuable source of information for pharmacoepidemiological studies. Our study shows that seasonal variation in antibiotic use is strictly linked with the age of patients. Special attention should be paid to the relatively small group of patients that utilizes a significant percentage of the year's supply. We have established an effective way to present recurrence data (a map showing the infectious disease incidence). This could be a very useful tool for comparing antibiotic consumption in different countries. PMID- 18417400 TI - Consensus and controversies in platelet transfusion: trigger for indication, and platelet dose. AB - Platelet transfusion is about to commemorate its 50th year since its introduction in therapeutics. It is then surprising to see, that in spite of reaching this respectful age, we have not been able to definitely establish all the aspects related to its clinical use. Some of these facets are platelet transfusion threshold and the platelet dose to administer. Historically, two different transfusion triggers have been used for prophylactic and therapeutic platelet transfusions. For prophylactic platelet transfusion an increasing body of evidences suggests that a transfusion trigger of 10 x 10(9) per liter is appropriate for most clinical settings. In contrast, evidence for supporting a certain therapeutic transfusion trigger is lacking. Nevertheless, there is consensus that the platelet count should not be allowed to fall below 50 x 10(9) per liter in patients with acute bleeding. Another important aspect still pending of clear definition is the issue of the platelet dose to be transfused. It has been addressed by some small studies but a definite answer to this important clinical issue is, at least so far, still pending. The results of two ongoing trials, one sponsored by NIH through the Clinical Trials Network in Transfusion Medicine and Hemostasis and the other promoted by the BEST Collaborative Group are expected to help us to clearly defining the more effective and efficient way to transfuse platelet concentrates. PMID- 18417401 TI - Use of random versus apheresis platelet concentrates. AB - The respective use of random (RPC) and apheresis (APC) platelet concentrates is highly heterogeneous among countries, ranging from 10 to 98% RPC in countries supposed to provide a similar transfusion service to patients. Moreover, when considering each country in the past 10 years, one can observe that some have changed their policy, switching from a majority of APC to RPC or vice versa. This presentation intends to analyse which factors may impact such decisions. For many years, the only available platelet component was a RPC obtained from whole blood donation by a two centrifugation steps process, the "platelet rich plasma" or PRP method. Since the beginning of the 1970s, APCs became available, with in fact many different techniques leading to many APCs that may not be equivalent. Since the end of the 1980s, a new method of RPC preparation was developed, using the buffy-coat (BC-PC), providing a blood component with highly preserved platelet functions as compared to RPCs prepared by the PRP technique. Finally, the use of each of these components either native, or leuco-reduced, or suspended in a storage solution, or processed with a pathogen inactivation technique adds new layers of complexity to compare them. Innumerable references can be found in the literature describing in vitro functional parameters of platelet concentrates. Although it is clear that BC-RPC retain much more their in vitro functions than PRP-RPC, indicating that no one should use the latter any more, it is much more difficult to distinguish differences between other PCs. Conversely, only a very few studies have been published related to a comparison of clinical efficacy of RPC versus APC, the endpoints being mainly CCI. Similarly to the in vitro studies, although RPC prepared with the PRP method show the lowest CCIs, no clear difference exists between "modern" RPC and APC. Another factor that may impact policy decision is the occurrence of adverse reactions in recipients. When considering only comparable data, for example leuco-reduced RPC versus leuco reduced APC, there is now evidence that the latter is more associated with adverse reactions in recipients: data from hemovigilance in France show that, although no difference is noted for febrile non haemolytic transfusion reactions, nor for bacteria contamination, the incidence of allergic adverse reactions is about four times higher with APC as compared with RPC. Other aspects may impact the decision: the fact that using APC in place of RPC reduces the total donor exposure of patients was considered critical in some countries to reduce the risk of transmission of blood transmissible disease. Finally, the cost of the components, much higher for APC may be considered. PMID- 18417402 TI - Uses of plasma in Spain. AB - In Spain, fresh frozen plasma (FFP) currently recovered either by whole blood centrifugation or by apheresis is mainly considered as a source of plasma derivates rather than a product to be transfused. Upon this consideration, the amount of plasma transfused in the last two decades has remained stable, while the production of FFP has grown steadily during all these years. Thus, much more plasma has been derived to industry for manufacturing. Although, since 1993 a consensus conference established the clinical situation where plasma has demonstrated its efficacy, the true situation is that many indications seem not to be supported on a scientific evidence basis. Only a few studies have been performed in the last years to assess the appropriateness of these indications. We present the initial result of an ongoing survey addressed by the Madrid Blood Transfusion Centre. Based on the criteria of total amount of RBC transfused per year, large hospitals (more than 10,000 units of RBC) transfused an average of 23.87% of FFP, while medium hospitals (5000-10,000 units of RBC) used 19.5% and small ones (less than 5000) about 12.5%. It is important to point out that inside each group there were some important differences in ratio values for similar hospitals. This could indicate that much more is necessary to cope with indications. Although national figures of uses of FFP, whether in ratio or absolute terms, show a moderate consumption in comparison with published figures of other European countries, there can be no doubt that plasma overuses still seem to be present. PMID- 18417403 TI - Arabinosylated lipoarabinomannan modulates the impaired cell mediated immune response in Mycobacterium tuberculosis H37Rv infected C57BL/6 mice. AB - Mycobacterium tuberculosis is a facultative intracellular pathogen that flourishes inside the host macrophages. This organism has the ability to deactivate the cell-mediated immune responses involving the down-regulation of pro-inflammatory cytokines, T cell proliferation, apoptosis of CD4+T cells and impairment of the expression of MHC Class II molecules. We observed that Arabinosylated Lipoarabinomannan (Ara-LAM), a glycolipid present in the cell wall of the avirulent Mycobacterium smegmatis, could effectively restrict the growth of tubercle bacilli, induced the transcription of Th1 cytokines in alveolar macrophages (AMs) and splenocytes, enhanced the frequency of CD4+T cells secreting IFN-gamma and induced the expression of MHC Class II molecules on the splenocyte membrane, compared to that of Mycobacterium tuberculosis H37Rv infected C57BL/6 mice. Collectively our findings strongly suggest that Ara-LAM had the potency to restore the impaired cell mediated immune responses in mice infected with Mycobacterium tuberculosis H37Rv, and hence could be utilized as an effective immuno-prophylactic tool in the control of tuberculosis. PMID- 18417404 TI - Autoerythrocyte sensitization syndrome (Gardner-Diamond syndrome) associated with cutaneous vasculitis. AB - Autoerythrocyte sensitization syndrome (ASS) (Gardner-Diamond syndrome) is characterized by painful ecchymotic lesions affecting mostly women with emotional stress. Although it is widely accepted as a non-inflammatory disease, ASS can be accompanied by some autoimmune diseases. In this case report, we present a case with ASS associated with cutaneous vasculitis. We also briefly discuss the possible inflammatory features of ASS. PMID- 18417406 TI - [Is the bronchial atresia prenatal diagnosis possible?]. AB - Bronchial atresia is a rare congenital malformation of the lung. The main-stem segmental or lobar bronchus fails to construct normally, which can lead to accumulation of mucus within the distal bronchi or lung hyperinflation of the obstructed lobe. The prenatal diagnosis is rare and difficult. We report two cases of fetuses who presented pathological examination of the lung on the ultrasonography, at 22 weeks of gestation, suspect of prenatal bronchial atresia diagnosis. We analysed this malformation through a literature review in order to discuss differential diagnosis to be evoked, as well as appropriate perinatal management. PMID- 18417407 TI - [Clearance, persistence and recurrence of HPV infection]. AB - Cervical HPV infection is a common sexually transmitted infection. Most women are infected shortly after beginning their first relationship, with the highest prevalence seen in women under 25 years of age. Thereafter, prevalence decreases rapidly. HPV infections are usually transient; but several factors increasing persistence were identified as host factors (genetic or acquired as age, immunodepression, oral contraception, smoking) and viral factors (genotype, variants, viral load, integration...). Although it is now widely admitted that a persistent infection with a high-risk HPV type is necessary for the development of high-grade cervical intraepithelial neoplasia and invasive disease, whether persistent HPV infections are characterized by the continuing detection of HPV, or by a state of viral latency during which the virus remains undetectable only to reappear later remains unknown. The distinction between a persistent and transient infection is arbitrary depending from both the time of sampling in relation to the natural history of the infection and the interval between samples. The longitudinal studies show that "recurrent" HPV infections offer no evidence that the recurrent episode is correlated with reemergence of the same strain or another strain of the same genotype (wild or variant), but the sequential detection of other HPV type is common. The studies offer no evidence of competition between HPV types but frequently show an increased risk of acquisition of new HPV types in patients already infected compared with those who are HPV-negative. PMID- 18417408 TI - [How I perform... a laparoscopic transabdominal isthmic cerclage]. PMID- 18417409 TI - [Vaccine prevention in perinatal health care: parents, children and professionals]. AB - Recent legislative texts have changed vaccinal policy and reinforced the role of midwives in vaccine prevention in perinatal healthcare. Quite as paediatricians and obstetricians-gynecologists, midwives can now prescribe and carry out, for the mothers, vaccines against rubella, tetanus, poliomyelitis, diphtheria, hepatitis B, influenza and whooping-cough and for the newborns vaccines against hepatitis B and tuberculosis. Concerning vaccinations, practitioners have to respect the vaccination calendar and a collaborative action is useful and necessary. These national guidelines are regularly updated when new vaccines and new recommendations come to light, for example for children (papillomavirus, tuberculosis, pneumococcus...), young adults (varicella, whooping-cough) and health professions in contact with very young children (varicella, measles, influenza and whooping-cough). The recent changes in tuberculosis prevention from routine vaccination of all newborn infants to selective vaccination lead to reinforce measures to detect the infants at higher risk, for them to be vaccinated before discharge at home. Midwives and nurses occupy a central place in family policy and become, with obstetricians-gynecologists and pediatricians, key actors for the effectiveness and the success of vaccine strategies in perinatal health. PMID- 18417410 TI - [Stress urinary incontinence and suburetral slings. Implications for sexuality]. AB - The impact of stress urinary incontinence (SUI) is not limited to physical and psychological consequences classically evaluated for this disease. In fact, some studies emphasize the indisputable existence of sexual disorders directly imputable to SUI. Sexual function is an important evaluation element before and after surgical treatment of SUI. Many validated questionnaires about sexual disorders exist, the most frequently utilized in present literature being the female sexual function index (FSFI). Suburethral slings, in spite of discordant results, do not seem to impair global sexuality of patients, some authors reporting a benefit effect of surgery, related to a decrease of coital incontinence after surgery and a decrease of apprehension before sexual intercourse. PMID- 18417411 TI - Effects of severity of degeneration on gait patterns in patients with medial knee osteoarthritis. AB - This study tested the hypothesis that patients with mild and severe medial knee osteoarthritis (OA) adopt different compensatory gait patterns to unload the deseased knee, in not only the frontal plane but also the sagittal plane. Fifteen patients with mild and 15 with severe bilateral medial knee OA, and 15 normal controls walked while the kinematic and kinetic data were measured. Compared to the normal group, both OA groups had significantly greater pelvic anterior tilt, swing-pelvis list, smaller standing knee abduction, as well as smaller standing hip flexor and knee extensor moments during stance. The severe group also had greater hip abduction, knee extension and ankle plantarflexion. The mild group successfully reduced the extensor moment and maintained normal abductor moment at the diseased knee mainly through listing and anterior tilting the pelvis. With extra compensatory changes at other joints and increased hip abductor moment, the severe group successfully reduced the knee extensor moment but failed to reduce the abductor moment. These results suggest that, apart from training of the knee muscles, training of the hip muscles and pelvic control are essential in the rehabilitative intervention of patients with knee OA, especially for more severe patients. PMID- 18417412 TI - In vivo three-dimensional kinematics of the normal knee during active extension under unloaded and loaded conditions using single-plane fluoroscopy. AB - Measurement of the changes of the three-dimensional (3D) motion and surface kinematics of the knee under different external loading conditions is essential for the understanding and evaluation of the function of the joint, as well as for relevant clinical applications. Knee extension exercise (KEE) has been applied extensively in the rehabilitation programs of patients with various knee disorders. This study measured the 3D knee kinematics of eight normal subjects during active knee extension for unloaded and loaded conditions using a voxel based method for the registration of fluoroscopic images with CT bone data. The knee kinematics during unloaded conditions were found to be similar to previous findings. A mass of 5 kg at the ankle did not affect the joint angles but significantly altered the lateral contact positions during knee extension, especially at knee flexion angles higher than 75 degrees, and also reduced the asymmetry of the surface kinematics between the medial and lateral condyles. The results of the current study may be useful for knee replacement design and for developing guidelines for the use of KEEs for the rehabilitation of patients with knee disorders. PMID- 18417414 TI - A third place in the everyday lives of people living with cancer: functions of Gilda's Club of Greater Toronto. AB - The purpose of this paper was to examine the therapeutic functions of Gilda's Club of Greater Toronto in the everyday lives of people living with cancer. Gilda's Club is a non-institutional setting, where people living with cancer join together to build physical, social, and emotional support as a supplement to medical care. Findings reveal members regarded Gilda's Club as an escape from the stressors of home and hospital, a place where they could meet others living with cancer, and a social environment in which they could confront or distance themselves from their health problems. The paper demonstrates the significance of "third places" for health and calls on researchers to afford such places greater attention. PMID- 18417413 TI - A medical simulator for subcutaneous contraceptive implant insertion. AB - New contraceptive methods like the subcutaneous implant offers a new kind of comfort for women with an efficiency similar to the contraceptive pill. Unfortunately the few numbers of unintended pregnancies that have been reported are generally due to a bad insertion of the implant. In order to give more security to patients, we have designed, in close collaboration with physicians, a new kind of medical simulator. This paper focuses on a device dedicated to a specific subcutaneous implant but it is worth noting that this simulator is relatively generic since it will be used for other subcutaneous techniques or other implant instruments. This simulator can be used for two purposes: one for training novice physicians in the correct manipulation and the other for physician certification which will help determine if they are capable of inserting the implant in vivo. This paper describes the approach which has led to the design of this simulator. It describes its functionalities, its several components but also methods used to analyze the manipulation of the implant insertion inside the patient. Finally first experimental results are reported and discussed. The system used in this paper makes possible to carry out training in a constraint-free context and provides the first mean of visualizing a maneuver that, until now, has been performed blindly. PMID- 18417415 TI - Application of the forward linear prediction on high-resolution NMR spectra in inhomogeneous fields. AB - In homogeneous fields, the advantages of forward linear prediction (LP) for processing 2D NMR data sets have long been recognized. In this paper, the forward LP method was employed to obtain high-resolution NMR spectra in inhomogeneous fields. Intermolecular multiple-quantum coherence (iMQC) signals are caused by intermolecular dipolar interactions and can be used to obtain 1D high-resolution NMR spectra from the 2D iMQC spectra acquired in inhomogeneous fields. However, when the 2D spectra are acquired with insufficient increments to save experimental time, wiggles around strong peaks and bad resolution will occur. Extending the data set by forward LP in the indirect dimension is a good way to improve spectral resolution. Compared to normal discrete Fourier transform, the forward LP method can shorten experimental time by a factor of four or more at the same level of sensitivity and resolution. PMID- 18417416 TI - Analysis of vibrational spectra of 3-halo-1-propanols CH(2)XCH(2)CH(2)OH (X is Cl and Br). AB - The conformational stability and the three rotor internal rotations in 3-chloro- and 3-bromo-1-propanols were investigated by DFT-B3LYP/6-311+G and ab initio MP2/6-311+G, MP3/6-311+G and MP4(SDTQ)//MP3/6-311+G levels of theory. On the calculated potential energy surface twelve distinct minima were located all of which were not predicted to have imaginary frequencies at the B3LYP level of theory. The calculated lowest energy minimum in the potential curves of both molecules was predicted to correspond to the Gauche-gauche-trans (Ggt) conformer in excellent agreement with earlier microwave and electron diffraction results. The equilibrium constants for the conformational interconversion of the two 3 halo-1-propanols were calculated at the B3LYP/6-311+G level of calculation and found to correspond to an equilibrium mixture of about 32% Ggt, 18% Ggg1, 13% Tgt, 8% Tgg and 8% Gtt conformations for 3-chloro-1-propanol and 34% Ggt, 15% Tgt, 13% Ggg1, 9% Tgg and 7% Gtt conformations for 3-bromo-1-propanol at 298.15K. The nature of the high energy conformations was verified by carrying out solvent experiments using formamide ( epsilon=109.5) and MP3 and MP4//MP3 calculations. The vibrational frequencies of each molecule in its three most stable forms were computed at the B3LYP level and complete vibrational assignments were made based on normal coordinate calculations and comparison with experimental data of the molecules. PMID- 18417417 TI - Computer-generated tailored feedback letters for smoking cessation: theoretical and empirical variability of tailoring. AB - PURPOSE: This study examines tailored feedback letters of a smoking cessation intervention that is conceptually based on the transtheoretical model, from a content-based perspective. METHODS: Data of 2 population-based intervention studies, both randomized controlled trials, with total N=1044 were used. The procedure of the intervention, the tailoring principle for the feedback letters, and the content of the intervention materials are described in detail. Theoretical and empirical frequencies of unique feedback letters are presented. RESULTS: The intervention system was able to generate a total of 1040 unique letters with normative feedback only, and almost half a million unique letters with normative and ipsative feedback. Almost every single smoker in contemplation, preparation, action, and maintenance had an empirically unique combination of tailoring variables and received a unique letter. In contrast, many smokers in precontemplation shared a combination of tailoring variables and received identical letters. CONCLUSION: The transtheoretical model provides an enormous theoretical and empirical variability of tailoring. However, tailoring for a major subgroup of smokers, i.e. those who do not intend to quit, needs improvement. Conceptual ideas for additional tailoring variables are discussed. PMID- 18417418 TI - Post-movement beta synchronization in subjects presenting with sensory deafferentation. AB - OBJECTIVE: We studied the time course and location of post-movement beta synchronization (PMBS) in patients presenting with sensory deafferentation, in order to assess the hypothetical relationship between the PMBS and the cortical processing of movement-related somatosensory afferent inputs. METHODS: We used the event-related synchronization (ERS) method. EEG activity was recorded (via a 128-electrode system) during brisk, unilateral right and left index finger extension by 10 patients presenting with neuropathic pain related to sensory deafferentation. Intra- and post-movement changes in beta source power were calculated relative to pre-movement baseline activity. We compared the PMBS results for the painful and non-painful body sides. Furthermore, PMBS patterns in patients were compared with those in nine healthy volunteers. RESULTS: PMBS pattern related to the painful side had a spatial distribution, with an ipsilateral preponderance, significantly more restricted than PMBS pattern on the non-painful side and in the control group. There were no significant differences between patient PMBS patterns on the non-painful side and those in the control group. CONCLUSIONS: Sensory deafferentation disrupts normal PMBS patterns. SIGNIFICANCE: This work provides additional arguments to the hypothesis supporting that the PMBS is influenced by movement-related somatosensory input processing. PMID- 18417419 TI - Sleep phenotypes of intellectual disability: a polysomnographic evaluation in subjects with Down syndrome and Fragile-X syndrome. AB - OBJECTIVE: To analyze sleep architecture and NREM sleep alterations by means of the Cyclic Alternating Pattern (CAP) in children with Down syndrome (DS) and Fragile-X syndrome (fraX), the two most common causes of inherited mental retardation, in order to find out eventual alterations of their sleep microstructure related to their mental retardation phenotypes. METHODS: Fourteen patients affected by fraX (mean age 13.1 years) and 9 affected by Down syndrome (mean age 13.8 years) and 26 age-matched normal controls were included. All subjects underwent overnight polysomnography in the sleep laboratory, after one adaptation night and their sleep architecture and CAP were visually scored. RESULTS: FraX subjects showed a reduced time in bed compared to DS subjects, whereas DS subjects showed a lower sleep efficiency, a higher percentage of wakefulness after sleep onset, and a reduced percentage of stage 2 NREM compared to the other groups. Furthermore, DS and fraX subjects, compared to normal controls, showed a higher percentage of stage 1 NREM and a lower percentage of REM sleep. FraX subjects showed the most disrupted sleep microstructure with low total CAP rate and CAP rate in S2 NREM. Both patient groups showed a lower percentage of A1 and higher percentage of A2 and A3 compared to normal controls. CONCLUSIONS: The analysis of CAP might be able to disclose new important findings in the sleep architecture of children with mental retardation and might characterize sleep microstructural patterns of the different phenotypes of intellectual disability. SIGNIFICANCE: The NREM sleep microstructure alterations found in our subjects, associated with the reduction in REM sleep percentage, seem to be distinctive features of intellectual disability. PMID- 18417420 TI - Skeletal muscle as an immunogenic organ. AB - During the past few years, a possible link between skeletal muscle contractile activity and immune changes has been established. This concept is based on the finding that exercise provokes an increase in a number of cytokines. We have suggested that cytokines and other peptides that are produced; expressed and released by muscle fibers and exert either paracrine or endocrine effects should be classified as 'myokines'. Human skeletal muscle has the capacity to express several myokines belonging to distinct different cytokine classes and contractile activity plays a role in regulating the expression of cytokines in skeletal muscle. In the present review, we focus on the myokines interleukin (IL)-6, IL-8 and IL-15 and their possible anti-inflammatory, immunoregulatory and metabolic roles. PMID- 18417421 TI - Take time to facilitate self-directed learning. PMID- 18417422 TI - Widespread unilateral pain associated with herpes simplex virus infections. AB - We describe a patient group with unexplained widespread pain on one side of the body and pain exacerbations during active labial or genital herpes and during herpetic central nervous system infections. The patients had no visible lesion of the central nervous system on magnetic resonance imaging or abnormality in electrophysiological studies. To understand the nature of the pain and its possible relation to herpes simplex virus (HSV) infections, a clinical neurological examination was performed and quantitative sensory testing and skin biopsies were assessed in 17 patients. The levels of serum total immunoglobulins and IgG subclasses and the frequencies of the immune response genes at the IGH@, HLA-A, -B, -DRB1, C4A, and C4B loci were analyzed in the patients and in control subjects. The patients manifested a uniform clinical syndrome with unilateral pain that was best described as neuropathic and that was exacerbated by HSV reactivations. Low plasma IgG3 concentrations, the presence of either low plasma IgG1 or IgG3 or both, and high anti-HSV-2-IgG titers were more common in the patients than in the control subjects, which rendered the patients more vulnerable to HSV recurrences. PERSPECTIVE: We suggest that low immunoglobulin subclass levels and certain MHC alleles render the patients susceptible to recurring HSV infections. HSV reactivations and the accompanying inflammatory process cause dysfunction of the central nervous system that manifests as neuropathic pain. Studies using functional brain imaging are needed to clarify this syndrome. PMID- 18417424 TI - TIRAP (MAL) S180L polymorphism is a common protective factor against developing tuberculosis and systemic lupus erythematosus. AB - BACKGROUND AND AIM: The involvement of Toll-like receptor (TLR)-mediated pathways in infectious and autoimmunity has been suggested. The MyD88 adaptor-like (Mal) protein, also known as the TIR domain-containing adaptor protein (TIRAP), is implicated in the TLR2- and TLR4-mediated MyD88-dependent signaling pathway. The aim of this study was to investigate the influence of the functional TIRAP (MAL) S180L polymorphism on tuberculosis (TB) and four autoimmune diseases namely: rheumatoid arthritis (RA), primary Sjogren's syndrome (pSS), systemic lupus erythematosus (SLE) and type 1 diabetes mellitus (T1D). METHODS: This was a case control and family based association study in which 1325 individuals from a well defined Colombian population were involved. TIRAP (MAL) S180L genotyping was done by using a polymerase chain reaction-restriction fragment length polymorphism technique and by direct sequencing. RESULTS: Leu180 allele was found to be a protective factor against developing TB (odd ratio (OR): 0.53, 95% confidence interval (CI): 0.29-0.97) and SLE (OR: 0.29, 95% CI: 0.14-0.61) while no significant influence on RA, pSS and T1D was observed. CONCLUSION: These results support the influence of TIRAP (MAL) S180L polymorphism on TB and indicate that TB and SLE might share a common immunogenetic pathway in the innate immune response. PMID- 18417423 TI - The neurobiology of social attachment: A comparative approach to behavioral, neuroanatomical, and neurochemical studies. AB - The formation and maintenance of social bonds in adulthood is an essential component of human health. However studies investigating the underlying neurobiology of such behaviors have been scarce. Microtine rodents offer a unique comparative animal model to explore the neural processes responsible for pair bonding and its associated behaviors. Studies using monogamous prairie voles and other related species have recently offered insight into the neuroanatomical, neurobiological, and neurochemical underpinnings of social attachment. In this review, we will discuss the utility of the microtine rodents in comparative studies by exploring their natural history and social behavior in the laboratory. We will then summarize the data implicating vasopressin, oxytocin, and dopamine in the regulation of pair bonding. Finally, we will discuss the ways in which these neurochemical systems may interact to mediate this complex behavior. PMID- 18417425 TI - Arenal, a new Leptospira serovar of serogroup Javanica, isolated from a patient in Costa Rica. AB - Leptospirosis is a worldwide distributed zoonotic disease caused by pathogenic spirochetes of the genus Leptospira. The basic taxon of Leptospira is the serovar. Currently, nearly 300 serovars have been identified. Leptospirosis is particularly prevalent in warm and humid tropical regions where conditions for transmission and survival of pathogenic leptospires in the environment are optimal. Leptospirosis probably constitutes a serious veterinary and public health problem in Central America but solid figures are missing. To determine distribution of leptospirosis in Costa Rica and to identify locally circulating pathogenic serovars, we performed a sentinel-based study, isolating and characterizing leptospires from patients attending hospitals. Strain MAVJ 401 was isolated from a hospitalized patient in the Alajuela province. The isolate produced agglutination titers notably with reference rabbit antisera against serovars of serogroup Javanica but appeared serologically unique in the standard Cross Agglutinin Absorption Test. Therefore, MAVJ 401 was considered to represent a new serovar, designated Arenal, of the serogroup Javanica. Genotypic analysis revealed that strain MAVJ 401 belongs to Leptospira santarosai, a species that almost exclusively occurs in Latin America. This is not a unique finding of an exotic serovar. Recent isolates from severely ill patients in the same region appeared to be identical to Arenal. We have identified a novel highly virulent serovar from a patient in Costa Rica that is common in this area, thus posing a threat for the local public and veterinary health. PMID- 18417426 TI - Simultaneous determination of dopamine, ascorbic acid and uric acid at poly (Evans Blue) modified glassy carbon electrode. AB - A sensitive and selective electrochemical method for the determination of dopamine using an Evans Blue polymer film modified on glassy carbon electrode was developed. The Evans blue polymer film modified electrode shows excellent electrocatalytic activity toward the oxidation of dopamine in phosphate buffer solution (pH 4.5). The linear range of 1.0 x 10(-6)-3.0 x 10(-5) M and detection limit of 2.5 x 10(-7) M were observed in pH 4.5 phosphate buffer solutions. The interference studies showed that the modified electrode exhibits excellent selectivity in the presence of large excess of ascorbic acid and uric acid. The separation of the oxidation peak potentials for dopamine-ascorbic acid and dopamine-uric acid were about 182 mV and 180 mV, respectively. The differences are large enough to determine AA, DA and UA individually and simultaneously. This work provides a simple and easy approach to selectively detect dopamine in the presence of ascorbic acid and uric acid in physiological samples. PMID- 18417427 TI - Antioxidant treatment reverses mitochondrial dysfunction in a sepsis animal model. AB - Evidence from the literature has demonstrated that reactive oxygen species (ROS) play an important role in the development of multiple organ failure and septic shock. In addition, mitochondrial dysfunction has been implicated in the pathogenesis of multiple organ dysfunction syndrome (MODS). The hypothesis of cytopathic hypoxia postulates that impairment in mitochondrial oxidative phosphorylation reduces aerobic adenosine triphosphate (ATP) production and potentially induces MODS. In this work, our aim was to evaluate the effects of antioxidants on oxidative damage and energy metabolism parameters in liver of rats submitted to a cecal ligation puncture (CLP) model of sepsis. We speculate that CLP induces a sequence of events that culminate with liver cells death. We propose that mitochondrial superoxide production induces mitochondrial oxidative damage, leading to mitochondrial dysfunction, swelling and release of cytochrome c. These events occur in early sepsis development, as reported in the present work. Liver cells necrosis only occurs 24 h after CLP, but all other events occur earlier (6-12 h). Moreover, we showed that antioxidants may prevent oxidative damage and mitochondrial dysfunction in liver of rats after CLP. In another set of experiments, we verified that L-NAME administration did not reverse increase of superoxide anion production, TBARS formation, protein carbonylation, mitochondrial swelling, increased serum AST or inhibition on complex IV activity caused by CLP. Considering that this drug inhibits nitric oxide synthase and that no parameter was reversed by its administration, we suggest that all the events reported in this study are not mediated by nitric oxide. In conclusion, although it is difficult to extrapolate our findings to human, it is tempting to speculate that antioxidants may be used in the future in the treatment of this disease. PMID- 18417428 TI - Simultaneous determination of amitraz and its metabolite in human serum by monolithic silica spin column extraction and liquid chromatography-mass spectrometry. AB - A simple, rapid, sensitive, and specific liquid chromatography-mass spectrometry (LC-MS) method was developed and validated for the quantification of amitraz and its metabolite in human serum. Both the compounds were extracted using monolithic silica spin columns with acetonitrile. The chromatographic separation was performed on a reverse-phase C(18) column with a mobile phase of 10 mM ammonium formate-acetonitrile. The protonated analyte was quantitated in positive ionization by mass spectrometry. The method was validated over the concentration range of 25-1000 ng/ml for amitraz and its metabolite in human serum. For both compounds, the limit of detection was 5 ng/ml. The method was applied to serum samples taken from an attempted suicide patient, and only small volumes of serum were required for the simultaneous determination of these compounds. PMID- 18417429 TI - LC-MS analysis of trimethoxyamphetamine designer drugs (TMA series) from urine samples. AB - A sensitive liquid chromatography-mass spectrometric (LC-MS) method for quantification of an active psychedelic hallucinogenic drugs (trimethoxyamphetamines) in human urine after solid-phase extraction (SPE) with C(18) cartridge was developed and validated. Chromatographic separation was achieved on reversed-phase Phenomenex 3.0 microm Polar Plus column (150 mm x 2.1 mm) with acetonitrile -0.2% acetic acid as mobile-phase and the step gradient elution resulted in a total run time of about 20 min. The analytes were detected by using an electrospray positive ionization mass spectrometry in selected ion monitoring (SIM) mode. In the evaluated concentration range (10-200 ng/mL) (R(2) > or = 0.998) a good linear relationship was obtained. The lower limits of detection (LLODs) and quantification (LLOQs) ranged from 4.26 to 9.12 ng/mL and from 13.18 to 29.22 ng/mL, respectively. Average recoveries ranged from 68.52 to 97.90% in urine at the concentrations of 25, 50 and 100 ng/mL. Intra- and inter day relative standard deviations were 3.70-10.77% and 7.63-12.94%, respectively. This LC-MS method proved to be robust and reliable, and suitable for the use as a confirmation method in clinical urine drug testing. PMID- 18417430 TI - Quantitative determination of besifloxacin, a novel fluoroquinolone antimicrobial agent, in human tears by liquid chromatography-tandem mass spectrometry. AB - A rapid and sensitive method was developed using high-performance liquid chromatography coupled with tandem mass spectrometry (LC/MS/MS) for the quantification of besifloxacin in human tears using sparfloxacin as the internal standard (IS). Besifloxacin was extracted from human tear samples using an ammonium formate buffer at pH 3.25. The method was validated over a concentration range of 2-2000 ng/mL, with a total run time of less than 4 min. The overall intra- and inter-day precision for this method was less than 6%. The method was used to measure besifloxacin concentrations in tear samples collected after topical ocular administration to humans; besifloxacin concentrations were 610+/ 540 microg/g (15 min) and 1.60+/-2.28 microg/g (24h). PMID- 18417431 TI - Rapid and simultaneous determination of tacrolimus (FK506) and diltiazem in human whole blood by liquid chromatography-tandem mass spectrometry: application to a clinical drug-drug interaction study. AB - Tacrolimus (FK506) is a potent immunosuppressant widely used for organ transplantation patients while diltiazem (DTZ), a calcium-channel inhibitor, is often used in renal transplantation patients to prevent post-transplant hypertension. However, DTZ has a significant pharmacokinetic interaction with FK506. In this study, a rapid and sensitive ammonium-adduct based liquid chromatography-tandem mass spectrometry (LC/MS/MS) method has been developed and validated for the simultaneous determination of FK506 and DTZ in human whole blood using ascomycin as the internal standard (IS). After extraction of the whole blood samples by ethyl acetate, FK506, DTZ and the IS were subjected to LC/MS/MS analysis using electro-spray positive-ion mode ionization (ESI(+)). Chromatographic separation was performed on a Hypersil BDS C18 column (50 mm x 2.1 mm, i.d., 3 microm). The MS/MS detection was conducted by monitoring the fragmentation of 821.7-->768.9 (m/z) for FK506, 415.5-->310.3 (m/z) for DTZ and 809.8-->757.0 (m/z) for IS. The method had a chromatographic running time of approximately 2 min and linear calibration curves over the concentrations of 0.5 200 ng/mL for FK506 and 2-250 ng/mL for DTZ. The recoveries of liquid-liquid extraction method were 58.3-62.6% for FK506 and 50.4-58.8% for DTZ. The lower limit of quantification (LLOQ) of the analytical method was 0.5 ng/mL for FK506 and 2 ng/mL for DTZ. The intra- and inter-day precision was less than 15% for all quality control samples at concentrations of 2, 10, and 50 ng/mL for FK506 and 5, 25, and 100 ng/mL for DTZ. The validated LC/MS/MS method has been successfully used to analyze the concentrations of FK506 and DTZ in whole blood samples from pharmacokinetic studies in renal transplanted patients. PMID- 18417432 TI - Quantitative analysis of natural cyclodextrins by high-performance liquid chromatography with pulsed amperometric detection: application to cell permeation study. AB - Simple HPLC-PAD methods were developed for quantitation of cyclodextrins (CDs) in aqueous matrices from in vitro cell permeation studies. C-18 solid-phase extraction was used for sample pretreatment. Samples were analysed using acetonitrile-water mobile phase with post-column alkalization by 0.5M NaOH. Zorbax SB-Aq (for alpha-CD) and Zorbax SB-Phenyl (for beta-CD and gamma-CD) columns gave excellent peak shape and sufficient resolution of CD to glucose (2.7 3.2). The methods showed good concentration-response relationship (r > or = 0.999), precision (RSD% 0.7-5.1), repeatability (RSD% 3.4-13.7) and accuracy (87 107%). The limits of quantitation were 0.78, 0.46 and 0.52 microg/ml for alpha CD, beta-CD and gamma-CD (RSD% of 10.6, 8.1 and 16.3, respectively). PMID- 18417433 TI - Complications of colonoscopy in a large public county hospital in Greece. A 10 year study. AB - BACKGROUND AND STUDY AIMS: Information about the complications of colonoscopy in Southern Europe is limited, particularly in Greece where it is non-existent. Our study sought to determine the complications of colonoscopy in a large public county hospital in Greece over a 10-year period. PATIENTS AND METHODS: All colonoscopy procedures from 1996 to 2006 were entered into a database. Data were analysed by both univariate and multivariate methods. RESULTS: Nine thousand six hundred forty-eight colonoscopies were entered into a database. The procedures were diagnostic in 79% and therapeutic in 21%. Overall bleeding complications occurred in 83 out of the 9648 patients (0.8%: 95% confidence interval [0.7%, 0.9%]). Perforation occurred in four female patients (0.04%: 95% confidence interval [0.01%, 0.07%]) in the sigmoid colon. Multivariate stepwise logistic regression analysis in the therapeutic colonoscopies revealed that presence of significant polyps (odds ratio 4.7, confidence interval [2.9-7.6]), the male sex (odds ratio 2, 95% confidence interval [1.2-3.3]) and the time period of the procedure (the first 5 years) (odds ratio 1.7, 95% confidence interval [1.01-3]), are significant predictors of a post-colonoscopy bleeding episode. CONCLUSION: This historical cohort study, the first in Greece on this subject, shows that colonoscopy is a rather safe procedure and that the rate of complications in this study was low. PMID- 18417434 TI - Prognostic factors of cutaneous melanoma in relation to metastasis at the sentinel lymph node: a case-controlled study. AB - BACKGROUND: The selection of patients who will undergo sentinel lymph node biopsy is primarily based on the histopathologic features of cutaneous melanoma. The purpose of this study is to identify prognostic factors that predict the sentinel lymph node metastasis in melanoma. METHODS: Sixty-nine melanoma patients underwent sentinel lymph node biopsy at Padova Plastic Surgery Institute. Univariate chi-square and multivariate logistic regression analyses were conducted to identify the relationship between prognostic factors and positive sentinel lymph node. A Receiver Operating Characteristics (ROC) Curve was performed to identify the ideal Breslow thickness cutpoint at which to perform sentinel node biopsy. RESULTS: Eleven of the 69 patients (16%) had sentinel lymph node metastases. By univariate analyses Breslow's thickness (p=0.001), ulceration (p=0.001), and lymphovascular invasion (p<0.0001) were found to be significant prognostic factors for the prediction of sentinel node micrometastases. The ROC Curve identified a Breslow thickness of 1.19 mm to be the most suitable cutpoint for sentinel lymph node positivity (p=0.003, sensibility 80%, specificity 69.6%). CONCLUSIONS: Patients with Breslow thickness >or=1.19 mm, ulceration, and lymphovascular invasion are at higher risk for occult lymph node metastases. In addition it is important to use multiple selection criteria when performing sentinel lymph node biopsy especially in patients with thin melanomas. PMID- 18417435 TI - The inconvenience of ear traction through the earring hole in parotidectomy. PMID- 18417436 TI - Surgical correction of axillary accessory breast tissue: 12 cases with emphasis on treatment option. PMID- 18417437 TI - Sex-related differences in recovery of cutaneous nociception after end-to-side nerve repair in the rat. AB - Sex-related differences in the recovery of cutaneous nociception after end-to side nerve repair were examined in rats. Recovery of nociception in the dorsal foot was determined by skin pinch test 19 weeks after the proximal end of the distal stump of the transected peroneal nerve was sutured to the side of the adjacent intact sural nerve (end-to-side nerve coaptation). Axon sprouts in the recipient peroneal nerve were counted by light and electron microscopy. Recovery of nociception due to axon sprouting through the end-to-side coaptation was found in 87% of females and in 60% of males. The area of nociception was not significantly different (P=0.59) between females and males (13+/-8% and 11+/-9%, respectively). The number of myelinated axons in the recipient peroneal nerve (but not of unmyelinated axons) was significantly larger (P=0.028) in females (median=512, 25th and 75th percentiles: 467 and 594) than in males (median=322, 25th and 75th percentiles: 239 and 468). The majority of these axons in females and males were thin fibres, and recipient nerves in both groups were responsive to nerve pinch test. In conclusion, collateral sprouting of thin myelinated nociceptive axons into the end-to-side coapted nerve is more abundant in female than in male rats. However, recovery of cutaneous mechano-nociception due to sprouting of these axons was not different between the two sexes. Possible reasons are discussed. PMID- 18417438 TI - [Acute interstitial nephritis of fluindione: about three cases]. AB - Acute interstitial nephritis is a frequent cause of acute renal failure, representing about 10% of all biopsied cases. Early recognition of drug-induced acute immunoallergic interstitial nephritis prevents the development of severe chronic renal injury. The list of imputable drugs includes phenindione, a vitamin K antagonist. Fluindione which is also an indanedione derivative is another vitamin K antagonist. We report three biopsy-proved cases of fluindione related acute interstitial nephritis with recovery of renal function after drug withdrawal and prednisone therapy. PMID- 18417439 TI - [DOPPS estimate of patient life years attributable to modifiable hemodialysis practices in France]. AB - In this study, we used a prevalent cross-sectional sample of French hemodialysis patients from Dialysis Outcomes and Practice Patterns Study (DOPPS) 2 (2002-2004) to determine the percentage of patients whose values failed to meet targets in six different areas of hemodialysis practice (dialysis dose, anemia, serum phosphorus (PO(4)), serum calcium (Ca), serum albumin and catheter use for vascular access). Cox survival models, with adjustments for patient characteristics, were used for these analyses to estimate mortality hazard ratios (HR). Based on the mortality HR, the fraction of patients outside each target and the total HD population in France, we estimated the number of patient life years that could potentially be gained if every chronic, in-center hemodialysis patient in France who is currently outside of the specified target was able to achieve it. The proportion of patients failing to meet one of the six practice targets in France varied from 15% (dialysis dose) to 75% (albumin) while the percentage of patients complying with all six targets was restricted to 1.2%. The relative risk of mortality (RR) associated with being outside these targets varied from 1.12 to 1.46. Based on these two measures the life-years survival was estimated. The projected number of patients and life years potentially gained from adherence to the six targets was estimated close to 10,600 years-patient. In conclusion, this study suggests large opportunities to improve hemodialysis patient care in France still exist. Compliance with two major practice targets, such as albumin and restriction of catheter use will save highly significant life years of hemodialysis patient. Implementing and strict adherence to national and international guidelines should serve as a basic inspiration for continual improvement of hemodialysis patient care. PMID- 18417441 TI - [Adherence to therapy in sub-Saharan non-dialysed patients with chronic kidney diseases]. AB - BACKGROUND: Poor adherence to medication regimens accounts for substantial morbidity, mortality and increased health care costs in developing countries. The aim of this study is to assess adherence to therapy in patients with chronic kidney diseases and to identify the major barriers to adherence. PATIENTS AND METHOD: A prospective study of non-dialysed patients with chronic kidney diseases followed during three months. Sociodemographic, clinical and therapeutic data were collected from medical records and patient interviews. Rate of adherence (ROA) was defined as the percentage of the prescribed doses of the medication actually taken by the patient over a four-week period. Statistical analysis was done with SPSS 11.0. RESULTS: Mean age of the 118 included patients was 39.28 years +/-16.4 (range 13-76 years); they were 56% female and one third had low instruction level. Socio-economic level was low for 38.5% of patients. Mean ROA was 81+/-12% (range 46 to 100%) but there was a difference between male (75%) and female (84%). Almost three quarters of patients (71%) had ROA more than 80%. Patients' adherence was inversely correlated to daily frequency of dose but not number of drugs. Major obstacles to adherence were complexity of drug regimen, forgetfulness, healthcare system inaccessibility, lack of information, side effects, automedication with phytotherapy and high cost of medications. CONCLUSION: Most of barriers to adherence can be overcome by improving communication between patients, health provider and healthcare system. PMID- 18417442 TI - [First acute rejection episode after renal transplantation: study of the histopathological characteristics according to the immunological risk]. AB - Renal allograft biopsies (n=34) of two different populations of patients according to the immunological risk (high versus low-risk) have been compared retrospectively. The presence of polymorphonuclear leukocytes in peritubular capillaries was more frequent in the high-risk group. The C4d staining was positive in 10% of the low-risk patients and in 50% of the high-risk patients (P=0.03). There were more early graft loss, renal infarctions, interstitial hemorrhage, severe glomerulitis, neutrophilic glomerulitis and Banff III grade rejection in the positive C4d group. In conclusion, half of the immunized patients had a humoral rejection, patients with a C4d positive rejection had more early graft loss and more severe histological lesions. PMID- 18417443 TI - An exploration of factors related to dissemination of and exposure to internet delivered behavior change interventions aimed at adults: a Delphi study approach. AB - BACKGROUND: The Internet is an attractive medium for delivering individualized, computer-tailored behavior change interventions to large numbers of people. However, the actual numbers of people reached seem to fall behind the high expectations. Insight into factors that determine use of and exposure to these Internet interventions is important to be able to increase the reach and improve exposure. OBJECTIVE: The aim was to identify potentially important factors that determine whether adults visit an Internet-delivered behavior change intervention, extend their visit, and revisit the intervention. METHODS: A systematic, three-round Delphi study was conducted among national and international experts from Internet intervention research and practice, e marketing/e-commerce, Web design, and technical website development. In the first round, 30 experts completed a structured, open-ended online questionnaire assessing factors that were, in their opinion, important for a first visit, an extended visit, a revisit and for effective promotion strategies. Based on the responses in this first questionnaire, a closed-ended online questionnaire was developed for use in the second round. A total of 233 experts were invited to complete this questionnaire. Median and interquartile deviation (IQD) scores were computed to calculate agreement and consensus on the importance of the factors. The factors for which no consensus was obtained (IQD > 1) were included in the third-round questionnaire. Factors with a median score of six or higher and with an IQD or =30 or >27 kg/m(2) with treated/untreated hypertension, dyslipidaemia, or both, were randomized to double-blind treatment with placebo, rimonabant 5 or 20 mg once daily plus a calorie-restricted diet for 2 years. Weight loss from baseline to 2 years in the intention-to-treat population was significantly greater with rimonabant 20 mg (mean +/- SD: -5.5 +/- 7.7 kg; P < 0.001) and 5 mg (-2.9 +/- 6.5 kg; P = 0.002) than placebo (-1.2 +/- 6.8 kg). Rimonabant 20 mg produced significantly greater improvements than placebo in waist circumference, high density lipoprotein cholesterol, triglycerides, fasting glucose and insulin levels, insulin resistance, and metabolic syndrome prevalence. Rimonabant 20 mg produced clinically meaningful improvements in all Impact of Weight on Quality of Life-Lite questionnaire domain scores at 2 years. Rimonabant was generally well tolerated and rates of adverse events, including depressed mood disorders and disturbances were similar to placebo during year 2. Proportions of patients with clinically significant depression (Hospital Anxiety and Depression Scale score >11) were similar in all treatment groups. CONCLUSION: Rimonabant 20 mg over 2 years promoted clinically relevant and durable weight loss and improvements in cardiometabolic risk factors. PMID- 18417462 TI - A phase II multicentre study of irinotecan (CPT 11) in combination with cisplatin (CDDP) in metastatic or locally advanced penile carcinoma (EORTC PROTOCOL 30992). AB - BACKGROUND: The aim of this study is to determine efficacy and feasibility of the combination regimen irinotecan and cisplatin in patients with cisplatin advanced penile cancer. PATIENTS AND METHODS: Patients with T3, T4, N1, N2, N3 or M1 cisplatin advanced penile cancer were treated with a combination of irinotecan (60 mg/m(2)) on days 1, 8 and 15 and cisplatin (80 mg/m(2)) administered every 28 days. Patients were treated either in the neo-adjuvant setting for T3 or N1-N2 disease with a maximum of four cycles before surgery or up to eight cycles for T4 or N3 or M1 disease. The study was designed with the aim to exclude a response rate (complete response + partial response) <30% (alpha = 10%, power = 95%). RESULTS: Twenty-eight patients were included and evaluated for toxicity, and 26 eligible patients were evaluated for response. Toxicity was acceptable with three cases of grade 3 diarrhoea and two cases of grade 4 neutropenic fever. There were eight responses (two complete response and six partial response) (30.8%, 80% confidence interval 18.8% to 45.1%): three patients undergoing histological verification after chemotherapy had no evidence of malignancy. CONCLUSION: The study fails to demonstrate a response rate significantly >30%. The observation regarding M0 patients suggests to repeat this study in the neo-adjuvant setting. PMID- 18417463 TI - Literacy skills in children with cochlear implants: the importance of early oral language and joint storybook reading. AB - The goal of this study was to longitudinally examine relationships between early factors (child and mother) that may influence children's phonological awareness and reading skills 3 years later in a group of young children with cochlear implants (N = 16). Mothers and children were videotaped during two storybook interactions, and children's oral language skills were assessed using the "Reynell Developmental Language Scales, third edition." Three years later, phonological awareness, reading skills, and language skills were assessed using the "Phonological Awareness Test," the "Woodcock-Johnson-III Diagnostic Reading Battery," and the "Oral Written Language Scales." Variables included in the data analyses were child (age, age at implant, and language skills) and mother factors (facilitative language techniques) and children's phonological awareness and reading standard scores. Results indicate that children's early expressive oral language skills and mothers' use of a higher level facilitative language technique (open-ended question) during storybook reading, although related, each contributed uniquely to children's literacy skills. Individual analyses revealed that the children with expressive standard scores below 70 at Time 1 also performed below average (<85) on phonological awareness and total reading tasks 3 years later. Guidelines for professionals are provided to support literacy skills in young children with cochlear implants. PMID- 18417464 TI - Seeing the deaf in "deafness". AB - This article draws on some of the existing literature on the politics of identity and representation as related to minority group formation. It applies this to constructions of Deaf2 identity from a cultural and linguistic perspective and contrasts this with dominant constructions of Deaf people as disabled. It highlights a number of ways in which Deaf identity differs from disabled identity, demonstrating that the cultural and linguistic construction of Deaf people is a more useful tool for analysis. It raises questions aimed to examine the discourse on deafness and seeks further debate on how best the discourse can be progressed. The article raises issues related to the use of terminology and labeling in the field of deafness. It contends that the continued use of the word deafness is unworkable and should be more widely recognized as a social construct, which has current usage beyond the paradigm in which it was originally intended. The article concludes by recognizing the importance of diversity in identity formation, while simultaneously calling for an appreciation of the need to incorporate this diversity within wider theorizing, focused on commonality and cohesion in identity as a source of collective expression and political mobilization. PMID- 18417465 TI - Simply no time? Barriers to GPs' participation in primary health care research. AB - BACKGROUND: Non-participation of general practitioners (GPs) is a serious source of bias for practice-based studies. Objective. To elucidate doctors' motives for non-participation in, and subjective barriers to, general practice research. METHODS: German GPs that had opted out of a quality assessment project involving electronic patient records (EPRs) were mailed a questionnaire regarding their attitudes towards general practice research and their specific objections to the current project. A sub-sample of doctors was interviewed. Their statements were coded and classified with regard to the reasons given for non-participation and possible motivating factors. RESULTS: The survey response rate was 37% (96/263); 21 GPs completed an additional qualitative interview. Nearly all respondents (88/96) considered general practice research to be important, but 58% had not previously participated in research projects and 56% would not do so in the future. Nearly half (47/96) were opposed to having data extracted from their EPRs. The qualitative analysis revealed deep concerns related to the collection of EPRs (e.g. potential misuse of data, being subject to control or resulting computer problems). Some GPs expressed concerns about recruiting their own patients for the study. Some doctors complained of not being sufficiently recognized as a partner or not having a voice in the research process. CONCLUSION: Doctors' negative attitudes, concerns and ambivalent feelings should be addressed in recruitment strategies, especially when the analysis of EPRs or direct patient contact is required. Some doctors do not participate in research out of principle and will be very difficult to convince. PMID- 18417466 TI - Antipsychotic combinations vs monotherapy in schizophrenia: a meta-analysis of randomized controlled trials. AB - CONTEXT: Despite lacking evidence for its safety and efficacy, antipsychotic cotreatment is common in schizophrenia. OBJECTIVE: To evaluate therapeutic and adverse effects of antipsychotic cotreatment vs monotherapy in schizophrenia. DATA SOURCES: Cochrane Schizophrenia Group register and hand searches of relevant journals/conference proceedings. STUDY SELECTION: Randomized controlled trials comparing antipsychotic monotherapy to cotreatment with a second antipsychotic. DATA EXTRACTION AND ANALYSIS: Two authors independently extracted data. For homogenous dichotomous data, we calculated random effects, relative risk (RR), 95% confidence intervals (CIs), and numbers needed to treat (NNT). For continuous data, weighted mean differences were calculated. RESULTS: In 19 studies (1229 patients) with 28 monotherapy and 19 cotreatment arms, antipsychotic cotreatment was superior to monotherapy regarding 2 a priori defined coprimary outcomes: less study-specific defined inefficacy (N = 22, n = 1202, RR = 0.76, CI = 0.63-0.90, P = .002, NNT = 7, CI = 4-17, P = .0008, I(2) = 78.9%) and all-cause discontinuation (N = 20, n = 1052, RR = 0.65, CI = 0.54-0.78, P < .00001). Results were consistent using Clinical Global Impressions thresholds of less than much (P = .006) and less than minimally (P = .01) improved. Specific psychopathology and adverse event data were insufficient to yield meaningful results. In sensitivity analyses, 5 efficacy moderators emerged: concurrent polypharmacy initiation, clozapine combinations, trial duration >10 weeks, Chinese trials, and second-generation + first-generation antipsychotics. In a meta-regression, similar dose combinations, second-generation + first-generation antipsychotics and concurrent polypharmacy initiation remained significant. CONCLUSIONS: In certain clinical situations, antipsychotic cotreatment may be superior to monotherapy. However, the database is subject to possible publication bias and too heterogeneous to derive firm clinical recommendations, underscoring the need for future research. PMID- 18417467 TI - Structural basis of proteolytic activation of L-phenylalanine oxidase from Pseudomonas sp. P-501. AB - The mature form of l-phenylalanine oxidase (PAOpt) from Pseudomonas sp. P-501 was generated and activated by the proteolytic cleavage of a noncatalytic proenzyme (proPAO). The crystal structures of proPAO, PAOpt, and the PAOpt-o-amino benzoate (AB) complex were determined at 1.7, 1.25, and 1.35A resolutions, respectively. The structure of proPAO suggests that the prosequence peptide of proPAO occupies the funnel (pathway) of the substrate amino acid from the outside of the protein to the interior flavin ring, whereas the funnel is closed with the hydrophobic residues at its vestibule in both PAOpt and the PAOpt-AB complex. All three structures have an oxygen channel that is open to the surface of the protein from the flavin ring. These results suggest that structural changes were induced by proteolysis; that is, the proteolysis of proPAO removes the prosequence and closes the funnel to keep the active site hydrophobic but keeps the oxygen channel open. The possibility that an interaction of the hydrophobic side chain of substrate with the residues of the vestibule region may open the funnel as a putative amino acid channel is discussed. PMID- 18417468 TI - Solution structure of a hydrocarbon stapled peptide inhibitor in complex with monomeric C-terminal domain of HIV-1 capsid. AB - The human immunodeficiency virus type 1 (HIV-1) capsid protein plays a critical role in virus core particle assembly and is an important target for novel therapeutic strategies. In a previous study, we characterized the binding affinity of a hydrocarbon stapled helical peptide, NYAD-1, for the capsid protein (K(d) approximately 1 mum) and demonstrated its ability to penetrate the cell membrane (Zhang, H., Zhao, Q., Bhattacharya, S., Waheed, A. A., Tong, X., Hong, A., Heck, S., Goger, M., Cowburn, D., Freed, E. O., and Debnath, A. K. (2008) J. Mol. Biol. 378, 565-580). In cell-based assays, NYAD-1 colocalized with the Gag polyprotein during traffic to the plasma membrane and disrupted the formation of mature and immature virus particles in vitro systems. Here, we complement the cellular and biochemical data with structural characterization of the interactions between the capsid and a soluble peptide analogue, NYAD-13. Solution NMR methods were used to determine a high resolution structure of the complex between the inhibitor and a monomeric form of the C-terminal domain of the capsid protein (mCA-CTD). The intermolecular interactions are mediated by the packing of hydrophobic side chains at the buried interface and unperturbed by the presence of the olefinic chain on the solvent-exposed surface of the peptide. The results of the structural analysis provide valuable insight into the determinants for high affinity and selective inhibitors for HIV-1 particle assembly. PMID- 18417469 TI - A dual task for the Xbp1-responsive OS-9 variants in the mammalian endoplasmic reticulum: inhibiting secretion of misfolded protein conformers and enhancing their disposal. AB - Normally, non-native polypeptides are not transported through the secretory pathway. Rather, they are translocated from the endoplasmic reticulum (ER) lumen into the cytosol where they are degraded by proteasomes. Here we characterize the function in ER quality control of two proteins derived from alternative splicing of the OS-9 gene. OS-9.1 and OS-9.2 are ubiquitously expressed in human tissues and are amplified in tumors. They are transcriptionally induced upon activation of the Ire1/Xbp1 ER-stress pathway. OS-9 variants do not associate with folding competent proteins. Rather, they selectively bind folding-defective ones thereby inhibiting transport of non-native conformers through the secretory pathway. The intralumenal level of OS-9.1 and OS-9.2 inversely correlates with the fraction of a folding-defective glycoprotein, the Null(hong kong) (NHK) variant of alpha1 antitrypsin that escapes retention-based ER quality control. OS-9 up-regulation does not affect NHK disposal, but reduction of the intralumenal level of OS-9.1 and OS-9.2 substantially delays disposal of this model substrate. OS-9.1 and OS 9.2 also associate transiently with non-glycosylated folding-defective proteins, but association is unproductive. Finally, OS-9 activity does not require an intact mannose 6-P homology domain. Thus, OS-9.1 and OS-9.2 play a dual role in mammalian ER quality control: first as crucial retention factors for misfolded conformers, and second as promoters of protein disposal from the ER lumen. PMID- 18417470 TI - Re-evaluation of chicken CXCR1 determines the true gene structure: CXCLi1 (K60) and CXCLi2 (CAF/interleukin-8) are ligands for this receptor. AB - The original report of chicken CXCR1 (Li, Q. J., Lu, S., Ye, R. D., and Martins Green, M. (2000) Gene (Amst.) 257, 307-317) described it as a single exon gene, with two isoforms (differing in their start codon). In comparison with mammalian CXCR1, the reported chicken CXCR1 was longer at both the NH(2) and COOH termini, and it lacked the conserved (C/S)CXNP motif present in the last transmembrane region of all known chemokine receptors. A re-evaluation of chicken CXCR1, comparing known expressed sequence tags with the chicken genome sequence, suggested that the gene contains two exons. We isolated a cDNA corresponding to our prediction, which was significantly different in sequence to the reported CXCR1. In particular, there were three frameshifts in our sequence, compared with the reported sequence, that restored higher identity in the COOH-terminal half of the protein to mammalian CXCR1 (61% total amino acid identity compared with 52% for the reported CXCR1), restored the (C/S)CXNP motif, and gave a predicted protein of the same length as mammalian CXCR1. In human, CXCR1 is the receptor for CXCL8. In the chicken, there are two syntenic genes, CXCLi1 and CXCLi2, which look equally like orthologues of human CXCL8. We demonstrate that both of these chemokines are ligands for chicken CXCR1. We also demonstrate that heterophils express chicken CXCR1 and that the receptor is Galpha(i) protein-linked. PMID- 18417471 TI - Impact of phosphorylation and phosphorylation-null mutants on the activity and deamination specificity of activation-induced cytidine deaminase. AB - Activation-induced cytidine deaminase (AID) initiates somatic hypermutation and class switch recombination in B cells by deaminating C --> U on transcribed DNA. Here we analyze the role of phosphorylation and phosphorylation-null mutants on the biochemical behavior of AID, including enzyme specific activity, processivity, deamination spectra, deamination motif specificity, and transcription-dependent deamination in the presence and absence of RPA. We show that a small fraction of recombinant human AID expressed in Sf9 insect cells is phosphorylated at previously identified residues Ser(38) and Thr(27) and also at Ser(41) and Ser(43). S43P AID has been identified in a patient with hyper-IgM immunodeficiency syndrome. Ser-substituted phosphorylation-null mutants (S38A, S41A, S43A, and S43P) exhibit wild type (WT) activity on single-stranded DNA. Deamination of transcribed double-stranded DNA is similar for WT and mutant AID and occurs with or without RPA. Although WT and AID mutants catalyze processive deamination favoring canonical WRC hot spot motifs (where W represents A/T and R is A/G), their deamination spectra differ significantly. The differences between the WT and AID mutants appear to be caused by the replacement of Ser as opposed to an absence of phosphorylation. The spectral differences reflect a marked change in deamination efficiencies in two motifs, GGC and AGC, which are preferred by mutant AID but disfavored by WT AID. Both motifs occur with exceptionally high frequency in human switch regions, suggesting a possible relationship between AID deamination specificity and a loss of antibody diversification. PMID- 18417473 TI - Metabolic interrelations within guanine deoxynucleotide pools for mitochondrial and nuclear DNA maintenance. AB - Mitochondrial deoxynucleoside triphosphates are formed and regulated by a network of anabolic and catabolic enzymes present both in mitochondria and the cytosol. Genetic deficiencies for enzymes of the network cause mitochondrial DNA depletion and disease. We investigate by isotope flow experiments the interrelation between mitochondrial and cytosolic deoxynucleotide pools as well as the contributions of the individual enzymes of the network to their maintenance. To study specifically the synthesis of dGTP used for the synthesis of mitochondrial and nuclear DNA, we labeled hamster CHO cells or human fibroblasts with [(3)H]deoxyguanosine during growth and quiescence and after inhibition with aphidicolin or hydroxyurea. At time intervals we determined the labeling of deoxyguanosine nucleotides and DNA and the turnover of dGTP from its specific radioactivity in the separated mitochondrial and cytosolic pools. In both cycling and quiescent cells, the import of deoxynucleotides formed by cytosolic ribonucleotide reductase accounted for most of the synthesis of mitochondrial dGTP, with minor contributions by cytosolic deoxycytidine kinase and mitochondrial deoxyguanosine kinase. A dynamic isotopic equilibrium arose rapidly from the shuttling of deoxynucleotides between mitochondria and cytosol, incorporation of dGTP into DNA, and degradation of dGMP. Inhibition of DNA synthesis by aphidicolin marginally affected the equilibrium. Inhibition of DNA synthesis by blockage of ribonucleotide reduction with hydroxyurea instead disturbed the equilibrium and led to accumulation of labeled dGTP in the cytosol. The turnover of dGTP decreased, suggesting a close connection between ribonucleotide reduction and pool degradation. PMID- 18417472 TI - Structure of the integrin alphaIIb transmembrane segment. AB - Integrin cell-adhesion receptors transduce signals bidirectionally across the plasma membrane via the single-pass transmembrane segments of each alpha and beta subunit. While the beta3 transmembrane segment consists of a linear 29-residue alpha-helix, the structure of the alphaIIb transmembrane segment reveals a linear 24-residue alpha-helix (Ile-966 -Lys-989) followed by a backbone reversal that packs Phe-992-Phe-993 against the transmembrane helix. The length of the alphaIIb transmembrane helix implies the absence of a significant transmembrane helix tilt in contrast to its partnering beta3 subunit. Sequence alignment shows Gly-991-Phe 993 to be fully conserved among all 18 human integrin alpha subunits, suggesting that their unusual structural motif is prototypical for integrin alpha subunits. The alphaIIb transmembrane structure demonstrates a level of complexity within the membrane that is beyond simple transmembrane helices and forms the structural basis for assessing the extent of structural and topological rearrangements upon alphaIIb-beta3 association, i.e. integrin transmembrane signaling. PMID- 18417474 TI - The M1P1 loop of TASK3 K2P channels apposes the selectivity filter and influences channel function. AB - Channels of the two-pore domain potassium (K2P) family contain two pore domains rather than one and an unusually long pre-pore extracellular linker called the M1P1 loop. The TASK (TASK1, TASK3, and TASK5) subfamily of K2P channels is regulated by a number of different pharmacological and physiological mediators. At pH 7.4 TASK3 channels are selectively blocked by zinc in a manner that is both pH(o)- and [K](o)(-)dependent. Mutation of both the Glu-70 residue in the M1P1 loop and the His-98 residue in the pore region abolished block, suggesting the two residues may contribute to a zinc binding site. Mutation of one Glu-70 residue and one His-98 residue to cysteine in TASK3 fixed concatamer channels gave currents that were enhanced by dithiothreitol and then potently blocked by cadmium, suggesting that spontaneous disulfide bridges could be formed between these two residues. Swapping the M1P1 loops of TASK1 and TASK3 channels showed that the M1P1 loop is also involved in channel regulation by pH. Therefore, the TASK3 M1P1 loop lies close to the pore, regulating TASK3 channel activity. PMID- 18417475 TI - Unique Asn-linked oligosaccharides of the human pathogen Entamoeba histolytica. AB - N-Glycans of Entamoeba histolytica, the protist that causes amebic dysentery and liver abscess, are of great interest for multiple reasons. E. histolytica makes an unusual truncated N-glycan precursor (Man(5)GlcNAc(2)), has few nucleotide sugar transporters, and has a surface that is capped by the lectin concanavalin A. Here, biochemical and mass spectrometric methods were used to examine N-glycan biosynthesis and the final N-glycans of E. histolytica with the following conclusions. Unprocessed Man(5)GlcNAc(2), which is the most abundant E. histolytica N-glycan, is aggregated into caps on the surface of E. histolytica by the N-glycan-specific, anti-retroviral lectin cyanovirin-N. Glc(1)Man(5)GlcNAc(2), which is made by a UDP-Glc: glycoprotein glucosyltransferase that is part of a conserved N-glycan-dependent endoplasmic reticulum quality control system for protein folding, is also present in mature N glycans. A swainsonine-sensitive alpha-mannosidase trims some N-glycans to biantennary Man(3)GlcNAc(2). Complex N-glycans of E. histolytica are made by the addition of alpha1,2-linked Gal to both arms of small oligomannose glycans, and Gal residues are capped by one or more Glc. In summary, E. histolytica N-glycans include unprocessed Man(5)GlcNAc(2), which is a target for cyanovirin-N, as well as unique, complex N-glycans containing Gal and Glc. PMID- 18417476 TI - Osteopontin negatively regulates parathyroid hormone receptor signaling in osteoblasts. AB - Systemic hormonal control exerts its effect through the regulation of local target tissues, which in turn regulate upstream signals in a feedback loop. The parathyroid hormone (PTH) axis is a well defined hormonal signaling system that regulates calcium levels and bone metabolism. To understand the interplay between systemic and local signaling in bone, we examined the effects of deficiency of the bone matrix protein osteopontin (OPN) on the systemic effects of PTH specifically within osteoblastic cell lineages. Parathyroid hormone receptor (PPR) transgenic mice expressing a constitutively active form of the receptor (caPPR) specifically in cells of the osteoblast lineage have a high bone mass phenotype. In these mice, OPN deficiency further increased bone mass. This increase was associated with conversion of the major intertrabecular cell population from hematopoietic cells to stromal/osteoblastic cells and parallel elevations in histomorphometric and biochemical parameters of bone formation and resorption. Treatment with small interfering RNA (siRNA) for osteopontin enhanced H223R mutant caPPR-induced cAMP-response element (CRE) activity levels by about 10-fold. Thus, in addition to the well known calcemic feedback system for PTH, local feedback regulation by the bone matrix protein OPN also plays a significant role in the regulation of PTH actions. PMID- 18417477 TI - The apolipoprotein E-mimetic peptide COG112 inhibits the inflammatory response to Citrobacter rodentium in colonic epithelial cells by preventing NF-kappaB activation. AB - Inflammatory bowel disease arises from the interplay between luminal bacteria and the colonic mucosa. Targeted inhibition of pro-inflammatory pathways without global immunosuppression is highly desirable. Apolipoprotein (apo) E has immunomodulatory effects and synthetically derived apoE-mimetic peptides are beneficial in models of sepsis and neuroinflammation. Citrobacter rodentium is the rodent equivalent of enteropathogenic Escherichia coli, and it causes colitis in mice by colonizing the surface of colonic epithelial cells and inducing signaling events. We have reported that mice deficient in inducible nitric-oxide (NO) synthase (iNOS) have attenuated C. rodentium-induced colitis. We used young adult mouse colon (YAMC) cells that mimic primary colonic epithelial cells to study effects of an antennapedia-linked apoE-mimetic peptide, COG112, on C. rodentium-activated cells. COG112 significantly attenuated induction of NO production, and iNOS mRNA and protein expression, in a concentration-dependent manner. COG112 inhibited the C. rodentium-stimulated induction of iNOS and the CXC chemokines KC and MIP-2 to the same degree as the NF-kappaB inhibitors MG132 or BAY 11-7082, and there was no additive effect when COG112 and these inhibitors were combined. COG112 significantly reduced nuclear translocation of NF-kappaB, when assessed by electromobility shift assay, immunoblotting, and immunofluorescence for p65. This correlated with inhibition of both C. rodentium stimulated IkappaB-alpha phosphorylation and degradation, and IkappaB kinase activity, which occurred by inhibition of IkappaB kinase complex formation rather than by a direct effect on the enzyme itself. These studies indicate that apoE mimetic peptides may have novel therapeutic potential by inhibiting NF-kappaB driven proinflammatory epithelial responses to pathogenic colonic bacteria. PMID- 18417478 TI - Competitive interactions of collagen and a jararhagin-derived disintegrin peptide with the integrin alpha2-I domain. AB - Integrin alpha2beta1 is a major receptor required for activation and adhesion of platelets, through the specific recognition of collagen by the alpha2-I domain (alpha2-I), which binds fibrillar collagen via Mg(2+)-bridged interactions. The crystal structure of a truncated form of the alpha2-I domain, bound to a triple helical collagen peptide, revealed conformational changes suggestive of a mechanism where the ligand-bound I domain can initiate and propagate conformational change to the full integrin complex. Collagen binding by alpha2-I and fibrinogen-dependent platelet activity can be inhibited by snake venom polypeptides. Here we describe the inhibitory effect of a short cyclic peptide derived from the snake toxin metalloprotease jararhagin, with specific amino acid sequence RKKH, on the ability of alpha2-I to bind triple helical collagen. Isothermal titration calorimetry measurements showed that the interactions of alpha2-I with collagen or RKKH peptide have similar affinities, and NMR chemical shift mapping experiments with (15)N-labeled alpha2-I, and unlabeled RKKH peptide, indicate that the peptide competes for the collagen-binding site of alpha2-I but does not induce a large scale conformational rearrangement of the I domain. PMID- 18417479 TI - MicroRNA-210 modulates endothelial cell response to hypoxia and inhibits the receptor tyrosine kinase ligand Ephrin-A3. AB - MicroRNAs (miRNAs) are small non-protein-coding RNAs that function as negative gene expression regulators. In the present study, we investigated miRNAs role in endothelial cell response to hypoxia. We found that the expression of miR-210 progressively increased upon exposure to hypoxia. miR-210 overexpression in normoxic endothelial cells stimulated the formation of capillary-like structures on Matrigel and vascular endothelial growth factor-driven cell migration. Conversely, miR-210 blockade via anti-miRNA transfection inhibited the formation of capillary-like structures stimulated by hypoxia and decreased cell migration in response to vascular endothelial growth factor. miR-210 overexpression did not affect endothelial cell growth in both normoxia and hypoxia. However, anti-miR 210 transfection inhibited cell growth and induced apoptosis, in both normoxia and hypoxia. We determined that one relevant target of miR-210 in hypoxia was Ephrin-A3 since miR-210 was necessary and sufficient to down-modulate its expression. Moreover, luciferase reporter assays showed that Ephrin-A3 was a direct target of miR-210. Ephrin-A3 modulation by miR-210 had significant functional consequences; indeed, the expression of an Ephrin-A3 allele that is not targeted by miR-210 prevented miR-210-mediated stimulation of both tubulogenesis and chemotaxis. We conclude that miR-210 up-regulation is a crucial element of endothelial cell response to hypoxia, affecting cell survival, migration, and differentiation. PMID- 18417481 TI - Lack of DNA mismatch repair protein MSH6 in the rat results in hereditary non polyposis colorectal cancer-like tumorigenesis. AB - To understand genetic instability in relation to tumorigenesis, experimental animal models have proven very useful. The DNA mismatch repair (MMR) machinery safeguards genomic integrity by repairing mismatches, insertion or deletion loops and responding to genotoxic agents. Here, we describe the functional characterization of a novel rat mutant model in which the MMR gene Msh6 has been genetically inactivated by N-ethyl-N-nitrosourea-driven target-selected mutagenesis. This model shows a robust mutator phenotype that is reflected by microsatellite instability and an increased germ line point mutation frequency. Consequently, these rats develop a spectrum of tumors with a high similarity to atypical hereditary non-polyposis colorectal cancer in humans. The MSH6 knockout rat complements existing models for studying genetic instable tumorigenesis as it provides experimental opportunities that are not available or suboptimal in current models. PMID- 18417480 TI - Conformational isomerism can limit antibody catalysis. AB - Ligand binding to enzymes and antibodies is often accompanied by protein conformational changes. Although such structural adjustments may be conducive to enzyme catalysis, much less is known about their effect on reactions promoted by engineered catalytic antibodies. Crystallographic and pre-steady state kinetic analyses of antibody 34E4, which efficiently promotes the conversion of benzisoxazoles to salicylonitriles, show that the resting catalyst adopts two interconverting active-site conformations, only one of which is competent to bind substrate. In the predominant isomer, the indole side chain of Trp(L91) occupies the binding site and blocks ligand access. Slow conformational isomerization of this residue, on the same time scale as catalytic turnover, creates a deep and narrow binding site that can accommodate substrate and promote proton transfer using Glu(H50) as a carboxylate base. Although 34E4 is among the best catalysts for the deprotonation of benzisoxazoles, its efficiency appears to be significantly limited by this conformational plasticity of its active site. Future efforts to improve this antibody might profitably focus on stabilizing the active conformation of the catalyst. Analogous strategies may also be relevant to other engineered proteins that are limited by an unfavorable conformational pre equilibrium. PMID- 18417482 TI - Catalysis and regulation in Rubisco. AB - Ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) catalyses the incorporation of inorganic CO(2) into the organic molecules of life. Rubisco is extremely inefficient as a catalyst and its carboxylase activity is compromised by numerous side-reactions including oxygenation of its sugar phosphate substrate by atmospheric O(2). The reduction in the catalytic efficiency as a result of these processes has implications for crop yield, nitrogen and water usage, and for the global carbon cycle. Several aspects of Rubisco including its complex biosynthesis and multi-step catalytic reaction are subject to tight control involving light, cellular metabolites, and molecular chaperones. Numerous high resolution crystal structures of different forms of Rubisco are now available, including structures of mutant enzymes. These provide a molecular framework for the understanding of these processes at the molecular level. PMID- 18417483 TI - Genetic or pharmacologic amplification of nrf2 signaling inhibits acute inflammatory liver injury in mice. AB - Oxidative stress-mediated destruction of normal parenchymal cells during hepatic inflammatory responses contributes to the pathogenesis of immune-mediated hepatitis and is implicated in the progression of acute inflammatory liver injury to chronic inflammatory liver disease. The transcription factor NF-E2-related factor 2 (Nrf2) regulates the expression of a battery of antioxidative enzymes and Nrf2 signaling can be activated by small-molecule drugs that disrupt Keap1 mediated repression of Nrf2 signaling. Therefore, genetic and pharmacologic approaches were used to activate Nrf2 signaling to assess protection against inflammatory liver injury. Profound increases in indicators of cell death were observed in both Nrf2 wild-type (Nrf2-WT) mice and Nrf2-disrupted (Nrf2-KO) mice 24 h following intravenous injection of concanavalin A (12.5 mg/kg, ConA), a model for T cell-mediated acute inflammatory liver injury. However, hepatocyte specific conditional Keap1 null (Alb-Cre:Keap1(flox/-), cKeap1-KO) mice with constitutively enhanced expression of Nrf2-regulated antioxidative genes as well as Nrf2-WT mice but not Nrf2-KO mice pretreated with three daily doses of a triterpenoid that potently activates Nrf2 (30 mumol/kg, cyano-3,12-dioxooleana 1,9(11)-dien-28-oyl-imidazolide [CDDO-Im]) were highly resistant to ConA-mediated inflammatory liver injury. CDDO-Im pretreatment of both Nrf2-WT and Nrf2-KO mice resulted in equivalent suppression of serum proinflammatory soluble proteins suggesting that the hepatoprotection afforded by CDDO-Im pretreatment of Nrf2-WT mice but not Nrf2-KO mice was not due to suppression of systemic proinflammatory signaling, but instead was due to activation of Nrf2 signaling in the liver. Enhanced hepatic expression of Nrf2-regulated antioxidative genes inhibited inflammation-mediated oxidative stress, thereby preventing hepatocyte necrosis. Attenuation of hepatocyte death in cKeap1-KO mice and CDDO-Im pretreated Nrf2-WT mice resulted in decreased late-phase proinflammatory gene expression in the liver thereby diminishing the sustained influx of inflammatory cells initially stimulated by the ConA challenge. Taken together, these results clearly illustrate that targeted cytoprotection of hepatocytes through Nrf2 signaling during inflammation prevents the amplification of inflammatory responses in the liver. PMID- 18417484 TI - High rates of molecular evolution in hantaviruses. AB - Hantaviruses are rodent-borne Bunyaviruses that infect the Arvicolinae, Murinae, and Sigmodontinae subfamilies of Muridae. The rate of molecular evolution in the hantaviruses has been previously estimated at approximately 10(-7) nucleotide substitutions per site, per year (substitutions/site/year), based on the assumption of codivergence and hence shared divergence times with their rodent hosts. If substantiated, this would make the hantaviruses among the slowest evolving of all RNA viruses. However, as hantaviruses replicate with an RNA dependent RNA polymerase, with error rates in the region of one mutation per genome replication, this low rate of nucleotide substitution is anomalous. Here, we use a Bayesian coalescent approach to estimate the rate of nucleotide substitution from serially sampled gene sequence data for hantaviruses known to infect each of the 3 rodent subfamilies: Araraquara virus (Sigmodontinae), Dobrava virus (Murinae), Puumala virus (Arvicolinae), and Tula virus (Arvicolinae). Our results reveal that hantaviruses exhibit short-term substitution rates of 10(-2) to 10(-4) substitutions/site/year and so are within the range exhibited by other RNA viruses. The disparity between this substitution rate and that estimated assuming rodent-hantavirus codivergence suggests that the codivergence hypothesis may need to be reevaluated. PMID- 18417485 TI - Polyploid speciation did not confer instant reproductive isolation in Capsella (Brassicaceae). AB - Polyploid formation is a major mode of sympatric speciation in flowering plants. Unlike other speciation processes, polyploidization is often assumed to confer instant reproductive isolation. Shared polymorphism across ploidy levels has therefore often been attributed to multiple polyploid origins, whereas the alternative hypothesis of introgressive hybridization has rarely been rigorously tested. Here, we sequence 12 nuclear loci representing 6 genes duplicated by polyploidy in 92 accessions of the tetraploid Capsella bursa-pastoris together with the corresponding loci in 21 accessions of its close diploid relative Capsella rubella. In C. bursa-pastoris accessions from western Eurasia, where the 2 species occur in partial sympatry, we find higher levels of nucleotide diversity than in accessions from eastern Eurasia, where C. rubella does not grow. Furthermore, haplotypes are shared across ploidy levels at 4 loci in western but not in eastern Eurasia. We test whether haplotype sharing is due to retention of ancestral polymorphism or due to hybridization and introgression using a coalescent-based isolation-with-migration model. In western but not in eastern Eurasia, there is evidence for unidirectional gene flow from C. rubella to C. bursa-pastoris. An independent estimate of the timing of dispersal of C. bursa-pastoris to eastern Eurasia indicates that it probably predated introgression. Our results show that polyploid speciation need not result in immediate and complete reproductive isolation, that postpolyploidization hybridization and introgression can contribute significantly to genetic variation in a newly formed polyploid, and that divergence population genetic analysis constitutes a powerful way of testing hypotheses on polyploid speciation. PMID- 18417486 TI - Molecular evolution of a primate-specific microRNA family. AB - Lineage-specific microRNA (miRNA) families may contribute to developmental novelties during evolution. However, little is known about the origin and evolution of new miRNA families. We report evidence of an Alu-mediated rapid expansion of miRNA genes in a previously identified primate-specific miRNA family, drawn from sequencing and comparative analysis of 9 diverse primate species. Evolutionary analysis reveals similar divergence among miRNA copies whether they are within or between species, lineage-specific gain and loss of miRNAs, and gene pseudolization in multiple species. These observations support a birth-and-death process of miRNA genes in this family, implicating functional diversification during primate evolution. In addition, both secondary structure conservation and reduced single nucleotide polymorphisms density attest to functional constraint of this family in primates. Finally, we observed preferential expression of miRNAs in human placenta and fetal brain, suggesting a functional importance of this family for primate development. PMID- 18417487 TI - Trends in antimicrobial susceptibility of Escherichia coli isolates from urology services in The Netherlands (1998-2005). AB - OBJECTIVES: An increase in antibiotic resistance of Escherichia coli, the most common pathogen in urinary tract infections (UTIs), is encountered worldwide. Optimal treatment of UTIs will contribute substantially to limit antibiotic use and antimicrobial resistance. This study determined trends in antimicrobial resistance of uropathogenic E. coli, which can be of use to optimize UTI guidelines. METHODS: During 1998-2005, E. coli from urine samples of patients attending urology services were collected in three regions in The Netherlands: north-east (NE, n = 1084), west (W, n = 1064) and south (S, n = 1212). The antibiotic susceptibility was determined using microbroth dilution following CLSI guidelines. E. coli ATCC 35218 and ATCC 25922 were used as reference strains. RESULTS: Amoxicillin resistance remained stable over time (37% to 47%), but was higher in the south (44%) compared with the other regions (40%; P < 0.02). Resistance to piperacillin increased from 4% (1998) to 32% (2005; P < 0.001), and resistance to fluoroquinolones increased from 6% to 13% (P < 0.01). Interregional differences were observed for resistance to piperacillin (NE 10%, W 12%, S 14%; P < 0.05) and to fluoroquinolones (NE 7%, W 13%, S 8%; P < 0.001). Trimethoprim +/- sulfamethoxazole resistance remained stable (27% to 37%), as did that of nitrofurantoin (4% to 9%). The percentage of strains with multidrug resistance (resistance to three or more groups of antibiotics) for each region increased over time (P < 0.05). CONCLUSIONS: Antibiotic resistance was fairly constant over time for most agents tested, except for piperacillin and the fluoroquinolones. Regional differences were observed for several compounds. National and regional surveillance of antibiotic resistance is important to keep therapeutic guidelines up-to-date and adequate for the treatment of resistant microorganisms. PMID- 18417488 TI - Forward-time simulations of non-random mating populations using simuPOP. AB - Computer simulations play an important role in studies of non-random mating populations. Because of implementation difficulties, only very limited types of non-random mating schemes are provided in the currently available simulation programs. Starting with version 0.8.5, simuPOP provides a few mating schemes that can be used to simulate arbitrary non-random mating models. This article describes the concepts and methods behind these mating schemes and demonstrates their uses in a few examples, including partial self-mating, positive assortative mating, non-random outbreeding, and simulation of overlapping generations in age structured populations. AVAILABILITY: simuPOP is freely available at http://simupop.sourceforge.net, distributed under a GPL license. Cited examples are in the doc/cookbook directory of a simuPOP distribution. PMID- 18417489 TI - IDMap: facilitating the detection of potential leads with therapeutic targets. AB - Pharmaceutical industry has been striving to reduce the costs of drug development and increase productivity. Among the many different attempts, drug repositioning (retargeting existing drugs) comes into the spotlight because of its financial efficiency. We introduce IDMap which predicts novel relationships between targets and chemicals and thus is capable of repositioning the marketed drugs by using text mining and chemical structure information. Also capable of mapping commercial chemicals to possible drug targets and vice versa, IDMap creates convenient environments for identifying the potential lead and its targets, especially in the field of drug repositioning. AVAILABILITY: IDMap executable and its user manual including color images are freely available to non-commercial users at http://www.equispharm.com/idmap PMID- 18417490 TI - Analysis and improvement of cyclotron thallium target room shield. AB - Because of high neutron and gamma-ray intensities generated during bombardment of a thallium-203 target, a thallium target-room shield and different ways of improving it have been investigated. Leakage of neutron and gamma ray dose rates at various points behind the shield are calculated by simulating the transport of neutrons and photons using the Monte Carlo N Particle transport computer code. By considering target-room geometry, its associated shield and neutron and gamma ray source strengths and spectra, three designs for enhancing shield performance have been analysed: a shielding door at the maze entrance, covering maze walls with layers of some effective materials and adding a shadow-shield in the target room in front of the radiation source. Dose calculations were carried out separately for different materials and dimensions for all the shielding scenarios considered. The shadow-shield has been demonstrated to be one suitable for neutron and gamma dose equivalent reduction. A 7.5-cm thick polyethylene shadow shield reduces both dose equivalent rate at maze entrance door and leakage from the shield by a factor of 3. PMID- 18417491 TI - Radon in soil gas--investigation and data standardisation at radon reference sites, Czech Republic. AB - Radon risk mapping at building sites in the Czech Republic indicate an advisable level of protection of buildings against the penetration of radon from the geological basement. Various techniques and instruments are used for the measurement of radon in soil gas. A national radon chamber and three radon reference sites in Central Bohemia enable radon data standardisation. Tests of field radon measurements and the reported radon activity concentration in soil gas at the reference sites are based on comparisons with other organisations and with a database of reference site measurements. The radon reference sites have been used for various investigations of radon in soil gas during the period 2000 07. PMID- 18417492 TI - Radon: a special case in radiation protection. AB - The conversion conventions of ICRP 65 are based on equality of detriment, not on dosimetry. They are derived from epidemiological studies on miners by comparing the risk of having fatal lung cancer with the detriment associated with a unit of exposure in ICRP 60. Things have moved on since ICRP 65 and the new scientific evidence (numerator change, denominator change and also the dosimetric approach in ICRP 66) is pointing away from ICRP 65 in the direction of the long established UNSCEAR conversion factor of 9 (nSv h(-1))/(Bq m(-3)) radon progeny exposure, which is 50% higher than the ICRP 65 conversion convention for members of the public. Anyhow, smoking, by the almost multiplicative relationship with radon, determines to a considerable extent the lung cancer risk. Although there is a fairly general consensus among health physicists that radon exposure constitutes the largest and most variable contribution to the population exposure from natural sources, they are divided between themselves on the numerical value of the risk estimates and on the need and urgency to incite the population to take action. This relaxed attitude to radon exposure is reflected in the regulatory approach, which is very much in line with the perceived risk by the population. PMID- 18417493 TI - Association of birth outcomes and maternal, school, and neighborhood characteristics with subsequent numeracy achievement. AB - This study investigated the relation between birth characteristics and numeracy attainment at age 8 years. Using a multilevel approach, the authors analyzed all non-Aboriginal singletons born in Western Australia who attended government schools and participated in a Western Australia-wide numeracy test in grade 3 between 1999 and 2005. Appropriateness of intrauterine growth was expressed as the proportion of optimal growth parameters for gestational duration, infant sex, and maternal height and parity, which was derived from a total population of births without risk factors for growth restriction. After the authors controlled for sociodemographic factors, term birth and proportion of optimal head circumference at birth were associated with higher numeracy scores. Increasing proportion of optimal birth length and being firstborn were associated with relatively higher numeracy scores among children born to mothers residing in the most educationally deprived area. The relative advantage of being born first was also higher for children born to single mothers. In contrast, higher Apgar scores and greater proportion of optimal birth weight were associated with a lower relative advantage for children born to single mothers. In summary, term birth and increased growth in head circumference and length are key birth characteristics associated with higher numeracy scores, especially among disadvantaged children. PMID- 18417494 TI - Waist circumference and mortality. AB - The authors examined the association between waist circumference and mortality among 154,776 men and 90,757 women aged 51-72 years at baseline (1996-1997) in the NIH-AARP Diet and Health Study. Additionally, the combined effects of waist circumference and body mass index (BMI; weight (kg)/height (m)(2)) were examined. All-cause mortality was assessed over 9 years of follow-up (1996-2005). After adjustment for BMI and other covariates, a large waist circumference (fifth quintile vs. second) was associated with an approximately 25% increased mortality risk (men: hazard ratio (HR) = 1.22, 95% confidence interval (CI): 1.15, 1.29; women: HR = 1.28, 95% CI: 1.16, 1.41). The waist circumference-mortality association was found in persons with and without prevalent disease, in smokers and nonsmokers, and across different racial/ethnic groups (non-Hispanic Whites, non-Hispanic Blacks, Hispanics, and Asians). Compared with subjects with a combination of normal BMI (18.5-<25) and normal waist circumference, those in the normal-BMI group with a large waist circumference (men: > or =102 cm; women: > or =88 cm) had an approximately 20% higher mortality risk (men: HR = 1.23, 95% CI: 1.08, 1.39; women: HR = 1.22, 95% CI: 1.09, 1.36). The finding that persons with a normal BMI but a large waist circumference had a higher mortality risk in this study suggests that increased waist circumference should be considered a risk factor for mortality, in addition to BMI. PMID- 18417495 TI - A regression approach for estimating multiday adverse health effects of PM10 when daily PM10 data are unavailable. AB - The authors propose a regression-based approach for obtaining multiday estimates of the adverse health effects of ambient particulate matter less than 10 microm in diameter (PM(10)) when daily PM(10) time-series data are unavailable. This situation is common in the United States, because most US cities take PM(10) measurements every 6 days. Current evidence suggests that adverse effects of PM(10) are not concentrated on a single day but rather are spread out over multiple days, so the unavailability of daily PM(10) data presents a problem for the estimation of these effects. The proposed model estimates weights that are used to construct a linear combination of single-lag PM(10) effect estimates obtained from the available PM(10) data. It is shown that this new approach provides estimates of the effect of PM(10) on mortality that have less bias and mean squared error than currently available methods. Application of this method to the US cities contained in the National Morbidity, Mortality, and Air Pollution Study database produces an estimated national average effect of PM(10) on nonaccidental mortality in persons over age 65 years, corresponding to a 0.32% increase per 10-microg/m(3) increment in PM(10). The estimated effects for cardiorespiratory mortality and other mortality are 0.34% and 0.22%, respectively. PMID- 18417496 TI - Nuclear organization in human sperm: preliminary evidence for altered sex chromosome centromere position in infertile males. AB - BACKGROUND: Many genetic defects with a chromosomal basis affect male reproduction via a range of different mechanisms. Chromosome position is a well known marker of nuclear organization, and alterations in standard patterns can lead to disease phenotypes such as cancer, laminopathies and epilepsy. It has been demonstrated that normal mammalian sperm adopt a pattern with the centromeres aligning towards the nuclear centre. The purpose of this study was to test the hypothesis that altered chromosome position in the sperm head is associated with male infertility. METHODS: The average nuclear positions of fluorescence in-situ hybridization signals for three centromeric probes (for chromosomes X, Y and 18) were compared in normoozoospermic men and in men with compromised semen parameters. RESULTS: In controls, the centromeres of chromosomes X, Y and 18 all occupied a central nuclear location. In infertile men the sex chromosomes appeared more likely to be distributed in a pattern not distinguishable from a random model. CONCLUSIONS: Our findings cast doubt on the reliability of centromeric probes for aneuploidy screening. The analysis of chromosome position in sperm heads should be further investigated for the screening of infertile men. PMID- 18417497 TI - Success rates and cost of a live birth following fresh assisted reproduction treatment in women aged 45 years and older, Australia 2002-2004. AB - BACKGROUND: The aim of this study was to calculate assisted reproductive technology (ART) success rates for fresh autologous and donor cycles in women aged > or = 45 and the resultant cost per live birth. METHODS: We performed a retrospective population-based study of 2339 ART cycles conducted in Australia, 2002-2004 to women aged > or = 45 years. The cost-outcome study was performed on fresh autologous treatment cycles. RESULTS: There were 1101 fresh autologous cycles initiated in women aged > or = 45, with a pregnancy rate of 1.9 per 100 initiated cycles. There were 21 women who achieved a clinical pregnancy with 15 (71%) ending in early pregnancy loss and 6 in live singleton births. The live birth rate following fresh autologous initiated cycles was 0.5% [95% confidence interval (CI): 0.1-1.0%]. Fresh donor recipients had an higher live birth rate of 19.1% (95% CI: 15.1-23.2) (odds ratio 43.2; 95% CI: 18.6-100.3) compared with women having fresh autologous cycles. The average cost of a live birth following fresh autologous cycles was 753,107 euros. CONCLUSIONS: The success rate of fresh autologous treatment for women aged > or = 45 years was < 1%. The very high cost of a live birth reflects a treatment failure rate of > 99%. The ART profession should counsel patients of the reality of the technology before the patients consent to treatment. PMID- 18417498 TI - Patient survival for all cancers combined as indicator of cancer control in Europe. AB - BACKGROUND: EUROCARE found marked differences in cancer survival across European populations, provoking extensive discussion as to the cause. We investigated the influence of socioeconomic indicators on survival, making use of the indicator population-based age-standardized and cancer site-standardized relative survival for all cancers combined (all cancer survival). METHODS: Bivariate correlation and multivariate regression analyses investigated relations between 1995 socioeconomic variables and all cancer survival in EUROCARE-3 patients from 19 European countries diagnosed 1990-94 and followed to 1999. RESULTS: Gross domestic product (GDP) and total national expenditure on health (TNEH) correlated highly with all cancer survival. Wealthy northern and western European countries had high survival; eastern European countries had low all cancer survival. GDP, TNEH, and number of computed tomography scanners per million--proxy of technological investment in cancer care--explained most survival differences. Low all cancer survival in the UK and Denmark compared to countries of similar wealth was closely related to fewer computed tomography scanners. Low all cancer survival in Poland compared to countries of similar wealth was also related to low TNEH. CONCLUSIONS: All cancer survival appears a useful and important indicator for monitoring countries' performance in cancer control. The most direct way for poorer European countries to improve all cancer survival would be to get richer; for richer countries more investment in health technology is important. However the sharply increasing costs of cancer care may render this impossible suggesting the need to radically rethink cancer control strategies. PMID- 18417499 TI - Improving continuity of care in an acute medical unit: initial outcomes. AB - BACKGROUND: The importance of acute medical units and their associated medical cover is stressed in current practice but there is a paucity of existing research to establish their impact on acute patient care. AIM: To assess the impact of a new medical admission process and associated medical cover on patient length of stay (LOS), direct discharge rates (DDR) (for admissions <24 and 48 h), daily discharge and readmission rates (RR). DESIGN: We performed a retrospective analysis of 3163 medical patients admitted before and after a ward was reconfigured to function as an acute medical unit (AMU), with a new on-call rota: 'consultant of the day' changing to 'consultant of the weekend', with aligned junior medical cover. METHODS: All medical admissions were analysed over three 2 month periods: two periods prior to the new AMU process (October to November, 2005 and June to July, 2006), and one period after the changes (October to Nov, 2006) which were made in August 2006. RESULTS: Average LOS was reduced from 8.6 and 9.3 for the two previous periods (June to July, 2006 and October to November, 2005) to 7.8 days for October to November, 2006, (P = 0.028). DDR for patients with a LOS under 24 and 48 h increased from 21.3% and 31.2% to 28.5% and 39.5%, respectively for both 24 h (P < 0.005) and 48 h LOS (P = 0.038). No significant difference in RR were observed (within 7 days) over the same periods. For admissions <48 h, the percentage of patients discharged increased for the Consultant-led teams (P < 0.006) before and after the new process. A statistically insignificant trend in relation to DDR was observed towards increased discharges over the weekend. DISCUSSION: The change in AMU process has resulted in improved DDR and patient length of stay, with no adverse effects on RR. PMID- 18417500 TI - Duplicated coronary sinus with a connecting branch. AB - A 48-year-old woman with class III heart failure and left bundle branch block underwent an implantation for cardiac resynchronization therapy. Right anterior oblique (RAO) view coronary sinus (CS) venography suggested the antero- and postero-lateral branches appeared to arise from the same vessel of a duplicated CS, but the antero-lateral branch arising from a different vessel was visualized via a connecting branch by the contrast injected into the vessel with the postero lateral branch, and the distal parts of the two vessels were superimposed in the RAO view. This unusual anomaly may have the potential risk for complications such as perforations. PMID- 18417501 TI - Acute radiation dermatitis and pneumonitis in Japanese breast cancer patients with whole breast hypofractionated radiotherapy compared to conventional radiotherapy. AB - OBJECTIVE: To evaluate acute morbidity, radiation dermatitis and pneumonitis, of Japanese patients treated with whole breast hypofractionated radiotherapy (RT) after breast-conserving surgery (BCS), compared to conventional RT. METHODS: Japanese patients who received whole breast RT after BCS between October 2003 and September 2006 were retrospectively reviewed. Patients who had selected the conventional or hypofractionated schedule received whole breast irradiation of 50 Gy in 25 fractions plus boost or 40 Gy in 16 fractions plus boost. Radiation dermatitis and symptomatic pneumonitis were graded according to the Common Terminology Criteria for Adverse Events version 3.0. RESULTS: Of 443 consecutive patients, 377 (85%) received the conventional schedule and 66 (15%) received the hypofractionated schedule. Of patients treated with the conventional schedule, Grade 0, 1, 2 and 3 radiation dermatitis were observed in 16 (4%), 278 (74%), 77 (20%) and 6 (2%), respectively. Of patients treated with the hypofractionated schedule, Grade 0, 1, 2 and 3 dermatitis were observed in 11 (17%), 49 (74%), 5 (8%) and 1 (1%), respectively. Grade 2-3 dermatitis by the hypofractionated schedule (9%) was observed less frequently than that by the conventional schedule (22%) (chi-square test; P = 0.016). Moreover, of patients treated with the conventional schedule, 4 (1%) had Grade 2 radiation pneumonitis. No patient treated with the hypofractionated schedule had symptomatic pneumonitis. CONCLUSIONS: Radiation dermatitis and pneumonitis in Japanese patients treated with the hypofractionated schedule is acceptable. Especially, radiation dermatitis by the hypofractionated schedule is milder than that by the conventional schedule. PMID- 18417502 TI - Docetaxel plus prednisolone for the treatment of metastatic hormone-refractory prostate cancer: a multicenter Phase II trial in Japan. AB - BACKGROUND: Docetaxel-based chemotherapy has been shown to be effective and well tolerated by Western patients with metastatic hormone-refractory prostate cancer (HRPC). This study was undertaken to assess the feasibility of docetaxel in combination with prednisolone in Japanese patients with HRPC. METHODS: Patients aged 50-74 years with measurable metastatic HRPC were included in this non comparative Phase II study. Treatment consisted of docetaxel 70 mg/m(2) once every 3 weeks plus prednisolone 5 mg twice daily, for a maximum of 10 cycles. The primary endpoint was overall tumor response rate, assessed by Response Evaluation Criteria in Solid Tumors; secondary endpoints included prostate-specific antigen (PSA) response and toxicity. RESULTS: A total of 43 patients were evaluable for efficacy and toxicity. The response rate was 44.2% (90% CI, 31.2-57.8%), with partial responses in 19/43 patients. The median duration of response was 19.3 weeks. PSA responses were recorded in 44.4% of patients (95% CI, 27.9-61.9%). The most common non-hematological adverse events (of any grade) possibly related to treatment were alopecia (88.4%), anorexia (65.1%) and fatigue (53.5%). Grade 3/4 leukopenia and neutropenia occurred in 81.4 and 93.0% of patients, respectively; however, the grade 3/4 rates of febrile neutropenia (16.3%) and infection without fever (14.0%) were lower. CONCLUSION: The combination of docetaxel and prednisolone was feasible and active in Japanese patients with HRPC, with a manageable adverse-event profile similar to that observed in Western patients. PMID- 18417503 TI - Spinal cord mechanisms of pain. AB - The spinal cord is the first relay site in the transmission of nociceptive information from the periphery to the brain. Sensory signals are transmitted from the periphery by primary afferent fibres into the dorsal horn of the spinal cord, where these afferents synapse with intrinsic spinal dorsal horn neurones. Spinal projection neurones then convey this information to higher centres in the brain, where non-noxious and noxious signals can be perceived. During nociceptive transmission, the output of the spinal cord is dependent on various spinal mechanisms which can either increase or decrease the activity of dorsal horn neurones. Such mechanisms include local excitatory and inhibitory interneurones, N-methyl-D-aspartate receptor activation, and descending influences from the brainstem, which can be both inhibitory and excitatory in nature. After nerve injury or conditions of inflammation, shifts can occur in these excitatory and inhibitory mechanisms which modulate spinal excitability, often resulting in the heightened response of dorsal neurones to incoming afferent signals, and increased output to the brain, a phenomenon known as central sensitization. In this review, we consider the ways in which spinal cord activity may be altered in chronic pain states. In addition, we discuss the spinal mechanisms which are targeted by current analgesics used in the management of chronic pain. PMID- 18417504 TI - Novel thin bronchoscope with a 1.7-mm working channel for peripheral pulmonary lesions. AB - In the present study, the authors evaluated the diagnostic utility of a novel thin bronchoscope with a 1.7-mm working channel for peripheral pulmonary lesions. A total of 118 patients were included in this prospective study. Bronchoscopic examination was performed using a 5.9-mm standard bronchoscope. If no visible endobronchial lesion was found, transbronchial biopsies were performed with 1.5 mm biopsy forceps under fluoroscopic guidance and the bronchus were washed with 10-20 mL of saline solution, using a prototype 3.5-mm thin bronchoscope with a 1.7-mm working channel. Endobronchial lesion was visualised with the standard bronchoscope in 16 patients, and the other 102 patients underwent biopsies with the thin bronchoscope. The mean bronchus levels reached with the standard bronchoscope and the thin bronchoscope were 2.3 and 4.3 generations, respectively. Endobronchial abnormality was revealed with the thin bronchoscope in a further 14 patients. Diagnostic material was obtained in 50 of 68 (74%) patients with malignant disease and 18 of 30 (60%) patients with benign disease. Four patients did not return to follow-up. The diagnostic yield was 57%, even in lesions <20 mm. There were no major complications. In conclusion, bronchoscopy using a 3.5-mm thin bronchoscope with a 1.7-mm working channel is useful and safe for the diagnosis of peripheral pulmonary lesions. PMID- 18417505 TI - Higher pulmonary artery pressure in children than in adults upon fast ascent to high altitude. AB - The response of pulmonary artery pressure to high altitude has not been studied in children. It is also not known whether the individual response is hereditary. Therefore, the response of pulmonary artery pressure to high altitude was measured in pre-pubertal children in comparison to that in their biological fathers. Echocardiography was performed at 450 m and over 3 days at 3,450 m. Systolic pulmonary artery pressure was estimated from the pressure gradient of tricuspid regurgitation. The increase in pulmonary artery pressure in children was greater than that in adults at day 1 of high altitude (15.5+/-9.1 versus 7.9+/-6.4 mmHg), but returned to adult levels on day 2. The increase in pulmonary artery pressure from low to high altitude of each child correlated with that in the father. Pre-pubertal children transiently develop greater pulmonary hypertension than their fathers when exposed to high altitude. The individual response of pulmonary pressure to high altitude seems to be at least partly hereditary. PMID- 18417506 TI - Interleukin 13, CD14, pet and tobacco smoke influence atopy in three Dutch cohorts: the allergenic study. AB - Studying gene-environment interactions may elucidate the complex origins of atopic diseases but requires large study populations. Pooling data from several cohort studies may help but may also obscure findings. Gene-environment interactions in atopy development were studied and the benefits of pooling data were evaluated. Haplotype-tagging polymorphisms in the genes interleukin (IL)13 and CD14 were genotyped in 3,062 children from the following birth cohorts: the Prevention and Incidence of Asthma and Mite Allergy (PIAMA) study; the Prevention of Asthma in Children (PREVASC) study; and the Child, Parent, Health, Focus on Lifestyle and Predisposition (KOALA) study, and tested for association with total and specific immunoglobulin (Ig)E and interaction with tobacco smoke and pet exposure at ages 1, 2, 4 and 8 yrs by analysis of variance, Chi-squared tests and regression analyses. At all ages, in IL13, minor alleles of rs1295685 and rs20541 were significantly associated with elevated IgE levels in pooled analyses. In CD14, the rs2569190-TT and rs2569191-CC genotypes associated with lower IgE and decreased risk of sensitisation at 4 and 8 yrs in children exposed to pets, with an opposite effect in nonexposed children. Findings for IL13 and CD14 were comparable in separate cohorts. The present study indicates that atopy is importantly influenced by interleukin 13 at age 1-8 yrs and by CD14 in interaction with pet exposure at ages 4 and 8 yrs. Additionally, pooled data improved effect estimates and genetic effects could be detected in interaction with important environmental factors. PMID- 18417507 TI - Reference values for peak flow and FEV1 variation in healthy schoolchildren using home spirometry. AB - Current reference values for diurnal peak flow variation in healthy children (median 8.2%; 95th percentile 31%) are so high that considerable overlap exists with those of asthmatic children. These values have been obtained using written peak flow diaries, which are unreliable. The aim of the present study was to obtain reliable reference values for the variation in peak flow and forced expiratory volume in one second (FEV(1)) in healthy schoolchildren using home spirometry with electronic data storage. Healthy schoolchildren (n = 204; 100 males) aged 6-16 yrs measured their peak flow and FEV(1) twice daily for 2 weeks using an electronic home spirometer. The variation in peak flow and FEV(1) were calculated as a diurnal amplitude as a percentage of the day's mean. The mean peak flow variation was 6.2% (95th percentile 12.3%) and the mean FEV(1) variation was 5.7% (95th percentile 11.8%). Using home spirometry with electronic data storage, healthy schoolchildren show considerably less peak flow and forced expiratory volume in one second variation than previously reported on the basis of written peak flow diaries. Being the 95th percentiles of the distributions in healthy children, a peak flow variation of 12.3% and an forced expiratory volume in one second variation of 11.8% are suggested as cut-off values for disease when using home spirometry. PMID- 18417508 TI - Smoke exposure, airway symptoms and exhaled nitric oxide in infants: the Generation R study. AB - The effect of pre- and post-natal smoke exposure on exhaled nitric oxide fraction (F(eNO)) in infants was evaluated and the association between respiratory symptoms and F(eNO) in the first 2 months of life was investigated. The Generation R study is a population-based, prenatally recruited birth cohort. Exposures were assessed by means of questionnaires prospectively administered during pregnancy and after birth. Successful off-line F(eNO) measurements during tidal breathing were obtained in 187 infants (median age 6.9 weeks). The association between possible determinants and log F(eNO) was investigated with multiple linear regression analysis. Infants exposed pre- and post-natally to smoke showed lower F(eNO) than infants exposed only after birth (geometric mean difference (95% confidence interval) 1.5 (1.0-2.1) ppb) and never-exposed infants (1.4 (1.0-1.8) ppb). F(eNO) was reduced in infants with severe upper respiratory symptoms compared with infants with nonsevere symptoms (1.6 (1.0-2.4) ppb). Infants with symptoms of the lower respiratory tract had lower F(eNO) than asymptomatic infants (1.2 (1.0-1.50) ppb). In conclusion, the nature of the association between smoke exposure and exhaled nitric oxide fraction is dependent on timing and intensity of exposure. The occurrence and the severity of respiratory symptoms in the first 2 months of life are associated with lower exhaled nitric oxide fraction. PMID- 18417509 TI - Selection of housekeeping genes for real-time PCR in atopic human bronchial epithelial cells. AB - The stability of housekeeping genes (HKGs) is critical when performing real-time quantitative PCR. To date, the stability of common HKGs has not been systematically compared in human airway epithelial cells (AEC) in normal and atopic subjects. Expression levels of 12 HKGs were measured in AECs from a cohort of 30 healthy atopic nonasthmatic or atopic asthmatic children. Gene expression stability was determined using three different Visual Basic for Applications applets (geNorm, NormFinder and BestKeeper). All 12 HKGs were expressed in AECs. However, the hypoxanthine ribosyltransferase and TATA-binding protein genes were excluded from further analysis due to low expression levels. The cyclophilin A gene was ranked the most stable by all three methods. The expression levels of the beta-actin and glyceraldehyde-3-phosphate dehydrogenase genes were significantly different between the three groups of patients, with atopic asthmatics showing the highest expression levels for both genes. The results suggest that the cyclophilin A gene is the most suitable housekeeping gene analysed for expression studies utilising uncultured bronchial airway epithelial cells from healthy and asthmatic children, and highlight the importance of validating housekeeping genes for each experimental model. PMID- 18417510 TI - Health belief model predicts adherence to CPAP before experience with CPAP. AB - Adherence to continuous positive airway pressure (CPAP) therapy for obstructive sleep apnoea (OSA) is often poor. Biomedical indices explain little of the variance in CPAP use. The present study tested a health belief model of adherence in order to determine the contribution of psychological constructs as compared to biomedical indices in the prediction of CPAP adherence. Consecutive patients (n = 77) newly diagnosed with OSA and naive to CPAP treatment (had never tried CPAP before) completed questionnaires at baseline (prior to CPAP treatment). The questionnaires assessed: outcome expectancy with treatment, self-efficacy, functional outcomes of sleepiness, and perceived risk of negative health outcomes. Physiological data were obtained from a standard clinical diagnostic sleep study. CPAP adherence was assessed at 4-month follow-up. Health belief model constructs alone explained 21.8% of the variance in CPAP adherence, whereas health belief model constructs and biomedical indices together explained 31.8% of the variance in CPAP adherence. The greatest proportion of CPAP adherence was explained by higher outcome expectancies with treatment, greater functional limitations as a result of sleepiness and lower risk perception. The results suggest that patients have developed beliefs and expectations about obstructive sleep apnoea and continuous positive airway pressure even before they try continuous positive airway pressure treatment. These beliefs and expectations predict the patients' adherence to effective therapy. PMID- 18417511 TI - Modulation of ozone-induced airway hyperresponsiveness and inflammation by interleukin-13. AB - The present study aimed to determine whether the T-helper cell type 2-derived cytokines, interleukin (IL)-4 and -13, can modulate the lung response to ozone exposure. IL-13(-/-), IL-4/13(-/-) and IL-13-overexpressing transgenic (Tg) mice were exposed to ozone (3 ppm; 3 h) or air. Wild-type (Wt) Balb/c mice and transgenic-negative littermates (IL-13Wt) were used as controls for gene deficient and IL-13Tg mice, respectively. IL-4/13(-/-) and IL-13(-/-) mice developed a lesser degree of ozone-induced airway hyperresponsiveness (AHR) while IL-13Tg mice developed a greater degree of AHR compared with ozone-exposed wild type or IL-13Wt mice, respectively. Ozone caused a time-dependent increase of bronchoalveolar lavage (BAL) neutrophils and macrophages in wild-type mice, maximal at 20-24 h, which was attenuated in the IL-13(-/-) and IL-4/13(-/-) mice. In IL-13Tg mice, there was a greater increase in BAL neutrophils after ozone exposure compared with IL-13Wt mice. Using quantitative real-time PCR, ozone induced mRNA expression for IL-6 and keratinocyte chemokine was further enhanced in IL-13(-/-) and IL-4/13(-/-) mice, and was inhibited in IL-13Tg mice. Macrophage inflammatory protein (MIP)-3alpha/CCL20 expression was enhanced after ozone exposure in wild-type mice, inhibited in IL-13(-/-) and IL-4/13(-/-) mice, while in IL-13Tg mice it was enhanced. A similar pattern of expression was observed with lipopolysaccharide-induced cytokine (LIX/CXCL5/ENA-78) expression. In conclusion, interleukin-13 augments ozone-induced airway hyperresponsiveness and neutrophilic inflammation, possibly through modulation of certain cytokines induced by ozone exposure. PMID- 18417512 TI - Impact of acute exposure to tobacco smoke on gelatinases in the bronchoalveolar space. AB - Clinical studies have indicated increased gelatinase activity in the airways of patients suffering from chronic obstructive pulmonary disease caused by tobacco smoke. The present study aimed to determine whether acute exposure to tobacco smoke per se causes a substantial and lasting impact on gelatinases and their inhibitors in the peripheral airways of atopic and nonatopic human subjects. Bronchoscopy with bronchoalveolar lavage (BAL) was performed on occasional smokers with and without atopy before and after smoking 10 cigarettes over a 48-h period. Samples from a group of never-smokers not exposed to tobacco smoke served as controls. Gelatinase identity and activity were measured using zymography, and gelatinase activity assay and concentrations of matrix metalloproteinase (MMP)-2, MMP-9, tissue inhibitor of MMP (TIMP)-1 and TIMP-2 were measured using ELISA. The results revealed no pronounced changes in identity, net activity or concentration of the gelatinases or changes in concentrations of TIMP-1 and TIMP-2 in BAL fluid before and after acute exposure to tobacco smoke. In conclusion, the present experimental study indicates that acute exposure to tobacco smoke does not cause any substantial impact on gelatinases or their inhibitors in the peripheral airways, irrespective of atopy status, a finding that is compatible with the fact that it takes many years of tobacco smoking to establish chronic obstructive pulmonary disease. PMID- 18417513 TI - The incidence of respiratory symptoms and sensitisation in baker apprentices. AB - The aim of the current study was to describe the incidence of respiratory symptoms and allergic sensitisation and the sequence of events leading to respiratory symptoms among Danish baker apprentices (BA). A total of 114 BA were surveyed over a 20-month period. Questionnaires were completed along with spirometric analysis and skin-prick tests to common and work-related allergens. Bronchial hyperresponsiveness (BHR) was determined at baseline and at the end of the follow-up period. The incidences of work-related rhinitis and asthma-like symptoms were 22.1 and 10.0 cases.100 person-yrs(-1), respectively. At 20 months the cumulative incidence proportion was 40.2 and 20.5% for rhinitis and asthma like symptoms, respectively. The cumulative incidence of occupational sensitisation was 6.1%. An increased risk of asthma-like symptoms was found in both atopic subjects and in females. In the BA with new onset respiratory symptoms, an increase in BHR from baseline was observed. Forced expiratory volume in one second and forced vital capacity did not change during the follow-up period. No relationship was observed between new sensitisation and new symptoms. In conclusion rhinitis- and asthma-like symptoms were found to develop commonly in Danish baker apprentices. The mechanism by which symptoms arose was perceived to reflect the development of an inflammation rather than the production of a specific immunoglobulin E pathway, as sensitisation to occupational allergens was rarely observed. Hence, respiratory symptoms and allergy may develop through separate pathways. PMID- 18417514 TI - NKG2D-dependent effector function of bronchial epithelium-activated alloreactive T-cells. AB - Allogeneic haematopoietic stem cell transplantation (SCT) has emerged as a curative therapeutic option. However, the role of graft-versus-host disease in lung injury after SCT has yet to be determined. In the present study, primary bronchial epithelial cells and the bronchial epithelial cell line BEAS-2B were used to investigate immune responses of allogeneic CD8+ T-cells directed against respiratory epithelial cells. Following stimulation with irradiated bronchial epithelial cells, CD8+ T-cells produced significant amounts of interferon-gamma, upregulated alloantigen activation markers and proliferated highly compared with T-cells stimulated with interleukin-2 alone. Furthermore, cytotoxicity assays demonstrated that bronchial epithelial cell-specific and granzyme B-mediated cytolytic activity was induced in CD8+ T-cells. Generation of natural killer (NK) T-cells, NK-like T-cells, cytokine-induced killer cells or lymphokine-activated killer cells could be excluded by phenotyping, culture conditions and neglectable lytic activity following stimulation with interleukin-2 alone. Inhibition experiments showed that lysis of bronchial epithelial cells was not major histocompatibility complex-I restricted, but depended on NK group 2 member D signalling; a stimulatory receptor initially shown to be expressed on NK cells. The present data imply that the respiratory epithelium has an antigen presenting function and directly alloactivates cytotoxic CD8+ T-cells that show nonclassical effector function. PMID- 18417515 TI - Increased urinary leukotriene E4 concentration in patients with eosinophilic pneumonia. AB - Although eosinophils produce cysteinyl leukotrienes (CysLTs) in large quantities, information on the relationship between CysLTs and eosinophilic pneumonia (EP) is lacking. Inflammatory mediator concentrations in urine were quantified to clarify the relationship between CysLT concentrations and EP severity. Leukotriene (LT)E(4), eosinophil-derived neurotoxin (EDN), 9alpha,11beta-prostaglandin F2 and LTB(4) glucuronide concentrations were quantified in the urine of: EP patients during acute exacerbation and clinical remission; asthmatic patients during acute exacerbation and under stable conditions; and healthy control subjects. The urinary LTE(4) and EDN concentrations of EP patients during acute exacerbation were significantly higher than those of asthmatic patients and healthy subjects, and decreased immediately during clinical remission. The urinary LTE(4) concentration was associated with the urinary EDN concentration of EP patients during acute exacerbation. The urinary LTE(4) concentration significantly correlated with the diffusing capacity of the lung for carbon monoxide in EP patients during acute exacerbation. The increased urinary concentrations of leukotriene and eosinophil-derived neurotoxin were associated with acute exacerbation in eosinophilic pneumonia patients. The increased leukotriene concentration significantly correlated with diffusing capacity of the lung for carbon monoxide, suggesting that the monitoring of leukotriene concentration may aid in the management of eosinophilic pneumonia patients. PMID- 18417516 TI - Changes in exercise haemodynamics during treatment in pulmonary arterial hypertension. AB - Improvement in exercise capacity may not be associated with significant changes in resting pulmonary haemodynamics. The present prospective study documented the relationships between improvement in 6-min walking distance (6MWD) and changes in resting and exercise pulmonary haemodynamics after treatment in patients with idiopathic pulmonary arterial hypertension (IPAH). A total of 42 IPAH patients underwent supine submaximal exercise haemodynamic assessments at baseline and after 5+/-2 months of therapy. The 6MWD, the corresponding chronotropic response (peak minus resting heart rate), and resting and exercise haemodynamic variables were analysed. The isoflow was defined as the lowest of the pre- and post treatment peak cardiac index (CI). The extrapolated isoflow mean pulmonary artery pressure ((pa)) was used to characterise changes in (pa)-CI regression lines following treatment. Patients were given bosentan (n = 28), epoprostenol (n = 12) or both. The 6MWD increased significantly, from 399+/-88 to 442+/-86 m. On univariate analysis, changes in 6MWD correlated with changes in isoflow (pa), chronotropic response, resting haemodynamics (CI, pulmonary vascular resistance and mixed venous oxygen saturation) and exercise haemodynamics (peak CI). On multivariate analysis, only changes in isoflow (pa) and chronotropic response were independently associated with changes in 6MWD. Improvement in exercise tolerance with chronic therapy is independently related to improvement in pulmonary haemodynamics measured in exercise but not in resting conditions. PMID- 18417517 TI - Prospective evaluation of risk of vertebral fractures using quantitative ultrasound measurements and bone mineral density in a population-based sample of postmenopausal women: results of the Basel Osteoporosis Study. AB - OBJECTIVE: Prospective studies have shown that quantitative ultrasound (QUS) techniques predict the risk of fracture of the proximal femur with similar standardised risk ratios to dual-energy x-ray absorptiometry (DXA). Few studies have investigated these devices for the prediction of vertebral fractures. The Basel Osteoporosis Study (BOS) is a population-based prospective study to assess the performance of QUS devices and DXA in predicting incident vertebral fractures. METHODS: 432 women aged 60-80 years were followed-up for 3 years. Incident vertebral fractures were assessed radiologically. Bone measurements using DXA (spine and hip) and QUS measurements (calcaneus and proximal phalanges) were performed. Measurements were assessed for their value in predicting incident vertebral fractures using logistic regression. RESULTS: QUS measurements at the calcaneus and DXA measurements discriminated between women with and without incident vertebral fracture, (20% height reduction). The relative risks (RRs) for vertebral fracture, adjusted for age, were 2.3 for the Stiffness Index (SI) and 2.8 for the Quantitative Ultrasound Index (QUI) at the calcaneus and 2.0 for bone mineral density at the lumbar spine. The predictive value (AUC (95% CI)) of QUS measurements at the calcaneus remained highly significant (0.70 for SI, 0.72 for the QUI, and 0.67 for DXA at the lumbar spine) even after adjustment for other confounding variables. CONCLUSIONS: QUS of the calcaneus and bone mineral density measurements were shown to be significant predictors of incident vertebral fracture. The RRs for QUS measurements at the calcaneus are of similar magnitude as for DXA measurements. PMID- 18417518 TI - Total endovascular aortic arch reconstruction via fenestration in situ with cerebral circulatory support: an acute experimental study. AB - The aim of this experimental study is to evaluate the feasibility of endovascular repair of the complete aortic arch by using novel fenestration devices with simultaneous support of the cerebral circulation. Two fresh human cadavers and five Yorkshire pigs were used for the experiments. In human cadavers the thoracic aorta was pressurized using a roller pump to simulate the circulation. In animal experiments right femoral artery to right distal carotid artery bypass circuit was achieved in order to support the cerebral circulation during the stent graft deployment, fenestration and conduit fixation procedures. Commercially available Valiant Thoracic Stent Grafts, covered stents, steerable guiding catheters and dilatation balloons were used. Stent grafts were deployed successfully and two fenestrations and one conduit implantation were achieved in each cadaver. All animals survived the stent graft implantation, fenestration and conduit implantation procedures. Cadaver dissection and necropsy of the animals revealed good fixation of the conduits into the fenestrated segments of the stent graft. Endovascular repair of the total aortic arch via in situ fenestration of the stent graft using cerebral circulatory support seems to be feasible and safe. Further studies are required before clinical adoption of this procedure. PMID- 18417519 TI - Short-term transcutaneous electrical nerve stimulation after cardiac surgery: effect on pain, pulmonary function and electrical muscle activity. AB - This study aimed to evaluate the effectiveness of transcutaneous electrical nerve stimulation (TENS) for treatment of postoperative pain in patients who underwent cardiac surgery. In addition, we sought to determine whether TENS would be related to improved pulmonary function and muscle electrical activity in this patient population. Forty-five patients, 32 males and 13 females, aged 41-74 years were randomly allocated to receive TENS (n=23) or sham treatment (n=22) during 4 h on the third postoperative day. A 0-10 visual analogic scale was used to assess pain; lung function was evaluated by spirometry and surface electromyography (n=10 in each group) was used to quantify electrically-induced muscle activity (trapezius and pectoralis major). TENS was associated with significant reductions on spontaneous and cough-induced postoperative pain as compared to sham (P<0.05). There was also improvement in chest wall-pulmonary mechanics after TENS with proportional increases in tidal volume and vital capacity (P<0.05). In addition, electrical activity of both muscle groups was enhanced after TENS, but not post sham (P<0.05). TENS is a valuable strategy to alleviate postoperative pain following cardiac surgery with positive effects on pulmonary ventilatory function and electrical activity of thoracic and girdle muscles. PMID- 18417520 TI - Screening for MRSA. PMID- 18417523 TI - Health department's denial of free health care to asylum seekers was unlawful, judge says. PMID- 18417521 TI - Impact of rapid screening tests on acquisition of meticillin resistant Staphylococcus aureus: cluster randomised crossover trial. AB - OBJECTIVE: To determine whether introducing a rapid test for meticillin resistant Staphylococcus aureus (MRSA) screening leads to a reduction in MRSA acquisition on hospital general wards. DESIGN: Cluster randomised crossover trial. SETTING: Medical, surgical, elderly care, and oncology wards of a London teaching hospital on two sites. MAIN OUTCOME MEASURE: MRSA acquisition rate (proportion of patients negative for MRSA who became MRSA positive). PARTICIPANTS: All patients admitted to the study wards who were MRSA negative on admission and screened for MRSA on discharge. INTERVENTION: Rapid polymerase chain reaction based screening test for MRSA compared with conventional culture. RESULTS: Of 9608 patients admitted to study wards, 8374 met entry criteria and 6888 had full data (82.3%); 3335 in the control arm and 3553 in the rapid test arm. The overall MRSA carriage rate on admission was 6.7%. Rapid tests led to a reduction in median reporting time from admission, from 46 to 22 hours (P<0.001). Rapid testing also reduced the number of inappropriate pre-emptive isolation days between the control and intervention arms (399 v 277, P<0.001). This was not seen in other measurements of resource use. MRSA was acquired by 108 (3.2%) patients in the control arm and 99 (2.8%) in the intervention arm. When predefined confounding factors were taken into account the adjusted odds ratio was 0.91 (95% confidence interval 0.61 to 1.234). Rates of MRSA transmission, wound infection, and bacteraemia were not statistically different between the two arms. CONCLUSION: A rapid test for MRSA led to the quick receipt of results and had an impact on bed usage. No evidence was found of a significant reduction in MRSA acquisition and on these data it is unlikely that the increased costs of rapid tests can be justified compared with alternative control measures against MRSA. TRIAL REGISTRATION: Clinical controlled trials ISRCTN75590122 [controlled-trials.com]. PMID- 18417524 TI - Efficacy and safety of rituximab in a patient with active rheumatoid arthritis and chronic disseminated pulmonary aspergillosis and history of tuberculosis. PMID- 18417525 TI - Infliximab effective in steroid-dependent juvenile eosinophilic fasciitis. PMID- 18417526 TI - Associations of DNase IV polymorphisms with autoantibodies in patients with systemic lupus erythematosus. AB - OBJECTIVES: The aim of this study was to investigate genetic polymorphisms of DNase IV and their relationship with SLE and various autoantibodies present in SLE patients. METHODS: A total of 532 SLE patients and 521 healthy controls belonging to the Korean population were enrolled into this study. Sequencing of the entire coding region of the DNase IV gene (including the promoter region) was carried out using a DNA analyser. Autoantibodies including anti-Sm, anti-Ro, anti La, anti-RNP and anti-dsDNA were determined either by indirect immunofluorescence or double immunodiffusion methods. Multiple logistic regression analysis was performed to examine the genetic association with SLE and autoantibodies. RESULTS: We found three single-nucleotide polymorphisms (SNPs): -2753G-->A, +147T ->G (Gly49Gly) and +1466G-->T. The -2753G-->A and +147T-->G (Gly49Gly) SNPs were selected for larger scale genotyping based on linkage disequilibria and haplotype tagging status. Although the -2753G-->A SNP was more common than the +147T-->G (Gly49Gly) SNP (frequencies: 0.330 and 0.002, respectively), its association with the risk of SLE was not statistically significant. However, -2753G-->A SNP was significantly associated with the production of anti-Sm antibody [odds ratio (95% CI): co-dominant model, 1.89 (1.28-2.79); dominant model, 2.17 (1.20-3.90) and recessive model, 2.62 (1.33-5.17)]. CONCLUSIONS: We did not find significant relationships between DNase IV polymorphisms and the risk of SLE, but the association of the common -2753G-->A allele in the promoter region with the production of anti-Sm antibody implicates DNase IV as a putative candidate gene of SLE. PMID- 18417527 TI - Association between duration of symptoms and severity of disease at first presentation to paediatric rheumatology: results from the Childhood Arthritis Prospective Study. AB - OBJECTIVES: To study the association between disease severity at first presentation to paediatric rheumatology (PRh) and length of time since symptom onset in children recruited to the Childhood Arthritis Prospective Study. METHODS: Children or=2 weeks were recruited from five UK hospitals. Data including demographics, disease features, Childhood Health Assessment Questionnaire (CHAQ), physician and parent global assessment and blood tests were collected at the first appointment with PRh (baseline). The association between symptom duration (defined as time from first reported symptom onset to presentation at PRh) and baseline disease characteristics was evaluated using non-parametric descriptive statistics and multivariable logistic regression analyses. RESULTS: Five hundred and seven children (65% female) were included: median age at onset was 6.8 yrs. Two hundred and thirty-three had oligoarthritis, 68 had RF-negative polyarthritis, 27 had systemic onset arthritis and 29 had arthritis that was not JIA. The median symptom duration was 4.6 months. Median symptom duration was shortest for children presenting with systemic arthritis (1.6 months) and longest for those with PsA (8.6 months). Children with a longer duration of symptoms were older and had higher median active joint counts but lower median ESR. Symptom duration did not correlate with CHAQ score at presentation. CONCLUSIONS: Children who have systemic arthritis had the shortest delay to PRh presumably because they are profoundly unwell. Children with joint pain/stiffness but normal ESR had longer delays suggesting that if blood tests do not indicate inflammation, the diagnosis of JIA may be overlooked. PMID- 18417528 TI - Costing juvenile idiopathic arthritis: examining patient-based costs during the first year after diagnosis. AB - OBJECTIVES: There are few data on the treatment patterns and associated cost of treating children with inflammatory arthritis including juvenile idiopathic arthritis (JIA), in the short or long term. The aim of this study was to obtain patient-based costs for treating children with JIA in the UK, in the first year from diagnosis and from the secondary health care payer perspective. METHODS: The Childhood Arthritis Prospective Study (CAPS) is an ongoing longitudinal study recruiting children with inflammatory arthritis from four UK hospital centres. Included children are newly diagnosed, 12 years of employment (chi(2) for trend = 5.45; p<0.05). A significant (p = 0.04) trend was confirmed by Poisson regression. No reduced risks were found for other forms of cancer, nor for those working with other tasks. CONCLUSIONS: The results support previous reports that a high and prolonged exposure to cotton dust and other endotoxin-containing organic dusts is related to a lower risk of lung cancer. There was no indication of a reduced risk for other forms of cancer. PMID- 18417555 TI - The impact of components of fine particulate matter on cardiovascular mortality in susceptible subpopulations. AB - BACKGROUND: Several studies have demonstrated associations between daily mortality and ambient particulate matter less than 2.5 microns in diameter (fine particles or PM2.5). Few, however, have examined the relative toxicities of PM2.5 constituents, including elemental carbon and organic carbon (EC and OC, respectively), nitrates and transition metals. There is also little information about whether associations between PM2.5 constituents and mortality are modified by socioeconomic and demographic factors. AIM: To examine associations of daily cardiovascular mortality with PM2.5 and its constituents after stratification by gender, race/ethnicity and education, using data from six California counties during 2000 to 2003. METHODS: The association of daily counts of cardiovascular mortality with PM2.5 components was analysed using time-series regression analyses. Poisson models with natural splines were used to control for time varying covariates such as season and weather. Separate models were run after stratification by gender, race/ethnicity (White, Hispanic, Black) and education (high school graduation or not). Models were run for each county and results were combined using random effects meta-analysis. RESULTS: Daily counts of cardiovascular mortality were associated with PM2.5 and several of its species including EC, OC, nitrates, sulphates, potassium, copper and iron. For many of these species, there were significantly higher effect estimates among those with lower educational attainment and Hispanic individuals. For example, while essentially no association was observed for individuals who graduated from high school, an interquartile change in several of the components of PM2.5 was associated with a 3-5% increase in daily mortality among non-high school graduates. CONCLUSION: There is evidence that several PM2.5 constituents may represent important contributors to cardiovascular mortality. Many of these constituents are generated by motor vehicles, especially those with diesel engines, and by residential wood combustion. In addition, factors associated with low educational attainment may increase susceptibility to PM2.5 and its components. PMID- 18417557 TI - Psychosocial factors at work and risk of depression: a systematic review of the epidemiological evidence. AB - OBJECTIVES: Major depression is a leading cause of psychiatric morbidity and may be influenced by psychosocial factors in the workplace, although evidence so far remains circumstantial. This paper reviews follow-up studies addressing the risk of major depression and depressive symptoms relative to psychosocial stressors in the working environment and evaluates the evidence for causality. METHOD: Follow up studies were identified by a systematic Medline search combining search terms for the outcome and measures of job-related psychosocial factors. The quality of the studies was evaluated using 22 criteria related to their potential for bias and confounding. RESULTS: Sixteen company or population-based studies including some 63,000 employees were identified. Validated multi-item scales were used to measure perceived psychosocial stressors in most of the studies. Major depression was defined by clinical criteria in seven studies and by symptom scales in another seven. The follow-up period ranged from 1 to 13 years. The prevalence of depressive disorder varied substantially, suggesting a high degree of study heterogeneity. The adjusted relative risk for onset of a major depressive episode according to job stressors ranged from 0.5 to 1.5 in 44 of 61 reported associations with various psychosocial factor dimensions. Associations were strongest and most consistent for job strain defined as high demand and low decision latitude among men. Most studies shared common limitations such as lack of independent measures of exposure and outcome and potential confounding. Although a meta-analysis would technically be possible, heterogeneity across studies evidenced by variation in the prevalence of depression made this unfeasible. CONCLUSION: This review provides consistent findings that perception of adverse psychosocial factors in the workplace is related to an elevated risk of subsequent depressive symptoms or major depressive episode; however, methodological limitations preclude causal inference. Studies implementing objective measures of job stressors or independent outcome ascertainment are warranted. PMID- 18417556 TI - Meta-analysis of benzene exposure and non-Hodgkin lymphoma: biases could mask an important association. AB - OBJECTIVES: Benzene is a widely recognised cause of leukaemia but its association with non-Hodgkin's lymphoma (NHL) is less well established. The goal of this project is to review the current published literature on this association. METHODS: We performed a meta-analysis of cohort and case-control studies of benzene exposure and NHL and a meta-analysis of NHL and refinery work, a potential source of benzene exposure. RESULTS: In 22 studies of benzene exposure, the summary relative risk for NHL was 1.22 (95% CI 1.02 to 1.47; one-sided p value = 0.01). When studies that likely included unexposed subjects in the "exposed" group were excluded, the summary relative risk increased to 1.49 (95% CI 1.12 to 1.97, n = 13), and when studies based solely on self-reported work history were excluded, the relative risk rose to 2.12 (95% CI 1.11 to 4.02, n = 6). In refinery workers, the summary relative risk for NHL in all 21 studies was 1.21 (95% CI 1.00 to 1.46; p = 0.02). When adjusted for the healthy worker effect, this relative risk estimate increased to 1.42 (95% CI 1.19 to 1.69). CONCLUSIONS: The finding of elevated relative risks in studies of both benzene exposure and refinery work provides further evidence that benzene exposure causes NHL. In addition, the finding of increased relative risks after removing studies that included unexposed or lesser exposed workers in "exposed" cohorts, and increased relative risk estimates after adjusting for the healthy worker effect, suggest that effects of benzene on NHL might be missed in occupational studies if these biases are not accounted for. PMID- 18417558 TI - Lung cancer and exposure to arsenic in rural Bangladesh. AB - BACKGROUND: Arsenic is a known carcinogen but the risk of lung cancer from the widespread contamination of drinking water in rural Bangladesh has not been estimated. OBJECTIVES: To determine whether estimated exposure of villagers in Bangladesh to arsenic in drinking water differed between those with lung cancer and those with non-malignant lesions. METHODS: Data were obtained from 7286 subjects who underwent lung biopsy in 2003-2006 at a diagnostic centre taking referrals from throughout Bangladesh. Analysis was limited to 5372 people living in villages for the last 10 years who reported using tube well water. Of these, 3223 with a primary lung tumour were enrolled as cases and 1588 with non malignant lesions as referents in an unmatched analysis. Arsenic exposure was estimated by average concentrations for each of 64 districts. Logistic regression was used to test the effects of age, arsenic and smoking on risk and to investigate relationship to cell type. RESULTS: Male cases were older than referents and more likely to smoke, to smoke >20 units/day and to smoke bidi small, hand-rolled cigarettes. Odds ratios for lung cancer increased steadily with mean arsenic concentration, but the confidence interval excluded 1.0 only at concentrations >100 mug/l (OR 1.45, 95% CI 1.16 to 1.80). This trend was seen only in smokers where the increased risk at >100 mug/l was 1.65 (95% CI 1.25 to 2.18). A similar trend was seen in women smokers. Squamous cell lung cancer was more frequent in smokers and, having adjusted for smoking, in districts with arsenic concentrations >100 mug/l. CONCLUSIONS: Among Bangladeshis who smoke, those whose drinking water is contaminated with arsenic at concentrations >100 mug/l are at increased risk of lung cancer. With high levels of exposure misclassification and short latency of exposure, the study cannot estimate or exclude the likely long term risk in non-smokers and at lower arsenic concentrations. PMID- 18417559 TI - Chronic health problems and risk of accidental injury in the workplace: a systematic literature review. AB - OBJECTIVES: To investigate whether common important health conditions and their treatments increase risks of occupational injury. METHODS: A systematic search was conducted of MEDLINE, EMBASE and PsycINFO databases from inception to November 2006 employing terms for occupational injury, medications, and a broad range of diseases and impairments. Papers related solely to driving, alcohol, or substance abuse were excluded, as were studies that did not allow analysis of injury risk. For each paper that was retrieved we abstracted standard information on the population, design, exposure(s), outcome(s), response rates, confounders and effect estimates; and rated the quality of information provided. RESULTS: We found 38 relevant papers (33 study populations): 16 studies were of cross sectional design, 13 were case-control and 4 were prospective. The overall quality was rated as excellent for only two studies. Most commonly investigated were problems of hearing (15 studies), mental health (11 studies) and vision (10 studies). For impaired hearing, neurotic illness, diabetes, epilepsy and use of sedating medication there were moderate positive associations with occupational injury (odds ratios 1.5-2.0), but there were major gaps in the evidence base. Studies covering vision did not present risks by category of eye disease; no evidence was found on psychotic illness; for diabetes, epilepsy and cardiovascular disease there were remarkably few papers; studies seldom distinguished risks by sub-category of external cause or anatomical site and nature of injury; and exposures and outcomes were mostly ascertained by self report at a single time point, with a lack of clarity about exposure timings. CONCLUSION: Improved research is needed to define the risks of occupational injury arising from common health complaints and treatments. Such research should delineate exposures and outcomes in more detail, and ensure by design that the former precede the latter. PMID- 18417560 TI - A randomised controlled trial on whether a participatory ergonomics intervention could prevent musculoskeletal disorders. AB - OBJECTIVES: To examine the efficacy of a participatory ergonomics intervention in preventing musculoskeletal disorders among kitchen workers. Participatory ergonomics is commonly recommended to reduce musculoskeletal disorders, but evidence for its effectiveness is sparse. METHODS: A cluster randomised controlled trial among the 504 workers of 119 kitchens in Finland was conducted during 2002-2005. Kitchens were randomised to an intervention (n = 59) and control (n = 60) group. The duration of the intervention that guided the workers to identify strenuous work tasks and to seek solutions for decreasing physical and mental workload, was 11 to 14 months. In total, 402 ergonomic changes were implemented. The main outcome measures were the occurrence of and trouble caused by musculoskeletal pain in seven anatomical sites, local fatigue after work, and sick leave due to musculoskeletal disorders. Individual level data were collected by a questionnaire at baseline and every 3 months during the intervention and 1 year follow-up period. All response rates exceeded 92%. RESULTS: No systematic differences in any outcome variable were found between the intervention and control groups during the intervention or during the 1-year follow-up. CONCLUSIONS: The intervention did not reduce perceived physical work load and no evidence was found for the efficacy of the intervention in preventing musculoskeletal disorders among kitchen workers. It may be that a more comprehensive redesign of work organisation and processes is needed, taking more account of workers' physical and mental resources. PMID- 18417561 TI - Kaposi's sarcoma-associated herpesvirus-encoded LANA can interact with the nuclear mitotic apparatus protein to regulate genome maintenance and segregation. AB - Kaposi's sarcoma-associated herpesvirus (KSHV) genomes are tethered to the host chromosomes and partitioned faithfully into daughter cells with the host chromosomes. The latency-associated nuclear antigen (LANA) is important for segregation of the newly synthesized viral genomes to the daughter nuclei. Here, we report that the nuclear mitotic apparatus protein (NuMA) and LANA can associate in KSHV-infected cells. In synchronized cells, NuMA and LANA are colocalized in interphase cells and separate during mitosis at the beginning of prophase, reassociating again at the end of telophase and cytokinesis. Silencing of NuMA expression by small interfering RNA and expression of LGN and a dominant negative of dynactin (P150-CC1), which disrupts the association of NuMA with microtubules, resulted in the loss of KSHV terminal-repeat plasmids containing the major latent origin. Thus, NuMA is required for persistence of the KSHV episomes in daughter cells. This interaction between NuMA and LANA is critical for segregation and maintenance of the KSHV episomes through a temporally controlled mechanism of binding and release during specific phases of mitosis. PMID- 18417562 TI - Rotavirus infection accelerates type 1 diabetes in mice with established insulitis. AB - Infection modulates type 1 diabetes, a common autoimmune disease characterized by the destruction of insulin-producing islet beta cells in the pancreas. Childhood rotavirus infections have been associated with exacerbations in islet autoimmunity. Nonobese diabetic (NOD) mice develop lymphocytic islet infiltration (insulitis) and then clinical diabetes, whereas NOD8.3 TCR mice, transgenic for a T-cell receptor (TCR) specific for an important islet autoantigen, show more rapid diabetes onset. Oral infection of infant NOD mice with the monkey rotavirus strain RRV delays diabetes development. Here, the effect of RRV infection on diabetes development once insulitis is established was determined. NOD and NOD8.3 TCR mice were inoculated with RRV aged > or = 12 and 5 weeks, respectively. Diabetes onset was significantly accelerated in both models (P < 0.024), although RRV infection was asymptomatic and confined to the intestine. The degree of diabetes acceleration was related to the serum antibody titer to RRV. RRV infected NOD mice showed a possible trend toward increased insulitis development. Infected males showed increased CD8(+) T-cell proportions in islets. Levels of beta-cell major histocompatibility complex class I expression and islet tumor necrosis factor alpha mRNA were elevated in at least one model. NOD mouse exposure to mouse rotavirus in a natural experiment also accelerated diabetes. Thus, rotavirus infection after beta-cell autoimmunity is established affects insulitis and exacerbates diabetes. A possible mechanism involves increased exposure of beta cells to immune recognition and activation of autoreactive T cells by proinflammatory cytokines. The timing of infection relative to mouse age and degree of insulitis determines whether diabetes onset is delayed, unaltered, or accelerated. PMID- 18417563 TI - Comparative efficacy of neutralizing antibodies elicited by recombinant hemagglutinin proteins from avian H5N1 influenza virus. AB - Although the human transmission of avian H5N1 virus remains low, the prevalence of this highly pathogenic infection in avian species underscores the need for a preventive vaccine that can be made without eggs. Here, we systematically analyze various forms of recombinant hemagglutinin (HA) protein for their potential efficacy as vaccines. Monomeric, trimeric, and oligomeric H5N1 HA proteins were expressed and purified from either insect or mammalian cells. The immunogenicity of different recombinant HA proteins was evaluated by measuring the neutralizing antibody response. Neutralizing antibodies to H5N1 HA were readily generated in mice immunized with the recombinant HA proteins, but they varied in potency depending on their multimeric nature and cell source. Among the HA proteins, a high-molecular-weight oligomer elicited the strongest antibody response, followed by the trimer; the monomer showed minimal efficacy. The coexpression of another viral surface protein, neuraminidase, did not affect the immunogenicity of the HA oligomer, as expected from the immunogenicity of trimers produced from insect cells. As anticipated, HA expressed in mammalian cells without NA retained the terminal sialic acid residues and failed to bind alpha2,3-linked sialic acid receptors. Taken together, these results suggest that recombinant HA proteins as individual or oligomeric trimers can elicit potent neutralizing antibody responses to avian H5N1 influenza viruses. PMID- 18417564 TI - Glycoproteins required for entry are not necessary for egress of pseudorabies virus. AB - In the current perception of the herpesvirus replication cycle, two fusion processes are thought to occur during entry and nuclear egress. For penetration, glycoproteins gB and gH/gL have been shown to be essential, whereas a possible role of these glycoproteins in nuclear egress remains unclear. Viral envelope glycoproteins have been detected by immunolabeling in the nuclear membrane as well as in primary enveloped particles in several herpesviruses, indicating that they might be involved in the fusion process. Moreover, a herpes simplex virus type 1 mutant simultaneously lacking gB and gH was described to be deficient in nuclear egress (A. Farnsworth, T. W. Wisner, M. Webb, R. Roller, G. Cohen, R. Eisenberg, and D. C. Johnson, Proc. Natl. Acad. Sci. USA 104:10187-10192, 2007). To analyze the situation in the related alphaherpesvirus pseudorabies virus (PrV), mutants carrying single and double deletions of glycoproteins gB, gD, gH, and gL were constructed and characterized. We show here that the simultaneous deletion of gB and gD, gB and gH, gD and gH, or gH and gL has no detectable effect on PrV egress, implying that none of these glycoproteins either singly or in the tested combinations is required for nuclear egress. In addition, immunolabeling studies using different mono- or polyclonal sera raised against various PrV glycoproteins did not reveal the presence of viral glycoproteins in the inner nuclear membrane or in primary virions. Thus, our data strongly suggest that different fusion mechanisms are active during virus entry and egress. PMID- 18417565 TI - RUNX1 permits E4orf6-directed nuclear localization of the adenovirus E1B-55K protein and associates with centers of viral DNA and RNA synthesis. AB - The localization of the adenovirus E1B-55K-E4orf6 protein complex is critical for its function. Prior studies demonstrated that E4orf6 directs the nuclear localization of E1B-55K in human cells and in rodent cells that contain part of human chromosome 21. We show here that the relevant activity on chromosome 21 maps to RUNX1. RUNX1 proteins are transcription factors that serve as scaffolds for the assembly of proteins that regulate transcription and RNA processing. After transfection, the RUNX1a, RUNX1b, and RUNX1-DeltaN variants allowed E4orf6 directed E1B-55K nuclear localization. The failure of RUNX1c to allow nuclear colocalization was relieved by the deletion of amino-terminal residues of this protein. In the adenovirus-infected mouse cell, RUNX1 proteins were localized to discrete structures about the periphery of viral replication centers. These sites are enriched in viral RNA and RNA-processing factors. RUNX1b and RUNX1a proteins displaced E4orf6 from these sites. The association of E1B-55K at viral replication centers was enhanced by the RUNX1a and RUNX1b proteins, but only in the absence of E4orf6. In the presence of E4orf6, E1B-55K occurred in a perinuclear cytoplasmic body resembling the aggresome and was excluded from the nucleus of the infected mouse cell. We interpret these findings to mean that a dynamic relationship exists between the E4orf6, E1B-55K, and RUNX1 proteins. In cooperation with E4orf6, RUNX1 proteins are able to modulate the localization of E1B-55K and even remodel virus-specific structures that form at late times of infection. Subsequent studies will need to determine a functional consequence of the interaction between E4orf6, E1B-55K, and RUNX1. PMID- 18417566 TI - Cell-type-specific tyrosine phosphorylation of the herpes simplex virus tegument protein VP11/12 encoded by gene UL46. AB - Cytotoxic T lymphocytes (CTL) and natural killer (NK) cells play key roles in limiting herpesvirus infections; consequently, many herpesviruses, including herpes simplex virus (HSV), have evolved diverse strategies to evade and/or disarm these killer lymphocytes. Previous studies have shown that CTL and NK cells are functionally inactivated following contact with HSV-infected fibroblasts. During studies of the mechanisms involved, we discovered that HSV inactivated NK-92 NK cells and Jurkat T cells contain a strikingly prominent, novel, ca. 90-kDa tyrosine-phosphorylated protein that we identified as the HSV tegument protein VP11/12. Inasmuch as VP11/12 produced in fibroblasts and epithelial cells is not obviously tyrosine phosphorylated, these data suggested that VP11/12 serves as the substrate of a cell-type-specific protein tyrosine kinase. Consistent with this hypothesis, VP11/12 was also tyrosine phosphorylated in B lymphocytes, and this modification was severely reduced in Jurkat T cells lacking the lymphocyte-specific Src family kinase Lck. These findings demonstrate that HSV tegument proteins can be differentially modified depending on the cell type infected. Our data also raise the possibility that VP11/12 may modulate one or more lymphocyte-specific signaling pathways or serve another lymphocyte specific function. However, HSV type 1 mutants lacking the UL46 gene retained the ability to block signaling through the T-cell receptor in Jurkat cells and remained competent to functionally inactivate the NK-92 NK cell line, indicating that VP11/12 is not essential for lymphocyte inactivation. Further studies are therefore required to determine the biological function of tyrosine phosphorylated VP11/12. PMID- 18417567 TI - Vesicular stomatitis virus oncolysis of T lymphocytes requires cell cycle entry and translation initiation. AB - Vesicular stomatitis virus (VSV) is a candidate oncolytic virus that replicates and induces cell death in cancer cells while sparing normal cells. Although defects in the interferon antiviral response facilitate VSV oncolysis, other host factors, including translational and growth regulatory mechanisms, also appear to influence oncolytic virus activity. We previously demonstrated that VSV infection induces apoptosis in proliferating CD4(+) T lymphocytes from adult T-cell leukemia samples but not in resting T lymphocytes or primary chronic lymphocytic leukemia cells that remain arrested in G(0). Activation of primary CD4(+) T lymphocytes with anti-CD3/CD28 is sufficient to induce VSV replication and cell death in a manner dependent on activation of the MEK1/2, c-Jun NH(2)-terminal kinase, or phosphatidylinositol 3-kinase pathway but not p38. VSV replication is specifically impaired by the cell cycle inhibitor olomoucine or rapamycin, which induces early G(1) arrest, but not by aphidicolin or Taxol, which blocks at the G(1)1S or G(2)1M phase, respectively; this result suggests a requirement for cell cycle entry for efficient VSV replication. The relationship between increased protein translation following G(0)/G(1) transition and VSV permissiveness is highlighted by the absence of mTOR and/or eIF4E phosphorylation whenever VSV replication is impaired. Furthermore, VSV protein production in activated T cells is diminished by small interfering RNA-mediated eIF4E knockdown. These results demonstrate that VSV replication in primary T lymphocytes relies on cell cycle transition from the G(0) phase to the G(1) phase, which is characterized by a sharp increase in ribogenesis and protein synthesis. PMID- 18417568 TI - Characterization of the early steps of infection of primary blood monocytes by human immunodeficiency virus type 1. AB - Blood-circulating monocytes migrate in tissues in response to danger stimuli and differentiate there into two major actors of the immune system: macrophages and dendritic cells. Given their migratory behavior and their pivotal role in the orchestration of immune responses, it is not surprising that cells of the monocyte lineage are the target of several viruses, including human immunodeficiency virus type 1 (HIV-1). HIV-1 replicates in monocytoid cells to an extent that is influenced by their differentiation status and modulated by exogenous stimulations. Unstimulated monocytes display a relative resistance to HIV infection mostly exerted during the early steps of the viral life cycle. Despite intensive studies, the identity of the affected step remains controversial, although it is generally assumed to take place after viral entry. We reexamine here the early steps of viral infection of unstimulated monocytes using vesicular stomatitis virus G protein-pseudotyped HIV-1 virions. Our data indicate that a first block to the early steps of infection of monocytes with these particles occurs at the level of viral entry. After entry, reverse transcription and integration proceed with extremely slow kinetics rather than being blocked. Once completed, viral DNA molecules delay entry into the nucleus and integration for up to 5 to 6 days. The inefficacy of these steps accounts for the resistance of monocytes to HIV-1 during the early steps of infection. PMID- 18417569 TI - Construction and characterization of a human T-cell lymphotropic virus type 3 infectious molecular clone. AB - We and others have uncovered the existence of human T-cell lymphotropic virus type 3 (HTLV-3). We have now generated an HTLV-3 proviral clone. We established that gag, env, pol, pro, and tax/rex as well as minus-strand mRNAs are present in cells transfected with the HTLV-3 clone. HTLV-3 p24(gag) protein is detected in the cell culture supernatant. Transfection of 293T-long terminal repeat (LTR) green fluorescent protein (GFP) cells with the HTLV-3 clone promotes formation of syncytia, a hallmark of Env expression, together with the appearance of fluorescent cells, demonstrating that Tax is expressed. Viral particles are visible by electron microscopy. These particles are infectious, as demonstrated by infection experiments with purified virions. PMID- 18417570 TI - Basolateral entry and release of New and Old World arenaviruses from human airway epithelia. AB - Transmission of arenaviruses from rodent hosts to humans is generally thought to occur through inhalation or ingestion of dust or droplets containing viral particles. Here we demonstrate that two identified arenavirus receptors, alpha dystroglycan (alpha-DG) and transferrin receptor 1 (TfR1), are expressed in polarized human airway epithelia. Lymphocytic choriomeningitis virus strains with high or low alpha-DG affinity and Junin virus, which binds TfR1, efficiently infected polarized epithelia only when applied to the basolateral surface or when injury compromised tight junction integrity. Viral egress from infected epithelia exhibited basolateral polarity. This study demonstrates that respiratory entry of arenaviruses occurs via basolateral receptors. PMID- 18417571 TI - A new role for ns polyprotein cleavage in Sindbis virus replication. AB - One of the distinguishing features of the alphaviruses is a sequential processing of the nonstructural polyproteins P1234 and P123. In the early stages of the infection, the complex of P123+nsP4 forms the primary replication complexes (RCs) that function in negative-strand RNA synthesis. The following processing steps make nsP1+P23+nsP4, and later nsP1+nsP2+nsP3+nsP4. The latter mature complex is active in positive-strand RNA synthesis but can no longer produce negative strands. However, the regulation of negative- and positive-strand RNA synthesis apparently is not the only function of ns polyprotein processing. In this study, we developed Sindbis virus mutants that were incapable of either P23 or P123 cleavage. Both mutants replicated in BHK-21 cells to levels comparable to those of the cleavage-competent virus. They continuously produced negative-strand RNA, but its synthesis was blocked by the translation inhibitor cycloheximide. Thus, after negative-strand synthesis, the ns proteins appeared to irreversibly change conformation and formed mature RCs, in spite of the lack of ns polyprotein cleavage. However, in the cells having no defects in alpha/beta interferon (IFN alpha/beta) production and signaling, the cleavage-deficient viruses induced a high level of type I IFN and were incapable of causing the spread of infection. Moreover, the P123-cleavage-deficient virus was readily eliminated, even from the already infected cells. We speculate that this inability of the viruses with unprocessed polyprotein to productively replicate in the IFN-competent cells and in the cells of mosquito origin was an additional, important factor in ns polyprotein cleavage development. In the case of the Old World alphaviruses, it leads to the release of nsP2 protein, which plays a critical role in inhibiting the cellular antiviral response. PMID- 18417572 TI - Cytomegalovirus proteins vMIA and m38.5 link mitochondrial morphogenesis to Bcl-2 family proteins. AB - Apoptosis is a host defense mechanism against viruses that can be subverted by viral gene products. Human cytomegalovirus encodes viral mitochondria-localized inhibitor of apoptosis (vMIA; also known as pUL37x1), which is targeted to mitochondria and functions as a potent cell death suppressor by binding to and inhibiting proapoptotic Bcl-2 family members Bax and Bak. vMIA expression also dramatically alters mitochondrial morphology, causing the fragmentation of these organelles. A potential ortholog of vMIA, m38.5, which was identified in murine cytomegalovirus, has been shown to localize to mitochondria and protect against chemically induced apoptosis by unknown mechanisms. Despite sharing negligible homology with vMIA and no region detectably corresponding to the vMIA Bax-binding domain, we find that m38.5, like vMIA, binds to Bax and recruits Bax to mitochondria. Interestingly, m38.5 and vMIA appear to block Bax downstream of translocation to mitochondria and after an initial stage of Bax conformational change. In contrast to vMIA, m38.5 neither binds to Bak nor causes mitochondrial fragmentation. Consistently with Bax-selective inactivation by m38.5, m38.5 fragments mitochondria in Bak knockout (KO) cells and protects Bak KO cells from apoptosis better than Bax KO cells. Thus, vMIA and m38.5 share some, but not all, features of apoptosis regulation through Bcl-2 family interaction and allow the dissection of Bax translocation into discrete steps. PMID- 18417573 TI - Henipavirus V protein association with Polo-like kinase reveals functional overlap with STAT1 binding and interferon evasion. AB - Emerging viruses in the paramyxovirus genus Henipavirus evade host antiviral responses via protein interactions between the viral V and W proteins and cellular STAT1 and STAT2 and the cytosolic RNA sensor MDA5. Polo-like kinase (PLK1) is identified as being an additional cellular partner that can bind to Nipah virus P, V, and W proteins. For both Nipah virus and Hendra virus, contact between the V protein and the PLK1 polo box domain is required for V protein phosphorylation. Results indicate that PLK1 is engaged by Nipah virus V protein amino acids 100 to 160, previously identified as being the STAT1 binding domain responsible for host interferon (IFN) signaling evasion, via a Thr-Ser-Ser-Pro motif surrounding residue 130. A distinct Ser-Thr-Pro motif surrounding residue 199 mediates the PLK1 interaction with Hendra virus V protein. Select mutations in the motif surrounding residue 130 also influenced STAT1 binding and innate immune interference, and data indicate that the V:PLK1 and V:STAT complexes are V mediated yet independent of one another. The effects of STAT1/PLK1 binding motif mutations on the function the Nipah virus P protein in directing RNA synthesis were tested. Remarkably, mutations that selectively disrupt the STAT or PLK1 interaction site have no effects on Nipah virus P protein-mediated viral RNA synthesis. Therefore, mutations targeting V protein-mediated IFN evasion will not alter the RNA synthetic capacity of the virus, supporting an attenuation strategy based on disrupting host protein interactions. PMID- 18417574 TI - Coronavirus nonstructural protein 16 is a cap-0 binding enzyme possessing (nucleoside-2'O)-methyltransferase activity. AB - The coronavirus family of positive-strand RNA viruses includes important pathogens of livestock, companion animals, and humans, including the severe acute respiratory syndrome coronavirus that was responsible for a worldwide outbreak in 2003. The unusually complex coronavirus replicase/transcriptase is comprised of 15 or 16 virus-specific subunits that are autoproteolytically derived from two large polyproteins. In line with bioinformatics predictions, we now show that feline coronavirus (FCoV) nonstructural protein 16 (nsp16) possesses an S adenosyl-L-methionine (AdoMet)-dependent RNA (nucleoside-2'O)-methyltransferase (2'O-MTase) activity that is capable of cap-1 formation. Purified recombinant FCoV nsp16 selectively binds to short capped RNAs. Remarkably, an N7-methyl guanosine cap ((7Me)GpppAC(3-6)) is a prerequisite for binding. High-performance liquid chromatography analysis demonstrated that nsp16 mediates methyl transfer from AdoMet to the 2'O position of the first transcribed nucleotide, thus converting (7Me)GpppAC(3-6) into (7Me)GpppA(2')(O)(Me)C(3-6). The characterization of 11 nsp16 mutants supported the previous identification of residues K45, D129, K169, and E202 as the putative K-D-K-E catalytic tetrad of the enzyme. Furthermore, residues Y29 and F173 of FCoV nsp16, which may be the functional counterparts of aromatic residues involved in substrate recognition by the vaccinia virus MTase VP39, were found to be essential for both substrate binding and 2'O-MTase activity. Finally, the weak inhibition profile of different AdoMet analogues indicates that nsp16 has evolved an atypical AdoMet binding site. Our results suggest that coronavirus mRNA carries a cap-1, onto which 2'O methylation follows an order of events in which 2'O-methyl transfer must be preceded by guanine N7 methylation, with the latter step being performed by a yet unknown N7-specific MTase. PMID- 18417575 TI - Primate lentivirus capsid sensitivity to TRIM5 proteins. AB - Mammalian cells express several factors that inhibit lentiviral infection and that have been under strong selective pressure. One of these factors, TRIM5, targets the capsid protein of incoming retrovirus particles and inhibits subsequent steps of the replication cycle. By substituting human immunodeficiency virus type 1 capsid, we were able to show that a set of divergent primate lentivirus capsids was generally not susceptible to restriction by TRIM5 proteins from higher primates. TRIM5alpha proteins from other primates exhibited distinct restriction specificities for primate lentivirus capsids. Finally, we identified novel primate lentiviral capsids that are targeted by TRIMCyp proteins. PMID- 18417576 TI - A conserved sequence within the H2 subunit of the vaccinia virus entry/fusion complex is important for interaction with the A28 subunit and infectivity. AB - The recently described vaccinia virus entry/fusion complex (EFC) comprises at least eight polypeptides that are conserved in all poxviruses. Neither the structure of the complex nor the roles of individual subunits are known. Here we provide evidence for an interaction between the H2 and A28 subunits in the context of a virus infection as well as in uninfected cells transfected with plasmids expressing the corresponding genes. We focused on a highly conserved 21 amino acid-segment in H2 that is flanked by cysteine residues. The effect of amino acid substitutions within the 21-amino-acid segment was determined by an infectivity complementation assay using a conditional H2-null mutant of vaccinia virus. Mutations that had no, moderate, or large negative effects on complementation were found. The latter group included glutamic acid substitutions of leucine and individual glycines and alanine substitution of both glycines within a LGYSG sequence. Mutations with the most pronounced effect on infectivity disrupted the interaction of H2 with A28 to the greatest extent in both infected and uninfected cells. These data indicate that the LGYSG sequence is important for the interaction of H2 with A28 and suggest that this sequence is buried within the EFC complex. PMID- 18417577 TI - Identification of a physiological phosphorylation site of the herpes simplex virus 1-encoded protein kinase Us3 which regulates its optimal catalytic activity in vitro and influences its function in infected cells. AB - Us3 is a serine/threonine protein kinase encoded by herpes simplex virus 1 (HSV 1). Here, we report the identification of a physiological Us3 phosphorylation site on serine at position 147 (Ser-147) which regulates its protein kinase activity in vitro. Moreover, mutation of this site influences Us3 function, including correct localization of the enzyme and induction of the usual morphological changes in HSV-1-infected cells. These conclusions are based on the following observations: (i) in in vitro kinase assays, a domain of Us3 containing Ser-147 was specifically phosphorylated by Us3 and protein kinase A, while a mutant domain in which Ser-147 was replaced with alanine was not; (ii) in vitro, alanine replacement of Ser-147 (S147A) in Us3 resulted in significant impairment of the kinase activity of the purified molecule expressed in a baculovirus system; (iii) phosphorylation of Ser-147 in Us3 tagged with the monomeric fluorescent protein (FP) VenusA206K (VenusA206K-Us3) from Vero cells infected with a recombinant HSV-1 encoding VenusA206K-Us3 was specifically detected using an antibody that recognizes phosphorylated serine or threonine residues with arginine at the -3 and -2 positions; and (iv) the S147A mutation influenced some but not all Us3 functions, including the ability of the protein to localize itself properly and to induce wild-type cytopathic effects in infected cells. Our results suggest that some of the regulatory activities of Us3 in infected cells are controlled by phosphorylation at Ser-147. PMID- 18417578 TI - Interferon regulatory factor 4 is involved in Epstein-Barr virus-mediated transformation of human B lymphocytes. AB - Epstein-Barr virus (EBV) infection is associated with many human malignancies. In vitro, EBV transforms primary B lymphocytes into continuously growing lymphoblastoid cell lines. EBV latent membrane protein 1 (LMP-1) is required for EBV transformation processes. Interferon regulatory factor 4 (IRF-4) is a transcription factor and has oncogenic potential. We find that high levels of IRF 4 are associated with EBV transformation of human primary B cells in vitro and with EBV type III latency in which LMP-1 is expressed. We show that EBV LMP-1 stimulates IRF-4 expression in B lymphocytes. The stimulation of IRF-4 by LMP-1 requires signaling from LMP-1 and involves cellular NF-kappaB. The growth of EBV transformed cells is inhibited when IRF-4 is specifically down-regulated. We further demonstrate that IRF-4 knockdown cells have lower proliferation but higher apoptotic rates than control cells. Finally, IRF-4 is expressed in significant numbers of specimens of primary central nervous system (CNS) lymphomas (12/27 [44.4%]), an EBV-associated malignancy. The association between the expression levels of LMP-1 and IRF-4 is statistically significant (P = 0.011) in these CNS lymphomas. Our data suggest that IRF-4 may be a critical factor in EBV transformation and a useful target in the therapy of EBV-mediated neoplasia. PMID- 18417579 TI - Antibody is critical for the clearance of murine norovirus infection. AB - Human noroviruses cause more than 90% of epidemic nonbacterial gastroenteritis. However, the role of B cells and antibody in the immune response to noroviruses is unclear. Previous studies have demonstrated that human norovirus specific antibody levels increase upon infection, but they may not be protective against infection. In this report, we used murine norovirus (MNV), an enteric norovirus, as a model to determine the importance of norovirus specific B cells and immune antibody in clearance of norovirus infection. We show here that mice genetically deficient in B cells failed to clear primary MNV infection as effectively as wild type mice. In addition, adoptively transferred immune splenocytes derived from B cell-deficient mice or antibody production-deficient mice were unable to efficiently clear persistent MNV infection in RAG1(-/-) mice. Further, adoptive transfer of either polyclonal anti-MNV serum or neutralizing anti-MNV monoclonal antibodies was sufficient to reduce the level of MNV infection both systemically and in the intestine. Together, these data demonstrate that antibody plays an important role in the clearance of MNV and that immunoglobulin G anti-norovirus antibody can play an important role in clearing mucosal infection. PMID- 18417580 TI - Novel postentry resistance to AKV ecotropic mouse gammaretroviruses in the African pygmy mouse, Mus minutoides. AB - Cells of Mus minutoides, an African pygmy mouse of the subgenus Nannomys, are susceptible to ecotropic Moloney and Friend mouse leukemia viruses (MLVs) but not to AKV-type MLVs. Transfected MA139 ferret cells expressing the mCAT-1 cell surface receptor, with the minCAT-1 substitutions K222Q and V233L, did not restrict AKV MLV. The resistance of M. minutoides cells to AKV MLV was not relieved by inhibitors of glycosylation or by the introduction of NIH 3T3 mCAT-1. Resistance is thus not mediated by receptor sequence variation, expression level, or glycosylation. M. minutoides cells are also infectible with LacZ pseudotypes having AKV Env and Moloney MLV (MoMLV) Gag proteins, further indicating that AKV Env sequence variations do not contribute to the observed block. The pattern of virus resistance in M. minutoides differs from that of the known variants of the Fv1 postentry resistance gene; M. minutoides is equally resistant to N-, B-, and NR-tropic AKV viruses and is equally susceptible to NR- and NB-tropic Friend MLVs. This novel resistance blocks replication before reverse transcription, whereas Fv1 generally restricts replication after reverse transcription; M. minutoides cells produce 2-long-terminal-repeat viral DNA circles and linear viral DNA after infection with MoMLV but not with AKV MLV. Analysis of MoMLV-AKV MLV chimeras determined that the target of resistance is in the virus capsid gene. Mutagenesis demonstrated that restriction is mediated by two amino acid substitutions, H117L and A110R; substitutions at these sites can also be targeted by the resistance genes Fv1 and TRIM5alpha. M. minutoides cells thus have a novel postentry resistance to AKV MLVs. PMID- 18417581 TI - Priming of CD8+ T cells during central nervous system infection with a murine coronavirus is strain dependent. AB - Virus-specific CD8(+) T cells are critical for protection against neurotropic coronaviruses; however, central nervous system (CNS) infection with the recombinant JHM (RJHM) strain of mouse hepatitis virus (MHV) elicits a weak CD8(+) T-cell response in the brain and causes lethal encephalomyelitis. An adoptive transfer model was used to elucidate the kinetics of CD8(+) T-cell priming during CNS infection with RJHM as well as with two MHV strains that induce a robust CD8(+) T-cell response (RA59 and SJHM/RA59, a recombinant A59 virus expressing the JHM spike). While RA59 and SJHM/RA59 infections resulted in CD8(+) T-cell priming within the first 2 days postinfection, RJHM infection did not lead to proliferation of naive CD8(+) T cells. While all three viruses replicated efficiently in the brain, only RA59 and SJHM/RA59 replicated to appreciable levels in the cervical lymph nodes (CLN), the site of T-cell priming during acute CNS infection. RJHM was unable to suppress the CD8(+) T-cell response elicited by RA59 in mice simultaneously infected with both strains, suggesting that RJHM does not cause generalized immunosuppression. RJHM was also unable to elicit a secondary CD8(+) T-cell response in the brain following peripheral immunization against a viral epitope. Notably, the weak CD8(+) T-cell response elicited by RJHM was unique to CNS infection, since peripheral inoculation induced a robust CD8(+) T-cell response in the spleen. These findings suggest that the failure of RJHM to prime a robust CD8(+) T-cell response during CNS infection is likely due to its failure to replicate in the CLN. PMID- 18417582 TI - A single Banana streak virus integration event in the banana genome as the origin of infectious endogenous pararetrovirus. AB - Sequencing of plant nuclear genomes reveals the widespread presence of integrated viral sequences known as endogenous pararetroviruses (EPRVs). Banana is one of the three plant species known to harbor infectious EPRVs. Musa balbisiana carries integrated copies of Banana streak virus (BSV), which are infectious by releasing virions in interspecific hybrids. Here, we analyze the organization of the EPRV of BSV Goldfinger (BSGfV) present in the wild diploid M. balbisiana cv. Pisang Klutuk Wulung (PKW) revealed by the study of Musa bacterial artificial chromosome resources and interspecific genetic cross. cv. PKW contains two similar EPRVs of BSGfV. Genotyping of these integrants and studies of their segregation pattern show an allelic insertion. Despite the fact that integrated BSGfV has undergone extensive rearrangement, both EPRVs contain the full-length viral genome. The high degree of sequence conservation between the integrated and episomal form of the virus indicates a recent integration event; however, only one allele is infectious. Analysis of BSGfV EPRV segregation among an F1 population from an interspecific genetic cross revealed that these EPRV sequences correspond to two alleles originating from a single integration event. We describe here for the first time the full genomic and genetic organization of the two EPRVs of BSGfV present in cv. PKW in response to the challenge facing both scientists and breeders to identify and generate genetic resources free from BSV. We discuss the consequences of this unique host-pathogen interaction in terms of genetic and genomic plant defenses versus strategies of infectious BSGfV EPRVs. PMID- 18417583 TI - Infection with Vpr-positive human immunodeficiency virus type 1 impairs NK cell function indirectly through cytokine dysregulation of infected target cells. AB - Human immunodeficiency virus type 1 (HIV-1) infection has been implicated in impairing various aspects of NK cell function in viremic condition, and several viral factors contribute to these defects. Here, we evaluated the effect of HIV-1 Vpr on NK cell cytolytic function and cytokine (gamma interferon [IFN-gamma]) production in the context of infection and exposure. Our data indicate that NK cells derived from a peripheral blood mononuclear cell culture infected in vitro with HIV-1 vpr(+) virus or exposed to recombinant Vpr protein exhibited reduced target cell killing in conjunction with diminished expression of CD107a and reduced IFN-gamma production compared to their Vpr-negative counterparts. This Vpr-induced NK cell defect is in part through differential regulation of interleukin-12 and transforming growth factor beta production by the infected target cells and concomitant activation of Smad3 signaling pathway. Collectively, these results illustrate the ability of Vpr to impair NK cell-mediated innate immune functions indirectly by dysregulating multiple cytokines in the infected target cells, thus increasing disease severity and affecting the final outcome in HIV-1 infection. PMID- 18417584 TI - Identification of a critical motif for the human immunodeficiency virus type 1 (HIV-1) gp41 core structure: implications for designing novel anti-HIV fusion inhibitors. AB - Human immunodeficiency virus type 1 (HIV-1) entry into the host cell involves a cascade of events and currently represents one of most attractive targets in the search for new antiviral drugs. The fusion-active gp41 core structure is a stable six-helix bundle (6-HB) folded by its trimeric N-terminal heptad repeat (NHR) and C-terminal heptad repeat (CHR). Peptides derived from the CHR region of HIV-1 gp41 are potent fusion inhibitors that target the NHR to block viral and cellular membrane fusion in a dominant negative fashion. However, all CHR peptides reported to date are derived primarily from residues 628 to 673 of gp41; little attention has been paid to the upstream sequence of the pocket binding domain (PBD) in the CHR. Here, we have identified a motif ((621)QIWNNMT(627)) located at the upstream region of the gp41 CHR, immediately adjacent to the PBD ((628)WMEWEREI(635)). Biophysical characterization demonstrated that this motif is critical for the stabilization of the gp41 6-HB core. The peptide CP621-652, containing the (621)QIWNNMT(627) motif, was able to interact with T21, a counterpart peptide derived from the NHR, to form a typical 6-HB structure with a high thermostability (thermal unfolding transition [T(m)] value of 82 degrees C). In contrast, the 6-HB formed by the peptides N36 and C34, which has been considered to be a core structure of the fusion-active gp41, had a T(m) of 64 degrees C. Different from T-20 (brand name Fuseon), which is the first and only HIV-1 fusion inhibitor approved for clinical use, CP621-652 could efficiently block 6-HB formation in a dose-dependent manner. Significantly, CP621-652 had potent inhibitory activity against HIV-1-mediated cell-cell fusion and infection, especially against T-20- and C34-resistant virus. Therefore, our works provide important information for understanding the core structure of the fusion-active gp41 and for designing novel anti-HIV peptides. PMID- 18417585 TI - Amino acids 143 to 150 of the herpes simplex virus type 1 scaffold protein are required for the formation of portal-containing capsids. AB - The herpes simplex virus type 1 (HSV-1) portal is composed of a dodecamer of UL6 protein molecules whose incorporation into the capsid is mediated by interaction with the HSV-1 UL26.5 scaffold protein. Previous results with an in vitro capsid assembly assay demonstrated that nine amino acids (amino acids 143 to 151) of the UL26.5 protein are required for its interaction with UL6 and for incorporation of the portal complex into capsids. In the present study an HSV-1 mutant, bvFH411, was isolated and contained a deletion that removed the codons for UL26.5 amino acids 143 to 150. The mutant virus failed to produce infectious virus in noncomplementing cells, and only B capsids that contained only minor amounts of portal protein were made. These data corroborate our previous in vitro studies and demonstrate that amino acids 143 to 150 of UL26.5 are required for the formation of portal-containing HSV-1 capsids. PMID- 18417586 TI - Rapid sequence change and geographical spread of human parvovirus B19: comparison of B19 virus evolution in acute and persistent infections. AB - Parvovirus B19 is a common human pathogen maintained by horizontal transmission between acutely infected individuals. However, B19 virus can also be detected in tissues throughout the life of the host, although little is understood about the nature of such persistence. In the current study, we created large VP1/2 sequence data sets of plasma- and tissue (autopsy)-derived variants of B19 virus with known sample dates to compare the rates of sequence change in exogenous virus populations with those in persistently infected individuals. By using linear regression and likelihood-based methods (such as the BEAST program), we found that plasma-derived B19 virus showed a substitution rate of 4 x 10(-4) and an unconstrained (synonymous)-substitution rate of 18 x 10(-4) per site per year, several times higher than previously estimated and within the range of values for mammalian RNA viruses. The underlying high mutation frequency implied by these substitution rates may enable rapid adaptive changes that are more commonly ascribed to RNA virus populations. These revised estimates predict that the last common ancestor for currently circulating genotype 1 variants of B19 virus existed around 1956 to 1959, fitting well with previous analyses of the B19 virus "bioportfolio" that support a complete cessation of genotype 2 infections and their replacement by genotype 1 infections in the 1960s. In contrast, the evolution of B19 virus amplified from tissue samples was best modeled by using estimated dates of primary infection rather than sample dates, consistent with slow or absent sequence change during persistence. Determining what epidemiological or biological factors led to such a complete and geographically extensive population replacement over this short period is central to further understanding the nature of parvovirus evolution. PMID- 18417587 TI - Sulfatide is required for efficient replication of influenza A virus. AB - Sulfatide is abundantly expressed in various mammalian organs, including the intestines and trachea, in which influenza A viruses (IAVs) replicate. However, the function of sulfatide in IAV infection remains unknown. Sulfatide is synthesized by two transferases, ceramide galactosyltransferase (CGT) and cerebroside sulfotransferase (CST), and is degraded by arylsulfatase A (ASA). In this study, we demonstrated that sulfatide enhanced IAV replication through efficient translocation of the newly synthesized IAV nucleoprotein (NP) from the nucleus to the cytoplasm, by using genetically produced cells in which sulfatide expression was down-regulated by RNA interference against CST mRNA or overexpression of the ASA gene and in which sulfatide expression was up-regulated by overexpression of both the CST and CGT genes. Treatment of IAV-infected cells with an antisulfatide monoclonal antibody (MAb) or an anti-hemagglutinin (HA) MAb, which blocks the binding of IAV and sulfatide, resulted in a significant reduction in IAV replication and accumulation of the viral NP in the nucleus. Furthermore, antisulfatide MAb protected mice against lethal challenge with pathogenic influenza A/WSN/33 (H1N1) virus. These results indicate that association of sulfatide with HA delivered to the cell surface induces translocation of the newly synthesized IAV ribonucleoprotein complexes from the nucleus to the cytoplasm. Our findings provide new insights into IAV replication and suggest new therapeutic strategies. PMID- 18417588 TI - A filovirus-unique region of Ebola virus nucleoprotein confers aberrant migration and mediates its incorporation into virions. AB - The Ebola virus nucleoprotein (NP) is an essential component of the nucleocapsid, required for filovirus particle formation and replication. Together with virion protein 35 (VP35) and VP24, this gene product gives rise to the filamentous nucleocapsid within transfected cells. Ebola virus NP migrates aberrantly, with an apparent molecular mass of 115 kDa, although it is predicted to encode an approximately 85-kDa protein. In this report, we show that two domains of this protein determine this aberrant migration and that this region mediates its incorporation into virions. These regions, amino acids 439 to 492 and amino acids 589 to 739, alter the mobility of Ebola virus NP by sodium dodecyl sulfate polyacrylamide gel electrophoresis by 5 and 15 kDa, respectively, and confer similar effects on a heterologous protein, LacZ, in a position-independent fashion. Furthermore, when coexpressed with VP40, VP35, and VP24, this region mediated incorporation of NP into released viruslike particles. When fused to chimeric paramyxovirus NPs derived from measles or respiratory syncytial virus, this domain directed these proteins into the viruslike particle. The COOH terminal NP domain comprises a conserved highly acidic region of NP with predicted disorder, distinguishing Ebola virus NPs from paramyxovirus NPs. The acidic character of this domain is likely responsible for its aberrant biochemical properties. These findings demonstrate that this region is essential for the assembly of the filamentous nucleocapsids that give rise to filoviruses. PMID- 18417589 TI - Laboratory strains of murine cytomegalovirus are genetically similar to but phenotypically distinct from wild strains of virus. AB - Murine cytomegalovirus (MCMV) is widely used to model human cytomegalovirus (HCMV) infection. However, it is known that serially passaged laboratory strains of HCMV differ significantly from recently isolated clinical strains of HCMV. It is therefore axiomatic that clinical models of HCMV using serially passaged strains of MCMV may not be able to fully represent the complexities of the system they are attempting to model and may not fully represent the complex biology of MCMV. To determine whether genotypic and phenotypic differences also exist between laboratory strains of MCMV and wild derived strains of MCMV, we sequenced the genomes of three low-passage strains of MCMV, plus the laboratory strain, K181. We coupled this genetic characterization to their phenotypic characteristics. In contrast to what is seen with HCMV (and rhesus CMV), there were no major genomic rearrangements in the MCMV genomes. In addition, the genome size was remarkably conserved between MCMV strains with no major insertions or deletions. There was, however, significant sequence variation between strains of MCMV, particularly at the genomic termini. These more subtle genetic differences led to considerable differences in in vivo replication with some strains of MCMV, such as WP15B, replicating preferentially in otherwise-MCMV-resistant C57BL/6 mice. CBA mice were no more resistant to MCMV than C57BL/6 mice and for some MCMV strains appeared to control infection less well than C57BL/6 mice. It is apparent that the previously described host resistance patterns of inbred mice and MCMV are not consistently applicable for all MCMV strains. PMID- 18417590 TI - Vaccinia virus DNA ligase recruits cellular topoisomerase II to sites of viral replication and assembly. AB - Vaccinia virus replication is inhibited by etoposide and mitoxantrone even though poxviruses do not encode the type II topoisomerases that are the specific targets of these drugs. Furthermore, one can isolate drug-resistant virus carrying mutations in the viral DNA ligase and yet the ligase is not known to exhibit sensitivity to these drugs. A yeast two-hybrid screen was used to search for proteins binding to vaccinia ligase, and one of the nine proteins identified comprised a portion (residue 901 to end) of human topoisomerase IIbeta. One can prevent the interaction by introducing a C(11)-to-Y substitution mutation into the N terminus of the ligase bait protein, which is one of the mutations conferring etoposide and mitoxantrone resistance. Coimmunoprecipitation methods showed that the native ligase and a Flag-tagged recombinant protein form complexes with human topoisomerase IIalpha/beta in infected cells and that this interaction can also be disrupted by mutations in the A50R (ligase) gene. Immunofluorescence microscopy showed that both topoisomerase IIalpha and IIbeta antigens are recruited to cytoplasmic sites of virus replication and that less topoisomerase was recruited to these sites in cells infected with mutant virus than in cells infected with wild-type virus. Immunoelectron microscopy confirmed the presence of topoisomerases IIalpha/beta in virosomes, but the enzyme could not be detected in mature virus particles. We propose that the genetics of etoposide and mitoxantrone resistance can be explained by vaccinia ligase binding to cellular topoisomerase II and recruiting this nuclear enzyme to sites of virus biogenesis. Although other nuclear DNA binding proteins have been detected in virosomes, this appears to be the first demonstration of an enzyme being selectively recruited to sites of poxvirus DNA synthesis and assembly. PMID- 18417591 TI - Human parainfluenza virus type 2 V protein inhibits genome replication by binding to the L protein: possible role in promoting viral fitness. AB - The human parainfluenza virus type 2 (hPIV2) V protein plays important roles in inhibiting the host interferon response and promoting virus growth, but its role in hPIV2 replication and transcription is not clear. A green fluorescent protein (GFP)-expressing a negative-sense minigenomic construct of hPIV2 has been established by standard technology, with helper plasmids expressing the nucleocapsid protein (NP), phosphoprotein (P), and large RNA polymerase (L) protein, to examine the role of V protein. We found that the simultaneous expression of wild-type V protein in the minigenome system inhibited GFP expression, at least in part, by inhibiting minigenome replication. In contrast, expression of C terminally truncated or mutant hPIV2 V proteins had no effect. Moreover, the V protein of simian virus 41, the rubulavirus most closely related virus to hPIV2, also inhibited GFP expression, whereas that of PIV5, a more distantly related rubulavirus, did not. Using these other rubulavirus V proteins, as well as various mutant hPIV2 V proteins, we found that the ability of V protein to inhibit GFP expression correlated with its ability to bind to L protein via its C-terminal V protein-specific region, but there was no correlation with NP binding. A possible role for this inhibition of genome replication in promoting viral fitness is discussed. PMID- 18417592 TI - Porcine rotavirus bearing an aberrant gene stemming from an intergenic recombination of the NSP2 and NSP5 genes is defective and interfering. AB - Serial undiluted passage of a porcine rotavirus in MA104 cells yielded three distinct virus populations, each of which bore different rearranged genes. Sequencing revealed that each of two populations bore a distinct intragenic recombinant NSP3 gene consisting of a partial duplication in a head-to-tail orientation without altering the NSP3 open reading frame and the third population carried both an intragenic recombinant NSP3 gene and an intergenic recombinant gene (1,647 nucleotides in length) which contained a truncated NSP2 gene inserted into the NSP5 gene at residue 332. The former two populations were viable, whereas the latter population was defective and interfering. PMID- 18417593 TI - Length requirements for membrane fusion of influenza virus hemagglutinin peptide linkers to transmembrane or fusion peptide domains. AB - During membrane fusion, the influenza A virus hemagglutinin (HA) adopts an extended helical structure that contains the viral transmembrane and fusion peptide domains at the same end of the molecule. The peptide segments that link the end of this rod-like structure to the membrane-associating domains are approximately 10 amino acids in each case, and their structure at the pH of fusion is currently unknown. Here, we examine mutant HAs and influenza viruses containing such HAs to determine whether these peptide linkers are subject to specific length requirements for the proper folding of native HA and for membrane fusion function. Using pairwise deletions and insertions, we show that the region flanking the fusion peptide appears to be important for the folding of the native HA structure but that mutant proteins with small insertions can be expressed on the cell surface and are functional for membrane fusion. HA mutants with deletions of up to 10 residues and insertions of as many as 12 amino acids were generated for the peptide linker to the viral transmembrane domain, and all folded properly and were expressed on the cell surface. For these mutants, it was possible to designate length restrictions for efficient membrane fusion, as functional activity was observed only for mutants containing linkers with insertions or deletions of eight residues or less. The linker peptide mutants are discussed with respect to requirements for the folding of native HAs and length restrictions for membrane fusion activity. PMID- 18417594 TI - Authentic replication and recombination of Tomato bushy stunt virus RNA in a cell free extract from yeast. AB - To study the replication of Tomato bushy stunt virus (TBSV), a small tombusvirus of plants, we have developed a cell-free system based on a Saccharomyces cerevisiae extract. The cell-free system was capable of performing a complete replication cycle on added plus-stranded TBSV replicon RNA (repRNA) that led to the production of approximately 30-fold-more plus-stranded progeny RNAs than the minus-stranded replication intermediate. The cell-free system also replicated the full-length TBSV genomic RNA, which resulted in production of subgenomic RNAs as well. The cell-free system showed high template specificity, since a mutated repRNA, minus-stranded repRNA, or a heterologous viral RNA could not be used as templates by the tombusvirus replicase. Similar to the in vivo situation, replication of the TBSV replicon RNA took place in a membraneous fraction, in which the viral replicase-RNA complex was RNase and protease resistant but sensitive to detergents. In addition to faithfully replicating the TBSV replicon RNA, the cell-free system was also capable of generating TBSV RNA recombinants with high efficiency. Altogether, tombusvirus replicase in the cell-free system showed features remarkably similar to those of the in vivo replicase, including carrying out a complete cycle of replication, high template specificity, and the ability to recombine efficiently. PMID- 18417595 TI - Sindbis virus conformational changes induced by a neutralizing anti-E1 monoclonal antibody. AB - A rare Sindbis virus anti-E1 neutralizing monoclonal antibody, Sin-33, was investigated to determine the mechanism of in vitro neutralization. A cryoelectron microscopic reconstruction of Sindbis virus (SVHR) neutralized with FAb from Sin-33 (FAb-33) revealed conformational changes on the surface of the virion at a resolution of 24 A. FAb-33 was found to bind E1 in less than 1:1 molar ratios, as shown by the absence of FAb density in the reconstruction and stoichiometric measurements using radiolabeled FAb-33, which determined that about 60 molecules of FAb-33 bound to the 240 possible sites in a single virus particle. FAb-33-neutralized virus particles became sensitive to digestion by endoproteinase Glu-C, providing further evidence of antibody-induced structural changes within the virus particle. The treatment of FAb-33-neutralized or Sin-33 neutralized SVHR with low pH did not induce the conformational rearrangements required for virus membrane-cell membrane fusion. Exposure to low pH, however, increased the amount of Sin-33 or FAb-33 that bound to the virus particles, indicating the exposure of additional epitopes. The neutralization of SVHR infection by FAb-33 or Sin-33 did not prevent the association of virus with host cells. These data are in agreement with the results of previous studies that demonstrated that specific antibodies can inactivate the infectious state of a metastable virus in vitro by the induction of conformational changes to produce an inactive structure. A model is proposed which postulates that the induction of conformational changes in the infectious state of a metastable enveloped virus may be a general mechanism of antibody inactivation of virus infectivity. PMID- 18417596 TI - Bovine papillomavirus type 1: from clathrin to caveolin. AB - Viruses may infect cells through clathrin-dependent, caveolin-dependent, or clathrin- and caveolin-independent endocytosis. Bovine papillomavirus type 1 (BPV1) entry into cells has been shown to occur by clathrin-dependent endocytosis, a pathway that involves the formation of clathrin-coated pits and fusion to early endosomes. Recently, it has been demonstrated that the closely related JC virus can enter cells in clathrin-coated vesicles and subsequently traffic to caveolae, the organelle where vesicles of the caveolin-dependent pathway deliver their cargo. In this study, we use immunofluorescence staining of BPV1 pseudovirions to show that BPV1 overlaps with the endosome marker EEA1 early during infection and later colocalizes with caveolin-1. We provide evidence through the colocalization of BPV1 with transferrin and cholera toxin B that BPVl trafficking may not be restricted to the clathrin-dependent pathway. Disrupting the entry of caveolar vesicles did not affect BPV1 infection; however, we show that blocking the caveolar pathway postentry results in a loss of BPV1 infection. These data indicate that BPV1 may enter by clathrin-mediated endocytosis and then utilize the caveolar pathway for infection, a pattern of trafficking that may explain the slow kinetics of BPV1 infection. PMID- 18417598 TI - Comparison of echo-planar diffusion-weighted imaging and delayed postcontrast T1 weighted MR imaging for the detection of residual cholesteatoma. AB - BACKGROUND AND PURPOSE: Echo-planar diffusion-weighted imaging (DWI) and delayed postcontrast T1-weighted MR imaging (DPI) have been proposed in previous studies to detect residual middle ear cholesteatomas, with varying results. We assessed and compared these 2 techniques in patients with canal wall-up tympanoplasty. MATERIALS AND METHODS: This was a prospective cohort study. Patients who underwent surgery for middle ear cholesteatoma had CT scanning 9 months after the surgery. If opacity was observed (64%) on CT scans, DWI and DPI were performed before second-look surgery. CT, MR imaging, and surgical data were available for 31 patients. Charts were reviewed independently by 3 blinded examiners. Interobserver agreement for MR imaging was calculated (Cohen kappa). Sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) were calculated for these techniques: 1) alone or in association, and 2) according to the residual cholesteatoma size measured during surgery. RESULTS: Interobserver agreement was better for DWI (kappa = 0.81) than for DPI (kappa = 0.51). Sensitivity, specificity, PPV, and NPV values were 60%, 72.73%, 80%, and 50%, respectively, with DWI; and 90%, 54.55%, 78.26%, and 75%, respectively, with DPI. With cholesteatomas >5 mm, the sensitivity and specificity of DWI reached 100% and 88%, respectively, with values for DPI reaching 100% and 80%, respectively. The association of both techniques only allowed improvements in the specificity for lesions >5 mm. CONCLUSIONS: Both techniques gave acceptable results for residual cholesteatoma detection. DWI is more specific but less sensitive than DPI. Their concurrent use may benefit patients by avoiding undue surgery. PMID- 18417597 TI - Hepatitis C virus (HCV)-induced immunoglobulin hypermutation reduces the affinity and neutralizing activities of antibodies against HCV envelope protein. AB - Hepatitis C virus (HCV) often causes persistent infection despite the presence of neutralizing antibodies against the virus in the sera of hepatitis C patients. HCV infects both hepatocytes and B cells through the binding of its envelope glycoprotein E2 to CD81, the putative viral receptor. Previously, we have shown that E2-CD81 interaction induces hypermutation of heavy-chain immunoglobulin (V(H)) in B cells. We hypothesize that if HCV infects antibody-producing B cells, the resultant hypermutation of V(H) may lower the affinity and specificity of the HCV-specific antibodies, enabling HCV to escape from immune surveillance. To test this hypothesis, we infected human hybridoma clones producing either neutralizing or non-neutralizing anti-E2 or anti-E1 antibodies with a lymphotropic HCV (SB strain). All of the hybridoma clones, except for a neutralizing antibody producing hybridoma, could be infected with HCV and support virus replication for at least 8 weeks after infection. The V(H) sequences in the infected hybridomas had a significantly higher mutation frequency than those in the uninfected hybridomas, with mutations concentrating in complementarity-determining region 3. These mutations lowered the antibody affinity against the targeting protein and also lowered the virus-neutralizing activity of anti-E2 antibodies. Furthermore, antibody-mediated complement-dependent cytotoxicity with the antibodies secreted from the HCV-infected hybridomas was impaired. These results suggest that HCV infection could cause some anti-HCV-antibody-producing hybridoma B cells to make less-protective antibodies. PMID- 18417599 TI - Olfactory anomalies in CHARGE syndrome: imaging findings of a potential major diagnostic criterion. AB - BACKGROUND AND PURPOSE: CHARGE syndrome is a genetic disorder resulting in the association of multiple congenital anomalies. Although a high prevalence of olfactory anomalies in CHARGE syndrome has been reported in autopsy and functional studies, to our knowledge, such anomalies have not been included among the diagnostic criteria, and their radiographic prevalence has not been assessed. The purpose of this research was to determine the radiographic prevalence of olfactory anomalies in a small sample of subjects with diagnosed CHARGE syndrome. MATERIALS AND METHODS: The medical records and high-resolution MR images (section thickness < or =3 mm and in-plane resolution < or =1 mm) in 10 patients with clinically proved CHARGE syndrome were retrospectively reviewed by 3 neuroradiologists who consensually evaluated the status of the olfactory bulbs and sulci as either normal, hypoplastic, or absent. The prevalence (p) of congenital anomalies found in the medical records and of the olfactory structures was calculated with a 95% confidence interval (CI). RESULTS: MR imaging demonstrated olfactory anomalies in all 10 patients, including either absence or hypoplasia of the olfactory bulbs and olfactory sulci (p, 100%; CI, 0.65-1.00). CONCLUSION: These findings suggest that olfactory abnormalities detectable on high-resolution MR imaging are among the most prevalent features of CHARGE syndrome. PMID- 18417600 TI - T1 signal intensity and height of the anterior pituitary in neonates: correlation with postnatal time. AB - BACKGROUND AND PURPOSE: The anterior pituitary of a term neonate is usually hyperintense on T1-weighted MR images, which may represent histologic changes of the gland due to the effect of high estrogen levels during the fetal period; however, MR findings of a preterm neonate have not been fully evaluated. The purpose of this study was to investigate whether intensity and size of the neonatal anterior pituitary on MR images obtained near term of corrected age correlates with the gestational age at birth or postnatal time. MATERIALS AND METHODS: Data of 88 consecutive neonates (gestational age, 24-41 weeks; mean, 31.5 weeks) were analyzed. All of the neonates underwent MR imaging at a corrected age of 0 months +/- 4 weeks. Relative signal intensity of the anterior pituitary compared with that of the pons on T1-weighted sagittal images was calculated. Height of the pituitary was also measured. Stepwise regression analysis was performed to evaluate the effects of gestational age at birth and postnatal time on the relative signal intensity and on the pituitary height. RESULTS: The relative signal intensity significantly negatively correlated with postnatal time (P = .001) but not with gestational age at birth (P = .42). Pituitary height significantly negatively correlated with postnatal time (P = .049) but not with gestational age at birth (P = .071). CONCLUSION: A significant negative correlation exists between postnatal time and signal intensity on T1 weighted MR images of the anterior pituitary obtained near term. A nonhyperintense anterior pituitary is a normal MR finding of preterm neonates when imaged near term. PMID- 18417601 TI - Large arachnoid granulations involving the dorsal superior sagittal sinus: findings on MR imaging and MR venography. AB - BACKGROUND AND PURPOSE: Large arachnoid granulations (AG) within the dorsal superior sagittal sinus (SSS) have been incompletely characterized and can be confused with pathology. This report reviews the characteristics of these anatomic structures to establish common imaging features that allow differentiation from pathology. MATERIALS AND METHODS: Twelve cases of large AG in the dorsal SSS are presented, identified by MR imaging. Signal intensity characteristics, size, location, venographic appearance, and association with adjacent venous and osseous structures were documented. RESULTS: A defect in the dura of the SSS was seen in all of the cases communicating with the subjacent subarachnoid space. The average size of the AG was 8.1 x 9.4 x 10.0 mm (range, 4 19 mm). Ten produced calvarial remodeling, and 11 were in the direct vicinity of the lambda. On T2-weighted images, all were hyperintense to the brain. On T1 weighted images, 8 were hypointense and 4 were hypointense with mixed areas of isointense signal intensity. All of the AGs were associated with cortical venous structures entering the sinus. On MR venography, AGs appeared as focal protrusions into the sinus, displacing, distorting, and narrowing the sinus lumen. Seven patients had headache without other visible cause on MR imaging, and 4 were initially interpreted as thrombosis or tumor. CONCLUSION: Large AGs can occur in the dorsal SSS. They are well-defined projections of the subarachnoid space into the sinus, can cause luminal narrowing and calvarial remodeling, and have typical signal intensity characteristics, position, and morphology differentiating them from other pathology. Association with patient symptoms is uncertain. PMID- 18417602 TI - Carotid body tumors: objective criteria to predict the Shamblin group on MR imaging. AB - BACKGROUND AND PURPOSE: MR imaging is an established method for diagnosis and surgical planning of carotid body tumors (CBTs). However no studies have elaborated preoperative objective criteria to predict the Shamblin (surgical) classification of CBTs, an important predictor of vascular morbidity. The purpose of this study was to establish criteria to accurately predict the Shamblin group on preoperative MR imaging for a uniform reporting system. MATERIALS AND METHODS: MR images of 9 CBTs in 8 consecutive patients who underwent surgery between 2004 and 2007 were reviewed at a tertiary cancer hospital. The surgical records were blinded to the radiologists. A radiologic classification into 3 types (I, II, and III) based on the maximum degree of circumferential contact of the tumor with the internal carotid artery (ICA) was attempted and correlated with the Shamblin group in surgical records. RESULTS: There were 5 type III, 3 type II, and 1 type I tumors. The type I tumor had an ICA maximum circumference of contact less than or equal to 180 degrees , type II tumors had more than 180 degrees and less than 270 degrees , and type III tumors had a maximum circumference of contact of 270 degrees of more. MR imaging accurately predicted the Shamblin group in 8 (100%) of 8 operated tumors. Tumor size and Shamblin group did not have a uniformly predictable relation. CONCLUSIONS: Shamblin group can be predicted preoperatively on MR imaging, and the maximum degree of circumferential contact of the CBT with the ICA on axial images is the criterion to predict the Shamblin group. PMID- 18417603 TI - Brain parenchymal signal abnormalities associated with developmental venous anomalies: detailed MR imaging assessment. AB - BACKGROUND AND PURPOSE: The occurrence of brain parenchymal signal-intensity changes within the drainage territory of developmental venous anomalies (DVAs) in the absence of cavernous malformations (CMs) has been incompletely assessed. This study was performed to evaluate the prevalence of brain parenchymal signal intensity abnormalities subjacent to DVA, correlating with DVA morphology and location. MATERIALS AND METHODS: One hundred sixty-four patients with brain MR imaging with contrast studies performed from July 2005 through June 2006 formed the study group. The examinations were reviewed and data were collected regarding the following: location, depth, size of draining vein, associated increased signal intensity on fluid-attenuated inversion recovery and T2-weighted images, associated CMs, and associated signal intensity on gradient recalled-echo sequences. RESULTS: Of the 175 DVAs identified, 28 had associated signal intensity abnormalities in the drainage territory. Seven of 28 DVAs with signal intensity abnormalities were excluded because of significant adjacent white matter signal-intensity changes related to other pathology overlapping the drainage territory. Of the remaining DVAs imaged in this study, 21/168 (12.5%) had subjacent signal-intensity abnormalities. An adjusted prevalence rate of 9/115 (7.8%) was obtained by excluding patients with white matter disease more than minimal in degree. Periventricular location and older age were associated with DVA signal-intensity abnormality. CONCLUSION: Signal-intensity abnormalities detectable by standard clinical MR images were identified in association with 12.5% of consecutively identified DVAs. Excluding patients with significant underlying white matter disease, we adjusted the prevalence to 7.8%. The etiology of the signal-intensity changes is unclear but may be related to edema, gliosis, or leukoaraiosis secondary to altered hemodynamics in the drainage area. PMID- 18417604 TI - The oculomotor cistern: anatomy and high-resolution imaging. AB - BACKGROUND AND PURPOSE: The oculomotor cistern (OMC) is a small CSF-filled dural cuff that invaginates into the cavernous sinus, surrounding the third cranial nerve (CNIII). It is used by neurosurgeons to mobilize CNIII during cavernous sinus surgery. In this article, we present the OMC imaging spectrum as delineated on 1.5T and 3T MR images and demonstrate its involvement in cavernous sinus pathology. MATERIALS AND METHODS: We examined 78 high-resolution screening MR images of the internal auditory canals (IAC) obtained for sensorineural hearing loss. Cistern length and diameter were measured. Fifty randomly selected whole brain MR images were evaluated to determine how often the OMC can be visualized on routine scans. Three volunteers underwent dedicated noncontrast high resolution MR imaging for optimal OMC visualization. RESULTS: One or both OMCs were visualized on 75% of IAC screening studies. The right cistern length averaged 4.2 +/- 3.2 mm; the opening diameter (the porus) averaged 2.2 +/- 0.8 mm. The maximal length observed was 13.1 mm. The left cistern length averaged 3.0 +/- 1.7 mm; the porus diameter averaged 2.1 +/-1.0 mm, with a maximal length of 5.9 mm. The OMC was visualized on 64% of routine axial T2-weighted brain scans. CONCLUSION: The OMC is an important neuroradiologic and surgical landmark, which can be routinely identified on dedicated thin-section high-resolution MR images. It can also be identified on nearly two thirds of standard whole-brain MR images. PMID- 18417605 TI - Coils in a nutshell: a review of coil physical properties. AB - Since its inception, endovascular coil technology has grown substantially as multiple manufacturers entered the market with an ever-increasing number of new products. Practitioners are now inundated with a choice of coils that vary on the basis of factors such as size, composition, stiffness, and detachment mechanism. The seasoned interventionalist had the benefit of evolving with this technology and, therefore, is likely to understand many of the practical nuances of coil development; for more junior practitioners, who did not experience the ongoing changes in technology, this review will provide a basic framework for the fundamentals of coil design. PMID- 18417606 TI - Significance and targeting of small, central clefts in severe fractures treated with vertebroplasty. AB - We report a small cohort of patients with severe osteoporotic fractures treated with vertebroplasty. We note a high prevalence of small, central, intraosseous clefts in these severe fractures. Rather than filling the small amount of residual bone marrow around the periphery of these severe fractures, as suggested by previous authors, we suggest central needle placement to fill these central clefts. PMID- 18417607 TI - Diffusion tensor MR imaging of the neurologically intact human spinal cord. AB - BACKGROUND AND PURPOSE: The aim of this study was to characterize the diffusion properties of the entire human spinal cord in vivo. These data are essential for comparisons to pathologic conditions as well as for comparisons of different pulse sequence design parameters aimed to reduce scan time and more accurately determine diffusion coefficients. MATERIALS AND METHODS: A total of 13 neurologically intact subjects were enrolled in this study. A single-shot, twice refocused, spin-echo, diffusion-weighted, echo-planar imaging (EPI) pulse sequence was used to obtain axial images throughout the entire spinal cord (C1 L1) in 45 minutes. RESULTS: Diffusion images indicated slight geometric distortions; however, gray and white matter contrast was observed. All measurements varied across the length of the cord. Whole cord diffusion coefficients averaged 0.5-1.3 x 10(-3) mm(2)/s depending on orientation, mean diffusivity (MD) averaged 0.83 +/- 0.06 x 10(-3) mm(2)/s, fractional anisotropy (FA) averaged 0.49 +/- 0.05, and volume ratio (VR) averaged 0.73 +/- 0.05. CONCLUSION: This study provided normative diffusion values for the entire spinal cord for use in comparisons with pathologic conditions as well as improvements in pulse sequence design. PMID- 18417608 TI - Diffuse pachymeningeal hyperintensity and subdural effusion/hematoma detected by fluid-attenuated inversion recovery MR imaging in patients with spontaneous intracranial hypotension. AB - BACKGROUND AND PURPOSE: Fluid-attenuated inversion recovery (FLAIR) MR imaging has advantages to detect meningeal lesions. FLAIR MR imaging was used to detect pachymeningeal thickening and thin bilateral subdural effusion/hematomas in patients with spontaneous intracranial hypotension (SIH). MATERIALS AND METHODS: Eight patients were treated under clinical diagnoses of SIH. Chronologic MR imaging studies, including the FLAIR sequence, were retrospectively reviewed. RESULTS: Initial MR imaging showed diffuse pachymeningeal thickening as isointense in 6 cases, hypoisointense in 1 case, and isohyperintense in 1 case on the T1-weighted MR images, and hyperintense in all cases on both T2-weighted and FLAIR MR images. Dural (pachymeningeal) hyperintensity on FLAIR MR imaging had the highest contrast to CSF, and was observed as linear in all patients, usually located in the supratentorial convexity and also parallel to the falx, the dura of the posterior fossa convexity, and the tentorium, and improved after treatment. These characteristics of diffuse pachymeningeal hyperintensity on FLAIR MR imaging were similar to diffuse pachymeningeal enhancement (DPME) on T1 weighted imaging with gadolinium. Initial FLAIR imaging clearly showed subdural effusion/hematomas in 6 of 8 patients. The thickness of subdural effusion/hematomas sometimes increased transiently after successful treatment and resolution of clinical symptoms. CONCLUSION: Diffuse pachymeningeal hyperintensity on FLAIR MR imaging is a similar sign to DPME for the diagnosis of SIH but does not require injection of contrast medium. FLAIR is useful sequence for the detection of subdural effusion/hematomas in patients with SIH. PMID- 18417609 TI - Identification of a novel protein MICS1 that is involved in maintenance of mitochondrial morphology and apoptotic release of cytochrome c. AB - Mitochondrial morphology dynamically changes in a balance of membrane fusion and fission in response to the environment, cell cycle, and apoptotic stimuli. Here, we report that a novel mitochondrial protein, MICS1, is involved in mitochondrial morphology in specific cristae structures and the apoptotic release of cytochrome c from the mitochondria. MICS1 is an inner membrane protein with a cleavable presequence and multiple transmembrane segments and belongs to the Bi-1 super family. MICS1 down-regulation causes mitochondrial fragmentation and cristae disorganization and stimulates the release of proapoptotic proteins. Expression of the anti-apoptotic protein Bcl-XL does not prevent morphological changes of mitochondria caused by MICS1 down-regulation, indicating that MICS1 plays a role in maintaining mitochondrial morphology separately from the function in apoptotic pathways. MICS1 overproduction induces mitochondrial aggregation and partially inhibits cytochrome c release during apoptosis, regardless of the occurrence of Bax targeting. MICS1 is cross-linked to cytochrome c without disrupting membrane integrity. Thus, MICS1 facilitates the tight association of cytochrome c with the inner membrane. Furthermore, under low-serum condition, the delay in apoptotic release of cytochrome c correlates with MICS1 up-regulation without significant changes in mitochondrial morphology, suggesting that MICS1 individually functions in mitochondrial morphology and cytochrome c release. PMID- 18417610 TI - Analysis of the yeast kinome reveals a network of regulated protein localization during filamentous growth. AB - The subcellular distribution of kinases and other signaling proteins is regulated in response to cellular cues; however, the extent of this regulation has not been investigated for any gene set in any organism. Here, we present a systematic analysis of protein kinases in the budding yeast, screening for differential localization during filamentous growth. Filamentous growth is an important stress response involving mitogen-activated protein kinase and cAMP-dependent protein kinase signaling modules, wherein yeast cells form interconnected and elongated chains. Because standard strains of yeast are nonfilamentous, we constructed a unique set of 125 kinase-yellow fluorescent protein chimeras in the filamentous Sigma1278b strain for this study. In total, we identified six cytoplasmic kinases (Bcy1p, Fus3p, Ksp1p, Kss1p, Sks1p, and Tpk2p) that localize predominantly to the nucleus during filamentous growth. These kinases form part of an interdependent, localization-based regulatory network: deletion of each individual kinase, or loss of kinase activity, disrupts the nuclear translocation of at least two other kinases. In particular, this study highlights a previously unknown function for the kinase Ksp1p, indicating the essentiality of its nuclear translocation during yeast filamentous growth. Thus, the localization of Ksp1p and the other kinases identified here is tightly controlled during filamentous growth, representing an overlooked regulatory component of this stress response. PMID- 18417611 TI - Gcn4 is required for the response to peroxide stress in the yeast Saccharomyces cerevisiae. AB - An oxidative stress occurs when reactive oxygen species overwhelm the cellular antioxidant defenses. We have examined the regulation of protein synthesis in Saccharomyces cerevisiae in response to oxidative stress induced by exposure to hydroperoxides (hydrogen peroxide, and cumene hydroperoxide), a thiol oxidant (diamide), and a heavy metal (cadmium). Examination of translational activity indicates that these oxidants inhibit translation at the initiation and postinitiation phases. Inhibition of translation initiation in response to hydroperoxides is entirely dependent on phosphorylation of the alpha subunit of eukaryotic initiation factor (eIF)2 by the Gcn2 kinase. Activation of Gcn2 is mediated by uncharged tRNA because mutation of its HisRS domain abolishes regulation in response to hydroperoxides. Furthermore, Gcn4 is translationally up regulated in response to H(2)O(2), and it is required for hydroperoxide resistance. We used transcriptional profiling to identify a wide range of genes that mediate this response as part of the Gcn4-dependent H(2)O(2)-regulon. In contrast to hydroperoxides, regulation of translation initiation in response to cadmium and diamide depends on both Gcn2 and the eIF4E binding protein Eap1. Thus, the response to oxidative stress is mediated by oxidant-specific regulation of translation initiation, and we suggest that this is an important mechanism underlying the ability of cells to adapt to different oxidants. PMID- 18417612 TI - The Rho GDI Rdi1 regulates Rho GTPases by distinct mechanisms. AB - The small guanosine triphosphate (GTP)-binding proteins of the Rho family are implicated in various cell functions, including establishment and maintenance of cell polarity. Activity of Rho guanosine triphosphatases (GTPases) is not only regulated by guanine nucleotide exchange factors and GTPase-activating proteins but also by guanine nucleotide dissociation inhibitors (GDIs). These proteins have the ability to extract Rho proteins from membranes and keep them in an inactive cytosolic complex. Here, we show that Rdi1, the sole Rho GDI of the yeast Saccharomyces cerevisiae, contributes to pseudohyphal growth and mitotic exit. Rdi1 interacts only with Cdc42, Rho1, and Rho4, and it regulates these Rho GTPases by distinct mechanisms. Binding between Rdi1 and Cdc42 as well as Rho1 is modulated by the Cdc42 effector and p21-activated kinase Cla4. After membrane extraction mediated by Rdi1, Rho4 is degraded by a novel mechanism, which includes the glycogen synthase kinase 3beta homologue Ygk3, vacuolar proteases, and the proteasome. Together, these results indicate that Rdi1 uses distinct modes of regulation for different Rho GTPases. PMID- 18417613 TI - Redundant roles of BIG2 and BIG1, guanine-nucleotide exchange factors for ADP ribosylation factors in membrane traffic between the trans-Golgi network and endosomes. AB - BIG2 and BIG1 are closely related guanine-nucleotide exchange factors (GEFs) for ADP-ribosylation factors (ARFs) and are involved in the regulation of membrane traffic through activating ARFs and recruiting coat protein complexes, such as the COPI complex and the AP-1 clathrin adaptor complex. Although both ARF-GEFs are associated mainly with the trans-Golgi network (TGN) and BIG2 is also associated with recycling endosomes, it is unclear whether BIG2 and BIG1 share some roles in membrane traffic. We here show that knockdown of both BIG2 and BIG1 by RNAi causes mislocalization of a subset of proteins associated with the TGN and recycling endosomes and blocks retrograde transport of furin from late endosomes to the TGN. Similar mislocalization and protein transport block, including furin, were observed in cells depleted of AP-1. Taken together with previous reports, these observations indicate that BIG2 and BIG1 play redundant roles in trafficking between the TGN and endosomes that involves the AP-1 complex. PMID- 18417614 TI - Glypican-1 regulates anaphase promoting complex/cyclosome substrates and cell cycle progression in endothelial cells. AB - Glypican-1 (GPC1), a member of the mammalian glypican family of heparan sulfate proteoglycans, is highly expressed in glioma blood vessel endothelial cells (ECs). In this study, we investigated the role of GPC1 in EC replication by manipulating GPC1 expression in cultured mouse brain ECs. Moderate GPC1 overexpression stimulates EC growth, but proliferation is significantly suppressed when GPC1 expression is either knocked down or the molecule is highly overexpressed. Flow cytometric and biochemical analyses show that high or low expression of GPC1 causes cell cycle arrest at mitosis or the G2 phase of the cell cycle, accompanied by endoreduplication and consequently polyploidization. We further show that GPC1 inhibits the anaphase-promoting complex/cyclosome (APC/C)-mediated degradation of mitotic cyclins and securin. High levels of GPC1 induce metaphase arrest and centrosome overproduction, alterations that are mimicked by overexpression of cyclin B1 and cyclin A, respectively. These observations suggest that GPC1 regulates EC cell cycle progression at least partially by modulating APC/C-mediated degradation of mitotic cyclins and securin. PMID- 18417615 TI - The subcellular distribution of calnexin is mediated by PACS-2. AB - Calnexin is an endoplasmic reticulum (ER) lectin that mediates protein folding on the rough ER. Calnexin also interacts with ER calcium pumps that localize to the mitochondria-associated membrane (MAM). Depending on ER homeostasis, varying amounts of calnexin target to the plasma membrane. However, no regulated sorting mechanism is so far known for calnexin. Our results now describe how the interaction of calnexin with the cytosolic sorting protein PACS-2 distributes calnexin between the rough ER, the MAM, and the plasma membrane. Under control conditions, more than 80% of calnexin localizes to the ER, with the majority on the MAM. PACS-2 knockdown disrupts the calnexin distribution within the ER and increases its levels on the cell surface. Phosphorylation by protein kinase CK2 of two calnexin cytosolic serines (Ser554/564) reduces calnexin binding to PACS 2. Consistent with this, a Ser554/564 Asp phosphomimic mutation partially reproduces PACS-2 knockdown by increasing the calnexin signal on the cell surface and reducing it on the MAM. PACS-2 knockdown does not reduce retention of other ER markers. Therefore, our results suggest that the phosphorylation state of the calnexin cytosolic domain and its interaction with PACS-2 sort this chaperone between domains of the ER and the plasma membrane. PMID- 18417616 TI - The endocytic adaptor protein ARH associates with motor and centrosomal proteins and is involved in centrosome assembly and cytokinesis. AB - Numerous proteins involved in endocytosis at the plasma membrane have been shown to be present at novel intracellular locations and to have previously unrecognized functions. ARH (autosomal recessive hypercholesterolemia) is an endocytic clathrin-associated adaptor protein that sorts members of the LDL receptor superfamily (LDLR, megalin, LRP). We report here that ARH also associates with centrosomes in several cell types. ARH interacts with centrosomal (gamma-tubulin and GPC2 and GPC3) and motor (dynein heavy and intermediate chains) proteins. ARH cofractionates with gamma-tubulin on isolated centrosomes, and gamma-tubulin and ARH interact on isolated membrane vesicles. During mitosis, ARH sequentially localizes to the nuclear membrane, kinetochores, spindle poles and the midbody. Arh(-/-) embryonic fibroblasts (MEFs) show smaller or absent centrosomes suggesting ARH plays a role in centrosome assembly. Rat-1 fibroblasts depleted of ARH by siRNA and Arh(-/-) MEFs exhibit a slower rate of growth and prolonged cytokinesis. Taken together the data suggest that the defects in centrosome assembly in ARH depleted cells may give rise to cell cycle and mitotic/cytokinesis defects. We propose that ARH participates in centrosomal and mitotic dynamics by interacting with centrosomal proteins. Whether the centrosomal and mitotic functions of ARH are related to its endocytic role remains to be established. PMID- 18417617 TI - Developmental origin of smooth muscle cells in the descending aorta in mice. AB - Aortic smooth muscle cells (SMCs) have been proposed to derive from lateral plate mesoderm. It has further been suggested that induction of SMC differentiation is confined to the ventral side of the aorta, and that SMCs later migrate to the dorsal side. In this study, we investigate the origin of SMCs in the descending aorta using recombination-based lineage tracing in mice. Hoxb6-cre transgenic mice were crossed with Rosa 26 reporter mice to track cells of lateral plate mesoderm origin. The contribution of lateral plate mesoderm to SMCs in the descending aorta was determined at different stages of development. SMC differentiation was induced in lateral plate mesoderm-derived cells on the ventral side of the aorta at embryonic day (E) 9.0-9.5, as indicated by expression of the SMC-specific reporter gene SM22alpha-lacZ. There was, however, no migration of SMCs from the ventral to the dorsal side of the vessel. Moreover, the lateral plate mesoderm-derived cells in the ventral wall of the aorta were replaced by somite-derived cells at E10.5, as indicated by reporter gene expression in Meox1-cre/Rosa 26 double transgenic mice. Examination of reporter gene expression in adult aortas from Hoxb6-cre/Rosa 26 and Meox1-cre/Rosa 26 double transgenic mice suggested that all SMCs in the adult descending aorta derive from the somites, whereas no contribution was recorded from lateral plate mesoderm. PMID- 18417618 TI - Zic2 promotes axonal divergence at the optic chiasm midline by EphB1-dependent and -independent mechanisms. AB - Axons of retinal ganglion cells (RGCs) make a divergent choice at the optic chiasm to cross or avoid the midline in order to project to ipsilateral and contralateral targets, thereby establishing the binocular visual pathway. The zinc-finger transcription factor Zic2 and a member of the Eph family of receptor tyrosine kinases, EphB1, are both essential for proper development of the ipsilateral projection at the mammalian optic chiasm midline. Here, we demonstrate in mouse by functional experiments in vivo that Zic2 is not only required but is also sufficient to change the trajectory of RGC axons from crossed to uncrossed. In addition, our results reveal that this transcription factor regulates the expression of EphB1 in RGCs and also suggest the existence of an additional EphB1-independent pathway controlled by Zic2 that contributes to retinal axon divergence at the midline. PMID- 18417619 TI - SoxB1 transcription factors and Notch signaling use distinct mechanisms to regulate proneural gene function and neural progenitor differentiation. AB - The preservation of a pool of neural precursors is a prerequisite for proper establishment and maintenance of a functional central nervous system (CNS). Both Notch signaling and SoxB1 transcription factors have been ascribed key roles during this process, but whether these factors use common or distinct mechanisms to control progenitor maintenance is unsettled. Here, we report that the capacity of Notch to maintain neural cells in an undifferentiated state requires the activity of SoxB1 proteins, whereas the mechanism by which SoxB1 block neurogenesis is independent of Notch signaling. A common feature of Notch signaling and SoxB1 proteins is their ability to inhibit the activity of proneural bHLH proteins. Notch represses the transcription of proneural bHLH genes, while SoxB1 proteins block their neurogenic capacity. Moreover, E-proteins act as functional partners of proneural proteins and the suppression of E-protein expression is an important mechanism by which Notch counteracts neurogenesis. Interestingly, in contrast to the Hes-dependent repression of proneural genes, suppression of E-protein occurs in a Hes-independent fashion. Together, these data reveal that Notch signaling and SoxB1 transcription factors use distinct regulatory mechanisms to control proneural protein function and to preserve neural cells as undifferentiated precursors. PMID- 18417620 TI - Regulation of TGF-(beta) signalling by N-acetylgalactosaminyltransferase-like 1. AB - The TGF-beta superfamily of secreted signalling molecules plays a pivotal role in the regulation of early embryogenesis, organogenesis and adult tissue homeostasis. Here we report the identification of Xenopus N acetylgalactosaminyltransferase-like 1 (xGalntl-1) as a novel important regulator of TGF-beta signalling. N-acetylgalactosaminyltransferases mediate the first step of mucin-type glycosylation, adding N-acetylgalactose to serine or threonine side chains. xGalntl-1 is expressed in the anterior mesoderm and neural crest territory at neurula stage, and in the anterior neural crest, notochord and the mediolateral spinal cord at tailbud stage. Inhibition of endogenous xGalntl-1 protein synthesis, using specific morpholino oligomers, interfered with the formation of anterior neural crest, anterior notochord and the spinal cord. Xenopus and mammalian Galntl-1 inhibited Activin as well as BMP signalling in the early Xenopus embryo and in human HEK 293T cells. Gain- and loss-of-function experiments showed that xGalntl-1 interferes with the activity of the common TGF beta type II receptor ActR-IIB in vivo. In addition, our biochemical data demonstrated that xGalntl-1 specifically interferes with the binding of ActR-IIB to Activin- and BMP-specific type I receptors. This inhibitory activity of xGalntl-1 was dependent on mucin-type glycosylation, as it was sensitive to the chemical inhibitor benzyl-GalNAc. These studies reveal an important role of a N acetylgalactosaminyltransferase in the regulation of TGF-beta signalling. This novel regulatory mechanism is evolutionarily conserved and, thus, might provide a new paradigm for the regulation of TGF-beta signalling in vertebrates. PMID- 18417621 TI - Essential role for PDGF signaling in ophthalmic trigeminal placode induction. AB - Much of the peripheral nervous system of the head is derived from ectodermal thickenings, called placodes, that delaminate or invaginate to form cranial ganglia and sense organs. The trigeminal ganglion, which arises lateral to the midbrain, forms via interactions between the neural tube and adjacent ectoderm. This induction triggers expression of Pax3, ingression of placode cells and their differentiation into neurons. However, the molecular nature of the underlying signals remains unknown. Here, we investigate the role of PDGF signaling in ophthalmic trigeminal placode induction. By in situ hybridization, PDGF receptor beta is expressed in the cranial ectoderm at the time of trigeminal placode formation, with the ligand PDGFD expressed in the midbrain neural folds. Blocking PDGF signaling in vitro results in a dose-dependent abrogation of Pax3 expression in recombinants of quail ectoderm with chick neural tube that recapitulate placode induction. In ovo microinjection of PDGF inhibitor causes a similar loss of Pax3 as well as the later placodal marker, CD151, and failure of neuronal differentiation. Conversely, microinjection of exogenous PDGFD increases the number of Pax3+ cells in the trigeminal placode and neurons in the condensing ganglia. Our results provide the first evidence for a signaling pathway involved in ophthalmic (opV) trigeminal placode induction. PMID- 18417622 TI - CD41+ cmyb+ precursors colonize the zebrafish pronephros by a novel migration route to initiate adult hematopoiesis. AB - Development of the vertebrate blood lineages is complex, with multiple waves of hematopoietic precursors arising in different embryonic locations. Monopotent, or primitive, precursors first give rise to embryonic macrophages or erythrocytes. Multipotent, or definitive, precursors are subsequently generated to produce the adult hematopoietic lineages. In both the zebrafish and the mouse, the first definitive precursors are committed erythromyeloid progenitors (EMPs) that lack lymphoid differentiation potential. We have previously shown that zebrafish EMPs arise in the posterior blood island independently from hematopoietic stem cells (HSCs). In this report, we demonstrate that a fourth wave of hematopoietic precursors arises slightly later in the zebrafish aorta/gonad/mesonephros (AGM) equivalent. We have identified and prospectively isolated these cells by CD41 (itga2b) and cmyb expression. Unlike EMPs, CD41(+) AGM cells colonize the thymus to generate rag2(+) T lymphocyte precursors. Timelapse imaging and lineage tracing analyses demonstrate that AGM-derived precursors use a previously undescribed migration pathway along the pronephric tubules to initiate adult hematopoiesis in the developing kidney, the teleostean equivalent of mammalian bone marrow. Finally, we have analyzed the gene expression profiles of EMPs and AGM precursors to better understand the molecular cues that pattern the first definitive hematopoietic cells in the embryo. Together, these studies suggest that expression of CD41 and cmyb marks nascent HSCs in the zebrafish AGM, and provide the means to further dissect HSC generation and function in the early vertebrate embryo. PMID- 18417623 TI - Multiple maternal proteins coordinate to restrict the translation of C. elegans nanos-2 to primordial germ cells. AB - Although germ cell formation has been relatively well understood in worms and insects, how germ cell-specific developmental programs are initiated is not clear. In Caenorhabditis elegans, translational activation of maternal nos-2 mRNA is the earliest known molecular event specific to the germline founder cell P(4). Cis-elements in nos-2 3'UTR have been shown to mediate translational control; however, the trans-acting proteins are not known. Here, we provide evidence that four maternal RNA-binding proteins, OMA-1, OMA-2, MEX-3 and SPN-4, bind nos-2 3'UTR to suppress its translation, and POS-1, another maternal RNA-binding protein, relieves this suppression in P(4). The POS-1: SPN-4 ratio in P(4) increases significantly over its precursor, P(3); and POS-1 competes with SPN-4 for binding to nos-2 RNA in vitro. We propose temporal changes in the relative concentrations of POS-1 and SPN-4, through their effect on the translational status of maternal mRNAs such as nos-2, initiate germ cell-specific developmental programs in C. elegans. PMID- 18417624 TI - Upregulation of the T-type calcium current in small rat sensory neurons after chronic constrictive injury of the sciatic nerve. AB - Recent data indicate that peripheral T-type Ca2+ channels are instrumental in supporting acute pain transmission. However, the function of these channels in chronic pain processing is less clear. To address this issue, we studied the expression of T-type Ca2+ currents in small nociceptive dorsal root ganglion (DRG) cells from L4-5 spinal ganglia of adult rats with neuropathic pain due to chronic constrictive injury (CCI) of the sciatic nerve. In control rats, whole cell recordings revealed that T-type currents, measured in 10 mM Ba2+ as a charge carrier, were present in moderate density (20 +/- 2 pA/pF). In rats with CCI, T type current density (30 +/- 3 pA/pF) was significantly increased, but voltage- and time-dependent activation and inactivation kinetics were not significantly different from those in controls. CCI-induced neuropathy did not significantly change the pharmacological sensitivity of T-type current in these cells to nickel. Collectively, our results indicate that CCI-induced neuropathy significantly increases T-type current expression in small DRG neurons. Our finding that T-type currents are upregulated in a CCI model of peripheral neuropathy and earlier pharmacological and molecular studies suggest that T-type channels may be potentially useful therapeutic targets for the treatment of neuropathic pain associated with partial mechanical injury to the sciatic nerve. PMID- 18417625 TI - Corticofugal projection inhibits the auditory thalamus through the thalamic reticular nucleus. AB - Electrical stimulation of the auditory cortex (AC) causes both facilitatory and inhibitory effects on the medial geniculate body (MGB). The purpose of this study was to identify the corticofugal inhibitory pathway to the MGB. We assessed two potential circuits: 1) the cortico-colliculo-thalamic circuit and 2) cortico reticulo-thalamic one. We compared intracellular responses of MGB neurons to electrical stimulation of the AC following bilateral ablation of the inferior colliculi (IC) or thalamic reticular nucleus (TRN) in anesthetized guinea pigs. Cortical stimulation with intact TRN could cause strong inhibitory effects on the MGB neurons. The corticofugal inhibition remained effective after bilateral IC ablation, but it was minimized after the TRN was lesioned with kainic acid. Synchronized TRN neuronal activity and MGB inhibitory postsynaptic potentials (IPSPs) were observed with multiple recordings. The results suggest that corticofugal inhibition traverses the corticoreticulothalamic pathway, indicating that the colliculi-geniculate inhibitory pathway is probably only for feedforward inhibition. PMID- 18417626 TI - Inhibitory interactions among olfactory glomeruli do not necessarily reflect spatial proximity. AB - Inhibitory interactions shape the activity of output neurons in primary olfactory centers and promote contrast enhancement of odor representations. Patterns of interglomerular connectivity, however, are largely unknown. To test whether the proximity of glomeruli to one another is correlated with interglomerular inhibitory interactions, we used intracellular recording and staining methods to record the responses of projection (output) neurons (PNs) associated with glomeruli of known olfactory tuning in the primary olfactory center of the moth Manduca sexta. We focused on Toroid I, a glomerulus in the male-specific macroglomerular complex (MGC) specialized to one of the two key components of the conspecific females' sex pheromone, and the adjacent, sexually isomorphic glomerulus 35, which is highly sensitive to Z-3-hexenyl acetate (Z3-6:OAc). We used the two odorants to activate these reference glomeruli and tested the effects of olfactory activation in other glomeruli. We found that Toroid-I PNs were not inhibited by input to G35, whereas G35 PNs were inhibited by input to Toroid-I PNs. We also recorded the responses of PNs arborizing in other sexually isomorphic glomeruli to stimulation with the sex pheromone and Z3-6:OAc. We found that inhibitory responses were not related to proximity to the MGC and G35: both distant and adjacent PNs were inhibited by stimulation with the sex pheromone, some others were affected by only one odorant, and yet others by neither. Similar results were obtained in female PNs recorded in proximity to female-specific glomeruli. Our findings indicate that inhibitory interactions among glomeruli are widespread and independent of their spatial proximity. PMID- 18417627 TI - Effects of anoxia and aglycemia on cytosolic calcium regulation in rat sensory neurons. AB - Nociceptive neurons play an important role in ischemia by sensing and transmitting information to the CNS and by secreting peptides and nitric oxide, which can have local effects. While these responses are probably primarily mediated by acid sensing channels, other events occurring in ischemia may also influence neuron function. In this study, we have investigated the effects of anoxia and anoxic aglycemia on Ca2+ regulation in sensory neurons from rat dorsal root ganglia. Anoxia increased [Ca2+]i by evoking Ca2+ release from two distinct internal stores one sensitive to carbonyl cyanide p-(trifluoromethoxy) phenylhydrazone (FCCP) and one sensitive to caffeine, cyclopiazonic acid (CPA), and ryanodine [assumed to be the endoplasmic reticulum (ER)]. Anoxia also promoted progressive decline in ER Ca2+ content. Despite partially depolarizing mitochondria, anoxia had relatively little effect on mitochondrial Ca2+ uptake when neurons were depolarized but substantially delayed mitochondrial Ca2+ release and subsequent Ca2+ clearance from the cytosol on repolarization. Anoxia also reduced both sarco(endo)plasmic reticulum Ca2+-ATPase (SERCA) activity and Ca2+ extrusion [probably via plasma membrane Ca2+-ATPase (PMCA)]. Thus anoxia has multiple effects on [Ca2+]i homeostasis in sensory neurons involving internal stores, mitochondrial buffering, and Ca2+ pumps. Under conditions of anoxic aglycemia, there was a biphasic and more profound elevation of [Ca2+]i, which was associated with complete ER Ca2+ store emptying and progressive, and eventually complete, inhibition of Ca2+ clearance by PMCA and SERCA. These data clearly show that loss of oxygen, and exhaustion of glycolytic substrates, can profoundly affect many aspects of cell Ca2+ regulation, and this may play an important role in modulating neuronal responses to ischemia. PMID- 18417628 TI - Altered dendritic integration in hippocampal granule cells of spatial learning impaired aged rats. AB - Glutamatergic transmission at central synapses undergoes activity-dependent and developmental changes. In the hippocampal dentate gyrus, the non-N-methyl d aspartate (NMDA) receptor component of field excitatory postsynaptic potentials (fEPSPs) increases with age in Fischer-344 rats. This effect may not depend on the animal's activity or experience but could be part of the developmental process. Age-dependent differences in synaptic transmission at the perforant path granule cell synapse may be caused by changes in non-NMDA and NMDA receptor mediated currents. To test this hypothesis, we compared whole cell excitatory postsynaptic currents (EPSCs) in dentate granule cells evoked by perforant path stimulation in young (3-4 mo) and aged (22-27 mo) Fischer-344 rats using a Cs+ based intracellular solution. Aged animals as a group showed spatial learning and memory deficits in the Morris water maze. Using whole cell recordings, slope conductances of both non-NMDA and NMDA EPSCs at holding potentials -10 to +50 mV were significantly reduced in aged animals and the non-NMDA/NMDA ratio in aged animals was found to be significantly smaller than in young animals. In contrast, we detected no differences in basic electrophysiological parameters, or absolute amplitudes of non-NMDA and NMDA EPSCs. Extracellular Cs+ increased the fEPSP in young slices to a greater degree than was found in the aged slices, while it increased population spikes to a greater degree in the aged rats. Our results not only provide evidence for reduced glutamatergic synaptic responses in Fischer-344 rats but also point to differential changes in Cs+-sensitive dendritic conductances, such as Ih or inwardly rectifying potassium currents, during aging. PMID- 18417629 TI - Synaptic commitment: developmentally regulated reciprocal changes in hippocampal granule cell NMDA and AMPA receptors over the lifespan. AB - Synaptic transmission in hippocampal field CA1 is largely N-methyl-d-aspartate receptor (NMDA(R)) dependent during the early postnatal period. It becomes increasingly mediated by alpha-amino-3-hydroxy-5-methylisoxazole-4-proprionate (AMPA) receptors until an adult ratio of AMPA to NMDA receptors is achieved. It is shown here that increases in the AMPA receptor (AMPA(R))-mediated field potential response continue over the life span of the F-344 rat at the perforant path-granule cell synapse in the dentate gyrus. In contrast, the NMDA(R) dependent component of the response decreases with age between 1 and 27 mo, leading to an increase of AMPA(R)/NMDA(R) ratio with age. One possible explanation of this age difference is that the AMPA(R)/NMDA(R) ratio can be modified by experience. To test the idea that the changed ratio is caused by the old rats' longer lives, an intensive 10-mo period of enrichment treatment was given to a group of animals, beginning at 3 mo of age. Compared with animals housed in standard cages, the enrichment treatment did not alter the glutamatergic response ratio measured with field potential recording methods. These data provide support for the conclusion that the observed change with age is developmentally regulated rather than experience dependent. Given the role of the NMDA(R) in synaptic plasticity, these changes suggest a progressive commitment of perforant path synapses to particular weights over the life span. One possible implication of this effect includes preservation of selected memories, ultimately at the expense of a reduced capacity to store new information. PMID- 18417631 TI - Maturation of intrinsic and synaptic properties of layer 2/3 pyramidal neurons in mouse auditory cortex. AB - We investigated the development of L2/3 pyramidal cell (PC) circuitry in juvenile mice from postnatal day 10 (P10) to P29. Using whole cell recordings in an in vitro thalamocortical slice preparation, we examined the connection architecture and intrinsic and synaptic properties of PCs. The excitatory connections between PCs were highly localized: the probability of connection between PCs declined with intersomatic distance from 0.18 to about 0.05 over 150 microm, but did not vary with age. However, the mean and variance of the intrinsic and synaptic properties of PCs changed dramatically between P10 and P29. The input resistance, membrane time constant, and resting membrane potential decreased, leading to reduced neural excitability in older animals. Likewise, there were age-dependent decreases in the amplitude and decay time of the excitatory postsynaptic potentials as well as short-term synaptic depression. Both the intrinsic and synaptic properties underwent a transitional period between P10 and P18 prior to reaching steady state at P19-P29. We show that these properties combine to produce age-related differential synaptic responses to low- and high-frequency synaptic input that may contribute to differences in auditory processing during development. PMID- 18417632 TI - Roll tilt psychophysics in rhesus monkeys during vestibular and visual stimulation. AB - How does the brain calculate the spatial orientation of the head relative to gravity? Psychophysical measurements are critical to investigate this question, but such measurements have been limited to humans. In non-human primates, behavioral measures have focused on vestibular-mediated eye movements, which do not reflect percepts of head orientation. We have therefore developed a method to measure tilt perception in monkeys, derived from the subjective visual vertical (SVV) task. Two rhesus monkeys were trained to align a light bar parallel to gravity and performed this task during roll tilts, centrifugation, and roll optokinetic stimulation. The monkeys accurately aligned the light bar with gravity during static roll tilts but also demonstrated small orientation dependent misperceptions of the tilt angle analogous to those measured in humans. When the gravito-inertial force (GIF) rotated dynamically in the roll plane, SVV responses remained closely aligned with the GIF during roll tilt of the head (coplanar canal rotational cues present), lagged slightly behind the GIF during variable-radius centrifugation (no canal cues present), and shifted gradually during fixed-radius centrifugation (orthogonal yaw canal cues present). SVV responses also deviated away from the earth-vertical during roll optokinetic stimulation. These results demonstrate that rotational cues derived from the semicircular canals and visual system have prominent effects on psychophysical measurements of roll tilt in rhesus monkeys and therefore suggest that a central synthesis of graviceptive and rotational cues contributes to percepts of head orientation relative to gravity in non-human primates. PMID- 18417630 TI - Delayed orexin signaling consolidates wakefulness and sleep: physiology and modeling. AB - Orexin-producing neurons are clearly essential for the regulation of wakefulness and sleep because loss of these cells produces narcolepsy. However, little is understood about how these neurons dynamically interact with other wake- and sleep-regulatory nuclei to control behavioral states. Using survival analysis of wake bouts in wild-type and orexin knockout mice, we found that orexins are necessary for the maintenance of long bouts of wakefulness, but orexin deficiency has little impact on wake bouts <1 min. Since orexin neurons often begin firing several seconds before the onset of waking, this suggests a surprisingly delayed onset (>1 min) of functional effects. This delay has important implications for understanding the control of wakefulness and sleep because increasing evidence suggests that different mechanisms are involved in the production of brief and sustained wake bouts. We incorporated these findings into a mathematical model of the mouse sleep/wake network. Orexins excite monoaminergic neurons and we hypothesize that orexins increase the monoaminergic inhibition of sleep-promoting neurons in the ventrolateral preoptic nucleus. We modeled orexin effects as a time-dependent increase in the strength of inhibition from wake- to sleep promoting populations and the resulting simulated behavior accurately reflects the fragmented sleep/wake behavior of narcolepsy and leads to several predictions. By integrating neurophysiology of the sleep/wake network with emergent properties of behavioral data, this model provides a novel framework for investigating network dynamics and mechanisms associated with normal and pathologic sleep/wake behavior. PMID- 18417633 TI - Adaptation of cutaneous stumble correction when tripping is part of the locomotor environment. AB - We recently showed that cutaneous reflexes evoked by stimulating the superficial peroneal (SP; innervates foot dorsum) nerve are modulated according to the level of postural threat. Context-related modulation was observed mainly in contralateral (c) responses but not in the ipsilateral responses. This lack of effect on ipsilateral (i) cutaneous reflexes might have been caused by the general nature of the whole body perturbation. We therefore hypothesized that context-relevant mechanical perturbations applied to the dorsum of the foot by an instrumented rod at early swing during walking would produce differences in ipsilateral cutaneous reflex amplitudes, consistent with the functional relevance of the SP nerve in stumble correction responses. Subjects walked on a motorized treadmill under four conditions: 1) normal, 2) normal with mechanical perturbations at the foot dorsum, 3) arms crossed, and 4) arms crossed with mechanical perturbations at the foot dorsum. Electrical stimulation of the SP nerve was delivered at five phases of the step cycle, and cutaneous reflexes were compared between all conditions for each phase of the step cycle. Reflex responses were generally found to be modulated in amplitude during walking conditions in which mechanical perturbations were delivered, particularly in ipsilateral tibialis anterior (iTA), which showed a marked reduction in inhibition. The results indicated cutaneous reflexes in iTA and contralateral medial gastrocnemius (cMG) were influenced by the threat of a trip, induced by applying mechanical perturbations to the foot dorsum during walking. This task related gating of cutaneous reflexes was not generalized to all muscles, thus suggesting a functional role in the maintenance of stability during locomotion. PMID- 18417634 TI - Nasal solitary chemoreceptor cell responses to bitter and trigeminal stimulants in vitro. AB - Nasal trigeminal chemosensitivity in mice and rats is mediated in part by epithelial solitary chemoreceptor (chemosensory) cells (SCCs), but the exact role of these cells in chemoreception is unclear. Histological evidence suggests that SCCs express elements of the bitter taste transduction pathway including T2R (bitter taste) receptors, the G protein alpha-gustducin, PLCbeta2, and TRPM5, leading to speculation that SCCs are the receptor cells that mediate trigeminal nerve responses to bitter taste receptor ligands. To test this hypothesis, we used calcium imaging to determine whether SCCs respond to classic bitter-tasting or trigeminal stimulants. SCCs from the anterior nasal cavity were isolated from transgenic mice in which green fluorescent protein (GFP) expression was driven by either TRPM5 or gustducin. Isolated cells were exposed to a variety of test stimuli to determine which substances caused an increase in intracellular Ca2+ ([Ca2+]i). GFP-positive cells respond with increased [Ca2+]i to the bitter receptor ligand denatonium and this response is blocked by the PLC inhibitor U73122. In addition, GFP+ cells respond to the neuromodulators adenosine 5' triphosphate and acetylcholine but only very rarely to other bitter-tasting or trigeminal stimuli. Our results demonstrate that TRPM5- and gustducin-expressing nasal SCCs respond to the T2R agonist denatonium via a PLC-coupled transduction cascade typical of T2Rs in the taste system. PMID- 18417635 TI - Differential cholinergic modulation of Ca2+ transients evoked by backpropagating action potentials in apical and basal dendrites of cortical pyramidal neurons. AB - The effect of the cholinergic agonist carbachol (CCh) on backpropagating action potential (bAP)-evoked Ca2+ transients in distal apical and basal dendrites of layer 2/3 pyramidal neurons in the primary visual cortex of rats was studied using whole cell recordings and confocal Ca2+ imaging. In the presence of CCh (20 microM), initial bAP-evoked Ca2+ transients were followed by large propagating secondary Ca2+ transients that were restricted to proximal apical dendrites < or =40 microm from the soma. In middle apical dendrites (41-100 microm from the soma), Ca2+ transients evoked by AP bursts at 20 Hz, but not by single APs, were increased by CCh without secondary transients. CCh failed to increase the bAP evoked Ca2+ transients in distal apical dendrites (101-270 microm from the soma). In contrast, in basal dendrites, CCh increased Ca2+ transients evoked by AP bursts, but not by single APs, and these transients were relatively constant over the entire length of the dendrites. CCh further increased the enhanced bAP-evoked Ca2+ transients in the presence of 4-aminopyridine (200 microM), an A-type K+ channel blocker, in basal and apical dendrites, except in distal apical dendrites. CCh increased large Ca2+ transients evoked by high-frequency AP bursts in basal dendrites, but not in distal apical dendrites. CCh-induced increase in Ca2+ transients was mediated by InsP3-dependent Ca2+-induced Ca2+-release. These results suggest that cholinergic stimulation differentially increases the bAP evoked increase in [Ca2+]i in apical and basal dendrites, which may modulate synaptic activities in a location-dependent manner. PMID- 18417636 TI - Three-dimensional gas exchange pathways in pome fruit characterized by synchrotron x-ray computed tomography. AB - Our understanding of the gas exchange mechanisms in plant organs critically depends on insights in the three-dimensional (3-D) structural arrangement of cells and voids. Using synchrotron radiation x-ray tomography, we obtained for the first time high-contrast 3-D absorption images of in vivo fruit tissues of high moisture content at 1.4-microm resolution and 3-D phase contrast images of cell assemblies at a resolution as low as 0.7 microm, enabling visualization of individual cell morphology, cell walls, and entire void networks that were previously unknown. Intercellular spaces were always clear of water. The apple (Malus domestica) cortex contains considerably larger parenchyma cells and voids than pear (Pyrus communis) parenchyma. Voids in apple often are larger than the surrounding cells and some cells are not connected to void spaces. The main voids in apple stretch hundreds of micrometers but are disconnected. Voids in pear cortex tissue are always smaller than parenchyma cells, but each cell is surrounded by a tight and continuous network of voids, except near brachyssclereid groups. Vascular and dermal tissues were also measured. The visualized network architecture was consistent over different picking dates and shelf life. The differences in void fraction (5.1% for pear cortex and 23.0% for apple cortex) and in gas network architecture helps explain the ability of tissues to facilitate or impede gas exchange. Structural changes and anisotropy of tissues may eventually lead to physiological disorders. A combined tomography and internal gas analysis during growth are needed to make progress on the understanding of void formation in fruit. PMID- 18417637 TI - Overexpression of poplar cellulase accelerates growth and disturbs the closing movements of leaves in sengon. AB - In this study, poplar (Populus alba) cellulase (PaPopCel1) was overexpressed in a tropical Leguminosae tree, sengon (Paraserianthes falcataria), by the Agrobacterium tumefaciens method. PaPopCel1 overexpression increased the length and width of stems with larger leaves, which showed a moderately higher density of green color than leaves of the wild type. The pairs of leaves on the transgenic plants closed more slowly during sunset than those on the wild-type plants. When main veins from each genotype were excised and placed on a paper towel, however, the leaves of the transgenic plants closed more rapidly than those of the wild-type plant. Based on carbohydrate analyses of cell walls, the leaves of the transgenic plants contained less wall-bound xyloglucan than those of the wild-type plants. In situ xyloglucan endotransglucosylase activity showed that the incorporation of whole xyloglucan, potentially for wall tightening, occurred in the parenchyma cells (motor cells) of the petiolule pulvinus attached to the main vein, although the transgenic plant incorporated less whole xyloglucan than the wild-type plant. These observations support the hypothesis that the paracrystalline sites of cellulose microfibrils are attacked by poplar cellulase, which loosens xyloglucan intercalation, resulting in an irreversible wall modification. This process could be the reason why the overexpression of poplar cellulase both promotes plant growth and disturbs the biological clock of the plant by altering the closing movements of the leaves of the plant. PMID- 18417638 TI - A diurnal component to the variation in sieve tube amino acid content in wheat. AB - We have used high-sensitivity capillary electrophoresis coupled to a laser induced fluorescence detection method to quantify 16 amino acids in wheat (Triticum aestivum) sieve tube (ST) samples as small as 2 nL collected by severing the stylets of feeding aphids. The sensitivity of the method was sufficient to determine a quantitative amino acid profile of individual STs without the need to bulk samples to produce larger volumes for analysis. This allowed the observation of the full range of variation that exists in individual STs. Some of the total concentrations of amino acids recorded are higher than those reported previously. The results obtained show variation in the concentrations of phenylalanine (Phe), histidine/valine (His/Val), leucine/isoleucine (Leu/Ile), arginine, asparagine, glutamine, tyrosine (Tyr), and lysine (Lys) across the ST samples. These could not be explained by plant-to plant variation. Statistical analyses revealed five analytes (Tyr, Lys, Phe, His/Val, and Leu/Ile) that showed striking covariation in their concentrations across ST samples. A regression analysis revealed a significant relationship between the concentrations of Tyr, Lys, Phe, Leu/Ile, His/Val, asparagine, arginine, and proline and the time of collection of ST samples, with these amino acids increasing in concentration during the afternoon. This increase was confirmed to occur in individual STs by analyzing samples obtained from stylet bundles exuding for many hours. Finally, an apparent relationship between the exudation rate of ST sap and its total amino acid concentration was observed: samples containing higher total amino acid concentrations were observed to exude from the severed stylet bundles more slowly. PMID- 18417639 TI - Two GATA transcription factors are downstream effectors of floral homeotic gene action in Arabidopsis. AB - Floral organogenesis is dependent on the combinatorial action of MADS-box transcription factors, which in turn control the expression of suites of genes required for growth, patterning, and differentiation. In Arabidopsis (Arabidopsis thaliana), the specification of petal and stamen identity depends on the action of two MADS-box gene products, APETALA3 (AP3) and PISTILLATA (PI). In a screen for genes whose expression was altered in response to the induction of AP3 activity, we identified GNC (GATA, nitrate-inducible, carbon-metabolism-involved) as being negatively regulated by AP3 and PI. The GNC gene encodes a member of the Arabidopsis GATA transcription factor family and has been implicated in the regulation of chlorophyll biosynthesis as well as carbon and nitrogen metabolism. In addition, we found that the GNC paralog, GNL (GNC-like), is also negatively regulated by AP3 and PI. Using chromatin immunoprecipitation, we showed that promoter sequences of both GNC and GNL are bound by PI protein, suggesting a direct regulatory interaction. Analyses of single and double gnc and gnl mutants indicated that the two genes share redundant roles in promoting chlorophyll biosynthesis, suggesting that in repressing GNC and GNL, AP3/PI have roles in negatively regulating this biosynthetic pathway in flowers. In addition, coexpression analyses of genes regulated by AP3, PI, GNC, and GNL indicate a complex regulatory interplay between these transcription factors in regulating a variety of light and nutrient responsive genes. Together, these results provide new insights into the transcriptional cascades controlling the specification of floral organ identities. PMID- 18417640 TI - Serum 25-hydroxyvitamin D levels and the prevalence of peripheral arterial disease: results from NHANES 2001 to 2004. AB - OBJECTIVE: The purpose of this study was to determine the association between 25 hydroxyvitamin D (25(OH)D) levels and the prevalence of peripheral arterial disease (PAD) in the general United States population. METHODS AND RESULTS: We analyzed data from 4839 participants of the National Health and Nutrition Examination Survey 2001 to 2004 to evaluate the relationship between 25(OH)D and PAD (defined as an ankle-brachial index < 0.9). Across quartiles of 25(OH)D, from lowest to highest, the prevalence of PAD was 8.1%, 5.4%, 4.9%, and 3.7% (P trend < 0.001). After multivariable adjustment for demographics, comorbidities, physical activity level, and laboratory measures, the prevalence ratio of PAD for the lowest, compared to the highest, 25(OH)D quartile (< 17.8 and > or = 29.2 ng/mL, respectively) was 1.80 (95% confidence interval: 1.19, 2.74). For each 10 ng/mL lower 25(OH)D level, the multivariable-adjusted prevalence ratio of PAD was 1.35 (95% confidence interval: 1.15, 1.59). CONCLUSIONS: Low serum 25(OH)D levels are associated with a higher prevalence of PAD. Several mechanisms have been invoked in the literature to support a potential antiatherosclerotic activity of vitamin D. Prospective cohort and mechanistic studies should be designed to confirm this association. PMID- 18417641 TI - Peroxisome proliferator-activated receptor alpha activation regulates lipid metabolism in the feto-placental unit from diabetic rats. AB - Maternal diabetes promotes an overaccumulation of lipids in the feto-placental unit and impairs feto-placental development and growth. Here, we investigated the role played by the nuclear receptor peroxisome proliferator-activated receptor (PPAR)alpha in lipid metabolism in fetuses and placentas from control and neonatal streptozotocin-induced diabetic rats. Placentas and fetuses were studied on day 13.5 of gestation. The concentrations of PPARalpha (by Western blot) and its endogenous agonist leukotriene B(4) (LTB(4)) (by enzyme immunoassay) were analysed. Placental explants and fetuses were cultured with LTB(4) or clofibrate, and then lipid metabolism analysed (concentrations and synthesis from (14)C acetate of triglycerides, phospholipids, cholesterol and cholesteryl esters; release of glycerol and free fatty acids (FFAs)). We found that maternal diabetes led to increases in placental concentrations of triglycerides and cholesteryl esters, and fetal concentrations of phospholipids. PPARalpha agonists downregulated fetal and placental lipid concentrations in control and diabetic rats. The synthesis of lipids was reduced in the diabetic placenta but increased in fetuses from diabetic animals. PPARalpha agonists reduced the synthesis of lipids in control placenta and in the fetuses from control and diabetic rats. Glycerol and FFA release was enhanced in the diabetic placenta and in control placenta cultured with PPARalpha agonists. Maternal diabetes led to reductions in fetal and placental LTB(4) concentrations and to increases in placental PPARalpha concentrations. Overall, these data support a novel role of PPARalpha as a regulator of lipid metabolism in the feto-placental unit, relevant in maternal diabetes where fetal and placental PPARalpha, LTB(4) and lipid concentrations are altered. PMID- 18417642 TI - Reappraisal of H2S/sulfide concentration in vertebrate blood and its potential significance in ischemic preconditioning and vascular signaling. AB - Hydrogen sulfide (H(2)S) is rapidly emerging as a biologically significant signaling molecule. Studies published before 2000 report low or undetectable H(2)S (usually as total sulfide) levels in blood or plasma, whereas recent work has reported sulfide concentrations between 10 and 300 microM, suggesting it acts as a circulating signal. In the first series of experiments, we used a recently developed polarographic sensor to measure the baseline level of endogenous H(2)S gas and turnover of exogenous H(2)S gas in real time in blood from numerous animals, including lamprey, trout, mouse, rat, pig, and cow. We found that, contrary to recent reports, H(2)S gas was essentially undetectable (<100 nM total sulfide) in all animals. Furthermore, exogenous sulfide was rapidly removed from blood, plasma, or 5% bovine serum albumin in vitro and from intact trout in vivo. To determine if blood H(2)S could transiently increase, we measured oxygen dependent H(2)S production by trout hearts in vitro and in vivo. H(2)S has been shown to mediate ischemic preconditioning (IPC) in mammals. IPC is present in trout and, unlike mammals, the trout myocardium obtains its oxygen from relatively hypoxic systemic venous blood. In vitro, myocardial H(2)S production was inversely related to Po(2), whereas we failed to detect H(2)S in ventral aortic blood from either normoxic or hypoxic fish in vivo. These results provide an autocrine or paracrine mechanism for myocardial coupling of hypoxia to H(2)S in IPC, i.e., oxygen sensing, but they fail to provide any evidence that H(2)S signaling is mediated by the circulation. PMID- 18417643 TI - Gender differences in trauma theory vs. practice: Comments on "Mechanism of estrogen-mediated intestinal protection following trauma-hemorrhage: p38 MAPK dependent upregulation of HO-1" by Hsu JT et al. PMID- 18417644 TI - Dynamics of sleep stage transitions in healthy humans and patients with chronic fatigue syndrome. AB - Physiological and/or pathological implications of the dynamics of sleep stage transitions have not, to date, been investigated. We report detailed duration and transition statistics between sleep stages in healthy subjects and in others with chronic fatigue syndrome (CFS); in addition, we also compare our data with previously published results for rats. Twenty-two healthy females and 22 female patients with CFS, characterized by complaints of unrefreshing sleep, underwent one night of polysomnographic recording. We find that duration of deep sleep (stages III and IV) follows a power-law probability distribution function; in contrast, stage II sleep durations follow a stretched exponential and stage I, and REM sleep durations follow an exponential function. These stage duration distributions show a gradually increasing departure from the exponential form with increasing depth of sleep toward a power-law type distribution for deep sleep, suggesting increasing complexity of regulation of deeper sleep stages. We also find a substantial number of REM to non-REM sleep transitions in humans, while this transition is reported to be virtually nonexistent in rats. The relative frequency of this REM to non-REM sleep transition is significantly lower in CFS patients than in controls, resulting in a significantly greater relative transition frequency of moving from both REM and stage I sleep to awake. Such an alteration in the transition pattern suggests that the normal continuation of sleep in light or REM sleep is disrupted in CFS. We conclude that dynamic transition analysis of sleep stages is useful for elucidating yet-to-be determined human sleep regulation mechanisms with pathophysiological implications. PMID- 18417645 TI - Effect of interval versus continuous training on cardiorespiratory and mitochondrial functions: relationship to aerobic performance improvements in sedentary subjects. AB - The goal of the study was to determine the effects of continuous (CT) vs. intermittent (IT) training yielding identical mechanical work and training duration on skeletal muscle and cardiorespiratory adaptations in sedentary subjects. Eleven subjects (6 men and 5 women, 45 +/- 3 years) were randomly assigned to either of the two 8-wk training programs in a cross-over design, separated by 12 wk of detraining. Maximal oxygen uptake (Vo2max) increased after both trainings (9% with CT vs. 15% with IT), whereas only IT was associated with faster Vo2 kinetics (tau: 68.0 +/- 1.6 vs. 54.9 +/- 0.7 s, P < 0.05) measured during a test to exhaustion (TTE) and with improvements in maximal cardiac output (Qmax, from 18.1 +/- 1.1 to 20.1 +/- 1.2 l/min; P < 0.01). Skeletal muscle mitochondrial oxidative capacities (Vmax) were only increased after IT (3.3 +/- 0.4 before and 4.5 +/- 0.6 micromol O2 x min(-1) x g dw(-1) after training; P < 0.05), whereas capillary density increased after both trainings, with a two-fold higher enhancement after CT (+21 +/- 1% for IT and +40 +/- 3% after CT, P < 0.05). The gain of Vmax was correlated with the gain of TTE and the gain of Vo2max with IT. The gain of Qmax was also correlated with the gain of VO2max. These results suggest that fluctuations of workload and oxygen uptake during training sessions, rather than exercise duration or global energy expenditure, are key factors in improving muscle oxidative capacities. In an integrative view, IT seems optimal in maximizing both peripheral muscle and central cardiorespiratory adaptations, permitting significant functional improvement. These data support the symmorphosis concept in sedentary subjects. PMID- 18417646 TI - Loss of prokineticin receptor 2 signaling predisposes mice to torpor. AB - The genes encoding prokineticin 2 polypeptide (Prok2) and its cognate receptor (Prokr2/Gpcr73l1) are widely expressed in both the suprachiasmatic nucleus and its hypothalamic targets, and this signaling pathway has been implicated in the circadian regulation of behavior and physiology. We have previously observed that the targeted null mutation of Prokr2 disrupts circadian coordination of cycles of locomotor activity and thermoregulation. We have now observed spontaneous but sporadic bouts of torpor in the majority of these transgenic mice lacking Prokr2 signaling. During these torpor bouts, which lasted for up to 8 h, body temperature and locomotor activity decreased markedly. Oxygen consumption and carbon dioxide production also decreased, and there was a decrease in respiratory quotient. These spontaneous torpor bouts generally began toward the end of the dark phase or in the early light phase when the mice were maintained on a 12:12-h light-dark cycle and persisted when mice were exposed to continuous darkness. Periods of food deprivation (16-24 h) induced a substantial decrease in body temperature in all mice, but the duration and depth of hypothermia was significantly greater in mice lacking Prokr2 signaling compared with heterozygous and wild-type littermates. Likewise, when tested in metabolic cages, food deprivation produced greater decreases in oxygen consumption and carbon dioxide production in the transgenic mice than controls. We conclude that Prokr2 signaling plays a role in hypothalamic regulation of energy balance, and loss of this pathway results in physiological and behavioral responses normally only detected when mice are in negative energy balance. PMID- 18417647 TI - Rapid cold-hardening in larvae of the Antarctic midge Belgica antarctica: cellular cold-sensing and a role for calcium. AB - In many insects, the rapid cold-hardening (RCH) response significantly enhances cold tolerance in minutes to hours. Larvae of the Antarctic midge, Belgica antarctica, exhibit a novel form of RCH, by which they increase their freezing tolerance. In this study, we examined whether cold-sensing and RCH in B. antarctica occur in vitro and whether calcium is required to generate RCH. As demonstrated previously, 1 h at -5 degrees C significantly increased organismal freezing tolerance at both -15 degrees C and -20 degrees C. Likewise, RCH enhanced cell survival of fat body, Malpighian tubules, and midgut tissue of larvae frozen at -20 degrees C. Furthermore, isolated tissues retained the capacity for RCH in vitro, as demonstrated with both a dye exclusion assay and a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT)-based viability assay, thus indicating that cold-sensing and RCH in B. antarctica occur at the cellular level. Interestingly, there was no difference in survival between tissues that were supercooled at -5 degrees C and those frozen at -5 degrees C, suggesting that temperature mediates the RCH response independent of the freezing of body fluids. Finally, we demonstrated that calcium is required for RCH to occur. Removing calcium from the incubating solution slightly decreased cell survival after RCH treatments, while blocking calcium with the intracellular chelator BAPTA-AM significantly reduced survival in the RCH treatments. The calmodulin inhibitor N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide hydrochloride (W-7) also significantly reduced cell survival in the RCH treatments, thus supporting a role for calcium in RCH. This is the first report implicating calcium as an important second messenger in the RCH response. PMID- 18417648 TI - Modulation of single-nephron GFR in the db/db mouse model of type 2 diabetes mellitus. II. Effects of renal mass reduction. AB - This study examines for the first time the effects of uninephrectomy (Nx) on modulation of whole kidney glomerular filtration rate (GFR), single-nephron GFR (SNGFR), and progression of diabetic nephropathy in the db/db mouse model of type 2 diabetes mellitus. To characterize SNGFR and tubuloglomerular feedback (TGF) responses to Nx and chronic neuronal nitric oxide synthase inhibition in the db/db mouse, we studied the effects of Nx on whole kidney GFR, SNGFR, and TGF characteristics in db/db and wild-type (WT) mice after Nx or sham Nx. We also documented progression of glomerular changes over a 6-mo period. Whole kidney GFR and SNGFR were significantly higher in db/db Nx than db/db sham mice, without change in proximal tubule reabsorptive rates. The TGF responses, determined as proximal-distal SNGFR differences, were brisk: 12.1 +/- 1.0 vs. 8.4 +/- 0.6 nl/min in WT sham (P < 0.05), 15.7 +/- 1.0 vs. 12.0 +/- 1.0 nl/min in WT Nx (P < 0.05), and 17.8 +/- 1.3 vs. 14.3 +/- 1.0 nl/min in db/db Nx (P < 0.05) mice. Chronic ingestion of the neuronal nitric oxide synthase inhibitor S methylthiocitrulline for 2-3 wk after Nx had no effect on SNGFR or the TGF response. These studies show further elevations in whole kidney GFR and SNGFR in these hyperglycemic morbidly obese db/db mice, with an intact TGF system after Nx. In addition, in the db/db Nx mice, 4-6 mo after Nx, there was an exacerbation of the lesions of diabetic nephropathy, as quantified by a significant increase in the ratio of mesangial surface area to total glomerular surface area. PMID- 18417649 TI - Leptin extends the anorectic effects of chronic PYY(3-36) administration in ad libitum-fed rats. AB - Acute administration of peptide YY(3-36) [PYY(3-36)] results in a reduction in food intake in several different vertebrates. However, long-term continuous administration of PYY(3-36) causes only a transient reduction in food intake, thus potentially limiting its therapeutic efficacy. We hypothesized that a fall in leptin levels associated with reduced food intake could contribute to the transient anorectic effects of continuous PYY(3-36) infusion and thus that leptin replacement might prolong the anorectic effects of PYY(3-36). Seven-day administration of 100 microg x kg body wt(-1) x day(-1) PYY(3-36) using osmotic minipumps caused a significant reduction in food intake of ad libitum-fed rats, but only for the first 2 days postimplantation. Circulating levels of leptin were reduced 1 day following continuous infusion of PYY(3-36), and combined leptin infusion at a dose of leptin that had no anorectic effects on its own (100 microg x kg body wt(-1) x day(-1)) prolonged the anorectic actions of PYY(3-36) in ad libitum-fed rats for up to 6 days postimplantation and yielded reduced weight gain compared with either peptide alone. The inhibitory effects of 100 microg x kg body wt(-1) x day(-1) PYY(3-36) on food intake were absent in rats refed after a 24-h fast and substantially reduced at a dose of 1,000 microg x kg body wt(-1) x day(-1) PYY(3-36). Leptin replacement was unable to recover the anorectic effects of PYY(3-36) in fasted rats. Our results suggest that an acute fall in leptin levels is not solely responsible for limiting duration of action of chronic PYY(3-36) infusion, yet chronic coadministration of a subanorectic dose of leptin can extend the anorectic effects of PYY(3-36). PMID- 18417650 TI - Paradoxical effects of endurance training and chronic hypoxia on myofibrillar ATPase activity. AB - This study aimed to determine the changes in soleus myofibrillar ATPase (m ATPase) activity and myosin heavy chain (MHC) isoform expression after endurance training and/or chronic hypoxic exposure. Dark Agouti rats were randomly divided into four groups: control, normoxic sedentary (N; n = 14), normoxic endurance trained (NT; n = 14), hypoxic sedentary (H; n = 10), and hypoxic endurance trained (HT; n = 14). Rats lived and trained in normoxia at 760 mmHg (N and NT) or hypobaric hypoxia at 550 mmHg (approximately 2,800 m) (H and HT). m-ATPase activity was measured by rapid flow quench technique; myosin subunits were analyzed with mono- and two-dimensional gel electrophoresis. Endurance training significantly increased m-ATPase (P < 0.01), although an increase in MHC-I content occurred (P < 0.01). In spite of slow-to-fast transitions in MHC isoform distribution in chronic hypoxia (P < 0.05) no increase in m-ATPase was observed. The rate constants of m-ATPase were 0.0350 +/- 0.0023 s(-1) and 0.047 +/- 0.0050 s(-1) for N and NT and 0.033 +/- 0.0021 s(-1) and 0.038 +/- 0.0032 s(-1) for H and HT. Thus, dissociation between variations in m-ATPase and changes in MHC isoform expression was observed. Changes in fraction of active myosin heads, in myosin light chain isoform (MLC) distribution or in MLC phosphorylation, could not explain the variations in m-ATPase. Myosin posttranslational modifications or changes in other myofibrillar proteins may therefore be responsible for the observed variations in m-ATPase activity. PMID- 18417651 TI - Expression of energy balance regulatory genes in the developing ovine fetal hypothalamus at midgestation and the influence of hyperglycemia. AB - Evidence suggests that the prenatal nutritional environment influences the risk of developing obesity, a major health problem worldwide. It is hypothesized that fetal nutrition influences the developing neuroendocrine hypothalamus, the integrative control center for postnatal energy balance regulation. The present aim was to determine whether relevant hypothalamic genes are expressed in midgestation and whether they are nutritionally (glucose) sensitive at this time. Hypothalami from a cohort of 81-day singleton sheep fetuses, with varying glycemia by virtue of maternal dietary and/or growth hormone treatment, were subject to in situ hybridization analysis for primary orexigenic, anorexigenic, and related receptor genes (term = 147 days, n = 24). Neuropeptide Y, agouti related peptide, proopiomelanocortin (POMC), cocaine- and amphetamine-regulated transcript (CART), and insulin receptor mRNAs were all localized in the hypothalamic arcuate nucleus (ARC) of all fetuses, whereas leptin receptor mRNA was expressed more abundantly in the ventromedial hypothalamic nucleus. ARC expression levels of POMC and CART genes, but none of the other genes, were positively correlated with fetal plasma glucose concentrations. Therefore, key central components of adult energy balance regulation were already present as early as midgestation (equivalent to 22 wk in humans), and two anorexigenic components were upregulated by elevated glycemia. Such changes provide a potential mechanism for the prenatal origins of postnatal energy balance dysregulation and obesity. PMID- 18417652 TI - A perinatal nitric oxide donor increases renal vascular resistance and ameliorates hypertension and glomerular injury in adult fawn-hooded hypertensive rats. AB - Enhancing perinatal nitric oxide (NO) availability persistently reduces blood pressure in spontaneously hypertensive rats. We hypothesize that this approach can be generalized to other models of genetic hypertension, for instance those associated with renal injury. Perinatal exposure to the NO donor molsidomine was studied in fawn-hooded hypertensive (FHH) rats, a model of mild hypertension, impaired preglomerular resistance, and progressive renal injury. Perinatal molsidomine increased urinary NO metabolite excretion at 8 wk of age, i.e., 4 wk after treatment was stopped (P < 0.05). Systolic blood pressure was persistently reduced after molsidomine (42-wk females: 118 +/- 3 vs. 141 +/- 5 and 36-wk males: 139 +/- 4 vs. 158 +/- 4 mmHg; both P < 0.001). Perinatal treatment decreased glomerular filtration rate (P < 0.05) and renal blood flow (P < 0.01) and increased renal vascular resistance (P < 0.05), without affecting filtration fraction, suggesting persistently increased preglomerular resistance. At 4 wk of age natriuresis was transiently increased by molsidomine (P < 0.05). Molsidomine decreased glomerulosclerosis (P < 0.05). Renal blood flow correlated positively with glomerulosclerosis in control (P < 0.001) but not in perinatally treated FHH rats. NO dependency of renal vascular resistance was increased by perinatal molsidomine. Perinatal enhancement of NO availability can ameliorate development of hypertension and renal injury in FHH rats. Paradoxically, glomerular protection by perinatal exposure to the NO donor molsidomine may be due to persistently increased preglomerular resistance. The mechanisms by which increased perinatal NO availability can persistently reprogram kidney function and ameliorate hypertension deserve further study. PMID- 18417653 TI - A purine-selective nucleobase/nucleoside transporter in PK15NTD cells. AB - Nucleoside and nucleobase transporters are important for salvage of purines and pyrimidines and for transport of their analog drugs into cells. However, the pathways for nucleobase translocation in mammalian cells are not well characterized. We identified an Na-independent purine-selective nucleobase/nucleoside transport system in the nucleoside transporter-deficient PK15NTD cells. This transport system has 1,000-fold higher affinity for nucleobases than nucleosides with K(m) values of 2.5 +/- 0.7 microM for [(3)H]adenine, 6.4 +/- 0.5 microM for [(3)H]guanine, 1.1 +/- 0.1 mM for [(3)H]guanosine, and 4.2 +/- 0.5 mM [(3)H]adenosine. The uptake of [(3)H]guanine (0.05 microM) was inhibited by other nucleobases and nucleobase analog drugs (at 0.5-1 mM in the order of potency): 6-mercaptopurine = thioguanine = guanine > adenine >>> thymine = fluorouracil = uracil. Cytosine and methylcytosine had no effect. Nucleoside analog drugs with modification at 2' and/or 5 positions (all at 1 mM) were more potent than adenosine in competing the uptake of [(3)H]guanine: 2-chloro-2'-deoxyadenosine > 2-chloroadenosine > 2'3' dideoxyadenosine = 2'-deoxyadenosine > 5-deoxyadenosine > adenosine. 2-Chloro-2' deoxyadenosine and 2-chloroadenosine inhibited [(3)H]guanine uptake with IC(50) values of 68 +/- 5 and 99 +/- 10 microM, respectively. The nucleobase/nucleoside transporter was resistant to nitrobenzylthioinosine {6-[(4-nitrobenzyl) thiol]-9 beta-D-ribofuranosylpurine}, dipyridamole, and dilazep, but was inhibited by papaverine, the organic cation transporter inhibitor decynium-22 (IC(50) of approximately 1 microM), and by acidic pH (pH = 5.5). In conclusion, we have identified a mammalian purine-selective nucleobase/nucleoside transporter with high affinity for purine nucleobases. This transporter is potentially important for transporting naturally occurring purines and purine analog drugs into cells. PMID- 18417654 TI - Evaluation of new commercial real-time PCR quantification assay for prenatal diagnosis of cytomegalovirus congenital infection. AB - A new commercial real-time human cytomegalovirus (HCMV) PCR kit was evaluated after automated DNA extraction of 153 amniotic fluids in parallel with an in house real-time PCR assay. The commercial kit displayed 100% sensitivity/specificity compared to the "in-house" assay and was suitable for prenatal diagnosis of HCMV congenital infection. PMID- 18417655 TI - Norovirus excretion in an aged-care setting. AB - Norovirus genogroup II excretion during an outbreak of gastroenteritis was investigated in an aged-care facility. Viral shedding peaked in the acute stage of illness and continued for an average of 28.7 days. The viral decay rate was 0.76 per day, which corresponds to a viral half-life of 2.5 days. PMID- 18417656 TI - Identification of a porcine calicivirus related genetically to human sapoviruses. AB - Whether animals may act as reservoirs for human caliciviruses is unclear. By sequence analysis of a short fragment of the RNA-dependent RNA polymerase (RdRp) region, porcine sapovirus (SaV) strains that genetically resemble human SaVs have been detected in piglets, but more-informative sequences (capsid gene) were not available for a precise characterization. In this study, the 3' terminus (the 3' end of open reading frame 1 [ORF1], including the polymerase complex and the complete capsid; ORF2; and the 3' untranslated region) of one such human SaV-like strain, 43/06-18p3/2006/It, was determined, revealing that these viruses are more related genetically to human (47.4 to 54.9% amino acid identity) than to animal (35.2 to 44.7% amino acid identity) SaVs in the capsid gene. In addition, the recombination-prone RdRp-capsid junction region was highly conserved with those of human SaVs of genogroup GI. The presence of porcine viruses similar to human SaVs is a significant finding because of the potential for zoonotic infections or generation of porcine/human recombinants. PMID- 18417657 TI - Rapid differentiation of methicillin-susceptible Staphylococcus aureus from methicillin-resistant S. aureus and MIC determinations by isothermal microcalorimetry. AB - In this study, the use of isothermal microcalorimetry (IMC) for differentiation between methicillin-resistant Staphylococcus aureus (MRSA) and methicillin susceptible S. aureus (MSSA) and MIC determination was evaluated. It was possible to differentiate between MRSA and MSSA within 4 h, whereas the standard method required 24 h. The MICs of cefoxitin were successfully determined for MRSA and MSSA by using IMC. PMID- 18417658 TI - Identification of factors contributing to T-cell toxicity of Staphylococcus aureus clinical isolates. AB - We examined the ability of 206 clinical isolates of Staphylococcus aureus to lyse T cells and found differences between Agr groups. We found that the beta and delta hemolysins are involved and that methicillin-resistant S. aureus strains are less toxic than methicillin-susceptible S. aureus strains. PMID- 18417659 TI - Comparison of DNA pyrosequencing with alternative methods for identification of mycobacteria. AB - Identification of mycobacterial clinical isolates by pyrosequencing within the hypervariable A region of the 16S rRNA gene was compared to other identification methods. For >90% of isolates, these identifications correlated to the level of complex or species. For identification of many mycobacteria, pyrosequencing offers an inexpensive alternative to traditional sequencing. PMID- 18417660 TI - Outbreak of OXY-2-Producing Klebsiella oxytoca in a renal transplant unit. AB - We describe a Klebsiella oxytoca infection outbreak in a renal transplant unit that involved seven patients. All strains belonged to a single pulsed-field gel electrophoresis pattern and were resistant to amoxicillin-clavulanate, cefuroxime, piperacillin-tazobactam, and aztreonam but susceptible to ceftriaxone, ceftazidime, cefepime, and imipenem. Chromosomal beta-lactamase hyperproduction was caused by a point mutation in the bla(OXY-2) gene promoter region. PMID- 18417661 TI - Sequence type and emm type diversity in Streptococcus pyogenes isolates causing invasive disease in Norway between 1988 and 2003. AB - The incidence of invasive group A streptococcal disease has increased in Norway since the 1980s. Analysis of 100 isolates recovered from 1988 to 2003 showed an increased genotypic diversity over time, while the prevalence of the strain that dominated in 1988, sequence type (ST)-28/emm-1, decreased. Necrotizing fasciitis was often associated with ST-15/emm-3. PMID- 18417662 TI - Clostridium difficile toxinotype V, ribotype 078, in animals and humans. PMID- 18417663 TI - Accuracy and potential usefulness of triplex real-time PCR for improving antibiotic treatment of patients with blood cultures showing clustered gram positive cocci on direct smears. AB - Bacterial identification and antibiotic susceptibility testing currently require 48 h when a first blood culture (BC) is positive for clustered gram-positive cocci on direct smear examination (DSE). Meanwhile, antibiotic treatment is often inadequate, reducing the chances of effective treatment or creating unnecessary selective pressure. A new real-time PCR (RT-PCR) technique that differentiates Staphylococcus aureus from coagulase-negative staphylococci (CoNS) and detects methicillin resistance in 90 min in BC bottles could help solve these problems. BC bottles from 410 patients with gram-positive cocci on DSE were processed by current methods, and patients' treatments were prospectively recorded. The RT-PCR assay was performed on aliquots of these BCs, which had been kept frozen. For the 121 patients who had true bacteremia, we established whether the faster availability of RT-PCR results could have led to the initiation of treatments different from those actually given. RT-PCR sensitivity and specificity were 100% for differentiating between S. aureus and CoNS and detecting methicillin resistance with two manufacturers' BC bottles. For 31/86 (36%) of the S. aureus infected patients and for 8/35 (23%) of the CoNS-infected patients who either had suboptimal or nonoptimal treatment or were untreated 48 h after positivity was detected, the early availability of RT-PCR results could have allowed more effective treatment. Unnecessary glycopeptide treatments could have been avoided for 28 additional patients. The use of RT-PCR would increase treatment effectiveness in patients with staphylococcal bacteremia and reduce the selective pressure created by glycopeptides. PMID- 18417664 TI - Changes in tetracycline susceptibility of enteric bacteria following switching to nonmedicated milk replacer for dairy calves. AB - A randomized intervention study was conducted to determine if discontinuing use of calf milk replacer medicated with oxytetracycline results in increased tetracycline susceptibility in Salmonella and Campylobacter spp. and Escherichia coli in dairy calves over a 12-month period. Dairy herds with enteric bacteria with known low tetracycline susceptibility were enrolled for the study. Fecal samples from preweaned calves and environmental samples were collected from eight dairy herds in Michigan and New York State. Samples were collected monthly for 3 months prior to and 12 months after four of the eight herds discontinued medicated milk replacer feeding. Salmonella and Campylobacter spp. and E. coli were isolated, and antimicrobial susceptibility testing was conducted using automated broth microdilution. A total of 804 intervention and 1,026 control calf fecal samples and 122 intervention and 136 control environmental samples were collected for testing. No differences in owner-reported morbidity and mortality between treatment groups were seen. The intervention was significantly associated with increasing tetracycline susceptibility in E. coli and Salmonella. Tetracycline susceptibility increased in intervention herds for the first 3 months after switching to nonmedicated milk replacer but declined in subsequent months. Discontinuing the practice of feeding medicated milk replacers to calves increased tetracycline susceptibility in E. coli and Salmonella on dairy farms, without increasing cattle disease, but declines in effectiveness after 3 months suggest that other factors contribute to decreasing susceptibility on the farm. PMID- 18417665 TI - Characterization of in vitro-generated and clinical optochin-resistant strains of Streptococcus pneumoniae isolated from Argentina. AB - Optochin susceptibility is a key test used for pneumococcal diagnosis, but optochin-resistant (Opt(r)) pneumococci have been reported in the last 2 decades. In this work, we characterized eight Opt(r) clinical strains which presented a new mutation, G47V, a predominant A49S mutation (recently reported in Brazil) and A49T. These mutations were found in the c subunit of the F(0)F(1) ATPase encoded by the atpC gene, and W206C was found in the a subunit encoded by the atpA gene. The Opt(r) clinical isolates were analyzed by BOX PCR, multilocus sequence typing, and serotype and antimicrobial resistance profiles, and they showed no epidemiological relationship. To characterize the Opt(r) mutations that could emerge among clinical strains, we studied a pool of spontaneous Opt(r) colonies obtained in vitro from the virulent D39 strain. We compared the atpAC mutations of these Opt(r) pneumococci (with or without passage through C57BL/6 mice) with those described in the clinical isolates. This analysis revealed three new mutations, G47V and L26M in the c subunit and L184S in the a subunit. Most of the mutations identified in the laboratory-generated Opt(r) strains were also found in clinical strains, with the exception of the L26M and L184S mutations, and we suppose that both mutations could emerge among invasive strains in the future. Considering that atpAC are essential genes, we propose that all spontaneous mutations that confer in vitro optochin resistance would not present severe physiological alterations in S. pneumoniae and may be carried by circulating pneumococcal strains. PMID- 18417666 TI - Significantly improved accuracy of diagnosis of early Lyme disease by peptide enzyme-linked immunosorbent assay based on the borreliacidal antibody epitope of Borrelia burgdorferi OspC. AB - Highly specific borreliacidal antibodies are induced by infection with Borrelia burgdorferi, and the immunodominant response during early Lyme disease is specific for an epitope within the 7 amino acids nearest the C terminus of OspC. We evaluated the ability of an enzyme-linked immunosorbent assay (ELISA) based on a synthetic peptide (OspC7) that matched the region to detect the response and compared the sensitivity during early Lyme disease to that for an FDA-approved Western blot. When the optical density value was adjusted to 98% specificity based on the results from testing normal or uncharacterized sera (n = 236) or sera from patients with blood factors or illnesses that commonly produce antibodies that cross-react with B. burgdorferi antigens (n = 77), 115 (73%) of 157 sera from patients likely to have early Lyme disease were positive for immunoglobulin M (IgM) antibodies and 17 (11%) also had IgG antibodies. In addition, the IgM ELISA reactivities and the titers of antibodies detected by a flow cytometric borreliacidal antibody test correlated closely (r = 0.646). Moreover, the IgM ELISA was significantly more sensitive (P < 0.001) than the Western blot procedure. The findings therefore confirmed that the peptide IgM ELISA detected OspC borreliacidal antibodies and provided strong evidence that the test can eliminate the necessity for confirming early Lyme disease by a supplementary test such as Western blotting. PMID- 18417667 TI - Immunoglobulin G avidities in infants in Mexico after primary immunization with three doses of polyribosylribitol phosphate-tetanus toxoid Haemophilus influenzae type b vaccine. AB - Serum immunoglobulin G concentrations and avidities specific to Haemophilus influenzae type b (Hib) were measured in 208 children living in Guadalajara and Mexico City. Protective concentrations were found in 98.9% and 100.0% of participants, respectively. Geometric mean concentrations differed between both populations and/or among age groups. Mean avidities differed only among the 7- to 12-month-old children. Diphtheria-tetanus-whole-cell pertussis-hepatitis B-Hib primary vaccination seems to induce protection in Mexican children. PMID- 18417668 TI - Interlaboratory comparison of results of an anthrax lethal toxin neutralization assay for assessment of functional antibodies in multiple species. AB - The anthrax lethal toxin neutralization assay (TNA) will likely be used to correlate the protection offered by new anthrax vaccines in animal models to the immunogenicity that will be provided in humans. TNA data are being generated in several different laboratories to measure the immune responses in rabbits, nonhuman primates, and humans. In order to compare data among species and laboratories, a collaborative study was conducted in which 108 samples from the three species were analyzed in seven independent laboratories. Six of the seven laboratories had participated in an interlaboratory technology transfer of the TNA. Analysis of the titration curves generated by samples from each species indicated that the behaviors of the samples from all species were similar; the upper and lower asymptotes and the slopes of the curves were less than 30% divergent from those for human reference material. Dilutional linearity was consistent among samples from each species, with spike to effective dilution at 50% inhibition (ED(50)) slopes of less than 1.2 for all species. Agreement among the laboratories with consensus values was within 10% of the ED(50)s for all samples and within 7.5% of the quotients of the test sample ED(50) and the reference standard ED(50) (NF(50)s) for all samples. The relative standard deviations obtained when data from all laboratories and for all species were combined were 45% for the ED(50)s and 35% for the NF(50)s. These precision data suggest that the NF(50) readout may normalize the values generated by different laboratories. This study demonstrates that the TNA is a panspecies assay that can be performed in several different laboratories with a high degree of quantitative agreement and precision. PMID- 18417670 TI - "Missed" diagnoses of phyllodes tumor on breast biopsy: pathologic clues to its recognition. AB - Fibroadenoma and phyllodes tumors of the breast exhibit a continuum of pathologic features. We examined phyllodes tumors initially called fibroadenoma for features that may accurately classify the tumor as phyllodes tumor on the first biopsy specimen. The phyllodes tumors initially called fibroadenoma for features that may accurately classify the tumor as phyllodes tumors on the first biopsy specimen are examined. Fifteen patients with phyllodes tumors were studied, initially called FA or another term short of PT. These tumors were compared with 16 true fibroadenomas, all with needle-core biopsy followed by excision. Resected phyllodes tumors were larger on average than fibroadenoma, 6.8 cm (range = 1.7 16.2 cm) versus 2.6 cm (range = 1.0-4.8 cm). In needle-core biopsy cases, sampling was limited, even in large breast masses. p53 and cleaved caspase-3 were noncontributory. Ki-67 showed higher proliferation indices in phyllodes tumors versus fibroadenoma (4.8% vs 0.6%). Features suggesting phyllodes tumors include tissue fragmentation, increased stromal cellularity especially around glands, stromal overgrowth, and increased mitoses. Increased sampling of a large tumor will likely yield more correct diagnoses. PMID- 18417671 TI - OLGA gastritis staging in young adults and country-specific gastric cancer risk. AB - Geographical differences have been shown in the clinical outcomes of Helicobacter pylori-associated gastritis phenotypes and in gastric cancer risk. This study tested whether the Operative Link on Gastritis Assessment (OLGA) staging correlated with gastric cancer risk in populations from 3 continents. Mapped gastric biopsies were obtained from 316 dyspeptic adults aged less than 41 years from 8 geographic areas that differed in gastric cancer risk. Gastric atrophy was assessed according to internationally validated criteria. Gastritis stage was established according to the OLGA staging system. The most prevalent gastritis stages were 0 to II, which included all subjects entered from Chile, Germany, India, Italy, and Thailand. Gastritis Stages III and IV were limited to the Chinese and Korean populations. Indians had a high prevalence of H pylori infection without high-stage gastritis. In populations at different cancer risk, the gastritis OLGA stage mirrored the gastric cancer incidence. Gastritis staging identifies a subgroup of higher-risk patients. PMID- 18417672 TI - Breast carcinoma with micropapillary features: clinicopathologic study and long term follow-up of 100 cases. AB - To study the clinicopathologic characteristics and prognosis of invasive micropapillary carcinoma of breast (IMPC), 100 cases of invasive breast carcinoma with an IMPC component were reviewed. Compared with invasive ductal carcinoma, not otherwise specified, with similar histologic grades, carcinomas with IMPC were larger sized, had a higher lymph node metastasis rate with more nodes involved per case, and exhibited increased lymphovascular invasion. The presence of IMPC strongly correlated with the more aggressive behavior. No significant association was established between the proportion of the IMPC component and overall tumor size, histologic grade, lymph node metastasis rate, and distant metastasis, but a trend was noted. Long-term follow-up demonstrated a poorer 5 year and 10-year survival rate for patients with breast carcinoma containing an IMPC component. Breast carcinomas with micropapillary features are more aggressive tumors with a poorer prognosis. This specific structure should be carefully evaluated in the surgical pathology examination of breast carcinoma specimens. PMID- 18417673 TI - Marginal zone B-Cell lymphoma among primary B-Cell lymphoma of Waldeyer's ring: histopathologic and immunohistochemical study of 16 tonsillectomy specimens. AB - Two subtypes of marginal zone B-cell lymphoma (eg, mucosa-associated lymphoid tissue [MALT] type and splenic type) have been reported in the lymph node. To determine the presence or absence of marginal zone B-cell lymphoma of MALT type and the splenic type among Waldeyer's ring (WR) lymphomas, 16 tonsillectomy specimens were studied. Ten cases (63%) were marginal zone B-cell lymphoma. Among marginal zone B-cell lymphoma, 7 were the MALT type and the remaining 3 cases of marginal zone B-cell lymphoma were the splenic type. Moreover, 4 cases of 7 MALT type lymphomas contained numerous large cells (diffuse large B-cell lymphoma arising from a low-grade marginal zone B-cell lymphoma of MALT type). The low incidence of primary mucosa-associated lymphoid tissue type lymphoma of WR in previous reports may be because it is difficult to correctly identify the characteristic histologic findings of MALT-type lymphoma because of the small biopsy size. PMID- 18417674 TI - Bilateral lipomastia in men: a side effect of highly active antiretroviral therapy. AB - Breast enlargement in men and women is a documented feature of the highly active antiretroviral therapy- associated human immunodeficiency virus lipodystrophy syndrome. The exact underlying histomorphological features of this condition are speculative because most cases are diagnosed on clinical grounds with or without radiographic confirmation. The main documented causes of breast enlargement in men on highly active antiretroviral therapy include gynecomastia and lipomastia; however, biopsy-confirmed lipomastia is a rarely described phenomenon, with only 1 such case being described to date. In documenting 2 patients who underwent bilateral mastectomy for clinical gynecomastia of unknown cause, we emphasize the need for a greater degree of clinicopathological awareness of highly active antiretroviral therapy-associated lipomastia and the role of the histopathologist in the informed management of patients afflicted with human immunodeficiency virus infection on highly active antiretroviral therapy. PMID- 18417675 TI - Pseudo-Gaucher cells in multiple myeloma. AB - A case of multiple myeloma is reported in which the bone marrow contained sheets of histiocytes with light microscopic features mimicking Gaucher cells. The patient had no clinical evidence of inherited Gaucher's disease. These pseudo Gaucher cells obscured neoplastic plasma cells causing diagnostic difficulty. PMID- 18417676 TI - Ovarian signet-ring stromal tumor: a potential diagnostic pitfall. AB - Signet-ring stromal tumor is a rare ovarian neoplasm with only 10 reported cases in the literature. We report an unusual case of ovarian signet-ring stromal tumor in a 69-year-old woman who presented with right adnexal mass and underwent total abdominal hysterectomy and bilateral salpingo-oophorectomy. The diagnosis was based on histological, histochemical, immunohistochemical, and electron microscopy characteristics. The main significance is to differentiate this benign tumor from the highly malignant Krukenberg tumor, and this differential diagnosis is discussed. PMID- 18417678 TI - Docetaxel-induced mitotic arrest in epithelium of gallbladder: a hitherto unreported occurrence. AB - Chemotherapeutic agents that bind tubulin cause mitotic arrest, which may be seen histologically. Such mitotic arrest has been reported to occur in the skin, alimentary canal, lungs, liver, bone marrow, endometrium, breasts, or in ascites following treatment with paclitaxel, vincristine, colchicine, podophyllotoxin, or maytansine. Mitotic arrest as a result of docetaxel, a taxane that binds tubulin, has yet to be reported. Mitotic arrest in the gallbladder has also yet to be reported. We recently encountered a case of dramatic mitotic arrest as a result of docetaxel, involving the gallbladder of a 66-year-old man with metastatic bronchogenic carcinoma. Strikingly abundant bizarre mitoses initially prompted a diagnosis of primary carcinoma. Carcinoma was eventually excluded based on the absence of dysplasia in all cells at interphase and the history of recent administration of docetaxel. This is the first case of mitotic arrest involving docetaxel or the gallbladder. Awareness of this phenomenon is necessary to avoid misdiagnosing carcinoma. PMID- 18417679 TI - Intestinal clear cell sarcoma with melanocytic differentiation and EWS [corrected] rearrangement: report of a case. AB - A clear cell sarcoma-like tumor with osteoclast-like giant cells of the gastrointestinal tract without immunoexpression of CD117 was recently proposed as a new tumor entity. In this article, a case of a 37-year-old man with a neoplasm of the jejunum composed of polygonal cells with clear to eosinophilic cytoplasm forming nests and fascicles is reported. Giant cells were not identified. Immunohistochemically, the tumor cells expressed strongly S100 protein, human melanoma black 45, platelet-derived growth factor receptor alpha, B-cell lymphoma 2, p53, and to a lesser extent vimentin, neuron-specific enolase, and epithelial membrane antigen. Mindbomb homolog-1 index was 35%. Immunoreactivity for CD34 and CD117 was negative. The fluorescence in situ hybridization analysis showed a translocation involving chromosome 22q12, the diagnostic hallmark of clear cell sarcoma of soft tissues. This case indicates a close histogenetic relationship with the recently reported clear cell sarcoma with osteoclast-like giant cells of the gastrointestinal tract, as well as with the clear cell sarcoma of soft tissues. PMID- 18417680 TI - Aberrant expression of alpha-fetoprotein in intrahepatic cholangiocarcinoma: an exceptional occurrence. AB - A high level of serum alpha-fetoprotein (AFP) is typically indicative of hepatocellular carcinoma in patients with liver lesions. In this article, we describe an exceptional case of intrahepatic cholangiocarcinoma that occurred in a 36-year-old man with a markedly elevated serum AFP level (12310.7 ng/mL). Histopathologic examination of surgically resected liver mass showed classic morphologic features of cholangiocarcinoma, with no hepatocellular carcinoma component identified. Immunohistochemically, the tumor cells were strongly and diffusely positive for AFP, CA19-9, and cytokeratin 19 and were negative for hepatocyte antigen. The patient's serum AFP level declined to 46.2 ng/mL 1 month after surgery. To the best of our knowledge, this is the first documented case of AFP-producing intrahepatic cholangiocarcinoma with immunohistochemical evidence confirming the tumor cells to be the source of high-level AFP production. PMID- 18417681 TI - Basaloid squamous cell carcinoma arising in [corrected] the renal pelvis. AB - Basaloid squamous cell carcinoma is an uncommon and highly aggressive variant that can occur anywhere squamous cancers occur. It is most often seen in the head and neck, the perianal region, and the female genital tract. It is extremely rare in the urinary system. In this article, we report the first known case of basaloid squamous cell carcinoma arising in the renal pelvis. Given the aggressive nature of this neoplasm, it should not be omitted from the differential diagnosis of neoplasms arising from the urothelium. PMID- 18417682 TI - Acinic cell carcinoma of salivary gland with massive deposits of globular amyloid. AB - We present the case of a 23-year-old woman with a parotid gland tumor, the fine needle aspiration biopsy smears of which showed epithelial cells with wide cytoplasm, isolated or arranged in micropapillary groups together with psammoma bodies. The surgical specimen contained a 5-cm tumor with the histologic features of an acinic cell carcinoma (ACC) with papillary areas. Notably, the cells of the tumor seemed to follow a sequence from large cells with rounded nuclei with open chromatin and prominent nucleoli to vacuolated cells with granular material, and finally to cells undergoing apoptosis. This finding was followed by the appearance of concentrically laminated, round to polygonal, Congo red-positive, birefringent bodies that in areas accumulated and formed extensive areas with massive deposits. The picture suggested that those amyloid bodies (psammoma bodies) resulted from the accumulation of residual masses of apoptotic cells. Huge globular amyloid deposits, the suggested name for this material irrespective of the type of amyloid, have not been previously reported in ACC of salivary gland. PMID- 18417683 TI - Cystic hypersecretory carcinoma of the breast with paget disease of the nipple: a diagnostic challenge. AB - A unique case of cystic hypersecretory carcinoma in a 48-year-old woman who presented with a tender, palpable abnormality of the lower outer quadrant of her left breast in 2002 is described. Mammographic and ultrasound examinations showed heterogeneous, dense breast tissue with a focal asymmetric density and an ill defined heterogenous hypoechoic area with anechoic spaces, respectively, that corresponded to the area of palpable abnormality. Although the imaging findings did not change significantly over 4 years, the patient complained of intermittent, spontaneous serous discharge from the left nipple and persistent induration and tenderness in the same area of the left breast. An ultrasound guided core-needle biopsy of the lesion was performed at the last visit to rule out a malignant process. A left simple mastectomy showed a large multicystic lesion occupying most of the lower quadrants by a cystic hypersecretory carcinoma and Paget disease of the nipple. PMID- 18417684 TI - CD138-positive plasmacytoid urothelial carcinoma of the bladder. PMID- 18417685 TI - Alpha-1-antitrypsin-positive "signet-ring" bile duct adenoma in a patient with M(MALTON) mutation. AB - A 65-year-old man with a colonic adenocarcinoma and a single small liver nodule underwent surgery. The nodule showed the histological features of a bile duct adenoma. Large eosinophilic, periodic acid-Schiff-positive diastase-resistant intracytoplasmic globules conferred a signet-ring appearance to the cells. Similar globules were found in the biliary epithelium in perinodular portal tracts, whereas smaller globules were detected in periportal hepatocytes. All globules showed strong immunoreactivity for alpha-1-antitrypsin, and the sequence of the alpha-1-antitrypsin gene showed a heterozygous M(malton) genotype. The selective recurrence of large alpha-1-antitrypsin globules within the bile ducts may indicate a neoplastic rather than a reactive or hamartomatous nature of the nodule. PMID- 18417686 TI - Wolffian tumor of the ovary with a prominent spindle cell component: report of a case with brief discussion of unusual problems in differential diagnosis, and literature review. AB - An 87-year-old woman underwent total abdominal hysterectomy and bilateral salpingo-oophorectomy for endometrioid adenocarcinoma. At operation, a 4.5-cm partially solid and partially cystic right ovarian mass was identified. Frozen section showed a cytologically bland spindle cell proliferation reminiscent of cellular fibroma. Extensive sampling of the ovarian mass revealed a focus with the classic sieve-like pattern of a Wolffian adnexal tumor that merged with the fibroma-like appearance. The uterus showed well-differentiated, superficially invasive endometrioid adenocarcinoma arising in a background of atypical complex hyperplasia. Given the recent reports implying a therapeutic value for Gleevec (Novartis, Stein, Switzerland) (STI-571) in the treatment of Wolffian tumor of the ovary, paraffin immunostain for CD117 (c-kit) was performed that yielded negative results. Without further therapy, the patient was alive without disease 7 months after surgery. This case demonstrates the heterogeneity of Wolffian tumor of the ovary and shows how crucial sampling is in arriving at the correct diagnosis. PMID- 18417688 TI - The NIH Public Access Policy. PMID- 18417687 TI - Uterine gestational choriocarcinoma developing after a long latent period in a postmenopausal woman: the value of DNA polymorphism studies. AB - This article reports a uterine gestational choriocarcinoma arising in a 57-year old woman with a long latent period of 22 years from the last known pregnancy. Diagnosis was made on an endometrial biopsy specimen, and given the age of the patient, the long latent period, and the limited sample, trophoblastic differentiation within an endometrial carcinoma was considered. The results of DNA polymorphism studies illustrated both paternal and maternal alleles within the tumor in equal amounts, confirming the neoplasm to be gestational in origin and to have originated from a nonmolar gestation. The report discusses the value of DNA polymorphism studies in distinguishing gestational from nongestational choriocarcinoma and from trophoblastic differentiation within a carcinoma. PMID- 18417689 TI - Subjective experience of motion or attentional selection of a categorical decision. PMID- 18417690 TI - Anticipatory brain activity in irritable bowel syndrome. PMID- 18417691 TI - Mitochondrial dysfunction in SOD1G93A-bearing astrocytes promotes motor neuron degeneration: prevention by mitochondrial-targeted antioxidants. AB - Mitochondrial dysfunction and oxidative stress contribute to motor neuron degeneration in amyotrophic lateral sclerosis (ALS). Recent reports indicate that astrocytes expressing the mutations of superoxide dismutase-1 (SOD1) may contribute to motor neuron injury in ALS. Here, we provide evidence that mitochondrial dysfunction in SOD1(G93A) rat astrocytes causes astrocytes to induce apoptosis of motor neurons. Mitochondria from SOD1(G93A) rat astrocytes displayed a defective respiratory function, including decreased oxygen consumption, lack of ADP-dependent respiratory control, and decreased membrane potential. Protein 3-nitrotyrosine was detected immunochemically in mitochondrial proteins from SOD1(G93A) astrocytes, suggesting that mitochondrial defects were associated with nitroxidative damage. Furthermore, superoxide radical formation in mitochondria was increased in SOD1(G93A) astrocytes. Similar defects were found in mitochondria isolated from the spinal cord of SOD1(G93A) rats, and pretreatment of animals with the spin trap 5,5-dimethyl-1-pyrroline N-oxide restored mitochondrial function, forming adducts with mitochondrial proteins in vivo. As shown previously, SOD1(G93A) astrocytes induced death of motor neurons in cocultures, compared with nontransgenic ones. This behavior was recapitulated when nontransgenic astrocytes were treated with mitochondrial inhibitors. Remarkably, motor neuron loss was prevented by preincubation of SOD1(G93A) astrocytes with antioxidants and nitric oxide synthase inhibitors. In particular, low concentrations (approximately 10 nm) of two mitochondrial-targeted antioxidants, ubiquinone and carboxy-proxyl nitroxide, each covalently coupled to a triphenylphosphonium cation (Mito-Q and Mito-CP, respectively), prevented mitochondrial dysfunction, reduced superoxide production in SOD1(G93A) astrocytes, and restored motor neuron survival. Together, our results indicate that mitochondrial dysfunction in astrocytes critically influences motor neuron survival and support the potential pharmacological utility of mitochondrial targeted antioxidants in ALS treatment. PMID- 18417692 TI - Neurturin-mediated ret activation is required for retinal function. AB - The glial cell line-derived neurotrophic factor (GDNF) family ligands (GFLs) [GDNF, NRTN (neurturin), ARTN (artemin), and PSPN (persephin)] interact with GDNF family receptors (GFRalphas) and activate intracellular signaling through the Ret receptor tyrosine kinase. To characterize the role of Ret signaling in retinal activity, we examined Ret hypomorphic and Ret conditional mice using electroretinography. We found that aberrant Ret function resulted in markedly diminished scotopic and photopic responses. Using mice deficient in individual GFLs, we found that only NRTN deficiency led to reduced retinal activity. To determine the potential target cell type for NRTN, we examined the retinal expression of its coreceptors (GFRalpha1 and GFRalpha2) and Ret using mice expressing fluorescence reporter enhanced green fluorescent protein from their respective loci. We found robust GFRalpha1 and Ret expression in horizontal, amacrine, and ganglion cells, whereas GFRalpha2 expression was only detected in a subset of amacrine and ganglion cells. In contrast to previous studies, no expression of GFRalpha1, GFRalpha2, or Ret was detected in photoreceptors or Muller cells, suggesting that these cells are not directly affected by Ret. Finally, detailed morphologic analyses of retinas from NRTN- and Ret-deficient mice demonstrated a reduction in normal horizontal cell dendrites and axons, abnormal extensions of horizontal cell and bipolar cell processes into the outer nuclear layer, and mislocalized synaptic complexes. These anatomic abnormalities indicate a possible basis for the abnormal retinal activity in the Ret and NRTN mutant mice. PMID- 18417693 TI - Disinhibition combines with excitation to extend the operating range of the OFF visual pathway in daylight. AB - Cone signals divide into parallel ON and OFF bipolar cell pathways, which respond to objects brighter or darker than the background and release glutamate onto the corresponding type of ganglion cell. It is assumed that ganglion cell excitatory responses are driven by these bipolar cell synapses. Here, we report an additional mechanism: OFF ganglion cells were driven in part by the removal of synaptic inhibition (disinhibition). The disinhibition played a relatively large role in driving responses at low contrasts. The disinhibition persisted in the presence of CNQX and d-AP-5. Furthermore, the CNQX/d-AP-5-resistant response was blocked by l-AP-4, meclofenamic acid, quinine, or strychnine but not by bicuculline. Thus, the disinhibition circuit was driven by the ON pathway and required gap junctions and glycine receptors but not ionotropic glutamate or GABA(A) receptors. These properties implicate the AII amacrine cell, better known for its role in rod vision, as a critical circuit element through the following pathway: cone --> ON cone bipolar cell --> AII cell --> OFF ganglion cell. Rods could also drive this circuit through their gap junctions with cones. Thus, to light decrement, AII cells, driven by electrical synapses with ON cone bipolar cells, would hyperpolarize and reduce glycine release to excite OFF ganglion cells. To light increment, the AII circuit would directly inhibit OFF ganglion cells. These results show a new role for disinhibition in the retina and suggest a new role for the AII amacrine cell in daylight vision. PMID- 18417694 TI - Conical electron tomography of a chemical synapse: polyhedral cages dock vesicles to the active zone. AB - In this study, we tested the hypothesis that the structure of the active zone of chemical synapses has remained uncertain because of limitations of conventional electron microscopy. To resolve these limitations, we reconstructed chemical synapses of rat neocortex, the archetypical "average" synapse, by conical electron tomography, a method that exhibits an isotropic in plane resolution of approximately 3 nm and eliminates the need to impose symmetry or use averaging methods to increase signal-to-noise ratios. Analysis of 17 reconstructions by semiautomatic density segmentation indicated that the active zone was constructed of a variable number of distinct "synaptic units" comprising a polyhedral cage and a corona of approximately seven vesicles. The polyhedral cages measured approximately 60 nm in diameter, with a density of approximately 44/microm2 and were associated with vesicles at the active zone ("first tier"). Vesicles in this first-tier position represented approximately 7.5% of the total number of vesicles in the terminal and were contiguous, hemifused (approximately 4% of total), or fully fused (approximately 0.5% of total) to the plasma membrane. Our study supports the hypothesis that rat neocortical synapses are constructed of variable numbers of distinct synaptic units that facilitate the docking of vesicles to the active zone and determine the number of vesicles available for immediate release. PMID- 18417695 TI - Silencing the Kir4.1 potassium channel subunit in satellite glial cells of the rat trigeminal ganglion results in pain-like behavior in the absence of nerve injury. AB - Growing evidence suggests that changes in the ion buffering capacity of glial cells can give rise to neuropathic pain. In the CNS, potassium ion (K+) buffering is dependent on the glia-specific inward rectifying K+ channel Kir4.1. We recently reported that the satellite glial cells that surround primary sensory neurons located in sensory ganglia of the peripheral nervous system also express Kir4.1, whereas the neurons do not. In the present study, we show that, in the rat trigeminal ganglion, the location of the primary sensory neurons for face sensation, specific silencing of Kir4.1 using RNA interference leads to spontaneous and evoked facial pain-like behavior in freely moving rats. We also show that Kir4.1 in the trigeminal ganglion is reduced after chronic constriction injury of the infraorbital nerve. These findings suggests that neuropathic pain can result from a change in expression of a single K+ channel in peripheral glial cells, raising the possibility of targeting Kir4.1 to treat pain in general and particularly neuropathic pain that occurs in the absence of nerve injury. PMID- 18417696 TI - Ischemic preconditioning targets the respiration of synaptic mitochondria via protein kinase C epsilon. AB - In the brain, ischemic preconditioning (IPC) diminishes mitochondrial dysfunction after ischemia and confers neuroprotection. Activation of epsilon protein kinase C (epsilonPKC) has been proposed to be a key neuroprotective pathway during IPC. We tested the hypothesis that IPC increases the levels of epsilonPKC in synaptosomes from rat hippocampus, resulting in improved synaptic mitochondrial respiration. Preconditioning significantly increased the level of hippocampal synaptosomal epsilonPKC to 152% of sham-operated animals at 2 d of reperfusion, the time of peak neuroprotection. We tested the effect of epsilonPKC activation on hippocampal synaptic mitochondrial respiration 2 d after preconditioning. Treatment with the specific epsilonPKC activating peptide, tat-psiepsilonRACK (tat-psiepsilon-receptor for activated C kinase), increased the rate of oxygen consumption in the presence of substrates for complexes I, II, and IV to 157, 153, and 131% of control (tat peptide alone). In parallel, we found that epsilonPKC activation in synaptosomes from preconditioned animals resulted in altered levels of phosphorylated mitochondrial respiratory chain proteins: increased serine and tyrosine phosphorylation of 18 kDa subunit of complex I, decreased serine phosphorylation of FeS protein in complex III, increased threonine phosphorylation of COX IV (cytochrome oxidase IV), increased mitochondrial membrane potential, and decreased H2O2 production. In brief, ischemic preconditioning promoted significant increases in the level of synaptosomal epsilonPKC. Activation of epsilonPKC increased synaptosomal mitochondrial respiration and phosphorylation of mitochondrial respiratory chain proteins. We propose that, at 48 h of reperfusion after ischemic preconditioning, epsilonPKC is poised at synaptic mitochondria to respond to ischemia either by direct phosphorylation or activation of the epsilonPKC signaling pathway. PMID- 18417697 TI - Neural measures of individual differences in selecting and tracking multiple moving objects. AB - Attention can be divided so that multiple objects can be tracked simultaneously as they move among distractors. Although attentional tracking is known to be highly limited, such that most individuals can track only approximately four objects simultaneously, the neurophysiological mechanisms that underlie this capacity limitation have not been established. Here, we provide electrophysiological measures in humans of the initial selection and sustained attention processes that facilitate attentional tracking. Each measure was modulated by the number of objects the subject was tracking and was highly sensitive to each individual's specific tracking capacity. Consequently, these measures provide strong neurophysiological predictors of an individual's attentional tracking capacity. Moreover, by manipulating the difficulty of these two phases of the task, we observe that the limiting factor underlying tracking capacity can flexibly shift between these two attentional mechanisms depending on the requirements of the task. PMID- 18417698 TI - Dopamine activates the motor pattern for crawling in the medicinal leech. AB - Locomotion in segmented animals is thought to be based on the coupling of "unit burst generators," but the biological nature of the unit burst generator has been revealed in only a few animal systems. We determined that dopamine (DA), a universal modulator of motor activity, is sufficient to activate fictive crawling in the medicinal leech, and can exert its actions within the smallest division of the animal's CNS, the segmental ganglion. In the entire isolated nerve cord or in the single ganglion, DA induced slow antiphasic bursting (approximately 15 s period) of motoneurons known to participate in the two-step elongation contraction cycle underlying crawling behavior. During each cycle, the dorsal (DE 3) and ventral (VE-4) longitudinal excitor motoneurons fired approximately 180 degrees out of phase from the ventrolateral circular excitor motoneuron (CV), which marks the elongation phase. In many isolated whole nerve cords, DE-3 bursting progressed in an anterior to posterior direction with intersegmental phase delays appropriate for crawling. In the single ganglion, the dorsal (DI-1) and ventral (VI-2) inhibitory longitudinal motoneurons fired out of phase with each DE-3 burst, further confirming that the crawl unit burst generator exists in the single ganglion. All isolated ganglia of the CNS were competent to produce DA induced robust fictive crawling, which typically lasted uninterrupted for 5-15 min. A quantitative analysis indicated that DA-induced crawling was not significantly different from electrically evoked or spontaneous crawling. We conclude that DA is sufficient to activate the full crawl motor program and that the kernel for crawling resides within each segmental ganglion. PMID- 18417699 TI - Dissociation of metabolic and neurovascular responses to levodopa in the treatment of Parkinson's disease. AB - We compared the metabolic and neurovascular effects of levodopa (LD) therapy for Parkinson's disease (PD). Eleven PD patients were scanned with both [15O]-H2O and [18F]-fluorodeoxyglucose positron emission tomography in the unmedicated state and during intravenous LD infusion. Images were used to quantify LD-mediated changes in the expression of motor- and cognition-related PD covariance patterns in scans of cerebral blood flow (CBF) and cerebral metabolic rate for glucose (CMR). These changes in network activity were compared with those occurring during subthalamic nucleus (STN) deep brain stimulation (DBS), and those observed in a test-retest PD control group. Separate voxel-based searches were conducted to identify individual regions with dissociated treatment-mediated changes in local cerebral blood flow and metabolism. We found a significant dissociation between CBF and CMR in the modulation of the PD motor-related network by LD treatment (p < 0.001). This dissociation was characterized by reductions in network activity in the CMR scans (p < 0.003) occurring concurrently with increases in the CBF scans (p < 0.01). Flow-metabolism dissociation was also evident at the regional level, with LD-mediated reductions in CMR and increases in CBF in the putamen/globus pallidus, dorsal midbrain/pons, STN, and ventral thalamus. CBF responses to LD in the putamen and pons were relatively greater in patients exhibiting drug-induced dyskinesia. In contrast, flow-metabolism dissociation was not present in the STN DBS treatment group or in the PD control group. These findings suggest that flow-metabolism dissociation is a distinctive feature of LD treatment. This phenomenon may be especially pronounced in patients with LD-induced dyskinesia. PMID- 18417700 TI - Gray matter increase induced by practice correlates with task-specific activation: a combined functional and morphometric magnetic resonance imaging study. AB - The neurophysiological basis of practice-induced gray matter increase is unclear. To study the relationship of practice-induced gray matter changes and neural activation, we conducted a combined longitudinal functional and morphometric (voxel-based morphometry) magnetic resonance imaging (MRI) study on mirror reading. Compared with normal reading, mirror reading resulted in an activation of the dorsolateral occipital cortex, medial occipital cortex, superior parietal cortex, medial and dorsolateral prefrontal cortex, as well as anterior insula and cerebellum. Daily practice of 15 min for 2 weeks resulted in an increased performance of mirror reading. After correction for pure performance effects, we found a practice-related decrease of activation at the right superior parietal cortex and increase of activation at the right dorsal occipital cortex. The longitudinal voxel-based morphometry analysis yielded an increase of gray matter in the right dorsolateral occipital cortex that corresponded to the peak of mirror-reading-specific activation. This confirms that short-term gray matter signal increase corresponds to task-specific processing. We speculate that practice-related gray matter signal changes in MRI are primarily related to synaptic remodeling within specific processing areas. PMID- 18417701 TI - Acute and chronic dopamine receptor stimulation modulates AMPA receptor trafficking in nucleus accumbens neurons cocultured with prefrontal cortex neurons. AB - Postsynaptic interactions between dopamine (DA) and glutamate receptors in the nucleus accumbens (NAc) are critical for addiction. To determine the effect of acute and repeated DA receptor stimulation on AMPA receptor (AMPAR) synaptic targeting in medium spiny NAc neurons, we developed a model system consisting of rat NAc neurons cocultured with prefrontal cortex neurons from enhanced green fluorescent protein-expressing mice. Cortical neurons restore excitatory input onto NAc neurons but are distinguishable based on fluorescence. First, we showed that brief D1-like agonist exposure increased AMPAR insertion onto extrasynaptic regions of NAc neuronal processes through a mechanism requiring protein kinase A. This facilitated the Ca2+/calmodulin dependent protein kinase II (CaMKII) dependent synaptic incorporation of AMPARs in response to subsequent NMDA receptor (NMDAR) stimulation. Through this mechanism, DA may promote reward- and drug-related plasticity in the NAc. Then, to model effects of repeated in vivo cocaine exposure, we treated cocultures with DA (1 microm, 30 min) on days 7, 9, and 11 in culture. On day 15, NAc neurons exhibited increased synaptic AMPAR levels. This was associated with CaMKII activation and was blocked by the CaMKII inhibitor KN-93 (N-[2-[N-(4-chlorocinnamyl)-N-methylaminomethyl]phenyl]-N-(2 hydroxyethyl)-4-methoxybenzenesulfonamide phosphate salt). Furthermore, D1-like agonist exposure on day 15 no longer increased AMPAR surface expression. This refractoriness was associated with decreased D1 receptor surface expression. NMDAR surface expression was not altered by acute or repeated DA receptor stimulation. These results suggest that (1) after repeated DA treatment, NAc neurons are more responsive to glutamate inputs but D(1)-like receptor regulation of plasticity is impaired, and (2) NAc/prefrontal cortex cocultures are useful for studying dopamine-induced neuroadaptations. PMID- 18417703 TI - Intelligence and variability in a simple timing task share neural substrates in the prefrontal white matter. AB - General intelligence is correlated with the mean and variability of reaction time in elementary cognitive tasks, as well as with performance on temporal judgment and discrimination tasks. This suggests a link between the temporal accuracy of neural activity and intelligence. However, it has remained unclear whether this link reflects top-down mechanisms such as attentional control and cognitive strategies or basic neural properties that influence both abilities. Here, we investigated whether millisecond variability in a simple, automatic timing task, isochronous tapping, correlates with intellectual performance and, using voxel based morphometry, whether these two tasks share neuroanatomical substrates. Stability of tapping and intelligence were correlated and related to regional volume in overlapping right prefrontal white matter regions. These results suggest a bottom-up explanation of the link between temporal stability and intellectual performance, in which more extensive prefrontal connectivity underlies individual differences in both variables. PMID- 18417702 TI - Amyloid beta protein dimer-containing human CSF disrupts synaptic plasticity: prevention by systemic passive immunization. AB - The current development of immunotherapy for Alzheimer's disease is based on the assumption that human-derived amyloid beta protein (Abeta) can be targeted in a similar manner to animal cell-derived or synthetic Abeta. Because the structure of Abeta depends on its source and the presence of cofactors, it is of great interest to determine whether human-derived oligomeric Abeta species impair brain function and, if so, whether or not their disruptive effects can be prevented using antibodies. We report that untreated ex vivo human CSF that contains Abeta dimers rapidly inhibits hippocampal long-term potentiation in vivo and that acute systemic infusion of an anti-Abeta monoclonal antibody can prevent this disruption of synaptic plasticity. Abeta monomer isolated from human CSF did not affect long-term potentiation. These results strongly support a strategy of passive immunization against soluble Abeta oligomers in early Alzheimer's disease. PMID- 18417704 TI - Robos and slits control the pathfinding and targeting of mouse olfactory sensory axons. AB - Odorants are detected by olfactory receptor neurons (ORNs) located in the olfactory epithelium. In mice, ORNs expressing the same odorant receptor (OR) project to a single glomerulus out of 1800 in the olfactory bulb (OB). It has been proposed that OR-derived cAMP signals guide ORN axons to their glomeruli rather than OR themselves. Recently, it has also been shown that the axon guidance molecule Slit1 and its receptor Robo2 control the dorsoventral segregation of ORN axons as they are projecting to the OB. We have analyzed the development of olfactory projections in Slit1/Slit2 and Robo1/Robo2 single and double mutants. We show that in Robo1-/-;Robo2-/- mice, most ORN axons fail to enter the OB and instead project caudally into the diencephalon. Moreover, in these mice, ORN axons expressing the same OR project to several glomeruli at ectopic positions. Thus, Slit1, Slit2, Robo1, and Robo2 cooperate to control the convergence of ORN axons to the OB and the precise targeting of ORN axons to specific glomeruli. PMID- 18417705 TI - Alpha-synuclein alters Notch-1 expression and neurogenesis in mouse embryonic stem cells and in the hippocampus of transgenic mice. AB - Altered expression and mutations in alpha-synuclein (alpha-syn) have been linked to Parkinson's disease (PD) and related disorders. The neurological alterations in PD patients have been associated with degeneration of dopaminergic cells and other neuronal populations. Moreover, recent studies in murine models have shown that alterations in neurogenesis might also contribute to the neurodegenerative phenotype. However, the mechanisms involved and the effects of alpha-syn expression on neurogenesis are not yet clear. To this end, murine embryonic stem (mES) cells were infected with lentiviral (LV) vectors expressing wild-type (WT) and mutant alpha-syn. Compared with mES cells infected with LV-green fluorescent protein (GFP), cells expressing WT and mutant alpha-syn showed reduced proliferation as indicated by lower 5-bromo-2'-deoxyuridine uptake, increased apoptosis, and reduced expression of neuronal markers such as neuron specific enolase and beta-III tubulin. The alterations in neurogenesis in alpha-syn expressing mES cells were accompanied by a reduction in Notch-1 and Hairy and enhancer of split-5 (Hes-5) mRNA and protein levels. Moreover, levels of total Notch-1 and Notch intracellular domain (NICD) were lower in mES cells expressing WT and mutant alpha-syn compared with GFP controls. The reduced survival of alpha syn-expressing mES cells was reverted by overexpressing constitutively active NICD. Similarly, in alpha-syn transgenic mice, the alterations in neurogenesis in the hippocampal subgranular zone were accompanied by decreased Notch-1, NICD, and Hes-5 expression. Together, these results suggest that accumulation of alpha-syn might impair survival of NPCs by interfering with the Notch signaling pathway. Similar mechanisms could be at play in PD and Lewy body disease. PMID- 18417706 TI - Phosphatidylinositol 3-kinase is a key mediator of central sensitization in painful inflammatory conditions. AB - Here, we show that phosphatidylinositol 3-kinase (PI3K) is a key player in the establishment of central sensitization, the spinal cord phenomenon associated with persistent afferent inputs and contributing to chronic pain states. We demonstrated electrophysiologically that PI3K is required for the full expression of spinal neuronal wind-up. In an inflammatory pain model, intrathecal administration of LY294002 [2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one], a potent PI3K inhibitor, dose-dependently inhibited pain-related behavior. This effect was correlated with a reduction of the phosphorylation of ERK (extracellular signal-regulated kinase) and CaMKII (calcium/calmodulin-dependent protein kinase II). In addition, we observed a significant decrease in the phosphorylation of the NMDA receptor subunit NR2B, decreased translocation to the plasma membrane of the GluR1 (glutamate receptor 1) AMPA receptor subunit in the spinal cord, and a reduction of evoked neuronal activity as measured using c-Fos immunohistochemistry. Our study suggests that PI3K is a major factor in the expression of central sensitization after noxious inflammatory stimuli. PMID- 18417707 TI - A novel embryonic nestin-expressing radial glia-like progenitor gives rise to zonally restricted olfactory and vomeronasal neurons. AB - Persistent neurogenesis is maintained throughout development and adulthood in the mouse olfactory epithelium (OE). Despite this, the identity and origin of different embryonic OE progenitors, their spatiotemporal induction and contribution to patterning during development, has yet to be delineated. Here, we show that the embryonic OE contains a novel nestin-expressing radial glia-like progenitor (RGLP) that is not found in adult OE, which is antigenically distinct from embryonic CNS radial glia. Nestin-cre-mediated lineage tracing with three different reporters reveals that only a subpopulation of nestin-expressing RGLPs activate "CNS-specific" nestin regulatory elements, and produce spatially restricted olfactory receptor neurons (ORNs) in zone 1 of the OE, and vomeronasal receptor neurons restricted to the VR1 zone. This dorsal-medial restriction of transgene-activating cells is also seen in the embryonic OE of Nestin-GFP transgenic mice, in which green fluorescent protein (GFP) is found in a subpopulation of GFP+Mash1+ neuronal progenitors, despite the fact that endogenous Nestin expression is found in RGLPs throughout the OE. Embryonic OE progenitors produce three biologically distinct colony subtypes in vitro, a subpopulation of which include nestin-expressing RGLPs during in vitro colony formation. When generated from Nestin-cre/ZEG mice, neurogenic colonies also produce GFP+Mash1+ progenitors and ORNs. We thus identify a novel neurogenic precursor, the RGLP of the OE and vomeronasal organ (VNO), and provide the first evidence for intrinsic differences in the origin and spatiotemporal potential of distinct progenitors during development of the OE and VNO. PMID- 18417708 TI - Dynamics of the microglial/amyloid interaction indicate a role in plaque maintenance. AB - Microglial cells aggregate around amyloid plaques in Alzheimer's disease, but, despite their therapeutic potential, various aspects of their reactive kinetics and role in plaque pathogenesis remain hypothetical. Through use of in vivo imaging and quantitative morphological measures in transgenic mice, we demonstrate that local resident microglia rapidly react to plaque formation by extending processes and subsequently migrating toward plaques, in which individual transformed microglia somata remain spatially stable for weeks. The number of plaque-associated microglia increased at a rate of almost three per plaque per month, independent of plaque volume. Larger plaques were surrounded by larger microglia, and a subset of plaques changed in size over time, with an increase or decrease related to the volume of associated microglia. Far from adopting a more static role, plaque-associated microglia retained rapid process and membrane movement at the plaque/glia interface. Microglia internalized systemically injected amyloid-binding dye at a much higher rate in the vicinity of plaques. These results indicate a role for microglia in plaque maintenance and provide a model with multiple targets for therapeutic intervention. PMID- 18417709 TI - Molecular mechanisms determining sperm motility initiation in two sparids (Sparus aurata and Lithognathus mormyrus). AB - Molecular mechanisms involved in sperm motility initiation in two sparids (Sparus aurata and Lithognathus mormyrus) have been studied. Our comparative study demonstrates that osmolality is the key signal in sperm motility activation in both species, whereas K(+) and Ca(2+) do not have any role. The straight-line velocity that resulted, however, was significantly different when measured in sperm activated with non-ionic and/or calcium-free solutions with respect to that measured in seawater-activated sperm. In both species, motility initiation depends on cAMP-dependent protein phosphorylation. The phosphorylation/dephosphorylation patterns that resulted in gilthead and striped sea bream were quite different. In gilthead sea bream, the phosphorylated proteins have molecular weights of 174, 147, 138, 70, and 9-15 kDa, whereas the dephosphorylated proteins have molecular weights of 76, 57, and 33 kDa. In striped sea bream, phosphorylation after sperm motility activation occurred on proteins of 174, 147, 103, 96, 61, 57, and 28 kDa, whereas only one protein of 70 kDa resulted from dephosphorylation. Matrix-assisted laser desorption ionization time of flight analyses allowed identification of the following proteins: In gilthead sea bream, the 9-15 kDa proteins that were phosphorylated after motility activation include an A-kinase anchor protein (AKAP), an acetyl-coenzyme A synthetase, and a protein phosphatase inhibitor, and in striped sea bream, 103- and 61-kDa proteins that were phosphorylated after motility activation were identified as a phosphatase (myotubularin-related protein 1) and a kinase (DYRK3), respectively. PMID- 18417710 TI - Identification of potential markers of oocyte competence expressed in bovine cumulus cells matured with follicle-stimulating hormone and/or phorbol myristate acetate in vitro. AB - Oocyte competence is the ability of the oocyte to complete maturation, undergo successful fertilization, and reach the blastocyst stage. Cumulus cells are indispensable for this process. Their removal significantly affects the blastocyst rates. Moreover, the properties and functions of cumulus cells are regulated by the oocyte. They also reflect the oocyte's degree of maturation. Our study was aimed at identifying markers of oocyte competence that are expressed in bovine cumulus cells. In a previous study in our laboratory, the blastocyst yield following FSH or phorbol myristate acetate (PMA) treatment was 45%%. Therefore, we tested four sets of conditions during the first 6 h of in vitro maturation (IVM): FSH (0.1 microg/ml), PMA (0.1 microM), FSH ++ PMA, and negative control. Extracts from each IVM treatment were hybridized against the same negative control on a microarray containing a partial library of differentially expressed transcripts in the cumulus of competent oocytes collected at 6 h after LH in vivo. Common positive clones between diffrentially treated cells were selected, and 15 candidates were validated by real-time PCR. Based on this, the main candidates expressed in cumulus cells and that could be valuable and indirect markers of oocyte competence are hyaluronan synthase 2 (HAS2), inhibin betaA (INHBA), epidermal growth factor receptor (EGFR), gremlin 1 (GREM1), betacellulin (BTC), CD44, tumor necrosis factor-induced protein 6 (TNFAIP6), and prostaglandin endoperoxide synthase 2 (PTGS2). These biomarkers could be potential candidates to predict oocyte competence and to select higher-quality embryos for transfer. Additionally, these indirect predictors of oocyte competence and follicular health could improve our knowledge of gene expression patterns in the cumulus and yield insights into the molecular pathways controlling oocyte competence. PMID- 18417711 TI - Integrin activation in bovine placentomes and in caruncular epithelial cells isolated from pregnant cows. AB - In the bovine synepitheliochorial placenta, restricted trophoblast invasion requires complex interactions of integrin receptors with proteins of the extracellular matrix (ECM) and integrin receptors of neighboring cells. Activated integrins assemble to focal adhesions and are linked to the actin cytoskeleton via signaling molecules including alpha-actinin (ACTN), focal adhesion kinase (PTK2 or FAK), phosphotyrosine, and talin (TLN1). Aims of this study were to assess integrin activation and focal adhesion assembly within epithelial cells of bovine placentomes and low-passage (not transformed) placentomal caruncular epithelial cells cultured on dishes coated with ECM proteins. Immunofluorescence analysis was performed to colocalize the signaling molecules ACTN, PTK2, phosphotyrosine, and TLN1 with each other and with beta(1)-integrin (ITGB1) in placentomal cryosections throughout pregnancy and in caruncular epithelial cells in vitro. Antibody specificity was confirmed by Western blot. Cells were cultured on uncoated dishes, and the dishes were coated with fibronectin (FN), laminin (LAMA), and collagen type IV (COL4), thereby statistically assessing cell number and qualitatively assessing the expression pattern of ITGB1, phosphotyrosine, and TLN1. Results demonstrated integrin activation and focal adhesion assembly in the placentome and that low-passage caruncular epithelial cells maintain integrin associated properties observed in vivo. Expression and/or colocalization of signaling molecules with ITGB1 confirmed, for the first time, integrin activation and participation in "outside-in" and "inside-out" signaling pathways. The prominent role of ECM, and FN in particular, in integrin signaling is supported by the in vitro enhancement of proliferation and focal adhesion expression. Thus, this in vitro model provides excellent potential for further mechanistic studies designed to elucidate feto-maternal interactions in the bovine placentome. PMID- 18417712 TI - MUC1 expression is increased during human placental development and suppresses trophoblast-like cell invasion in vitro. AB - Mucin (MUC)1 is a multifunctional mucin expressed by a variety of reproductive tract epithelia. Trophoblast invasion is essential for normal placental development. However, MUC1 expression in the human placenta throughout pregnancy and the role of MUC1 in trophoblast-like cell invasion are still unclear. In the present study, results from quantitative RT-PCR and Western blot demonstrated that MUC1 mRNA and MUC1 protein expression, respectively, increased with gestational age of the human placenta. Immunohistochemistry revealed that MUC1 in placental villi was mainly expressed by syncytiotrophoblasts throughout pregnancy and increased with gestational age. Interestingly, we found two populations of extravillous trophoblasts, MUC1-positive and MUC1-negative cells, in decidua. The numbers of MUC1-positive extravillous trophoblasts were increased during placental development. Furthermore, MUC1 overexpression significantly (P < 0.01) suppressed matrigel invasion of trophoblast-like JAR cells by 34.6% +/- 4.5% compared with control, which was associated with a decrease in MMP9 activity assessed by gelatin zymography. Our results suggest that MUC1 expression in the human placenta is increased during placental development, and its overexpression suppresses trophoblast-like cell invasion in vitro. PMID- 18417713 TI - Identification of novel bovine cumulus cell molecular markers predictive of oocyte competence: functional and diagnostic implications. AB - The present study was undertaken to discover molecular markers in bovine cumulus cells predictive of oocyte competence and to elucidate their functional significance. Differences in RNA transcript abundance in cumulus cells harvested from oocytes of adult versus prepubertal animals (a model of poor oocyte quality) were identified by microarray analysis. Four genes of interest encoding for the lysosomal cysteine proteinases cathepsins B, S, K, and Z and displaying greater transcript abundance in cumulus cells surrounding oocytes harvested from prepubertal animals were chosen for further investigation. Greater mRNA abundance for such genes in cumulus cells of prepubertal oocytes was confirmed by real-time RT-PCR. Elevated transcript abundance for cathepsins B, S, and Z also was observed in cumulus cells surrounding adult metaphase II oocytes that developed to the blastocyst stage at a low percentage following parthenogenetic activation versus those that developed at a high percentage. Functional significance of cumulus cell cathepsin expression to oocyte competence was confirmed by treatment of cumulus-oocyte complexes during in vitro oocyte maturation with a cell permeable cysteine proteinase (cathepsin) inhibitor. Inhibitor treatment decreased apoptotic nuclei in the cumulus layer and enhanced development of parthenogenetically activated and in vitro-fertilized adult oocytes to the blastocyst stage. Stimulatory effects of inhibitor treatment during meiotic maturation on subsequent embryonic development were not observed when oocytes were matured in the absence of cumulus cells. The present results support a functional role for cumulus cell cathepsins in compromised oocyte competence and suggest that cumulus cell cathepsin mRNA abundance may be predictive of oocyte quality. PMID- 18417714 TI - Expression patterns of SP1 and SP3 during mouse spermatogenesis: SP1 down regulation correlates with two successive promoter changes and translationally compromised transcripts. AB - Because of their prominent roles in regulation of gene expression, it is important to understand how levels of Krupple-like transcription factors SP1 and SP3 change in germ cells during spermatogenesis. Using immunological techniques, we found that both factors decreased sharply during meiosis. SP3 declined during the leptotene-to-pachytene transition, whereas SP1 fell somewhat later, as spermatocytes progressed beyond the early pachytene stage. SP3 reappeared for a period in round spermatids. For Sp1, the transition to the pachytene stage is accompanied by loss of the normal, 8.2-kb mRNA and appearance of a prevalent, 8.8 kb variant, which has not been well characterized. We have now shown that this pachytene-specific transcript contains a long, unspliced sequence from the first intron and that this sequence inhibits expression of a reporter, probably because of its many short open-reading frames. A second testis-specific Sp1 transcript in spermatids of 2.4 kb also has been reported previously. Like the 8.8-kb variant, it is compromised translationally. We have confirmed by Northern blotting that the 8.8-, 8.2-, and 2.4-kb variants account for the major testis Sp1 transcripts. Thus, the unexpected decline of SP1 protein in the face of continuing Sp1 transcription is explained, in large part, by poor translation of both novel testis transcripts. As part of this work, we also identified five additional, minor Sp1 cap sites by 5' rapid amplification of cDNA ends, including a trans spliced RNA originating from the Glcci1 gene. PMID- 18417715 TI - Induction of the acrosome reaction in black tiger shrimp (Penaeus monodon) requires sperm trypsin-like enzyme activity. AB - Trypsin-like enzymes in egg water (EW), a natural acrosome reaction (AR) inducer, are known for their importance in shrimp AR. In this report, we describe a unique phenomenon of the AR of black tiger shrimp (Penaeus monodon) sperm. It was completed within 45-60 sec and comprised only the acrosomal exocytosis and depolymerization of the sperm head anterior spike. We used peptidyl fluorogenic substrates to show the presence of trypsin-like enzymes in P. monodon EW and sperm, but minimal activities of chymotrypsin-like enzymes. In sperm, these trypsin-like enzymes existed both on the sperm surface and in the acrosome. The acrosomal enzyme was revealed as a 45-kDa band by fluorogenic substrate in-gel zymography. Although EW possessed high trypsin-like enzyme activities, they were not essential for the AR induction; EW pretreated with an irreversible trypsin inhibitor, or heat-inactivated EW (HI-EW), to abolish the trypsin-like activities could still induce the AR. The HI-EW-induced AR was inhibited by the presence of a membrane impermeant soybean trypsin inhibitor (SBTI) in the sperm suspension, indicating the significance of sperm-borne trypsin-like enzymes (on the surface and/or in the acrosome) in this AR process. However, pretreatment of sperm with SBTI followed by its removal from the suspension still allowed the AR to occur within 5 min of sperm exposure to HI-EW. Since trypsin-like activity of the SBTI pretreated sperm surface at 5 min after SBTI removal was at the minimal level, our results suggest the importance of the acrosomal trypsin-like enzyme in the AR process. PMID- 18417716 TI - Attenuation of retinal endothelial cell migration and capillary morphogenesis in the absence of bcl-2. AB - Apoptosis plays a critical role during development and in the maintenance of the vascular system. B-cell leukemia lymphoma 2 (bcl-2) protects endothelial cells (EC) from apoptosis in response to a variety of stimuli. Previous work from this laboratory demonstrated attenuation of postnatal retinal vascular development and retinal neovascularization during oxygen-induced ischemic retinopathy in bcl-2 deficient (bcl-2-/-) mice. To gain further insight into the function of bcl-2 in the endothelium, we isolated retinal EC from bcl-2+/+ and bcl-2-/- mice. Retinal EC lacking bcl-2 demonstrated reduced cell migration, tenascin-C expression, and adhesion to vitronectin and fibronectin. The bcl-2-/- retinal EC also failed to undergo capillary morphogenesis in Matrigel. In addition, using an ex vivo angiogenesis assay, we observed reduced sprouting from aortic rings grown in culture from bcl-2-/- mice compared with bcl-2+/+ mice. Furthermore, reexpression of bcl-2 was sufficient to restore migration and capillary morphogenesis defects observed in bcl-2-/- retinal EC. Mechanistically, bcl-2-/- cells expressed significantly less endothelial nitric oxide synthase, an important downstream effecter of proangiogenic signaling. This may be attributed to increased oxidative stress in the absence of bcl-2. In fact, incubation of retinal EC or aortic rings from bcl-2-/- mice with the antioxidant N-acetylcysteine rescued their capillary morphogenesis and sprouting defects. Thus, bcl-2-mediated cellular functions play important roles not only in survival but also in proangiogenic phenotype of EC with a significant impact on vascular development and angiogenesis. PMID- 18417717 TI - Volume-sensitive NADPH oxidase activity and taurine efflux in NIH3T3 mouse fibroblasts. AB - Reactive oxygen species (ROS) are produced in NIH3T3 fibroblasts during hypotonic stress, and H(2)O(2) potentiates the concomitant release of the organic osmolyte taurine (Lambert IH. J Membr Biol 192: 19-32, 2003). The increase in ROS production [5-(and-6)-carboxy-2', 7'-dichlorodihydrofluorescein diacetate fluorescence] is detectable after a reduction in the extracellular osmolarity from 335 mosM (isotonic) to 300 mosM and reaches a maximal value after a reduction to 260 mosM. The swelling-induced ROS production is reduced by the flavoprotein inhibitor diphenylene iodonium chloride (25 microM) but is unaffected by the nitric oxide synthase inhibitor N omega-nitro-l-arginine methyl ester, indicating that the volume-sensitive ROS production is NADPH oxidase dependent. NIH3T3 cells express the NADPH oxidase components: p22 phox, a NOX4 isotype; p47 phox; and p67 phox (real-time PCR). Exposure to the Ca2+-mobilizing agonist ATP (10 microM) potentiates the release of taurine but has no effect on ROS production under hypotonic conditions. On the other hand, addition of the protein kinase C (PKC) activator phorbol 12-myristate 13-acetate (PMA, 100 nM) or the lipid messenger lysophosphatidic acid (LPA, 10 nM) potentiates the swelling induced taurine release as well as the ROS production. Overexpression of Rac1 or p47 phox or p47 phox knockdown [small interfering (si)RNA] had no effect on the swelling-induced ROS production or taurine release. NOX4 knockdown (siRNA) impairs the increase in the ROS production and the concomitant taurine release following osmotic exposure. It is suggested that a NOX4 isotype plus p22 phox account for the swelling-induced increase in the ROS production in NIH3T3 cells and that the oxidase activity is potentiated by PKC and LPA but not by Ca2+. PMID- 18417718 TI - Octylphenol stimulates resistin gene expression in 3T3-L1 adipocytes via the estrogen receptor and extracellular signal-regulated kinase pathways. AB - Resistin is known as an adipocyte-specific secretory hormone that can cause insulin resistance and decrease adipocyte differentiation. It can be regulated by sexual hormones. Whether environmental estrogens regulate the production of resistin is still not clear. Using 3T3-L1 adipocytes, we found that octylphenol upregulated resistin mRNA expression in dose- and time-dependent manners. The concentration of octylphenol that increased resistin mRNA levels by 50% was approximately 100 nM within 6 h of treatment. The basal half-life of resistin mRNA induced by actinomycin D was lengthened by octylphenol treatment, suggesting that octylphenol decreases the rate of resistin mRNA degradation. In addition, octylphenol stimulated resistin protein expression and release. The basal half life of resistin protein induced by cycloheximide was lengthened by octylphenol treatment, suggesting that octylphenol decreases the rate of resistin protein degradation. While octylphenol was shown to increase activities of the estrogen receptor (ER) and MEK1, signaling was demonstrated to be blocked by pretreatment with either ICI-182780 (an ERalpha antagonist) or U-0126 (a MEK1 inhibitor), in which both inhibitors prevented octylphenol-stimulated phosphorylation of ERK. These results imply that ERalpha and ERK are necessary for the octylphenol stimulation of resistin mRNA expression. Moreover, U-0126 antagonized the octylphenol-increased resistin protein expression and release. These data suggest that the way octylphenol signaling increases resistin protein levels is similar to that by which it increases resistin mRNA levels; it is likely mediated through an ERK-dependent pathway. In vivo, octylphenol increased adipose resistin mRNA expression and serum resistin and glucose levels, supporting its in vitro effect. PMID- 18417719 TI - Endotoxin mediates recruitment of RNA polymerase II to target genes in acute renal failure. AB - Acute renal failure (ARF) sensitizes the kidney to endotoxin (LPS)-driven production of cytokines and chemokines. This study assessed whether this LPS hyperresponsiveness exists at the genomic level. Three heterogeneous mouse models of ARF were studied: Maleate nephrotoxicity, unilateral ureteral obstruction, and LPS preconditioning. In all cases, LPS was injected approximately 18 h after injury was induced, and over the next 0 to 90 min, RNA polymerase II recruitment to the genome at three LPS-responsive genes (TNF-alpha, monocyte chemoattractant 1 [MCP-1], and heme oxygenase-1 [HO-1]) was assessed by chromatin immunoprecipitation. LPS hyperresponsiveness was noted in each model, measured by exaggerated increases in TNF-alpha and MCP-1 mRNA (approximately two to 10 times higher than LPS-injected controls). Corresponding increases in the recruitment of RNA polymerase II to the TNF-alpha and MCP-1 genes were observed, and increased trimethylation of histone 3 lysine 4 (H3K4m3) at these sites may have played a role in this recruitment. Conversely, recruitment of RNA polymerase II to the HO 1 gene was suppressed ("tolerance"), and no increase in H3K4m3 was observed at HO 1 exons. The ARF-induced changes in mRNA did not correlate with mRNA stability, suggesting the mechanistic importance of RNA polymerase II-mediated transcriptional events. In conclusion, LPS hyperresponsiveness after ARF is likely mediated at the genomic level, possibly by H3K4m3. PMID- 18417720 TI - High perfusion pressure accelerates renal injury in salt-sensitive hypertension. AB - Renal injury in the Dahl salt-sensitive rat mimics human salt-sensitive forms of hypertension that are particularly prevalent in black individuals, but the mechanisms that lead to the development of this injury are incompletely understood. We studied the impact of renal perfusion pressure (RPP) on the development of renal injury in this model. During the development of salt-induced hypertension over 2 wk, the RPP to the left kidney was maintained at control levels (125 +/- 2 mmHg) by continuous servocontrol inflation of an aortic balloon implanted between the renal arteries; during the same period, the RPP to the right kidney rose to 164 +/- 8 mmHg. After 2 wk of a 4% salt diet, DNA microarray and real-time PCR identified genes related to fibrosis and epithelial-to mesenchymal transition in the kidneys exposed to hypertension. The increased RPP to the right kidney accounted for differences in renal injury between the two kidneys, measured by percentage of injured cortical and juxtamedullary glomeruli, quantified proteinaceous casts, number of ED-1-positive cells per glomerular tuft area, and interstitial fibrosis. Interlobular arteriolar injury was not increased in the kidney exposed to elevated pressure but was reduced in the control kidney. We conclude that elevations of RPP contribute significantly to the fibrosis and epithelial-to-mesenchymal transition found in the early phases of hypertension in the salt-sensitive rat. PMID- 18417721 TI - Downregulation of hepatic acetylation of drugs in chronic renal failure. PMID- 18417722 TI - The mechanism of phosphorus as a cardiovascular risk factor in CKD. AB - Hyperphosphatemia and vascular calcification have emerged as cardiovascular risk factors among those with chronic kidney disease. This study examined the mechanism by which phosphorous stimulates vascular calcification, as well as how controlling hyperphosphatemia affects established calcification. In primary cultures of vascular smooth muscle cells derived from atherosclerotic human aortas, activation of osteoblastic events, including increased expression of bone morphogenetic protein 2 (BMP-2) and the transcription factor RUNX2, which normally play roles in skeletal morphogenesis, was observed. These changes, however, did not lead to matrix mineralization until the phosphorus concentration of the media was increased; phosphorus stimulated expression of osterix, a second critical osteoblast transcription factor. Knockdown of osterix with small interference RNA (siRNA) or antagonism of BMP-2 with noggin prevented matrix mineralization in vitro. Similarly, vascular BMP-2 and RUNX2 were upregulated in atherosclerotic mice, but significant mineralization occurred only after the induction of renal dysfunction, which led to hyperphosphatemia and increased aortic expression of osterix. Administration of oral phosphate binders or intraperitoneal BMP-7 decreased expression of osterix and aortic mineralization. It is concluded that, in chronic kidney disease, hyperphosphatemia stimulates an osteoblastic transcriptional program in the vasculature, which is mediated by osterix activation in cells of the vascular tunica media and neointima. PMID- 18417723 TI - The subcellular localization of TRPP2 modulates its function. AB - TRPP2, also known as polycystin-2, is a calcium permeable nonselective cation channel that is mutated in autosomal dominant polycystic kidney disease but has also been implicated in the regulation of cardiac development, renal tubular differentiation, and left-to-right (L-R) axis determination. For obtaining further insight into how TRPP2 exerts tissue-specific functions, this study took advantage of PACS-dependent trafficking of TRPP2 in zebrafish larvae. PACS proteins recognize an acidic cluster within the carboxy-terminal domain of TRPP2 that undergoes phosphorylation and mediate retrieval of TRPP2 to the Golgi and endoplasmic reticulum (ER). The interaction of human TRPP2 with PACS proteins can be inhibited by a Ser812Ala mutation (TRPP2(S812A)), thereby allowing TRPP2 to reach other subcellular compartments, and enhanced by a Ser812Asp mutation (TRPP2(S812D)), thereby trapping TRPP2 in the ER. It was found that the TRPP2(S812A) mutant rescued cyst formation of TRPP2-deficient zebrafish larvae to the same degree as wild-type TRPP2, whereas the TRPP2(S812D) mutant was significantly more effective in normalizing the distorted body axis of TRPP2 deficient fish. Surprisingly, the TRPP2(S812D) mutant rescued the abnormalities of L-R asymmetry more effectively than either wild-type or TRPP2(S812A), suggesting that the ER localization of TRPP2 plays an important role in the development of normal L-R asymmetry. Taken together, these findings support the hypothesis that TRPP2 assumes distinct subcellular localizations to exert tissue specific functions. PMID- 18417724 TI - Circulating alloreactive T cells correlate with graft function in longstanding renal transplant recipients. AB - Monitoring for alloreactive memory T cells after organ transplantation may allow individualization of immunosuppression. Two pathways of T cell allorecognition have been implicated in chronic graft dysfunction: Direct (recipient T cells respond to donor peptides presented by donor antigen-presenting cells) and indirect (donor peptides are processed and presented by recipient antigen presenting cells). Previous studies have assessed these alloresponses only during the first 2 yr after kidney transplantation,so this study correlated the presence of circulating donor-reactive memory/effector T cells, primed by both pathways, in 34 longstanding living-donor renal transplant recipients using the highly sensitive IFN-gamma Elispot assay. Remarkably, 59% of patients had directly primed donor-reactive T cells, and their presence correlated directly with serum creatinine (P = 0.001) and inversely with estimated GFR (P = 0.042). Multivariate analysis revealed that hyporesponsiveness of direct, donor-specific T cells was the only variable that significantly correlated with graft function and that antidonor indirect alloreactivity was the only variable that significantly correlated with proteinuria. Interestingly, when both allorecognition pathways were considered together, patients with undetectable direct alloreactivity had better longterm graft function, independent of allosensitization by the indirect pathway. In conclusion, circulating donor-specific alloreactive T cells primed by both pathways are detectable long after transplantation and are associated with graft injury. Assessment of alloreactive memory/effector T cells might be helpful to tailor individual immunosuppression regimens for transplant recipients in the future. PMID- 18417726 TI - Introduction to the special issue on ADHD in adults: overcoming resistance while avoiding zealotry. PMID- 18417725 TI - Genetic-linkage mapping of complex hereditary disorders to a whole-genome molecular-interaction network. AB - Common hereditary neurodevelopmental disorders such as autism, bipolar disorder, and schizophrenia are most likely both genetically multifactorial and heterogeneous. Because of these characteristics traditional methods for genetic analysis fail when applied to such diseases. To address the problem we propose a novel probabilistic framework that combines the standard genetic linkage formalism with whole-genome molecular-interaction data to predict pathways or networks of interacting genes that contribute to common heritable disorders. We apply the model to three large genotype-phenotype data sets, identify a small number of significant candidate genes for autism (24), bipolar disorder (21), and schizophrenia (25), and predict a number of gene targets likely to be shared among the disorders. PMID- 18417727 TI - Case of a 67-year-old woman diagnosed with ADHD successfully treated with methylphenidate. PMID- 18417728 TI - Adult ADHD research: current status and future directions. PMID- 18417729 TI - Research forum on psychological treatment of adults with ADHD. AB - BACKGROUND: A literature search found five empirical studies of psychological treatment for adults with ADHD, out of 1,419 articles on ADHD in adults. Practice guidelines to date all recommend multimodal intervention, given that a significant number of patients cannot tolerate, do not respond to, or fail to reach optimal outcomes with medication alone. METHOD: This article provides a literature review and the recommendations of a forum of experts in the psychological treatment of adults with ADHD. RESULTS: Empirical studies of brief, structured, and short-term psychological interventions for adults with ADHD to date demonstrate moderate to large effect sizes. Methodological challenges include selection of control groups, broad-based measures of outcome, and the need for larger samples. CONCLUSION: Psychological treatment may play a critical role in the management of adults with ADHD who are motivated and developmentally ready to acquire new skills as symptoms remit. PMID- 18417730 TI - A push or a punch: distinguishing the severity of sibling violence. AB - Sibling violence is the most prevalent and least studied form of family violence, and little research has examined differences based on severity. This research examines more severe versus less severe forms of sibling violence. Using a subsample of married couples with two or more children ages 0 to 17 drawn from the 1976 National Survey of Physical Violence in American Families, the authors employ Conflict Tactics Scale items for child-to-child conflict to construct a measure of sibling violence severity. Drawing from several theoretical perspectives on family violence and peer aggression, the authors analyze the impact of macro-system variables, family stress and resources, and family subsystems on less severe and more severe sibling violence. Contextual factors are most important in explaining less severe sibling violence. Experience of parental violence and unpredictability are individual factors relevant to severe sibling violence. More research is needed to examine the etiology and impact of different forms of sibling violence. PMID- 18417731 TI - Why do rape survivors volunteer for face-to-face interviews? A meta-study of victims' reasons for and concerns about research participation. AB - There is growing interest in understanding how different research methods are perceived by victims of violence and what survivors will reveal to researchers (termed meta-research or meta-studies). The purpose of this project was to conduct a qualitative meta-study on why rape survivors chose to participate in community-based, face-to-face interviews. Participants mentioned four primary reasons for why they decided to participate in this study: (a) to help other survivors, (b) to help themselves, (c) to support research on rape/sexual assault, and (d) to receive financial compensation. Implications for designing research recruitment protocols are discussed. PMID- 18417732 TI - Health and occupational consequences of spouse abuse victimization among male U.S. Army soldiers. AB - Little is known about health and occupational outcomes of male spouse abuse victims. In all, 11,294 male spouse abuse victims with a history of spouse abuse perpetration, 3,277 victims without prior spouse abuse perpetration, and 72,855 nonvictims and nonperpetrators were followed for 12 years to assess army attrition and hospitalization risk. In multivariate Cox models controlling for age, race, education, rank, service time, and dependents, victims were at significantly greater risk for early army discharge and hospitalization than were nonvictims- particularly hospitalizations for depression, alcohol dependence, and mental health-even when the hospitalization occurred years after the abuse event. Victim-perpetrators had greater risk than did victim-nonperpetrators for both attrition (1.13, 95% confidence interval [CI] = 1.08-1.18; 1.05, 95% CI = 1.02 1.08, respectively) and hospitalizations (1.45, 95% CI = 1.38-1.52; 1.38, 95% CI = 1.27-1.49, respectively). College education was protective and deserves further inquiry. Male victims need greater support following spouse abuse. PMID- 18417733 TI - Curcumin structure-function, bioavailability, and efficacy in models of neuroinflammation and Alzheimer's disease. AB - Curcumin can reduce inflammation and neurodegeneration, but its chemical instability and metabolism raise concerns, including whether the more stable metabolite tetrahydrocurcumin (TC) may mediate efficacy. We examined the antioxidant, anti-inflammatory, or anti-amyloidogenic effects of dietary curcumin and TC, either administered chronically to aged Tg2576 APPsw mice or acutely to lipopolysaccharide (LPS)-injected wild-type mice. Despite dramatically higher drug plasma levels after TC compared with curcumin gavage, resulting brain levels of parent compounds were similar, correlating with reduction in LPS-stimulated inducible nitric-oxide synthase, nitrotyrosine, F2 isoprostanes, and carbonyls. In both the acute (LPS) and chronic inflammation (Tg2576), TC and curcumin similarly reduced interleukin-1beta. Despite these similarities, only curcumin was effective in reducing amyloid plaque burden, insoluble beta-amyloid peptide (Abeta), and carbonyls. TC had no impact on plaques or insoluble Abeta, but both reduced Tris-buffered saline-soluble Abeta and phospho-c-Jun NH(2)-terminal kinase (JNK). Curcumin but not TC prevented Abeta aggregation. The TC metabolite was detected in brain and plasma from mice chronically fed the parent compound. These data indicate that the dienone bridge present in curcumin, but not in TC, is necessary to reduce plaque deposition and protein oxidation in an Alzheimer's model. Nevertheless, TC did reduce neuroinflammation and soluble Abeta, effects that may be attributable to limiting JNK-mediated transcription. Because of its favorable safety profile and the involvement of misfolded proteins, oxidative damage, and inflammation in multiple chronic degenerative diseases, these data relating curcumin dosing to the blood and tissue levels required for efficacy should help translation efforts from multiple successful preclinical models. PMID- 18417734 TI - Pituitary homeobox 2 regulates adrenal4 binding protein/steroidogenic factor-1 gene transcription in the pituitary gonadotrope through interaction with the intronic enhancer. AB - Ad4BP/SF-1 [adrenal4 binding protein/steroidogenic factor-1 (NR5A1)] is a factor important for animal reproduction and endocrine regulation, and its expression is tightly regulated in the gonad, adrenal gland, ventromedial hypothalamic nucleus, and pituitary gonadotrope. Despite its functional significance in the pituitary, the mechanisms underlying pituitary-specific expression of the gene remain to be uncovered. In this study, we demonstrate by transgenic mouse assays that the pituitary gonadotrope-specific enhancer is localized within the sixth intron of the gene. Functionally, the enhancer recapitulates endogenous Ad4BP/SF-1 expression in the fetal Rathke's pouch to the adult pituitary gonadotrope. Structurally, the enhancer consists of several elements conserved among animal species. Mutational analyses confirmed the significance of these elements for the enhancer function. One of these elements was able to interact both in vitro and in vivo with Pitx2 (pituitary homeobox 2), demonstrating that pituitary homeobox 2 regulates Ad4BP/SF-1 gene transcription in the pituitary gonadotrope via interaction with the gonadotrope-specific enhancer. PMID- 18417736 TI - DAX-1 (dosage-sensitive sex reversal-adrenal hypoplasia congenita critical region on the X-chromosome, gene 1) selectively inhibits transactivation but not transrepression mediated by the glucocorticoid receptor in a LXXLL-dependent manner. AB - The glucocorticoid receptor (GR) mediates virtually all actions of glucocorticoids, and the nature and magnitude of a cell's response to these steroids are determined primarily by hormone concentration and GR signaling capacity. DAX-1 (dosagesensitive sex reversal-adrenal hypoplasia congenita critical region on the X-chromosome, gene 1) is an orphan nuclear receptor that functions as a corepressor, and deletion or mutation of DAX-1 causes a decrease in glucocorticoid production. However it is unclear whether DAX-1 also alters GR function as a transcription factor. Here, we demonstrate that DAX-1 acts as a novel selective GR modulator. It specifically inhibits ligand-dependent GR transactivation with little effect on GR-mediated transrepression. As demonstrated by coimmunoprecipitation and glutathione- S-transferase pull-down assays, DAX-1 physically interacts with GR, but this interaction does not influence either ligand-induced GR nuclear translocation or subsequent GR association with glucocorticoid-responsive elements. Instead, DAX-1 competes with coactivators such as GR-interacting protein 1 for binding to the receptor. Specifically, suppression of GR transactivation is mediated by the N-terminal half of DAX-1, and in particular the LXXLL motifs. Thus we demonstrate that DAX-1 directly modulates GR signaling in addition to affecting glucocorticoid hormone levels. PMID- 18417735 TI - Minireview: nuclear receptors and breast cancer. AB - Until recently, the study of nuclear receptor (NR) function in breast cancer biology has been largely limited to estrogen and progesterone receptors. The development of reliable gene expression arrays, real-time quantitative RT-PCR, and immunohistochemical techniques for studying NR superfamily members in primary human breast cancers has now revealed the presence and potential importance of several additional NRs in the biology of breast cancer. These include receptors for steroid hormones (including androgens and corticosteroids), fat-soluble vitamins A and D, fatty acids, and xenobiotic lipids derived from diet. It is now clear that after NR activation, both genomic and nongenomic NR pathways can coordinately activate growth factor signaling pathways. Advances in our understanding of both NR functional networks and epithelial cell growth factor signaling pathways have revealed a frequent interplay between NR and epithelial cell growth factor family signaling that is clinically relevant to breast cancer. Understanding how growth factor receptors and their downstream kinases are activated by NRs (and vice-versa) is a central goal for maximizing treatment opportunities in breast cancer. In addition to the estrogen receptor, it is predicted that modulating the activity of other NRs will soon provide novel prevention and treatment approaches for breast cancer patients. PMID- 18417737 TI - Guanine exchange factor RalGDS mediates exocytosis of Weibel-Palade bodies from endothelial cells. AB - The small GTP-binding protein Ral has been implicated in regulated exocytosis via its interaction with the mammalian exocyst complex. We have previously demonstrated that Ral is involved in exocytosis of Weibel-Palade bodies (WPBs). Little is known about intracellular signaling pathways that promote activation of Ral in response to ligand binding of G protein-coupled receptors. Here we show that RNAi-mediated knockdown of RalGDS, an exchange factor for Ral, results in inhibition of thrombin- and epinephrine-induced exocytosis of WPBs, while overexpression of RalGDS promotes exocytosis of WPBs. A RalGDS variant lacking its exchange domain behaves in a dominant negative manner by blocking release of WPBs. We also provide evidence that RalGDS binds calmodulin (CaM) via an amino terminal CaM-binding domain. RalGDS association to CaM is required for Ral activation because a cell-permeable peptide comprising this RalGDS CaM-binding domain inhibits Ral activation and WPB exocytosis. Together our findings suggest that RalGDS plays a vital role in the regulation of Ral-dependent WPB exocytosis after stimulation with Ca(2+)- or cAMP-raising agonists. PMID- 18417738 TI - Type I NKT cells protect (and type II NKT cells suppress) the host's innate antitumor immune response to a B-cell lymphoma. AB - Natural killer T (NKT) cells are a T-cell subpopulation known to possess immunoregulatory functions and recognize CD1d molecules. The majority of NKT cells express an invariant T-cell receptor (TCR) alpha chain rearrangement (Valpha14 Jalpha18 in mice; Valpha24 Jalpha18 in humans) and are called type I NKT cells; all other NKT cells are type II. In the current study, we have analyzed the roles for these NKT-cell subsets in the host's innate antitumor response against a murine B-cell lymphoma model in vivo. In tumor-bearing mice, we found that type I NKT cells conferred protection in a CD1d-dependent manner, whereas type II NKT cells exhibited inhibitory activity. Pro- and anti inflammatory cytokines secreted by splenocytes from tumor-bearing mice correlated with tumor progression. Myeloid cells (CD11b(+)Gr1(+)) were present in large numbers at the tumor site and in the spleen of tumor-bearing type I NKT-deficient mice, suggesting that antitumor immunosurveillance was inhibited by CD11b(+)Gr1(+) cells. Overall, these data suggest that there are distinct roles for NKT-cell subsets in response to a B-cell lymphoma in vivo, pointing to potential novel targets to be exploited in immunotherapeutic approaches against blood cancers. PMID- 18417739 TI - Phase I trial of tipifarnib in children with newly diagnosed intrinsic diffuse brainstem glioma. AB - The purpose of this study is to estimate the maximum-tolerated dose (MTD) and describe toxicities and preliminary clinical effects of tipifarnib, a farnesyltransferase (FTase) inhibitor, administered concurrently with radiation therapy in children with newly diagnosed intrinsic diffuse brainstem glioma (BSG). Children >or=3 and or =18 yr of age and had hemoglobin <11.0 g/dl and GFR 12 to 60 ml/min per 1.73 m(2). Patients received one of four epoetin alfa dosing regimens: 50 IU/kg three times per week, 10,000 IU once weekly, or 20,000 IU every 2 wk for 36 d or 40,000 IU every 4 wk for 64 d. Each regimen provided a similar dosage of epoetin alfa over 4 wk. Dosage adjustments were not permitted. RESULTS: Drug exposure to epoetin alfa over 4 wk, based on area under the curve, was somewhat higher with the extended interval regimens compared with the three-times-weekly regimen. Mean change in hemoglobin during the study period was similar for all regimens. No patients were transfused. Three patients experienced five serious adverse events, none of which was considered treatment related. CONCLUSIONS: Extended dosing interval regimens of epoetin alfa yielded modest pharmacokinetic differences but a similar pharmacodynamic response, suggesting that less frequent, higher dosages of epoetin alfa may be as effective as the current three-times-weekly regimen in anemic patients who have chronic kidney disease and are not on dialysis. PMID- 18417742 TI - Ascertainment and epidemiology of acute kidney injury varies with definition interpretation. AB - BACKGROUND AND OBJECTIVES: Differences in defining acute kidney injury (AKI) may impact incidence ascertainment. We assessed the effects of different AKI definition interpretation methods on epidemiology ascertainment. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: Two groups were studied at Texas Children's Hospital, Houston, Texas: 150 critically ill children (prospective) and 254 noncritically ill, hospitalized children receiving aminoglycosides (retrospective). SCr was collected for 14 d in the prospective study and 21 d in the retrospective study. Children with known baseline serum creatinine (bSCr) were classified by the pediatric Risk, Injury, Failure, Loss, End-Stage Kidney Disease (pRIFLE) AKI definition using SCr change (pRIFLE(DeltaSCr)), estimated creatinine clearance (eCCl) change (pRIFLE(DeltaCCl)), and the Acute Kidney Injury Network (AKIN) definition. In subjects without known bSCr, bSCR was estimated as eCCl = 100 (eCCl(100)) and 120 ml/min per 1.73 m(2) (eCCl(120)), admission SCr (AdmSCr) and lower/upper normative values (NormsMin, NormsMax). The differential impact of each AKI definition interpretation on incidence estimation and severity distribution was evaluated. RESULTS: pRIFLE(DeltaSCr) and AKIN led to identical AKI distributions. pRIFLE(DeltaCCl) resulted in 14.5% (critically ill) and 11% (noncritical) more patients diagnosed with AKI compared to other methods (P 0.05). Different bSCr estimates led to differences in AKI incidence, from 12% (AdmSCr) to 87.8% (NormsMin) (P 0.05) in the critically ill group and from 4.6% (eCCl(100)) to 43.1% (NormsMin) (P 0.05) in the noncritical group. CONCLUSIONS: AKI definition variation causes interstudy heterogeneity. AKI definition should be standardized so that results can be compared across studies. PMID- 18417743 TI - Cardiac evaluation before kidney transplantation: a practice patterns analysis in Medicare-insured dialysis patients. AB - BACKGROUND AND OBJECTIVES: Evaluation for ischemic heart disease (IHD) is a nonstandardized practice before kidney transplantation. We retrospectively studied pretransplant cardiac evaluation (CE) practices in a national sample of renal allograft recipients. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: The USRDS data for Medicare beneficiaries transplanted in 1991 to 2004 with Part A&B benefits from dialysis initiation through transplantation were examined. Clinical traits defining "high" expected IHD risk were defined as diabetes, prior IHD, or > or = 2 other coronary risk factors. Pretransplant CE were identified by billing claims for noninvasive stress tests and angiography. Patients were quantified with claims for coronary revascularization procedures between CE and transplant. Post-transplant acute myocardial infarction (AMI) events were abstracted from claims and death records. RESULTS: Among 27,786 eligible patients, 46.3% underwent CE before transplantation. Overall, 9.5% who received CE also received pretransplant revascularization, but only 0.3% of lower-risk patients undergoing CE had revascularization. The adjusted odds of transplant without CE increased sharply with younger age and shorter dialysis duration. Increased likelihood of transplant without CE also correlated with black race, female sex, and certain geographic regions. Among patients transplanted without CE, 3-yr incidence of post-transplant AMI was 3% in lower-risk and 10% in high-risk groups, and varied by individual traits within these groups. Among lower-risk patients transplanted without CE, blacks were higher risk for AMI than whites (adjusted hazards ratio 1.47, 95% CI 1.11-1.93). CONCLUSIONS: Observed practices demonstrate infrequent use of pretransplant revascularization after CE but also raise concern for socio demographic barriers to evaluation access. PMID- 18417744 TI - Greater epoetin alfa responsiveness is associated with improved survival in hemodialysis patients. AB - BACKGROUND AND OBJECTIVES: Among hemodialysis patients, achieved hemoglobin is associated with Epoetin alfa dose and erythropoietin responsiveness. A prospective erythropoietin responsiveness measure was developed and its association with mortality evaluated. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: Data from 321 participants were used and randomized to the hematocrit normalization arm of the Normal Hematocrit Cardiac Trial. Subjects were to receive a 50% Epoetin alfa dose increase at randomization. The prospective erythropoietin responsiveness measure was defined as the ratio of weekly hematocrit change (over the 3 wk after randomization) per Epoetin alfa dose increase (1000 IU/wk) corresponding to the mandated 50% dose increase at randomization. The distribution of responsiveness was divided into quartiles. Over a 1-yr follow-up, Cox proportional hazard modeling evaluated associations between this responsiveness measure and mortality. RESULTS: Erythropoietin responsiveness values ranged from -2.1% to 2.4% per week per 1000 IU. Although subjects were similar across response quartiles, mortality ranged between 14% and 34% among subjects in the highest and lowest response quartiles (P = 0.0004), respectively. After adjusting for baseline prognostic indicators, highest versus lowest responsiveness was associated with a hazard ratio of 0.41 (95% confidence interval, 0.20 to 0.87). CONCLUSION: Lower erythropoietin responsiveness is a strong, independent predictor of mortality risk and should be considered when evaluating associations between clinical outcomes and potential prognostic indicators, such as Epoetin alfa dose and achieved hemoglobin values. PMID- 18417745 TI - Identifying best practices in dialysis care: results of cognitive interviews and a national survey of dialysis providers. AB - BACKGROUND AND OBJECTIVES: Because there is wide variation in case-mix adjusted outcomes across dialysis facilities, it is possible that top-performing facilities use practices not shared by others. We sought to catalogue "best practices" that may account for interfacility variations in outcomes. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: This multidisciplinary study identified candidate best practices in dialysis through a staged process, including systematic review, cognitive interviews, and a national "virtual focus group" of dialysis providers. The resulting candidate practices were rank-ordered by perceived importance as determined by mean RAND Appropriateness Scores from a national survey of nephrologists, nurses, and opinion leaders. RESULTS: A total of 155 candidate best practices were identified. Among these, respondents believed dialysis outcomes are most strongly related to 1) characteristics of multidisciplinary care conferences, 2) technician proficiency in protecting vascular access, 3) training of nurses to provide education in fluid management, vascular access, and nutrition, 4) use of random and blinded audits of staff performance, and 5) communication and teamwork among staff. In contrast, there was wide disagreement about the importance of facility-based health maintenance practices, optimal staffing ratios, frequency of dialysis-based physician visits, and optimal frequency of multidisciplinary care. CONCLUSIONS: This study provides a "conceptual map" of candidate dialysis best practices and highlights areas of general agreement and disagreement. These findings can help the dialysis community think critically about what may define "best practice" and provide targets for future research in quality improvement. PMID- 18417746 TI - Novel links between the long pentraxin 3, endothelial dysfunction, and albuminuria in early and advanced chronic kidney disease. AB - BACKGROUND AND OBJECTIVES: Albuminuria and inflammation predict cardiovascular events. Pentraxin 3, an inflammatory mediator produced by, among others, endothelial cells, may have a role in atherogenesis. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: In 207 Swedish patients with stage 5 chronic kidney disease and 79 Turkish patients with type 2 diabetes and proteinuria and normal renal function, whether serum pentraxin 3 levels are associated with albuminuria and endothelial dysfunction was studied. RESULTS: Patients with stage 5 chronic kidney disease and a high degree of albuminuria more often had diabetes and higher levels of pentraxin 3, vascular cellular adhesion molecule-1, and blood pressure. Moreover, pentraxin 3 was independently associated with 24-h urinary albumin excretion. In patients with type 2 diabetes, pentraxin 3 was significantly higher than in control subjects. Patients with type 2 diabetes and more proteinuria had higher pentraxin 3, C-reactive protein, glycosylated hemoglobin, insulin, and homeostasis model assessment index as well as lower flow mediated dilation and serum albumin. Pentraxin 3 was positively correlated with C reactive protein, homeostasis model assessment index, and carotid intima-media thickness and negatively with flow-mediated dilation. Pentraxin 3 and glomerular filtration rate were independently associated with 24-h urinary protein excretion. Only pentraxin 3 and proteinuria were significantly and independently associated with flow-mediated dilation. CONCLUSIONS: In two different renal cohorts, one of stage 5 chronic kidney disease and one of type 2 diabetes and normal renal function, pentraxin 3 was independently associated with proteinuria. Moreover, both pentraxin 3 and proteinuria were associated with endothelial dysfunction in patients with type 2 diabetes. PMID- 18417747 TI - Calciphylaxis from nonuremic causes: a systematic review. AB - BACKGROUND AND OBJECTIVES: Calciphylaxis, or calcific uremic arteriolopathy, is a well-described entity in end-stage kidney disease and renal transplant patients; however, little systematic information is available on calciphylaxis from nonuremic causes. This systematic review was designed to characterize etiologies, clinical features, laboratory abnormalities, and prognosis of nonuremic calciphylaxis. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: A systematic review of literature for case reports and case series of nonuremic calciphylaxis was performed. Cases included met the operational definition of nonuremic calciphylaxis-histopathologic diagnosis of calciphylaxis in the absence of end stage kidney disease, renal transplantation, or acute kidney injury requiring renal replacement therapy. RESULTS: We found 36 cases (75% women, 63% Caucasian, aged 15 to 82 yr) of nonuremic calciphylaxis. Primary hyperparathyroidism, malignancy, alcoholic liver disease, and connective tissue disease were the most common reported causes. Preceding corticosteroid use was reported for 61% patients. Protein C and S deficiencies were seen in 11% of patients. Skin lesions were morphologically similar to calcific uremic arteriolopathy. Mortality rate was 52%, with sepsis being the leading cause of death. CONCLUSION: Calciphylaxis should be considered while evaluating skin lesions in patients with predisposing conditions even in the absence of end-stage kidney disease and renal transplantation. Nonuremic calciphylaxis is reported most often in white women. Mineral abnormalities that are invoked as potential causes in calcific uremic arteriolopathy are often absent, suggesting that heterogeneous mechanisms may contribute to its pathogenesis. Nonuremic calciphylaxis is associated with high mortality, and there is no known effective treatment. PMID- 18417748 TI - Performance of procedures by nephrologists and nephrology fellows at U.S. nephrology training programs. AB - BACKGROUND AND OBJECTIVES: Some procedures (e.g., placement of temporary hemodialysis catheters and kidney biopsies) are required in nephrology fellowship training. Others (e.g., placement of tunneled hemodialysis catheters, ultrasonography, and hemodialysis access interventions) are not required but are performed at some centers. To assess the procedures performed by nephrologists and nephrology fellows at U.S. adult nephrology training programs and the number of procedures required for fellow competency, a survey was conducted of all such training programs. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: An on-line survey was e-mailed to the directors of all U.S. adult nephrology fellowship programs in October to November 2007. RESULTS: Responses were received from 93 of 136 programs. Nephrologists and nephrology trainees perform native and transplant kidney biopsies in 98% to 99% of programs and, in about half of programs, also perform the ultrasound guidance. Diagnostic ultrasounds are performed at fewer programs. Temporary dialysis catheters are inserted at nearly all programs. Tunneled hemodialysis catheters and peritoneal dialysis catheters are placed at < or =20% of programs. Interventional procedures on hemodialysis access are performed at 13% to 21% of programs. Continuous renal replacement therapy is performed at 99% of programs, plasmapheresis at 40%. Many programs either do not specify a minimum number of supervised procedures that need to be performed to demonstrate competence or require a very limited number. CONCLUSIONS: Core procedures are performed at almost all programs. Experience and training in other procedures are variable. Many programs have limited requirements for the number of procedures trainees need to perform to demonstrate competence. PMID- 18417749 TI - Treating elderly people with diabetes and stages 3 and 4 chronic kidney disease. AB - Dedicated European and US clinical guidelines for type 2 diabetes in the elderly have been released, but they do not specifically address the issue of advanced chronic kidney disease (CKD) in older patients with diabetes. General clinical guidelines have been published on the treatment of patients with diabetic nephropathy (DN), but these address the issue of how to prevent progression and treat advanced DN without distinguishing between different age groups. Elderly patients with diabetes and stages 3 to 4 CKD have particular needs that differ from those of younger patients with the same conditions. This is mainly due to their frailty and shorter life expectancy. Differently tailored therapeutic strategies are needed, which may have less stringent targets; and the use of common drugs should be critically evaluated. The management agenda (metabolic control, low-protein diet, controlling BP, preventing progression of advanced DN, preventing cardiovascular outcomes) for these patients is discussed in light of the limits and perspectives of current guidelines. Intensive, simultaneous management of all items on the agenda may not be feasible for a proportion of older patients, and clinicians may have to give priority to reducing some risk factors rather than others, choosing between different therapies. PMID- 18417752 TI - Public discourse concerning swine genetics papers. PMID- 18417751 TI - Hyperinsulinemia in African-American adolescents compared with their American white peers despite similar insulin sensitivity: a reflection of upregulated beta cell function? AB - OBJECTIVE: African-American (AA) children are hyperinsulinemic and insulin resistant compared with American white (AW) children. Previously, we demonstrated that insulin secretion relative to insulin sensitivity was approximately 75% higher in AA compared with AW children, suggesting that hyperinsulinemia in AA children is not merely a compensatory response to lower insulin sensitivity. The aim of the present investigation was to assess whether glucose-stimulated insulin response is higher in AA versus AW adolescents who have comparable in vivo insulin sensitivity. RESEARCH DESIGN AND METHODS: The hyperinsulinemic-euglycemic and hyperglycemic clamp techniques were utilized to assess first- and second phase insulin secretion. Insulin secretion relative to insulin sensitivity was calculated as the glucose disposition index. RESULTS: AA adolescents compared with their AW peers with comparable insulin sensitivity and body composition had higher first-phase insulin concentrations. CONCLUSIONS: The quantitative relationship between insulin sensitivity and first-phase insulin appears to differ among AA and AW adolescents. PMID- 18417750 TI - Higher serum creatinine concentrations in black patients with chronic kidney disease: beyond nutritional status and body composition. AB - BACKGROUND AND OBJECTIVES: Serum creatinine concentrations tend to be higher in black than white individuals and people of other races or ethnicities. These differences have been assumed to be largely related to race-related differences in body composition, especially muscle mass. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: In a diverse population of hemodialysis patients, we compared mean serum creatinine concentrations in black versus nonblack patients, adjusting for case mix (age, gender, diabetes, and dialysis vintage), body size (height, weight), laboratory parameters of nutritional status (albumin, predialysis blood urea nitrogen, transferrin, phosphorus, glucose), dialysis dosage (urea reduction ratio), and parameters of bioelectrical impedance (resistance and reactance), proxies for body composition. RESULTS: Adjusted mean serum creatinine concentrations were significantly higher in black versus nonblack patients (11.7 versus 10.0 mg/dl; P < 0.0001). Black patients were roughly four-fold more likely to have a serum creatinine concentration >10 mg/dl and six-fold more likely to have a serum creatinine concentration >15 mg/dl. Higher serum creatinine concentrations were associated with a lower relative risk for death (0.93; 95% confidence interval 0.88 to 0.98 per mg/dl); the association was slightly more pronounced among nonblack patients. CONCLUSIONS: Serum creatinine concentrations are significantly higher in black compared with nonblack hemodialysis patients; these differences are not readily explained by differences in nutritional status or body composition. PMID- 18417753 TI - Dealing with charges of scientific misconduct. PMID- 18417754 TI - Swine genetics. PMID- 18417757 TI - Sexual tension in the airways: the puzzling duality of estrogen in asthma. PMID- 18417758 TI - Gas-phase ion chemistry of GeH(4)/B(2)H(6) mixtures. AB - The gas phase ion-molecule reactions in positively and negatively ionized germane/diborane mixtures have been studied by ion trap mass spectrometry. Reaction sequences and rate constants for the most interesting processes have been determined. In positive ionization, formation of Ge-B bonds exclusively occurs through condensation reactions of B(n)H(m)(+) ions with germane, followed by H(2) or BH(3) loss. No reactions of ions from germane with B(2)H(6) were observed under the experimental conditions used here. In negative ionization, the Ge(n)H(m)(-) (n = 1, 2) ion families react with diborane to yield the Ge(n)B(p)H(q)(-) (p = 1, 2) ions, again via dehydrogenation and BH(3) loss, while diborane anions proved to be unreactive. In both positive and negative ionization, Ge-B ions reach appreciable abundances. The present results afford fundamental information about the intrinsic reactivity of gas-phase ions and provide valuable indications about the first nucleation steps ultimately leading to amorphous Ge and B-doped semiconductor materials by chemical vapor deposition methods. PMID- 18417759 TI - Ion-molecule reactions involving methyl isocyanide and methyl cyanide. AB - Ion-molecule reactions involving methyl isocyanide and methyl cyanide have been performed in a new rf-only hexapole collision cell inserted in a large-scale tandem mass spectrometer. Beside protonation processes, N-methyl cyanogen ions (CH(3)N(+)CCN) and 1-methyleneiminium-1- ethylenium ions (CH(2)CN(+)CH(2)) have been produced in high yield and fully characterized by high-energy collisional activation. The unimolecular chemistry of the molecular ions of caffeine (1,3,7 trimethyl xanthine) has been revisited on the basis of these new results. PMID- 18417760 TI - Matrix-assisted laser desorption/ionization time-of-flight investigation of Nylon 6 and Nylon 66 thermo-oxidation products. AB - Matrix-assisted laser desorption/ionization mass spectrometry (MALDI) was used to determine the structure of the molecules produced in the thermo-oxidative degradation of Nylon 6 and Nylon 66, at 180 degrees C and 250 degrees C in air. The MALDI spectra of the thermo-oxidized nylons provide information on the structure and end groups of the oligomers produced in the oxidation process. Our results show that the thermo-oxidation of both Ny6 and Ny66 proceeds through a hydrogen abstraction and subsequent formation of hydroperoxide intermediates. The latter decompose, yielding oligomers containing aldehydes, amides and methyl terminal groups. The aldehydes undergo further oxidation to produce carboxylic end groups. The formation of cyclopentanone terminal groups is also observed in the case of Nylon 66. Oligomers with structures deriving from Norrish-type degradation processes were not detected here for either Ny6 or Ny66. PMID- 18417761 TI - Conformational analysis of dynorphin A (1-13) using hydrogen-deuterium exchange and tandem mass spectrometry. AB - Trifluoroethanol (TFE)-induced conformational changes in dynorphin A (1-13) were investigated using charge-state distribution (CSD) and hydrogen-deuterium exchange (HDX), combined with electrospray ionization (ESI) mass spectrometry (MS). Individual amino acids involved in secondary structural elements were identified by collision-induced dissociation-tandem mass spectrometry (MS/MS). It was observed that dynorphin A (1-13) largely exists in an unfolded conformation and a folded structure in increasing concentrations of TFE. In 50% TFE, it forms an alpha-helix that encompasses residues 1-9 and remains flexible from residues 10 to 13. PMID- 18417762 TI - Detection of dehydroepiandrosterone misuse by means of gas chromatography- combustion-isotope ratio mass spectrometry. AB - According to World Anti-Doping Agency (WADA) rules (WADA Technical Document TD2004EAAS) urine samples containing dehydroepiandrosterone (DHEA) concentrations greater than 100 ng ML(-1) shall be submitted to isotope ratio mass spectrometry (IRMS) analysis. The threshold concentration is based on the equivalent to the glucuronide, and the DHEA concentrations have to be adjusted for a specific gravity value of 1.020. In 2006, 11,012 doping control urine samples from national and international federations were analyzed in the Cologne doping control laboratory, 100 (0.9%) of them yielding concentrations of DHEA greater than 100 ng mL(-1). Sixty-eight percent of the specimens showed specific gravity values higher than 1.020, 52% originated from soccer players, 95% were taken in competition, 85% were male urines, 99% of the IRMS results did not indicate an application of testosterone or related prohormones. Only one urine sample was reported as an adverse analytical finding having 319 ng mL(-1) DHEA (screening result), more than 10,000 ng mL(-1) androsterone and depleted carbon isotope ratio values for the testosterone metabolites androsterone and etiocholanolone. Statistical evaluation showed significantly different DHEA concentrations between specimens taken in- and out-of- competition, whereas females showed smaller DHEA values than males for both types of control. Also a strong influence of the DHEA excretion on different sport disciplines was detectable. The highest DHEA values were detected for game sports (soccer, basketball, handball, ice hockey), followed by boxing and wrestling. In 2007, 6622 doping control urine samples were analyzed for 3alpha,5-cyclo-5alpha-androstan-6beta-ol-17-one (3alpha,5-cyclo), a DHEA metabolite which was described as a useful gas chromatography-mass spectrometry (GC-MS) screening marker for DHEA abuse. Nineteen urine specimens showed concentrations higher than the suggested threshold of 140 ng mL(-1), six urine samples yielded additionally DHEA concentrations higher than 100 ng mL(-1), none of them showing positive IRMS findings. These results should be taken into consideration in future discussions about threshold values for endogenous steroids in doping control. PMID- 18417763 TI - Letter: electron impact mass spectrometry study of a series of substituted 5- aminoalkylmethyl-cytosines and their 1-N-(o-, m- and p-)bromobenzyl-substituted derivatives. AB - The electron-impact mass spectra of 5-aminoalkylmethyl-substituted cytosine and of their 1-N- o-(m- or p-)bromobenzyl-substituted derivatives are discussed. The influence of 5-aminoalkyl and 1-N-bromobenzyl substituents on the mode of mass fragmentation occurring upon electron impact ionization was demonstrated. The fragmentation pathways of all compounds and the characteristic ions in these spectra originating from the McLafferty rearrangement, RDA reactions and simple cleavages are presented. PMID- 18417765 TI - History of development of nephrology. PMID- 18417764 TI - Letter: collision-induced dissociation of [metal(L)(2)](2+) complexes (metal = Cu, Ca and Mg) of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine allows distinction of the acyl groups at the sn1 and sn2 positions. AB - The collision induced dissociation (CID) spectra of the divalent metal complexes of 1-palmitoyl-2-oleoyl-sn- glycero-3-phosphocholine, [Metal(lI)(L)(2)](2+) (where metal = Cu(2+), Mg(2+) and Ca(2+), L = [16:0/18:1GPCho]), formed by electrospray ionization, reveal interesting metal dependant fragmentation chemistry. Six main classes of reaction are observed corresponding to: two competing carboxylate abstraction pathways (from the sn1 and sn2 positions); phosphate abstraction; competing losses of the two different carboxylic acids from the sn1 and sn2 positions; loss of a protonated ligand, [L + H](+). The relative ratios of the competing carboxylate abstraction reactions are dependant on the metal, with the Cu and Ca complexes favouring the abstraction of the larger carboxylate (18:1) and the Mg complex favoring the abstraction of the smaller carboxylate (16:0). PMID- 18417766 TI - Hospital acquired acute renal failure. AB - This study was performed to evaluate hospital-acquired renal insufficiency with special reference to the causative factors, clinical course and patient management. A total of 100 patients who were admitted to medical and surgical wards at the King Hussein Medical Center, Amman, Jordan over a 15-month period and who developed renal failure during their hospital stay, were referred to, and seen at the nephrology department. The patients ages varied from 18 to 77 years; there were 59 males and 41 females. Reduced renal perfusion, major surgery and administration of nephrotoxic drugs accounted for the majority of episodes of renal failure (77%). In general, non-oliguric patients fared better, they required dialysis less frequently and had lower mortality compared to the oliguric patients. Severity of renal failure and multi-organ involvement adversely affected the prognosis. However, age of the patient had no influence on the outcome. PMID- 18417767 TI - Prevalence of genital disorders in jordanian school boys. AB - A prospective field study was conducted to determine the prevalence rate of cryptorchidism, hypospadias and other genital disorders in school children in the eastern part of Jordan. A total of 2495 school boys from 10 different schools were examined in May-June, 1995. Standardized examination and classification criteria were used. Two general practitioners and two pediatricians carried out the study. The various genital disorders detected during the screening were as follows: Undescended testis, 1.96%; Inguinal hernia, 0.80%; Varicocele, 1.08%; Hydrocele, 0.52%; Hypospadias, 0.92%. The prevalence rates of genital disorders in this study were similar to those reported by others. Early recognition and treatment of genital disorders in school children is mandatory in order to prevent potentially adverse effects. PMID- 18417768 TI - Endoscopic suspension of bladder neck for treatment of female stress urinary incontinence. AB - Between the years 1984 and 1993, a total of 72 female patients underwent endoscopic suspension of bladder neck for treatment of stress urinary incontinence at the King Hussein Medical Center and Hamad General Hospital, Amman, Jordan. The age of the study patients ranged between 32 and 75 years (mean: 46.5 years). Fifteen (20.8%) had history of previous operations for incontinence. The overall success rate achieved was 93.1%. Post-operative complications were encountered in seven (9.8%) patients within follow-up period of 4 to 43 months. Treatment failure occurred in five patients (6.9%). Our experience further confirms that endoscopic suspension of the bladder neck is a simple and reliable procedure in the treatment of female stress urinary incontinence. PMID- 18417769 TI - Relationship of cyclosporine and gingival overgrowth. AB - Cyclosporine is used as an immunosuppressive agent in organ transplantation. It has several side effects including gingival overgrowth. We evaluated 30 renal transplant patients on cyclosporine for the degrees of the gingival overgrowth. All the patients were males with mean age of 37.5 years. Eighteen patients (60%) had moderate to severe degrees of gingival overgrowth. No correlation was found with the trough levels of cyclosporine in blood. We conclude that gingival overgrowth is associated with cyclosporine immunosuppressive therapy but may not be dose related. The effect of other factors such as dental plaque and oral hygiene should be further studied. PMID- 18417770 TI - Skin Cancer and HLA Association in Renal Transplant Recipients. AB - Mismatching for HLA antigens in renal transplant recipients effects the graft survival and may also be responsible for the development of malignancy including skin cancer due to the intensive immunosuppression, continuous antigenic stimulation or both. In this article, available data are reviewed and an attempt is made to show that some specific immunologic and immuno-genetic factors like the HLA system also play a role in the development of skin cancer in renal transplant recipients. PMID- 18417771 TI - An unusual cause of recurrent bacteriuria in a kidney transplant recipient. AB - A 17-year old Saudi male presented to the transplant clinic of the King Fahad National Guard Hospital, Riyadh two weeks after undergoing a living unrelated kidney transplant in India. Graft function was normal and routine urine culture grew Pseudomonas aeruginosa for which he was treated; a follow-up urine culture was negative. Five months later, routine urine culture again documented the presence of Pseudomonas while the patient continued to be asymptomatic with normal graft function. Abdominal ultrasound showed shrunken native kidneys and a normal graft. A slightly echogenic mass was detected in the bladder. Cystocopy showed a retained non-absorbable suture at the area of the uretero-vesical anastomosis. Cystoscopic removal of the suture was carried out following which urine cultures have remained persistently negative. PMID- 18417772 TI - Rifampicin-induced acute renal failure: a case report. AB - Acute oliguric renal failure (ARF) developed in a patient two weeks after he was started on intermittent anti-tuberculous therapy including rifampicin. The clinical picture was compatible with acute allergic interstitial nephritis. Renal histology revealed mainly acute tubular necrosis with mild tubulo-interstitial mononuclear cellular infiltrate. Intermittent therapy, as in our patient, has been the major factor in the development of rifampicin induced ARF in cases reviewed in the literature. PMID- 18417773 TI - Renal replacement therapy in pakistan. PMID- 18417775 TI - The saudi journal of kidney diseases and transplantation: the new look. PMID- 18417774 TI - Medical complications following live related renal transplantation: a single center experience. PMID- 18417776 TI - Pregnancy in women treated with dialysis. PMID- 18417777 TI - Acquired Cystic Renal Disease in Patients Receiving Long-term Hemodialysis. AB - Thirty seven patients with End Stage Renal Disease (ESRD) on maintenance hemodialysis at our center were studied for prevalence of Acquired Cystic Renal Disease (ACRD). Eighteen (49%) patients had documented ACRD. The mean age was 50.4 years. The proportion of patients with ACRD rose with increasing duration of dialysis. In our study, ACRD was found in 71.4% of patients who had dialysis for more than four years and in 43.3% of patients who underwent dialysis for less than four years. All of our patients with ACRD remained asymptomatic except one who had flank pain and macro-hematuria. There was no clinical or ultrasonographic evidence of renal neoplasm in any of these patients. PMID- 18417778 TI - The impact of renal biopsy in the clinical management of childhood renal disease. AB - This study was carried out to determine whether renal biopsy altered patient management or expected prognosis among children with renal disease. A retrospective analysis of case records of 106 children with renal disease having adequate renal biopsies was made. The diagnosis was altered in 47.2% and therapeutic approach was changed in 33% of the patients after obtaining the renal biopsy findings. Also, the estimate of prognosis was altered in 31.1% of the cases. These results were greater for patients with steroid resistant and congenital nephrotic syndrome, acute renal failure, asymptomatic urinary abnormalities and mixed nephritic-nephrotic syndrome. Subsequent management and prognosis was least affected by the biopsy results in patients with steroid sensitive nephrotic syndrome and chronic renal failure. PMID- 18417779 TI - Etiology of end-stage renal disease in two regions of saudi arabia. AB - A total of 127 patients with end-stage renal disease (ESRD) including 45 from Al Madinah and 82 from the Gizan regions of Saudi Arabia were studied to determine the etiology of ESRD. The categorization into various diagnoses were according to established criteria. A total of 22 renal biopsies were also obtained to aid in the diagnosis. The mean age of the study patients was 50 years in Al-Madinah region and 37 years in Gizan region. Overall, the etiology could be established in 56 patients (44.1%) including 31 patients (68.9%) in Al-Madinah and 25 patients (30.5%) in Gizan regions. Diabetic nephropathy was the commonest cause of ESRD in Al-Madinah (45.2%) while obstructive nephropathy was the commonest identifiable cause in Gizan (24%). Our study suggests that diabetes and obstruction are the main conditions against which strategies for prevention and treatment should be directed. PMID- 18417780 TI - Management of the brain-dead organ donor. PMID- 18417781 TI - Pregnancy in hemodialysis patients. AB - Pregnancy is an infrequent event in women at child bearing age who are on chronic dialysis therapy. To evaluate the incidence and the various therapeutic interventions on the outcome of pregnancy we studied 80 women on chronic hemodialysis. There were six pregnancies in five patients over six years of follow up (1988 -1994). To reduce the risk factors associated with pregnancy, we tried to control the blood pressure to a level below l50/90mmHg, to increase the duration and frequency of hemodialysis, to keep the hemoglobin level in the blood above 80 gm/L and to minimize the risk of bleeding by the usage of regional heparinization during the dialysis sessions. Also we kept predialysis BUN below 25mmol/L and used bicarbonate based dialysate. Two pregnancies ended with spontaneous abortion, three with premature delivery and only one had delivery on time. Two of the infants born alive died later during the neonatal period, while the other two are still alive - one, a girl, is now 5 years old and the other, a boy, is now 1 (1/2) years old. None of our pregnant women died. We conclude that pregnancy is of low incidence in hemodialysis patients, and is mostly unsuccessful. However, the modern advances in hemodialysis therapy, the wide choice of anti-hypertensive, the correction of anemia, and the improvement in the general health of HD patients may improve the outcome. PMID- 18417782 TI - Sertoli Cell Tumor of Testes in a Child with Peutz-Jeghers syndrome. AB - The Peutz-Jeghers syndrome (PJS) is known to be commonly associated with ovarian tumors. However, its association with testicular tumors is uncommon. We report an eight-year old boy who presented with gynecomastia and had Sertoli cell tumor of both testes with high aromatase activity. To the best of our knowledge, this is the seventh such case being reported in the literature and the presenting manifestation in all seven was gynecomastia. PMID- 18417783 TI - Nephrocalcinosis and Bartter's Syndrome. AB - We report a case of a child with symptoms and signs of Bartter's syndrome and nephrocalcinosis. This report discusses the manifestations and management of this rare disease at a very early age. PMID- 18417784 TI - Vesicoureteral reflux in children. PMID- 18417785 TI - Recurrent urinary tract infection in a renal transplant recipient. PMID- 18417786 TI - Acquired cystic renal disease. PMID- 18417787 TI - Recent developments in vascular access for hemodialysis. AB - Vascular access surgery is a permanent challenge for the vascular surgeon and is a very dynamic, inter-disciplinary field in modern medicine. High technical skill, extreme care and continuity to gain as much experience as possible are required. Priority should be given to the patients' native vessels. Nevertheless, grafts do have a place provided special attention is paid to the restricted indications. Close co-operation of the vascular surgeon, the nephrologist and the radiologist is essential. The nephrologist is responsible for access surveillance and for early detection of fistula dysfunction. Dialysis related parameters should be observed routinely; clinical examination and if necessary ultrasonographic and angiographic technique will give further information. Recently introduced methods such as permanent central venous catheters and an increasing variety of interventional procedures must be critically evaluated in the future. PMID- 18417788 TI - Central vein stenosis in patients with prior subclavian vein catheterization for maintenance dialysis. AB - We evaluated the prevalence of central vein stenosis in patients with prior central vein catheterization for vascular access for hemodialysis. A total of 36 patients with end-stage renal disease (ESRD) who were referred to the division of vascular surgery at King Khalid University Hospital in Riyadh were evaluated. Bilateral ascending venogram was performed in all cases. The patients were divided into three groups: Group I included 38 extremities in 32 patients who were referred for construction of an arteriovenous fistula. All these patients had previous history of subclavian vein catheterization. This included six patients who had bilateral catheter insertion. Venography demonstrated greater than 50% central vein lesions in 13 of the 38 extremities yielding a prevalence of 34%. Group II included four extremities in four patients who were on hemodialysis for many years all of whom presented with features suggestive of venous obstruction in their already existing arteriovenous fistulae. All four patients gave history of prior subclavian vein catheterization for dialysis. Three of these patients had > 50% subclavian vein stenosis while one showed complete occlusion of the vein. Group III included 30 extremities without history of prior central vein cannulation and none of them showed any lesions on venography. Our study further confirms previous reports that show a high prevalence of central vein stenosis following use of subclavian catheter as vascular access. We suggest that the subclavian route should be abandoned in patients with ESRD and temporary venous cannulation, if needed, should be performed in the femoral or internal jugular veins. Also, in individuals with history of prior central vein cannulation, venography is strongly recommended before an arteriovenous fistula is made. PMID- 18417789 TI - Wilm's Tumor in Children: A 10-year Experience from the Eastern Province of Saudi Arabia. AB - Eighteen children with biopsy-proven Wilm's tumor were diagnosed and treated between 1985-1994 at the King Fahd Hospital of the University, Alkhobar, Saudi Arabia. There were eight males and 10 females. The median age at diagnosis was 2.5 years. Two patients (11%) had stage I, six patients (33%) stage II, eight patients (44%) stage III and one patient each (6%) had stages IV and V respectively. Seventeen patients (94%) had favorable histology and one (6%) had anaplastic type. One patient had bilateral tumor and one had associated aniridia. Follow-up was from 2 to 10 years (mean:5 years). The treatment administered was according to the recommendations of the National Wilm's Tumor Study II and III. Two patients died resulting in an overall disease-free survival of 88.8%. Our study further confirms that Wilm's tumor is a curable malignancy even in advanced stage. PMID- 18417790 TI - CAPD: Experience at Al Madinah Al Munawarah. AB - CAPD is not as popular mode of renal replacement therapy (RRT) in this country as it is in the west and hence the experience with CAPD is limited. At our center 16 patients (8 Males, 8 Females) with ESRD with a mean age of 46.3 +/- 15.6 years were managed by CAPD during 1990-1994. CAPD was the first option as a renal replacement therapy (RRT) in seven (44%) and second option in nine (56%) patients. Indications for CAPD were: Vascular access problems in 10 (62.5%), cardiovascular instability in three (19%), patient preference in two (12.5%) and non-availability of nearby HD center in one (6%) patient. Standard safe leur-lock system was used for dialysis. All patients were well dialyzed with good quality of life. Peritonitis was the main complication encountered (one episode/eight patient months), cause of hospital admission and reason for returning back to hemodialysis (HD). Other complications encountered were intraperitoneal bleeding in three, accidental cutting of outline in two, leakage in one and exit site infection in one. Average survival of this method was 15 + 10.5 months (range 1 37 months). At the time of reporting this study seven patients continued to be on CAPD, six were transferred back to hemodialysis, one was transplanted and two died due to other reasons. We conclude that CAPD is an acceptable mode of RRT in this part of the world. PMID- 18417791 TI - Rhabdomyolysis and acute renal failure secondary to toxic material abuse in morocco. AB - We have studied 26 patients with rhabdomyolysis complicated with acute renal failure. The causes of rhabdomyolysis were dominated by paraphenylene diamine intoxication (PPD) (13 cases). The other etiologies were: crush syndrome (6 cases), eclampsia (3 cases), intense physical effort (1 case), butane intoxication (1 case), postural rhabdomyolysis induced by a prolonged surgical operation for pheochromocytoma (1 case). None of the patients had any past medical history of muscle or renal disease. Serum level of creatinine phosphokinase varied from 1200 to 5400 IU/L. The mean peak of serum creatinine level was 620 umol/1. Therapy included forced alkaline diuresis in all cases. Hemodialysis was required in 18 cases. All but eight patients survived. We conclude that rahabdomyolysis could be caused by intoxication, which was a prime cause in this study. Prevention may be possible with restriction of use of toxic products. PMID- 18417792 TI - Hepatitis C Virus Infection among Hemodialysis Patients in the Najran Region of Saudi Arabia. AB - Forty seven patients with end-stage renal disease (ESRD) on maintenance hemodialysis at the Najran General Hospital, Najran, Saudi Arabia were studied for prevalence of hepatitis C virus (HCV) infection. A total of 24 patients (51.1%) were found to be positive for anti-HCV antibodies. Duration of hemodialysis was longer (p = < 0.025) in anti-HCV positive patients (4.9 +/- 2.9 years) as compared to anti-HCV negative subjects (1.7 + 1.4 years). No significant relationship was found between blood transfusions and anti-HCV positivity. Ten (41.7%) of the anti-HCV positive patients showed elevated liver enzymes in comparison with two (8.7%) of the seronegative patients. This difference was statistically significant (p = < 0.05). Liver biopsies from eight of the anti-HCV positive patients demonstrated cirrhosis in one, chronic active hepatitis in three, chronic persistent hepatitis in three and normal histology in one patient. There was no correlation between liver enzymes and the severity of histopathological findings on liver biopsy. PMID- 18417793 TI - Consensus and Controversies on HLA Matching and Crossmatching in Transplantation. AB - Transplantation of organs is becoming increasingly successful. What was once an experimental and life-saving emergency procedure has now been transformed into a life-enhancing and technologically advanced form of therapy. Histocompatibility testing between the donor and recipient consist of identification of HLA antigens known as tissue typing, and testing for pre-formed antibodies in the recipient against the donor HLA antigens, known as crossmatching. The importance of matching of HLA antigens has been debated ever since the first successful renal transplant was performed in 1954. Powerful and newer immunosuppressive drugs have not been able to establish an operational state of long-term organ tolerance. In this article, an attempt is made to show the clear and obvious effect of HLA antigen matching on the long-term graft survival. Also, the merits and demerits of different crossmatching methods and their proper interpretation and application to improve the overall graft survival are discussed. PMID- 18417795 TI - Renal replacement therapy in egypt. PMID- 18417794 TI - Association of multicystic dysplasia and crossed nonfused renal ectopia: a case report. AB - Crossed renal ectopia is an unusual anomaly, 90% of which is fused to the ipsilateral kidney. Multicystic dysplasia in association with crossed non-fused renal ectopia is a very rare phenomenon and can produce a palpable abdominal mass. We report a 2-year old girl who presented with a mass in the abdomen. Detailed evaluation including laparotomy confirmed a diagnosis of multicystic dysplasia in a crossed non-fused ectopic kidney. This diagnostic entity should be considered, when a mass is found in the abdomen or pelvis, and when no kidney can be found in it's normal ipsilateral position. PMID- 18417796 TI - Hospital acquired renal failure. PMID- 18417798 TI - An appreciation. PMID- 18417797 TI - History of development of nephrology. PMID- 18417799 TI - The special issue on pediatric nephrology: a trailer. PMID- 18417800 TI - Hereditary renal diseases. PMID- 18417802 TI - Hereditary and acquired renal tubular disorders: the saudi experience. PMID- 18417801 TI - Renal tubular acidosis in children. PMID- 18417803 TI - Acute renal failure in the neonate. PMID- 18417804 TI - Chronic renal failure in children: medical management. PMID- 18417805 TI - Growth in children with chronic renal disease. PMID- 18417806 TI - Continuous peritoneal dialysis in children: past, present and future. AB - Continuous peritoneal dialysis (CPD) in children has been an important mode of renal replacement therapy (RRT) since 1978. Continuous ambulatory peritoneal dialysis (CAPD) was used initially in children and then, in infants. Automated peritoneal dialysis (APD) was introduced in the eighties. Currently. CPD is the predominant mode of RRT in children, and 3/4 of these patients are managed with APD. Certain events have evolved with time, such as: a) growth and nutrition; b) renal osteodystrophy; c) anemia; and d) training. Complications from CPD continue to be a challenge and include: a) peritonitis; b) hernias and c) stress. Future directions in pediatric CAPD are: a) applying techniques to measure dialysis adequacy; b) improvement in dialysis solutions, such as pH, osmolality and different osmotic agents; c) additives to the dialysis fluid, such as hormones, growth factors; d) improvements in catheters and cyclers; and e) strategies to reduce peritonitis rates. There are challenges for CPD in the future; however, it appears to be the most practical and cost-effective dialysis therapy for children with end-stage renal disease. PMID- 18417807 TI - Spectrum of glomerular disease among children in saudi arabia. AB - A total of 376 renal biopsies performed in several hospitals in Riyadh, on children with glomerulonephritis were reviewed. Focal segmental glomerulosclerosis (FSGS) was the most common glomerulopathy found and accounted for 120 cases (31.9%), followed by mesangial proliferative glomerulonephritis in 99 (26.3%). Minimal change nephrotic syndrome was seen in 55 (14.6%), membranoproliferative glomerulonephritis in 30 (8.0%), membranous glomerulonephritis in 18 (4.8%), IgA nephropathy in 15 (4.0%), post-infectious glomerulonephritis in 15 (4.0%), Alport syndrome in 14 (3.7%) and rapidly progressive glomerulonephritis in 10 (2.7%). Our findings show a high prevalence of FSGS in comparison with most international renal pathological studies published in children. At the same time, there is a relatively low prevalence of IgA nephropathy as compared to some other Asian countries. Additional, more comprehensive, clinical and pathological studies on Saudi children with glomerular diseases are needed to confirm or negate these findings. Environmental and genetic factors have to be explored and studied for their role related to these differences. Also, there is an urgent need to establish a glomerulonephritis registry for children in Saudi Arabia. PMID- 18417808 TI - Percutaneous renal biopsy and its findings in children and adolescents in saudi arabia: a single center experience. AB - A retrospective study of 108 consecutive renal biopsies in children and adolescents below 18 years of age performed at the King Fahd Hospital, Jeddah, during an eight-year period ending 1996, was made. All the biopsies were performed by a single consultant nephrologist and under ultrasound guidance. Tru cut needles were employed in all cases and in the last two years, the bioptic gum was used. The age of the patients ranged between 3 months and 18 years with a mean of 10.6 years. There were 58 males and 50 females. All patients tolerated the biopsy well and there were no failures. The common side effects noted included gross hematuria in three (2.8%) and severe pain at the biopsy site in 11 (10.2%). None of the patients needed blood transfusions or prolonged hospitalization. Nephrotic syndrome was the commonest indication for performing the biopsy (83.3%) and among them, minimal change was the commonest lesion found (25%) followed by focal and segmental glomerulosclerosis (14.8%) and mesangial proliferative glomerulonephritis (15.7%). Our study further shows that renal biopsy is a safe procedure in children and the commonest indication is nephrotic syndrome. PMID- 18417809 TI - Chronic renal failure in children in asir region of saudi arabia. AB - Sixty two children aged 12 years or below, with chronic renal failure (CRF) were admitted to Asir Central Hospital, Saudi Arabia, in the period from June 1990 to June 1995. Of them, 39 (63%) were in CRF while 23 (37%) were in end-stage renal failure (ESRF) at the time of presentation giving a mean annual incidence for CRF of 15.6 and for ESRF of 9.2 per million children. The male to female ratio was 1.8:1. The mean age of the children was 5.5 years (age range 20 days to 12 years). Urinary system anomalies (including urinary tract anomalies) were the leading cause of CRF (64.5%) followed by gloraerulopathies (17.7%). In those who presented before the age of six years, SO.9% had CRF caused by urinary system anomalies (including inherited kidney anomalies such as polycystic disease) while such anomalies accounted for only 30% of CRF in children older than six years. Glomerulopathies were the leading cause (45%) of CRF in this latter group. We conclude that our mean annual incidence of CRF and ESRF is high and urinary system anomalies constitute the commonest cause among children in our region. Early diagnosis should therefore be encouraged to ensure early treatment of these anomalies, thus decreasing the incidence of CRF. PMID- 18417810 TI - CAPD in saudi arabian children: ten years experience from a single center. AB - In this paper we describe our experience with chronic ambulatory peritoneal dialysis (CAPD) in children. Between 1984 and 1993, a total of 51 patients were treated by CAPD at King Khalid University Hospital. Eight patients were excluded from the study because of incomplete data. There were 25 boys and 18 girls aged three months to 16 years; 12 patients (27.9%) were below two years, 15 patients (34.9%) were between 2-6 years, and 16 patients (37.2%) were above six years of age. Reflux nephropathy secondary to obstructive uropathy was the main cause of ESRD in the study patients (27.9%). The total period of CAPD was 579 patient months (48.25 years). The overall incidence of peritonitis was high, one episode per 4.7 patient months; one episode per 3.2 patient months in the first five years of the study and one episode per 6.2 patient-months in the 2nd five years of the study period. The cause of peritonitis was gram positive organisms in 38.7% and gram negative organisms in 24.9%. Seventeen (39.5%) of catheters were changed because of infection or mechanical obstruction. In 90.7% of patients, the mother was the person who was performing dialysis. At the end of the study, seven patients (16.2%) were still on CAPD, 10 patients (23.3%) shifted to heraodialysls, 16 patients (37.2%) were transplanted, six patients (14.0%) died and four patients (9.3%) recovered. We conclude that CAPD may be a suitable renal replacement therapy for children in a country with a wide geographical area and low population density as Saudi Arabia. PMID- 18417811 TI - Pediatric renal transplantation in saudi arabia. AB - This study was performed to evaluate the short and long-term outcome of pediatric renal transplantation in Saudi Arabia. Four large centers participated in this study and there were 124 patients with evaluable data. They included 79 males and 45 females with ages ranging from 2-18 years with a mean of 12.2 years. There were 69 transplants from living donors and 55 from cadaveric donors. The cause of the original renal disease was unknown in 61.4% of the cases. Re-transplantation rate was 8% and the cause of failure of the first graft was venous thrombosis in 60% of the cases most of whom were transplanted before 1990. Acute rejection occurred in 31% of the patients and did not correlate with age at time of transplantation, sex, nationality, source of donor, place of transplantation or follow-up, year of transplant, or delayed graft function; however, those who eventually lost their grafts had significantly more number of acute rejection episodes. There were 24 cases (19%) of infection related to graft and 43 cases (34%) of extra-renal infection. Medical complications other than rejection and infection occurred in 37% of the study group. The 15 year survival rates of patients and grafts from living donors were 96.7% and 92.2% respectively, while the 11 year survival rates of patients and grafts from cadaver donors were 93% and 58.4% respectively. We conclude that pediatric renal transplantation in Saudi Arabia has been performing well in terms of the quantity and quality and the results are comparable to the reputed programs in other countries. PMID- 18417812 TI - Spectrum of pediatric renal diseases in dubai. AB - A total of 712 patients with renal problems, aged 13 years or below (mean age 4.12 years) were seen in the Department of Health and Medical Services Hospitals in Dubai in the period from 1991 to 1996. The male to female ratio was 1:1.1. UAE citizens constituted 32% of the total, with a male to female ratio of 1:1.2. Nephrotic syndrome (26.3%) had the highest prevalence among the renal diseases seen, followed by urinary tract infection (19.1%), glomerulonephritis (GN) (9.7%), congenital renal anomalies (9.7%), and chronic renal failure (CRF) (7%). Congenital renal anomalies were the main cause of CRF in our patients followed by GN. Acute renal failure (ARF) occurred in 1.4% of the patients and was not an alarming problem; it had an uncomplicated course and good prognosis. Continuous ambulatory peritoneal dialysis was the mode of replacement therapy for patients with end-stage renal disease. Eight patients underwent renal transplantation; one cadaver donor, four living non-related donor (abroad) and three living related donor. PMID- 18417813 TI - Spectrum of pediatric renal diseases in jordan. PMID- 18417814 TI - Spectrum of pediatric renal diseases in Tunisia. PMID- 18417815 TI - Marching ahead with clinical trial registration. PMID- 18417816 TI - Statement on publishing clinical trials in Indian biomedical journals. PMID- 18417818 TI - An observational study of the proceedings of the All India Ophthalmological Conference, 2000 and subsequent publication in indexed journals. AB - AIMS: To determine the quality of reporting in the proceedings of the All India Ophthalmological Conference (AIOC) 2000, subsequent rate of publication in an indexed journal and differences between the proceedings and the journal version of these papers. DESIGN: Observational study. MATERIALS AND METHODS: All papers presented at the AIOC 2000 were retrieved from the proceedings and assessed for completeness of reporting. To determine the subsequent full publication, a Medline search was performed as of January 2007; consistency between the proceedings paper and the final publication was evaluated. STATISTICAL ANALYSIS: Chi square and Fisher's exact tests were used to compare publication rates based on geographical location, subspecialty and study design; Student's t -test was used to compare differences based on the number of authors and sample size. RESULTS: Two hundred papers were retrieved; many failed to include study dates, design or statistical methods employed. Thirty-three (16.5%) papers were subsequently published in indexed journals by January 2007. The published version differed from the proceedings paper in 27 (81.8%) instances, mostly relating to changes in author name, number or sequence. CONCLUSIONS: The overall quality of reporting of scientific papers in the proceedings of the AIOC 2000 was inadequate and many did not result in publication in an indexed journal. Differences between the published paper in journals and in proceedings were seen in several instances. Ophthalmologists should be cautious about using the information provided in conference proceedings in their ophthalmic practice. PMID- 18417817 TI - Diabetic retinopathy: an update. AB - Diabetes mellitus is a major cause of avoidable blindness in both the developing and the developed countries. Significant technological advances have taken place to improve the diagnostic accuracy of diabetic retinopathy. In the last three decades, the treatment strategies have been revised to include, besides laser photocoagulation, early surgical interventions and pharmacotherapies. PMID- 18417819 TI - Transconjunctival penetration of mitomycin C. AB - AIMS: The study was performed to estimate transconjunctival penetration of mitomycin C (MMC) to Tenon's tissue following application over the intact conjunctiva before routine trabeculectomy. SETTINGS AND DESIGN: Institution-based case series. MATERIALS AND METHODS: In 41 eyes of 41 patients, MMC (0.4 mg/ml for 3 min) was applied over the intact conjunctiva before beginning trabeculectomy. Tenon's capsule directly beneath the site of application was excised during trabeculectomy and was homogenized, centrifuged and MMC concentrations were analyzed using high-performance liquid chromatography (HPLC). STATISTICAL ANALYSIS USED: Statistical analysis was performed using STATA 8.0 version software (STATA Corporation, Houston, TX, USA). In this study, P -values less than 0.05 were considered as statistically significant. RESULTS: The average weight of the sample of Tenon's tissue excised was 5.51+/-4.42 mg (range: 0.9 17.1) and the average estimated MMC concentration found to be present in Tenon's tissue using HPLC was 18.67+/-32.36 x 10(-6) moles/kg of the tissue (range: 0.38 197.05 x 10(-6)). In 36 of the 41 patients (87.80%), the MMC concentration reached above 2 x 10(-6) moles/kg of the tissue concentration required to inhibit human conjunctival fibroblasts. CONCLUSIONS: Mitomycin C does permeate into the subconjunctival tissue after supraconjunctival application for 3 min. Application of MMC over the conjunctiva may be a useful alternative to subconjunctival or subscleral application during routine trabeculectomy and as an adjunct for failing blebs. PMID- 18417820 TI - Distinguishing infective versus noninfective keratitis. AB - For the purpose of this symposium, the term "keratitis" implies suppurative nonviral and viral keratitis. Corneal ulcers have been described in ancient literature. But even today, despite the availability of a wide range of newer antimicrobials and new diagnostic techniques, infective keratitis continues to pose a diagnostic and therapeutic challenge. This article focuses on the key diagnostic clinical features of the most common organisms causing infective keratitis - bacteria, fungi, viruses, nocardia and acanthamoeba - in India. While the clinical features in some cases are fairly straightforward, most cases challenge the clinician. We describe the salient clinical features which can help arrive at a diagnosis to begin appropriate treatment immediately, prior to the laboratory report. PMID- 18417821 TI - Investigative modalities in infectious keratitis. AB - Standard recommended guidelines for diagnosis of infectious keratitis do exist. Based on an extensive Medline literature search, the various investigative modalities available for aiding the diagnosis of microbial keratitis have been reviewed and described briefly. Preferred practice patterns have been outlined and the importance of routine pre-treatment cultures in the primary management of infectious keratitis has been highlighted. Corneal scraping, tear samples and corneal biopsy are few of the specimens needed to carry out the investigative procedures for diagnosis and for initiating therapy in cases of microbial keratitis. In bacterial, fungal and amoebic keratitis, microscopic examination of smears is essential for rapid diagnosis. Potassium hydroxide (KOH) wet mount, Gram's stain and Giemsa stain are widely used and are important for clinicians to start empirical therapy before microbial culture results are available. The usefulness of performing corneal cultures in all cases of suspected infectious keratitis has been well established. In cases of suspected viral keratitis, therapy can be initiated on clinical judgment alone. If a viral culture is needed, scrapings should directly be inoculated into the viral transport media. In vivo confocal microscopy is a useful adjunct to slit lamp bio-microscopy for supplementing diagnosis in most cases and establishing early diagnosis in many cases of non-responding fungal and amoebic keratitis. This is a non-invasive, high resolution technique which allows rapid detection of Acanthamoeba cysts and trophozoites and fungal hyphae in the cornea long before laboratory cultures give conclusive results. Other new modalities for detection of microbial keratitis include molecular diagnostic techniques like polymerase chain reaction, and genetic finger printing by pulsed field gel electrophoresis. PMID- 18417822 TI - Medical management approach to infectious keratitis. AB - This section provides guidelines on medical therapy of patients with infectious keratitis. In addition to initial empirical therapy, preferred medications, once the organisms responsible are isolated, are discussed. Atypical mycobacterial keratitis following lasik is described. General guidelines for supportive therapy and follow-up, of these patients are presented. Clinical response to treatment and indications for intervention are discussed. Possible causes and approach to cases refractory to medical therapy are discussed. PMID- 18417823 TI - First aid for complications of infectious keratitis. AB - Infectious keratitis is a fairly common entity in India. However while paying attention to the primary entity, the associated events may be overlooked. Enhanced pain usually suggests a worsening of the condition or development of associated problems like secondary glaucoma. However, contrary to logic, a sudden decrease in pain is also likely to suggest a worsening, e.g. perforation of the corneal ulcer. Various such problems with their management are outlined. PMID- 18417824 TI - Practical approach to medical management of glaucoma. AB - Primary open angle glaucoma (POAG) is usually a chronic, slowly progressive disease. At present, all resources are directed towards reduction of intraocular pressure (IOP), the only known causal and treatable risk factor for glaucoma, and medical management is frequently the first choice in most cases. With the introduction of innovative tools for early diagnosis and newer medications for treatment, decision-making in diagnosis and treatment of glaucoma has become more complex. The philosophy of glaucoma management is to preserve the visual function and quality of life (QOL) of the individual with minimum effects on QOL in terms of cost, side effects, treatment regime, follow-up schedules as well as socioeconomic burden. Our aim should be not to treat just the IOP, optic disc or visual field, but to treat the patient as a whole so as to provide maximum benefit with minimal side effects. In this article, we describe the scientific approach to medical management, mainly of POAG. PMID- 18417825 TI - Periocular necrotizing fasciitis associated with kerato-conjunctivitis and treated with medical management: a case report. AB - We report a 25-year-old systemically healthy male who presented with periocular necrotizing fasciitis (NF) in the left eyelid. This was associated with the presence of immunologically mediated marginal kerato-conjunctivitis, in the same eye. This potentially dangerous lid infection and the associated ocular surface infection resolved successfully, with medical management. We report this case to highlight the successful conservative management of periocular NF and the hitherto unreported anterior segment involvement. PMID- 18417826 TI - Calculating graft size and position in rotational corneal autografting: a simplified approach. AB - In eyes with eccentric corneal opacities partially involving the pupillary area, using a rotational corneal autograft, can help restore vision without the immunological complications associated with allografts. In this report, we describe a simple intraoperative method for determining trephine size and placement for rotational corneal autografting. This surgical approach helps in the planning and execution of rotational corneal autografting, to obtain good outcomes. PMID- 18417827 TI - Sudoriferous cyst of the orbit of adult origin after trauma. AB - A rare case of sudoriferous cyst of the orbit occurring in an adult, who had facial trauma, is reported. Several factors suggest its adult onset. The only other case reported in an adult is of presumed childhood origin. Very few congenital cases have been reported. A 65-year-old lady presented with recent onset of left-sided ptosis and a painless mass below the left supraorbital margin. The patient had traumatic ptosis after a road traffic accident 13 years ago. The ptosis was surgically repaired, which resulted in symmetrical palpebral apertures. Computed tomographic scan revealed a well-defined cystic mass in the anterior orbit. The mass was removed in toto by anterior orbitotomy. Histopathological examination revealed a single cyst lined by double-layered cuboidal epithelium in some areas and transitional epithelium at others. A periodic acid Schiff (PAS) positive, diastase-resistant glycocalyx lined the inner epithelium. Apical snouting suggested an apocrine nature. This confirmed a diagnosis of sudoriferous cyst. PMID- 18417828 TI - Recurrent neovascularization of the disc in sympathetic ophthalmia. AB - Sympathetic ophthalmia following parsplana vitrectomy is a known complication. We describe here a case of recurrent disc neovascularization in a patient of sympathetic ophthalmia. It promptly responded to steroids initially but later recurred with inflammation. PMID- 18417829 TI - Giant hanging melanoma of the eyelid skin. AB - Cutaneous melanoma of the eyelid is a rare entity. We present a 53-year-old male who had a nevus on the left upper eyelid skin since childhood, which transformed into a huge ulcerated hanging mass in the same region. Excision of the mass was done and histopathology confirmed the diagnosis of nodular malignant melanoma. A small preauricular lymph node showed metastatic melanoma on fine needle aspiration cytology. PMID- 18417830 TI - Descemet's membrane detachment caused by inadvertent vancomycin injection. AB - Descemet's membrane detachment is a condition with a wide range of etiologies. The most common cause is a localized detachment occurring after cataract surgery. We report a case of vancomycin injection-induced Descemet's membrane detachment as a complication following a routine cataract surgery and its management. PMID- 18417831 TI - Voriconazole for the treatment of refractory Aspergillus fumigatus keratitis. AB - We report a case of Aspergillus fumigatus keratitis in a 53-year-old, well controlled diabetic female who did not respond to standard antifungal treatment. She was started on topical natamycin eye drops, but the infiltrate continued to progress. Topical amphotericin B and systemic ketoconazole was added, however, there was no response and the infiltrate increased further. She was then switched to topical and systemic voriconazole. Steady resolution of the infiltrate was noted within 2 weeks of therapy. PMID- 18417832 TI - Acute orbital abscess complicating deep posterior subtenon triamcinolone injection. AB - A 54-year-old diabetic female presented with orbital abscess and corneal infiltrate 3 days after deep posterior subtenon triamcinolone acetonide injection in her right eye. This was administered immediately after focal laser photocoagulation for diabetic macular edema. The orbital abscess and corneal infiltrate responded to systemic and topical antibiotics. PMID- 18417833 TI - Transconjunctival orbital emphysema caused by compressed air injury: a case report. AB - Orbital emphysema following conjunctival tear in the absence of orbital wall fracture, caused by air under pressure is rare. Usually orbital emphysema is seen in facial trauma associated with damage to the adjacent paranasal sinuses or facial bones. To the best of our knowledge, there have been only eight reports of orbital emphysema following use of compressed air during industrial work. The air under pressure is pushed through the subconjunctival space into the subcutaneous and retrobulbar spaces. We present here a rare cause of orbital emphysema in a young man working with compressed air gun. Although the emphysema was severe, there were no orbital bone fracture and the visual recovery of the patient was complete without attendant complications. PMID- 18417834 TI - Isolated and silent spinal neurocysticercosis associated with pseudotumor cerebri. AB - Incidence of spinal neurocysticercosis (NCC) is rare. Isolated spinal NCC is still rarer. We present here a case report where a young lady presented with all the clinical features of pseudotumor cerebri (PTC), where medical treatment for PTC failed and the presence of cysticercous in spinal canal was detected only on the operation table, while doing a lumbo-peritoneal shunt (LP shunt) to save her vision. Diagnosis could be confirmed only after the histopathology report was received. She did not have any direct evidence of spinal involvement, thereby eluding correct diagnosis. In English literature, we could not find any report of isolated and silent spinal NCC associated with PTC. In addition, we could not find any report of recovery of cysticercous larva through the Touhey's needle injury, although this was an incidental finding. In endemic areas, isolated spinal NCC should be suspected in patients presenting with PTC. PMID- 18417835 TI - Right traumatic carotico-cavernous fistula with bilateral eye signs and post treatment right pseudo Argyll Robertson pupil. PMID- 18417836 TI - Sterilization of phacoemulsification handpieces. PMID- 18417837 TI - Debate on the various anti-vascular endothelial growth factor drugs. PMID- 18417838 TI - Primary 25-guage transconjunctival sutureless vitrectomy in pseudophakic retinal detachment. PMID- 18417839 TI - Intra-cameral injection of bevacizumab (Avastin) to treat anterior chamber neovascular membrane in a painful blind eye. PMID- 18417840 TI - Intravitreal bevacizumab (Avastin) for the treatment of proliferative sickle retinopathy. PMID- 18417841 TI - Immunohistochemistry in surgical pathology practice: a current perspective of a simple, powerful, yet complex, tool. AB - Immunohistochemistry (IHC) is a powerful tool in the surgical pathologists' armamentarium. The requests for IHC and the list of monoclonal antibodies have increased tremendously in the past decade. Issues concerning technical reproducibility, uniformity of interpretation, inter-laboratory comparability, and quality assurance are assuming greater importance due to the increased availability of IHC and its impact on diagnosis and therapy. An attempt has been made to give a current perspective of this simple and yet, in some aspects, a complex tool. PMID- 18417842 TI - Microwave histoprocessing versus conventional histoprocessing. AB - The aim of the study is to compare the histologic quality of the microwave histoprocessing with that of conventional method and to determine its positive impact on turnaround times and reduction of costs of tissue processing. One hundred and eighty-five paired tissue sections from different organs were taken. Each tissue sections were of size of 15 mm x 10 mm x 3 mm and divided into two; one set as experimental group and the other as control group. The tissues in the experimental group were further divided into six groups and processed by vacuum microwave method according to six protocols from I to VI. Other tissues in the control group were processed by the conventional method and compared. Overall, the quality of microscopic tissue from both the methods was identical. Microwave processing shortened the time of processing without compromising the overall quality of the histologic section and was cost-effective. PMID- 18417843 TI - A study of proliferative activity, angiogenesis and nuclear grading in renal cell carcinoma. AB - To evaluate the role of proliferative marker, proliferating cell nuclear antigen (PCNA) and microvessel density (MVD) as prognostic markers in renal cell carcinoma (RCC) and to see their relationship with the clinical stage and nuclear grades, we studied 30 cases of RCC for nuclear grading (Fuhrman's nuclear grade), MVD (using anti CD-34 antibody), and PCNA labeling index (using anti-PCNA antibody) over a period of 2.5 years. Staging was assessed by peroperative and radiologic findings. The area of highest MVD within the tumor was selected for microvessel count (MVC) per high-power field (0.1885 mm 2 area). PCNA labeling index was determined by counting percentage of positively stained tumor cell nuclei. PCNA labeling index above 60% was taken as high PCNA index and up to 60% was considered low. There was significant positive correlation between PCNA labeling index with both nuclear grade and clinical stage using Spearman's correlation coefficient. No association was noted between MVC with PCNA, nuclear grade, and clinical stages. Evaluation of proliferative status of RCC is a useful adjunct as a prognostic parameter as it is seen to correlate well with both clinical stage and nuclear grade. In our study, MVD was not seen to correlate with either of these. PMID- 18417844 TI - Gleason scoring of prostatic carcinoma: impact of a web-based tutorial on inter- and intra-observer variability. AB - A total of 40 cases of prostatic adenocarcinomas were scored independently by four pathologists using the Gleason scoring system. After attending a web-based tutorial, the scoring was repeated by all. Consensus scores were obtained by simultaneous viewing of each case in a multihead microscope by all four pathologists. The scores were then compared. The pretutorial kappa (kappa) values ranged from 0.36 to 0.64 with an average of 0.459. After the tutorial, the kappa values ranged from 0.44 to 0.678 with the average kappa value increasing to 0.538, thus indicating an improvement in the agreement. The intraobserver agreement ranged from 0.435 to 0.788. We conclude that web-based tutorials with emphasis on images developed by experts serve to achieve an uniformity in reporting. PMID- 18417845 TI - Clinicopathological study of non-neoplastic lesions of nasal cavity and paranasal sinuses. AB - An analysis of cases presenting as mass in nasal cavity (NC), paranasal sinuses (PNS), and nasopharynx (NP) was done over a period of 7 years in Jawaharlal Nehru Medical College, Aligarh. A provisional diagnosis was made after clinical assessment and radiological investigations, but final diagnosis was made after histopathological examination. The incidence of masses in NC, PNS, and NP was 34.3 cases per year, non-neoplastic lesions constituted 60% of these cases and their incidence was 20.7 cases per year. All the cases were carefully examined histopathologically and it was found that the region was affected by a variety of non-neoplastic lesions. Among 240 cases, 145 were non-neoplastic and 95 were neoplastic The lesions in the decreasing order of frequency were - nasal polyp, rhinoscleroma, tuberculosis, fungal infection, fibrous dysplasia, ossifying fibroma, cysts, nasal glioma, and cemento-ossifying fibroma. NP was involved by a range of neoplastic lesions; however, no non-neoplastic lesion was seen in this region. PMID- 18417846 TI - Primary cardiac sarcoma. AB - Primary cardiac sarcomas are uncommon entities. The diagnosis of sarcoma is not made preoperatively because of the rarity of the lesion and the nonspecific nature of the signs and symptoms. This is a report of a case of primary sarcoma arising from the left atrium, in a patient who presented with acute dyspnoea; the sarcoma resembled an atrial myxoma on echocardiography. PMID- 18417847 TI - Primary bilateral tubal adenocarcinoma associated with uterine leiomyomas. AB - Primary adenocarcinoma of the fallopian tube is the least common primary malignant tumor of the female genital tract. Bilaterality is also rare. Often the diagnosis is mistaken for ovarian tumor or tubo-ovarian mass. A case of bilateral primary tubal adenocarcinoma of serous type associated with uterine leiomyomas, without evidence of metastasis occurring in a postmenopausal woman is being reported. PMID- 18417848 TI - Sclerosing mucoepidermoid carcinoma with eosinophilia of the thyroid. AB - Sclerosing mucoepidermoid carcinoma with eosinophilia (SMECE) is a recently described carcinoma of the thyroid gland associated with Hashimoto's thyroiditis and considered to have a relatively indolent clinical course. We present a 65 year-old female, clinically euthyroid, who presented with a thyroid swelling since last 8 months along with right-sided cervical lymphadenopathy. Fine needle aspiration cytology suggested poorly differentiated carcinoma along with the involvement of the lymphnodes while histopathological examination showed features of SMECE with metastatic deposits in the right-sided cervical lymphnodes. PMID- 18417849 TI - Ovarian pregnancy. AB - Ovarian pregnancy is a rare form of extrauterine pregnancy contributing < 3% of ectopic pregnancies. We report an ovarian pregnancy in a 23-year female. PMID- 18417850 TI - Malignant hemangiopericytoma in the gluteal region: a rare entity. AB - We present here a case of malignant hemangiopericytoma in a 40-year-old female who presented with a history of slowly growing mass in left gluteal region for about 1.5 years. She was evaluated and a mass of 10 x 7.5 x 5.5 cm 3 was found on computed tomography. The tumor was resected and was sent for histopathological evaluation. Grossly the tumor was encapsulated and on cut section it was nodular. Microscopy showed spindle cells forming cords and sheets separated by thin delicate stroma consisting largely of thin-walled vessels revealed by reticulin stain. Pleomorphism and mitotic figures were seen. It was diagnosed as a case of malignant hemangiopericytoma of gluteal region. The patient underwent radiotherapy and did apparently well. She is on regular follow-up because long term follow-up is essential in all cases as recurrence can occur several years after treatment. PMID- 18417851 TI - Unusual presentation of dedifferentiated liposarcoma as paratesticular mass. AB - A 75-year-old male presented with complaint of painless left inguinoscrotal mass slowly increasing in size since 9 years. Clinical impression was malignancy arising in left testis or cord. Initial incisional biopsy was reported as fibromatosis. Later on wide excision of tumor was done and histopathology showed dedifferentiated liposarcoma with areas of fibromatosis. PMID- 18417852 TI - Sporotrichosis in Mysore: a case report to emphasize the role of histopathology. AB - Lymphocutaneous sporotrichosis is unusual in southern India. The diagnosis was made by histopathological examination which is purported to have poor sensitivity. The culture of the specimen confirmed the diagnosis. The pre-eminent role of a careful study of serial sections is emphasized. PMID- 18417853 TI - Melanocytoma of cerebellum. AB - Melanocytomas are rare melanocytic tumors of the central nervous system. We report a rare case of melanocytoma in the cerebellum of a 35-year-old male patient. Computed tomography scan showed a cystic lesion with an enhancing solid component arising from the cerebellum. The lesion was resected and a histological diagnosis of melanocytoma of the cerebellum was made. PMID- 18417854 TI - Brainstem abscess: the nonsurgical management. AB - The brainstem is an unusual location for a pyogenic abscess. Stereotactic aspiration or microsurgical drainage may be required in antibiotic refractory cases. Prolonged antibiotic therapy, along with symptomatic treatment may provide successful outcome. We report a case of cerebellar and brainstem abscess, managed successfully with prolonged antibiotic administration. PMID- 18417855 TI - Primary intraosseous meningioma. AB - Primary intraosseous meningioma of the skull is an uncommon lesion often confused preoperatively with a primary bone tumor of the skull. We report this rare tumor in a 19-year-old male who was radiographically diagnosed as osteoid osteoma. PMID- 18417856 TI - Angiomatous meningioma: a diagnostic dilemma. AB - Angiomatous meningioma accounts for 2.1% of all meningiomas. It has features of a typical benign meningioma with many small or large vascular channels which may predominate over its meningothelial elements. We present here a series of three cases of angiomatous meningioma, which posed diagnostic difficulty to clinicians, radiologists, and pathologists. All the three cases showed a tumor entirely composed of thin-walled vascular channels and cells with bland morphology in the background. The diagnosis was confirmed by immunohistochemistry. We present series of three cases to highlight the histomorphological features of this uncommon variant of meningioma that could help in distinguishing it from hemangioblastoma and hemangiopericytoma. PMID- 18417857 TI - Extraskeletal osteogenic sarcoma: a rare entity. AB - A 25-year-old female presented with a rapidly increasing painful swelling around right lower thigh and knee for the last 5 months with distant metastasis in the form of lung involvement. X-ray of the part showed only soft tissue swelling without any bony involvement. A diagnosis of extraskeletal osteosarcoma was made which was confirmed by immunohistochemistry. PMID- 18417858 TI - Osteochondroma-like parosteal osteosarcoma. AB - Parosteal osteosarcoma is a well-differentiated, predominantly fibro-osseous variant of osteosarcoma, accounting for 5% of all osteosarcomas. We report a case of parosteal osteosarcoma in the metaphyseal area of right femur, of 2 years' duration in a male aged 35 years. It was mistaken for osteochondroma in the initial biopsy, and the possibility of bizarre parosteal osteochondromatous proliferation (BPOP) was also considered. Subsequent excision of the tumor showed it to be a parosteal osteosarcoma. The patient had recurrence of the lesion after 1 year, and he attended a cancer institute. Follow-up showed metastases of the tumor on bone scan. PMID- 18417859 TI - Massive chondroid hamartoma of the lung clinically masquerading as bronchogenic carcinoma. AB - Chondroid hamartomas of the lung are uncommon lesions which are generally small sized and asymptomatic. Herein we describe a case of a large-sized pulmonary chondroid hamartoma which clinically mimicked bronchogenic carcinoma. A large hilar growth was detected in the left lung on radiological studies. Left upper lobectomy was done. The growth was well defined with a lobulated bluish cut surface. Histopathology disclosed lobules of mature cartilage rimmed by long slit like epithelial channels admixed with mature adipose tissue. PMID- 18417860 TI - Small-cell neuroendocrine tumor of larynx: a rare presentation. AB - Neuroendocrine tumors of the larynx are extremely rare and represent < 1% of all primary laryngeal tumors. We report a case of a 55-year-old man who presented to us with laryngeal tumor, histopathological evaluation of which revealed it to be a neuroendocrine tumor. Diagnosis is based mostly on light microscopy and may be supported by special histochemical stains in some instances. PMID- 18417861 TI - Primary tuberculosis of tongue. AB - Primary tuberculosis of tongue is very rare with unusual presentation creating a diagnostic dilemma. We report a case of primary tuberculosis of tongue in a 49 year-old female patient. Tuberculosis was not suspected clinically and there was no other focus elsewhere in the body. Fine needle aspiration cytology was attempted but was inconclusive. The diagnosis was made after histopathological examination. PMID- 18417862 TI - Inflammatory pseudotumor of lymph nodes presenting as pyrexia of unknown origin. AB - Inflammatory pseudotumor (IPT) is an uncommon benign disorder characterized by proliferation of spindle cells, inflammatory cells, and small vessels. The IPT of lymph nodes is a rare cause of lymphadenopathy that usually affects one or two nodal groups. We describe a 27-year-old male presenting with generalized lymphadenopathy, hepatosplenomegaly and fever for 1 year. Histologic examination of lymph node revealed few remnant lymphoid aggregates with marked sclerosis and numerous anastomosing blood vessels in lymph node parenchyma. Interspersed in between the fibrotic bands was a polymorphic infiltrate composed of lymphocytes, plasma cells, eosinophils, and immunoblasts. Also, many histiocytes, multinucleated giant cells some being Langhan's type and at places forming ill defined granulomas were observed. The presence of granulomas and Langhan's type of giant cells can lead to a misdiagnosis of tuberculosis as was also done in the present case. It is thus not only important to be aware of this rare benign pathologic entity while dealing with a case of lymphadenopathy, but also consider it in the differential diagnosis of granulomatous disease. PMID- 18417863 TI - Testicular retiform Sertoli cell tumor: a problem in histopathologic diagnosis. AB - A 9-year-old boy who presented with a left scrotal swelling was subsequently diagnosed as retiform sertoli cell tumor of testis which consisted entirely of retiform pattern. The case is reported in view of its rarity in this age group as well as its unique histologic appearance. PMID- 18417864 TI - Leiomyoma of scrotum. AB - Leiomyoma is a benign tumor of smooth muscles. Leiomyoma originating from the scrotum is a rare entity. We report here a case of 50-year-old male who presented with a 4-cm lump in the left side of scrotum. Clinically, it was provisionally diagnosed as sebaceous cyst and was excised. The histopathology showed findings consistent with leiomyoma. There was no cytological atypia or mitosis. The final diagnosis of solitary leiomyoma of scrotum was given. The patient was followed up regularly and is doing fine for last 4 months. PMID- 18417865 TI - Acid-fast bacilli positive isolated tubercular splenic abscess in a HIV-negative patient. AB - Isolated splenic tubercular abscess in a HIV-negative patient is an exceedingly rare clinical condition. It is even more rare to demonstrate acid-fast bacilli positivity on Ziehl-Neelson stains in such cases. We hereby present one such case. PMID- 18417866 TI - Endometrial stromal nodule with smooth muscle differentiation. AB - Uterine tumors composed of a prominent component of smooth muscle and endometrial stroma (so-called stromomyoma) are distinctly uncommon. This article describes the morphological features of one such tumor discovered as an incidental finding in a hysterectomy specimen of a 49-year-old lady with a clinical diagnosis of dysfunctional uterine bleeding. Morphological and immunohistochemical (IHC) evaluation were performed and a final diagnosis of endometrial stromal nodule with smooth muscle differentiation was rendered. PMID- 18417867 TI - Nodular regenerative hyperplasia of liver. AB - Nodular regenerative hyperplasia of the liver (NRHL) is a very rare cause of portal hypertension and liver failure. The condition is characterized by diffuse micronodular transformation of hepatic parenchyma without fibrous septa between the nodules. We present our experience with a 32-year-old woman who presented with recurrent episodes of upper gastrointestinal bleeding associated with massive splenomegaly who was subsequently found to have NRHL. This article considers the salient aspects of this rare condition, how it affects the patients and the options available in its management. A plea is made for the need for liver biopsy for all patients with portal hypertension especially those being considered for surgery. PMID- 18417868 TI - Coexistence of squamous cell carcinoma with dermoid cyst of ovary. AB - A 40-year-old female presented with pain and enlargement of abdomen since last 6 months. A mass was felt over right iliac fossa. Ultrasonography showed partly cystic and solid area in mass arising from ovary. A tooth-like structure was also seen. Dermoid cyst was the diagnosis offered. Laparotomy was performed. Ovarian mass with hair, a tooth and putty-like material was present. Solid white area with thickened wall was found. Histopathology confirmed the findings of dermoid cyst and the thickened area showed squamous cell carcinoma with areas showing keratinous material and giant cell reaction which was also seen at the periphery of tumor areas. Left ovary was normal and no deposits of tumor were seen. Postoperative period was uneventful. PMID- 18417869 TI - Sebaceous differentiation in odontogenic keratocyst. AB - Sebaceous glands are common in the oral mucosa, but are not normally present with in the jaws. A case of an odontogenic keratocyst with sebaceous glands in the epithelium in a 12-year-old child is presented here, with special emphasis on its histogenesis. PMID- 18417870 TI - Filarial granuloma in breast. AB - A case of filarial granuloma in breast in a 59-year-old female is reported. Adult worm was identified in a breast nodule. PMID- 18417871 TI - Malignant lymphoma presenting as nonhealing ulcer. PMID- 18417872 TI - Fine needle aspiration cytology of anterior mediastinal masses. AB - OBJECTIVE: The main aim of the study is to evaluate the computed tomography (CT) guided fine needle aspiration cytology (FNAC) of anterior mediastinal mass as a diagnostic procedure. MATERIALS AND METHODS: In all 135 cases, the material was obtained by CT-guided FNAC technique followed by staining with Papanicolaou and May-Grunwald-Giemsa stains. The histological material was obtained by needle biopsies, wedge biopsies and resection specimens. Immunohistochemical stains were used to confirm diagnosis in selected cases. RESULTS: Among 135 cases, cytohistology correlation was found in 92 cases. Correct typing was done in 53.33% cases. No correlation was found in 14.81% cases. Material was unsatisfactory in 18.51% cases. The diagnostic accuracy and positive predictive values were 85.71% and 78.26%, respectively. CONCLUSION: Although there are some limitations, most lesions of the anterior mediastinum can be diagnosed on FNAC. PMID- 18417873 TI - Cutaneous metastasis from follicular variant of papillary thyroid carcinoma: a case diagnosed on cytology. AB - A 40-year-old woman presented with a scalp swelling. A careful clinical examination revealed a left-sided deep-seated thyroid nodule. Fine needle aspiration from both sites disclosed a microfollicular architectural pattern on Giemsa stain and pale nuclei with nuclear grooves on Papanicolaou stain, indicating the possibility of follicular variant of papillary thyroid carcinoma with metastasis to the scalp. Histologic tissue evaluation confirmed the diagnosis. Thus, a diligent search for nuclear features should be performed to enable a diagnosis of follicular variant of papillary thyroid carcinoma on cytology. This helps in patient management obviating the need for a second surgical intervention. PMID- 18417874 TI - Microfilaria in thyroid gland nodule. AB - Presence of microfilaria in the thyroid gland is a rare finding. Filariasis is a common public health problem in the Indian sub-continent. Most of the cases of microfilaria in thyroid gland reported in the literature are associated with goiter and thyroid neoplasms. Here, we present a rare case that showed microfilaria on fine needle aspiration cytology of solitary thyroid nodule. PMID- 18417875 TI - Proliferative indices, cytogenetics, immunophenotye and other prognostic parameters in myelodysplastic syndromes. AB - Thirty-five adult myelodysplastic syndrome (MDS) patients were included in this study: 11 refractory anemia (RA), 4 RA with ring sideroblasts (RARS), 9 RA with excess of blasts (RAEB), 10 RAEB in transformation (RAEB-T) and 1 chronic myelomonocytic leukemia (CMML). The ranges of survival were 4-51 months, 40-59 months, 7-38 months, 5-24 months and 5 days, respectively. Three patients died and 3 showed disease progression during the course of the study. A composite analysis of proliferative indices, cytogenetics, immunophenotype and other conventional/novel prognostic parameters in the context of Indian MDS patients was performed. The proliferative indices (AgNOR and Ki 67 positivity), immunophenotypic markers, serum LDH and ferritin levels revealed wide variations and great overlap among different FAB subtypes. The scoring systems (Bournemouth, Dusseldorf and Goasguen) did not correlate with the prognosis and survival (p > 0.05). Clonal cytogenetic abnormalities were detected in 24/35 (68.57%) patients, +8, -5 and -7 being observed commonly. Cytogenetic abnormalities were more frequent in RAEB (88.8%), RAEB-T (80.0%) and RA (63.6%) subtypes of MDS. By Using Mufti's prognostic system and International prognostic scoring system (IPSS), a good positive correlation was found between low risk category and RARS with better survival as compared to other risk categories/FAB subtypes (p < 0.01). However, rest of the FAB subtypes were assigned into high, intermediate and low risk categories without any correlation with the survival and/or leukemic transformation. RARS subtype revealed itself as the better prognostic category according to the cytogenetic findings as well as Mufti's grading system. This was possible due to the longer follow up available for these patients (40-59 months). PMID- 18417876 TI - Serum transferrin receptor-ferritin index shows concomitant iron deficiency anemia and anemia of chronic disease is common in patients with rheumatoid arthritis in north India. AB - Anemia is a frequent cause of morbidity in patients with rheumatoid arthritis (RA). We studied the prevalence of anemia of chronic disorders (ACD) and ACD with coexistent iron deficiency anemia (IDA) in patients with RA using sTfR/log ferritin ratio (sTfR - F index). Complete blood counts, percent transferrin saturation, serum ferritin, sTfR, sTfR-F index measurements were carried out in 100 anemic RA patients. Twenty-five IDA subjects without any other illness and 25 age- and sex-matched normal controls were studied. Prevalence of anemia in RA patients was 50.5%. Patients with sTfR-F index value < 1.5 were classified as pure ACD and patients with sTfR-F index value> 1.5 were classified as ACD with coexistent IDA. Using these criteria, 20% patients were found to have pure ACD and 80% patients had coexistent ACD and IDA. In the normal control group, sTfR-F index was found to be 0.16-1.8. We found that sTfR-F index can clearly distinguish IDA control cases and normal subjects with no overlap in the range of sTfR-F index. PMID- 18417877 TI - Evaluation of iron status: zinc protoporphyrin vis-a-vis bone marrow iron stores. AB - Zinc protoporphyrin (ZPP) in the red cells is an indicator of iron status in the bone marrow (BM) and can be easily measured by Protofluor-Z Hematofluorometer from Helena Laboratories. It is well known that bone marrow iron is a gold standard for the diagnosis of iron deficiency anemia (IDA) even in the pre-latent phase. Hence, it was considered pertinent to evaluate the diagnostic utility of ZPP in comparison with bone marrow iron stores. 107 random BM were selected over a period of 2(1/2) years; in each case, RBC indices where recorded along with ZPP and Perls' Prussian blue reaction for BM iron stores. The specificity and sensitivity were found to be 77.8% and sensitivity 69.8%, respectively. However, the sensitivity increased up to 96.2% when Hb, RBC indices and ZPP were considered for the diagnosis of IDA. PMID- 18417878 TI - Comparative study of peripheral blood smear, quantitative buffy coat and modified centrifuged blood smear in malaria diagnosis. AB - The present study was aimed at modifying the centrifuged blood smear (modified centrifuged blood smear or MCBS), to make it a feasible and standardized procedure. The results obtained were compared with the current diagnostic methods - peripheral blood smear (PBS) and quantitative buffy coat (QBC). Blood samples collected from 100 suspected malaria patients were subjected to all three tests. It was found that PBS had 86.79% sensitivity and was absolutely specific. QBC was 96.22% sensitive and 93.61% specific. The majority of variations occurred in PBS negative cases; cases with parasite count Plasmodium falciparum. It was seen that by the addition of centrifugation to the conventional smear technique (MCBS) improved its sensitivity from 86.79% to near 100%. QBC and MCBS were found superior to PBS. Since MCBS combines principles of both QBC and PBS, it is as sensitive as QBC, as specific as PBS, and above all, easily performed and affordable. PMID- 18417879 TI - Splenic lymphoma with villous lymphocytes. AB - Splenic lymphoma with villous lymphocytes (SLVL) is a rare disorder that comprises less than 1% of lymphoid neoplasms. It is the leukemic counterpart of splenic marginal zone lymphoma (SMZL) and is characterized by splenomegaly, often with no lymphadenopathy, moderate lymphocytosis and villous lymphocytes on peripheral blood smear. Here, we report a case of SLVL in a 56-year-old male with very high leukocyte counts, massive splenomegaly and relatively few leukemic cells with subtle villous projections on the surface. This disorder is often confused with other chronic lymphoproliferative disorders, especially chronic lymphocytic leukemia (CLL) and hairy cell leukemia and should be differentiated from them. We are reporting this case to highlight the diagnostic pitfalls associated with this disorder. PMID- 18417880 TI - Granular acute lymphoblastic leukemia in an adult patient. AB - Granular acute lymphoblastic leukemia (G-ALL) may mimic the diagnosis of acute myeloid leukemia due to the presence of cytoplasmic granules found in the lymphoblasts. This rare variant is commonly seen in children but may occur in adults also. We report a case of G-ALL in an adult female patient. PMID- 18417881 TI - Simultaneous appearance of dual malignancies of hematopoietic system-multiple myeloma and acute myeloid leukemia. AB - We herein report a case of denovo and simultaneous appearance of multiple myeloma and acute myeloid leukemia in a 58-year-old female patient, without prior exposure to chemotherapy or radiotherapy. This case is reported because of its extreme rarity. PMID- 18417882 TI - Morphological spectrum of AIDS-related plasmablastic lymphomas. AB - We have had a recent spurt in cases of AIDS-related lymphoma (ARL) at our centre. Most of these cases are aggressive mature B cell lymphomas, mainly plasmablastic lymphoma (PBL) and diffuse large B-cell lymphoma (DLBCL). Most of the PBL are extranodal in location and are mucosa-based. We reviewed the morphological features of 34 cases of PBL. Diagnosis was based on morphology, immunohistochemistry, proliferation index, HIV positive status and its preference to extranodal sites (mostly mucosa based). We classified PBL into three morphological subtypes (immunoblastic - 25, Burkitt's - 7, plasmacytic - 2). Tumor cells expressed as leucocyte common antigen (LCA) in 60%, CD138 in 100%, EMA in 45% and light chain restriction in 86% cases. CD20 was negative in all cases. Pathologists need to be aware of PBL and its various morphological subtypes as the identification of this entity from its close differentials carries major therapeutic implications. PMID- 18417883 TI - Hemophagocytic syndrome. PMID- 18417884 TI - Enumeration of CD4 and CD8 T lymphocytes in healthy HIV seronegative adults of northwest India: a preliminary study. AB - CD4 T lymphocyte count is used to measure the progression of HIV infection and to monitor the response to antiretroviral therapy. Information on reference CD4 and CD8 T cell counts in healthy individuals is lacking in northwest India. Samples from 65 HIV-seronegative healthy volunteers (males, 37; females, 28) aged 18 through 59 years were analyzed using FACS (Fluorescent Antibody Cell Sorter) Count TM System. The values of mean and standard deviation of each lymphocyte subpopulation were estimated. The mean +/- SD of absolute numbers of CD4 and CD8 lymphocytes/microl was 743.4 +/- 307.8 and 541.7 +/- 176.4 in males and 790.7 +/- 280.4 and 497.03 +/- 203.6 in females respectively. The range of CD4 counts was 379 to 1800 in males and 321 to 1265 in females. The mean CD4:CD8 ratio was 1.43 +/- 0.56 in males and 1.78 +/- 0.76 in females. The results of this study show a wide variability in CD4 counts in the Indian population. A large multicentric study would define normal ranges of CD4, CD8, and CD4:CD8 ratios among the Indian population. PMID- 18417885 TI - The prevalence of extended-spectrum beta-lactamase in environmental isolates of Enterobacter. AB - The incidence of extended-spectrum beta-lactamase (ESBL)-producing strains and multidrug-resistant strains of Enterobacter spp. isolated from the 1312 km long river Narmada was investigated. Out of the 57 isolates of Enterobacter, 73.68% were found to be ESBL producers including the isolates of E. taylorae and isolates of E. agglomerans, which have been characterized for the first time. All the isolates were found susceptible to the antibiotic imipenem. AmpC gene was found in all the Enterobacter strains tested. AmpC beta-lactamase-producing bacterial pathogens may cause major therapeutic failure if not detected and reported in time. It was seen that these enzymes are mainly chromosomally mediated along with several non-AmpC beta-lactamase. PMID- 18417886 TI - Evaluation of an enzyme-linked immunoassay for the detection of Cryptosporidium antigen in fecal specimens of HIV/AIDS patients. AB - Cryptosporidium parvum, a protozoan parasite, causes severe diarrhea in immunodeficient hosts like HIV/AIDS patients, leading to significant morbidity and mortality. Diagnosis of the Cryptosporidium oocyst in the stool of these patients by conventional microscopy is labor intensive and time consuming. Therefore, we planned to evaluate the usefulness of a stool ELISA test in detecting Cryptosporidial antigen. About 89 stool specimens obtained from HIV seropositive patients with diarrhea were subjected to an ELISA test and modified acid-fast staining (gold standard), on both direct and formol ether-concentrated specimens. The prevalence of Cryptosporidial diarrhea was found to be 12.4% (11/89). Other enteric pathogens detected were Isospora belli (3), Giardial cyst (3), Entamoeba coli cyst (2), and Entamoeba histolytica cyst (1). Dual infection with Cryptosporidium and Isospora belli was seen in two patients. Concentration technique improved identification by microscopy. The sensitivity and specificity for stool ELISA were found to be 90.9% and 98.7% respectively. The results of stool ELISA indicate that this simple, rapid, reliable, and standardized immunoassay test is sensitive and specific for routine diagnosis and may be useful for large-scale epidemiological studies of Cryptosporidiosis. PMID- 18417887 TI - Prevalence of extended-spectrum beta-lactamases among Escherichia coli and Klebsiella pneumoniae isolates in a tertiary care hospital. AB - Extended-spectrum beta-lactamases (ESBLs) continue to be a major problem in clinical setups the world over, conferring resistance to the expanded-spectrum cephalosporins. Knowledge about their prevalence is essential to guide towards appropriate antibiotic treatment. The aim of the present study is to determine the prevalence of ESBL producers among Escherichia coli and Klebsiella pneumoniae isolates at a tertiary care institution. A total of 357 clinical isolates comprising E. coli (n = 181) and K. pneumoniae (n = 176) were recovered from various clinical samples over a period of six months from April to September 2006. Antibiogram profile of these isolates was determined to commonly used antibiotics, along with screening for ESBL production by the screening test as recommended by the Clinical Laboratory Standards Institute (CLSI). Isolates which showed positive results with screening test were shortlisted for confirmatory tests of ESBL production. Two tests were performed: phenotypic confirmatory test with combination disk and the minimum inhibitory concentration (MIC) reduction test. Out of 357 isolates of E. coli and K. pneumoniae screened for ESBL production, 120 were found to be potential ESBL producers. Of these, 80 isolates were confirmed to be ESBL producers. Thus the prevalence of ESBL-producing isolates of E. coli and K. pneumoniae was found to be 22% (80 out of 357). This was significantly lower than the data available from other hospitals. PMID- 18417888 TI - Morbidity pattern of hydatid disease (cystic echinococcosis) and lack of its knowledge in patients attending Mamata General Hospital, Khammam, Andhra Pradesh. AB - There is hearsay that prevalence of hydatid disease in Khammam and Nalgonda districts of Andhra Pradesh is high. We report here a preliminary study conducted to determine the magnitude of the problem of hydatid disease and the morbidity associated with it in patients attending MGH, KMM, A.P. (rural hospital). Eleven cases were identified during the period from November 2005 to May 2006 (seven months). Pain in abdomen, mass per abdomen, loss of appetite, pregnancy complicated by cystic echinococcosis (CE), and jaundice were the main clinical symptoms and signs. Ultrasonography, detection and removal of the cysts on the operation table, microscopic examination of the aspirated hydatid fluid were confirmatory. Ziehl-Neelsen stain of the aspirated fluid revealed acid-fast scolices. Interrogation of the patients and their family members (50) revealed that there was a total lack of knowledge of dog-tapeworm-caused infection in humans. They knew 'rabies' as the only disease man gets from dogs, and tapeworms are from pork and beef. PMID- 18417889 TI - Ocular thelaziasis in Assam: a report of two cases. AB - Small, white, thread-like, motile worms were recovered from the conjunctival sac of a 13-year-old girl and a 50-year-old woman from Dibrugarh district, Assam, India. They were identified as thelazia species. These two cases have been reported due to their rarity in India and elsewhere. PMID- 18417890 TI - Hymenolepis diminuta in a child from rural area. AB - We report a rare case of Hymenolepis diminuta infection in a 12-year-old girl from a rural area of Devghar. PMID- 18417891 TI - Nocardia brain abscess in a diabetic patient. AB - Brain abscess due to disseminated nocardia infection is an acute medical emergency among immunocompromised patients. We report a case of rapidly progressive nocardia brain abscess in an apparently healthy diabetic individual. The close similarity of the radiological features with those of malignancy and tuberculosis may delay the diagnosis of central nervous system (CNS) nocardiosis. A high index of suspicion and early intervention like stereotactic brain biopsy remain the cornerstone to increase the chance of positive clinical outcome. PMID- 18417892 TI - Incidence and changing pattern of mycetoma in western Rajasthan. AB - Histopathologic analysis of 73 cases of mycetoma occurring in western Rajasthan was done from January 2001 to December 2005. Maduromycotic mycetoma remains commonest in this region as compared to actinomycotic mycetoma, which is more common in the southern part of Rajasthan. The incidence of actinomycotic mycetoma has increased during the last five years in this part of Rajasthan due to changes in climatic conditions, like heavy rainfall, increased irrigation by Rajasthan Canal, urbanization of villages, and modification in agriculture. The ratio of prevalence of maduromycotic mycetoma to the prevalence of actinomycotic mycetoma has decreased from 4:1 to 1.91:1 during the last five years in western Rajasthan. PMID- 18417893 TI - Criteria for deciding cost-effectiveness for expensive new anti-cancer agents. PMID- 18417894 TI - The effect of three mouthwashes on radiation-induced oral mucositis in patients with head and neck malignancies: a randomized control trial. AB - AIMS: The present study was done to assess the effect of three alcohol-free mouthwashes on radiation-induced oral mucositis in patients with head and neck malignancies. MATERIALS AND METHODS: Eighty patients with head and neck malignancies, scheduled to undergo curative radiotherapy, were randomly assigned to receive one of the three alcohol-free test mouthwashes (0.12% chlorhexidine, 1% povidone-iodine, or salt/soda) or a control. The patients were instructed to rinse with 10 ml of the mouthwash, twice a day, for a period of 6 weeks. Mucositis was assessed at baseline and at weekly intervals during radiation therapy, using the World Health Organization criteria for grading of mucositis. The baseline demography of the four groups was matched for age, sex, stage of cancer, and whether the patient had cancer of oral or extraoral regions. A post hoc test for repeated measures was used to find the difference of mean mucositis scores between the groups at various week intervals. RESULTS: Among the 76 patients who completed the study, patients in the povidone-iodine group had significantly lower mucositis scores when compared to the control group from the first week of radiotherapy. Their scores were also significantly lower when compared to the salt/soda and chlorhexidine groups from the fourth and fifth week, respectively, after radiotherapy. CONCLUSIONS: This study demonstrates that use of alcohol-free povidone-iodine mouthwash can reduce the severity and delay the onset of oral mucositis due to antineoplastic radiotherapy. PMID- 18417895 TI - Implications of contrast-enhanced CT-based and MRI-based target volume delineations in radiotherapy treatment planning for brain tumors. AB - Delineation of various target volumes using contrast-enhanced magnetic resonance imaging (MRI) and/or computed tomography (CT) constitutes the primary step for radiation therapy planning (RTP) in brain tumors. This study presents a quantification and comparative evaluation of the various clinical target volumes (CTV) and gross target volumes (GTV) as outlined by contrast-enhanced CT and MRI, along with its implications for postoperative radiotherapy of brain tumors. Twenty-one patients of gliomas were considered for this prospective study. Peritumoral edema as CTV and residual tumor as GTV were delineated separately in postoperative contrast-enhanced CT and MRI. These volumes were estimated separately and their congruence studied for contrast-enhanced CT and MRI. Compared to MRI, CT underestimated the volumes, with significant differences seen in the mean CTV (mean +/- SD: -62.92 +/- 93.99 cc; P = 0.006) and GTV (mean +/- SD: -21.08 +/- 36.04 cc; P = 0.014). These differences were found to be significant for high-grade gliomas (CTV: P = 0.045; GTV: P = 0.044), while they were statistically insignificant for low-grade gliomas (CTV: P = 0.080; GTV: P = 0.117). The mean differences in the volumes for CTV and GTV were estimated to be 106.7% and -62.6%, respectively, taking the CT volumes as the baseline. Thus, even though, electron density information from CT is essential for RTP, target delineation during postoperative radiotherapy of brain tumors, especially for high-grade tumors, should be based on MRI so as to avoid inadvertent geographical misses, especially in the regions of peritumoral edema. PMID- 18417896 TI - Radiofrequency ablation of hepatic metastasis: results of treatment in forty patients. AB - AIM: To evaluate the local control of hepatic metastasis with radiofrequency ablation treatment. MATERIALS AND METHODS: We did a retrospective analysis in 40 patients treated with radiofrequency ablation for hepatic metastasis. The tumors ablated included up to two metastatic liver lesions, with primaries in breast, gastrointestinal tract, cervix, etc. Radiofrequency ablation was performed under general anesthesia in all cases, using ultrasound guidance. Radionics Cool-Tip RF System was used to deliver the treatment. RESULTS: The median age of patients treated was 49 years. There were 13 female and 27 male patients. The median tumor size ablated was 1.5 cm (0.75-4.0 cm). A total of 52 radiofrequency ablation cycles were delivered. Successful ablation was achieved in all patients with hepatic metastasis less than 3 cm in size. Pain was the most common complication seen (75%). One patients developed skin burns. At 2-year follow-up 7.5% of patients had locally recurrent disease. CONCLUSIONS: Radiofrequency ablation is a minimally invasive treatment modality. It can be useful in a select group of patients with solitary liver metastasis of less than 3 cm size. PMID- 18417897 TI - Execution of mantle field with multileaf collimator: a simple approach. AB - BACKGROUND: Until very recently mantle field radiotherapy remained the gold standard for the treatment of favorable early-stage Hodgkin's lymphoma. The classic mantle includes all the major lymph nodes above the diaphragm and extends from the inferior portion of the mandible to the level of the insertion of the diaphragm. AIMS: To describe a simple technique that has been devised to treat the mantle field with the help of multileaf collimator and using computed tomography (CT)-based treatment planning. MATERIALS AND METHODS: CT scan was performed with the patient in the supine position and the datasets were transferred to the Eclipse treatment planning system. Elekta Precise linear accelerator equipped with 40 pairs of multileaf collimator (MLC) was used for the execution of the mantle field. The MLC's shapes were designed to take the shape of the conventional customized blocks used for treatment of mantle field. The anterior mantle field was divided into three separate MLC segments with the collimator kept at 0 degrees. The first MLC segment was shaped to cover the neck, clavicular regions, and mediastinum. The second and the third MLC segments covered the right and left axilla, respectively. The posterior fields were opposed to the anterior subfields in a similar fashion. The dose was prescribed at the midplane, using reference points. RESULTS AND CONCLUSION: The technique described in this study is very simple, easy to implement, and avoids unnecessary delay in the execution of the mantle field. The mantle field can be easily shaped with the multileaf collimators, without any collimator rotation. PMID- 18417898 TI - Prognostic and diagnostic value of serum pseudocholinesterase, serum aspartate transaminase, and serum alinine transaminase in malignancies treated by radiotherapy. AB - BACKGROUND: There is substantial evidence that environmental factors cause or accelerate the onset of malignancy. Environmental factors, due to the presence of many pollutants and carcinogenic agents, alter cellular growth, which leads to biochemical changes in the blood. MATERIALS AND METHODS: In the present study, we estimated serum pseudocholinesterase (PCHE), serum aspartate transaminase (AST), and serum alanine transaminase (ALT) in 92 patients with head and neck cancer and 71 patients with cancer of the uterine cervix; all of them were patients attending our department for radiation therapy. We also estimated PCHE, AST, and ALT levels in 30 healthy normal individuals. The estimations in cancer patients were done before the start of radiotherapy, midway through radiotherapy (30 Gy dose), at the end of radiotherapy, and during subsequent monthly follow-up visits over a period of at least 6 months. RESULTS AND DISCUSSION: We have observed that the PCHE levels were lower (31-49% of normal value) in all patients with malignancies, except in those with stage II head and neck cancers. We also found that the levels start increasing as radiotherapy progresses. The patients with no detectable/visible disease activity at 6 months follow-up showed PCHE values in the normal range. Similarly, the AST and ALT values were much higher (138-229% of normal value) in all the malignant cases as compared to the normal healthy individuals. The values decrease and approach normal levels as radiotherapy progresses and, in 92% of head and neck cancer cases [stages IIA, IIB, and IIIA] with no disease activity, the PCHE, AST, and ALT were normal or near normal; the corresponding figure in cancer cervix cases was 89%. From the present study we conclude that PCHE, AST, and ALT can be used as good prognostic biochemical tumor markers in the management of malignancies of the head and neck and uterine cervix. PMID- 18417899 TI - An overview on applications of optical spectroscopy in cervical cancers. AB - Despite advances in the treatment modalities, cervical cancers are one of the leading causes of cancer death among women. Pap smear and colposcopy are the existing screening methods and histopathology is the gold standard for diagnosis. However, these methods have been shown to be prone to reporting errors, which could be due to their subjective interpretation. Radiotherapy is the mainstay of treatment for the locally advanced stages of cervical cancers. The typical treatment regimen spans over 4 months, from the first fraction of radiation to clinical assessment of tumor response to radiotherapy. It is often noticed that due to intrinsic properties of tumors, patients with the same clinical stage and histological type respond differently to radiotherapy. Hence, there exists a need for the development of new methods for early diagnosis as well as for early prediction of tumor radioresponse. Optical spectroscopic methods have been shown to be potential alternatives for use in cancer diagnosis. In this review, we provide a brief background on the anatomy and histology of the uterine cervix and the etiology of cervical cancers; we briefly discuss the optical spectroscopic approach to cervical cancer diagnosis. A very brief discussion on radiation therapy and radiation resistance is also provided. We also share our experiences with the Raman spectroscopic methodologies in cervical cancer diagnosis as well as in the prediction of tumor radioresponse. PMID- 18417900 TI - Radiotherapy for management of skin cancers in fibrodysplasia ossificans progressiva: a case report and review of the literature. AB - Fibrodysplasia ossificans progressiva (FOP) is a rare condition of ectopic calcification leading to increasing disability throughout life, with most patients being wheelchair bound by the age of 30. Ectopic calcification can be triggered by trauma and it is therefore important to minimize biopsies and operative procedures in affected individuals. We report a 46-year-old FOP patient who was successfully treated with radiotherapy for a basal cell carcinoma. There are no previous reports in the literature on the management of skin malignancies in these patients and very limited literature on outcome following external beam radiotherapy. PMID- 18417901 TI - Sarcomatoid squamous cell carcinoma of uterine cervix: pathology, imaging, and treatment. AB - Sarcomatoid squamous cell carcinoma of the cervix is a rare tumor. Only 16 cases have so far been reported in literature. We report here one such tumor occurring in a 54-year-old postmenopausal woman. Our case report describes the clinical, pathological, and PET scan characteristics of this tumor. The patient was treated with concurrent chemoradiotherapy and is disease free at 6-months follow-up. PMID- 18417902 TI - Chest wall metastasis from hepatocellular carcinoma in the absence of a primary: An unusual presentation. AB - Metastatic hepatocellular carcinoma (HCC) has an aggressive course with a very poor outcome. The common hematogenous metastatic sites are the lungs, bones, and adrenal glands. The chest wall is an extremely rare site of metastasis from HCC. We report a rare presentation in a gentleman, where the chest wall metastasis kept progressing in spite of treatment, without any evidence of a detectable primary. PMID- 18417904 TI - Accelerated partial breast irradiation: an advanced form of hypofractionation. AB - Altered fractionation schedules are being increasingly investigated in the treatment of breast cancer. Two such schedules that are frequently compared are hypofractionated external beam radiation therapy (HERT) and accelerated partial breast irradiation (APBI). Though these two modalities are considered separately, APBI is an actually an advanced form of hypofractionation, where acceleration of the treatment is possible due to the smaller volume being irradiated. HERT as well as APBI are investigational at present and are being tested in randomized trials. This article looks at the advantages of APBI as a hypofractionation schedule. PMID- 18417903 TI - Endobronchial metastasis of follicular thyroid carcinoma presenting as hemoptysis: a case report. AB - Endobronchial metastasis secondary to follicular thyroid carcinoma is extremely rare. Here, we report a case of follicular thyroid cancer in 58-year-old male who presented with hemoptysis. Computed tomography of the chest revealed multiple lung metastases. Flexible fiberoptic bronchoscopy revealed a fragile polypoid mass 5 cm distal to the vocal cords; biopsy taken from this mass revealed follicular thyroid carcinoma. PMID- 18417905 TI - Coexistence of carcinoma breast and Paget's disease of bone. AB - We report a case of a patient with carcinoma breast who was incidentally diagnosed to be also suffering from Paget's disease of bone on a routine radionuclide bone scan. CT-guided biopsy and histopathology later confirmed the diagnosis. PMID- 18417906 TI - Dysplastic hematopoiesis and underlying dysthyroidism. PMID- 18417907 TI - Malignancy in renal transplant recipients. AB - Immunosuppressed organ allograft recipients have a 3-4 fold increased risk of developing cancer, but the chance of developing certain malignancies is increased several hundredfold. With the exception of skin cancers, most of the common neoplasms seen in the general population are not increased in incidence in organ allograft recipients. Instead, there is a higher frequency of relatively rare tumors including lymphomas, Kaposi's sarcoma, other sarcomas, vulvar and perineal carcinomas, renal and hepatobiliary carcinomas. Tumors appear after a relatively short time post-transplantation. The earliest is Kaposi's sarcoma, which appears after an average of 22 months post-transplantation, and the latest are vulvar and perineal carcinomas, which present after an average of 113 months post transplantation. Unusual features of lymphomas are: (a) high incidence of non Hodgkin's lymphomas; (b) high frequency of Epstein-Barr virus-related lesions; (c) frequent involvement of extra-nodal sites; (d) marked predilection for the brain; and (e) frequent allograft involvement. Skin cancers also present unusual features: (a) remarkably high frequency of Kaposi's sarcoma; (b) reversal of the ratio of basal to squamous cell carcinomas seen in the general population; (c) young age of the patients; and (d) high incidence of multiple tumors, which occur in 43% of patients. Vulvar and perineal cancers occur at a much younger age than in the general population. Probably, multiple factors play a role in the etiology of the cancers. Immunodeficiency per se and infection with oncogenic viruses may be major influences. Other factors possibly playing a role include direct damage to DNA by various immunosuppressive agents; possibly synergistic effects of these treatments with carcinogens; and genetic factors influencing susceptibility or resistance to development of malignancy. PMID- 18417908 TI - Tuberculosis in hemodialysis patients. PMID- 18417909 TI - The Effect of Short Term Use of Indomethacin, a Non-Steroidal Anti--inflammatory Drug, on Peritoneal Dialysis Patients. AB - In this study, the effect of indomethacin a nonsteroidal anti-inflammatory drug (NSAID) was studied to evaluate the effect of its short-term use on peritoneal clearance of urea and creatinine as well as protein excretion in nine chronic renal failure patients on intermittent peritoneal dialysis therapy. Four of them were males and five were females with mean age of 54.4 years. Clearance values and protein excretion in the dialysate effluents were measured before and after administering the indomethacin. There was no significant effect on the urea and creatinine clearance after the use of this drug. However, the mean +/- SE protein concentration in the dialysate effluents decreased significantly after the use of indomethacin from 43.1 + 4.5 to 37.0 + 3.5 mg/dl (p< 0.01). This reduction of protein excretion in the dialysate after the use of indomethacin might enhance the nutritional status of patients on peritoneal dialysis. PMID- 18417910 TI - Is peritonitis a limiting factor in the widespread use of CAPD in the developing countries? AB - Although continuous ambulatory peritoneal dialysis (CAPD) is now an established form of treatment for patients with end-stage renal failure (ESRF), it is not being used on a large scale in developing countries (DC). One of the limiting factors in this regard is peritonitis which is the most common complication of CAPD. Data on 70 patients on CAPD in our center were analyzed to study the impact of peritonitis on long-term outcome of this treatment. The mean age of the study patients was 37.2 years and the mean follow-up was 32.8 months per patient. A total of 41 patients (58.6%) received CAPD as first choice, 27 (38.6%) were started on CAPD after being on hemodialysis earlier and two (2.8%) were started on CAPD after a failed renal transplantation. Thirty three patients (47.1%) used safe-leur lock system, 23 (32.9%) used the spike system and 14 (20%) used safe leur system initially and subsequently were changed to spike system. The overall incidence of peritonitis encountered was one episode per 32.3 patients months and was higher among patients who started CAPD as second choice. The incidence of peritonitis was not influenced by sex, age, profession or presence of diabetes mellitus or cardiovascular disease. This study shows that peritonitis rate in CAPD patients in our hospital is low and compares favorably with results from established centers in the developed countries. Thus, worries about recerrent peritonitis should not hinder CAPD programs in DC. PMID- 18417911 TI - Tuberculosis in patients on maintenance hemodialysis: a single center experience. AB - This retrospective study was performed to review the prevalence, clinical presentation and outcome of tuberculosis (TB) in patients on maintenance hemodialysis (HD) as compared to that of the general population in Jordan. The records of 927 patients who were maintained on maintenance HD at King Hussein Medical Center between 1986 and 1993 were reviewed for the development of TB. There were 550 males and 377 females in the study group. Eleven of these patients (1.2%) were diagnosed to have TB during this period. This figure is higher than the prevalence rate of 0.015% reported among the general population in some regions of Jordan. Of the 11 patients with TB, there were four males and seven females. Their ages varied from 28 to 70 years and the duration on maintenance HD before the onset of TB ranged between 1 and 24 months. Extra-pulmonary manifestations predominated (81.8%) and TB lymphadenitis was the commonest lesion observed. Five patients died during the study period but none due to a direct consequence of TB or its treatment. We conclude that the prevalence of TB is high among patients on HD In Jordan when compared with the general population. PMID- 18417912 TI - Vesico ureteric reflux and renal scarring in children with urinary tract infection. AB - To evaluate the presence of vesico ureteric reflux (VUR) and renal scarring in children with documented symptomatic urinary tract infection (UTI), and the importance of age at time of presentation, we studied 29 patients with UTI. Nineteen patients were females and 10 were males. Three patients were below one year of age, 13 were between one and five years, and 13 were above five years. None of the patients had systemic hypertension or renal insufficiency. All patients had renal ultrasound and renal scintigraphy with dimercapto succinic acid (DMSA). Micturating cystourethrogram was done in all children below age of five, and in patients above age of five if renal scintigraphy showed evidence of scarring. Of the 29 UTI patients 10 (34%) had VUR. Of the patients with VUR, seven were below age of five while three were above five. Renal scarring was found in 3/7 patients with VUR below age of five years, but was in all patients with VUR above age of five years. All patients with grade 3 and 4 reflux nephropathy had renal scarring, while it was found in half of the patients with grade 1 and 2 reflux. We conclude that in UTI patients VUR is not uncommon finding, which needs further investigations and follow-up. PMID- 18417913 TI - Aluminum monitoring for chronic renal failure patients in kuwait. AB - The importance of monitoring serum aluminum (Al) level in patients undergoing long-term hemodialysis (HD) is well known. This study on 94 HD patients from Mubarak Hospital, Kuwait revealed that serum Al level was significantly higher in HD patients on aluminum hydroxide therapy (n = 57) than those not on this treatment (n = 37) (p = 100 ug/L. Analysis of water and dialysis fluid showed Al levels just above the permissible limit, on certain occasions, suggesting the need for regular monitoring of these substances for Al contamination. Patients on HD for longer than five years and those above the age of 50 years had higher serum Al levels. The most important single factor causing elevated serum Al levels in the study patients was aluminum hydroxide therapy, administered orally as a phosphate binder. PMID- 18417914 TI - Pefloxacin in the treatment of childhood nephrotic syndrome: a case report. AB - The idiopathic nephrotic syndrome (INS) of childhood is characterized chiefly by a remitting and relapsing course and its striking susceptibility to corticosteroid therapy. We report a case of relapsing nephrotic syndrome associated with urinary tract infection (UTI) treated with pefloxacin, which is a fluoro-quinolone derivative, in a dose of 800 mg per day. Steroids were avoided because of associated UTI. The UTI responded well and proteinuria disappeared after ten days of treatment with pefloxacin. However, the patient developed arthralgia involving the ankles, the knees and the neck. At this juncture, the drug was discontinued resulting in complete cessation of the joint pain. Pefloxacin increases the production of interleukin-2, a cytokine whose metabolism is modified during nephrotic syndrome. It has been used earlier in children with INS with equivocal results. The toxicity of quinolones for the joints seems more frequent in children, whose cartilage is immature and several cases have been reported. Studies on a larger number of patients are required before drawing any firm conclusions on the usefulness of pefloxacin in the treatment of INS. PMID- 18417915 TI - Renal involvement in henoch-schonlein purpura. PMID- 18417916 TI - Pneumocystis carinii Pneumonia after Renal Transplantation. PMID- 18417917 TI - Kidney transplantation from living unrelated donors. PMID- 18417918 TI - Use of Erythropoeitin in Pre-dialysis Patients. PMID- 18417919 TI - Incidence of ESRD: Magnitude of the Problems and Implications. PMID- 18417920 TI - I want a Saudi Doctor! PMID- 18417921 TI - Establishment of a cadaveric transplantation program: the saudi arabian experience. PMID- 18417923 TI - Living organ donation: time for a donor charter. PMID- 18417922 TI - Islamic ethics of organ transplantation and brain death. AB - The Kingdom of Saudi Arabia, the most conservative Islamic country, is playing a major role in formulating ethical Islamic jurisprudence views and rules in the rapidly expanding field of organ transplantation, and in implementing these new rules in its proliferating centers. The Islamic O' Sool or fundamentals of Islamic jurisprudence through which the jurists reach their rulings are discussed briefly. The historical background of the Islamic jurists' views on organ transplantation and the recent Fatwas (decrees) are highlighted. The conditions needed for allowing both cadaveric and living donors are discussed. PMID- 18417924 TI - What it Means to Die in Islam and Modern Medicine. AB - The objectives of this article is to evaluate whether the concept of brain-death can be examined by modern medicine alone as well as examine the impact of such an evaluation on Islamic views of this subject. Various clinical, philosophical, Islamic, and public-policy literature on the subject of death and brain-death have been examined. We present arguments to support the view that the subject of death (including brain-death) cannot be examined without a philosophical base. Any discussion on death should consist of definition of what it means to die, criteria for determining that death has occurred, and specific medical tests that show whether these criteria have been fulfilled. Medicine has no definition for death based on experimental sciences and death is defined by a philosophical concept. In order to accept the concept of brain-death in Western countries, they had to change first the philosophical definition of death. Also, there is still a debate in modern medicine whether death is an event or a process. Most recent Islamic literature has accepted the concept of brain-death as a medical fact, without discussing its philosophical base. This philosophical definition depends on many subjective factors. In Islam, death has a clear definition: it is the departure of the soul and hence, it is an event. However, the signs of this departure have not been specified and they were left to experts (physicians) to define them. In conclusion, medicine alone cannot formulate a concept regarding death. A philosophical definition of death must be used with it. The Islamic discussion on the concept of death should be focused mainly on its philosophical definition. The definition of death in the concept of "brain-death" does not contradict the concept of death in Islam. PMID- 18417925 TI - Update of Organ Transplantation in the Gulf Co-operation Council Countries. PMID- 18417926 TI - Survey of opinion of secondary school students on organ donation. AB - We conducted a survey of opinion of a sample of senior high school students in Saudi Arabia to evaluate their awareness of the importance of organ donation and concept of brain death. There were 839 students from nine schools, 745 males and 94 females. The participants were not primed about these topics before answering the questionnaire, which was answered at school. The study group declared the level of education of the parents. Twenty three percent knew about the Saudi Center for Organ Transplantation, but 61% could only guess its function. Fifty eight percent could not differentiate between "natural 11 death and brain death and 93% were not aware of how to document it. Elaboration on these questions showed variable explanations. Sixty eight percent agreed to donate organs of relatives in case of brain death, and 91% would donate a kidney to their relatives. However, 38% agreed to donate organs of their own to organ failure patients other then relatives. Thirty five percent knew about the organ donation cards, but only 12% carried them, and only 48% would consent to include the word "donor" on their driving licenses. Forty two percent knew about the opinion of Islamic religion toward organ donation. Thirty one percent agreed to send patients for organ transplantation abroad due to their belief that transplantation technology in Saudi Arabia is lacking, There were no significant differences in the answers according to schools, gender, students of different curricula, having a friend or relative with organ failure, or the level of the education of the parents. This study suggests the great need for education of the new generation about the importance of organ donation and the concept of brain death. We believe that including these topics in the curricula of schools would help disseminating this knowledge to the public in Saudi Arabia. PMID- 18417927 TI - Knowledge and Attitude towards Organ Donation among Males in Riyadh, Saudi Arabia. AB - Organ transplant programs are increasing in Saudi Arabia with the major barrier to transplantation being a shortage of organs. The majority of Saudi Nationals are reluctant and unwilling to donate or consent for donation. This study was undertaken to determine the knowledge and attitude towards organ donation among males in Riyadh, Saudi Arabia. A questionnaire was distributed to 223 men attending the out-patient department of the National Guard Hospital, Riyadh. A total of 205 (92%) individuals answered the questionnaire. Of them, 187 (91%) were Saudis and 18 (9%) were non-Saudis. A total of 187 (88%) had heard about organ donation of whom 80 (43%) each, had acquired this knowledge through television or radio, 16 (8%) through newspaper and magazines, seven (4%) through friends and relatives, and four (2%) through health-care workers. Of the 205 study subjects, 88 (43%) claimed they understood the concept of brain-death, 96 (47%) did not, and 19 (10%) did not respond to this question. One hundred and thirty-eight (67%) were willing to donate, and 156 (76%) were willing to receive an organ. One hundred and fifteen (56%) believed that Islam permits people to donate organs, five (2%) thought Islam does not permit organ donation, 64 (31%) gave a "don't know" answer and 21 (11%) did not attempt to answer the question. In addition, 41 (20%) thought organ donation disfigures the body. In conclusion although 67% of the respondents in this survey were willing to donate, there was a significant lack of knowledge and misconception with regard to Islamic support to, and the mutilating effects of, organ donation. Public educational programs and other measures addressing these issues may help in increasing the rate of organ donation among Saudis. PMID- 18417928 TI - Some of the Lessons Learnt from Renal Transplant Recipients Cared-for at the Riyadh Armed Forces Hospital. AB - The first-ever renal transplantation in Saudi Arabia was carried out at the Riyadh Armed Forces Hospital in March 1979. Since then, 480 renal transplants have been performed in our institution and we have also followed-up many patients who have been transplanted outside the Kingdom. Over 85% of our patients are on cyclosporin-based immunosuppression and the mean follow-up was 3.4 years. In this paper, we summarize our experiences and findings in the field of renal transplantations. PMID- 18417929 TI - Initiating a new renal transplant program: problems and results. AB - Patients from the North-West area of Saudi Arabia had to travel to Riyadh or Jeddah for renal transplantation and post-transplant follow-up. The administration of the North-West Armed Forces Hospitals Program provided medical expertise and financial support to set up a renal transplant program in this area. Suitable staff members including nephrologists, transplant surgeons, staff nurses social workers and laboratory personnel were recruited. A strong link was established with the Saudi Canter for Organ Transplantation in Riyadh. At the end of two years since establishment, 25 renal transplants (11 cadaver donor and 14 living donor) have been performed. During follow-up, nine patients developed serious infection including pneumonia, wound infection and tubercu-losis; all were managed successfully. Three grafts were lost necessitating graft nephrectomy and there were four deaths. The two-year graft survival is 83.3% and patient survival, 84%. These results are highly satisfactory taking into consideration the fact that our transplant program is new. PMID- 18417930 TI - Experience with renal transplantation in the eastern province of saudi arabia. AB - We evaluated our experience with renal transplantation to determine its safety and efficacy. Of the 47 renal transplants performed at our institution from June 1992 to December 1995, 29 were performed from living related donors and 18 from cadaveric donors. The age of the patients ranged from 17 to 65 years with a mean age of 32.2 years. Patient survival, allograft survival and the incidence of complications were used as determinants of successful outcome. Patient survival was 100% for both groups. The allograft survival was 100% for living related donor recipients, with 90% of the cases having normal graft function and 96% for cadaveric recipients with 78% of the cases having normal graft function after a mean follow-up of 24 months (range 3 to 42 months). The incidence of complications encountered was similar to that reported from European and North American renal transplant centers. Our study shows the safety and efficacy of the renal transplantation program in the Eastern Province of Saudi Arabia. PMID- 18417931 TI - Renal transplantation at the jeddah kidney center. AB - Renal transplantation is the optimal treatment for patients with end-stage renal failure. During the period 1991 to 1995, a total of 279 renal transplantations were performed at the Jeddah Kidney Center. They included 115 kidneys from cadaveric donors and 164 living related donor transplants. There were 160 males and 119 females; age of the patients ranged between 4 and 45 years. During the follow-up period, 32 grafts were lost and 26 patients died. The overall 5-year graft and patient survival rates were 79.2% and 90.7% respectively. Sepsis and pulmonary embolism constituted the common causes of death. PMID- 18417932 TI - Renal transplant experience at king fahad national guard hospital. AB - Between September 1988 and end of 1995, 170 renal transplantations were performed at the King Fahad National Guard Hospital, Riyadh. There were 102 cadaver donor (CAD) and 68 living related donor (LRD) transplants. All patients received triple drug immunosuppression. Patients with delayed graft function received anti thymocyte globulin instead of cyclosporin until recovery of graft function. The 1,3 and 5 year graft survival was as follows: all transplants 83.5%, 77.6%, 74.7% respectively; LRD transplants, 89.7%, 85.2%, 80.9% respectively; CAD transplants, 81.0%, 72.5%, 70.5% respectively. The commonest cause of graft loss was chronic rejection. The 1,3 and 5 year patient survival was as follows: all transplants, 97.0%, 94.7%, 93.5% respectively; LRD transplants, 98.5%, 97.0%, 95.5% respectively; CAD transplants, 96.0%, 93.0%, 92.0% respectively. The overall results in our center are comparable to those published from other centers in Saudi Arabia and abroad. PMID- 18417933 TI - The southern region renal transplant program at armed forces hospital, khamis mushayt. AB - The Southern Region renal transplant program was established in February 1989. The appointment of a transplant co-ordinator and creation of a waiting list for the Southern Region as well as tissue typing of all patients in the region were important early steps. Between February 1989 and December 1995, 155 transplants were performed on 152 patients at the Armed Forces Hospital, Southern Region (AFHSR). Of them, 52 were cadaveric donor transplants and the remaining were from living related donors. The overall five-year actuarial patient and graft survival was 93% and 78% respectively. Of the 152 patients who were transplanted, 79 patients were from other hospitals in the region and 73 were from AFHSR. Maintenance immunosuppression consisted of cyclosporin, azathioprine and prednisolone. Use of the spouse as a donor was an early feature of this program. Our results compare favorably with results published from other centers. To cope with the increasing demand of transplantation in the Southern Region, we have to look into ways of increasing our transplant numbers to match the needs. PMID- 18417934 TI - Complications in kidney transplant recipients: a single center experience. AB - A total of 147 kidney transplant recipients are on regular follow-up in our center including 12 who were transplanted in our hospital since the start of the transplantation program in June 1994. Fifty eight patients received cadaveric kidneys (mainly in the United States), 13 received living-related transplants (of which seven were performed in our hospital), and 76, received living non-related transplants abroad. The follow-up period ranged from 0-228 months. Fifty eight (39.5%) patients continue to be followed with functioning kidneys, 26 (17.7%) patients lost their grafts, 19 (12.9%) patients died, and 44 (29.9%) were lost to follow-up. Six patients converted to HIV-positive after transplantation, of which five had received living non-related transplantation. Four of these patients died. Other significant infectious complications during follow-up period included pneumonia (n = 37; 25%) and tuberculosis (n = 9; 6%). Hypertension was present in 86% and diabetes mellitus in 30% of the patients. Hepatitis developed in 24% of the patients. Eight patients had pregnancy followed by successful delivery (two of them twice). Twenty seven patients had significant post-operative urological complications, including 12 lymphoceles and three urinary leaks. When the influence of different factors such as type and place of transplant, immunosuppression, infections, and occurrence of acute rejections, hypertension, diabetes, hepatitis and urological complications, on outcome were analyzed, only history of acute rejections and chest infections had a significant independent effect. PMID- 18417935 TI - Liver transplantation at king fahad national guard hospital riyadh, kingdom of saudi arabia. AB - Liver disease is a major medical problem in the Kingdom of Saudi Arabia and is mostly due to viral hepatitis. Liver transplantation is the only option for patients with end-stage liver disease offering good long-term survival. The first liver transplant at the King Fahad National Guard Hospital was performed in February 1994 and since then, 40 liver transplants have been performed on 37 patients. Immunosuppression consisted of prednisone combined with cyclosporin (Neoral) or FK 506. Maintenance immunosuppression was with the use of cyclosporin or FK 506 as monotherapy. All, but one patient, survived the surgical procedure; there were no cases of primary non-function; acute cellular rejection occurred in 12 patients all of whom responded to steroids. Pneumonia and biliary sepsis occurred in 12 patients each. A total of 10 patients died, with sepsis being the leading cause of death. The overall graft survival was 73%. Donor shortage continues to be a major limiting factor. PMID- 18417936 TI - Story of the first liver transplant in saudi arabia. AB - Chronic liver disease is common in Saudi Arabia with viral hepatitis and schistosomiasis being the common causes. I herewith describe the experience on the first liver transplantation in Saudi Arabia and the Arab World. The patient was a 23-year old Saudi male who had cirrhosis of the liver secondary to sclerosing cholangitis. He underwent orthotopic liver transplantation on July 30, 1990. The post-operative course was uneventful and immunosuppression comprised of small doses of cyclosporine, azathioprine and prednisolone. The patient continues to be well six years post-transplant and is adequately rehabilitated. PMID- 18417937 TI - Orthotopic liver transplantation at king faisal specialist hospital and research center: 1994-1995. AB - Over the last decade, orthotopic liver transplantation (OLT) has become an established therapy for end-stage liver disease of various etiologies. The early experience with orthotopic liver transplantation in the Kingdom was in 1990 in the Military Hospital when a man with sclerosing cholangitis received a new liver successfully. Intensive effort was done at King Faisal Specialist Hospital and Research Center (KFSH & RC) to start liver transplantation and that was achieved in March 1994 when a man with an end-stage liver failure secondary to hepatitis C was transplanted successfully. Since then, forty four (44) more liver transplantations were done at KFSH & RC. The age of the patients transplanted ranged from 9-65 years old; there were more males than females, (26 males, 15 females). The waiting time until transplantation was up to one year. All patients received a combination of cyclosporin and prednisolone as an induction therapy +/ Azathioprime. The majority of patients developed minor complications like wound infection and acute mild cellular rejection. In the second year, 3 out of 18 patients also developed primary non-function. Also in the first year, the majority of the patients developed primary dysfunction; however, this decreased significantly in the second year. The majority of the patients who were transplanted for hepatitis C had mild recurrence. None of them lost their livers because of recurrence of hepatitis C. Several patients developed biliary complications including bile leak and stricture at duct-to-duct anastomosis. PMID- 18417938 TI - Orthotopic heart transplantation in the prince sultan cardiac center. AB - In this report we attempt to demonstrate the efforts involved in establishing and organizing the heart transplant program at the Armed Forces Hospital in Riyadh, Saudi Arabia. From 1986 to date, 25 orthotopic heart transplants were performed at this center. Patient age ranged from 22 months to 57 years; 4 patients were below 12 years of age and 4 aged 50 years and above. The incidations for transplantation were cardiomyopathy in 15 patients, ischemic heart disease in 6 patients, and valvular heart disease in 4 patients. Fourteen recipients have died. Three of them were classified as hospital deaths, occuring before the patient could be discharged after the procedure; the reminder died from rejection and associated problems. Eight patients of them died within the first year. The longest survival period was almost 8 years. The overall 8 years survival rate was 45%, which is comparable to the international figures. Shortage of donors may affect the future of the transplant programs. Increasing the awareness of the public about the importance of organ donation and transplantation is crucial in this regard. PMID- 18417939 TI - Single lung transplantation for alveolar micro-lithiasis: the first clinical report. AB - Pulmonary alveolar microlithiasis (P.A.M.) is a rare pulmonary disorder that pursues usually an asymptomatic course and can culminate in severe respiratory failure. We report a 48 year old Saudi female patient with P.A.M. who deteriorated rather steadily after the initial 18 years of asymptomatic course until a frank type I respiratory failure is established. Single lung transplantation (S.L.T.) was performed successfully and the patient returned to full daily activity and has now survived 12 months post S.L.T. The immunosuppression consisted of Cyclosporine-A 10 mg/kg/day, azathioprine (immuran) 2 mg/kg/day and prednisolone 10 mg daily. The bronchial anastomosis was done by telescoping the recipient and donor main bronchus without omental wrap. A significant bronchial stricture of the anastomotic site occurred 4 months post S.L.T. which was dilated endoscopically with good clinical and bronchoscopic result. No episodes of rejection or infection were encountered so far. PMID- 18417940 TI - Bone marrow transplantation in children: the king faisal specialist hospital experience. AB - The results of the Pediatric Bone Marrow Transplant Program at The King Faisal Specialist Hospital and Research Center (KFSH & RC) from June 1993 to October 1995 were reviewed for a preliminary report on the outcome of children undergoing bone marrow transplantation (BMT) particularly in relation to mortality and morbidity. A total of 64 bone marrow transplants were performed on 60 patients during this period of time. The study patients included 28 with acute leukemia, 10 with severe combined immune deficiency, five with chronic myeloid leukemia, four with Fanconi's anemia and 13 others with miscellaneous disorders. The average hospitalization period was six weeks per patient. Forty three of these patients (72%) were alive and disease-free after a median follow-up of 14 months (range 1-27 months). Eight patients died from transplant-related toxicity within 100 days of BMT. One patient died of chronic graft versus host disease (GVHD) of the liver. Eight patients with acute leukemia relapsed within one year after BMT. Further details regarding the preparative regimens, toxicity of BMT, GVHD and disease-free survival are reviewed in this report. PMID- 18417941 TI - The experience of bone marrow transplantation at riyadh armed forces hospital. AB - This is a report about the program of bone marrow transplantation (BMT), which was established in 1989 at the Armed Forces Hospital in Riyadh, Saudi Arabia. We follow the strict international protocol of pre transplant assessment of the donor and the recipient, BMT conditioning by Cyclosphosphamide and Basulphan or body irradiation, BM harvest and processing, graft versus host disease prophylaxis by cyclosporin and methotrexate, and post-transplant care. Since the start of the BMT programme at the Armed Forces Hospitals in Riyadh in May 1989 and until the end of March 1996, fifty nine allogeneic and one autologous transplants have been performed. Chronic myelocytic and acute myeloid leukemia were the principal indications for BMT in our institute. The acturial five years survival of BMT in these two conditions was 67% and 47% respectively. Besides allografting, we finished the preparations for autologus BMT and peripheral blood stem cells transplantation, which may be used for treating patients with solid tumors and leukemias who are not suitable for allogenic transplantation due to older age or donor unavailability. PMID- 18417942 TI - The role of women in consenting for organ donation in saudi arabia. PMID- 18417943 TI - Hepatorenal syndrome. AB - The hepatorenal syndrome (HRS) is a common complication of advanced liver disease. Careful management can reduce the risk of this functional renal failure developing in the vulnerable group of patients. Potentially nephrotoxic agents (drugs, x-ray contrast, etc.) need to be avoided wherever possible. It is likely that sympathetic overactivity, endotoxin, nitric oxide and endothelin, together with perturbed arachidonic acid metabolism, are closely involved in pathogenesis. Many of the associated abnormalities in HRS can be explained on the basis of an inadequate effective arterial blood volume and the resulting attempts at compensatory homeostasis. Medical management is disappointing, but trans cutaneous intra-hepatic portal systemic shunting may help. Successful orthotopic liver transplantation is curative. PMID- 18417944 TI - Blood pressure among school children in jordan. AB - A prospective study was carried out over a three-year period (1993-95) on 4469 school children drawn from 20 different schools in rural areas of Jordan. There were 2592 males and 1877 females aged between 6-16 years. The height, weight, systolic and diastolic blood pressure (BP) were recorded for both sexes followed by complete clinical examination. The BP was recorded according to the criteria laid down by the second task force on BP in children, using mercury sphygmomanometers, in sitting position and in the right arm. Data were analyzed and the percentiles were calculated for each age-group in both sexes. Both systolic and diastolic BP had positive correlation with age, height, weight and body surface area. There were no differences in the systolic BP for both sexes of corresponding age, while there was a difference in the diastolic. The upper limits of normal, 90th percentile, systolic/diastolic pressures were 116/76, 122/80, 128/81 and 139/86 in children aged 6-8 years, 9-11 years, 12-14, and 15 16 years respectively, with prevalence of 13.35% (n = 596). The lower limits of hypertension, 95th percentile, for systolic/diastolic pressures were 122/81, 126/83, 134/84, and 142/88 mm Hg in each of the same age-groups respectively, with prevalence of 6.85% ( = 306), while for severe hypertension, 99th percentile, for the same age-groups the values were 131/86, 134/87, 145/89 and 154/90 mm Hg respectively, with prevalence of 1.95% (n = 87). The findings in this study were consistent with international data. We emphasize the need for regular check up of BP in our children. Also, further studies are necessary including other areas of Jordan and smaller age-group children. PMID- 18417945 TI - Is there an advantage in adding azathioprine to prednisone and cyclosporine in kidney transplant recipients? AB - Some recent studies have raised doubts about the advantage of adding azathioprine to cyclosporine and prednisone ("triple therapy") in kidney transplant patients. We reviewed the charts of 53 patients followed at our center after they received a living non-related kidney transplant in India. Thirty-eight patients used "triple therapy" (cyclosporine, prednisone and azathioprine) and 15 patients "double therapy" (cyclosporine and prednisone). A multi-variate analysis was used to study the differences in gender, age, follow-up period, number of patients with at least one acute rejection episode, number of significant infections, cyclosporine dosage and trough levels, prednisone dosage, transplant function, and graft loss due to rejection. There was no difference in the number of acute rejections or graft loss due to rejection in both treatment groups. Although the mean cyclosporine levels at 6, 12 and 24 months and the mean dosage at 6 and 12 months were significantly lower in the triple therapy group, there was no significant differences in these parameters between the patients with and without rejection. The cyclosporine levels at the time of rejection were also not different from the levels in the patients without rejection. The results support other reports suggesting that azathioprine does not seem to add to the immunosuppressive effect, as it is not associated with a lower incidence of acute rejections or an improved graft survival. PMID- 18417946 TI - Thyroid function in children with chronic renal failure. AB - This study was undertaken to test the thyroid function in non-dialyzed children with chronic renal failure (CRF). Sixteen children with CRF and 12 healthy children who served as controls were studied for their thyroid function status. We found a significant increase in the serum thyroid stimulating hormone (TSH) level and a significant decrease in serum Triiodo thyronine (T3) levels in children with CRF as compared to the healthy children (p 0.2). There was a weak negative correlation between hemoglobin concentration and serum TSH (r = - 45) as well as between blood urea nitrogen and serum T3 (r= -0.30) in children with CRF. These results suggest that children with CRF commonly have a state of biochemical hypothyroidism although they are clinically euthyroid. Early renal transplantation is indicated to correct such metabolic defects. PMID- 18417947 TI - Vesicoureteral reflux in children. Experience in riyadh, saudi arabia. AB - There are a few reports about the size of the problem of vesicoureteral reflux (VUR) in developing countries. We attempted in our study to assess the experience of this problem in children in a tertiary care medical center in the period between June 1983 till June 1993. VUR was diagnosed in 24 patients, of whom 71% were boys. The mean age of the patients was 36.5 months; seven (29%) of them were below one year of age. The commonest presentation was urinary tract infection. E. coli was the most common organism and was resistant to the first chpice chemoprophylaxis in 50% of cases. Urine culture was also positive in 37.5% of children who were grossly asymptomatic. Ultrasound study failed to detect VUR in 25% of cases. There were eleven patients with mild to moderate reflux (grades I, II, III) of whom 9% required ureter reimplantation by open surgery. Eleven patients had severe reflux (grades IV,V) of whom 72% required the same procedure. Post-operatively, of the nine patients who had reimplantation, seven (77.5%) had successful surgery and maintained normal renal function after a mean duration of follow up of 30 months (range from 6 to 84 months), one patient developed end stage renal disease, and one patient was lost to follow-up. We conclude that VUR is not uncommon in early childhood. Early referral of severe cases for surgical reimplantation is recommended. PMID- 18417948 TI - Sarcomatoid renal cell carcinoma associated with renal schistosomiasis: a case report and review of literature. AB - A 77 year old Saudi male admitted to the surgical unit of our hospital because of a left renal mass. A diagnosis of left renal carcinoma was subsequently made and a left nephrectomy was performed. Histological examination of the nephrectomy specimen showed a renal cell carcinoma containing a large sarcomatoid component. In addition, numerous schistosoma ova, some of them calcified, were identified within the carcinomatous part of the tumor. To the best of knowledge, this is the first report of simultaneous renal schistosomiasis and sarcbmatoid renal cell carcinoma. PMID- 18417949 TI - Acute renal failure following snake bite: case report and review. AB - We present an adult male patient who presented with history of having been bitten by a snake. Despite administration of antivenom and adequate hydration, he developed acute renal failure secondary to disseminated intravas-cular coagulation and rhabdomyolysis. He improved with daily hemodialysis and at discharge had normal renal function. Although we did not perform a renal biopsy on him, the clinical course was highly suggestive of acute tubular necrosis. The pathogenesis, pathology, as well as therapy of renal involvement following venomous snake bite is discussed. PMID- 18417950 TI - Renal replacement therapy in yemen. AB - In this report we present the current status of dialysis and transplantation in Yemen. The reported incidence of end stage renal disease (ESRD) in one region of Yemen was estimated as 385 per million population (PMP) per year. The total population of Yemen is also estimated as 16,000,000. Peritoneal dialysis was started in 1980, while hemodialysis was started in 1981. At present there are around 36 hemodialysis machines distributed in the large cities of Yemen. Intermittent peritoneal dialysis is commonly used; however, continuous ambulatory peritoneal dialysis has been out of practice since 1992. Renal transplantation has not yet been started in Yemen; however, at present there are 327 transplant patients being followed up in it. The majority of patients had their grafts from living non related donors abroad. In our experience, such transplantations were associated with high morbidity and mortality, in addition to acquisition of serious, potentially lethal extra-renal medical problems. We believe that there is a wide shortage of renal services in Yemen. Establishing a National Kidney Foundation to organize these services may be helpful. PMID- 18417951 TI - Cyclosporine a induced hyperkalemia in a renal allograft recipient. PMID- 18417952 TI - How long should a patient with kidney transplant continue on immunosuppressive drugs ? PMID- 18417953 TI - Adding color and flavor. PMID- 18417954 TI - Home non-invasive mechanical ventilation and long-term oxygen therapy in stable hypercapnic chronic obstructive pulmonary disease patients: comparison of costs. AB - BACKGROUND: A cost analysis of nocturnal non-invasive ventilation (NNV) in stable chronic obstructive pulmonary disease (COPD) patients would be helpful in decision making, when the balance between the increased demand and the availability of resources should be checked. OBJECTIVES: Based on data from the Italian trial in stable hypercapnic COPD patients, this study compares the cost of care associated with the use of NNV when added to the usual long-term oxygen therapy (LTOT) with the cost of care of LTOT regimen alone. METHODS: Cost was calculated in 77 of 90 patients included into that trial. Analysis included drug therapy, hospitalisations due to acute exacerbation, oxygen and ventilator equipment. An estimation of charges was made according to the national sources of cost for drugs and hospital admissions and the actualised reimbursement for the home care provided to both oxygen and ventilator users. The cost/day comparison was made between the individual patients in the 2 groups (NNV + LTOT, n = 35; LTOT, n = 42). RESULTS: The mean cost of drugs and oxygen was similar in both groups, whereas the cost of hospitalisation tended to be lower in NNV + LTOT compared to LTOT alone (8.25 +/-10.29 vs. 12.50 +/- 20.28 EUR/patient/day, p < 0.05). Inclusion of the ventilator equipment increased the total cost to 23.73 EUR/day in the NNV + LTOT compared to 21.42 EUR/day in the LTOT group (not significant). CONCLUSIONS: The present report suggests that long-term management with addition of non-invasive ventilation does not increase costs compared with the usual LTOT regimen: the hospital-related costs were reduced when using the ventilator in these hypercapnic COPD patients. PMID- 18417955 TI - Markedly different clinical features in 2 diabetes mellitus patients with extremely high tissue levels of the mitochondrial DNA A3243G mutation. AB - BACKGROUND: Mitochondrial DNA (mtDNA) A3243G mutation is one of the major causative factors of mitochondrial diabetes mellitus. We found that tissues from 2 of 142 diabetes mellitus patients showed extremely high levels of the mutation. OBJECTIVE: To investigate the level of the mutation in each tissue and to find the relationship between the mutation level and clinical features of the patients. METHODS: Patient 1 was a 51-year-old woman, diagnosed as having diabetes mellitus at the age of 17, and was admitted to hospital because of cerebral infarction. Patient 2 was an 82-year-old woman who was admitted because of respiratory failure. mtDNA A3243G levels were measured in tissues collected at autopsy. RESULTS: In patient 1, mtDNA A3243G levels were found to vary among the tissues. The patient's highest mtDNA A3243G value was 42% and the lowest value was 9%, whereas the level in most individuals is usually less than 1%. Although patient 2 did not exhibit serious clinical symptoms of diabetes mellitus, the mtDNA A3243G level was extremely high in all of the tissues surveyed (range 32 47%). CONCLUSION: Although both patients showed high levels of the mtDNA A3243G mutation, their clinical conditions differed greatly. Thus, mitochondrial diabetes mellitus patients may show a wide variety of clinical features and large variations in life span. PMID- 18417956 TI - Lymphoma and leukemia cells possess fractal dimensions that correlate with their biological features. AB - BACKGROUND: Living cells can be viewed as complex adaptive systems that exhibit non-linear dynamics and fractal features. We investigated the fractal qualities of normal and malignant hematological cells and their potential as a tool for characterizing cell phenotype and clinical behavior. METHODS: A mathematical algorithm and an optic tool for fractal analysis of nuclei were developed. A total of 4,713 lymphoid cells derived from 66 patients of five distinct diagnostic groups (normal and reactive lymphocytes, low-grade lymphomas and an aggressive lymphoma) were assessed for their fractal dimension. In addition, in 19 patients fractal analysis of leukemia cells was compared to clinical endpoints. RESULTS: After validating our method, hematological cells possessed fractal dimensions corresponding to their clinical entity. There was a highly significant overall difference in fractal dimensions between various types of hematological malignancies. A preliminary correlation was found between the fractal dimension and the clinical outcome of leukemia patients. CONCLUSIONS: Hematological cells possess fractal dimensions that correlate with their biological properties. Measurement of fractal dimension seems to be a sensitive method to assess the hematological cell phenotype and to define a clinical group. This tool may be potentially useful for the evaluation of clinical behavior of hematological diseases. PMID- 18417957 TI - Peroxisome proliferator-activated receptor-gamma gene polymorphism and risk of cardiovascular disease in patients with diabetic nephropathy. AB - BACKGROUND: Peroxisome proliferator-activator receptor-gamma (PPAR-gamma) is a nuclear receptor that serves important roles in intermediate metabolism. We examined the relationship between two PPAR-gamma gene polymorphisms, namely the P12A and C161T, and cardiovascular disease in patients with diabetic nephropathy. METHODS: We studied 170 predialysis and 50 peritoneal dialysis patients with diabetic nephropathy. The PPAR-gamma genotypes were determined by polymerase chain reaction-restriction fragment length polymorphism assay. The patients were then followed prospectively for the development of cardiovascular events. Cardiovascular mortality and all-cause mortality were also compared. RESULTS: There were 91 male cases. The mean age was 64.3 +/- 10.6 years. The frequencies of Pro/Pro and Pro/Ala genotypes of the P12A polymorphism were 95 and 5%, respectively; CC, CT and TT genotypes of the C161T polymorphism were 55.5, 39.5 and 5.0%, respectively. For the P12A polymorphism, the event-free survival was 56.5 and 9.1% at 60 months for Pro/Pro and Pro/Ala genotypes, respectively (p = 0.0005). For the C161T polymorphism, the event-free survival was 61.5 and 44.9% for CC and CT/TT genotypes, respectively (p = 0.0044). By multivariate analysis with the Cox proportional hazard model to adjust for confounders, the independent factors for event-free survival were P12A and C161T polymorphisms, age and diastolic blood pressure. In this model, the Pro/Ala genotype conferred 7.6-fold (95% CI 2.1- to 28.0-fold, p = 0.002) excess hazard of developing primary cardiovascular end point as compared to the Pro/Pro genotype, while each T allele at the 161 position conferred 83.4% (95% CI 15.2-291.9%, p = 0.011) excess hazard. CONCLUSIONS: We conclude that P12A and C161T polymorphisms of the PPAR gamma gene are important predictors of cardiovascular event in patients with diabetic nephropathy. Further studies are needed to examine the interaction of PPAR-gamma gene polymorphisms and thiazolidinedione treatment on the cardiovascular risk in this group of patients. PMID- 18417958 TI - Effect of carnitine deprivation on carnitine homeostasis and energy metabolism in mice with systemic carnitine deficiency. AB - BACKGROUND/AIMS: Juvenile visceral steatosis (jvs-/-) mice lack the activity of the carnitine transporter OCTN2 and are dependent on carnitine substitution. The effects of carnitine deprivation on carnitine homeostasis and energy metabolism are not known in jvs-/- mice. METHODS: jvs-/- mice were studied 3, 6 and 10 days after carnitine deprivation, and compared to jvs-/- mice substituted with carnitine, wild-type (jvs+/+) and jvs+/- mice. Carnitine concentrations were assessed radioenzymatically. RESULTS: Compared to wild-type mice, carnitine treated jvs-/- mice had decreased plasma beta-hydroxybutyrate levels and showed hepatic fat accumulation. The carnitine levels in plasma, liver and skeletal muscle were decreased by 58, 16 and 17%, respectively. After ten days of carnitine deprivation, the plasma carnitine concentration had fallen by 87% (to 2.3 mumol/l) and the tissue carnitine levels by approximately 50% compared to carnitine-treated jvs-/- mice. Carnitine deprivation was associated with a further drop in plasma beta-hydroxybutyrate and increased hepatic fat. Skeletal muscle glycogen stores decreased and lactate levels increased with carnitine deprivation, whereas tissue ATP levels were maintained. CONCLUSIONS: In jvs-/- mice, tissue carnitine stores are more resistant than carnitine plasma concentrations to carnitine deprivation. Metabolic changes (liver steatosis and loss of muscle glycogen stores) appear also early after carnitine deprivation. PMID- 18417959 TI - Altered erythrocyte membrane protein composition in chronic kidney disease stage 5 patients under haemodialysis and recombinant human erythropoietin therapy. AB - Our aim was to evaluate red blood cell (RBC) membrane protein composition in chronic kidney disease (CKD) stage 5 patients under haemodialysis (HD) and recombinant human erythropoietin (rhEPO) therapy, and its linkage to rhEPO hyporesponsiveness. We evaluated in 63 CKD stage 5 patients (32 responders and 31 non-responders to rhEPO therapy) and in 26 healthy controls RBC count, haematocrit, haemoglobin concentration, haematimetric indices, reticulocyte count, reticulocyte production index, RBC osmotic fragility test and membrane protein analyses. CKD stage 5 patients presented significant changes in membrane protein composition, namely a reduction in spectrin, associated to altered protein 4.1/spectrin and spectrin/band 3 ratios. Non-responder CKD stage 5 patients were more anaemic, with more microcytic and anisocytic RBCs, than responders; significantly altered ankyrin/band 3 and spectrin/ankyrin ratios were also observed. CKD stage 5 patients under HD are associated with an altered protein membrane structure, which seems to the disease itself and/or to the interaction with HD membranes. PMID- 18417960 TI - Peritoneal dialysis in a patient with neurogenic bladder and chronic kidney disease with ventriculoperitoneal shunt. AB - Long-term dialysis in children with multiple handicaps has become easier with the advent of continuous ambulatory peritoneal dialysis (PD). Due to the widespread use of PD and the long survival of patients with spina bifida, an increasing number of patients with spina bifida are on PD. The viability and safety of PD in spina bifida patients with a ventriculoperitoneal shunt (VPS) have been a matter of concern. Some authors consider the presence of a VPS a relative contraindication for PD, but more recent reports suggest that PD under close monitoring is not contraindicated. We report a 17-year-old girl born with meningomyelocele, hydrocephalus and neurogenic bladder who was maintained on VPS. She reached end-stage renal failure 17 years later and was put on PD based on family and patient preference. She had an uneventful course in the initial 9 months, but later developed fungal peritonitis which was successfully managed with catheter withdrawal and an intravenous antifungal agent (amphotercin 0.75 mg/kg). Simultaneous ventricle-aspirated cerebrospinal fluid was sterile. To our knowledge, this is the first report of fungal infection in such a patient. Although we share the view that PD is not an absolute contraindication in patients with a functioning VPS, its likely complications, especially infectious complications in developing countries, should be kept in mind before initiating PD in such patients. PMID- 18417961 TI - Quality of life in stroke survivors after local intra-arterial thrombolysis. AB - BACKGROUND: We aimed to assess quality of life (QOL) and its predictors in stroke survivors after local intra-arterial thrombolysis (IAT) as well as to measure QOL according to the site of pretreatment vessel occlusion. METHODS: From January 2000 to April 2004, 175 consecutive patients underwent IAT for acute ischemic stroke. Clinical and radiological data were collected prospectively. We contacted 135 stroke survivors after a mean of 923 (+/-431) days, 132 responded. QOL, assessed with EuroQol (EQ-5D), and functional abilities, measured with the modified Rankin Scale (mRS) and the Barthel Index, were compared, and predictors of QOL were analyzed. RESULTS: Measured with EQ-5D, 56% of the patients reported a good QOL (EQ-5D Index >or=70). Low mRS and high Barthel Index scores at follow up were associated with better overall QOL (Kendall's tau >0.5). Nevertheless, 25% of the functionally independent patients (mRS 0-2) indicated a markedly impaired QOL (EQ-5D Index <70) and 10% of disabled patients indicated good QOL. QOL was significantly lower in patients with occlusion of the internal carotid artery compared to patients with occlusion of the basilar artery or the M1, M2 or M3/4 segment of the middle cerebral artery (EQ-5D Index: p = 0.005). A high National Institute of Health Stroke Scale score on admission and occlusion of the internal carotid artery were independent predictors of impaired QOL (p < 0.05). CONCLUSION: More than half of the stroke survivors treated with IAT reported a good QOL, mostly survivors with mild disabilities. QOL assessment gives information that is not provided by traditional outcome scores. Our results support guidelines to measure QOL in stroke research. PMID- 18417962 TI - Glutathione peroxidase 1 C593T polymorphism is associated with lobar intracerebral hemorrhage. AB - BACKGROUND: Oxidative stress plays an important role in vascular pathology and contributes to the pathophysiology of primary intracerebral hemorrhage (PICH). Glutathione peroxidase 1 (GPX1) is a key enzyme of the antioxidant system. Here, we investigated whether a functional C593T polymorphism of GPX1 gene is associated with PICH in a Polish population. METHODS: Genotyping was performed in 192 PICH patients and 197 unrelated controls matched for age and sex. All were of Caucasian origin. The C593T GPX1 polymorphism was investigated using the polymerase chain reaction/restriction fragment length polymorphism method. RESULTS: Multivariable logistic regression analysis revealed a significant association between genotypes containing the T allele and the entire PICH group (OR = 1.53; 95% CI = 1.02-2.29) and lobar PICH (OR = 2.36; 95% CI = 1.31-4.26) but not nonlobar PICH (OR = 1.19; 95% CI = 0.75-1.89). CONCLUSIONS: We found a positive association between the studied GPX1 polymorphism and lobar PICH in a Polish population. PMID- 18417963 TI - Danqi Piantan Jiaonang does not modify hemostasis, hematology, and biochemistry in normal subjects and stroke patients. AB - BACKGROUND AND OBJECTIVE: Previous studies on Danqi Piantan Jiaonang (DPJ, NeuroAid), a traditional Chinese medicine, in stroke patients showed promising results. Our aim was to determine the safety of DPJ in normal subjects and stroke patients through a series of studies assessing its immediate and long-term effects, alone and in combination with aspirin, on hematological, hemostatic, and biochemical parameters. METHODS: We conducted 3 studies from December 2004 to May 2006. Study 1 was a case series which recruited 32 healthy volunteers who were given 2 oral doses of 4 DPJ capsules (0.4 g/capsule) 6 h apart. Study 2 was a randomized controlled trial of 22 healthy volunteers who received either 1 oral dose of aspirin 300 mg alone or a combination of 1 dose of aspirin 300 mg and 2 doses of 4 DPJ capsules taken 6 h apart. For both studies 1 and 2, hemostatic parameters (prothrombin time, activated partial thromboplastin time, fibrinogen, platelet aggregation, D-dimer) were tested at baseline, and after 2 and 8 h. Study 3 was a case series which recruited 10 patients with recent ischemic stroke (within 7 days) who were given 4 DPJ capsules taken orally 3 times a day for 1 month. Blood tests for hemostatic, hematological (complete blood count), and biochemical parameters (glucose, creatinine, alanine aminotransferase, aspartate transaminase, C-reactive protein) were performed at baseline, and after 1 and 4 weeks. RESULTS: Apart from the expected changes in platelet aggregation in subjects taking aspirin, no significant differences were detected in hemostatic parameters at baseline, and 2 and 8 h after oral intake of DPJ alone or in combination with aspirin. Likewise, no significant differences were observed in hematological, hemostatic, and biochemical parameters at baseline, and after 1 and 4 weeks of oral intake of DPJ. CONCLUSION: DPJ does not significantly modify hematological, hemostatic, and biochemical parameters in normal subjects and stroke patients. PMID- 18417964 TI - Special issue: enhancing minority recruitment into genetics research. PMID- 18417965 TI - Increasing minority participant enrollment into a cancer family registry: the Cancer Genetics Network. PMID- 18417966 TI - BRCA1 and BRCA2 risk perceptions among African American women at increased risk for hereditary breast-ovarian cancer. AB - OBJECTIVES: To describe BRCA1 or BRCA2 (BRCA1/2) risk perceptions among African American women at increased risk for hereditary breast-ovarian cancer and to identify factors having independent associations with these perceptions. METHODS: Risk perceptions were evaluated by self-report during a structured telephone interview among African American women (n = 162) at increased risk for hereditary cancer who were recruited from oncology clinics, general medical practices, and community oncology resources. RESULTS: The majority of women (75%) believed that it was likely that they had a BRCA1/2 mutation. Women ages 50 and younger and those with greater cancer-specific worry were most likely to believe that they had a BRCA1/2 mutation. CONCLUSIONS: Although BRCA1/2 risk perceptions may be consistent with objective risk levels among African American women, discussion about the basis of risk perceptions may enhance provision of genetic counseling and testing in this population. PMID- 18417967 TI - Partnership with an African American sorority to enhance participation in cancer genetics research. AB - BACKGROUND/AIMS: Reduced minority participation in clinical research challenges researchers to consider novel recruitment modalities. This study describes a formal partnership between the National Educational Foundation of the Zeta Phi Beta Sorority and the Mid-Atlantic Cancer Genetics Network. The goal was to enhance awareness about inherited breast cancer and to increase enrollment in the national Cancer Genetics Network. METHODS: In this descriptive, pilot study, two recruitment strategies across four states were undertaken: an onsite educational session at four Annual State Leadership Conferences and a 2-tiered direct mail campaign to the sorority membership. RESULTS: Recruitment methods targeted over 1,200 well-educated African American women. Of the 279 attendees at the state conference educational sessions, only 3 women meeting the high risk eligibility requirement enrolled. Direct mail recruitment elicited 24 eligible women. Lessons learned are described. CONCLUSION: Despite low accrual, the partnership laid a foundation for broader collaboration with the Zeta Phi Beta Sorority. In the future, collaboration with minority sororities and fraternities as part of standard registry recruitment should be explored. PMID- 18417968 TI - Successful strategies for increasing African American participation in cancer genetic studies: hopeful signs for equalizing the benefits of genetic medicine. AB - OBJECTIVE: To determine whether prior success in recruiting African Americans to an in-house cancer genetics registry could be duplicated when recruiting to a national registry requiring a significantly increased level of commitment. Additionally, to determine which recruitment sources and practices yielded the highest number of African American participants. METHODS: A retrospective analysis of recruitment sources, practices, and results for recruitment to the Cancer Genetics Network (CGN; a national research registry), from 2000 to 2005 was conducted. These results were compared to previous experience in recruiting African Americans to the Family Cancer Registry (FCR; an in-house registry) during the period 1992-2005. RESULTS: In the 1st year of recruitment to the CGN, African Americans accounted for 24% of those consenting to participate in the CGN registry from our center. This compares to an average annual rate of 27% for the FCR during the years 1998-2005, and a rate of less than 1% from 1992 to 1998. By 2005, African Americans accounted for 27% of CGN participants recruited through the University of Texas Southwestern Medical Center, one of eighteen participating institutions in the CGN. Hospital-based resources such as cancer treatment clinics and tumor registries yielded the highest percentage of African American participants (66.5%), and self-referral yielded the lowest (0%). Seventy seven percent of African Americans were actively sought out and recruited from treatment clinics, whereas the vast majority of Caucasian participants were recruited passively during the course of genetic counseling sessions that were scheduled for reasons unrelated to participation in cancer research. There were no known instances of African Americans contacting CGN staff after reading printed recruitment materials or internet advertisements. CONCLUSIONS: The increased level of commitment required of CGN participants did not deter African Americans from participating in cancer genetics research. Recruitment strategies responsible for dramatically increasing recruitment rates to the FCR from 1998 to 2000 were equally effective when used for recruitment to the CGN. The most effective recruitment sources were high-yield venues such as cancer treatment clinics and tumor registries, and active recruitment methods yielded the highest number of African American participants. Advertising through internet announcements and printed recruitment materials did not appear to be effective. PMID- 18417969 TI - Testing three different cancer genetics registry recruitment methods with Hispanic cancer patients and their family members previously registered in local cancer registries in Texas. AB - OBJECTIVE: To increase accrual among Hispanics to the Cancer Genetics Network national cancer genetics registry. METHODS: Drawing from South Texas cancer registries, 444 Hispanic men and women were randomly assigned to one of three experimental conditions: standard direct-mailed procedures (X1), X1 plus culturally tailored materials (X2), and X2 plus interpersonal phone contact (X3). Participants were also surveyed about the effectiveness of the education materials and the phone contact. A refusal survey was provided for those who declined to join the study. RESULTS: A total of 154 individuals joined the Cancer Genetics Network. The X3 condition yielded the greatest accrual (43.2%) compared to X1 (30.9%) and X2 (29.9%; p < 0.05). Tailored materials appeared to have no effect but were highly regarded. The main reasons for not participating were a lack of interest and time requirements. CONCLUSION: Interpersonal communication can have a powerful effect on recruitment. However, more research is needed to determine the cost-efficacy of more labor-intensive approaches to registry accrual. PMID- 18417970 TI - Factors associated with African Americans' enrollment in a national cancer genetics registry. AB - This study explored whether reactions to the Cancer Genetics Network (CGN) or CGN enrollment differed by receipt of a standard informational brochure versus a targeted version addressing factors previously associated with African Americans' health behavior decisions and research participation. The 262 participants, identified through tumor registries or clinic contacts, were mailed brochures and completed phone interviews. When asked whether - based on the brochure - they were or were not 'leaning toward' CGN enrollment, about 75% of both standard and targeted groups reported leaning toward. When given the opportunity at the end of the interview, 68% enrolled in the CGN. Trust was strongly related to enrollment. Less education, less satisfaction with cancer care, and individualistic rather than collective orientation were associated with lower trust. Education was also bivariately associated with enrollment, but mediation analysis indicated that the operational mechanism of education's influence on enrollment was through trust. PMID- 18417971 TI - Testing targeted approaches to enhance Cancer Genetics Network minority recruitment within Asian populations. AB - BACKGROUND/AIMS: Asian Americans have been underrepresented in cancer research. The purpose of this study was to evaluate the efficacy of a multiple arm recruitment approach in improving Asian recruitment into the Cancer Genetics Network (CGN). METHODS: 1,096 potential participants, identified through cancer registries located at University of California, Irvine (UCI) and Fred Hutchinson Cancer Research Center (FHCRC), were randomly assigned to receive one of four recruitment approaches. RESULTS: A 6.2% gain in Asian participation into the CGN was achieved over a 2-year period at FHCRC and UCI, which contributed a 2% CGN wide increase in overall Asian enrollment. Site-specific differences in recruitment success by study arm were observed. CONCLUSION: Novel recruitment approaches can assist in improving recruitment of Asian populations into cancer genetic research studies. PMID- 18417972 TI - Strategies and stakeholders: minority recruitment in cancer genetics research. AB - The Cancer Genetics Network (CGN) is one of a growing number of large-scale registries designed to facilitate investigation of genetic and environmental contributions to health and disease. Despite compelling scientific and social justice arguments that recommend diverse participation in biomedical research, members of ethnic minority groups continue to be chronically underrepresented in such projects. The CGN studies reported in this issue used strategies well documented to increase minority participation in research activities, including use of community-targeted materials, addressing community trust concerns, and the adoption of personalized and flexible research protocols. Here, we review the outcome of these efforts to increase minority recruitment to the CGN, and ask what lessons the findings suggest for future minority recruitment initiatives. PMID- 18417973 TI - Transition from cognitively impaired not demented to Alzheimer's disease: an analysis of changes in functional abilities in a dementia clinic cohort. AB - BACKGROUND: Patients with cognitive impairment no dementia (CIND) are at an increased risk of progression to Alzheimer's disease (AD). Whether subtle impairments in functional or social abilities at the CIND stage can predict progression to AD is not yet fully determined. We evaluated whether impairments on the Disability Assessment for Dementia (DAD) and Functional Rating Scale (FRS) can predict progression to AD. METHODS: We examined 70 patients with CIND from the ACCORD cohort having complete DAD and FRS baseline and 2-year follow-up data. MANCOVA adjusted for age, sex, education and baseline MMSE score compared the baseline and 2-year change in DAD and FRS scores in CIND patients who progressed to AD versus non-progressors. RESULTS: There were no significant differences between CIND progressors and non-progressors in baseline total DAD or FRS scores. FRS domain analysis revealed that greater impairment in social/occupational functioning significantly predicted progression, while there were no predictive DAD domains. In progressors, both DAD and FRS scores significantly declined over time with the largest changes in instrumental activities of daily living (IADL). CONCLUSION: While changes in IADL characterize the progression from CIND to AD, impairment in complex social-cognitive competency significantly predicts risk of progression and may mark early AD. PMID- 18417974 TI - Fetal cytomegalovirus infection associated with cerebral hemorrhage, hydrops fetalis, and echogenic bowel: case report. AB - We describe some fetal ultrasound findings associated with intrauterine cytomegalovirus (CMV) infection. We report a 38-year-old gravida 3, para 2 at 16 weeks of gestation who underwent ultrasound examination for anomaly screening. The scan revealed an extensive irregular echogenic area in the fetal brain, especially at the level of lateral ventricles, suggestive of intraventricular and cerebral hemorrhage. Cardiomegaly, hepatomegaly, and mild ascites as well as an echogenic bowel were demonstrated. Abnormal chromosomes and hemoglobin Bart disease were excluded by analysis of fetal blood. Follow-up ultrasound at 20 weeks of gestation showed frank hydrops fetalis, and termination of the pregnancy was performed based on the couple's decision, giving stillbirth to a male fetus weighing 450 g. Autopsy findings showed intracerebral hemorrhage (right cerebral hemisphere) and hydrops fetalis with hepatosplenomegaly. Microscopic investigation showed typical changes of CMV infection in several organs, including brain, thyroid gland, lung, liver, kidney, heart, pancreas, and placenta. Sonographically, the combination of hydrops fetalis, cerebral hemorrhage, and hyperechoic bowel should raise the possibility of a CMV infection, particularly in cases with no obvious cause of hydrops fetalis. PMID- 18417975 TI - What about informed consent in first-trimester ultrasound screening for Down syndrome? AB - OBJECTIVES: We evaluate the level of information and informed consent given for the screening of Down syndrome (DS) before the first-trimester ultrasound scan. We evaluate the nature of the medical information given and the patient's understanding. This makes it possible to analyze the existence of the necessary elements (level of information and consent) for the patients to exercise their autonomy as to their decision making during the first-trimester ultrasound scan. METHODS: A total of 350 patients were recruited prospectively in 2 hospitals. Patients were divided into 3 groups according to the results of maternal serum screening for DS. Group 1: Patients at high risk of having a child with DS with the second-trimester maternal serum screening (MSS) (risk >1/250). Group 2: Patients having a low risk (risk <1/250). Group 3: Patients who did not undergo maternal serum screening test. A questionnaire was completed before the medical consultation, to assess the quality of the medical consultation before the ultrasound, the level of prior knowledge and the maternal consent. RESULTS: We excluded 45 of the 350 questionnaires from the analysis because they were incomplete (13.1%). We therefore analyzed a total of 305 questionnaires. The three groups analysed were: Group 1: 89 patients (29.2%) identified as at high risk after MSS. Group 2: 137 patients (44.9%) identified as at low risk accordingly to MSS. Group 3: 79 women (25.9%) who did not undergo maternal serum screening test for diverse reasons. The frequency of genuine informed consent was significantly (p < 0.001) higher in group 3 (56.9%) than in group 1 (52.8%) or group 2 (32.1%). Finally, the number of completely uninformed patients was significantly higher in group 2 (18%) than in the other two groups (7.8% for group 1 and 6.3% for group 3, p < 0.001). With multiple regression analysis, we found that maternal consent depended on 4 independent components: (1) Educational level ( or =10). The negative responses to the various questions calls into question the legitimacy of consent for first-trimester ultrasound scans, particularly as concerns the measurement of nucal translucency. PMID- 18417976 TI - Risk of chromosome abnormalities in the presence of bilateral or unilateral choroid plexus cysts. AB - OBJECTIVES: To evaluate the rate of chromosome abnormalities in cases of uni- and bilateral choroid plexus cysts (CPCs). METHODS: A total of 10,875 ultrasound (US) examinations were performed in the second trimester, and 435 cases with CPC (4%) were found. After genetic counseling, 45 patients decided not to undergo karyotyping. The authors performed a chromosome analysis in 390 cases of CPCs. RESULTS: The total risk of chromosome abnormalities was 3.59% (n = 14) and risk of trisomies was 2.05% (n = 8). Trisomy 18 was found in 6 cases (1.54%), trisomy 21 in 1 case (0.26%), and trisomy 9 in 1 case (0.26%). The risk of 45,X karyotype was 0.77% (n = 3). One case of 47,XXY karyotype and 2 cases with other chromosome abnormalities were found. In 212 unilateral cases there were 7 with chromosome abnormalities (3.3%). In 178 bilateral cases there were 7 with abnormal karyotypes (3.93%). The CPC was associated with additional fetal US anomalies (with or without polyhydramnios/oligohydramnios) in 112 cases; chromosome abnormalities were detected in 4 cases (3.57%). 66 cases were associated with polyhydramnios/oligohydramnios but not with other fetal US anomalies; 3 cases of abnormal karyotypes were found (4.55%). The CPC was isolated in 212 cases and 7 cases were associated with chromosome disorders (3.3%). CONCLUSIONS: US plays an important role in prenatal diagnostics. Further genetic counseling is recommended in cases with CPCs. PMID- 18417977 TI - Feticide does not modify duration of labor induction in cases of medical termination of pregnancy. AB - OBJECTIVES: To compare the general characteristics and clinical outcome of women who underwent medical termination of pregnancy preceded or not by feticide. METHODS: A total of 146 women of diverse gestational ages, who had received a sonographic or genetic diagnosis of fetal abnormality incompatible with postnatal life and chose to legally terminate their pregnancy, had labor induced following feticide (n = 82) or without having feticide performed (n = 64). RESULTS: The number of obstetrical procedures was higher (p = 0.001) in the feticide group. Time of labor induction and complications were similar in both groups. CONCLUSION: Feticide does not alter the clinical obstetrical course of women undergoing medical termination of pregnancy, and poses no risk to maternal health regardless of gestational age. PMID- 18417978 TI - Skewed X-chromosome inactivation in Greek women with idiopathic recurrent miscarriage. AB - OBJECTIVES: To investigate if skewed X-chromosome inactivation (XCI) is associated with unexplained recurrent miscarriage (RM) in Greek women. METHODS: This was a prospective case-control study. A methylation-sensitive assay was used to investigate the X-inactivation pattern of women with unexplained RM and controls. RESULTS: Fifty-six of the 74 patients (75.7%) and 55 of 80 controls (68.8%) were informative. Among the informative cases, 6/56 (10.7%) women showed extreme XCI (>90%) and among the informative controls, 2/55 (3.6%) showed extreme XCI. CONCLUSIONS: In the present study, women with unexplained RM showed a statistically nonsignificant increase in skewed XCI prevalence (10.7%) compared with control women (3.6%; p = 0.271). PMID- 18417979 TI - Analysis of factors influencing the ultrasonic fetal weight estimation. AB - OBJECTIVE: The aim of our study was the evaluation of sonographic fetal weight estimation taking into consideration 9 of the most important factors of influence on the precision of the estimation. METHODS: We analyzed 820 singleton pregnancies from 22 to 42 weeks of gestational age. We evaluated 9 different factors that potentially influence the precision of sonographic weight estimation (time interval between estimation and delivery, experts vs. less experienced investigator, fetal gender, gestational age, fetal weight, maternal BMI, amniotic fluid index, presentation of the fetus, location of the placenta). Finally, we compared the results of the fetal weight estimation of the fetuses with poor scanning conditions to those presenting good scanning conditions. RESULTS: Of the 9 evaluated factors that may influence accuracy of fetal weight estimation, only a short interval between sonographic weight estimation and delivery (0-7 vs. 8-14 days) had a statistically significant impact. CONCLUSION: Of all known factors of influence, only a time interval of more than 7 days between estimation and delivery had a negative impact on the estimation. PMID- 18417980 TI - Human placental growth hormone is increased in maternal serum in pregnancies affected by Down syndrome. AB - OBJECTIVE: To evaluate the relationship between maternal serum levels of human placental growth hormone (hPGH) and fetal Down syndrome at gestational midtrimester. METHODS: We retrospectively analyzed samples of serum from 21 women with Down syndrome pregnancies detected at gestational midtrimester. The samples were obtained at 16-23 weeks' gestation during amniocentesis for fetal karyotyping. Sixty-two serum samples were used as controls, which were obtained at 16-23 weeks' gestation from women with singleton, uncomplicated pregnancies, who gave birth to healthy neonates with a birth weight appropriate for gestational age. The hPGH levels were measured by a solid-phase immunoradiometric assay using 2 different epitopes. RESULTS: The median hPGH values in the serum of the Down-syndrome-affected pregnancies were significantly higher (p < 0.05) than those of the normal pregnancies at 16-23 weeks' gestation: the median value in the serum was 9.4 ng/ml (5th to 95th percentiles = 1.49-39.03) versus 4.7 ng/ml (0.53-7.88). CONCLUSION: The hPGH levels in maternal serum were found to be higher at 16-23 weeks' gestation in pregnancies affected by fetal Down syndrome. Further investigation is needed to examine if maternal serum hPGH could be used as an additional marker in prenatal screening of Down syndrome at gestational midtrimester. PMID- 18417981 TI - Fetal loss after amniocentesis in a series of 5,780 procedures. AB - OBJECTIVES: Counseling on prenatal diagnosis requires accurate knowledge of the associated risks, including fetal loss. The objective of our study was to assess this risk of amniocentesis in a single center with several operators. METHODS: This retrospective analysis concerns only women with singleton pregnancies who underwent amniocentesis between 14(+0) and 23(+6) weeks' gestation. RESULTS: During this 4.5-year period, 5,780 amniocenteses were performed, of which we analyzed 5,319. The rate of fetal loss was 70 in 4,858 tests (1.4%), with a lost to-follow-up rate of 3.8%. CONCLUSION: Our results for fetal loss are comparable to those in the largest series with fewer operators already published. PMID- 18417982 TI - Vaginal delivery using cranioclasia following prenatal diagnosis of a giant fetal intracranial teratoma. AB - Intracranial teratoma, although a rare disease, is nonetheless the most common of the congenital tumors of the central nervous system in the neonatal period. It can be diagnosed antenatally by ultrasound. The prognosis for fetal forms of teratoma is very grim. We report here the case of an immature teratoma identified at 26 weeks' gestation and discuss its obstetric consequences, in particular, the technical procedures for a medically-indicated termination of pregnancy in view of the massive increase in head circumference associated with this tumor. PMID- 18417983 TI - Fetal manifestation of the Fine-Lubinsky syndrome. Brachycephaly, deafness, cataract, microstomia and mental retardation syndrome complicated by Pierre-Robin anomaly and polyhydramnios. AB - OBJECTIVES: We report on a female fetus of 24 weeks gestational age with Fine Lubinsky syndrome (FLS), representing the 7th case published so far. METHODS: Prenatal ultrasound was performed at 22+1 weeks of gestation and thorough postmortem examination was made after termination of pregnancy. RESULTS: The diagnosis of FLS in the fetus was based on characteristic features that were already apparent in early prenatal life, such as growth deficiency, brachycephaly, flat face with associated dysmorphic signs, microstomia and cataract, while deafness and mental retardation, which are syndrome-specific functional disorders and evident only postnatally, could not be taken into account. CONCLUSIONS: This case demonstrates the diagnostic problems in fetal syndromology if syndrome-specific features are not yet recognizable and additional complications occur that had not been observed in this disorder. PMID- 18417984 TI - Adverse outcome of pregnancies with extremely high levels of maternal serum human chorionic gonadotropin. AB - BACKGROUND: The associated risk of elevated levels of maternal serum human chorionic gonadotropin (MShCG) with pregnancy complications was reported in many studies. However, the outcome of pregnancies with extremely high levels of MShCG was never independently studied. METHODS: We report on 6 out of 45,990 studied patients with extremely high levels of MShCG (>15 multiples of the medians) analyzed during the second trimester. RESULTS: Although our patient population was composed of more Jewish than Arab pregnant women, all those patients were Arabs. Overall, the prognosis of those pregnancies was poor. In 1 case, an antepartum fetal death occurred, 2 had premature deliveries (one of the newborns had severe failure to thrive), and 2 delivered small for gestational age babies. In 5 of these 6 cases, no specific diagnosis was established. One case was complete hydatidiform mole with a coexisting normal fetus. CONCLUSIONS: We recommend that these patients undergo counseling in which the predicted outcome will be described. In addition, a follow-up of high-risk pregnancy should be implemented: sonographic evaluation should be performed, initially to rule out a molar gestation. The patients should then be followed for growth restriction and they should be monitored to rule out other pregnancy complications such as premature labor and antepartum fetal death. Finally, the overrepresentation of Arabs among our affected patients raises the question of a possible genetic tendency for increased MShCG levels especially in the extreme level group. PMID- 18417985 TI - Fetal cardiocentesis: a review of indications, risks, applications and technique. AB - OBJECTIVES: To review the indications, applications and technique of fetal cardiocentesis. METHODS: Review of published case reports and case series of fetal cardiocentesis utilizing the PubMed search engine of the National Library of Medicine. RESULTS: Case reports and case series demonstrate that fetal cardiocentesis may be an alternative method by which to facilitate prenatal diagnosis, intravascular therapy, multifetal and selective fetal reduction and in utero therapy of congenital heart disease. However, procedure-associated risk is higher than with cordocentesis and may limit use of this procedure. CONCLUSIONS: Fetal cardiocentesis may be a reasonable option to obtain fetal intravascular access and facilitate therapeutic interventions when cordocentesis fails or is not feasible. However, expected benefit must clearly outweigh the procedure associated risk. PMID- 18417986 TI - Organic acid concentrations in amniotic fluid found in normal and Down syndrome pregnancies. AB - INTRODUCTION: Organic acids were examined from normal and Down syndrome pregnancies to identify possible differences between the amniotic fluid from fetuses with Down Syndrome compared with that of normal fetuses. MATERIALS AND METHODS: Amniotic fluids were obtained from prior amniocenteses. Forty-one normal and 22 Down syndrome specimens were assayed using gas chromatography/mass spectrometry. RESULTS AND DISCUSSION: 5-hydroxycaproate, methylsuccinate, alpha ketoglutarate, and adipate were significantly elevated in Down syndrome, suggesting riboflavin deficiency. Phenylpyruvate was also significantly elevated in fetuses with Down syndrome. Phenylpyruvate inhibits the metabolism of tetrahydrobiopterin, which is necessary for neurotransmitter metabolism. Elevated phenylpyruvate is consistent with previous research, suggesting a disturbance of tetrahydrobiopterin metabolism in Down syndrome. CONCLUSION: Organic acid markers for B2 deficiency are elevated in the amniotic fluid of fetuses with Down syndrome. Elevation of phenylpyruvate may impair neurotransmitter metabolism. Organic acid markers for B12 levels are not different between the Down syndrome and normal group. PMID- 18417987 TI - Aneurysm. Commentary on Grethel EJ et al: Prenatal and Postnatal Management of a Patient with Pentalogy of Cantrell and Left Ventricular Aneurysm (Fetal Diagn Ther 2007;22:269-273). PMID- 18417988 TI - Does congenital diaphragmatic hernia associated with bronchopulmonary sequestration portend a better prognosis? AB - INTRODUCTION: Congenital diaphragmatic hernia (CDH) continues to be a devastating disease in the newborn population, with well-documented morbidity and mortality. Bronchopulmonary sequestration is a separate congenital defect that has been associated with CDH. While the association of sequestration with CDH has been reported to be as high as 30-40%, the prognosis associated with the two simultaneous defects is unknown. We reviewed our experience to evaluate if prognosis was better in the CDH infants with associated bronchopulmonary sequestration. METHODS: Institutional approval was obtained. Our institutional database was examined from August 1995 to August 2005, identifying all mothers carrying fetuses with pulmonary masses and/or CDH and all neonates treated with bronchopulmonary sequestration and/or CDH. Patients who had both CDH and sequestration were identified by prenatal ultrasound reports, postnatal radiographs, and operative and pathology reports. RESULTS: 16 patients were identified in the fetal or neonatal period with concomitant diagnoses of CDH and bronchopulmonary sequestration. Of those proceeding to delivery, 6 expired and 6 survived. The presence of liver herniation and low lung-to-head ratio on antenatal ultrasound correlated with mortality. However, 2 patients survived with very low lung-to-head ratio that would usually be associated with 100% mortality at our institution. Two diagnoses of bronchopulmonary sequestration were reversed after final pathology revealed liver tissue. CONCLUSION: Given the limited series, we cannot conclude that bronchopulmonary sequestration confers an anatomic advantage to patients that have CDH. We did observe survivors in this group that, given their antenatal predictors of CDH severity, would ordinarily have dismal prognosis. The presence of a sequestration may be protective in a subset of patients with severe CDH, or may confound our antenatal predictors of disease severity in these patients. PMID- 18417989 TI - Pyridoxine-related metabolite concentrations in normal and Down syndrome amniotic fluid. AB - INTRODUCTION: Some studies of children with Down syndrome have found mild abnormalities in the metabolism of pyridoxine (vitamin B(6)); therefore the present question is whether such abnormalities might also be present in the amniotic fluid of fetuses with Down syndrome. MATERIALS AND METHODS: Archived specimens of amniotic fluid were obtained from chromosomally normal and from fetuses with Down syndrome. Gas chromatography/mass spectrometry quantitized B related metabolites, including oxalate, xanthurenate, kynurenine and 4-pyridoxic acid. RESULTS: Oxalate, a marker of pyridoxine deficiency, was elevated in the amniotic fluid of fetuses with Down syndrome. This result was statistically significant. The other marker results were not statistically significant. CONCLUSION: A marker of pyridoxine deficiency, oxalate is elevated in the amniotic fluid of fetuses with Down syndrome. These results in amniotic fluid are consistent with previous studies done in the urine of young children. PMID- 18417990 TI - Liver position is a prenatal predictive factor of prosthetic repair in congenital diaphragmatic hernia. AB - OBJECTIVE: To determine whether any common maternal-fetal variable has prenatal predictive value of prosthetic repair in congenital diaphragmatic hernia. METHODS: This was a 5-year single-center retrospective review of fetal congenital diaphragmatic hernia referrals. Multiple prenatal variables were correlated with the need for a prosthetic repair. Statistical analyses were by Fisher's exact and Mann-Whitney U-tests, as appropriate (p < 0.05). RESULTS: Fetal liver position was a predictor of prosthetic repair. The presence or absence of liver herniation was correlated with prosthetic repair rates of 83.3 and 23.1%, respectively (p < 0.001). All patients with moderate/severe liver herniation required a prosthetic patch. CONCLUSION: Liver herniation has prenatal predictive value for the need for prosthetic repair in congenital diaphragmatic hernia. This finding should be valuable during prenatal counseling for clinical trials of engineered diaphragmatic repair. PMID- 18417991 TI - Under-expression of CD24 in pre-eclamptic placental tissues determined by quantitative real-time RT-PCR. AB - BACKGROUND: Pre-eclampsia is a pregnancy-related disorder present in about 5-7% of all pregnancies. CD24 expression was recently reported in different diseases, while it has not yet been determined in pre-eclamptic placental tissues. METHODS: We collected placental tissues from pre-eclamptic (n = 16) and healthy pregnancies (n = 16). We used the quantitative real-time PCR method with a primer probe system for determination of CD24 gene expression. RESULTS: We measured CD24 concentrations of 18.94 +/- 26.86 ng/microl in the pre-eclamptic and 53.85 +/- 92.05 ng/microl in the healthy placental tissues (p = 0.03). CONCLUSIONS: The quantitative real-time PCR method is suitable to determine CD24 expression in placental tissues. We suppose the low expression of CD24 may cause the enhanced immune reaction and could play a role in the abnormal development of placenta in pre-eclampsia. PMID- 18417992 TI - A prenatal diagnosed case of primitive neuroectodermal tumor. AB - Congenital brain tumors presenting in the first year of life account for 1.4-8.5% of all childhood brain tumors. Only 18% of congenital brain tumors are diagnosed before or at delivery. Primitive neuroectodermal tumor (PNET) is a highly malignant, small, blue-cell tumor which is characterized by early recurrence, metastasis, and high mortality. It makes up 13% of all fetal and neonatal brain tumors. Prenatal diagnosis of PNET or other congenital brain tumors is important because the presence of tumors may alter the time and mode of delivery and postnatal care planning. A prenatal case of PNET is described that was diagnosed with ultrasonography and magnetic resonance imaging; the case was confirmed histopathologically at postmortem examination. PMID- 18417993 TI - Fetal spina bifida repair--current trends and prospects of intrauterine neurosurgery. AB - Myelomeningocele is a common dysraphic defect leading to severe impairment throughout the patient's lifetime. Although surgical closure of this anomaly is usually performed in the early postnatal period, an estimated 330 cases of intrauterine repair have been performed in a few specialized centers worldwide. It was hoped prenatal intervention would improve the prognosis of affected patients, and preliminary findings suggest a reduced incidence of shunt-dependent hydrocephalus, as well as an improvement in hindbrain herniation. However, the expectations for improved neurological outcome have not been fulfilled and not all patients benefit from fetal surgery in the same way. Therefore, a multicenter randomized controlled trial was initiated in the USA to compare intrauterine with conventional postnatal care, in order to establish the procedure-related benefits and risks. The primary study endpoints include the need for shunt at 1 year of age, and fetal and infant mortality. No data from the trial will be published before the final analysis has been completed in 2008, and until then, the number of centers offering intrauterine MMC repair in the USA is limited to 3 in order to prevent the uncontrolled proliferation of new centers offering this procedure. In future, refined, risk-reduced surgical techniques and new treatment options for preterm labor and preterm rupture of the membranes are likely to reduce associated maternal and fetal risks and improve outcome, but further research will be needed. PMID- 18417995 TI - Gasless fetoscopy: a new approach to endoscopic closure of a lumbar skin defect in fetal sheep. AB - OBJECTIVE: To develop a new endoscopic approach to the correction of a myelomeningocele-like defect in fetal sheep. METHODS: The fetuses of 9 pregnant ewes, with an average gestational age of 115 days, were subjected to a 3.0 x 2.0 cm removal of the skin over the lumbar spine, performed through hysterotomy. The uterus was closed, and three 5-mm endoscopic cannulas, without valve mechanisms, were inserted. In the pilot phase (2 animals), we initially worked exclusively in the amniotic fluid space. In the study phase, we partially withdrew the fetus from the amniotic fluid to completely expose its back. By simply allowing air to enter the amniotic cavity (without gas injection), a working space was created using a uterine lift device. The skin around the defect was dissected, and a biosynthetic cellulose material was applied to cover the area. A continuous suture of the skin was performed to completely hide the material. RESULTS: The combined air/fluid space allowed the skin to be successfully closed in 6 out of 7 cases in the study phase. All fetuses were alive at the end of the procedures. Time to complete the endoscopic part of the procedure fell from 3 to 1 h by the end of this series. Premature birth occurred in 2 of the 4 cases allowed to continue with the pregnancy. CONCLUSION: A new gasless fetoscopic surgery technique was developed as an alternative to current techniques used for fetal endoscopic surgery. PMID- 18417994 TI - Fetal urogenital sinus with consecutive hydrometrocolpos because of labial fusion: prenatal diagnostic difficulties and postpartal therapeutic management. AB - OBJECTIVE: To elucidate the differential diagnoses of tumorous dilations in the fetal pelvic region detected by prenatal ultrasound and the postnatal management demonstrated on a fetus with 29 weeks of gestation with a retrovesical located bottle-like cystic structure measuring 54 x 31 x 27 mm within the pelvis. Postnatal findings were a labial fusion and a consecutive hydrometrocolpos due to a urethrovaginal fistula. METHOD: Case report of a fetus affected by an intricate cloacal anomaly. RESULTS: The long-term prognosis for this nonsyndromic form of hydrometrocolpos without any other structural defects or organic failures after operative sanitation is excellent. Final reconstruction is planned at about 12 months of age. CONCLUSION: Prenatal diagnosis of tumorous dilations in the fetal pelvic region often involves difficulties because of numerous differential diagnoses and possible presentation in late pregnancy. Magnetic resonance imaging could be a useful complementary tool for assessing these anomalies when ultrasonography is inconclusive. In some cases, the final diagnosis cannot be confirmed until after delivery. PMID- 18417996 TI - Aberrant venous flow measurement may predict the clinical behavior of a fetal extralobar pulmonary sequestration. AB - OBJECTIVE: Fetal extralobar pulmonary sequestration (EPS) is sometimes complicated by a massive pleural effusion, leading to tension hydrothorax and fetal hydrops. The goal of this study was to examine sonographic signs of venous obstruction in fetal EPS with or without pleural effusion. METHODS: Records of fetal ultrasound from 6 patients with EPS were reviewed with special attention to aberrant arterial and venous flow. The results were correlated with their clinical outcomes. RESULTS: Four of the 6 cases (cases 1-4) were complicated by massive pleural effusion and required fetal thoracentesis; thoracoamniotic shunt placement was required in 3 of these 4 patients (cases 1-3). The other 2 patients (cases 5 and 6) were not associated with pleural effusion despite the comparable size of the mass and did not require any treatment, either prenatally or postnatally. In cases 1-3, aberrant venous flow was difficult to detect and, even when detected, the arterial-to-venous flow velocity ratio was >6. This is in contrast to the uncomplicated cases 5 and 6 in whom aberrant venous flow was easily detected with an arterial-to-venous flow velocity ratio of 2-3. Arterial to-venous flow velocity ratios of 3-6 were observed in case 4. This case was complicated by pleural effusion but not by fetal hydrops. CONCLUSIONS: These data support the hypothesis that venous obstruction is related to the production of pleural effusion in fetal EPS. Ample flow in the aberrant vein may indicate benign clinical behavior, while difficulty in detecting aberrant venous flow may be correlated with the development of massive pleural effusion. PMID- 18417997 TI - Evaluation of normal fetal branch pulmonary artery diameters measured by ultrasonography: a comparison with congenital diaphragmatic hernia. AB - OBJECTIVE: The aim of this study was to examine the size of fetal branch pulmonary artery (PA) diameters in normal growth fetuses. METHOD: Fetal PA diameters were measured in 175 normal fetuses between 18 and 40 weeks of gestation from 2005 to 2006. In addition, 4 fetuses with left-sided congenital diaphragmatic hernia (CDH) from 2001 to 2006 were retrospectively reviewed. Branch PA diameters were measured from a cross-sectional image at the level of the three-vessel view (main PA, ascending aorta and superior vena cava) to demonstrate the long axis of both branch PAs whenever possible. RESULTS: Both the left and right PA diameters were found to correlate strongly with the advancing gestational age (r = 0.78, p < 0.01, respectively). Left pulmonary artery to main pulmonary artery (LPA/MPA) ratio and right pulmonary artery to main pulmonary artery (RPA/MPA) ratio were calculated. These two parameters were almost constant throughout gestation. LPA in fetal left CDH was smaller than control. Both LPA/MPA and RPA/MPA were within the normal range in most cases. CONCLUSIONS: It is important to establish the normal range of branch PA diameters and it appears to be useful to compare the branch PA diameters in normal fetuses with that in cases of pulmonary hypoplasia. PMID- 18417998 TI - Screening advances and diagnostic choice: the problem of residual risk. AB - OBJECTIVE: Over the past decade some authorities have suggested that advanced screening methodologies obviate the need for more invasive, diagnostic procedures. Data on Down syndrome (DS) births for Colorado from 1989 to 2005 were used to examine the implications of a decreasing use of amniocentesis. METHODS: Publicly available, State of Colorado Department of Public Health data on DS birth rates for women were compared to amniocentesis use at Colorado's largest prenatal diagnostic center. Longitudinal changes on DS birth rates by maternal age (>35 and <35), and utilization of amniocentesis. RESULTS: In Colorado, from 1989 to 2005, the rate for DS births for women 35+ rose considerably, while <35, rates remained stable (Cochran-Armitage test, p < 0.001). An autocorrelation corrected test yielded a significant negative relationship between amniocentesis use (in 1,000 s) and AMA DS rates (b = -11.30; p < 0.006; DW = 1.55). Confounding explanations involving sampling problems, socio-demographic factors, political conservatism and prevention orientation do not appear to account for these results. CONCLUSIONS: Replacement of definitive diagnosis with screening tests must be implemented with caution, particularly when using technologies with wide individual operator-dependent variability. Screening paradigms when performed with accuracy can markedly improve assessment of risks, but caution must be used in presenting negative screening results to women who still have a relatively high residual risk after a negative screen, and more generally in the displacement of technologies that provide definitive answers. PMID- 18418000 TI - TNFalpha -308 C-->t and -863 C-->a polymorphisms and spermiogram characteristics. AB - BACKGROUND: Genetic factors may play a role in male infertility. METHODS: In a prospective case-control study, we assessed the allele and genotype frequencies of the TNFalpha -308 C-->T and -863 C-->A polymorphisms, detected by PCR of sperm DNA, of 577 Caucasian men recruited in an infertility clinic. Semen sampling was performed and spermiogram results were correlated to genetic data. RESULTS: The allele frequencies of the TNFalpha -308 C-->T and -863 C-->A polymorphisms were not significantly different between non-normozoospermic (n = 447) and normozoospermic (n = 130) men [758/894 (85%) and 134/894 (15%) vs. 213/269 (82%) and 43/260 (18%), p = 0.5, odds ratio (OR) 1.1, 95% confidence interval (CI) 0.74 1.76, and 749/894 (84%) and 145/894 (16%) vs. 212/260 (82%) and 48/260 (18%), p = 0.4, OR 1.2, 95% CI 0.78-1.76, respectively]. The genotype frequencies of the TNFalpha -308 C-->T and -863 C-->A polymorphisms were also not significantly different between non-normozoospermic and normozoospermic men. In addition, mutant alleles were not overrepresented in subgroups of men with the oligoasthenoteratozoospermia syndrome and asthenozoospermia. CONCLUSION: The TNFalpha -308 C-->T and -863 C-->A polymorphisms are not associated with spermiogram characteristics and do not represent molecular markers for genetic susceptibility to male infertility. PMID- 18418001 TI - Strategies for conditional two-locus nonparametric linkage analysis. AB - In this article we deal with two-locus nonparametric linkage (NPL) analysis, mainly in the context of conditional analysis. This means that one incorporates single-locus analysis information through conditioning when performing a two locus analysis. Here we describe different strategies for using this approach. Cox et al. [Nat Genet 1999;21:213-215] implemented this as follows: (i) Calculate the one-locus NPL process over the included genome region(s). (ii) Weight the individual pedigree NPL scores using a weighting function depending on the NPL scores for the corresponding pedigrees at speci fi c conditioning loci. We generalize this by conditioning with respect to the inheritance vector rather than the NPL score and by separating between the case of known (prede fi ned) and unknown (estimated) conditioning loci. In the latter case we choose conditioning locus, or loci, according to prede fi ned criteria. The most general approach results in a random number of selected loci, depending on the results from the previous one-locus analysis. Major topics in this article include discussions on optimal score functions with respect to the noncentrality parameter (NCP), and how to calculate adequate p values and perform power calculations. We also discuss issues related to multiple tests which arise from the two-step procedure with several conditioning loci as well as from the genome-wide tests. PMID- 18418002 TI - Tracking adult neovascularization during ischemia and inflammation using Vegfr2 LacZ reporter mice. AB - The vascular endothelial growth factor/vascular endothelial growth factor receptor 2 (VEGF/VEGFR-2) signal transduction system plays a key role during embryonic vascular development and adult neovascularization. In contrast to many endothelial genes, VEGFR-2 is expressed at low levels in most adult vessels but is strongly upregulated during neovascularization, leading to a pro-angiogenic response. Here, we analyzed the activity of regulatory sequences of the murine Vegfr2 gene during neovessel formation in vivo under ischemic and inflammatory conditions. Hindlimb ischemia was induced in transgenic mice, expressing the LacZ reporter gene under the control of Vegfr2 promoter/enhancer elements. Most vessels in the ischemic muscle tissue showed strong endothelium-specific reporter gene expression, whereas nearly no LacZ-expressing capillaries were observed in untreated control tissue. Cutaneous punch wounds were created to induce angiogenesis under inflammatory conditions, leading to robust LacZ expression in the majority of the blood vessels in the wound tissue. Since the cornea is physiologically avascular, the functionality of these promoter/enhancer elements exclusively in newly formed vessels was confirmed using the cornea micropocket assay. Taken together, our results show that these Vegfr2 regulatory elements are active during adult neovessel formation in general. Therefore, these sequences may prove to be valuable targets for novel endothelium-specific anti-angiogenic as well as pro-angiogenic treatment strategies. They may especially allow directing therapeutic gene expression to sites of adult neovascularization. Moreover, the Vegfr2/LacZ reporter mice represent a powerful model to generally analyze the transcriptional control mechanisms involved in the induction of Vegfr2 expression during adult neovascularization. PMID- 18418003 TI - Gene expression profiles of vascular smooth muscle show differential expression of mitogen-activated protein kinase pathways during captopril therapy of heart failure. AB - Congestive heart failure (CHF) is characterized by increased vascular tone and an impairment in nitric-oxide-mediated vasodilatation. We have demonstrated that the blunted response to nitric oxide is due, in part, to a reduction in the leucine zipper-positive isoform of the myosin-targeting subunit (MYPT1) of myosin light chain phosphatase. Additionally, we have shown that angiotensin-converting enzyme inhibition, but not afterload reduction with prazosin, preserves leucine-zipper positive MYPT1 isoform expression in vascular smooth muscle cells and normalizes the sensitivity to cGMP-mediated vasodilatation. We therefore hypothesized that in CHF, growth regulators and cytokines downstream of the angiotensin II receptor are involved in modulating gene expression in vascular tissue. Rats were divided into control and captopril-treated groups following left coronary artery ligation. Gene expression profiles in the aorta and portal vein at baseline and 2 and 4 weeks after myocardial infarction (MI) were analyzed using microarray technology and quantitative real-time PCR. After MI, microarray analysis revealed differential mRNA expression of 21 genes in the aorta of captopril-treated rats 2 and 4 weeks after surgery when compared to gene expression profiles at baseline and without captopril therapy. Real-time PCR demonstrated that captopril suppressed the expression of protein kinases in the angiotensin-II-mediated mitogen-activated protein kinase signaling pathway, including Taok1 and Raf1. These data suggest that in CHF, captopril therapy modulates gene expression in vascular smooth muscle, and some of the beneficial effects of ACE inhibition may be due to differential gene expression in the vasculature. PMID- 18418004 TI - Prevalence and risk of hypertension in renal disease--data from the Czech Registry of Renal Biopsies. AB - BACKGROUND/AIMS: We intended to compare the risk and prevalence of hypertension in patients undergoing renal biopsy with those of the general population and to investigate the possible effects of various independent factors (age, sex and degree of renal insufficiency) on the prevalence of hypertension. METHODS: Data obtained within the Czech Registry of Renal Biopsies over an 8-year period (1995 2002) were statistically evaluated and compared with those of the general population obtained within the Post-MONICA Study conducted in 2000/2001. RESULTS: Hypertension was present in 1,839 out of a total of 3,601 renal patients (51.1%). The risk of hypertension in the patients with renal disease was increased in all age groups compared with the general population (OR = 1.3-5.3). The prevalence of hypertension increased significantly with age, serum creatinine and proteinuria (p < 0.001). Male sex was identified as an independent risk factor for the presence of hypertension (p < 0.01). Sex, age and glomerular filtration rate were shown to be more important determinants of the risk of hypertension than the underlying biopsy-proven diagnosis itself. CONCLUSION: Hypertension occurs as a common complication of renal disease, even in its early stages, and irrespective of the histological finding. PMID- 18418005 TI - Efficacy and (pharmaco)kinetics of one single dose of rasburicase in patients with chronic kidney disease. AB - AIMS: Hyperuricemia is a risk factor associated with cardiovascular and renal disease. Recently, rasburicase, a recombinant urate oxidase, has been developed for the treatment of hyperuricemia in patients with primarily hematological malignancies. We studied the pharmacokinetics and metabolism of rasburicase in the treatment of chronic asymptomatic hyperuricemia in chronic kidney disease (CKD) patients. MATERIALS AND METHODS: We studied 9 CKD patients with hyperuricemia, whose mean serum acid concentration was 10.2 (range 8.3-15.8) mg/dl. No study subject was taking allopurinol (3/9 are allopurinol intolerant). Patients were treated with rasburicase (0.2 mg/kg/day) in single dose by intravenous infusion over a 30-min period. Serum samples were collected after 1, 4, 8, 24, 48 and 72 h, after 1 week, and after 1 month. To evaluate the efficacy of rasburicase, plasma and urinary concentrations of uric acid were determined by the standard method; the plasma activity of rasburicase was determined using a new assay developed by our laboratory (chromatography-mass method, a colorimetric 96-well microtiter plate assay). RESULTS: All the treated patients experienced a rapid reduction in their plasma uric acid concentration. Data showed an undetectable value within 1 h of treatment. The rasburicase effect ended after 50 h, with a slow increase in the plasma level of uric acid. CONCLUSION: A single dose of rasburicase is highly effective and well tolerated in the treatment of hyperuricemia in selected CKD patients. PMID- 18418006 TI - Depressive symptoms and proinflammatory cytokine levels in chronic renal failure patients. AB - BACKGROUND/AIMS: Cytokine secretion is known to play an important role in the pathophysiology of depression, and levels of proinflammatory cytokines are increased in chronic renal failure (CRF) patients. The objective of this study was to examine the correlation between levels of proinflammatory cytokines in CRF patients and degree of depression. METHODS: 31 patients on hemodialysis, 31 patients on continuous ambulatory peritoneal dialysis, and 31 conservatively managed chronic kidney disease (CKD) patients were enrolled in this study. Depressive symptoms were measured with the Beck Depression Inventory (BDI), and 'elevated symptoms of depression' were defined as a BDI score > or =17. IL-6 and TNFalpha cytokine levels were measured by ELISA. RESULTS: 'Elevated symptoms of depression' occurred in 37 of 93 patients (40%). IL-6 and TNFalpha levels were not significantly different among CRF patients with and without elevated depressive symptoms (p = 0.937 and p = 0.414, respectively). When analyzed by treatment subgroup, proinflammatory cytokine levels were not significantly different in patients with and without elevated symptoms of depression. CONCLUSION: In patients with CRF, elevated symptoms of depression were not associated with increased cytokine levels. PMID- 18418007 TI - Effect of vagal nerve stimulation on systemic inflammation and cardiac autonomic function in patients with refractory epilepsy. AB - OBJECTIVE: Recent data suggest that vagus nerve stimulation (VNS) can inhibit cytokine release by inflammatory cells. Accordingly, an association between impaired cardiac parasympathetic function, as assessed by heart rate variability (HRV), and increased markers of inflammation has recently been reported. In this study we assessed the effect of direct left VNS on inflammatory markers and HRV in patients with refractory epilepsy. METHODS: A 24-hour electrocardiogram Holter recording was performed both at baseline and after 3 months of left VNS in 8 patients (age 32 +/- 24 years, 2 men) who underwent implantation of a VNS device because of refractory epilepsy. Tumor necrosis factor-alpha, interleukin-6 and C reactive protein serum levels were measured, as markers of inflammation, at the same times. RESULTS: No significant changes were found after 3 months of left VNS, compared to baseline, both for HRV variables and inflammatory markers. Also, no consistent correlation could be demonstrated between HRV parameters and inflammatory markers in these patients. CONCLUSIONS: Our data in epileptic patients without cardiovascular disease failed to show a significant effect of left VNS on cardiac autonomic function and on systemic inflammation at short-term follow-up. PMID- 18418008 TI - Pulmonary carcinoid tumours: indolent but not benign. AB - BACKGROUND: The aim of this retrospective study was to analyse the malignant behaviour of low-grade pulmonary neuroendocrine tumours (NETs) treated at our institution. PATIENTS AND METHODS: We reviewed 48 consecutive patients with pulmonary NETs referred to our Medical Oncology Unit between 1998 and 2006, including 33 subjects with typical carcinoids (TCs) and 15 with atypical carcinoids (ACs). RESULTS: At diagnosis, there were 37 metastatic and 11 non metastatic patients. Medical treatments used were somatostatin analogues, combined chemotherapy, within study protocols, 5 fluorouracil/dacarbazine/epiadriamycin (FDE), and oxaliplatin plus capecitabine (XELOX). Median disease-free survival was 72 months for the TC patients and 38 months for the AC patients. Actuarial 5-year survival was 93% for those with TCs and 73% for those with ACs. The mean overall survival was 68 months for the non metastatic patients (78 months for TC patients and 58 months for AC patients) and 36 months for patients with advanced disease (42 and 32 months, respectively). CONCLUSION: Cell type is the strongest determinant of prognosis, and the degree of malignancy increases from TCs to ACs. Moreover, the prognosis of metastatic pulmonary carcinoids is not as good as expected. Our analysis suggests that patients with advanced disease should receive first-line therapy with a somatostatin analogue, with chemotherapy regimens (FDE, XELOX) used in progressing cases. PMID- 18418009 TI - Preoperative treatment combining capecitabine with radiation therapy in rectal cancer: a GERCOR Phase II Study. AB - OBJECTIVE(S): To assess efficacy and tolerability of preoperative capecitabine chemoradiation in rectal cancer. METHODS: Patients received radiotherapy 45 Gy in 25 fractions over 5 weeks and capecitabine 825 mg/m(2) twice daily throughout radiotherapy. Surgery was performed 5-7 weeks after radiotherapy. The primary endpoint was pathological complete response, secondary endpoints were downstaging and tolerability. RESULTS: Fifty-one patients were enrolled in a phase II study, median age 62 years (range 35-78). Sixty-three percent of tumours involved the lower third of the rectum, 45% were fixed. The median delivered radiotherapy dose was 44.8 Gy (range 39.6-45.0 Gy) over 33-49 days. The treatment-related grade 3 adverse events were diarrhoea (12%), skin reactions (8%) and asthenia (8%), with no grade 4 toxicity. Fifty patients underwent surgery (29 conservative) and 1 patient refused. The pathological complete response rate was 20% and a further 10% of patients had minimal residual disease. Additional tumour downstaging was seen in 28% of patients and the sphincter preservation rate was 58%. CONCLUSIONS: Preoperative capecitabine chemoradiation is well tolerated and its efficacy supports further exploration, both as a single agent and as part of new therapeutic strategies. PMID- 18418010 TI - Haematological evaluation of weekly therapy with topotecan for the treatment of recurrent ovarian cancer resistant to platinum-based therapy. AB - Topotecan is indicated in the treatment of advanced-stage ovarian cancers refractory to prior platinum-based regimen. The aim of this study was to compare the standard therapeutic strategy with a novel strategy of weekly administration of topotecan. The primary endpoints were dose density and overall tolerance. This retrospective cohort study included patients with ovarian cancer in relapse. During a first period (1998-2001), 24 patients received the standard topotecan dose of 1.5 mg/m(2)/day for 5 consecutive days with a 3-week interval between each treatment course. During a second period (2003-2006), 21 patients received a weekly topotecan dose of 4 mg/m(2) for 3 weeks out of every 4. Grades III and IV haematological toxicities were more frequent with the standard strategy (p < 0.05), even after adjustment of the prescription of erythropoietin and G-CSF. With the weekly strategy, an increase in dose density and a reduction in the number of delayed doses were observed. No significant difference between the 2 strategies was found in terms of response to the treatment and specific survival. This study suggests that the weekly administration of topotecan 4 mg/m(2), for 3 weeks out of every 4, results in a better maintenance of dose density and a reduction in haematological toxicity. PMID- 18418011 TI - Assessment of baseline clinical predictive factors of response to cetuximab irinotecan in patients with irinotecan-refractory metastatic colorectal cancer. AB - OBJECTIVE: To identify easily available predictive factors of response to cetuximab-irinotecan in patients with irinotecan-refractory metastatic colorectal cancer. METHODS: Retrospective analysis of patients treated with cetuximab (400 mg/m(2) in week 1, 250 mg/m(2) in subsequent weeks) plus irinotecan (180 mg/m(2) every 2 weeks). We assessed demographic data, prior response to chemotherapy, number of metastatic sites, disease and metastatic disease durations, irinotecan free interval and tumoral immunohistochemical epidermal growth factor receptor status. RESULTS: We analyzed 311 patients. Objective response rate under cetuximab-irinotecan was 26%. In univariate analysis, prior response to irinotecan, presence of only 1 metastatic site, disease duration, metastatic disease duration and irinotecan-free interval equal or above median (24, 18 and 1.8 months, respectively) were predictive of response to cetuximab-irinotecan. Multivariate analysis confirmed independent predictive value of prior response to irinotecan, number of metastatic sites and disease duration. CONCLUSION: Prior response to irinotecan, number of metastatic sites and disease duration may contribute to better select patients suitable for cetuximab-irinotecan therapy. PMID- 18418012 TI - Clinical outcome of gastric cancer patients with bone marrow metastases. AB - BACKGROUND: Gastric cancer with bone marrow metastases is known to pursue a rapidly deteriorating clinical course. We conducted a retrospective analysis to evaluate clinical manifestations and prognosis of gastric cancer patients with bone marrow metastases. METHODS: Between September 1994 and February 2006, 39 gastric cancer patients with pathologically confirmed bone marrow dissemination were selected. RESULTS: The majority of the patients showed younger age, poorly differentiated adenocarcinoma or signet ring cell carcinoma, thrombocytopenia, anemia, elevated lactate dehydrogenase and alkaline phosphatase. Poor prognostic factors for survival were serum sodium < or =133 mmol/l [relative risk (RR) 4.57; 95% CI 1.99-10.52; p < 0.001], the presence of lung metastasis (RR 3.47; 95% CI 1.48-8.15; p = 0.007) and the presence of peritoneal seeding (RR 2.17; 95% CI 1.06-4.43; p = 0.036). Median survival durations after bone marrow metastases for patients without any adverse factors (n = 19, 48.7%) and those with 1-3 adverse factors (n = 20, 51.3%) were 67 and 23 days, respectively (p = 0.013). Patients without any adverse factors did benefit from palliative chemotherapy (p = 0.048). CONCLUSION: We suggest that gastric cancer patients with bone marrow metastases should receive more tailored therapies according to different risk factors in order to enhance survival. PMID- 18418013 TI - Safety and activity of sorafenib in different histotypes of advanced renal cell carcinoma. AB - BACKGROUND: The aim of our study was to evaluate the efficacy and safety in unresectable or advanced renal carcinoma treated with sorafenib, in a situation closely similar to the everyday medical practice. PATIENTS AND METHODS: One hundred and thirty-six patients have been treated with 400 mg b.i.d. of sorafenib administered orally until disease progression or unacceptable toxicity. They were either previously untreated or relapsed after one or more previous treatments with systemic therapy. Most of them had clear cell renal carcinoma (RCC), but other histological types such as papillary, chromophobe, Bellini ducts, sarcomatoid and mixed forms were also represented. RESULTS: Overall disease control of 70.6% was achieved with 7.9% of partial remissions. Response was observed in the majority of patients with RCC, but also in some patients with non clear cell RCC. Safety was acceptable, with the most common adverse events consisting of hand-foot skin reaction, cutaneous rash, diarrhoea, fatigue and hypertension. CONCLUSIONS: The results confirm previous ones reported in the literature concerning the efficacy and the safety of sorafenib as second-line treatment in patients with RCC. In addition, they disclose the hypothesis that sorafenib could be effective also in patients who underwent multiple previous treatments and in those with histology different from clear cells. PMID- 18418014 TI - Acute tumor lysis syndrome during oral fludarabine treatment for CLL--a rare event that might be observed more frequently in the future. PMID- 18418015 TI - Antiangiogenic metronomic chemotherapy. PMID- 18418016 TI - Preoperative short-course radiotherapy in locally advanced rectal cancer: the attitude on how to use radiotherapy in rectal cancer is changing. PMID- 18418017 TI - Response to preoperative short-course radiotherapy in locally advanced rectal cancer: value of f-fluorodeoxyglucose positron emission tomography. AB - BACKGROUND: 18F-fluorodeoxyglucose positron emission tomography (FDG-PET) is increasingly used to monitor response to multimodality cancer treatment. Preoperative short-course radiotherapy (SCRT) has been shown to improve outcome in locally advanced rectal cancer (LARC). We evaluated the value of FDGPET in monitoring response to SCRT. PATIENTS AND METHODS: 32 patients with LARC scheduled for SCRT underwent FDG-PET, magnetic resonance imaging and endorectal ultrasound both before and after SCRT. A comparison group of 16 patients, undergoing preoperative radiochemotherapy (RCT), received an extra FDG-PET 17 days after starting RCT (effective dose of 30.6 Gy). For response assessment, FDG uptake (maximum standardized uptake value, SUVmax), percent SUVmax difference, tumor regression grade (TRG), and UICC downstaging and downsizing were considered. RESULTS: SCRT resulted in a significant median SUVmax reduction of 39.3%. Median SUVmax reduction did differ between SCRT and the early scans after 17 days of RCT (39.3 vs. 50.8%). Downstaging after SCRT was observed in 16/32, downsizing in 7/32 patients. There was no correlation between SUVmax reduction and downstaging/ downsizing or TRG. CONCLUSION: For the first time, we were able to demonstrate functional response after SCRT. In contrast to morphological imaging and histopathology, our results indicate that FDG-PET can monitor early effects of SCRT. Further followup is warranted to establish SUVmax reduction as a surrogate marker for the response of SCRT. PMID- 18418018 TI - EGFR/KRAS mutations and gefitinib therapy in Chinese NSCLC patients. AB - BACKGROUND: For gefitinib treatment for non-small cell lung cancer (NSCLC), KRAS mutations reportedly behave as a resistance marker, and the epidermal growth factor receptor (EGFR) as a responsive marker. It is known that Asians and Caucasians have different responses to gefitinib. We investigated the KRAS and EGFR mutation status in a group of Chinese patients with advanced NSCLC who were treated with gefitinib after a failed chemotherapy. MATERIAL AND METHODS: Genomic DNA extracted from tumor specimens of 24 patients with advanced NSCLC, who failed at least 1 prior platinum-based chemotherapy regimen before gefitinib treatment, was subjected to nested polymerase chain reaction (PCR) to amplify codons 12, 13, 59, and 61 of the KRAS gene and exons 18-21 of the EGFR gene for direct sequencing. RESULTS: For the 24 patients, no KRAS gene mutation was found. 15 patients (62.5%, 15/24) harbored EGFR mutations which included deletion mutations in exon 19 and missense mutations in exon 21. CONCLUSION: KRAS mutation may occur at a very low frequency in Chinese NSCLC patients regardless of pathology, smoking status, or gender. Unlike EGFR, the low incidence of KRAS mutations may undermine its role in predicting the clinical response to EGFR tyrosine kinase inhibitors. PMID- 18418019 TI - Ewing's sarcoma and peripheral primitive neuroectodermal tumor in adults: different features of a rare neoplasm. AB - INTRODUCTION: In adults, peripheral primitive neuroectodermal tumors (pPNETs) represent a rare and heterogeneous group of neoplasms exhibiting neuronal and glial differentiation. PATIENTS AND METHODS: We present the clinicopathologic features of four examples of the Ewing's sarcoma (EWS)/pPNET group in adults. Hematoxylin and eosin staining, immunohistochemical and molecular studies were reviewed in every case. Immunohistochemical stains were performed on formalin fixed, paraffin-embedded sections, molecular studies were done using fluorescence in situ hybridization (FISH). RESULTS: Three patients presented with tumors of the thoracopulmonary region, one patient showed EWS of the soft tissue. Microscopically, tumor tissue was composed of round, small, blue cells with fine granular chromatin texture and inconspicuous nucleoli. Mitotic figures and rosettes were present. Tumor cells strongly coexpressed CD99 and vimentin, but due to technical reasons t(22q12) translocation studies proved the presumptive diagnosis of EWS/pPNET in one case only. Despite similar multimodality treatment survival time ranged from 6 to 42 months, two patients were alive at the time of reporting. CONCLUSIONS: As tumors of the EWS/pPNET family behave aggressively, rapid diagnosis is warranted. Since diagnosis of EWS/pPNET requires ancillary studies, it is necessary to consider it even in adult patients. PMID- 18418020 TI - A pilot study of docetaxel and trofosfamide as second-line 'metronomic' chemotherapy in the treatment of metastatic non-small cell lung cancer (NSCLC). AB - BACKGROUND: The aim of this pilot study was to evaluate the efficacy and safety of a chemotherapy containing docetaxel and oral trofosfamide as a 'metronomic' secondline treatment of patients with metastatic non-small cell lung cancer (NSCLC). PATIENTS AND METHODS: 21 patients with stage IV disease NSCLC who had progressed under first-line chemotherapy were enrolled. Previous chemotherapy was platinum-based in 15 patients (71.4%), whereas 6 patients (28.6%) had received platinum-free combination chemotherapy. Patients received docetaxel 25 mg/m(2) on days 1, 8, and 15 every 4 weeks plus trofosfamide 50 mg per day. RESULTS: A total of 62 chemotherapy cycles were administered. The median number of cycles per patient was 3. The overall response rate to chemotherapy was 19%, median overall survival was 6.9 months, the median progression-free survival 2.9 months, the 1 year survival rate 28.6%, and the 2-year survival rate 7.1%. No grade IV toxicity was observed. CONCLUSIONS: Our results suggest that the combination of docetaxel and trofosfamide in a metronomic schedule is active and well tolerable as second line therapy in patients with metastatic NSCLC. The concept of metronomic chemotherapy promises to be a valuable addition to the existing treatment options in NSCLC and warrants further investigation in phase III studies. PMID- 18418021 TI - Extraneural spread of glioblastoma--report of two cases. AB - BACKGROUND: Glioblastoma, a highly aggressive tumor, accounts for the majority of all primary brain tumors in adults. Despite a destructive local growth pattern, extraneural spread of these tumors is extremely rare. CASE 1: We describe the case of a 58-year-old man with glioblastoma, in whom an epidural mass was diagnosed 5 months after initial local therapy of the brain. A positron emission tomography (PET) scan revealed multiple metastases in the lungs, in the retroperitoneum, and in the left trochanter minor region. A soft tissue swelling of the right thigh was histologically proven to be metastatic tissue from the primary glioblastoma. The patient died 11 months after initial diagnosis. CASE 2: A 47-year-old woman with recurrent glioblastoma had a long lasting complete response to chemotherapy. 2 years after initial diagnosis she presented with a pleural mass which was a metastasis of the formerly diagnosed glioblastoma. CONCLUSION: Although systemic metastases in glioblastoma are rare, different organs can be involved. PMID- 18418022 TI - Acute tumor lysis syndrome during oral fludarabine treatment for chronic lymphocytic leukemia. Role of treatment with rasburicase. AB - INTRODUCTION: Oral fludarabine is the first oral formulation of a purine analogue available for clinical use which has produced encouraging results in patients with B-cell chronic lymphocytic leukemia (CLL). It has a similar tolerability profile to that of the intravenous formulation. Acute tumor lysis syndrome (TLS) induced by fludarabine appears to be a fairly unusual complication during the treatment of CLL. CASE REPORT: The present report describes the case of a 78-year old man who developed acute renal failure associated with TLS following the first course of oral fludarabine. Laboratory investigations fulfilled the TLS typical criteria, including hyperkalemia, marked hyperuricemia, hyperphosphatemia, hypocalcemia, and acute renal failure. Despite the many documented side effects of oral fludarabine, there is only one similar case of oral fludarabine-induced TLS reported to date. Conventional therapy with short-term supportive hemodialysis and a short treatment of rasburicase for 2 days achieved a complete recovery of renal function and a decrease of the urate level to within the normal range. CONCLUSIONS: Clinicians should be alert to this potentially life threatening metabolic emergency among CLL patients treated with oral fludarabine. PMID- 18418023 TI - Ovarian carcinoma with simultaneous breast and rectum metastases. AB - BACKGROUND: Metastatic involvement of the breast and the rectum from ovarian carcinoma are very rare events. CASE REPORT: We report a case of ovarian carcinoma with metastasis to the breast and rectum simultaneously, 6 years after initial diagnosis. RESULTS: Morphologic and immunohistochemical findings from pathologic samples of all involved sites confirmed the ovarian origin, which spared the patient unnecessary breast and rectal surgery. To our knowledge, this is the first case of ovarian carcinoma with simultaneous metastases to the breast and rectum reported to date. CONCLUSION: Accurate differential diagnosis from primary breast and rectal carcinoma is very important because the prognosis and treatment differ significantly. PMID- 18418024 TI - Flumazenil in the treatment of dizziness and vegetative symptoms after intravenous infusion of paclitaxel. PMID- 18418025 TI - Irrational healers? PMID- 18418026 TI - Critical thinking about adverse drug effects: lessons from the psychology of risk and medical decision-making for clinical psychopharmacology. AB - Systematic biases in decision-making have been well characterized in medical and nonmedical fields but mostly ignored in clinical psychopharmacology. The purpose of this paper is to sensitize clinicians who prescribe psychiatric drugs to the issues of the psychology of risk, especially as they pertain to the risk of side effects. Specifically, the present analysis focuses on heuristic organization and framing effects that create cognitive biases in medical practice. Our purpose is to increase the awareness of how pharmaceutical companies may influence physicians by framing the risk of medication side effects to favor their products. PMID- 18418027 TI - Management of factitious disorders: a systematic review. AB - BACKGROUND: The literature regarding the management of factitious disorder (FD) is diverse and generally of case reports or case series. To date there has been no systematic review of the effectiveness of management techniques. METHODS: Systematic review of all evidence reporting the management and subsequent outcome in FD. Data were extracted and outcomes were assessed using an adaptation of the Global Improvement Scale. Results were analysed by parametric statistical tests; a meta-analysis was not possible. RESULTS: Thirty-two case reports and 13 case series were eligible for inclusion. Analysis of the case reports found no significant difference in outcomes between confrontational and non confrontational approaches [t(29) = 0.72, p = 0.48], between treatment with psychotherapy compared to no psychotherapy [t(30) = 0.69, p = 0.48], and when psychiatric medication had been prescribed compared with not [t(30) = 0.35, p = 0.73]. A trend was observed that a longer length of treatment lead to better outcomes, but this was not significant [F(5, 26) = 1.17, p = 0.35]. The consecutive case series demonstrated that many FD sufferers were not engaged in treatment and were lost to follow-up but did not provide any strong evidence regarding the effectiveness of different management approaches. CONCLUSIONS: There is an absence of sufficient robust research to determine the effectiveness of any management technique for FD. The establishment of a central reporting register to facilitate the development of evidence-based guidelines is recommended. PMID- 18418028 TI - Systematic review of the effects of shared decision-making on patient satisfaction, treatment adherence and health status. AB - BACKGROUND: In the last decade, the clinician-patient relationship has become more of a partnership. There is growing interest in shared decision-making (SDM) in which the clinician and patient go through all phases of the decision-making process together, share treatment preferences, and reach an agreement on treatment choice. The purpose of this review is to determine the extent, quality, and consistency of the evidence about the effectiveness of SDM. METHOD: This is a systematic review of randomised controlled trials (RCTs) comparing SDM interventions with non-SDM interventions. Eleven RCTs met the required criteria, and were included in this review. RESULTS: The methodological quality of the studies included in this review was high overall. Five RCTs showed no difference between SDM and control, one RCT showed no short-term effects but showed positive longer-term effects, and five RCTs reported a positive effect of SDM on outcome measures. The two studies included of people with mental healthcare problems reported a positive effect of SDM. CONCLUSIONS: Despite the considerable interest in applying SDM clinically, little research regarding its effectiveness has been done to date. It has been argued that SDM is particularly suitable for long-term decisions, especially in the context of a chronic illness, and when the intervention contains more than one session. Our results show that under such circumstances, SDM can be an effective method of reaching a treatment agreement. Evidence for the effectiveness of SDM in the context of other types of decisions, or in general, is still inconclusive. Future studies of SDM should probably focus on long-term decisions. PMID- 18418029 TI - Behavioral conditioning of antihistamine effects in patients with allergic rhinitis. AB - BACKGROUND: Allergic symptoms can be induced by behavioral conditioning. However, the conditionability of antiallergic effects has not yet been studied. Thus, we investigated whether the effects of a histamine 1 (H(1)) receptor antagonist are inducible in patients suffering from house-dust mite allergy using a behavioral conditioning procedure. METHODS: During the association phase, 30 patients with allergic house-dust mite rhinitis received a novel-tasting drink once daily, followed by a standard dose of the H(1) receptor antagonist, desloratadine, on 5 consecutive days. After 9 days of drug washout, the evocation trial commenced: 10 patients received water together with an identically looking placebo pill (water group), 11 patients were re-exposed to the novel-tasting drink and received a placebo pill [conditioned stimulus (CS); CS group] and 9 patients received water and desloratadine (drug group). RESULTS: During the association phase, desloratadine treatment decreased the subjective total symptom scores, attenuated the effects of the skin prick test for histamine and reduced basophil activation ex vivo in all groups. During the evocation trial, the water group, in which subjects were not re-exposed to the gustatory stimulus, showed a reduction in subjective total symptom scores and skin prick test results, but no inhibition of basophil activation. In contrast, re-exposure to the novel-tasting drink decreased basophil activation, the skin prick test result and the subjective symptom score in the CS group to a degree that was similar to the effects of desloratadine in the drug group. CONCLUSIONS: These data show that behaviorally conditioned effects are not only able to relieve subjective rhinitis symptoms and allergic skin reactions, but also to induce changes in effector immune functions. PMID- 18418030 TI - Disruption of aryl hydrocarbon receptor (AhR) induces regression of the seminal vesicle in aged male mice. AB - The aryl hydrocarbon receptor (AhR) is a ligand-activated transcription factor that mediates diverse dioxin toxicities. Despite mediating the adverse effects, the AhR gene is conserved among animal species, suggesting important physiological functions for AhR. In fact, a recent study revealed that AhR has an intrinsic function in female reproduction, though its role in male reproduction is largely unknown. In this study, we show age-dependent regression of the seminal vesicles, probably together with the coagulating gland, in AhR(-/-) male mice. Knockout mice had abnormal vaginal plugs, low sperm counts in the epididymis, and low fertility. Moreover, serum testosterone concentrations and expression of steroidogenic 3betahydroxysteroiddehydrogenase (3betaHsd) and steroidogenic acute regulatory protein (StAR) in testicular Leydig cells were decreased in AhR(-/-) males. Taken together, our results suggest that impaired testosterone synthesis in aged mice induces regression of seminal vesicles and the coagulating glands. Such tissue disappearance likely resulted in abnormal vaginal plug formation, and eventually in low fertility. Together with previous findings demonstrating AhR function in female reproduction, AhR has essential functions in animal reproduction in both sexes. PMID- 18418031 TI - Spatial distribution of histone isoforms on the bovine active and inactive X chromosomes. AB - The inactive X chromosome (Xi) in female mammals serves as an important model for studying the role of histone isoforms in directing specific nuclear processes leading to inherited differences in transcription. In the present study, we investigated the distribution of some histone isoforms known to be involved in the process of human X inactivation on their bovine counterparts. To ascertain the identity of active and inactive X chromosome, their distribution was investigated on the X chromosomes in a cell line derived from a bovine female carrying an X;autosome translocation rcp(Xp+;23q-) which allowed the recognition of the maternal (translocated) and paternal (normal) X chromosome. The distribution patterns of histone H3 trimethylated at lysine 9 (H3K9me3) and trimethylated at lysine 27 (H3K27me3), and histone macroH2A1 and macroH2A2 (isoforms specific to heterochromatin) were determined by immunocytochemistry and compared to the temporal pattern of replication using BrdU pulse labeling prior to staining. Immunostaining revealed that H3K9me3, H3K27me3, and macroH2A1 are preferentially concentrated on the Xi, whereas the histone variant macroH2A2 is not a marker for this chromosome. H3K9me3, H3K27me3, and macroH2A1 were consistently located in bands along the Xi, while H3K9me3, macroH2A1 and macroH2A2 localized in the pericentromeric regions of the autosomes. H3K27me3 identified two intense bands on the Xi at Xp22 and Xq31, representing the early replication regions of the chromosome. H3K27me3 and macroH2A1 overlapped in the Xq31 region. It was concluded that different heterochromatin regions on the bovine inactive X chromosome can be identified by their histone isoform composition. PMID- 18418032 TI - XX/XY chimerism in cattle: clinical and cytogenetic studies. AB - The freemartin condition represents the most frequent form of intersexuality found in cattle, and occasionally in other species. Freemartinism arises when vascular connections occur between placentae of developing heterosexual twin foetuses, XX/XY chimerism develops, and ultimately there is masculinisation of the female tubular reproductive tract to varying degrees. The aim of this work was to report the clinical and cytogenetic studies performed in 28 cattle co twins, 24 of which were cytogenetically chimeric (2n = 60, XX/XY), raised in the region of Campania (southern Italy). Clinical findings of the 16 freemartin females examined varied greatly, from a more female phenotype (normal body conformation with the presence of a blind-ending vagina and primordial ovarian and uterus structures) to a nearly male phenotype (body conformations with male traits and presence of primordial prepuce, penis and testicles). The 8 freemartin males, in spite of the presence of XX cells, had a normal body conformation and external genitalia and some of them were fertile. In addition to cytogenetic diagnosis we also verified chromosome fragility by testing for chromosome aberration (CA: aneuploidy, gaps, chromatid breaks, chromosome breaks and fragments) and sister chromatid exchange (SCE). Freemartins showed a higher percentage of aneuploid cells and significant statistical differences in mean values of gaps, chromatid breaks and chromosome breaks when compared with control animals. To our knowledge, this is the first time that chromosome instability has been evaluated by analyses of CA and SCE in freemartin cattle. PMID- 18418033 TI - Subcellular redistribution of the mitochondrial PG2 epitope during development from cleavage to primordial germ cell formation in the rabbit embryo. AB - Mitochondria are central players in diverse cellular functions and their efficient functioning has dramatic impact on embryonic development. Apparently, proliferation and transmission of well functioning mitochondria to the next generation require ingeniously adapted mechanisms, one of which, the 'mitochondrial bottleneck', is thought to occur early in mammalian development during primordial germ cell (PGC) specification. We used an antibody directed against the mitochondrial PG2 epitope, a reliable marker of primordial and adult female germ cells to monitor mitochondrial differentiation in the early rabbit embryo. Early development shows the PG2 epitope either tightly mitochondria associated (zygote) or diffusely distributed throughout cytoplasm (cleavage stages). Mitochondrial colocalization of the PG2 epitope is regained in the early blastocyst but expression is then retained by the hypoblast and epiblast only, with the epiblast, although the forerunner of PGCs, showing weak and diffuse labeling only. At gastrulation, hypoblast cells lose PG2 expression but intensive PG2 labeling is found again on all mitochondria in the first PGCs and reveals the number of mitochondria to be in the range of 50 to 100 per PGC at this stage. The results highlight the dynamics of PG2 expression during early development and the usefulness of the epitope for testing the bottleneck theory. PMID- 18418034 TI - Genetics of sex determination in tilapiine species. AB - We identified DNA markers linked to sex determining genes in six closely related species of tilapiine fishes. The mode of sex determination differed among species. In Oreochromis karongae and Tilapia mariae the sex-determining locus is on linkage group (LG) 3 and the female is heterogametic (WZ-ZZ system). In O. niloticus and T. zillii the sex-determining locus is on LG1 and the male is heterogametic (XX-XY system). A more complex pattern was observed in O. aureus and O. mossambicus, in which markers on both LG1 and LG3 were associated with sex. We found evidence for sex-linked lethal effects on LG1, as well as interactions between loci in the two linkage groups. Comparison of genetic and physical maps demonstrated a broad region of recombination suppression harboring the sex-determining locus on LG3. Sex-specific recombination suppression was found in the female heterogametic sex. Sequence analysis showed the accumulation of repetitive elements in this region. Phylogenetic analysis suggests that at least two transitions in the mode of sex determination have occurred in this clade. This variation in sex determination mechanisms among closely related species makes tilapias an excellent model system for studying the evolution of sex chromosomes in vertebrates. PMID- 18418035 TI - Telomeres: the silence is broken. AB - The ends of linear eukaryotic chromosomes, telomeres, distinguish natural chromosome ends from DNA double stranded breaks and thus promote genome stability. Telomeres comprise a repetitive DNA skeleton, which is wrapped in specific protein complexes. Recent data indicate that an additional building block of telomeres is RNA and that the longstanding idea that telomeres are silent genomic regions needs to be overturned. Mammalian telomeres are indeed transcribed into RNA molecules, which remain associated with telomeric chromatin, suggesting RNA-mediated mechanisms in organizing telomere architecture. PMID- 18418036 TI - Is NHEJ a tumor suppressor or an aging suppressor? AB - Tumor suppressors are longevity assurance genes that ensure early life fitness. Genes are defined as tumor suppressors if their mutation predisposes the animal to cancer (a phenotype-based definition). Tumor suppressors fall into two categories: caretakers and gatekeepers. Caretakers suppress cancer by repairing damaged DNA while gatekeepers suppress cancer by halting the cell cycle long enough to repair damaged DNA. If the damage is irreparable, gatekeepers induce either apoptosis or senescence. These responses are deleterious to the cell but protect the organism. p53 is the best-known gatekeeper because it is mutated in over half of all cancers. Nonhomologous end joining (NHEJ) is considered a caretaker since it repairs DNA double-strand breaks that would otherwise lead to gross chromosomal rearrangements (GCRs). NHEJ-mutant mice display increased GCRs, but without increased cancer. Instead these mice show early aging. This commentary focuses on the role NHEJ has on aging and cancer. I propose that NHEJ evolved to reduce GCRs and moderate gatekeeper responses that would otherwise cause early aging. Furthermore, NHEJ did not evolve to suppress tumors and any observed tumor suppression is merely circumstantial to unnatural laboratory conditions coupled with human bias that favors defining all DNA repair pathways as caretakers. PMID- 18418037 TI - Butyrate and Wnt signaling: a possible solution to the puzzle of dietary fiber and colon cancer risk? AB - Studies on the protective role of dietary fiber and its breakdown product butyrate against colorectal cancer (CRC) have yielded inconsistent findings. We have reported that butyrate treatment of CRC cells in vitro modulates canonical Wnt signaling, a pathway which is constitutively activated in the majority of CRCs. Analyses of ten human CRC cell lines exposed to butyrate have established that the levels of apoptosis in these cells are dependent upon the fold induction of canonical Wnt transcriptional activity. It is likely that the observed variability in the levels of induced Wnt activity and apoptosis in CRC cells in vitro reflects the existence of different CRC subtypes in vivo. The existence of CRC subtypes, individual- and population-specific variation in butyrate producing colonic microflora, and the time at which the colorectal lesions (early vs. late stage) are exposed to fiber/butyrate are all factors that may influence the protective role of fiber against CRC. We discuss the evidence by which these factors influence the effects of fiber on colonic tumorigenesis and outline experimental approaches for testing these hypotheses. PMID- 18418038 TI - Shedding light on an old mystery: thalidomide suppresses survival pathways to induce limb defects. AB - Many hypotheses have been proposed to explain the molecular mechanism of thalidomide teratogenicity, in particular regarding to limb defects. Most experimental evidence in vivo has been provided for a model that suggests the generation of oxidative stress by thalidomide with subsequent downregulation of Wnt and Akt survival pathways. As a consequence apoptosis is induced during early embryonic limb development resulting in limb truncations. Here we summarize and discuss the relevant data supporting this hypothesis. We extend this model by presenting new data demonstrating an involvement of the transcription factors Tbx5 and Sall4 in thalidomide-induced molecular pathology. Finally, we discuss a possible participation of other stress-responsive and/or pro-apoptotic transcription factors in the mechanism of thalidomide teratogenicity. PMID- 18418039 TI - Cdc25A promotes G2/M transition in oocytes. PMID- 18418040 TI - c-Met inhibitors with different binding modes: two is better than one. AB - Primary and acquired resistance to kinase inhibitors due to pre-existing mutations of the target or to mutations that arise as a result of selection by therapy is now a common theme in cancer patients treated with these drugs. Different classes of inhibitors for the same target have been successful in overcoming, at least temporarily, these resistance mechanisms because of their ability to interact with the mutated receptor. Therefore, having different classes of inhibitors for a given target might offer more treatment options for cancer patients. c-Met inhibitors are emerging as potentially important new cancer drugs and profiling these agents against several mutant receptors has begun. We have recently identified c-Met inhibitors that are active against wild type and mutated c-Met variants. X-ray crystallography revealed that this class of inhibitors binds c-Met very differently than another c-Met inhibitor that shows primary resistance to some c-Met mutants. Our results suggested that it is possible to identify c-Met inhibitors that will be active against a range of c Met mutations. PMID- 18418041 TI - Epigenetic inheritance of cell differentiation status. AB - Epigenetic modifications influence gene expression pattern and provide a unique signature of a cell differentiation status. Without external stimuli or signalling events, this cell identity remains stable and unlikely to change over many cell divisions. The epigenetic signature of a particular cell fate therefore needs to be replicated faithfully in daughter cells; otherwise a cell lineage cannot be maintained. However, the mechanism of transmission of cellular memory from mother to daughter cells remains unclear. It has been suggested that the inheritance of an active or silent gene state involves different kinds of epigenetic mechanisms, e.g. DNA methylation, histone modifications, replacement of histone variants, Polycomb group (PcG) and Trithorax group (TrxG) proteins. Emerging evidence supports the role of histone variant H3.3 in maintaining an active gene status and in remodelling nucleosomal composition. Here we discuss some recent findings on the propagation of epigenetic memory and propose a model for the inheritance of an active gene state through the interaction of H3.3 with other epigenetic components. PMID- 18418042 TI - Hypoxia-inducible factor signaling in the development of tissue fibrosis. AB - Capillary rarefaction is a hallmark of fibrotic diseases and results in reduced blood perfusion and oxygen delivery. In the kidney, tubulointerstitial fibrosis, which leads to the destruction of renal tissue and the irreversible loss of kidney function, is associated with hypoxia and the activation of Hypoxia Inducible-Factor (HIF) signaling. HIF-1 and HIF-2 are basic-helix-loop-helix transcription factors that allow cells to survive in a low oxygen environment by regulating energy metabolism, vascular remodeling, erythropoiesis, cellular proliferation and apoptosis. Recent studies suggest that HIF activation promotes epithelial to mesenchymal transition (EMT) and renal fibrogenesis. These findings raise the possibility that the spectrum of HIF activated biological responses to hypoxic stress may differ under conditions of acute and chronic hypoxia. Here we discuss the role of HIF signaling in the pathogenesis and progression of chronic kidney disease. PMID- 18418043 TI - Insights into the oncogenic effects of PIK3CA mutations from the structure of p110alpha/p85alpha. AB - Phosphatidylinositide-3-kinases (PI3K) initiate a number of signaling pathways by recruiting other kinases, such as Akt, to the plasma membrane. One of the isoforms, PI3Kalpha, is an oncogene frequently mutated in several cancer types. These mutations increase PI3K kinase activity, leading to increased cell survival, cell motility, cell metabolism, and cell cycle progression. The structure of the complex between the catalytic subunit of PI3Kalpha, p110alpha, and a portion of its regulatory subunit, p85alpha reveals that the majority of the oncogenic mutations occur at the interfaces between p110 domains and between p110 and p85 domains. At these positions, mutations disrupt interactions resulting in changes in the kinase domain that may increase enzymatic activity. The structure also suggests that interaction with the membrane is mediated by one of the p85 domains (iSH2). These findings may provide novel structural loci for the design of new anti-cancer drugs. PMID- 18418044 TI - Wnt signaling and adipocyte lineage commitment. AB - Obesity is characterized by an increase in the number mature fat cells. These nascent adipocytes are derived from preadipocytes, which in turn are derived from mesenchymal stem cells (MSCs). Since little is known about the mechanisms controlling the commitment of MSCs into preadipocytes, this early event in adipogenesis was further investigated. C3H10T1/2 cells (10T1/2 cells) were employed as a MSC model and a committed A33 preadipocyte cell line derived from these cells served as a model of preadipocytes. Microarray technology was used to identify genes that are differentially expressed in pluripotent 10T1/2 cells when compared with A33 preadipocytes. Several key genes of the Wnt signaling pathway were differentially expressed between 10T1/2 and A33 cells as demonstrated by microarray and quantitative real-time RT-PCR analyses. Of particular interest, R spondins-2 and -3, newly described molecules that activate the canonical Wnt signaling pathway, are markedly upregulated in proliferating A33 cells compared to 10T1/2 cells. Consistent with these findings beta-catenin accumulates in the nuclei of proliferating A33 cells, but not 10T1/2 cells. In addition, several members of the Lef/Tcf family of transcription factors involved in Wnt signaling are also differentially expressed between 10T1/2 and A33 cells. These and other findings indicate that activation of Wnt signaling is an early event in adipogenesis. PMID- 18418045 TI - Irreversible chromosome damage accumulates rapidly in the absence of ATM kinase activity. AB - Mutations in the ATM kinase cause the neurodegenerative disorder ataxia telangiectasia (A-T) and affected individuals are exquisitely radiation-sensitive and cancer-prone. Cells derived from A-T individuals contain chromosome aberrations and exhibit profound cellular radiosensitivity. ATM is an apical kinase critical for the activation of cell cycle checkpoints and the induction of apoptosis in irradiated cells. However, defects in these pathways are insufficient to account for the chromosomal instability seen in A-T cells. We show here that the small molecule KU55933 can be used as a "molecular switch" to selectively and transiently inhibit ATM kinase activity in cells. We subsequently show that the cellular radiosensitization seen when ATM kinase activity is inhibited for one hour following exposure to gamma-rays, accounts for over 70% of the total cellular radiosensitization seen when ATM kinase activity is inhibited for 17 h. Finally, we show that inhibition of ATM kinase activity for one hour following exposure to irradiation doubles the number of chromosome aberrations occurring in late-S- and G(2)-, but not M-phase, cells. These observations are unexpected and suggest that irreversible chromosome damage accumulates very rapidly when ATM kinase activity is transiently inhibited following irradiation. We propose that we have revealed an essential, yet previously undescribed, role for ATM kinase in suppressing chromosomal instability. PMID- 18418046 TI - ESCRT functions in autophagy and associated disease. AB - Mutations in the endosomal sorting complexes required for transport (ESCRT)-III subunit CHMP2B are associated with frontotemporal dementia (FTD) and amyotrophic lateral sclerosis (ALS), both human neurodegenerative diseases characterized by accumulation of ubiquitinated proteins aggregates in affected neurons. The ESCRT proteins are known to be involved in diverse cellular processes such as mRNA transport, cytokinesis, transcriptional regulation and sorting of transmembrane proteins into the inner vesicles of the multivesicular body (MVB) during endocytosis. It was until recently not clear how ESCRT function may be involved in neurodegeneration. New findings in mammalian cells and in Drosophila melanogaster show that functional ESCRTs are required for efficient fusion of autophagic vesicles with the endocytic pathway and for degradation of autophagic cargo. Moreover, defective ESCRT function led to the accumulation of cytoplasmic protein aggregates containing ubiquitin, p62/Sequestosome-1 and TAR DNA binding protein 43 (TDP-43). Using cellular and Drosophila models for Huntington's disease it was also shown that reduced ESCRT levels inhibit clearance of expanded polyglutamine aggregates and aggravate their neurotoxic effect. These data indicate that efficient autophagic degradation requires functional MVBs and provides a possible explanation to the observed neurodegenerative phenotype seen in patients with CHMP2B mutations. PMID- 18418047 TI - A role of HAUSP in tumor suppression in a human colon carcinoma xenograft model. AB - The protease HAUSP is a critical component of the p53-Mdm2 pathway and acts as a specific deubiquitinase for both p53 and Mdm2 and thus is important for p53 regulation. In knock-down and knock-out cellular systems it was observed that ablation of HAUSP induces profound stabilization of p53 due to enhanced degradation of Mdm2. Thus, inhibiting HAUSP by small compound interference has been proposed as a rational therapeutic strategy to activate p53 in p53 wild type tumors. However, HAUSP-mediated effects in the p53-Mdm2 axis are highly complex and non-linear and to date the role of HAUSP in tumor suppression in vivo remains unexplored. Here we investigate the effect of HAUSP up and downregulation on cell proliferation, apoptosis and tumor growth in vitro and in a xenograft model in vivo, using an inducible isogenic human colon carcinoma cell system. Importantly, in the absence of stress, both HAUSP up and downregulation inhibit cell proliferation in vitro and tumor growth in vivo due to constitutively elevated p53 levels. Moreover, tumors with HAUSP up and downregulation respond to radiotherapy with further growth inhibition. However, HAUSP downregulation causes resistance to Camptothecin- and irradiation-induced apoptosis, which correlates with suppressed mitochondrial translocation of p53. Our data suggest that changes in HAUSP modulate tumor growth and apoptotic sensitivity in vivo. PMID- 18418048 TI - Caspase 2 is both required for p53-mediated apoptosis and downregulated by p53 in a p21-dependent manner. AB - Upon treatment with some DNA damaging agents, human H1299 tumor-derived cells expressing inducible versions of wild-type or mutant p53 with inactive transactivation domain I (p53(Q22/S23)) undergo apoptosis. In cells expressing either version of p53, caspase 2 activation is required for release of cytochrome c and cell death. Furthermore, silencing of PIDD (a factor previously shown to be required for caspase 2 activation) by siRNA suppresses apoptosis by both wild type p53 and p53(Q22/S23). Despite the finding that caspase 2 is essential for DNA damage-facilitated, p53-mediated apoptosis, induction of wild-type p53 (with or without DNA damage) resulted in a reduction of caspase 2 mRNA and protein levels. In this study we sought to provide a mechanism for the negative regulation of caspase 2 by p53 as well as provide insight as to why p53 may repress a key mediator of p53-dependent apoptosis. Mechanistically, we show that DNA binding and/or transactivation domains of p53 are crucial for mediating transrepression. Further, expression of p21 (in p53-null cells inducibly expressing p21) is sufficient to mediate repression of caspase 2. Deletion of p21 or E2F-1 not only abrogated repression of caspase 2, but also stimulated the expression of caspase 2 above basal levels, implicating the requirement for an intact p21/Rb/E2F pathway in the downregulation of caspase 2. As this p53/p21 dependent repression of caspase 2 can occur in the absence of DNA damage, caspase 2 repression does not simply seem to be a consequence of the apoptotic process. Downregulation of caspase 2 levels by p53 may help to determine cell fate by preventing cell death when unnecessary. PMID- 18418049 TI - PERK and PKR: old kinases learn new tricks. AB - Regulating gene expression is an effective way for cells to deal with various stresses. The outcome of this regulation differs with the type of stress, and can promote either cell survival or cell death depending on the severity of the injury incurred. Gene expression can be controlled at several steps, including transcription, translation and degradation. An extensively studied protein involved in translational control is the eukaryotic translation initiation factor 2 (eIF2). When eIF2 becomes phosphorylated on a specific serine residue located within the alpha (alpha) subunit, global protein synthesis is halted. This phosphorylation occurs following periods of environmental stress, and plays a significant role in the cellular response to these events. The eIF2alpha kinase family consists of four members, which are each activated in response to different stimuli. Our group has recently discovered that two members of this family, the protein kinase activated by double-stranded RNA (PKR) and the PKR like endoplasmic reticulum (ER) kinase (PERK) can also regulate the expression of specific proteins by promoting their degradation by the 26S proteasome. Specifically, we demonstrated that degradation of the cell cycle regulator cyclin D1, and the tumour suppressor p53 was promoted by PERK and PKR during periods of ER stress. This novel function may allow the eIF2alpha kinases to affect a larger number of cellular processes than previously believed. PMID- 18418050 TI - Paternal cranial irradiation induces distant bystander DNA damage in the germline and leads to epigenetic alterations in the offspring. AB - It is now well accepted that parental whole body irradiation causes transgenerational genome and epigenome instability in the offspring. The majority of human exposures to radiation, such as therapeutic and diagnostic irradiation, are localized and focused. The potential of localized body-part exposures to affect the germline and thus induce deleterious changes in the progeny has not been studied. To investigate whether or not the paternal cranial irradiation can exert deleterious changes in the protected germline, we studied the accumulation of DNA damage in the shielded testes tissue. Here we report that the localized paternal cranial irradiation results in a significant accumulation of unrepaired DNA lesions in sperm cells and leads to a profound epigenetic dysregulation in the unexposed progeny conceived a week after paternal exposure. PMID- 18418051 TI - p73-mediated transcriptional activity is negatively regulated by polo-like kinase 1. AB - Trans-activating (TA) p73 is a member of the p53 family of transcription factors and has been implicated in cell cycle regulation, apoptosis and developmental processes. Although TAp73 positively regulates an overlapping repertoire of genes regulated by p53, TAp73 has been observed to be paradoxically overexpressed in a number of tumor cell types arousing much interest in the post-translational regulation of TAp73 transcriptional activity. Here, we present novel findings that show TAp73 can interact and co-localise, with Polo-Like Kinase 1 (PLK1) and that TAp73 is phosphorylated by this kinase on Threonine-27 (Thr-27) within the TA domain. Using reporter assays and Electrophoretic Mobility Shift Assays (EMSA), our findings suggest that TAp73-mediated activation of the p21(cip/waf), 14-3-3sigma and Bax gene promoters is abrogated by expressed PLK1 for which post translational modification of TAp73 Thr-27 appears to be a key step in MCF7 cells. Thus highlighting a potential mechanism that uniquely contributes to PLK1 mediated and phosphor-dependent transcriptional deactivation of expressed TAp73. PMID- 18418052 TI - The CCN3 gene coding for an extracellular adhesion-related protein is transcriptionally activated by the p53 tumor suppressor. AB - The CCN3 protein (Nov, Nephroblastoma overexpressed) is a member of the CCN family (Cyr61, CTGF, Nov) of growth regulators and exerts antiproliferative properties. We show here that the tumor suppressor p53 transcriptionally upregulates the CCN3 gene. p53 is an important transcription factor contributing to cell cycle arrest and apoptosis after cell damage through the regulation of numerous target genes. We show that CCN3 mRNA and protein are upregulated following p53 expression. A DNA binding-deficient p53 mutant fails to regulate CCN3. CCN3 protein is located in the perinuclear space after induction and is also exported to the extracellular matrix. Furthermore, the CCN3 promoter is inducible by p53 and the response element is located in the first exon of the CCN3 gene. Chromatin immunoprecipitations show that p53 binds to the CCN3 promoter in vivo. As CCN3 was shown to inhibit cell growth, transcriptional induction by p53 may serve as an antiproliferative signal in the extracellular matrix. Furthermore, CCN3 depletion was also reported to reduce collagen type IV dependent adhesion of melanocytes. Thus, elevated levels of CCN3 protein regulated by p53 might influence cell adhesion. PMID- 18418053 TI - U-box-type ubiquitin E4 ligase, UFD2a attenuates cisplatin mediated degradation of DeltaNp63alpha. AB - DeltaNp63alpha, the dominant negative isoform of the p63 family is an essential survival factor in head and neck squamous cell carcinoma. This isoform has been shown to be down regulated in response to several DNA damaging agents, including cisplatin. But little is understood about the post-translational protein stability of DeltaNp63alpha. In this present study we demonstrate for the first time that DeltaNp63alpha physically interacts with U-box-type E4 ubiquitin ligase UFD2a. UFD2a stabilizes DeltaNp63alpha, and ubiquitylation of DeltaNp63alpha is attenuated by UFD2a both in the presence and absence of cisplatin. Ectopic expression of UFD2a increased the half-life of DeltaNp63alpha in association with a significant enhancement of the repressive transcriptional activity of DeltaNp63alpha. Downregulation of endogenous UFD2a by RNAi resulted in degradation of DeltaNp63alpha. Taken together, our current study provides an insight onto the regulation of DeltaNp63alpha protein levels in response to cisplatin and also suggests that UFD2a might play an important role in the regulation of cisplatin mediated cell death mediated by p63. PMID- 18418054 TI - Disruption of the circadian clock due to the Clock mutation has discrete effects on aging and carcinogenesis. AB - The mammalian circadian system has been implicated in the regulation of various biological processes including those involved in genotoxic stress responses and tumor suppression. Here we report that mice with the functional deficiency in circadian transcription factor CLOCK (Clock/Clock mutant mice) do not display predisposition to tumor formation both during their normal lifespan or when challenged by gamma- radiation. This phenotype is consistent with high apoptotic and low proliferation rate in lymphoid tissues of Clock mutant mice and is supported by the gene expression profiling of a number of apoptosis and cell cycle-related genes, as well as by growth inhibition of cells with CLOCK downregulation. At the same time, Clock mutant mice respond to low-dose irradiation by accelerating their aging program, and develop phenotypes that are reminiscent of those in Bmal1-deficient mice. Taken together, our results demonstrate the dichotomy in biological consequences of the disruption of the circadian clock with respect to ageing and cancer. They also highlight the existence of a complex interconnection between ageing, carcinogenesis and individual components of the circadian clock machinery. PMID- 18418055 TI - Protein complexes at the microtubule organizing center regulate bipolar spindle assembly. AB - Bipolar spindle assembly is essential to genomic stability in dividing cells. Centrosomes or spindle pole bodies duplicated earlier at G(1)/S remain adjacent until triggered at mitotic onset to become bipolar. Pole reorientation is stabilized by microtubule interdigitation but mechanistic details for bipolarity remain incomplete. To investigate the contribution of spindle pole microtubule organizing center (MTOC) proteins in bipolarity, we applied genetic, structural and molecular biochemical analysis along with timelapse microscopy. Spindle formation was followed by an in vivo growth assay with the conditional allele cut7-22(ts), encoding fission yeast mitotic Kinesin-5, essential for bipolarity. By analysis of double and triple mutant strains of MTOC alleles and cut7-22(ts) we found that stabilized microtubules or increased bundling can rescue cut7 22(ts) associated bipolarity defects. These changes to microtubule dynamics and organization occurred through two surface domains on gamma-tubulin, a helix 11 domain and an adjacent site for binding MTOC protein Alp4. We demonstrate that Kinesin-14 Pkl1, known to oppose bipolarity, can bind to gamma-tubulin at helix 11 and that mutation of either of two conserved residues in helix 11 can impair Kinesin-14 binding. Altering the Alp4/gamma-tubulin interaction, conserved residues in helix 11 or deletion of pkl1 each are sufficient to rescue bipolarity in our cut7-22(ts) strain. Our findings provide novel insights into regulation of the bipolar mechanism through the MTOC complex. PMID- 18418056 TI - Inhibition of p73 function by Pifithrin-alpha as revealed by studies in zebrafish embryos. AB - The p53 family of proteins contains two members that have been implicated in sensitization of cells and organisms to genotoxic stress, i.e., p53 itself and p73. In vitro, lack of either p53 or p73 can protect certain cell types in the adult organism against death upon exposure to DNA damaging agents. The present study was designed to assess the relative contribution of p53 to radiation resistance of an emerging vertebrate model organism, i.e., zebrafish embryos. Consistent with previous reports, suppressing p53 protein expression using antisense morpholino oligonucleotides (MOs) increased survival and reduced gross morphological alterations in zebrafish embryos exposed to ionizing radiation. By contrast, a pharmacological inhibitor of p53 function [Pifithrin-alpha(PFTalpha)] caused developmental abnormalities affecting the head, brain, eyes and kidney function and did not protect against lethal effects of ionizing radiation when administered at 3 hours post fertilization (hpf). The phenotypic abnormalities associated with PFTalpha treatment were similar to those caused by antisense MO knock down (kd) used to reduce p73 expression. PFTalpha also inhibited p73 dependent transcription of a reporter gene construct containing canonical p53 responsive promoter sequences. Notably, when administered at later stages of development (23 hpf), PFTalpha did not cause overt developmental defects but exerted radioprotective effects in zebrafish embryos. In summary, this study highlights off-target effects of the pharmacological p53 inhibitor PFTalpha related to inhibition of p73 function and essential roles of p73 at early but not later stages of zebrafish development. PMID- 18418057 TI - Distinct roles for p107 and p130 in Rb-independent cellular senescence. AB - Telomere attrition, DNA damage and constitutive mitogenic signaling can all trigger cellular senescence in normal cells and serve as a defense against tumor progression. Cancer cells may circumvent this cellular defense by acquiring genetic mutations in checkpoint proteins responsible for regulating permanent cell cycle arrest. A small family of tumor suppressor genes encoding the retinoblastoma susceptibility protein family (Rb, p107, p130) exerts a partially redundant control of entry into S phase of DNA replication and cellular proliferation. Here we report that activation of the p53-dependent DNA damage response has been found to accelerate senescence in human prostate cancer cells lacking a functional Rb protein. This novel form of irradiation-induced premature cellular senescence reinforces the notion that other Rb family members may compensate for loss of Rb protein in the DNA damage response pathway. Consistent with this hypothesis, depletion of p107 potently inhibits the irradiation-induced senescence observed in DU145 cells. In contrast, p130 depletion triggers a robust and unexpected form of premature senescence in unirradiated cells. The dominant effect of depleting both p107 and p130, in the absence of Rb, was a complete blockade of irradiation-induced cellular senescence. Onset of the p107-dependent senescence was temporally associated with p53-mediated stabilization of the cyclin-dependent kinase inhibitor p27 and decreases in c-myc and cks1 expression. These results indicate that p107 is required for initiation of accelerated cellular senescence in the absence of Rb and introduces the concept that p130 may be required to prevent the onset of terminal growth arrest in unstimulated prostate cancer cells lacking a functional Rb allele. PMID- 18418058 TI - The nucleolar phosphatase Cdc14B is dispensable for chromosome segregation and mitotic exit in human cells. AB - In yeast, the protein phosphatase Cdc14 promotes chromosome segregation, mitotic exit, and cytokinesis by reversing M-phase phosphorylations catalyzed by Cdk1. A key feature of Cdc14 regulation is its sequestration within the nucleolus, which restricts its access to potential substrates for much of the cell cycle. Mammals also possess a nucleolar Cdc14 homolog, termed Cdc14B, but its roles during mitosis and cell division remain speculative. Here we analyze Cdc14B's subcellular dynamics during mitosis and rigorously test its functional contributions to cell division through homozygous disruption of the Cdc14B locus in human somatic cells. While Cdc14B is initially released from nucleoli at the start of mitosis, the phosphatase quickly redistributes onto segregating sister chromatids during anaphase. This relocalization is mainly driven by Cdk1 inactivation, as pharmacologic inhibition of Cdk1 in prometaphase cells redirects Cdc14B onto chromosomes. However, in sharp contrast to yeast cdc14 mutants, human Cdc14B(Delta/Delta) cells were viable and lacked defects in spindle assembly, anaphase progression, mitotic exit, and cytokinesis, and continued to segregate ribosomal DNA repeats with near-normal proficiency. Our findings reveal substantial divergence in mitotic regulation between yeast and mammalian cells, as the latter possess efficient mechanisms for completing late M-phase events in the absence of a nucleolar Cdc14-related phosphatase. PMID- 18418059 TI - The Flp1/Clp1 phosphatase cooperates with HECT-type Pub1/2 protein-ubiquitin ligases in Schizosaccharomyces pombe. AB - The Schizosaccharomyces pombe Flp1p serine-threonine phosphatase is required for the degradation of the mitotic inducer Cdc25p at the end of mitosis. Cdc25p degradation prevents Cdc2p-tyrosine 15 dephosphorylation and, thus, contributes to the timely inactivation of mitotic CDK-associated kinase activity. Both RING- and HECT-type protein-ubiquitin ligases are involved in Cdc25p destabilization. Flp1p function is required for Cdc25p ubiquitination via anaphase-promoting complex/cyclosome or APC/C (RING-type) and the absence of Pub1p (HECT-type) stabilizes the mitotic inducer. In the present report, we study the functional relationship of Flp1p with Pub1p and Pub2p HECT-type-protein ubiquitin ligases. We show that Flp1p is required for the rapid degradation of Cdc25p while Pub1p is responsible for the long-term destabilization of the mitotic inducer. Accordingly, flp1 and pub1 mutants have a strong genetic interaction, correlating defects in the coordination of mitosis and cytokinesis with the stabilization of hyperactive Cdc25p. However, we also show that Flp1 and Pub2p proteins functionally interact in vivo suggesting that both proteins belong to the same regulatory network in S. pombe cells. Thus Flp1p appears to have an important role in integrating HECT- and RING-type ubiquitin ligases in cell cycle control. PMID- 18418060 TI - Amelioration of protein misfolding disease by rapamycin: translation or autophagy? AB - Rapamycin is an inhibitor of mTOR, a key component of the mTORC1 complex that controls the growth and survival of cells in response to growth factors, nutrients, energy balance and stresses. The downstream targets of mTORC1 include ribosome biogenesis, transcription, translation and macroautophagy. Recently it was proposed that rapamycin and its derivatives enhance the clearance (and/or reduce the accumulation) of mutant intracellular proteins causing proteinopathies such as tau, alpha-synuclein, ataxin-3, and full-length or fragments of huntingtin containing a polyglutamine (polyQ) expansion, by upregulating macroautophagy. We tested this proposal directly using macroautophagy-deficient fibroblasts. We found that rapamycin inhibits the aggregation of a fragment of huntingtin (exon 1) containing 97 polyQs similarly in macroautophagy-proficient (Atg5(+/+)) and macroautophagy-deficient (Atg5(-/-)) cells. These data demonstrate that autophagy is not the only mechanism by which rapamycin can alleviate the accumulation of misfolded proteins. Our data suggest that rapamycin inhibits mutant huntingtin fragment accumulation due to inhibition of protein synthesis. A model illustrates how a modest reduction in polyQ synthesis can lead to a long-lasting reduction in polyQ aggregation. We propose that several mechanisms exist by which rapamycin reduces the accumulation and potential toxicity of misfolded proteins in diseases caused by protein misfolding and aggregation. PMID- 18418061 TI - Histone H4 acetylation during UV light induced G1 and S phase arrest of the cell cycle. PMID- 18418062 TI - Cancer stem cells: Models, mechanisms and implications for improved treatment. AB - Cancer stem cell research has drawn a lot of attention recently. The importance of cancer stem cell research lies in the possibility of providing new approaches for improved understanding of cancer biology and cancer treatment. However, cancer stem cell research is still at its infancy. In this review, we will discuss the concept of cancer stem cells and the commonly used approaches for isolating or enriching cancer stem cells. We then review the common features of cancer stem cells including resistance to cancer drugs and stresses such as hypoxia and radiation. We also summarize signaling pathways which might be preferentially important for cancer stem cells. Finally, we will propose a new Yin-Yang model of cancer stem cells and discuss the possibility of developing new drugs that target cancer stem cells for improved treatment of cancer. PMID- 18418063 TI - Unraveling estrogen action in osteoporosis. AB - A decrease in estrogen levels at menopause leads to a rapid loss of bone mineral density and an increase in fracture risk. For over ten years it has been known that the beneficial effects of estrogen are due in part to the ability of estrogen to suppress osteoclastogenic cytokine production in T-cells and osteoblasts. In addition to suppressing these cytokines, estrogen has been shown to induce the apoptotic death of osteoclasts. A variety of different mechanisms have been suggested to explain the estrogen regulation of osteoclast survival. One hypothesis is that estrogen, via rapid non-genomic signaling, induces apoptosis without the need for direct binding of estrogen receptor alpha (ERalpha) to DNA. A second hypothesis proposes that estrogen-stimulation of ERalpha in osteoclasts induces the expression Fas Ligand which in turn leads to cell death via an autocrine mechanism. In contrast, recent work from our lab has led to a genomic model of estrogen action in which estrogen acts to induce ERalpha binding to transcriptional enhancers in the Fas Ligand gene leading to its upregulation in osteoblasts which through a paracrine mechanism induces apoptosis in osteoclasts. Here we will focus on these differing models of the mechanism of estrogen-mediated osteoclast apoptosis. PMID- 18418064 TI - Controlled rereplication at DNA replication origins. AB - No-more-than-once per cell cycle initiation of DNA replication is a golden rule to maintain genome stability and guarantee cell survival. In our recent work we discovered that small genome fragments of about 200 bp are repeatedly synthesised at human DNA replication origins at the time of origin firing during S phase in normal cells. Rereplicated DNA fragments coincide physical and temporarily with replication origin activity, implying that their generation is intimately associated with the initiation of DNA replication. This unexpected finding may provide a new framework to study replication origin regulation. PMID- 18418065 TI - Control of pancreatic beta-cell fate by insulin signaling: The sweet spot hypothesis. AB - Diabetes results from an absolute or relative deficiency in functional pancreatic beta-cell mass. Over the past few years, there has been renewed interest in the role of insulin itself in the regulation of beta-cell fate. Numerous animal models point to a critical role for beta-cell insulin signaling in the survival and proliferation of pancreatic beta-cells. In the present article, we review new studies that elucidate the mechanism by which insulin exerts anti-apoptotic and pro-mitogenic effects on beta-cells. In particular, we highlight the emerging role for Raf-1 kinase in autocrine insulin signaling and beta-cell fate decisions. We also discuss provocative evidence that the relationship between the dose of insulin and the birth and death of beta-cells is not linear. We propose a new hypothesis based on these findings, called the 'sweet spot' hypothesis, that can explain how both upward and downward deviations from normal levels of autocrine/paracrine insulin signaling might play an important role in the pathogenesis of type 1 diabetes and type 2 diabetes. We also highlight the key experiments that are required to further test this hypothesis. PMID- 18418066 TI - Explaining the preponderance of Kras mutations in human cancer: An isoform specific function in stem cell expansion. AB - Mutationally activated forms of the three closely related Ras isoforms, Kras, Hras and Nras can each exert oncogenic activity, and activated alleles arise in a variety of human cancers. However, mutant Kras is, by far, the most frequently observed Ras isoform in cancer, and is most frequently detected in tumors derived from endodermal tissues, including pancreas, lung and colon. We have recently reported findings that may explain this. We observed that activated Kras, but not Hras or Nras, promotes the expansion of an endodermal stem/progenitor cell and blocks its differentiation. Thus, Kras may uniquely contribute to the initiation of tumors in endodermally-derived tissues by expanding a stem/progenitor cell population. PMID- 18418067 TI - p14ARF is capable of promoting HIV-1 tat degradation. AB - The p14(ARF) tumor suppressor functions as 'oncogenic checkpoint' that prevents unrestricted cellular proliferation in response to oncogenic signaling. Albeit, the major pathway through which ARF operates is the ARF-Mdm2-p53 axis, ARF directly binds to and inactivates transcription function of a number of DNA-bound activators. In the present study we show that p14(ARF) inhibits transcription activation of HIV-1 LTR promoter activity by Tat protein. Tat protein is a RNA bound transcriptional activator whose function is strictly required for HIV-1 replication. We determined that p14(ARF) inhibits Tat transactivation of HIV-1 LTR by promoting Tat degradation via an ubiquitin-independent pathway. PMID- 18418068 TI - Live cell imaging of XLF and XRCC4 reveals a novel view of protein assembly in the non-homologous end-joining pathway. AB - XLF, also known as Cernunnos, is a newly identified core factor of the non homologous end-joining (NHEJ) pathway for DNA double-strand breaks (DSBs) repair. XLF is known to stimulate DNA ligase IV in vitro through its interaction with XRCC4. Here, we outline the key findings on the dynamic behavior of XLF and XRCC4 at DSBs in living cells. XLF is quickly recruited to DSBs in the absence of XRCC4 or DNA-PKcs. The recruited XLF molecules constantly exchange at DSBs, and XRCC4 modulates the exchange rate of the recruited XLF. XRCC4 can be recruited to DSBs without DNA-PKcs, but DNA-PKcs stabilizes the recruited XRCC4. These observations are inconsistent with the prevailing concept that NHEJ proteins are sequentially recruited to DSBs, which is mainly supported by in vitro evidence. We propose a novel two-phase model for the assembly of NHEJ factors to DSBs in vivo. XLF, XRCC4 and DNA-PKcs are independently recruited to Ku-bound DSBs. The recruited factors are assembled into a large complex, in which the protein interactions observed in vitro define the stability of the recruited factors. This new view has broad implications for the mechanism of DSB sensing and functional protein assembly in the NHEJ pathway. PMID- 18418069 TI - Analysis of DBC1 and its homologs suggests a potential mechanism for regulation of sirtuin domain deacetylases by NAD metabolites. AB - Deleted in Breast Cancer-1 (DBC1) and its paralog CARP-1 are large multi-domain proteins, with a nuclear or perinuclear localization, and a role in promoting apoptosis upon processing by caspases. Recent studies on human DBC1 show that it is a specific inhibitor of the sirtuin-type deacetylase, Sirt1, which deacetylates histones and p53. Using sensitive sequence profile searches and HMM HMM comparisons we show that the central conserved globular domain present in the DBC1 and it homologs from diverse eukaryotes is a catalytically inactive version of the Nudix hydrolase (MutT) domain. Given that Nudix domains are known to bind nucleoside diphosphate sugars and NAD, we predict that this domain in DBC1 and its homologs binds NAD metabolites such as ADP-ribose. Hence, we propose that DBC1 and its homologs are likely to regulate the activity of SIRT1 or related deacetylases by sensing the soluble products or substrates of the NAD-dependent deacetylation reaction. The complex domain architectures of the members of the DBC1 family, which include fusions to the RNA-binding S1-like domain, the DNA binding SAP domain and EF-hand domains, suggest that they are likely to function as integrators of distinct regulatory signals including chromatin protein modification, soluble compounds in NAD metabolism, apoptotic stimuli and RNA recognition. PMID- 18418071 TI - Inhale to survive, cycle or die? Carbon monoxide and cellular proliferation. AB - Heme oxygenase-1 (HO-1) and carbon monoxide (CO) modulate inflammation, proliferation/cell cycle and survival in a host of pathophysiological situations by reestablishing homeostasis. While several target genes and signaling pathways have now been elucidated that participate in HO-1/CO mediated protection, the events that occur in response to HO-1/CO under cellular stressors remain poorly understood particularly as they relate to therapeutic effects. Clearly there are differences among cell and tissue types driven by variations in basal gene expression profiles and more importantly under different activation states. In these instances where HO-1/CO mediate cytoprotection and restore homeostasis, critical regulatory and signaling mechanisms are in place to efficiently direct the cellular response. We propose that the HO-1 system acts as a biosensor for the cell. A fascinating aspect of the pleiotropic effects of the HO-1 system and the metabolic products involves the concept that its functional response befits the circumstances in which it finds itself whether prophylactically or therapeutically so as to ensure continued survival. This aspect of HO-1 and specifically the cellular response to CO as it relates to cell cycle and proliferation will be discussed in detail in this perspective. PMID- 18418070 TI - Modulation of 14-3-3 interaction with phosphorylated histone H3 by combinatorial modification patterns. AB - Post-translational modifications of histones are determining factors in the global and local regulation of genome activity. Phosphorylation of histone H3 is globally associated with mitotic chromatin compaction but occurs in a much more restricted manner during interphase transcriptional regulation of a limited subset of genes. In the course of gene regulation, serine 10 phosphorylation at histone H3 is targeted to a very small fraction of nucleosomes that is highly susceptible to additional acetylation events. Recently, we and others have identified 14-3-3 as a binding protein that recognizes both phosphorylated serine 10 and phosphorylated serine 28 on histone H3. In vitro, the affinity of 14-3-3 for phosphoserine 10 is weak but becomes significantly increased by additional acetylation of either lysine 9 or lysine 14 on the same histone tail. In contrast, the histone H3S28 site matches elements of 14-3-3 high affinity consensus motifs. This region mediates an initial stronger interaction that is less susceptible to modulation by "auxiliary" modifications. Here we discuss the binding of 14-3-3 proteins to histone H3 in detail and putative biological implications of these interactions. PMID- 18418072 TI - SET8 plays a role in controlling G1/S transition by blocking lysine acetylation in histone through binding to H4 N-terminal tail. AB - We report evidence suggesting that methyltransferase SET8 plays a novel role in regulating cell cycle by suppressing DNA replication through histone binding. First, the distribution of SET8 is strongly cell cycle-dependent. SET8 was concentrated in the nucleus during G(1) and G(2) phases, and was excluded from the nucleus during S phase. Second, at G(1)/S transition, SET8 was degraded through ubiquitination via SCF/Skp2. Third, it was evident that the SET8 binds to the H4 N-terminal tail (H4NT) and blocks the acetylation of lysine residues K5, K8 and K12 of histone H4 during G(1). Such a blockage can hinder DNA replication. Fourth, SET8 binds to hypoacetylated but not hyperacetylated H4NT. Finally, overexpressing the histone-binding domain of SET8 appeared to suppress DNA replication and arrest the cell cycle before the G(1)/S transition. Taken together, these findings suggest that SET8 can be a negative regulator of DNA replication and the destruction of SET8 is required for the onset of S phase. PMID- 18418073 TI - TIPT, a male germ cell-specific partner of TRF2, is chromatin-associated and interacts with HP1. AB - The differentiation process of spermatogenesis is based on a finely timed program of transcriptional regulation and chromatin remodeling. Male germ cells utilize specialized transcription complexes, which display specific differences in the components of the general transcription machinery. The TATA-binding protein (TBP) related protein 2 (TRF2) is essential for progression in spermiogenesis and for the structuring of the chromocenter, an heterochromatic structure unique of round spermatids. To decipher the molecular pathways of TRF2 action, we have searched for TRF2 partners in male germ cells. We have isolated TIPT (TRF2 interacting protein in testis), a relatively small protein that associates with TRF2 with affinity comparable to TFIIA. TIPT is uniquely expressed in testis, with a developmental pattern that temporally parallels the one of TRF2. Importantly, TIPT interacts also with HP1 proteins, thereby establishing an intriguing link between transcription and chromatin condensation. Association of TIPT with either TRF2 or HP1 occurs through the C-terminal domain in a mutually exclusive manner. These findings indicate that TIPT could contribute to the precise timing of the molecular events in male germ cells, specifically by linking transcription to chromatin remodeling in round spermatids. PMID- 18418074 TI - How do real tumors become resistant to cisplatin? AB - Lab research on cultured tumor cells selected for resistance to platinum compounds has turned up a diverse array of resistance mechanisms. In contrast, we recently found that mouse mammary tumors containing irrepairable null alleles of the Brca1 gene do not become resistant to cisplatin ever, although they invariably become resistant to a variety of other anti-cancer drugs. Each new treatment with cisplatin shrinks the tumor to a very small remnant, but relapse always occurs. The BRCA1 missing in these mouse tumors is essential for the homology-directed DNA repair (HR) that allows error-free repair of the duplex breaks caused by the excision of platin-DNA adducts. The mouse tumor results therefore raise the question whether the cisplatin resistance mechanisms identified in vitro can actually overcome an irreversible defect in DNA repair in real tumors. This question is underlined by recent analyses of tumor samples of patients with ovarian cancer that have uncovered a new platin resistance mechanism: these tumors were initially sensitive to platin through a defect in the BRCA2 gene, also required for HR, like BRCA1. Resistance in these patients, after an initial response of the tumor,-was due to secondary mutations in the defective BRCA2 gene, restoring BRCA2 function.(1,2) These clinical observations show the overriding importance of a functional HR system for tumor cells to survive platin-induced DNA lesions. Taken together with the mouse mammary tumor data, these observations raise the possibility that proliferating cells have no readily available mechanism to escape from cisplatin DNA damage once their HR is irreversibly inactivated. PMID- 18418075 TI - DNA replication in early S phase pauses near newly activated origins. AB - During the S phase of the cell cycle, the entire genome is replicated. There is a high level of orderliness to this process through the temporally and topologically coordinated activation of many replication origins situated along chromosomes. We investigated the program of replication from origins initiating in early S phase by labeling synchronized normal human fibroblasts (NHF1) with nucleotide analogs for various pulse times and measuring labeled tracks in combed DNA fibers. Our analysis showed that replication forks progress 9-35 kilobases from newly initiated origins, followed by a pause in synthesis before replication resumes. Pausing was not observed near origins that initiated in the middle of S phase. No evidence for pausing near origins was found at the beginning of the S phase in glioblastoma T98G cells. Treatment with the S phase checkpoint inhibitor caffeine abrogated pausing in NHF1 cells in early S phase. This suggests that pausing may comprise a novel aspect of the intra-S phase checkpoint pathway or a related new early S checkpoint. Further, it is possible that the loss of this regulatory process in cancer cells such as T98G could be a contributing factor in the genetic instability that typifies cancers. PMID- 18418076 TI - Targeting Plk1 to chromosome arms and regulating chromosome compaction by the PICH ATPase. AB - During mitosis, chromosomes undergo dynamic structural changes that include condensation of chromosomes-the formation of individual compact chromosomes necessary for faithful segregation of sister chromatids in anaphase. Polo-like kinase 1 (Plk1) regulates multiple mitotic events by binding to targeting factors at different mitotic structures in a phosphorylation dependent manner. In this study, we report the identification of a putative ATPase that targets Plk1 to chromosome arms during mitosis. PICH (Plk1-interacting checkpoint "helicase") displays a temporal localization on chromosome arms and kinetochores during early mitosis. Interaction with PICH recruits Plk1 to chromosome arms and disruption of this interaction abolishes Plk1 localization on chromosome arms. Moreover, depletion of PICH or overexpression of PICH mutant that is defective in Plk1 binding or ATP binding causes defects in mitotic chromosome compaction, formation of anaphase bridge and cytokinesis failure. We provide data to show that both PICH phosphorylation and its ATPase activity are required for mitotic chromosome compaction. Our study provides a mechanism for targeting Plk1 to chromosome arms and suggests that the PICH ATPase activity is important for the regulation of mitotic chromosome architecture. PMID- 18418077 TI - CDK1 inhibitors antagonize the immediate apoptosis triggered by spindle disruption but promote apoptosis following the subsequent rereplication and abnormal mitosis. AB - Spindle-disrupting agents and CDK inhibitors are important cancer therapeutic agents. Spindle toxins activate the spindle-assembly checkpoint and lead to sustained activation of CDK1. Different published results indicate that CDK1 activity is either important or dispensable for the cytotoxicity associated with spindle disruption. Using live cell imaging and various approaches that uncoupled mitotic events, we show that apoptosis was induced by both prolonged nocodazole treatment as well as by inhibition of CDK1 activity after a transient nocodazole block. However, distinct mechanisms are involved in the two types of cell death. The massive apoptosis triggered by nocodazole treatment requires the continuous activation of cyclin B1-CDK1 and is antagonized by premature mitotic slippage. By contrast, apoptosis induced by nocodazole followed by CDK inhibitors occurred after rereplication and multipolar mitosis of the subsequent cell cycle. The presence of dual mechanisms of cytotoxicity mediated by spindle disruption and CDK inhibition may reconcile the various apparent inconsistent published results. These data underscore the essential role of cyclin B1-CDK1 as the basis of apoptosis during mitotic arrest, and the role of mitotic slippage and abnormal mitosis for apoptosis at later stages. PMID- 18418078 TI - Oxidative stress induces cell cycle-dependent Mre11 recruitment, ATM and Chk2 activation and histone H2AX phosphorylation. AB - DNA damage response recruits complex molecular machinery involved in DNA repair, arrest of cell cycle progression, and potentially in activation of apoptotic pathway. Among the first responders is the Mre11- (MRN) complex of proteins (Mre11, Rad50, Nbs1), essential for activation of ATM; the latter activates checkpoint kinase 2 (Chk2) and phosphorylates histone H2AX. In the present study the recruitment of Mre11 and phosphorylation of ATM, Chk2 and H2AX (gammaH2AX) detected immunocytochemically were measured by laser scanning cytometry to assess kinetics of these events in A549 cells treated with H(2)O(2). Recruitment of Mre11 was rapid, peaked at 10 min of exposure to the oxidant, and was of similar extent in all phases of the cell cycle. ATM and Chk2 activation as well as H2AX phosphorylation reached maximum levels after 30 min of treatment with H(2)O(2); the extent of phosphorylation of each was most prominent in S-, less in G(1)-, and the least in G(2)M- phase cells. A strong correlation between activation of ATM and Chk2, measured in the same cells, was seen in all phases of the cycle. In untreated cells activated Chk2 and Mre11 were distinctly present in centrosomes while in interphase cells they had characteristic punctate nuclear localization. The punctate expression of activated Chk2 both in untreated and H(2)O(2) treated cells was accentuated when measured as maximal pixel rather than integrated value of immunofluorescence (IF) per nucleus, and was most pronounced in G(1) cells, likely reflecting the function of Chk2 in activating Cdc25A. Subpopulations of G(1) and G(2)M cells with strong maximal pixel of Chk2-Thr68(P) IF in association with centrosomes were present in untreated cultures. Cytometric multiparameter assessment of the DNA damage response utilizing quantitative image analysis that allows one to measure inhomogeneity of fluorochrome distribution (e.g., maximal pixel) offers unique advantage in studies of the response of different cell constituents in relation to cell cycle position. PMID- 18418079 TI - The CK2 phosphorylation of catalytic domain of Cdc34 modulates its activity at the G1 to S transition in Saccharomyces cerevisiae. AB - The ubiquitin-conjugating enzyme Cdc34 was recently shown to be phosphorylated by CK2 on the C-terminal tail. Here we present novel findings indicating that in budding yeast CK2 phosphorylates Cdc34 within the N-terminal catalytic domain. Specifically, we show, by direct mass spectrometry analysis, that Cdc34 is phosphorylated in vitro and in vivo by CK2 on Ser130 and Ser167, and that the phosphoserines 130 and 167 are not present after CK2 inactivation in a cka1Deltacka2-8(ts) strain. CK2 phosphorylation of Ser130 and Ser167 strongly stimulates Cdc34 ubiquitin charging in vitro. The Cdc34(S130AS167A) mutant shows a basal ubiquitin charging activity which is indistinguishable from that of wild type but is not activated by CK2 phosphorylation and its expression fails to complement a cdc34-2(ts) yeast strain, supporting a model in which activation of Cdc34 involves CK2-mediated phosphorylation of its catalytic domain. PMID- 18418080 TI - PDGF/VEGF system activation and angiogenesis following initial post ovariectomy meningeal microvessel loss. AB - Recently we demonstrated that cessation of ovarian hormone production causes dramatic vascular remodeling in meningeal microvascular networks characterized by a significant microvessel loss. Further, even two months post ovariectomy (OVX), dura mater microvessels remain destabilized due to a decline in estrogen-mediated angiopoietin-1 (Ang-1) expression and Ang-1/Tie-2 signaling. Such destabilized microvessels could be susceptible to either further regression or angiogenesis. In this study, we tested the hypothesis that initial estrogen-dependent loss of meningeal microvessels following OVX triggers stromal and vascular hypoxic responses aiming at restoring dura microvasculature. We demonstrate that two months post OVX, there is an activation of the hypoxia-inducible factor-1alpha (HIF-1alpha) and PDGF/VEGF system in the dura mater stroma and microvasculature of experimental animals accompanied by a shift in the balance between PI3K and PLCgamma activity downstream of PDGF/VEGF signaling toward PI3K. It appears that the latter serves as a molecular switch favoring angiogenic responses rather than further regression of destabilized microvessels. Indeed, consistent with this idea, we have found a considerable angiogenic activity in meningeal microvascular networks that previously underwent regression. These results indicate that angioadaptation of meningeal microvessels in response to cessation of ovarian hormone production is not a unidirectional, but a very complex multi-stage process regulated on many levels. The implication of this study is that therapeutic interventions, including estrogen-based hormone replacement therapy, with physiological angioadaptation in postmenopausal or post OVX women need to be approached with the extreme caution. PMID- 18418081 TI - Mechanism of relapse in pediatric acute lymphoblastic leukemia. AB - Relapse following initial chemotherapy remains a barrier to survival in approximately 20% of children suffering from acute lymphoblastic leukemia (ALL). Recently, to investigate the mechanism of relapse, we analysed clonal populations in 27 pairs of matched diagnosis and relapse ALL samples using PCR-based detection of multiple antigen receptor gene rearrangements. These clonal markers revealed the emergence of apparently new populations at relapse in 13 patients. In those cases where the new 'relapse clone' could be detected in the diagnosis population, there was a close correlation between length of first remission and quantity of the relapse clone in the diagnosis sample. A shorter length of time to first relapse correlated with a higher quantity of the relapsing clone at diagnosis. This observation, together with demonstrated differential chemosensitivity between sub-clones at diagnosis, indicates that relapse in ALL patients may commonly involve selection of a minor intrinsically resistant sub clone that is undetectable by routine PCR-based methods. From a clinical perspective, relapse prediction may be improved with strategies to detect minor potentially resistant sub-clones early during treatment, hence allowing intensification of therapy. Together with the availability of relevant in vivo experimental models and powerful technology for detailed analysis of patient specimens, this new information will help shape future experimentation towards targeted therapy for high-risk ALL. PMID- 18418082 TI - HOPS is an essential constituent of centrosome assembly. AB - Centrosomes direct microtubule organization during cell division. Aberrant number of centrosomes results from alteration of its components and leads to abnormal mitoses and chromosome instability. HOPS is a newly discovered protein isolated during liver regeneration, implicated in cell proliferation. Here, we provide evidence that HOPS is an integral constituent of centrosomes. HOPS is associated with classical markers of centrosomes and found in cytosolic complexes containing CRM-1, gamma-tubulin, eEF-1A and HSP70. These features suggest that HOPS is involved in centrosome assembly and maintenance. HOPS depletion generates supernumerary centrosomes, multinucleated cells and multipolar spindle formation leading to activation of p53 checkpoint and cell cycle arrest. The presence of HOPS in cytosolic complexes supports that centrosome proteins might be preassembled in the cytoplasm to then be rapidly recruited for centrosome duplication. Altogether these data show HOPS implication in the control of cell division. HOPS contribution appears relevant to understand genomic instability and centrosome amplification in cancer. PMID- 18418083 TI - ZM 447439 inhibition of aurora kinase induces Hep2 cancer cell apoptosis in three dimensional culture. AB - Mitotic Aurora kinases are essential for accurate chromosome segregation during cell division. Forced overexpression of Aurora kinase results in centrosome amplification and multipolar spindles, causing aneuploidy, a hallmark of cancer. ZM447439 (ZM), an Aurora selective ATP-competitive inhibitor, interferes with the spindle integrity checkpoint and chromosome segregation. Here, we showed that inhibition of Aurora kinase by ZM reduced histone H3 phosphorylation at Ser10 in Hep2 carcinoma cells. Multipolar spindles were induced in these ZM-treated G(2)/M arrested cells with accumulation of 4N/8N DNA, similar to cells with genetically suppressed Aurora-B. Cells subsequently underwent apoptosis, as assessed by cleavage of critical apoptotic associated protein PARP. Hep2 cells formed a tumor like cell mass in 3-dimensional matrix culture; inhibition of Aurora kinase by ZM either destructed the preformed cell mass or prevented its formation, by inducing apoptotic cell death as stained for cleaved caspase-3. Lastly, ZM inhibition of Aurora kinase was potently in association with decrease of Akt phosphorylation at Ser473 and its substrates GSK3alpha/beta phosphorylation at Ser21 and Ser9. Together, we demonstrated that Aurora kinase served as a potential molecular target of ZM for more selective therapeutic cancer treatment. PMID- 18418084 TI - Intronic CpG content and alternative splicing in human genes containing a single cassette exon. AB - Clinical data provide evidence for the association of missplicing with methyl binding protein mutations and inhibition of methylation. In this study, we analyzed a 373 human gene database containing a single alternatively spliced exon (cassette) and 1,039 constitutive introns, and showed that CpG frequencies are higher in alternative compared to constitutive introns, particularly in donors preceding cassette exons (p < 0.0001). Donors with more than three CpGs within 50 nt from splice junctions are mostly upstream alternative (21.83% vs. 3.21% for constitutive and 4.68% for downstream introns). Significant differences are also observed beyond the 7(th) nucleotide of the donors. Upstream CpG-rich motifs are not related to Alus, while the latter are frequent in downstream donors. The association of epigenetic modification sites and alternative splicing, indicated above, is not reflected in the computationally obtained splicing potentials. PMID- 18418085 TI - ZAP-mediated mRNA degradation. AB - The zinc-finger antiviral protein (ZAP) is a host factor that inhibits the replication of many viruses by preventing the accumulation of viral mRNAs in the cytoplasm. ZAP specifically binds to the viral mRNA and recruits the cellular RNA degradation machinery to degrade the target RNA. In this article, we will review the work to date in understanding the mechanisms by which ZAP promotes viral mRNA decay and discuss future research directions to further investigate the function and underlying mechanisms of ZAP. PMID- 18418086 TI - Identification of novel guide RNAs from the mitochondria of Trypanosoma brucei. AB - The majority of mitochondrial mRNAs in African trypanosomes are subject to an RNA editing reaction, which is characterized by the insertion and/or deletion of U nucleotides only. The reaction creates functional mRNAs and is catalyzed by a high molecular mass enzyme complex, the editosome. Editosomes interact with a unique class of small non-coding, 3'-oligouridylated (oU) RNAs, so-called guide RNAs (gRNAs). Guide RNAs function as transacting templates in the U deletion/insertion reaction and thus, represent key components in the reaction cycle. Furthermore, by utilizing different gRNAs, alternative editing events can take place, thereby expanding the protein diversity in the mitochondria of the parasites. In this study, we have analyzed small, non-coding mitochondrial transcripts from Trypanosoma brucei. By generating cDNA libraries from size selected RNA populations we identified 51 novel oU-RNAs. For 29 of these RNAs we were able to predict cognate mRNA targets. By Northern blot analysis, we verified the expression of 22 of these oU-RNAs and demonstrate that they share all known gRNA characteristics. Five of these 51 putative gRNAs are characterized by single mismatches to their cognate, fully edited mRNA sequences suggesting that they could act as gRNAs for alternative editing events. PMID- 18418087 TI - Roles for Srp72p in assembly, nuclear export and function of the signal recognition particle. AB - Co-translational protein targeting to the endoplasmic reticulum is catalysed by the signal recognition particle, a conserved ribonucleoprotein. Key activities of SRP--signal sequence binding, and inhibition of ribosomal translation elongation- require interactions of SRP with distant locations on the ribosome. A heterodimer of Srp72p and Srp68p localise to the central portion of the SRP complex, and may co-ordinate its activities. A series of mutations within Srp72p were examined individually, in combination and in the presence of mutations within SRP RNA. In this analysis mutations within Srp72p fell into two groups, identifying separate interactions/functions of the protein. Much of Srp72p is predicted to be alpha helical tetratricopeptide repeat motifs, with the C-terminus forming a separate unstructured region. Mutations towards the C-terminal end of the alpha helical region reveal a specific genetic interaction with a conserved motif in the central helix of SRP RNA. In contrast, mutations within the C-terminal region of Srp72p have genetic interactions across the RNA. Many mutant combinations impaired function rather than inhibiting assembly of SRP. However, one specific combination of Srp72p and SRP RNA mutations led to accumulation of pre-SRP in the nucleus. We conclude that Srp72p has at least two functions that are individually redundant and that the conformation of the complex is critical for efficient completion of its biogenesis. PMID- 18418088 TI - Perivascular epithelioid cell tumor (PEComa) of the uterus: an outcome-based clinicopathologic analysis of 41 reported cases. AB - The uterus and retroperitoneum have emerged as the most frequently reported anatomic sites of origin of perivascular epithelioid cell tumors (PEComas), a poorly defined neoplasm that is characterized by varying amounts of spindle and epithelioid cells with clear to eosinophilic cytoplasm that display immunoreactivity for melanocytic markers, most frequently HMB-45. Published reports on 41 previously reported uterine PEComas are reviewed in this report. Of these 41 cases, 31 originating in the corpus and for which there was adequate follow-up information (or clinical malignancy) were categorized into 2 groups: (1) a malignant group that was comprised of cases associated with patient death of disease and/or clinical malignancy as evidenced by local and/or distant extension outside of the uterus (n=13, group 1) and (2) a "nonmalignant" group of cases in which neither of the above features were present (n=18, group 2). Groups 1 and 2 did not significantly differ regarding duration of follow-up (25 mo vs. 24.3 mo, respectively, P=0.9) or patient age (45.61 y vs. 43.46 y, respectively, P=0.7). Five of the group 1 patients experienced distant (extra-abdominal) metastases. The group 1 tumors were significantly larger than the group 2 tumors (averages 9.6 cm vs. 4.67 cm respectively, P=0.04); however, there were no size thresholds that, in of themselves, reliably classified 75% or more of the cases in both groups. Coagulative necrosis was highly associated with group 1, being present in 82% of cases as compared with only 11.8% of group 2 cases (P=0.0002). Eighty-eight percent of the group 2 cases had a mitotic rate of 1/10 HPF and/or coagulative necrosis are features that, if present, raise the definite potential for aggressive behavior. PMID- 18418089 TI - Dermatopathology for the surgical pathologist: a pattern based approach to the diagnosis of inflammatory skin disorders (part I). AB - The accurate diagnosis of inflammatory conditions in dermatopathology requires integrating the histopathologic findings with the clinical features. This can be difficult when skin biopsies are seen infrequently and the terminology is unfamiliar, as in a general surgical pathology practice. This 2-part review of inflammatory cutaneous conditions presents a pattern-based approach to the diagnosis of dermatologic disorders and reviews the salient clinical and histologic features. Part I will focus on the spongiotic, psoriasiform, interface, and bullous patterns, whereas Part II will review the perivascular, vasculitis, panniculitis, and nodular and diffuse patterns. PMID- 18418090 TI - Expression of helix-loop-helix proteins in classical hodgkin lymphoma: a possible explanation for a characteristic immunophenotype. PMID- 18418091 TI - An attractive therapeutic target, mTOR pathway, in ALK+ anaplastic large cell lymphoma. PMID- 18418092 TI - Familial complete androgen insensitivity syndrome (Morris syndrome or testicular feminization syndrome) in 2 sisters. PMID- 18418098 TI - Review and update on inotropes and vasopressors. PMID- 18418100 TI - Breaking down the wall of silence to create healthy work environments: an interview with author Rosemary Gibson. PMID- 18418101 TI - What's in your toolbox? Considerations when selecting and evaluating instruments in clinical research. PMID- 18418102 TI - Geropharmacology: a primer for advanced practice acute care and critical care nurses, part I. AB - Advanced practice nurses' challenge in managing older adults' medication regimens from an evidence base is difficult because older adults are vulnerable to medication errors and adverse drug reactions related to a number of factors. Predicting patients' responses to drugs is compounded during critical illness, adding to the heterogeneity and unpredictability of drug effects that are prevalent premorbidly. In the first part of this 2-part continuing education series, sources of medication errors and older adults' vulnerability are discussed, including normal changes of aging affecting pharmacokinetics and pharmacodynamics, polypharmacy, self-medicating, patient-family noncompliance, and inappropriately prescribed medications. In the second part, drug classes and drugs posing particular problems for older adults and cautions for acute care and critical care nurses who manage the medications of older adults are highlighted. PMID- 18418104 TI - Pediatric obesity in acute and critical care. AB - Pediatric obesity has reached epidemic proportions in the United States. Significant obesity-related comorbidities are being noted at earlier ages and often have implications for the acute and critically ill child. This article will review the latest in epidemiologic trends of pediatric obesity and examine how it affects multisystem body organs. The latest data evaluating the specific effects of obesity on acute and critically ill children will be reviewed. Available nonpharmacologic, pharmacologic, and surgical strategies to combat pediatric obesity will be discussed. PMID- 18418105 TI - Measurement of thoracic fluid content in heart failure: the role of impedance cardiography. AB - Current guidelines for assessing the fluid status of patients with heart failure include subjective physical findings, which often occur late in decompensation, and objective pulmonary artery catheter measurements, whose use is controversial in patients with heart failure. Impedance cardiography, which assesses cardiac function by measuring the opposition to an alternating electric current in the thorax, indicates the amount of fluid in the thorax. It is used externally by employing electrodes on the thorax but can also be used internally by electrodes within a cardiac pacemaker. Recent studies have suggested that one of its direct measurements, thoracic impedance, may identify patients at risk for decompensation of heart failure, but because these measurements inconsistently correlate with pulmonary artery catheter measurements and study sample sizes are small, further research is required before its use can be recommended. PMID- 18418106 TI - An online dual-certification program in advanced practice nursing and nursing education: one university's response to faculty shortages. AB - Educating nurses for the healthcare delivery workforce is stymied as qualified applicants to nursing programs are being turned away. Although applications to baccalaureate programs have increased, between 41,683 and 147,000 undergraduate and graduate applicants were turned away from nursing education programs in 2005 due largely to shortages of nursing faculty. In this article, the evidence-based rationale for the development of a dual-certification program for the preparation of clinical nurse specialists and nurse educators is described. Because faculty shortages are nationwide, we developed the program to be delivered, in its entirety, online. Standardized data collection methods for evaluating student progress and their achievement of competencies expected of clinical nurse specialists and nurse educators are provided. The program may be a model for preparing clinically competent nurse educators who prefer practice settings to full-time faculty positions. PMID- 18418107 TI - The shortage of nurses and nursing faculty: what critical care nurses can do. AB - Nurses are needed more than ever to support the healthcare needs of every American. Nurses make up the greatest single component of hospital staff. In 2004, of the almost 3 million nurses in the United States, 83% were employed in nursing, and 58% of those were employed full-time. However, a severe shortage of nurses exists nationwide, putting the safe, effective healthcare of Americans in jeopardy. The concurrent shortage of nursing faculty has significant impact on the potential for admitting and graduating sufficient numbers of nursing students to address the shortage of prepared nurses. A close examination of the demographics of the 3 million nurses provides a context for an in-depth discussion of strategies that critical care nurses can employ to help alleviate the nursing and nurse faculty shortages. PMID- 18418108 TI - A faculty practice plan for the acute care and critical care nurse. AB - This article discusses faculty clinical practice and explains the steps that one group of nursing faculty used to develop a faculty practice plan at a private college of nursing in a small, faith-based university in a midwestern city. The goal of the faculty practice plan was to link faculty interests and advanced practice skills with targeted populations and community needs. This was done to provide healthcare services to schools, agencies, health and human services programs, and populations in need. Examples of faculty roles in the faculty practice domains of educator, researcher, and expert clinician in an acute care setting illustrate how nurses can individualize their participation in the faculty practice plan to support their professional career goals. Faculty practice plans can benefit not only nursing faculty but also the college of nursing, its students, and its community and populations of interest. PMID- 18418109 TI - Revitalizing academic-service partnerships to resolve nursing faculty shortages. AB - Vacancy rates of nursing faculty are on the increase as fewer educators are being prepared and nurses at the bedside are in short supply. Advanced practice nurses- uniquely suited to provide clinical education for undergraduate and graduate students--are underutilized. However, without release time, preparation as educators, ongoing support from education and administration, and remuneration for providing both evidence-based education and evidence-based patient care, they can hardly be expected to assume additional responsibilities. This article reviews academic-service partnerships, noting a general lack of published efficacy and cost-effectiveness data, and highlights 21st-century innovations for academia and service to share resources for the benefit of all stakeholders- especially patients and students. PMID- 18418110 TI - ECG diagnosis of acute coronary syndrome. PMID- 18418111 TI - Notes from the board: President's message. PMID- 18418112 TI - Clinical nurse specialist as attending nurse? PMID- 18418113 TI - Ethical regulations in research. PMID- 18418114 TI - Chlorhexidine wipes: the new weapon against surgical site infections? PMID- 18418115 TI - Influencers among us: a practical approach for leading change. PMID- 18418116 TI - Implementing the clinical nurse specialist role in a regional state psychiatric hospital. AB - Clinical nurse specialists (CNSs) are essential to enhancing psychiatric-mental health nursing within state psychiatric hospital settings. This article presents a project focused on the CNSs in a state psychiatric hospital to expand their clinical role to increase the quality of care to individuals with serious mental illness and their numbers in other state psychiatric hospitals. As the patient population served in these settings becomes more complex, it is critical that advanced practice nurses are provided with appropriate updated state-of-the-art advanced knowledge and skills to collaborate within the interdisciplinary team. Furthermore, because of the complexity of the client and systems characteristics, the CNSs' clinical leadership is critical to promote the best practices in direct care services as well as provide support to various levels of nursing through staff development and training, role modeling, and mentoring of new staff. PMID- 18418117 TI - Reducing preventable medication safety events by recognizing renal risk. AB - Evidence suggests that medication safety may be improved through more accurate assessment of renal function and appropriate dosing of renally cleared medications. The purpose of this article is to describe patient renal risk groups, associated medication errors, and ways that nurses can improve renal assessment. METHODS: Medication safety data were collected through voluntary reporting, computerized triggers, pharmacist surveillance, and retrospective chart review. Data were analyzed across 3 renal risk groups. RESULTS: Findings indicated that regarding the detected medication errors, elderly women were more likely to have hidden renal risk and that prescribing errors involving a wrong dose occurred more often in patients with high and hidden renal risk. Antibiotic and diabetic medications were the primary drug categories involved in these medication errors. RECOMMENDATIONS: Results indicated that identification of patients with hidden renal risk can be improved by routinely assessing serum creatinine and estimated creatinine clearance levels during renal assessments. Clinical nurse specialists can use this evidence to promote safer nursing care of renal patients. PMID- 18418119 TI - Safety work: initiating, maintaining, and terminating restraints. AB - PURPOSE: This study examines the variations and complexities in nurses' decision making about the initiation, maintenance, and termination of physical restraints. METHODS: This qualitative research used a semistructured interview of registered nurses on medical-surgical units at a midsized hospital in the Midwest. RESULTS: All the nurse respondents easily recalled caring for a patient in restraints and detailed in their accounts a complex trajectory of care that centered on safety. However, most nurses reported that the trajectory of restraint use was started by other departments, shifts, or nurses. The findings revealed a forceful interplay of patient, nurse, family, healthcare worker, and organizational factors that influence nurses' decision making about restraint use. CONCLUSIONS: This article discusses how a deeper understanding of the intricacies of the decision-making processes related to restraints can help clinical nurse specialists tailor education, impact policy, and serve as role models to reduce the use of restraints in hospitals. PMID- 18418120 TI - Clinical nurse specialist profile: Sherri Atherton. Interview by Jo Ellen Rust. PMID- 18418121 TI - There is a bridge and the Trebus Project: artists create hope and understanding for those with dementia. PMID- 18418124 TI - Elevated gluconeogenesis and lack of suppression by insulin contribute to cystic fibrosis-related diabetes. AB - The incidence of diabetes is high in cystic fibrosis (CF) and is an important cause of morbidity and mortality. Understanding the pathophysiology is imperative. Studies have documented increased endogenous (mostly hepatic) glucose production (HGP) but have not distinguished the relative contribution of gluconeogenesis (GNG). The purpose of this study was to quantitate GNG, to determine its contribution to high HGP, and to measure insulin's suppression of GNG. We recruited 31 adult CF subjects (age, 26.2+/-7.9 years; 12 female subjects) and quantified GNG by measuring the incorporation of H into the second and fifth carbons of glucose. Hepatic glucose production was measured using [6,6 H2]glucose. Protein breakdown was measured using [1-C]leucine. Data were compared with that from 11 healthy volunteers (age, 27.5+/-7.0 years) who underwent both GNG and clamp studies. Thirteen CF subjects and all controls had a hyperinsulinemic euglycemic clamp during measures of GNG. Other measures included glucose tolerance and glucagon and cortisol levels. Rate of GNG was higher in CF subjects than controls and comprised a greater percentage of fasting HGP (GNG as percent of HGP: CF=68%; controls=44%; P=0.034). Suppression of GNG by insulin was significantly lower in CF than in controls and was lower in CF subjects with abnormal glucose tolerance than in those with normal glucose tolerance. Gluconeogenesis correlated with protein breakdown. These studies suggest that high HGP in CF is mostly from elevated rates of GNG and that resistance to insulin's suppression of GNG may contribute to abnormal glucose tolerance in CF. PMID- 18418125 TI - Ethnic difference in sex gap in high-density lipoprotein cholesterol between Asian Indians and Whites. AB - OBJECTIVE: To study whether low plasma high-density lipoprotein cholesterol (HDL C) reported in Asian Indians is common in both men and women when compared with whites and whether it is related to increased body mass index (BMI) and plasma triglyceride concentration. DESIGN: We evaluated the lipid profile and prevalence of low HDL-C (<40 mg/dL in men and <50 mg/dL in women) in the following cohorts of normoglycemic 1404 men and 1817 women: Asian Indians living in rural India; urban Chennai, India; and Dallas, TX; and whites living in Dallas, TX. RESULTS: After adjustment for age, BMI, and smoking, HDL-C was not significantly different in Asian Indian men compared with whites. However, Asian Indian women had lower HDL-C compared with white women, and rural Asian Indian women had the lowest HDL C even in the absence of high triglycerides. Lean Asian Indian women with BMI of less than 23 kg/m had higher frequency of low HDL-C compared with lean white women with BMI of less than 25 kg/m (72%, 56%, 48%, and 25% in rural, urban, and Dallas Asian Indian and white women, respectively) and lean men (52%, 42%, 28%, and 35% in rural, urban, and Dallas Asian Indian and white men, respectively). Sex differences in HDL-C was estimated as 6.6+/-0.5 mg/dL for Asian Indians and 15.3+/-1.1 mg/dL for whites (P<0.0001 for sex difference in the 2 ethnic groups). CONCLUSIONS: Increased prevalence of low HDL-C independently of obesity or hypertriglyceridemia is observed in women but not in men of Asian Indian origin. The sex gap in HDL-C is significantly smaller in Asian Indians compared with whites independent of geographical location. PMID- 18418126 TI - Detection of alpha1 integrin in urine of patients with immunoglobulin A nephropathy. AB - BACKGROUND: The alpha1beta1 integrin is a cell surface membrane heterodimer composed of noncovalently linked alpha1 and beta1 polypeptides that is up regulated on activated and proliferating mesangial cells. METHODS: A double sandwich enzyme-linked immunosorbent assay that detects alpha1 integrin in a specific and dose-dependent manner at concentrations greater than 150 ng/mL was used to evaluate whether intact alpha1 polypeptides are secreted in the urine samples of 29 patients with various kidney diseases and in those of 5 healthy individuals. RESULTS: alpha1 Integrin was detected in 8 of the 29 patients including 3 of 3 patients with biopsy-proven immunoglobulin A nephropathy and 3 of 3 clinically suspected but non-biopsy-proven immunoglobulin A nephropathy with evidence of active nephritis. No alpha1 integrins were found in samples of 5 healthy controls. CONCLUSIONS: alpha1 Integrin polypeptides can be detected in human urine, particularly in immunoglobulin A nephropathy. Further extensive studies are required to clarify the significance of secretion of alpha1 integrins in urine of patients with kidney disease. PMID- 18418128 TI - An evaluation of low back-pain-related content in Canadian newspaper media. AB - STUDY DESIGN: Review of newspaper articles. OBJECTIVE: To assess the content of newspaper articles in 2 provinces in Canada to determine if rest or avoidance of activity is being recommended for back pain. SUMMARY OF BACKGROUND DATA: Inaccurate back pain beliefs in the general public may arise due to messages in the mass media. One persisting belief in Canada is that rest or activity avoidance is needed until back pain resolves. METHODS: We searched newspapers in 2 Canadian provinces via an electronic database for articles discussing back pain. Two trained raters used an article review template to indicate whether the article's main recommendation was to stay active, rest, was neutral (indicating a balance between rest and activity), or did not provide advice on level of activity during an episode of back pain. RESULTS: One hundred 29 articles were identified. The primary advice provided related to level of activity during an episode of back pain was stay active in 24% of articles, whereas no articles primarily recommended rest or avoidance of activity. Sixteen percent of articles were rated as neutral, indicating the authors suggested a balance between rest and activity. CONCLUSIONS: Back-pain-related newspaper articles do not carry messages that advocate rest or avoidance of activity, but rather highlight the importance of staying active during an episode or participating in exercise. PMID- 18418129 TI - Vertical atlantoaxial index: a new craniovertebral radiographic index. AB - STUDY DESIGN: An index for measuring the vertical relationship of atlas and axis is described. OBJECTIVE: Deduction and application of vertical atlantoaxial index (VAAI) for quantifying the vertical atlantoaxial relationship of atlas and axis and classifying basilar invagination (BI) based on the VAAI. SUMMARY OF BACKGROUND DATA: A number of craniospinal parameters have been described to quantify the amount of BI in relation to herniation of odontoid into the cranium. Successful treatment of BI by distraction of atlantoaxial facet joint, placement of a spacer to maintain the distraction, and lateral mass fixation has been described recently. The treatment is based on the understanding that BI is in fact a disease of the atlantoaxial facet joints and is a result of progressive vertical instability. With this new concept and technique that involves preserving all bony elements and reduction of BI in the form of reducing the vertical atlantoaxial subluxation, we thought it was prudent to formulate an index to quantify the relation of atlas and axis in the sagittal plane. METHOD: Mid-sagittal computerized tomography (CT) scan films of 90 cases of BI treated by us between October 1999 and May 2005 with distraction and lateral mass plate and screw fixation were analyzed before and after surgery. The age of the patients ranged from 8 to 55 years and the male:female ratio was 2.5:1. Additionally, mid sagittal CT scan films of hundred normal subjects in the same age group were analyzed as a control group. The VAAI was measured in all cases. VAAI is an index that measures the vertical relationship of the atlas and axis. The images were compiled and copies of the compilation were made. Two observers independently performed the measurements and intraobserver as well as interobserver agreement was assessed using the intraclass corelation (ICC) ) test (SigmaStat). RESULTS: The preoperative mean and mode values of VAAI in this series of patients were 0.53 (0.20 to 0.67) and 0.61, respectively. The postoperative mean and mode values of VAAI are 0.78 (range, 0.60 to 0.89) and 0.80, respectively. The mean and mode values of VAAI in general population were 0.80 (range, 0.76 to 0.85) and 0.80, respectively. The results in preoperative patients with BI demonstrated excellent intraobserver (ICC=0.96 and 0.98) and interobserver corelation coefficient (ICC=0.96). The results in normal subjects and postoperative CT scans of operated patients with BI also showed excellent intraobserver (ICC=0.97 and 0.98) and interobserver corelation coefficient (ICC=0.97). CONCLUSIONS: VAAI can be an excellent measurement tool for the assessment of relationship of atlas and axis. Nonrheumatoid BI can be graded and classified depending on the value of VAAI. PMID- 18418130 TI - Intradiscal electrothermal therapy (IDET) for low back pain in worker's compensation patients: can it provide a potential answer? Long-term results. AB - OBJECTIVE: This prospective study was conducted to evaluate improvements in pain and disability in a series of 53 consecutive worker's compensation patients with low back pain (LBP) after treatment with the intradiscal electrothermal therapy (IDET) procedure. MATERIALS AND METHODS: All patients seen in the out-patient clinic of the Spine Institute of Louisiana for LBP of discogenic origin were screened for eligibility to receive IDET procedure. A total of 134 patients were treated using IDET for their discogenic LBP during the study period. Fifty-three patients presented to us via the worker's compensation claim program. The outcomes of these 53 patients were analyzed statistically for the current study by physical examination and self-assessment questionnaires of pain and disability at baseline and at 12-months postprocedure. Pain and disability outcomes were assessed by visual analog scale (VAS) pain score and Oswestry disability index, respectively. RESULTS: The mean patient age was 41.83 years (range 20 to 61 y). Whites (52.8%), African-Americans (30.2%), and Hispanics (17%) formed the majority of population. Forty-nine percent were using narcotics. The first definitive end point was considered at 12 months after the procedure. Median follow-up period was 56 months (range 29 to 72 mo). A mean reduction (P<0.001) of 62.6% in the VAS score and 69.3% in the Oswestry scores was noted after IDET. The patient's initial VAS and Oswestry scores (P<0.05) significantly affected the final outcomes. About 47.2% of the patients had some degree of economic productivity and only 7 (initial 26) consumed narcotic analgesics. CONCLUSIONS: IDET procedure can be a useful, safe, and cost-effective option in the management of carefully selected workers compensation claimants with chronic LBP of discogenic etiology. PMID- 18418131 TI - Cervical kinematics after fusion and bryan disc arthroplasty. AB - INTRODUCTION: Disc arthroplasty has been shown to provide short-term clinical results that are comparable with those attained with traditional anterior cervical discectomy and fusion. One proposed benefit of arthroplasty is the ability to prevent or delay adjacent level operations by retaining motion at the target level and eliminating abnormal adjacent activity. This paper compares motion parameters for single-level anterior cervical discectomy and fusion and disc replacement patients at the index level and adjacent segments. METHODS: Radiographic data from patients enrolled in a prospective, randomized clinical trial were selected for kinematic assessment of cervical motion. All patients received either a single-level fusion with allograft and anterior cervical plate (Atlantis anterior cervical plate, n=13) or a single-level artificial cervical disc (Bryan Cervical Disc prosthesis, n=9) at either C5/C6 or C6/C7. Flexion, extension, and neutral lateral radiographs were obtained preoperatively, immediately postoperatively, and at regular intervals up to 24-month time points. Cervical vertebral bodies were tracked on the digital radiographs using quantitative motion analysis software (QMA, Medical Metrics) to calculate the functional spinal unit motion parameters including range of motion (ROM), translation, and center of rotation. If visible, the functional spinal unit parameters were obtained at the operative level, and also the level above and the level below. RESULTS: As expected, significantly (P<0.006 at 3, 6, 12, and 24 mo) more flexion/extension motion was retained in the disc replacement group than the plated group at the index level. The disc replacement group retained an average of 6.7 degrees at 24 months. In contrast, the average ROM in the fusion group was 2.0 degrees at the 3-month follow-up and gradually decreased to 0.6 degrees at 24 months. The flexion/extension ROM both above and below the operative level was not statistically different for the disc-replaced and fusion patients, however, mobility increased for both groups over time. The anterior/posterior translation that occurs with flexion/extension motion remained unchanged for the disc replacement group at the level above the target disc preoperatively and postoperatively. In contrast, the translation increased for the level above the fusion. At the 6-month follow-up, the increase in translation was significantly greater for patients that were fused (P<0.02) than for patients that received a disc replacement. This change was not significant at 12 months. DISCUSSION: Previous studies have shown the Bryan disc to maintain mobility at the level of the prosthesis. The long-term clinical benefit of maintenance of motion is postulated to be the ability to delay or avoid adjacent level operations. This study reveals that there is no difference in flexion/extension ROM at the level above and below either a fusion or Bryan arthroplasty. There is, however, an increase in anterior/posterior translation at the cephalad adjacent level in patients with arthrodesis while the Bryan arthroplasty retains normal translation for the same amount of flexion/extension at the adjacent level. CONCLUSION: The Bryan disc may delay adjacent level degeneration by preserving preoperative kinematics at adjacent levels. PMID- 18418132 TI - Raster stereography versus radiography in the long-term follow-up of idiopathic scoliosis. AB - STUDY DESIGN: Raster-stereographic and radiographic evaluation of idiopathic scoliosis without braces in a retrospective longitudinal long-term follow-up study. OBJECTIVE: To investigate the reliability and accuracy of raster stereography in comparison with radiography as the gold standard, using a longitudinal long-term study design in idiopathic scoliosis, to reduce the number of radiographs required during follow-up in scoliosis patients. SUMMARY OF BACKGROUND DATA: It has been confirmed that raster stereography produces reliable data in patients with conservatively and surgically treated idiopathic scoliosis, up to a Cobb angle of 80 degrees. This means that the method can be used to replace radiography during the follow-up in these patients. However, no data have yet been published on the use of raster stereography in a longitudinal setting during a long-term follow-up period in comparison with radiography as the gold standard. METHODS: Raster stereographs and digitized anterior-posterior radiographs of 16 patients with idiopathic scoliosis were studied retrospectively in a longitudinal study design, with a mean follow-up period of 8 years (range 3 to 10 y). Lateral vertebral deviation and vertebral rotation were measured between C7 and L4 using raster stereography and radiography, compared with Cobb angles, and correlated. RESULTS: During the follow-up period, the Cobb angle increased on average by 13 degrees. The progression of lateral vertebral deviation measured using both techniques, and that of vertebral rotation measured with radiography, was greater than that of the Cobb angle, whereas that of raster stereographic vertebral rotation was lower. However, there was an excellent correlation between the raster-stereographic and radiographic progression of these parameters (R2 >or=0.5). The mean difference between raster stereographs and radiographs was 3.21 mm for lateral vertebral deviation and 2.45 degrees for vertebral rotation. CONCLUSIONS: Using the parameters of lateral vertebral deviation and vertebral rotation, raster stereography accurately reflects the radiographically measured progression of idiopathic scoliosis during the long term follow-up, but these parameters are not directly comparable with the Cobb angle. In the follow-up of scoliosis patients, the authors would recommend a raster-stereographic examination every 3 to 6 months and a radiographic examination every 12 to 18 months only, provided that raster stereography does not show rapid deterioration of the scoliosis. The patient's radiation exposure can be reduced using this approach. PMID- 18418133 TI - Viscosupplementation in lumbar facet joint arthropathy: a pilot study. AB - BACKGROUND: Degeneration of the spine is a prevalent problem that generally advances with age, although its occurrence is not restricted to the elderly. Chronic low back pain is a common problem with several treatments, but rigorous evaluation of many interventions is still lacking. One of the most common methods of treating lumbar facet arthropathy is intra-articular injection of local anesthetic and steroid. However, in large joints, such as the knee, viscosupplementation has been shown to compare favorably to steroid in terms of symptom relief and duration of benefit. It is suggested that viscosupplementation may supersede steroid in treatment of symptomatic knee osteoarthritis. Hyaluronic acid (HA) therapy is still in its infancy in joints outside the knee, although some preliminary results are promising. OBJECTIVE: This was a pilot study to test the potential effectiveness of HA injection therapy in the treatment of lumbar facet joint arthritis. METHODS: Thirteen patients with symptomatic lumbar facet joint arthritis who met the inclusion criteria were prospectively recruited. Pretreatment evaluation of patients was by questionnaire, including the Visual Analog Score and Oswestry Disability Questionnaire. A single injection of HA into affected facet joints was then performed, with correct placement confirmed on fluoroscopy. The patients were similarly evaluated 6 weeks after treatment. RESULTS: Eighteen facets in 13 patients were injected with HA. At 6-week follow up, there was no significant improvement in pain when measured on the Visual Analog Score. There was also no significant improvement in the Oswestry Disability Questionnaire. CONCLUSIONS: Preliminary results from this pilot study do not demonstrate any benefit of viscosupplementation in the management of symptomatic lumbar facet arthropathy. PMID- 18418134 TI - Anterior lumbar interbody fusion for lumbosacral junction in steep sacral slope. AB - STUDY DESIGN: Retrospective study of surgical technique and clinical outcome. OBJECTIVES: To examine the technique and outcomes of anterior lumbar interbody fusion (ALIF) surgery for a lumbosacral junction in a steep sacral slope. SUMMARY OF BACKGROUND DATA: There are no studies on the outcome and technical pitfalls on ALIF surgery for a lumbosacral junction in a steep sacral slope. MATERIALS AND METHODS: Six female patients (mean age of 55.67 y; range, 42 to 69) who had a steep sacral slope underwent ALIF surgery for degenerative (2 patients) and spondylolytic (4 patients) spondylolisthesis. The average follow-up duration was 29.33 months (range, 27 to 33 mo). The following parameters were used to assess the outcomes: slip angle, slip percentage, sacral inclination, lumbosacral angle, sacral slope, and distance from the upper margin of the pubis symphysis to the sacral plane. The level of pain was measured using the visual analog pain scale score. The function was assessed using the Oswestry Disability Index (ODI) score. Satisfaction surveys were also carried out. Statistical analysis was performed using a Friedman test. A P value <0.05 was considered significant. RESULTS: The mean sacral inclination, lumbosacral angle, sacral slope, and distance from the upper margin of the pubis symphysis to the sacral plane were 37.34 degrees (range, 28.55 to 48.92 degrees), 12.20 degrees (range, 5.09 to 16.5 degrees), 40.70 degrees (range, 30.54 to 49.98 degrees), and 22.06 cm (range, 16.13 to 29.72 cm), respectively. The mean correction of slip percentage and slip angle was 35.46%, and 9.3 degrees, respectively. The mean visual analog pain scale score decreased from 8.5 (back pain) and 7.3 (leg pain) to 1.8 (back pain) and 1.8 (leg pain) after surgery (P=0.001). The mean ODI scores also reflected the improved status (ODI of 64.7 before surgery to 8.5 after surgery; P=0.001). The patient's satisfaction was relatively high. All the patients had radiographically solid fusion at the latest follow-up. There were no significant complications encountered in this study. CONCLUSIONS: In selected cases, a steep sacral slope may not be an absolute contraindication of ALIF. Moreover, the C-arm-guided reduction and cage insertion method is a reliable way of treating spondylolisthesis in those with a steep sacral slope. PMID- 18418135 TI - Biomechanical evaluation of paracoccygeal transsacral fixation. AB - STUDY DESIGN: This is a biomechanical study using human cadaveric lumbar spine. OBJECTIVE: To evaluate the biomechanics of paracoccygeal transsacral rod fixation. SUMMARY OF BACKGROUND DATA: Various types of transsacral fixation either by posterior and paracoccygeal approaches have been described in the literature. The biomechanical advantage of transsacral rod fixation is the preservation of supporting structures at L5-S1 level. No biomechanical data on human cadavers have been reported in the literature. The aim of this study is to evaluate the biomechanics of the transsacral rod fixation. METHODS: Six fresh human cadaveric L5-S1 motion segments (mean age 67.5 y; range 46 to 82 y ) were used in the study. Unconstrained and nondestructive pure moments in axial torsion (AT), lateral bending (LB), and flexion-extension (FE) were applied to each specimen after applying transsacral rod and after additional augmentation methods, including bilateral screws, facet screws, and pedicle screw and rod system. Range of motion (ROM) was calculated for each surgical treatment. The disc space was measured with lateral plain radiographs of intact specimens and after transsacral rod insertion to evaluate the amount of distraction. RESULTS: The mean ROM of the intact specimens was 3.5, 6.4, and 11.0 degrees in AT, LB, and FE, respectively. Standalone transsacral rod reduced ROM more than 40% compared with the intact condition (P=0.002). Bilateral screws further reduced the ROM in AT (64%) and LB (70%), but not in FE (53%). Both facet screws and pedicle screw and rod system achieved high construct stability under all loading conditions. The transsacral rod augmented with facet screws reduced ROM by 70%, 80%, and 90% compared with the intact condition. When augmented with pedicle screw and rod system, the transsacral rod reduced ROM by 73%, 87%, and 88% in AT, LB, and FE, respectively. There was no statistical difference between these 2 facet screws and pedicle screw and rod system (P>0.8). CONCLUSIONS: Transsacral rod fixation provides strong ligamentotaxis due to intact annulus. Standalone transsacral rod is able to reduce ROM significantly and achieve indirect decompression by distracting L5-S1 disc space. However, additional posterior fixation, such as facet screws or pedicle screws, is required to achieve better construct stability for successful fusion. PMID- 18418136 TI - Odontoid screw placement using Isocentric 3-dimensional C-arm fluoroscopy. AB - STUDY DESIGN: We describe the use of isocentric 3-dimensional fluoroscopy to place odontoid screws in 9 patients. OBJECTIVE: We wanted to show the benefits of using isocentric 3-dimensional fluroscopy in odontoid screw placement. SUMMARY OF BACKGROUND DATA: Odontoid screw fixation for treatment of type II odontoid fractures has gained popularity since its introduction in the early 1980s. During the last several years, a multitude of new techniques have improved the ease of odontoid screw placement, including biplanar fluoroscopy, cannulated screw systems, and beveled bedside-fixed retractor systems. The use of isocentric C-arm fluoroscopy can improve the ease and facilitate placement of odontoid screws. CLINICAL PRESENTATION: Nine patients, ranging in ages from 30 to 89 years, presented with type II odontoid fractures. All fractures were either nondisplaced or minimally displaced (<4 mm) and occurred as a result of acute trauma. No patient had evidence of transverse atlantal ligament disruption. METHODS: Isocentric 3-dimensional fluoroscopy, in conjunction with image-guided navigational software, was used to place 1 or 2 odontoid screws in each patient. Three-dimensional images were acquired intraoperatively, which were then reconstructed and uploaded to the navigational workstation. Screw trajectory was planned and performed with the use of tracked instruments. RESULTS: Successful screw placement, as judged by intraoperative computerized tomography, was attained in all 9 patients. CONCLUSIONS: Isocentric 3-dimensional fluoroscopy, in conjunction with an image-guided navigational software system, obviates the need for cumbersome biplanar fluoroscopy, allows for intraoperative image acquisition after surgical exposure, reduces intraoperative registration time, reduces both surgeon and patient radiation exposure, and allows immediate computerized tomographic imaging in the operating room to verify screw position. PMID- 18418137 TI - Validity of serum creatine kinase as a measure of muscle injury produced by lumbar surgery. AB - BACKGROUND AND PURPOSE: Serum creatine kinase (CK) concentrations have historically been used to investigate muscle disease and serious muscle damage, and there is a growing interest in the potential for a biochemical approach to quantifying skeletal muscle injury occurring in orthopedic surgeries and spinal injuries. The wide availability of CK measurement could foster spinal muscle injury research. However, measurement validity has never been systematically demonstrated in clinical settings. In this study, the validity of serum CK concentration elevation as an index of muscle injury was investigated using lumbar decompression surgery (LDS) as a model. SUBJECTS AND METHODS: Blood samples were obtained from 18 research volunteers drawn from the clinical population undergoing LDS. A baseline sample was taken in the preoperative waiting area. Each subject's highest CK concentration between 12 and 48 hours after surgery was used as the biochemical injury response. The surface area of muscle isolated (incision lengthxdepth) and strained by retraction was obtained for concurrent validity testing against biochemical measurement. RESULTS: The correlation between highest total CK concentration and muscle surface area was moderate (r=0.60) and significant (P<0.01). Correlations between surface area and CK at specific time points, revealed minimal loss of association at 12 hours (r=0.57) and 24 hours (r=0.58), but weaker correlations at 6 hours (r=0.45) and 48 hours (r=0.28) after injury. Analyses for proportions of each isoenzyme making up the total CK revealed that baseline and peak CK consisted almost exclusively of skeletal muscle CK (CK-MM), with minimal representation by heart muscle (CK MB), and brain (CK-BB). CONCLUSIONS: The findings provide support for the validity of serum CK measurement as an index of skeletal muscle injury caused by LDS, and demonstrate that LDS provides a useful model for measurement testing and development studies. PMID- 18418138 TI - Intradiscal electrothermal therapy (IDET) provides effective symptom relief in patients with discogenic low back pain. AB - STUDY DESIGN: Single-arm, prospective clinical trial. OBJECTIVE: To evaluate back pain severity, physical function, and quality of life outcomes in highly selected patients with lumbar discogenic pain treated with intradiscal electrothermal therapy (IDET). SUMMARY OF BACKGROUND DATA: Degeneration and disruption of the intervertebral disc can be the source of severe low back pain and the associated physical dysfunction. IDET is a minimally invasive treatment option in the continuum of care between conservative nonoperative management and spinal surgery. METHODS: Using magnetic resonance imaging, pressure-controlled discography, and postdiscogram computed tomography findings, 56 patients with lumbar discogenic pain were identified, underwent IDET treatment and followed for 20.5+/-4.4 months, on average. Outcomes included assessments of back pain severity by visual analog scale, sitting, standing, and walking tolerances, and health-related quality of life using the SF-36. RESULTS: Mean pain severity scores (visual analog scale) improved from 6.1+/-1.8 pretreatment to 2.4+/-2.6 at final follow-up (P=0.0001). Mean tolerance times (minutes) improved from 40.9+/ 40.6 to 84.5+/-54.4, 46.8+/-42.9 to 84.4+/-54.2, and 39.2+/-39.6 to 77.9+/-50.8 between baseline and final follow-up for sitting, standing, and walking, respectively (P=0.0001 for all comparisons). Seven of 8 quality of life domains showed significant (P=0.0001 for all comparisons) improvement over baseline. Forty-two patients (75%) were classified as a treatment success by virtue of a >or=2-point improvement in pain severity or a >or=10-point improvement in either the physical functioning or bodily pain domain of the SF-36. CONCLUSIONS: The findings of this study suggest that durable clinical improvements can be realized after IDET in highly selected patients with mild disc degeneration, confirmatory imaging evidence of annular disruption, and concordant pain provocation by low pressure discography. PMID- 18418139 TI - Pedicle screw placement with a free hand technique in thoracolumbar spine: is it safe? AB - STUDY DESIGN: Computerized tomography (CT) analysis of in vivo pedicle screw placement to determine their exact position in relation to the pedicle in thoracolumbar region (T10-L3). OBJECTIVE: To evaluate the clinical accuracy of the placement of thoracolumbar pedicle screws with a free hand technique after reviewing preoperative imaging. SUMMARY OF BACKGROUND DATA: CT scans have been used in research clinical settings to evaluate pedicle screw placement in thoracolumbar spine. MATERIAL AND METHODS: Ninety-eight consecutive patients who underwent posterior stabilization using 640 transpedicular screws by 1 surgeon from T10 to L3 were analyzed. The mean age was 36.1+/-15.1 (13 to 73) years at the time of surgery. Pedicle screws were inserted using anatomic landmarks; specific entry sites, preoperative anteroposterior/lateral radiographs, and CT imaging were used to guide the surgeon. After preparation of entry point, a pedicle probe was carefully advanced free hand down the pedicle into the body. Careful palpation of all bony borders (flour and 4 pedicle walls) was performed before placement the screw. Postoperative CT scans were used to evaluate the position of all pedicle screws inserted. Screw cortical penetration was graded on the basis of anatomy (the wall penetrated) and distance of the penetration, with 2 mm of interval. RESULTS: The number of screws inserted at each level were as follows: T10 (n=55), T11 (n=124), T12 (n=118), L1 (n=91), L2 (n=146), and L3 (n=106). Analysis of these pedicle screws using postoperative CT scans confirmed 37 (5.8%) violated screws, including 12 medial, 18 lateral, 2 superior, 1 inferior, and 4 anterolateral vertebral body penetrations. No neurologic, vascular, or pleural injuries occurred. No screws required postoperative repositioning. CONCLUSIONS: Pedicle screw placement with a free hand technique after reviewing preoperative imaging seems to be accurate, reliable, and safe adjunct for the placement of thoracolumbar spine screws. PMID- 18418140 TI - Thoracic myelopathy due to ossification of the yellow ligament in young baseball pitchers. AB - STUDY DESIGN: Case series. OBJECTIVES: To report rare cases of thoracic myelopathy due to ossification of the yellow ligament (OYL) in relatively young baseball pitchers and show clinical evidence of the role of dynamic mechanical stress on the development of OYL. SUMMARY OF BACKGROUND DATA: The pathogenesis of OYL is still unclear. The majority of cases of OYL occur in middle-aged men whereas younger people are rarely affected. This has lead to the hypothesis that diffuse mechanical stress and degenerative changes correlate with the development of OYL. However, there have been no clinical reports demonstrating the critical role of mechanical stress in the ossification. METHODS: Two young highly active baseball pitchers with thoracic myelopathy due to OYL are presented. Both had no previous systemic disorders or family history of treatment for OYL. Magnetic resonance imaging and computed tomography demonstrated compression of the spinal cord by unilateral left sided OYL at the level of the thoracolumbar junction. RESULTS: Both patients were treated with posterior decompression. They recovered full muscle power after operation and resumed pitching training. CONCLUSIONS: Patients had no other factors influencing the development of OYL and the lesions were localized at the left side in the thoracolumbar junction, indicating that repeated, localized rotatory mechanical stress caused by the pitching motion probably influenced the development of OYL in these young baseball pitchers. PMID- 18418141 TI - Intraoperative localization of thoracic spine level with preoperative percutaneous placement of intravertebral polymethylmethacrylate. AB - OBJECTIVE: To evaluate the safety and utility of preoperative vertebroplasty for intraoperative localization of thoracic spinal levels. SUMMARY OF BACKGROUND DATA: Intraoperative fluoroscopy or plain radiographs are traditionally used to localize thoracic spine levels during thoracic spine operations. Unfortunately, such localization can occasionally be difficult in the midthoracic levels due to lack of landmarks, scapular shadows, and the body habitus of the morbidly obese. There are multiple techniques described in the literature that allow for preoperative localization of thoracic spinal levels during approaches to the posterior thoracic spine. For efficient and accurate intraoperative localization of thoracic spinal levels during anterior thoracic spine procedures, we describe a method that uses preoperative percutaneous placement of polymethylmethacrylate (PMMA) into the vertebral body using standard vertebroplasty technique. METHODS: Four patients with morbid obesity and symptomatic thoracic disc herniations underwent preoperative vertebroplasty procedures using standard percutaneous techniques. The PMMA cement was used to expeditiously identify thoracic spinal levels of interest using intraoperative fluoroscopy. RESULTS: All 4 patients underwent successful vertebroplasty procedures without complications. The PMMA cement was easily identified intraoperatively and led to the correct identification of the thoracic spinal levels of interest. CONCLUSIONS: Preoperative placement of PMMA into thoracic vertebral bodies using standard vertebroplasty technique provides a safe, efficient, and reliable method of localizing thoracic spine levels intraoperatively. Such procedures can be performed in the outpatient setting and can be associated with extremely low morbidity when done by experienced practitioners. This procedure should be reserved for patients in whom a surgeon anticipates difficulty using standard radiographs or fluoroscopy to localize thoracic spinal levels intraoperatively. PMID- 18418142 TI - An essential principle for safe C1-2 transarticular screw insertion. PMID- 18418155 TI - What you should know about drug compatibility. PMID- 18418159 TI - Administering an enoxaparin injection. PMID- 18418160 TI - Responding to neurologic effects of Lyme disease. PMID- 18418161 TI - Myths and facts... About shaken baby syndrome. PMID- 18418167 TI - Doing right to prevent wrong-site surgery. PMID- 18418176 TI - Watercolors in the rain. PMID- 18418177 TI - Getting a grip on stress. PMID- 18418178 TI - Seven legal tips for safe nursing practice. AB - Use these pointers to help provide safe patient care and reduce the risk of lawsuits. PMID- 18418180 TI - Raising the SBAR: how better communication improves patient outcomes. PMID- 18418181 TI - Hard to swallow: understanding dysphagia. AB - Review what causes this potentially dangerous disorder and how to help patients who struggle with it. PMID- 18418182 TI - Caring for a patient after coronary artery bypass graft surgery. AB - Follow this system-by-system approach to keeping your patient stable and steering clear of complications. PMID- 18418184 TI - Smooth patient transfers, part III: using a hydraulic lift for patient transfer. AB - Learn how this "pick-me-up" can protect your patient and your back. PMID- 18418187 TI - Detecting women at risk for postpartum mood disorders. PMID- 18418193 TI - Promoting safe pediatric resuscitation. PMID- 18418195 TI - Contrast reaction. PMID- 18418196 TI - Pharmacogenetics and pharmacogenomics for the infectious diseases practitioner. PMID- 18418198 TI - Improving nursing practice in the NICU. PMID- 18418199 TI - Protecting ourselves and others from influenza. PMID- 18418200 TI - Validation of the N-PASS. PMID- 18418201 TI - Endotracheal suctioning increases cerebral blood flow in the very low birth weight infant. PMID- 18418202 TI - Hazards of cleaning medical equipment. PMID- 18418203 TI - Feeding milk at the perfect temperature. PMID- 18418204 TI - Cobedding: what is the evidence? PMID- 18418207 TI - Humidified high-flow nasal cannula: is it the new and improved CPAP? AB - Humidified high-flow nasal cannula (HHFNC) therapy for neonates has become increasingly popular in recent years. It is frequently used in circumstances when continuous positive airway pressure (CPAP) might otherwise have been used. Limited evidence is available to support the specific role, efficacy, and safety of HHFNC in newborns. This evidence suggests that HHFNC provides inconsistent and relatively unpredictable positive airway pressure, but may be effective in the treatment of some neonatal respiratory conditions while being more user-friendly for caregivers than conventional CPAP. Concerns regarding infection risks have resulted in one major HHFNC device being withdrawn from the market before being recently reintroduced. Caution should be exercised in the use of HHFNC in neonates (eg, by limiting its use to relatively lower flows and to relatively larger neonates, while collecting and closely monitoring safety and efficacy data) until further evidence is available to clearly delineate its role, as well as to support its safety and efficacy. PMID- 18418205 TI - The nursing care of the infant receiving bubble CPAP therapy. AB - There are increasing numbers of low birth-weight and premature infants surviving with conditions such as chronic lung disease or bronchopulmonary dysplasia due to complications of assisted mechanical ventilation and other factors. Continuous positive airway pressure (CPAP) has been used as an alternative respiratory treatment to prevent and manage lung disease in preterm infants since the 1970s. Evidence has demonstrated the usefulness of CPAP in the delivery room, as a rescue therapy, as an extubation tool, and a method for managing apnea of prematurity. Bubble CPAP is a unique, simple, inexpensive way of providing continuous positive pressure to infants. Some background and training in the setup, care, and evaluation of the infant on bubble CPAP is essential for positive outcomes. PMID- 18418208 TI - Congenital diaphragmatic hernia: advanced physiology and care concepts. AB - Congenital diaphragmatic hernia (CDH) is a defect in the formation of the diaphragm of the fetus. The diaphragm is the muscle and tissue that separate the chest and the abdominal cavities. In CDH, abdominal organs push into the chest cavity through the defect or herniation, compressing the developing lungs. During the past 10 years, significant changes have occurred in the diagnosis and management of CDH. Despite advances in postnatal care, infants born with a CDH continue to suffer substantial morbidity and mortality. Healthcare providers continue to research therapeutic approaches that will improve the care and optimize survival in these infants. The purpose of this article is to offer an in depth exploration of neonatal physiology and pathophysiology, providing advanced concepts that expand the scientific basis for neonatal care practices. PMID- 18418209 TI - Nursing Care Guidelines for prevention of nasal breakdown in neonates receiving nasal CPAP. AB - Use of nasal continuous positive airway pressure (CPAP) is increasing as a means of respiratory support for respiratory distress syndrome in many premature neonates across the United States. Nasal CPAP is less invasive and may be as effective as mechanical ventilation in premature infants, and has been shown to cause less lung damage in premature neonates. Because of the increased use of nasal CPAP in neonates, especially younger and more fragile neonates, the presence of nare and nasal septum breakdown may be seen as a complication. Currently, all nasal CPAP systems use a hat and prong or mask type of delivery system. This appears to be effective for many neonates, but for some, it is difficult to appropriately fit the hat and prongs. The result of an inappropriately fitted device can be mild to severe nare and nasal septum damage. This article will discuss the need for nasal CPAP and the mechanics of nasal CPAP, but more importantly, serve to guide caregivers in the appropriate physical assessment and care of a neonate on nasal CPAP with the goal of preventing skin breakdown and nasal damage. PMID- 18418210 TI - Extremely long hospitalizations of newborns in the United States: data, descriptions, dilemmas. AB - PROBLEM: Neonatal and pediatric nurses and physicians care for newborn children who have been saved by technological support but who then spend extremely long periods of time in the hospital, perhaps never being able to be discharged to home. There has been little research identifying newborns who are too sick to be discharged from the health care setting and rare reports of staff or parental response to these long-term hospitalizations. PURPOSE: This study provides both the numerical data and description of acutely, chronically ill newborn children whose illnesses caused hospitalizations for greater than 6-months (179 days) in the US. METHODOLOGY: Method triangulation using a national data set (HCUPKID 2003), a researcher created LONGTERM survey, and a qualitative question was used to identify pathologies associated with newborn length of stays greater than 6 months. Neonatal nurses and physicians provided descriptions of children spending at least 6 months in the hospital, including anecdotal reports of caring for those children. RESULTS: The national H-CUP data set identified 680 infants staying 6 months or longer in the hospital during 2003. Four hundred and twenty two providers submitted LONGTERM surveys describing these infants, with 228 first hand reports on how it felt to care for children with hospital stays between 6 months and 6 years. Extreme prematurity, respiratory distress and necrotizing enterocolitis contributed to the extremely long hospital stays. Nurse and physician participants felt that extremely long hospital stays were often due to situations in which parents or colleagues were insisting upon continued futile treatment. PMID- 18418211 TI - New directions in the management of advanced pancreatic cancer: a review. AB - Complete surgical resection is the only potentially curative option for pancreatic cancer. However, most patients have advanced/metastatic disease at the time of diagnosis, or will relapse after surgery. Systemic chemotherapy is only palliative. Gemcitabine-based therapy is an acceptable standard for unresectable locally advanced/metastatic pancreatic cancer, but average median survival is only 6 months. The addition of other chemotherapies (including other antimetabolites, platinum, and topoisomerase I inhibitors) or targeted therapies (farnesyl transferase inhibitors, metalloproteinase inhibitors, cetuximab and bevacizumab) to gemcitabine has failed to improve outcome. The combination of gemcitabine and erlotinib, a small-molecule tyrosine kinase inhibitor of the human epidermal growth factor receptor, was recently approved by the US/European authorities for use in advanced disease. In a phase III trial, the combination demonstrated a significant improvement in overall survival compared with gemcitabine monotherapy. Positive efficacy results have also been observed in a phase III trial, favoring the addition of capecitabine to gemcitabine compared with gemcitabine alone. This review focuses on the recent developments in systemic treatment, and discusses how novel agents might be incorporated into future treatment strategies for pancreatic cancer. PMID- 18418212 TI - Capecitabine: an overview of the side effects and their management. AB - Xeloda (capecitabine), a thymidine phosphorylase activated fluoropyrimidine carbamate, is currently the only universally approved orally administered 5 fluorouracil (5-FU) prodrug. It belongs to a newer generation of orally administered fluoropyrimidines. It has been developed because of the clinical need for efficient, tolerable and convenient agents, which do not require continuous infusion. Capecitabine is not a cytotoxic drug in itself, but via a three-step enzymatic cascade, it is converted to 5-FU mainly within human cancer cells. While the drug compares favorably with 5-FU in patients with advanced or metastatic colorectal cancer and pretreated breast cancer, it also has an improved toxicity profile, mainly of gastrointestinal and dermatologic effects with a significantly lower incidence of grade 3/4 myelotoxicity compared with infusional 5-FU-based chemotherapy. Capecitabine's selective activation within the tumor allows for less systemic toxicity events. A gradient of fluoropyrimidine toxicity is observed: high in the US and low in East Asia. In addition, there is a discrepancy in tolerance of dose among patients treated in the US vs. Europe. Although patients can take the drug orally in the convenience of their own home, the key to successful management of capecitabine is the clinician's awareness of its severe, but low in incidence, adverse effects, and the patients' education, emphasizing compliance with the treatment plan, prevention and timely recognition of its toxicities. PMID- 18418213 TI - Dose-dependent and sequence-dependent cytotoxicity of erlotinib and docetaxel in head and neck squamous cell carcinoma. AB - The purpose of this study was to determine whether the efficacy of taxoid treatment combined with epidermal growth factor receptor (EGFR) inhibition is dose and sequence dependent in head and neck squamous cell carcinoma. Three head and neck squamous cell carcinoma cell lines, chosen on the basis of their diverse EGFR expression levels, were treated with docetaxel, erlotinib, or both. The combination index was calculated using the Chou-Talalay equation. Propidium iodide staining with fluorescence-activated cell sorting analysis was used to evaluate the effects of drugs on cell cycle changes. Western blot analysis was used to determine the effects of agents on cell signaling pathways. Administration of low-dose docetaxel (0.1-3 nmol/l) concurrently or before erlotinib had additive cytotoxic effects in two cell lines but was antagonistic in one line, whereas low-dose docetaxel after erlotinib was synergistic in all cell lines. In contrast, high-dose docetaxel (40 nmol/l) resulted in more apoptosis when given before, rather than after or concurrently with, erlotinib. Low-dose docetaxel induced an accumulation of cells in the sub-G0 phase of the cell cycle with no mitotic arrest or apoptosis, whereas high-dose docetaxel induced mitotic arrest and apoptosis. The low and high doses of docetaxel had opposite effects on EGFR expression: a decrease and an increase, respectively. The dose of docetaxel affects sequence-dependent cytotoxicity when docetaxel is combined with an EGFR inhibitor. The mechanism for this difference is a combination of the dose-dependent effects of docetaxel on the mode of cell death and on EGFR expression. PMID- 18418214 TI - Inhibition of gamma-secretase affects proliferation of leukemia and hepatoma cell lines through Notch signaling. AB - Notch signaling is a well-conserved pathway playing crucial roles in regulating cell fate decision, proliferation, and apoptosis during the development of multiple cell lineages. Aberration in Notch signaling is associated with tumorigenesis of tissues from various origins. To investigate the role Notch signaling plays in the proliferation of cancer cell lines, the expression profiles of Notch1 in six human cancer cell lines (Jurkat, HepG2, SW620, KATOIII, A375, BT474) were examined. All cell lines differentially expressed Notch1, and only Jurkat and SW620 expressed cleaved Notch1 (Val1744). Among the six cell lines tested, only Jurkat and HepG2 showed a decrease in cell proliferation during 4 days of treatment with a gamma-secretase inhibitor (GSI). This is the first report on the anti-proliferative effects of GSI on a human hepatoma cell line. These two cell lines expressed Notch1-3, Jagged1, Jagged2, Dlk1 and Hes1. GSI treatment led to a decrease in Hes1 expression in both cell lines. Surprisingly, GSI treatment resulted in the accumulation of Notch1 protein upon treatment. During this period, GSI treatment did not induce apoptosis, but caused cell cycle arrest in both cell lines. This was also correlated with decreased c myc expression. Forced expression of activated intracellular Notch1 completely abrogated GSI sensitivity in both cell lines. These results clearly demonstrate that Notch signaling positively regulates cell proliferation in Jurkat and HepG2 cell lines and that GSI treatment inhibits tumor cell proliferation through the suppression of Notch signaling. PMID- 18418215 TI - Nordihydroguaiaretic acid restores expression of silenced E-cadherin gene in human breast cancer cell lines and xenografts. AB - In our study we use nordihydroguaiaretic acid (NDGA), the naturally occurring lignan, to investigate whether it plays a role in the prevention and treatment of cancer by epigenetic modifications. The growth inhibitory effect of NDGA on human breast cancer cell lines was determined using the MTT assay (3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay). It substantially inhibited the growth of human breast cancer cell lines SKBR3 and MDA-MB-435 with an estimated IC50 of 31.09+/-1.6 and 38.8+/-2.1 micromol/l respectively, after 4 days incubation with different NDGA concentrations. The in-vivo anticancer activity of NDGA was evaluated by calculating the tumor growth inhibition value. NDGA substantially inhibited the growth of human breast carcinoma cells in both animal and cell-based models. We also found that a single treatment with NDGA reactivates methylation-silenced E-cadherin gene in vitro and in vivo, suggesting an intriguing concept that lignans may act as natural effective epigenetic modifiers in the prevention and treatment of cancer. PMID- 18418216 TI - Hyperbilirubinemia's protective effect against cisplatin nephrotoxicity in the Gunn rat. AB - Gunn rats, deficient in the enzyme uridine diphosphate glucuronyl transferase, were used to investigate the effects of unconjugated hyperbilirubinemia in cisplatin nephrotoxicity. The effect of bilirubin on the antineoplastic activity of cisplatin in osteosarcoma cell lines was also determined. The in vivo model involved three groups of rats (n=6 rats/group): homozygous Gunn rats (j/j), heterozygous Gunn rats (j/+), and congenic Wistar rats. On day 0, all rats were given 4 mg/kg cisplatin intraperitoneally. Blood was sampled on days 0, 3, and 5 for bilirubin, BUN, and creatinine and kidneys were taken on day 5. Cell culture was performed in four canine osteosarcoma cell lines using the average concentrations of bilirubin for homozygous Gunn rats at day 0 and 3. Bilirubin was added to cell lines alone and with cisplatin. Cell viability was assessed using the CellTiter Blue assay. Serum bilirubin levels were highly elevated in Gunn j/j, moderately elevated in Gunn j/+, and undetectable in Wistar rats at day 0. Bilirubin provided a nephroprotective effect, with significantly lower BUN and creatinine in Gunn j/j when compared with Wistar rats at day 5. Histological grading demonstrated preservation of the S3 segment in Gunn j/j when compared with Wistar rats (P<0.05). Bilirubin had no significant effect on the antineoplastic effect of cisplatin at either concentration in the four cell lines (P<0.001). Hyperbilirubinemia in the Gunn rat provided marked preservation of renal function and histology in a cisplatin nephrotoxicity model. Exogenous bilirubin did not interfere with the antineoplastic activity of cisplatin in vitro. PMID- 18418217 TI - Gene induction by glycyrol to apoptosis through endonuclease G in tumor cells and prediction of oncogene function by microarray analysis. AB - Glycyrrhiza uralensis (Leguminosae) has long been known as an antiinflammatory agent for gastric ulcers, arthritis, and rheumatism. The flavonoid glycyrol (GC) (10 microg/ml) isolated from G. uralensis dramatically inhibits phorbol ester (phorbol 12-myristate 13-acetate)-induced nuclear factor (NF)-kappaB-dependent transcriptional activity, as determined by luciferase reporter activity in human kidney epithelial 293T cells. To investigate global gene expression profiling in cells by GC, we performed high-density oligonucleotide microarrays. Our microarray analyses showed that GC inhibited phorbol ester-induced NF-kappaB dependent transcriptional activity in inflammatory-related gene expression. RT PCR analysis, based on microarray data, showed that NF-kappaB-dependent genes (such as CCL2, CCL7, CD44, and HSPB8 in addition to NF-kappaB itself) were significantly downregulated by GC. Treatment with GC (10 microg/ml) inhibited I kappaB degradation induced by phorbol 12-myristate 13-acetate. The microarray data also suggested that GC induces gene expression to p53-dependent apoptosis through endonuclease G, instead of CAD/DFF and AIF/PDCD8, as a downstream apoptosis factor in human kidney epithelial 293T tumor cells, and induces oncogenes with a suppressor role as an added function. PMID- 18418218 TI - Potentiation of anticancer-drug cytotoxicity by sea anemone pore-forming proteins in human glioblastoma cells. AB - The search for new drugs and treatment approaches is of particular importance for glioblastomas (GBMs), as with other types of malignant gliomas, as they are lethal without the available medical care. Current anticancer cocktails have failed to prolong survival beyond 1 year, in part owing to the natural resistance of GBM cells and to the toxic side effects of the available drugs. In many organisms, cell death can be induced by cytolysins, which are proteins that can form pores in biological membranes. Perhaps by facilitating drugs to enter into the cytosol, cytolysins might be used to increase the efficacy of conventional anticancer agents. Here, the cytotoxicity of two sea anemone pore-forming cytolysins, toxin Bc2, and equinatoxin (EqTx-II) were investigated. Toxin Bc2 and EqTx-II were cytotoxic against human U87 and A172 GBM cell lines either wild type or p53 mutant, a tumor suppressor frequently mutated in malignant gliomas. Moreover, noncytotoxic concentrations of Bc2 or EqTx-II potentiated the cytotoxicity induced by low dose concentrations of all classical chemotherapeutics agents tested: cytosine arabinoside, doxorubicin, and vincristine. In comparison with the cytotoxicity induced by each of these classical anticancer drugs alone, 10-300-fold less of the therapeutic drug was needed when combined with the cytolysins. These results are promising, since lower concentrations of chemotherapeutic drugs could reduce the adverse effects of chemotherapy. PMID- 18418219 TI - Interferon-alpha in combination with either imatinib (Gleevec) or gefitinib (Iressa) in metastatic renal cell carcinoma: a phase II trial. AB - Treatments for metastatic renal cell carcinoma (MRCC) are limited. RCCs frequently overexpress epithelial growth factor receptor and express c-Kit and platelet-derived growth factor receptor-beta. Combination of interferon with tyrosine kinase inhibitors of epithelial growth factor receptor [gefitinib (Iressa)] or c-Kit and platelet-derived growth factor receptor-beta [imatinib (Gleevec)] was evaluated for efficacy and safety. Patients with MRCC received 12 week cycles of interferon [3 million units (MU) subcutaneously thrice in week 1 and 6 MU thrice weekly thereafter] and either gefitinib (500 mg daily) or imatinib (600 mg daily). The gefitinib/imatinib dose was reduced as needed owing to toxicity. The primary endpoint was objective tumor response. Secondary endpoints were time to tumor progression, overall survival, and safety. Seventeen patients were enrolled. Most had clear cell [36% (6/17)] or papillary [36% (6/17)] tumors. Most (n=14) were treated on the gefitinib arm, including two patients who crossed over from the imatinib arm after experiencing disease progression. Objective tumor responses were evaluable in 14 patients (82%). Of these 14, partial responses occurred in three (21%), stable disease in seven (50%), and progressive disease in four (29%). The most frequent treatment-related adverse events were skin rash, flu-like symptoms, and fatigue (both treatment arms); diarrhea (gefitinib arm only); and thrombocytopenia and leukopenia (imatinib arm only). Median time to tumor progression (range) for patients on the gefitinib arm only was 4.27 (1.13-15.97) months and median overall survival (range) was 11.42+ (1.13-29.07+) months. Combination of gefitinib with interferon safely delays progression of refractory MRCC. Further studies in this setting are warranted. PMID- 18418220 TI - Phase II study of docetaxel in combination with oxaliplatin in patients with metastatic or locally advanced esophagogastric cancer previously untreated with chemotherapy for advanced disease: results of the Central European Cooperative Oncology Group Study ESGAS.1.2.001. AB - A phase II trial was performed to determine the efficacy and tolerance of docetaxel plus oxaliplatin with hematopoietic growth factor support in previously untreated patients with advanced gastroesophageal adenocarcinoma. Thirty-five patients were entered in this trial. Treatment consisted of 3-weekly docetaxel 80 mg/m2 and oxaliplatin 100 mg/m2 both infused on day 1. A prophylactic 5-day course of human granulocyte colony-stimulating factor 5 microg/kg/day was given subcutaneously, and erythropoietin (10,000 IU subcutaneously three times per week) was administered if hemoglobin was less than 12.0 mg/dl. The confirmed overall response rate was 34%, including two complete responses (6%) and 10 partial responses (28%). Fifteen patients (43%) had stable disease. The median time to response was 2.5 months (1-3.5), the median time to progression was 8.9 (4-42.5) months and the median overall survival time was 11.6 (2.5-51) months. Hematologic toxicity was common, though World Health Organization grade 3 or 4 neutropenia occurred only in six (17%) patients and anemia in six (17%) patients, respectively. Nonhematologic adverse reactions were usually mild-to-moderate. Our data suggest that the combination of docetaxel and oxaliplatin with granulocyte colony-stimulating factor and erythropoietin has a promising therapeutic index in patients with advanced gastroesophageal adenocarcinoma. PMID- 18418221 TI - Phase II trial of biweekly pegylated liposomal doxorubicin in recurrent platinum refractory ovarian and peritoneal cancer. AB - Pegylated liposomal doxorubicin (PLD) is active in recurrent platinum-refractory ovarian and peritoneal cancer as demonstrated in a prospective-randomized trial. Dose-limiting toxicity in the US Food and Drug Administration-approved application schedule of PLD (50 mg/m2 every 4 weeks) was serious palmar-plantar erythrodysaesthesia (PPE). This phase II trial was aimed at reduction of the frequency of serious PPE without loss of efficacy by modifying both the application schedule and the total dose of PLD administered as palliative single agent chemotherapy. Fifty patients were enrolled into this phase II trial. PLD was given via 30-min intravenous infusion at a dose of 20 mg/m2 every 2 weeks. Primary endpoint of the trial was the best response to chemotherapy. Secondary goals were progression-free survival, overall survival, and toxicity. Four complete responses and 16 partial responses were found resulting in an overall best response rate of 40.0%. The median progression-free survival in the intention-to-treat-population was 4.1 months [95% confidence interval (CI), 2.8 5.4 months], whereas the overall survival was 16.5 months (95% CI, 12.3-20.7 months). Although 17 (34.0%) patients developed some PPE, only one progressed to grade 3 (NCI-CTC version 2.0; 1998). No grade 4 toxicity occurred. PLD 20 mg/m2 biweekly is highly active in patients with recurrent platinum-refractory ovarian and peritoneal cancer. The frequency of grade 3 and grade 4 PPE was remarkably reduced in this trial, as compared with the frequency of serious PPE observed in patients who were administered the dose schedule of 50 mg/m2 every 4 weeks. PMID- 18418222 TI - Sustained clinical responses to tyrosine kinase inhibitor sunitinib in thyroid carcinoma. AB - The limited therapeutic options available for patients with metastatic papillary thyroid carcinomas (PTC) and follicular thyroid carcinomas (FTC) necessitates the development of novel therapies. Identification of somatic rearrangements of the tyrosine kinase domain of the RET gene in PTC have improved our understanding of thyroid tumorigenesis. Sunitinib is active against the RET kinase and has both antineoplastic and antiangiogenic properties. Its role in the treatment of patients with thyroid carcinoma has yet to be evaluated in clinical trials. Two patients with progressive metastatic thyroid carcinoma (case 1: PTC, and case 2: FTC) were enroled in a phase I clinical trial to evaluate positron emission tomography (PET) in the monitoring of response to sunitinib. Tumour biopsies and PET were performed at baseline and 4 weeks after the commencement of sunitinib. Activation of the RET kinase pathway was evaluated using immunohistochemistry (IHC) and western blot analysis of total phosphorylated tyrosine and downstream signalling targets of the RET pathway. Both patients demonstrated sustained clinical responses to sunitinib over a duration of 4 years. In case 1, (PTC) PET confirmed evidence of a partial metabolic response, and IHC and western blot analysis demonstrated inhibition of the RET kinase pathway posttreatment. In case 2, (FTC) PET confirmed stable disease after sunitinib. IHC staining of the tumour showed low total phosphorylated tyrosine staining at baseline which did not change after treatment. These case studies highlight potential activity of sunitinib in patients with metastatic thyroid carcinoma. Sunitinib seems to be a promising agent in the treatment of thyroid cancers and this requires validation in future clinical trials. PMID- 18418223 TI - Managing cancer pain: practical principles and pearls for pragmatic polypharmacy. PMID- 18418224 TI - Adjuvant agents in cancer pain. AB - Cancer-related neuropathic pain derives from peripheral or central lesions of the nervous system and is often associated with the hallmark symptoms of allodynia (pain from a stimulus that does not normally evoke pain) and hyperalgesia (an exaggerated pain response to a normally painful stimulus). Pain is prevalent in patients with cancer and considerably undermines their quality of life, thereby making the development of a comprehensive pain management approach essential. Coanalgesics have been well integrated into cancer pain management strategies and are often used as first-line options for treatment of certain disease processes such as neuropathic pain. These medicines, including antidepressant and anticonvulsant agents, are recommended by evidence-based guidelines, whereas others, such as lidocaine patch 5%, are supported by randomized, controlled, clinical trial data. In addition to understanding which agents are recommended for neuropathic pain, it is useful to know which agents are of limited utility or are to be avoided when prescribing treatment for neuropathic pain. Notwithstanding the need for head-to-head studies before firm statements on comparative efficacy can be made, it is worth considering the numbers needed to treat statistic for the treatment of neuropathic pain with coanalgesics. Potentially harmful treatments are considered, as well as the numbers needed to treat, mechanisms of action, and clinical trial data for agents that can be beneficial for the management of cancer-associated neuropathic pain. PMID- 18418225 TI - Current developments in opioid therapy for management of cancer pain. AB - Pain remains a highly prevalent problem for patients with cancer and typically falls into one of 3 types: visceral, somatic, and neuropathic. A mechanistic, pathophysiologic approach to pain management involves a good assessment of the type of pain, followed by tailoring of the treatment based on the diagnosis. This pain management strategy can provide rapid pain control with a lower incidence of complications and side effects than other methods. Furthermore, pharmacogenetics may play an important role in individualizing therapies in the future, but for now this type of data offers explanations for phenomena commonly observed in clinical practice, such as (1) differences in individual analgesic and side effect responses to various opioid agents, (2) incomplete cross-tolerance seen when switching between mu opioid analgesics, and (3) why opioid rotation can be beneficial for patients after an opioid therapy loses efficacy or becomes associated with intolerable side effects. Especially for difficult-to-manage pain patients, additions to the opioid analgesic armamentarium can potentially better individualize pain management, and provide another option to be used for opioid rotation. Among the most recent Food and Drug Administration-approved opioid analgesics for acute pain and persistent pain are oral immediate-release and extended-release formulations of oxymorphone, whereas for breakthrough pain, the ultrarapid-acting opioid, fentanyl effervescent buccal tablets, has newly been developed and indicated within the United States. PMID- 18418226 TI - Management of expected and unexpected opioid-related side effects. AB - Intolerable side effects contribute to poor outcomes among patients managed with opioids, and negotiating these side effects remains an important clinical challenge. Evidence-based approaches to minimizing the side-effect burden from opioids compiled from the literature, and including clinical practice recommendations, focus on (1) reducing the doses of systemic opioids, (2) managing the adverse symptoms of opioids, (3) integrating opioid rotation, and (4) changing the route of administration. These broad strategies, and specific recommendations for individual side effects of opioids, including those that often go unrecognized within the following organ systems: neurologic, cardiopulmonary, gastrointestinal, urologic, endocrinologic, dermatologic, and immunologic, are considered herein. In addition, specific patient populations, opioid metabolites, and clearance issues are considered. In particular, novel medications and better alternatives to preexisting treatments for specific opioid side effects, such as peripheral opioid antagonists, may ameliorate the management of patients taking long-term opioid therapies. PMID- 18418227 TI - Lipoprotein(a) is associated with coronary heart disease independent of metabolic syndrome. AB - AIM: To assess (i) the association between lipoprotein(a) [Lp(a)] with the likelihood of coronary heart disease and metabolic syndrome (MS) and (ii) its covariates in Turkish adults. METHODS: Cross-sectional evaluation of 1309 adults, who had serum Lp(a) determinations by Behring nephelometry, and followed for a mean 1.0 year. MS was defined by ATPIII criteria modified for male abdominal obesity. RESULTS: Mean age of the sample was 56.8+/-11.3 years. After adjustment for sex, age, and smoking status, log-transformed Lp(a) levels were associated significantly with coronary heart disease likelihood in both sexes combined [odds ratio: 1.53 (95% confidence interval: 1.06; 2.20)]. This association persisted after additional adjustment for MS [odds ratio: 1.57 (95% confidence interval: 1.09; 2.26)]. The Lp(a) mid-tertile (5-17 mg/dl), accompanied by significantly lower serum triglycerides than the two remaining tertiles, was inversely associated significantly with MS in either sex; in women, this association was independent of waist circumference. In a linear regression comprising seven variables, excepting total cholesterol, only gamma-glutamyltransferase in women (P=0.002) and waist circumference (P=0.057) in men were inverse covariates of modest magnitude of Lp(a). CONCLUSION: Coronary heart disease likelihood, significantly associated with Lp(a) concentrations, is independent of MS and insulin resistance. Suggestive evidence was provided that intermediary Lp(a) concentrations, when accompanied by the presence of MS, could accelerate progression of vascular disease, especially in women. PMID- 18418228 TI - Th2-dependent cytokine release in patients treated with coronary angioplasty. AB - AIM: The aim of this study is to investigate the T helper (Th)2-dependent release of interleukin(IL)-4 and IL-13 in patients with unstable angina treated with coronary angioplasty (PCI). METHODS: This study involved 48 consecutive patients with unstable angina who underwent PCI. Blood samples were taken before and 48 h after PCI to evaluate serum IL-4 and IL-13 levels. Coronary atherosclerosis was assessed in terms of the severity of the treated lesions and the presence of multivessel disease, and compared with IL release. RESULTS: Before and after PCI, serum IL-4 levels were, respectively, 53.1+/-110.7 and 35.1+/-16.9 pg/ml (P not significant), and serum IL-13 levels 6.7+/-3.7 and 6.0+/-2.9 pg/ml (P<0.05). A correlation was present between the severity of the treated coronary lesions and serum IL-13 levels (r 0.36; P<0.05). The patients with multivessel coronary artery disease had a higher periprocedure ratio of IL-4 than those with single vessel coronary artery disease (1.46+/-0.76 vs. 0.89+/-0.34, P<0.001). CONCLUSION: A significant decrease was observed in Th2-dependent IL concentrations after PCI; increased Th2-dependent IL levels before PCI seem to correlate with the severity of coronary atherosclerosis. PMID- 18418229 TI - Endothelial dysfunction and carotid lesions are strong predictors of clinical events in patients with early stages of atherosclerosis: a 24-month follow-up study. AB - BACKGROUND: The purpose of this study was to investigate whether the vasodilator response to brachial artery and the presence of carotid lesions may have a prognostic significance in patients with early stages of atherosclerosis. METHODS AND RESULTS: Vascular echography was performed to analyze flow-mediated vasodilatation (FMD) at the brachial artery and intima-media thickness (IMT) of carotid arteries in 84 asymptomatic patients. At baseline, we evaluated all the established traditional cardiovascular risk factors. Transient ischemic attack, stroke, effort or unstable angina, acute myocardial infarction, peripheral arterial disease and cerebrovascular and cardiovascular death served as outcome variables over a follow-up period of 24 months. Brachial FMD was correlated inversely with carotid IMT (P=0.003), systolic blood pressure (P=0.0001) and age (P=0.0001). IMT was positively correlated with systolic blood pressure (P=0.0001), waist circumference (P=0.004) and age (P=0.01). At the end of the follow-up cardiovascular and cerebrovascular events were registered in 29% of the patients and in a multivariate analysis, including all the variables evaluated at baseline, male sex [odds ratio (OR) 1.6, P=0.005], the presence of baseline carotid lesions (OR 3.5, P=0.02) and FMD below the median (OR 3.2, P=0.03) were the only variables predictive of clinical events. CONCLUSION: In this study, endothelial dysfunction and carotid lesions significantly increased the risk of vascular events in asymptomatic patients with early stages of atherosclerosis. Assessment of systemic vasoreactivity and carotid IMT evaluation may provide, in this category of patients, important prognostic information in addition to that derived from traditional established cardiovascular risk factors. PMID- 18418230 TI - Postprandial hypertriglyceridemia associated with inflammatory response and procoagulant state after a high-fat meal in hypertensive patients. AB - OBJECTIVE: To investigate the changes of inflammatory factors and hemostatic variable in plasma after a high-fat meal in normocholesterolemic patients with essential hypertension. METHODS: A total of 60 hypertensive patients were randomly assigned to accept a single high-fat meal (group 1, n=40) or not (group 2, n=20) in the morning after an overnight fast, and 20 healthy participants (group 3) consumed a single high-fat meal on the same day. Plasma lipid profiles, high-sensitivity C-reactive protein (hsCRP), tumor necrosis factor alpha (TNFalpha), soluble P-selectin and plasminogen activator inhibitor type 1 (PAI-1) antigen levels were measured at fasting and 4 h after meal ingestion. RESULTS: Postprandial triglyceride levels increased significantly in groups 1 and 3 (P<0.01), whereas levels were higher in group 1 (P<0.001). Postprandial plasma TNFalpha, hsCRP, soluble P-selectin and PAI-1 antigen levels increased in group 1 (P<0.001) but not in group 3. Postprandial plasma triglyceride level was correlated with log(hsCRP) (P<0.001), TNFalpha (P<0.001), soluble P-selectin (P<0.01) and PAI-1 antigen (P<0.05) levels, respectively. Both postprandial plasma level of soluble P-selectin and that of PAI-1 antigen were positively and significantly correlated with those of log(hsCRP) (P<0.01) and TNFalpha (P<0.001), respectively. CONCLUSION: Postprandial hypertriglyceridemia in hypertensive patients is associated with inflammatory response and procoagulant state. PMID- 18418231 TI - Homocysteine predicts adverse clinical outcomes in unstable angina and non-ST elevation myocardial infarction: implications from the folate intervention in non ST elevation myocardial infarction and unstable angina study. AB - BACKGROUND: Unstable angina and non-ST elevation myocardial infarction (NSTEMI) are common acute coronary events. Homocysteine is a novel risk factor for coronary heart diseases. Together with the conventional risk factors, they may affect the outcome of non-ST coronary events. OBJECTIVE: This study aims to determine the effect of clinical risk factors that are responsible for the occurrence of mortality, and the composite outcome of mortality, nonfatal myocardial infarction and serious rehospitalization within 6 months after the onset of non-ST acute coronary syndromes. METHODS: A total of 124 Filipino patients were interviewed and tested for blood homocysteine levels and lipid profiles. Outcomes were assessed after 6 months. RESULTS: Homocysteinemia (>16 micromol/l) is associated with increased mortality and composite outcomes (mortality, nonfatal reinfarction, and serious rehospitalization), even if adjusted for conventional risk factors. No association was detected for the conventional risk factors. Earlier acute coronary syndrome was found to be positively associated with mortality and the composite outcomes. Early stroke is associated with increased composite outcomes, whereas greater mortality and adverse outcomes were observed in NSTEMI compared with intermediate-risk unstable angina. CONCLUSION: Increased homocysteine level is associated with mortality and serious nonfatal outcomes in patients with unstable angina and NSTEMI. PMID- 18418232 TI - Association of elevated plasma homocysteine levels with impaired ST-segment resolution after fibrinolytic therapy in acute ST-elevation myocardial infarction. AB - OBJECTIVE: Elevated plasma homocysteine levels are associated with increased intracoronary thrombus burden in acute coronary syndromes. The relationship between plasma homocysteine levels and ST-segment resolution (STR) after fibrinolytic therapy has, however, not been investigated. We sought to investigate this issue. METHODS: Forty-nine patients who were administered fibrinolytic therapy for ST-segment elevation myocardial infarction (STEMI) were included in this study. Maximum STR was measured after 90 min of fibrinolytic therapy. Patients with a maximum STR of less than 50% were included in group 1. Patients with a maximum STR >or=50% were included in group 2. Blood samples were collected from the antecubital vein upon arrival to the hospital. The plasma was separated and kept at -70 degrees C until the time of analysis. Total plasma homocysteine level was determined by the high-performance liquid chromatography method with fluorescence detection. RESULTS: Fifteen patients in group 1 and 34 patients in group 2 were present. Baseline characteristics were similar in both groups. The plasma homocysteine levels were 22.5+/-10 micromol/l in group 1 and 14.1+/-4 micromol/l in group 2 (P<0.001). CONCLUSION: Elevated plasma homocysteine levels are associated with impaired STR after fibrinolytic therapy in acute STEMI. PMID- 18418233 TI - Subclinic atherosclerosis and cardiovascular risk factors in HIV-infected children: PERI study. AB - OBJECTIVES: To compare carotid intima-media thickness (cIMT) of children and adolescents with and without HIV infection and to determine associations among independent socio-demographic, clinical or cardiovascular variables and cIMT in HIV-infected children and adolescents. PATIENTS AND METHODS: This is a matched case-control study comparing 83 HIV-infected and 83 healthy children and adolescents. Clinical and laboratorial parameters, cIMT and echocardiogram were measured. RESULTS: The cIMT was higher in HIV-infected individuals (median 480 microm; interquartile range 463-518 microm) compared with controls (426 microm; range 415-453 microm, P<0.001). In addition, the HIV-infected group showed higher levels of high-sensitive C-reactive protein (medians 1.0 mg/l vs. 0.4 mg/l, P<0.001), glycated hemoglobin (6.1+/-0.9 vs. 5.7+/-0.8%, P=0.028) and triglycerides (medians 0.9 vs. 0.8 mmol/l, P=0.031). Finally, this group showed lower levels of total and high-density lipoprotein-cholesterol. After multivariate analysis, increased cIMT was positively associated with stavudine use [odds ratio (OR): 18.9, P=0.005], left atrial/aorta index (OR: 15.6, P=0.019), suprailiac skinfold (OR: 7.9, P=0.019), tachypnea (OR: 5.9, P=0.031), CD8 lymphocyte count (OR: 5.7, P=0.033) and CD4 T-lymphocyte count (OR: 5.5, P=0.025). cIMT increment was negatively associated with total cholesterol (OR: 0.2, P=0.025) and with CD8 zenith (OR: 0.1, P=0.007). CONCLUSION: In this sample of children and adolescents, having HIV infection was associated with increased cIMT and elevated prevalence of cardiovascular risk factors. These findings suggest that this group should be included in cardiovascular prevention programs. PMID- 18418234 TI - Hypoxia-inducible factor-1alpha induces the coronary collaterals for coronary artery disease. AB - BACKGROUND: Marked variability exists in coronary artery collaterals in patients with ischemic heart disease. Multiple factors are thought to play a role in collateral development; however, the contribution of hypoxia inducible factor 1alpha (HIF-1alpha), which is a transcriptional activator that functions as a master regulator of oxygen homeostasis, is not completely clear. It could play an important role in modulating collateral development. OBJECTIVE: The objective of this study is to investigate the changes and significance of expression of HIF 1alpha in patients with coronary artery collaterals. METHODS: Collateral vessels were determined in 98 patients with >or=70% narrowing of at least one coronary artery without earlier revascularization, 42 patients with coronary artery collaterals and 56 patients with no coronary artery collaterals. Extent of collaterals was expressed as scores according to the Rentrop scoring system. Another 50 cases with normal coronary arteries were selected as control. The levels of HIF-1alpha protein expression in monocyte and lymphocyte in the participants were tested by immunohistochemistry (IHC) and western blot; mRNA levels were measured using reverse transcriptase PCR technique. RESULTS: Compared with the control with normal coronary artery, the patients had higher expression of HIF-1alpha protein tested by IHC and western blot (52.6+/-10.2 vs. 13.7+/-6.2 by IHC, 50.8+/-4.5 vs. 6.5+/-1.8 by western blot); furthermore, significantly higher HIF-1alpha expression was observed in patients with collaterals compared with patients with no collaterals (81.5+/-11.8 vs. 20.7+/-9.4 by IHC; 87.2+/-6.5 vs. 9.5+/-1.4 by western blot). On the transcriptional levels of HIF-1alpha, the result was the same as the protein, there was significant difference of HIF 1alpha between the three groups. The patients with collaterals were the highest (127.3+/-23.9), followed by patients with no collaterals (35.7+/-12.3), and the control were the lowest (23.5+/-9.3). A highly positive correlation was observed between the expression/transcription of HIF-1alpha and collateral score (P<0.01, IHC: r1=0.78, reverse transcriptase PCR: r2=0.69, western blot: r3=0.84). CONCLUSION: These data suggest that higher inductions of HIF-1alpha are associated with coronary collaterals, thus implying that HIF-1alpha may promote coronary collateral formation. Detection of HIF-1alpha expression might be helpful to predict prognosis of patients with coronary artery disease. PMID- 18418235 TI - Total blush score: a new index for the assessment of microvascular perfusion in idiopathic dilated cardiomyopathy. AB - BACKGROUND: The aim of this study was to evaluate tissue-level perfusion in patients with idiopathic dilated cardiomyopathy (IDC), using the myocardial blush grade technique. METHOD: The study population consisted of 26 prospectively enrolled IDC patients (15 women and 11 men; mean age, 59+/-8.8 years) and 26 control subjects (11 women and 15 men; mean age, 54.9+/-10.6 years), whose angiographic films were technically adequate for myocardial blush grade analysis. After grading, we measured total blush score (TBS) for both groups. TBS was determined as the sum of the blush grades of each coronary territory. RESULTS: A total of 156 coronary territories in both groups were assessed. Average of TBS was significantly lower in patients with IDC than in control group (7.6+/-1.2 vs. 8.8+/-0.4; P<0.0001). The TBS significantly and inversely correlated with New York Heart Association class, heart rate, left ventricular end-systolic dimension, and left ventricular end-diastolic pressure, and positively correlated with left ventricular ejection fraction (r=-0.76, P<0.001; r=-0.61, P=0.001; r= 0.77, P<0.0001; r=-0.68, P<0.0001; and r=0.67, P<0.0001, respectively). CONCLUSION: In IDC, decreased TBS might be assumed to be a surrogate marker for a diseased microvascular network in the catheterization laboratory. The relationship between reduced TBS and IDC severity suggests that this index might have prognostic significance. PMID- 18418236 TI - Usefulness of combined high-sensitive C-reactive protein and N-terminal-probrain natriuretic peptide for predicting cardiovascular events in patients with suspected coronary artery disease. AB - OBJECTIVE: We examined whether the combined use of high-sensitive C-reactive protein (hsCRP) and N-terminal-probrain natriuretic peptide (NT-proBNP) could increase the predictive value for future cardiovascular events. BACKGROUND: hsCRP and NT-proBNP both have been shown to be strong predictors of cardiovascular events in patients with coronary artery disease. Few data are, however, available to assess whether combined use of these two distinct biomarkers improves the risk stratification in predicting cardiovascular events. METHODS: A total of 205 participants with suspected coronary artery disease referred for coronary angiography were enrolled in the study. Plasma levels of hsCRP and NT-proBNP were measured before coronary angiography. Cox regression analyses were conducted for the 205 participants, with cardiovascular events being defined as nonfatal myocardial infarction, percutaneous coronary intervention, coronary artery bypass grafting, and ischemic stroke. RESULTS: All patients were divided into four groups by using median values of hsCRP (1.1 mg/l) and NT-proBNP (472.6 fmol/ml): group 1, low hsCRP/low NT-proBNP (n=60); group 2, high hsCRP/low NT-proBNP (n=42); group 3, low hsCRP/high NT-proBNP (n=42); and group 4, high hsCRP/high NT proBNP (n=61). During a median follow-up of 4 years, there were 84 cardiovascular events (41%): 11 events (18%) in group 1, 13 events (31%) in group 2, 20 events (48%) in group 3, and 40 events (66%) in group 4 (P<0.001). Patients with cardiovascular event had significantly attenuated flow-mediated vasodilation (3.6+/-3.4 vs. 5.3+/-3.5%, P=0.001) and increased plasma levels of NT-proBNP (627+/-330 vs. 458+/-196 fmol/ml, P<0.001). Simple linear regression analysis on all studied participants demonstrated significant associations between levels of hsCRP and NT-proBNP (hsCRP vs. NT-proBNP: r=0.354, P<0.001). Cox regression hazards model showed that combined use of NT-proBNP and hsCRP significantly increased predictive value for future cardiovascular events [hazard ratio (HR) 4.922, 95% confidence interval (CI), 2.519-9.617; P<0.0001 for high hsCRP/high NT proBNP vs. low hsCRP/low NT-proBNP]. CONCLUSION: These findings demonstrated that a simple combination of distinct biomarkers of hsCRP and NT-proBNP might provide additional information for predicting cardiovascular events. PMID- 18418237 TI - Prevalence of subclinical atherosclerosis in asymptomatic diabetic patients by 64 slice computed tomography. AB - OBJECTIVES: Patients with diabetes have a 2-fold to 4-fold higher risk of a cardiovascular event than nondiabetic patients. Thus there is a need to identify patients with diabetes who are at risk of cardiovascular events before the onset of symptoms. We studied the prevalence of coronary artery disease in asymptomatic diabetic patients compared with asymptomatic nondiabetic patients by 64-slice computed tomography (CT). METHODS: From 425 asymptomatic patients with coronary risk factors but without known coronary artery disease who underwent 64-slice CT, we identified 93 asymptomatic diabetic patients (diabetic group) and 93 age matched and sex-matched asymptomatic nondiabetic patients. RESULTS: Clinical characteristics were not significantly different between the two groups. Total coronary calcium score was significantly higher in diabetic group than that in nondiabetic group (median 117 vs. 53.5, P<0.0001). No coronary calcium was detected in 30.0% of nondiabetic group compared with 15.1% of diabetic group (P=0.0022). Coronary calcium score more than 400 was detected in 9.7% of nondiabetic group compared with 36.6% of diabetic group (P<0.0001). Coronary plaques were found in 67.7% of nondiabetic group compared with 91.4% of diabetic group (P<0.0001). Multiple plaques were detected in 57.0 and 77.4% of patients in nondiabetic and diabetic group, respectively (P=0.0030). Significant coronary stenosis was found in 16.1% of nondiabetic group compared with 33.3% of diabetic group (P=0.0065). CONCLUSION: Our results show that the prevalence of coronary plaques detectable by 64-slice CT in asymptomatic diabetic patients is very high. PMID- 18418238 TI - Imaging of coronary artery anomalies: the role of multidetector computed tomography. AB - BACKGROUND: Coronary artery anomalies are evaluated by using catheter-based angiography. Multidetector row-computed tomography (MDCT) is a new noninvasive imaging technique that has excellent spatial resolution for detecting the origin and course of a coronary anomalous vessel. OBJECTIVE: To determine the sensitivity of multidetector computed tomography in patients who had coronary artery anomaly demonstrated by conventional coronary angiography. MATERIAL AND METHODS: A retrospective evaluation to identify 23 patients, who underwent retrospective electrocardiographic (ECG)-gated MDCT, was done and in whom an anomalous coronary vessel was found at a single center. Metoprolol (50-100 mg) was given orally to all patients to reduce heart rate so as to get high-quality MDCT images. After performing MDCT, the CT scans of each patient were analysed and compared with their coronary angiograms by two experienced radiologists and one cardiologist who were unaware about the study, and the sensitivity of MDCT was determined. RESULTS: Twenty-three patients (age range 28-73) with seven different coronary arteries of the anomalous type were evaluated. Nineteen patients had an anomalous left coronary artery; three patients had an anomalous single coronary artery; and one patient had an anomalous right coronary artery. The most common anomaly type was the left circumflex coronary artery (52%). The origin and course of all anomalous vessels were detected by ECG-gated MDCT (Lightspeed 16, GE Medical Systems, Milwaukee, Wisconsin, USA). The sensitivity of 100% of MDCT was detected in patients who had anomalous coronary vessels. CONCLUSION: We suggest that MDCT could be a non-invasive alternative imaging technique to conventional coronary angiography for screening the anomalous vessels of coronary arteries because of its excellent spatial resolution, which is very important for detecting the relationship of anomalous vessels with great arteries and cardiac structures. PMID- 18418239 TI - The efficacy of adjunctive balloon postdilation at the overlapping site of drug eluting stent in diffuse long coronary lesion. AB - BACKGROUND: Data regarding the efficacy of adjunctive balloon postdilation (ABP) at the site of two or more overlapping drug-eluting stent (DES) implantation in diffuse long lesion angioplasty are limited. This study was aimed to evaluate the efficacy of routine ABP to the overlapping DES sites in terms of clinical and angiographic outcomes. METHODS: We enrolled 88 patients (55 men, mean age 63.51+/ 10.24 years), and 93 lesions with diffuse long lesion were treated with DES. The clinical and angiographic outcomes up to 6 months of 35 patients in the without routine ABP group were compared with those of 53 patients in the with ABP group. RESULTS: Baseline clinical characteristics, procedural characteristics, angiographic lesion length, lesion type and DES type were similar between the two groups. Late loss and binary restenosis at 6 months were similar between the two groups. The death, Q-wave myocardial infarction and the major adverse cardiac events were similar between the two groups (7.9 vs. 9.1% in ABP group, P=NS), but there was a trend toward lesser target lesion and vessel revascularization in no ABP group (2.6 vs. 9.0% in no ABP group, P=0.09). CONCLUSION: The routine ABP at the overlapping site of DES implantations in a diffuse long coronary lesion showed no significant benefits in terms of angiographic and clinical outcomes compared with those of patients without routine ABP. PMID- 18418240 TI - Bibliography--editors' selection of current world literature. PMID- 18418241 TI - Incoherence with studies using actigraph mti among children age 6-12 yr. PMID- 18418243 TI - Residual functional connectivity in the split-brain revealed with resting-state functional MRI. AB - Split-brain patients present a unique opportunity to address controversies regarding subcortical contributions to interhemispheric coordination. We characterized residual functional connectivity in a complete commissurotomy patient by examining patterns of low-frequency BOLD functional MRI signal. Using independent components analysis and region-of-interest-based functional connectivity analyses, we demonstrate bilateral resting state networks in a patient lacking all major cerebral commissures. Compared with a control group, the patient's interhemispheric correlation scores fell within the normal range for two out of three regions examined. Thus, we provide evidence for bilateral resting state networks in a patient with complete commissurotomy. Such continued interhemispheric interaction suggests that, at least in part, cortical networks in the brain can be coordinated by subcortical mechanisms. PMID- 18418244 TI - Music in minor activates limbic structures: a relationship with dissonance? AB - Using functional magnetic resonance imaging, we contrasted major and minor mode melodies controlled for liking to study the neural basis of musical mode perception. To examine the influence of the larger dissonance in minor melodies on neural activation differences, we further introduced a strongly dissonant stimulus, in the form of a chromatic scale. Minor mode melodies were evaluated as sadder than major melodies, and in comparison they caused increased activity in limbic structures, namely left parahippocampal gyrus, bilateral ventral anterior cingulate, and in left medial prefrontal cortex. Dissonance explained some, but not all, of the heightened activity in the limbic structures when listening to minor mode music. PMID- 18418245 TI - A high-resolution large area serotonin map of a live rat brain section. AB - We employ three-photon microscopy to produce a high-resolution map of serotonin autofluorescence in a rat midbrain section (covering more than half of the brain), to quantitatively characterize serotonin distribution and release in different areas of a live brain slice. The map consists of a tiling of approximately 160 contiguous optical images (covering an area of approximately 27 mm with sub-mum resolution in 20 min), and is recorded before and after inducing depolarization. We observe that the total serotonin exocytosed from the somata in the raphe is quantitatively comparable with regions containing a high density of serotonergic processes. Our results demonstrate that high-resolution, wide-area, dynamic neurotransmitter mapping is now possible. PMID- 18418246 TI - Different dorsal horn neurons responding to histamine and allergic itch stimuli. AB - We examined whether different itch signals converge on the same dorsal horn neurons in mice. Intradermal injections of histamine and SLIGRL-NH2 (protease activated receptor-2 agonist) induced scratching in naive mice and so did mosquito allergen in sensitized mice. These stimuli induced Fos expression in cells in the superficial dorsal horn. Fos-positive cells were mainly distributed within the isolectin B4-labeled region (inner aspect of lamina II) after histamine injection. In contrast, they were in the region dorsal to the isolectin B4-labeled region after injections of SLIGRL-NH2 and mosquito allergen. These results suggest that allergic itch signal is mediated by primary afferents expressing protease-activated receptor-2 and the neurons receiving signals of protease-associated itch and allergy-associated itch are different from those of histamine-induced itch. PMID- 18418247 TI - Changes in corticospinal excitability during observation of walking in humans. AB - To address whether the passive observation of walking would induce an increase in motor cortical excitability, we examined the responses of motor-evoked potential elicited by transcranial magnetic stimulation in the tibialis anterior and soleus muscles as the participants observed naturally performed walking. Motor-evoked potentials in these muscles were significantly increased during the observation of walking throughout the entire step-cycle periods, but not during specific step periods. These findings indicate that cortical excitability can be increased not only during the observation of voluntary hand/arm movements, but also during the observation of automatic movements such as walking. It is also suggested that the present results may reflect the increased cortical excitability during the entire walking cycle. PMID- 18418248 TI - N-methyl-D-aspartate-dependent long-term potentiation of excitatory transmission in trigeminal subnucleus oralis. AB - This study for the first time demonstrates early developmental changes of passive/active membrane properties, and long-term potentiation (LTP) of excitatory synaptic transmission at spinal trigeminal subnucleus caudalis (Vc)-to oralis (Vo) synapses. During postnatal development, the probability of Vo neurons with monosynaptic excitatory postsynaptic currents (EPSCs) upon Vc stimulation significantly increased, whereas the input resistances of Vo neurons and the latencies of monosynaptic EPSCs significantly decreased. Application of a 'pairing' protocol that comprises 2 Hz-conditioning stimulation of Vc with postsynaptic depolarization of Vo neuron to +30 mV generated LTP of alpha-amino-3 hydroxy-5-methylisoxazole-4-propionic acid receptor-mediated monosynaptic EPSC amplitude in more than 70% of Vo neurons. The induction of LTP required the activation of N-methyl-D-aspartate receptor, but its magnitudes had correlation neither with postnatal ages nor with baseline EPSC amplitudes. PMID- 18418249 TI - Novel monoamine oxidase A knock out mice with human-like spontaneous mutation. AB - A novel line of mutant mice [monoamine oxidase A knockout (MAOA KO)] harboring a spontaneous point nonsense mutation in exon 8 of the MAO A gene was serendipitously identified in a 129/SvEvTac colony. This mutation is analogous to the cause of a rare human disorder, Brunner syndrome, characterized by complete MAO A deficiency and impulsive aggressiveness. Concurrent with previous studies of MAO A KO mice generated by insertional mutagenesis ('Tg8'), MAOA(A863T) KO lack MAO A enzyme activity and display enhanced aggression toward intruder mice. MAOA(A863T) KO, however, exhibited lower locomotor activity in a novel, inescapable open field and similar immobility during tail suspension compared with wild type, observations which differ from reports of Tg8. These findings consolidate evidence linking MAO A to aggression and highlight subtle yet distinctive phenotypical characteristics. PMID- 18418250 TI - When Chinese semantics meets failed syntax. AB - Previous event-related potential studies in Indo-European languages reported a surprising finding that failed syntactic category processing appears to block lexical-semantic integration, suggesting a functional primacy of syntax over semantics. An event-related potential experiment was conducted to test whether there is such primacy in Chinese sentence reading, using sentences containing either semantic only violations, combined syntactic category and semantic violations, or no violations. Semantic only violations elicited a centro-parietal negativity and combined violations a broadly distributed, but centro-parietally focused negativity, both in the 300-500 ms window and followed by a P600, suggesting that semantic integration proceeds even when syntactic category processing fails. Thus, there is no functional primacy of syntactic category over semantic processes during Chinese sentence reading. PMID- 18418251 TI - Nrxn3 upregulation in the globus pallidus of mice developing cocaine addiction. AB - Dysfunctions affecting the connections of basal ganglia lead to major neurological and psychiatric disorders. We investigated levels of mRNA for three neurexins (Nrxn) and three neuroligins (Nlgn) in the globus pallidus, subthalamic nucleus, and substantia nigra, in control conditions and after short-term exposure to cocaine. The expression of Nrxn2beta and Nlgn3 in the substantia nigra and Nlgn1 in the subthalamic nucleus depended on genetic background. The development of short-term cocaine appetence induced an increase in Nrxn3beta expression in the globus pallidus. Human NRXN3 has recently been linked to several addictions. Thus, NRXN3 adhesion molecules may play an important role in the synaptic plasticity of neurons involved in the indirect pathways of basal ganglia, in which they regulate reward-related learning. PMID- 18418252 TI - Thrombin regulates CD40 expression in microglial cells. AB - Microglial cells are the innate immune cells of the central nervous system and quickly respond to injury by proliferation, cytokine release, and increased cell surface antigen expression. Thrombin is a multifunctional serine proteinase, which has the capability to activate microglial cells. Here, we report that pharmaceutical-grade thrombin dose-dependently increases the expression of CD40 in N9 microglial cells. This effect is blocked by a thrombin inhibitor, mimicked by thrombin receptor-activating peptide and modified by mitogen-activated protein kinase pathway inhibitors. Thrombin-induced CD40 regulation might play a role in diseases with breakdown of the blood-brain barrier such as multiple sclerosis or stroke. PMID- 18418253 TI - Identity versus similarity priming for letters in left mid-fusiform cortex. AB - Using functional magnetic resonance imaging, this study examined neural responses in the Visual Word Form Area to prime - test letter pairs in which visual similarity and identity were manipulated. Results revealed the greatest priming for pairs with high visual similarity, less priming for pairs with medium similarity, and the least priming for pairs with low similarity. Moreover, when visual similarity was equated, priming magnitude did not differ for pairs with the same letter identity compared with those with different letter identities. As such, results contrast with views of the Visual Word Form Area as supporting arbitrary (nonsimilarity based) mapping requirements of reading, and suggest a less modular perspective on visual form recognition. PMID- 18418254 TI - Hepatocyte growth factor promotes proliferation and migration in immortalized progenitor cells. AB - Hepatocyte growth factor (HGF) and its receptor c-Met are widely expressed in the developing and adult brain. However, little is known about the role of HGF during the development of the human dopaminergic neuronal system. We have established telomerase-immortalized dopaminergic progenitor cells isolated from the fetal striatum that express markers for neural progenitor cells and tyrosine hydroxylase. We show that the cells were able to differentiate into dopaminergic neurons and release dopamine. Exogenous HGF-induced proliferation was inhibited by U0126, whereas migration was completely blocked by LY294002. Study demonstrates that HGF regulates the proliferation and migration of dopaminergic progenitor cells. Modulating dopaminergic progenitor cells in the striatum may prove to be a new approach for treating Parkinson's disease. PMID- 18418255 TI - Resynchronization in neuronal network divided by femtosecond laser processing. AB - We demonstrated scission of a living neuronal network on multielectrode arrays (MEAs) using a focused femtosecond laser and evaluated the resynchronization of spontaneous electrical activity within the network. By an irradiation of femtosecond laser into hippocampal neurons cultured on a multielectrode array dish, neurites were cut at the focal point. After the irradiation, synchronization of neuronal activity within the network drastically decreased over the divided area, indicating diminished functional connections between neurons. Cross-correlation analysis revealed that spontaneous activity between the divided areas gradually resynchronized within 10 days. These findings indicate that hippocampal neurons have the potential to regenerate functional connections and to reconstruct a network by self-assembly. PMID- 18418256 TI - Chronometric features of processing unpleasant stimuli: a functional MRI-based transcranial magnetic stimulation study. AB - The quick identification of potentially threatening events is a crucial cognitive capacity to survive in a changing environment. Previous functional MRI data revealed the right dorsolateral prefrontal cortex and the region of the left intraparietal sulcus (IPS) to be involved in the perception of emotionally negative stimuli. For assessing chronometric aspects of emotion processing, we applied transcranial magnetic stimulation above these areas at different times after negative and neutral picture presentation. An interference with emotion processing was found with transcranial magnetic stimulation above the dorsolateral prefrontal cortex 200-300 ms and above the left intraparietal sulcus 240/260 ms after negative stimuli. The data suggest a parallel and conjoint involvement of prefrontal and parietal areas for the identification of emotionally negative stimuli. PMID- 18418257 TI - The effects of pitch and pitch strength on an auditory-evoked N1m. AB - The aim of this paper was to determine whether the latency and/or amplitude of the N1m deflection of the auditory-evoked magnetic fields are influenced by the delay and number of iterations of iterated rippled noise, which are related to pitch and pitch strength, respectively. The results indicate that the N1m amplitude decreased sharply for delays between 16 and 32 ms, suggesting that the N1m amplitude reflects the lower limit of the audible pitch range. The N1m latency increases with increasing delay of up to 8-16 ms and then decreases again for delays longer than 16 ms. The behavior of the latency may reflect the balance between the pitch-related component of the N1m and a specific pitch-unrelated component. PMID- 18418258 TI - Conjugated equine estrogen enhances rats' cognitive, anxiety, and social behavior. AB - The ovarian hormone, 17beta-estradiol (E2), has numerous targets in the body and brain, and can influence cognitive, affective, and social behavior. However, functional effects of commonly prescribed E2-based hormone therapies are less known. The effects of conjugated equine estrogen (CEE) on middle-aged female rats for cognitive (object recognition), anxiety (open field, plus maze), and social (social interaction, lordosis) behavior were compared with vehicle. Our hypothesis that CEE would enhance cognitive, anxiety, and social behavior was supported. CEE improved object recognition, increased time spent on the open arms of the plus maze, and time spent interacting with a conspecific, but did not alter open field behavior or lordosis. Thus, CEE can enhance cognitive, antianxiety, and social behavior of middle-aged rats. PMID- 18418259 TI - Cervical spinal cord injury in abused children. AB - Five infants and toddlers who sustained cervical spinal cord injury as the result of child abuse are described. Three cases are previously unreported. Diagnosis was complicated by coexistent brain injuries and their treatments, subtle and/or evolving paralysis, and central cord syndrome, in which arm function is diminished but leg function is preserved. Definitive spinal imaging by magnetic resonance imaging (MRI), computed tomography, and plain radiographs was delayed because of life support efforts. When completed, the MRI was most sensitive to cord injury. Evidence of associated bony spinal injury was often absent or unapparent until healing occurred; 4 children had spinal cord injury without (or with minimal) radiological abnormality. The 3 children presenting to our hospital with cord injury represent 1% of the estimated cases of inflicted head injury seen during a 23-year period. PMID- 18418260 TI - Curanderismo: consequences of folk medicine. AB - Curanderismo, folk medicine, is an important and common aspect of Hispanic culture. Its use is not well understood by US medical physicians and is often overlooked when Hispanic patients present to US hospitals. We present a case of isopropyl alcohol toxicity in a 4-year-old child due to the use of a curanderismo treatment of "espanto" (evil spirits). PMID- 18418261 TI - An uncommon presentation of stroke in a child with trisomy 21. AB - A 3-year-old boy was presented to the emergency department with fever and refused to bear weight on his left leg. Evaluation leads to the eventual diagnosis of stroke secondary to moyamoya syndrome. This is an unusual presentation of stroke and highlights the need to expand the differential diagnosis of common presentations to include rare diseases in children with predisposing conditions. We explore the relationship between trisomy 21 and moyamoya syndrome and then briefly discuss strokes in childhood. PMID- 18418262 TI - Systemic allergic reaction to tree processionary caterpillar in children. AB - Contact with the tree processionary caterpillar or ingestion/inhalation of its airborne toxin-containing hairs and spines may cause a direct histamine-releasing or IgE-mediated allergic reaction in humans. The most common manifestations are urticaria and dermatitis; systemic manifestations such as respiratory involvement and anaphylaxis have been reported mainly in foresters. We describe a 12-year-old child who was presented to the emergency department with systemic allergic reaction (pruritic rash, conjunctival hyperemia, dyspnea, and wheezing), which eventually proved to be due to exposure to the pine tree processionary caterpillar. The diagnosis was made only after the child's repeated exposure to the caterpillar. Tree processionary caterpillar should be added to the list of causes of systemic allergic reactions in children, especially in endemic areas, and medical staff should be alerted the range of clinical manifestations to ensure prompt diagnosis and treatment. PMID- 18418263 TI - Traumatic tracheal laceration in a pediatric patient medically managed with high frequency oscillatory ventilation. AB - A 12-year-old boy presented to the emergency department in respiratory failure with massive subcutaneous emphysema extending from his head to his thighs. This resulted after he fell onto an industrial trash receptacle (dumpster) striking his right lateral neck. Airway management was obtained through an oral endotracheal tube in the emergency department. Initial scans revealed massive subcutaneous emphysema and pockets of anterior air suggestive of a tracheal tear. The patient was conservatively managed without surgical repair using high frequency oscillatory ventilation through an oral endotracheal tube. Injuries to the trachea are exceedingly rare in the pediatric population. These patients have a high morbidity and mortality likely due to delay in proper diagnosis of this injury. Historically, these patients have been treated with surgical correction of the injury. Management of tracheal tears without surgical intervention may be possible if the endotracheal tube is able to stent the wound and allow for wound healing. PMID- 18418264 TI - Acute suppurative sialadenitis in a 37-day-old girl. AB - Sialadenitis is a rare diagnosis in early infancy, but it must be considered in the differential diagnosis of neck swelling in this age group. We present a case of a 37-day-old infant with sialadenitis who presented to the pediatric emergency department with fever and neck swelling. A discussion of this diagnosis and the relevant literature follows. PMID- 18418265 TI - Pediatric emergency medicine: legal briefs. PMID- 18418266 TI - A practical approach to warts in the emergency department. AB - Although cutaneous warts are one of the most common skin conditions affecting children, their management can be challenging, especially in complex cases and even more so during a single emergency department encounter. Anogenital warts require particular attention because of their possible but nonspecific association with sexual abuse. This review will focus on cutaneous warts and anogenital warts, with a discussion of their recognition and treatment, especially in the emergency department. The medicolegal implications of anogenital warts in children are also reviewed. PMID- 18418268 TI - Outcomes evaluation of the 2005 National Pediatric Emergency Medicine Fellows' Conference. AB - OBJECTIVE: Evaluate the effectiveness of the 2005 National Pediatric Emergency Medicine Fellows' Conference in achieving predefined objectives in scholarship, leadership, and partnership. METHODS: Preconference and postconference questionnaires measured knowledge, research-related confidence, beliefs about institutional support for emergency medical services for children (EMSC) research, and intentions to engage in 7 specific behaviors relating to scholarship, leadership, and partnership. Bivariate correlations among the variables were assessed. Repeated-measures analysis of variance measured change between preconference and postconference measures. Hierarchical multiple linear regression models identified predictors of postconference intentions to engage in each of the 7 specific behaviors controlling for preconference intentions. RESULTS: Ninety-two pediatric emergency medicine fellows attended; more than 90% completed preconference and postconference questionnaires, and 88% completed both. Knowledge (P < 0.001) and confidence (P < 0.001) improved after the conference. Increases were observed in fellows' intentions to establish national mentoring relationships (P < 0.001), continue in EMSC research (P = 0.003), and participate in national collaborations (P = 0.022). Confidence and subjective norms correlated significantly (P < 0.05) with each of the behavioral intentions in bivariate tests. In the multivariable regression models, confidence correlated with 4 behavioral intentions (P < 0.02), and subjective norms correlated with one (P < 0.001). Forty-eight percent established a mentoring relationship with a faculty member; 25% planned to collaborate on research with other attendees. CONCLUSIONS: The 2005 National Pediatric Emergency Medicine Fellows' Conference resulted in positive, measurable outcomes relating to the professional development of attendees, including increased conference-specific knowledge, confidence, and intentions to continue in EMSC research, join national collaborative research networks, and establish national mentoring relationships. PMID- 18418269 TI - Rhabdomyolysis: a review of clinical presentation, etiology, diagnosis, and management. AB - Rhabdomyolysis is a condition that results from many underlying etiologies and can present in a myriad of ways to the emergency physician. However, some clinical and laboratory features are almost always present and, if noted, can help in making the diagnosis. This review article will focus on the presenting symptoms, the various etiologies, the underlying mechanisms, and the current management of pediatric rhabdomyolysis. PMID- 18418270 TI - ECGs in the ED. PMID- 18418272 TI - Antiplatelet therapy in early management of non-ST-segment elevation acute coronary syndrome: the 2002 and 2007 guidelines from North America and Europe. AB - The American College of Cardiology, American Heart Association, and the European Society of Cardiology published updated guidelines in 2007 for patients with non ST elevation acute coronary syndrome. In this article, we review the recommendations for antiplatelet therapy and supporting data, highlight new changes, and describe differences between the European and North American guidelines. The new guidelines provide more details regarding the selection of an early conservative versus an early invasive approach based on the patient's profile and balance between ischemic and bleeding risks. Important new recommendations include wider endorsement for low-dose aspirin maintenance therapy, longer duration of clopidogrel following percutaneous coronary intervention, additional guidance regarding surgery in selected patients on clopidogrel, identification of patients most likely to benefit from glycoprotein IIb/IIIa inhibitors (with appropriate dose modification in patients with renal failure), and the option to use early clopidogrel with bivalirudin in patients managed invasively who are at increased risk of bleeding. The new guidelines also discourage the concomitant use of nonsteroidal anti-inflammatory drugs and delineate indications for adding warfarin to antiplatelet therapy. Because antiplatelet therapy is the cornerstone of management of patients with non-ST elevation acute coronary syndrome, health care providers should make themselves familiar with the new data and latest guideline recommendations. PMID- 18418273 TI - Prevention of salt-induced hypertension and fibrosis by AT1-receptor blockers in Dahl S rats. AB - In Dahl S rats, high salt intake causes hypertension and cardiovascular hypertrophy and fibrosis, associated with an apparent increase in activity of tissue RAAS. In the current study, we assessed the effects of two AT1-receptor blockers (ARB) on AT1- and AT2-receptors and ACE densities and salt-induced cardiovascular changes. The hydrophilic ARB losartan (30 or 100 mg.kg.d) and the lipophilic ARB telmisartan (10 or 30 mg.kg.d) were administered once daily, and a high-salt diet was provided from 5 to 9 weeks. In Dahl S but not R rats, the high salt diet caused marked hypertension, cardiac and kidney hypertrophy, and fibrosis. Both ARBs dose-dependently inhibited binding of Ang II to AT1-receptors and reversed the salt-induced increases in AT2-receptor densities in the CNS. Both ARBs at regular doses attenuated the salt-induced hypertension and, at high doses, prevented the increase in BP during the day but not during the night. Both ARBs similarly prevented high-salt-induced interstitial and perivascular fibrosis in the LV and RV as well as fibrosis in the aorta and renal tubules. RV hypertrophy was also prevented, but LV hypertrophy only partially, and kidney hypertrophy not at all. In Dahl S rats, AT1-receptor stimulation seems to play a critical role in salt-induced hypertension and fibrosis, but a lesser role in tissue hypertrophy. PMID- 18418275 TI - Tumor volume as a prognostic factor in oropharyngeal squamous cell carcinoma treated with primary radiotherapy. AB - OBJECTIVES/HYPOTHESIS: Tumor volume has been demonstrated to play a prognostic role in many head and neck cancers. The purpose of this study was to conduct an institutional review analyzing the correlation between tumor volume and locoregional control of oropharyngeal squamous cell cancer treated with primary radiotherapy. STUDY DESIGN: Retrospective institutional chart analysis. METHODS: Seventy-nine patients from 1991 to 2005 with primary T1 to T4 oropharyngeal squamous cell carcinoma (base of tongue, n = 31; soft palate, n = 1; tonsils, n = 47) were treated with primary radiotherapy. Tumor volumes were measured from pretreatment computerized tomography scans by two observers. Three-dimensional tumor volumes were calculated using a computer digitizer for each computed tomography slice showing the primary lesion. Survival analysis, using the methods of Kaplan and Meier, was performed to assess whether tumor volume, Tumor, Node, Metastasis classification, tumor stage, or location were associated with locoregional failure. RESULTS: Tumor volume did not significantly correlate with locoregional failure (observer 1, P = .6244; observer 2, P = .5612). There was a high interobserver correlation (r = 0.98970). Univariate analysis did, however, demonstrate a significant difference in locoregional failure between T4 tumors and all other T stages (T1 vs. T4, P = .0107; T2 vs. T4, P = .0004; T3 vs. T4, P = .0155). Nodal status, tumor stage, and location did not significantly correlate with locoregional failure rate. CONCLUSIONS: Tumor volume does not appear to play a significant role in predicting locoregional recurrence for patients with primary squamous cell cancer of the oropharynx treated with primary radiotherapy. However, T4 status was predictive of poor locoregional control. PMID- 18418274 TI - Role of protein kinase G in nitric oxide deficiency-induced supersensitivity to nitrovasodilator in rat pulmonary artery. AB - The aim of the present study was to examine the role of protein kinase G (G kinase) in the mechanism of endogenous nitric oxide (NO) deficiency-induced supersensitivity to the nitrovasodilator sodium nitroprusside (SNP) in isolated rat pulmonary artery. Tension experiments and cGMP measurements were carried out on isolated rat pulmonary artery to assess the influence of NO deficiency, caused by either N-nitro-L-arginine methyl ester (L-NAME) treatment or endothelium removal on the vasodilator potency of SNP. Sodium nitroprusside was more potent (pD2; 8.21 +/- 0.04) in relaxing arterial rings treated with 100microM L-NAME or denuded of the endothelium (pD2; 8.44 +/- 0.11) compared with the endothelium intact controls (pD2; 7.61 +/- 0.05). Similarly, the tissue sensitivity to 8-Br cGMP, a G-kinase activator, was significantly (P < 0.05) greater after L-NAME treatment (pD2; 5.04 +/- 0.09) or endothelium removal (pD2; 5.28 +/- 0.11) in comparison with the controls (pD2; 4.22 +/- 0.17). On the other hand, dibutyryl cAMP, an activator of protein kinase A, was equipotent in dilating control (pD2; 4.14 +/- 0.04) and L-NAME-treated (pD2 4.21 +/- 0.05) vessels. Further, L-NAME treatment significantly (P < 0.05) decreased the basal cGMP but enhanced SNP (1 microM)-stimulated increase in the tissue cyclic nucleotide levels (271.8 +/- 39.93 pmol/mg protein versus control: 66.19 +/- 7.18 pmol/mg protein), indicating sensitization of soluble guanylyl cyclase to NO. The increased sensitivity of G kinase to cGMP observed in the present study suggests a novel mechanism of supersensitivity in vascular smooth muscle to nitrovasodilators in acute NO deficiency. Further, it explains the influence of ambient cGMP in determining the sensitivity of G-kinase in vascular smooth muscle. PMID- 18418276 TI - Lingual thyroid in children: a rare clinical entity. AB - OBJECTIVES/HYPOTHESIS: To study the presentation, management, and long-term outcome of children presenting with lingual thyroid. STUDY DESIGN: Institutional review board approved, retrospective study (1993-2004). METHODS: The study was conducted at a tertiary care pediatric medical center. The main outcomes measured were initial presentation, radiographic findings, endocrine evaluation, surgical outcome, pathologic features, complications, need for hormonal replacement. RESULTS: Four patients presented to the Department of Otolaryngology and Communications Enhancement, Children's Hospital Boston with lingual thyroid between 1993 and 2004. All patients were female, with an age range of 2 to 12 years (x = 6). All patients presented with a mass (1.4-3.5 cm) and most with respiratory or feeding difficulty. Magnetic resonance imaging was obtained in three patients and revealed a mass consistent with lingual thyroid. Thyroid scan confirmed the lingual thyroid as the only functioning thyroid in all four patients. None of the patients responded to hormonal replacement, and all underwent surgical excision of the mass. Surgical approach included midline glossotomy (n = 2) and CO2 laser excision (n = 2). Pathologic evaluation confirmed lingual thyroid in all four patients. No evidence of malignancy was seen in any patient. All four patients require lifelong hormonal replacement. CONCLUSIONS: Lingual thyroid is a rare condition, with an incidence of 1:100,000. This infrequent congenital anomaly is often asymptomatic until a pathologic stress such as systemic disease or physiologic stress such as puberty causes enlargement of the ectopic tissue, leading to dysphagia, dysphonia, and dyspnea. The work-up should include routine blood work including thyroid function tests thyrotropin, thyroxine, and thyroid hormone binding ratio; iodine thyroid scintigraphy; and computerized tomography or magnetic resonance imaging. The majority of patients require surgical excision of the symptomatic mass and, in case of absence of orthotopic thyroid tissue, long-term thyroid hormone replacement. PMID- 18418277 TI - Failed tube extrusion is not a random event in children or their siblings. AB - OBJECTIVES/HYPOTHESIS: It has been assumed that prolonged retention of tympanostomy tubes was random, with an incidence of 4% to 12% per tube. We have noticed patterns of tube retention that suggested a genetic predilection. We undertook this study to validate or disprove this impression. Such information may shed light on the physiology of tube extrusion and may help better define the risks of tympanostomy tube placement. STUDY DESIGN: Retrospective, single surgeon, statistical analysis. METHODS: A computerized database of approximately 10,000 pediatric otolaryngology patients was queried. We identified all children treated by the senior author during a 6-year period who underwent tube removal for failed extrusion 24 or more months after placement. Siblings with retained tubes and individuals requiring bilateral tube removal were further analyzed: 1) using the law of multiplication of independent factors, the chance of retaining tubes in both ears of an individual was compared with the observed rate of bilateral retention by calculating the difference in Poisson rates; 2) the rate of siblings who retained one or both tubes was compared with nonsiblings who retained one or both tubes using the Z test for independent proportions. RESULTS: The observed rate of bilateral retention (228/6,000 = 0.038) far exceeded the theoretical rate (0.068 = 0.0046; P < .001). Siblings were more likely to retain tubes than nonsiblings (difference = 0.042; P = .085). This difference was more pronounced in unilateral (difference = 0.048; P = .014) than in bilateral retention (difference = 0.005; P = .856). CONCLUSIONS: Tympanostomy tube retention is not a random occurrence. Patterns of nonextrusion in individuals and siblings suggest a genetic mechanism. Implications for patient counseling and research are discussed. PMID- 18418278 TI - Prognostic impact of primary tumor volume in patients with nasopharyngeal carcinoma treated by definitive radiation therapy. AB - OBJECTIVES/HYPOTHESIS: Tumor burden has been confirmed as one of the important indicators in disease control after treatment for various types of malignancies. This report aims to document the value of the primary tumor volume of nasopharyngeal carcinoma [gross tumor volume of the primary site (GTV-P)] in predicting the treatment outcome after high-dose definitive radiation therapy. STUDY DESIGN: A total of 154 patients with nasopharyngeal carcinoma were prospectively treated with accelerated hyperfractionated radiotherapy alone to a total dose of 78 Gy/60 fractions/6 weeks (biological effective dose = 88.2 Gy). METHODS: Patients were assessed for local control, disease-free survival and distant failure free survival. The GTV-P was calculated by summation of area technique after delineation of the primary tumor contours on computed tomography images. RESULTS: The median GTV-P was 14.5 mL for T1, 23.3 mL for T2, 35.8 mL for T3, and 50.9 mL for T4 diseases (P < .05). With a median follow-up of 61 months, the 5-year local failure-free rate, disease-free survival and distant failure free survival rates were 89.4% versus 48.9% (P = .002), 56.6% versus 0% (P = .001), and 66.9% versus 16.5% (P = .0001), respectively, for patients whose GTV-P were < or =60 mL and >60 mL. Multivariate analysis revealed that GTV-P is an independent prognostic factorfor local control (hazard ratio = 3.568, P = .035). There fore, the GTV-P was significantly associated with the treatment outcome after high-dose radiation delivered in accelerated hyperfractionated schedule for nasopharyngeal cancer patients. CONCLUSIONS: Tumor volume is a reliable indicator for supplementing the T classification of the Tumor, Node, Metastasis staging system for predicting local control after definite radiation therapy. PMID- 18418279 TI - Medical management of Meniere's disease. AB - OBJECTIVES: Meniere's disease (MD) is poorly understood with respect to its etiology, pathophysiology, clinical course, and treatment. Furthermore, in the absence of controlled clinical studies, empiric treatments have been used with varying degrees of success. In this paper, the authors review the current medical management of MD. STUDY DESIGN: Literature review. RESULTS: Because of a dearth of well-controlled studies, the medical management of MD remains empirical and is largely restricted to lifestyle changes, pharmacotherapy, and office-based procedures. The development of transtympanic therapies represents a true therapeutic advance that has largely supplanted surgical intervention. A treatment algorithm for acute and chronic medical management is reviewed. CONCLUSIONS: Despite absence of a complete understanding of MD, medical management or its natural history leads to control of vertigo in the majority of patients. Basic research is needed to understand its pathophysiology so that directed therapies can be developed and can be tested in well-controlled clinical trials. PMID- 18418280 TI - Sleep medicine for surgeons. AB - Sleep medicine is an important component of current head and neck surgery practices. Furthermore, obstructive sleep apnea is an anatomic disease of the upper respiratory tract, a region of anatomy best known to head and neck surgeons. If head and neck surgeons choose to participate in the surgical treatment of sleep apnea, they must develop expertise in the evaluation and home sleep testing aspects of sleep apnea. Because positive airway pressure remains the front line treatment, they must also develop expertise in the prescription of positive airway pressure therapy. PMID- 18418281 TI - Asymmetric hearing loss: definition, validation, and prevalence. AB - HYPOTHESIS: An algorithm for identifying asymmetric hearing loss (AHL) can be constructed that performs as well or better than expert judges. BACKGROUND: AMCLASS is a method for classifying audiograms based on configuration, severity, site of lesion, and interaural asymmetry. The development and clinician validation for all but asymmetry were reported separately. In this report, an algorithm for identifying AHL is described. Using the clinician-validated algorithm, the prevalence of AHL in a database from an academic health center audiology clinic was analyzed. METHODS: : Five expert clinicians classified 199 audiograms as symmetric or asymmetric. Interjudge agreement was analyzed for each pair of judges and between each judge and the consensus of the panel. An algorithm was constructed based on the set of rules that maximized agreement between AMCLASS and judges. Using the clinician-validated algorithm, the prevalence of AHL was analyzed for groups based on quantity of bone conduction testing, hearing loss configuration, severity, and site of lesion. RESULTS: There was substantial disagreement among judges that was similar to interjudge comparisons for other medical tests. Average agreement between AMCLASS and the judges was higher than agreement between the best judge and the consensus of the judges. Approximately 50% of all patients and 55% of patients with sensorineural hearing loss were classified as AHL by the clinician-validated algorithm. CONCLUSION: The algorithm met the goal of equaling or exceeding the performance of expert judges. The prevalence of AHL was higher than expected and suggests that the algorithm is not useful for screening for acoustic neuroma or other conditions. Perhaps, a criterion based on the magnitude of the asymmetry would better serve that purpose. The symmetry category provided by AMCLASS provides a determination of clinically significant AHL that agrees with the consensus of expert judges. PMID- 18418282 TI - Anterior semicircular canal dehiscence and cranial vibration-induced nystagmus test. PMID- 18418283 TI - Serous otitis media in children: implication of Alloiococcus otitidis. AB - OBJECTIVE: Otitis media with effusion (OME) has long been considered to be a noninfective disease resulting from a eustachian tube dysfunction. However, several microbiological techniques have shown bacteria in the middle ear fluids from patients with OME. Alloiococcus otitidis has been detected in the middle ear fluid from patients with OME. The exact role of this infectious agent in the pathogenesis of OME has yet to be elucidated. STUDY DESIGN: A prospective clinical trial. SETTING: A tertiary university-based referral center. PATIENTS: The inclusion criteria included 110 patients aged between 1 and 12 years. The control group included samples obtained from 30 healthy children undergoing a cochlear implantation. The second group included 40 middle ear effusions (MEEs) that were collected from 40 pediatric OME patients during the placement of the ventilation tube. As for the third group, they were 40 children with acute otitis media. INTERVENTION: The bacterial analysis of the MEE was performed by means of microbiological culture-specific techniques. MAIN OUTCOME MEASURE: Positive cultures for A. otitidis as analyzed by bacteriological analysis of samples from the middle ear mucosa and MEE. RESULTS: Bacteria were present in the culture of 72.5% (29) of the patients with OME. Alloiococcus otitidis was the most frequent bacterium in OME (48.27%) as well as Haemophilus influenzae nonserotype B (17.24%). Streptococcus pneumoniae was the most commonly detected pathogen in acute otitis media (37.5%), and then H. influenzae nonserotype B (25%). For most of the OME cases, only A. otitidis bacteria were isolated. CONCLUSION: We observed a high rate of culture positivity for A. otitidis in patients with clinical OME without suppuration. Further studies are needed to confirm whether the association of A. otitidis with OME represents causality. Antibiotic therapy aimed at A. otitidis is complicated by reported resistance, thus emphasizing the importance of our understanding of the pathogenetic role played by this organism PMID- 18418284 TI - A new double-cartilage block ossiculoplasty: long-term results. AB - OBJECTIVES: To determinate whether ossicular chain reconstruction performed with modified double-cartilage block (DCB) ossiculoplasty used by authors, leaving the intact perichondrium layer on the cartilage surface in contact with tympanic membrane, results in significant hearing results and prevents DCB displacement and DCB loss of stiffness when compared with the original DCB technique described by Luetje and Denninghoff. MATERIALS AND METHODS: Sixty-five ossicular reconstructions using DCB ossiculoplasty were performed from 1996 to 2001 in the whole study group. Eight patients had an inadequate follow-up and were excluded from the study. In the first 25 cases, the original DCB ossiculoplasty technique described by Luetje and Denninghoff was performed (Group 1). In the second group, 32 modified DCB ossiculoplasties were performed. The same author performed all cases. The main follow-up for the whole study group was 7 years. Audiometric data were calculated according to the 1995 American Academy of Otolaryngology-Head and Neck Surgery guidelines. MAIN OUTCOME MEASURES: Mean postoperative air-bone gap closure to less than 20 dB, incidence of DCB displacement from tympanic membrane, and incidence of loss of DCB stiffness. RESULTS: Comparison data of hearing results, incidence of DCB slippage, and incidence of DCB loss of stiffness in the group performed with original technique and in the group with modified technique showed statistical significance. Successful rehabilitation (pure-tone average air bone gap <20 dB) of conductive hearing loss was obtained in 48% of cases in Group 1 (original DCB) and in 81.2% of cases in Group 2 (modified technique). In the group with original DCB ossiculoplasty, the displacement of implant and its loss of stiffness occurred more often than in the second group. CONCLUSION: The modified DCB represents an excellent partial ossicular replacement prosthesis. The perichondrium layer left attached on the cartilage surface in contact with tympanic membrane increases the stability of DCB and prevents the displacement and the loss of stiffness of the cartilage. The modified DCB ossiculoplasty is easy to perform. Hearing results are satisfactory and lasting. The cost is null, and the tolerance is excellent. PMID- 18418286 TI - Auditory cortex stimulation for tinnitus suppression. PMID- 18418287 TI - The use of supplementary techniques to increase recall of sex partners in a network-based research study in Rio de Janeiro, Brazil. AB - OBJECTIVE: To evaluate supplementary cueing as a technique to increase recall of sex partners in the year before the interview. GOAL: Recall of partners beyond those freely recalled. STUDY DESIGN: We asked volunteers at a clinic in Brazil to freely recall all regular and casual sex partners in the year before the interview. Then, we used a name generator developed by Brewer et al. in the United States, in which volunteers were prompted with 4 types of cues: location, alphabetic, social role, and network. We calculated different measures to evaluate the technique and analyzed the associations between reporting any additional partner and demographic characteristics. RESULTS: Among volunteers reporting 2 or more sexual partners (n = 590), 41 (7%) recalled 1 or more additional partners by using the supplementary technique, with 105 partners of 2090 (5%) recalled only after using the cues. For volunteers reporting 4 or more sexual partners (n = 193), 34 (18%) recalled 1 or more additional partners by using the supplementary technique, and 98 of 1177 (8%) of their sexual partners were recalled after using the cues. Men were less likely than women to report sex partners after prompting with the social role cues (OR 0.09), and overall the combined techniques were slightly less effective for older individuals (OR 0.95). CONCLUSION: The cue technique can improve sexual partners' recall in cultural contexts different than the United States, mainly for individuals already reporting several partners before the cue. PMID- 18418288 TI - Environmental support and HIV prevention behaviors among female sex workers in China. AB - OBJECTIVE: To examine the influence of environmental support on HIV prevention behaviors (i.e., condom use behaviors, communication and intention, and HIV/STD testing) among female sex workers (FSWs) in China. METHODS: A cross-sectional survey was administered among 310 FSWs recruited from entertainment establishments in Liuzhou City in Southwest China in 2006. Sex workers were asked about their demographic and working characteristics and their HIV prevention behaviors. Environmental support was measured by a scale consisting of 8 items regarding condom availability, managerial, and social support of HIV prevention (Cronbach alpha = 0.73). RESULTS: After adjusting for demographic and working characteristics, condom use self-efficacy and knowledge, environmental support was strongly associated with following HIV prevention behaviors: overall consistent condom use (aOR = 1.7, 95% CI = 1.3, 2.3), consistent condom use in the last 3 sexual acts (aOR = 1.5, 95% CI = 1.1, 2.0), intention to use condoms consistently (aOR = 1.6, 95% CI = 1.1, 2.2), and HIV testing (aOR = 1.6, 95% CI = 1.1, 2.2). CONCLUSION: Environmental support is a significant predictor of HIV prevention behaviors among establishment-based FSWs in China. The data underscore the importance of developing environmental/structural HIV/STD interventions for FSWs. Condoms need to be easily accessible in the establishments. Gatekeepers need to be educated to create a supportive environment for condom use and HIV prevention. PMID- 18418289 TI - HIV, the clustering of sexually transmitted infections, and sex risk among African American women who use drugs. AB - African American women have high rates of most sexually transmitted infections (STIs), including HIV. STIs have been associated with increased HIV transmission risk. METHODS: Two hundred twenty-eight black women who used drugs completed a structured questionnaire in a central Brooklyn, NY-based research center between March 2003 and August 2005. Women were screened for HIV, herpes simplex virus-2, syphilis, gonorrhea, chlamydia, and trichomoniasis. This analysis determined if STIs cluster within individuals and if clustering and sex practices or partnerships differ by HIV status. RESULTS: Thirty-eight (17%) women tested HIV seropositive and STI prevalence was herpes simplex virus-2 (79%), trichomoniasis (37%), chlamydia (11%), and gonorrhea (2%). Few women knew themselves to be infected with STIs other than HIV. Excluding HIV, the mean number of STIs per woman was 1.3 (SD 0.7). HIV-infected women were significantly more likely than uninfected women to have multiple positive screens (1.6 vs. 1.2, P = 0.002). Women reported having both lower and higher risk sex partners. HIV-infected women were 2 times more likely than uninfected woman to report current sex work (P = 0.05), the only difference in sex risk. In a linear regression model, crack cocaine use was uniquely associated with multiple positive STI screens, excluding HIV (P = 0.002). CONCLUSIONS: Several STIs, including HIV, seem to be endemic among black women who use drugs in this community. In addition to the known geographical clustering of HIV and STIs, STIs were also found to cluster at the individual level. Multiply STI infected individuals may unknowingly, but efficiently, contribute to high STI and HIV rates. PMID- 18418290 TI - A prospective study of risk factors for bacterial vaginosis in HIV-1-seronegative African women. AB - BACKGROUND: Bacterial vaginosis (BV) is common and has been associated with increased HIV-1 susceptibility. The objective of this study was to identify risk factors for BV in African women at high risk for acquiring HIV-1. METHODS: We conducted a prospective study among 151 HIV-1-seronegative Kenyan female sex workers. Nonpregnant women were eligible if they did not have symptoms of abnormal vaginal itching or discharge at the time of enrollment. At monthly follow-up, a vaginal examination and laboratory testing for genital tract infections were performed. Multivariate Andersen-Gill proportional hazards analysis was used to identify correlates of BV. RESULTS: Participants completed a median of 378 (interquartile range 350-412) days of follow-up. Compared with women reporting no vaginal washing, those who reported vaginal washing 1 to 14 [adjusted hazard ratio (aHR) 1.29, 95% confidence interval (CI) 0.88-1.89], 15 to 28 (aHR 1.60, 95% CI 0.98-2.61), and >28 times/wk (aHR 2.39, 95% CI 1.35-4.23) were at increased risk of BV. Higher BV incidence was also associated with the use of cloth for intravaginal cleansing (aHR 1.48, 95% CI 1.06-2.08) and with recent unprotected intercourse (aHR 1.75, 95% CI 1.47-2.08). Women using depot medroxyprogesterone acetate contraception were at lower risk for BV (aHR 0.59, 95% CI 0.48-0.73). CONCLUSIONS: Vaginal washing and unprotected intercourse were associated with increased risk of BV. These findings could help to inform the development of novel vaginal health approaches for HIV-1 risk reduction in women. PMID- 18418291 TI - Acceptability of urine-based screening for Chlamydia trachomatis in asymptomatic young men: a systematic review. AB - We performed a structured literature review of published studies from 2000-2007 that measured the acceptability and acceptance (uptake) of urine testing for C. trachomatis among asymptomatic men. Studies were categorized as three types: (a) non-STD clinic venues where men were approached in person and offered testing on site (urgent care clinics, freestanding clinics or health screening settings, corrections, community centers); (b) delivery of testing kits to men at their homes; and (c) qualitative measurement related to men's experience of screening. When offered in established nonhome-based settings (clinics, schools, corrections), acceptability and consequent uptake of testing by men is generally good (mid-60% range). However, acceptance rates even in these settings vary widely and are influenced by venue, provider, and a diversity of other factors. Acceptance of home-based testing invitations, including direct mailing of test kits, is considerably lower. Attitudinal characteristics of men who decline testing primarily include low self-perception of risk for asymptomatic infection and perceived inconvenience of providing test specimens. Given these findings, testing strategies targeting asymptomatic men in established community and clinic settings are most likely to yield relatively high acceptance rates. However, barriers to both implementation and uptake remain, even when such testing is free. Interventions to enhance uptake of testing in asymptomatic men should be developed and selected with underlying C. trachomatis population prevalence in mind. PMID- 18418292 TI - Patterns of alcohol use, problem drinking, and HIV infection among high-risk African women. AB - OBJECTIVES: To examine the relationship between patterns of alcohol use and HIV infection and to assess the association between problem drinking and the prevalence of risk factors for HIV among a sample of high-risk African women. METHODS: Baseline data were collected between 2002 and 2003 during enrollment of 1050 women in a prospective cohort study designed to assess risk factors for HIV. Information about demographic and employment characteristics, sexual behaviors, and drinking patterns were obtained by interviews. The CAGE questionnaire was used to assess problem drinking. The association between measures of alcohol use and HIV/STDs and sexual behaviors were summarized using odds ratios, adjusted odds ratios (AOR), and 95% confidence intervals (CI). RESULTS: HIV prevalence was 19.0% (95% CI, 16.6%-21.4%). Overall 73.9% of the women drank alcohol whereas 34.6% were classified as problem drinkers. After adjusting for demographic and employment variables, drinkers were at increased risk to be HIV+ when compared with nondrinkers (AOR, 2.10; 95% CI, 1.29-3.42). Greater involvement with alcohol, as indicated by recency, frequency and quantity consumed, was associated with increased risk. Problem drinkers were at greater risk to be HIV+ than nonproblem drinkers (AOR, 1.79; 95% CI, 1.06-3.04 vs. AOR, 2.43; 95% CI, 1.45 4.06). Problem drinkers were also more likely to have engaged in several types of high-risk sexual behaviors and to have other STD infections including HSV-2. CONCLUSION: Programs aiming at limiting alcohol use or promoting abstinence from alcohol might help to reduce high-risk behaviors and lower the burden of HIV/STDs in this population. PMID- 18418293 TI - Vulvovaginal candidiasis in women who have sex with women. AB - OBJECTIVES: To determine whether vulvovaginal candidiasis (VVC) is associated with sexual activity between women. STUDY DESIGN: Cross-sectional survey of 708 new patients attending 2 sexual health clinics for lesbians and bisexual women in London, UK. Questionnaire for demographic variables, sexual history, symptoms, and sexual practice data linked with the results of Gram stain and/or culture of vaginal preparations for identification of Candida species. RESULTS: VVC (either symptomatic or asymptomatic) was common in this sample of women who have sex with women (18.4%). Logistic regression showed that VVC was significantly associated with larger numbers of female sexual partners in the previous year [OR 2.18 (CI 1.35-3.53) for 2 female partners compared with 0 or 1] but not with specific sexual practices, numbers of male partners, use of lubricants or vaginal douching. CONCLUSIONS: The increasing odds of candidiasis with greater numbers of female sexual partners raises the possibility that Candida species could be sexually transmitted between women. PMID- 18418294 TI - Phase I clinical trial of repeat dose terameprocol vaginal ointment in healthy female volunteers. AB - OBJECTIVES: This safety study of terameprocol (also called M4N, EM-1421) daily vaginal application in healthy women explores its potential application as a microbicide in interrupting human immunodeficiency virus sexual transmission and additional interruption of human papillomavirus and herpes simplex virus transmission. METHODS: A double-blind placebo controlled phase I repeat dose tolerability and pharmacokinetic, crossover study of 90 mg terameprocol (2% w/w ointment) administered intravaginally for 7 consecutive days in healthy female subjects. The pharmacokinetics after administration was examined on days 1 and 7 of dosing. Subjects underwent vaginal examination following the 6-hour pharmacokinetic sample on day 7 of each study period. RESULTS: Recruitment started January 2006 and ended May 2006, and 14 subjects completed the study. Median age was 24 years. No treatment-related serious adverse events were reported, and there were a total of 17 treatment-emergent adverse events (AE) reported by 11 participants. The most common AE was headache. Terameprocol was not detectable in serum in pharmacokinetic samples. CONCLUSIONS: Terameprocol was well tolerated at a 90 mg dose (2% wt/wt) administered vaginally daily for 7 days. No serious adverse events occurred and any AEs were mild. The excellent safety profile supports future clinical trial to evaluate the application of intravaginal terameprocol in women. PMID- 18418295 TI - Seroprevalence of herpes simplex virus type 2 and characteristics associated with undiagnosed infection: New York City, 2004. AB - BACKGROUND: Herpes simplex virus type 2 (HSV-2) infection is associated with substantial morbidity and increased risk for human immunodeficiency virus acquisition. We describe HSV-2 seroprevalence in adult New Yorkers, and examine the relationship between select characteristics, infection, and diagnosis. METHODS: HSV-2 seroprevalence and risk factors were measured using the 2004 New York City Health and Nutrition Examination Survey, a population-based cross sectional survey of adults. HSV-2 seroprevalence and corresponding 95% confidence intervals were computed for select characteristics. Associations between proposed risk factors and HSV-2 infection and diagnosis were estimated using unadjusted and adjusted odds ratios. RESULTS: Nearly 28% of adults were infected with HSV-2; 88.4% of HSV-2 positive persons were undiagnosed. Black women had the highest seroprevalence (59.7%) of any sex or race/ethnicity group. Women, non-Hispanic blacks, and Hispanics (vs. non-Hispanic whites), and men who have sex with men were at greater odds of HSV-2 infection. Among HSV-2 infected individuals, non Hispanic blacks (vs. non-Hispanic whites), uncircumcised men, and those with no routine place of care were less likely to be diagnosed. CONCLUSIONS: HSV-2 is highly prevalent and largely undiagnosed in New York City; seroprevalence varies by subgroup. Targeted HSV-2 screening, counseling and treatment may help reduce transmission of HSV-2 and human immunodeficiency virus. PMID- 18418296 TI - Incidence of neonatal herpes simplex virus infections in two managed care organizations: implications for surveillance. AB - OBJECTIVES: To estimate the incidence of neonatal herpes simplex virus (HSV) infections and to assess the utility of surveillance methods for neonatal herpes in 2 managed care populations. METHODS: We identified potential cases using 15 discharge International Classification of Diseases, 9th Revision, Clinical Modification (ICD-9) codes for neonatal HSV and other diseases clinically consistent with this diagnosis. We also searched laboratory databases for positive HSV tests and investigated deaths during the neonatal period. We performed medical chart review using a standardized form. Two pediatric infectious disease specialists reviewed the forms of infants who had a positive HSV test or received a herpes-related diagnosis and made a determination as confirmed, probable, or not a case. RESULTS: Among 270,703 infants born from 1997 to 2002, we identified 737 potential cases and completed medical chart abstraction for 699 (95%). Final review identified 35 confirmed or probable cases of neonatal HSV, and the incidence was 12.9 per 100,000 live births. Only 24 (69%) of the 35 cases were confirmed by laboratory testing. Among the 24 confirmed cases, 22 (92%) received an ICD-9 code of 054.xx or 771.2. Among the 60 infants that received an ICD-9 code of 054.xx or 771.2, only 31 (52%) were confirmed or probable cases of neonatal HSV after final review. CONCLUSIONS: About 30% of neonatal HSV cases were not laboratory confirmed. The use of ICD-9 codes of 054.xx and 771.2 was a sensitive but not specific method to identify cases of neonatal herpes. PMID- 18418297 TI - Health care access and follow-up of chlamydial and gonococcal infections identified in an emergency department. AB - BACKGROUND: We examined 2 potentially important factors influencing successful treatment of Chlamydia trachomatis (Ct) and Neisseria gonorrhoeae (GC) infections identified in an emergency department (ED), health care coverage and reporting the ED as a primary source for health care. METHODS: Adult patients aged 18 to 35 years attending an urban ED were screened for Ct and GC. Patients testing positive were contacted by Disease Intervention Specialists and notified of their infection status. Analyses focus on infected patients for whom we have treatment and follow-up information. We used generalized linear models with log link and binomial error distribution to estimate risk ratios (RRs) and 95% confidence intervals (CI). RESULTS: Of 5537 patients screened in the ED, 348 (6.3%) tested positive for Ct, 143 (2.6%) tested positive for GC, and 43 (0.8%) tested positive for both. Overall, 20% of infected patients did not receive treatment. Among infected patients with no health care coverage 25% (n = 56) were untreated compared with 15% (n = 47) of patients reporting health care coverage (RR: 1.7, 95% CI: 1.2-2.3). Among patients reporting the ED as a primary source for health care 26% (n = 27) were untreated compared with the 18% (n = 77) reporting receiving health care from non-ED sources (RR: 1.4, 95% CI: 1.0-2.1). CONCLUSIONS: EDs often serve as primary care sites for difficult-to-reach populations. We were able to successfully locate and treat the greater part of ED identified infections. However, one-fifth of infected patients did not receive treatment. ED-based screening programs can benefit from integration with local public health infrastructure to improve notification and treatment services. PMID- 18418298 TI - Is jail screening associated with a decrease in Chlamydia positivity among females seeking health services at community clinics?-San francisco, 1997-2004. AB - BACKGROUND: Young adults entering jail are at increased risk for sexually transmitted diseases (STD) such as chlamydia, are released quickly, and are unlikely to be tested for STDs elsewhere. San Francisco jails performed targeted chlamydia screening and treatment since 1996. GOAL: To determine this program's impact on chlamydia positivity among females attending neighborhood medical clinics. STUDY DESIGN: During 1997-2004, jail testing density, a measure of the proportion of persons from year 2000 census blocks that were tested in jail, was compared by neighborhood. Chlamydia positivity among females aged 15 to 25 years were compared at 2 clinics serving areas with different jail testing densities. RESULTS: Of persons offered screening at intake, 89% accepted. A total of 42,952 tests were performed among 23,561 persons in jail (45% black, 73% male). A total of 2765 (6.4%) tests were positive for chlamydia; 81% of chlamydial infections were treated. Jail testing density significantly correlated with neighborhood female chlamydia rates. Mean jail testing density at Clinic S, calculated by using the residence of persons tested for chlamydia, was 7 times greater than that at Clinic O. Chlamydia positivity declined at Clinic S from 16.1% to 7.8% (Ptrend <0.001). No significant change occurred at Clinic O in chlamydia (4.7% in 1997 and 2004, Ptrend = 0.81). CONCLUSIONS: In San Francisco, screening young adults in jail focused testing on persons from neighborhoods with high chlamydia rates. Jail screening started immediately before chlamydia declines among young females at a clinic serving neighborhoods with high jail testing density. These programs might help reduce community prevalence and racial/ethnic disparities in STDs. PMID- 18418299 TI - Estimates of Chlamydia trachomatis infections among men: United States. AB - OBJECTIVE: To describe the epidemiology of genital Chlamydia trachomatis infections among men in the United States. STUDY DESIGN: Data from the notifiable disease case surveillance system, the National Health and Nutrition Examination Survey (NHANES), the National Longitudinal Study of Adolescent Health (AddHealth), the National Job Training Program, the Men Having Sex with Men (MSM) Prevalence Monitoring Project, and adult and juvenile corrections facilities were used to summarize national chlamydia case and prevalence rates. Data were stratified by age and race/ethnicity. RESULTS: In 2005, 232,781 chlamydia cases among men were reported, corresponding to a rate of 161.1 cases per 100,000 men, an increase of 43.5% compared with the case rate in 2001 (112.3). Population based chlamydia prevalence rates from NHANES (1999-2002) were highest among men aged 20 to 29 years (3.2%); men aged 18 to 26 years participating in AddHealth (2001-2002) had a 3.7% prevalence rate. Rates were highest among black men in both NHANES (5.3%) and AddHealth (11.1%). The prevalence rate among men (aged 16 24 years) participating in the National Job Training Program was 8.1%. Among MSM, the 2005 median urethral chlamydia prevalence rate was 6%. Overall, chlamydia rates were highest in adult corrections facilities; the 2005 positivity rate among men aged 21 to 25 years was 7.8%. In juvenile corrections facilities, the 2005 positivity rate among men aged 15 to 17 years was 6.7%. CONCLUSIONS: Rates of genital C. trachomatis infections among men are persistently high, particularly among men entering the National Job Training Program and men in corrections facilities. The burden of disease is generally highest among young men and black men. PMID- 18418300 TI - Male circumcision and women's risk of incident chlamydial, gonococcal, and trichomonal infections. AB - BACKGROUND: Male circumcision (MC) decreases the risk of human immunodeficiency virus (HIV) acquisition in men. We explored associations between MC of the primary sex partner and women's risk of acquisition of chlamydial (Ct), gonococcal (GC), or trichomonal (Tv) infections. METHODS: We analyzed data from a prospective study on hormonal contraception and incident human immunodeficiency virus/sexually transmitted infection (STI) among women from Uganda, Zimbabwe, and Thailand. At enrollment and each follow-up visit, we collected endocervical swabs for polymerase chain reaction identification of Ct and GC; Tv was diagnosed by wet mount. Using Cox proportional hazards models, we compared time to STI acquisition for women according to their partner's MC status. RESULTS: Among 5925 women (2180 from Uganda, 2228 from Zimbabwe, and 1517 from Thailand), 18.6% reported a circumcised primary partner at baseline, 70.8% reported an uncircumcised partner, and 9.7% did not know their partner's circumcision status. During follow-up, 408, 305, and 362 participants had a first incident Ct, GC, or Tv infection, respectively. In multivariate analysis, after controlling for contraceptive method, age, age at coital debut, and country, the adjusted hazard ratio (HR) comparing women with circumcised partners with those with uncircumcised partners for Ct was 1.25 [95% confidence interval (CI) 0.96-1.63]; for GC, adjusted HR 0.99 (95% CI 0.74-1.31); for Tv, adjusted HR 1.05 (95% CI 0.80-1.36), and for the 3 STIs combined, adjusted HR 1.02 (95% CI 0.85-1.21). CONCLUSIONS: MC was not associated with women's risk of acquisition of Ct, GC, or Tv infection in this cohort. PMID- 18418301 TI - Association of Mobiluncus curtisii with recurrence of bacterial vaginosis. PMID- 18418303 TI - Risk factors for gonorrhea among heterosexuals--San Francisco, 2006. AB - GOAL: In San Francisco, coinciding with increases in the western United States, we observed substantial gonorrhea increases among young heterosexuals during 2003 2005. We conducted a case-control study to identify intervention strategies for prevention and control. STUDY DESIGN: We interviewed case patients with gonorrhea during February-July, 2006 and control subjects at the local Department of Motor Vehicles. We included sexually active heterosexuals aged 15 to 35 years in sex stratified analyses. RESULTS: We interviewed 225 persons: 24 male and 28 female case patients and 98 male and 75 female control subjects. In multivariable analysis adjusting for black race and multiple partners among men, black race [adjusted odds ratio (AOR), 5.1; 95% confidence interval (CI), 1.7-15.0], having had multiple partners (AOR, 3.1; 95% CI, 1.1-8.5), having had an anonymous partner (AOR, 6.4; 95% CI, 1.9-21.4), and a long-term partnership (AOR, 0.3; 95% CI, 0.1-0.9) were associated with gonococcal infection. Among women, after adjustment for age, multiple partners, and black race (subject or partner), being black or having a black partner (AOR, 6.9; 95% CI, 2.2-21.8), having had a recently incarcerated partner (AOR, 6.2; 95% CI, 1.0-38.4), or meeting partners on the street (AOR, 19.0; 95% CI, 2.0-179.0) were associated with gonococcal infection. CONCLUSIONS: Demographic and behavioral factors increase risk for gonorrhea among heterosexuals in San Francisco with partner characteristics being particularly important. Prevention and control efforts are focusing on blacks and incarcerated populations using street-based outreach and expanded screening and treatment. PMID- 18418302 TI - Antimicrobial susceptibility profile of resistance phenotypes of Neisseria gonorrheae in India. AB - BACKGROUND AND OBJECTIVES: Regular monitoring of antimicrobial resistance of Neisseria gonorrheae needs to be established in the country. The aim of the study was to determine the trend of resistance phenotypes of N. gonorrheae in India, and to assess the susceptibility profile of resistance phenotypes. METHODS: Antimicrobial susceptibilities of N. gonorrheae isolates to penicillin, tetracycline, ceftriaxone, ciprofloxacin, and spectinomycin were determined by in terms of minimum inhibitory concentrations using Etest. World Health Organization reference strains were used as control strains. Phenotypic characterization of N. gonorrheae isolates was carried out on the basis of plasmid and chromosomally mediated resistance to penicillin and tetracycline. RESULTS: Of 382 isolates, 172 (45.0%) belonged to 6 resistance phenotype i.e., penicillinase-producing N. gonorrheae (PPNG) (16.5%), tetracycline resistant N. gonorrheae (TRNG) (8.9%), PPNG-TRNG (4.7%), chromosomally mediated tetracycline resistant N. gonorrheae (11.2%), and chromosomally mediated penicillin resistant N. gonorrheae (3.7%). Chromosomally mediated penicillin and tetracycline resistant N. gonorrheae isolates were not detected. Chromosomally mediated penicillin resistant N. gonorrheae and chromosomally mediated tetracycline resistant N. gonorrheae decreased from 2002 to 2006 whereas TRNG strains increased during the study period. Ciprofloxacin resistant and ceftriaxone less sensitive strains were more frequent among the resistance phenotypes. CONCLUSIONS: The high rates of plasmid and chromosomally mediated resistance to penicillin, tetracycline along with high rates of resistance to ciprofloxacin among all the resistance phenotypes underlines the necessity for continuous surveillance of antimicrobial resistance to help in controlling the spread of gonorrhoea. PMID- 18418304 TI - Implications of intensive glycemic control on cardiovascular disease: early reports from the ACCORD and ADVANCE Trials. PMID- 18418305 TI - An evidence-based approach to femoral arterial access and closure. AB - Percutaneous arterial access is performed in up to 10 million patients worldwide annually. The predominant techniques have been largely unchanged for a half century, but an evolving evidence base over the past several years suggests that major improvements in access and a reduction in complication rates may be accomplished with a better appreciation of anatomy and the use of fluoroscopy or ultrasound guidance. Vascular closure is still predominantly by manual compression; meta-analyses comparing vascular closure techniques have not clearly demonstrated a difference between manual compression and vascular closure device complication rates. Although vascular closure devices are associated with earlier hemostasis and time to ambulation, their cost and potential associated complications limit use to roughly 30% of cases in the United States and single digit percentages in the rest of the world. PMID- 18418306 TI - Histopathologic observations of femoral closure devices: understanding the differences. AB - Arteriotomy closure devices have been widely employed to expedite ambulation in patients undergoing diagnostic coronary angiography or percutaneous angioplasty. As compared with manual compression, these devices decrease time to complete vessel closure, allow earlier ambulation, and improve overall patient comfort. When complications occur, they are often severe and include vascular occlusion, significant bleeding, or inflammatory changes. Animal models have been used to evaluate the impact of these devices. In animal studies, the initial procedure seems to incite mild to moderate vascular spasm that resolves at an undetermined time after the initial procedure. Inflammation occurs early and is most prominent in suture-based and collagen-based procedures compared with extravascular clip procedures. At late time points (after 30 to 60 days), however, no significant differences in angiographic appearance of the vessels could be determined. PMID- 18418307 TI - Early detection and integrated management of dysglycemia in cardiovascular disease: a key factor for decreasing the likelihood of future events. AB - Dysglycemia is widespread among patients with coronary artery disease. It is indeed more common than normoglycemia in these patients. Coexistence of cardiovascular disease and dysglycemia presents significant health risks, and evidence suggests that both conditions should be treated early to reduce the development of complications. Guidelines recommend testing for prediabetes and type 2 diabetes in the cardiology setting and highlight the use of therapies that treat metabolic and cardiovascular risk factors. Blood glucose levels have previously been the interest of diabetologists, but modern integrated management approaches should include assessment by a cardiologist. We propose that postprandial blood glucose testing be carried out routinely in all patients with coronary artery disease, and that newly diagnosed dysglycemia be actively managed. PMID- 18418309 TI - Safety and efficacy of drug-eluting stents: on-label and off-label perspectives. AB - Favorable outcomes in multiple randomized trials have resulted in the widespread adoption of drug-eluting stents (DES) during percutaneous coronary intervention (PCI). However, reports of increased stent thrombosis-possibly with increased rates of late death and myocardial infarction (MI)-along with the requirement of an extended course of clopidogrel with DES, have resulted in uncertainties as to which patients should receive DES instead of bare-metal stents (BMS). In most patient and lesion subsets, DES significantly reduce neointimal proliferation (resulting in decreased angiographic restenosis), recurrent angina and ischemia, and the need for subsequent revascularization procedures. DES "off-label" indications include use in patients with multiple lesions and multiple vessels, lesions showing long diffuse disease, very small or very large vessels, true bifurcation lesions, thrombotic lesions, and conditions such as acute MI and chronic total occlusions. For now, pending more data, the risks and benefits of DES for off-label indications must be carefully considered on an individual patient basis. PMID- 18418308 TI - Targeting the kidney in acute decompensated heart failure: conventional diuretics and renal-acting vasodilators. AB - A mainstay of therapy for congestive heart failure has been the use of potent diuretic agents, such as furosemide, that target the kidney to enhance sodium and water excretion. Although furosemide is widely used to treat the symptoms of acute decompensated heart failure (ADHF), the consequent activation of the renin angiotensin-aldosterone system may limit the natriuretic response by reducing the glomerular filtration rate. In addition, excessive diuresis may reduce cardiac preload and result in systemic hypotension, which reduces renal perfusion pressure and prerenal azotemia and raises levels of blood urea nitrogen. In order to preserve and/or enhance renal function in ADHF, especially with agents such as conventional diuretics and vasodilators, an understanding of intrarenal factors that may protect the kidney may provide a direction for optimal use of current therapies and also lead to newer therapeutic strategies. Vasodilators, especially those that are linked to cGMP activation, may provide an alternative approach. PMID- 18418311 TI - Treatment of intractable angina pectoris utilizing spinal cord stimulation. AB - Intractable angina pectoris affects approximately 5% to 15% of patients with ischemic heart disease. Current treatment options for refractory angina can be divided into 3 groups: pharmacological, nonpharmacological noninvasive, and invasive. The newest pharmacological treatment option for intractable angina pectoris is ranolazine. Non-pharmacological, noninvasive treatment options include enhanced external counterpulsation and transcutaneous electrical nerve stimulation. Invasive treatment options include revascularization procedures: coronary artery bypass grafting, percutaneous transluminal coronary angioplasty, and percutaneous laser revascularization. Spinal cord stimulation (SCS) as a palliative intervention for refractory angina has been underutilized in the United States. This case review describes application of SCS in a 43-year-old woman with a 10-year history of symptomatic ischemic heart disease who was unresponsive to all available treatment options for intractable severe chest pain. Following spinal cord stimulator placement, the patient reported no further angina, discontinued nitroglycerine, had improved sleep quality, and resumed full time employment. PMID- 18418312 TI - Optimizing outcomes in CT imaging. PMID- 18418313 TI - Iodinated contrast media and the kidney. AB - One of the principal complications of radiographic procedures utilizing intravascular iodinated contrast media is acute kidney injury. Although several clinical and procedural factors impact a patient's risk for contrast-induced acute kidney injury (CIAKI), substantial attention has been focused on the relationship between the type of contrast agent used and renal injury. Multiple contrast agents are available for clinical use, each defined by a series of physicochemical properties. The evolution from high osmolal to low osmolal and, more recently, iso-osmolal contrast media has led to several clinical trials and meta-analyses comparing the nephrotoxicity of different contrast agents. This article summarizes the physicochemical properties that define and differentiate iodinated contrast media, discusses the purported relationship between these properties and kidney injury, and describes the salient findings of clinical trials and meta-analyses that have compared the nephrotoxic effects of contrast agents. Although ongoing and future studies will further elucidate our understanding of the relationship between iodinated contrast and risk for CIAKI, a sound understanding of the currently available data will help inform evidence based decisions on the use of these agents in clinical practice. PMID- 18418314 TI - Contrast media: procedural capacities and potential risks. AB - Contrast media are known to have transient hemodynamic properties that can influence a patient's clinical status, including heart rate variability and blood pressure. These changes have the potential to impact the diagnostic quality of CT scans. Although most patients are able to receive contrast media without significant adverse reactions, events occur in a minority of cases. These reactions range from mild discomfort (injection-associated pain and heat sensation) to more significant cardiac, renal, and hypersensitivity reactions. The incidence of adverse reactions varies with the type of contrast media used, and several randomized trials have elucidated the cardiac and renal differences among agents. Risk factors for contrast-induced acute kidney injury (CIAKI) have been established, with baseline kidney disease amplified by the presence of diabetes constituting the highest-risk patient group. Strategies for preventing CIAKI include antioxidant therapy, hydration regimens, and choice of contrast agents. Enhanced knowledge on the part of physicians and medical personnel regarding the properties and potential side effects of iodinated contrast agents should lead to improved patient safety and efficacy when performing radiologic examinations. PMID- 18418315 TI - Priceless clinical pearls in the performance of cardiac CT. AB - Cardiovascular CT represents an important innovation in cardiac imaging as a noninvasive modality for the diagnosis of coronary artery disease. As a screening examination, coronary CT has the ability to identify the presence, extent, location, and severity of coronary artery disease in patients at risk. As a diagnostic examination in patients with chest discomfort and dyspnea, it can not only evaluate for the presence of coronary artery disease but also evaluate the pulmonary vasculature and aorta. The ability to perform high-quality cardiac CT requires a combination of technical expertise and knowledge of cardiac anatomy. From our experience having performed over 6000 cardiac CTs, we provide the reader with a number of clinical and technical pearls that will enhance his or her ability to perform high-quality studies even in the more challenging patient. PMID- 18418316 TI - Interventional radiology procedures: addressing the needs of the cardiovascular patient. AB - Interventional radiologists perform a wide range of procedures on virtually every organ system in the body. Technological advances continue to expand the repertoire of procedures in this field, challenging the frontiers of medical science and individual expertise. With cardiovascular disease being the number 1 cause of death in the United States and an aging population before us, physicians who perform minimally invasive procedures must be knowledgeable regarding unique aspects of cardiovascular management as it pertains to the catheterization laboratory. Specific areas addressed in this review include anesthesia, hypertension, and chronic kidney disease. PMID- 18418317 TI - Acute pyogenic diskitis (spondylodiskitis) in adults. AB - Most cases of back pain are the result of degenerative changes in the spine or are related to musculoskeletal elements. Pyogenic infections of the back can be subcategorized into cases involving the paraspinal epidural space, vertebral bodies, or the intervertebral disk spaces. Any region of the spine may be the site of diskitis, although the process most commonly involves the lumbar spine. Most cases of diskitis are managed with conservative therapy, including antibiotics and spinal immobilization using braces or corsets. Surgical therapy is generally reserved for patients with neurological complications, spinal instability, or progressive spinal deformity or those who fail to respond clinically to antibiotic therapy alone. PMID- 18418318 TI - Peripheral neuropathies associated with monoclonal gammopathies of undetermined significance. AB - Monoclonal gammopathies (MGs) or plasma cell dyscrasias (PCDs) are hematologic disorders that may affect peripheral nerves. An MG is a proliferation of a single clone of neoplastic or nonneoplastic plasma that usually secretes a monoclonal protein (M-protein) serum or urine. If a diagnosis of monoclonal gammopathy of undetermined significance (MGUS) is established, a sudden increase in M-protein levels can indicate malignant transformation of a benign PCD. Roughly 50% of MGUS neuropathies are associated with an IgM gammopathy and the remaining 50% with IgG and IgA-MGUS-associated neuropathies. MGUS is the most common of the PCDs associated with neurologic disorders, which are easily approached clinically by classifying them as IgM or non-IgM types. PMID- 18418319 TI - Neuromuscular diseases. PMID- 18418320 TI - An unusual case of multiple discrete brain lesions. AB - The case of a patient with multiple enhancing lesions in the brain is presented, along with discussion of the differential diagnosis of granulomatous lesions of the brain, and a brief review of the essential features and neurological complications of the eventual diagnosis. Aspects of the disease of interest to neurologists are emphasized. PMID- 18418322 TI - Sleep disorders. PMID- 18418321 TI - Language impairment: stepwise or slowly progressive? AB - MRI showed ischemic lesions of the left hemisphere in a patient with a worsening language disturbance that by history could be caused either by ischemia or by a focal degenerative brain disorder. A positron emission tomography study indicated the presence of a degenerative disorder, the most common cause of which is tau positive pathology. PMID- 18418323 TI - Epilepsy. PMID- 18418324 TI - Clinical trials in epilepsy: the SANAD study. PMID- 18418327 TI - The interactions between sleep and epilepsy. AB - The interactions between sleep and epilepsy have been recognized for centuries. The electroencephalogram and more recently the polysomnogram have helped us understand these interactions. Certain epilepsy syndromes are more common during sleep, and frequently a sleep disorder is misdiagnosed as epilepsy. Management of epilepsy includes improvements in sleep quality and quantity, and aggressive treatment of epilepsy has a positive impact on sleep disorders. PMID- 18418328 TI - The Euro-American Rehabilitation Focus: a cultural bridge across the ocean. PMID- 18418329 TI - Practice of physical medicine and rehabilitation on both sides of the Atlantic: differences and the factors that drive them. PMID- 18418330 TI - Physical and rehabilitation medicine: comments related to the White Book on Physical and Rehabilitation Medicine in Europe. PMID- 18418331 TI - Effects of walking with a cane on balance and social participation among community-dwelling post-stroke individuals. AB - AIM: Walking devices such as canes and frames are often prescribed to post-stroke individuals to enhance their balance and walking, and thus facilitate community reintegration and social participation. The aim of this study was to observe social participation and balance in post-stroke hemiparetic patients and compare the performance of those walking with a cane (AD group) and without a cane (WAD group). The relationships between cane usage and activity participation and balance were also studied. METHODS: In this ex post facto research study, balance and social participation were evaluated using the Berg Balance Scale (BBS) and the Craig Handicap Assessment and Reporting Technique (CHART) in 50 individuals with hemiparesis secondary to first incidence hemispheric stroke. The study included 25 AD subjects (mean age 59.88+/-12.04 years) and 25 WAD subjects (mean age 55.84+/-11.30 years) who were consecutively recruited from the physiotherapy units of Korle Bu Teaching Hospital (KBTH) and the 37 Military Hospital in Accra (Ghana). Before recruitment into the study, both groups of subjects were matched for age, height, weight, BBS and CHART scores at the point of discharge from the inpatient units of the two hospitals, the duration of stroke, and the duration of physiotherapy post-stroke. RESULTS: Mann-Whitney U statistics showed that the AD group had comparatively higher mean participation scores (U=22.37; P=0.00) and lower mean balance performance (BBS score U=30.68; P=0.00) than their WAD counterparts. Spearman's correlation coefficient also revealed a significant negative correlation between the BBS and CHART scores in both groups (AD rho= 0.063; P=0.00 and WAD rho=-0.037; P=0.05). CONCLUSION: These findings indicate that post-stroke individuals who used a cane to aid in walking had poorer balance and less social participation than their age-matched counterparts who walked unaided. As balance improved (higher BBS score), participation also improved (lower CHART score) in both groups. In spite of the small sample size, the outcome of this study suggests that prescription of a cane for a post-stroke individual should be carefully considered by the physiotherapist, especially if the focus of rehabilitation includes restoration of balance functions and social participation. PMID- 18418332 TI - Spatio-temporal and kinematic asymmetry ratio in subgroups of patients with stroke. AB - AIM: To determine the spatio-temporal and kinematic gait asymmetry in hemiparetic patients after stroke, and to compare the subgroups in terms of asymmetry ratio. METHODS: Spatio-temporal and kinematic characteristics of gait in 100 patients (37 females, 63 males) with hemiparesis after stroke were retrospectively evaluated. The mean+/-SD age was 57.9+/-12.1 years (range 18-80) and time since stroke was 5.75+/-5.9 months (range 1-36). Subgroups were based on age, gender, side of paresis, lesion type, motor recovery level, sensory status, time since stroke and walking velocity. RESULTS: The older patients (65 years) had a higher temporal asymmetry in terms of single-support time whereas the younger patients (<65 years) had a higher kinematic asymmetry in terms of ankle joint kinematics at both stance and swing phases (P<0.05). In the poor motor recovery group, asymmetry ratios of step length, hip and knee extension at stance, and ankle kinematics were significantly higher than those of the good motor recovery group (P<0.05). In the slow walking group, step length showed a greater asymmetry than in the fast walking group (P<0.05). Gait asymmetry was similar among the patients when they were grouped according to their gender, side of paresis, lesion type, proprioception, and time since stroke. CONCLUSION: For better interpretation of quantitative gait data and to offer appropriate rehabilitation programs, clinicians should consider that spatio-temporal and kinematic asymmetry might vary according to age, motor recovery level and walking speed of hemiparetic patients after stroke. PMID- 18418333 TI - The diagnostic importance of exercise testing in developing appropriate rehabilitation programmes for patients following transfemoral amputation. AB - AIM: The aim of this study was to determine whether an arm ergometry exercise test can successfully predict a clinician's decision for prosthetic fitting, and to define what other factors are also important predictors of a patient's ability to walk with a prosthesis. METHODS: The study included 63 patients who had undergone transfemoral amputation due to peripheral vascular disease and who had been admitted for primary inpatient rehabilitation. Participating patients went through standard admission procedures and performed an intermittent sub maximal arm ergometry exercise test in the first week after admission consisting of a two minute exercise routine with a one-minute pause in-between by means of the Angio V2. The work loads started at 10 W and were increased by 10 W at each level. The cranking rate was 50 to 55 rpm. RESULTS: The patients reached an output level of 20 W on average. Those patients selected for fitting with a prosthesis achieved higher levels on the exercise stress test (26 versus 12 W, P<0.000). The results of the exercise stress test, the 6-minute walking test and the functional independence measures (FIM) at admission, as well as their age and gender, allowed for successful discrimination between patients who were fitted with a prosthesis and those who were not (P<0.000). The best discriminator was that of the results achieved on the exercise stress test. Using the five previously mentioned variables, 83.6% of the patients were successfully classified as to whether or not they were fitted for a prosthesis on the basis of our clinical decision. CONCLUSION: The exercise stress test is a good predictor of prosthetic fitting. It also anticipates the possibility of cardiovascular complications during rehabilitation programmes. Combined with the 6-minute walk, the FIM, and age, it may be an important criteria to consider in making the decision about the appropriateness of prosthetic fitting. PMID- 18418334 TI - Reduced aerobic capacity in patients with severe osteoporosis: a cross sectional study. AB - AIM: It has been previously shown that pulmonary function (PF) is significantly diminished in patients with osteoporosis (OP). But there are few data about the relationship between PF and aerobic capacity of osteoporotic patients and the severity of thoracic kyphosis and time since the diagnosis of OP. The aim of the present study was to investigate the resting spirometric values and cardiopulmonary test (CPET) results of women with osteoporosis and to evaluate the effects of the various degrees of OP on these parameters. METHODS: Fifty six outpatient subjects were included in the study. All patients underwent a standardized interview, physical examination, bone mineral density (BMD), anteroposterior and lateral x-rays of thoracic spine, resting PF test and CPET evaluation. To evaluate the effects of the severity of osteoporosis on these parameters patient group divided according to diagnosis time of OP, degree of kyphosis, and spinal deformity index. Demographic and clinical data were compared between the groups with the use of independent-sample t test analysis (two groups) and analysis of variance (ANOVA) was used to estimate the between-group differences and changes by severity of osteoporosis and regression analyses to find predictors for changes. Correlation coefficients were used to examine the relationship between variables. RESULTS: According to diagnosis time, newly diagnosed groups; according to degree of kyphosis, the kyphotic groups; according to spinal deformity index, the higher spinal deformity index groups showed statistically significant and declining results in PF tests and CPET parameters. CONCLUSION: This study pointed out a significant impaired PF, aerobic capacity and a serious deconditioning for various reasons in these OP patients Therefore, the evaluation of CPET should be included in the management of OP patients and in these patients ventilatory muscle training and aerobic exercises may offer a potential therapeutic adjunct to current OP therapies in the future. PMID- 18418335 TI - Nutritional counselling in disabled people: effects on dietary patterns, body composition and cardiovascular risk factors. AB - AIM: Disabled persons are frequently affected by nutritional status impairment, consequent to quantitative and qualitative inadequacy of diet and physical inactivity, resulting in a significant reduction of fat-free mass and bone mineral density (BMD), and an over-expression of fat mass and an increased number of biochemical risk factors for chronic degenerative diseases. The aim of this study was to analyse the applicability and the efficacy of a nutritional counselling intervention in order to improve dietary intake and nutritional status in disabled people. METHODS: Thirty-seven disabled subjects (24 with physical disability and 13 with both mental retardation and physical disability; age 33.5+/-9.2 years) underwent an assessment of nutritional status, and an intervention with nutritional counselling was proposed to each patient for one year. Anthropometric measurements, indirect calorimetry, dual-energy X-ray absorptiometry, dietary intake, and biochemical analysis at baseline (T0) and after one year (T1) of counselling intervention were performed. RESULTS: Sixty five percent of patients dropped out. Overall, no significant improvement in cardiovascular risk factors, body composition and dietary patterns was reported at T1 in completer subjects. Six subjects who were obese or overweight at T0, reported significant weight and fat mass (FM) reduction at T1 (P=0.01 and P=0.00, respectively). CONCLUSION: Nutritional counselling seems to be ineffective and poorly applicable to disabled people. Further studies should be directed towards a treatment program associated with careful screening, motivation analysis, and follow-up in this patient population. PMID- 18418336 TI - Cycling induced by functional electrical stimulation improves the muscular strength and the motor control of individuals with post-acute stroke. Europa Medicophysica-SIMFER 2007 Award Winner. AB - AIM: The aim of this study was to investigate the effectiveness of cycling induced by functional electrical stimulation (FES) in patients with postacute stroke. METHODS: Twenty postacute inpatients were recruited and were randomly shared in a control group (56+/-9.2 years old, 50.8+/-24.5 days post-stroke) performing the standard rehabilitation (SR) and a FES group (51+/-12 years old, 56.1+/-22.8 days post-stroke) performing FES cycling in addition to SR. Both the groups performed 3 hours of rehabilitation per day for 4 weeks. The FES cycling was applied daily for 35 minutes and quadriceps, hamstring, gluteus maximus and tibialis anterior of both the legs were stimulated. The two groups were compared by the following outcome measurements before and after treatment: maximum isometric voluntary contraction (MVC) of quadriceps, walking and sit-to-stand ability, motricity index, upright motor control test and trunk control test. RESULTS: After the treatment, the U-Mann-Whitney test demonstrated that the FES group produced a significantly higher increase of the muscular force produced by both the quadriceps during MVC with respect to the control group (P<0.05). Seventy percent of FES patients learned how to perform the sit to stand movement with three different rising speeds while no control patients develop the ability to perform the task properly. CONCLUSION: Rehabilitation including FES cycling was more effective in promoting muscle strength and motor recovery of the lower extremity than therapist-assisted SR alone. Tests on an enlarged number of patients are necessary for generalization before proposing FES cycling in the clinical rehabilitation of post-acute stroke patients. PMID- 18418337 TI - Rehabilitation of adolescent idiopathic scoliosis: results of exercises and bracing from a series of clinical studies. Europa Medicophysica-SIMFER 2007 Award Winner. AB - AIM: Rehabilitation of adolescent idiopathic scoliosis (AIS) requires a careful choice from among the possible treatments, such as bracing and exercises, according to the patient's needs. According to the literature, there is little evidence regarding the efficacy of these rehabilitation instruments. During the past few years, a full series of studies has been carried out to investigate their efficacy. The aim of this paper was to summarize all these results. METHODS: Three systematic reviews (two on exercises and one on manual therapy), and four cohort prospective studies were performed. The prospective studies included two trials with a prospective control group on exercises (one to avoid bracing and one in preparation to bracing) and two trials with retrospective control group on a new brace developed by the Authors (Sforzesco brace and SPoRT concept of correction versus Lyon brace and Risser cast). RESULTS: Results show that in literature there is proof of level 1b on exercises but no studies on manual therapy. High quality exercises like Scientific Exercises Approach to Scoliosis (SEAS) have more efficacy than usual physiotherapy, significantly reducing brace prescription in one year from 25% of cases to 6%. Moreover, such exercises help to obtain the best results in bracing first correction. The Sforzesco brace has proved to have more efficacy than the Lyon brace, whereas it has the same efficacy--but reduced side effects and impact on quality of life- than the Risser brace. CONCLUSION: With an efficient management of data collection, it is possible to develop a set of studies aimed at verifying the efficacy of clinical daily rehabilitation approaches. PMID- 18418338 TI - The treatment of adolescent idiopathic scoliosis (AIS) according to present evidence. A systematic review. AB - Traditionally, the treatment options for adolescent idiopathic scoliosis (AIS), the most common form of scoliosis, are exercises; in-patient rehabilitation; braces and surgery. The outcomes of treatments are usually compared with the natural history or observation (non-intervention). The aim of this paper was to provide a synopsis of all treatment options in the light of evidence based practice (EBP). A systematic review was carried out using the most encompassing databases available. Literature has been searched for the outcome parameter ''rate of progression'' and only prospective controlled studies that have considered the treatment versus the natural history have been included. The search strategy included the following terms: ''adolescent idiopathic scoliosis''; ''idiopathic scoliosis''; ''natural history''; ''observation''; ''physiotherapy''; ''physical therapy''; ''rehabilitation''; ''bracing''; ''orthotics'' and ''surgery''. Prospective short-term studies have been found to support outpatient physiotherapy. One prospective controlled study was found to support scoliosis in-patient rehabilitation (SIR). One prospective multi-centre study, a long-term prospective controlled study and a meta-analysis have been found to support bracing. No controlled study, neither short, mid nor long-term, was found to reveal any substantial evidence to support surgery as a treatment for this condition. There is some evidence supporting the conservative treatment for AIS. No substantial evidence has been found in terms of prospective controlled studies to support surgical intervention. In light of the unknown long term effects of surgery, a randomised controlled trial (RCT) seems necessary. Due to the presence of evidence to support conservative treatments, a plan to compose a RCT for conservative treatment options seems unethical. But it is also important to conclude that the evidence for conservative treatments is weak in number and length. PMID- 18418339 TI - The term diplegia should be enhanced. Part I: a new rehabilitation oriented classification of cerebral palsy. AB - The classification systems for cerebral palsy (CP) need to be continuously updated, according to specific aims and to significant changes observed over the years in the panorama of CP. A simplification of CP categories, abandoning the use of the term diplegia, has been recently suggested. Conversely, in this paper a new proposal for classification of CP is briefly presented, where special attention is given to diplegia which is suggested to be divided into four main clinical forms, according to the patterns of walking observable in these subjects. The proposed classification was applied to a large population of 213 subjects with diplegia, among 467 cases of CP admitted to two reference centres for this disorder. The relative incidence of the four forms is reported. The adopted classification criteria, based on a primary ability of professionals working in rehabilitation, i.e. observation capacity, makes this approach simple and easy to use at all levels of the rehabilitation services for CP. PMID- 18418340 TI - The term diplegia should be enhanced. Part II: contribution to validation of the new rehabilitation oriented classification. AB - Recent proposals of classification for cerebral palsy (CP), mainly revised for epidemiological purposes, suggest to abandon the use of the term diplegia. Conversely, in this paper data are presented to support the proposal to maintain the distinction between spastic tetraplegia and diplegia, and to subdivide this latter according to four main clinical patterns of walking observable in these children. This proposal of classification was validated by testing a group of 467 subjects with CP, of whom 213 with diplegia and 115 with tetraplegia, consecutively admitted between January 2005 and December 2006 to two national reference centers for this disability. The results were compared with findings obtained by other methods of classifying gross and fine motor function and associated disorders. The subjects with tetraplegia strongly differ from those of diplegia, both for motor functions and for other disabilities. The four main walking patterns of spastic diplegia were easily recognizable and observers were able to assign most of the subjects to one form of the classification. Significant correlations between walking forms of diplegia and distribution of Gross Motor Function Classification System (GMFCS) levels were found. Some of the forms significantly differ also for fine motor and mental disability. These findings suggest that in clinical practice the category of diplegia not only can be kept as a separate form of CP, but it may be enhanced, through the identification of different subcategories of children, divided according to their walking patterns. PMID- 18418341 TI - The term diplegia should be enhanced. Part III: inter-observer reliability of the new rehabilitation oriented classification. AB - AIM: The aim of this study was to validate a recent classification of gait in children with the spastic diplegic form of cerebral palsy (CP) by checking the reliability of different scorers in assigning subject walking performance to one of the four specific patterns described in the classification. METHODS: The gait patterns of 50 children and adolescents with CP (23 males, 27 females; age range 3-17 years) were selected among patients whose videos were stored in the archives of the Pisa and Reggio Emilia Hospitals. Only video recordings of gait with homogeneous features (duration of at least 90 s, simultaneous recordings on sagittal and frontal views, and other criteria) were taken for examination. The videos were blindly scored using an observational gait scale, at first by two of the authors of the classification system (defined as ''maximum experts''), then by ten expert observers, and finally by 206 professionals of rehabilitation after a one-day training on the classification. Cohen's kappa statistics (k) and intra class correlations (ICC) were calculated. RESULTS: Kappa and ICC indicate an almost perfect agreement both between the two maximum experts and among the ten expert observers. Good results were also obtained in the group of one-day trained scorers. Only a few cases were assigned to the ''unclassified'' category. The profession of the observer (doctor or therapist) and previous knowledge of the classification had no significant influence on reliability scores. CONCLUSION: The results suggest that the proposed classification can be reliably applied, even utilizing short video recordings, to arrange diplegic children into different patterns. Further studies are needed to validate the use of this classification system for clinical and research aims. PMID- 18418342 TI - To Google or not to Google, this is the question. AB - To Google or not to Google has become in the latest years a big question for researchers: immediate answers, but often thousands; sometimes pertinent, sometimes not reliable or too commercial-oriented. No doubt that Google always ''finds'' something and sometimes it is the only way, or it retrieves real pearls but where does it search? How can a researcher refine or limit the search? That is why we'll explore some Google features not so widely known, and other search engines with useful devices to perform a more efficient search in the biomedical field. PMID- 18418343 TI - Is planar imaging in radiology and nuclear medicine a viable option for the 21st century? PMID- 18418344 TI - Interim within-season estimate of the effectiveness of trivalent inactivated influenza vaccine--Marshfield, Wisconsin, 2007-08 influenza season. AB - During clinical trials, the efficacy of vaccination with inactivated influenza vaccines for the prevention of serologically confirmed influenza infection has been estimated as high as 70%-90% among healthier adults. However, the effectiveness of annual influenza vaccination typically is lower during those influenza seasons when a suboptimal match between the vaccine strains and circulating influenza strains is observed. For example, in a 4-year randomized study of influenza vaccine among healthy persons aged 1-65 years, the predominant strain was drifted from the vaccine strain in 2 of the 4 years. Inactivated vaccine effectiveness (VE) against culture-confirmed influenza ranged from 71% to 79% when the vaccine and circulating strains were suboptimally matched to 74% to 79% when the matches were well matched. In contrast, a 2-year study of inactivated influenza vaccine among healthy adults aged 18-64 years found no measurable VE during a year when a poorly matched strain circulated, but found VE of 86% against laboratory-confirmed influenza during the following year when the vaccine and circulating strains were well matched. Although laboratory data on the antigenic characteristics of circulating influenza viruses compared with vaccine strains are available during influenza seasons, estimates of VE usually have not been made until months after the conclusion of the season. This report summarizes interim results of a 2008 case-control study to estimate the effectiveness of trivalent inactivated influenza vaccine for prevention of medically attended, laboratory-confirmed influenza during the 2007-08 influenza season, when most circulating influenza A (H3N2) and B viruses were suboptimally matched to the vaccine strains. Despite the suboptimal match between two of three vaccine strains and circulating influenza strains, overall VE in the study population during January 21-February 8, 2008, was 44%. These findings demonstrate that, in any season, assessment of the clinical effectiveness of influenza vaccines cannot be determined solely by laboratory evaluation of the degree of antigenic match between vaccine and circulation strains. PMID- 18418345 TI - Rotavirus vaccination coverage and adherence to the Advisory Committee on Immunization Practices (ACIP)-recommended vaccination schedule--United States, February 2006-May 2007. AB - Worldwide, rotavirus is the leading cause of severe gastroenteritis in children aged <5 years. In February 2006, a new human-bovine rotavirus vaccine, RotaTeq (Merck & Co., Inc., Whitehouse Station, New Jersey), was recommended by the Advisory Committee on Immunization Practices (ACIP) for routine vaccination of U.S. infants. Three doses of RotaTeq are recommended at ages 2, 4, and 6 months. The first dose should be administered between ages 6 and 12 weeks, and vaccination should not be initiated for infants aged >12 weeks. Subsequent doses should be administered at 4-10 week intervals, with all doses administered by age 32 weeks. This schedule is consistent with the ages at which RotaTeq was administered during prelicensure trials, and ACIP has recommended that RotaTeq only be administered at the ages for which safety and efficacy data are available. In 1999, a previous rhesus-human rotavirus vaccine, RotaShield (Wyeth Laboratories, Inc., Marietta, Pennsylvania), was withdrawn voluntarily from the U.S. market by the manufacturer because it was associated with intussusception, a form of bowel obstruction. The greatest risk for intussusception was noted after the first dose of RotaShield. Data from a large-scale, prelicensure safety trial and postlicensure monitoring do not indicate an association between the current RotaTeq vaccine and intussusception. CDC assessed rotavirus vaccination coverage among U.S. infants during February 2006-May 2007 and examined adherence to the ACIP-recommended vaccination schedule. This report summarizes the results of that assessment, which indicated that, by May 15, 2007, nearly half of infants aged 3 months had received 1 dose of rotavirus vaccine, with the majority of doses administered according to ACIP recommendations. Health-care providers should remain vigilant in following the ACIP-recommended vaccination schedule for rotavirus vaccine. PMID- 18418346 TI - Laboratory-acquired vaccinia exposures and infections--United States, 2005-2007. AB - The last case of naturally acquired smallpox disease, caused by the orthopoxvirus variola virus (VARV), occurred in 1977, and the last laboratory-acquired case occurred in 1978. Smallpox was eradicated largely as the result of a worldwide vaccination campaign that used the related orthopoxvirus, vaccinia virus (VACV), as a live virus vaccine. Routine childhood vaccination for smallpox in the United States was terminated by 1972, but vaccination continues or has been reintroduced for specific groups, including laboratory workers who may be exposed to orthopoxviruses, members of the military, selected health-care workers, and first responders. Severe complications of VACV infection can occur, particularly in persons with underlying risk factors, and secondary transmission of VACV also can occur. VACV is used in numerous institutions for various research purposes, including fundamental studies of orthopoxviruses and use as a vector for the expression of foreign proteins (often antigens or immunomodulators) in eukaryotic cells and animal models. The widespread use of VACV for research has resulted in laboratory-acquired VACV infections, some requiring hospitalization. The current Advisory Committee on Immunization Practices (ACIP) guidelines recommend VACV vaccination for laboratory workers who handle cultures or animals contaminated or infected with nonhighly attenuated VACV strains or other orthopoxviruses that infect humans. This report describes five recent occurrences of laboratory acquired VACV infections and exposure and underscores the need for proper vaccination, laboratory safety, infection-control practices, and rapid medical evaluation of exposures in the context of orthopoxvirus research. PMID- 18418347 TI - Update: influenza activity--United States, September 30, 2007-April 5, 2008, and composition of the 2008-09 influenza vaccine. AB - This report summarizes U.S. influenza activity since September 30, 2007, the start of the 2007-08 influenza season, and updates the previous summary. Low levels of influenza activity were reported from October through early December. Activity increased from mid-December and peaked in mid-February. PMID- 18418348 TI - Compendium of animal rabies prevention and control, 2008: National Association of State Public Health Veterinarians, Inc. (NASPHV). AB - Rabies is a fatal viral zoonosis and a serious public health problem. The disease is an acute, progressive encephalitis caused by a lyssavirus. Although the United States has been declared free of canine rabies virus variant transmission, multiple viral variants are maintained in wild mammal populations, and there is always a risk of reintroduction of canine rabies. All mammals are believed to be susceptible to the disease, and for purposes of this document, use of the term "animal" refers to mammals. The recommendations in this compendium serve as a basis for animal rabies-prevention and -control programs throughout the United States and facilitate standardization of procedures among jurisdictions, thereby contributing to an effective national rabies-control program. This document is reviewed annually and revised as necessary. The most current version replaces all previous versions. These recommendations do not supersede state and local laws or requirements. Principles of rabies-prevention and -control are detailed in Part I; recommendations for parenteral vaccination procedures are presented in Part II; and all animal rabies vaccines licensed by the U. S. Department of Agriculture (USDA) and marketed in the United States are listed in Part III. PMID- 18418349 TI - Expression and function of the Delta-1/Notch-2/Hes-1 pathway during experimental acute kidney injury. AB - The Notch signaling pathway consists of several receptors and their ligands Delta and Jagged and is important for embryogenesis, cellular differentiation and proliferation. Activation of Notch receptors causes their cleavage yielding cytoplastic domains that translocate into the nucleus to induce target proteins such as the basic-loop-helix proteins Hes and Hey. Here we sought to clarify the significance of the Notch signaling pathway in acute kidney injury using a rat ischemia-reperfusion injury model and cultured NRK-52E cells. Analysis of the whole kidney after injury showed increased expression of Delta-1 and Hes-1 mRNA and protein along with processed Notch-2. Confocal microscopy, using specific antibodies, showed that Delta-1, cleaved Notch-2 and Hes-1 colocalized in the same segments of the injured renal proximal tubules. Recombinant Delta-1 significantly stimulated NRK-52E cell proliferation. Our study suggests that the Delta-1/Notch-2/Hes-1 signaling pathway may regulate the regeneration and proliferation of renal tubules during acute kidney injury. PMID- 18418350 TI - Transcription factor HNF1beta and novel partners affect nephrogenesis. AB - Heterozygous mutations of the tissue-specific transcription factor hepatocyte nuclear factor (HNF)1beta, cause maturity onset diabetes of the young (MODY5) and kidney anomalies including agenesis, hypoplasia, dysplasia and cysts. Because of these renal anomalies, HNF1beta is classified as a CAKUT (congenital anomalies of the kidney and urinary tract) gene. We searched for human fetal kidney proteins interacting with the N-terminal region of HNF1beta using a bacterial two-hybrid system and identified five novel proteins along with the known partner DCoH. The interactions were confirmed for four of these proteins by GST pull-down assays. Overexpression of two proteins, E4F1 and ZFP36L1, in Xenopus embryos interfered with pronephros formation. Further, in situ hybridization showed overlapping expression of HNF1beta, E4F1 and ZFP36L1 in the developing pronephros. HNF1beta is present largely in the nucleus where it colocalized with E4F1. However, ZFP36L1 was located predominantly in the cytoplasm. A nuclear function for ZFP36L1 was shown as it was able to reduce HNF1beta transactivation in a luciferase reporter system. Our studies show novel proteins may cooperate with HNF1beta in human metanephric development and propose that E4F1 and ZFP36L1 are CAKUT genes. We searched for mutations in the open reading frame of the ZFP36L1 gene in 58 patients with renal anomalies but found none. PMID- 18418351 TI - Progress in gene targeting: using mutant mice to study renal function and disease. AB - Genetic engineering in mice has provided much information about gene function in renal health and disease. This knowledge has largely come from conventional transgenic approaches. Recently, methods have been developed to control the cell type, timing and reversibility of target gene expression. Advances in identifying promoters conferring renal cell-specific gene regulation in vivo have greatly facilitated interpretation of gene targeting studies. Site-specific recombinases have permitted cell-specific knockout of genes; Cre is the preeminent recombinase, but recent progress with other recombinases, include Flp and PhiC31, will likely increase the usefulness of this class of enzymes. Temporally regulated gene expression, particularly using doxycycline- and tamoxifen inducible systems, holds great promise for avoiding developmental effects of gene mutations as well as facilitating comparison of the same animal's phenotype before and after gene modification. RNA interference is undergoing tremendous growth and has great potential for achieving gene knockdown quickly and reversibly. To date, however, the utility of these systems in modifying renal function in transgenic mice remains unproven. Finally, new gene targeting tools are in development that may substantially simplify generation of transgenic animals. This review discusses the state-of-the-art in gene targeting in the kidney, reviewing function, indications and limitations of the molecular biologic tools. PMID- 18418352 TI - Tubuloglomerular feedback: mechanistic insights from gene-manipulated mice. AB - Tubuloglomerular feedback (TGF) describes a causal and direct relationship between tubular NaCl concentration at the end of the ascending limb of the loop of Henle and afferent arteriolar tone. The use of genetically altered mice has led to an expansion of our understanding of the mechanisms underlying the functional coupling of epithelial, mesangial, and vascular cells in TGF. Studies in mice with deletions of the A or B isoform of NKCC2 (Na,K,2Cl cotransporter) and of ROMK indicate that NaCl uptake is required for response initiation. A role for transcellular salt transport is suggested by the inhibitory effect of ouabain in mutant mice with an ouabain-sensitive alpha1 Na,K-ATPase. No effect on TGF was observed in NHE2- and H/K-ATPase-deficient mice. TGF responses are abolished in A1 adenosine receptor-deficient mice, and studies in mice with null mutations in NTPDase1 or ecto-5'-nucleotidase indicate that adenosine involved in TGF is mainly derived from dephosphorylation of released ATP. Angiotensin II is a required cofactor for the elicitation of TGF responses, as AT1 receptor or angiotensin-converting enzyme deficiencies reduce TGF responses, mostly by reducing adenosine effectiveness. Overall, the evidence from these studies in genetically altered mice indicates that transcellular NaCl transport induces the generation of adenosine that, in conjunction with angiotensin II, elicits afferent arteriolar constriction. PMID- 18418353 TI - Approach to acute renal failure in biopsy proven myeloma cast nephropathy: is there still a role for plasmapheresis? PMID- 18418354 TI - Xenotransplantation: where are we in 2008? AB - Xenotransplantation holds promise to solve the ever increasing shortage of donor organs for allotransplantation. In the last 2 decades, major progress has been made in understanding the immunobiology of pig-into-(non)human primate transplantation and today we are on the threshold of the first clinical trials. Hyperacute rejection, which is mediated by pre-existing anti-alpha Gal xenoreactive antibodies, can in non-human primates be overcome by complement- and/or antibody-modifying interventions. A major step forward was the development of genetically engineered pigs, either transgenic for human complement regulatory proteins or deficient in the alpha1,3-galactosyltranferase enzyme. However, several other immunologic and nonimmunologic hurdles remain. Acute vascular xenograft rejection is mediated by humoral and cellular mechanisms. Elicited xenoreactive antibodies play a key role. In addition to providing B cell help, xenoreactive T cells may directly contribute to xenograft rejection. Long-term survival of porcine kidney- and heart xenografts in non-human primates has been obtained but required severe T and B cell immunosuppression. Induction of xenotolerance, e.g. through mixed hematopoietic chimerism, may represent the preferred approach, but although proof of principle has been delivered in rodents, induction of pig-to-non-human primate chimerism remains problematic. Finally, it is now clear that innate immune cells, in particular macrophages and natural killer cells, can mediate xenograft destruction, the determinants of which are being elucidated. Chronic xenograft rejection is not well understood, but recent studies indicate that non-immunological problems, such as incompatibilities between human procoagulant and pig anticoagulant components may play an important role. Here, we give a comprehensive overview of the currently known obstacles to xenografting: immune and non-immune problems are discussed, as well as the possible strategies that are under development to overcome these hurdles. PMID- 18418355 TI - Aristolochic acid nephropathy: a worldwide problem. AB - Aristolochic acid nephropathy (AAN), a progressive renal interstitial fibrosis frequently associated with urothelial malignancies, was initially reported in a Belgian cohort of more than 100 patients after the intake of slimming pills containing a Chinese herb, Aristolochia fangchi. Although botanicals known or suspected to contain aristolochic acid (AA) were no longer permitted in many countries, several AAN cases were regularly observed all around the world. The incidence of AAN is probably much higher than initially thought, especially in Asia and the Balkans. In Asian countries, where traditional medicines are very popular, the complexity of the pharmacopoeia represents a high risk for AAN because of the frequent substitution of the botanical products by AA-containing herbs. In the Balkan regions, the exposure to AA found in flour obtained from wheat contaminated with seeds of Aristolochia clematitis could be responsible for the so-called Balkan-endemic nephropathy. Finally, despite the Food and Drug Administration's warnings concerning the safety of botanical remedies containing AA, these herbs are still sold via the Internet. PMID- 18418356 TI - Proteinuria in diabetic kidney disease: a mechanistic viewpoint. AB - Proteinuria is the hallmark of diabetic kidney disease (DKD) and is an independent risk factor for both renal disease progression, and cardiovascular disease. Although the characteristic pathological changes in DKD include thickening of the glomerular basement membrane and mesangial expansion, these changes per se do not readily explain how patients develop proteinuria. Recent advances in podocyte and glomerular endothelial cell biology have shifted our focus to also include these cells of the glomerular filtration barrier in the development of proteinuria in DKD. This review describes the pathophysiological mechanisms at a cellular level which explain why patients with DKD develop proteinuria. PMID- 18418358 TI - Nociceptin/orphanin FQ modulates motor behavior and primary motor cortex output through receptors located in substantia nigra reticulata. AB - This study was set to investigate whether motor effects of nociceptin/orphanin FQ (N/OFQ) can be related to changes in primary motor cortex output. N/OFQ injected i.c.v. biphasically modulated motor performance, low doses being facilitating and higher ones inhibitory. These effects were counteracted by the N/OFQ receptor antagonist [Nphe(1) Arg(14),Lys(15)]N/OFQ-NH(2) (UFP-101) confirming the specificity of N/OFQ action. However, UFP-101 alone facilitated motor performance, suggesting that endogenous N/OFQ inhibits motor function. N/OFQ and UFP-101 injected into the substantia nigra reticulata but not motor cortex replicated these effects, suggesting motor responses were mediated by subcortical circuits involving the basal ganglia. Intracortical microstimulation technique showed that i.c.v. N/OFQ also biphasically modulated motor cortex excitability and movement representation. Low N/OFQ doses caused a leftward shift of threshold distribution curve in the forelimb area without affecting the number of effective sites. Conversely, high N/OFQ doses increased unresponsive and reduced excitable (movement) sites in vibrissa but not forelimb area. However, increased threshold currents and rightward shift of threshold distribution curve were observed in both areas, suggesting an overall inhibitory effect on cortical motor output. UFP 101 alone evoked effects similar to low N/OFQ doses, suggesting tonic inhibitory control over forelimb movement by endogenous N/OFQ. As shown in behavioral experiments, these effects were replicated by intranigral, but not intracortical, N/OFQ or UFP-101 injections. We conclude that N/OFQ receptors located in the substantia nigra reticulata mediate N/OFQ biphasic control over motor behavior, possibly through changes of primary motor cortex output. PMID- 18418357 TI - NMDA receptors regulate nicotine-enhanced brain reward function and intravenous nicotine self-administration: role of the ventral tegmental area and central nucleus of the amygdala. AB - Nicotine is considered an important component of tobacco responsible for the smoking habit in humans. Nicotine increases glutamate-mediated transmission throughout brain reward circuitries. This action of nicotine could potentially contribute to its intrinsic rewarding and reward-enhancing properties, which motivate consumption of the drug. Here we show that the competitive N-methyl-D aspartate (NMDA) receptor antagonist LY235959 (0.5-2.5 mg per kg) abolished nicotine-enhanced brain reward function, reflected in blockade of the lowering of intracranial self-stimulation (ICSS) thresholds usually observed after experimenter-administered (0.25 mg per kg) or intravenously self-administered (0.03 mg per kg per infusion) nicotine injections. The highest LY235959 dose (5 mg per kg) tested reversed the hedonic valence of nicotine from positive to negative, reflected in nicotine-induced elevations of ICSS thresholds. LY235959 doses that reversed nicotine-induced lowering of ICSS thresholds also markedly decreased nicotine self-administration without altering responding for food reinforcement, whereas the alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid receptor antagonist NBQX had no effects on nicotine intake. In addition, nicotine self-administration upregulated NMDA receptor subunit expression in the central nucleus of the amygdala (CeA) and ventral tegmental area (VTA), suggesting important interactions between nicotine and the NMDA receptor. Furthermore, nicotine (1 microM) increased NMDA receptor-mediated excitatory postsynaptic currents in rat CeA slices, similar to its previously described effects in the VTA. Finally, infusion of LY235959 (0.1-10 ng per side) into the CeA or VTA decreased nicotine self-administration. Taken together, these data suggest that NMDA receptors, including those in the CeA and VTA, gate the magnitude and valence of the effects of nicotine on brain reward systems, thereby regulating motivation to consume the drug. PMID- 18418361 TI - Differential involvement of glutamatergic mechanisms in the cognitive and subjective effects of smoking. AB - There is growing preclinical evidence for the involvement of glutamate in the behavioral actions of nicotine. The aim of this study, was to investigate the role of N-methyl-D-aspartate (NMDA) receptors in the cognitive and subjective effects of smoking in humans. Sixty regular smokers took part in this double blind placebo controlled study, that investigated the effect of the NMDA antagonist memantine (40 mg) and the nicotinic-receptor antagonist mecamylamine (10 mg) on smoking-induced improvement in performance of a task of sustained attention and on smoking-induced changes in subjective effects and craving. Increases in subjective ratings of 'buzzed' following smoking were reversed by memantine, but not by mecamylamine. In contrast, improvement on a Rapid Visual Information Processing task by smoking was opposed by mecamylamine, but not by memantine. Smoking reduced craving for cigarettes, but neither drug altered this effect. Our results suggest that glutamatergic mechanisms may have differential involvement in the subjective and cognitive actions of smoking. Further investigations using different ligands are warranted to fully characterize the role of glutamate underlying the consequences of smoking behavior. PMID- 18418359 TI - Effects of substance P in the amygdala, ventromedial hypothalamus, and periaqueductal gray on fear-potentiated startle. AB - The neural pathways through which substance P (SP) influences fear and anxiety are poorly understood. However, the amygdala, a brain area repeatedly implicated in fear and anxiety processes, is known to contain large numbers of SP-containing neurons and SP receptors. Several studies have implicated SP neurotransmission within the amygdala in anxiety processes. In the present study, we evaluated the effects of site-specific infusions of an SP receptor antagonist, GR 82334, on conditioned fear responses using the fear-potentiated startle paradigm. GR 82334 infusion into the basolateral (BLA) or the medial (MeA) nuclei of the amygdala, but not into the central nucleus of the amygdala (CeA), dose dependently reduced fear-potentiated startle. Similar effects were obtained with GR 82334 infusion into the ventromedial nucleus of the hypothalamus (VMH), to which the MeA projects, and into the rostral dorsolateral periaqueductal gray (PAG), to which the VMH projects, but not into the deep layers of the superior colliculus/deep mesencephalic nucleus (dSC/DpMe), an output of the CeA previously shown to be important for fear-potentiated startle. Consistent with previous findings, infusion of the AMPA receptor antagonist, NBQX, into the dSC/DpMe, but not into the PAG, did disrupt fear-potentiated startle. These findings suggest that multiple outputs from the amygdala play a critical role in fear-potentiated startle and that SP plays a critical, probably modulatory role, in the MeA to VMH to PAG to the startle pathway based on these and data from others. PMID- 18418360 TI - Effects of memantine on neuronal structure and conditioned fear in the Tg2576 mouse model of Alzheimer's disease. AB - Memantine, an uncompetitive NMDA receptor antagonist used for the treatment of Alzheimer's disease (AD), has been hypothesized to have neuroprotective properties. However, the similarity of its mechanism of action to other NMDA receptor antagonists has led to concerns that it may also have neurotoxic effects. To assess both the neuroprotective and neurotoxic potential of memantine in a mouse model of AD (Tg2576 mice), we used quantitative light and electron microscopy to investigate the effects of long-term (6 months) administration of memantine (5, 10 and 20 mg/kg) on plaque deposition and neuronal morphology in the hippocampus and overlying cortex. A fear-conditioning paradigm was used to evaluate the behavioral consequences of any observed changes in structure. Administration of the two higher doses of memantine (10 and 20 mg/kg) was associated with a significant decrease in beta-amyloid (Abeta) plaque deposition, increases in synaptic density and the appearance of degenerating axons; the latter two effects were independent of genotype. Administration of the lowest dose of memantine (5 mg/kg) was associated with a significant decrease in Abeta plaque deposition and a significant increase in synaptic density, but not a significant increase in degenerating axons. However, memantine did not significantly improve behavioral deficits associated with genotype in a fear conditioning paradigm at any dose. These results suggest that chronic memantine administration may have both neuroprotective and neurotoxic effects in a mouse model of AD. PMID- 18418362 TI - Nucleus accumbens core acetylcholine is preferentially activated during acquisition of drug- vs food-reinforced behavior. AB - Acquisition of drug-reinforced behavior is accompanied by a systematic increase of release of the neurotransmitter acetylcholine (ACh) rather than dopamine, the expected prime reward neurotransmitter candidate, in the nucleus accumbens core (AcbC), with activation of both muscarinic and nicotinic ACh receptors in the AcbC by ACh volume transmission being necessary for the drug conditioning. The present findings suggest that the AcbC ACh system is preferentially activated by drug reinforcers, because (1) acquisition of food-reinforced behavior was not paralleled by activation of ACh release in the AcbC whereas acquisition of morphine-reinforced behavior, like that of cocaine or remifentanil (tested previously), was, and because (2) local intra-AcbC administration of muscarinic or nicotinic ACh receptor antagonists (atropine or mecamylamine, respectively) did not block the acquisition of food-reinforced behavior whereas acquisition of drug-reinforced behavior had been blocked. Interestingly, the speed with which a drug of abuse distributed into the AcbC and was eliminated from the AcbC determined the size of the AcbC ACh signal, with the temporally more sharply delineated drug stimulus producing a more pronounced AcbC ACh signal. The present findings suggest that muscarinic and nicotinic ACh receptors in the AcbC are preferentially involved during reward conditioning for drugs of abuse vs sweetened condensed milk as a food reinforcer. PMID- 18418363 TI - Estimates of serotonin and norepinephrine transporter inhibition in depressed patients treated with paroxetine or venlafaxine. AB - Paroxetine and venlafaxine are potent serotonin transporter (SERT) antagonists and weaker norepinephrine transporter (NET) antagonists. However, the relative magnitude of effect at each of these sites during treatment is unknown. Using a novel blood assay that estimates CNS transporter occupancy we estimated the relative SERT and NET occupancy of paroxetine and venlafaxine in human subjects to assess the relative magnitude of SERT and NET inhibition. Outpatient subjects (N=86) meeting criteria for major depression were enrolled in a multicenter, 8 week, randomized, double-blind, parallel group, antidepressant treatment study. Subjects were treated by forced-titration of paroxetine CR (12.5-75 mg/day) or venlafaxine XR (75-375 mg/day) over 8 weeks. Blood samples were collected weekly to estimate transporter inhibition. Both medications produced dose-dependent inhibition of the SERT and NET. Maximal SERT inhibition at week 8 for paroxetine and venlafaxine was 90% (SD 7) and 85% (SD 10), respectively. Maximal NET inhibition for paroxetine and venlafaxine at week 8 was 36% (SD 19) and 60% (SD 13), respectively. The adjusted mean change from baseline (mean 28.6) at week 8 LOCF in MADRS total score was -16.7 (SE 8.59) and -17.3 (SE 8.99) for the paroxetine and venlafaxine-treated patients, respectively. The magnitudes of the antidepressant effects were not significantly different from each other (95%CI 3.42, 4.54, p=0.784). The results clearly demonstrate that paroxetine and venlafaxine are potent SERT antagonists and less potent NET antagonists in vivo. NET antagonism has been posited to contribute to the antidepressant effects of these compounds. The clinical significance of the magnitude of NET antagonism by both medications remains unclear at present. PMID- 18418365 TI - The Roman high- and low-avoidance rat lines differ in the acquisition, maintenance, extinction, and reinstatement of intravenous cocaine self administration. AB - The selective breeding of Roman high- (RHA) and low-avoidance (RLA) rats for, respectively, rapid vs extremely poor acquisition of avoidant behavior in a shuttlebox has produced two phenotypes that differ in temperament traits, in mesocortical/mesolimbic dopamine system function, and in the behavioral and neurochemical responses to the acute and repeated administration of psychostimulants and opiates. The phenotypic traits of the RHA line predict higher susceptibility, compared with RLA rats, to the reinforcing properties of addictive substances like cocaine. The present study was designed to compare the acquisition, maintenance, reinstatement of drug-seeking after long-term extinction, and reacquisition of intravenous cocaine self-administration (SA) behavior in the Roman lines. Compared with RLA rats, the rates of responding during cocaine SA acquisition were higher, extinction from cocaine SA was prolonged, and drug-induced reinstatement of cocaine-seeking behavior was more robust in RHA rats. Moreover, only RHA rats reacquired extinguished lever pressing activity when a low reinforcing dose of cocaine was available. These findings are consistent with the view that subjects with genetically determined high responsiveness to the acute and chronic (ie, sensitizing) effects of psychostimulants, such as RHA rats, also display a higher propensity to self administer cocaine. Further comparative studies in the Roman lines, using SA paradigms that distinguish mere drug-taking from the compulsive and uncontrolled drug use that characterizes addiction in humans, may eventually help to characterize the relationships among genotype, temperament traits, and neurobiological mechanisms involved in the individual vulnerability to cocaine addiction. PMID- 18418364 TI - Norepinephrine transporter regulation mediates the long-term behavioral effects of the antidepressant desipramine. AB - The relationship between the ability of repeated desipramine treatment to cause downregulation of the norepinephrine transporter (NET) and produce antidepressant like effects on behavior was determined. Treatment of rats with 15 mg/kg per day desipramine reduced NET expression, measured by (3)H-nisoxetine binding and SDS PAGE/immunoblotting, in cerebral cortex and hippocampus and reduced the time of immobility in the forced-swim test. The antidepressant-like effect on forced-swim behavior was evident 2 days following discontinuation of desipramine treatment when plasma and brain levels of desipramine and its major metabolite desmethyldesipramine were not detectable. Reduced NET expression resulted in reduced norepinephrine uptake, measured in vitro, and increased noradrenergic neurotransmission, measured in vivo using microdialysis. Overall, the dose response and time-of-recovery relationships for altered NET expression matched those for production of antidepressant-like effects on behavior. The importance of increased noradrenergic neurotransmission in the persistent antidepressant like effect on behavior was confirmed by demonstrating that it was blocked by inhibition of catecholamine synthesis with alpha-methyl-p-tyrosine. The present results suggest an important role for NET regulation in the long-term behavioral effects of desipramine and are consistent with clinical data suggesting that enhanced noradrenergic neurotransmission is necessary, but not sufficient, for its antidepressant actions. Understanding the mechanisms underlying NET regulation in vivo may suggest novel targets for therapeutic intervention in the treatment of depression. PMID- 18418366 TI - Dose-occupancy study of striatal and extrastriatal dopamine D2 receptors by aripiprazole in schizophrenia with PET and [18F]fallypride. AB - Positron emission tomography (PET) and the high affinity D(2/3) radiotracer [(18)F]fallypride allow the assessment of D(2/3) receptor occupancy of antipsychotic drugs in striatal and extrastriatal brain regions. We measured regional occupancy attained across a range of clinical dosing by the partial D(2) agonist aripiprazole using these methods. Twenty-eight PET scans were acquired on the ECAT EXACT HR+ camera in 19 patients with schizophrenia or schizoaffective disorder. Daily aripiprazole doses ranged from 2 to 40 mg, with a minimum of 10 days on steady dose. Mean regional occupancies, a model-independent estimate of aripiprazole effect on pituitary binding, and PANSS ratings changes were evaluated. Occupancy levels were high across regions of interest, ranging from 71.6+/-5.5% at 2 mg/day to 96.8+/-5.3% at 40 mg/day. Occupancy levels were higher in extrastriatal than striatal regions. Pituitary measures of aripiprazole effect correlated with doses and were unrelated to prolactin levels, which remained within the normal range under medication. PANSS positive (but not negative) symptom improvement correlated with striatal but not extrastriatal occupancies. These data show, for the first time, D(2) occupancy by aripiprazole in treated patients with schizophrenia in extrastriatal as well as striatal regions, with high occupancy for all doses. We discuss possible explanations for higher extrastriatal than striatal occupancy. Correlations of ratings of clinical improvement with regional occupancy suggest that aripiprazole, as do other antipsychotics, benefits positive symptoms of schizophrenia most directly through its modulation of striatal rather than cortical or other extrastriatal dopamine activity. PMID- 18418367 TI - Asenapine increases dopamine, norepinephrine, and acetylcholine efflux in the rat medial prefrontal cortex and hippocampus. AB - Atypical antipsychotic drugs, which are more potent direct acting antagonists of brain serotonin (5-HT)(2A) than dopamine (DA) D(2) receptors, preferentially enhance DA and acetylcholine (ACh) efflux in the rat medial prefrontal cortex (mPFC) and hippocampus (HIP), compared with the nucleus accumbens (NAc). These effects may contribute to their ability, albeit limited, to improve cognitive function and negative symptoms in patients with schizophrenia. Asenapine (ASE), a new multireceptor antagonist currently in development for the treatment of schizophrenia and bipolar disorder, has complex serotonergic properties based upon relatively high affinity for multiple serotonin (5-HT) receptors, particularly 5-HT(2A) and 5-HT(2C) receptors. In the current study, the effects of ASE on DA, norepinephrine (NE), 5-HT, ACh, glutamate, and gamma-aminobutyric acid (GABA) efflux in rat mPFC, HIP, and NAc were investigated with microdialysis in awake, freely moving rats. ASE at 0.05, 0.1, and 0.5 mg/kg (s.c.), but not 0.01 mg/kg, significantly increased DA efflux in the mPFC and HIP. Only the 0.5 mg/kg dose enhanced DA efflux in the NAc. ASE, at 0.1 and 0.5 mg/kg, significantly increased ACh efflux in the mPFC, but only the 0.5 mg/kg dose of ASE increased HIP ACh efflux. ASE did not increase ACh efflux in the NAc at any of the doses tested. The effect of ASE (0.1 mg/kg) on DA and ACh efflux was blocked by pretreatment with WAY100635, a 5-HT(1A) antagonist/D(4) agonist, suggesting involvement of indirect 5-HT(1A) agonism in both the actions. ASE, at 0.1 mg/kg, increased NE, but not 5-HT, efflux in the mPFC and HIP. ASE, at 0.1 mg/kg (s.c.), had no effect on glutamate and GABA efflux in either the mPFC or NAc. These findings indicate that ASE is similar to clozapine and other atypical antipsychotic drugs in preferentially increasing the efflux of DA, NE, and ACh in the mPFC and HIP compared with the NAC, and suggests that, like these agents, it may also improve cognitive function and negative symptoms in patients with schizophrenia. PMID- 18418369 TI - Probing the mind: anesthesia and neuroimaging. AB - In 1947, a second power of anesthesia was described: "With anesthetic agents we seem to have a tool for producing and holding at will, and at little risk, different levels of consciousness--a tool that promises to be of great help in studies of mental phenomena." In 1995, anesthetic manipulation was coupled with neuroimaging, paving the way for detailed assessments of the relationship between the structure and the functioning of the brain. Anesthesia combined with neuroimaging thus provides a unique tool for investigating the neural correlates of human cognition. PMID- 18418368 TI - Magnetic resonance imaging investigation of axonal remodeling and angiogenesis after embolic stroke in sildenafil-treated rats. AB - Interaction between angiogenesis and axonal remodeling after stroke was dynamically investigated by MRI in rats with or without sildenafil treatments. Male Wistar rats were subjected to embolic stroke and treated daily with saline (n=10) or with sildenafil (n=11) initiated at 24 h and subsequently for 7 days after onset of ischemia. T(2)(*)-weighted imaging, cerebral blood flow (CBF), and diffusion tensor imaging (DTI) measurements were performed from 24 h to 6 weeks after embolization. T(2)(*) and fractional anisotropy (FA) maps detected angiogenesis and axonal remodeling after stroke, respectively, starting from 1 week in sildenafil-treated rats. Areas demarcated by MRI with enhanced angiogenesis, elevated local CBF, and augmented axonal remodeling were spatially and temporally matched, and FA values were significantly correlated with the corresponding CBF values (R=0.66, P<4 x 10(-5)) at 6 weeks after stroke. Axonal projections were reorganized along the ischemic boundary after stroke. These MRI measurements, confirmed by histology, showed that sildenafil treatment simultaneously enhanced angiogenesis and axonal remodeling, which were MRI detectable starting from 1 week after stroke in rats. The spatial and temporal consistency of MRI metrics and the correlation between FA and local CBF suggest that angiogenesis, by elevating local CBF, promotes axonal remodeling after stroke. PMID- 18418372 TI - The need for clinical pharmacology is greater than ever. PMID- 18418374 TI - Pharmacogenetic pathway analysis of irinotecan. AB - Irinotecan, a chemotherapeutic agent against various solid tumors, is a prodrug requiring activation to SN-38. Irinotecan's complex pharmacokinetics potentially allow for many genetic sources of variability. We explored relationships between pharmacokinetic pathways and polymorphisms in genes associated with irinotecan's metabolism and transport. We fitted a seven-compartment pharmacokinetic model with enterohepatic recirculation (EHR) to concentrations of irinotecan and metabolites SN-38, SN-38 glucuronide (SN-38G), and aminopentanoic acid (APC). Principal component analysis (PCA) of patient-specific parameter estimates produced measures interpretable along pathways. Nine principal components provided good characterization of the overall variation. Polymorphisms in genes UGT1A1, UGT1A7, and UGT1A9 had strong associations with a component corresponding to the irinotecan-to-SN-38 pathway and SN-38 recirculation and to a component relating to SN-38-to-SN-38G conversion and elimination of SN-38G. The component characterizing irinotecan's compartments was associated with HNF1alpha and ABCC2 polymorphisms. The exploratory analysis with PCA in this pharmacogenetic analysis was able to identify known associations and may have allowed identification of previously uncharacterized functional polymorphisms. PMID- 18418375 TI - The genomic and epidemiological dynamics of human influenza A virus. AB - The evolutionary interaction between influenza A virus and the human immune system, manifest as 'antigenic drift' of the viral haemagglutinin, is one of the best described patterns in molecular evolution. However, little is known about the genome-scale evolutionary dynamics of this pathogen. Similarly, how genomic processes relate to global influenza epidemiology, in which the A/H3N2 and A/H1N1 subtypes co-circulate, is poorly understood. Here through an analysis of 1,302 complete viral genomes sampled from temperate populations in both hemispheres, we show that the genomic evolution of influenza A virus is characterized by a complex interplay between frequent reassortment and periodic selective sweeps. The A/H3N2 and A/H1N1 subtypes exhibit different evolutionary dynamics, with diverse lineages circulating in A/H1N1, indicative of weaker antigenic drift. These results suggest a sink-source model of viral ecology in which new lineages are seeded from a persistent influenza reservoir, which we hypothesize to be located in the tropics, to sink populations in temperate regions. PMID- 18418376 TI - Haem homeostasis is regulated by the conserved and concerted functions of HRG-1 proteins. AB - Haems are metalloporphyrins that serve as prosthetic groups for various biological processes including respiration, gas sensing, xenobiotic detoxification, cell differentiation, circadian clock control, metabolic reprogramming and microRNA processing. With a few exceptions, haem is synthesized by a multistep biosynthetic pathway comprising defined intermediates that are highly conserved throughout evolution. Despite our extensive knowledge of haem biosynthesis and degradation, the cellular pathways and molecules that mediate intracellular haem trafficking are unknown. The experimental setback in identifying haem trafficking pathways has been the inability to dissociate the highly regulated cellular synthesis and degradation of haem from intracellular trafficking events. Caenorhabditis elegans and related helminths are natural haem auxotrophs that acquire environmental haem for incorporation into haemoproteins, which have vertebrate orthologues. Here we show, by exploiting this auxotrophy to identify HRG-1 proteins in C. elegans, that these proteins are essential for haem homeostasis and normal development in worms and vertebrates. Depletion of hrg-1, or its paralogue hrg-4, in worms results in the disruption of organismal haem sensing and an abnormal response to haem analogues. HRG-1 and HRG-4 are previously unknown transmembrane proteins, which reside in distinct intracellular compartments. Transient knockdown of hrg-1 in zebrafish leads to hydrocephalus, yolk tube malformations and, most strikingly, profound defects in erythropoiesis phenotypes that are fully rescued by worm HRG-1. Human and worm proteins localize together, and bind and transport haem, thus establishing an evolutionarily conserved function for HRG-1. These findings reveal conserved pathways for cellular haem trafficking in animals that define the model for eukaryotic haem transport. Thus, uncovering the mechanisms of haem transport in C. elegans may provide insights into human disorders of haem metabolism and reveal new drug targets for developing anthelminthics to combat worm infestations. PMID- 18418377 TI - Long-term haematopoietic reconstitution by Trp53-/-p16Ink4a-/-p19Arf-/- multipotent progenitors. AB - Haematopoiesis is maintained by a hierarchical system where haematopoietic stem cells (HSCs) give rise to multipotent progenitors, which in turn differentiate into all types of mature blood cells. HSCs maintain themselves for the lifetime of the organism because of their ability to self-renew. However, multipotent progenitors lack the ability to self-renew, therefore their mitotic capacity and expansion potential are limited and they are destined to eventually stop proliferating after a finite number of cell divisions. The molecular mechanisms that limit the proliferation capacity of multipotent progenitors and other more mature progenitors are not fully understood. Here we show that bone marrow cells from mice deficient in three genes genetically downstream of Bmi1--p16Ink4a, p19Arf and Trp53 (triple mutant mice; p16Ink4a and p19Arf are alternative reading frames of the same gene (also called Cdkn2a) that encode different proteins)- have an approximately 10-fold increase in cells able to reconstitute the blood long term. This increase is associated with the acquisition of long-term reconstitution capacity by cells of the phenotype c-kit+Sca-1+Flt3+CD150-CD48-Lin , which defines multipotent progenitors in wild-type mice. The pattern of triple mutant multipotent progenitor response to growth factors resembles that of wild type multipotent progenitors but not wild-type HSCs. These results demonstrate that p16Ink4a/p19Arf and Trp53 have a central role in limiting the expansion potential of multipotent progenitors. These pathways are commonly repressed in cancer, suggesting a mechanism by which early progenitor cells could gain the ability to self-renew and become malignant with further oncogenic mutations. PMID- 18418378 TI - Vascular normalization in Rgs5-deficient tumours promotes immune destruction. AB - The vasculature of solid tumours is morphologically aberrant and characterized by dilated and fragile vessels, intensive vessel sprouting and loss of hierarchical architecture. Constant vessel remodelling leads to spontaneous haemorrhages and increased interstitial fluid pressure in the tumour environment. Tumour-related angiogenesis supports tumour growth and is also a major obstacle for successful immune therapy as it prevents migration of immune effector cells into established tumour parenchyma. The molecular mechanisms for these angiogenic alterations are largely unknown. Here we identify regulator of G-protein signalling 5 (Rgs5) as a master gene responsible for the abnormal tumour vascular morphology in mice. Loss of Rgs5 results in pericyte maturation, vascular normalization and consequent marked reductions in tumour hypoxia and vessel leakiness. These vascular and intratumoral changes enhance influx of immune effector cells into tumour parenchyma and markedly prolong survival of tumour-bearing mice. This is the first demonstration, to our knowledge, of reduced tumour angiogenesis and improved immune therapeutic outcome on loss of a vascular gene function and establishes a previously unrecognized role of G-protein signalling in tumour angiogenesis. PMID- 18418379 TI - Chromatin decouples promoter threshold from dynamic range. AB - Chromatin influences gene expression by restricting access of DNA binding proteins to their cognate sites in the genome. Large-scale characterization of nucleosome positioning in Saccharomyces cerevisiae has revealed a stereotyped promoter organization in which a nucleosome-free region (NFR) is present within several hundred base pairs upstream of the translation start site. Many transcription factors bind within NFRs and nucleate chromatin remodelling events which then expose other cis-regulatory elements. However, it is not clear how transcription-factor binding and chromatin influence quantitative attributes of gene expression. Here we show that nucleosomes function largely to decouple the threshold of induction from dynamic range. With a series of variants of one promoter, we establish that the affinity of exposed binding sites is a primary determinant of the level of physiological stimulus necessary for substantial gene activation, and sites located within nucleosomal regions serve to scale expression once chromatin is remodelled. Furthermore, we find that the S. cerevisiae phosphate response (PHO) pathway exploits these promoter designs to tailor gene expression to different environmental phosphate levels. Our results suggest that the interplay of chromatin and binding-site affinity provides a mechanism for fine-tuning responses to the same cellular state. Moreover, these findings may be a starting point for more detailed models of eukaryotic transcriptional control. PMID- 18418380 TI - Free choice activates a decision circuit between frontal and parietal cortex. AB - We often face alternatives that we are free to choose between. Planning movements to select an alternative involves several areas in frontal and parietal cortex that are anatomically connected into long-range circuits. These areas must coordinate their activity to select a common movement goal, but how neural circuits make decisions remains poorly understood. Here we simultaneously record from the dorsal premotor area (PMd) in frontal cortex and the parietal reach region (PRR) in parietal cortex to investigate neural circuit mechanisms for decision making. We find that correlations in spike and local field potential (LFP) activity between these areas are greater when monkeys are freely making choices than when they are following instructions. We propose that a decision circuit featuring a sub-population of cells in frontal and parietal cortex may exchange information to coordinate activity between these areas. Cells participating in this decision circuit may influence movement choices by providing a common bias to the selection of movement goals. PMID- 18418381 TI - Oxygen-regulated degradation of fission yeast SREBP by Ofd1, a prolyl hydroxylase family member. AB - Sre1, the fission yeast sterol regulatory element binding protein, is an endoplasmic reticulum membrane-bound transcription factor that responds to changes in oxygen-dependent sterol synthesis as an indirect measure of oxygen availability. Under low oxygen, Sre1 is proteolytically cleaved and the released N-terminal transcription factor (Sre1N) activates gene expression essential for hypoxic growth. Here, we describe an oxygen-dependent mechanism for regulation of Sre1 that is independent of sterol-regulated proteolysis. Using yeast expressing only Sre1N, we show that Sre1N turnover is regulated by oxygen. Ofd1, an uncharacterized prolyl 4-hydroxylase-like 2-oxoglutarate-Fe(II) dioxygenase, accelerates Sre1N degradation in the presence of oxygen. However, unlike the prolyl 4-hydroxylases that regulate mammalian hypoxia-inducible factor, Ofd1 uses multiple domains to regulate Sre1N degradation by oxygen; the Ofd1 N-terminal dioxygenase domain is required for oxygen sensing and the Ofd1 C-terminal domain accelerates Sre1N degradation. Our data support a model whereby the Ofd1 N terminal dioxygenase domain is an oxygen sensor that regulates the activity of the C-terminal degradation domain. PMID- 18418382 TI - Histone methyltransferase Dot1 and Rad9 inhibit single-stranded DNA accumulation at DSBs and uncapped telomeres. AB - Cells respond to DNA double-strand breaks (DSBs) and uncapped telomeres by recruiting checkpoint and repair factors to the site of lesions. Single-stranded DNA (ssDNA) is an important intermediate in the repair of DSBs and is produced also at uncapped telomeres. Here, we provide evidence that binding of the checkpoint protein Rad9, through its Tudor domain, to methylated histone H3-K79 inhibits resection at DSBs and uncapped telomeres. Loss of DOT1 or mutations in RAD9 influence a Rad50-dependent nuclease, leading to more rapid accumulation of ssDNA, and faster activation of the critical checkpoint kinase, Mec1. Moreover, deletion of RAD9 or DOT1 partially bypasses the requirement for CDK1 in DSB resection. Interestingly, Dot1 contributes to checkpoint activation in response to low levels of telomere uncapping but is not essential with high levels of uncapping. We suggest that both Rad9 and histone H3 methylation allow transmission of the damage signal to checkpoint kinases, and keep resection of damaged DNA under control influencing, both positively and negatively, checkpoint cascades and contributing to a tightly controlled response to DNA damage. PMID- 18418383 TI - Novel TCF-binding sites specify transcriptional repression by Wnt signalling. AB - Both transcriptional activation and repression have essential functions in maintaining proper spatial and temporal control of gene expression. Although Wnt signalling is often associated with gene activation, we have identified several directly repressed targets of Wnt signalling in Drosophila. Here, we explore how individual Wnt target genes are specified for signal-induced activation or repression. Similar to activation, repression required binding of Armadillo (Arm) to the N terminus of TCF. However, TCF/Arm mediated repression by binding to DNA motifs that are markedly different from typical TCF-binding sites. Conversion of the novel motifs to standard TCF-binding sites reversed the mode of regulation, resulting in Wnt-mediated activation instead of repression. A mutant form of Arm defective in activation was still functional for repression, indicating that distinct domains of the protein are required for each activity. This study suggests that the sequence of TCF-binding sites allosterically regulates the TCF/Arm complex to effect either transcriptional activation or repression. PMID- 18418384 TI - Active remodelling of the TIM23 complex during translocation of preproteins into mitochondria. AB - The TIM23 (translocase of the mitochondrial inner membrane) complex mediates translocation of preproteins across and their insertion into the mitochondrial inner membrane. How the translocase mediates sorting of preproteins into the two different subcompartments is poorly understood. In particular, it is not clear whether association of two operationally defined parts of the translocase, the membrane-integrated part and the import motor, depends on the activity state of the translocase. We established conditions to in vivo trap the TIM23 complex in different translocation modes. Membrane-integrated part of the complex and import motor were always found in one complex irrespective of whether an arrested preprotein was present or not. Instead, we detected different conformations of the complex in response to the presence and, importantly, the type of preprotein being translocated. Two non-essential subunits of the complex, Tim21 and Pam17, modulate its activity in an antagonistic manner. Our data demonstrate that the TIM23 complex acts as a single structural and functional entity that is actively remodelled to sort preproteins into different mitochondrial subcompartments. PMID- 18418386 TI - Essential role of the chaperonin folding compartment in vivo. AB - The GroEL/GroES chaperonin system of Escherichia coli forms a nano-cage allowing single protein molecules to fold in isolation. However, as the chaperonin can also mediate folding independently of substrate encapsulation, it remained unclear whether the folding cage is essential in vivo. To address this question, we replaced wild-type GroEL with mutants of GroEL having either a reduced cage volume or altered charge properties of the cage wall. A stepwise reduction in cage size resulted in a gradual loss of cell viability, although the mutants bound non-native protein efficiently. Strikingly, a mild reduction in cage size increased the yield and the apparent rate of green fluorescent protein folding, consistent with the view that an effect of steric confinement can accelerate folding. As shown in vitro, the observed acceleration of folding was dependent on protein encapsulation by GroES but independent of GroES cycling regulated by the GroEL ATPase. Altering the net-negative charge of the GroEL cage wall also strongly affected chaperonin function. Based on these findings, the GroEL/GroES compartment is essential for protein folding in vivo. PMID- 18418385 TI - Cooperative activity of cdk8 and GCN5L within Mediator directs tandem phosphoacetylation of histone H3. AB - The human Mediator complex is generally required for expression of protein-coding genes. Here, we show that the GCN5L acetyltransferase stably associates with Mediator together with the TRRAP polypeptide. Yet, contrary to expectations, TRRAP/GCN5L does not associate with the transcriptionally active core Mediator but rather with Mediator that contains the cdk8 subcomplex. Consequently, this derivative 'T/G-Mediator' complex does not directly activate transcription in a reconstituted human transcription system. However, within T/G-Mediator, cdk8 phosphorylates serine-10 on histone H3, which in turn stimulates H3K14 acetylation by GCN5L within the complex. Tandem phosphoacetylation of H3 correlates with transcriptional activation, and ChIP assays demonstrate co occupancy of T/G-Mediator components at several activated genes in vivo. Moreover, cdk8 knockdown causes substantial reduction of global H3 phosphoacetylation, suggesting that T/G-Mediator is a major regulator of this H3 mark. Cooperative H3 modification provides a mechanistic basis for GCN5L association with cdk8-Mediator and also identifies a biochemical means by which cdk8 can indirectly activate gene expression. Indeed our results suggest that T/G Mediator directs early events-such as modification of chromatin templates-in transcriptional activation. PMID- 18418387 TI - Hzf regulates adipogenesis through translational control of C/EBPalpha. AB - Adipocyte differentiation requires a well-defined programme of gene expression in which the transcription factor C/EBPalpha (CCAAT/enhancer-binding protein) has a central function. Here, we show that Hzf (haematopoietic zinc-finger), a previously identified p53 transcriptional target, regulates C/EBPalpha expression. Hzf is induced during differentiation of preadipocyte cell lines, and its suppression by short hairpin RNA disrupts adipogenesis. In Hzf's absence, expression of C/EBPalpha is severely impaired because of reduced translation of its mRNA. Hzf physically interacts with the 3' untranslated region of C/EBPalpha mRNA to enhance its translation. Taken together, these findings underscore a critical role of Hzf in the adipogenesis regulatory cascade. PMID- 18418388 TI - Yeast and human Ysl2p/hMon2 interact with Gga adaptors and mediate their subcellular distribution. AB - The Gga proteins represent a family of ubiquitously expressed clathrin adaptors engaged in vesicle budding at the tubular endosomal network/trans Golgi network. Their membrane recruitment is commonly thought to involve interactions with Arf and signals in cargo through the so-called VHS domain. For yeast Gga proteins, however, partners binding to its VHS domain have remained elusive and Gga localization does not absolutely depend on Arf. Here, we demonstrate that yeast Gga recruitment relies on a network of interactions between the scaffold Ysl2p/Mon2p, the small GTPase Arl1p, and the flippase Neo1p. Deletion of either YSL2 or ARL1 causes mislocalization of Gga2p, whereas a neo1-69 mutant accumulates Gga2p on aberrant structures. Remarkably, Ysl2p directly interacts with human and yeast Ggas through the VHS domain, and binding to Gga proteins is also found for the human Ysl2p orthologue hMon2. Thus, Ysl2p represents an essential, evolutionarily conserved member of a network controlling direct binding and membrane docking of Ggas. Because activated Arl1p is part of the network that binds Gga2p, Arf and Arf-like GTPases may interact in a regulatory cascade. PMID- 18418390 TI - Characteristics and expectations of patients with erectile dysfunction: results of the SCORED study. AB - In an observational study in men with erectile dysfunction (ED) consulting a general practitioner (GP) or urologist in Belgium, demographics, ED characteristics (including erection hardness score), co-morbidities and treatment expectations were evaluated using a structured questionnaire. In total, 341 GPs and 41 urologists recruited 1492 patients. Most (74%) were untreated and 25% had ED for >3 years. Considering PDE5 inhibitors, erection hardness (89%) and maintenance (92%) were considered 'very important' by most patients. Only 18% of physicians initiated discussion about ED, despite 41% of patients having >or=3 known risk factors. The questionnaire was considered helpful by 81% of GPs and 83% of their patients. Overall, patients are under-diagnosed, and physicians are reluctant to ask about ED. A questionnaire including erection hardness score is useful to facilitate discussion about ED in general practice. Erection hardness and maintenance are more important to patients as compared to fast onset or long duration of action. PMID- 18418392 TI - Fractured penis: a review. AB - Fracture of the penis is a well-recognized clinical entity. The ideal management has evolved and repair remains largely surgical. We present the etiology and pathophysiology of this condition and outline the therapeutic options. PMID- 18418391 TI - Molecular mechanisms that could contribute to prolonged effectiveness of PDE5 inhibitors to improve erectile function. AB - Cyclic guanosine monophosphate (cGMP) in penile vascular smooth muscle cells (VSMC) plays a key role in promoting penile erection. Phosphodiesterase-5 (PDE5) in VSMC breaks down cGMP to counter this effect. Sildenafil (Viagra), vardenafil (Levitra) and tadalafil (Cialis), treatments for erectile dysfunction, inhibit PDE5 action. Many men with erectile dysfunction have improved erectile function after plasma inhibitor concentration falls below therapeutic levels. Maximum effect plus onset and duration of action of inhibitor determines its efficacy. The rate and extent of cellular drug accumulation and efflux of drug from smooth muscle cells plus persistence of drug effects in these cell impact these parameters. We propose possible molecular mechanisms that could account for prolonged action of PDE5 inhibitors including (1) persistence of biochemical effects after inhibitor is cleared from cells, and (2) retention of drug in VSMC beyond plasma clearance. PMID- 18418389 TI - Topoisomerase IIIalpha is required for normal proliferation and telomere stability in alternative lengthening of telomeres. AB - Topoisomerase (Topo) IIIalpha associates with BLM helicase, which is proposed to be important in the alternative lengthening of telomeres (ALT) pathway that allows telomere recombination in the absence of telomerase. Here, we show that human Topo IIIalpha colocalizes with telomeric proteins at ALT-associated promyelocytic bodies from ALT cells. In these cells, Topo IIIalpha immunoprecipitated with telomere binding protein (TRF) 2 and BLM and was shown to be associated with telomeric DNA by chromatin immunoprecipitation, suggesting that these proteins form a complex at telomere sequences. Topo IIIalpha depletion by small interfering RNA reduced ALT cell survival, but did not affect telomerase positive cell lines. Moreover, repression of Topo IIIalpha expression in ALT cells reduced the levels of TRF2 and BLM proteins, provoked a strong increase in the formation of anaphase bridges, induced the degradation of the G-overhang signal, and resulted in the appearance of DNA damage at telomeres. In contrast, telomere maintenance and TRF2 levels were unaffected in telomerase-positive cells. We conclude that Topo IIIalpha is an important telomere-associated factor, essential for telomere maintenance and chromosome stability in ALT cells, and speculate on its potential mechanistic function. PMID- 18418393 TI - Yohimbine enhances the effect of sildenafil on erectile process in rats. AB - Combining the centrally acting drug yohimbine with the peripheral conditioner sildenafil might be an approach to erectile dysfunction cases in which sildenafil alone failed. This work aimed to investigate the effect of yohimbine on sildenafil-induced facilitation of erectile process. Erectile responses to electrical stimulation of the cavernous nerve in anesthetized male rats were recorded. Intracavernosal pressure/systemic arterial pressure (ICP/SAP) was calculated, 1 and 5 min after intravenous administration of sildenafil, yohimbine or a combination of both. Changes in sexual arousal and copulatory performance indices were compared before and after these injections using behavioral mating experiments. It was shown that systemic administration of sildenafil produced a significant increase in ICP/SAP than control at doses >or=10 micromol kg(-1). Yohimbine alone failed to potentiate erectile responses but yohimbine (1 micromol kg(-1)) significantly potentiated the effect of sildenafil 1-10 micromol kg(-1) and 1 mmol kg(-1), 1 and 5 min after injection. Potentiation of ICP/SAP induced by their combination was greater than the sum of the effects of the corresponding doses of either drug at the same time interval. A nonsignificant additional decrease in SAP than sildenafil-induced was observed if administered with yohimbine. Addition of sildenafil to yohimbine significantly enhanced the effect of the latter on intromission frequency, intercopulatory interval and the number of ejaculations per session. It is concluded that yohimbine may enhance and prolong the effect of sildenafil on erectile process without additional hypotension. Sildenafil may enhance the central effects of yohimbine on erection; it amplifies the effect of yohimbine on male copulatory performance but not on sexual motivation. The potential beneficial effect of the combination was found to be more pronounced on the central component than on the peripheral component of the erectile process. PMID- 18418394 TI - Association study of IL2/IL21 and FcgRIIa: significant association with the IL2/IL21 region in Scandinavian coeliac disease families. AB - The first genome-wide association study performed in a UK coeliac disease (CD) case-control cohort revealed association with a linkage disequilibrium block containing the KIAA1109/Tenr/IL2/IL21 genes. Also recently, an association with a non-synonymous polymorphism in FcgammaRIIa (CD32a) was reported in CD with an unusually strong P-value. We aimed to replicate the reported associations with the single nucleotide polymorphisms rs13119723 A>G and rs6822844 G>T in the KIAA1109/Tenr/IL2/IL21 region and rs1801274 G>A in the FcgammaRIIa gene in a family sample consisting of 325 Swedish/Norwegian families using the robust transmission disequilibrium test. The family sample used in this study included 100 families with two or more children affected by CD and 225 families with one affected child. We could confirm significant association between the polymorphisms rs13119723 A>G and rs6822844 G>T located in the KIAA1109/Tenr/IL2/IL21 region and CD (P-value 0.001 and 0.002, respectively). However, we found no association with the FcgammaRIIa rs1801274 G>A polymorphism (P-value=0.3). In conclusion, our results support the KIAA1109/Tenr/IL2/IL21 region as a true CD susceptibility region. PMID- 18418395 TI - Comprehensive association study of genetic variants in the IL-1 gene family in systemic juvenile idiopathic arthritis. AB - Patients with systemic juvenile idiopathic arthritis (sJIA) have a characteristic daily spiking fever and elevated levels of inflammatory cytokines. Members of the interleukin-1 (IL-1) gene family have been implicated in various inflammatory and autoimmune diseases, and treatment with the IL-1 receptor antagonist, Anakinra, shows remarkable improvement in some patients. This work describes the most comprehensive investigation to date of the involvement of the IL-1 gene family in sJIA. A two-stage case-control association study was performed to investigate the two clusters of IL-1 family genes using a tagging single nucleotide polymorphism (SNP) approach. Genotyping data of 130 sJIA patients and 151 controls from stage 1 highlighted eight SNPs in the IL1 ligand cluster region and two SNPs in the IL1 receptor cluster region as showing a significant frequency difference between the populations. These 10 SNPs were typed in an additional 105 sJIA patients and 184 controls in stage 2. Meta-analysis of the genotypes from both stages showed that three IL1 ligand cluster SNPs (rs6712572, rs2071374 and rs1688075) and one IL1 receptor cluster SNP (rs12712122) show evidence of significant association with sJIA. These results indicate that there may be aberrant control of the activity of the IL-1 family in sJIA patients causing the increased susceptibility to the disease. PMID- 18418396 TI - Susceptibility to Coccidioides species in C57BL/6 mice is associated with expression of a truncated splice variant of Dectin-1 (Clec7a). AB - Coccidioides posadasii spherules stimulate macrophages to make cytokines via TLR 2 and Dectin-1. We used formalin-killed spherules and 1,3-beta-glucan purified from spherules to stimulate elicited peritoneal macrophages and myeloid dendritic cells (mDCs) from susceptible (C57BL/6) and resistant (DBA/2) mouse strains. DBA/2 macrophages produced more TNF-alpha and IL-6 than macrophages from C57BL/6 mice, and the amount of TNF-alpha made was dependent on both TLR2 and Dectin-1. DCs from C57BL/6 mice made more IL-10 and less IL-23p19 and IL-12p70 than did DBA/2 DC. These responses were inhibited by a monoclonal antibody to Dectin-1. DBA/2 mice expressed full-length Dectin-1, whereas C57BL/6 mice spliced out exon 3, which encodes most of the stalk. RAW cells transduced to express the full length Dectin-1 responded better to FKS than cells expressing truncated Dectin-1. We compared the isoform of Dectin-1 expressed by 34 C57BL/6 X DBA/2 recombinant inbred (BXD RI) lines with their susceptibility to Coccidioides immitis. In 25 of 34 RI lines susceptibility or resistance corresponded to short or full-length isoforms, respectively. These results suggest that alternative splicing of the Dectin-1 gene contributes to susceptibility of C57BL/6 mice to coccidioidomycosis, and affects the cytokine responses of macrophages and mDCs to spherules. PMID- 18418397 TI - Associations between the human MHC and sustained virologic response in the treatment of chronic hepatitis C virus infection. AB - The human major histocompatability complex (MHC) genes encode the human leukocyte antigens, which are important in antigen presentation and regulation of CD8+ and CD4+ T cells. Response to therapies in hepatitis C virus (HCV) infection is highly variable (30-80%) and lower response rates have been reported among African Americans (AA; approximately 30%) compared to Caucasian Americans (CA; approximately 50%) infected with genotype-1 viruses. We evaluated whether MHC gene variants were associated with response to therapy and racial differences in AA and CA sustained virologic response (SVR) rates. We genotyped alleles at 8 MHC loci: 3 class I (A, B and C) and 5 class II (DRB1, DQA1, DQB1, DPA1 and DPB1) loci in 373 individuals (179 AA and 194 CA) with genotype-1 HCV infections, who were treated with peginterferon-alpha-2a and ribavirin. We observed carriage of A(*)02 (RR=1.33(1.08-1.64); P=0.008), B(*)58 (RR=1.84(1.24-2.73); P=0.002) and DPB1(*)1701 (RR=1.57(1.09-2.26); P=0.015) to be associated with SVR after adjustment for other predictors of response. In analysis of AA and CA subgroups separately, we observed potential, though not statistically significant, differences in these MHC associations. Variation in the immunogenetic background of HCV-infected individuals might account for some observed variation in viral specific immunity and courses of disease. In this regard, future studies examining broader patient populations are warranted. PMID- 18418398 TI - Onecut transcription factor OC2 is a direct target of T-bet in type-1 T-helper cells. AB - T-box transcription factor, T-bet, has a central role in the differentiation of T helper (Th) progenitor cells to Th1 or Th2 effector cells, partly by regulating the expression of genes such as interferon-gamma (IFN-gamma). However, the direct target genes, especially those mediating the transcriptional network initiated by T-bet, are not yet fully understood. By combining chromatin immunoprecipitation from Th1 cells with human cytosine-phosphate-guanine-island array analysis, Onecut 2 (OC2), which encodes a member of the ONECUT class of transcriptional activators, was identified as a direct target gene of T-bet. OC2 is expressed in Th1 but not Th2 cells and reporter assays showed that T-bet transactivates OC2 transcription through putative T-bet half-sites locating -451 to -347 of OC2 promoter region. Moreover, we found that OC2 binds and transactivates human T-bet promoter. These results suggest that not only cell-extrinsic regulation via the IFN-gamma/STAT1 pathway, but also cell-intrinsic transcriptional positive feedback loop between T-bet and OC2 could be involved in Th1 development. PMID- 18418399 TI - Pharmacological interventions for hypertensive emergencies: a Cochrane systematic review. AB - Hypertensive emergencies occur when high blood pressure is associated with the presence of acute end-organ damage, such as hypertensive encephalopathy. There is controversy as to when and which antihypertensive drugs to use in these situations. Using a comprehensive search strategy in electronic sources, MEDLINE, EMBASE and Cochrane clinical trial register, we conducted a systematic review to look all randomized control trials (RCTs) that compare an antihypertensive drug versus placebo, no treatment or another antihypertensive drug. Fifteen RCTs (representing 869 patients) met the inclusion criteria. Two trials included a placebo arm. All studies (except one) were open-label trials. Seven drug classes were evaluated in those trials: nitrates (nine trials), angiotensin-converting enzyme inhibitors (seven), diuretics (three), calcium channel blockers (six), alpha-1 adrenergic antagonists (four), direct vasodilators (two) and dopamine agonists (one). Mortality event data were reported in seven trials. Due to insufficient data, no meta-analysis was performed for clinical outcomes. Differences in blood pressure changes between antihypertensives were minor. There is no RCT evidence demonstrating that antihypertensive drugs reduce mortality or morbidity in patients with hypertensive emergencies. Furthermore, there is insufficient RCT evidence to determine which drug or drug class is most effective in reducing mortality and morbidity. There were some minor differences in the degree of blood pressure lowering when one class of antihypertensive drug is compared to another. However, the clinical significance is unknown. RCTs are needed to assess different drug classes to determine initial and longer-term mortality and morbidity outcomes. PMID- 18418400 TI - The protective effect of KCNMB1 E65K against hypertension is restricted to blood pressure treatment with beta-blockade. PMID- 18418401 TI - Maternal educational level and risk of gestational hypertension: the Generation R Study. AB - We examined whether maternal educational level as an indicator of socioeconomic status is associated with gestational hypertension. We also examined the extent to which the effect of education is mediated by maternal substance use (that is smoking, alcohol consumption and illegal drug use), pre-existing diabetes, anthropometrics (that is height and body mass index (BMI)) and blood pressure at enrollment. This was studied in 3262 Dutch pregnant women participating in the Generation R Study, a population-based cohort study. Level of maternal education was established by questionnaire at enrollment, and categorized into high, mid high, mid-low and low. Diagnosis of gestational hypertension was retrieved from medical records using standard criteria. Odds ratios (OR) of gestational hypertension for educational levels were calculated, adjusted for potential confounders and additionally adjusted for potential mediators. Adjusted for age and gravidity, women with mid-low (OR: 1.52; 95% CI: 1.02, 2.27) and low education (OR: 1.30; 95% CI: 0.80, 2.12) had a higher risk of gestational hypertension than women with high education. Additional adjustment for substance use, pre-existing diabetes, anthropometrics and blood pressure at enrollment attenuated these ORs to 1.09 (95% CI: 0.70, 1.69) and 0.89 (95% CI: 0.50, 1.58), respectively. These attenuations were largely due to the effects of BMI and blood pressure at enrollment. Women with relatively low educational levels have a higher risk of gestational hypertension, which is largely due to higher BMI and blood pressure levels from early pregnancy. The higher risk of gestational hypertension in these women is probably caused by pre-existing hypertensive tendencies that manifested themselves during pregnancy. PMID- 18418402 TI - The risk factors for elevated blood pressure and how to address cardiovascular risk factors: a review in paediatric populations. AB - This paper examines the epidemiology of hypertension and management strategies of cardiovascular diseases (CVDs) in paediatric populations in sub-Saharan Africa and other parts of the world in the 21st century. A computerized literature search was carried out using Medline, Evidence-Based Child Health (A Cochrane Review Journal), the Cochrane Library and Cochrane Child Health Field. A manual search in the African Journal for physical, health education, recreation and dance, the South African Journal for Research in Sports, Physical Education and Recreation and a few individuals in the area were requested to send some of their recent unpublished and published reports in the field. High prevalence rates and high odds ratios for high blood pressure (BP) were recorded in children aged 5-17 years. There is a need for health ministries to improve the public health sector so as to close the gap between the rich and the poor. Although personal and parental responsibility remain crucial, it also falls on the government to help control powerful environmental factors that are leading our children to premature ill-health and mortality. Equally, health professionals are increasingly recognized that they have a significant role to play in delivering medical treatment of hypertension in children. PMID- 18418403 TI - Immunotherapy of established (pre)malignant disease by synthetic long peptide vaccines. AB - This Review deals with recent progress in the immunotherapy of established (pre)malignant disease of viral or non-viral origin by synthetic vaccines capable of inducing robust T-cell responses. The most attractive vaccine compounds are synthetic long peptides (SLP) corresponding to the sequence of tumour viral antigens or tumour-associated non-viral antigens. Crucial to induction of therapeutic T-cell immunity is the capacity of SLP to deliver specific cargo to professional antigen-presenting cells (dendritic cells (DC)). Proper DC activation then induces the therapeutic CD4+ and CD8+ T-cell responses that are associated with regression of established (pre)malignant lesions, including those induced by high-risk human papilloma virus. PMID- 18418405 TI - The WHO 2008 classification of Ph-myeloproliferative disorders: statement of the Czech MPD Working Group. PMID- 18418404 TI - Quantitative MRD monitoring identifies distinct GVL response patterns after allogeneic stem cell transplantation for chronic lymphocytic leukemia: results from the GCLLSG CLL3X trial. AB - The purpose of this study was to prospectively analyze minimal residual disease(MRD) kinetics after reduced-intensity allogeneic stem cell transplantation (allo-SCT) in high-risk chronic lymphocytic leukemia (CLL). Subjects were the first 30 consecutive patients from a prospective clinical trial, and seven pilot patients treated identically. Using real-time quantitative PCR (RQ-PCR) and/or flow-based MRD monitoring (sensitivity >or=10(-4)), five distinct patterns of MRD kinetics could be identified: patients who promptly achieved durable MRD negativity without direct evidence of graft-versus-leukemia (GVL) effects (Group 1) (n=4; no clinical relapse); patients with complete and sustained MRD response after GVL induced by immunosuppression tapering (Group 2) or donor lymphocyte infusions (Group 3) (n=18; one relapse); patients without MRD response due to lack of GVL (Group 4) (n=2; two relapses); patients with incomplete and transient MRD response to GVL (Group 5) (n=4; three relapses). In summary, this study provides a comprehensive map of possible MRD courses and their prognostic implications after T-replete allo-SCT in high-risk CLL, indicating that effective GVL activity is induced virtually in all patients who develop chronic GVHD. However, in a significant proportion of cases, this does not translate into sustained disease control due to development of secondary GVL resistance. PMID- 18418406 TI - HLA-DP as specific target for cellular immunotherapy in HLA class II-expressing B cell leukemia. AB - Mismatching for human leukocyte antigen (HLA)-DPB1 in unrelated donor hematopoietic stem cell transplantation (URD-SCT) has been associated with a decreased risk of disease relapse, indicating that HLA-DP may represent a target for graft-versus-leukemia (GVL) reactivity in HLA class II-expressing hematological malignancies. To investigate whether HLA-DP-specific T cells could mediate GVL reactivity following HLA-DPB1-mismatched URD-SCT and donor lymphocyte infusion (DLI), we analyzed the immune response in a patient with leukemic lymphoplasmacytic lymphoma responding to DLI without graft-versus-host disease. The emergence of leukemia-reactive CD4+ T cells during the clinical immune response was demonstrated by interferon-gamma (IFN-gamma) enzyme-linked immunosorbent spot(ELISPOT)analysis. Following clonal isolation of these leukemia reactive CD4+ T cells, blocking studies, panel studies and retroviral transduction experiments of both mismatched HLA-DPB1 alleles identified HLA DPB1(*)0201 and HLA-DPB1(*)0301 as the targets of this immune response. The HLA DPB1-specific CD4+ T-cell clones were capable of recognizing and lysing several HLA-DP-expressing myeloid and lymphoid hematological malignant cells. Since HLA DP expression is mainly restricted to hematopoietic cells, HLA-DP may be used as a specific target for immunotherapy following T-cell-depleted URD-SCT. Therefore, in patients with HLA class II-expressing hematological malignancies HLA-DP mismatched SCT may be preferable over fully matched SCT allowing DLI to induce a GVL effect. PMID- 18418407 TI - Disruption of the Bcr-Abl/Hsp90 protein complex: a possible mechanism to inhibit Bcr-Abl-positive human leukemic blasts by novobiocin. AB - The Bcr-Abl fusion gene encodes for the p210(Bcr-Abl) or p185(Bcr-Abl) tyrosine kinase (TK) implicated in the pathogenesis of chronic myelogenous leukemia (CML) or acute lymphoblastic leukemia, respectively. Because Bcr-Abl TK is chaperoned by Hsp90 (90 kDa heat-shock protein), we investigated the effects of novobiocin (NB), an Hsp90 C-terminal inhibitor, on the viability of the Bcr-Abl-positive human leukemia cells HL-60/Bcr-Abl and K562, the expression of Bcr-Abl protein and the interaction between Hsp90 and Bcr-Abl TK. Present studies demonstrate that NB is a potent inhibitor of the growth of Bcr-Abl-positive human leukemia cells. NB induces cytosolic accumulation of cytochrome c and activation of caspase-9 and caspase-3, triggering apoptosis of HL-60/Bcr-Abl and K562 cells. Treatment of cell lines with NB disrupts Bcr-Abl /Hsp90 and Bcr-Abl /Hsp70 interactions, resulting in a decreased amount of intracellular Bcr-Abl protein levels. Co-treatment with the proteasome inhibitor N-acetyl leucyl-leucyl norlucinal increases NB-mediated accumulation of Bcr-Abl in the detergent insoluble cellular fraction, which demonstrates that NB promotes proteasomal degradation of Bcr-Abl. Moreover, both imatinib-resistant K562/G01 and primary CML CD34(+) cells are sensitive to NB. PMID- 18418408 TI - Impact of genetic abnormalities on survival after allogeneic hematopoietic stem cell transplantation in multiple myeloma. AB - We analyzed the prognostic impact of the most frequent genetic abnormalities detected by fluorescence in situ hybridization in 101 patients with multiple myeloma, who underwent allogeneic hematopoietic stem cell transplantation (HSCT) after melphalan/fludarabine-based reduced conditioning. The incidences of abnormalities in the present analysis were as follows: del(13q14) (61%), t(11;14)(q13;q32) (14%), t(4;14)(p16.3;q32) (19%), MYC-gain gains (8q24) (21%), del(17p13) (16%) and t(14;16)(q32;q23) (5%). None of the patients had t(6;14)(p25;q32). The overall complete remission (CR) rate was 50% with no differences between the genetic abnormalities except for patients with del(17p13) who achieved less CR (7 vs 56%; P=0.001). Univariate analysis revealed a higher relapse rate in patients aged >50 years (P=0.002), patients with del(13q14) (P=0.006) and patients with del(17p13) (P=0.003). In multivariate analyses, only del(13q14) (HR: 2.34, P=0.03) and del(17p13) (HR: 2.24; P=0.04) significantly influenced the incidence of relapse, whereas for event-free survival, only age (HR 2.8; P=0.01) and del(17p13) (HR: 2.05; P=0.03) retained their negative prognostic value. These data show that del(17p13) is a negative prognostic factor for achieving CR as well as for event-free survival after HSCT. Translocation t(4;14) might be overcome by allogeneic HSCT, which will have implication for risk-adapted strategies. PMID- 18418409 TI - Amonafide L-malate is not a substrate for multidrug resistance proteins in secondary acute myeloid leukemia. PMID- 18418410 TI - CD34+CD38+CD19+ as well as CD34+CD38-CD19+ cells are leukemia-initiating cells with self-renewal capacity in human B-precursor ALL. AB - The presence of rare malignant stem cells supplying a hierarchy of malignant cells has recently been reported. In human acute myelogenous leukemia (AML), the leukemia stem cells (LSCs) have been phenotypically restricted within the CD34+CD38- fraction. To understand the origin of malignant cells in primary human B-precursor acute lymphocytic leukemia (B-ALL), we established a novel in vivo xenotransplantation model. Purified CD34+CD38+CD19+, CD34+CD38-CD19+ and CD34+CD38-CD19- bone marrow (BM) or peripheral blood (PB) cells from three pediatric B-ALL patients were intravenously injected into sublethally irradiated newborn NOD/SCID/IL2rgamma(null) mice. We found that both CD34+CD38+CD19+ and CD34+CD38-CD19+ cells initiate B-ALL in primary recipients, whereas the recipients of CD34+CD38-CD10-CD19- cells showed normal human hematopoietic repopulation. The extent of leukemic infiltration into the spleen, liver and kidney was similar between the recipients transplanted with CD34+CD38+CD19+ cells and those transplanted with CD34+CD38-CD19+ cells. In each of the three cases studied, transplantation of CD34+CD38+CD19+ cells resulted in the development of B-ALL in secondary recipients, demonstrating self-renewal capacity. The identification of CD34+CD38+CD19+ self-renewing B-ALL cells proposes a hierarchy of leukemia-initiating cells (LICs) distinct from that of AML. Recapitulation of patient B-ALL in NOD/SCID/IL2rgamma(null) recipients provides a powerful tool for directly studying leukemogenesis and for developing therapeutic strategies. PMID- 18418411 TI - An IFN-associated cytotoxic cellular immune response against viral, self-, or tumor antigens is a common pathogenetic feature in "interface dermatitis". AB - The term "interface dermatitis" (ID) involves a specific histological inflammatory pattern that is characterized by a cytotoxic lymphocytic infiltration and a hydropic degeneration of the basal epidermal layer. ID is typically seen in autoimmune skin disorders such as lichen planus (LP), cutaneous lupus erythematosus (CLE), and may also appear during immune reactions against drugs, viruses, and tumors. Recent studies have shown that the type-I IFN system is involved in cutaneous autoimmune diseases characterized by ID. IFNs induce the expression of proinflammatory cytokines and chemokines, which support the cellular immune response. The role of IFNs in ID is supported by a close morphological association between the expression pattern of IFN-inducible proteins and the distribution of CXCR3+ lymphocytes. The IFN-inducible chemokine CXCL10 is expressed in exactly those areas where cytotoxic lymphocytes invade the basal epidermis and cause keratinocyte death. A similar picture can be found in early herpes simplex viral skin lesions and viral warts, but also in "lichenoid" actinic keratosis and invasive squamous cell carcinoma. These data suggest that ID morphologically reflects a common IFN-driven cytotoxic attack affecting the basal keratinocytes under different conditions, which is important for antiviral and antitumor immune response, but is inappropriately activated in autoimmune skin disorders. PMID- 18418412 TI - Global assessment of psoriasis severity and change from photographs: a valid and consistent method. AB - Five raters tested the validity and consistency of global assessments of severity and change from standardized photographs in 30 consecutive patients with plaque psoriasis. The main outcome measures were physician global assessment (PGA) scores for change between baseline and follow-up visits ("dynamic PGA") and for severity at the baseline visit ("static PGA"). These photographic evaluation scores were compared with in-person clinical ratings. Panel ratings were obtained using the mean of the five raters' independent evaluations from photographs. Validity and consistency were assessed with intra-class coefficients (ICCs; 95% confidence interval). Intra-rater and intra-panel consistencies for photographic dynamic PGA scores were 0.85 (0.74-0.92) and 0.95 (0.92-0.99), respectively. As an evaluation of validity, agreement between photographic and clinical static PGA scores was 0.87 (0.75-0.93). We concluded that global assessment of psoriasis severity and change from photographs by a panel of experts was accurate and consistent. The generalizability of the results requires further studies. The intrinsic limitations of photographic assessment of individual characteristics such as plaque thickness and their effect on global photographic assessment should be further evaluated. PMID- 18418413 TI - Gene therapy progress and prospects cancer: oncolytic viruses. AB - The past 2 years have seen several major advances in oncolytic virotherapy. Studies on the interaction between viruses, immune responses and tumor microenvironment have provided important insight, while clinical trials have shown promise. This review summarizes key findings in this field over the past 2 years, and provides directions for future success. PMID- 18418414 TI - AAV vectors for RNA-based modulation of gene expression. AB - At the post-transcriptional level, gene expression is largely regulated through a network of molecular machines that regulate pre-mRNA maturation integrity, transport, translation and degradation. These processes are based on the formation of nucleoprotein complexes and require the recognition of sequence motifs on the RNA. By masking these targets with complementary RNA sequences forming Watson-Crick base pairing, it is possible to efficiently and specifically impact on the cell phenotype, or to compensate the deleterious effect of mutations. Here we review how the adeno-associated virus technology is being exploited for expressing non-coding RNAs in tissues such as the brain, muscle or liver, in functional genomic studies as well as for the development of novel therapeutic strategies. PMID- 18418415 TI - Clinical gene therapy using recombinant adeno-associated virus vectors. AB - Recombinant adeno-associated virus (rAAV) vectors possess a number of properties that may make them suitable for clinical gene therapy, including being based upon a virus for which there is no known pathology and a natural propensity to persist in human cells. Wild-type adeno-associated viruses (AAVs) are now known to be very diverse and ubiquitous in humans and nonhuman primates, which adds to the degree of confidence one may place in the natural history of AAV, namely that it has never been associated with any human tumors or other acute pathology, other than sporadic reports of having been isolated from spontaneously aborted fetuses. On the basis of this understanding of AAV biology and a wide range of preclinical studies in mice, rabbits, dogs and nonhuman primates, a growing number of clinical trials have been undertaken with this class of vectors. Altogether, over 40 clinical trials have now been approved. Although all previous trials were undertaken using AAV serotype 2 vectors, at least two current trials utilize AAV2 vector genomes cross-packaged or pseudotyped into AAV1 capsids, which appear to mediate more efficient gene delivery to muscle. The explosion of capsid isolates available for use as vectors to over 120 has now provided the potential to broaden the application of AAV-based gene therapy to other cell types. PMID- 18418416 TI - Oral IL-10 gene delivery in a microsphere-based formulation for local transfection and therapeutic efficacy in inflammatory bowel disease. AB - The objective of this study was to examine the potential of oral interleukin-10 (IL-10) gene therapy for the treatment of inflammatory bowel disease (IBD). Nanoparticles-in-microsphere oral system (NiMOS) was formulated with murine IL-10 expressing plasmid DNA in type-B gelatin nanoparticles, which were further encapsulated in poly(epsilon-caprolactone) microsphere matrix. Upon oral administration in an acute colitis model, IL-10 expression in the large intestine was measured by quantitative real-time PCR and ELISA. The locally expressed IL-10 was able to suppress the levels of proinflammatory cytokines, such as IFN-gamma, TNF-alpha, IL-1alpha, IL-1beta and IL-12, as well as certain chemokines. The therapeutic benefits of transfected IL-10 were further demonstrated by an increase in body weight, favorable clinical activity score, restoration in colon length and weight, and suppression of inflammatory response as assessed by tissue histological analysis and myeloperoxidase activity. The results of this study provide highly encouraging evidence of oral gene delivery and transfection and potential utility in IBD therapy. PMID- 18418417 TI - AAV-mediated gene therapy for retinal disorders: from mouse to man. AB - A wide range of retinal disorders can potentially be treated using viral vector mediated gene therapy. The most widely used vectors for ocular gene delivery are based on adeno-associated virus (AAV), because they elicit minimal immune responses and mediate long-term transgene expression in a variety of retinal cell types. Proof-of-concept experiments have demonstrated the efficacy of AAV mediated transgene delivery in a number of animal models of inherited and acquired retinal disorders. Following extensive preclinical evaluation in large animal models, gene therapy for one form of inherited retinal degeneration due to RPE65 deficiency is now being tested in three concurrent clinical trials. Here, we review different approaches for treating inherited retinal degenerations and more common acquired retinal disorders using AAV-based vectors. PMID- 18418418 TI - Manufacturing and characterizing AAV-based vectors for use in clinical studies. AB - Recombinant adeno-associated virus (AAV)-based vectors expressing therapeutic gene products have shown great promise for human gene therapy. A major challenge for translation of promising research to clinical development is the manufacture and certification of AAV vectors for clinical use. This review summarizes relevant aspects of current Good Manufacturing Practice, focusing on considerations and challenges specific for recombinant AAV. PMID- 18418419 TI - In vivo transfer of plasmid from food-grade transiting lactococci to murine epithelial cells. AB - We recently demonstrated that noninvasive food-grade Lactococcus lactis (L. lactis) can deliver eukaryotic expression plasmid in mammalian cells in vitro. Here, we evaluated, in vivo, whether a eukaryotic expression plasmid carried by lactococci can translocate to the epithelial cells of the intestinal membrane. The strain LL(pLIG:BLG1) carrying one plasmid containing a eukaryotic expression cassette encoding beta-lactoglobulin (BLG), a major allergen of cow's milk, was orally administered by gavage to mice. BLG cDNA was detected in the epithelial membrane of the small intestine of 40% of the mice and BLG was produced in 53% of the mice. Oral administration of LL(pLIG:BLG1) induced a low and transitory Th1 type immune response counteracting a Th2 response in case of further sensitization. We demonstrated for the first time the transfer of a functional plasmid to the epithelial membrane of the small intestine in mice by noninvasive food-grade lactococci. PMID- 18418420 TI - A novel magnetic approach to enhance the efficacy of cell-based gene therapies. AB - Attempts have been made to use various forms of cellular vectors to deliver therapeutic genes to diseased tissues like malignant tumours. However, this approach has proved problematic due to the poor uptake of these vectors by the target tissue. We have devised a novel way of using magnetic nanoparticles (MNPs) to enhance the uptake of such 'therapeutically armed' cells by tumours. Monocytes naturally migrate from the bloodstream into tumours, so attempts have been made to use them to deliver therapeutic genes to these sites. However, transfected monocytes injected systemically fail to infiltrate tumours in large numbers. Using a new in vitro assay for assessing monocyte extravasation, we show that the ability of transfected human monocytes to migrate across a human endothelial cell layer into a 3D tumour spheroid is markedly increased when cells are pre-loaded with MNPs and a magnetic force is applied close to the spheroid. Furthermore, systemic administration of such 'magnetic' monocytes to mice bearing solid tumours led to a marked increase in their extravasation into the tumour in the presence of an external magnet. This new magnetic targeting approach could be used to increase the targeting, and thus the efficacy, of many cell-based gene therapies in vivo. PMID- 18418421 TI - Both enalapril and losartan attenuate sarcolemmal Na+-K+-ATPase remodeling in failing rat heart due to myocardial infarction. AB - To investigate the mechanisms underlying the depressed sarcolemmal (SL) Na(+) K(+)-ATPase activity in congestive heart failure (CHF), different isoforms and gene expression of Na(+)-K(+)-ATPase were examined in the failing left ventricle (LV) at 8 weeks after myocardial infarction (MI). In view of the increased activity of renin-angiotensin system (RAS) in CHF, these parameters were also studied after 5 weeks of treatment with enalapril (10 mg x kg-1 x day-1), an angiotensin-converting enzyme inhibitor, and losartan (20 mg.kg-1.day-1), an angiotensin II type 1 receptor antagonist, starting at 3 weeks after the coronary ligation in rats. The infarcted animals showed LV dysfunction and depressed SL Na(+)-K(+)-ATPase activity. Protein content and mRNA levels for Na(+)-K(+)-ATPase alpha2 isoform were decreased whereas those for Na(+)-K(+)-ATPase alpha3 isoform were increased in the failing LV. On the other hand, no significant changes were observed in protein content or mRNA levels for Na(+)-K(+)-ATPase alpha1 and beta1 isoforms. The treatment of infarcted animals with enalapril or losartan improved LV function and attenuated the depression in Na(+)-K(+)-ATPase alpha2 isoform as well as the increase in alpha3 isoform, at both the protein and mRNA levels; however, combination therapy with enalapril and losartan did not produce any additive effects. These results provide further evidence that CHF due to MI is associated with remodeling of SL membrane and suggest that the blockade of RAS plays an important role in preventing these alterations in the failing heart. PMID- 18418422 TI - Dose-dependent anticonvulsive effect of ethanol on lindane-induced seizures in rats. AB - Chronic ethanol consumption is a major risk factor for epilepsy, and seizures frequently occur during the withdrawal period. The aim of our study was to investigate effects of ethanol on lindane-induced seizures in rats. Male Wistar rats were injected i.p. with one of the following 5 treatments: (i) saline, (ii) dimethylsulfoxide, (iii) lindane (8 mg/kg) (L), (iv) ethanol in doses of 0.5 g/kg (E(0.5)), 1 g/kg (E(1)), and 2 g/kg (E(2)), and (v) groups that received ethanol 30 min before lindane (LE(0.5), LE(1), and LE(2)). Behavioral changes were described by using a descriptive scale as follows: 0, no response; 1, head nodding, lower jaw twitching; 2, myoclonic body jerks, bilateral forelimb clonus; 3, generalized tonic-clonic convulsions; 4, status epilepticus. The incidence of convulsions in the LE(2) group was significantly lower than the incidence in the L (p < 0.01) and LE(0.5) groups (p < 0.05). The median grade of convulsive behavior was significantly lower in the LE(2) (p < 0.01) and LE(1) groups (p < 0.05) compared with the L group. Latencies to the first seizure response were not significantly different among groups. ED50 of ethanol was 1.40 (1.19-1.65). Our findings suggest that ethanol decreased severity and incidence of lindane-induced seizures in a dose-dependent manner. PMID- 18418423 TI - Cholesterol-induced stimulation of platelet aggregation is prevented by a hempseed-enriched diet. AB - Hypercholesterolemia indirectly increases the risk for myocardial infarction by enhancing the ability of platelets to aggregate. Diets enriched with polyunsaturated fatty acids (PUFAs) have been shown to reduce the detrimental effects of cholesterol on platelet aggregation. This study investigated whether dietary hempseed, a rich source of PUFAs, inhibits platelet aggregation under normal and hypercholesterolemic conditions. Male New Zealand white rabbits were fed one of 6 dietary interventions: regular control diet (RG); control diet + 10% hempseed (HP); control diet + 10% partially delipidated hempseed (DHP); control diet + 0.5% cholesterol (OL); control diet + 0.5% cholesterol + 10% hempseed (OLHP); control diet + 5% coconut oil (CO). After 8 weeks, blood was collected to measure ADP- and collagen-induced platelet aggregation and plasma levels of fatty acids, cholesterol, and triglycerides. The hempseed-fed animals (HP and OLHP) displayed elevated plasma levels of PUFAs and a prominent enhancement in 18:3n-6 (gamma-linolenic acid, GLA) levels, a unique PUFA found in hempseed. The cholesterol-supplemented groups (OL and OLHP) had significantly elevated plasma levels of cholesterol and triglycerides, but platelet aggregation was significantly augmented only in the OL group. The addition of hempseed to this diet (OLHP) normalized aggregation. The direct addition of GLA to the OL platelet samples blocked the cholesterol-induced stimulation of platelet aggregation. The results of this study demonstrate that when hempseed is added to a cholesterol enriched diet, cholesterol-induced platelet aggregation returns to control levels. This normalization is not due to a reduction in plasma cholesterol levels, but may be partly due to increased levels of plasma GLA. PMID- 18418424 TI - Properties of thiamine transport in isolated perfused hearts of chronically alcoholic guinea pigs. AB - The aim of this study was to determine the mechanism of transport of (14)C thiamine in the hearts of healthy (nonalcoholic) and chronically alcoholic guinea pigs. We used the single-pass, paired-tracer dilution method on isolated and retrogradely perfused guinea pig hearts. The maximal cellular uptake (U(max)) and total cellular uptake (U(tot)) of (14)C-thiamine were determined under control conditions and under influence of possible modifiers. We tested how the presence of unlabeled thiamine, metabolic inhibitors, or absence of sodium ions influence the transport of (14)C-thiamine. The results of our experiments show that the transport of (14)C-thiamine is specific and energy-dependent and that its properties are significantly changed under the influence of chronic alcoholism. The latter effect occurs by increase in both U(max) and U(tot), as a manifestation of a compensatory mechanism in thiamine deficiency. PMID- 18418425 TI - Proapoptotic and antiapoptotic effects of hyperglycemia: role of insulin signaling. AB - Glucose toxicity is an important initiator of cardiovascular disease, contributing to the development of insulin resistance, impaired contractile function, abnormal energy metabolism, cardiomyocyte and endothelial cell death, coronary heart disease, and heart failure. High blood glucose can, however, paradoxically protect the heart against a variety of insults, including ischemia, hypoxia, and calcium overload. To provide information on the underlying basis of these divergent actions of high glucose, the present study examined the hypothesis that the adverse effects of high glucose are linked to impaired insulin signaling, leading to a reduction in the levels of cytoprotective factors, and that the beneficial effects of high glucose occur in the absence of insulin and result in an improvement in Akt signaling. This hypothesis was evaluated by using an in vitro cardiomyocyte model that is amenable to manipulations in glucose and insulin. Prolonged exposure of the isolated neonatal cardiomyocyte to medium containing insulin and high glucose led to increased susceptibility to angiotensin II-mediated apoptosis, an effect associated with reduced levels of phospho-Akt and an increased Bax/Bcl-2 ratio. By contrast, exposure to high glucose levels in the absence of insulin rendered the cardiomyocyte resistant to angiotensin II-mediated apoptosis. Because the beneficial effects of high glucose were associated with elevations in phospho-Akt and Bcl-2 content, the cardioprotective activity of high glucose resembles the actions of insulin. Hence, the activation state of Akt is largely determined by the activity of insulin and other growth factors. Because high glucose diminishes insulin signaling, it reduces phospho-Akt levels and renders the cell susceptible to damaging insults. In the absence of insulin, however, the natural activity of high glucose is unmasked. As a result, Akt signaling is increased and the cell is rendered resistant to cell death. PMID- 18418426 TI - Correlation between electrocorticographic and motor phenomena in lindane-induced experimental epilepsy in rats. AB - We report a study on the relation between open-field behavior and electroencephalographic (EEG) changes during lindane-induced seizures in 2-month old adult male Wistar rats. For chronic EEG recordings and power spectra analysis, 3 electrodes were implanted into the skull. Three groups of animals, (i) saline-injected control (n = 6), (ii) DMSO-treated (n = 6), and (iii) lindane intraperitoneally administered: L(4) (4 mg/kg, n = 10), L(6) (6 mg/kg, n = 11), and L(8) (8 mg/kg, n = 11), were observed for 30 min for the occurrence of convulsive behavior. It was assessed by incidence of motor seizures, and seizure severity grade was determined by a descriptive rating scale (0, no response; 1, head nodding, lower jaw twitching; 2, myoclonic body jerks, bilateral forelimb clonus with full rearing; 3, progression to generalized clonic convulsions followed by tonic extension of fore and hind limbs and tail; 4, status epilepticus). EEG signal and spectral analyses were suitable to describe the dynamics of complex behavioral responses. Incidence and severity of epileptic manifestations, recorded as high voltage spike-wave complexes, polyspikes, sleep like patterns in EEG, and power spectra changes, were greater in lindane-treated groups in a dose-dependent manner compared with control or DMSO-treated groups. Our results suggest good correlation between lindane-induced epileptiform activity and behavioral changes. PMID- 18418427 TI - Antiplatelet therapy mitigates cardiac remodeling and dysfunction in congestive heart failure due to myocardial infarction. AB - Antiplatelet agents such as sarpogrelate (SAR), a 5-hydroxytryptamine antagonist, and cilostazol (CIL), a phosphodiesterase-III inhibitor, are used in the management of peripheral vascular disease. In this study, we tested the hypothesis that both SAR and CIL prevent cardiac remodeling and improve cardiac function in congestive heart failure (CHF) due to myocardial infarction (MI). Post-MI rats (3 weeks after the occlusion of coronary artery) received either vehicle (MI+V, n = 36), SAR (MI+SAR; 5 mg xc kg(-1) x day(-1), n = 35) or CIL (MI+CIL; 5 mg x kg(-1) x day(-1), n = 34) from day 21 to day 56. Sham-operated rats (n = 29) served as controls. Electrocardiographic, echocardiographic, and hemodynamic parameters were measured on day 56. Treatment of infarcted animals with SAR or CIL significantly improved the left ventricular (LV) dimensions, LV fractional shortening, cardiac output, stroke volume, mean arterial pressure, LV diastolic function, and LV systolic pressure, as well as rates of LV pressure development and pressure decay. Although cardiac hypertrophy was reduced, both SAR and CIL had no effect on infarct size or MI-associated QTc prolongation. However, SAR decreased whereas CIL increased the incidence of ventricular arrhythmias and the mean number of episodes in infarcted animals. Mortality during the treatment period was decreased by 17% with SAR and increased by 10% with CIL, but these changes were not significant statistically. The data in this study suggest that both SAR and CIL prevent cardiac remodeling and improve cardiac function in MI-induced CHF; however, CIL unlike SAR increased the incidence of arrhythmias and adversely affected patient mortality. PMID- 18418428 TI - Reduced levels of cyclic AMP contribute to the enhanced oxidative stress in vascular smooth muscle cells from spontaneously hypertensive rats. AB - We have earlier shown that aortic vascular smooth muscle cells (VSMC) from 12 week-old spontaneously hypertensive rats (SHR) exhibited enhanced production of superoxide anion (O(2)(-)) compared with Wistar-Kyoto (WKY) rats. This production was attenuated to control levels by losartan, an angiotensin II (Ang II) AT(1) receptor antagonist, suggesting that the AT(1) receptor is implicated in enhanced oxidative stress in SHR. Since AT(1) receptor activation signals via adenylyl cyclase inhibition and decreases cAMP levels, it is possible that AT(1) receptor mediated decreased levels of cAMP contribute to the enhanced production of O(2)( ) in SHR. The present study was undertaken to investigate this possibility. The basal adenylyl cyclase activity as well as isoproterenol and forskolin-mediated stimulation of adenylyl cyclase was significantly attenuated in VSMC from 12-week old SHR compared with those from WKY rats, whereas Ang II-mediated inhibition of adenylyl cyclase was significantly enhanced by about 70%, resulting in decreased levels of cAMP in SHR. NADPH oxidase activity and the levels of O2- were significantly higher (about 120% and 200%, respectively) in VSMC from SHR than from WKY rats. In addition, the levels of p47(phox) and Nox4 proteins, subunits of NADPH oxidase, were significantly augmented about 35%-40% in VSMC from SHR compared with those from WKY rats. Treatment of VSMC from SHR with 8Br-cAMP, as well as with cAMP-elevating agents such as isoproterenol and forskolin, restored to control WKY levels the enhanced activity of NADPH oxidase and the enhanced levels of O(2)(-), p47(phox), and Nox4. Furthermore, in the VSMC A10 cell line, 8Br-cAMP also restored the Ang II-evoked enhanced production of O(2)(-), NADPH oxidase activity, and enhanced levels of p47(phox) and Nox4 proteins to control levels. These data suggest that decreased levels of cAMP in SHR may contribute to the enhanced oxidative stress in SHR and that increasing the levels of cAMP may have a protective effect in reducing oxidative stress and thereby improve vascular function. PMID- 18418429 TI - Lipid and lipoprotein profile in women with polycystic ovary syndrome. AB - Polycystic ovary syndrome (PCOS) is a common endocrine disorder characterized by obesity-related risk factors for cardiovascular disease. The objective of our study was to determine values of key lipid and lipoprotein fractions in PCOS, and their possible relation to insulin resistance. A total of 75 women with PCOS (aged 23.1 +/- 5.1 years, BMI 24.9 +/- 4.7 kg/m(2)), and 56 age- and BMI-matched controls were investigated. In all subjects, basal glucose, cholesterol (total, HDL, and LDL), oxidized LDL (OxLDL), triglycerides, apolipoprotein (apo)A1, apoB, and apoE, nonesterified fatty acids, insulin, testosterone, sex hormone-binding globulin, homeostasis model assessment (HOMA) index, and free androgen index were determined in the follicular phase of the cycle. PCOS patients compared with controls had increased indices of insulin resistance, basal insulin (p < 0.001), and HOMA index (p < 0.001), and worsened insulin resistance-related dyslipidemia with decreased HDL cholesterol (p < 0.01), elevated triglycerides (p = 0.010), and pronounced LDL oxidation (p < 0.001). In conclusion, characteristic dyslipidemia of insulin resistance and unfavorable proatherogenic lipoprotein ratios were present only in women with PCOS and not in controls. Elevated OxLDL and the relation of apoE and nonesterified fatty acids with insulin resistance suggest that women with PCOS are at increased risk for premature atherosclerosis. PMID- 18418430 TI - Inflammatory cytokines and malnutrition as related to risk for cardiovascular disease in hemodialysis patients. AB - Malnutrition and inflammation are associated with end-stage renal disease (ESRD). Interleukin (IL)-6 and tumor necrosis factor alpha (TNF-alpha) powerfully predict death from cardiovascular disease. The aim of our study was to establish an association between markers of inflammation and parameters of malnutrition in patients on hemodialysis. The study population consisted of 42 hemodialysis patients with different parameters of malnutrition. Blood samples were taken after an overnight fast, and plasma lipid profiles (total cholesterol, LDL cholesterol, HDL cholesterol, and triglycerides) were measured by using conventional enzymatic methods. Serum urea and creatinine levels were also measured by routine procedures. Plasma high-sensitivity C-reactive protein level (hs-CRP), TNF-alpha, and IL-6 were measured by enzyme-linked immunosorbent assay (ELISA). Standard Doppler echo examinations were used to determine plaque on carotid arteries, and end-diastolic diameter (EDD) and ejection fraction (EF) were measured by echocardiography. Malnourished patients exhibited significantly greater evidence of cardiovascular disease and carotid plaques. Factor (principal component) analysis indicated 6 latent factors with 67.5% of the variance explained within all investigated parameters. Cluster analysis was used to distinguish the inflammatory markers and the nutritional markers from other parameters and to visualize similarities between variables. In summary, this cross-sectional study in hemodialysis patients found a high prevalence of malnutrition, inflammation, carotid plaques, and cardiovascular disease. Malnourished dialysis patients are more often found with cardiovascular disease and carotid plaques. In addition, these patients have higher levels of inflammatory cytokines, which may partly explain the elevated risk for atherosclerotic vascular disease. PMID- 18418431 TI - Effects of long-term losartan and L-arginine treatment on haemodynamics, glomerular filtration, and SOD activity in spontaneously hypertensive rats. AB - Recently, it has been reported that losartan, an angiotensin II receptor (ATR) antagonist, depresses the angiotensin II-induced production of superoxide radicals. Also, in spontaneously hypertensive rats (SHR) endothelial dysfunction is associated with decreased nitric oxide (NO) synthesis. In this study, we examined the effects of long-term ATR blockade and L-arginine supplementation on the haemodynamic parameters, glomerular filtration, and oxidative status in SHR. Adult male SHR were treated with losartan (10 mg/kg) and with the NO donor L arginine (2 g/kg) for 4 weeks. The animals were divided into the following experimental groups: control (n = 7), L-arginine (n = 7), losartan (n = 7), and L arginine + losartan (n = 7). Mean arterial pressure (MAP), regional blood flow, urea clearance, and activity of superoxide dismutase (SOD) were measured at the end of treatment. MAP was significantly reduced in the losartan group compared with the control group (133.3 +/- 7.3 vs. 161.5 +/- 14.5 mm Hg). Aortic blood flow was significantly higher and aortic vascular resistance was significantly lower in all treated groups than in the control. Urea clearance rose significantly in the L-arginine + losartan group compared with control (393.27 +/ 37.58 vs. 218.68 +/- 42.03 microL x min(-1) x 100 g(-1)) as did the activity of SOD (1668.97 +/- 244.57 vs. 1083.18 +/- 169.96 U/g Hb). Our results suggest that the antihypertensive effect of losartan and L-arginine in SHR is not primarily mediated by increased SOD activity. Also, combined treatment with ATR blockade and L-arginine supplementation has a beneficial effect on renal function that is, at least in part, mediated by increased SOD activity in SHR. PMID- 18418432 TI - Genetic possibilities for altering sunflower oil quality to obtain novel oils. AB - The sunflower is one of the four most important oilseed crops in the world, and the nutritional quality of its edible oil ranks among the best vegetable oils in cultivation. Typically up to 90% of the fatty acids in conventional sunflower oil are unsaturated, namely oleic (C 18:1, 16%-19%) and linoleic (C 18:2, 68%-72%) fatty acids. Palmitic (C 16:0, 6%), stearic (C 18:0, 5%), and minor amounts of myristic (C 14:0), myristoleic (C 14:1), palmitoleic (C 16:1), arachidic (C 20:0), behenic (C 22:0), and other fatty acids account for the remaining 10%. Advances in modern genetics, most importantly induced mutations, have altered the fatty acid composition of sunflower oil to a significant extent. Treating sunflower seeds with gamma- and X-rays has produced mutants with 25%-30% palmitic acid. Sunflower seed treatment with X-rays has also resulted in mutants having 30% palmitoleic acid, while treatments with mutagenic sodium azide have produced seeds containing 35% stearic acid. The most important mutations have been obtained by treatment with dimethyl sulfate, which produced genotypes with more than 90% oleic acid. Mutants have also been obtained that have a high linoleic acid content (>80%) by treating seeds with X-rays and ethyl methanesulfonate. Of the vitamin E family of compounds, sunflower oil is known to predominantly contain alpha-tocopherol (>90%). Spontaneous mutations controlled by recessive genes have been discovered that significantly alter tocopherol forms and levels. The genes in question are tph(1) (50% alpha- and 50% beta-tocopherol), tph(2) (0% 5% alpha- and 95%-100% gamma-tocopherol), and tph(1)tph(2) (8%-40% alpha-, 0%-25% beta-, 25%-84% gamma-, and 8%-50% delta-tocopherol). The existence of (mutant) genes for increased levels of individual fatty acids and for different forms and levels of tocopherol enables the development of sunflower hybrids with different oil quality. The greatest progress has been made in developing high-oleic hybrids (>90% oleic acid). There has been considerable work done recently on the development of high-oleic hybrids with altered tocopherol levels, the oil of which will have 10-20 times greater oxidative stability than that of conventional sunflower oil. While sunflower breeders work on developing hybrids with altered oil quality, medical scientists in general and nutritionists in particular will determine the parameters for the use of these novel types of oil that can improve human nutrition and be used in the prevention of cardiovascular diseases. PMID- 18418433 TI - Oral glucose tolerance test reduces arterial baroreflex sensitivity in older adults. AB - Although postprandial decreases in blood pressure are a common cause of syncope in the older adult population, the postprandial effects of the oral glucose tolerance test on blood pressure and the arterial baroreflex remain poorly characterized in older adults. Therefore, arterial blood pressure and the arterial baroreflex were studied in 19 healthy older adults (mean age 71.7 +/- 1.1 years) who were given a standardized oral glucose load (75 g) or an isovolumetric sham drink during 2 separate sessions. All measures were taken for 120 min after treatment. Baroreflex function was assessed by using the spontaneous baroreflex method (baroreflex sensitivity, BRS). Subjects demonstrated a decrease in BRS after oral glucose that was not seen in the placebo session (two-way analysis of variance, p = 0.04). There was no significant change in systolic, mean, or diastolic blood pressure; together with a drop in BRS, this resulted in a significant tachycardia post glucose (two-way analysis of variance, p < 0.001). We conclude that healthy older adults can successfully maintain blood pressure during an oral glucose tolerance test despite a decrease in arterial BRS. Decreased BRS resulted in a tachycardic response to glucose. PMID- 18418434 TI - Half-logistic time constant: a more reliable lusitropic index than monoexponential time constant regardless of temperature in canine left ventricle. AB - Temperature changes influence cardiac diastolic function. The monoexponential time constant (tauE), which is a conventional lusitropic index of the rate of left ventricular (LV) pressure fall, increases with cooling and decreases with warming. We have proposed that a half-logistic time constant (tauL) is a better lusitropic index than tauE at normothermia. In the present study, we investigated whether tauL can remain a superior measure as temperature varies. The isovolumic relaxation LV pressure curves from the minimum of the first time derivative of LV pressure (dP/dtmin) to the LV end-diastolic pressure were analyzed at 30, 33, 36, 38, and 40 degrees C in excised, cross-circulated canine hearts. tauL and tauE were evaluated by curve-fitting using the least squares method and applying the half-logistic equation, P(t) = PA/[1 + exp(t/tauL)] + PB, and the monoexponential equation, P(t) = P0exp(-t/tauE) + Pinfinity. Both tauL and tauE increased significantly with decreasing temperature and decreased with increasing temperature. The half-logistic correlation coefficient (r) values were significantly higher than the monoexponential r values at the 5 above-mentioned temperatures. This implies that the superiority of the goodness of the half logistic fit is not temperature dependent. The half-logistic model characterizes the amplitude and time course of LV pressure fall more reliably than the monoexponential model. Hence, we concluded that tauL is a more useful lusitropic index regardless of temperature. PMID- 18418435 TI - Homocysteine stimulates monocyte chemoattractant protein-1 expression in mesangial cells via NF-kappaB activation. AB - Hyperhomocysteinemia is regarded as an independent risk factor for cardiovascular disorders. Although renal dysfunction or failure is one of the important factors causing hyperhomocysteinemia, the role of homocysteine (Hcy) in the development of glomerulosclerosis is largely unknown. One of the key events in the pathogenesis of glomerulosclerosis is the infiltration of circulating monocytes into affected glomeruli. The objective of the present study was to investigate the effect of Hcy on the expression of monocyte chemoattractant protein-1 (MCP-1) in kidney mesangial cells and the mechanisms involved. Levels of MCP-1 and mRNA were significantly elevated in Hcy-treated rat mesangial cells. This increase was associated with activation of NF-kappaB as a result of increased phosphorylation of the inhibitor protein IkappaBalpha. Monocyte chemotactic activity in these cells was also enhanced. In addition, there was a significant elevation of superoxide anion produced by Hcy-treated cells, which preceded the increased phosphorylation of IkappaBalpha. Addition of superoxide dismutase or NF-kappaB inhibitors to the culture medium abolished Hcy-induced NF-kappaB activation and MCP-1 expression. Taken together, these results indicate that Hcy induced MCP-1 expression in mesangial cells. Such a process was mediated by oxidative stress and NF-kappaB activation. This may further aggravate renal function in patients with hyperhomocysteinemia. PMID- 18418436 TI - Role of endogenous nitric oxide synthase inhibitor in gastric mucosal injury. AB - To explore the role of the endogenous nitric oxide synthase (NOS) inhibitor asymmetric dimethylarginine (ADMA) in gastric mucosal injury, 3 models of gastric mucosal injury induced by ethanol, indomethacin, or cold stress were used in rats. The cultured human gastric mucosal epithelial cell line GES-1 infected by Helicobacter pylori (Hp) was selected to mimic human gastric mucosal injury. Gastric mucosal ulcer index (UI), levels of ADMA and NO, and activity of dimethylarginine dimethylaminohydrolase (DDAH) were determined in the mucosal injury models; in Hp-infected or ADMA-treated GES-1 cells, levels of ADMA, NO, and TNF-alpha and activity of DDAH were measured. The results showed that UI and levels of ADMA were markedly increased and accompanied by significantly decreased DDAH activity in the mucosal injury models. Incubation of GES-1 cells with Hp increased levels of TNF-alpha and ADMA and decreased activity of DDAH. Administration of ADMA also increased levels of TNF-alpha. The results suggest that ADMA plays an important role in facilitating gastric mucosal injury, an effect which is associated with inhibiting NO synthesis and inducing inflammatory reaction. PMID- 18418437 TI - Altered expression of epithelial junctional proteins in atopic asthma: possible role in inflammation. AB - Epithelial cells form a tight barrier against environmental stimuli via tight junctions (TJs) and adherence junctions (AJs). Defects in TJ and AJ proteins may cause changes in epithelial morphology and integrity and potentially lead to faster trafficking of inflammatory cells through the epithelium. Bronchial epithelial fragility has been reported in asthmatic patients, but little is known about the expression of TJ and AJ proteins in asthma. We studied epithelial expression of zonula occludens-1 (ZO-1) and AJ proteins E-cadherin, alpha catenin, and beta-catenin in bronchial biopsies from nonatopic nonasthmatic (healthy) subjects (n = 14), and stable atopic asthmatic subjects (n = 22) at baseline conditions. Immunostaining for these proteins was semi-quantified for separate cellular compartments. E-cadherin, alpha-catenin and beta-catenin were present in the cellular membrane and less in the cytoplasm. Only beta-catenin was present in the nucleus in agreement with its potential function as transcription factor. ZO-1 was present in the apicolateral membrane of superficial cells. alpha Catenin expression was significantly lower in subjects with asthma than without and correlated inversely with numbers of eosinophils within the epithelium. ZO-1 and E-cadherin expression were significantly lower in asthmatic than in nonasthmatic subjects. Expression of beta-catenin was not different. Our results suggest that the lower epithelial alpha-catenin, E-cadherin and (or) ZO-1 expression in patients with atopic asthma contributes to a defective airway epithelial barrier and a higher influx of eosinophils in the epithelium. PMID- 18418438 TI - Effects of amlodipine, diltiazem, and verapamil on the anticonvulsant action of topiramate against maximal electroshock-induced seizures in mice. AB - To assess the effect of 3 calcium channel antagonists (amlodipine, diltiazem, and verapamil) on the anticonvulsant action of topiramate (a new generation antiepileptic drug) in the mouse maximal electroshock seizure (MES) model. Amlodipine (20 mg/kg) significantly enhanced the anticonvulsant activity of topiramate in the MES test in mice, reducing its ED50 value from 54.83 to 33.10 mg/kg (p < 0.05). Similarly, diltiazem (5 and 10 mg/kg) markedly potentiated the antiseizure action of topiramate against MES, lowering its ED50 value from 54.83 to 32.48 mg/kg (p < 0.05) and 28.68 mg/kg (p < 0.01), respectively. In contrast, lower doses of amlodipine (5 and 10 mg/kg) and diltiazem (2.5 mg/kg) and all doses of verapamil (5, 10, and 20 mg/kg) had no significant impact on the antiseizure action of topiramate. Pharmacokinetic verification of the interaction of topiramate with amlodipine and diltiazem revealed that neither amlodipine nor diltiazem affected total brain topiramate concentration in experimental animals, and thus, the observed interactions were concluded to be pharmacodynamic in nature. The favorable combinations of topiramate with amlodipine or diltiazem deserve more attention from a clinical viewpoint because the enhanced antiseizure action of topiramate was not associated with any pharmacokinetic changes in total brain topiramate concentration. PMID- 18418439 TI - Overexpression of Gsalpha compensates for myocyte loss in diabetic cardiomyopathy. AB - The stimulatory G protein Gsalpha transmits signals from activated beta adrenergic receptors via the cyclic AMP-PKA pathway, targeting the key regulatory protein phospholamban. We hypothesized that mice with intrinsic activation of cardiac Gsalpha are resistant to the development of the diabetic cardiomyopathy phenotype. Accordingly, streptozotocin (STZ)-diabetes mellitus was induced in genetically engineered mice with cardiac-specific Gsalpha overexpression and in nontransgenic (NTG) littermates. At 8 weeks, Gsalpha diabetic mice showed no impairment of LV contractility nor increase in myocyte apoptosis, whereas NTG diabetic mice showed a 30% decrease in +dP/dt and -dP/dt with sustained (3-fold) myocyte loss by apoptosis. To assess the level of myocardial reactive oxygen species, we measured malondialdehyde, a surrogate marker of oxidative stress, which was increased in the hearts of NTG and Gsalpha diabetic mice. In addition, chronic hyperglycemia also increased the activity of catalase and superoxide dismutase in the hearts of NTG and Gsalpha diabetic mice. Hearts of NTG diabetic mice, but not Gsalpha mice, showed increased expression of proapoptosis Bax, downregulation in Bcl2, and an increase in the Bax/Bcl2 ratio. Hearts of NTG diabetic mice showed 60% reduction in phosphorylation at the critical Ser16 residue of phospholamban, whereas phosphorylation at Ser16 was restored in hearts of Gsalpha-diabetic mice. We conclude that cardiac-specific overexpression of Gsalpha compensates for the loss of cardiac function in diabetes mellitus. PMID- 18418440 TI - Late protective effect of pharmacological preconditioning with total flavones of rhododendra against myocardial ischemia-reperfusion injury. AB - The aim of the present study was to investigate the protective effect of total flavones of rhododendra (TFR) pharmacological preconditioning against myocardial ischemia-reperfusion (I/R) injury and its probable mechanisms in rats. Rat myocardial I/R injury was induced by ligating and untying the left anterior descending coronary artery. Male Sprague-Dawley rats were anesthetized and the chests were opened. All animals were subjected to 30 min of occlusion and 1 h of reperfusion. Twenty-four hours before the 30-minute occlusion, rats received 3 cycles of 5 min intravenous perfusion of TFR (10, 20, 40 mg/kg) or morphine hydrochloride (0.3 mg/kg) or normal saline interspersed with drug-free periods. Changes in the ST segment of ECG, the content of cardiac troponin I (cTnI), malondialdehyde (MDA), and nitric oxide (NO), and the activity of superoxide dismutase (SOD), lactate dehydrogenase (LDH), creatine phosphokinase (CK), and nitric oxide synthase (NOS) in serum were measured. Infarct size (IS), as a percentage of the area at risk (AAR), was determined by TTC staining. The expression of inducible nitric oxide synthase (iNOS) mRNA in rat myocardium was detected by RT-PCR and the expression of iNOS protein was detected by Western blot. Pretreatment with TFR (10, 20, 40 mg/kg) markedly inhibited I/R-induced ST segment elevation of ECG. TFR (20, 40 mg/kg) pretreatment decreased I/R-induced IS/AAR, markedly inhibited the increase of MDA content and the activity of CK and LDH, and also significantly inhibited the decline of NO content and the activity of NOS and SOD in serum. TFR (40 mg/kg) preconditioning significantly inhibited the increase of serum cTnI induced by I/R injury and increased the expression of iNOS both at mRNA and protein levels in rat myocardium. Our findings indicate that TFR preconditioning has a protective effect against myocardial I/R injury in rats. The cardioprotection involves the stimulation of NO release and the inhibition of lipid peroxidation. PMID- 18418441 TI - Cell-penetrating peptides and proteins: new inhibitors of allergic airways disease. AB - Cell-penetrating peptides (CPPs) or protein transduction domains (PTDs) are peptides that have the ability to efficiently traverse cellular membranes, either alone or in association with molecular cargo. Several naturally occurring PTDs, including those from HIV TAT and Drosophila antennapedia, have this unique activity. Synthetic CPPs, such as polyarginine, also have the ability to enter cells and transport a variety of cargo. While the precise mechanism(s) of cellular entry for individual CPPs may vary, it is likely that uptake is mediated, at least in part, through endocytosis. Moreover, biological activity of cell-penetrating peptides and proteins has been clearly demonstrated in a number of in vitro and in vivo studies. Recently, cell-penetrating proteins targeting the Ras GTPase and the phospholipid kinase PI3K (phosphoinositide 3-kinase) have been shown to inhibit eosinophil trafficking and survival in vitro. These proteins, as well as CPPs targeting the STAT-6 transcription factor or the T-cell costimulatory molecule CTLA-4 (cytotoxic T lymphocyte-associated antigen-4), have also been tested in animal models of asthma. Data from several groups, including ours, indicate that these molecules inhibit airway eosinophilic inflammation, airway hyperresponsiveness (AHR), and mucus production in experimental allergic airways disease. Thus, CPPs targeting these and other signaling molecules may also effectively inhibit allergic airways disease in humans. PMID- 18418442 TI - Understanding exercise behaviour during home-based cardiac rehabilitation: a theory of planned behaviour perspective. AB - Although home-based cardiac rehabilitation (CR) programs have been shown to produce significant increases in exercise capacity, obtaining patient adherence to these programs has been challenging. It is therefore critical to identify key theoretical determinants of exercise during home-based CR in order to inform the development of behavioural interventions that improve adherence. The present study examined the utility of the theory of planned behaviour (TPB) in explaining exercise behaviour during home-based CR. Seventy-six patients who were receiving 6 months of home-based CR completed a TPB questionnaire at the beginning and mid point of the program and a physical activity scale at the mid-point and end of the program. Path analyses showed that attitude and perceived behavioural control significantly predicted intention for both time intervals (baseline to 3 months, and 3 months to 6 months), whereas subjective norm only predicted intention within the 1st 3 months. Intention significantly predicted implementation intention, which, in turn, significantly predicted exercise for both time intervals. Finally, several underlying accessible beliefs were significantly related to exercise for both time intervals. Therefore, results suggest that the TPB is a potentially useful framework for understanding exercise behaviour during home-based CR. PMID- 18418443 TI - Evidence that the etiology of the syndrome containing type 2 diabetes mellitus results from abnormal magnesium metabolism. AB - Evidence is reviewed supporting the presence of an inherited structural defect in the plasma membranes of somatic cells of humans who have type 2 diabetes mellitus and sodium-sensitive essential hypertension. This magnesium-binding defect (MgBD) consists of a decreased content of tightly bound Mg2+ ion in the cell membrane and limits the amount of Mg2+ that enters the cell, some of which combines with ATP4-, produced by the cell, to form MgATP2-, the currency of metabolic energy. Consequently, in both prediabetes and overt diabetes, the intracellular concentration of the interdependent Mg2+ and MgATP2- ions is significantly less than normal. These 2 ions are required as cofactors and (or) substrates for some 300 enzyme systems in human metabolism, many of which are involved with insulin. Thus the decreased activities of particular ones of these enzyme systems due to the decreased intracellular [Mg2+] and its dependent [MgATP2-] are responsible for (i) insulin resistance and (ii) decreased insulin secretion and (or) production, the 2 pathophysiological processes required for the occurrence of type 2 diabetes mellitus. These 2 processes can account for all of the morbid symptoms associated with this disease. Thus, the decreased intracellular concentration of the interdependent Mg2+ and MgATP2- ions constitutes the etiology of genetic predisposition to type 2 diabetes mellitus and can be corrected by 2 identified peptide Mg2+-binding promoters that are derived from the carboxyl terminal of the tachykinin substance P and occur in normal blood plasma. Decreased intracellular [Mg2+] and [MgATP2-] can also result from a dietary deficiency of magnesium or from an abnormal accumulation of saturated fatty acids in cell membranes, which inhibits the entrance of Mg2+ into the cell; thus it is also the etiology not only of diabetes caused by magnesium deficiency, but also of the "lipotoxic" type 2 diabetes mellitus. Although these pathologies cannot be corrected by the Mg2+-binding promoters, they can be corrected, respectively, by dietary magnesium supplementation or by exercise plus dietary caloric and lipid restriction. Theoretically, the disease syndrome containing type 2 diabetes mellitus may involve approximately 30% of the population. PMID- 18418444 TI - Effects of sex hormone levels on aortic vascular reactivity and variables associated with the metabolic syndrome in sucrose-fed female rats. AB - We studied the effect of varying levels of sex hormones, induced by ovariectomy and administration of testosterone or estradiol, on aortic reactivity in female rats with metabolic syndrome (MS) induced by a sucrose diet. Vasoreactivity of aortic rings, blood pressure, intra-abdominal fat, serum triglycerides, nitrates and nitrites, and TBARS were evaluated. Intact MS and ovariectomized MS had higher BP than intact control (C) and ovariectomized C, respectively; estradiol administration decreased BP in ovariectomized MS but not in ovariectomized C. Triglycerides and fat were both higher in MS. Triglycerides were not modified by surgery or hormone treatment, but ovariectomy increased fat. When ovariectomy was combined with hormones, however, fat was reduced to the level of intact rats. Ovariectomy decreased, but hormones increased, serum nitrates and nitrites. Vasoconstriction was larger in intact MS and ovariectomized MS + testosterone aortas than in intact C and ovariectomized C + testosterone, respectively. Vasodilation was reduced in intact MS and ovariectomized MS + testosterone compared with intact C, ovariectomized C + testosterone, ovariectomized MS, and ovariectomized MS + estradiol. The results suggest endothelial dysfunction in intact MS and ovariectomized MS + testosterone, but protection by ovariectomy + estradiol in MS due to hormones. Indomethacin reduced all contractions, but the effect was greater in estradiol-treated rats. L-NAME increased contractility, more in the ovariectomized C and MS groups and less in the estradiol-treated groups. PMID- 18418445 TI - Na-P(i) cotransporter type I activity causes a transient intracellular alkalinization during ATP depletion in rabbit medullary thick ascending limb cells. AB - The cellular pathophysiology of renal ischemia-reperfusion injury was investigated in primary cell cultures from rabbit medullary thick ascending limb (MTAL). Metabolic inhibition (MI) was achieved with cyanide and 2-deoxyglucose. Sixty minutes of MI caused a profound but reversible decrease in intracellular concentration of ATP ([ATP]i). Intracellular pH (pHi) first decreased after initiation of MI, followed by a transient alkalinization. When [ATP]i reached its lowest value (<1% of control), the cells slowly acidified to reach a stable pHi of 6.92 after 50 min of MI. In the presence of EIPA (10 micromol/L), the pattern of changes in pHi was unchanged and acidification was not increased, indicating that the Na+/H+ exchangers were inactive during ATP depletion. When inorganic phosphate (P(i)) or Na+ was omitted from the apical solutions during MI, the transient alkalinization was no longer observed and the cytosol slowly acidified. Experiments on Na+-dependent alkalinizations revealed the presence of a Na-P(i) cotransporter in the apical cell membrane. With indirect immunofluorescence, the Na-P(i) cotransporter expressed in these primary cell cultures could be identified as Na-P(i) type I. Although the exact physiological role of Na-P(i) type I still is unresolved, these experiments demonstrate that apical Na-P(i) type I activity is increased at the onset of ATP depletion in MTAL cells. PMID- 18418446 TI - Adenosine reduces the reverse mode of the Na+/Ca(2+) exchanger in ferret cardiac fibres. AB - The aim of this study was to investigate the effects of adenosine on reverse mode Na+/Ca(2+) exchange. In intact ferret cardiac trabeculae, Na+-free contractures were investigated after treating preparations with ryanodine, a sarcoplasmic reticulum Ca(2+) -channel inhibitor, and thapsigargin, a sarcoplasmic reticulum Ca(2+) -pump inhibitor added to suppress the sarcoplasmic reticulum function. The effects of adenosine (50-100 nmol/L), adenosine deaminase (ADA, 0.1-0.5 U/L), the A1 and A2A receptor agonists CCPA (3-100 nmol/L) and CGS 21680 (25-100 nmol/L), and the A1 and A2A receptor antagonists DPCPX (25 nmol/L) and ZM 241385 (25 nmol/L) were tested on Na+-free contractures. The application of adenosine (50 100 nmol/L) had no significant effect on the characteristics of the Na+-free contractures. However, the results show that treatment with ADA (0.3 U/L), adenosine (> or =50 nmol/L) and CCPA, a specific A1 receptor agonist (3-100 nmol/L), all reduced the Na+-free contracture amplitude. In the presence of ADA, the effects of adenosine and CCPA were also reduced by a specific antagonist of A1 receptors (DPCPX, 25 nmol/L). Furthermore, adenosine, ADA, and CCPA did not affect the properties of the contractile apparatus in Triton-skinned fibres. It is therefore proposed that endogenous adenosine reduced the reverse mode of the Na+/Ca(2+) exchanger by acting on A1 receptors present in the sarcolemmal membrane. PMID- 18418448 TI - Alteration of superoxide- and nitric oxide-mediated antimicrobial function of macrophages by in vivo cocaine exposure. AB - Cocaine is a popular drug of abuse and despite impressive advances in the understanding of its physiological, pharmacological, and toxic effects, its mechanism of immunosuppression at the cellular level is not well understood. In this paper we report the role of effector molecules like superoxide and nitric oxide in the antibacterial function of macrophages exposed to acute and chronic doses of cocaine in vivo. Bacterial killing by acute cocaine-exposed macrophages (ACE-Mphis) increased significantly, with a concomitant rise in respiratory burst and generation of superoxide and nitric oxide, compared with control macrophages. In contrast, chronic cocaine-exposed macrophages (CCE-Mphis) exhibited limited antimicrobial activity, which correlated closely with diminished respiratory burst and reduced production of superoxide and nitric oxide. Further, a killing assay was carried out in the presence of N(G)-methyl-L-arginine acetate, an inhibitor of iNOS, to evaluate the role of nitric oxide in the killing process. The results obtained indicate that while about 30% killing of input bacteria by control and ACE-Mphis was attributable to NO-mediated killing, only about 6% killing from NO was found with CCE-Mphis. The findings indicate that acute exposure to cocaine possibly caused upregulation of enzymes responsible for the generation of ROI (reactive oxygen intermediates) and RNI (reactive nitrogen intermediates), leading to enhanced antimicrobial function. On the other hand, chronic exposure to cocaine impaired the oxygen-dependent microbicidal capacity of macrophages, possibly through impaired expression of enzymes responsible for ROI and RNI formation. Proinflammatory cytokines may play a key role in cocaine mediated immunosuppression, since exposure of macrophages to cocaine impairs the ability of the cells to produce these cytokines. PMID- 18418447 TI - Renal effect of meloxicam versus ketoprofen in anaesthetized pseudo-normovolaemic piglets. AB - Due to renal COX-2 constitutive expression, meloxicam is presumably deleterious for kidney function in critical situations. The present study investigates the influence of intravenous meloxicam on renal parameters and compares it with a nonselective COX inhibitor, ketoprofen. Piglets (n = 6 in each group) were treated with ketoprofen (2 mg.kg(-1)), meloxicam (0.2 mg.kg(-1)), or saline at the beginning of anaesthesia. Under intravenous anaesthesia, pigs were instrumented for cardiovascular, respiratory, and renal function evaluation, including urinary flow (UF), glomerular filtration rate (GFR), and renal blood flow (RBF). After baseline data collection (U0), data collection consisted of six 20-minute periods (U1 to U6). In all groups, the time course of cardiovascular and respiratory parameters remained within normal ranges. A small decrease in cardiac output and an increase in mean systemic arterial blood pressure (p = 0.002) occurred in all groups. In the placebo group, a similar decrease was observed for RBF and cardiac output, with troughs of -10.1% +/- 6.8%, and -12.9% +/- 3.2%, respectively. GFR and UF, however, remained stable over time in this group. Ketoprofen significantly decreased UF (-29.3% +/- 5.5% max at U3), with similar decreases in GFR and RBF. Meloxicam induced a transient (at U2) and small decrease in UF with no difference, at any time point, with the placebo group. The renal effects of meloxicam appear minimal and transient in anaesthetized piglets. This study demonstrates the safety of meloxicam for preemptive surgical analgesia under conditions of normovolaemia. Fluid therapy appears recommended to prevent any renal dysfunction. PMID- 18418449 TI - [Primary adenoid cystic carcinoma of esophagus]. AB - We presented the case of a man of 86 years with 6 months of progressive dysphagia, weight loss and edema of lower limbs. He had both severe microcitic anemia and hypoalbuminemia. The contrasted x-ray showed the esophagus with irregular edges that compromised their cervical and distal portions. In the computerized axial tomography cervical adenopathies, bilateral pleural effusion and pronouncing esophagic thickening were evident. In endoscopy dispersed elevated lesions were observed from the underlying area of the cricopharinx, some with nodular and tumor like aspect, that distributed throughout the esophagus until an area of stenosis (32 cm of dental arches); the estenosis was tapestried with an irregular and easy bleeding mucosa. The histological study revealed adenoid cystic carcinoma of esophagus, with positive inmunocytochemical to cytokeratin. We showed the clinical-pathological findings and images of this case and we reviewed reports of this rare entity. PMID- 18418450 TI - [Prebiotic inulin/oligofructose in Yacon root (Smallanthus sonchifolius), phytochemistry and standardization as basis for clinical and pre-clinical research]. AB - INTRODUCTION: Experiments have demonstrated that the association between inulin and oligofructose can prevent colitis by modifying the intestinal microflora by acting as a prebiotic. It has also been found that this association is naturally present in the roots of the Yacon root (Smallanthus sonchifolius) making phytochemical and standardization studies well worth pursuing. These studies could contribute to the safe use of improved strains of Yacon, to significant savings in health resources, and to promoting additional clinical and pre clinical research. So far, studies have been made on organically cultivated Yacon in the coast, on varieties adapted from those cultivated in the Andes. The aim is to promote consumption of Yacon in the sectors of society that need it most. OBJECTIVE: To determine the general chemical composition of the Yacon tuberous root cultivated in the coast by means of a phytochemical screening, as well as standardizing its inulin content in order to contribute knowledge on its conditions of use. METHODS: The roots harvested in the Lima Botanical Garden of the National Health Institute were used to prepare aqueous extracts through decoction and juice for the phytochemical screening with conventional reagents. The aqueous extracts were standardized for insulin content by using a fructose standard method and a validated analytical technique. The samples were read in a TU-1810S Split Beam UV-VIS spectrophotometer. RESULTS: The trials confirmed the presence of phenolic compounds, flavonoids, alkaloids, steroids, glycosides and carbohydrates. The standardization found values of 7,8% inulin in the hydrolyzed extract and of 7, 01% in the non-hydrolyzed extract. CONCLUSIONS: The extracts of the Yacon roots were screened and standardized Ander known cultivation conditions. The possibilities of its use are being considered due to its prebiotic properties. These properties are considered useful in treating certain types of colitis by stimulating the growth of the Lactobacillus and Bifidobacterium microflora. Saving of health resources would be possible and further clinical and pre-clinical research would be encouraged. PMID- 18418451 TI - [Most frequent mistakes in medical statistics]. PMID- 18418452 TI - [2008, Year of The XXI National Congress of Gastroenterology]. PMID- 18418453 TI - [Mirizzi syndrome associated with complicated cholelitiasis in the elderly patient. Diagnosis and laparoscopic treatment]. AB - INTRODUCTION: The Mirizzi Syndrome (MS) is a rare variation of cholelitiasis, in which a calculus impacted in the Hartmann Pouch compresses the biliary pathway triggering an obstructive jaundice, frequently followed by inflammatory phenomenon and a number of complications. OBJECTIVE: To establish the frequency of the Mirizzi Syndrome in complicated cholelitiasis in the elderly patient and analyze the most suitable diagnosis and treatment options. PATIENTS AND METHOD: Cases of Mirizzi Syndromes in symptomatic inflammatory biliary lithiasis were selected in a five-year period among patients older than 70, who had undergone urgent operation. Epidemiology, private hospital, diagnosis, operating variables, morbility and hospitalization were analyzed. RESULTS: Twelve (12) patients with Mirizzi Syndrome were detected, with an average age of 77.2 in 197 cases of complicated cholelitiasis. A laparoscopic examination was made in 67% of the cases, with a conversion rate of 50%. The most frequent finding was the biliary extrinsic compression without Csendes-I fistula (58%). The postsurgical morbility was of 58%, including two deaths (17%) and the medium-term postoperative hospital stay was of 9.5 days. CONCLUSIONS: This research emphasizes the importance of the Mirizzi Syndrome (MS) in the symptomatic cholelitiasis in geriatric patients. This syndrome usually acts like a biliar surgical emergency and the early diagnosis of an elderly patient with jaundice is the key that enables immediate biliary decompression through a cholecystectomy. Management choices are in debate; however, due to its advantages in cases of elderly patients, a laparoscopic treatment is proposed, unless strictly contraindicated. PMID- 18418454 TI - [Evaluation of the effect of nitazoxanide compared with placebo in patients with flatulence at the ambulatory consultation of the Central Clinic of Inppares Lima]. AB - Flatulence is a very common complaint related to functional gastrointestinal disorders. We know functional disorders is the main cause of consultation in gastroenterology offices. We don't know the exact reason of flatulence, but the intestinal fermentative microbiota could be an important etiologic factor. The objective of the study is to evaluate the effectiveness of the Nitazoxanida, on the clinical improvement of the flatulence in a group of patients of ambulatory consultation with respect to another group that receives placebo. The present article, is a controlled randomized clinical study designed to double blind, in whom 120 patients with flatulence criteria participate, of which 60 patients received Nitazoxanida 500mg, every 12 hours by 3 days, and the next 60 patients received placebo every 12 hours by 3 days, after one week were reevaluated, and they were put under a test of perception about clinical improvement (Jerome Frank). The Nitazoxanida group and the placebo group were very similar in age, sex, symptoms to the entrance, presence of anxiety, depression and upheavals of the dream. In the Nitazoxanida group was an average of improvement of 4.02 (75.31%) DS 0.94 and with placebo 2.35 (19.58%) D.S. 0.63. with percentage of 0.001 error. Being the perception of global improvement in the Nitazoxanida group 91.67% and in the placebo group 36.67%. Which is statistically significant. The study conclude that Nitozoxanide group produce a significative improvement in the perception of relief of flatulence in comparison to the placebo group. The study sets out a new therapeutic indication of the active principle Nitazoxanide, in flatulence. We found is a high prevalence of anxiety, depression and of upheavals of the dream in patients with flatulence. PMID- 18418455 TI - [Fibroepithelial lesions of the anal canal: therapeutic technique with rubber band ligation]. AB - When fibroepithelial lesions of the anal canal increase in size and bleed, they can produce tenesmus and a sensation of a foreign body in the anus. Generally, these lesions are surgically excised. We present an original non-surgical technique using rubber band ligation. In the Gastroenterology Department of the "Edgardo Rebagliati Martins" National Hospital (EsSALUD) between June 2000, and June, 2007, 16 patients with anal fibroepithelioma were included: 6 men and 10 women, with an average age of 44, aged between 21 to 75, and 44% aged 20 to 29. All of these patients (100%) had anal tumors, 50% had prolapses, 28% had anal pain and, 12% intermittent slight bleeding. The lesions had a polypoid appearance and sizes ranged from 1-5 cm. The technique consists in injecting 2% Xylocaine to the lesion, which is then retracted with surgical tweezers through the ring of the device holding the rubber band, which is then released leaving a safety margin of 5 to 10 mm over the skin. Biopsies of 11 lesions were taken and their analysis confirmed that there were 10 fibroepitheliomas and squamous epithelium. There was involution of the lesions due to ischemic necrosis in subsequent days. Patients only required oral analgesics for the first hours after procedure; only one patient presented an acute anal fissure. This non-surgical alternative performed on outpatient basis has proved to be effective and exempt from further complications. PMID- 18418456 TI - [Practices, knowledge and attitudes about human hydatidosis in Peru]. AB - Between July 2005 and June 2006, a case-control study was carried out to identify risk factors for hydatidosis in Lima, Peru. As a result, 32 cases were matched according to age, sex and birthplace in 64 controls. The participants were interviewed using a questionnaire designed to evaluate environmental and behavior factors associated to hydatidosis. By using Multiple Conditional Logistic Regression, it was determined that the owners of >or=10 dogs in a rural area (aOR=8.7; 95% Cl=1.3-57.5) and those raising sheep (aOR=5.9; 95% Cl=1.2-28.1) were independently related to a higher risk of hydatidosis. The belief that food may transmit hydatidosis (aOR=0.1; 95% Cl=0.01-0.7) and the activity of raising goats (aOR=0.02; 95% Cl=0.001-0.6) were inversely associated to hydatidosis. Preventive measures to reduce the transmission of hydatidosis to human beings in endemic areas in Peru require: limiting the number of dogs kept in each household, regularly controlling parasites and preventing access of dogs to human food and drinking water; all of this in addition to educational campaigns to change the practices that lead to the transmission of hydatidosis. PMID- 18418457 TI - [Pancreatic lipoma: presentation of a case and review of literature]. AB - Diagnosis of pancreatic lipoma, is very rare. We presented the case of a 51 years woman who show a tumor of 9x8 cm. at the head and neck of the pancreas. A pancreatoduodenectomy was performed with favorable result. We discuss the therapeutic decision. PMID- 18418458 TI - [Pancreatic pseudoaneurysm: surgical treatment]. AB - The pseudoaneurysm is a rare complication of severe acute Pancreatitis and is more prevalent in alcoholic patients. This case report describes a 44-year-old male patient with a history of chronic alcoholism with pancreatic pseudoaneurysm by arterial erosion, which was diagnosed by CT imaging and US Doppler abdominal and successfully treated with transfixiant ligation of splenic artery with additional surgical drainage by Foley catheter. PMID- 18418459 TI - [Intrahepatic lithiasis]. AB - Presence of stones in the intrahepatic bile ducts is a common problem especially in East Asia. We report two cases: the first is a man who through several years had repeated episodes of obstructive jaundice. He underwent multiple procedures due to the recurrences. The second case is a woman who had recurrences and serious complications. A detailed evaluation and early diagnosis of each case are essential for a good treatment, which is challenging due to the high rate of recurrence and complications. PMID- 18418461 TI - [Lipecs and Scielo Peru: sources of information on health research in Peru]. PMID- 18418460 TI - [Tumoral microembolism and cor-pulmonar as manifestation of hepatocelular carcinoma]. AB - The tumour pulmonary micro-embolism is a rare condition characterized by the occlusion of pulmonary small arteries, arteriolas, and alveolar capillaries septales, accompanied of trombosis. Occasionally the development of pulmonary hypertension is the first manifestation of an occult neoplasia, in series of autopsies, an incidence from 3 to 26% has been reported in solid tumors, being clinical evident in 8%. Few cases have documented the development of this condition in patients with carcinoma hepatocelular, we report the case of a 16 year-old male who comes to the emergency with signs of cardiac insufficiency and cor pulmonare whose anatomopatological study confirmed a tumour massive microembolic compromise at pulmonary level and hepatocarcinoma. PMID- 18418462 TI - Mentoring health information professionals in the Department of Veterans Affairs. AB - As a major employer of health information professionals, the VA faces significant recruitment and retention challenges. The authors evaluated mentoring as a retention tool through a review of existing literature and the retrospective review of a VA health information management mentoring program. The literature review showed a link between employer mentorship and employee retention, regardless of the nature and structure of the mentoring relationship. Most organizations support employees who are willing to serve as mentors through increased compensation, recognition, and other types of support. No literature was found that studied retention rates for more than three years after a mentoring experience. The review of the VA mentoring program showed increased retention in the three years following enrollment in the program, but the increase was not statistically significant. The review did not demonstrate improvement in retention over a seven-year period. The combined evaluation gives mixed findings for mentorship as a retention tool and demonstrates the need for more research on the topic. PMID- 18418464 TI - Avian influenza, domestic ducks and rice agriculture in Thailand. AB - Highly pathogenic avian influenza (HPAI) caused by H5N1 viruses has become a global scale problem which first emerged in southern China and from there spread to other countries in Southeast and East Asia, where it was first confirmed in end 2003. In previous work, geospatial analyses demonstrated that free grazing ducks played critical role in the epidemiology of the disease in Thailand in the winter 2004/2005, both in terms of HPAI emergence and spread. This study explored the geographic association between free grazing duck census counts and current statistics on the spatial distribution of rice crops in Thailand, in particular the crop calendar of rice production. The analysis was carried out using both district level rice statistics and rice distribution data predicted with the aid of remote sensing, using a rice-detection algorithm. The results indicated a strong association between the number of free grazing ducks and the number of months during which second-crop rice harvest takes place, as well as with the rice crop intensity as predicted by remote sensing. These results confirmed that free grazing duck husbandry was strongly driven by agricultural land use and rice crop intensity, and that this later variable can be readily predicted using remote sensing. Analysis of rice cropping patterns may provide an indication of the location of populations of free grazing ducks in other countries with similar mixed duck and rice production systems and less detailed duck census data. Apart from free ranging ducks and rice cropping, the role of hydrology and seasonality of wetlands and water bodies in the HPAI risk analysis is also discussed in relation to the presumed dry season aggregation of wild waterfowl and aquatic poultry offering much scope for virus transmission. PMID- 18418463 TI - p16INK4a gene promoter hypermethylation in mucosa as a prognostic factor for patients with colorectal cancer. AB - Low gene expression of folylpolyglutamate synthase (FPGS) in colorectal mucosa correlates with low folate levels and poor survival of colorectal cancer (CRC) patients. Because gene-specific hypermethylation is affected by the folate level, the hypermethylation status in mucosa may also be linked to clinical outcome of CRC patients. The tumor suppressor gene p16INK4a (p16) regulates the cell cycle and angiogenic switch. In human neoplastic tissues, the main mechanism of p16 inactivation is promoter methylation. The aim of the study was to determine whether hypermethylation of the p16 promoter could be detected in mucosa of CRC patients (n = 181) and to analyze if hypermethylation was related to survival. The relation between p16 hypermethylation and expression of FPGS and two other folate-associated genes, reduced folate carrier 1 (RFC-1), and thymidylate synthase (TS), was analyzed (n = 63). The results showed that p16 was hypermethylated in 65 (36%) of the mucosa samples and that hypermethylation was age-related (P = 0.029). After adjustment for known risk factors, Cox regression analysis showed that Dukes' A-C patients with p16 hypermethylation in mucosa had an increased risk of cancer-related death (hazard ratio = 2.9, P = 0.007) and shorter disease-free survival (hazard ratio = 2.5, P = 0.015) compared with patients with no p16 hypermethylation. RFC-1 and FPGS gene expression levels were significantly correlated in patients lacking p16 hypermethylation in mucosa (P = 0.0003), but not at all correlated in patients having hypermethylation in mucosa (P = 1.0). In conclusion, p16 hypermethylation in mucosa of CRC patients was identified as an independent prognostic parameter for cancer-specific survival as well as an independent predictor of DFS. The results suggest that there might be a connection between folate-associated gene expression and p16 methylation status. PMID- 18418465 TI - Population-Calibrated Gene Characterization: Estimating Age at Onset Distributions Associated With Cancer Genes. AB - Phenotypic characterization of rare disease genes poses a significant statistical challenge, but the need to do so is clear. Clinical management of patients carrying a disease gene depends crucially on an accurate characterization of the genetically predisposed disease, including its likelihood of occurrence among mutation carriers, natural history, and response to treatment. We propose a formal yet practical method for controlling for bias due to ignoring ascertainment, defined as the sampling mechanism, when quantifying the association between genotype and disease using data on high-risk families. The approach is more statistically efficient than conditioning on the variables used in sampling. In it, the likelihood is adjusted by a factor that is a function of sampling weights in strata defined by those variables. It requires that these variables and the sampling probabilities in the strata they define either are known or can be estimated. The latter requires a second, population-based dataset. As an example, we derive ascertainment-corrected estimates of penetrance for the breast cancer susceptibility genes BRCA1 and BRCA2. The Bayesian analysis that we use incorporates a modified segregation model and prior data on penetrance derived from the literature. Markov chain Monte Carlo methods are used for inference. PMID- 18418467 TI - Analytical reconstruction of 4D dynamic cardiac SPECT with noise-reduction and non-uniform attenuation compensation. AB - BACKGROUND: This study focuses on quantitative reconstruction of dynamic cardiac SPECT. MATERIAL AND METHODS: The Karhunen-Loeve (KL) transform is first applied to gated frames or sinogram data to de-correlate the dynamic information along the time direction. Then the Poisson noise is treated by a penalized weighted least-squares minimization, followed by an analytical inversion of the treated sinograms with attenuation compensation via the Novikov formula. The reconstruction is completed by inverse KL transform. RESULTS: Dynamic or gated cardiac sinograms were simulated from the NCAT phantom mimicking the human torso, and their reconstructions by the presented method showed significant improvement over the conventional methods of frame-by-frame reconstruction. CONCLUSION: This analytical reconstruction of dynamic SPECT is consistent and efficient with a good potential for practical use. PMID- 18418466 TI - Classification of Missense Mutations of Disease Genes. AB - Clinical management of individuals found to harbor a mutation at a known disease susceptibility gene depends on accurate assessment of mutation-specific disease risk. For missense mutations (MMs)-mutations that lead to a single amino acid change in the protein coded by the gene-this poses a particularly challenging problem. Because it is not possible to predict the structural and functional changes to the protein product for a given amino acid substitution, and because functional assays are often not available, disease association must be inferred from data on individuals with the mutation. Inference is complicated by small sample sizes and by sampling mechanisms that bias toward individuals at high familial risk of disease. We propose a Bayesian hierarchical model to classify the disease association of MMs given pedigree data collected in the high-risk setting. The model's structure allows simultaneous characterization of multiple MMs. It uses a group of pedigrees identified through probands tested positive for known disease associated mutations and a group of test-negative pedigrees, both obtained from the same clinic, to calibrate classification and control for potential ascertainment bias. We apply this model to study MMs of breast-ovarian susceptibility genes BRCA1 and BRCA2, using data collected at the Duke University Medical Center in Durham, North Carolina. PMID- 18418468 TI - PRODUCTIVITY SPILLOVERS IN HEALTHCARE: EVIDENCE FROM THE TREATMENT OF HEART ATTACKS. AB - A large literature in medicine documents variation across areas in the use of surgical treatments that is unrelated to outcomes. Observers of this phenomena have invoked "flat of the curve medicine" to explain these facts, and have advocated for reductions in spending in high-use areas. In contrast, we develop a simple Roy model of patient treatment choice with productivity spillovers that can generate the empirical facts. Our model predicts that high-use areas will have higher returns to surgery, better outcomes among patients most appropriate for surgery, and worse outcomes among patients least appropriate for surgery, while displaying no relationship between treatment intensity and overall outcomes. Using data on treatments for heart attacks, we find strong empirical support for these and other predictions of our model, and reject alternative explanations such as waste or supplier induced demand, for geographic variation in medical care. PMID- 18418470 TI - The Markers and Meanings of Growing Up: Contemporary Young Women's Transition from Adolescence to Adulthood. AB - Growing up in the shadow of the women's movement has created contradictory life course and identity possibilities for young women. Although prior research has examined the formal markers of adulthood, we know little about how young women themselves perceive these markers. Forty-two in-depth interviews revealed that the subjective meanings of young women's transition to adulthood are actually far more complex than previously assumed. While becoming a parent and becoming financially independent were seen by interviewees as reflecting an adult orientation, completing schooling was tied to class-differentiated views of growing up. In addition, beginning full-time work was subjectively linked to future career uncertainty, and getting married did not diminish young women's emphasis on self-development and independence from men. Taken together, these findings indicate that there is a disjuncture between women's objective and subjective transition to adulthood. This study suggests that our previous understandings of the transition to adulthood do not reflect the full complexity of how young women subjectively experience it or the extent to which class impacts these perceptions. PMID- 18418473 TI - Exploring cross-linguistic vocabulary effects on brain structures using voxel based morphometry. AB - Given that there are neural markers for the acquisition of a non-verbal skill, we review evidence of neural markers for the acquisition of vocabulary. Acquiring vocabulary is critical to learning one's native language and to learning other languages. Acquisition requires the ability to link an object concept (meaning) to sound. Is there a region sensitive to vocabulary knowledge? For monolingual English speakers, increased vocabulary knowledge correlates with increased grey matter density in a region of the parietal cortex that is well-located to mediate an association between meaning and sound (the posterior supramarginal gyrus). Further this region also shows sensitivity to acquiring a second language. Relative to monolingual English speakers, Italian-English bilinguals show increased grey matter density in the same region.Differences as well as commonalities might exist in the neural markers for vocabulary where lexical distinctions are also signalled by tone. Relative to monolingual English, Chinese multilingual speakers, like European multilinguals, show increased grey matter density in the parietal region observed previously. However, irrespective of ethnicity, Chinese speakers (both Asian and European) also show highly significant increased grey matter density in two right hemisphere regions (the superior temporal gyrus and the inferior frontal gyrus). They also show increased grey matter density in two left hemisphere regions (middle temporal and superior temporal gyrus). Such increases may reflect additional resources required to process tonal distinctions for lexical purposes or to store tonal differences in order to distinguish lexical items. We conclude with a discussion of future lines of enquiry. PMID- 18418472 TI - Perigenual anterior cingulate morphology covaries with perceived social standing. AB - Low socioeconomic status (SES) increases the risk for developing psychiatric and chronic medical disorders. A stress-related pathway by which low SES may affect mental and physical health is through the perception of holding a low social standing, termed low subjective social status. This proposal implicates overlapping brain regions mediating stress reactivity and socioemotional behaviors as neuroanatomical substrates that could plausibly link subjective social status to health-related outcomes. In a test of this proposal, we used a computational structural neuroimaging method (voxel-based morphometry) in a healthy community sample to examine the relationships between reports of subjective social status and regional gray matter volume. Results showed that after accounting for potential demographic confounds, subclinical depressive symptoms, dispositional forms of negative emotionality and conventional indicators of SES, self-reports of low subjective social status uniquely covaried with reduced gray matter volume in the perigenual area of the anterior cingulate cortex (pACC)-a brain region involved in experiencing emotions and regulating behavioral and physiological reactivity to psychosocial stress. The pACC may represent a neuroanatomical substrate by which perceived social standing relates to mental and physical health. PMID- 18418474 TI - Effectiveness of frequency shifted feedback at reducing disfluency for linguistically easy, and difficult, sections of speech (original audio recordings included). AB - Frequency shifted feedback (FSF) induces fluency when presented to speakers who stutter. This study examined whether FSF was more effective at removing disfluencies on easy or on difficult stretches of speech (where difficulty was defined with respect to utterance and word length). There were more disfluencies on the difficult stretches than on the easy stretches. There were significantly fewer disfluencies under FSF than in normal listening conditions (indicating that FSF improved fluency). There was no interaction between difficulty of material and type of feedback when disfluency rate was used as the dependent variable, suggesting that targeting FSF on easy stretches of speech is as effective as targeting it on difficult stretches. The original audio data are provided in this report and can be used by readers to check for themselves the characteristics of voice control that alter when FSF is delivered. PMID- 18418469 TI - Retinoid-related Orphan Receptors (RORs): Roles in Cellular Differentiation and Development. AB - Retinoid-related orphan receptors RORalpha, -beta, and -gamma are transcription factors belonging to the steroid hormone receptor superfamily. During embryonic development RORs are expressed in a spatial and temporal manner and are critical in the regulation of cellular differentiation and the development of several tissues. RORalpha plays a key role in the development of the cerebellum particularly in the regulation of the maturation and survival of Purkinje cells. In RORalpha-deficient mice, the reduced production of sonic hedgehog by these cells appears to be the major cause of the decreased proliferation of granule cell precursors and the observed cerebellar atrophy. RORalpha has been implicated in the regulation of a number of other physiological processes, including bone formation. RORbeta expression is largely restricted to several regions of the brain, the retina, and pineal gland. Mice deficient in RORbeta develop retinal degeneration that results in blindness. RORgamma is essential for lymph node organogenesis. In the intestine RORgamma is required for the formation of several other lymphoid tissues: Peyer's patches, cryptopatches, and isolated lymphoid follicles. RORgamma plays a key role in the generation of lymphoid tissue inducer (LTi) cells that are essential for the development of these lymphoid tissues. In addition, RORgamma is a critical regulator of thymopoiesis. It controls the differentiation of immature single-positive thymocytes into double-positive thymocytes and promotes the survival of double-positive thymocytes by inducing the expression of the anti-apoptotic gene Bcl-X(L). Interestingly, all three ROR receptors appear to play a role in the control of circadian rhythms. RORalpha positively regulates the expression of Bmal1, a transcription factor that is critical in the control of the circadian clock. This review intends to provide an overview of the current status of the functions RORs have in these biological processes. PMID- 18418475 TI - Facilities to assist people to research into stammered speech. AB - The purpose of this article is to indicate how access can be obtained, through Stammering Research, to audio recordings and transcriptions of spontaneous speech data from speakers who stammer. Selections of the first author's data are available in several formats. We describe where to obtain free software for manipulation and analysis of the data in their respective formats. Papers reporting analyses of these data are invited as submissions to this section of Stammering Research. It is intended that subsequent analyses that employ these data will be published in Stammering Research on an on-going basis. Plans are outlined to provide similar data from young speakers (ones developing fluently and ones who stammer), follow-up data from speakers who stammer, data from speakers who stammer who do not speak English and from speakers who have other speech disorders, for comparison, all through the pages of Stammering Research. The invitation is extended to those promulgating evidence-based practice approaches (see the Journal of Fluency Disorders, volume 28, number 4 which is a special issue devoted to this topic) and anyone with other interesting data related to stammering to prepare them in a form that can be made accessible to others via Stammering Research. PMID- 18418476 TI - Organ transplantation in HIV-infected individuals: The time has come - should there be criteria? PMID- 18418477 TI - Ode to oseltamivir and amantadine? PMID- 18418478 TI - Ethical and scientific issues surrounding solid organ transplantation in HIV positive patients: Absence of evidence is not evidence of absence. AB - End-stage liver disease is emerging as a leading cause of death among HIV positive patients. Historically, an HIV diagnosis was a contraindication for a liver transplant; however, because of the efficacy of highly active antiretroviral therapy (HAART), HIV-positive patients have one-year, two-year, and three-year post-transplantation survival rates similar to that of HIV negative patients. Based on this evidence, HIV-positive patients are now considered eligible for transplantation. However, newly emerging guidelines include the stipulation that HIV-positive patients must be on HAART to be placed on a waiting list for transplantation. The purpose of the present paper is to evaluate the scientific and ethical probity of requiring HIV-positive patients to be on HAART as a condition for being on a liver transplant waiting list. It is argued that the emphasis should be placed on the probability of post transplantation HAART tolerance, and that concerns about pretransplantation HAART tolerance are of secondary importance. PMID- 18418479 TI - Pneumococcal conjugate vaccination in Canadian infants and children younger than five years of age: Recommendations and expected benefits. AB - INTRODUCTION: Streptococcus pneumoniae infection may result in invasive pneumococcal disease (IPD), such as bacteremia, meningitis and bacteremic pneumonia, or in non-IPD, such as pneumonia, sinusitis and otitis media. In June 2001, a heptavalent pneumococcal conjugate vaccine (PCV7) (Prevnar, Wyeth Pharmaceuticals, Canada) was approved for use in children in Canada. The objective of the present paper is to review S pneumoniae-induced disease incidence and vaccine recommendations in Canadian infants and children younger than five years of age. Particular attention is given to the expected benefits of vaccination in Canada based on postmarketing data and economic modelling. METHODS: Searches were performed on PubMed and Web of Science databases and specific Canadian journals using the key words 'pneumococc*', 'vaccine', 'conjugate', 'infant' and 'Canadian'. RESULTS AND DISCUSSION: PCV7 appears to be safe and effective against IPD and non-IPD in children younger than five years of age and, more importantly, in children younger than two years of age (who are at highest risk for IPD). An examination of postmarketing data showed a reduction in incidence of pneumococcal disease in age groups that were vaccinated and in older age groups, indicating the likelihood of herd protection. Concurrently, there was a reduction in the occurrence of antimicrobial-resistant isolates. CONCLUSIONS: The results from the present review suggest that PCV7 is currently benefiting Canadian children and society by lowering S pneumoniae-associated disease. Additional gains from herd protection and further reductions in antimicrobial resistance will be achieved as more Canadian children younger than five years of age are routinely vaccinated with PCV7. PMID- 18418480 TI - A recurrent headache. PMID- 18418481 TI - Public reporting: Illumination or shadows? PMID- 18418482 TI - Central venous catheter-associated bloodstream infections in hemodialysis patients: Another patient safety bundle? PMID- 18418483 TI - Transfusion and risk of infection in Canada: Update 2006. PMID- 18418484 TI - Discharge Delay in Patients with Community-acquired Pneumonia Managed on a Critical Pathway. AB - INTRODUCTION: It has previously been reported that a critical pathway for community-acquired pneumonia (CAP) significantly reduces bed days per patient managed but results in no difference in average length of stay, suggesting that discharge criteria were not successfully implemented. The present study sought to identify factors in the timing of discharge not taken into account by discharge criteria. METHODS: Patients admitted with CAP and placed on a pneumonia critical pathway were studied. Patients' functional and cognitive status were evaluated using the Barthel Index, Hierarchical Assessment of Balance and Mobility (HABAM) and the Mini-Mental Status Examination. Once discharge criteria were met, the patient, a family member and the treating physician were interviewed to identify other factors contributing to length of stay. RESULTS: Thirty-one patients were enrolled in the study; 12 were discharged when they met discharge criteria and 19 stayed in hospital longer. There were no differences between patients discharged at stability versus those with an increased length of stay in terms of demographics, pneumonia severity score, functional or cognitive status at discharge using the Barthel Index (87.3+/-11.1 versus 83.8+/-8.6, respectively; P=0.46) and MMSE (27.1+/-1.1 versus 27.3+/-1.1, respectively; P=0.64); however, there was a significant difference in HABAM score at the time clinical stability was reached (22.6+/-1.3 versus 17.4+/-3.5, respectively; P=0.03), which correlated with physician and family assessments of patients' readiness for discharge. CONCLUSIONS: HABAM may be a useful tool to identify patients at risk of remaining in hospital after objective discharge criteria are met. Additional resources may be targeted at these patients to reduce length of stay in CAP. PMID- 18418485 TI - Escherichia Coli: From Genome Sequences to Consequences (or "Ceci n'est pas un elephant..."). AB - The present article summarizes a presentation given by Professor Mark Pallen of the School of Medicine at the University of Birmingham (Birmingham, United Kingdom) for the Fourth Stanier Lecture held in Regina, Saskatchewan, on November 9, 2004. Professor Pallen's lecture, entitled 'Escherichia coli: From genome sequences to consequences', provides a summary of the important discoveries of his team of research scientists in the area of genetic sequencing and variations in phenotypic expression. PMID- 18418486 TI - Rupture, invasion and inflammatory destruction of the intestinal barrier by Shigella: the yin and yang of innate immunity. AB - Shigella is a Gram-negative bacterial species of the family Enterobacteriaceae that causes bacillary dysentery in humans. This acute colitis reflects the capacity of the microorganism to disrupt, invade and cause the inflammatory destruction of the intestinal epithelium. The pathogenesis of the Shigella infection can be seen as a disruption of the homeostatic balance that protects the gut against inflammation in the presence of its commensal flora. This provides the unified view that enteroinvasive pathogens allow for the identification of key signalling molecules and pathways involved in the regulation of intestinal inflammation, and more generally, in the regulation of the innate and adaptive immune response. PMID- 18418487 TI - Acanthamoeba species Keratitis in a Soft Contact Lens Wearer Molecularly Linked to Well Water. AB - Acanthamoeba species keratitis has been associated with soft contact lens wear. In the present report, an epidemiological link was established between the patient's isolate and well water from the home using molecular methods. To the authors' knowledge, this is the first case in Canada where such a link has been established. Primary care practitioners and specialists, including ophthalmologists and infectious diseases specialists, must maintain a high degree of clinical suspicion in soft contact lens wearers with keratitis unresponsive to conventional topical and systemic treatment. PMID- 18418488 TI - First Canadian Reports of Cervical Adenitis Due to Mycobacterium Malmoense and a 10-year Review of Nontuberculous Mycobacterial Adenitis. AB - The present report reviews a decade of experience with nontuberculous mycobacterial adenitis at a pediatric referral centre, noting that patients are often subjected to multiple ineffective antibiotic courses, and that delays in diagnosis and referral for appropriate therapy are common. Notable clinical features include a mean age of presentation of 3.4 years, a male-to-female ratio of 1:1.5 and a gradual onset of painless, unilateral cervical adenopathy. Fever was absent in most patients (77%), and the disease failed to respond to antistaphylococcal antibiotics. The mean time to correct diagnosis was longer than three months (15 weeks). The clinical features of the disease are highlighted and presented with a practical diagnostic approach to the child with subacute/chronic adenitis. New molecular diagnostic tools and emerging mycobacteria are discussed, including the first reports of Mycobacterium malmoense adenitis in Canada. PMID- 18418489 TI - Vibrio vulnificus Septicemia After Handling Tilapia Species Fish: A Canadian Case Report and Review. AB - BACKGROUND: Vibrio vulnificus can cause a necrotizing soft tissue infection or primary septicemia; these infections are collectively known as vibriosis. This bacterium is commonly found within molluscan shellfish. Primary septicemia is often fatal, principally affecting persons with chronic liver disease. CASE PRESENTATION: A fatal case of V vulnificus sepsis that developed in a patient with chronic hepatitis B and chronic renal failure is reported. Diagnosis was made by isolation of the pathogen by blood culture. Upon further questioning, the patient's family recounted that the patient had handled and ingested Tilapia species fish in the hours preceding the patient's presentation. Despite treatment with doxycycline and cefotaxime, in conjunction with supportive care in the intensive care unit, the patient died on day 7 from multiple organ dysfunction. CONCLUSION: The present case highlights the need to consider V vulnificus in the microbiological differential diagnosis when a person presents with sepsis and bullous cutaneous lesions. The importance of educating patients with liver disease (and certain other chronic diseases) about the need to be cautious when handling or consuming seafood is underscored. PMID- 18418490 TI - A magician's tale. PMID- 18418491 TI - Two cases of miliary tuberculosis and elevated levels of cancer antigen 125. PMID- 18418492 TI - Physical Plant Design and Engineering Controls to Reduce Hospital-acquired Infections. PMID- 18418493 TI - Canadian Consensus Recommendations for the Optimal Use of Enfuvirtide in HIV/AIDS Patients. AB - BACKGROUND AND OBJECTIVES: An eight-member group consisting of Canadian infectious disease and immunology specialists and a family physician with significant experience in HIV management was convened to update existing recommendations, specifically intended for use by Canadian HIV-treating physicians, on the appropriate use of enfuvirtide in HIV/AIDS patients with resistance to other antiretroviral drugs. METHODS: Evidence from the literature and expert opinions of the group members formed the basis of the guidelines. Comments on the draft guidelines were obtained from other physicians across Canada with HIV expertise. The final guidelines represent the group's consensus agreement. RESULTS AND CONCLUSIONS: The recommendations were developed to guide physicians in optimal practices in patient selection for enfuvirtide treatment and subsequent patient management. The issues considered include positive predictors of response to enfuvirtide, stage of disease, optimization of the background regimen, early indicators of enfuvirtide response, and patient education and support. PMID- 18418494 TI - HIV Testing among Canadian Tuberculosis Cases from 1997 to 1998. AB - BACKGROUND: Recent evidence suggests a global rise in adult tuberculosis (TB) cases associated with HIV/AIDS. The World Health Organization, the United States Centers for Disease Control and Prevention, and the Public Health Agency of Canada advocate universal screening of all TB cases for HIV. The contribution of HIV to the TB burden in Canada remains unclear. METHODS: A retrospective cohort study was conducted of all TB cases reported in Canada from 1997 to 1998. The aim was to determine the proportion of patients that had an HIV test on record, and of these, the number of patients infected with HIV. RESULTS: From 1997 to 1998, 3767 TB cases were reported to the national TB surveillance system. In the present study, 3416 case records (90.7%) were included. The number of cases with a record of an HIV test was 736 (21.5%), and of these, 41.2% were tested within one month of TB diagnosis. Among the 703 cases with known HIV test results, the prevalence of HIV infection was 14.7%. Cases with an HIV test on record were more likely to have one or more risk factors for HIV, and also were more likely to be male, aged 15 to 49 years, of Aboriginal ethnicity, and to have smear-positive TB and both pulmonary and extrapulmonary disease at the time of diagnosis. DISCUSSION: These results suggest that HIV testing of TB patients is not universal, but rather selective, and is likely based on perceived risk factors for HIV as opposed to TB diagnosis alone. PMID- 18418495 TI - Central venous catheter-associated bloodstream infections occurring in Canadian intensive care units: A six-month cohort study. AB - OBJECTIVE: To determine the rate and risk factors associated with central venous catheter (CVC)-associated bloodstream infections (BSIs) in Canadian intensive care units (ICUs). DESIGN: A prospective, active six-month cohort with a nested case-control study. SETTING: Forty-one ICUs located in 19 Canadian hospitals. METHODS: Data were collected using a standardized format on all CVCs and patients when a CVC was inserted for more than 48 h. Results of microbiological studies and therapeutic interventions were recorded when a BSI occurred. RESULTS: There were 182 BSIs from 3696 CVC insertions in 2531 patients. Coagulase-negative staphylococci were responsible for 73% of the BSIs. Mean rates of CVC-associated BSIs per 1000 CVC days were 6.9, 6.8 and 5.0 in adult, neonatal and pediatric ICUs, respectively. Significant factors associated with BSI included duration of CVC insertion (OR=1.2, 95% CI 1.1 to 1.3), receiving total parenteral nutrition (OR=4.1, 95% CI 1.2 to 14.3) and having one or more CVCs (OR=3.1, 95% CI 1.5 to 6.5). In the case-control study, 80% of the variance in a backward elimination logistic regression analysis was explained by duration of CVC insertion (OR=1.2 per day), receiving chemotherapy (OR=6.1), more than one CVC insertion during the study (OR=3.5), insertion of a CVC with two or more lumens (OR=2.3), using the CVC to administer total parenteral nutrition (OR=1.6) and having a surgical wound other than a clean wound (OR=1.6). CONCLUSION: The present study identified risk factors explaining 80% of the variance associated with BSIs and is one of the largest reports on the rate of CVC-associated BSIs occurring in the ICU setting. PMID- 18418496 TI - An assessment of linezolid utilization in selected canadian provinces. AB - BACKGROUND: Linezolid is approved for the treatment of designated infections caused by methicillin-resistant and -susceptible Staphylococcus aureus and vancomycin-resistant Enterococcus faecium. OBJECTIVE: To characterize linezolid utilization since its launch in Canada in 2001. METHODS: Demographics, antimicrobial regimens, and clinical and resource utilization data for linezolid treated patients were collected retrospectively by hospital pharmacists at nine tertiary care hospitals in four provinces. Statistics describing linezolid utilization were calculated and the appropriateness of use was assessed according to a treatment algorithm based on recommendations of the Infectious Diseases Pharmacy Specialty Network in 2001. RESULTS: Ninety-nine linezolid courses were prescribed for 103 infections in 95 patients (mean age 57.8 years, 52.6% male) with an average length of hospital stay of 40.6 days. Fifty-three per cent of patients had an allergy to at least one antibiotic other than linezolid. The major use of linezolid was for treatment of skin and soft tissue infections (32.0%), followed by bacteremia (15.5%). The most prevalent pathogen was methicillin-resistant S aureus, identified in 44.7% of infections. Linezolid was primarily prescribed as the oral form following other intravenous anti-infectives (55.6% of courses) for an average duration of 14.4 days. The rate of appropriate utilization was 53% (range 25% to 75% by site). In 93.5% of courses deemed inappropriate, recommended first-line therapies were not attempted before linezolid. CONCLUSIONS: Linezolid was prescribed appropriately in approximately one-half of cases reviewed. The rate of appropriate utilization is similar to those rates reported in other Canadian antibiotic reviews. PMID- 18418497 TI - A Case of Acquired Rifampin Resistance in Mycobacterium Bovis Bacillus Calmette Guerin-induced Cystitis: Necessity for Treatment Guidelines. AB - A case of presumed bacillus Calmette-Guerin (BCG) cystitis in an elderly female patient following direct intravesical BCG instillation treatment for papillary transitional cell carcinoma is reported. The organism cultured from urine samples was eventually identified as a rifampin-resistant Mycobacterium bovis BCG isolate. Because the patient had received rifampin monotherapy during the course of treatment for presumed BCG disease, the clinical picture favoured acquired rifampin resistance. Sequencing of the target gene for rifampin (rpoB) confirmed a known mutation responsible for conferring high levels of resistance to both rifampin and rifabutin (Ser531Tyr). To the authors' knowledge, this is the first reported case of M bovis BCG disease in a non-HIV patient where the organism had acquired drug resistance to rifampin, and the second reported case of M bovis BCG that had acquired drug resistance. The present case demonstrates the necessity to re-evaluate appropriate guidelines for the effective treatment of BCG disease. PMID- 18418498 TI - An evaluation of extended incubation time with blind subculture of blood cultures in patients with suspected endocarditis. AB - BACKGROUND: In a small proportion of patients, bacterial endocarditis is due to organisms that grow slowly and may not be recovered in conventional blood cultures incubated for five days. This has led to recommendations for prolonged incubation and routine subculture of negative cultures. OBJECTIVE: The above mentioned approach is evaluated. METHOD: The microbiology of all blood cultures subjected to prolonged incubation and the charts of individuals who had organisms recovered after five days were evaluated to determine their clinical significance. RESULTS: In all, 507 blood cultures were handled using an extended incubation and blind subculture protocol. Fifty-three blood cultures in 27 patients were positive. Blood cultures were positive after five days in only five cases; patient outcomes were not affected by the results in any of these cases, although several fastidious organisms (ie, Haemophilus paraphrophilus and Haemophilus parainfluenzae) were recovered in the first five days of incubation. CONCLUSION: Prolonged incubation and blood subcultures in patients with suspected endocarditis or infections due to fastidious organisms do not represent a wise use of increasingly scarce resources. PMID- 18418499 TI - Prolonged cough in children: a summary of the Belgian primary care clinical guideline. AB - Prolonged cough is a frequent problem in the community. Several studies in the school setting have found that as many as 4.8% to 10.4% of children suffer from prolonged cough. There is no consensual definition of prolonged cough. In this guideline, we define prolonged cough as a daily cough lasting for more than three weeks. The literature review did not identify any quality study on the aetiology of prolonged cough in children in primary care. A diagnostic decision-tree based on the systematic literature review and expert opinion is proposed. Doctors should seek signs of any serious underlying condition. Chronic productive purulent cough should always be investigated. A careful evaluation of the impact of cough on the quality of life of the child is necessary. In absence of signs of specific underlying illness, coughing is generally a self-limiting condition. Symptomatic treatments have not yet been proven to be effective, and many of them may cause serious side effects. Their use should therefore be limited. PMID- 18418500 TI - UK Resuscitation Council guidelines on emergency treatment of anaphylactic reactions: a primary care perspective. PMID- 18418501 TI - Perceptions, impact and management of asthma in South Africa: a patient questionnaire study. AB - INTRODUCTION: A number of studies from around the world have indicated that asthma morbidity is still unacceptably high. In the AIRE study over one-third of children and half of the adults reported daytime symptoms at least once a week. This study was conducted to understand the impact (including the impact on health related quality of life) of asthma on South African asthmatics. MATERIALS AND METHODS: General Practitioners (GPs) and pharmacists in South Africa were randomly identified from the Medical Association database. These individuals were approached and asked to recruit asthmatic patients to complete a questionnaire. The questionnaire inquired about symptoms, quality of life, complications, trigger factors, associated allergic conditions, medication used, medication preference, medication adherence and concerns about the condition. RESULTS: 3347 respondents returned their demographic data but only 710 met the criteria for analysis, ie. had asthma and were presently on controller medication. Symptom analysis revealed that 21.4% of respondents were coughing on most days, 25.6% were wheezing on most days, and 22.8% were experiencing night-time symptoms on most days. Symptoms were exacerbated by exercise in 56.9%, while nocturnal wakening due to asthma occurred in 36.9% more than four times per week. Only 35.1% of respondents had not missed school or work in the preceding year. 45.4% of individuals worry about side effects of asthma therapy. CONCLUSION: This study indicates that there is under-treatment, inappropriate treatment and/or lack of patient education for asthma patients in South Africa. The data support the notion that poor therapy and/or poor patient adherence has an enormous impact on the health-related quality of life of South Africa's asthmatics. PMID- 18418502 TI - Different ways of looking at things. PMID- 18418503 TI - Triple classification of diagnosis of dry eyes. PMID- 18418504 TI - The dendritic cell in allergic conjunctivitis. AB - The acquired immune response in health and disease is initiated when foreign antigens are processed and presented to T lymphocytes via antigen-presenting cells as peptides in the context of Class I and II major histocompatibility complex antigens. It is now clear that there are various types of antigen presenting cells and that the phenotype of these cells (together with the milieu of the tissue or lymphoid organ) dictates the nature of the immune response to the antigen. Very little is known about the phenotype, distribution, and roles of dendritic cell subtypes that contribute to the pathophysiology of type I hypersensitivity reaction in the ocular surface. We review what has been learned from studies of both human ocular allergy and murine models and comment on how this compares to allergic reactions in other mucosal tissues. PMID- 18418505 TI - A new look at lubrication of the ocular surface: fluid mechanics behind the blinking eyelids. AB - The concept of the dacruon was presented by the author in this journal in July 2007. Dacruon, defined as "the body of unshed fluid, constantly occupying the ocular surface sac [OSS], comprising the mucoaqueous pool [MAP] and its covering lipid sealant," prompts a fresh consideration of OSS lubrication. The author notes scientific agreement that in the preocular, interpalpebral OSS (the menisco optical domain), the mucous gel of the MAP adheres to subjacent bulbar epithelium. In the retropalpebral recesses (the "lubrication domain"), lid and globe epithelia are juxtaposed. The author proposes that microvilli and glycocalyx "grasp" the bases of dacruon mucous gels, enabling lid and globe to "drive" fluid movement. The adherent gels and associated low viscosity mucous modules mitigate friction. There is no substantive lipid layer. The modules abut, "mirror-image" fashion, forming an interface concentric with the eyeball surface about midway between the palpebral and bulbar mucosae. Here, kinetic energy originating from both lid and globe has been progressively dissipated by fluid friction, residual movement reduced to creeping flow. Shear stress is displaced from the rapidly moving epithelia, to occur between the more remote, slow-moving midzone fluids, minimizing frictional blink-related microtrauma. The midway interface serves as a "slip interface," crucial to the mucoaqueous lubrication of the OSS. Concomitantly, the OSS also forms the anterior lubricating compartment of the "ball and socket" ocular joint. PMID- 18418506 TI - The ophthalmologic management of acute Stevens-Johnson syndrome. AB - Stevens-Johnson syndrome (SJS) and its more severe variant, toxic epidermal necrolysis (TEN), cause significant ocular surface damage. The majority of affected patients develop conjunctival inflammation during the acute phase of the disease. If intense, this inflammation yields permanent destruction of the normal mucosal tissue of the ocular surface and eyelids. Loss of the normal glandular structures leads to severe dry eye problems and vision loss. Medical treatments do little to arrest these problems, and repairing the damage after the acute phase is difficult, if not impossible. Recently, the application of cryopreserved amniotic membrane to the ocular surface during the acute phase of the disease has shown great promise in limiting the destructive inflammation and its long-term sequelae. A rationale for this treatment and a detailed description of the application procedure are provided, along with practical guidelines for the managing surgeon. PMID- 18418507 TI - John M. Tiffany, PhD. PMID- 18418508 TI - Conducting clinical trials: even more challenges. PMID- 18418509 TI - Acute myocardial infarction following transfusion reaction. AB - Indications for blood transfusion in critically-ill patients in general have become very restrictive. Still, the role for blood transfusion in the setting of acute coronary syndrome is unclear, as evidence regarding outcomes with different blood transfusion strategies has been conflicting. We report a 53-year-old Hispanic woman with acute ST-elevation myocardial infarction (STEMI) following a febrile nonhaemolytic transfusion reaction. Coronary angiogram showed complete occlusion of intracoronary stent in the left anterior descending (LAD) artery. Angiogram done earlier in the day had shown complete patency of the LAD stent with good distal flow. This suggests a possible role for the febrile nonhaemolytic transfusion reaction in triggering the stent occlusion and development of the STEMI. PMID- 18418510 TI - Afibrinogenaemia: a rare cause of young myocardial infarct. AB - Afibrinogenaemia is a quantitative deficiency of fibrinogen that is associated with an increased bleeding tendency. Patients with afibrinogenaemia are at risk of excessive bleeding, but unusually, these patients may also experience thrombotic episodes. We report a 27-year-old man, with a known history of afibrinogenaemia, and who presented with acute onset of chest pain. Clinical examination was normal. Electrocardiogram revealed features suggestive of inferior and right ventricular myocardial infarction. This diagnosis was confirmed on echocardiogram, and by the elevated levels of cardiac enzymes troponin T and Creatinine kinase-MB. In view of the existing afibrinogenaemia, thrombolytics and anticoagulants were not given. He was managed with antiplatelets, nitrates, statins and angiotensin converting enzyme inhibitors. Young coronary artery disease risk factors (including thrombophilia) were otherwise normal. The patient tolerated dual antiplatelet therapy without any bleeding. The thrombotic episode occurring in tandem with the afibrinogenaemia, as have been observed in previous reports, is likely to be the cause for acute myocardial infarction in our patient. PMID- 18418511 TI - 'Moya' than meets the eye: neurofibromatosis type 1 associated with Moyamoya syndrome. AB - Moyamoya syndrome (MMS) is an uncommon association of neurofibromatosis type 1 (NF1). We describe a seven-year-old chinese girl with NF1 and unilateral MMS with multiple hyperintensities on T2-weighted magnetic resonance (MR) images. The ischaemic lesions in the ipsilateral white matter were hypointense on fluid attenuated inversion recovery (FLAIR) MR images, in contrast to the hyperintense "unidentified bright objects" (UBOs) of NF1. Neuroradiologists should be aware of associated MMS in NF1 patients, and distinguish the effects of ischaemia from UBOs, especially on FLAIR MR imaging. PMID- 18418512 TI - Paradoxical vocal cord dysfunction: when a wheeze is not asthma. AB - Vocal cord dysfunction (VCD) is an uncommon condition which often mimics asthma in presentation and severity. We present nine- and 11-year-old female siblings with vocal cord dysfunction, which is a dysfunction of the larynx involving unintentional paradoxical adduction of the vocal cords during inspiration. We evaluated the use of exercise testing in conjunction with pulmonary function testing in suspected vocal cord dysfunction. Although normal pulmonology function tests were elicited with the patient at rest, exercise testing revealed blunting of the expiratory loop with attenuation of the inspiratory loop unique to VCD. The child underwent video laryngoscopy in the specialised voice clinic, which confirmed vocal cord dysfunction. Exercise testing is a rapid and noninvasive means of diagnosing vocal cord dysfunction in a small subset of patients, but video laryngoscopy, with training manoeuvres to elicit paradoxical vocal cord movements in VCD, remains the gold standard of diagnosis of VCD. PMID- 18418513 TI - Elastic cord-related ocular injury. AB - We report a case of an elastic cord-related ocular injury, to increase the public's awareness of the seriousness of this type of injury. A 43-year-old Chinese postman presented with decreased visual acuity in the left eye after being hit by an elastic cord at work. Slit lamp examination revealed pupillary sphincter rupture with iridodonesis and phacodonesis. There was also a traumatic nuclear cataract, gross hyphaema and vitreous haemorrhage. The patient was started on topical steroids and antibiotics. His visual acuity and vitreous haemorrhage improved during subsequent visits. Elastic cord-related ocular injuries have potential sight-threatening consequences. The public should be aware of the possible danger of using elastic cords. PMID- 18418514 TI - Isolated accessory limb of lower eyelid with multiple dermal appendages. AB - We report the first case in the English literature of an isolated occurrence of accessory limb with multiple dermal appendages in a ten-month-old boy. This condition presented at birth as a limb bud below the left eyelid, multiple dermal appendages in the adjacent part of the face below the left orbit and on the upper part of the face. No anomalies of the ocular structures or central nervous system were identified. Accessory limb with multiple dermal appendages, in the absence of a congenital cystic eye, is an extremely rare condition representing a benign aberration in the developing musculoskeletal system. We present the first of such a case and endeavour to explain the embryological basis behind it. PMID- 18418515 TI - Monocular blindness due to central retinal artery occlusion in bipolar hemireplacement arthroplasty of the hip. AB - Though rare, perioperative loss of vision after non-ocular surgeries is a disastrous complication. This has been reported after spine surgery or cardiopulmonary bypass surgery. We present an otherwise healthy 54-year-old man who underwent bipolar hemireplacement for a fractured neck of femur, and developed perioperative monocular visual loss due to central retinal artery occlusion, a complication hitherto unreported in hip replacement surgeries. The possible aetiological factors and the measures to prevent this complication are discussed. PMID- 18418516 TI - Two cases of deletion 5p syndrome: one with paternal involvement and another with atypical presentation. AB - We report two cases of deletion 5p or cri du chat syndrome (CdCS) with different presentations and risks of transmission: one case with paternal chromosome 5 involvement and another, a de novo case with atypical clinical presentation. Cytogenetic analysis was performed on the two cases and their parents. GTG-banded karyotype analysis of Cases 1 and 2 revealed abnormal 46,XY,del(5)(p13-15) male karyotypes. For Case 1, the mother showed normal female karyotype while the father showed an abnormal karyotype involving a balanced translocation 46,XY,t(5;10)(p13;p15). For Case 2, however, both parents showed a normal karyotype pattern. In Case 1, the clinical features, particularly the distinct facial phenotype in combination with a characteristic cat-like cry and hypotonia, aided in the diagnosis at birth and the karyotype analysis was resolutive. The boy in Case 2 presented with atypical clinical features. Even though this patient had multiple syndromic features, the typical high pitched cat-like cry was not prominent. Instead, the patient manifested persistent stridor (from day three of life), which might have prevented the clinician from suspecting CdCS at birth. However, when this patient was presented at seven months of age for cytogenetic analysis, a confirmatory diagnosis of CdCS was established. For children with congenital abnormalities, an early clinical diagnosis confirmed through cytogenetic and molecular investigations, is important for providing personalised diagnostic and prognostic evaluation, and also for genetic counselling on the reproductive risk, particularly for patients with parental chromosome translocation involvement. PMID- 18418517 TI - Common concerns regarding breastfeeding in a family practice setting. PMID- 18418518 TI - Industrial accident-related ocular emergencies in a tertiary hospital in Singapore. AB - INTRODUCTION: We present a review of industrial accident-related ocular trauma, from the perspective of the emergency setting of a tertiary hospital in Singapore. METHODS: A retrospective analysis of patients seen by Tan Tock Seng Hospital's emergency ophthalmology service over a six-month period was performed. Data on clinical presentation, cause of injury, use of protective eyewear and subsequent losses of days-of-work were collected via a standardised telephone interview and review of case-sheets. RESULTS: A total of 300 persons presented with a diagnosis of industrial accident-related ocular trauma, out of the 1,460 patients seen during the study period. 95.7 percent were non-residents and the average age was 31 years. 99.3 percent were males. 66.0 percent were provided with protective eyewear, while 44.7 percent of those were non-compliant. The most common types of injuries were superficial foreign body (71.3 percent), chemical injury (10 percent) and blunt trauma (4 percent), while severe injury requiring admission and emergency surgery occurred in 15 cases. Average loss of days of work was 3.4 days. CONCLUSION: Industrial accident-related ocular trauma comprises a relatively large proportion of the patients requiring ophthalmic review at the emergency service level in Singapore. These patients are mainly young, non-resident men and the injuries were generally minor. These are largely preventable with the use of well-fitting protective eyewear and strict compliance. This would greatly reduce the unnecessary loss of workdays. Therefore, there is a need to review the design, and reinforce the strict implementation of occupational eye safety programmes, especially among non resident workers. PMID- 18418519 TI - 22q11.2 deletion syndrome in Singapore (2000-2003): a case for active ascertainment. AB - INTRODUCTION: The 22q11 deletion syndrome (22q11DS) is associated with many congenital structural anomalies, notably cardiac defects (conotruncal anomalies) and velopharyngeal insufficiency, as well as neurodevelopmental and psychiatric findings in later life. Recent studies have tried to ascertain the true population incidence of this condition. However, this is difficult due to possible under-ascertainment from incomplete genetic testing in possible cases. The aim of this study is to investigate the local incidence and association of this deletion syndrome with other congenital structural anomalies, with emphasis on cardiac defects. METHODS: Data of 22q11 deletion cases born in 2000-2003 were retrieved from the Singapore National Birth Defects Registry (NBDR) and analysed. Data of congenital cardiac defect cases notified to NBDR in the same period were also retrieved and compared with the deletion cases. RESULTS: There were a total of 17 cases of 22q11DS in the four-year period 2000-2003, giving an overall incidence of 1.02 per 10,000 live-births or one in 9,804 births. 94 percent (16/17 cases) were associated with other structural anomalies, and of these, 68.8 percent (11/16 cases) had single system anomalies. Cardiac anomalies were the most common (100 percent). The deletion contributed to 0.86 percent (one in 116 cases) of all cardiac defects born during the same period. A higher contribution of this deletion was noted for interrupted aortic arch (10 percent), pulmonary atresia (12.7 percent) and truncus arteriosus (11.1 percent). CONCLUSION: In view of the high proportion of this deletion among certain cardiac defects, genetic testing should be made available to investigate the true burden and contribution of this deletion. As more genetic testing is done for this deletion, we are likely to see an increase in incidence, reflecting the true prevalence of this condition. PMID- 18418520 TI - Prevalence of malnutrition and its risk factors in stroke patients residing in an infirmary. AB - INTRODUCTION: The aim of this study was to investigate the prevalence of malnutrition, and its associated risk factors, in stroke patients residing in an infirmary in Hong Kong. METHODS: A cross-sectional retrospective study of 61 stroke patients residing in an infirmary was conducted. Baseline demographic data, including age, gender, smoking habit, and alcohol consumption, were obtained on admission. Nutritional status was assessed according to anthropometric parameters. Malnutrition was defined as having a body mass index (BMI) of below 18.5 kg per square metres for both gender and a serum albumin level of less than 35 g/L. 12 risk factors associated with malnutrition were evaluated according to established protocols. RESULTS: 61 of the 93 patients assessed had a history of cardiovascular accident and gave consent to participate in the study. Among them were 28 (46 percent) women and 33 (54 percent) men. The mean length of stay of these patients was 850 days (or 28 months). The mean age of these patients was 76 (standard deviation 12.8) years. Among the patients, five (8.2 percent) were malnourished and 56 (91.8 percent) were adequately nourished. There were no significant differences in the distribution of eight risk factors between the malnourished and nourished groups. These risk factors were a previous history of alcohol consumption, comorbidities (five or more), polypharmacy (five or more), diabetes mellitus, impaired functional status of daily living, impaired mobility (wheelchair- or bed-bound), tube-feeding, and edentulism. Insufficient data was available to assess the effects of two risk factors: depressed mood and impaired cognitive function. The distribution of another two risks factors (previous history of smoking and dysphagia) was significantly different between the malnourished and nourished groups. Odds ratios of smoking and dysphagia associated with malnourishment were approximately 3.3 and 2.6, respectively. CONCLUSION: Five of 61 (8.2 percent) stroke patients residing in an infirmary were malnourished. Two risk factors significantly associated with malnutrition were previous history of smoking and dysphagia. It is recommended that smoking history be elicited during routine history-taking of all stroke patients and particular nutritional attention be given to these at risk patients. It is also emphasised that the management of dysphagia should follow a standardised protocol and form an integral element of patient care. PMID- 18418521 TI - Blood pressure variability and arterial elasticity in hyperlipidaemic subjects. AB - INTRODUCTION: It is debatable whether the assessment of low density lipoprotein or total cholesterol (TC) alone is sufficient to identify an individual's risk of having myocardial infarction. In the Framingham study, the risk of coronary artery disease was better indicated by an increase in the TC to high density lipoprotein cholesterol (TC: HDL) ratio. The aim of this study is to determine the relationship between blood pressure variability (BPV) and arterial compliances in hyperlipidaemics, which was defined as TC:HDL of more than 5.0 as compared to normolipidaemics. METHODS: 22 subjects with hyperlipidaemia were age , gender- and weight-matched with normolipidaemic controls. 24-hour ambulatory blood pressure monitoring was recorded and arterial compliances were measured. RESULTS: There were significantly higher 24-hour systolic (SBP) (19.9 +/- 6.1 mmHg vs. 16.1 +/- 4.4 mmHg, p-value is less than 0.01), diastolic (16.6 +/- 4.7 mmHg vs. 13.9 +/- 4.8 mmHg, p-value is less than 0.05) and mean arterial (16.3 +/ 4.9 mmHg vs. 13.3 +/- 4.7 mmHg, p-value is less than 0.05) BPVs in the hyperlipidaemic group as compared to the normolipidaemic group. There were no significant differences in large and small arterial compliances between groups. There was a significant inverse relationship between SBP and large arterial compliance (r-value equals to -0.46, p-value is less than 0.05). There was no correlation between BPV and arterial compliances. CONCLUSION: The BPV was higher in hyperlipidaemic subjects as compared to normolipidaemic subjects. Large arterial compliance was negatively correlated with SBP in hyperlipidaemic subjects. PMID- 18418522 TI - Thrombolytic failure with streptokinase in acute myocardial infarction using electrocardiogram criteria. AB - INTRODUCTION: This study was primarily aimed to determine the failure rate of thrombolysis with streptokinase in acute myocardial infarction using electrocardiogram criteria and its association between various independent variables and outcome parameters. METHODS: A total of 192 subjects were recruited into this retrospective observational study. Thrombolysis failure with streptokinase was defined using electrocardiogram criteria of less than 50 percent reduction in ST elevation in the worst infarct lead. Multivariate analysis was used to test association with study outcome. RESULTS: A total of 109 patients (56.8 percent) failed thrombolysis using streptokinase. The failures were associated with five variables in multiple logistic regression analysis (backward stepwise method) including anterior location of myocardial infarct (odds-ratio [OR] 0.07, 95 percent confidence interval [CI] 0.03-0.16; p-value is less than 0.001), longer door-to-needle time (OR 1.01, 95 percent CI 1.00-1.02; p value is 0.02), diabetes mellitus (OR 3.13, 95 percent CI 1.13-8.69; p-value is 0.03), hypertension (OR 2.06, 95 percent CI 0.92-4.60; p-value is 0.08) and high total white cell count (OR 1.12, 95 percent CI 1.01-1.24; p-value is 0.03). Thrombolysis failure with streptokinase was associated with recurrent acute coronary syndrome (crude OR 2.49, 95 percent CI 1.16-5.32; p-value is 0.02) and death after one year (crude OR 7.61, 95 percent CI 0.95-61.24; p-value is 0.04). CONCLUSION: This study showed that streptokinase had a failure rate of 56.8 percent. History of diabetes mellitus, history of hypertension, anterior location of myocardial infarction, longer door-to-needle time and high total white cell count were highly predictive of thrombolysis failure using streptokinase. This group of patients may benefit from other early reperfusion strategy. PMID- 18418523 TI - Audit on cardiovascular disease preventive care in general practice. AB - INTRODUCTION: Cardiovascular disease is a major cause of morbidity and mortality. Primary care doctors as general practitioners (GPs) play a central role in prevention, as they are in contact with a large number of patients in the community through provision of first contact, comprehensive and continuing care. This study aims to assess the adequacy of cardiovascular disease preventive care in general practice through a medical audit. METHODS: Nine GPs in Malaysia did a retrospective audit on the records of patients, aged 45 years and above, who attended the clinics in June 2005. The adequacy of cardiovascular disease preventive care was assessed using agreed criteria and standards. RESULTS: Standards achieved included blood pressure recording (92.4 percent), blood sugar screening (72.7 percent) and attaining the latest blood pressure of equal or less than 140/90 mmHg in hypertensive patients (71.3 percent). Achieved standards ranged from 11.1 percent to 66.7 percent in the maintenance of hypertension and diabetic registries, recording of smoking status, height and weight, screening of lipid profile and attaining target blood sugar levels in diabetics. CONCLUSIONS: In the nine general practice clinics audited, targets were achieved in three out of ten indicators of cardiovascular preventive care. There were vast differences among individual clinics. PMID- 18418524 TI - Management of mesenteric vascular occlusion. AB - INTRODUCTION: The aim of this study was to evaluate our recent clinical management of mesenteric vascular occlusion (MVO) at Al-Ain Hospital, United Arab Emirates. METHODS: A retrospective study was performed including all patients who were diagnosed to have MVO from December 2001 to May 2005. The records were studied with regard to clinical features, risk factors, diagnosis, treatment, and outcome. RESULTS: Of the 14 patients studied, seven patients experienced mesenteric venous thrombosis (MVT), five patients mesenteric arterial occlusion (MAO), and two patients were found to have both MVT and MAO. The main risk factor for MAO was ischaemic heart disease with atrial fibrillation in four patients (80 percent). No predisposing factors were identified in three patients with MVT (primary MVT 43 percent). Contrast-enhanced computed tomography was performed in all patients and was diagnostic in 12 (86 percent) patients. Seven patients (50 percent) underwent surgery. One patient died on the ninth postoperative day (overall mortality rate 7 percent). Seven patients (50 percent) were successfully managed conservatively, five of them had only MVT, one had combined MVT and MAO, and one had only MAO. CONCLUSION: Early diagnosis and prompt initiation of anticoagulation therapy, with operative intervention when indicated, are essential for a favourable outcome. PMID- 18418525 TI - Impact of Essentiale L on ethanol-induced changes in rat brain and erythrocytes. AB - INTRODUCTION: The present study was designed to investigate the effect of Essentiale L, a mixture of polyenylphospholipids from soybeans, on oxidative stress in various brain regions, on erythrocytes (RBC) and on RBC membrane composition in ethanol-administered rats. METHODS: Adult male albino rats of body weight 150-170 g were divided into four groups and administered either isocaloric glucose (5 g/kg body weight/day) or ethanol (6 g/kg body weight/day) through oral gavage. Essentiale L was administered to a set of ethanol-fed rats and the control rats at a dosage of 300 mg/kg body weight/day through oral gavage. The treatment protocol was carried out for 45 days. At the end of the experimental period, the animals were sacrificed, and the biochemical parameters related to the lipid profile, oxidative stress and thiol status were assayed in the brain regions, RBC and RBC membrane. RESULTS: Ethanol administration resulted in increased levels of lipid peroxidation products in RBC and different brain regions, such as the cortex, cerebellum, striatum, hippocampus and hypothalamus, and depletion of enzymatic and nonenzymatic antioxidants and alterations in oxidised glutathione/glutathione (GSSG/GSH) ratio and thiol groups (protein-bound and total), signifying oxidative stress. Ethanol-treated rats also showed significant alterations in protein content and lipid composition in RBC membranes. Significant differences in the relative proportions of hexose, hexosamine and sialic acid of the membranes were observed. Administration of Essentiale L prevented all the alterations induced by ethanol and returned their levels to near-normal. CONCLUSION: These findings suggest that Essentiale L, a therapeutic adjunct for liver diseases, also has bioprotective effects on nonhepatic tissues and cells. PMID- 18418526 TI - Should clinical normality be examined in medical course? AB - INTRODUCTION: In medical practice, some patients consult doctors for reassurance of normality, e.g. patients with throat discomfort. Therefore, medical graduates should be competent in diagnosing clinical normality. One way to assess clinical competence is by the objective structured clinical examination (OSCE). METHODS: In 2002-2006, five batches of medical students who completed their otorhinolaryngology posting in Universiti Malaysia Sarawak were examined with the same OSCE question on clinically normal vocal cords. There were five subquestions concerning structures, clinical features, diagnosis and management. All students had prior slide show sessions regarding normal and abnormal laryngeal conditions. RESULTS: The total number of students in 2002, 2003, 2004, 2005 and 2006 was 25, 41, 20, 30 and 16, respectively, and 100 percent responded. The average percentage of students with correct answers was 19.4, 2.4, 2.2, 21.2, and 2.4, in the subquestions 0.1 to 0.5, respectively, leaving the remaining relatively larger percentages with incorrect answers of various clinical abnormalities. A reason for these findings is examination fever by the students, who also assumed that all the stations had clinical abnormalities and required differentiating abnormalities from abnormalities, and not from normality. Without clinical normality OSCE questions, the assessment of the undergraduates' clinical competence in real life would seem incomplete. CONCLUSION: This study showed that a significantly large percentage of students answered incorrectly in the clinical normality OSCE. This may mean that more clinical normality OSCE questions should be included in the undergraduate medical examination to help undergraduates practise the need to look for, and become competent in, clinical normality in real life. PMID- 18418527 TI - Expression of Galectin-3 and Galectin-7 in thyroid malignancy as potential diagnostic indicators. AB - INTRODUCTION: It has been suggested that Galectin-3 (Gal-3) and Galectin-7 (Gal 7) are potential tumour markers for differentiating thyroid carcinoma from its benign counter part. Galectins are beta-galactoside-binding proteins with Gal-3 being a redundant pre-mRNA splicing factor. They are supposed to be p53-related regulators in cell growth and apoptosis, being either anti-apoptotic or pro apoptotic. Although the value of Gal-3 has been studied extensively, there is little knowledge regarding the expression of Gal-7 in thyroid malignancy. METHODS: We initiated an immunohistochemical (IHC) study on the expression of Gal 3 and Gal-7 on various thyroid lesions. Formalin-fixed paraffin embedded thyroid tissues were stained for IHC expression of Gal-3 and Gal-7 using monoclonal anti human Gal-3 antibody and anti-human Gal-7 antibody (R&D Systems Inc, MN, USA). Gal-3 and Gal-7 expressions were measured semiquantitatively on their distribution and staining intensity. RESULTS: A total of 95 cases were collected, including 32 benign and 63 malignant thyroid lesions. These contained 37 cases of papillary thyroid carcinoma, nine cases of papillary thyroid carcinoma follicular variant, 16 cases of follicular carcinoma, one case of anaplastic carcinoma, 14 cases of follicular adenomas and 18 cases of nodular goitre. Gal-3 expression was significantly strong in cancer cases compared to non-cancer cases (p-value is 0.000), while no significant difference was noted with Gal-7 expression (p-value is 0.870). CONCLUSION: Our findings suggested that the IHC localisation of Gal-3 is a useful marker in conjunction with routine haematoylin and eosin staining in differentiating benign from malignant thyroid lesions, while there is no significant adjunct diagnostic value in Gal-7 for thyroid malignancy. PMID- 18418528 TI - Patients' perception of transnasal gastroscopy. AB - INTRODUCTION: Gastroscopy is an excellent technique for investigation of upper gastrointestinal pathology. However, patients frequently request for conscious sedation as the procedure causes pain, pharyngeal reflex and nausea. Administration of conscious sedation incurs additional medical expenditure and risks. Transnasal gastroscopy, which became commercially available in 2005, does not induce pharyngeal reflex. The aim of this study was to examine patients' perception and satisfaction with transnasal gastroscopy performed in an office setting. METHODS: Questionnaires were administered to consecutive patients after the performance of transnasal gastroscopy. Patients received topical ten percent xylocaine spray to the nasal and pharyngeal cavities 1-2 minutes before the procedure. The transnasal endoscope used was a narrow upper gastrointestinal endoscope (EG270N5 [Fujinon, Saitama City, Japan]). RESULTS: Transnasal gastroscopy was attempted in 96 patients. The procedure failed in one patient due to a narrow nasal passage and had to be converted to oral route of intubation. Questionnaires were completed by 96 patients. There were 52 males (54 percent) with the median age of 43 (range 11-85) years. None of them received conscious sedation. 53 patients (56 percent) reported that there was no pain/discomfort during the procedure. On the Lickert visual analogue scale for pain from 0 to 10 points, 91 patients (96 percent) reported severity of pain of below 5 points. 85 patients (89 percent) reported they were satisfied or more than satisfied with the procedure. 84 patients (88 percent) were happy to undergo similar repeat procedure without sedation. 25 patients (26 percent) had undergone previous oral gastroscopy; 22 of these patients reported that transnasal route was definitely more comfortable than the oral route. There were two incidents of nosebleed which were self-limiting. CONCLUSION: Transnasal gastroscopy with a thin endoscope was found to be safe. The procedure is well-tolerated by patients without conscious sedation. Patients reported better preference for transnasal endoscopes in comparison to conventional transoral gastroscopy. PMID- 18418529 TI - Aeromonas hydrophila bacteraemia and portal pyaemia. AB - The Aeromonas species uncommonly cause disease in humans. We report portal pyaemia secondary to Aeromonas hydrophila bacteraemia occurring in a 71-year-old Chinese man with no history of hepatobiliary disease or malignancy. He presented with fever, rigors and abdominal bloating for four days and was subsequently found to have Aeromonas hydrophila bacteraemia, portal vein thrombosis and a psoas abscess. He was treated with ciprofloxacin and had a good recovery. Aeromonas hydrophila infection is an uncommon cause of intestinal and extraintestinal infection in man, but must be suspected in immunocompromised hosts and in those exposed to brackish or salt water. PMID- 18418530 TI - Gonadal mosaicism 45,X/46,X,psu dic(Y)(q11.2) resulting in a Turner phenotype with mixed gonadal dysgenesis. AB - A two-year-and-eight-month-old girl presented with clitoromegaly and short stature. Two cell lines, 45,X and 46,X,idic(Y)(q11.2), were observed. Cytogenetic and fluorescence in situ hybridisation investigations were carried out on her peripheral lymphocytes and gonadal cells, to determine the genotype-phenotype effect with respect to differential tissue distribution, effects of the sex determining region of the Y chromosome, and the break-points in the azoospermia factor region. PMID- 18418531 TI - Computed tomography of blunt abdominal trauma in children. AB - Computed tomography (CT) plays a major role in diagnosis of blunt abdominal trauma of haemodynamically-stable children. The purpose of this article is to review the CT findings in children with hepatic, splenic, renal, adrenal, pancreatic, bowel, and mesentery injuries and in children with blunt abdominal trauma and active haemorrhage. PMID- 18418533 TI - Traumatic complications of acupuncture. PMID- 18418534 TI - PharmedOut. PMID- 18418535 TI - Hypothalamic hamartoma: MR imaging and MR spectroscopic features. PMID- 18418537 TI - Severe bleeding from internal obturator muscle following tension-free vaginal tape Secur hammock approach procedure. AB - Various perioperative and postoperative complications have been reported after the use of pubo-vaginal tension-free vaginal tapes. The transobturator approach was introduced to minimize the potential complications connected with entering the retropubic space, but different types of complications have also been described. The next generation of recently introduced minitapes such as TVT-Secur is intended to minimize the incidence of complications. This report concerns a patient who underwent a tension-free vaginal tape procedure-TVT-Secur hammock approach-and developed severe bleeding from the internal obturator muscle, which required surgical intervention. PMID- 18418538 TI - The effects of zoledronic acid on serum lipids in multiple myeloma patients. AB - Nitrogen-containing bisphosphonates (N-BPs) inhibit osteoclast-mediated bone resorption and are widely used for tumor-associated osteolysis. The mechanism of action of these drugs has not been completely clarified, but it has been observed that N-BPs may inhibit squalene synthase or farnesyl pyrophosphate synthase. Zoledronic acid (ZA) represents a novel N-BP which also has antitumor activity. To explore the effects of ZA on serum lipids, we studied 26 patients with smoldering myeloma at diagnosis. Sixteen patients were treated with ZA (4 mg) at baseline and at months 1, 2, 4, and 6. The remaining 10 served as controls. In all subjects, total cholesterol (TC), triglycerides (TGs), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), and C-terminal telopeptide of type I collagen (CTX) were measured at baseline and after 1, 3, and 6 months. In treated patients, we observed a progressive and significant reduction of TC, with a maximum decrease of 13% at 6 months. Moreover LDL-C decreased by 21% at 6 months, while no significant difference was appreciated in HDL-C and TGs. Also, the indexes of cardiovascular risk improved after ZA administration: TC/HDL-C ratio progressively decreased by 17% and HDL C/LDL-C ratio increased by 36%, showing an effect that appears to be cumulative. In conclusion, ZA given intravenously at high doses in patients with smoldering myeloma seems to be able to modify the lipid profile with an improvement of atherosclerotic risk index. PMID- 18418539 TI - Block of persistent late Na+ currents by antidepressant sertraline and paroxetine. AB - Antidepressants, such as traditional tricyclic antidepressants (TCAs), are the first-line treatment for various pain syndromes. Available evidence indicates that TCAs may target Na+ channels for their analgesic action. In this report, we examined the effects of contemporary antidepressants sertraline and paroxetine on (1) neuronal Na+ channels expressed in GH3 cells and (2) muscle rNav1.4 Na+ channels heterologously expressed in Hek293t cells. Our results showed that both antidepressants blocked Na+ channels in a highly state-dependent manner. The 50% inhibitory concentrations (IC50) for sertraline and paroxetine ranged approximately 18-28 microM: for resting block and approximately 2-8 microM: for inactivated block of neuronal and rNav1.4 Na+ channels. Surprisingly, the IC50 values for both drugs were about 0.6-0.7 microM: for the open channel block of persistent late Na+ currents generated through inactivation-deficient rNav1.4 mutant Na+ channels. For comparison, the open channel block in neuronal hNav1.7 counterparts yielded IC50 values around 0.3-0.4 microM: for both drugs. Receptor mapping using fast inactivation-deficient rNav1.4-F1579A/K mutants with reduced affinities toward local anesthetics (LAs) and TCAs indicated that the F1579 residue is not involved in the binding of sertraline and paroxetine. Thus, sertraline and paroxetine are potent open channel blockers that target persistent late Na+ currents preferentially, but their block is not mediated via the phenylalanine residue at the known LA/TCA receptor site. PMID- 18418540 TI - Single-channel characterization of the rabbit recombinant RyR2 reveals a novel inactivation property of physiological concentrations of ATP. AB - Ryanodine receptor 2 (RyR2) cDNA has been available for more than 15 years; however, due to the complex nature of ligand gating in this channel, many aspects of recombinant RyR2 function have been unresearched. We established a stable, inducible HEK 293 cell line expressing full-length rabbit RyR2 cDNA and assessed the single-channel properties of the recombinant RyR2, with particular reference to ligand regulation with Ca2+ as the permeant ion. We found that the single channel conductances of recombinant RyR2 and RyR2 isolated from cardiac muscle are essentially identical, as is irreversible modification by ryanodine. Although it is known that RyR2 expressed in HEK 293 cells is not associated with FKBP12.6, we demonstrate that these channels do not exhibit any discernable disorganized gating characteristics or subconductance states. We also show that the gating of recombinant RyR2 is indistinguishable from that of channels isolated from cardiac muscle when activated by cytosolic Ca2+, caffeine or suramin. The mechanisms underlying ATP activation are also similar; however, the experiments highlighted a novel effect of ATP at physiologically relevant concentrations of 5-10 mM. With Ca2+ as permeant ion, 5-10 mM ATP consistently inactivated recombinant channels (15/16 experiments). Such inactivation was rarely observed with native RyR2 isolated from cardiac muscle (1 in 16 experiments). However, if the channels were purified, inactivation by ATP was then revealed in all experiments. This action of ATP may be relevant for inactivation of sarcoplasmic reticulum Ca2+ release during cardiac excitation-contraction coupling or may represent unnatural behavior that is revealed when RyR2 is purified or expressed in noncardiac systems. PMID- 18418541 TI - Epithelial barrier modulation by a channel forming peptide. AB - NC-1059 is a synthetic channel-forming peptide that provides for ion transport across, and transiently reduces the barrier integrity of, cultured epithelial monolayers derived from canine kidney (MDCK cells). Experiments were conducted to determine whether epithelial cells derived from other sources were similarly affected. Epithelial cells derived from human intestine (T-84), airway (Calu-3), porcine intestine (IPEC-J2) and reproductive duct (PVD9902) were grown on permeable supports. Basal short circuit current (Isc) was <3 microA cm(-2) for T 84, IPEC-J2 and PVD9902 cell monolayers and <8 microA cm(-2) for Calu-3 cells. Apical NC-1059 exposure caused, in all cell types, an increase in Isc to >15 microA cm(-2), indicative of net anion secretion or cation absorption, which was followed by an increase in transepithelial conductance (in mS cm(-2): T-84, 1.6 to 62; PVD9902, 0.2 to 51; IPEC-J2, 0.3 to 26; Calu-3, 2.3 to 13). These results are consistent with the peptide affecting transcellular ion movement, with a likely effect also on the paracellular route. NC-1059 exposure increased dextran permeation when compared to basal permeation, which documents an effect on the paracellular pathway. In order to evaluate membrane ion channels, experiments were conducted to study the dose dependence and stability of the NC-1059-induced membrane conductance in Xenopus laevis oocytes. NC-1059 induced a dose-dependent increase in oocyte membrane conductance that remained stable for greater than 2 h. The results demonstrate that NC-1059 increases transcellular conductance and paracellular permeation in a wide range of epithelia. These effects might be exploited to promote drug delivery across barrier epithelia. PMID- 18418542 TI - Characterisation of the components of the thioredoxin system in the archaeon Sulfolobus solfataricus. AB - The thioredoxin system is a redox machinery widely distributed in nature and involved in several cellular functions. It is constituted of thioredoxin reductase (Trx-B), its protein substrate thioredoxin (Trx-A) and NADPH. We have previously characterised a Trx-B from the hyperthermophile Sulfolobus solfataricus (SsTrx-B3) (Ruocco et al. in Biochimie 86:883-892, 2004). As in the genome of this archaeon, the gene coding for another Trx-B (SsTrx-B2) and for two Trx-A (SsTrx-A1, SsTrx-A2) have been putatively identified, these proteins were obtained as recombinant forms and characterised. SsTrx-B2, different from SsTrx B3, did not elicit a thioredoxin reductase activity. S. solfataricus possessed only one Trx-B (SsTrx-B3), which had two thioredoxins (SsTrx-A1 and SsTrx-A2) as substrates. These latter showed a homodimeric structure and catalysed insulin reduction using either DTT or NADPH/SsTrx-B3 as electron donors. In addition, the electron transfer between SsTrx-B3 and either SsTrx-A1 or SsTrx-A2 was fully reversible, thus allowing the determination of the redox potential of the thioredoxin system in S. solfataricus. Among the two thioredoxins, SsTrx-A2 appeared slightly more active and stable than SsTrx-A1. These data, besides shedding light on thioredoxin system in S. solfataricus, will contribute to add further information on this key enzyme system in Archaea. PMID- 18418543 TI - Archaeal diversity in a Fe-As rich acid mine drainage at Carnoules (France). AB - The acid waters (pH=2.73-3.4) that originate from the Carnoules mine tailings (France) are known for their very high concentrations of As (up to 10,000 mg l( 1)) and Fe (up to 20,000 mg l(-1)). To analyze the composition of the archaeal community, (their temporal variation inside the tailing and spatial variations all along the Reigous Creek, which drains the site), seven 16S rRNA gene libraries were constructed. Clone analysis revealed that all the sequences were affiliated to the phylum Euryarchaeota, while Crenarchaeota were not represented. The study showed that the structure of the archaeal community of the aquifer of the tailing stock is different to that of the Reigous Creek. Irrespective of the time of sampling, the most abundant sequences found inside the tailing stock were related to Ferroplasma acidiphilum, an acidophilic and ferrous-iron oxidizing Archaea well known for its role in bioleaching. Inversely, in Reigous Creek, a sequence affiliated to the uncultured Thermoplasmatales archaeon, clone YAC1, was largely dominant. This study provides a better understanding of the microbial community associated with an acid mine drainage rich in arsenic. PMID- 18418544 TI - Vertical distribution of bacterial and archaeal communities along discrete layers of a deep-sea cold sediment sample at the East Pacific Rise (approximately 13 degrees N). AB - The community structure and vertical distribution of prokaryotes in a deep-sea (ca. 3,191 m) cold sediment sample (ca. 43 cm long) collected at the East Pacific Rise (EPR) approximately 13 degrees N were studied with 16SrDNA-based molecular analyses. Total community DNA was extracted from each of four discrete layers EPRDS-1, -2, -3 and -4 (from top to bottom) and 16S rDNA were amplified by PCR. Cluster analysis of DGGE profiles revealed that the bacterial communities shifted sharply between EPRDS-1 and EPRDS-2 in similarity coefficient at merely 49%. Twenty-three sequences retrieved from DGGE bands fell into 11 groups based on BLAST and bootstrap analysis. The dominant groups in the bacterial communities were Chloroflexi, Gamma proteobacteria, Actinobacterium and unidentified bacteria, with their corresponding percentages varying along discrete layers. Pairwise Fst (F-statistics) values between the archaeal clone libraries indicated that the archaeal communities changed distinctly between EPRDS-2 and EPRDS-3. Sequences from the archaeal libraries were divided to eight groups. Crenarchaea Marine Group I (MGI) was prevalent in EPRDS-1 at 83%, while Uncultured Crenarchaea group II B (UCII B) abounded in EPRDS-4 at 61%. Our results revealed that the vertically stratified distribution of prokaryotic communities might be in response to the geochemical settings and suggested that the sampling area was influenced by hydrothermalism. The copresence of members related to hydrothermalism and cold deep-sea environments in the microbial community indicated that the area might be a transitional region from hydrothermal vents to cold deep-sea sediments. PMID- 18418545 TI - Adhesion of binary giant vesicles containing negative spontaneous curvature lipids induced by phase separation. AB - We report the adhesion of binary giant vesicles composed of two types of phospholipids, one has negative spontaneous curvature which tends to bend toward the head group and the other has zero spontaneous curvature. In a homogeneous one phase region, the giant vesicles do not adhere to each other, whereas in a coexisting two-phase region, the giant vesicles show adhesion. A fluorescence microscope observation reveals that the adhesion takes place through the domains rich in phospholipids having negative spontaneous curvature. We propose a phase separation induced hemifusion model where two apposed monolayers of adjacent vesicles are hemifused in order to reduce the bending energy of monolayers with negative spontaneous curvature and the boundary energy between the domains and matrix. We provide a strong evidence for the hemifusion model by lipid transfer experiments. PMID- 18418546 TI - Structure and rheological properties of model microemulsion networks filled with nanoparticles. AB - Model microemulsion networks of oil droplets stabilized by non-ionic surfactant and telechelic polymer C18 -PEO(10k)- C18 have been studied for two droplet-to polymer size ratios. The rheological properties of the networks have been measured as a function of network connectivity and can be described in terms of simple percolation laws. The network structure has been characterised by Small Angle Neutron Scattering (SANS). A Reverse Monte Carlo (RMC) approach is used to demonstrate the interplay of attraction and repulsion induced by the copolymer. These model networks are then used as matrix for the incorporation of silica nanoparticles (R = 10 nm), individual dispersion being checked by scattering. A strong impact on the rheological properties is found for silica volume fractions up to 9%. q(A-1). PMID- 18418548 TI - Cluster-like headache and idiopathic intracranial hypertension: a case report. AB - Cluster headache (CH) is a well-defined primary headache syndrome, but cases of symptomatic headache with clinical features of CH have been previously reported. Idiopathic Intracranial Hypertension (IIH) is a secondary headache disorder characterized by headache and visual symptoms, without clinical, radiological or laboratory evidence of intracranial pathology. Both papilloedema and IIH-related headache are typically bilateral, however asymmetrical or even unilateral localizations are described in literature. We report the case of a previously headache-free woman who presented cluster-like headache and asymmetrical papilloedema related to IIH. In our opinion the asymmetrical presentation supports, in this case, the hypothesis of cavernous sinus involvement in the IIH related cluster-like headache pathogenesis. PMID- 18418547 TI - Impact of headache in Europe: a review for the Eurolight project. AB - A recent health economic survey in Europe has suggested that migraine is the costliest among the neurological disorders. According to many studies, migraine and other disorders lead to widespread suffering, reduction of quality of life, and marked impairment of participation, both in work and social activities. The present literature survey was made in order to summarize what is known on the subject, as a preparation for a EU-supported study to assess the impact in several EU countries with similar methodology and the same research instrument. Previous studies have yielded relatively reliable data only for migraine, whereas the impact of tension-type headache is virtually unknown or only very incompletely known for most dimensions of headache impact. Some data do suggest, however, that this headache may be as important from a health economic and a public health perspective as migraine. In future studies it is important to get population-based data from various countries relevant for estimation of indirect (mostly absenteeism from work and reduced working efficiency when having headache) and direct costs (related to medication, consultations, investigations and hospitalisations). Also, the impact on ability to get education and participate in the workforce is very relevant, as is the impact on love life and family planning. The quality of life of headache patients should be measured by validated instruments. To get a complete picture, one should also ask about the effect on the life of partners and children, and on the possible impact even when headache-free (e.g. fear of the next attack). PMID- 18418549 TI - Possible iatrogenic bilateral cerebral ischemic infarcts in a woman with vasculitis. PMID- 18418550 TI - An atypical case of typical chest pain. PMID- 18418551 TI - Evaluation of end-tidal carbon dioxide role in predicting elevated SOFA scores and lactic acidosis. AB - The development of organ dysfunction is a key contributor to morbidity and mortality in sepsis. End-tidal carbon dioxide levels measured by non-invasive end tidal capnography (ETCO2) may provide a rapid assessment of a patient's underlying metabolic status. The objective of this study was to explore the association between ETCO2 and (1) organ dysfunction [sequential organ failure assessment (SOFA) score], and (2) serum lactate levels in febrile emergency department (ED) patients. Prospective, observational cohort study of a convenience sample of 97 adult (age 18 years or older) patients presented to an academic urban ED with a fever and suspected infection. The outcomes were ED SOFA score and serum lactate level. Based on prior studies, we categorized an ETCO2 <35 mmHg, a priori, as abnormal for the exposure. We defined clinically significant organ failure as a SOFA score of >2, and an abnormal lactate as >4 mmol/L. The correlation of ETCO2 with SOFA and lactate level was analyzed using Pearson correlation coefficient. Operating characteristics were calculated with 95% confidence intervals, along with the area under the curve (AUC). Among 97 patients enrolled, 5 (5%) had an abnormal lactate and 34 (35%) had a SOFA score >2. A significant correlation was found between ETCO2 and SOFA score (r = -0.35, p < 0.01), and ETCO2 and lactate level (r = -0.35, p < 0.01). A receiver operator curve for ETCO2 and SOFA >2 had an AUC of 0.69. ETCO2 of <35 has a sensitivity of 0.73 (95% CI 0.56-0.85) and specificity 0.50 (0.38-0.62) in predicting SOFA scores >2. ETCO2 <35 has a sensitivity of 0.60 (0.22-0.88) and specificity 0.42 (0.32-0.52) in predicting lactate >4 with an AUC of 0.62. We found a small, but statistically significant correlation, between ETCO2 and SOFA scores; however, based on questionable operating characteristics, the test seems to have limited ability to meaningfully impact clinical decision making. Larger confirmatory studies are required before final assessment. PMID- 18418552 TI - Expression of cell adhesion proteins and proteins related to angiogenesis and fatty acid metabolism in benign, atypical, and anaplastic meningiomas. AB - Most meningiomas are benign tumours of arachnoidal origin, although a small number have high proliferative rates and invasive properties which complicate complete surgical resection and are associated with increased recurrence rates. Few prognostic indicators exist for meningiomas and further research is necessary to identify factors that influence tumour invasion, oedema and recurrence. Paraffin sections from 25 intracranial meningiomas were analysed for expression of the proteins vascular endothelial growth factor (VEGF), VEGF receptors Flt1 and Flk1, E-cadherin, metalloproteinases 2 and 9 (MMP2, MMP9), CD44, receptor for hyaluronic acid-mediated motility (RHAMM), hyaluronic acid (HA), CD45, cyclooxygenase 2 (COX2), brain fatty acid binding protein (BFABP), Ki67, and proliferating cell nuclear antigen (PCNA). Correlations among protein expression were found for several markers of proliferation (Ki67, PCNA, MI) and microvessel density (MVD). COX2 expression increased with increasing with tumour grade and correlated with Ki67, PCNA, MI, MVD, and BFABP. BFABP expression also correlated with Ki67 and PCNA expression. Relationships were also identified among angiogenic factors (VEGF, Flt1, Flk1) and proliferation markers. Oedema was found to correlate with MMP9 expression and MMP9 also correlated with proliferation markers. No correlations were found for MMP2, E-cadherin, or CD44 in meningiomas. In conclusion Ki67, PCNA, MI, MVD, BFABP, and COX2 were significantly correlated with meningioma tumour grade and with each other. These findings, by correlating both intracellular fatty acid transport and eicosanoid metabolism with tumour proliferation, as determined by Ki67 labelling and mitotic index, suggest fatty acids are involved in the progression of meningiomas. PMID- 18418553 TI - Intensified monitoring of circadian blood pressure and heart rate before and after intravitreous injection of bevacizumab: preliminary findings of a pilot study. AB - BACKGROUND: A dose-dependent increase in arterial blood pressure (BP) was seen during bevacizumab treatment given intravenously for metastatic carcinoma. Because low systemic levels can also be expected after the intravitreal administration of bevacizumab, we looked for possible haemodynamic reactions of patients at higher risk of developing cardiovascular events after bevacizumab injection. METHODS: Ambulatory BP was monitored in 14 hypertensive patients receiving 1.25 mg intraocular bevacizumab for either choroidal neovascularization (CNV) or retinal proliferation associated with central retinal vein occlusion (CRVO). Circadian measurement was carried out twice, first at least 24 h prior to injection and second 72 h afterwards. Baseline evaluation before injection was compared with values taken in a matched control group. Taking a small random sample of two patients, serum concentration of bevacizumab and VEGF-A was measured at several time points. RESULTS: High incidence of pathologic BP values was found in the pre-injection measurement, even under anti-hypertensive treatment of the patients with CNV or CRVO. No general increase in BP was seen after the intravitreal injection (P = 0.01), although significantly reduced nocturnal dipping occurred as compared to before the injection (P = 0.006). Individual patients showed a rise in BP load subsequent to injection. A decline in serum VEGF-A was found to correspond to measureable levels of serum bevacizumab (up to 90 ng/ml). CONCLUSIONS: Before the intravitreal injection, BP values were increased in the majority of the patients. The elevated BP load might be related to probable pre-injection stressors. There seems to be no general rise in mean BP, heart rate and pulse pressure after intravitreal bevacizumab, although a decrease in serum VEGF-A can occur in individual patients. The reduced nocturnal dipping could be caused by pharmacodynamic effects on the vasal tone; this preliminary but striking finding warrants further investigation. PMID- 18418554 TI - Reversed papilledema in an MPS VI patient with galsulfase (Naglazyme) therapy. AB - MPS VI (mucopolysaccharidosis VI, known as Maroteaux-Lamy syndrome) is a multi systemic inherited disease, resulting from a deficiency of N-acetylgalactosamine 4-sulfatase, causing accumulation of the glycosaminoglycan (GAG) dermatan sulfate in all tissues. It is one of almost 50 lysosomal storage disorders. Ocular pathology is common in patients with MPS VI, with complications including ocular hypertension, progressive corneal clouding, optic nerve swelling (or papilledema) often associated with communicating hydrocephalus (Ashworth et al., Eye 20(5), 553-563, 2006; Goldberg et al., AJO 69(6), 969-975), and raised intracranial pressure (ICP) progressing to atrophy with loss of vision (Goodrich et al., Loss of vision in MPS VI is a consequence of increased intracranial pressure, 2002). This is the first case report of reversed papilledema and improved visual acuity in an 11-year-old MPS VI patient receiving galsulfase (Naglazyme), an enzyme replacement therapy (ERT) of recombinant human arylsulfatase B (rhASB) (Harmatz et al., J Pediatr 148(4), 533-539, 2006). PMID- 18418555 TI - Zinc or magnesium supplementation modulates cd intoxication in blood, kidney, spleen, and bone of rabbits. AB - The objective of this study was to examine the influence of oral supplementation with Zn or Mg on Cd content in the blood and organs of rabbits exposed to prolonged Cd intoxication. Rabbits were divided into the following groups: Cd group-received orally every day for 4 weeks 10 mg Cd/kg body weight (b.w.), Cd+Zn group and Cd+Mg group-exposed to Cd and supplemented with 20 mg Zn/kg b.w. or 40 mg Mg/kg b.w. 1 h after Cd treatment. Cd content in biological material was determined by atomic absorption spectrophotometry. Blood Cd concentration was determined in all investigated groups at time 0 and after 10, 14, 18, 22, 25, and 28 days, whereas Cd content in the brain, heart, lungs, liver, kidney, spleen, pancreas, skeletal muscle, and bone was determined after 28 days. Blood Cd concentration was significantly increased in all groups from the 14th day of Cd intoxication and lasted till the end of the experiment. Zn or Mg supplementation significantly reduced blood Cd content on the 18th and 25th days. Supplementation with Zn or Mg significantly decreased Cd concentration in the kidney, spleen, and bone and, in addition, Zn reduced Cd content in the brain. Supplementation with Zn or Mg in Cd-intoxicated rabbits caused similar reduction of blood Cd concentration; however, reduction of tissue Cd content was more pronounced in Zn- than in Mg-supplemented group. PMID- 18418556 TI - Erythrocyte indices, microminerals and ratios, antioxidants and lipids in Centrum Materna diet-supplemented Omani mothers. AB - Venous (maternal) and cord blood (neonatal) samples of Omani women who had a daily supplement of Centrum Materna multivitamin and multimineral tablet throughout pregnancy were investigated at late preterm (n=37) and at term (n=37) delivery for erythrocyte indices, micromineral, antioxidant, and lipid values. Hemoglobin (Hb), hematocrit (HCT), mean cell volume (MCV), red cell distribution width (RDW), copper (Cu), zinc (Zn), ceruloplasmin, erythrocyte Cu-Zn superoxide dismutase (Cu-Zn SOD), cholesterol, apolipoprotein (apo) A-I and apo B were measured by appropriate analytical systems. Cu/zinc and Cu/ceruloplasmin ratios were calculated. The erythrocyte indices were normal in neonatal blood but showed borderline anemia in maternal blood of both groups. There were significantly decreased values of Cu (P=0.012), Zn (P=0.001), apo A-I (P=0.029), and Cu/ceruloplasmin ratio (P=0.032) in late preterm compared to term mothers. Significantly decreased values of Cu (P=0.003), ceruloplasmin (P<0.0001), apo A-I (P=0.024), and Cu/Zn ratio (P=007) were observed in late preterm relative to term neonates. Late preterm mothers were significantly younger (P=0.027) than term mothers. Maternal age correlated positively with apo A-I (r=0.424, P=0.012) and negatively with Cu/Zn ratio (r=-0.353, P=0.040). The findings suggest that with daily dietary Centrum Materna supplementation throughout pregnancy, hematological indices were maintained within normal in mothers and neonates, but the levels of microminerals and micromineral ratios were subnormal in late preterm mothers and their neonates. PMID- 18418557 TI - Effects of high levels of dietary silicon on bone development of growing rats and turkeys fed semi-purified diets. AB - Two experiments were conducted using a completely randomized design to study the effects of high levels of silicon (Si) supplementation on bone development, structure, and strength in growing rats and turkeys. Rats were supplemented at two dietary Si levels: 0 and 500 ppm; and the turkeys were supplemented at four dietary Si levels: 0, 135, 270, and 540 ppm in semi-purified diets of dextrose albumin for rats and dextrose-casein for turkeys. The experiments lasted 8 and 4 weeks for the rats and turkeys, respectively. Physical, mechanical, and chemical parameters of bones were measured. All the physical and mechanical measures of bone size and strength were not different (P > 0.05) between treatments in rats and turkeys except the moment of inertia, which was lower (P < 0.01) in rats on the 500 ppm Si level of supplementation. There were small but consistent reductions in structural and strength parameters with Si supplementation which were not wholly due to differences in bodyweights of the rats and turkeys. Although bone mineral composition was not affected (P > 0.05) by Si supplementation, plasma magnesium (P = 0.08) in rats and plasma calcium (P < 0.05) in turkeys were reduced by high levels of Si supplementation. The antagonistic relations of high Si levels with calcium and magnesium were deemed to be the mechanisms through which high Si imposes its deleterious effects on bone size and strength. PMID- 18418558 TI - Large-scale analysis of protein-protein interactions using cellulose-bound peptide arrays. AB - Peptide arrays for screening large numbers of peptide fragments and probing with large numbers of samples is discussed. PMID- 18418560 TI - Multi-photon excitation imaging of dynamic processes in living cells and tissues. AB - Over the past decade, two-photon microscopy has successfully made the transition from the laser laboratory into a true biological research setting. This has been due in part to the recent development of turnkey ultrafast laser systems required for two-photon microscopy, allowing ease of use in nonspecialist laboratories. The advantages of two-photon microscopy over conventional optical sectioning techniques are for greater imaging depths and reduced overall phototoxicity, as such enabling noninvasive intra-vital imaging of cellular and subcellular processes. Greater understanding of these advantages has allowed this technique to be more effectively utilized in a biological research setting. This review will cover the recent widespread uses of two-photon microscopy and highlight the wide range of physiological studies enabled in fields such as neurosciences, developmental biology, immunology, cancer biology, and endocrinology. PMID- 18418561 TI - [Combined transgluteal ischial and femoral nerve block: retrospective data on 65 risk patients with leg amputation]. AB - For amputations of the lower limbs the combined blockade of the sciatic nerve (via the transgluteal approach) and the femoral nerve (via the inguinal route) may be a suitable alternative to general or neuroaxial anaesthetic methods. In highly comorbid, high risk patients this catheter-linked regional anaesthesia combines the advantages of avoiding controlled ventilation with decreased cardiovascular depression, as conveyed by the use of general anaesthesia or neuraxial techniques. Furthermore, improved postoperative analgesia may be achieved for several days. In our retrospective study, 65 patients who were scheduled for amputation of the lower limbs were analysed. In 58 patients (89%), a regional anaesthesia technique employing only a catheter technique was performed and only 7 patients (11%) required additional general anaesthesia. For postoperative pain management, local anaesthetics were administered via an indwelling nerve block catheter and excellent pain scores were achieved in these patients. However, successful employment of this technique necessitates sufficiently trained personnel as well as on-going training in the performance of catheter-based local analgesia. PMID- 18418563 TI - [Rectal prolapse. Abdominal or local approach]. AB - Surgical treatment of rectal prolapse aims to correct morphology and restore function. Many techniques are available, but none can be considered a gold standard. Abdominal approaches differ with regard to abdominal access, extent of rectal mobilisation, technique of rectal pexy, and concomitant sigmoid resection. Local (perineal/transanal) procedures plicate or resect the rectum. The choice of operative approach is based on the patient's condition and expected outcome of the procedure, e.g. recurrence rate, morbidity, and function. Abdominal operations are favored in fit patients, while local procedures are considered for the elderly and frail. This review compares differences in the most common techniques, focussing on recurrence, morbidity, and functional outcome. PMID- 18418564 TI - [Progress in diagnostics of anorectal disorders. Part II: radiology]. AB - Diagnostics and therapy of anorectal disorders remain a surgical question. In close cooperation between different departments (radiology and gastroenterology, urology and gynecology, dermatology and psychology), the role of radiologic imaging is of growing importance. Exact knowledge of functional anatomy and precise clinical examination constitute the basis of the according therapeutic strategies. In this context radiology has contributed decisively. Developments in imaging techniques, e.g. dynamic MRI, highly contributed to better understanding of complex functional pelvic floor disorders. The combination of nanotechnology and high-resolution imaging allows precise staging, especially in rectal cancer. Furthermore, advances in virtual colonoscopy could lead to widely acceptable and patient-friendly screening for colorectal malignancies. PMID- 18418565 TI - [Surgical therapy in Meniere's disease. Historical development and today's state of the art]. AB - After P. Meniere's first description of the typical symptoms in 1861 it took more than 40 years before the first otosurgical procedures were performed to cure Meniere's disease. Various surgical methods were established during the twentieth century, which still are employed in the treatment of intractable Meniere's disease, especially saccotomy and vestibular neurectomy but also intoxication of the labyrinth by intratympanic application of gentamicin. Despite the good results of such therapeutic regimens the basic pathological mechanism is still not fully understood. Since the description of an endolymphatic hydrops by Hallpike und Cairns in 1938 as a typical feature, there have been some observations of a possible infectious, allergic and autoimmunological (co)pathogenesis without enough proof to explain the disease in every case. This article aims to present the current scientific data, diagnostics and therapy of Meniere's disease with special emphasis on surgical treatment options. PMID- 18418566 TI - [Hormone therapy]. PMID- 18418568 TI - [Long-term results after fibroid embolization]. AB - Fibroids (leiomyoma) are the most frequent benign tumors of the uterus during female reproductive age. In the case of clinical symptoms, uterine artery embolization (UAE) is a well established treatment option for symptomatic fibroids, resulting in promising long-term results. In order to estimate long term success after UAE, reduction of fibroids and uterus volume is less important than complete improvement and disappearance of fibroid-related symptoms. In addition to a high technical success rate and a low perioperative complication rate, UAE results in high short-term and long-term patient satisfaction (>87%) and improvement of symptoms (>81%). Candidates for UAE should be informed about potential reinterventions in terms of repeated UAE (up to 18% of cases) or surgical treatment options. PMID- 18418567 TI - [Growth hormone therapy in adults. Attempt to assess a decade of use]. AB - Patients with adult growth hormone deficiency apparently can develop a clinical picture with pronounced obesity, dyslipidemia, decreased bone density, and increased fracture rate as well as psychosocial limitations irrespective of the loss and replacement of further hypophyseal axes. The extent of these changes is strongly dependent on interindividual variations. Retrospective analyses have indicated a possibility of elevated morbidity and mortality among this patient cohort which can be due to the proatherogenic alterations (central obesity, dyslipidemia). Treatment with recombinant growth hormone in controlled trials resulted in evident improvement in quality of life, better body composition and lipid profile as well as an increase in bone density. Whether these improvements will also pay off in terms of reduction of endpoints (decline in fracture rates, decrease of cardiovascular morbidity and mortality) has not yet been confirmed by controlled studies. When administered at low doses, titrated according to the IGF 1 level, growth hormone replacement appears to be a safe and well-tolerated therapeutic regimen. PMID- 18418569 TI - Elective tracheostomy in mechanically ventilated children in Canada. AB - OBJECTIVE: To determine the current practice and opinions of paediatric intensivists in Canada regarding tracheostomy in children with potentially reversible conditions which are anticipated to require prolonged mechanical ventilation. DESIGN AND SETTING: Self-administered survey among paediatric intensivists within paediatrics critical care units (PCCU) across Canada. MEASUREMENTS AND RESULTS: All 16 PCCUs participated in the survey with a response rate of 81% (63 physicians). In 14 of 16 centres one to five tracheostomies were performed during 2006. Two centres did not perform any tracheostomies. The overall rate of tracheostomy is less than 1.5%. Percutaneous technique is used in 3/16 (19%) of centres. Readiness to undertake tracheostomy during the first 21[Symbol: see text]days of illness is influenced by patient diagnosis; severe traumatic brain injury 66% vs. 42% in a 2-year-old with Guillain-Barre syndrome, 48% in a 9-year-old with Guillain-Barre syndrome, and 12% in a child with isolated ARDS. In a child with ARDS 25% of respondents would never consider tracheostomy. Age does not affect timing nor keenness for tracheostomy. The majority, 81%, believe that the risks associated with the procedure do not outweigh the potential benefits. Finally, 51% believe that tracheostomy is underutilized in children. CONCLUSIONS: Elective tracheostomy is rarely performed among ventilated children in Canada. However, 51% of physicians believe it is underutilized. The role of elective tracheostomy and the percutaneous technique in children requires further investigation. PMID- 18418570 TI - Prevention of VAP: the whole is more than the sum of its parts. PMID- 18418571 TI - Internally coated endotracheal tubes with silver sulfadiazine in polyurethane to prevent bacterial colonization: a clinical trial. AB - OBJECTIVE: Coated medical devices have been shown to reduce catheter-related infections. We coated endotracheal tubes (ETT) with silver sulfadiazine (SSD), and tested them in a clinical study to assess the feasibility, safety, and efficacy of preventing bacterial colonization. DESIGN: A prospective, randomized clinical trial, phase I-II. SETTING: Academic intensive care unit (ICU). PARTICIPANTS: Forty-six adult patients expected to need 12-24 h of intubation were randomized into two groups. INTERVENTIONS: Patients were randomized to be intubated with a standard non-coated ETT (St-ETT, n=23; control group), or with a SSD-coated ETT (SSD-ETT, n=23). MEASUREMENTS AND RESULTS: Coating with SSD prevented bacterial colonization of the ETT (frequency of colonization: SSD-ETT 0/23, St-ETT 8/23; p<0.01). No organized bacterial biofilm could be identified on the lumen of any ETT; however, SSD was associated with a thinner mucus layer (in the SSD-ETT secretion deposits ranged from 0 to 200 microm; in the St-ETT deposits ranged between 50 and 700 microm). No difference was observed between the two groups in the tracheobronchial brush samples (frequency of colonization: SSD-ETT 0/23, St-ETT 2/23; p=0.48). No adverse reactions were observed with the implementation of the novel device. CONCLUSION: SSD-ETT can be safely used in preventing bacterial colonization and narrowing of the ETT in patients intubated for up to 24 h (mean intubation time 16 h). PMID- 18418572 TI - Antimicrobial-coated endotracheal tubes: an experimental study. AB - OBJECTIVE: Antibiotic-resistant bacterial biofilm may quickly form on endotracheal tubes (ETTs) and can enter the lungs, potentially causing pneumonia. In an attempt to prevent bacterial colonization, we developed and tested in an in vitro study and animal study several antibacterial-coated ETTs (silver sulfadiazine with and without carbon in polyurethane, silver sulfadiazine and chlorhexidine with and without carbon in polyurethane, silver-platinum with and without carbon in polyurethane, chlorhexidine in polyurethane, and rose bengal for UV light). DESIGN, SETTING, ANIMALS, INTERVENTIONS: After preliminary studies, silver sulfadiazine in polyurethane (SSD-ETT) was selected among the coatings to be challenged every 24 h with 10(4)-10(6) Pseudomonas aeruginosa/ml and evaluated at 6 h, 24 h, and 72 h with standard microbiological studies, scanning electron microscopy, and confocal scanning microscopy. Subsequently, eight sheep were randomized to receive either a SSD-ETT or a standard ETT (St ETT). After 24 h of mechanical ventilation, standard microbiological studies were performed together with scanning electron microscopy and confocal microscopy. MEASUREMENTS AND RESULTS: In the in-vitro study SSD-ETT remained bacteria-free for up to 72 h, whereas St-ETT showed heavy P. aeruginosa growth and biofilm formation (p < 0.01). In sheep, the SSD-ETT group showed no bacterial growth in the ETT, ventilator tubing, and lower respiratory tract, while heavy colonization was found in the St-ETT (p < 0.01), ventilator tubing (p=0.03), and lower respiratory tract (p < 0.01). CONCLUSION: This study describes several effective and durable antibacterial coatings for ETTs. Particularly, SSD-ETT showed prevention against P. aeruginosa biofilm formation in a 72-h in-vitro study and lower respiratory tract colonization in sheep mechanically ventilated for 24 h. PMID- 18418573 TI - A "biologic technique" for the treatment of a disruption of the extensor mechanism after revision total knee arthroplasty: a case report. AB - A modified technique for the reconstruction of the extensor mechanism after revision total knee arthroplasty, is described. A medial gastrocnemius flap, in combination with in situ use of semitendinosus tendon autograft through intraosseous patellar tunnel, was used. The patient was followed for 2 years after the final reconstruction. After the procedure, the patient who previously had been dependent on a walker was able to walk without a cane. The above mentioned technique provided good functional outcome in our case. PMID- 18418574 TI - Transosseous arthroscopic repair of partial articular-surface supraspinatus tendon tears. AB - Partial articular-surface tendon avulsion (PASTA) lesions of the supraspinatus muscle represent a common cause for shoulder impairment and a preceding pathology for full-thickness tendon tears. Arthroscopic tendon repair is a possible surgical method of treatment. The purposes of cuff repair are anatomical tendon healing, prevention of tear size progression to completion and reduction of shoulder pain. In this report, we describe a transtendon arthroscopic technique of transosseous refixation of articular-side partial tears leaving the bursal layer of the supraspinatus tendon intact. A curved hollow needle is used to perform an all arthroscopic transosseous mattress suture. Thus, anatomical tendon to-bone contact of the rotator cuff to the footprint is restored. Preliminary clinical results of 16 patients are convincing with significant pain relief and functional improvement. PMID- 18418575 TI - Active non-operative treatment of acute isolated posterior cruciate ligament injury with cylinder cast immobilization. AB - The objective of our study was to investigate the outcome of an active non operative treatment of the acute isolated posterior cruciate ligament (PCL) injury using cylinder cast immobilization and brace with tibial supporter. This study included 17 acute and isolated PCL injuries with two years or more of follow-up for each patient. We applied a cylinder cast immobilization with tibial supporter with full extension of the knee for 6 weeks, and then another 6 weeks of PCL brace with tibial supporter and posterior elastic rubber band to prevent posterior sagging of the proximal tibia. There was improved anteroposterior stability with some residual laxity and improved clinical results at the last follow-up. The conservative treatment utilizing cylinder cast immobilization and brace may be one of the recommendable treatment methods in an isolated and acute PCL injury. PMID- 18418576 TI - Marked sympathetic component in the perivascular innervation of the dorsal paratendinous tissue of the patellar tendon in arthroscopically treated tendinosis patients. AB - During the recent years, a few studies have shed new light on the innervation patterns of the human patellar tendon, but the area of the loose paratendinous connective tissue dorsal to the proximal tendon proper has yet not been investigated. That is a drawback, since this is the area targeted in promising treatment regimens of chronic painful patellar tendinosis, namely sclerosing Polidocanol injection therapy, and a new surgical method conforming to ultrasound and color Doppler guided arthroscopic shaving, directed at neovessels found in the region. The present study thus aimed at investigating the paratendinous area dorsal to the proximal patellar tendon proper in seven patients being operated for tendinosis. Biopsies were collected through the new arthroscopic technique, approaching the tendon from the dorsal side. Samples were investigated using immunohistochemistry with antibodies delineating general (PGP 9.5), sensory (SP/CGRP), and sympathetic (TH/NPY) nerve patterns, and also antibodies against alpha1- and alpha2A-adrenoreceptors. Both small and large blood vessels had a marked perivascular innervation (PGP 9.5). Surprisingly, this perivascular innervation was found only to a very limited extent to correspond to sensory nerves, while there were marked immunoreactions for sympathetic markers. Adrenoreceptor immunoreactions frequently occurred in blood vessel walls. In conclusion, this study demonstrates, for the first time, the innervation patterns of the area dorsal to the patellar tendon in man. It shows that the area investigated is under marked influence by the sympathetic nervous system. Thus, sympathetic effects are likely to occur for blood vessels of the area, which is interesting since color Doppler has revealed that vessels of this area ("neovessels") display a pathologically high blood flow in tendinosis. The findings are discussed in relation to aspects of vascular regulation, and to pain symptoms of tendinosis. PMID- 18418577 TI - Polymer-hydroxyapatite composite versus polymer interference screws in anterior cruciate ligament reconstruction in a large animal model. AB - The aim of the study was to assess the hard tissue response of a composite hydroxyapatite/poly L-lactic acid (HA/PLLA) interference screw for anterior cruciate ligament (ACL) reconstruction compared to a standard PLLA screw. Twelve skeletally mature rams underwent unilateral ACL reconstruction using an autologous bone-patellar tendon graft. Each animal received either two test HA/PLLA interference screws or two control PLLA interference screws. Animals were sacrificed at 6 and 12 months post-implantation and the operated knees excised. Undecalcified sections of the screw and surrounding tissues were cut from resin embedded samples and stained; sections were approximately parallel to the longitudinal axis of the screws. A quantitative assessment of bone formation between each screw type (PLLA vs. HA/PLLA) and adjacent tissue in both the tibia and femur was undertaken using automated image analysis (KS400, Zeiss, UK). The inflammatory response of each screw type was assessed by histological evaluation. New bone formation along the perimeter of the screw threads was statistically significantly higher with the HA/PLLA than the PLLA alone. The inflammatory response as assessed semi-quantitatively by histologically determining the number of inflammatory cells present in the tissue adjacent to the implant, was higher for PLLA than HA/PLLA. Significantly increased new bone formation and decreased inflammatory cells were observed in vivo with the composite screw in comparison with the standard polymer. A novel HA/PLLA composite biomaterial in the form of an interference screw demonstrated an improved hard-tissue response compared to PLLA in a large animal ACL reconstruction. This study determined the differences in the tissue response between PLLA and a composite material of HA/PLLA. The improved tissue related outcomes observed in vivo, may be of benefit clinically in ACL reconstruction. PMID- 18418578 TI - Sternoclavicular dislocation--reconstruction with semitendinosus tendon autograft: a case report. AB - Traumatic sternoclavicular dislocation is a rare injury corresponding to less than 5% of all injuries of the scapular belt. It is preferentially treated through reduction of the sternoclavicular joint, symptom relief, a brief period of immobilization and rehabilitation, with the aim of gaining strength and range of motion. In some patients, however, this type of injury may progress with instability and pain, thus causing discomfort and pain. On such occasions, surgical treatment is chosen. The objective of this study was to report the clinical case of a sports player who progressed with chronic traumatic anterior instability of the sternoclavicular joint and underwent reconstruction using the ipsilateral semitendinosus tendon. This was a 16-year-old male patient who was a state-level judo player. Following a fall during a fight, he presented pain, slight deformity and edema in the right sternoclavicular joint, and he underwent conservative treatment for 12 months, without success. In the end, reconstruction of the sternoclavicular joint was carried out using the ipsilateral autologous semitendinosus, with resection of the intra-articular disc and suturing of the costoclavicular ligaments. We have presented a case of dislocation of the sternoclavicular joint in a high-performance judo player who underwent reconstruction using the semitendinosus, with excellent functional results after 1 year of follow-up. PMID- 18418579 TI - Cartilage repair with chondrocytes in fibrin hydrogel and MPEG polylactide scaffold: an in vivo study in goats. AB - Polylactic acid polymers have been used extensively as biomaterials and have shown promising properties for cartilage tissue engineering. Numerous scaffold materials exist and the optimal scaffold needs to be identified. We have tried to assess the possibilities for cartilage repair by the use of two different scaffold techniques; autologous chondrocytes in a fibrin hydrogel and a novel MPEG-PLGA scaffold, where autologous chondrocytes are immobilized within the MPEG PLGA scaffold by a fibrin hydrogel. Twenty adult goats were used for the study. A 6 mm circular full-thickness cartilage defect was created in both medial femoral condyles. The defects were randomized to the following four treatment groups. (1) Empty defect (control). (2) Subchondral drilling (control). (3) Fibrin hydrogel with autologous chondrocytes. (4) Fibrin hydrogel/chondrocyte solution in a MPEG PLGA porous scaffold. Animals were followed for 4 month. Eight defects in each treatment group completed the study. ICRS macroscopic scoring (0-12). Indentation test was performed to assess stiffness of repair tissue. Histological analyses was performed using O'Driscoll and Pineda cartilage scores as well as percentage tissue filling of the defects. The MPEG-PLGA/chondrocytes scaffold was the superior treatment modality based on the macroscopic surface score, histological scores and defect filling. The mechanical test demonstrated no difference between treatment groups. The MPEG-PLGA/chondrocyte composite demonstrated significantly better cartilage repair response than empty defects, osteochondral drilling and fibrin hydrogel with chondrocytes. The novel MPEG-PLGA scaffold in combination with chondrocytes need further studies with respect to longer follow-up times. PMID- 18418580 TI - Human CYP2E1 mediates the formation of glycidamide from acrylamide. AB - Regarding the cancer risk assessment of acrylamide (AA) it is of basic interest to know, as to what amount of the absorbed AA is metabolized to glycidamide (GA) in humans, compared to what has been observed in laboratory animals. GA is suspected of being the ultimate carcinogenic metabolite of AA. From experiments with CYP2E1-deficient mice it can be concluded that AA is metabolized to GA primarily by CYP2E1. We therefore examined whether CYP2E1 is involved in GA formation in non-rodent species with the focus on humans by using human CYP2E1 supersomes, marmoset and human liver microsomes and in addition, genetically engineered V79 cells expressing human CYP2E1 (V79h2E1 cells). Special emphasis was placed on the analytical detection of GA, which was performed by gas chromatography/mass spectrometry. The results show that AA is metabolized to GA in human CYP2E1 supersomes, in marmoset and human liver microsomes as well as in V79h2E1 cells. The activity of GA formation is highest in supersomes; in human liver it is somewhat higher than in marmoset liver. A monoclonal CYP2E1 human selective antibody (MAB-2E1) and diethyldithiocarbamate (DDC) were used as specific inhibitors of CYP2E1. The generation of GA could be inhibited by MAB-2E1 to about 80% in V79h2E1 cells and to about 90% in human and marmoset liver microsomes. Also DDC led to an inhibition of about 95%. In conclusion, AA is metabolized to GA by human CYP2E1. Overall, the present work describes (1) the application and refinement of a sensitive methodology in order to determine low amounts of GA, (2) the applicability of genetically modified V79 cell lines in order to investigate specific questions concerning metabolism and (3) the involvement, for the first time, of human CYP2E1 in the formation of GA from AA. Further studies will compare the activities of GA formation in genetically engineered V79 cells expressing CYP2E1 from different species. PMID- 18418581 TI - Certification and uncertainty evaluation of the certified reference materials of poly(ethylene glycol) for molecular mass fractions by using supercritical fluid chromatography. AB - Poly(ethylene glycol) (PEG) is a useful water-soluble polymer that has attracted considerable interest in medical and biological science applications as well as in polymer physics. Through the use of a well-calibrated evaporative light scattering detector coupled with high performance supercritical fluid chromatography, we are able to determine exactly not only the average mass but also all of the molecular mass fractions of PEG samples needed for certified reference materials issued by the National Metrology Institute of Japan. In addition, experimental uncertainty was determined in accordance with the Guide to the expression of uncertainty in measurement (GUM). This reference material can be used to calibrate measuring instruments, to control measurement precision, and to confirm the validity of measurement methods when determining molecular mass distributions and average molecular masses. Especially, it is suitable for calibration against both masses and intensities for matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. PMID- 18418582 TI - Sensitive HPLC-fluorescence detection of morphine labeled with DIB-Cl in rat brain and blood microdialysates and its application to the preliminarily study of the pharmacokinetic interaction between morphine and diclofenac. AB - A sensitive high-performance liquid chromatography (HPLC)-fluorescence method for determination of morphine (Mor) in rat brain and blood microdialysates was developed using 4-(4,5-diphenyl-1H-imidazol-2-yl)benzoyl chloride (DIB-Cl) as a label. Mor was labeled with DIB-Cl under mild reaction conditions (at room temperature for 10 min). The separation of DIB-Mor was carried out on an octadecylsilica (ODS) column with CH(3)CN/0.1 M acetate buffer (pH 5.4) within 14 min. The detection limits of Mor in brain and blood microdialysates at a signal to-noise ratio of 3 were 0.4 and 0.6 ng mL(-1), respectively. The proposed method was successfully applied to the preliminarily study of potential pharmacokinetic interaction between Mor and diclofenac. PMID- 18418583 TI - Analysis of microbial extracellular polysaccharides in biofilms by HPLC. Part I: Development of the analytical method using two complementary stationary phases. AB - The investigation of microbial extracellular polymeric substances (EPS) is helpful for the implementation of analytical methods which are suitable for biofilm analysis in order to understand the architecture and function of biofilms. A procedure for the qualitative and quantitative determination of various monosaccharides, oligosaccharides and uronic acids as important components of the carbohydrate fraction of microbial EPS by high-performance liquid chromatography (HPLC) and refractive index (RI)/UV detection is presented. Porous graphitic carbon and lead-form cation-exchanger have been examined as stationary phases. Therefore, two complementary HPLC methods are presented. To simulate the conditions of hydrolysis, the influences of various salts, acids and alkalis as matrix components have been investigated. Furthermore, the dependencies on the pH value and temperature of the mobile phase have been thoroughly studied. The results showed that the lead-form cation-exchanger is suitable for the separation of the neutral monosaccharides. However, for direct analysis after acidic hydrolysis with H(2)SO(4), HCl or trifluoroacetic acid, an additional purification step, e.g., precipitation or lyophilization, is necessary when the cation-exchanger is used. With the exception of hydrolysis with HCl, the porous graphitic carbon stationary phase can be used without any further purification step and is appropriate for the separation of uronic acids and their gamma-lactones. Additionally, the separation of a single monosaccharide and its derivatives is possible. Analytical parameters including the sensitivities, repeatabilities, limits of detection and limits of quantification of both HPLC methods using the RI detector are presented. The optimized method has been applied for the characterization of alginates and is also suitable for other extracellular polysaccharides in biofilms. PMID- 18418584 TI - Progress in the application of ATR-FTIR microscopy to the study of multi-layered cross-sections from works of art. AB - As a non-invasive or micro-invasive technique attenuated total reflectance Fourier transform infrared spectroscopic (ATR-FTIR) microscopy is a valuable tool for the analysis of materials in works of art. An application for which it has received growing interest is in the analysis of paint cross-sections. However, FTIR microscope configurations, objectives' geometries and low spatial resolutions, and issues of sample preparation have often hampered the characterization of individual layers or features in cross-sections. With the use of case studies, it is demonstrated here that an ATR-FTIR microscope featuring a crystal of optimized geometry and a viewing capability feature allows characterization of individual layers, or areas within layers, of 10 microm thickness or less in single measurements. Of particular value is a remote aperturing feature which allows the analysis of selected areas within the contact footprint of the ATR crystal. Since the technique is non-destructive, the same area can be analyzed by complementary microscopic techniques such as Raman spectroscopy and scanning electron microscopy with energy-dispersive spectroscopy. Pyrolysis gas chromatography-mass spectrometry was also used in some cases to corroborate the spectroscopic data. The analyses presented provided data which were important in informing art historical interpretation and conservation of the artworks examined. PMID- 18418585 TI - 12-Mercaptododecyl beta-maltoside-modified gold nanoparticles: specific ligands for concanavalin A having long flexible hydrocarbon chains. AB - A simple and highly specific protein detection system using glycoconjugated gold nanoparticles was investigated. This system was based on the aggregation of gold nanoparticles coated with carbohydrate alkanethiols in the presence of corresponding proteins (lectins) that had specific recognition for certain carbohydrates. In order to construct an efficient specific recognition system, maltoside alkanethiol was adopted as an effective sensing modifier having a disaccharide group and a flexible long alkyl chain. The surface modification of gold nanoparticles with maltoside alkanethiol resulted in a shift and broadening (from 520 to 610 nm) of the absorption peak. Monodispersed maltoside-adsorbed gold nanoparticles aggregated with the specific lectin, concanavalin A (Con A). This phenomenon was used to detect the presence of Con A and to estimate concentrations of Con A in sample solutions. The precipitate of the maltoside gold nanoparticle-Con A mixture was redispersed by addition of methyl alpha-D mannopyranoside whose adsorption coefficient is larger than that of maltoside with Con A. PMID- 18418586 TI - Characterization of interactions between polyphenolic compounds and human serum proteins by capillary electrophoresis. AB - The interaction of ten natural polyphenolic compounds (chlorogenic acid, apigenin, catechin, epicatechin, flavanone, flavone, quercetin, rutin, vicenin-2 and vitexin) with human serum albumin and mixtures of human serum albumin and alpha(1)-acid glycoprotein under near physiological conditions is studied by capillary electrophoresis-frontal analysis. Furthermore, the binding of these polyphenolic compounds to total plasmatic proteins is evaluated using ultrafiltration and capillary electrophoresis. In spite of the relatively small differences in the chemical structures of the compounds studied, large differences were observed in their binding behaviours to plasmatic proteins. The hydrophobicity, the presence/absence of some functional groups, steric hindrance and spatial arrangement seem to be key factors in the affinity of natural polyphenols towards plasmatic proteins. PMID- 18418587 TI - Studies on toxic oil syndrome: development of an enzyme-linked immunosorbent assay for 3-(N-phenylamino)propane-1,2-diol in human urine. AB - The fatty acid esters of 3-(N-phenylamino)propane-1,2-diol (PAP) are biomarkers of toxic oil batches that caused toxic oil syndrome (TOS), an intoxication that caused over 400 deaths and affected 20,000 people in Spain in 1981. PAP esters are converted into PAP by human pancreatic lipase. The in vivo biotransformation of PAP in two mouse strains generated potentially toxic metabolites. Here we report an enzyme-linked immunosorbent assay (ELISA) for PAP detection incorporating antibodies generated using PAP-hapten derivatives 1 and 2. The immunizing haptens were designed to recognize the phenylamino and hydroxymethylene moieties of the PAP structure. The antisera raised against 1-HCH showed greater affinity for free PAP, as demonstrated in competitive experiments using either 1-BSA or 2-BSA as coating antigens. The developed ELISA detects PAP at a threshold of 130 MUg L(-1) and can be used over a wide range of pH and ionic strength values. The assay can be applied to human urine samples, after a simple treatment method, with good recovery according to the correlation obtained when analyzing blind spiked urine samples. PMID- 18418588 TI - Natural wax constituents of a supercritical fluid CO(2) extract from quince (Cydonia oblonga Mill.) pomace. AB - The chemical constituents of a lipophilic extract from quince (Cydonia oblonga Mill.), obtained by supercritical fluid CO(2) extraction of the dried fruit pomace were investigated. Solvent partition of quince wax with n-hexane or acetone yielded an insoluble (crystalline) and a soluble (oily) fraction. Both fractions were analyzed separately using gas chromatography/mass spectrometry (GC/MS). The insoluble fraction consisted of saturated n-aldehydes, n-alcohols and free n-alkanoic acids of carbon chain lengths between 22 and 32, with carbon chain lengths of 26 and 28 dominating. Also odd-numbered unbranched hydrocarbons, mainly C27, C29 and C31, were detected particularly in the acetone-insoluble fraction (total, 15.8%). By means of vacuum liquid chromatography, triterpenoic acids were separated from the hexane-insoluble matter and identified as a mixture of ursolic, oleanolic and betulinic acids. The major constituents of the hexane soluble fraction were glycerides of linoleic [Delta(9,12), 18:2] and oleic [Delta(9), 18:1] acids, accompanied by free linoleic, oleic and palmitic acids (C16). Moreover beta-sitosterol, Delta(5)-avenasterol as well as trace amounts of other sterols were assigned. Finally the carotenoids phytoene and phytofluene were identified and quantified by UV/vis and high-performance liquid chromatography/MS techniques, yielding 1.0 and 0.3% of the quince wax, respectively. It is anticipated that the complex of lipid constituents from quince wax may exert interesting biological activities, the elucidation of which awaits further studies. PMID- 18418589 TI - CT demonstration of "chicken trachea" resulting from complete cartilaginous rings of the trachea in ring-sling complex. AB - We report a 10-month-old infant who presented with tetralogy of Fallot and respiratory disease in whom the suspicion of a ring-sling complex was confirmed by high-resolution CT. CT demonstrated the typical association of left pulmonary artery sling and the "chicken trachea" resulting from complete cartilaginous rings of the trachea. PMID- 18418590 TI - Chitin purification from shrimp wastes by microbial deproteination and decalcification. AB - Chitin was purified from Penaeus monodon and Crangon crangon shells using a two stage fermentation process with anaerobic deproteination followed by decalcification through homofermentative lactic acid fermentation. Deproteinating enrichment cultures from sewage sludge and ground meat (GM) were used with a proteolytic activity of 59 and 61 mg N l(-1) h(-1) with dried and 26 and 35 mg N l(-1) h(-1) with wet P. monodon shells. With 100 g wet cells of proteolytic bacteria per liter, protein removal was obtained in 42 h. An anaerobic spore forming bacterium HP1 was isolated from enrichment GM. Its proteolytic activity was 76 U ml(-1) compared to 44 U ml(-1) of the consortium. Glucose was fermented with Lactobacillus casei MRS1 to lactic acid. At a pH of 3.6, calcium carbonate of the shells was solubilised. After deproteination and decalcification of P. monodon or C. crangon shells, the protein content was 5.8% or 6.7%, and the calcium content was 0.3% or 0.4%, respectively. The viscosity of the chitin from P. monodon and C. crangon was 45 and 135 mPa s, respectively, whereas purchased crab shell chitin (practical grade) had a viscosity of 21 mPa s, indicating a higher quality of biologically purified chitin. PMID- 18418591 TI - Influence of the thyroid remnant in the elevation of the serum thyroglobulin after thyroidectomy in differentiated thyroid carcinoma. Importance of the diagnostic iodine total-body scanning. AB - PURPOSE: Stimulated thyroglobulin (Tg) and (131)I total-body scan (TBS) have been the mainstays of differentiated thyroid carcinoma (DTC), but now diagnostic TBS has been eliminated from some follow-up protocols. Nevertheless, Tg can be negative in the presence of thyroid tissue, and moderately elevated Tg poses management problems. The purpose of this study was to check how many patients have Tg negative but visible thyroid tissue and if diagnostic TBS could be clinically useful. MATERIAL AND METHODS: Retrospective review of 317 exams (stimulated Tg and TBS) of 128 patients with DTC. Patients with high-grade criteria and/or positive autoantibodies are excluded. Tg is considered positive if higher than the sensitivity level of the technique. TBS is positive if thyroid remnant, lymphatic node or metastases are detected. Results are classified in Tg and TBS concordant or discordant. RESULTS: Discordant (131)I TBS and Tg were found in 74/317 studies (23.3%), 48 (65%) being TBS positive and Tg negative. None of 128 patients had a first post-surgery negative scan, but 13 (10%) were Tg negative, even two patients with lymph node uptake. One year after radioiodine treatment, 26 out of 115 patients (initially Tg and TBS concordant positive) showed Tg negative and TBS still positive due to cervical remnants (20 patients), lymph node uptake (one) and metastasis (five); 20 patients remained Tg and TBS positive, but 14 of them only showed thyroid remnants. CONCLUSIONS: We propose to perform periodic diagnostic TBS to improve Tg specificity, at least while neck thyroid activity is detected, even if Tg is negative. PMID- 18418592 TI - Performance of integrated FDG-PET/contrast-enhanced CT in the diagnosis of recurrent ovarian cancer: comparison with integrated FDG-PET/non-contrast enhanced CT and enhanced CT. AB - PURPOSE: The aim of this study was to evaluate the accuracy of integrated positron emission tomography and computed tomography (PET/CT) using (18)F fluorodeoxyglucose with IV contrast for depiction of suspected recurrent ovarian cancer and to assess the impact of PET/contrast-enhanced CT findings on clinical management, compared with PET/non-contrast-enhanced CT and CT component. METHODS: One hundred thirty-two women previously treated for ovarian cancer underwent PET/CT consisting of non-enhanced and contrast-enhanced CT for suspected recurrence. PET/contrast enhanced CT, PET/non-contrast-enhanced CT, and enhanced CT were interpreted by two experienced radiologists by consensus for each investigation. Lesion status was determined on the basis of histopathology, radiological imaging, and clinical follow-up for longer than 6 months. RESULTS: Patient-based analysis showed that the sensitivity, specificity, and accuracy of PET/contrast-enhanced CT were 78.8% (52 of 66), 90.9% (60 of 66), and 84.8% (112 of 132), respectively, whereas those of PET/non-contrast-enhanced CT were 74.2% (49 of 66), 90.9% (60 of 66), and 82.6% (109 of 132), respectively, and those of enhanced CT were 60.6% (40 of 66), 84.8% (56 of 66), and 72.7% (96 of 132), respectively. Sensitivity, specificity, and accuracy differed significantly among the three modalities (Cochran Q test: p = 0.0001, p = 0.018, and p < 0.0001, respectively). The findings of PET/contrast-enhanced CT resulted in a change of management for 51 of the 132 patients (39%) and had an effect on patient management in 16 patients (12%) diagnosed by enhanced CT alone and three patients (2%) diagnosed by PET/non-contrast-enhanced CT. CONCLUSION: Integrated PET/contrast-enhanced CT is an accurate modality for assessing ovarian cancer recurrence and led to changes in the subsequent appropriate therapy. PMID- 18418593 TI - Individual cerebral metabolic deficits in Alzheimer's disease and amnestic mild cognitive impairment: an FDG PET study. AB - PURPOSE: The purpose of the study was the identification of group and individual subject patterns of cerebral glucose metabolism (CMRGlu) in patients with Alzheimer's disease (AD) and with amnestic mild cognitive impairment (aMCI). METHODS: [(18)F]fluorodeoxyglucose positron emission tomography (PET) studies and neuropsychological tests were performed in 16 aMCI patients (ten women, age 75+/ 8 years) and in 14 AD patients (ten women, age 75+/-9 years). Comparisons between patient subgroups and with a control population were performed using Statistical Parametric Mapping. RESULTS: Clusters of low CMRGlu were observed bilaterally in the posterior cingulate cortex (PCC), in the precuneus, in the inferior parietal lobule and middle temporal gyrus of AD patients. In aMCI patients, reduced CMRGlu was found only in PCC. Areas of low CMRGlu in PCC were wider in AD compared to aMCI and extended to the precuneus, while low CMRGlu was found in the lateral parietal cortex in AD but not in aMCI patients. Individual subject pattern analysis revealed that 86% of AD patients had low CMRGlu in the PCC (including the precuneus in 71%), 71% in the temporal cortex, 64% in the parietal cortex and 35% in the frontal cortex. Among the aMCI patients, 56% had low CMRGlu in the PCC, 44% in the temporal cortex, 18% in the frontal cortex and none in the parietal cortex. CONCLUSION: This study demonstrates that both AD and aMCI patients have highly heterogeneous metabolic impairment. This potential of individual metabolic PET imaging in patients with AD and aMCI may allow timely identification of brain damage on individual basis and possibly help planning tailored early interventions. PMID- 18418594 TI - FDG avidity and PET/CT patterns in primary gastric lymphoma. AB - PURPOSE: The use of 18F-fluoro-deoxyglucose positron emission tomography/computed tomography (FDG-PET/CT) in primary gastric lymphoma (PGL) is challenging due to physiologic FDG activity in the stomach and variability in the degree of uptake in various histologic subtypes. This study assesses FDG avidity and PET/CT patterns in newly diagnosed PGL. METHODS: Sixty-two PET/CT studies of newly diagnosed PGL were reviewed (24 low-grade mucosa-associated lymphoid tissue [MALT], 38 aggressive non-Hodgkin's lymphoma [AGNHL]). FDG avidity, patterns (focal/diffuse), and intensity (visually vs. the liver and SUVmax) were assessed and compared to 27 controls. Gastric CT abnormalities and extragastric sites were recorded. RESULTS: Gastric FDG uptake was found in 55/62 (89%) PGL (71% MALT vs. 100% AGNHL, p < 0.001) and 63% controls. A diffuse pattern was found in 60% PGL (76% MALT vs. 53% AGNHL, p = NS) and 47% controls. FDG uptake higher than liver was found in 82% PGL (58% MALT vs. 97% AGNHL, p < 0.05) and 63% controls. SUVmax in FDG-avid PGLs was 15.3 +/- 11.7 (5.4 +/- 2.9 MALT vs. 19.7 +/- 11.5 AGNHL, p < 0.001) and 4.6 +/- 1.4 in controls. CT abnormalities were found in 79% PGL (thickening, n = 49; ulcerations, n = 22). Extra-gastric FDG-avid sites were seen in none of MALT, but 61% of AGNHL (nodal, n = 18; nodal and extranodal, n = 5). CONCLUSIONS: FDG avidity was present in 89% of PGLs, including all patients with AGNHL but only 71% of MALT. FDG uptake can be differentiated, in particular in AGNHL-PGL, from physiologic tracer activity by intensity but not by pattern. Extragastric foci on PET and structural CT abnormalities are additional parameters that can improve PET/CT assessment of PGL. Defining FDG avidity and PET/CT patterns in AGNHL and a subgroup of MALT-PGL before treatment may be important for further monitoring therapy response. PMID- 18418595 TI - 18F-FDG PET/CT in sarcoidosis management: review and report of 20 cases. AB - PURPOSE: To evaluate the interest of (18)F-fluoro-2-deoxy-D: -glucose positron emission tomography/computed tomography ((18)F-FDG PET/CT) for diagnosis and therapeutic follow-up of patients with sarcoidosis. METHODS: Twenty consecutive patients with biopsy-proven sarcoidosis were retrospectively included, in particular, 13 and seven cases of thoracic and extra-thoracic sarcoidosis, respectively. All patients underwent (18)F-FDG PET/CT, and 12 of them also (67)Ga scintigraphy. Five patients were re-examined by (18)F-FDG PET/CT to assess response to corticosteroid (CS) treatment. RESULTS: Sensitivity of (18)F-FDG PET/CT in detecting active sarcoidosis localizations was determined considering only biopsy-proven sites. For thoracic, sinonasal, and pharyngo-laryngeal localizations, (18)F-FDG PET/CT sensitivity was 100%, 100%, and 80%, respectively. Overall sensitivity for all 36 biopsy-proven localizations improved from 78% to 87% after excluding skin involvement. Considering only the 12 patients who underwent both scintigraphic examinations, overall sensitivity of (67)Ga scintigraphy and (18)F-FDG PET/CT was 58% and 79%, respectively and improved to 67% and 86% after excluding all sites of skin involvement. To evaluate the efficacy of CS treatment, five enrolled patients underwent second (18)F-FDG PET/CT. Complete regression of all foci of pathological tracer uptake was showed in two cases, permitting CS withdrawal after 2 and 6 months. Improvement but incomplete regression of mediastino-pulmonary disease occurred in two patients treated with CS for 19 and 21 months. Disease progression was assessed in one patient treated with decreasing doses of CS during 16 months. CONCLUSION: (18)F-FDG PET/CT allows to obtain a complete morpho-functional cartography of inflammatory active localizations and to follow treatment efficacy in patients with sarcoidosis, particularly in atypical, complex, and multisystemic forms. PMID- 18418597 TI - Correction of oral contrast artifacts in CT-based attenuation correction of PET images using an automated segmentation algorithm. AB - PURPOSE: Oral contrast is usually administered in most X-ray computed tomography (CT) examinations of the abdomen and the pelvis as it allows more accurate identification of the bowel and facilitates the interpretation of abdominal and pelvic CT studies. However, the misclassification of contrast medium with high density bone in CT-based attenuation correction (CTAC) is known to generate artifacts in the attenuation map (mumap), thus resulting in overcorrection for attenuation of positron emission tomography (PET) images. In this study, we developed an automated algorithm for segmentation and classification of regions containing oral contrast medium to correct for artifacts in CT-attenuation corrected PET images using the segmented contrast correction (SCC) algorithm. METHODS: The proposed algorithm consists of two steps: first, high CT number object segmentation using combined region- and boundary-based segmentation and second, object classification to bone and contrast agent using a knowledge-based nonlinear fuzzy classifier. Thereafter, the CT numbers of pixels belonging to the region classified as contrast medium are substituted with their equivalent effective bone CT numbers using the SCC algorithm. The generated CT images are then down-sampled followed by Gaussian smoothing to match the resolution of PET images. A piecewise calibration curve was then used to convert CT pixel values to linear attenuation coefficients at 511 keV. RESULTS: The visual assessment of segmented regions performed by an experienced radiologist confirmed the accuracy of the segmentation and classification algorithms for delineation of contrast enhanced regions in clinical CT images. The quantitative analysis of generated mumaps of 21 clinical CT colonoscopy datasets showed an overestimation ranging between 24.4% and 37.3% in the 3D-classified regions depending on their volume and the concentration of contrast medium. Two PET/CT studies known to be problematic demonstrated the applicability of the technique in clinical setting. More importantly, correction of oral contrast artifacts improved the readability and interpretation of the PET scan and showed substantial decrease of the SUV (104.3%) after correction. CONCLUSIONS: An automated segmentation algorithm for classification of irregular shapes of regions containing contrast medium was developed for wider applicability of the SCC algorithm for correction of oral contrast artifacts during the CTAC procedure. The algorithm is being refined and further validated in clinical setting. PMID- 18418596 TI - Comparison between 68Ga-DOTA-NOC and 18F-DOPA PET for the detection of gastro entero-pancreatic and lung neuro-endocrine tumours. AB - PURPOSE: (18)F-FDG positron emission tomography (PET) value for the assessment of neuro-endocrine tumours (NET) is limited. Preliminary studies indicate that (18)F DOPA and (68)Ga-DOTA-NOC are more accurate for disease assessment and (68)Ga-DOTA peptides provide additional data on receptor status that are crucial for targeted radionuclide therapy. At present, there are no comparative studies investigating their role in NET. AIM: The aim of this study was to compare (68)Ga-DOTA-NOC and (18)F-DOPA for the evaluation of gastro-entero-pancreatic and lung neuro endocrine tumours. MATERIALS AND METHODS: Thirteen patients with biopsy-proven NET (gastro-entero-pancreatic or pulmonary) were prospectively enrolled and scheduled for (18)F-DOPA and (68)Ga-DOTA-NOC PET. PET results obtained with both tracers were compared with each other, with other conventional diagnostic procedures (CT, ultrasound) and with follow-up (clinical, imaging). RESULTS: The most common primary tumour site was the pancreas (8/13) followed by the ileum (2/13), the lung (2/13) and the duodenum (1/13). The carcinoma was well differentiated in 10/13 and poorly differentiated in 3/13 cases. (68)Ga-DOTA-NOC PET was positive, showing at least one lesion, in 13/13 cases while (18)F-DOPA PET was positive in 9/13. On a lesions basis, (68)Ga-DOTA-NOC identified more lesions than (18)F-DOPA (71 vs 45), especially at liver, lung and lymph node level. (68)Ga-DOTA-NOC correctly identified the primary site in six of eight non operated cases (in five cases, the primary was surgically removed before PET), while (18)F-DOPA identified the primary only in two of eight cases. CONCLUSIONS: Although the patients studied are few and heterogeneous, our data show that (68)Ga-DOTA-NOC is accurate for the detection of gastro-entero-pancreatic and lung neuro-endocrine tumours in either the primary or metastatic site and that it offers several advantages over (18)F-DOPA. PMID- 18418598 TI - IDO1 and IDO2 are expressed in human tumors: levo- but not dextro-1-methyl tryptophan inhibits tryptophan catabolism. AB - OBJECTIVES: Indoleamine-2,3-Dioxygenase (IDO) is an immunosuppressive molecule inducible in various cells. In addition to classic IDO (IDO1), a new variant, IDO2, has recently been described. When expressed in dendritic cells (DCs) or cancer cells, IDO was thought to suppress the immune response to tumors. A novel therapeutic approach in cancer envisages inhibition of IDO with 1-methyl tryptophan (1MT). The levo-isoform (L-1MT) blocks IDO1, whereas dextro-1MT (D 1MT), which is used in clinical trials, inhibits IDO2. Here we analyze IDO2 expression in human cancer cells and the impact of both 1-MT isoforms on IDO activity. METHODS: Surgically extirpated human primary tumors as well as human cancer cell lines were tested for IDO1 and IDO2 expression by RT-PCR. IDO1 activity of Hela cells was blocked by transfection with IDO1-specific siRNA and analysed for tryptophan degradation by RP-HPLC. The impact of D-1MT and L-1MT on IDO activity of Hela cells and protein isolates of human colon cancer were studied. RESULTS: Human primary gastric, colon and renal cell carcinomas constitutively expressed both, IDO1 and IDO2 mRNA, whereas cancer cells lines had to be induced to by Interferon-gamma (IFN-gamma). Treatment of Hela cells with IDO1-specific siRNA resulted in complete abrogation of tryptophan degradation. Only L-1MT, and not D-1MT, was able to block IDO activity in IFN-gamma-treated Hela cells as well as in protein isolates of primary human colon cancer. CONCLUSIONS: Although IDO2 is expressed in human tumors, tryptophan degradation is entirely provided by IDO1. Importantly, D-1MT does not inhibit the IDO activity of malignant cells. If ongoing clinical studies show a therapeutic effect of D-1MT, this cannot be attributed to inhibition of IDO in tumor cells. PMID- 18418599 TI - Robust hypothesis tests for independence in community assembly. AB - The extent to which competition affects the distributions of species at large spatial scales is unclear. To evaluate this question, hypothesis tests that do not depend on parametric assumptions are needed. Here, we develop a broadly applicable test that requires only one parametric assumption. Letting i and j denote the ith and jth colonists to arrive at a site, respectively, and [i j] the event that i and j belong to the same "unit" (e.g., functional group, genus), we show how colonists will be partitioned into units if for all i and j, [i j] is independent of whether i and j share unit membership with the other colonists, conditional on other information about shared units. Our distribution of partitions is useful for inferring competitive effects, because these effects predict that for at least one i and j, P ([i j]) will be less when i and j share unit membership than when they do not. PMID- 18418600 TI - The association of anti-CCP antibodies with disease activity in rheumatoid arthritis. AB - Antibodies to citrullinated proteins have been described in patients with rheumatoid arthritis (RA) and these appear to be the most specific markers of the disease. Our objective was to determine the frequency of antibodies to cyclic citrullinated peptides (CCPs) in patients with RA and the association of anti-CCP antibodies with disease activity, radiological erosions and HLA DR genotype. Forty patients with RA and 38 patients with fibromyalgia were included in this study. Serum samples were collected from both patient groups with RA and fibromyalgia. Anti-CCP was measured by the corresponding enzyme-linked immunosorbent assay. Additionally, erythrocyte sedimentation rate (ESR), C reactive protein (CRP), rheumatoid factor (RF), disease activity score (DAS), visual analog scala (VAS), HLA genotype and radiographic information were determined in patients with RA. The rate of sensitivity and specificity of anti CCP reactivity for the diagnosis RA were measured (sensitivity 50%, specificity 100%). There is no significant difference between anti-CCP (+) and anti-CCP (-) RA patients for DAS28, VAS, ESR, CRP, disease duration, HLA genotype, and radiological assessment of hand. However, there was a significant difference between anti-CCP (+) and anti-CCP (-) RA patients for RF and the radiological assessment of left and right wrists (respectively, P < 0.05, P = 0.04, P = 0.01). There was no significant correlation between anti-CCP antibody and ESR, CRP, VAS, DAS 28 or radiological assessment. A small but significant correlation was found between RF and anti-CCP antibody (P = 0.02, r = 0.35). PMID- 18418601 TI - A rabbit model of atherosclerosis at carotid artery: MRI visualization and histopathological characterization. AB - To induce a rabbit model of atherosclerosis at carotid artery, to visualize the lesion evolution with magnetic resonance imaging (MRI), and to characterize the lesion types by histopathology. Atherosclerosis at the right common carotid artery (RCCA) was induced in 23 rabbits by high-lipid diet following balloon catheter injury to the endothelium. The rabbits were examined in vivo with a 1.5 T MRI and randomly divided into three groups of 6 weeks (n=6), 12 weeks (n=8) and 15 weeks (n=9) for postmortem histopathology. The lesions on both MRI and histology were categorized according to the American Heart Association (AHA) classifications of atherosclerosis. Type I and type II of atherosclerotic changes were detected at week 6, i.e., nearly normal signal intensity (SI) of the injured RCCA wall without stenosis on MRI, but with subendothelial inflammatory infiltration and proliferation of smooth muscle cells on histopathology. At week 12, 75.0% and 62.5% of type III changes were encountered on MRI and histopathology respectively with thicker injured RCCA wall of increased SI on T(1)-weighted and proton density (PD)-weighted MRI and microscopically a higher degree of plaque formation. At week 15, carotid atherosclerosis became more advanced, i.e., type IV and type V in 55.6% and 22.2% of the lesions with MRI and 55.6% and 33.3% of the lesions with histopathology, respectively. Statistical analysis revealed a significant agreement (p<0.05) between the MRI and histological findings for lesion classification (r=0.96). A rabbit model of carotid artery atherosclerosis has been successfully induced and noninvasively visualized. The atherosclerotic plaque formation evolved from type I to type V with time, which could be monitored with 1.5-T MRI and confirmed with histomorphology. This experimental setting can be applied in preclinical research on atherosclerosis. PMID- 18418602 TI - Diffusion tensor imaging and tractography of the median nerve in carpal tunnel syndrome: preliminary results. AB - The purpose was to demonstrate the feasibility of in vivo diffusion tensor imaging (DTI) and tractography of the human median nerve with a 1.5-T MR scanner and to assess potential differences in diffusion between healthy volunteers and patients suffering from carpal tunnel syndrome. The median nerve was examined in 13 patients and 13 healthy volunteers with MR DTI and tractography using a 1.5-T MRI scanner with a dedicated wrist coil. T1-weighted images were performed for anatomical correlation. Mean fractional anisotropy (FA) and mean apparent diffusion coefficient (ADC) values were quantified in the median nerve on tractography images. In all subjects, the nerve orientation and course could be detected with tractography. Mean FA values were significantly lower in patients (p=0.03). However, no statistically significant differences were found for mean ADC values. In vivo assessment of the median nerve in the carpal tunnel using DTI with tractography on a 1.5-T MRI scanner is possible. Microstructural parameters can be easily obtained from tractography images. A significant decrease of mean FA values was found in patients suffering from chronic compression of the median nerve. Further investigations are necessary to determine if mean FA values may be correlated with the severity of nerve entrapment. PMID- 18418603 TI - The Amplatzer vascular plug for large vessel occlusion in the endovascular management of aneurysms. AB - The Amplatzer vascular plug (AVP) is derived from the Amplatzer device used in correction of cardiac septal defects. We present a large series of the use of the AVP in the endovascular management of aneurysms. Three patients with a combination of aortic and peripheral aneurysms underwent embolisation with the AVP. Plugs with 10-16 mm diameter were used and delivered using introducer/guiding sheaths. A total of 16 internal iliac arteries, 2 common iliac arteries (CIA), 4 subclavian (SCA) arteries, 1 superior mesenteric (SMA) and 1 popliteal artery were embolised. Successful occlusion with the AVP was achieved in 21 out of 24 vessels (87.5%), of which 18 (75%) occluded immediately and 3 (12.5%) were delayed occlusions. The three patients who are considered to have failed to occlude with the AVP required the use of additional embolic agents. At the end of the 17-month follow-up all 24 target vessels had occluded. Two patients developed persistent buttock claudication, and one had a left hemiparesis. The AVP is useful as an adjunctive device in the management of aneurysms. It has a particular role in embolisation of large-diameter vessels with a short implantation zone. The device is safe and easy to use. PMID- 18418604 TI - Dual-source CT coronary imaging in heart transplant recipients: image quality and optimal reconstruction interval. AB - The image quality and optimal reconstruction interval for coronary arteries in heart transplant recipients undergoing non-invasive dual-source computed tomography (DSCT) coronary angiography was evaluated. Twenty consecutive heart transplant recipients who underwent DSCT coronary angiography were included (19 male, one female; mean age 63.1 +/- 10.7 years). Data sets were reconstructed in 5% steps from 30% to 80% of the R-R interval. Two blinded independent observers assessed the image quality of each coronary segments using a five-point scale (from 0 = not evaluative to 4 = excellent quality). A total of 289 coronary segments in 20 heart transplant recipients were evaluated. Mean heart rate during the scan was 89.1 +/- 10.4 bpm. At the best reconstruction interval, diagnostic image quality (score > or = 2) was obtained in 93.4% of the coronary segments (270/289) with a mean image quality score of 3.04 +/- 0.63. Systolic reconstruction intervals provided better image quality scores than diastolic reconstruction intervals (overall mean quality scores obtained with the systolic and diastolic reconstructions 3.03 +/- 1.06 and 2.73 +/- 1.11, respectively; P < 0.001). Different systolic reconstruction intervals (35%, 40%, 45% of RR interval) did not yield to significant differences in image quality scores for the coronary segments (P = 0.74). Reconstructions obtained at the systolic phase of the cardiac cycle allowed excellent diagnostic image quality coronary angiograms in heart transplant recipients undergoing DSCT coronary angiography. PMID- 18418605 TI - Image quality on dual-source computed-tomographic coronary angiography. AB - Multi-detector CT reliably permits visualization of coronary arteries, but due to the occurrence of motion artefacts at heart rates >65 bpm caused by a temporal resolution of 165 ms, its utilisation has so far been limited to patients with a preferably low heart rate. We investigated the assessment of image quality on computed tomography of coronary arteries in a large series of patients without additional heart rate control using dual-source computed tomography (DSCT). DSCT (Siemens Somatom Definition, 83-ms temporal resolution) was performed in 165 consecutive patients (mean age 64 +/- 11.4 years) after injection of 60-80 ml of contrast. Data sets were reconstructed in 5% intervals of the cardiac cycle and evaluated by two readers in consensus concerning evaluability of the coronary arteries and presence of motion and beam-hardening artefacts using the AHA 16 segment coronary model. Mean heart rate during CT was 65 +/- 10.5 bpm; visualisation without artefacts was possible in 98.7% of 2,541 coronary segments. Only two segments were considered unevaluable due to cardiac motion; 30 segments were unassessable due to poor signal-to-noise ratio or coronary calcifications (both n = 15). Data reconstruction at 65-70% of the cardiac cycle provided for the best image quality. For heart rates >85 bpm, a systolic reconstruction at 45% revealed satisfactory results. Compared with earlier CT generations, DSCT provides for non-invasive coronary angiography with diagnostic image quality even at heart rates >65 bpm and thus may broaden the spectrum of patients that can be investigated non-invasively. PMID- 18418606 TI - Masticator space abscess derived from odontogenic infection: imaging manifestation and pathways of extension depicted by CT and MR in 30 patients. AB - Propagation of odontogenic masticator space abscesses is insufficiently understood. The purpose was to analyse pathways of spread in 30 patients with odontogenic masticator space abscess. The imaging findings in 30 patients (CT in 30, MR in 16 patients) were retrospectively analysed. CT and MR imaging depicted a masticator space abscess within: medial pterygoid muscle in 13 patients (43.3%), lateral masseter and/or pterygoid muscle in 14 (46.7%) and superficial temporal muscle in 3 patients (10%). In the lateral masticator space intra spatial abscess extension occurred in 7 of 14 patients (50%). The sub-masseteric space provided a pathway in seven (70%). Extra-spatial extension involved the submandibular space only in 3 of 14 patients (21.4%). Medial masticator space abscesses exhibited extra-spatial spread only. Extension affected the parapharyngeal space and/or soft palate in 7 of 13 lesions (53.8%). MR imaging in comparison to CT increased the number of abscess locations from 18 to 23 (27.8%) and regions affected by a cellular infiltrate from 12 to 16 (33.3%). The sub masseteric space served as a previously underestimated pathway for intra-spatial propagation of lateral masticator abscesses. Medial masticator space abscesses tend to display early extra-spatial parapharyngeal space and/or soft palate extension. PMID- 18418607 TI - Significance of sagittal reformations in routine thoracic and abdominal multislice CT studies for detecting osteoporotic fractures and other spine abnormalities. AB - The purpose was to assess osteoporotic vertebral fractures and other spinal lesions in sagittal reformations obtained from routine multidetector computed tomography (MDCT) studies of the thorax and abdomen, to compare sagittal reformations with axial images in detecting these lesions and to investigate how frequently they were missed in the official radiology report. Routine abdominal or thoracoabdominal MDCT using a standard protocol was performed in 112 postmenopausal women. Axial images and sagittal reformations were analyzed separately by two radiologists in consensus and were compared in order to evaluate how often spinal lesions could be detected. In addition the official radiology reports were assessed to determine how many of those abnormalities were identified. Spine abnormalities were visualized in 101/112 postmenopausal women. In 27 patients osteoporotic vertebral deformities were found; 6 of these were shown in the axial images, but none of these were diagnosed in the official radiology report. Additional abnormalities included degenerative disc disease, osteoarthritis of the facet joints, scoliosis, hemangiomas and bone metastases. In only 9/101 patients spine abnormalities were mentioned in the radiology report. Sagittal reformations of standard MDCT images provide important additional information on spinal abnormalities; in particular, osteoporotic vertebral deformities are substantially better detected. PMID- 18418608 TI - [Acute persistent bilateral loss of vision]. AB - Bilateral severe vision loss is a dramatic illness that requires extended diagnostics and immediate therapy. We report on a 37-year-old man who was admitted with lower back pain, headache, and fever. Within 3 days he developed meningism, pleocytosis in the cerebral spinal fluid, and bilateral vision loss, with the vision loss occurring within a few hours. Magnetic resonance imaging showed swelling of the optic nerves. No signs of infectious or immunologic disease could be detected. Despite antibiotic and antiviral treatment followed by immunosuppression with high-dose methylprednisolone, the patient's symptomatology did not decline. The disease course resulted in bilateral atrophy of the papillae caused by bilateral fulminant optic neuritis. PMID- 18418609 TI - [Surgery for acquired trochlear nerve palsy]. AB - BACKGROUND: Various surgical procedures are recommended as treatment for trochlear nerve palsy. Recession of the inferior oblique muscle (IOR), tucking or advancement of the superior oblique tendon, combination of both procedures (COP), and recession of the contralateral inferior rectus muscle are recommended. In this study, the effects of IOR and COP were compared. PATIENTS AND METHODS: Patients with isolated acquired unilateral trochlear nerve palsy were examined at a distance of 2.5 m from the Harms tangent scale before and 3 months after surgery. The onset of the palsy was 1-35 years previously (median 2 years). Subjective squint angles without diagnostic occlusion were measured with a dark red glass in front of the nonparetic eye. The field of binocular fusion was determined with an additional light bar and Bagolini striated glasses for control. The head-tilt phenomenon was defined as the difference between the vertical deviations at 45 degrees of head tilt to the right and to the left. For statistics, squint angles of left-sided palsy were transformed corresponding to palsy on the right side. RESULTS: The vertical and cyclodeviations were similar before IOR (n=13) and COP (n=21). The reduction of vertical deviation by IOR vs. COP was (median and range) 3 degrees (1; 9) vs. 6 degrees (0; 14) in primary position (PP), in side gaze 5 degrees (1; 11) vs. 9 degrees (3; 17), and in down gaze 3 degrees (-7; 11) vs. 8 degrees (2; 16). Excyclodeviation in down gaze was reduced by 4 degrees (-4; 11) vs. 7 degrees (0; 14), and the head-tilt phenomenon was improved by 1.5 degrees (-5; 7) vs. 6 degrees (-8; 14). Three months after surgery there was residual hyperdeviation of 1 degrees (0; 6) vs. 0 degrees (-7; 5) with excyclodeviation of 2 degrees (-2; 5) vs. 1 degrees (-2; 4) in PP, increasing to 2 degrees (-1; 8), ex 1 degrees (-1; 8) vs. 0 degrees (-8; 5), ex 1 degrees (-2; 5) in contralateral side gaze, and 6 degrees (-3; 13), ex 2 degrees (1; 9) vs. +1 degrees (-1; 8), ex 1 degrees (-4; 10) in down gaze. COP caused more or less significant Brown's syndrome. A second surgery was performed in one patient (4%) after COP. Augmenting surgery was done in four patients (22%) after OIR. CONCLUSIONS: Cyclovertical deviation and head-tilt phenomenon were significantly reduced when recession of the inferior oblique muscle was combined with tucking of the superior oblique tendon. To permanently minimize squint angles and abnormal head posture, initial postoperative incyclodeviation is necessary, which decreases during subsequent months due to both purely mechanical factors and modulation of cyclovertical innervation. Patient discomfort caused by this may be an argument to perform IOR as an initial procedure with fewer side effects but also fewer effects that may require further treatment. PMID- 18418610 TI - Bianchi high scrotal approach revisited. AB - Cryptorchidism is a common paediatric complaint and is traditionally managed surgically by orchidopexy. We aimed to prospectively review the success of the Bianchi high scrotal incision technique for performing orchidopexy in boys with palpable non-descended testis. A total of 206 orchidopexies were performed in 85 patients during a 5-year period. Testicular position was assessed at follow-up clinics at 6 weeks, 6 months and 2 years. Patients were also asked if they were pain free at each subsequent visit. A total of 206 Bianchi orchidopexies were performed successfully in 156 patients; one patient required an additional inguinal incision. The only post-operative complications were infections in two patients. Testes were palpable and remained in the scrotal sac after 6 weeks, 6 months and 2 years in 122 patients. Scarring was minimal and all patients and their parents were happy with the cosmetic appearance of the testes at follow-up. Our results led us to conclude that the high scrotal single incision Bianchi technique should be recommended to replace the traditional inguinal approach, which requires an additional incision in the management of undescended testis within the inguinal canal. PMID- 18418611 TI - [Periarticular mineralization of the finger]. AB - Periarticular mineralization is a clinical disorder, which is typically found at the shoulder, knee and hip joint and only rarely diagnosed at the finger joints. Periarticular ossification is a different entity and has to be distinguished from periarticular mineralization. The typical symptoms of this disorder are pain and swelling of the joint that resolves spontaneously within 3-6 months. We report on a case of periarticular mineralization of the metacarpophalangeal (MCP) joint of a 39-year-old woman. Diagnosis was made by X-ray based on findings in the form of opaque mineralizations. There was no pain relief with conservative treatment and operative treatment was performed. Histological and electron microscope analysis of the mineralization showed hydroxyapatite crystals and chondrogenic metaplasia of the surrounding fibroblasts. The patient was symptom-free soon after treatment.Usually, the therapy of the periarticular mineralization is conservative; only exceptional cases with persistent pain and swelling need operative treatment. Acute periarticular mineralization of the hand is rare and often misdiagnosed as infectious arthritis. PMID- 18418612 TI - [Rearthrodesis of the radiolunate joint]. AB - Since 1983, radiolunate arthrodesis has been the gold standard for stabilising the rheumatic wrist. Rearthrodesis of the radiolunate joint has not yet been described. In a prospective study on five radiolunate rearthrodeses with a dorsal mini titanium plate and oblique screw, bone healing was achieved in four. Fatigue fracture of the plate occurred in one case of delayed bone healing. After another rearthrodesis using the same technique, bone healing was achieved. Complete fusion of the wrist can be avoided after failed radiolunate fusion using the described operative technique for rearthrodesis of the radiolunate joint. Preserving some wrist mobility is of high value for these multimorbid patients. PMID- 18418613 TI - [Organ-specific diagnosis in patients with systemic sclerosis: Recommendations of the German Network for Systemic Sclerosis (DNSS)]. AB - The diagnosis and therapy of systemic sclerosis (SSc) is demanding due to its nature as a multisystem disease and its chronic, severe course. To date, there are no generally accepted recommendations for diagnostic work-up either for the time of initial disease diagnosis or for the regular follow-up clinical examinations and diagnostic procedures required. However, due to recent advances, e.g. in the therapy of pulmonary arterial hypertension, regular examinations may contribute to early recognition and treatment of developing organ involvement. This manuscript describes the recommendations for initial and follow-up organ specific clinical examinations and diagnostic work-up as compiled and carried out by the German Network for Systemic Scleroderma [Deutsche Netzwerk fur systemische Sklerodermie (DNSS)]. PMID- 18418614 TI - [Elevated liver enzymes in rheumatoid arthritis : differential diagnostic considerations based on a case report]. AB - Elevated liver enzymes in patients with rheumatoid arthritis may have various causes. These can range from the rheumatic disease itself, the anti-rheumatic medication or be the manifestation of an associated autoimmune disease. We present the case of a 19-year-old female with known seropositive rheumatoid arthritis who developed severe liver damage after 9 months of anti-rheumatic therapy with leflunomide and adalimumab. Both drugs were stopped. In addition to the underlying disease and the specific anti-rheumatic drugs, a temporary therapy with flucloxacillin as well as an association with newly diagnosed celiac disease had to be considered as possible causes of elevated liver enzymes. Following repeated liver biopsy, autoimmune hepatitis was assumed and prednisolone and azathioprine were initiated. Elevated liver enzymes and bilirubin rapidly returned to normal values. PMID- 18418615 TI - [Isolated tuberculosis of the tarsal bones]. AB - Although rare, isolated bone-tuberculosis of the foot merits attention. In early stages complete healing without any residual destruction can be achieved with chemotherapeutics. Therefore, when chronic inflammatory processes or tumors are biopsied, specimens should also be sent for mycobacterial testing. Tuberculosis has to be borne in mind as a cause for chronic inflammatory processes particularly in the elderly and in high risk patients (e.g. immunocompromised patients, drug abusers, HIV patients, migrants) and testing for tuberculosis has to be considered even in patients with negative tuberculin skin testing according to Mendel-Mantoux. PMID- 18418616 TI - The prevalence of absence of the palmaris longus: a study in Turkish population. AB - INTRODUCTION: It is well known that there is a wide variation in the reported prevalence of the palmaris longus (PL) absence in different ethnic groups. This prospective study was conducted to determine the prevalence of absence of PL and correlate it with gender and body side in Turkish population. METHOD: In total, 1,350 randomly selected adult patients (675 men and 675 women) who admitted to our outpatient clinic were examined for the absence of PL using Schaeffer's test and Mishra's second test. The absence of PL on both sides, results of the first and second examination tests, age, gender and dexterity were recorded and analyzed statistically. RESULTS: The overall prevalence of absence of PL (unilateral or bilateral) was 26.6% in Turkish population. The absence of PL in women was statistically more common than men. Bilateral absence of PL was statistically frequent than unilateral absence. The prevalence of absence of PL was statistically similar between the body sides. CONCLUSION: Mishra's second test which involves resisted abduction of the thumb, may be better in demonstration of the PL where the muscle was feebly developed, particularly in women. PMID- 18418617 TI - Microencapsulated human hair dermal papilla cells: a substitute for dermal papilla? AB - Dermal papillae (DP) play a pivotal role in hair formation, growth and cycling. However, the number of DP is limited. In this study, we report the production of "reconstructed DP" by enclosing DP cells within an alginate-polylysine-alginate (APA) semipermeable membrane. MTT assay and electron microscopy showed that the microencapsulated dermal papilla cells retained normal activity. The microcapsules were implanted into rat footpads, which lack follicles and sebaceous glands, to assess their inductive properties. Histologic examination showed that numbers of follicle and sebaceous gland structures formed in the footpads within 6-10-week period. At the 10 weeks following transplantation, hair fibers were visible in the footpad. These findings indicate that the DP cell microcapsules retain the capacity to initiate follicle regeneration and could be considered a substitute for fresh isolated DPs. PMID- 18418618 TI - Optimizing antenatal care and delivery in thalassemic mothers: single center experience. PMID- 18418619 TI - Effects of pyrolytic carbon on postoperative adhesion formation in rats. AB - BACKGROUND: Peritoneal adhesion is a consequence of wound healing that sometimes causes severe postoperative complications. Despite all the preventive measures and agents, adhesions have still not been eliminated completely. The aim of this study was to evaluate the effects of pyrolytic carbon on the development of postoperative peritoneal and uterine tube adhesions. MATERIAL AND METHOD: Twenty female Wistar-Albino type rats were used in this study. The rats were divided into two groups: treatment and control groups. After ketamine anaesthesia (50 mg/kg IM), a lower midline abdominal incision was performed and a standard uterine lesion was created by electrocauterization. In the study group, pyrolytic carbon was applied to the surface of the uterus and no additional procedure was applied in the control group. After 2 weeks, all of the rats were killed and uterine adhesions were staged according to the Leach scoring system. RESULTS: In the control group, the total adhesion score was 8.8 +/- 0.42. In the study group, the total adhesion score was 3.3 +/- 1.49, significantly lower than that in the control group (P < 0.001). CONCLUSION: Pyrolytic carbon significantly reduces the prevalence of peritoneal and uterine tube adhesion formation in rats. However, further more detailed investigations are needed before this material is used in clinical practice. PMID- 18418620 TI - Ectopic tubal pregnancy after hysterectomy and tubal ligation. AB - Posthysterectomy ectopic pregnancy is an unusual condition that may present soon after hysterectomy or several years later. Similarly, although tubal ligation is a widespread method of contraception, tubal pregnancy after tubal ligation is not common either. If any of these conditions are rare, having an ectopic pregnancy after hysterectomy and tubal ligation is even more infrequent and only one of such cases was found in our review of the literature. We describe the case of a 35-year-old patient, with history of bilateral tubal ligation and vaginal hysterectomy that looked for medical attention due to abdominal pain. A pregnancy test was positive and a transvaginal ultrasound demonstrated the presence of a gestational sac at the vaginal cuff, adjacent to the ovary. An exploratory laparotomy showed a ruptured ectopic pregnancy located in the distal portion of the left fallopian tube. The occurrence of an ectopic pregnancy several years following tubal ligation and vaginal hysterectomy is a rare phenomenon that appears to be secondary to a fistulous connection into the peritoneal cavity. PMID- 18418621 TI - Cystic schwannoma of the cervical plexus masquerading as a type II second branchial cleft cyst. AB - This case illustrates the rare occurrence of a cystic schwannoma of the neck, presenting clinically and ultrasonographically as a type II second branchial cleft cyst. Histology demonstrated that it was in fact a rare cystic schwannoma, most likely arising from the cervical plexus. This is the first documented case of a cystic schwannoma of the neck being mistaken for a type II second branchial cleft cyst based on clinical and ultrasound findings alone. It suggests that more sophisticated radiological investigations, such as magnetic resonance imaging, may enable accurate diagnosis of a cystic schwannoma of the neck pre-operatively. PMID- 18418622 TI - Surgery of adult bilateral vocal fold paralysis in adduction: history and trends. AB - Bilateral vocal fold paralysis (BVFP) in adduction is characterised by inspiratory dyspnea, due to the paramedian position of the vocal folds with narrowing of the airway at the glottic level. The condition is often life threatening and therefore requires surgical intervention to prevent acute asphyxiation or pulmonary consequences of chronic airway obstruction. Aside from corticosteroid administration and intubation, which are only temporary measures, the standard approach for improving respiration is to perform a tracheotomy. Over the past century, a vast majority of surgical interventions have been developed and applied to restore the patency of the airway and achieve decannulation. Surgeons can generally choose for every individual patient from various well established treatment options, which have a predictable outcome. An overview of the surgical techniques for laryngeal airway enlargement in BVFP is presented. Included are operative techniques, which have found application in clinical practice, and only to a small extent in purely anatomic or animal studies. The focus is on two major groups of interventions--for temporary and for definitive glottic enlargement. The major types of interventions include the following: (1) resection of anatomical structures; (2) retailoring and displacing the existing structures, with minimal tissue removal; (3) displacing existing structures, without tissue resection; (4) restoration or substitution of the missing innervation of the laryngeal musculature. The single interventions of these four major types have always followed the development of the medical equipment and anaesthesia. At the beginning of the twentieth century, when medicine was unable to counteract surgical infection, endoscopic or extramucosal surgical techniques were dominant. In the 1950s, the microscopic endoscopic laryngeal surgery boomed. At the end of the twentieth century many of the classical endoscopic operations were performed either with the help of surgical lasers alone, or in combination with other interventions. PMID- 18418623 TI - The radial arrangement of the human chromosome 7 in the lymphocyte cell nucleus is associated with chromosomal band gene density. AB - In the nuclei of human lymphocytes, chromosome territories are distributed according to the average gene density of each chromosome. However, chromosomes are very heterogeneous in size and base composition, and can contain both very gene-dense and very gene-poor regions. Thus, a precise analysis of chromosome organisation in the nuclei should consider also the distribution of DNA belonging to the chromosomal bands in each chromosome. To improve our understanding of the chromatin organisation, we localised chromosome 7 DNA regions, endowed with different gene densities, in the nuclei of human lymphocytes. Our results showed that this chromosome in cell nuclei is arranged radially with the gene-dense/GC richest regions exposed towards the nuclear interior and the gene-poorest/GC poorest ones located at the nuclear periphery. Moreover, we found that chromatin fibres from the 7p22.3 and the 7q22.1 bands are not confined to the territory of the bulk of this chromosome, protruding towards the inner part of the nucleus. Overall, our work demonstrates the radial arrangement of the territory of chromosome 7 in the lymphocyte nucleus and confirms that human genes occupy specific radial positions, presumably to enhance intra- and inter-chromosomal interaction among loci displaying a similar expression pattern, and/or similar replication timing. PMID- 18418624 TI - Effect of intravitreal triamcinolone acetonide on retinal apoptosis in experimental retinal neovascularization. PMID- 18418625 TI - Effort-reward imbalance, "locked in" at work, and long-term sick leave. AB - OBJECTIVES: The objective was to study the relationship between a situation characterized as being in a "locked-in" position (LIP) in occupation and/or place of work, Effort-reward imbalance (ERI), and long-term sick leave. METHODS: The study population derived from one section of a cross-sectional study SKA (sick leave, culture and attitudes), and comprised all employees at the Swedish Social Insurance Agency responsible for management and compensation of illness in the working population. The analyses were performed for 2,951 women and 534 men who had complete data. Logistic regression was used to calculate odds ratio (OR) for ERI and sick-leave, the latter only for women. RESULTS: The results showed a strong association between LIP within the place of work and ERI (for women OR = 3.28 95% CI 2.65-4.07, and for men 2.74 1.75-4.30). Also LIP within occupation resulted in high ERI (for women OR = 1.96 1.57-2.41, and for men 1.92 1.22-3.03). In women, ERI (OR = 1.40 1.15-1.70) as well as LIP within place of work (1.88 1.50-2.36) and within occupation (1.48 1.12-1.86) were associated with sick leave. ERI showed a significant mediating effect between LIP and sick leave, within place of work and within occupation (Z value 2.20 and 2.88, respectively). CONCLUSIONS: High ERI is associated with a situation characterized by being locked-in within an occupation or/and within a place of work. The results thereby support the theoretical model of Effort-reward imbalance. The results show that high ERI and being locked in are associated with long-term sick leave. ERI is a potential mediator of the association between being locked in and sick leave. PMID- 18418626 TI - Emergency spontaneous gastric perforations: ulcus versus cancer. AB - BACKGROUND: The spontaneous perforation of gastric cancer is a rare fatal complication, occurring in 1% of patients with gastric cancer, and it has a wide hospital mortality range (0-82%). In addition, it has been reported that about 10 16% of all gastric perforations are caused by gastric carcinoma. The aim of this study is to evaluate the gastric perforations and improve an alternative pathway for the management of this disorder when a pathologist is not available. MATERIAL AND METHODS: We reviewed the medical records of 513 patients who had undergone surgical treatment for gastric perforation due to gastric ulcus or gastric carcinoma in two medical centers. Sixty-seven (13.06%) patients were treated for perforated gastric carcinoma. Perforations due to trauma and iatrogenic causes were excluded. The clinicopathologic features of all patients were analyzed on the basis of their medical records. RESULTS: According to the results of our analysis, we can suggest that if a patient with gastric perforation has an age more than 60 years, an ulcus diameter (with edema) more than 6 cm, a perforation diameter more than 0.5 cm, a symptom duration of more than 20 h, and a white blood cell count less than 15.10(3)/microL, he might have a gastric carcinoma. This system has a specificity of 98.7%, a sensitivity of 53.7%, a negative predicted value of 93.4%, and positive predicted value of 85.7%. CONCLUSION: The diagnosis of malignancy is often made only on postoperative or operative frozen pathologic examination. We suggest a new pathway for the gastric perforations, if a pathologist is not available during the operation. PMID- 18418627 TI - Solitary fibrous tumour of the liver with IGF-II-related hypoglycaemia. A case report. AB - INTRODUCTION: We report a rare case of solitary fibrous tumour (SFT) of the liver associated with non-islet cell tumour hypoglycaemia (NICTH), which caused a hypoglycaemic coma due to over-production of big-insulin-like growth factor-II. DISCUSSION: Although generally benign, SFTs are better characterised, lately due to Western blot, but less than 40 cases have been reported where the liver is the target organ. In only two benign cases has hypoglycaemia been a feature. CONCLUSION: We report for the first time a demonstrable relationship between benign liver SFT and NICTH. PMID- 18418628 TI - Synovial mesenchymal stem cells accelerate early remodeling of tendon-bone healing. AB - Tendon-bone healing is important for the successful reconstruction of the anterior cruciate ligament by using the hamstring tendon. Mesenchymal stem cells (MSCs) have attracted much interest because of their self-renewing potential and multipotentiality for possible clinical use. We previously reported that MSCs derived from synovium had a higher proliferation and differentiation potential than the other MSCs that we examined. The purpose of this study was to investigate the effect and mechanism of the implantation of the synovial MSCs on tendon-bone healing in rats. Half of the Achilles' tendon grafts of rats were inserted into a bone tunnel from the tibial plateau to the tibial tuberosity with a suture-post fixation. The bone tunnel was filled with MSCs labeled with fluorescent marker DiI or without MSCs as the control. The tendon-bone interface was analyzed histologically, and collagen fibers were quantified. At 1 week, the tendon-bone interface was filled with abundant DiI-positive cells, and the proportion of collagen fiber area was significantly higher in the MSC group than in the control group. By 2 weeks, the proportion of oblique collagen fibers, which appeared to be Sharpey's fibers, was significantly higher in the MSC group than in the control group. At 4 weeks, the interface tissue disappeared, and the implanted tendon appeared to attach to the bone directly in both groups. DiI labeled cells could no longer be observed. Implantation of synovial MSCs into bone tunnel thus accelerated early remodeling of tendon-bone healing, as shown histologically. PMID- 18418629 TI - Protein expression of pigment-epithelium-derived factor in rat cochlea. AB - Pigment-epithelium-derived factor (PEDF) is a 50-kDa glycoprotein with well recognised expression in various mammalian organs showing diverse (e.g. anti angiogenic and neuroprotective) activities. However, at present, no information is available regarding the potential function of this cytokine in the inner ear. As a first approach to investigating whether PEDF is involved in cochlear function, we have explored its protein expression in the rat cochlea by immunocytochemistry. Our results show that PEDF expression in the cochlea is most prominent in the basilar membrane below the organ of Corti, in the lateral wall (especially in the stria vascularis), in ganglion neurons, and in the endothelia of blood vessels. Our findings on its distribution in the cochlea suggest that PEDF in the basilar membrane prevents blood vessel formation that would disturb cochlear micromechanics and would interfere with the mechano-electrical transduction in the organ of Corti. In cochlear ganglion neurons, PEDF might serve a neuroprotective function possibly protecting these neurons from excessive glutamate released by the inner hair cells. Our data constitute the first report on the morphological protein distribution of this multifunctional molecule in the rat cochlea and suggest its role in important functions of the internal ear. PMID- 18418630 TI - Immunohistochemical evidence for the presence of tryptophan hydroxylase in the brains of insects as revealed by sheep anti-tryptophan hydroxylase polyclonal antibody. AB - Immediately following the discovery of tryptophan hydroxylase in Drosophila, we demonstrated the presence of tryptophan hydroxylase in the brain of the beetle Harmonia axyridis (Coleoptera: Coccinellidae). However, whether tryptophan hydroxylase is present in the brains of other insects is still a matter of discussion. In the current study, sheep anti-tryptophan hydroxylase polyclonal antibody has been applied to test for tryptophan hydroxylase immunoreactivity in a broader taxonomic range of insect brains, including holometabolous and hemimetabolous insects: one species each of Coleoptera, Hymenoptera, Diptera, and Blattaria, and two species of Lepidoptera. All species show consistent tryptophan hydroxylase immunoreactivity with distribution patterns matching that of serotonin. The immuno-positive results of such an antibody in brains from diverse orders of insects suggest that specific tryptophan hydroxylase responsible for central serotonin synthesis is probably present in the brains of all insects. PMID- 18418631 TI - Regional trends for bud burst and flowering of woody plants in Norway as related to climate change. AB - Data series for bud burst, beginning of flowering and petal fall for 20 species of deciduous trees and conifers at four sites in different regions of southern Norway have been analysed and related to temperature series. On average, the spring phenophases occurred 7 days earlier during the period 1971-2005. The most significant linear trends were observed for the earliest phases. The trends in this period were compared with trends in other periods, the longest one starting in 1927. Those starting in cold decades and ending in 2005 were in most instances statistically significant, whereas hardly any significant trend appeared for series starting in warm decades. This fact showed that the results of trend studies are very sensitive to the choice of starting year. There were significant decadal variations in 40% of the series. The dates of occurrence of the phenophases, varying from the first days of May to the first days of June, correlated with seasonal temperature series, in most cases strongest to mean temperatures for the seasons March-May and April-May. The North Atlantic Oscillation Index (NAOI) for January and February appeared to have some predictive power for the date of occurrence of the recorded phases. The basis for this may be that the oscillations described by the index are of importance for the fulfilment of physiological chilling requirements needed to break bud dormancy. The same genotypes of the trees were grown in region West Norway and in Central Norwegian region; during the period 1965-2005 the trends towards earlier bud burst were more pronounced and steeper at the western site. PMID- 18418632 TI - Cellular uptake and distribution of cobalt complexes of fluorescent ligands. AB - The development of complexes that allow the monitoring of the release and distribution of fluorescent models of anticancer drugs initially bound to cobalt(III) moieties is reported. Strong quenching of fluorescence upon ligation to cobalt(III) was observed for both the carboxylate- and the hydroximate-bound fluorophores as was the partial return of fluorescence following addition of ascorbate and cysteine. The extent of the increase in the fluorescence intensity observed following addition of these potential reductants is indicative of the fluorophore being displaced from the complex by the action of ascorbate or cysteine, by ligand exchange. The cellular distribution of the fluorescence revealed that coordination to cobalt can dramatically alter the subcellular distribution of a bound fluorophore. This work shows that fluorescence can be an effective means of monitoring these agents in cells, and of determining their sites of activation. The results also reveal that the cytotoxicity of such agents correlates with their uptake and distribution patterns and that these are influenced by the types of ligands attached to the complex. PMID- 18418634 TI - Molecular insight into activated sludge producing polyhydroxyalkanoates under aerobic-anaerobic conditions. AB - One of the options enabling more economic production of polyhydroxyalkanoates compared to pure cultures is the application of mixed cultures. The use of a microbial community in a sequencing batch reactor has a few advantages: a simple process control, no necessity for sterile processing, and possibilities of using cheap substrates as a source of carbon. Nevertheless, while cultivation methods to achieve high PHAs biomass concentration and high productivity in wild and recombinant strains are defined, knowledge about the cultivation strategy for PHAs production by mixed culture and species composition of bacterial communities is still very limited. The main object of this study was to characterize on the molecular level the composition and activity of PHAs producing microorganism in activated sludge cultivated under oxygen limitation conditions. PHAs producers were detected using a PCR technique and the created PHA synthase gene library was analyzed by DNA sequencing. The obtained results indicate that PHAs-producers belonged to Pseudomonas sp., and possessed genes coding for mcl-PHA synthase. The kinetics of mcl-PHA synthase expression was relatively estimated using real-time PCR technology at several timepoints. Performed quantitative and qualitative analysis of total bacterial activity showed that there were differences in total activity during the process but differential expression of various groups of microorganisms examined by using DGGE was not observed. PMID- 18418633 TI - Bis-histidine-coordinated hemes in four-helix bundles: how the geometry of the bundle controls the axial imidazole plane orientations in transmembrane cytochromes of mitochondrial complexes II and III and related proteins. AB - Early investigation of the electron paramagnetic resonance spectra of bis histidine-coordinated membrane-bound ferriheme proteins led to the description of a spectral signal that had only one resolved feature. These became known as "highly anisotropic low-spin" or "large g(max)" ferriheme centers. Extensive work with small-molecule model heme complexes showed that this spectroscopic signature occurs in bis-imidazole ferrihemes in which the planes of the imidazole ligands are nearly perpendicular, deltaphi = 57-90 degrees. In the last decade protein crystallographic studies have revealed the atomic structures of a number of examples of bis-histidine heme proteins. A frequent characteristic of these large g(max) ferrihemes in membrane-bound proteins is the occurrence of the heme within a four-helix bundle with a left-handed twist. The histidine ligands occur at the same level on two diametrically opposed helices of the bundle. These ligands have the same side-chain conformation and ligate heme iron on the bundle axis, resulting in a quasi-twofold symmetric structure. The two non-ligand-bearing helices also obey this symmetry, and have a conserved small residue, usually glycine, where the edge of the heme ring makes contact with the helix backbones. In many cases this small residue is preceded by a threonine or serine residue whose side-chain hydroxyl oxygen acts as a hydrogen-bond acceptor from the N(delta1) atom of the heme-ligating histidine. The deltaphi angle is thus determined by the common histidine side-chain conformation and the crossing angle of the ligand-bearing helices, in some cases constrained by hydrogen bonds to the serine/threonine residues on the non-ligand-bearing helices. PMID- 18418635 TI - Delta band contribution in cue based single trial classification of real and imaginary wrist movements. AB - The aim of this study was to classify different movements about the right wrist. Four different movements were performed: extension, flexion, pronation and supination. Two-class single trial classification was performed on six possible combinations of two movements (extension-flexion, extension-supination, extension pronation, flexion-supination, flexion-pronation, pronation-supination). Both real and imaginary movements were analysed. The analysis was done in the joint time-frequency domain using the Gabor transform. Feature selection was based on the Davis-Bouldin Index (DBI) and feature classification was based on Elman's recurrent neural networks (ENN). The best classification results, near 80% true positive rate, for imaginary movements were achieved for discrimination between extension and any other type of movement. The experiments were run with 10 able bodied subjects. For some subjects, real movement classification rates higher than 80% were achieved for any combination of movements, though not simultaneously for all six combinations of movements. For classification of the imaginary movements, the results suggest that the type of movement and frequency band play an important role. Unexpectedly, the delta band was found to carry significant class-related information. PMID- 18418638 TI - Bone in celiac disease. AB - Chronic inflammation and malabsorption in celiac disease (CD) can cause bone metabolism alterations and bone mineral loss in children and adults. Bone status before and after gluten-free diet, epidemiology of fractures, and possible treatment options for CD-related osteoporosis are presented. Controversial aspects of this complication of CD are discussed. The relationship between bone derangements and celiac disease (CD) was recognized almost 50 years ago, but many questions are still open. We are now aware that osteoporosis is a relatively frequent atypical presentation of CD, especially in adults, and that undiagnosed CD can be the cause of osteoporosis and related fractures. Chronic inflammatory intestinal diseases, including CD, can affect bone and mineral metabolism because of alterations in both systemic and local regulatory factors. The pathogenetic processes are still controversial, but two main mechanisms seem to be involved: intestinal malabsorption and the presence of chronic inflammation. This review analyzes the published data on bone involvement in children, adolescents, and adults either before or after a gluten-free diet. Special attention is paid to the epidemiology of fractures in celiac patients, considering that fractures are a major complication of osteoporosis and an important problem in the management of a chronic disease like CD. The usefulness of screening osteoporotic patients systematically for CD is still an open question, but some rules can be given. Finally, the current treatment options for children and adults are discussed. Recommendations for future clinical research are proposed. PMID- 18418639 TI - Replication of associations between LRP5 and ESRRA variants and bone density in premenopausal women. AB - Replication is a critical step to validate positive genetic associations. In this study, we tested two previously reported positive associations. The low density lipoprotein receptor-related protein 5 (LRP5) Val667Met and lumbar spine bone density are replicated. This result is in line with results from large consortiums such as Genomos. However, the estrogen-related receptor alpha (ESRRA) repeat in the promoter is not replicated although the polymorphism studied was functional and could have been a causative variant. INTRODUCTION: We sought to validate associations previously reported between LRP5 V667M polymorphism and lumbar spine (LS, p = 0.013) and femoral neck (FN, p = 0.0002) bone mineral density (BMD), and between ESRRA 23 base pair repeat polymorphism and LS BMD (p = 0.0036) in a sample of premenopausal Caucasian women using an independent sample. METHODS: For the replication sample, we recruited 673 premenopausal women from the Toronto metropolitan area. All women were Caucasian and had BMD measured. LRP5 V667M was genotyped by allele-specific PCR and ESRRA repeats by sizing of PCR products on agarose gels. RESULTS: We reproduced the same association as we reported previously between LRP5 V667M and LS BMD (p = 0.015) but not with FN BMD (p = 0.254). The combined data from the two populations indicate an effect size of 0.28SD for LS BMD (p = 0.00048) and an effect size of 0.26 SD for FN BMD (p = 0.00037). In contrast, the association we reported earlier between ESRRA repeats and LS BMD was not replicated in the sample from Toronto (p = 0.645). CONCLUSIONS: The association between LRP5 V667M and LS BMD is confirmed but not that between ESRRA repeats and LS BMD. This result indicates that it is imperative to validate any positive association in an independent sample. PMID- 18418640 TI - Interactions of interleukin-6 gene polymorphisms with calcium intake and physical activity on bone mass in pre-menarche Chinese girls. AB - This study assessed independent associations and interactions of IL-6 promoter alleles (-174G/C and -634C/G), calcium intake and physical activity with bone mass among pre-menarche Chinese girls. The -634 CC carriers, greater calcium intake and physical activity were associated with better bone mass. The gene-bone association was more pronounced among girls with high physical activity or with low calcium intake. INTRODUCTION: The association between interleukin (IL)-6 promoter polymorphisms and bone mass remains in debate. This cross-sectional study examined the association between the IL-6 promoter alleles (-174G/C and 634C/G) and bone mass, and assessed if the association could be modified by calcium intake or physical activity in pre-menarche Chinese girls. METHODS: Two hundred and twenty-eight healthy pre-menarche girls aged 9-11 years were recruited from primary schools in Guangzhou, China by sending letters to parents. None of them had diseases or medications known to affect bone metabolism. The IL 6 promoter genotypes were determined by PCR-RFLP, and BMD and BMC at the total body, lumbar spine, total hip and femoral neck were measured by DXA. Calcium intake and physical activity were assessed by face-to-face questionnaire interview. RESULTS: One hundred and seventy-six subjects completed the entire study. We did not detect gene polymorphism at the IL-6 -174G/C locus, all were GG homozygotes. The IL-6 -634C/G polymorphism was significantly associated with both BMD and BMC even after adjusting for age and weight. Girls with CC genotype had higher levels of BMC and BMD than G allele carriers (+8.3% for the total body BMC, and +2.9%, +5.8%, and +5.7% for BMDs at the total body, total hip, and femoral neck, respectively; P < 0.05). The favorable effect of physical activity on BMDs at the total hip and femoral neck was much more pronounced in CC carriers than in G allele carriers, and the CC genotype associated higher BMDs at the total hip and femoral neck were observed only in girls with high level physical activity (P for interactions = 0.036 and 0.021, adjusted for age and weight). Calcium had a more benefit to the total body BMC in G allele carriers than in CC carriers, and the G allele-associated lower total body BMC was found only in subjects with low calcium intake. CONCLUSION: The IL-6 -634C/G polymorphism was significantly associated with BMD and the association might be modified by calcium intake or physical activity in pre-menarche Chinese girls. PMID- 18418641 TI - Adverse effects reported in pulsed dye laser treatment for port wine stains. AB - A retrospective study was conducted over a 762-day period to investigate the incidence of adverse effects following pulsed dye laser treatment for port wine stains using the Chromos 585 nm pulsed dye laser (PDL). Treatment energy densities typically ranged from 4.0 J/cm(2) to 7.5 J/cm(2). Treatment was predominantly delivered as a 7 mm-diameter pulse of laser light, although a 5 mm pulse was occasionally used. Nine patients experienced one adverse event each, with scabbing, blistering (prolonged in one case that persisted for more than 2 months), hypopigmentation or hyperpigmentation; one of the nine patients developed both blistering and hypopigmentation. The incidence of adverse effects per treatment was 1.4% (1.7% for adults and 0.6% for children). The lower leg was the most common site for an adverse effect; four out of 22 patients (18%) receiving PDL treatment for lower leg port wine stains (PWSs) suffered an adverse effect (three cases of hyperpigmentation and one of hypopigmentation and blistering). Four patients suffered an adverse effect following an increase in the energy density of the laser treatment. There was no statistically significant correlation between laser performance and the occurrence of adverse effects. PMID- 18418642 TI - Symptoms treatment for allergic rhinitis using diode laser: results after 6-year follow-up. AB - Allergic rhinitis is a very common chronic disease. The purpose of this study was to measure the outcome of treating allergic rhinitis with diode laser, over a long-term follow-up of 6 years. Forty-two consecutive patients with newly diagnosed allergic rhinitis that was refractive to conservative medical therapies were enrolled in the study. Treatment was with a diode laser (wavelength 805 nm, laser parameter 12 W, in 'contact' mode) under local anesthesia between September 1999 and May 2000. A quality of life questionnaire was used after a follow-up period of 6 years to assess the outcome of diode laser surgery. All 42 patients suffered from either perennial or persistent-type allergic rhinitis. Twenty-five out of 42 patients (60%) replied to the questionnaire 6 years after laser surgery. The mean operation time was 1 min 28 s for each patient. No complications were observed (e.g., major bleeding), and no nasal packing was necessary. Statistical analysis revealed significant improvements in symptom score, medication score and total score 6 years after laser surgery. A total of 60% of the patients described improvements in symptom scores, and 68% of the patients described improvements in total score 6 years after laser treatment. It was evident that the effectiveness of laser surgery persisted through the 6 years of follow-up. Long-term follow-up after 6 years confirmed that diode laser surgery for allergic rhinitis is a useful procedure that has a long-lasting effect and can be performed in an outpatient surgery under local anesthesia. Minimal discomfort, long-term effectiveness and short operation time are other reasons why this treatment should be selected for patients with allergic rhinitis that is refractive to conservative management. PMID- 18418643 TI - Bone repair following bone grafting hydroxyapatite guided bone regeneration and infra-red laser photobiomodulation: a histological study in a rodent model. AB - The aim of the investigation was to assess histologically the effect of laser photobiomodulation (LPBM) on a repair of defects surgically created in the femurs of rats. Forty-five Wistar rats were divided into four groups: group I (control); group II (LPBM); group III (hydroxyapatite guided bone regeneration; HA GBR); group IV (HA GBR LPBM). The animals in the irradiated groups were subjected to the first irradiation immediately after surgery, and it was repeated every day for 2 weeks. The animals were killed 15 days, 21 days and 30 days after surgery. When the groups irradiated with implant and membrane were compared, it was observed that the repair of the defects submitted to LPBM was also processed faster, starting from the 15th day. At the 30th day, the level of repair of the defects was similar in the irradiated groups and those not irradiated. New bone formation was seen inside the cavity, probably by the osteoconduction of the implant, and, in the irradiated groups, this new bone formation was incremental. The present preliminary data seem to suggest that LPMB therapy might have a positive effect upon early wound healing of bone defects treated with a combination of HA and GBR. PMID- 18418644 TI - Dietary toxicity and tissue accumulation of methylmercury in American kestrels. AB - American kestrels (Falco sparverius) were fed meat diets containing 0, 3, 6, or 12 ppm (dry weight) methylmercury chloride. Birds fed the 12-ppm diet started to show signs of neurotoxicity after 26 days and all died in 39-49 days. One male kestrel fed the 6-ppm diet died after 75 days of exposure and several others showed signs of neurotoxicity after 45 days. None of the birds fed the 3-ppm diet died or showed signs of toxicity. After 59 days of exposure, mercury concentrations in the liver, kidney, and blood of nonreproducing kestrels increased with increasing dietary concentration. Tissue concentrations of mercury also steadily increased over time in birds fed diets with 6 ppm mercury, which were necropsied at 8, 15, 29, or 59 days of exposure, reaching mean total mercury concentrations of 57, 46, and 45 ppm (wet weight) at 59 days in the liver, kidney, and whole blood, respectively. Two pairs of kestrels at each dietary concentration were allowed to breed. Eggs averaged 8.3 and 18.1 ppm (wet weight) total mercury from birds fed 3- and 6-ppm diets, respectively. Feathers grown during mercury exposure contained high concentrations of mercury: Birds fed 3- and 6-ppm diets contained 275 and 542 ppm total mercury, respectively. PMID- 18418645 TI - Differential pulmonary perfusion scan after percutaneous occlusion of the patent ductus arteriosus: one-decade consecutive longitudinal study from a single institution. AB - Reduced left lung perfusion has been described following percutaneous occlusion of the patent ductus arteriosus (PDA). We aimed to identify the incidence of lung perfusion abnormalities and the associated risk factors in our consecutive series. Between November 1994 and December 2003, 150 procedures were performed on 145 patients, age 4.6 +/- 4 years. Gianturco coil was used in 88.2%, Amplatzer duct occluder in 6.7%, and Rashkind Umbrella in 5.5%. Lung perfusion scan was scheduled within 48 h (LPS-1), at 6-12 months (LPS-2) and later (LPS-3) in the case of persistent abnormalities. Left lung perfusion <40% was considered abnormal. LPS-1 was obtained in 95.8% and was abnormal in 31%. LPS-2, available in 48.2%, returned to normal in 65.7% (p < 0.001). LPS-3, required in 6.2%, was normal in 55.6% (p = 0.07). Identifiable risk factors were low age and height (p < 0.01), higher Q(p)/Q(s) ratio (p < 0.05), and larger PDA size indexed for height (p < 0.001) or body surface area (p < 0.01). The number of coils or loops deployed in the pulmonary end of the PDA did not influence lung perfusion. In conclusion, we describe a high incidence of left lung perfusion reduction following percutaneous PDA occlusion, more likely in the young with large PDA. However, spontaneous recovery usually occurs within a few months. PMID- 18418647 TI - Intralesional triamcinolone acetonide for keloid treatment: a systematic review. PMID- 18418648 TI - The efficacy of melilotus extract in the management of postoperative ecchymosis and edema after simultaneous rhinoplasty and blepharoplasty. AB - BACKGROUND: Melilotus extract has been used as a folk medicine in inflammation related therapy. This study was undertaken to determine the potential benefits of this herbal extract in treating postoperative ecchymosis and edema after simultaneous rhinoplasty and blepharoplasty. METHODS: Augmentation rhinoplasty and double-eyelid blepharoplasty were performed in one procedure by the same surgeon on 46 patients. RESULTS: Postoperative edema and ecchymosis were evaluated independently by two investigators checking the upper eyelid, lower eyelid, and paranasal area at postoperative days 1, 4, and 7. Our data showed that the patients receiving melilotus extract (n = 16) had lower scores not only in upper-eyelid and lower-eyelid ecchymosis, but also in paranasal ecchymosis at postoperative day 7 when compared with those in the control group. No significant difference in postoperative ecchymosis was detected in the group receiving dexamethasone treatment (n = 16) when compared with the control group (n = 14) at postoperative days 1, 4, and 7. The mean scores of upper-eyelid, lower-eyelid, and paranasal edema at postoperative days 1 and 4 were significantly lower in the group receiving dexamethasone treatment (n = 16) than that in the control group. Although the scores of eyelid edema reduction in the patients receiving melilotus extract showed no difference when compared with the control group, paranasal edema was reduced much more at postoperative days 4 and 7 when compared with the control group. CONCLUSION: We suggest the appropriate use of melilotus extract based on its beneficial effects on the postoperative results after simultaneous rhinoplasty and blepharoplasty. PMID- 18418649 TI - Fluoroscopy-guided removal of pull-type gastrostomy tubes. AB - These case reports demonstrate a radiologic interventional technique for removal of pull-type gastrostomy tubes. This approach proved to be a safe and efficient procedure in two patients. The procedure may be applicable in situations where endoscopic attempts fail. PMID- 18418650 TI - Actinomycetal community structures in seawater and freshwater examined by DGGE analysis of 16S rRNA gene fragments. AB - The actinomycetal community structures in marine and freshwater environments (the Pacific Ocean, East China Sea, Tokyo Bay, and Arakawa River) were investigated by a culture-independent molecular method to clarify spatial and seasonal distributions. Deoxyribonucleic acid (DNA) was extracted from environmental water samples, and a community analysis was carried out on polymerase chain reaction amplified 16S ribosomal DNA. The amplified DNA fragments were analyzed by denaturing gradient gel electrophoresis (DGGE) and nonmetric multidimensional scaling analysis, followed by sequencing analysis. The actinomycetal community structures were different at each station in the Pacific Ocean, the East China Sea, Tokyo Bay, and Arakawa River, and different populations predominated in each area. There were vertical variations in actinomycetal communities in the Pacific Ocean and East China Sea between the surface and 100-m depth, but communities were similar from 200- to 1,000-m depths. There were also distinct seasonal variations in communities in Tokyo Bay. Phylogenetic analysis of DNA fragments recovered from DGGE bands revealed that most of the predominant actinomycetal strains were uncultured and were quite different from well known culturable strains, such as the Streptomyces, Micromonospora, Microbispora, Salinispora, and Actinoplanes groups. These results suggest that the marine environment is an attractive target for discovering new actinomycetal populations producing bioactive compounds and that sampling depth and season are important considerations for isolating various populations effectively. PMID- 18418651 TI - The effect of a hyaluronan-carboxymethylcellulose membrane vs. polyglactin 910 mesh on intra-abdominal tumor formation in mice. AB - PURPOSE: Hyaluronan mediates growth of SW620 colon cancer cells. Because hyaluronan is the active ingredient in Seprafilm, we hypothesized that Seprafilm would affect intraperitoneal tumor growth in a mouse model of peritoneal seeding. METHODS: Immunodeficient mice underwent laparotomy and intraperitoneal inoculation of 10(5) SW620 cells. Seprafilm (n = 22), Vicryl mesh (foreign body control; n = 24), or no material (sham; n = 19) was placed under the incision. Mice were killed after four weeks and tumors were dissected, counted, and weighed. RESULTS: Ninety-five percent of mice in the sham group and 96 percent in the Vicryl group developed intraperitoneal tumors. In contrast, only 64 percent of mice in the Seprafilm group developed tumors (P = 0.024), and these tumors were smaller than those in the sham group; (Seprafilm = 42 +/- 9 mg vs. sham = 82 +/- 17 mg; P = 0.05). In contrast, tumors in the Vicryl group were dramatically larger (349 +/- 49 mg; P < 0.001 vs. sham or Seprafilm). CONCLUSIONS: Despite previous data that suggested that hyaluronan increases colon cancer cell growth, we found that Seprafilm decreased tumor formation and tended to decrease size in this model. In contrast, Vicryl mesh increased tumor formation and size. Our results suggest that Seprafilm does not promote intraperitoneal tumor growth, especially compared with Vicryl mesh. PMID- 18418652 TI - Fistula-associated anal adenocarcinoma: good results with aggressive therapy. AB - PURPOSE: To evaluate the clinical features, pathology, treatment, and outcome of patients with fistula-associated anal adenocarcinoma. METHODS: We identified 14 patients with histologically proven fistula-associated anal adenocarcinoma. We reviewed their medical records and pathology specimens to characterize their presentation, treatment, and clinical outcome. RESULTS: Nine patients presented with a persistent fistula, 3 with a perianal mass, 1 with pain and drainage, and 1 with a recurrent perianal abscess. The average age at time of diagnosis was 59 (range, 37-76) years. Eleven patients had preexisting chronic anal fistulas. Ten had Crohn's disease, and 1 had previously received pelvic radiation therapy. The diagnosis of cancer was suspected during physical examination in 6 of the 14 patients (43 percent). Twelve patients had extensive local disease at presentation. Primary abdominoperineal resection was performed in 11 patients, 7 following neoadjuvant chemoradiation. Six patients received postoperative chemotherapy, and 2 received postoperative radiation. Four patients died with metastatic disease. The remaining 10 patients are alive without evidence of disease at a mean follow-up of 64.3 (range, 14-149) months. CONCLUSIONS: The diagnosis of fistula-associated anal adenocarcinoma is often unsuspected. Most patients can be cured with aggressive surgical and adjuvant chemoradiotherapy. PMID- 18418653 TI - Hand-assisted laparoscopic vs. laparoscopic colorectal surgery: a multicenter, prospective, randomized trial. AB - PURPOSE: This study was designed to compare short-term outcomes after hand assisted laparoscopic vs. straight laparoscopic colorectal surgery. METHODS: Eleven surgeons at five centers participated in a prospective, randomized trial of patients undergoing elective laparoscopic sigmoid/left colectomy and total colectomy. The study was powered to detect a 30-minute reduction in operative time between hand-assisted laparoscopic and straight laparoscopic groups. RESULTS: There were 47 hand-assisted patients (33 sigmoid/left colectomy, 14 total colectomy) and 48 straight laparoscopic patients (33 sigmoid/left colectomy, 15 total colectomy). There were no differences in the patient age, sex, body mass index, previous surgery, diagnosis, and procedures performed between the hand-assisted and straight laparoscopic groups. Resident participation in the procedures was similar for all groups. The mean operative time (in minutes) was significantly less in the hand-assisted laparoscopic group for both the sigmoid colectomy (175 +/- 58 vs. 208 +/- 55; P = 0.021) and total colectomy groups (time to colectomy completion, 127 +/- 31 vs. 184 +/- 72; P = 0.015). There were no apparent differences in the time to return of bowel function, tolerance of diet, length of stay, postoperative pain scores, or narcotic usage between the hand-assisted laparoscopic and straight laparoscopic groups. There was one (2 percent) conversion in the hand-assisted laparoscopic group and six (12.5 percent) in the straight laparoscopic group (P = 0.11). Complications were similar in both groups (hand-assisted, 21 percent vs. straight laparoscopic, 19 percent; P = 0.68). CONCLUSIONS: In this prospective, randomized study, hand-assisted laparoscopic colorectal surgery resulted in significantly shorter operative times while maintaining similar clinical outcomes as straight laparoscopic techniques for patients undergoing left-sided colectomy and total abdominal colectomy. PMID- 18418654 TI - Prognosis for proctology. PMID- 18418655 TI - Temporary sacral nerve stimulation for treatment of irritable bowel syndrome: a pilot study. AB - PURPOSE: This study was designed to evaluate the effect of temporary sacral nerve stimulation in patients with diarrhea-predominant irritable bowel syndrome. METHODS: Symptoms of diarrhea-predominant irritable bowel syndrome and disease specific quality of life was evaluated in six patients before and during percutaneous sacral nerve evaluation test. Primary end points were differences between total irritable bowel syndrome symptom score and total quality of life score before and during stimulation. Secondary end points were differences between the variable domains. RESULTS: Percutaneous sacral nerve evaluation test was performed in five women and one man (median age, 33 (range, 26-54) years). The irritable bowel syndrome symptom score decreased from 48.9 to 28.3 (P = 0.004). Pain, bloating, and diarrhea were significantly reduced from 7.9, 13.5, and 17.3 to 4.4, 7.2, and 10.6, respectively (P = 0.02, P = 0.01, P = 0.03). The irritable bowel syndrome quality of life score decreased from 99.3 to 59.6 (P = 0.009). Daily activities, emotional distress, eating habits, and fatigue were significantly reduced from 26.9, 22.2, 15.2, and 23.2 to 16.9, 13.3, 8, and 14.4, respectively (P = 0.02, P = 0.02, P = 0.02, P = 0.007). Two weeks after cessation of stimulation, the patients had symptoms as before stimulation. CONCLUSIONS: Temporary sacral nerve stimulation provides a significant reduction in diarrhea predominant irritable bowel symptoms and improves quality of life. Further studies with permanent implantation and double-blind crossover ON-and-OFF stimulation to evaluate the impact of placebo effect are needed. PMID- 18418656 TI - Diabetes mellitus affects response to neoadjuvant chemoradiotherapy in the management of rectal cancer. AB - INTRODUCTION: Although diabetic patients with rectal cancer have poorer outcomes than their nondiabetic counterparts, few studies have looked at diabetics' response to therapy as an explanation for this disparity. This study compares the neoadjuvant chemoradiotherapy (CRT) response in diabetic and nondiabetic patients with locally advanced rectal cancers. METHODS: This is a single-institution, retrospective review of rectal cancer patients who received CRT followed by resection from 1995 to 2006. Pretreatment tumor-node-metastasis (TNM) staging was determined using endorectal ultrasound, computed tomography (CT) scan, and magnetic resonance imaging (MRI); post-treatment staging was determined by pathological review. RESULTS: 110 patients were included; seventeen had diabetes and 93 were nondiabetics. Pretreatment staging was similar in both groups. Sixteen of the diabetics (94%) completed CRT compared to 92% (86/93) of the nondiabetics. Tumor downstaging rates were similar in the two groups (53% in diabetics, 52% in nondiabetics). Nondiabetic patients had a higher rate of nodal downstaging although not statistically significant (67% versus 27%, P = 0.80). While none of the diabetics patients achieved a pathologic complete response (pCR), 23% (21/93) of the nondiabetics did (P = 0.039). Local progression rates were higher in the diabetic group (24% versus 5%, P = 0.046). CONCLUSION: Our study shows that neoadjuvant chemoradiotherapy in rectal cancer is less effective in diabetic patients than in nondiabetics. While minimal differences are found in the rate of downstaging, the rate of achieving a complete pathologic response was significantly higher in nondiabetic patients, and in fact was not seen in any of our diabetic patients. This may explain the poorer outcomes seen in diabetic patients with rectal cancer. PMID- 18418657 TI - Transient responses to spatial perturbations in advective systems. AB - We study the transient dynamics, following a spatially-extended perturbation of models describing populations residing in advective media such as streams and rivers. Our analyses emphasize metrics that are independent of initial perturbations-resilience, reactivity, and the amplification envelope-and relate them to component spatial wavelengths of the perturbation using spatial Fourier transforms of the state variables. This approach offers a powerful way of understanding the influence of spatial scale on the initial dynamics of a population following a spatially variable environmental perturbation, an important property in determining the ecological implications of transient dynamics in advective systems. We find that asymptotically stable systems may exhibit transient amplification of perturbations (i.e., have positive reactivity) for some spatial wavelengths and not others. Furthermore, the degree and duration of amplification varies strongly with spatial wavelength. For two single population models, there is a relationship between transient dynamics and the response length that characterizes the steady state response to spatial perturbations: a long response length implies that peak amplification of perturbations is small and occurs fast. This relationship holds less generally in a specialist consumer-resource model, likely due to the model's tendency for flow induced instabilities at an alternative characteristic spatial scale. PMID- 18418658 TI - The revolution will be televised. PMID- 18418659 TI - Gastroesophageal reflux in patients with morbid obesity: a role of obstructive sleep apnea syndrome? AB - BACKGROUND: Obesity is a risk factor for gastroesophageal reflux disease (GERD) and for obstructive sleep apnea (OSA). Our aim was to evaluate in morbidly obese patients the prevalence of OSA and GERD and their possible relationship. METHODS: Morbidly obese patients [body mass index (BMI) >40 or >35 kg/m(2) in association with comorbidities] selected for bariatric surgery were prospectively included. Every patient underwent a 24-h pH monitoring, esophageal manometry, and nocturnal polysomnographic recording. RESULTS: Sixty-eight patients [59 women and 9 men, age 39.1 +/- 11.1 years; BMI 46.5 +/- 6.4 kg/m(2) (mean +/- SD)] were included. Fifty-six percent of patients had an abnormal Demester score, 44% had abnormal time spent at pH <4, and 80.9% had OSA [apnea hypopnea index (AHI) >10] and 39.7% had both conditions. The lower esophageal sphincter (LES) pressure was lower in patients with GERD (11.6 +/- 3.4 vs 13.4 +/- 3.6 mm Hg, respectively; P = 0.039). There was a relationship between AHI and BMI (r = 0.337; P = 0.005). Patients with OSA were older (40.5 +/- 10.9 vs 33.5 +/- 10.4 years; P = 0.039). GERD tended to be more frequent in patients with OSA (49.1% vs 23.1%, respectively; P = 0.089). There was no significant relationship between pH-metric data and AHI in either the 24-h total recording time or the nocturnal recording time. In multivariate analysis, GERD was significantly associated with a low LES pressure (P = 0.031) and with OSA (P = 0.045) but not with gender, age, and BMI. CONCLUSION: In this population of morbidly obese patients, OSA and GERD were frequent, associated in about 40% of patients. GERD was significantly associated with LES hypotonia and OSA independently of BMI. PMID- 18418661 TI - Clinical, genetic, molecular, and pathophysiological insights into spinocerebellar ataxia type 1. AB - Spinocerebellar ataxia type 1 (SCA1) is a late onset neurodegenerative disease characterized by cerebellar ataxia with variable degrees of ophthalmoplegia, pyramidal and extrapyramidal signs, and peripheral neuropathy. SCA1 is caused by the toxic effects triggered by an expanded polyglutamine (polyQ) within the protein ataxin 1 (Atxn1) resulting in variable degrees of neurodegeneration in the cerebellum, brainstem, and spinocerebellar tracts. The toxic gain-of-function mechanisms by which the polyQ expansion induces neuronal cell death are not fully understood and no effective therapies are yet available. Alterations in transcriptional regulation, calcium homeostasis, glutamate signaling/excitotoxicity, and impaired protein degradation are few recurrent events in the pathogenesis in SCA1. However, elucidating the molecular routes regulated by ataxin 1 is leading to the discovery of new pathways that are implicated in SCA1. This suggests that dominant-negative effects exerted by the mutant protein, rather than just gain-of-function mechanisms, might be also implicated in SCA1 pathogenesis. The challenge now is to determine how these responses account for the clinical manifestation of the disease symptoms and, ultimately, how this knowledge can be translated into the development of therapeutic strategies. Herein, we review the phenotype and most recent advances in our understanding of the physiopathological mechanisms of neurodegeneration in SCA1. PMID- 18418662 TI - Localization and functional roles of corticotropin-releasing factor receptor type 2 in the cerebellum. AB - The corticotropin-releasing factor (CRF) type 2 receptor has three splice variants alpha, beta, and gamma. In the rodent brain only CRFR2alpha is present. In the cerebellum, CRF-R2alpha has two different isoforms: a full-length form (fl) and truncated (tr). Both forms CRF-R2 have a unique cellular distribution. During postnatal cerebellar development, the expression patterns of tr and fl isoforms are changing. This suggests that, CRF and the related peptide urocortin (UCN) could play distinct roles in the immature and adult cerebellum, acting via different receptors subtypes. This review focuses on differences in the distribution of each isoform of CRF-R2 in view of their relationship to CRF and UCN release sites and their possible functional implications. Moreover, it includes novel findings of molecular pathways activating CRF-R2 isoforms through which CRF and UCN excert their specific actions. PMID- 18418663 TI - Expression, localization and possible actions of 25-Dx, a membraneassociated putative progesterone-binding protein, in the developing Purkinje cell of the cerebellum: a new insight into the biosynthesis, metabolism and multiple actions of progesterone as a neurosteroid. AB - Neurosteroids are now known to be steroids that are synthesized de novo from cholesterol in the central and peripheral nervous systems of vertebrates through mechanisms at least partly independent of peripheral steroidogenic glands, such as the adrenal and gonads. A series of our studies have demonstrated that the rat Purkinje cell, a cerebellar neuron, actively produces progesterone de novo from cholesterol only during neonatal life and progesterone promotes dendritic growth, spinogenesis and synaptogenesis via its nuclear receptor in this neuron. Thus the Purkinje cell serves as an excellent cellular model for understanding the formation of cerebellar neuronal circuit in relation to genomic neurosteroid actions. Recently, we have further found that Purkinje cells express the putative membrane progesterone receptor, 25-Dx in rats. By immunocytochemistry, the expression of 25-Dx was localized in the Purkinje cell and external granule cell layer. RT-PCR and Western immunoblot analyses revealed the expressions of 25-Dx and its mRNA in the rat cerebellum, which increased during neonatal life. Therefore, progesterone would promote dendritic growth, spinogenesis and synaptogenesis via 25-Dx as well as its nuclear receptor in the Purkinje cell in the neonate. Because the subcellular localization of 25-Dx was associated with membrane structures of the endoplasmic reticulum and Golgi, 25-Dx may also play a role in the regulation of neurosteroidogenesis in the developing Purkinje cell. Here we summarize the advances made in our understanding of the expression, localization and its possible actions of 25-Dx in the developing Purkinje cell. PMID- 18418664 TI - The roles of testicular orphan nuclear receptor 4 (TR4) in cerebellar development. AB - Since Testicular Receptor 4 (TR4) was cloned, efforts have been made to elucidate its physiological function. To examine the putative functions of TR4, the conventional TR4 knockout (TR4(-/-)) mouse model was generated. Throughout postnatal and adult stages, TR4(-/-) mice exhibited behavioral deficits in motor coordination, suggesting impaired cerebellar function. Histological examination of the postnatal and adult TR4(-/-) cerebellum revealed gross abnormalities in foliation. Further analyses demonstrated changes in the lamination of the TR4(-/ ) cerebellar cortex, including reduction in the thickness of both the molecular layer (ML) and the internal granule layer (IGL). Analyses of the developing TR4( /-) cerebellum indicate that the lamination irregularities observed may result from disrupted granule cell proliferation within the external granule cell layer (EGL), delayed inward migration of post-mitotic granule cells, and increased apoptosis during cerebellar development. In addition, abnormal development of Purkinje cells was observed in the postnatal TR4(-/-) cerebellum, as indicated by aberrant dendritic arborization. In postnatal, neuronal-specific TR4 knockout mice, architectural changes in the cerebellum were similar to those seen in TR4( /-) animals, suggesting that TR4 function in neuronal lineages might be important for cerebellar morphogenesis, and that the effect on Purkinje cell development is likely mediated by changes elsewhere, such as in granule cells, or is highly dependent on developmental stage. Together, our findings from various TR4 knockout mouse models suggest that TR4 is required for normal cerebellar development and that failure to establish proper cytoarchitecture results in dysfunction of the cerebellum and leads to abnormal behavior. PMID- 18418665 TI - Pharmacological characterization and anatomical distribution of the dopamine transporter in the mouse cerebellum. AB - We studied the binding parameters, the pharmacological profile and the anatomical distribution of the dopamine transporter in the mouse cerebellum by using the specific dopamine uptake antagonist [(3)H]GBR12935 and an antidopamine transporter monoclonal antibody. Competition experiments in cerebellar and striatal membrane preparations showed that [(3)H]GBR12935 binds to a specific binding site, sensitive to dopamine and low concentrations of mazindol. The affinity of dopamine for the cerebellar binding site was one order of magnitude lower than the affinity for the striatal binding site. Saturation experiments in cerebellar membrane preparations and thin frozen sections showed that the affinity of [(3)H]GBR12935 for this binding site is similar to its affinity for the striatal dopamine transporter. Saturable binding was lobule specific and in general was higher in the molecular layer compared to the granule cell layer. The immunohistochemical signal was mostly concentrated in the Purkinje cell layer and the cerebellar nuclei. The results suggest that the cerebellar dopamine transporter is similar but not identical to the striatal dopamine transporter and that it is present in the mouse cerebellum in a lobule and lamina specific pattern. PMID- 18418666 TI - Endocrine disrupting polyhalogenated organic pollutants interfere with thyroid hormone signalling in the developing brain. AB - Persistent polyhalogenated organic pollutants are present worldwide and accumulate along the food chain. They interfere with human and animal health and are particularly harmful for pre- and perinatal neurodevelopment. The mechanisms behind the observed effects vary depending on the specific compound investigated. Co-planar polychlorinated biphenyls (PCBs) can act via the arylhydrocarbon receptor while many ortho-substituted PCBs disrupt intracellular Ca(2+) homeostasis. A common mechanism for a wide variety of PCBs is interference with thyroid hormone (TH) signalling in developing brain, by changing intracellular TH availability or by interacting directly at the level of the TH receptors. Studies on gene expression in cortex and cerebellum revealed both hypothyroid- and hyperthyroid-like effects. However, since THdependent gene expression plays a crucial role in the coordination of neuronal proliferation, migration, synaptogenesis, myelination, etc., both reduced/delayed and increased/premature expression may result in permanent structural changes in neuronal communication networks, leading to lifelong deficits in cognitive performance, motor functions, and psychobehavior. In a similar way, PCBs are able to interfere with estrogen- and androgen-dependent brain development and in some studies neurobehavioral outcome was shown to be gender-specific. Other persistent organohalogens like polychlorinated dibenzo-p-dioxins (PCDDs) and polybrominated diphenyl ethers (PBDEs) also act as endocrine disrupters in the developing brain. Several of the mechanisms involved are similar to those of PCBs, but each group also works via own specific pathways. The fact that persistent organohalogens can amplify the neurotoxic effects of other environmental pollutants, such as heavy metals, further increases their risk for human and animal neurodevelopment. PMID- 18418668 TI - Thyrotropin-releasing hormone (TRH) in the cerebellum. AB - Thyrotropin-releasing hormone (TRH) was originally isolated from the hypothalamus. Besides controlling the secretion of TSH from the anterior pituitary, this tripeptide is widely distributed in the central nervous system and regarded as a neurotransmitter or modulator of neuronal activities in extrahypothalamic regions, including the cerebellum. TRH has an important role in the regulation of energy homeostasis, feeding behavior, thermogenesis, and autonomic regulation. TRH controls energy homeostasis mainly through its hypophysiotropic actions to regulate circulating thyroid hormone levels. Recent investigations have revealed that TRH production is regulated directly at the transcriptional level by leptin, one of the adipocytokines that plays a critical role in feeding and energy expenditure. The improvement of ataxic gait is one of the important pharmacological properties of TRH. In the cerebellum, cyclic GMP has been shown to be involved in the effects of TRH. TRH knockout mice show characteristic phenotypes of tertiary hypothyroidism, but no morphological changes in their cerebellum. Further analysis of TRH-deficient mice revealed that the expression of PFTAIRE protein kinase1 (PFTK1), a cdc2-related kinase, in the cerebellum was induced by TRH through the NO-cGMP pathway. The antiataxic effect of TRH and TRH analogs has been investigated in rolling mouse Nagoya (RMN) or 3 acetylpyridine treated rats, which are regarded as a model of human cerebellar degenerative disease. TRH and TRH analogs are promising clinical therapeutic agents for inducing arousal effects, amelioration of mental depression, and improvement of cerebellar ataxia. PMID- 18418667 TI - Mechanisms of ethanol-induced degeneration in the developing, mature, and aging cerebellum. AB - The adverse effects of acute and chronic ethanol exposure on cerebellar functions have been acknowledged for decades, in terms of impaired control of movement and balance. In addition to the motor impairment, cerebellar degeneration has recently been shown to contribute to distinct neuropsychological deficits in chronic alcoholics, as well as in children with prenatal ethanol exposure. The basic mechanisms underlying these ethanol-induced functional alterations and the related neuropathology in the cerebellum have mostly been clarified only recently. These mechanisms include: (i) excitotoxicity; (ii) dietary factors, especially thiamine depletion; (iii) glial abnormalities; (iv) changes in growth factors; (v) apoptotic mechanisms; (vi) oxidative stress; and (vii) compromised energy production. Although these mechanisms widely apply not only to the mature cerebellum, but also to the developing and the aging cerebella, the developing and the aged cerebellum have some special characteristics, which may make them even more vulnerable to ethanol-induced degeneration. These special instances will be discussed along with the general mechanisms of ethanol-induced cerebellar degeneration. PMID- 18418669 TI - Implications on cerebellar function from information coding. AB - One function of the cerebellar cortex is to process information. There are at least two types of information. Temporal information is encoded in the timing pattern of action and synaptic potentials, whereas structural information is encoded in the spatial pattern of the cerebellar synaptic circuitry. Intuitively, analysis of highly complex information in the time domain would require a cerebellar cortex with structural complexity to match. Information theory offers a way to estimate quantitatively both types of information and thereby helps to test hypotheses or advance theories of cerebellar neurobiology. These estimates suggest: (i) That the mossy-fiber-granule-cell system carries far more (temporal) information than the climbing fiber system, (ii) that Purkinje cells extract only a fraction of the (temporal) information from their afferents, and (iii) that the cerebellar cortex has a large (spatial) information coding capacity. Concerning information, one can argue that the cerebellar cortex analyzes temporal information in its afferents as a search engine, in search of coincidental mossy fiber events based on timing cues provided by climbing fiber events. Results of successive searches are continuously being converted into structural information encoded in the spatial distribution pattern of granule-cell-Purkinje-cell synapses along granule cell axons, thereby providing an adaptive and indeed self correcting dimension to the structural information code. The search engine operation involves cellular mechanisms acting on temporal events and is part of an associative learning process. The conversion and generation of structural information involves neuroplasticity mechanisms acting at the synaptic level, with electrophysiological as well as structural consequences, and may be part of the short- and long-term memory process. These and other attributes qualify the cerebellar cortex as a dynamic information processing center, contributing to memory and learning while linking motor output with sensory events. PMID- 18418670 TI - Functional asymmetry in the cerebellum: a brief review. AB - Recent discoveries on the way in which the cerebellum carries out higher non motor functions, have stimulated a proliferation of researches into functional integration and neural mechanisms in the cerebellum. Cerebellar functional asymmetry is a special characteristic of cerebellar functional organization and the cerebro-cerebellar circuitry that underlies task performance. Multi-level neuroimaging studies demonstrate that cerebellar functional asymmetry has a rather complex pattern, and may be correlated with practice or certain disorders. In this review, we summarize some new and important advances in the understanding of functional laterality of the cerebellum in primary motor and higher cognitive functions, and highlight the differences in the patterns of cerebellar functional asymmetry in the various functional domains. We propose that cerebellar functional asymmetry may be associated with the pattern of connectivity between a large number of widely distributed brain areas and between special cerebellar functional regions. It is suggested that cerebro-cerebellar circuits in particular play an important role in cerebellar functional asymmetry. Finally, we propose that multi-scale connectivity analyses and careful studies of high-level cerebellar functional asymmetry would make an important contribution to the understanding of the human cerebellum and cerebral neural networks. PMID- 18418671 TI - Compartmentation of GABA B receptor2 expression in the mouse cerebellar cortex. AB - Despite the apparent uniformity in cellular composition of the adult mammalian cerebellar cortex, it is actually highly compartmentalized into transverse zones, and within each zone the cortex is further subdivided into a reproducible array of parasagittal stripes. The most extensively studied compartmentation antigen is zebrin II/aldolase c, which is expressed by a subset of Purkinje cells forming parasagittal stripes. Gamma-aminobutyric acid B receptors (GABABRs) are G-protein coupled receptors that mediate a slow, prolonged form of inhibition in many brain areas. This study examines the localization of GABABR2 in the mouse cerebellum by using whole mount and section immunohistochemistry. The data reveal that GABABR2 immunoreactivity is expressed strongly in the dendrites of a subset of Purkinje cells that form a reproducible array of transverse zones and parasagittal stripes. By using double immunostaining, the striped pattern of GABABR2 expression was shown to be identical to that revealed by anti-zebrin II and complementary to that of phospholipase Cbeta4. This finding supports previous functional studies showing that inhibitory neurotransmission is highly patterned in the cerebellar cortex. PMID- 18418672 TI - Steroids, sex and the cerebellar cortex: implications for human disease. AB - Neurosteroids play an important role in the development of the cerebellum. In particular, estradiol and progesterone appear capable of inducing increases in dendritic spine density during development, and there is evidence that both are synthesized de novo in the cerebellum during critical developmental periods. In normal neonates and adults, there are few differences in the cerebellum between the sexes and most studies indicate that hormone and receptor levels also do not differ significantly during development. However, the sexes do differ significantly in risk of neuropsychological diseases associated with cerebellar pathology, and in animal models there are noticeable sex differences in the response to insult and genetic mutation. In both humans and animals, males tend to fare worse. Boys are more at risk for autism and Attention Deficit Hyperactivity Disorder than girls, and schizophrenia manifests at an earlier age in men. In rats males fare worse than females after perinatal exposure to polychlorinated biphenyls, and male mice heterozygous for the staggerer and reeler mutation show a more severe phenotype. Although very recent evidence suggests that differences in neurosteroid levels between the sexes in diseased animals may play a role in generating different disease phenotypes, the reason this hormonal difference occurs in diseased but not normal animals is currently unknown. PMID- 18418673 TI - Thyroid hormone transporters in the brain. AB - Thyroid hormone plays an essential role in proper mammalian development of the central nervous system and peripheral tissues. Lack of sufficient thyroid hormone results in abnormal development of virtually all organ systems, a syndrome termed cretinism. In particular, hypothyroidism in the neonatal period causes serious damage to neural cells and leads to mental retardation. Although thyroxine is the major product secreted by the thyroid follicular cells, the action of thyroid hormone is mediated mainly through the deiodination of T(4) to the biologically active form 3,3', 5-triiodo-L-thyronine, followed by the binding of T(3) to a specific nuclear receptor. Before reaching the intracellular targets, thyroid hormone must cross the plasma membrane. Because of the lipophilic nature of thyroid hormone, it was thought that they traversed the plasma membrane by simple diffusion. However, in the past decade, a membrane transport system for thyroid hormone has been postulated to exist in various tissues. Several classes of transporters, organic anion transporter polypeptide (oatp) family, Na(+)/Taurocholate cotransporting polypeptide (ntcp) and amino acid transporters have been reported to transport thyroid hormones. Monocarboxylate transporter8 (MCT8) has recently been identified as an active and specific thyroid hormone transporter. Mutations in MCT8 are associated with severe X-linked psycomotor retardation and strongly elevated serum T3 levels in young male patients. Several other molecules should be contributed to exert the role of thyroid hormone in the central nervous system. PMID- 18418674 TI - Sporadic ataxias in Japan--a population-based epidemiological study. AB - Sporadic spinocerebellar ataxias (SCAs) comprise heterogeneous diseases with poorly understood epidemiologies and etiologies. A population-based epidemiological analysis of sporadic ataxias in the Japanese population was described. The prevalence rate of SCAs in the Japanese population is estimated to be 18.5/100,000. Sporadic SCAs account for 67.2% of total SCAs including hereditary SCAs, with olivopontocerebellar atrophy (OPCA) being the most common form sporadic ataxia (64.7%). The natural history analysis conducted on the basis of International Cooperative Ataxia Rating Scale (ICARS) showed that only 33% of patients with OPCA were able to walk at least with one stick 4-5 years after the onset of OPCA, which is much less than that of patients with cortical cerebellar atrophy (CCA). Similarly, 43% of patients with OPCA were able to stand alone 4-5 years after the onset, while 76% of patients with CCA were able to stand alone at the same disease duration. A population-based epidemiological analysis should provide essential information on the natural history of SCAs. PMID- 18418676 TI - Spinocerebellar ataxias caused by polyglutamine expansions: a review of therapeutic strategies. AB - Six of the spinocerebellar ataxias (SCAs) are caused by expanded CAG trinucleotide repeats encoding polyglutamine tracts in different genes. Together with three other neurodegenerative diseases they represent the polyglutamine repeat disorders. These disorders share many pathological features beyond a common genetic mechanism. They are the subject of considerable research efforts to elucidate their basic pathophysiologies, with the hope of using this knowledge to develop disease modifying treatments. Here we examine the biology that underpins possible therapeutic strategies for the SCAs caused by CAG repeats and review supportive data from cell and animal models. Therapeutic strategies include silencing gene expression, increasing protein clearance, reducing the toxicity of the protein, influencing downstream pathways activated by the mutant protein and transplantation. We also consider strategies which have been tested in other polyglutamine diseases that may generalize to these SCAs. Finally, we review clinical trials and consider the problems of translating the increasing amount of promising laboratory data into human trials. PMID- 18418675 TI - Molecular pathogenesis and cellular pathology of spinocerebellar ataxia type 7 neurodegeneration. AB - Spinocerebellar ataxia type 7 (SCA7) is unique among CAG/polyglutamine (polyQ) repeat diseases due to dramatic intergenerational instability in repeat length and an associated cone-rod dystrophy retinal degeneration phenotype. SCA7 is caused by a polyQ expansion in the protein ataxin-7. Like other neurodegenerative diseases caused by polyQ expansion mutations, the spectrum of clinical severity and disease progression worsens with increasing polyQ length. Several potential mechanisms for the molecular pathogenesis of polyQ-expanded ataxin-7 have been suggested. These include, but are not limited to, alteration of endogenous ataxin 7 function, abnormal processing and stability of polyQ ataxin-7, and alteration of transcriptional regulation via interaction of polyQ-expanded ataxin-7 with other transcriptional regulators. Ataxin-7's normal function as a transcription factor may contribute to the selective vulnerability of specific cellular populations in SCA7, and the resolution of the mechanistic basis of this pathogenic cascade is a major focus of SCA7 disease research. PolyQ-expanded ataxin-7 can cause non-cell autonomous neurodegeneration in cerebellar Purkinje cells. Advances in understanding SCA7's molecular basis have led to important insights into cell-type specific neurodegeneration. We expect that further study of ataxin-7 normal function, insights into the molecular basis of SCA7 neurodegeneration, and the development of therapeutic interventions for SCA7 will greatly influence related endeavors directed at other CAG/polyQ repeat diseases. PMID- 18418677 TI - Magnetic resonance imaging in spinocerebellar ataxias. AB - Magnetic resonance (MR) imaging is widely used to visualize atrophic processes that occur during the pathogenesis of spinocerebellar ataxias (SCAs). T1-weighted images are utilized to rate the atrophy of cerebellar vermis, cerebellar hemispheres, pons and midbrain. Signal changes in the basal ganglia and ponto cerebellar fibers are evaluated by T2-weighted and proton density-weighted images. However, two-dimensional (2D) images do not allow a reliable quantification of the degree of atrophy. The latter is now possible through the application of three-dimensional (3D) true volumetric methods, which should be used for research purposes. Ideally, these methods should allow automated segmentation of contrast-defined boundaries by using region growing algorithms, which can be applied successfully in structures of the posterior fossa and basal ganglia. Thin slice thickness helps to minimize partial volume effects. Whereas volumetric approaches rely on predetermined anatomical boundaries, voxel-based morphometry has been developed to determine group differences between different types of SCA (cross-sectional studies) or within one SCA entity (longitudinal studies). We will review recent results and how these methods are currently used to (i) separate sporadic and dominantly inherited forms of cerebellar ataxias; (ii) identify specific SCA genotypes; (iii) correlate patho-anatomical changes with SCA disease symptoms or severity; and (iv) visualize and estimate the rate of progression in SCA. PMID- 18418678 TI - Electrophysiology in spinocerebellar ataxias: spread of disease and characteristic findings. AB - Spinocerebellar ataxias (SCAs) comprise a clinically and genetically heterogeneous group of autosomal dominantly inherited neurodegenerative disorders affecting the cerebellum and to variable degrees further parts of the nervous system. Electrophysiology is a potent tool to prove impairment of multiple neuronal systems and fibre tracts and even to decipher subclinical affection. Electrooculography, evoked potentials, nerve conduction studies and polysomnography are especially helpful in the setting of SCAs. Severely slowed saccades are a hallmark of SCA2. Vertical nystagmus occurs most frequently in SCA3 and SCA6. Visual potentials recede especially in SCA7. Substantially prolonged central motor conduction times in motor-evoked potentials occur frequently in SCA1 even in patients without clinical signs of pyramidal affection. Thus, electrophysiological analyses may help to predict the SCA genotype and direct molecular genetic diagnostics. Polysomnography is a helpful tool in the analysis of sleep disorders and frequently helps to decipher treatable causes like periodic leg movement in sleep and REM sleep behaviour disorder in SCAs. Nerve conduction studies reveal sensory neuropathy in all common SCA subtypes, but to variable degrees. Age rather than CAG repeat length appears to be the most important determinant for neuropathy and makes sensory nerve action potentials a potential progression marker in SCA. PMID- 18418679 TI - The clinical diagnosis of autosomal dominant spinocerebellar ataxias. AB - The spinocerebellar ataxias (SCAs) are a heterogeneous group of autosomal dominantly inherited progressive ataxia diseases. Up to now, almost 30 different gene loci have been found. In 14 of them, the underlying mutations have been identified. The more common SCAs, SCA1, 2, 3 and 6 are due to translated CAG repeat expansions that code for an elongated polyglutamine tract within the respective proteins. These diseases belong to a larger group of polyglutamine disorders that also includes Huntington's disease. Epidemiological studies conducted in different European regions found prevalence rates of SCAs ranging from 0.9 to 3.0:100,000. In all SCAs, ataxia is the prominent symptom. However, the majority have a complex phenotype in which ataxia is accompanied by varying non-ataxia symptoms. In all ataxia patients with proven or suspected autosomal dominant mode of inheritance, the available molecular genetic tests for SCA mutations should be performed. Depending on the geographical origin of the family, these tests will lead to positive diagnostic results in at least half of the families. PMID- 18418680 TI - Clinical and genetic analysis of spinocerebellar ataxia type 11. AB - The autosomal dominant cerebellar ataxias (ADCAs) are a genetically heterogeneous group of disorders. Clinical classification of the ADCAs into three types has facilitated defining phenotypes and in turn, linkage analysis, which has led to the discovery of 30 loci and 16 genes. The type III ADCAs are 'pure' spinocerebellar ataxias (SCA), those that appear to elude neurological features outside of the cerebellum. At present 3 ADCA type III SCA genes have been published, SCA5, SCA6, and SCA14, these three genes appear to have various roles suggesting involvement in both different and possibly overlapping neurodegenerative pathways. The known ADCAIII genes are thought to have such roles as involvement in signal transduction, cell proliferation, synaptic transmission, and channel regulation. Here we update readers on the current progress on SCA11 and the identification of the disease gene. We discuss the clinical, genetic, and pathological details of SCA11--a locus at chromosome 15q14 q21.3 in a Caucasian family of British ancestry. We also discuss the refining of this region, and methods used to prioritize the screening of the over 130 candidate genes in this genomic region. PMID- 18418682 TI - The highly heterogeneous spinocerebellar ataxias: from genes to targets for therapeutic intervention. PMID- 18418683 TI - Fenestration of the posterior inferior cerebellar artery. AB - Fenestration of the posterior inferior cerebellar artery (PICA) is exceedingly rare. Only one known example - a right PICA fenestration, has been documented in the peer-reviewed literature. A left-sided PICA fenestration is presented in this unique case illustration. PMID- 18418681 TI - Thyroid hormone and cerebellar development. AB - Thyroid hormone (TH) plays a key role in mammalian brain development. The developing brain is sensitive to both TH deficiency and excess. Brain development in the absence of TH results in motor skill deficiencies and reduced intellectual development. These functional abnormalities can be attributed to maldevelopment of specific cell types and regions of the brain including the cerebellum. TH functions at the molecular level by regulating gene transcription. Therefore, understanding how TH regulates cerebellar development requires identification of TH-regulated gene targets and the cells expressing these genes. Additionally, the process of TH-dependent regulation of gene expression is tightly controlled by mechanisms including regulation of TH transport, TH metabolism, toxicologic inhibition of TH signaling, and control of the nuclear TH response apparatus. This review will describe the functional, cellular, and molecular effects of TH deficit in the developing cerebellum and emphasize the most recent findings regarding TH action in this important brain region. PMID- 18418684 TI - Spinocerebellar ataxia 2 (SCA2). AB - Spinocerebellar ataxia type 2 (SCA2) is an autosomal dominantly inherited, neurodegenerative disease. It can manifest either with a cerebellar syndrome or as Parkinson's syndrome, while later stages involve mainly brainstem, spinal cord and thalamus. This particular atrophy pattern resembles sporadic multi-system atrophy (MSA) and results in some clinical features indicative of SCA2, such as early saccade slowing, early hyporeflexia, severe tremor of postural or action type, and early myoclonus. For treatment, levodopa is temporarily useful for rigidity/bradykinesia and for tremor, magnesium for muscle cramps, but neuroprotective therapy will depend on the elucidation of pathogenesis. The disease cause lies in the polyglutamine domain of the protein ataxin-2, which can expand in families over successive generations resulting in earlier onset age and faster progression. Genetic testing in SCA2 and other polyglutamine disorders like the well-studied Huntington's disease is now readily available for family planning. Although these disorders differ clinically and in the affected neuron populations, it is not understood how the different polyglutamine proteins mediate such tissue specificity. The neuronal intranuclear inclusion bodies described in other polyglutamine disorders are not frequent in SCA2. For the quite ubiquitously expressed ataxin-2, a subcellular localization at the Golgi, the endoplasmic reticulum and the plasma membrane, in interaction with proteins of mRNA translation and of endocytosis have been observed. As a first victim of SCA2 degeneration, cerebellar Purkinje neurons may be preferentially susceptible to alterations of these subcellular pathways, and therefore our review aims to portray the particular profile of the SCA2 disease process and correlate it to the specific features of ataxin-2. PMID- 18418685 TI - An overview of nuclear receptor coregulators involved in cerebellar development. AB - Nuclear receptors (NRs) precisely control the gene regulation throughout the development of the central nervous system, including the cerebellum. Functionally, the full activity of NRs requires their cognate coregulators to be recruited by NRs and modulate the activation or repression of target gene expression. Recent progress of in vitro studies of NR coregulators has revealed that NR coregulators form large complexes in a cyclic manner and subsequently exert genetic and epigenetic influence via various intrinsic enzyme activities. Moreover, NR coregulators physiologically provide a combinatorial code for time- and gene-specific responses depending on their expression levels, relative affinities for individual receptors, and posttranslational modification. Since expression of many cerebellar genes is known to be regulated by NRs critical in a specific period for cerebellar development, their partnership with cognate coregulators may be an important factor for normal cerebellar development. This review summarizes current findings regarding the molecular structures, molecular mechanisms, temporal and spatial expression patterns, and possible biological functions of NR coregulators related to cerebellar development. PMID- 18418688 TI - Slowly progressive spinocerebellar ataxia with extrapyramidal signs and mild cognitive impairment (SCA21). AB - Spinocerebellar ataxia 21 is a slowly progressive and mild ataxia associated with extrapyramidal signs. Affected subjects exhibit a moderate gait and limb ataxia variably associated with akinesia, tremor, rigidity, hyporeflexia, and mild cognitive impairment. The responsible gene has been assigned to a 19 Mbases interval on chromosome 7p in a single French family. No evidence of significant linkage to this locus was found in 21 other families obtained from the EUROSCA consortium. The locus interval contains several candidate genes that could be responsible for the disease. Direct sequencing of NDUFA4, PHF14, KIAA0960, ARLA4, ETV1, DGKB, HDAC9, FERD3L, ITGB8, and SP4 genes were performed, but all the direct mutation analyses were negative excluding pathogenic mutations associated with the disease. Therefore, the gene responsible for SCA21 remains to be identified. PMID- 18418686 TI - The role of cerebellar genes in pathology of autism and schizophrenia. AB - Schizophrenia and autism are neurodevelopmental diseases that have genetic as well as environmental etiologies. Both disorders have been associated with prenatal viral infection. Brain imaging and postmortem studies have found alterations in the structure of the cerebellum as well as changes in gene expression. Our laboratory has developed an animal model using prenatal infection of mice with human influenza virus that has demonstrated changes in behavior, pharmacology, structure, and gene expression in the brains of exposed offspring. In the current communication we describe altered expression of cerebellar genes associated with development of brain disorder in a mouse model for schizophrenia and autism and correlate these changes with those involved in the pathology of these two disorders. PMID- 18418687 TI - Spinocerebellar ataxia 17 (SCA17) and Huntington's disease-like 4 (HDL4). AB - Spinocerebellar ataxia 17 (SCA17) or Huntington's disease-like-4 is a neurodegenerative disease caused by the expansion above 44 units of a CAG/CAA repeat in the coding region of the TATA box binding protein (TBP) gene leading to an abnormal expansion of a polyglutamine stretch in the corresponding protein. Alleles with 43 and 44 repeats have been identified in sporadic cases and their pathogenicity remains uncertain. Furthermore, incomplete penetrance of pathological alleles with up to 49 repeats has been suggested. The imperfect nature of the repeat makes intergenerational instability extremely rare and de novo mutations are most likely the result of partial duplications. This is one of the rarer forms of autosomal dominant cerebellar ataxia but the associated phenotype is often severe, involving various systems (cerebral cortex, striatum, and cerebellum), with extremely variable age at onset (range: 3-75 years) and clinical presentation. This gene is thought to account for a small proportion of patients with a Huntington's disease-like phenotype and cerebellar signs. Parkinson's disease-like, Creutzfeldt-Jakob disease-like and Alzheimer disease like phenotypes have also been described with small SCA17 expansions. The abnormal protein is expressed at the same level as its normal counterpart and forms neuronal intranuclear inclusions containing other proteins involved in protein folding or degradation. The increase in the size of the glutamine stretch enhances transcription in vitro, probably leading to transcription deregulation. Interestingly, the TBP protein mutated in SCA17 is recruited in the inclusions of other polyglutaminopathies, suggesting its involvement in the transcription down regulation observed in these diseases. PMID- 18418689 TI - SCA3: neurological features, pathogenesis and animal models. AB - The most frequent subtype of autosomal dominant inherited spinocerebellar ataxias is caused by CAG repeat expansions of more than 55 units in the ataxin-3 gene. The clinical variability of the phenotype depends on the length of the expanded repeat and the age at onset (and thus indirectly with the repeat size). Anticipation of the phenotype is most frequently associated with repeat expansions in paternal transmission. In this review we describe four clinical subphenotypes and correlate them to the respective repeat expansions. We also provide a detailed description of the neuropathological features. Finally, we discuss the current knowledge on the function of normal and dysfunction of altered ataxin-3 and how this translates to the predicted structure of the protein. PMID- 18418690 TI - Progress in transduction of cerebellar Purkinje cells in vivo using viral vectors. AB - Expression of a foreign gene in cerebellar Purkinje cells in vivo is a powerful method for exploring the pathophysiology of the cerebellum. Although using developmental engineering many gene-modified mice have been generated, this approach is time-consuming and requires a lot of effort for crossing different lines of mice, genotyping and maintenance of animals. If a gene of interest can be transferred to and efficiently expressed in Purkinje cells of developing and mature animals, it saves much time, effort and money. Recent advances in viral vectors have markedly contributed to selective and efficient gene transfer to Purkinje cells in vivo. There are two approaches for selective gene expression in Purkinje cells: one is to take advantage of the viral tropism for Purkinje cells, which includes the tropism of adeno-associated virus and the vesicular stomatitis virus glycoprotein (VSV-G)-pseudotyped lentivirus. Another method, which might be used in combination with the first one, is utilization of a Purkinje-cell specific promoter. Focusing mainly on these points, recent progress in viral vector-mediated transduction of Purkinje cells in vivo is reviewed. PMID- 18418691 TI - Human cerebellar responses to brush and heat stimuli in healthy and neuropathic pain subjects. AB - Though human pain imaging studies almost always demonstrate activation in the cerebellum, the role of the cerebellum in pain function is not well understood. Here we present results from two studies on the effects of noxious thermal heat and brush applied to the right side of the face in a group of healthy subjects (Group I) and a group of patients with neuropathic pain (Group II) who are more sensitive to both thermal and mechanical stimuli. Statistically significant activations and volumes of activations were defined in the cerebellum. Activated cerebellar structures were identified by colocalization of fMRI activation with the 'MRI Atlas of the Human Cerebellum'. Functional data (obtained using a 3T magnet) were defined in terms of maximum voxels and volume of activation in the cerebellum. Volume maps were then mapped onto two millimeter serial slices taken through the cerebellum in order to identify activation within regions defined by the activation volume. The data indicate that different regions of the cerebellum are involved in acute and chronic pain processing. Heat produces greater contralateral activation compared with brush, while brush resulted in more ipsilateral/bilateral cerebellar activation. Further, innocuous brush stimuli in healthy subjects produced decreased cerebellar activation in lobules concerned with somatosensory processing. The data also suggest a dichotomy of innocuous stimuli/sensorimotor cerebellum activation versus noxious experience/cognitive/limbic cerebellum activation. These results lead us to propose that the cerebellum may modulate the emotional and cognitive experience that distinguishes the perception of pain from the appreciation of innocuous sensory stimulation. PMID- 18418692 TI - Bidirectional expression of the SCA8 expansion mutation: one mutation, two genes. AB - Spinocerebellar ataxia type 8 (SCA8) is a dominantly inherited, slowly progressive neurodegenerative disorder caused by a CTG.CAG repeat expansion located on chromosome 13q21. The expansion mutation was isolated directly from the DNA of a single patient using RAPID cloning and subsequently shown to co segregate with disease in additional ataxia families including a seven-generation kindred (the MN-A family). The size-dependent penetrance of the repeat found in the large MN-A kindred makes it appear as though some parts of the family have a dominant disorder while other parts of this same family have recessive or sporadic forms of ataxia. While the linkage and size-dependent penetrance of the SCA8 CTG.CAG expansion in the MN-A family argue that the SCA8 expansion causes ataxia, the reduced penetrance in other SCA8 families and the discovery of expansions in the general population have led to a controversy surrounding whether or not the SCA8 expansion is pathogenic. A recently reported mouse model in which SCA8 BAC-expansion but not BAC-control lines develop a progressive neurological phenotype now demonstrates the pathogenicity of the (CTG.CAG)(n) expansion. These mice show a loss of cerebellar GABAergic inhibition and, similar to human patients, have 1C2-positive intranuclear inclusions in Purkinje cells and other neurons. Additional studies demonstrate that the SCA8 expansion is expressed in both directions (CUG and CAG) and that a novel gene expressed in the CAG direction encodes a pure polyglutamine expansion protein (ataxin 8, ATXN8). Moreover, the expression of non-coding (CUG)(n) expansion transcripts (ataxin 8 opposite strand, ATXN8OS) and the discovery of intranuclear polyglutamine inclusions suggest SCA8 pathogenesis may involve toxic gain-of-function mechanisms at both the protein and RNA levels. Our data, combined with the recently reported antisense transcripts spanning the DM1 repeat expansion in the CAG direction and the growing number of reports of antisense transcripts expressed throughout the mammalian genome, raises the possibility that bidirectional expression across pathogenic microsatellite expansions may occur in other expansion disorders, and that potential pathogenic effects of mutations expressed from both strands should be considered. PMID- 18418694 TI - Serum cystatin C measured by a sol particle homogeneous immunoassay can accurately detect early impairment of renal function. AB - BACKGROUND: A sol particle homogeneous immunoassay (SPIA) is a method to measure the serum cystatin C (cysC) level as a marker of the glomerular filtration rate (GFR). Recently, formulas to convert measured cysC to GFR have been developed. METHODS: A total of 154 patients (46 +/- 18 years old) who had undergone renal biopsy, sodium thiosulfate clearance (C thio) and 24-h creatinine clearance (24 hCcr) tests were subjects for the study. Their serum cysC levels were determined by SPIA. RESULTS: Multiple regression analysis revealed C (thio) and age as independent variables for serum creatinine concentration (Cr), while only C thio affected cysC. The equations using Cr or cysC showed significant correlation with C thio. Receiver-operating curve ROC analysis revealed that cysC and 24-hCcr shared comparable power to detect patients with GFR < 90 or 60 ml/min/1.73 m2 (AUC = 0.862 and 0.943 vs. AUC = 0.842 and 0.943, respectively), while Cr (AUC = 0.881) and MDRD2 (AUC = 0.888) showed slightly inferior ability to detect 60 ml/min/1.73 m2 than other parameters in the female population. The cut-off point of cysC and Cr obtained from the ROC analysis demonstrated strong power to detect patients with C thio < 90 ml/min/1.73 m2 or C thio < 60 ml/min/1.73 m2. According to CKD stages, the mean values of each equation were significantly different, like that demonstrated by 24-hCcr. CONCLUSION: SPIA could determine cysC levels that detected early renal impairment. The accuracy of cysC to detect early renal impairment may be superior to that of Cr in females, while it would be comparable to that of CG or MDRD when they are corrected by sex and age. Both cysC itself and cysC equations are effective to monitor the progress of renal impairment. The future standardization of cysC measurements and development of novel equation of cysC would contribute to the further improvement of GFR estimation in clinical practice. PMID- 18418696 TI - Inhibitory effect of the rhizomes of Alpinia officinarum on TPA-induced inflammation and tumor promotion in two-stage carcinogenesis in mouse skin. AB - The methanol extract of galangal (the rhizomes of Alpinia officinarum L.) exhibited remarkable antitumor-promoting activity on an in vivo two-stage carcinogenesis test of mice using 7,12-dimethylbenz[a]anthracene as an initiator and 12-O-tetradecanoylphorbol-13-acetate (TPA) as a promoter. Seven diarylheptanoids (1-7) were isolated and identified from the active fraction of the methanol extracts of the galangal. These compounds, 1-7, were evaluated for their inhibitory effects on TPA-induced inflammation (1 microg/ear) in mice. These compounds (1-7) tested showed marked anti-inflammatory effects, with a 50% inhibitory dose of 0.8-2.7 micromol/ear. PMID- 18418695 TI - The role of cell death in the pathogenesis of autoimmune disease: HMGB1 and microparticles as intercellular mediators of inflammation. AB - Cell death is critical to normal homeostasis, although this process, when increased aberrantly, can lead to the production of pro-inflammatory mediators promoting autoimmunity. Two novel intercellular mediators of inflammation generated during cell death are high mobility group box 1 (HMGB1) protein and microparticles (MPs). HMGB1 is a nuclear protein that functions in transcription when inside the nucleus but takes on pro-inflammatory properties when released during cell death. Microparticles are small, membrane-bound structures that extrude from cells when they die and contain cell surface proteins and nuclear material from their parent cells. MPs circulate widely throughout the vasculature and mediate long-distance communication between cells. Both MPs and HMGB1 have been implicated in the pathogenesis of a broad spectrum of inflammatory diseases, including the prototypic autoimmune conditions systemic lupus erythematosus and rheumatoid arthritis. Given their range of activity and association with active disease, both structures may prove to be targets for effective therapy in these and other disorders. PMID- 18418693 TI - Brain development, environment and sex: what can we learn from studying graviperception, gravitransduction and the gravireaction of the developing CNS to altered gravity? AB - As man embarks on space exploration and contemplates space habitation, there is a critical need for basic understanding of the impact of the environmental factors of space, and in particular gravity, on human survival, health, reproduction and development. This review summarizes our present knowledge on the effect of altered gravity on the developing CNS with respect to the response of the developing CNS to altered gravity (gravireaction), the physiological changes associated with altered gravity that could contribute to this effect (gravitransduction), and the possible mechanisms involved in the detection of altered gravity (graviperception). Some of these findings transcend gravitational research and are relevant to our understanding of the impact of environmental factors on CNS development on Earth. PMID- 18418697 TI - Antitumor actions of a chromone glucoside cnidimoside A isolated from Cnidium japonicum. AB - In a series of studies on the search for new antitumor and antimetastatic substances from the natural medicinal plants of the Umbelliferae family, we previously reported that chalcone derivatives isolated from Angelica keisekei roots have antitumor and antimetastatic activities. In the present study, we examined the effects of a chromone glucoside cnidimoside A isolated from Cnidium japonicum whole plants on tumor growth and tumor metastasis in colon 26-bearing mice. Cnidimoside A (50 mg/kg, twice daily) significantly inhibited tumor growth and final tumor weight compared to the growth in vehicle-treated colon 26-bearing mice (control). Furthermore, the number of mice with abdominal invasion of tumors was also reduced by orally administered cnidimoside A (50 mg/kg, twice daily). In this study, the CD8(+) T Cell- and interferon (IFN)-gamma-positive cell numbers in the small intestine in the colon 26-bearing mice were significantly reduced compared with those in the normal mice, but the natural killer (NK)-positive cell number did not differ significantly between the normal and colon 26-bearing mice. The CD8(+) T-, NK and IFN-gamma-positive cell numbers in the small intestine were significantly increased by orally administered cnidimoside A (50 mg/kg, twice daily) compared to those in vehicle-treated colon 26-bearing mice. In conclusion, it seems likely that the antitumor and antimetastatic actions of cnidimoside A may be partly associated with the stimulation of immune response in the small intestine. PMID- 18418698 TI - Fludarabine/intermediate-dose cytarabine with or without allogeneic hematopoietic stem cell transplantation in poor-risk leukemia: a single center experience. AB - Disease recurrence has been and remains the leading cause of treatment failure in patients with high-risk leukemia. We retrospectively analyzed outcome in 61 patients with high-risk leukemia receiving a combination of fludarabine and intermediate-dose cytarabine as induction (n = 11) or salvage therapy (n = 35). Thirty-six patients having a suitable stem cell donor proceeded to allogeneic hematopoietic stem cell transplantation (HSCT). Ten patients received fludarabine based salvage therapy without consecutive allogeneic transplantation and 15 patients received fludarabine/intermediate-dose cytarabine because of disease relapse following allogeneic stem cell transplantation. In patients without prior allogeneic HSCT (n = 46) the complete remission rate (CR) was 41% with a CR rate of 46 and 14% in patients with acute myeloid leukemia (AML) and with acute lymphoblastic leukemia (ALL), respectively. Overall survival for patients achieving a CR was 41 versus 0% for patients not achieving CR (P < 0.0001). The best outcome was observed in patients receiving an allogeneic HSCT in CR following fludarabine/ intermediate-dose cytarabine (47 vs. 0% for patients not in CR at the time of allografting, P = 0.01). All 10 patients receiving fludarabine/intermediate-dose cytarabine without subsequent allogeneic HSCT died within 3 years either of disease relapse/progression or infection. Only 1/15 (7%) patients receiving fludarabine/intermediate-dose cytarabine because of relapse following allogeneic HSCT became a long-term survivor. By multivariate analysis achieving CR, receiving an allogeneic HSCT, and being in first relapse or untreated were the only parameters that significantly determine the outcome. Although preliminary only high-risk AML patients having a stem cell donor are candidates for fludarabine/intermediate-dose cytarabine and only those achieving a CR should be referred to subsequent allogeneic HSCT. All other patients with high-risk leukemia are candidates for experimental therapies within controlled trials. PMID- 18418699 TI - Abnormal Igf2 gene in Prague hereditary hypertriglyceridemic rats: its relation to blood pressure and plasma lipids. AB - Prague hypertriglyceridemic (HTG) rats represent a suitable model of metabolic syndrome. We have established the set of F(2) hybrids derived from HTG and Lewis progenitors to investigate the relationship between respective polymorphism(s) of Igf2 gene and blood pressure (BP) or other cardiovascular phenotypes. HTG rats had elevated systolic BP and plasma triglycerides but lower plasma cholesterol compared to Lewis rats of both genders. In males, there was higher mean arterial pressure, diastolic BP and relative heart weight in HTG than in Lewis rats. The results obtained in the total population of F(2) hybrids indicated strong segregation of Igf2 genotype with plasma triglycerides. There was no segregation of Igf2 genotype with any BP component except BP changes occurring after the blockade of either renin-angiotensin system (RAS) or NO synthase. When F(2) population was analyzed according to gender, male F(2) progeny homozygous for HTG Igf2 allele had significantly higher plasma triglycerides and greater BP changes after NO synthase blockade than those homozygous for Lewis allele. On the contrary, male F(2) progeny homozygous for HTG Igf2 allele had significantly lower plasma cholesterol and smaller BP changes after RAS blockade. PCR analysis of Igf2 gene by using of microsatelite D1Mgh22 has shown polymorphism between HTG and Lewis rats. Sequence analysis of cDNA revealed insertion of 14 nucleotides in HTG gene. In conclusion, polymorphism in Igf2 gene may be responsible for differences in lipid metabolism between HTG and Lewis rats. It remains to determine how these abnormalities could be involved in BP regulation by particular vasoactive systems. PMID- 18418700 TI - Modulation of the c-Jun N-terminal kinase activity in the embryonic heart in response to anoxia-reoxygenation: involvement of the Ca2+ and mitoKATP channels. AB - Whether the response of the fetal heart to ischemia-reperfusion is associated with activation of the c-Jun N-terminal kinase (JNK) pathway is not known. In contrast, involvement of the sarcolemmal L-type Ca2+ channel (LCC) and the mitochondrial KATP (mitoKATP) channel has been established. This work aimed at investigating the profile of JNK activity during anoxia-reoxygenation and its modulation by LCC and mitoK(ATP) channel. Hearts isolated from 4-day-old chick embryos were submitted to anoxia (30 min) and reoxygenation (60 min). Using the kinase assay method, the profile of JNK activity in the ventricle was determined every 10 min throughout anoxia-reoxygenation. Effects on JNK activity of the LCC blocker verapamil (10 nM), the mitoK(ATP) channel opener diazoxide (50 microM) and the blocker 5-hydroxydecanoate (5-HD, 500 microM), the mitochondrial Ca2+ uniporter (MCU) inhibitor Ru360 (10 microM), and the antioxidant N-(2 mercaptopropionyl) glycine (MPG, 1 mM) were determined. In untreated hearts, JNK activity was increased by 40% during anoxia and peaked fivefold relative to basal level after 30-40 min reoxygenation. This peak value was reduced by half by diazoxide and was tripled by 5-HD. Furthermore, the 5-HD-mediated stimulation of JNK activity during reoxygenation was abolished by diazoxide, verapamil or Ru360. MPG had no effect on JNK activity, whatever the conditions. None of the tested pharmacological agents altered JNK activity under basal normoxic conditions. Thus, in the embryonic heart, JNK activity exhibits a characteristic pattern during anoxia and reoxygenation and the respective open-state of LCC, MCU and mitoKATP channel can be a major determinant of JNK activity in a ROS-independent manner. PMID- 18418702 TI - Association of occupational and non-occupational risk factors with the prevalence of work related carpal tunnel syndrome. AB - PURPOSE: Several occupational and personal risk factors cause the development of carpal tunnel syndrome (CTS). The purpose of the study was to evaluate both non occupational and occupational factors associated with CTS in industrial workers. METHODS: A cross sectional study was designed with 400 industrial workers (77% male, 23% female) randomly selected. Workers' upper extremities were examined and related signs and symptoms were assessed. Questionnaires about personal and occupational risk factors were completed and suspicious cases were referred for NCV (nerve conduction velocity) testing and documentation of diagnosis. RESULTS: About 395 workers from automobile industry factories in Iran were assessed by interview and electrodiagnostic studies. Among 395 workers, 47 met the definition of CTS to yield a prevalence of 11.9%. These 47 workers averaged 29.85 years of age (SD = 6.28), and the mean age of the healthy group was 27.95 (SD = 4.86). 395 workers included 91 women (23%) and 304 men (77%). Using multivariate logistic regression model the largest adjusted odds ratios of personal and occupational factors for CTS were: exertion of force over one kilogram 6.38 (1.91-2.02); bending/twisting of the hands/wrists > 30 degrees , 5.62 (0.56-55.6); history of cigarette smoking 4.68 (1.80-11.80); rapid movement of hands 4.44 (1.41-14.02); and use of vibrating tools 3.23 (1.46-7.15). CONCLUSION: Some occupational factors including force exertion, bending/twisting of the hands, rapid movement of the hands and vibration are associated with CTS. PMID- 18418701 TI - The role of NRG3 in mammary development. AB - The Neuregulin gene family encodes EGF-containing ligands which mediate their effects by binding to the ERBB receptor tyrosine kinases, a signalling network with important roles in both mammary gland development and breast cancer. Neuregulin3 (NRG3), a ligand for ERBB4, promotes early mammary morphogenesis and acts during specification of the mammary placode, an aggregate of epithelial cells that forms during mid-embryogenesis. Recent studies have shown that NRG3 can alter the cell fate of other epidermal progenitor populations when NRG3 is mis-expressed throughout the basal layer of the developing epidermis with the K14 promoter. Here evidence for a key function for NRG3 in promoting early mammary morphogenesis and the implication for the role of NRG3 in breast cancer and establishment of the mammary lineage are discussed. PMID- 18418703 TI - Cysteamine prevents inhibition of thiol-containing enzymes caused by cystine or cystine dimethylester loading in rat brain cortex. AB - Cystinosis is a systemic genetic disease caused by a lysosomal transport deficiency accumulating cystine in the lysosomes of all tissues. Although tissue damage might depend on cystine accumulation, the mechanisms of tissue damage are still obscures. Considering that thiol-containing enzymes are critical for several metabolic pathways, our main objective was to investigate the effects of cystine or cystine dimethylester load on the thiol-containing enzymes creatine kinase and pyruvate kinase, in the brain cortex of young Wistar rats. The animals were injected twice a day with 1.6 micromol/g body weight of cystine dimethylester or 1 micromol/g body weight of cystine and/or 0.46 micromol/g body weight of cysteamine from the 16th to the 20th postpartum day and sacrificed after 12 h. Cystine or cystine dimethylester administration inhibited the two enzyme activities. Co-administration of cysteamine, the drug used to treat cystinotic patients, normalized the two enzyme activities. Lactate dehydrogenase activity, a nonthiol-containing enzyme was not affected by cystine dimethylester administration. Cystine inhibits creatine kinase and pyruvate activities possibly by oxidation of the sulfhydryl groups of the enzymes. Considering that creatine kinase and pyruvate kinase, like other thiol-containing enzymes, are crucial for energy homeostasis and antioxidant defenses, the enzymes inhibition caused by cystine released from lysosomes could be one of the mechanisms of tissue damage in patients with cystinosis. PMID- 18418704 TI - Pulmonary vein isolation using segmental versus electroanatomical circumferential ablation for paroxysmal atrial fibrillation: over 3-year results of a prospective randomized study. AB - PURPOSE: We tested the hypothesis that electroanatomic pulmonary vein (PV) antra encircling for the PV isolation will improve the outcome in treatment of paroxysmal atrial fibrillation (PAF), compared with segmental PV isolation. METHODS: Fifty-four patients underwent segmental PV isolation (group 1) and 56 patients circumferential PV isolation (group 2) for symptomatic PAF in a randomized study. RESULTS: Following single ablation procedure, at the 48 +/- 8 month follow-up, 30 (56%) and 32 (57%) patients in groups 1 and 2 remained free of arrhythmia (P = 0.41). After repeat ablation, 43 (80%) and 45 (80%) patients in groups 1 and 2 were free of arrhythmia without antiarrhythmic drugs (AADs); 48 (89%) and 51 (91%) patients in groups 1 and 2 did not have arrhythmia recurrences without or with AADs. CONCLUSION: This study demonstrates no advantage in long term arrhythmia-free clinical outcome after circumferential PV isolation in patients with frequent PAF. PMID- 18418705 TI - ACE-inhibitors, beta-blockers or the combination in heart failure: is it just an A-B-C ? : editorial to: effects of beta-blockade and ACE inhibition on B-type natriuretic peptides in stable patients with systolic heart failure by Rosenberg et al. PMID- 18418706 TI - Genomic sequence analysis of a granulovirus isolated from the Old World bollworm, Helicoverpa armigera. AB - The genome of a granulovirus isolated from the Old World bollworm, Helicoverpa armigera, was completely sequenced. The size of the Helicoverpa armigera granulovirus (HearGV) genome is 169,794 nt containing 179 open reading frames (ORFs), making it the second largest baculovirus genome analyzed to date. The genomes of HearGV and the Xestia c-nigrum GV (XecnGV) exhibit extensive sequence similarity and co-linearity, with both genomes containing the same nine homologous regions (hrs) with conserved structure and locations and sharing 167 open reading frames (ORFs). Phylogenetic inference and pairwise analysis of Kimura-2-parameter nucleotide distances for the lef-8, lef-9, and granulin genes indicate that HearGV is part of a cluster of granuloviruses typified by XecnGV. The HearGV genome contains all 62 ORFs found in common among other fully sequenced lepidopteran baculovirus genomes, as well as seven ORFs unique to HearGV. In addition, HearGV and XecnGV genomes share 20 ORFs not found among other baculovirus genomes sequenced to date. In addition to possessing ten ORFs with sequence similarity to baculovirus repeated ORFs (bro), the HearGV genome contains members of two other gene families with homologues in ascovirus, nucleopolyhedrovirus, and entomopoxvirus genomes. Alignment of the HearGV and XecnGV genome sequences revealed that HearGV is missing approximately 16.6 kbp of XecnGV-homologous sequence and contains approximately 8.2 kbp of sequence not found in the XecnGV genome. PMID- 18418709 TI - Detection and downregulation of type I IGF receptor expression by antibody conjugated quantum dots in breast cancer cells. AB - The type I insulin-like growth factor (IGF) receptor (IGF1R) is a transmembrane tyrosine kinase involved in breast cancer proliferation, survival, and metastasis. Several monoclonal antibodies directed against the receptor are in clinical trials. In order to develop a methodology to detect and measure IGF1R levels in breast cancer cells, we covalently conjugated an IGF1R antibody, AVE 1642, with quantum dots (Qdots), which are nanocrystals that emit fluorescence upon excitation. AVE-1642 Qdots only bound to cells that express IGF1R, and measured IGF1R levels by fluorescence emission at 655 nm. After binding to the cell surface, AVE-1642 Qdots underwent receptor mediated endocytosis, localized to endosome, and later translocated into the nucleus. Treating MCF-7 cells with AVE-1642 Qdots, but not unconjugated Qdots alone, downregulated IGF1R levels and rendered cells refractory to IGF-I stimulation. Furthermore, cell proliferation was slightly inhibited by AVE-1642 Qdots, but not the unconjugated Qdots. Our data suggest that AVE-1642 Qdots can be used to detect IGF1R expression and measure changes in cell surface receptor levels. In addition, the inhibitory effect of AVE-1642 Qdots to cell proliferation implies that it may serve as a traceable therapeutic agent. PMID- 18418707 TI - Sexual self schema as a moderator of sexual and psychological outcomes for gynecologic cancer survivors. AB - Gynecologic cancer patients are at high risk for emotional distress and sexual dysfunction. The present study tested sexual self schema as an individual difference variable that might be useful in identifying those at risk for unfavorable outcomes. First, we tested schema as a predictor of sexual outcomes, including body change stress. Second, we examined schema as a contributor to broader quality of life outcomes, specifically as a moderator of the relationship between sexual satisfaction and psychological statue (depressive symptoms and quality of life). A cross-sectional design was used. Gynecologic cancer survivors (N = 175) 2-10 years post treatment were assessed during routine follow up. In regression analyses controlling for sociodemographic variables, patients' physical symptoms/signs as evaluated by nurses, health status, and extent of partner sexual difficulties, sexual self schema accounted for significant variance in the prediction of current sexual behavior, responsiveness, and satisfaction. Moreover, schema moderated the relationship between sexual satisfaction and psychological outcomes, suggesting that a positive sexual self schema might "buffer" patients from depressive symptoms when their sexual satisfaction is low. Furthermore, the combination of a negative sexual self schema and low sexual satisfaction might heighten survivors' risk for psychological distress, including depressive symptomatology. These data support the consideration of sexual self schema as a predictor of sexual morbidity among gynecologic cancer survivors. PMID- 18418710 TI - Micro-flow visualization of red blood cell-enhanced platelet concentration at sudden expansion. AB - Microscopic steps and crevices are inevitable features within prosthetic blood contacting devices. This study aimed to elucidate the thrombogenicity of the associated microscopic flow features by studying the transport of fluorescent platelet-sized particles in a suspension of red blood cells (RBCs) flowing through a 100 microm:200 microm sudden expansion. Micro-flow visualization revealed a strong influence of hematocrit upon the path of RBCs and spatial concentration of particles. At all flow rates studied (Re = 8.3-41.7) and hematocrit 20% and lower, RBC streamlines were found to detach from the microchannel wall creating an RBC-depleted zone inside the step that was much larger than the cells themselves. However, the observed distribution of particles was relatively homogeneous. By contrast, the RBC streamlines of samples with hematocrit equal to or greater than 30% more closely followed the contour of the microchannel, yet exhibited enhanced concentration of particles within the corner. The corresponding size of the cell depletion layer was comparable with the size of the cells. This study implies that local platelet concentration in blood within the physiological range of hematocrit can be elevated within the flow separation region of a sudden expansion and implicates the role of RBCs in causing this effect. PMID- 18418711 TI - Inter- and intra-observer variability in detection and progression assessment with MRI of microadenoma in Cushing's disease patients followed up after bilateral adrenalectomy. AB - OBJECTIVE: To assess the inter- and intra-observer variability in detection and progression assessment with MRI of pituitary microadenomas in cases of Cushing's disease treated by adrenalectomy. DESIGN AND METHODS: Two radiologists independently reviewed initial and follow-up MRI studies of 32 patients with a history of Cushing's disease on two occasions each. Five classical qualitative signs and three quantitative measurements of microadenoma were considered. Intra and inter-observer reproducibility was evaluated with kappa statistics (qualitative signs) and intraclass correlation coefficients and Bland-Altman plots (quantitative measurements). RESULTS: No sign or measurement could be identified as better than others and none displayed consistently high levels of inter- or even intra-observer reproducibility (kappa or intraclass correlation coefficient > 0.80). Both qualitative and quantitative criteria showed lower reproducibility when used for diagnosing progression than when used for diagnosing the presence of an adenoma. Reproducibility of qualitative signs appeared to be influenced by reader's experience. CONCLUSIONS: Pituitary MRI study of patients with Cushing disease treated by adrenalectomy remains difficult. In practice, interpretation of MRI studies by a well-experienced reader should be preferred, unless a final decision can be reached by consensus between two and several readers. PMID- 18418712 TI - Dynamic tests for the diagnosis and assessment of treatment efficacy in acromegaly. AB - In the vast majority of cases, basal serum GH and IGF-1 levels are markedly increased in patients with obvious clinical signs and symptoms of acromegaly. The oral glucose tolerance test (OGTT) is useful for diagnosis in the minority of patients who have weak GH hypersecretion. The cutoff for a "normal" GH nadir in the OGTT remains to be agreed. The type of GH assay, its sensitivity, the type of standard used by the manufacturer, the patient's age and especially gender, must all be taken into account. Recent studies using new highly sensitive assays suggest an upper normal GH nadir of 0.71 microg/l for female healthy patients, but no "universal" cut-off has yet been defined for healthy males (from 0.057 to 0.25 microg/l). The 1 microg/l cutoff proposed for the diagnosis of acromegaly in a 2000 consensus should be abandoned in favor of a 0.30 microg/l cutoff. Clinicians should know which assay is used, together with its sensitivity and the standard, before making therapeutic decisions. A more pragmatic view should probably be adopted when assessing the treatment response. Indeed, if "cure" is defined not with the <1 microg/l GH nadir but on the basis of healthy control values, many patients will not be considered controlled. However, the clinical relevance of such goal (e.g. achieving GH nadir <0.4 microg/l rather than <1 microg/l) in terms of prognosis and prediction of outcome on long term is not firmly established. Thus, from a pragmatic point of view, achieving a normal age adjusted IGF-1 level and a GH nadir below 1 microg/l during OGTT will probably remain relevant for defining remission and good disease control in terms of morbidity and mortality in acromegaly. PMID- 18418713 TI - Secondary hypoadrenalism. AB - Secondary adrenal insufficiency (SAI) is a clinical disorder that results from hypothalamic or hypophyseal damage or from prolonged administration of supraphysiological doses of glucocorticoids. Since glucocorticoids are widely used for a variety of diseases, the prevalence of SAI is by far exceeding that of primary adrenal insufficiency. Although the presentation of adrenal insufficiency may be insidious and difficult to recognize, an appropriate adrenocortical hormone replacement could lead to a normal quality of life and longevity can be achieved. The spectrum of adrenal insufficiency ranges from overt adrenal crises to subtle dysfunctions in asymptomatic patients who may be at risk of developing acute adrenal insufficiency since their hypothalamic-pituitary-adrenal (HPA) axis cannot appropriately react to stress. Thus, identification of patients with subtle abnormalities of the HPA is mandatory for avoiding this life-threatening event in stressful conditions. The optimal tests and the optimal testing sequence for adrenal insufficiency are still matter of debate. Insulin tolerance test (ITT) could be the gold standard, as it tests the whole HPA axis, but there are some patients who pass the ITT failing the ACTH test. Various alternatives to the ITT, including the standard cosyntropin stimulation test (SST) and low-dose SST, have been proposed since the adrenal gland in SAI loses the capacity for a prompt response to ACTH stimulation. The standard ACTH dose, but not the 1 microg dose, increases adrenal blood flow and this may contribute to produce an early cortisol response of greater magnitude. Moreover, the loss of the early cortisol response to ACTH stimulation could be a specific property of adrenal insufficiency, thus being a sensitive and early marker of failing adrenal function. While the results of the SSTs are often positive in patients with long-standing and severe disease, in patients with mild or recent-onset SAI these tests, using either 250 microg or 1 microg ACTH, tend to give normal results; thus, a negative cosyntropin test result does not rule out the possibility of SAI. Further studies with a systematic comparison of the different tests in large series of patients submitted to a prolonged follow-up are needed to solve the controversy of the optimal diagnostic strategy of SAI. PMID- 18418714 TI - Recidivism in HIV-infected incarcerated adults: influence of the lack of a high school education. AB - Recidivism is a pervasive problem facing the incarcerated. Incarcerated persons who are human immunodeficiency virus (HIV)-infected often have multiple risk factors associated with initial incarceration and recidivism, in particular, injection drug use. Yet, some jails provide case management for HIV-infected inmates to provide continuity of health care, which might have positive effects on reentry into the community. We sought to measure recidivism and factors related to recidivism in an HIV-infected cohort in an urban county jail with an active case management program. Fifty-two inmates surveyed in 1999 at the San Francisco County Jail were followed for rearrests through 2006. In follow-up, 73% were re-incarcerated on an average of 6.8 times for 552 days. Risk factors included nonwhite ethnicity, history of homelessness and crack use, common risk factors for incarceration. Less than high school education was associated with recidivism, shorter time to reincarceration, and more incarcerations. HIV infected inmates spend a high proportion of time in multiple incarcerations, a reflection of the cyclical nature of incarceration despite comprehensive case management. Well-known risk factors for incarceration were associated with recidivism; in addition, lack of high school education played a prominent role. Education should be explored as a way to make further progress on breaking the cycle of incarceration. PMID- 18418715 TI - A new tool for probing of cell-cell communication: human embryonic germ cells inducing apoptosis of SKOV3 ovarian cancer cells on a microfluidic chip. AB - A microfluidic device with unidirectional perfusion has been developed to observe the effect of human embryonic germ (hEG) cells on SKOV3 cells. The hEG and SKOV3 cells were seeded in the inlet and the outlet reservoirs separately, and co cultured for 2 days. The medium was perfused unidirectionally from the inlet to the outlet. The growth inhibition of SKOV3 cells was monitored online and the apoptosis signals in SKOV3 culture area decreased along the flow of the medium. In conclusion, microfluidic chip is a potentially useful tool to investigate the effect of stem cells on cancer cells with intuitionistic cell-based screens. PMID- 18418716 TI - Mouse monoclonal antibodies in biological research: strategies for high throughput production. AB - Mouse monoclonal antibodies have become key components in basic research as well as in the clinical laboratory. Being invaluable tools in many biological assays, they continue to be the primary choice in the research field, although the conventional technology used for hybridoma generation and screening is a still lengthy, time-consuming and low-throughput process. With the advent of genetic immunisation and the application of automation and microarray to the traditional biological assays, the monoclonal antibody field has been revolutionised. Here, we will briefly review the most relevant strategies which have made the manufacture of murine monoclonal antibodies a faster and high-throughput technology. PMID- 18418717 TI - Low migrant mortality in Germany for men aged 65 and older: fact or artifact? AB - Migrant mortality in Europe was found to be lower than mortality of host populations. In Germany, residents with migrant background constitute nearly one tenth of the population aged 65+ with about 40% of them being foreigners. The German Pension Scheme follows vital status of pensioners very accurately. Mortality re-estimation reveals two-fold underestimation of mortality of foreigners due to biased death numerator and population denominator. PMID- 18418718 TI - Organization of unusual idiosomal glands in a water mite, Teutonia cometes (Teutoniidae). AB - The unusual idiosomal glands of a water mite Teutonia cometes (Koch 1837) were examined by means of transmission and scanning electron microscopy as well as on semi-thin sections. One pair of these glands is situated ventrally in the body cavity of the idiosoma. They run posteriorly from the terminal opening (distal end) on epimeres IV and gradually dilate to their proximal blind end. The terminal opening of each gland is armed with the two fine hair-like mechanoreceptive sensilla ('pre-anal external' setae). The proximal part of the glands is formed of columnar secretory epithelium with a voluminous central lumen containing a large single 'globule' of electron-dense secretory material. The secretory gland cells contain large nuclei and intensively developed rough endoplasmic reticulum. Secretory granules of Golgi origin are scattered throughout the cell volume in small groups and are discharged from the cells into the lumen between the scarce apical microvilli. The distal part of the glands is formed of another cell type that is not secretory. These cells are composed of narrow strips of the cytoplasm leaving the large intracellular vacuoles. A short excretory cuticular duct formed by special excretory duct cells connects the glands with the external medium. At the base of the terminal opening a cuticular funnel strengthens the gland termination. At the apex of this funnel a valve prevents back-flow of the extruded secretion. These glands, as other dermal glands of water mites, are thought to play a protective role and react to external stimuli with the help of the hair-like sensilla. PMID- 18418719 TI - Non-invasive cardiac output by transthoracic electrical bioimpedence in post cardiac surgery patients: comparison with thermodilution method. AB - OBJECTIVE: Thoracic electrical bioimpedance (TEB) cardiac output (CO) is being explored increasingly as a non-invasive alternative to the pulmonary artery catheter (PAC). This study compared TEB-CO measured using a new instrument- NICOMON (Larsen & Toubro Ltd. India) with thermodilution (Td) CO in post-cardiac surgery patients. METHODS: Postoperative cardiac surgical patients requiring a PAC for their management were studied. TEB-CO was measured by passing a 4 mA RMS alternating current across the chest and measuring the analog bioimpedence across the thorax. Kubicek equation was used to estimate TEB-CO. Td-CO was measured using a PAC. Bland-Altman analysis was used to compare paired data. RESULTS: One hundred and ninety-seven pairs of CO measurements were made by the two methods among 35 patients. Mean TEB-CO was 5.15 +/- 1.27 l/min and mean Td-CO was 5.22 +/ 1.28 l/min. Pearson correlation coefficient (r) for these measurements was 0.856 (P < 0.01), with bias -0.0651 l and precision: +/-1.37 l/min. The percentage error of measurement of this precision was 26.44%. Cardiac index also correlated among the two methods (r = 0.789; P = 0.01). CONCLUSIONS: Thoracic electrical bioimpedance cardiac output compares favorably with thermodilution method among post-cardiac surgery patients. Further studies are indi- cated with this instrument to validate its efficacy in various clinical situations and utility in monitoring hemodynamic interventions. PMID- 18418720 TI - Assessment of heavy metals contamination and leaching characteristics in highway side soils, Iran. AB - Twenty seven topsoil samples were collected under stable weather conditions from the northern and southern sides of Tehran-Karaj Highway, Iran. Samples were analyzed for heavy metal (Pb, V, Zn, Ni, Co, Cr, Cd, and Mn) contents and leaching characteristics. The results showed that all heavy metal contents except Cr, Mn and Co are higher than acceptable values in natural soils. High contents of these elements could be attributed to anthropogenic effects related to traffic sources. Toxicity characteristics leaching procedure (TCLP) test results reveal that the studied soils may be hazardous. Statistical analysis shows significant positive correlations between heavy metals and organic matter. Also a significant correlation was observed between Cd, Pb and Zn. This affinity and exponentially decreasing concentration with distance from the edge of the road suggest that automobiles are a major source of these metals in the roadside soils. Use of leaded gasoline and tire erosion gives a boost to this claim. PMID- 18418721 TI - Study of chemical composition and morphology of airborne particles in Chandigarh, India using EDXRF and SEM techniques. AB - The elemental composition and morphology of aerosols, collected from March 95 to February 96 and March 96 to August 96 respectively in the city of Chandigarh, India is determined using Energy Dispersive X-ray fluorescence and scanning electron microscopic techniques. The elemental concentration levels are found to be higher by a factor of 2-7 in the spring season as compared to the rainy season. The concentration of spherical and non-spherical (i.e. elongated) aerosols is more in the spring season and is reduced drastically in the rainy season due to the prominent wash out effect of rains. More accurate particle classification and source identification is obtained when based on combination of chemical composition and particle morphology. Possible sources identified from this analysis are soil dust, Industrial activity, Agricultural and Garbage burning, Maritime aerosols and Automobile exhaust. PMID- 18418722 TI - Development of a data management framework in support of southeastern tidal creek research. AB - The NOAA Center of Excellence for Oceans and Human Health Initiative (OHHI) at the Hollings Marine Laboratory (HML) is developing a data management framework that supports an integrated research program across scientific disciplines. The primary focus of the database is to support environmental research focused on tidal creek watershed systems. Specifically, the current data holdings include physical water quality parameters, nutrients, pathogens, chemical contaminants, benthic and nekton species abundances and human dimensions data from Georgia, North Carolina and South Carolina dating to 1994. These data are not from a single long-term research project but are derived from several state and federal research programs and integrated into a common database model to support current research being conducted under the OHHI program at HML. The Tidal Creek database was developed with the intent to support a well documented and open system, thus metadata elements from common metadata standards including the Dublin Core ISO 15836:2003 and Federal Geographic Data Committee (FGDC-STD-001-1998) are components of the database model. The result is a semantic database framework with descriptive ancillary data at the record level including methods, investigator names, date, locations and other descriptive elements. The primary users of the database are project personnel to meet analytical needs. The database is also available through a number of web-based applications that are designed to give users the necessary information to evaluate and access data. In addition, data can be accessed with Open Geospatial Consortium (OGC) standards, and species records and abundances are being made available to the Ocean Biogeographic Information System (OBIS). Overall, the Tidal Creek database summarizes the response of tidal creeks and watersheds to coastal development, and serves as a repository for environmental, demographic, and socioeconomic data in the Southeast. PMID- 18418723 TI - Are deep strategic learners better suited to PBL? A preliminary study. AB - The aim of this study was to determine if medical students categorized as having deep and strategic approaches to their learning find problem-based learning (PBL) enjoyable and supportive of their learning, and achieve well in the first-year course. Quantitative and qualitative data were gathered from first-year medical students (N = 213). All students completed the Medical Course Learning Questionnaire at the commencement and completion of their first year of medical studies. The instrument measured a number of different aspects of learning, including approaches to learning, preferences for different learning environments, self-efficacy, and perceptions of learning within PBL tutorials. Qualitative data were collected from written responses to open questions. Results of students' performance on two forms of examinations were obtained for those giving permission (N = 68). Two-step cluster analysis of the cohort's responses to questions about their learning approaches identified five clusters, three of which represented coherent combinations of learning approaches (deep, deep and strategic, and surface apathetic) and two clusters which had unusual or dissonant combinations. Deep, strategic learners represented 25.8% of the cohort. They were more efficacious, preferred learning environments which support development of understanding and achieved significantly higher scores on the written examination. Strongly positive comments about learning in PBL tutorials were principally described by members of this cluster. This preliminary study employed a technique to categorize a student cohort into subgroups on the basis of their approaches to learning. One, the deep and strategic learners, appeared to be less vulnerable to the stresses of PBS in a medical course. While variation between individual learners will always be considerable, this analysis has enabled classification of a student group that may be less likely to find PBL problematic. Implications for practice and suggestions for future research are proposed. PMID- 18418724 TI - Impact of curriculum on understanding of professional practice: a longitudinal study of students commencing dental education. AB - This longitudinal study examines changes in understanding of dental practice among a cohort of students in the early years of a dentistry programme. In their first two professional years, we identified five distinct understandings of dental practice that we have ordered from least to most comprehensive: relieving pain or generally caring for teeth, carrying out particular dental procedures, diagnosing and treating dental problems or diseases, evaluating and responding to oral health, and finally, evaluating oral health and preventing oral disease in the community. At entry into the dental program the most common understandings among both men and women focused on dental procedures or diagnosis and treatment. The largest changes in students' responses at the end of the first and second professional years were generally in line with the emphasis of the curriculum in each of these 2 years, although prevention was not clearly featured. These data suggest that at least some students responded to the curriculum and, hence, highlight the impact of the curriculum on students' emerging understandings. We conclude that curricula can have a key role in the development of understanding of professional practice during professional programmes, although the impact of curricula is not always as expected and merits investigation. PMID- 18418725 TI - Modeling manipulation in medical education. AB - As residents and medical students progress through their medical training, they are presented with multiple instances in which they feel they must manipulate the healthcare system and deceive others in order to efficiently treat their patients. This, however, creates a culture of manipulation resulting in untoward effects on trainees' ethical and professional development. Yet manipulation need not be a skill necessary to practice medicine, and steps should be taken by both individuals and institutions to combat the view that the way medicine must be practiced "in the real world" is somehow different from what one's affective moral sense implores. PMID- 18418726 TI - Immunohistochemical staining for tilapia and human insulin demonstrates that a tilapia transgenic for humanized insulin is a mosaic. PMID- 18418727 TI - Nephrogenic systemic fibrosis: an emerging entity. AB - Nephrogenic systemic fibrosis (NSF) is a progressive disorder which has been seen only in patients with chronic kidney disease. It is associated with fibrosis of the skin and connective tissue throughout the body. The skin thickens and becomes hard, rigid, and coarse, which severely restricts movement of the joints. It can also lead to widespread fibrosis of internal organs involving lungs, heart, diaphragm, esophagus, and skeletal muscle, and in some cases may lead to death. Based on case series reports and skin biopsy showing gadolinium (Gd) ions, NSF has been directly linked to Gd contrast exposure given during MRI/MRA. There are over 250 reported cases of NSF worldwide with hundreds still not reported or remaining undiagnosed. Symptoms of NSF appear within 2-75 days, with a mean of 25 days after exposure to Gd contrast. This disorder is not well understood, and more research is needed to obtain information about how Gd causes this condition. Currently there is no effective treatment, so prevention is the only way to avoid this serious illness. PMID- 18418728 TI - Factors influencing blood pressure control in children and adolescents. AB - OBJECTIVE: To investigate the incidence of increased blood pressure among children and adolescents and its relation with the anthropometric indexes, fasting insulin, insulin sensitivity indexes and lipid levels. BACKGROUND: Sufficient evidence suggests that the incidence of essential hypertension among children and adolescents is rapidly increased parallel to the incidence of obesity among these subjects. METHODS: We investigated 154 school-aged children; 77 males and 77 females, aged 11.28+/-2.99 (4-18) years old. After an overnight (12 h) fasting, we measured the following: anthropometric parameters, insulin (micro units/ml), glucose (mg/dl), cholesterol (mg/dl), cholesterol-HDL (mg/dl), cholesterol-LDL (mg/dl), triglycerides (mg/dl), uric acid (mg/dl), systolic and diastolic blood pressure (BP) (mmHg), HOMA-R index, QUICKI index and estimated glomerular filtration rate (eGFR). RESULTS: Boys showed greater body weight than girls as well as greater height and waist circumference. High normal systolic and diastolic BP measurements were more frequent among boys (HN-SBP: 27.2% vs. 16.8%, P=0.002. HN-DBP: 23.3% vs. 16.8%, P=0.05), but girls showed a higher frequency of systolic and diastolic high BP (H-SBP: 5.2% vs. 3.8%, P=0.6 (NS). H-DBP: 10.4% vs. 3.8%, P=0.05). Girls showed higher insulin levels and HOMA-R index values than boys, as well as higher levels of total cholesterol, triglycerides and LDL cholesterol. Boys exhibited greater values of estimated glomerular filtration rate than girls. The most important factors influencing systolic and diastolic BP were the waist circumference (central obesity) and the body weight. Fasting insulin levels showed a biphasic effect upon systolic and, especially, diastolic BP. Normoinsulinemia exhibited a positive effect upon systolic and diastolic BP but high insulin levels exhibited a negative effect upon diastolic BP. No considerable effect of lipid parameters was found upon BP. CONCLUSIONS: Our results suggest that anthropometric indexes, particularly central obesity and body weight play the most important role in BP control among children and adolescents. Girls exhibit greater frequency of high BP than boys, as well as greater frequency of other metabolic syndrome risk factors. Insulin levels and insulin sensitivity are important factors influencing BP control. Insulin, per se, plays a pivotal role in BP control, normoinsulinemia produce a positive effect upon BP, but high insulin levels exert a hypotensive action. PMID- 18418729 TI - Friends and foes: streptomycetes as modulators of plant disease and symbiosis. AB - The ecological role of soil streptomycetes within the plant root environment is currently gaining increased attention. This review describes our recent advances in elucidating the complex interactions between streptomycetes, plants, pathogenic and symbiotic microorganisms. Streptomycetes play diverse roles in plant-associated microbial communities. Some act as biocontrol agents, inhibiting plant interactions with pathogenic organisms. Owing to the antagonistic properties of streptomycetes, they exert a selective pressure on soil microbes, which may not always be for plant benefit. Others promote the formation of symbioses between plant roots and microbes, and this is in part due to their direct positive influence on the symbiotic partner, expressed as, e.g., promotion of hyphal elongation of symbiotic fungi. Recently, streptomycetes have been identified as modulators of plant defence. By repressing plant responses to pathogens they facilitate root colonisation with pathogenic fungi. In contrast, other strains induce local and systemic resistance against pathogens or enhance plant growth. In conclusion, while streptomycetes have a clear potential of acting as biocontrol agents, care has to be taken to avoid strains that select for virulent pathogens or enhance disease development. We argue towards the use of an integrated screening approach in the search for efficient biocontrol agents, including assays on in vitro antagonism, plant growth, and disease suppression. PMID- 18418730 TI - Growth inhibition induced by transforming growth factor-beta1 in human oral squamous cell carcinoma. AB - Oral squamous cell carcinoma (OSCC) is a world-wide health problem and its incidence accounts for 1.9-3.5% of all malignant tumors. Transforming growth factor beta/Smads (TGF-beta/Smads) signaling pathway plays an important role in oncogenesis, but its function and molecular mechanisms in OSCC remain unclear. Expression of transforming growth factor-beta receptor type II (TbetaRII) and Smad4 was studied by immunohistochemistry in 108 OSCC patients and 10 normal controls. Function and molecular mechanisms of TGF-beta/Smads signaling pathway was then investigated in two human tongue squamous carcinoma cell lines with high and low metastasis (Tb and Tca8113) by RT-PCR, Western Blot, immunofluorescence, cell growth curve and flow cytometry (FCM), respectively. TbetaRII and Smad4 were significantly down-regulated in tumor tissues (with or without lymph node metastasis) compared to normal oral epithelium tissues (P < 0.05). TGF-beta1 induced arrest of the cell cycle rather than cell death in Tca8113 and Tb cells, and this influence was mediated by the increasing the expression and changing the location of its downstream components of TGF-beta1/Smads signaling pathway. TGF beta1 rapidly increased the expression of p15 and p21 in both Tca8113 and Tb cells. TGF-beta1 did not increase p27 expression in Tca8113 cells, but p27 expression was increased in Tb cells. These indicated that TGF-beta1 induced G(1) arrest of cell cycle through a different regulating pathway in Tb cells compared with Tca8113 cells. Thus, we conclude that TGF-beta/Smads signaling pathway play a important role on cell growth and metastasis potential in OSCC. PMID- 18418731 TI - Heat shock proteins HSP70 and HSP27 in the cerebral spinal fluid of patients undergoing thoracic aneurysm repair correlate with the probability of postoperative paralysis. AB - An understanding of the time course and correlation with injury of heat shock proteins (HSPs) released during brain and/or spinal cord cellular stress (ischemia) is critical in understanding the role of the HSPs in cellular survival, and may provide a clinically useful biomarker of severe cellular stress. We have analyzed the levels of HSPs in the cerebrospinal fluid (CSF) from patients who are undergoing thoracic aneurysm repair. Blood and CSF samples were collected at regular intervals, and CSF was analyzed by enzyme-linked immunosorbent assay for HSP70 and HSP27. These results were correlated with intraoperative somatosensory-evoked potentials measurements and postoperative paralysis. We find that the levels of these proteins in many patients are elevated and that the degree of elevation correlates with the risk of permanent paralysis. We hypothesize that sequential measurement intraoperatively of the levels of the heat shock proteins HSP70 and HSP27 in the CSF can predict those patients who are at greatest risk for paralysis during thoracic aneurysm surgery and will allow us to develop means of preventing or attenuating this severe and often fatal complication. PMID- 18418732 TI - Protein kinase C signaling during T cell activation induces the endoplasmic reticulum stress response. AB - T cell receptor (TCR) ligation (signal one) in the presence of co-stimulation (signal two) results in downstream signals that increase protein production enabling naive T cells to fully activate and gain effector function. Enhanced production of proteins by a cell requires an increase in endoplasmic reticulum (ER) chaperone expression, which is accomplished through activation of a cellular mechanism known as the ER stress response. The ER stress response is initiated during the cascade of events that occur for the activation of many cells; however, this process has not been comprehensively studied for T cell function. In this study, we used primary T cells and mice circulating TCR transgenic CD8(+) T cells to investigate ER chaperone expression in which TCR signaling was initiated in the presence or absence of co-stimulation. In the presence of both signals, in vitro and in vivo analyses demonstrated induction of the ER stress response, as evidenced by elevated expression of GRP78 and other ER chaperones. Unexpectedly, ER chaperones were also increased in T cells exposed only to signal one, a treatment known to cause T cells to enter the 'nonresponsive' states of anergy and tolerance. Treatment of T cells with an inhibitor to protein kinase C (PKC), a serine/threonine protein kinase found downstream of TCR signaling, indicated PKC is involved in the induction of the ER stress response during the T cell activation process, thus revealing a previously unknown role for this signaling protein in T cells. Collectively, these data suggest that induction of the ER stress response through PKC signaling is an important component for the preparation of a T cell response to antigen. PMID- 18418733 TI - Rapamycin conditionally inhibits Hsp90 but not Hsp70 mRNA translation in Drosophila: implications for the mechanisms of Hsp mRNA translation. AB - Rapamycin inhibits the activity of the target of rapamycin (TOR)-dependent signaling pathway, which has been characterized as one dedicated to translational regulation through modulating cap-dependent translation, involving eIF4E binding protein (eIF4E-BP) or 4E-BP. Results show that rapamycin strongly inhibits global translation in Drosophila cells. However, Hsp70 mRNA translation is virtually unaffected by rapamycin treatment, whereas Hsp90 mRNA translation is strongly inhibited, at normal growth temperature. Intriguingly, during heat shock Hsp90 mRNA becomes significantly less sensitive to rapamycin-mediated inhibition, suggesting the pathway for Hsp90 mRNA translation is altered during heat shock. Reporter mRNAs containing the Hsp90 or Hsp70 mRNAs' 5' untranslated region recapitulate these rapamycin-dependent translational characteristics, indicating this region regulates rapamycin-dependent translational sensitivity as well as heat shock preferential translation. Surprisingly, rapamycin-mediated inhibition of Hsp90 mRNA translation at normal growth temperature is not caused by 4E-BP mediated inhibition of cap-dependent translation. Indeed, no evidence for rapamycin-mediated impaired eIF4E function is observed. These results support the proposal that preferential translation of different Hsp mRNA utilizes distinct translation mechanisms, even within a single species. PMID- 18418734 TI - Adrenomedullin-2/intermedin induces cAMP accumulation in dissociated rat spinal cord cells: evidence for the existence of a distinct class of sites of action. AB - Adrenomedullin-2/intermedin is structurally related to the calcitonin family of peptides, which includes calcitonin gene-related peptide (CGRP), adrenomedullin, and amylin. We recently reported that CGRP and adrenomedullin act through distinct receptors to induce cyclic adenosine monophosphate (cAMP) accumulation in dispersed cells from embryonic rat spinal cord. Here, we investigated the apparent affinity and efficacy of adrenomedullin-2/intermedin for these receptors. Adrenomedullin-2/intermedin competed with [(125)I]-CGRP for binding to specific embryonic spinal cord cells with a pIC(50) of 9.73 +/- 0.06. Interestingly, adrenomedullin-2/intermedin competed for specific [(125)I] adrenomedullin binding in a biphasic manner with pIC(50) of 9.03 +/- 0.22 and 6.45 +/- 0.24, respectively. Cellular levels of cAMP were increased by adrenomedullin-2/intermedin (pEC(50) 7.84 +/- 0.08) when cells were exposed to this peptide for 10 min at 37 degrees C. This effect was partially inhibited by the non-peptide antagonist BIBN4096BS (pA(2) 6.56 +/- 0.12), the adrenomedullin antagonist hAM(22-52) (pA(2) 6.36 +/- 0.30), and the adrenomedullin/CGRP antagonist CGRP(8-37) (pA(2) 7.24 +/- 0.60). More interestingly, a highly significant effect of adrenomedullin-2/intermedin on cAMP accumulation (pEC(50) 7.3 +/- 0.14) was still observed even in the presence of a mixture of saturating concentrations of BIBN4096BS, hAM(22-52), and the amylin antagonist AC187. Taken together, these data provide evidence for the possible existence of a distinct class of receptor sites for adrenomedullin-2/intermedin in embryonic rat spinal cord cells. PMID- 18418735 TI - PACAP-mediated neuroprotection of neurochemically identified cell types in MSG induced retinal degeneration. AB - Pituitary adenylate cyclase-activating polypeptide (PACAP) is neuroprotective in animal models of different brain pathologies and injuries, including cerebral ischemia, Parkinson's disease, and different types of retinal degenerations. We have previously shown that PACAP is protective against monosodium glutamate (MSG) induced retinal degeneration, where PACAP-treated retinas has more retained structure and PACAP induces anti-apoptotic while it inhibits pro-apoptotic signaling pathways. The aim of the present study was to investigate cell-type specific effects of PACAP in MSG-induced retinal degeneration by means of immunohistochemistry. Rat pups received MSG (2 mg/g b.w.) applied on postnatal days 1, 5, and 9. PACAP (100 pmol in 5 microl saline) was injected into the right vitreous body, while the left eye received only saline. Retinas were processed for immunocytochemistry after 3 weeks. Immunolabeling was determined for vesicular glutamate transporter 1, tyrosine hydroxylase, calretinin, calbindin, parvalbumin, and vesicular gamma-aminobutyric acid (GABA) transporter. In the MSG treated retinas, the cell bodies and processes in the inner nuclear, inner plexiform, and ganglion cell layers displayed less immunoreactivity for all antisera. Apart from photoreceptors, only one major retinal cell type examined in this study; the calbindin-immunoreactive horizontal cell seemed not to be affected by MSG application. After simultaneous application of MSG and PACAP, staining of retinas was similar to that of normal eyes, with no significant alterations in immunoreactive patterns. These findings further support the neuroprotective function of PACAP in MSG-induced retinal degeneration. PMID- 18418737 TI - Bundled slash: a potential new biomass resource for fuels and chemicals. AB - Postharvest residues for southern pine species have not previously been quantified to compare volumes produced from both thinnings and clearcut volumes. A John Deere 1490 Slash Bundler bundled postharvest residues following a first thinning of a 14-year-old stand, a second thinning of a 25-year-old stand, and a clearcut of a naturally regenerated mature stand of 54 years of age. Regardless of stand type, nearly one-fifth of merchantable volume harvested was collected as postharvest residue. Initial bundle moisture contents were 127.3, 81.1, and 49.4% dry basis (db) for the first and second thinnings and mature stands, respectively. Bundle needles content was found to significantly influence the relative moisture contents of the bundles by stand type due to the high moisture content of needles compared to other bundle components. Bundles were stored outside and exposed to very hot and dry conditions and dried very rapidly to lowest moisture contents of 22.8, 14.5, and 13.5% (db) for first and second thinnings and mature stands, respectively. Response to moderating temperatures and higher precipitation resulted in rapid moisture content increase to 69.9, 46.2, and 38.1% (db) for the first and second thinnings and mature stand bundles by the end of the study. Temperature and precipitation and bundle percentage needles content all significantly influenced the rapid moisture content variations observed over the study periods. PMID- 18418736 TI - The evolutionary history and tissue mapping of amino acid transporters belonging to solute carrier families SLC32, SLC36, and SLC38. AB - Members of the solute carrier families (SLC) 32, 36, and 38, together also designated the beta-group of SLCs, are known to transport neutral amino acids. In this paper, we show that these three families were present before the split of the animal lineage and that they are likely to share a common decent. We also show that the APF transporters found in plants are most likely homologous to the mammalian beta-group, suggesting that this type of transporters arouse early in the evolution of eukaryotes. We performed detailed tissue expression analysis of all the members of the beta-group in rat and found several examples of highly specific expression patterns, with SLC38A7 being exclusively found in liver, SLC38A5 in blood, and SLC38A4 in muscle and liver. Moreover, we found that SLC38A10 is expressed in several endocrine organs. We also found that SLC38A1 is highly up regulated in the cortex from rats treated with diazepam and that SLC38A2 is significantly down regulated in the same tissue. In addition, we performed a detailed expression analysis of SLC38A1 and SLC38A6 in mouse brain using in situ hybridization, which showed that both these transporters are widely expressed in the brain. PMID- 18418738 TI - Pretreatment of whole-crop harvested, ensiled maize for ethanol production. AB - To have all-year-round available feedstock, whole-crop maize is harvested premature, when it still contains enough moisture for the anaerobic ensiling process. Silage preparation is a well-known procedure for preserving plant material. At first, this method was applied to obtain high-quality animal feed. However, it was found that such ensiled crops are very suitable for bioenergy production. Maize silage, which consists of hardly degradable lignocellulosic material, hemicellulosic material, and starch, was evaluated for its potential as a feedstock in the production of bioethanol. It was pretreated at low severity (185 degrees C, 15 min) giving very high glucan ( approximately 100%) and hemicellulose recoveries (<80%)-as well as very high ethanol yield in simultaneous saccharification and fermentation experiments (98% of the theoretical production based on available glucan in the medium). The theoretical ethanol production of maize silage pretreated at 185 degrees C for 15 min without oxygen or catalyst was 392 kg ethanol per ton of dry maize silage. PMID- 18418739 TI - Enzymatic hydrolysis and ethanol fermentation of high dry matter wet-exploded wheat straw at low enzyme loading. AB - Wheat straw was pretreated by wet explosion using three different oxidizing agents (H(2)O(2), O(2), and air). The effect of the pretreatment was evaluated based on glucose and xylose liberated during enzymatic hydrolysis. The results showed that pretreatment with the use of O(2) as oxidizing agent was the most efficient in enhancing overall convertibility of the raw material to sugars and minimizing generation of furfural as a by-product. For scale-up of the process, high dry matter (DM) concentrations of 15-20% will be necessary. However, high DM hydrolysis and fermentation are limited by high viscosity of the material, higher inhibition of the enzymes, and fermenting microorganism. The wet-explosion pretreatment method enabled relatively high yields from both enzymatic hydrolysis and simultaneous saccharification and fermentation (SSF) to be obtained when performed on unwashed slurry with 14% DM and a low enzyme loading of 10 FPU/g cellulose in an industrial acceptable time frame of 96 h. Cellulose and hemicellulose conversion from enzymatic hydrolysis were 70 and 68%, respectively, and an overall ethanol yield from SSF was 68%. PMID- 18418740 TI - Substrate dependency and effect of xylanase supplementation on enzymatic hydrolysis of ammonia-treated biomass. AB - Pretreatment based on aqueous ammonia was investigated under two different modes of operation: soaking in aqueous ammonia and ammonia recycle percolation. These processes were applied to three different feedstocks with varied composition: corn stover, high lignin (HL), and low lignin (LL) hybrid poplars. One of the important features of ammonia-based pretreatment is that most of the hemicellulose is retained after treatment, which simplifies the overall bioconversion process and enhances the conversion efficiency. The pretreatment processes were optimized for these feedstocks, taking carbohydrate retention as well as sugar yield in consideration. The data indicate that hybrid poplar is more difficult to treat than corn stover, thus, requires more severe conditions. On the other hand, hybrid poplar has a beneficial property that it retains most of the hemicellulose after pretreatment. To enhance the digestibility of ammonia treated poplars, xylanase was supplemented during enzymatic hydrolysis. Because of high retention of hemicellulose in treated hybrid poplar, xylanase supplementation significantly improved xylan as well as glucan digestibility. Of the three feedstocks, best results and highest improvement by xylanase addition was observed with LL hybrid poplar, showing 90% of overall sugar yield. PMID- 18418741 TI - Alkali (NaOH) pretreatment of switchgrass by radio frequency-based dielectric heating. AB - Radio-frequency (RF)-based dielectric heating was used in the alkali (NaOH) pretreatment of switchgrass to enhance its enzymatic digestibility. Due to the unique features of RF heating (i.e., volumetric heat transfer, deep heat penetration of the samples, etc.), switchgrass could be treated on a large scale, high solid content, and uniform temperature profile. At 20% solid content, RF assisted alkali pretreatment (at 0.1 g NaOH/g biomass loading and 90 degrees C) resulted in a higher xylose yield than the conventional heating pretreatment. The enzymatic hydrolysis of RF-treated solids led to a higher glucose yield than the corresponding value obtained from conventional heating treatment. When the solid content exceeded 25%, conventional heating could not handle this high-solid sample due to the loss of fluidity, poor mixing, and heating transfer of the samples. As a result, there was a significantly lower sugar yield, but the sugar yield of the RF-based pretreatment process was still maintained at high levels. Furthermore, the optimal particle size and alkali loading in the RF pretreatment was determined as 0.25-0.50 mm and 0.25 g NaOH/g biomass, respectively. At alkali loading of 0.20-0.25 g NaOH/g biomass, heating temperature of 90(o)C, and solid content of 20%, the glucose, xylose, and total sugar yield from the combined RF pretreatment and the enzymatic hydrolysis were 25.3, 21.2, and 46.5 g/g biomass, respectively. PMID- 18418742 TI - Biological hydrogen production using chloroform-treated methanogenic granules. AB - In fermentative hydrogen production, the low-hydrogen-producing bacteria retention rate limits the suspended growth reactor productivity because of the long hydraulic retention time (HRT) required to maintain adequate bacteria population. Traditional bacteria immobilization methods such as calcium alginate entrapment have many application limitations in hydrogen fermentation, including limited duration time, bacteria leakage, cost, and so on. The use of chloroform treated anaerobic granular sludge as immobilized hydrogen-producing bacteria in an immobilized hydrogen culture may be able to overcome the limitations of traditional immobilization methods. This paper reports the findings on the performance of fed-batch cultures and continuous cultures inoculated with chloroform-treated granules. The chloroform-treated granules were able to be reused over four fed-batch cultures, with pH adjustment. The upflow reactor packed with chloroform-treated granules was studied, and the HRT of the upflow reactor was found to be as low as 4 h without any decrease in hydrogen production yield. Initial pH and glucose concentration of the culture medium significantly influenced the performance of the reactor. The optimum initial pH of the culture medium was neutral, and the optimum glucose concentration of the culture medium was below 20 g chemical oxygen demand/L at HRT 4 h. This study also investigated the possibility of integrating immobilized hydrogen fermentation using chloroform treated granules with immobilized methane production using untreated granular sludge. The results showed that the integrated batch cultures produced 1.01 mol hydrogen and 2 mol methane per mol glucose. Treating the methanogenic granules with chloroform and then using the treated granules as immobilized hydrogen producing sludge demonstrated advantages over other immobilization methods because the treated granules provide hydrogen-producing bacteria with a protective niche, a long duration of an active culture, and excellent settling velocity. This integrated two-stage design for immobilized hydrogen fermentation and methane production offers a promising approach for modifying current anaerobic wastewater treatment processes to harvest hydrogen from the existing systems. PMID- 18418743 TI - Effect of furfural, vanillin and syringaldehyde on Candida guilliermondii growth and xylitol biosynthesis. AB - Xylitol is a five-carbon sugar alcohol with established commercial use as an alternative sweetener and can be produced from hemicellulose hydrolysate. However, there are difficulties with microbiological growth and xylitol biosynthesis on hydrolysate because of the inhibitors formed from hydrolysis of hemicellulose. This research focused on the effect of furfural, vanillin, and syringaldehyde on growth of Candida guilliermondii and xylitol accumulation from xylose in a semi-synthetic medium in microwell plate and bioreactor cultivations. All three compounds reduced specific growth rate, increased lag time, and reduced xylitol production rate. In general, increasing concentration of inhibitor increased the severity of inhibition, except in the case of 0.5 g vanillin per liter, which resulted in a faster late batch phase growth rate and increased biomass yield. At concentrations of 1 g/l or higher, furfural was the least inhibitory to growth, followed by syringaldehyde. Vanillin most severely reduced specific growth rate. All three inhibitors reduced xylitol production rate approximately to the same degree. PMID- 18418744 TI - Production and characterization of biodiesel from tung oil. AB - The feasibility of biodiesel production from tung oil was investigated. The esterification reaction of the free fatty acids of tung oil was performed using Amberlyst-15. Optimal molar ratio of methanol to oil was determined to be 7.5:1, and Amberlyst-15 was 20.8 wt% of oil by response surface methodology. Under these reaction conditions, the acid value of tung oil was reduced to 0.72 mg KOH/g. In the range of the molar equivalents of methanol to oil under 5, the esterification was strongly affected by the amount of methanol but not the catalyst. When the molar ratio of methanol to oil was 4.1:1 and Amberlyst-15 was 29.8 wt% of the oil, the acid value decreased to 0.85 mg KOH/g. After the transesterification reaction of pretreated tung oil, the purity of tung biodiesel was 90.2 wt%. The high viscosity of crude tung oil decreased to 9.8mm(2)/s at 40 degrees C. Because of the presence of eleostearic acid, which is a main component of tung oil, the oxidation stability as determined by the Rancimat method was very low, 0.5h, but the cold filter plugging point, -11 degrees C, was good. The distillation process did not improve the fatty acid methyl ester content and the viscosity. PMID- 18418745 TI - Yeast biomass production in brewery's spent grains hemicellulosic hydrolyzate. AB - Yeast single-cell protein and yeast extract, in particular, are two products which have many feed, food, pharmaceutical, and biotechnological applications. However, many of these applications are limited by their market price. Specifically, the yeast extract requirements for culture media are one of the major technical hurdles to be overcome for the development of low-cost fermentation routes for several top value chemicals in a biorefinery framework. A potential biotechnical solution is the production of yeast biomass from the hemicellulosic fraction stream. The growth of three pentose-assimilating yeast cell factories, Debaryomyces hansenii, Kluyveromyces marxianus, and Pichia stipitis was compared using non-detoxified brewery's spent grains hemicellulosic hydrolyzate supplemented with mineral nutrients. The yeasts exhibited different specific growth rates, biomass productivities, and yields being D. hansenii as the yeast species that presented the best performance, assimilating all sugars and noteworthy consuming most of the hydrolyzate inhibitors. Under optimized conditions, D. hansenii displayed a maximum specific growth rate, biomass yield, and productivity of 0.34 h(-1), 0.61 g g(-1), and 0.56 g l(-1) h(-1), respectively. The nutritional profile of D. hansenii was thoroughly evaluated, and it compares favorably to others reported in literature. It contains considerable amounts of some essential amino acids and a high ratio of unsaturated over saturated fatty acids. PMID- 18418746 TI - Lipase-catalyzed transesterification of rapeseed oil for biodiesel production with tert-butanol. AB - Biodiesel is a fatty acid alkyl ester that can be derived from any vegetable oil or animal fat via the process of transesterification. It is a renewable, biodegradable, and nontoxic fuel. In this paper, we have evaluated the efficacy of a transesterification process for rapeseed oil with methanol in the presence of an enzyme and tert-butanol, which is added to ameliorate the negative effects associated with excess methanol. The application of Novozym 435 was determined to catalyze the transesterification process, and a conversion of 76.1% was achieved under selected conditions (reaction temperature 40 degrees C, methanol/oil molar ratio 3:1, 5% (w/w) Novozym 435 based on the oil weight, water content 1% (w/w), and reaction time of 24h). It has also been determined that rapeseed oil can be converted to fatty acid methyl ester using this system, and the results of this study contribute to the body of basic data relevant to the development of continuous enzymatic processes. PMID- 18418747 TI - Bioethanol production optimization: a thermodynamic analysis. AB - In this work, the phase equilibrium of binary mixtures for bioethanol production by continuous extractive process was studied. The process is composed of four interlinked units: fermentor, centrifuge, cell treatment unit, and flash vessel (ethanol-congener separation unit). A proposal for modeling the vapor-liquid equilibrium in binary mixtures found in the flash vessel has been considered. This approach uses the Predictive Soave-Redlich-Kwong equation of state, with original and modified molecular parameters. The congeners considered were acetic acid, acetaldehyde, furfural, methanol, and 1-pentanol. The results show that the introduction of new molecular parameters r and q in the UNIFAC model gives more accurate predictions for the concentration of the congener in the gas phase for binary and ternary systems. PMID- 18418748 TI - Oxidation in acidic medium of lignins from agricultural residues. AB - Agricultural residues as sugarcane straw and bagasse are burned in boilers for generation of energy in sugar and alcohol industries. However, excess of those by products could be used to obtain products with higher value. Pulping process generates cellulosic pulps and lignin. The lignin could be oxidized and applied in effluent treatments for heavy metal removal. Oxidized lignin presents very strong chelating properties. Lignins from sugarcane straw and bagasse were obtained by ethanol-water pulping. Oxidation of lignins was carried out using acetic acid and Co/Mn/Br catalytical system at 50, 80, and 115 degrees C for 5 h. Kinetics of the reaction was accomplished by measuring the UV-visible region. Activation energy was calculated for lignins from sugarcane straw and bagasse (34.2 and 23.4 kJ mol(-1), respectively). The first value indicates higher cross linked formation. Fourier-transformed infrared spectroscopy data of samples collected during oxidation are very similar. Principal component analysis applied to spectra shows only slight structure modifications in lignins after oxidation reaction. PMID- 18418749 TI - Kinetic modeling and parameter estimation in a tower bioreactor for bioethanol production. AB - In this work, a systematic method to support the building of bioprocess models through the use of different optimization techniques is presented. The method was applied to a tower bioreactor for bioethanol production with immobilized cells of Saccharomyces cerevisiae. Specifically, a step-by-step procedure to the estimation problem is proposed. As the first step, the potential of global searching of real-coded genetic algorithm (RGA) was applied for simultaneous estimation of the parameters. Subsequently, the most significant parameters were identified using the Placket-Burman (PB) design. Finally, the quasi-Newton algorithm (QN) was used for optimization of the most significant parameters, near the global optimum region, as the initial values were already determined by the RGA global-searching algorithm. The results have shown that the performance of the estimation procedure applied in a deterministic detailed model to describe the experimental data is improved using the proposed method (RGA-PB-QN) in comparison with a model whose parameters were only optimized by RGA. PMID- 18418750 TI - Analysis of kinetic and operational parameters in a structured model for acrylic acid production through experimental design. AB - In biotechnological processes, a great number of factors can influence the income productivity and conversion. Normally, it is not evident which of these factors are the most important and how they interact. In this work, multivariate analysis techniques are used as experimental design coupled to a detailed deterministic model to identify the parameters with the most significant impact on the model to represent well the acrylic acid production process. It is proposed as an alternative process, having sugarcane as feedstock, to the petrochemical-based ones that have significant environmental impacts for their production. To increase the competitiveness of renewable acrylic-acid-based process, it is necessary to find out working conditions near the optimal region, which is not an easy task, as the process is multivariable and non-linear. The mapping of the dynamics of the developed process is made using techniques of factorial design together with the methodology of Plackett-Burman. It is shown that it is possible to increase the process performance by choosing optimized conditions for the reactor operation. PMID- 18418751 TI - Optimization of oligosaccharide synthesis from cellobiose by dextransucrase. AB - There is a growing market for oligosaccharides as sweeteners, prebiotics, anticariogenic compounds, and immunostimulating agents in both food and pharmaceutical industries. Interest in novel carbohydrate-based products has grown because of their reduced toxicity and low immune response. Cellobiose is potentially valuable as a nondigestible sugar. The reaction of cellobiose, as an acceptor with a sucrose as a donor, catalyzed by a dextransucrase from Leuconostoc mesenteroides B-512FMCM, produced a series of cellobio oligosaccharides. This production system was optimized using a Box-Behnken experimental design for 289 mM of sucrose and 250 mM of cellobiose and 54 U of the enzyme at pH 5.2 and 30 degrees C, to produce maximum yields of oligosaccharide. PMID- 18418752 TI - Fermentation kinetics for xylitol production by a Pichia stipitis D: xylulokinase mutant previously grown in spent sulfite liquor. AB - Spent sulfite pulping liquor (SSL) contains lignin, which is present as lignosulfonate, and hemicelluloses that are present as hydrolyzed carbohydrates. To reduce the biological oxygen demand of SSL associated with dissolved sugars, we studied the capacity of Pichia stipitis FPL-YS30 (xyl3Delta) to convert these sugars into useful products. FPL-YS30 produces a negligible amount of ethanol while converting xylose into xylitol. This work describes the xylose fermentation kinetics of yeast strain P.stipitis FPL-YS30. Yeast was grown in rich medium supplemented with different carbon sources: glucose, xylose, or ammonia-base SSL. The SSL and glucose-acclimatized cells showed similar maximum specific growth rates (0.146 h(-1)). The highest xylose consumption at the beginning of the fermentation process occurred using cells precultivated in xylose, which showed relatively high specific activity of glucose-6-phosphate dehydrogenase (EC 1.1.1.49). However, the maximum specific rates of xylose consumption (0.19 g(xylose)/g(cel) h) and xylitol production (0.059 g(xylitol)/g(cel) h) were obtained with cells acclimatized in glucose, in which the ratio between xylose reductase (EC 1.1.1.21) and xylitol dehydrogenase (EC 1.1.1.9) was kept at higher level (0.82). In this case, xylitol production (31.6 g/l) was 19 and 8% higher than in SSL and xylose-acclimatized cells, respectively. Maximum glycerol (6.26 g/l) and arabitol (0.206 g/l) production were obtained using SSL and xylose acclimatized cells, respectively. The medium composition used for the yeast precultivation directly reflected their xylose fermentation performance. The SSL could be used as a carbon source for cell production. However, the inoculum condition to obtain a high cell concentration in SSL needs to be optimized. PMID- 18418753 TI - Selective enrichment of a methanol-utilizing consortium using pulp and paper mill waste streams. AB - Efficient utilization of carbon inputs is critical to the economic viability of the current forest products sector. Input carbon losses occur in various locations within a pulp mill, including losses as volatile organics and wastewater. Opportunities exist to capture this carbon in the form of value-added products such as biodegradable polymers. Waste-activated sludge from a pulp mill wastewater facility was enriched for 80 days for a methanol-utilizing consortium with the goal of using this consortium to produce biopolymers from methanol-rich pulp mill waste streams. Five enrichment conditions were utilized: three high methanol streams from the kraft mill foul condensate system, one methanol-amended stream from the mill wastewater plant, and one methanol-only enrichment. Enrichment reactors were operated aerobically in sequencing batch mode at neutral pH and 25 degrees C with a hydraulic residence time and a solids retention time of 4 days. Non-enriched waste activated sludge did not consume methanol or reduce chemical oxygen demand. With enrichment, however, the chemical oxygen demand reduction over 24-h feed/decant cycles ranged from 79 to 89%, and methanol concentrations dropped below method detection limits. Neither the non-enriched waste-activated sludge nor any of the enrichment cultures accumulated polyhydroxyalkanoates (PHAs) under conditions of nitrogen sufficiency. Similarly, the non-enriched waste activated sludge did not accumulate PHAs under nitrogen limited conditions. By contrast, enriched cultures accumulated PHAs to nearly 14% on a dry weight basis under nitrogen-limited conditions. This indicates that selectively enriched pulp mill waste activated sludge can serve as an inoculum for PHA production from methanol-rich pulp mill effluents. PMID- 18418754 TI - Evaluation of cashew apple juice for the production of fuel ethanol. AB - A commercial strain of Saccharomyces cerevisiae was used for the production of ethanol by fermentation of cashew apple juice. Growth kinetics and ethanol productivity were calculated for batch fermentation with different initial sugar (glucose + fructose) concentrations. Maximal ethanol, cell, and glycerol concentrations were obtained when 103.1 g L(-1) of initial sugar concentration was used. Cell yield (Y (X/S)) was calculated as 0.24 (g microorganism)/(g glucose + fructose) using cashew apple juice medium with 41.3 g L(-1) of initial sugar concentration. Glucose was exhausted first, followed by fructose. Furthermore, the initial concentration of sugars did not influence ethanol selectivity. These results indicate that cashew apple juice is a suitable substrate for yeast growth and ethanol production. PMID- 18418755 TI - Atmospheric pressure liquefaction of dried distillers grains (DDG) and making polyurethane foams from liquefied DDG. AB - In this study, dried distillers grains (DDG) was liquefied in acidic conditions at atmospheric pressure, and polyurethane foams were subsequently prepared from the liquefied DDG. Liquefaction was examined over a range of conditions including liquefaction time of 1-3 h, temperature of 150-170 degrees C, sulfuric acid (as catalyst) concentration of 1.0-3.0 wt%, and liquefaction solvent (ethylene carbonate) to DDG ratio of 3:1-5:1. The bio-polyols in the liquefied DDG were rich in hydroxyl groups, which can react with methylene diphenyl diisocyanate (MDI) to form cross-linked polyurethane networks. The biodegradability of the prepared polyurethane foams was also evaluated. This study strives to broaden the application of DDG as a feedstock for bio-polyurethane preparation. PMID- 18418756 TI - Bacterial cellulose production by Acetobacter xylinum strains from agricultural waste products. AB - Bacterial cellulose is a biopolysaccharide produced from the bacteria, Acetobacter xylinum. Static batch fermentations for bacterial cellulose production were studied in coconut and pineapple juices under 30 degrees C in 5-l fermenters by using three Acetobacter strains: A. xylinum TISTR 998, A. xylinum TISTR 975, and A. xylinum TISTR 893. Experiments were carried out to compare bacterial cellulose yields along with growth kinetic analysis. Results showed that A. xylinum TISTR 998 produced a bacterial cellulose yield of 553.33 g/l, while A. xylinum TISTR 893 produced 453.33 g/l and A. xylinum TISTR 975 produced 243.33 g/l. In pineapple juice, the yields for A. xylinum TISTR 893, 975, and 998 were 576.66, 546.66, and 520 g/l, respectively. The strain TISTR 998 showed the highest productivity when using coconut juice. Morphological properties of cellulose pellicles, in terms of texture and color, were also measured, and the textures were not significantly different among treatments. PMID- 18418757 TI - Overview of special session B--compositional and structural analysis of biomass. AB - Special Session B at the 29th Symposium on Biotechnology for Fuels and Chemicals was the first invited session at this symposium devoted to analytical methods. The special topic was added in response to numerous requests for information on new and innovative methods that could be applied in the growing renewable fuels industry. Presentation topics include analytical methods for the characterization and analysis of maize traits, tools for investigating cell wall limitations to enzymatic degradation, methods for customizing enzyme cocktails for biomass, new techniques for the analysis of carbohydrates, analytical methods that enhance our understanding of pretreatment, improved methods for monitoring process intermediates, and published standard analytical methods for biomass conversion processes. PMID- 18418758 TI - What can be learned from silage breeding programs? AB - Improving the quality of cellulosic ethanol feedstocks through breeding and genetic manipulation could significantly impact the economics of this industry. Attaining this will require comprehensive and rapid characterization of large numbers of samples. There are many similarities between improving corn silage quality for dairy production and improving feedstock quality for cellulosic ethanol. It was our objective to provide insight into what is needed for genetic improvement of cellulosic feedstocks by reviewing the development and operation of a corn silage breeding program. We discuss the evolving definition of silage quality and relate what we have learned about silage quality to what is needed for measuring and improving feedstock quality. In addition, repeatability estimates of corn stover traits are reported for a set of hybrids. Repeatability of theoretical ethanol potential measured by near-infrared spectroscopy is high, suggesting that this trait may be easily improved through breeding. Just as cell wall digestibility has been factored into the latest measurements of silage quality, conversion efficiency should be standardized and included in indices of feedstock quality to maximize overall, economical energy availability. PMID- 18418759 TI - Permethylation linkage analysis techniques for residual carbohydrates. AB - Permethylation analysis is the classic approach to establishing the position of glycosidic linkages between sugar residues. Typically, the carbohydrate is derivatized to form acid-stable methyl ethers, hydrolyzed, peracetylated, and analyzed by gas chromatography-mass spectrometry. The position of glycosidic linkages in the starting carbohydrate are apparent from the mass spectra as determined by the location of acetyl residues. The completeness of permethylation is dependent upon the choice of base catalyst and is readily confirmed by matrix assisted laser desorption/ionization time-of-flight mass spectrometry mass spectrometry. For the permethylation of beta-cyclodextrin, Hakomori dimsyl base is shown to be superior to the NaOH-dimethyl sulfoxide system, and the use of the latter resulted in selective under-methylation of the 3-hydroxy groups. These techniques are highly applicable to residual carbohydrates from biofuel processes. PMID- 18418767 TI - Improving natural killer cells. PMID- 18418768 TI - Data management in the cell therapy production facility: the batch process record (BPR). AB - The activities of cell therapy establishments are associated with substantial amounts of information. For reasons of best practice, regulation and adherence to prevailing standards, the data generated in the course of cell therapy product processing must be recorded and retained in an organized manner. Because cell therapy products are functionally pharmaceuticals, the paradigm of the pharmaceutical manufacturing batch process record (BPR) is proposed as a unit for collecting the data resulting from processing. Considerations for cell-processing facilities for the design of BPR and possible selection of electronic data recording tools are reviewed, including data to collect in response to regulatory or accreditation mandates and different types of electronic data management tools that may be employed. Additionally, considerations for selection, qualification and validation of computer software for maintenance of the BPR are addressed. PMID- 18418769 TI - No increased trapping of multipotent mesenchymal stromal cells in bone marrow filters compared with other bone marrow cells. AB - BACKGROUND: Multipotent mesenchymal stromal cells (MSC) are candidates for cellular therapy in regenerative medicine and as treatment of graft-versus-host disease (GvHD) after hematopoietic stem cell (HSC) transplantation. It has been suggested that MSC may be trapped in bone marrow (BM) filters during the stem cell procurement and lost from the HSC graft. METHODS: We investigated filtered BM and filters from six HSC donors. MSC were expanded from the two sources and investigated by flow cytometry, doubling capacity, differentiation ability and suppression in mixed lymphocyte cultures. RESULTS: A range of 0.3-3.4% cells was trapped in the filters. By flow cytometry, there was no difference in the proportions of different cell types between the filter-retrieved and filtered BM cells. The phenotype, immunosuppressive capacity, differentiation and growth were equal in MSC expanded from the two cell sources. DISCUSSION: Given the low number of trapped cells, filters do not appear to be a good source of MSC. When intended for clinical transplantation, MSC need to be expanded ex vivo to achieve sufficient doses for a clinical effect. PMID- 18418770 TI - Immunophenotype characterization of rat mesenchymal stromal cells. AB - BACKGROUND: Mesenchymal stromal cells (MSC) have shown diverse therapeutic potential. While characterization of human and mouse MSC has seen significant advances, rat bone marrow-derived MSC (rBM-MSC) remain under-characterized. We detail the isolation, expansion, differentiation, and detailed immunocharacterization of rBM-MSC. METHODS: Rat MSC were isolated and expanded in multipotent adult progenitor cell (MAPC) media, and cell-surface marker expression through 10 passages was used to characterize the population and multipotency was confirmed via differentiation. RESULTS: By passage 3, rBM-MSC were found to be CD11b-, CD45-, CD29+, CD49e+, CD73+, CD90+, CD105+ and Stro-1+, without the use of cell sorting. Media selection was responsible for the isolation of a nearly homogeneous population of rBM-MSC. The rBM-MSC immunophenotype changed by passage 10, showing decreases in CD73, CD105 and Stro 1 expression. DISCUSSION: Detailed characterization of cell populations facilitates accurate and reproducible cell therapy investigation. Given the expanding body of research involving rBM-MSC, these results advance our ability to compare rBM-MSC populations. PMID- 18418771 TI - Dynamic distribution of bone marrow-derived mesenchymal stromal cells and change of pathology after infusing into mdx mice. AB - BACKGROUND: Mesenchymal stromal cells (MSC) are attractive candidates for the treatment of Duchenne muscular dystrophy (DMD) but how the donor MSC distribute in multiple organs and whether the increased dystrophin leads to a change in the pathology of mdx mice is still uncertain. In this research we detected the distribution of MSC and the pathology of mdx mice after MSC infusion. METHODS: MSC were isolated from rat bone marrow (BM) and expanded in proliferation medium. MSC of the fifth passage were delivered intravenously into irradiated mdx mice. The distribution of MSC labeled by [3H]TdR into a recipient's organs was calculated by radioactivity. The expression of dystrophin was detected at weeks 4, 8, 12 and 16 after MSC transplantation by immunofluorescence staining, RT-PCR and Western blot. Serum creatine kinase (CK) and centrally nucleated fiber (CNF) were also detected to assess the change in pathology. RESULTS: 24-48 h after transplantation, MSC were mainly found in the BM, liver and lung. The radioactivity in these organs decreased, whereas skeletal and myocardial muscle radioactivity increased gradually over time. In accordance with the increased radioactivity in skeletal muscle, the amount of dystrophin-positive myofibers increased. Furthermore, serum CK and CNF decreased slightly, suggesting specific pathophysiologic features of the dystrophic muscle were partially restored. DISCUSSION: Upon certification of the distribution of transplanted MSC in irradiated mdx mice, we found evidence of myogenic differentiation of MSC in skeletal muscle. This research may help us understand the mechanism of therapy of MSC transplantation. PMID- 18418772 TI - Functional characterization of interleukin-15 gene transduction into the human natural killer cell line NKL. AB - BACKGROUND: Genetic modification of natural killer (NK) cells is a potential approach to gene-based immunotherapy of cancer. We created human interleukin-15 (hIL-15) gene-modified NKL cells and investigated their functional characterization in vitro. METHODS: A recombinant vector (pcDNA3-IL15) or control vector (pcDNA3) was transferred into NKL cells by an electroporation method. Standard reverse transcriptase-polymerase chain reaction (RT-PCR), flow cytometry and MTT methods were performed for NK cell proliferation, apoptosis, cytotoxicity assays and gene expression tests. RESULTS: Compared with parental NKL cells, hIL 15 gene modification promoted NK cell proliferation at low doses of IL-2 and inhibited cell apoptosis, which was associated with the up-regulation of anti apoptosis genes Bcl-2, Bcl-xl and Mcl-1 as well as the down-regulation of apoptosis genes Bim and Noxa. Moreover, the anti-tumor activity of hIL-15 gene transduced NKL cells against human hepatoma cancer cell line HepG2, H7402 and PLC/PRF-5 cells was enhanced, at least partly, through increasing expression of cytotoxicity-associated genes, including interferon (IFN)-gamma, perforin and FasL. DISCUSSION: The hIL-15 genetic modification could improve the proliferation, anti-apoptosis and natural cytotoxicity of NKL cells against hepatocarcinoma cells. These data suggest that hIL-15 gene-modified NKL cells could be useful for clinical cancer immunotherapy in the future. PMID- 18418773 TI - Alloreactivity of ex vivo-expanded T cells is correlated with expansion and CD4/CD8 ratio. AB - Background We have demonstrated previously that retroviral-mediated transfer of a suicide gene into bone marrow (BM) donor T cells allows an efficient control of graft-versus-host disease (GvHD) after allogeneic BM transplantation. However, the 12 days of ex vivo culture required for the production of gene-modified cells (GMC), including soluble CD3 monoclonal antibody (MAb)-mediated activation and expansion with interleukin (IL)-2, induced a decrease of GMC alloreactivity and a reversal of their CD4/CD8 ratio. Improving the culture protocol in order to maintain the highest alloreactivity is of critical importance in obtaining an optimal graft-versus-leukemia (GvL) effect. Methods Peripheral blood mononuclear cells were activated with soluble CD3 MAb or CD3 and CD28 MAb co-immobilized on beads and expanded for 12 days in the presence of IL-2, IL-7 or IL-15 before analysis of alloreactivity and phenotype. Results Replacing the CD3 MAb by CD3/CD28 beads led to similar in vitro alloreactivity but improved the expansion and in vivo alloreactivity of GMC. Replacing the IL-2 with IL-7, but not IL-15, or decreasing IL-2 or IL-7 concentrations, improved the in vitro alloreactivity of expanded cells but was associated with lower expansion. Indeed, the alloreactivity of expanded cells was negatively correlated with cell expansion and positively correlated with CD4/CD8 ratio and CD8 expression level. Discussion Quantitative (i.e. low CD4/CD8 ratio) and qualitative (e.g. low CD8 expression) defects may account for the decreased alloreactivity of GMC. Using CD3/CD28 beads and/or IL-7 is more beneficial than CD3 MAb and IL-2 for preventing perturbations of the alloreactivity and phenotype of GMC. PMID- 18418774 TI - Detailed analysis of cytomegalovirus (CMV)-specific T cells expanded for adoptive immunotherapy of CMV infection following allogeneic stem cell transplantation for malignant disease. AB - BACKGROUND: Cytomegalovirus (CMV) infection and its treatment causes significant morbidity following allogeneic stem cell transplantation (SCT) for malignancies. We studied the phenotype, function and growth kinetics of CMV pp65 antigen (Ag) specific T cells expanded in a short-term culture for adoptive therapy. METHODS: Peripheral blood mononuclear cells (PBMC) from CMV-seropositive donors were cultured in various conditions with CMV pp65((495-503)) peptide to determine the most effective method for generating CMV-specific T cells. CMV-expanded cultures were tested for frequency, phenotype and functionality using peptide-MHC tetramer analysis, cytokine flow cytometry and cytolytic assays. A patient undergoing allogeneic SCT was administered CMV pp65-specific T cells generated from the donor based on these data, and recipient PBMC were analyzed following T-cell infusion. RESULTS: CMV pp65-specific T cells were consistently generated from CMV seropositive donors at high frequencies (20-40% of CD8+ T cells), secreted interferon-gamma (IFN-gamma) in response to CMV peptide and had lytic activity against CMV peptide-expressing targets. Cultured CMV-specific T cells were infused into a SCT recipient without toxicity. DISCUSSION: Stimulating donor PBMC to generate functional, Ag-specific T cells for infusion into SCT recipients was accomplished consistently using readily available technology. We observed no toxicity in one patient receiving T cells and were able to monitor infused cells. These findings support further study of this approach as a prophylaxis against the risk of infection in patients receiving allogeneic transplantation from CMV seropositive donors. PMID- 18418775 TI - Mobilization of CD34+ hematopoietic cells, colony-forming cells and long-term culture-initiating cells into the peripheral blood of patients with an acute cerebral ischemic insult. AB - BACKGROUND: In vitro and in vivo data indicate that stem cells found in the bone marrow (BM) are capable of differentiating into neural cells. The aim of this study was to investigate whether potentially pluripotent hematopoietic stem and progenitor cells are recruited from the BM into the peripheral blood as a reaction to ischemic damage of neural tissues. MATERIALS: The number of CD34+ cells, colony-forming cells (CFC) and long-term culture-initiating cells (LTC-IC) was measured within 24 h and on day 7 after stroke onset by flow cytometry, or in functional assays in the peripheral blood of 10 patients with acute middle cerebral artery infarct. The National Institute of Health stroke scale, Barthel index and modified Rankin scale were used to monitor the clinical outcome. RESULTS: In four patients receiving intravenous thrombolytic therapy (tissue plasminogen activator; TPA), no significant increase of CD34+ cells, CFC or LTC IC was detected. In six patients without thrombolytic treatment, the mean number of CD34+ cells/mL increased significantly from 1181+/-248 at day 1 to 3001+/-881 at day 7. Accordingly, the numbers of CFC and LTC-IC increased 2.7- and 1.7-fold. Granulocyte colony-stimulating factor and neutrophil elastase were monitored by ELISA and remained unchanged during the study period. DISCUSSION: Our results showed a recruitment of hematopoietic progenitor cells from the BM into the peripheral blood after acute ischemic stroke when no thrombolytic treatment was given. Increased progenitor cell recruitment might be caused by so far unknown signaling stimuli of the ischemic penumbra for stem cell mobilization. PMID- 18418776 TI - Investigational new drugs submitted to the Food and Drug Administration that are placed on clinical hold: the experience of the Office of Cellular, Tissue and Gene Therapy. AB - BACKGROUND: Cell and gene therapies are medical products regulated by the U.S. Food and Drug Administration (FDA) within its Center of Biologics Evaluation and Research (CBER) in the Office of Cellular, Tissue, and Gene Therapy (OCTGT). Clinical research using cell and gene therapies in the United States must be conducted under an Investigational New Drug (IND) application. After an initial, 30-day review FDA either places an IND on clinical hold or allows the IND to proceed. METHODS: We reviewed letters sent by OCTGT to IND sponsors that were placed on clinical hold. We categorized each deficiency and determined its frequency. RESULTS: We found that similar deficiencies existed across IND applications and we tabulated the most common deficiencies. DISCUSSION: We discussed the deficiencies and the resources that can help individuals avoid those deficiencies. We believe that awareness of the common deficiencies along with the applicable resources can reduce the frequency of clinical holds and allow clinical studies to proceed without delay. We also believe that this information will guide the FDA as to how to facilitate development of safe and effective cell and gene therapies. PMID- 18418778 TI - What's new in nicotine & tobacco research? PMID- 18418779 TI - Correlates of cigarette smoking during pregnancy and its genetic and environmental overlap with nicotine dependence. AB - Cigarette smoking during pregnancy (CSDP) is associated with a number of negative outcomes in the offspring. Therefore, clarifying the correlates of CSDP and the extent to which CSDP is associated with nicotine dependence is an important step toward reducing its rate in the general population. Using data from 1,134 adult Australian female monozygotic and dizygotic twin pairs, we explored the associations between CSDP and sociodemographic and psychiatric correlates and between CSDP and patterns of cigarette smoking. Further, we examined the role of heritable and environmental influences on CSDP and investigated whether these latent risk factors are shared with a predisposition to nicotine dependence. Women smoking during an entire pregnancy reported heavier dependence and more unsuccessful quit attempts, compared with the community sample of mothers and with women who smoked during only part of a pregnancy. Educational attainment, weekly church attendance, spousal current smoking, and nicotine dependence also were associated with CSDP. Heritable influences explained 34% of the variation in CSDP, with the remainder related to nonshared environmental factors. A large proportion of the genetic influences on CSDP were shared with DSM-III-R nicotine dependence, with little overlap across the nonshared environmental influences. A lifetime history of difficulty with smoking cessation, in conjunction with social background and psychiatric comorbidity, especially during pregnancy, needs to be considered by treatment providers when counseling expectant mothers about the potential risks of CSDP. PMID- 18418780 TI - Influence of American acculturation on cigarette smoking behaviors among Asian American subpopulations in California. AB - Using combined data from the population-based 2001 and 2003 California Health Interview Surveys, we examined ethnic and gender-specific smoking behaviors and the effect of three acculturation indicators on cigarette smoking behavior and quitting status among 8,192 Chinese, Filipino, South Asian, Japanese, Korean, and Vietnamese American men and women. After adjustment for potential confounders, current smoking prevalence was higher and the quit rate was lower for Korean, Filipino, and Vietnamese American men compared with Chinese American men. Women's current smoking prevalence was lower than men's in all six Asian American subgroups. South Asian and Korean American women reported lower quit rates than women from other ethnic subgroups. Asian American men who reported using only English at home had lower current smoking prevalence and higher quit rates, except for Filipino and South Asian American men. Asian American women who reported using only English at home had higher current smoking prevalence except for Japanese women. Being a second or later generation immigrant was associated with lower smoking prevalence among all Asian American subgroups of men. Less educated men and women had higher smoking prevalence and lower quit rates. In conclusion, both current smoking prevalence and quit rates vary distinctively across gender and ethnic subgroups among Asian Americans in California. Acculturation appears to increase the risk of cigarette smoking for Asian American women. Future tobacco-control programs should target at high-risk Asian American subgroups, defined by ethnicity, acculturation status, and gender. PMID- 18418781 TI - The effect of subjective normative social images of smokers on children's intentions to smoke. AB - This paper expands the literature on peers' influence on a youth's behaviors through the examination of the effect of subjective normative social images of smokers (perceived peers' social images of smokers) on subsequent intentions to smoke and the relation between subjective normative social images and the youth's own social images, or prototypes. Data are from the two oldest cohorts (4th and 5th graders at the first assessment) and from the first five assessments of the Oregon Youth Substance Use Project, an ongoing longitudinal study. Results showed that both children's subjective normative social images and prototypes uniquely predicted intentions to smoke cigarettes at the subsequent assessment. Across time, subjective normative social images were more positive than the youth's own prototypes, and subjective normative social images and children's own prototypes were reciprocally related over time. Results provide support for subjective normative social images as a risk factor for children's smoking and suggest targeting this mechanism in smoking prevention programs for children. PMID- 18418782 TI - Severity of nicotine dependence moderates performance on perceptual-motor tests of attention. AB - Acute abstinence from cigarette smoking by nicotine-dependent smokers has been linked with cognitive deficits, but the role of nicotine dependence per se in these effects is not known. We therefore tested the relationships of nicotine dependence and smoking history with performance in perceptual-motor, timed tests of attention. Nicotine-dependent smokers (n = 37) and nonsmokers (n = 48), 18-55 years old, took both the d2 Test of Attention and the Digit Symbol Test on each of 2 test days. For smokers, testing on one day began after ad libitum smoking (<45 min since last cigarette); and on the other day, it began after overnight abstinence (>13 hr since last cigarette). On each test day, there were two test blocks with an intervening break, when only the smokers each smoked one cigarette. There were no significant effects of abstinence or of smoking one cigarette on the performance of smokers; however, across conditions, the smokers' performance on both tests correlated negatively with severity of nicotine dependence but not lifetime cigarette consumption or cigarette craving. Smokers with high nicotine dependence performed more slowly on both tests than less dependent smokers or nonsmokers. The findings suggest that severity of nicotine dependence and slowness in perceptual-motor tasks of attention share an underlying basis. PMID- 18418783 TI - Diagnostic accuracy of NicAlert cotinine test strips in saliva for verifying smoking status. AB - Semiquantitative immunoassay technology, in the form of rapid test strips, offers a less time-consuming and less costly alternative to other methods of verifying self-reported smoking status, such as gas chromatography-nitrogen phosphorus detection (GC). Unfortunately, information on the validity and reliability of some test strips in urine and saliva samples is not always available. This paper describes the diagnostic accuracy of one type of test strip currently available (NicAlert cotinine test strips; NCTS). GC was used as the reference standard and saliva as the sample medium. The study involved 86 people (41 smokers and 45 nonsmokers) aged 18 years or over, who were able to understand written English and provide written consent. Pregnant women, women with infants less than 6 weeks old, and people who had eaten 30 min prior to sample collection were excluded. Two saliva samples were collected simultaneously from each participant, with one sample tested using NCTS and the other by GC analysis. People with at least 10 ng/ml cotinine (in both tests) in their saliva were considered smokers. NCTS were found to have a specificity of 95% (95% CI 89%-100%), a sensitivity of 93% (95% CI 85%-100%), a positive predictive value of 95% (95% CI 89%-100%), and a negative predictive value of 93% (95% CI 86%-100%). The use of NCTS is a valid and reliable method, compared with GC, to test saliva samples for verification of smoking status. PMID- 18418784 TI - Tobacco industry research and efforts to manipulate smoke particle size: implications for product regulation. AB - Over the past half-century of cigarette design, tobacco manufacturers have prioritized efficiency of delivery alongside ease of inhalation and use. As a result, the modern cigarette is uniquely effective at facilitating the absorption of nicotine as well as carcinogens and other toxins. The present study draws on internal tobacco company documents to assess industry consideration of the role of smoke particle size as a potentially controllable influence over inhalation patterns and lung exposure. Tobacco manufacturers evaluated particle size manipulation both as a means of controlling physical and sensory product attributes and as a possible approach to reducing health hazards related to exposure. Industry scientists concluded that the smoke aerosol particle distribution of conventional cigarettes, constructed within common parameters, falls within a narrow and effective inhalation range. However, the internal findings suggest that differences in smoke particle size distribution are possible through less conventional approaches to product design. We propose that particle size be included among the many design features to be considered in emerging tobacco product regulation. However, the present review does not address whether particle size regulation would be a plausible means of substantially reducing addictiveness or harmfulness of cigarettes, and therefore we do not propose it as a high-priority target for regulation. PMID- 18418785 TI - Anxiety sensitivity and perceived control over anxiety-related events: associations with smoking outcome expectancies and perceived cessation barriers among daily smokers. AB - The present investigation examined the extent to which the cognitive factors of anxiety sensitivity (AS) and perceived control over anxiety-related events are independently related to smoking outcome expectancies and perceived barriers to quitting. Participants were 125 community-recruited adult, daily smokers. Consistent with hypotheses, AS and perceived control over anxiety-related events independently and significantly predicted smokers' expectancies for negative affect reduction from smoking, whereas only AS predicted expectancies for negative personal consequences from smoking. Also as hypothesized, AS and perceived control over anxiety-related events each independently and significantly predicted level of general perceived barriers to quitting smoking. All of the observed significant effects were evident above and beyond the variance accounted for by gender, alcohol consumption, number of cigarettes smoked per day, and negative affectivity. Together, these findings further the literature on the relation between anxiety-relevant cognitive factors and psychological smoking processes. PMID- 18418786 TI - Impact of smoking cessation and lifetime exposure on C-reactive protein. AB - C-reactive protein (CRP) levels predict coronary heart disease (CHD) risk. Levels are raised among smokers, but the effect of smoking cessation is unclear. Exposure to secondhand smoke (SHS) may be a confounder. Lifetime smoking exposure may have a dose effect on CRP among smokers, but it is unclear if this persists after cessation. We analyzed cross-sectional data on 4,072 adults recruited to a Scotland-wide population health survey who did not have CHD and were not on nicotine replacement therapy. CRP fell with time from cessation but was still raised up to 5 years after adjustment for case-mix (p<.001). SHS exposure was greater among ex-smokers than never-smokers (median cotinine 0.5 ng/ml vs. 0.4 ng/ml, p<.001) but did not explain the difference. Among smokers, there was a dose relationship between pack years and CRP on both univariate, F(4,1279) = 31.841, p<.001, and multivariate, F(4,1085) = 3.499, p = .008, analysis. Among ex smokers there was also a dose relationship between pack-years and CRP, F(4,751) = 14.108, p<.001, which was independent of time from cessation and case-mix, F(4,466) = 3.744, p = .005. That CRP does not fall to normal levels immediately and that lifetime smoking exposure continues to impact on CRP levels post cessation suggest that CRP is not raised as a direct effect of cigarette smoke but rather via a secondary mechanism, such as tissue damage causing an inflammatory stimulus. Our results reinforce the need to encourage smoking cessation as early as possible. PMID- 18418787 TI - Reliability and validity of a Short Form of the Tobacco Craving Questionnaire. AB - The Tobacco Craving Questionnaire (TCQ) is a valid and reliable 47-item self report instrument that assesses tobacco craving in four dimensions: emotionality, expectancy, compulsivity, and purposefulness. For use in research and clinical settings, we constructed a 12-item version of the TCQ by selecting three items from each of the four factors that exhibited optimal within-factor reliability (Cronbach's alpha coefficient) and inter-item correlation. Smokers (N = 196) completed the TCQ-Short Form (TCQ-SF) after overnight tobacco deprivation and on a separate day during ad libitum smoking. Confirmatory factor analyses indicated acceptable model fit for a 4-factor model, with congruence coefficients suggesting high to very high similarity in factor patterns and magnitude of factor loadings between the TCQ and TCQ-SF in both conditions. Scores on each factor were significantly greater after tobacco deprivation than ad libitum smoking, were associated with measures of tobacco withdrawal, and varied with degree of nicotine dependence. Cronbach's alpha coefficients and average inter item correlations were similar in both conditions and were consistent with reliability values obtained in the initial validation of the TCQ. Test-retest correlation coefficients were also similar to those found in a previous study. These findings suggest that the TCQ-SF is as valid and reliable as the 47-item TCQ in measuring tobacco craving. PMID- 18418788 TI - A formative examination of messages that discourage tobacco use among junior enlisted members of the United States military. AB - Smoking prevalence among junior enlisted members of the United States armed forces significantly exceeds that of civilians. Furthermore, nearly 40% of junior enlisted who smoke reported they initiated smoking after joining the military. Although the tobacco industry has attempted to develop messages that encourage military members to use tobacco, no research has examined messages that would counteract industry messages and effectively discourage military members from using tobacco. This study conducted 24 focus groups on four Air Force and two Army installations (N = 189 personnel) to discover effective messages that discourage tobacco use among junior enlisted personnel. Four message themes were identified that hold promise for tobacco control efforts in the military: (a) smoking harms one's ability to positively influence others, (b) smoking increases the chance a military member will be discharged from the military prematurely, (c) smoking lowers readiness to fight and win wars, and (d) smokers are not as productive as other military personnel. In contrast, messages focusing on manipulation by the tobacco industry and the health effects of tobacco use were not strongly supported by military personnel. PMID- 18418789 TI - Environmental tobacco smoke exposure among Korean American nonsmokers in California. AB - Information about the extent and patterns of environmental tobacco smoke (ETS) exposure among Korean Americans is sparse, despite the population's having one of the highest male smoking rates. This paper estimates the prevalence of ETS exposure among Korean American nonsmokers in California, and identifies demographic and other characteristics associated with exposure. Data were collected during 2001-2002 from telephone interviews (in English or Korean) with 2,328 nonsmoking Korean American adults. ETS was encountered by 31% of respondents during a typical day. Exposure was most common in "other locations," where 24% of respondents were exposed, compared with 6% at home and 9% at work. Among those exposed, the greatest dose of exposure occurred at work (6 cigarettes/day) and at home (5 cigarettes/day). Women were four times more likely than men to be exposed to ETS at home (8% vs. 2%, respectively). For both men and women, the odds of exposure were greater among those who were younger, who were unmarried, and whose friends smoked. Additionally, traditional men and bicultural women had greater odds of ETS exposure than those who were more acculturated. Women who were married to smokers, had no children at home, consumed more alcohol, and had no home smoking ban also had greater likelihood of exposure. The results indicate the need for a complete ban of smoking in workplaces and in private homes to prevent exposure, particularly for women whose husbands smoke. PMID- 18418790 TI - Smoking is associated with lower performance in WAIS-R Block Design scores in adults with ADHD. AB - Adults with attention-deficit/hyperactivity disorder (ADHD) are predisposed to smoking, but the neuropsychological correlates of this association have not been elucidated so far. The present study evaluates possible associations between cognitive performance and smoking and other comorbidities in adults with ADHD. Two hundred and sixty-four (264) patients were evaluated in the adult ADHD outpatient clinic of the Hospital de Clinicas de Porto Alegre. The diagnoses were based on the DSM-IV criteria and interviews were performed with the Portuguese version of K-SADS-E for ADHD and oppositional-defiant disorder. Axis I psychiatric comorbidities were evaluated with the SCID-IV and the cognitive performance with the Vocabulary and Block Design subtests of the Wechsler Adult Intelligence Scale-Revised (WAIS-R). The evaluation of the influence of the WAIS R scores on each dependent variable was performed with logistic regression analyses. Lower scores in the Block Design subtest of WAIS-R were associated with smoking and the presence of anxiety disorder. These results suggest that a subgroup of ADHD patients with lower Block Design subtest scores may be at increased risk of smoking as a cognitive enhancement. Our findings also confirmed the previously suggested association between anxiety and lower Block Design scores. PMID- 18418791 TI - Exercise as an adjunct to nicotine gum in treating tobacco dependence among women. AB - This was the first randomized, controlled smoking cessation trial assessing the efficacy of an exercise intervention as an adjunct to nicotine gum therapy in comparison with both equal contact control and standard care control conditions. Sedentary female smokers aged 18-55 years were provided with nicotine gum treatment along with brief behavioral counseling and were randomized into one of these three behavioral adjunct conditions. In the "intent-to-treat" sample (N = 182), at end of treatment and at 1-year follow-up, there were clear, but nonsignificant, trends in univariate analyses in which the exercise and equal contact control conditions both had higher rates of abstinence than the standard care control. However, when adjusting for other predictors of relapse in a multiple logistic regression, both exercise and equal contact control showed an advantage over standard care control in avoiding early relapse (i.e., after 1 week). In a multivariate survival model adjusting for other predictors, the equal contact condition had a significantly lower likelihood of relapse compared with the standard care condition and there was a near significant trend in which exercise offered an advantage over standard care as well. While these findings suggest a slightly improved likelihood of abstinence with exercise compared with standard care, exercise did not differ from equal contact control in its efficacy. Potential explanations for these equivalent levels of efficacy and implications for the findings are discussed. PMID- 18418792 TI - The menthol smoker: tobacco industry research on consumer sensory perception of menthol cigarettes and its role in smoking behavior. AB - The use of menthol in cigarettes is actively promoted by the tobacco industry for its perceived sensory benefits, and smokers of menthol cigarettes commonly differ from nonmenthol smokers in markers of smoking behavior and addiction. In this study, we analyzed internal tobacco industry documents to describe the relationships between sensory perception and the attitudes, preferences, and patterns of cigarette use among menthol smokers. Two unique types of menthol smoker emerged from this analysis: those who cannot tolerate the harshness and irritation associated with smoking nonmenthol cigarettes, and those who seek out the specific menthol flavor and associated physical sensation. Among the first segment of menthol smokers, menthol reduces negative sensory characteristics associated with smoking. This segment of smokers may include a large proportion of occasional smokers or young people, as well as smokers who have "traded down" to a less strong cigarette because of perceived harshness or negative health effects. Some established menthol smokers, on the other hand, appear to be tolerant of and even actively seek stronger sensory attributes, including higher menthol levels. Smokers of these "stronger" menthols have traditionally been disproportionately Black and male. Some beginning or occasional smokers may adopt menthols for their mild properties and to cover up the taste of tobacco, but then develop a stronger desire for the menthol taste over time. Future research measuring smoking behavior and evaluating cessation outcomes of menthol smokers should consider the duration of menthol use and differentiate smokers according to their reasons for using menthols. PMID- 18418793 TI - A randomized controlled clinical trial of bupropion SR and individual smoking cessation counseling. AB - Efficacy of bupropion SR and individual counseling as smoking cessation treatments was assessed in a randomized, placebo-controlled clinical trial among adult daily smokers. Bupropion SR treatment and counseling were fully crossed in this factorial design so that the efficacy of each treatment and the combination could be estimated, relative to a placebo medication and assessment control condition. Intent-to-treat analyses indicated that bupropion SR increased abstinence rates at the end of treatment, relative to the placebo medication conditions, for both biochemically confirmed 7-day point-prevalence abstinence (OR = 1.97, 95% CI 1.04-3.72) and self-reported prolonged abstinence (OR = 2.90, 95% CI 1.66-5.06). Bupropion SR treatment also improved latency to lapse and relapse and improved the latency between lapse and relapse in survival analyses. Medication effects were more modest for both 12-month point-prevalence abstinence (OR = 1.47, 95% CI 0.74-2.92) and prolonged abstinence (OR = 1.34, 95% CI 0.66 2.72). Counseling was not associated with increases in the likelihood of abstinence at any time point (odds ratios ranged from 0.80 to 1.16 across abstinence outcomes in the full intent-to-treat sample). Counseling and medication did not significantly interact at any time point, and adding counseling did not improve end-of-treatment point-prevalence abstinence (OR = 1.17, 95% CI 0.68-2.03) or prolonged abstinence (OR = 1.26, 95% CI 0.75-2.12) substantially when offered in conjunction with active medication. PMID- 18418794 TI - Cigarette smoking is an independent predictor of nonadherence in HIV-infected individuals receiving highly active antiretroviral therapy. AB - Current treatment guidelines for the management of HIV-infected individuals emphasize the importance of excellent adherence to antiretroviral medications. We conducted a prospective 24-week study of adherence to lopinavir/ritonavir in a group of 64 subjects using the Medication Event Monitoring System (MEMS). A range of demographic and clinical information, including cigarette smoking status, was collected from each participant. The overall mean adherence rate of the study cohort was 72.8% (SD = 22.2%). Current smokers took 63.5% (SD = 22.1) of prescribed doses, compared with 84.8% (SD = 15.8%) in nonsmokers (p<.001). We found no difference in adherence rates between ex-smokers and subjects who had never smoked. In a multiple linear regression model, factors independently associated with lower adherence rates included current smoking (p = .001), lower CD4+ lymphocyte count at enrollment (p = .04), and lower educational attainment (p = .04). Depression and history of illicit substance use were not associated with nonadherence. In our study cohort, current cigarette smoking was an important and significant marker of inferior adherence to antiretroviral medication. PMID- 18418795 TI - Smoking behavior, knowledge, attitudes and practice among health care providers in Changsha city, China. AB - With the smoking epidemic in China, the role that health care providers (HCPs) could play in tobacco control will be critical. As a preliminary step, this study identified smoking behavior, knowledge, attitudes, and practice (KAP) regarding smoking and smoking control among HCPs in Changsha city, China. The study design was cross-sectional. The instruments - Demographic Information Scale and KAP Scale - were distributed to 420 physicians and 400 nurses in four hospitals in Changsha city. The effective response rate was 77.56% (636/820). Among the respondents, 20.8% were current smokers, which represented 35.7% of the physicians and 1.4% of the nurses, and 43.0% of the males and 1.2% of the females. The mean knowledge score was 22.3+/-4.4 (n = 30). Knowledge regarding smoking's harmful effects and smoking-related respiratory system diseases was high. Nonetheless, tobacco addiction was underestimated. The mean attitudes and practice score was 17.7+/-2.3, 15.1+/-3.4 (n = 20). Eighty-seven percent approved their role in smoking control, but only 45% HCPs practically informed patients about methods of smoking cessation. Smoking behavior was negatively associated with KAP, and 59.7% of the HCPs thought that inadequate knowledge affected their practice. In conclusion, HCPs had some knowledge and positive attitudes regarding the cessation of smoking, but were relatively unoptimistic regarding putting smoking control into practice. The high smoking rate among male HCPs and knowledge insufficiency about smoking may pose barriers in tobacco control efforts for HCPs. As such, interventions for the cessation of smoking, and continual education on smoking control for HCPs, are indicated in China. PMID- 18418797 TI - Emergence of a new clone carrying Panton-Valentine leukocidin genes and staphylococcal cassette chromosome mec type V among methicillin-resistant Staphylococcus aureus in Greece. AB - Clonal analysis and PCR screening for the presence of Panton-Valentine leukocidin (PVL) genes was performed among 694 methicillin-resistant Staphylococcus aureus (MRSA) cases collected during a 2-y period in Greece. The detection rate of PVL positive MRSA is high, both in the community and in hospital. Clonal analysis revealed the predominance among the PVL-positive strains of the clonal complex CC80 (ST80-IV) and the emergence of ST377 clone carrying agr1 allele and SCCmec type V. PMID- 18418798 TI - Comparison of the interferon- gamma release assay and the tuberculin skin test for contact investigation of tuberculosis in BCG-vaccinated health care workers. AB - Health care workers are at increased risk of Mycobacterium tuberculosis infection. The tuberculin skin test (TST) is frequently false positive in BCG vaccinated health care workers. QuantiFERON-TB GOLD (QFT-G) is a sensitive and specific interferon-gamma release assay unaffected by BCG vaccination. This study compared TST and QFT-G in the diagnosis of latent TB infection in BCG-vaccinated health care workers. 39 health care workers exposed to a smear-positive TB patient were enrolled. Initial TST was positive in 33 (84.6%) cases, but only 4 (10.2%) cases using QFT-G. TST conversion occurred in 2/6 (33.3%), compared to 4/32(12.5%), cases using QFT-G. A higher proportion of QFT converters was associated with intimate contact, although not reaching statistical significance. Face-to-face contact >1 h was significantly associated with QFT-G conversion >or=0.7 IU/ml (OR 8.63, 95%CI 1.08-69.07, p=0.04). Agreement between TST and QFT G was 18.0%, (kappa: -0.03). Concordance between TST and QFT (>or=0.35 IU/ml) conversion was 40.0%(kappa=-0.40), and 60.0%(kappa=0.00) if QFT >or=0.7 IU/ml was used. Agreement increased with increasing TST cut-offs. TST is not useful in contact investigation among BCG-vaccinated health care workers, in an area with intermediate burden of TB. QFT may provide additional information for the diagnosis and strategic management of preventive treatment of LTBI in BCG vaccinated health care workers in a country with intermediate burden of TB. PMID- 18418799 TI - BCG scar and tuberculin reactivity in children and adults. AB - Bacille Calmette-Guerin (BCG) vaccination generally leads to scar formation and tuberculin skin test (TST) reactivity. This study aimed at analysing these 2 parameters and their correlation in a setting with a low prevalence of tuberculosis. Retrospectively, we analysed 314 children and 390 adults living in Sweden and known from records or individual recall to have undergone BCG vaccination. A BCG scar was present in 161 (51%) of the children and in 340 (87%) of the adults. Among children with a scar, 94 (58%) were TST-positive (>or=6 mm) compared to 23 (15%) of 154 children lacking a visible scar. Among adults with a scar, 258 (76%) were TST- positive compared to 23 (46%) of 50 with no scar. Out of 152 non-vaccinated adults, 142 (94.4%) were TST-negative. When 175 TST negative health care students were BCG-vaccinated in a prospective part of the study, 174 (99%) were found to develop a scar. In essence, the study showed a positive correlation between scar presence and TST reactivity. Furthermore, BCG vaccination of adults in the present setting resulted in consistent scar formation, while scar prevalence in previously vaccinated children was low. PMID- 18418800 TI - Infections and antibiotic prescribing in Swedish nursing homes: a cross-sectional study. AB - The aim of this study was to present and assess the treatment of infections in Swedish nursing homes. It included 58 nursing homes with 3002 residents. During 3 months, nurses in the nursing homes recorded all infections requiring a physician's opinion. Of the 889 infectious episodes, 84% were treated with antibiotics. Many of the antibiotics were issued after indirect contact with the physician (38%). Indications for antibiotics were in 55% of the cases urinary tract infections (UTI), in 17% skin and soft-tissue infections and in 15% respiratory tract infections (RTI). The most common antibiotics were penicillins (38%), followed by quinolones (23%) and trimethoprim (18%). For the major indication, lower UTI in women, half of the cases were not treated according to the recommendations. The main concerns were length of treatment and overprescribing of quinolones. For the second major diagnosis, pneumonia, the high use of doxycycline could be questioned. Continuing education on infections and their treatment in nursing homes is needed. Training should preferably include both physicians and nurses as a high proportion of antibiotics is issued without direct contact with the physician. PMID- 18418801 TI - Ethanol lock therapy to treat tunnelled central venous catheter-associated blood stream infections: results from a prospective trial. AB - In order to assess the efficacy of 70% ethanol locks in addition to antibiotic therapy to treat tunnelled central venous catheter-associated bloodstream infections, a pilot study of 19 patients was performed prospectively using ethanol locks for 5 d in addition to antibiotic therapy to treat tunnelled central venous catheter-associated bacteraemia. 12 patients had mono-microbial infections and 7 had polymicrobial isolates. 17 of 19 patients completed ethanol lock therapy. 15 of 17 patients completing ethanol lock therapy had no recurrence of the original organism and retained their catheter for a median of 36 and an average of 47 d following initiation of ethanol lock therapy. These results demonstrate the safety and potential efficacy of this technique against a broad range of potentially virulent organisms. The intervention was acceptable to both staff and patients with no significant side-effects. These preliminary results from our prospective pilot study suggest that ethanol lock therapy is safe and easily integrated into clinical practice, and may have utility in treating central venous catheter-associated infections, avoiding removal of catheters in patients requiring long-term venous access. PMID- 18418802 TI - Candida albicans versus non-albicans bloodstream infection in patients in a tertiary hospital: an analysis of microbiological data. AB - Considerable changes in the relative frequency of systemic infections due to various Candida species as well as their in vitro susceptibility patterns have been noted in several parts of the world. We performed an analysis of microbiological data of patients with candidaemia at the University general (tertiary) hospital of Heraklion, Greece. During the study period (March 2001 to July 2006) 140 patients had candidaemia. Among them, 64/140 (46%) had candidaemia due to C. albicans and 76/140 (54%) due to non-albicans species (19/76, 25%, C. glabrata; 30/76, 40%, C. tropicalis; 20/76, 26%, C. parapsilosis; 2/76, 3%, C. lusitaniae; 3/76, 4%, C. krusei; and 2/76, 3%, C. guilliermondii). 75 isolates were tested for in vitro susceptibility to antifungal agents with E-test. No isolate was found to be resistant to amphotericin. From 34 C. albicans isolates, 5 (15%) were not susceptible to itraconazole, and 1 (3%) to fluconazole. The C. guilliermondii and the C. lusitaniae isolates were not susceptible to itraconazole. All 11 C. glabrata isolates were not susceptible to ketoconazole and itraconazole, with only 5 (45%) to fluconazole. In line with results of other relevant studies, we documented that a considerable proportion of Candida bloodstream infections were due to non-albicans Candida species. PMID- 18418803 TI - IgM and avidity of IgG antibodies in primary HHV-6 infections. AB - Primary HHV-6 infection could be diagnosed by enzyme immunoassay from a single serum using IgG avidity test based on results obtained from 43 patients, 26 with seroconversion and 17 with variable antibody levels. The avidity was less than 30% in all patients with seroconversion. HHV-6 IgM appeared non-specific. PMID- 18418804 TI - Escherichia coli: an unknown and infrequent cause of community acquired pneumonia. AB - Our aim was to describe the incidence, clinical characteristics and outcome of community acquired pneumonia (CAP) caused by Escherichia coli through the analysis of a cohort of patients with this condition. This study includes all the patients who were admitted to our hospitals because of CAP caused by E. coli, diagnosed with highly reliable microbiological techniques, such as blood culture, bronchoscopic protected specimen brush (PSB) or transthoracic needle aspiration (TNA). 29 patients were enrolled, representing 0.4% of CAP cases admitted. Main symptoms were fever and dyspnoea. 18 patients were classified into class IV and class V of the Pneumonia Severity Index (PSI). Diagnosis was based on blood culture in 24 cases, PSB in 4 cases and by TNA in 1 case. Three of the patients died, the longer time evolution of the symptoms being the only factor related to higher mortality (p<0.05). Mean hospitalization time was 7.1+/-3.1 d, and correlated with severity at admission (r=0.43; p<0.003). This study demonstrates that CAP caused by E. coli is infrequent. It has an unspecific presentation and mortality rate is 10.3%, associated with longer time before admission to hospital. PMID- 18418805 TI - Breakthrough VZV infection after immunization, presenting as herpes zoster. AB - An immunocompromized, VZV-vaccinated child had a breakthrough infection with VZV, acquired at a day-care centre during a chickenpox outbreak. Interestingly, the infection manifested as herpes zoster of 1 dermatome. Typing showed wild-type virus, which suggests that exogenous reinfection with a new strain may present as herpes zoster. PMID- 18418806 TI - Swimming performance in surf: the influence of experience. AB - This study tested the hypothesis (H1) that surf swimming involves a quantifiable experience component. Sixty-five beach lifeguards with (n = 35) and without surf experience (n = 30) completed: a best effort 200-m swim in a 25-m pool, a calm and a surf sea; an anthropometric survey; maximum effort 30-s swim bench test; 50 m pool swim (25 m underwater). In both groups, time to swim 200 m was slower in calm seas than in the pool and slower in surf than in either calm seas or the pool (p < 0.05). Calm sea swim times of the two groups did not differ significantly, but the no experience group was, on average (Sp-pooled variance), 49 s (62) slower on the 200-m swim in the surf conditions (p < 0.05). A stepwise regression identified surf experience as a predictor of surf swim time (R(2) = 0.32, p < 0.01). It is concluded that there is a significant and quantifiable (18 %) experience factor in surf swimming. This limits the usefulness of pool swim times and other land-based tests as predictors of surf swimming performance. The hypothesis (H1) is accepted. PMID- 18418807 TI - Effects of pedal type and pull-up action during cycling. AB - The aim of this study was to determine the influence of different shoe-pedal interfaces and of an active pulling-up action during the upstroke phase on the pedalling technique. Eight elite cyclists (C) and seven non-cyclists (NC) performed three different bouts at 90 rev . min (-1) and 60 % of their maximal aerobic power. They pedalled with single pedals (PED), with clipless pedals (CLIP) and with a pedal force feedback (CLIPFBACK) where subjects were asked to pull up on the pedal during the upstroke. There was no significant difference for pedalling effectiveness, net mechanical efficiency (NE) and muscular activity between PED and CLIP. When compared to CLIP, CLIPFBACK resulted in a significant increase in pedalling effectiveness during upstroke (86 % for C and 57 % NC, respectively), as well as higher biceps femoris and tibialis anterior muscle activity (p < 0.001). However, NE was significantly reduced (p < 0.008) with 9 % and 3.3 % reduction for C and NC, respectively. Consequently, shoe-pedal interface (PED vs. CLIP) did not significantly influence cycling technique during submaximal exercise. However, an active pulling-up action on the pedal during upstroke increased the pedalling effectiveness, while reducing net mechanical efficiency. PMID- 18418808 TI - Volume vs. intensity in the training of competitive swimmers. AB - The present study aimed at comparing a high-volume, low-intensity vs. low-volume, high-intensity swim training. In a randomized cross-over design, 10 competitive swimmers performed two different 4-week training periods, each followed by an identical taper week. One training period was characterized by a high-training volume (HVT) whereas high-intensity training was prevalent during the other program (HIT). Before, after two and four weeks and after the taper week subjects performed psychometric and performance testing: profile of mood states (POMS), incremental swimming test (determination of individual anaerobic threshold, IAT), 100 m and 400 m. A small significant increase in IAT was observed after taper periods compared to pre-training (+ 0.01 m/s; p = 0.01). Maximal 100-m and 400-m times were not significantly affected by training. The POMS subscore of "vigor" decreased slightly after both training periods (p = 0.06). None of the investigated parameters showed a significant interaction between test-time and training type (p > 0.13). Nearly all (83 %) subjects swam personal best times during the 3 months after each training cycle. It is concluded that, for a period of 4 weeks, high-training volumes have no advantage compared to high-intensity training of lower volume. PMID- 18418809 TI - Energy balance during backpacking. AB - Energy expenditure and energy intake were determined in three subjects during a 160-kilometer backpacking trip. Prior to the trip, heart rate and oxygen consumption during treadmill walking while carrying a backpack were measured. Subjects recorded heart rate during hiking. Heart rate was used to estimate energy expenditure based on the heart rate : oxygen consumption relationship (Heart Rate Method). Expenditure was also estimated from distance walked and elevation gained (Terrain Method). Subjects recorded food consumption, and were weighed upon finishing the hike. Mean heart rate during hiking was 105 +/- 12 beats per minute. According to the Heart Rate Method, net expenditure averaged 3410 +/- 955 kilocalories on days 1 - 4, and 2586 +/- 974 kilocalories on day 5. Net expenditure from the Terrain Method was approximately 28 % lower. Gross expenditure, using the Heart Rate Method, averaged 4928 +/- 1050 kilocalories on days 1 - 4, and 3550 +/- 1052 kilocalories on day 5. Energy intake averaged 2134 +/- 625 kilocalories on days 1 - 4, and 1117 +/- 320 kilocalories on day 5. Expected weight loss due to the caloric deficit was 1.8 +/- 0.4 kilograms, and actual weight loss was 1.7 +/- 0.6 kilograms. Subjects expended nearly 5000 kilocalories per day and did not consume sufficient food to maintain body mass. PMID- 18418810 TI - Changes of mental stress biomarkers in ultramarathon. AB - We investigated the possible influence of an exhaustive physical exercise on mental stress biomarkers (serotonin, tryptophan, and beta-endorphin) along with dopamine, noradrenaline and free fatty acids in an ultramarathon race in which 45 km was run on the first day and 90 km on the second. We obtained serum samples at 6 different time points during and after the race from 18 Japanese male runners who completed the marathon. Overall changes of serum serotonin and tryptophan concentrations were statistically significant according to ANOVA for repeated measurements (p < 0.05). Serum serotonin levels elevated rapidly on the first day with the post hoc Tukey's test. Tryptophan concentrations inversely decreased during the race, possibly because of utilization for synthesis of serotonin. Levels of beta-endorphin appeared to increase on the first and second days, but were not statistically significant. In conclusion, serum serotonin, tryptophan and beta-endorphin appeared to be used for mental stress markers in physical exercise. PMID- 18418812 TI - Chemokines and autoimmune thyroid diseases. AB - Chemokines are a family of small, structurally related molecules that regulate cell trafficking of various types of leukocytes through interactions with their seven-transmembrane, G protein-coupled receptors. Their major function is the recruitment of leukocytes to inflammation sites, but they also play roles in tumor growth, angiogenesis, organ sclerosis, and autoimmunity. A variety of evidence has accumulated to support the concept that thyroid follicular cells as well as intrathyroidal lymphocytes are able to produce CC and CXC chemokines, which, in turn, promote the initiation and maintenance of an inflammatory process resulting in autoimmune thyroid diseases (AITD). Overexpression of several chemokines in AITD has been demonstrated. Moreover, alterations of CCL2, CCL5, CXCL9, and CXCL10 have been shown in circulation of many patients with AITD. In subjects with Graves' disease, antithyroid drug treatment, radioactive iodine ablation, and thyroidectomy can significantly reduce CXCL10 levels. The measurement of chemokines in serum of AITD patients might provide a useful parameter for the evaluation and prediction of disease activity and progression. Further experimental and clinical studies will expand our understanding of the clinical implications of chemokine detection and the effects of chemokines on the pathogenesis of AITD. PMID- 18418811 TI - Dental data of the Athens 2004 Olympic and Paralympic Games. AB - The Athens University, School of Dentistry, accepted the challenge to organize the Dental Health Services in the Athens 2004 Olympic and Paralympic Games in order to provide the best quality of oral health services to the athletes, coaches, escort members and Olympic Village personnel. Data from the whole activity protocol of the Athens 2004 Games Dental Health Services - the reception, admission and treatment protocols, the facilities and the infrastructure, the number of cases treated per specialty and the experience gained - were recorded. During the Olympic Games, there were more than 1400 dental cases in more than 650 patients, elite athletes, escort members, coaches and staff of the Olympic Village. Among them 313 fillings, 100 root canal therapies, 57 mouthguards and 9 dental trauma cases were treated. During the Paralympic Games, there were more than 240 dental cases in more than 220 patients. Among them 73 fillings, 12 root canal therapies, 21 extractions and 3 dental trauma cases were treated. In such events, highly trained dentists are needed and if possible, specialized in operative dentistry or endodontics. The role of team dentist seems to be of great importance. PMID- 18418813 TI - Insulin analogues with increased mitogenic potency--are they safe? PMID- 18418814 TI - Synthesis and in vitro evaluation of alpha-GalCer epimers. AB - alpha-GalCer (also known as KRN7000) is an immunomodulatory glycolipid that is known to potently activate invariant natural killer T (NKT) cells upon CD1d mediated stimulation. Because Th1 and Th2 cytokines, which are released after alpha-GalCer presentation, antagonize each other's effects, alpha-GalCer analogues that induce a biased Th1/Th2 response are highly awaited. In this context, we report the synthesis and in vitro evaluation of alpha-Gal-D-xylo-Cer and two alpha-Gal-L-lyxo-Cer analogues, one with the natural acyl chain, the other with a truncated chain. PMID- 18418815 TI - Fluorescent naphthyl- and anthrylazoles from the catalytic coupling of phenylazoles with internal alkynes through the cleavage of multiple C-H bonds. PMID- 18418816 TI - A supramolecular catalyst for the decarboxylative hydroformylation of alpha,beta unsaturated carboxylic acids. PMID- 18418817 TI - A structure-controlled investigation of lipase enantioselectivity by a path planning approach. AB - A novel approach based on efficient path-planning algorithms was applied to investigate the influence of substrate access on Burkholderia cepacia lipase enantioselectivity. The system studied was the transesterification of 2 substituted racemic acid derivatives catalysed by B. cepacia lipase. In silico data provided by this approach showed a fair qualitative agreement with experimental results, and hence the potential of this computational method for fast screening of racemates. In addition, a collision detector algorithm used during the pathway searches enabled the rapid identification of amino acid residues hindering the displacement of substrates along the deep, narrow active site pocket of B. cepacia lipase and thus provided valuable information to guide the molecular engineering of lipase enantioselectivity. PMID- 18418818 TI - N-terminal specific fluorescence labeling of proteins through incorporation of fluorescent hydroxy acid and subsequent ester cleavage. AB - We have developed a novel method to attach a fluorescent label at the N terminus of proteins through a four-base codon-mediated incorporation of a fluorescent hydroxy acid and subsequent cleavage of the ester bond in a cell-free translation system. We found that a fluorescent-labeled p-amino-L-phenyllactic acid was successfully incorporated downstream of N-terminal tag peptides in response to a CGGG codon, and the tag peptides could be removed through ester cleavage to leave the fluorescent hydroxy acid at the N terminus of the proteins. Immunoprecipitation analysis revealed that ester cleavage occurred spontaneously during the translation reaction. The efficiency of the ester cleavage and the yield of the labeled proteins were dependent on the peptide tag sequence. We demonstrate that the insertion of an asparagine residue between the N-terminal T7 tag and the fluorescent hydroxy acid achieved both quantitative ester cleavage and efficient expression of the labeled proteins. The present method is a potential tool for N-terminal specific labeling of proteins with various compounds. PMID- 18418819 TI - Rapid and efficient DNA strand cross-linking by click chemistry. AB - Click chemistry has been used to covalently cross-link complementary DNA strands between bases to form very stable duplexes. Several alkyne- and azide-modified uracil monomers were used to evaluate the effect of the linkers on the efficiency of the click reaction. All cross-linked duplexes had much higher thermal stabilities than non-cross-linked ones, with increases in melting temperature of up to 30 degrees C. In some cases, the conversion was near-quantitative, and the reaction was complete in 5 min. PMID- 18418820 TI - Tuning colloidal interactions through coordination chemistry. AB - The present work addresses the question of the range and amplitude of bridging attraction that is induced between surfactant micelles functionalized with complexing groups in the presence of coordination centers. An alkylethoxylated ester phosphate (AEP) is synthesized from a non-ionic surfactant and anchored into DTAB micelles. In the absence of any coordination center, functionalized micelles repel each other. Phase behavior, dynamic light scattering and small angle neutrons scattering (SANS) experiments show that this repulsive interaction is switched to attractive by the addition of coordination centers such as aluminum cations. The extent of the composition range of coexisting phase depends on the concentration of coordination center and on the pH. Analysis of the structure factor obtained from SANS shows that the range of attraction is determined by the molecular dimension of the chelating surfactant, while the depth can be tuned with the concentration of coordination center and the pH. The strong influence of the pH is interpreted as arising from the condensation of aluminium cations that lead to high functional polynuclear complexes. PMID- 18418821 TI - Pressure-induced emission enhancement of a series of dicyanovinyl-substituted aromatics: pressure tuning of the molecular population with different conformations. AB - A series of dicyanovinyl-substituted aromatic compounds (Ar-DCV; Ar=9 anthracenyl, 1-naphthyl, 1-pyrenyl) with dual fluorescence are prepared, and their emission properties--when molecularly dispersed in a polymer medium--are investigated under pressure perturbation. The total emission intensity is enhanced drastically from ambient pressure up to 70 kbar. Emission 30-107 times more intense than that at ambient pressure is observed at higher pressure. In dual emission, the enhancement of the local excited state (LE state) is significantly different from that of the intramolecular charge-transfer state (ICT state). The intensity of the ICT emission increases faster (30-370 times) than that of the LE emission (less than 20 times). In accordance with spectroscopic data, emission dynamics at different pressures, and computational studies on the molecular conformations of these compounds, a kinetic model is proposed to explain the effect of pressure on the emissive properties of the Ar DCV compounds from the point of view of pressure-dependent populations of the species in the ground state. PMID- 18418823 TI - The electrochemical grafting of a mixture of substituted phenyl groups at a glassy carbon electrode surface. AB - Two-component substituted aryl groups are simultaneously grafted onto the surface of a glassy carbon electrode by electrochemical reduction of a binary mixture of two aryl diazonium salts in acetonitrile. The electrochemical deposition is achieved potentiostatically and two different mixtures with four different ratios of diazonium salts are used. The binary mixtures comprise: 1) 4-nitrophenyl diazonium and 4-bromophenyl diazonium cations and 2) 4-bromophenyl diazonium and N,N-diethylaniline diazonium cations. The chemical composition of the two component films is determined by cyclic voltammetry in an electrolyte inert for electroactive groups such as nitrophenyl and bromophenyl. X-ray photoelectron spectroscopy is also used to evaluate the surface concentration of each grafted substituted phenyl group. The surface concentration of the substituted phenyl group for which the corresponding diazonium cation is the most easily reduced is higher than its concentration in the mixture of the deposition solution. The usefulness of binary films is also discussed. PMID- 18418822 TI - (Thermo)dynamic role of receptor flexibility, entropy, and motional correlation in protein-ligand binding. AB - The binding of 2-amino-5-methylthiazole to the W191G cavity mutant of cytochrome c peroxidase is an ideal test case to investigate the entropic contribution to the binding free energy due to changes in receptor flexibility. The dynamic and thermodynamic role of receptor flexibility are studied by 50 ns-long explicit solvent molecular dynamics simulations of three separate receptor ensembles: W191G binding a K(+) ion, W191G-2a5mt complex with a closed 190-195 gating loop, and apo with an open loop. We employ a method recently proposed to estimate accurate absolute single-molecule configurational entropies and their differences for systems undergoing conformational transitions. We find that receptor flexibility plays a generally underestimated role in protein-ligand binding (thermo)dynamics and that changes of receptor motional correlation determine such large entropy contributions. PMID- 18418824 TI - Selective hydrogen oxidation in the presence of C3 hydrocarbons using perovskite oxygen reservoirs. AB - Perovskite-type oxides, ABO(3), can be successfully applied as solid "oxygen reservoirs" in redox reactions such as selective hydrogen combustion. This reaction is part of a novel process for propane oxidative dehydrogenation, wherein the lattice oxygen of the perovskite is used to combust hydrogen selectively from the dehydrogenation mixture at 550 degrees C. This gives three key advantages: it shifts the dehydrogenation equilibrium to the side of the desired products, heat is generated, thus aiding the endothermic dehydrogenation, and it simplifies product separation (H(2)O vs H(2)). Furthermore, the process is safer since it uses the catalysts' lattice oxygen instead of gaseous O(2). We screened fourteen perovskites for activity, selectivity and stability in selective hydrogen combustion. The catalytic properties depend strongly on the composition. Changing the B atom in a series of LaBO(3) perovskites shows that Mn and Co give a higher selectivity than Fe and Cr. Replacing some of the La atoms with Sr or Ca also affects the catalytic properties. Doping with Sr increases the selectivity of the LaFeO(3) perovskite, but yields a catalyst with low selectivity in the case of LaCrO(3). Conversely, doping LaCrO(3) with Ca increases the selectivity. The best results are achieved with Sr-doped LaMnO(3), with selectivities of up to 93 % and activities of around 150 mumol O m(-2). This catalyst, La(0.9)Sr(0.1)MnO(3), shows excellent stability, even after 125 redox cycles at 550 degrees C (70 h on stream). Notably, the activity per unit surface area of the perovskite catalysts is higher than that of doped cerias, the current benchmark of solid oxygen reservoirs. PMID- 18418825 TI - Specific permeability of chiral amino acids through functional molecular membranes composed of an amphiphilic graft peptide. PMID- 18418826 TI - Homolytic S-S bond dissociation of 11 bis(thiocarbonyl)disulfides R-C(=S)-S-S C(=S)R and prediction of a novel rubber vulcanization accelerator. AB - The structures and energetics of eight substituted bis(thiocarbonyl)disulfides (RCS(2))(2), their associated radicals RCS(2)(*), and their coordination compounds with a lithium cation have been studied at the G3X(MP2) level of theory for R = H, Me, F, Cl, OMe, SMe, NMe(2), and PMe(2). The effects of substituents on the dissociation of (RCS(2))(2) to RCS(2)(*) were analyzed using isodesmic stabilization reactions. Electron-donating groups with an unshared pair of electrons have a pronounced stabilization effect on both (RCS(2))(2) and RCS(2)(*). The S-S bond dissociation enthalpy of tetramethylthiuram disulfide (TMTD, R = NMe(2)) is the lowest in the above series (155 kJ mol(-1)), attributed to the particular stability of the formed Me(2)NCS(2)(*) radical. Both (RCS(2))(2) and the fragmented radicals RCS(2)(*) form stable chelate complexes with a Li(+) cation. The S-S homolytic bond cleavage in (RCS(2))(2) is facilitated by the reaction [Li(RCS(2))(2)](+)+Li(+)-->2 [Li(RCS(2))](*+). Three other substituted bis(thiocarbonyl) disulfides with the unconventional substituents R = OSF(5), Gu(1), and Gu(2) have been explored to find suitable alternative rubber vulcanization accelerators. Bis(thiocarbonyl)disulfide with a guanidine-type substituent, (Gu(1)CS(2))(2), is predicted to be an effective accelerator in sulfur vulcanization of rubber. Compared to TMTD, (Gu(1)CS(2))(2) is calculated to have a lower bond dissociation enthalpy and smaller associated barrier for the S-S homolysis. PMID- 18418827 TI - Functional macromolecules from single-walled carbon nanotubes: synthesis and photophysical properties of short single-walled carbon nanotubes functionalised with 9,10-diphenylanthracene. AB - 9,10-Diphenylanthracene (DPA), a well-studied organic chromophore (Phi(fl) = 0.98) that exhibits electroluminescence, has been covalently bound through 2 (ethylthio)ethylamido linkers to the carboxylic acid groups of short, soluble single-walled carbon nanotubes (sSWNTs) of 1 microm average length, and the resulting DPA-functionalised sSWNT (DPA- sSWNT) macromolecular adducts (4.6 wt % DPA content) characterised by solution (1)H NMR, Raman and IR spectroscopy and thermogravimetric analysis. Comparison of the quenching of DPA fluorescence (steady-state and time-resolved) and of the transient optical spectra of sSWNTs and DPA-sSWNTs show that the covalent linkage boosts the interaction between the DPA and the sSWNT units. DPA-sSWNTs exhibit emission in the near-IR region from 1100-1400 nm with an enhanced quantum yield (Phi = 5.7x10(-3)) compared with sSWNTs (Phi = 3.9x10(-3)). PMID- 18418828 TI - [Cu@Sn(9)](3-) and [Cu@Pb(9)](3-): intermetalloid clusters with endohedral Cu atoms in spherical environments. PMID- 18418829 TI - Learning from molecules in distress. PMID- 18418830 TI - Chiral phosphoric acid catalyzed enantioselective Friedel-Crafts alkylation of indoles with nitroalkenes: cooperative effect of 3 A molecular sieves. PMID- 18418831 TI - Monitoring of beet armyworm resistance to spinosad and methoxyfenozide in Mexico. AB - BACKGROUND: Resistance to spinosad and methoxyfenozide has been studied in several insect pests, but there is a lack of information on Spodoptera exigua (Hubner) in Mexico. Therefore, evidence for the development of resistance in this pest to both compounds was examined. The effects of methoxyfenozide on reproductive parameters of S. exigua adults were also determined. RESULTS: Third instars from a field population were exposed for 24 h to the LC(50) of spinosad or methoxyfenozide for over six generations (G(2)-G(7)). No significant reduction in susceptibility to either compound was detected for up to five generations. In G(7), LC(50) values for insects exposed to spinosad and methoxyfenozide were respectively 2.75-fold and 1.25-fold greater than for G(1) larvae. Oral treatment with methoxyfenozide reduced the fecundity and fertility of G(7) adults, confirming sublethal effects on reproduction. Finally, five populations (Se-La Floriza, Se-Lazareto, Se-Bachigualato, Se-Los Agustinos and Se-Villa de Arista) of S. exigua were collected from fields in three states of Mexico for resistance monitoring to spinosad and methoxyfenozide. With the exception of Se-Villa de Arista, the other populations showed significant resistance to spinosad, with resistance ratios between 16- and 37-fold, compared with a susceptible laboratory colony. In contrast, only one population (Se-Lazareto) showed significant resistance to methoxyfenozide (13-fold). CONCLUSION: Resistance management programmes should be established, particularly in areas where S. exigua has developed resistance to spinosad. PMID- 18418832 TI - Synthesis of somatostatin mimetics based on 1-deoxynojirimycin. AB - The synthesis of novel somatostatin mimetics from 1-deoxynojirimycin (DNJ) is described. The dipeptide mimetic, which respectively displayed the side chains of tryptophan and lysine at the nitrogen and O6 atoms of the iminosugar scaffold is a ligand (K(i)=3.2 microM) for the human somatostatin receptor subtype 4 (hSSTR4) but has lower affinity (K(i)>100 microM) for hSSTR5. A benzylated analogue of the Trp-Lys mimetic displays higher affinity for hSSTR5 (K(i)=5 microM). PMID- 18418833 TI - Exploring acyclic nucleoside analogues as inhibitors of Mycobacterium tuberculosis thymidylate kinase. AB - In the search for novel inhibitors of the enzyme thymidine monophosphate kinase of Mycobacterium tuberculosis (TMPKmt), an attractive target for novel antituberculosis agents, we report herein the discovery of the first acyclic nucleoside analogues that potently and selectively inhibit TMPKmt. The most potent compounds in this series are (Z)-butenylthymines carrying a naphtholactam or naphthosultam moiety at position 4, which display K(i) values of 0.42 and 0.27 microM, respectively. Docking studies followed by molecular dynamics simulations performed to rationalize the interaction of this new family of inhibitors with the target enzyme revealed a key interaction between the distal substituent and Arg 95 in the target enzyme. The fact that these inhibitors are more easily synthesizable than previously identified TMPKmt inhibitors, together with their potency against the target enzyme, makes them attractive lead compounds for further optimization. PMID- 18418834 TI - Tuning the magnetic properties of Li(x)CrTi0.25Se2 (0.03 < or = x < or = 0.7) by directed deintercalation of lithium. AB - X-ray diffraction (XRD), in situ energy-dispersive X-ray diffraction (EDXRD), X ray absorption near-edge structure (XANES), extended X-ray absorption fine structure (EXAFS), and magnetic measurements were applied to investigate the effects of lithium deintercalation on pseudolayered Li(0.70)CrTi(0.25)Se(2). A detailed picture of structural changes during the deintercalation process was obtained by combining the results of EDXRD and EXAFS. Removal of Li from the host guest complex leads to anisotropic contraction of the unit cell with stronger impact on the c axis, which is the stacking axis of the layers. The EDXRD experiments evidence that the shrinkage of the lattice parameters with decreasing x(Li) in Li(x)CrTi(0.25)Se(2) is nonlinear in the beginning and then becomes linear. Analysis of the EXAFS spectra clearly shows that the Cr/Ti-Se distances are affected in a different manner by Li removal. The Cr-Se bond lengths decrease, whereas the Ti-Se bonds lengthen when the Li content is reduced, which is consistent with XRD data. Magnetic measurements reveal a change from predominantly antiferromagnetic exchange (theta(p) = -300 K) interactions for the pristine material to ferromagnetic exchange interactions (theta = 25 K) for the fully intercalated material. Thus, the magnetic properties can be altered under ambient conditions by directed adjustment of the dominant magnetic exchange. The unusual magnetic behavior can be explained on the basis of the variation of the metal-metal distances and the Cr-Se-Cr angles with x, which were determined by Rietveld refinements. Owing to competing ferromagnetic and antiferromagnetic exchange interactions and disorder, the magnetic ground state of the intercalated materials is characterized by spin-glass or spin-glass-like behavior. PMID- 18418835 TI - Application of MCD spectroscopy and TD-DFT to nonplanar core-modified tetrabenzoporphyrins: effect of reduced symmetry on nonplanar porphyrinoids. AB - The optical spectra of a series of core-modified tetrabenzoporphyrins were analyzed to determine the effects of core modification, ligand folding, and partial benzo substitution at the ligand periphery on the electronic structure by using magnetic circular dichroism (MCD) and NMR spectroscopy, X-ray crystallography, cyclic and differential pulse voltammetry, and TD-DFT calculations. Planar 21-carba-, 21-thia-, 21,23-dithia-, and 21-oxa-23 thiatetrabenzo[b,g,l,q]porphyrins reported previously were studied together with the previously unreported 21-oxa- and 21-carba-23 thiatetrabenzo[b,g,l,q]porphyrins. The optical properties of these compounds are compared to those of tetrabenzo[b,g,l,q]-, 5,10,15,20-tetraphenyl-, 5,10,15,20 tetraphenyltetrabenzo[b,g,l,q]-21-thia-, 5,10,15,20-tetraphenyltetrabenzodithia-, 5,10,15,20-tetraphenyldibenzo[g,q]-21,23-dithia-, 5,10,15,20 tetraphenyldibenzo[b,l]-21,23-dithia-, 5,10,15,20-tetraphenyltribenzo[g,q,l]-21 thia-, and 5,10,15,20-tetraphenylbenzo[b]-21-thiaporphyrins. Michl's perimeter model and Gouterman's four-orbital model are used to conceptualize the results and to account for red shifts commonly observed in the spectral bands of nonplanar porphyrinoids. PMID- 18418836 TI - Metal-mediated formation of carbon-halogen bonds. AB - Organic halides represent basic starting materials for numerous metal-catalyzed organic transformations. Generally, the carbon-halogen is broken in the first step, that is, an oxidative addition reaction, of the catalytic cycle. On the other hand, very little is known about the reverse reaction, carbon-halogen reductive elimination from a transition-metal center. In this Concept article, we describe the examples of C(sp(3))-halide and C(sp(2))-halide reductive elimination reactions which demonstrate that this type of reactivity can be quite common in organometallic chemistry. Although the thermodynamic driving force for the formation of carbon-halogen bonds is relatively small, the kinetic barrier for these reactions can also be low. Thus, C-halide reductive elimination can compete favorably with the more established organic transformations, such as C-C reductive elimination. PMID- 18418837 TI - Diverse reactions of 1,4-dilithio-1,3-dienes with nitriles: facile access to tricyclic Delta1-bipyrrolines, multiply substituted pyridines, siloles, and (Z,Z) dienylsilanes by tuning of substituents on the butadienyl skeleton. AB - Addition cyclization of 1,2,3,4-tetrasubstituted 1,4-dilithio-1,3-dienes (Type I) with four equivalents of various aromatic nitriles in the presence of hexamethylphosphoramide (HMPA) gives exclusively fully substituted pyridines in moderate to good yields. Similarly, trisubstituted pyridines can be prepared by the reaction of 2,3-dialkyl- or diaryl-substituted 1,4-dilithio-1,3-dienes (Type II) with nitriles. However, five- or six-membered-ring fused 2,3-disubstituted 1,4-dilithio-1,3-dienes (Type III) reacted with various aromatic and aliphatic nitriles without alpha-hydrogen atoms to afford tricyclic Delta1-bipyrrolines in high yields. The reaction of six-membered-ring fused 2,3-disubstituted 1,4 dilithio-1,3-diene (Type III) with 2-cyanopyridine afforded the corresponding pyridine, and no tricyclic Delta1-bipyrroline was observed. Seven-membered-ring fused dilithiodienes reacted with PhCN or trimethylacetonitrile to afford the corresponding pyridines in good yield. When 1,2,3,4-tetrasubstituted dilithio reagents (Type I) were treated with Me3SiCN, a tandem silylation/intramolecular substitution process readily occurred to yield siloles, whereas the reaction of 2,3-disubstituted dilithio reagents (Types II and III) with Me3SiCN gave rise to (Z,Z)-dienylsilanes with high stereoselectivity. These results revealed that the formation of tricyclic Delta1-bipyrrolines, pyridines, siloles, and (Z,Z) dienylsilanes are strongly dependent on the substitution patterns of the dilithio butadienes and the nature of the nitriles employed. PMID- 18418838 TI - Mixed-transition-metal acetylides: synthesis and characterization of complexes with up to six different transition metals connected by carbon-rich bridging units. AB - The synthesis and reaction chemistry of heteromultimetallic transition-metal complexes by linking diverse metal-complex building blocks with multifunctional carbon-rich alkynyl-, benzene-, and bipyridyl-based bridging units is discussed. In context with this background, the preparation of [1-{(eta(2)-dppf)(eta(5) C(5)H(5))RuC[triple bond]C}-3-{(tBu(2)bpy)(CO)(3)ReC[triple bond]C}-5 (PPh(2))C(6)H(3)] (10) (dppf = 1,1'-bis(diphenylphosphino)ferrocene; tBu(2)bpy = 4,4'-di-tert-butyl-2,2'-bipyridyl; Ph = phenyl) is described; this complex can react further, leading to the successful synthesis of heterometallic complexes of higher nuclearity. Heterotetrametallic transition-metal compounds were formed when 10 was reacted with [{(eta(5)-C(5)Me(5))RhCl(2)}(2)] (18), [(Et(2)S)(2)PtCl(2)] (20) or [(tht)AuC[triple bond]C-bpy] (24) (Me = methyl; Et = ethyl; tht = tetrahydrothiophene; bpy = 2,2'-bipyridyl-5-yl). Complexes [1 {(eta(2)-dppf)(eta(5)-C(5)H(5))RuC[triple bond]C}-3-{(tBu(2)bpy)(CO)(3)ReC[triple bond]C}-5-{PPh(2)RhCl(2)(eta(5)-C(5)Me(5))}C(6)H(3)] (19), [{1-[(eta(2) dppf)(eta(5)-C(5)H(5))RuC[triple bond]C]-3-[(tBu(2)bpy)(CO)(3)ReC[triple bond]C] 5-(PPh(2))C(6)H(3)}(2)PtCl(2)] (21), and [1-{(eta(2)-dppf)(eta(5) C(5)H(5))RuC[triple bond]C}-3-{(tBu(2)bpy)(CO)(3)ReC[triple bond]C}-5 {PPh(2)AuC[triple bond]C-bpy}C(6)H(3)] (25) were thereby obtained in good yield. After a prolonged time in solution, complex 25 undergoes a transmetallation reaction to produce [(tBu(2)bpy)(CO)(3)ReC[triple bond]C-bpy] (26). Moreover, the bipyridyl building block in 25 allowed the synthesis of Fe-Ru-Re-Au-Mo- (28) and Fe-Ru-Re-Au-Cu-Ti-based (30) assemblies on addition of [(nbd)Mo(CO)(4)] (27), (nbd = 1,5-norbornadiene), or [{[Ti](mu-sigma,pi-C[triple bond]CSiMe(3))(2)}Cu(N[triple bond]CMe)][PF(6)] (29) ([Ti] = (eta(5) C(5)H(4)SiMe(3))(2)Ti) to 25. The identities of 5, 6, 8, 10-12, 14-16, 19, 21, 25, 26, 28, and 30 have been confirmed by elemental analysis and IR, (1)H, (13)C{(1)H}, and (31)P{(1)H} NMR spectroscopy. From selected samples ESI-TOF mass spectra were measured. The solid-state structures of 8, 12, 19 and 26 were additionally solved by single-crystal X-ray structure analysis, confirming the structural assignment made from spectroscopy. PMID- 18418839 TI - Clusterization, electrophoretic deposition, and photoelectrochemical properties of fullerene-functionalized carbon nanotube composites. AB - We have successfully developed a new methodology for the self-organization of C(60) molecules on the sidewall of carbon nanotubes for use in photoelectrochemical devices. Novel nanocarbon composites of fullerene (e.g., C(60)) and highly soluble, chemically functionalized single-walled carbon nanotubes (f-SWNT) have been prepared by the rapid injection of a poor solvent (e.g., acetonitrile) into a mixed solution of C(60) and f-SWNT in o dichlorobenzene. Measurements by using scanning electron microscopy of cast samples revealed that the composites are categorized into three groups; i) f-SWNT bundles covered with layers of C(60) molecules, ii) round, large C(60) clusters (sizes of 500-1000 nm) containing f-SWNT, and iii) typical, round C(60) clusters (sizes of 150-250 nm). The electrophoretic deposition of the composites onto a nanostructured SnO(2) electrode yielded the hierarchical film with a gradient composition depending on the difference in the mobilities of C(60) and f-SWNT during the electrophoretic process. The composite film exhibited an incident photon-to-photocurrent efficiency as high as 18 % at lambda=400 nm under an applied potential of 0.05 V vs. SCE. The photocurrent generation efficiency is the highest value among carbon nanotube-based photoelectrochemical devices in which carbon nanotubes are deposited electrophoretically, electrostatically or covalently onto semiconducting electrodes. The highly aligned structure of C(60) molecules on f-SWNT can rationalize the efficient photocurrent generation. The results obtained here will provide valuable information on the design of carbon nanotube-based molecular devices. PMID- 18418841 TI - Fullerene C60-perylene-3,4:9,10-bis(dicarboximide) light-harvesting dyads: spacer length and bay-substituent effects on intramolecular singlet and triplet energy transfer. AB - Novel covalent fullerene C(60)-perylene-3,4:9,10-bis(dicarboximide) (C(60)-PDI) dyads (1-4) were synthesized and characterized. Their electrochemical and photophysical properties were investigated. Electrochemical studies show that the reduction potential of PDI can be tuned relative to C(60) by molecular engineering through altering the substituents on the PDI bay region. It was demonstrated using steady-state and time-resolved spectroscopy that a quantitative, photoinduced energy transfer takes place from the PDI moiety, acting as a light-harvesting antenna, to the C(60) unit, playing the role of energy acceptor. The bay-substitution (tetrachloro [1 and 2] or tetra-tert butylphenoxy [3 and 4]) of the PDI antenna and the linkage length (C(2) [1 and 3] or C(5) [2 and 4]) to the C(60) acceptor are important parameters in the kinetics of energy transfer. Femtosecond transient absorption spectroscopy indicates singlet-singlet energy-transfer times (from the PDI to the C(60) unit) of 0.4 and 5 ps (1), 4.5 and 27 ps (2), 0.8 and 12 ps (3), and 7 and 50 ps (4), these values being ascribed to two different conformers for each C(60)-PDI system. Subsequent triplet-triplet energy-transfer times (from the C(60) unit to the PDI) are slower and in the order of 0.8 ns (1), 6.2 ns (2), 2.7 ns (3), and 9 ns (4). Nanosecond transient absorption spectroscopy of final PDI triplet states show a marked influence of the bay substitution (tetrachloro- or tetra-tert-butylphenoxy), and triplet-state lifetimes (10-20 micros) and the PDI triplet quantum yields (0.75 0.52) were estimated. The spectroscopy showed no substantial solvent effect upon comparing toluene (non-polar) to benzonitrile (polar), indicating that no electron transfer is occurring in these systems. PMID- 18418840 TI - Size- and structure-selective noncovalent recognition of saccharides by tetraethyl and tetraphenyl resorcinarenes in the gas phase. AB - The noncovalent complexation of tetraethyl and tetraphenyl resorcinarenes with mono-, di-, and oligosaccharides was studied with negative-polarization electrospray ionization quadrupole ion trap and electrospray ionization Fourier transform ion cyclotron resonance mass-spectrometric analysis. The saccharides formed 1:1 complexes with deprotonated resorcinarenes, which exhibited clear size and structure selectivity in their complexation. In the case of the monosaccharides, hexoses formed much more abundant and kinetically stable complexes than pentoses or deoxyhexoses. A comparison of the mono-, di-, and oligosaccharides revealed that both the relative abundance and stability of the complexes increase up to biose and triose, but start to decrease after that point, as the length of the oligosaccharide is increased. This behavior was rationalized by comparing the lowest-energy conformations of the complexes formed between the resorcinarene and oligosaccharides. This comparison was achieved by using theoretical calculations and X-ray crystal studies. PMID- 18418842 TI - Nephrotoxicity induced by N-(3,5-dichlorophenyl)-3-hydroxysuccinamic acid in male and female Fischer 344 rats. AB - The agricultural fungicide N-(3,5-dichlorophenyl)succinimide (NDPS) is a more potent nephrotoxicant in female rats than in males. Similarly, nephrotoxicant NDPS metabolites studied to date in male and female rats have also demonstrated gender differences, being twice as potent as nephrotoxicants in females as in males. The purpose of this study was to examine the nephrotoxic potential of N (3,5-dichlorophenyl)-3-hydroxysuccinimide (3-NDHSA) in male and female Fisher 344 rats to determine if gender differences in nephrotoxic potential also exist for this metabolite. Rats (four per group) were administered a single intraperitoneal (i.p.) injection of 3-NDHSA (0.1, 0.2 or 0.4 mmol kg(-1)) or vehicle, and renal function was monitored at 24 and 48 h. 3-NDHSA 0.1 mmol kg(-1) did not induce nephrotoxicity in male or female rats. In male rats, 3-NDHSA 0.2 mmol kg(-1) induced mild nephrotoxicity seen as diuresis and transient, mild proteinuria. However, 3-NDHSA 0.4 mmol kg(-1) induced marked nephrotoxicity. In female rats, 3 NDHSA 0.2 mmol kg(-1) induced mild nephrotoxicity, as evidenced by transient diuresis and proteinuria. As in males, 3-NDHSA 0.4 mmol kg(-1) induced marked nephrotoxicity. These results indicate that, unlike NDPS and other nephrotoxic NDPS metabolites, 3-NDHSA does not exhibit gender differences in nephrotoxic potential. In addition, in comparison with NDPS and other nephrotoxic NDPS metabolites, 3-NDHSA is a less potent nephrotoxicant that NDHS or 2-NDHSA and similar to NDPS in nephrotoxic potential in male rats. PMID- 18418843 TI - Cytochrome c: a non-invasive biomarker of drug-induced liver injury. AB - Limitations of existing biomarkers to detect liver injury in experimental animals highlight the need for additional tools to predict human toxicity. The utility of cytochrome c (cyt c) as a biomarker in serum and urine was evaluated in two rodent liver injury models. Adult Sprague-Dawley rats treated with acetaminophen or D-galactosamine (GalN) showed dose- and time-dependent histomorphological changes and TUNEL staining in liver consistent with hepatocellular necrosis, apoptosis and inflammation up to 72 h. Matching changes in serum alanine transaminase (ALT), aspartate transaminase (AST) and cyt c peaked at 24 h for either drug at the highest dose, cyt c falling rapidly at 48 hours with ALT and AST remained high. Intracellular transit of cyt c from mitochondria to the cytoplasm in damaged hepatocytes, and then to peripheral circulation, was observed by immunohistochemistry. Correlation coefficients between cyt c and serum diagnostic tests indicate the liver to be the primary source of cyt c. Urinary analysis for cyt c revealed time-dependent increase at 6 h, peaking at 24 h in GalN-treated rats in contrast with irregular patterns of urinary ALT and AST activity. Histological changes detected at 6 h preceded altered ALT, AST and cyt c at 12 and 18 h, respectively, in GalN-treated rats. These studies demonstrate cyt c to be a useful indicator of hepatic injury in rodents and support its utility as a non-invasive predictor of drug-induced hepatotoxicity, when utilized as a potential urinary biomarker. PMID- 18418844 TI - Lack of cross-tolerance following heat and cadmium exposure in functional MDCK monolayers. AB - Exposure of monolayers of Madin-Darby canine kidney epithelial (MDCK) cells to a mild heat stimulus induces a state of physiological thermotolerance in which epithelial barrier function is maintained following a second more severe heat stress. We have previously shown that expression of exogenous HSP70 fully mimics the effects of the conditioning heat stress. Exposure of MDCK cells to elevated temperatures or medium containing CdCl2 caused a robust increase in cellular levels of HSP70. Pretreatment of MDCK monolayers with cadmium but not heat caused a small protection of epithelial barrier function against a second challenge with cadmium. In addition, a prior exposure of monolayers to cadmium at levels sufficient to induce HSP70 expression and increased cellular chaperone activity did not afford protection against a subsequent thermal challenge. Therefore multiple stress-specific cellular pathways impinge on the ability of heat shock proteins to induce physiological thermotolerance. Occludin, a component of tight junctions, is induced in MDCK cells engineered to express high levels of exogenous HSP70, potentially accounting for an elevation in baseline resistance. However neither basal levels of occludin, nor alterations in occludin expression, were correlated with epithelial barrier function in MDCK cells either exposed to elevated temperatures or challenged with cadmium. PMID- 18418845 TI - Biochemical and enzymatic changes after black scorpion Heterometrus fastigiousus Couzijn envenomation in experimental albino mice. AB - The toxic effects of Asian black scorpion Heterometrus fastigiousus (Family, Scorpionidae) venom were determined in albino mice (NIH strain). Venom was isolated and fractioned by Sepharose CL-6B column chromatography. The toxicity of fractioned venom was determined in albino mice by subcutaneous envenomation. The LD(50) of venom was found to be 15 mg kg(-1) body weight and range of molecular weight of venom proteins responsible for toxicity was found from 9.5-63 kDs. The effects of fractioned venom on different biochemical and enzymatic parameters in blood serum and gastrocnemius muscle tissue of albino mice were determined after experimental envenomation. An increase in serum levels of glucose, free amino acids, uric acid, pyruvic acid and total protein was observed while a decrease in the cholesterol level in serum was observed after 4 h of envenomation. Increase in alkaline phosphatase (ALP), acid phosphatase (ACP), lactic dehydrogenase (LDH) and glutamate-pyruvate transaminase (GPT) enzyme activity in serum was observed. Glycogen content in liver, atria, ventricle, rectus abdominus and gastrocnemius muscle was decreased after experimental envenomation. Activity of ALP, ACP, LDH, GPT, AChE and Na+K+ATPase enzymes in gastrocnemius muscle tissue of envenomed albino mice was studied. Inhibition in ALP, AChE and Na+K+ATPase enzyme activity and increase in ACP, LDH and GPT enzyme activity was observed in gastrocnemius muscle after scorpion envenomation. In vitro studies with AChE and Na+K+ATPase enzymes indicated that enzymatic activity of AChE was inhibited competitively by fractioned venom in gastrocnemius muscle. PMID- 18418847 TI - A pilot study of self-monitoring urine flow in people with long-term urinary catheters. AB - Self-monitoring of urine flow may be beneficial in preventing catheter-related problems. An intervention was pilot tested using a single group design with 11 individuals over 6 months. Feasibility of the intervention, performance of new measures (awareness, self-monitoring, and self- management of urine flow), and health outcomes were assessed. The intervention was well received by participants, none of whom withdrew voluntarily from the study. All data were collected at intake, and bimonthly at 2, 4, and 6 months. Nine of 11 participants reported that the intervention had helped them to pay attention to fluid intake. Episodes of UTI decreased over the course of the intervention, with the greatest drop between 2 and 4 months. PMID- 18418846 TI - Laminin terminates the Netrin/DCC mediated attraction of vagal sensory axons. AB - Vagal sensory axons navigate to specific sites in the bowel during fetal life. Netrin/deleted in colorectal cancer (DCC) were found to mediate the attraction of vagal sensory axons to the fetal mouse gut. We tested the hypothesis that laminin 111 can reverse the chemoattractive effects of netrin and act as a stop signal for vagal sensory axons. Laminin-111-expressing cells were located in the E12 and E16 mouse bowel by in situ hybridization. At E12, these cells extended centrifugally from the endoderm; by E16, laminin-111 expressing cells were found in the mucosa and outer gut mesenchyme. A similar pattern was seen in preparations of E13 and E15 mouse gut labeled with antibodies to laminin. Application of DiI to nodose ganglia identified vagal sensory axons growing into the fetal bowel. These terminals were found to avoid concentrations of laminin or to terminate at laminin-delimited boundaries. Soluble laminin inhibited the preferential growth of nodose neurites toward netrin-secreting cells (p < 0.01). This effect was mimicked by a peptide, YIGSR, a sequence within the beta1 chain of laminin-111 (p < 0.004) and antagonized by a peptide, IKVAV, a sequence within the alpha1 chain of laminin-111. Antibodies to beta1-integrins were also able to reverse the inhibitive effects of laminin and restore the attraction of nodose neurites towards netrin-1-secreting cells. Soluble laminin inhibited the preferential growth of nodose neurites toward a cocultured explant of foregut. These findings suggest that laminin terminates the attraction of vagal sensory axons towards sources of netrin in the developing bowel. PMID- 18418848 TI - Direct resequencing of the complete ERBB2 coding sequence reveals an absence of activating mutations in ERBB2 amplified breast cancer. AB - Gene amplification is among the most common genetic abnormalities that cause cancer. One of the most clinically important gene amplifications in human cancer causes extensive reduplication of ERBB2. A variety of cancers also occasionally harbor somatic mutations in ERBB2. Gene amplification and activating mutations both have predictive value for clinical response to targeted inhibitors. Since the number of gene copies in an amplicon may exceed 100, and since amplicons may encompass multiple genes, high-resolution analysis of gene amplifications poses considerable technical challenges. We have overcome this obstacle by using emulsion-based resequencing to determine the sequence of many independently amplified individual DNA molecules in parallel. We used this high throughput sequencing technology to analyze ERBB2 mutational status in five ERBB2 amplified cell lines (four breast, one ovarian) and two breast tumors. Genomic DNA was isolated and the 28 exons of ERBB2 were independently amplified. Amplicons were then pooled at equimolar ratios, subjected to emulsion PCR (emPCR) and finally to picotiter plate pyrosequencing. High-quality sequence data were obtained for all amplicons analyzed and no activating mutations within ERBB2 were identified. Although we did not find activating mutations within the multiple copies of ERBB2 in these samples, the results establish the utility of this technology as a feasible and cost-effective approach for high resolution analysis of amplified genes. PMID- 18418849 TI - Cytologic findings of acquired cystic disease-associated renal cell carcinoma: a report of two cases. AB - Renal tumors may arise in the setting of end-stage renal cell disease. The risk is 100 times that of the normal population with an incidence ranging from 3-7%. The most common malignant tumor to arise in the setting of acquired cystic disease of the kidney is the acquired cystic disease-associated renal cell carcinoma (ACD-associated RCC). The cytomorphologic features of ACD-associated RCC, which has not been described previously, show moderately cellular specimens composed of clusters of cells with papillary configuration. The cells ranged from polygonal to columnar and contained abundant eosinophilic granular cytoplasm. The nuclei were round and centrally located, and the chromatin was finely granular with prominent central nucleoli that corresponded to Fuhrman's grade 3 nucleolar size. The main differential diagnosis is type 2 papillary renal cell carcinoma, from which it can be distinguished based on clinical findings only at this moment. PMID- 18418850 TI - Pericardial effusion as primary manifestation of metastatic cutaneous adenoid cystic carcinoma: diagnostic cytopathology from an exfoliative sample. AB - Adenoid cystic carcinoma (ACC) occurs not only as a tumor of salivary glands, but also in very unusual locations, such as in the skin. Only very few cases of primary cutaneous of ACC have metastasized to the lymph nodes and lungs. We present a 53-year-old man with metastasis of the pericardium from a primary cutaneous ACC (PCACC) of the scalp, which had been surgically treated 14 years ago. Exfoliative cytologic findings from pericardial effusion included small clusters of basaloid cells with occasional cystlike spaces containing mucoid material. To our knowledge, this is the first case of pericardial metastasis from a PCACC. PMID- 18418851 TI - A strategy for efficient handling of fresh tumor needle biopsies that allows histological and cytopathological assessment. AB - Neo-adjuvant chemotherapy prior to surgery is used in the management of many pediatric solid tumors, and diagnosis is therefore valuable and is frequently made by percutaneous needle biopsy. We describe a method that enhances tissue preservation and obtains a sample for rapid cytopathological assessment. Biopsies are placed in Ham's F(10) culture's medium in theatre and transferred to pathology. The biopsies are retrieved from the medium and dealt as before (submit to cytogenetics; fix in glutharaldheyde; snap frozen at -80 degrees C and routine histology). An equal amount of 90% alcohol is then added to the Ham culture's medium fluid received from theatre before performing a cytospin preparation and a cell clot. We used this method in the diagnosis of 16 tumors demonstrating that this allows a more efficient handling of the biopsy, makes possible a same day diagnosis, enhances the quality of the immunohistochemistry and maximizes the amount of tissue available for diagnosis. PMID- 18418852 TI - Evaluation of performance of EUS-FNA in preoperative lymph node staging of cancers of esophagus, lung, and pancreas. AB - We reviewed the cytologic and histologic diagnoses and EUS report of 77 consecutive patients who had undergone EUS-FNA preoperative staging for esophageal, lung, and pancreatic cancers at our institution. A total of 122 EUS FNA lymph nodes were identified. Thirty of 77 cases had histologic follow-up. Using surgical node staging and/or surgical resection as the reference standard, the sensitivity, specificity, accuracy, and positive and negative predictive values were 75%, 95%, 89%, 86%, and 90%, respectively, for EUS-FNA node staging. We compared cytologically malignant and benign lymph node groups with eight EUS parameters including the total number of lymph nodes found by EUS, the shape, margin, long axis, short axis, echogenicity, location of the lymph node, and EUS tumor staging. We found that the short axis is the best EUS feature to predict malignancy. Lymph nodes found in an abdominal location in esophageal and lung cancer are likely malignant. PMID- 18418853 TI - Adult granulosa cell tumor of the ovary: fine-needle-aspiration cytology of 10 cases and review of literature. AB - Adult granulosa cell tumor (GCT) of the ovary is mostly diagnosed in postmenopausal women. They typically secrete estrogen, which stimulates the endometrium to proliferate and cause abnormal bleeding. This study reviews the cytologic features of adult GCT of the ovary diagnosed by fine-needle aspiration (FNA). We reviewed slides from ten cases diagnosed by CT guided FNA from 1995 to 2007 at our institutions. Smears were stained with Diff-Quik and Papanicolaou stains. Patient's history and histologic diagnosis were also available and reviewed for all cases. The patients ranged in age from 39 to 83 yr. All 10 cases were hypercellular with both large and small overlapping cell clusters and individual cells. The cytologic features identified included: naked nuclei (10/10 cases), Call-Exner bodies (7/10 cases), blood vessels with prominent perivascular tumor cell growth (4/10 cases), spindle-shaped hyperchromatic stromal cells within cellular clusters (6/10 cases), mixed inflammation (3/10 cases), tumor cell necrosis (1/10 cases), and prominent metachromatic stroma seen in association with blood vessels (1/10 cases). Moderate to scant delicate cytoplasm was also seen (10/10 cases). Small, punctuate cytoplasmic vacuoles were also noted (7/10 cases) and were occasionally prominent (3/10 cases). In general nuclear to cytoplasmic ratios were high although lower than those typically seen in a lymphoma or small-cell carcinoma. Nuclei were generally centrally located although eccentrically located nuclei were consistently seen in a minority of cells. Nuclei were monotonous in size showing slightly convoluted (occasional rentiform and fetiform nuclei) to polygonal outlines. Prominent, central nucleoli were also seen (4/10 cases). Nuclear grooves were also seen (9/10 cases). No atypical mitotic activity was identified in any of the 10 cases (0/10 cases). In summary, the above cytologic features can also help in the cytologic diagnosis of adult GCTs. PMID- 18418854 TI - Cytopathologist-performed ultrasound-guided fine-needle aspiration and core needle biopsy: a prospective study of 500 consecutive cases. AB - In experienced hands, palpation-guided fine-needle aspiration is an excellent diagnostic tool. However, there is a movement toward using imaging guidance to target all masses. This study examines the feasibility of cytopathologist performed ultrasound-guided FNA (UG-FNA) and ultrasound-guided core-needle biopsy (UG-CNB) of masses in the breast, thyroid, head and neck, and other superficial sites. Between January 8, 2007 and June 28, 2007, 415 consecutive patients with 500 masses that were (1) nonpalpable (2) palpable but sonographically heterogeneous requiring ultrasound guidance to target the abnormal areas, or (3) palpable but located near a structure to be avoided underwent cytopathologist performed UG-FNA and/or UG-CNB. There were 395 (79%) nonpalpable masses and 105 (21%) palpable masses. The nondiagnostic rate was 1.4%. All cases undergoing both FNA and CNB were diagnostic. With continuing medical education in radiology and ultrasound-guided procedures, training on phantoms, and practice on palpable masses before transition to nonpalpable masses, the author was able to successfully perform UG-FNA/CNB of nonpalpable masses and targeted UG-FNA/CNB of palpable masses. In the hands of the cytopathologist, ultrasound guidance offers a new tool to revitalize and redefine the role of the interventional cytopathologist. A new era in cytopathology and fine-needle aspiration may be on the horizon. PMID- 18418855 TI - Hamartoma of the lung. PMID- 18418856 TI - Cyst fluid cytology of cerebellar hemangioblastoma: a case report. AB - Hemangioblastoma, an uncommon central nervous system neoplasm, commonly located in the cerebellum and more rarely in the spinal cord. In this report, we described the cytologic features of a cerebellar hemangioblastoma in a 53-year old man, present in cystic fluid obtained at cranioctomy. Cytologic preparations were characterized by well preserved, scattered dissociate, and occasional groups of stromal cells with round to oval nuclei, fine chromatin, and lacelike or granular, vacuolated eosinophilic cytoplasm with a foamy appearances. In addition, calcific material, endothelial-like cells, blood, and macrophages were seen. The radiologic impression, cytologic features, and location led us to suggest the possibility of hemangioblastoma. Subsequent histopathological material revealed the characteristic features of hemangioblastoma. PMID- 18418857 TI - Fine-needle aspiration cytology of Sister Mary Joseph's (paraumbilical) nodules. AB - During a period of 5 years (2001-2005), six cases of Sister Mary Joseph's nodule (umbilical metastasis) were diagnosed by fine needle aspiration cytology (FNAC). In all the cases, FNAC of umbilical nodules was the first investigation and subsequently the patients were investigated for the primary tumor. The primary carcinoma was found in the stomach in three cases, ovary in two cases and one case was of non-Hodgkin's lymphoma. Based on our findings, we suggest FNAC as an initial diagnostic procedure in cases suspected of umbilical metastasis. It is not only simple, fast, accurate and inexpensive but can also save the patient from other invasive diagnostic procedures. PMID- 18418858 TI - Insulin-like growth factor I improves aspects of mycophenolate mofetil-impaired anastomotic healing in an experimental model. AB - BACKGROUND: Patients taking immunosuppressants after transplantation may require intestinal surgery. Mycophenolate mofetil (MMF) has been found to impair the healing of colonic anastomoses in rats. This study examined whether insulin-like growth factor (IGF) I prevents MMF impairment of anastomotic healing. METHODS: Sixty-three rats were divided into three groups (MMF, MMF/IGF and control). Animals underwent a sigmoid colon anastomosis with a 6/0 suture, and were killed on days 2, 4 and 6 after surgery. Investigations included bursting pressure measurement, morphometric analysis, and assessment of mucosal proliferation by 5 bromo-2'-deoxyuridine and Ki67 immunohistochemistry of the anastomoses. RESULTS: The leak rate was three of 21, one of 20 and two of 20 in the MMF, MMF/IGF-I and control groups respectively. Anastomotic bursting pressures were significantly lower in the MMF group than in the control group on days 2 and 4, but there was no significant difference by day 6. Values in the MMF/IGF-I and control groups were similar. Colonic crypt depth was significantly reduced in MMF-treated animals on days 2 and 4, but this impairment was attenuated by IGF-I on day 4. Similarly, IGF-I reduced the negative impact of MMF on mucosal proliferation on days 2 and 6. CONCLUSION: Exogenous IGF-I improves some aspects of MMF-impaired anastomotic healing. PMID- 18418859 TI - Outcomes of colorectal anastomoses during pelvic exenteration for gynaecological malignancy. AB - BACKGROUND: Although pelvic exenteration is frequently indicated during surgery for gynaecological malignancy, performing a colorectal anastomosis remains contentious because of concern about leakage. This study evaluated the safety of performing a low colorectal anastomosis during pelvic exenteration for gynaecological malignancy. METHODS: Between April 2001 and December 2006, 145 consecutive patients underwent low colorectal anastomosis without (122) or with (23) a stoma after pelvic exenteration for advanced primary or recurrent gynaecological malignancy. Subjects were assessed in terms of five patient-, four disease- and two surgery-related variables. The proportion of patients with each risk factor for leakage was found, and the rate of symptomatic anastomotic leakage was determined. RESULTS: The mean age of the patients was 53.5 (range 10 77) years and the most common diagnosis was ovarian cancer (77.9 per cent). The mean operating time was 453 (range 145-845) min and the mean blood loss was 1080 (range 110-10 500) ml; 95 patients (65.5 per cent) required a blood transfusion. Of the 145 patients, 81 (55.9 per cent) had patient-related, 94 (64.8 per cent) had disease-related and 67 (46.2 per cent) had surgery-related variables associated with a risk of leakage. Symptomatic anastomotic leakage developed in three patients (2.1 per cent). CONCLUSION: Although patients with gynaecological malignancy carry considerable risks associated with anastomotic leakage, carefully executed low colorectal anastomosis during pelvic exenteration was found to be safe. PMID- 18418860 TI - APC mutation spectrum in ileoanal pouch polyps resembles that of colorectal polyps. AB - BACKGROUND: Ileoanal pouch polyps commonly develop following restorative proctocolectomy in patients with familial adenomatous polyposis (FAP). In FAP adenomas, the relationship between germline and somatic adenomatous polyposis coli (APC) mutations is determined by 'just right' beta-catenin signalling in tumour cells, with respect to the 20-amino acid beta-catenin-binding/degradation repeats (20AARs) in the APC protein. However, the relationship varies, with upper gastrointestinal polyps typically retaining three to four 20AARs and colonic polyps retaining one or two. The aim of this study was to establish the mutational spectrum in ileoanal pouch polyps, to ascertain whether polyp development resembled that typical of small or large bowel. METHODS: Some 151 pouch adenomas were screened from 46 patients with known germline APC mutations for 'second hits' acquired through loss of heterozygosity and truncating mutations. The number of 20AARs remaining after the 'second hit' was calculated. RESULTS: Loss of heterozygosity was rare in pouch polyps except when the germline mutation left one 20AAR. Overall, the combined alleles left two to three 20AARs in 40 of 51 polyps with an identified 'second hit'. This was significantly fewer than in upper gastrointestinal polyps, and more than in colorectal adenomas. CONCLUSION: Tissue environment appears to influence the position of the 'second hit' in pouch polyps and the mutations resemble those of large bowel polyps. PMID- 18418861 TI - Impact of body mass index and tobacco smoking on outcome after open appendicectomy. AB - BACKGROUND: The effect of body mass index (BMI) and smoking on the risk of perforated appendix and postoperative complications in patients undergoing open appendicectomy for acute appendicitis was studied. METHODS: Record linkage was used to identify 6676 male construction workers who underwent open appendicectomy for acute appendicitis between 1971 and 2004. Multivariable binomial logistic regression analyses were performed. RESULTS: After adjustment for age, calendar period and BMI, smoking was significantly associated with an increased risk of perforated appendicitis (PA) (P = 0.004). The relative risk was 1.29 (95 per cent confidence interval 1.11 to 1.50) among current smokers with more than 10 pack years of tobacco use. In patients with non-perforated appendicitis (NPA), the relative risk of overall postoperative complications was significantly associated with BMI (P < 0.001), and was 2.60 (1.71 to 3.95) in obese patients and 1.51 (1.03 to 2.22) in current smokers with more than 10 pack-years of tobacco use. In patients with PA, overweight, obesity and smoking status were not associated with an increased risk of overall postoperative complications. CONCLUSION: Perforation due to acute appendicitis was associated with current tobacco smoking. A BMI of 27.5 kg/m(2) or more and current smoking were associated with overall postoperative complications in patients with NPA. PMID- 18418862 TI - Immunoglobulin G4-related sclerosing cholangitis in patients resected for presumed malignant bile duct strictures. AB - BACKGROUND: Immunoglobulin (Ig) G(4)-related lymphoplasmacytic sclerosing pancreatitis has been described in the context of autoimmune pancreatitis mimicking distal cholangiocarcinoma. The aim of this study was to assess the occurrence of this entity in benign bile duct strictures in patients resected for presumed hilar cholangiocarcinoma. METHODS: Of 185 patients who had undergone resection of proximal bile ducts on suspicion of hilar cholangiocarcinoma between January 1984 and June 2005, 32 (17.3 per cent) had a benign bile duct stricture on histopathological examination. After re-evaluation, further immunohistochemical analysis was performed on specimens from patients with features of autoimmune-like disease. RESULTS: The periductal stroma in 15 patients showed features of autoimmune-like disease (diffuse, moderate to severe lymphoplasmacytic infiltration with marked fibrosis). Abundant IgG(4)-positive plasma cell infiltration around the bile duct lesions was seen in two of these. Although not significant, patients with features of autoimmune-like disease on histological changes showed a higher incidence of recurrent biliary complications than those without (P = 0.250). CONCLUSION: Features of autoimmune-like bile duct disease were seen in almost half (15 of 32) of patients with benign hilar strictures resected for presumed hilar cholangiocarcinoma. Frank IgG(4)-related sclerosing disease was found in only two of the 15 patients with autoimmune-like bile duct disease. PMID- 18418864 TI - Human germline and somatic cells have similar TP53 and Kirsten-RAS gene single base mutation frequencies. AB - Understanding the risk of offspring inheriting rare mutations, and the frequencies at which these mutations are present in germ cells can be explored with direct analysis of human semen samples. The present work utilized the ultrasensitive PCR/RE/LCR mutation assay to detect, identify and determine the prevalence single base substitution mutations in the TP53 and KRAS genes in human sperm. Four disease-associated base sites in the TP53 and KRAS genes, three of which are known to be heritable to live, term offspring, were studied in sperm from eleven human semen specimens. Eight of the specimens (73%) displayed single base substitution mutations, and 30% of all base sites tested were found to harbor mutations ranging in prevalence from 1 x 10(-6) to 1 x 10(-5) wild type sperm. These germ cell single base substitution mutation frequencies are very similar to somatic tissue TP53 and KRAS mutation frequencies. Equivalent single base mutation frequencies in both germ and somatic cells suggest that there is no unusual selection or mutation protective process operating premeiotically in the germline, and that a selection bias at the level of sperm viability, conception, early cleavage, implantation, and/or embryogenesis operates to exclude the majority of these TP53 mutations and all of the activating KRAS mutations. PMID- 18418863 TI - Proteomic biomarkers for prenatal bisphenol A-exposure in mouse immune organs. AB - Many investigators have encountered difficulty in clarifying the risks of exposure to bisphenol A (BPA), an endocrine disrupting chemical in epidemiological studies or animal experiments. In the present study, we developed biomarkers of BPA-induced proteomic alterations in immune organs of mouse offspring that were prenatally exposed to BPA (15 and 300 mg/L of drinking water; they were exposed to 8.9 +/- 1.8 mg of BPA/kg/day and 171.1 +/- 16.8 mg of BPA/kg/day, respectively) that were evaluated in terms of sex, age, and BPA exposure levels. We performed 2D-gel analyses of samples from various tissues (thymus and spleen), exposure levels, sex, and ages (3- and 7-week-old) (N = 48), and found seven proteins that were altered in a BPA dose-dependent manner. Among them, we further studiedapo-AI, DPPIII, and VAT1, which are suspected to be associated with endocrine disorders. By performing Western blots, we confirmed BPA upregulation of all three proteins. Moreover, the apo-AI mRNA levels were increased in a BPA dose-dependent manner in 3- and 7-week-old female mice. Females and young offspring were somewhat more sensitive to protein alterations than others. Our study, which is based on proteome analyses, suggests that apo AI, DPPIII, and VAT represent protein biomarkers for BPA and provide useful mechanistic clues for BPA-induced endocrine disruption. PMID- 18418865 TI - Protective effect of the cruciferous vegetable mustard leaf (Brassica campestris) against in vivo chromosomal damage and oxidative stress induced by gamma radiation and genotoxic chemicals. AB - We evaluated the possible protective effect of the popular Indian cruciferous vegetable mustard leaf (Brassica campestris) against chromosomal damage and oxidative stress induced by gamma-radiation, cyclophosphamide (CPH) and urethane (URE), in mice. In vivo bone marrow micronucleus test was performed to assess chromosomal damage, and oxidative stress was monitored by estimating the changes in lipid peroxidation and the status of glutathione (GSH) as well as redox cycle antioxidants. Pretreatment with 50-250 mg/kg body wt of mustard leaf extract (MLE) for seven days significantly reduced the frequencies of micronuclei induced by gamma-radiation, CPH and URE. The protective effect against chromosomal damage was associated with modulation of lipid peroxidation as well as an increase in GSH and the GSH-dependent enzyme glutathione S-transferase (GST). Mass spectral analysis showed the presence of glucosinolates in MLE used for the pretreatment of mice. These findings indicate that intake of the green leafy cruciferous vegetable mustard leaf can lead to protection against in vivo genotoxicity and oxidative stress. PMID- 18418866 TI - Genotoxic effects of p,p'-DDT (1,1,1-trichloro-2,2-bis-(chlorophenyl)ethane) and its metabolites in Zebra mussel (D. polymorpha) by SCGE assay and micronucleus test. AB - This is the first study to evaluate the potential genotoxicity of p,p'-DDT (1,1,1 trichloro-2,2-bis-(chlorophenyl)ethane) and its metabolites (p,p'-DDD and p,p' DDE) in the sentinel mollusc Zebra mussel (Dreissena polymorpha). DNA damage was measured using the single cell gel electrophoresis (SCGE) assay and the micronucleus test (MN test), which represent two of the more sensitive biomarkers for genotoxicity evaluation. Three different concentrations (0.1, 2, and 10 mug/L) of each compound were administered in water for 168 hr, maintaining mussels at constant laboratory conditions and collecting several specimens every 48 hr for biochemical analyses. At the same time, the bioaccumulation process and the concentration/effect relationship were checked by GC-MS/MS analyses of mussel soft tissues. The SCGE assay results showed a clear and significant (P < 0.05) relationship between DNA injuries and tested doses for all the homologues throughout the 7-day exposure period. The final DNA damage due to p,p'-DDE was almost double that of the other two homologues that showed the same toxicity pattern. The micronucleus frequency analysis confirmed the genotoxicity potential of the three homologues and p,p'-DDE showed the highest irreversible DNA damage. The capability of Zebra mussels to biotransform the administered compound in the other homologues was demonstrated by multiple regression analyses carried out between the MN frequencies and the concentrations of the different homologues in the mussel soft tissues. A greater genotoxic potential of the p,p'-DDE with respect to the other two chemicals was revealed. PMID- 18418867 TI - Inhibition of the 5' to 3' exonuclease activity of hEXO1 by 8-oxoguanine. AB - The mismatch repair pathway is responsible for maintaining genomic stability by correcting base-base mismatches and insertion/deletion loops that arise mainly via replication errors. Additionally, the mismatch repair pathway performs a central role in the cellular response to both alkylation and reactive oxygen species induced DNA damage. An important step in mismatch processing is the recruitment of hEXO1, a 5' to 3' exonuclease, by hMSH2-hMSH6 to remove the nascent DNA strand. However, very little is currently known about the capacity of hEXO1 to exonucleolytically process damaged DNA bases. Therefore, we examined whether hEXO1 can degrade double-stranded DNA substrates containing alkylated or oxidized nucleotides. Our results demonstrated that hEXO1 is capable of degrading duplex DNA containing an O6-methylguanine (O6-meG) adduct paired with either a C or a T. Additionally, the hMSH2-hMSH6 complex stimulated hEXO1 exonuclease activity on the O6-meG/T and O6-meG/C DNA substrates. In contrast, hEXO1 exonuclease activity was significantly blocked by the presence of an 8-oxoguanine adduct in both single and double stranded DNA substrates. Further, hMSH2-hMSH6 was not able to alleviate the nucleolytic block caused by the 8-oxoguanine adduct in heteroduplex DNA. PMID- 18418868 TI - Titanium dioxide nanoparticles trigger p53-mediated damage response in peripheral blood lymphocytes. AB - Titanium dioxide nanoparticles (nano-TiO2) are widely used as a photocatalyst in air and water remediation. These nanoparticles are known to induce toxicity; however, their cytotoxic mechanism is not fully understood. In this study, we investigated the underlying mechanism of nano-TiO2-induced cytotoxicity in peripheral blood lymphocytes. We examined the genotoxic effects of nano-TiO2 in lymphocytes using alkaline single-cell gel electrophoresis (Comet) and cytokinesis-block micronucleus (CBMN) assays. Lymphocytes treated with nano-TiO2 showed significantly increased micronucleus formation and DNA breakage. Western blot analysis to identify proteins involved in the p53-mediated response to DNA damage revealed the accumulation of p53 and activation of DNA damage checkpoint kinases in nano-TiO2-treated lymphocytes. However, p21 and bax, downstream targets of p53, were not affected, indicating that nano-TiO2 does not stimulate transactivational activity of p53. The generation of reactive oxygen species (ROS) in nano-TiO2-treated cells was also observed, andN-acetylcysteine (NAC) supplementation inhibited the level of nano-TiO2-induced DNA damage. Given that ROS-induced DNA damage leads to p53 activation in the DNA damage response, our results suggest that nano-TiO2 induces ROS generation in lymphocytes, thereby activating p53-mediated DNA damage checkpoint signals. PMID- 18418869 TI - Improved Comet assay for the assessment of UV genotoxicity in Mediterranean sea urchin eggs. AB - Gametes and embryos of broadcast spawners are exposed to a wide range of chemical and physical stressors which may alone, or in conjunction, have serious consequences on reproductive outcomes. In this study, two Mediterranean echinoid species, Paracentrotus lividus and Sphaerechinus granularis, were chosen as models to study the genotoxicity of UV radiation (UVR) on the eggs of broadcast spawning marine invertebrates. The single cell gel electrophoresis, or Comet assay, was successfully adapted to assess DNA strand breakage in sea urchin eggs. The results demonstrated that the genetic material of sea urchin eggs is susceptible to environmentally realistic UV exposure. The induction of DNA damage in the irradiated unfertilized eggs suggests that the previously described defense mechanisms in sea urchin eggs do not completely protect the egg's DNA against UV toxicity. Taken together, our results suggest that UV-impairment of the genetic integrity of the eggs might have a role in postfertilization failures and abnormal embryonic development. Although both species were vulnerable to UVR, embryonic development was less dramatically impaired in P.Lividus. This observation supports the postulation that species inhabiting shallower environments possess more efficient mechanisms to overcome UV-induced DNA alterations. The present demonstration of the utility and sensitivity of the Comet assay to evaluate DNA integrity in eggs from marine invertebrates opens new perspectives for monitoring the long-term effects of environmental exposure on populations and for the routine screening of substances for genotoxicity in marine systems. PMID- 18418870 TI - Cytogenetic damage in female Pakistani agricultural workers exposed to pesticides. AB - Bhawalpur is a major cotton-growing area in Pakistan. Cotton picking in Pakistan is carried out by females and as a result of the intensive use of pesticides during the growing season these females are exposed to pesticide residues in the picking season. In the present study, peripheral blood was obtained from 69 cotton pickers and 69 unexposed females and used to assess the effect of pesticide exposure on genetic damage as well as on hepatic enzymes and serum cholinesterase. The subjects were of similar average age in workers and control groups (37.55 +/- 12.75 vs. 37.52 +/- 13.47, P > 0.05). Average exposure time of the picker females was 10.26 +/- 6.14 years. Subjects from the exposed group did not use any protective measures during their work activities. Levels of serum cholinesterase were lower, and levels of alkaline phosphatase, alanine aminotransferase, and aspartate aminotransferase were higher in the exposed workers as compared with the control group (P < 0.001). The exposed group exhibited significantly increased frequencies of binucleated cells with micronuclei (12.72 +/- 3.48 vs. 4.35 +/- 2.44, P < 0.001) and total number of micronuclei in binucleated lymphocytes (16.51 +/- 4.27 vs. 5.86 +/- 3.09, P < 0.001) in comparison with subjects of the control group. The binucleated cells with micronuclei frequency also seemed to increase with age in both the groups, however, the magnitude of increase was greater in exposed group than the control. Results from the present study indicate that occupational exposure to pesticide mixtures results in cytogenetic damage in exposed females. PMID- 18418871 TI - Effects of model organophosphorous pesticides on DNA damage and proliferation of HepG2 cells. AB - Organophosphorous compounds (OPs) are commonly used pesticides. The primary mechanism of OP toxicity is the inhibition of acetylcholine esterase in the nervous system leading to a variety of acute and chronic effects. Recent studies have revealed several other targets of OPs that disturb noncholinergic biological systems. We investigated whether low concentrations of model OPs-methyl parathion (PT), methyl paraoxon (PO), and dimefox (DF)-induce DNA damage and/or affect cell proliferation in human hepatoma HepG2 cells. Genotoxicity of OPs was evaluated using the comet assay. The effect on cell proliferation was tested using the MTT assay and proliferation marker Ki-67 immunocytochemistry. The effects of OPs on mRNA expression of the DNA damage responsivegenes p53, p21, GADD45alpha, and MDM2 were determined using qRT-PCR. PT induced DNA damage at lower concentrations (1 microg/mL) than PO (100 microg/mL), whereas DF did not induce DNA damage. PT and PO caused a reduction of cell proliferation at their highest concentrations (100 microg/mL), while DF increased cell proliferation at all concentrations used (0.01-100 microg/mL). PT and PO upregulated expression of DNA damage responsive genes, while DF upregulated expression of p53, downregulated expression of p21, and had no effect on the expression of MDM2 and GADD45alpha. We conclude that PT and PO are genotoxic, while DF shows mitogenic activity. An important finding of this study is that PT had higher genotoxic potential than PO, which warrants for further investigations to correctly evaluate the hazards of exposure to these chemicals. PMID- 18418872 TI - Assessment of DNA strand breaks induced by bleomycin in barley by the comet assay. AB - Comet assay was applied to study induction and repair of DNA damage produced by bleomycin in barley genome. Experimental conditions were adapted to achieve efficient detection of both DNA single- and double-strand breaks. Substantial increase of the parameter "% of DNA in tail" was observed coupled with almost linear dependence from bleomycin concentration, more pronounced for the induction of DNA double-strand breaks. Data obtained at different recovery periods displayed rapid restoration of breakage, revealing that efficient mechanisms for repair of strand discontinuities induced by bleomycin are functional in barley DNA loop domains. PMID- 18418873 TI - Studies of thioguanine-resistant lymphocytes induced by in vivo irradiation of mice. AB - The frequency of Hprt-deficient lymphocytes in mice after in vivo gamma irradiation, has been found to vary as a function of time elapsed after exposure and irradiation dose. The frequency of mutant lymphocytes in spleen was determined using an in vitro, clonogenic assay for thioguanine-resistant T lymphocytes. Mice were exposed to single doses of 0-400 cGy from cesium-137 or to eight daily doses of 50 cGy. The time to maximum-induced mutant frequency was 3 weeks. The dose response was strikingly curvilinear at 3-5 weeks after irradiation, but less precisely defined for 10-53 weeks after exposure, being fit by either linear or quadratic dependence. Three weeks after eight daily 50 cGy exposures, mutant frequency was elevated above controls and mice exposed to 50 cGy (which were not distinct from the nonirradiated controls), but only 17% in that of mice given a single 400 cGy fraction. This fractionation effect and the curvilinearity of the early dose-response curve suggested that saturation of repair increased the yield of mutations at higher acute doses. The decline of spleen mutant frequency in mice observed between 5 and 10 weeks after irradiation may reflect selection against some mutants. The marked variation of mutant frequency, as a function of time after irradiation and of dose rate, emphasize the need to evaluate these variables carefully and consistently in future studies. PMID- 18418874 TI - D2R DNA transfer into the nucleus accumbens attenuates cocaine self administration in rats. AB - Dopamine (DA) D2 receptor (D2R) agonists and antagonists can modulate self administration behavior, conditioned place preference, and locomotor responses to cocaine. Low levels of D2R have also been observed in cocaine addicted subjects and in non human primates after chronic cocaine exposures. Prior studies had shown that D2R upregulation in the nucleus accumbens (NAc) in rodents trained to self-administer alcohol markedly attenuated alcohol preference and intake. Here we assess the effects of D2R upregulation in the NAc on cocaine intake in rats trained to self-administer cocaine. Following 2 weeks of i.v. cocaine self administration (CSA), rats were stereotaxically treated with an adenovirus that carried the D2R gene to upregulate D2R in the NAc. D2R vector treatment resulted in a significant decrease (75%) in cocaine infusions and lever presses (70%) for cocaine. This effect lasted 6 days before cocaine consumption returned to baseline levels, which corresponds roughly to the time it takes D2R to return to baseline levels. These findings show that CSA and D2R in the NAc are negatively correlated and suggest that cocaine intake is modulated in part by D2R levels in NAc. Thus strategies aimed at increasing D2R expression in NAc may be beneficial in treating cocaine abuse and addiction. PMID- 18418876 TI - The rescue of developing avian motoneurons from programmed cell death by a selective inhibitor of the fetal muscle-specific nicotinic acetylcholine receptor. AB - In an attempt to determine whether the rescue of developing motoneurons (MNS) from programmed cell death (PCD) in the chick embryo following reductions in neuromuscular function involves muscle or neuronal nicotinic acetylcholine receptors (nAChRs), we have employed a novel cone snail toxin alphaA-OIVA that acts selectively to antagonize the embryonic/fetal form of muscle nAChRs. The results demonstrate that alphaA-OIVA is nearly as effective as curare or alpha bungarotoxin (alpha-BTX) in reducing neuromuscular function and is equally effective in increasing MN survival and intramuscular axon branching. Together with previous reports, we also provide evidence consistent with a transition between the embryonic/fetal form to the adult form of muscle nAChRs in chicken that involves the loss of the gamma subunit in the adult receptor. We conclude that selective inhibition of the embryonic/fetal form of the chicken muscle nAChR is sufficient to rescue MNs from PCD without any involvement of neuronal nAChRs. PMID- 18418875 TI - Sex differences in brain developing in the presence or absence of gonads. AB - Brain sexual differentiation results from the interaction of genetic and hormonal influences. This study used a unique agonadal mouse model to determine relative contributions of genetic and gonadal hormone influences in the differentiation of selected brain regions. SF-1 knockout (SF-1 KO) mice are born without gonads and adrenal glands and are not exposed to endogenous sex steroids during fetal/neonatal development. Consequently, male and female SF-1 KO mice are born with female external genitalia and if left on their own, die shortly after birth due to adrenal insufficiency. In this study, SF-1 KO mice were rescued by neonatal adrenal transplantation to examine their brain morphology in adult life. To determine potential brain loci that might mediate functional sex differences, we examined the area and distribution of immunoreactive calbindin and neuronal nitric oxide synthase in the preoptic area (POA) and ventromedial nucleus of the hypothalamus, two areas previously reported to be sexually dimorphic in the mammalian brain. A sex difference in the positioning of cells containing immunoreactive calbindin in a group within the POA was clearly gonad dependent based on the elimination of the sex difference in SF-1 KO mice. Several other differences in the area of ventromedial hypothalamus and in POA were maintained in male and female SF-1 KO mice, suggesting gonad-independent genetic influences on sexually dimorphic brain development. PMID- 18418877 TI - Normal adult climbing fiber monoinnervation of cerebellar Purkinje cells in mice lacking MHC class I molecules. AB - Some immune system proteins have recently been implicated in the development and plasticity of neuronal connections. Notably, proteins of the major histocompatibility complex 1 (MHC class 1) have been shown to be involved in synaptic plasticity in the hippocampus and the development of projection patterns in the visual system. We examined the possible role for the MHC class 1 proteins in one well-characterized example of synaptic exuberance and subsequent refinement, the climbing fiber (CF) to Purkinje cell (PC) synapse. Cerebella from adult mice deficient for two MHC genes, H2-D1 and H2-K1, and for beta2 microglobulin gene were examined for evidence of deficient elimination of supernumerary CF synapses on their PCs. Electrophysiological and morphological evidence showed that, despite the absence of these MHC class 1 molecules, adult PCs in these transgenic mice are monoinnervated as in wild-type animals. These findings indicate that, at the level of restriction of afferent number at this synapse, functional MHC class 1 proteins are not required. PMID- 18418879 TI - The induction of cytochrome P450 1A1 by sudan dyes. AB - Azo dyes form a major class of chemically related compounds that are ubiquitous in foods, paints, printing inks, cosmetics, and also used as biological stains in histological and histopathological laboratories and clinics. Sudan I, sudan III, and sudan IV have been classified as category 3 carcinogens by International Agency for Research on Cancer. In this study, we investigated the difference between these three sudan dyes in induction of CYP1A1. We intraperitoneally treated Wistar rats with each of the three sudan dyes (I, III, and IV) for 3 days. Treatment of Wistar rats with sudan I produced the highest induction of CYP1A1 protein and mRNA whereas treatment of Wistar rats with sudan III produced about two third of CYP1A1 protein and mRNA than induced by sudan I. Furthermore, treatment of Wistar rats with sudan IV produced the lowest induction of CYP1A1 protein and mRNA which is about two third of that induced with sudan III treatment. We further investigated the effect of these sudan dyes on CYP1A1 transcription through investigating the xenobiotic response element (XRE) reporter activity in HepG2. The XRE reporter activity study showed the same trend of activity of sudan dyes comparable to the effects on CYP1A1 mRNA and protein. Immunohistochemical study revealed a differential pattern of distribution of CYP1A1 protein in rat liver among the three sudan dyes, apparent in the centrilobular and midzonal region with sudan III, progressing to panlobular with sudan I, whereas sudan IV showed a reversal of pattern of induction with the most intense staining in the periportal region. Our results suggest that there is an inverse relationship between the molecular size of the three sudan dyes and their ability to induce CYP1A1. PMID- 18418880 TI - Diagnostic accuracy of fine-needle aspiration cytology of palpable breast masses: an SROC curve with fixed and random effects linear meta-regression models. AB - We used various meta-analytic methods to compare 25 studies describing fine needle aspiration (FNA) cytologic analyses performed from 1984 to 2007 on palpable breast masses. We found that in the 25 studies examined, the sensitivity ranged from 78% to 100%, the specificity ranged from 76% to 100%, and the diagnostic odds ratio (DOR) ranged from 15.83 to 33 198. The overall diagnostic accuracy was found to be as follows: 0.93 (95% CI: 0.92-0.94) for sensitivity, 0.98 (95% CI: 0.97-0.98) for specificity, and 505.209 (95% CI: 273.08-934.95) for the DOR. The overall diagnostic accuracy according to the results of summary receiving operating characteristic (SROC) curve analysis was 0.95 +/- 0.0032, and the overall weighted area under the (receiving operating characteristic [ROC]) curve (AUC) was 0.99 +/- 0.0014. The DOR values did not show a large variation in the various positivity threshold values. However, the results of those studies had some heterogeneity. The four covariates that were added to the standard SROC model to evaluate variations in the results of the studies were the year of publication, the number of aspirations, the percentage of insufficient material, and the study design. The relative diagnostic accuracy of studies performed after 1990 was 3.98 times higher than that of studies performed before 1990. The relative DOR (RDOR) value was also found to be statistically significant (95% CI: 1.22-13.02). That result may be attributed to the technologic improvements in diagnostic tools over the years. Although it was not statistically significant, an increase in the number of aspirations caused an increase in the RDOR of the FNA cytology (95% CI: 0.52-8.11). In contrast, increasing the percent of insufficient material caused a statistically insignificant but clinically significant decrease in the RDOR of FNA (RDOR = 0.79, 95% CI: 0.21-2.98). In conclusion, our meta-analysis has shown that FNA cytologic analysis of palpable breast masses is highly accurate in the diagnostic differentiation of benign from malignant tumors. PMID- 18418881 TI - Immunocytochemical staining of p16(ink4a) protein as an adjunct test in equivocal liquid-based cytology. AB - For cervical cancer screening, HPV-DNA test is expensive and is not easily available in all clinical situations. Thus, we investigated the role of p16(ink4a) immunostaining as another adjunct test to diagnose cervical neoplasia in equivocal liquid based cytology. Eighty-seven patients were randomly selected for this study (3 patients with normal, 84 patients with abnormal including 24 ASCUS, 30 LSIL, and 30 HSIL). We performed p16(ink4a) immunostaining on ThinPrep slide and on each case from the corresponding cervical biopsy tissues. High-risk HPV-DNA testing was also performed on all the subjects. We found that the immunoreactivity of p16(ink4a) is strongly correlated with the grade of cytologic and histologic diagnoses as well as with Hybrid Capture 2. In comparing the p16(ink4a) immunostaining with the Hybrid Capture 2 for accuracy of the diagnosis of CIN II/III or a higher-grade disease in the case of ASCUS/LSIL on ThinPrep, no significant differences were observed. Our data implies that p16(ink4a) immunocytochemical staining in liquid-based cytology specimens might be used as a good adjunct test to predict cervical histology in equivocal ThinPrep tests. PMID- 18418882 TI - Fine-needle aspiration of fibrous dysplasia of bone: a worthwhile endeavor or not? AB - Fine-needle aspiration (FNA) is a minimally invasive technique which is enjoying ever-increasing popularity in the initial diagnosis of many pathologic processes. However, FNA diagnosis of neoplasms occurring within bones is less commonly employed and is not the preferred method in some types of bony lesions. Fibrous dysplasia is a primary neoplasm of bone for which it is not yet clear whether FNA can reliably yield adequate diagnostic material. Review of data from 82 cases of fibrous dysplasia diagnosed between 1990 and 2006 yielded six cases, in which diagnosis was initially attempted by FNA prior to open biopsy and surgical resection. Corresponding cytologic, histologic, and imaging characteristics of the cases were reviewed. Of the six cases in which initial diagnosis was attempted by FNA, only two of six (33%) yielded adequate diagnostic material. Smears of aspirated material in all cases contained nonspecific elements, including fragments of benign host bone and cartilage, bland stromal cells, adipocytes, blood, and debris. Importantly, the two positive FNA cases were dependent on the concurrent core needle biopsy (all smears of aspirate material were nondiagnostic). Even with image guidance, FNA is insufficient to obtain diagnostic material for cases of suspected fibrous dysplasia of bone. Core needle biopsy at least is recommended to obtain adequate material, and to reduce the risk of misdiagnosis due to sampling error. PMID- 18418883 TI - Cytology of metastatic thymic well-differentiated neuroendocrine carcinoma (thymic carcinoid) in pleural fluid: report of a case. AB - Thymic carcinoid tumors (well-differentiated neuroendocrine carcinomas) are uncommon anterior mediastinal neoplasms. These tumors are frequently accompanied by other endocrinopathies as part of a multiple endocrine neoplasia type I syndrome (MEN type I) and by paraneoplastic Cushing's syndrome and have a poor prognosis. We present the case of a 24-year-old man who presented for follow-up of thymic carcinoid with extensive bony metastases. He had recently completed radiotherapy to lesions involving his skull and mandible. An ultrasound-guided left-sided diagnostic and therapeutic thoracentesis was performed yielding 1 l of cloudy yellow fluid. The cytologic fluid preparations consisted of large "cannonballs" and atypical cell groups with salt and pepper nuclear chromatin. A panel of immunohistochemical stains were performed on the cell block material, and the atypical cells were positive for cytokeratin, synaptophysin, and chromogranin, but not for TTF1. These findings were consistent with metastatic well-differentiated neuroendocrine carcinoma (carcinoid tumor). This is the first reported case of a carcinoid tumor manifesting as large, spherical, smoothly contoured cell aggregates ("cannonballs") in a pleural fluid. Despite its rarity, a metastatic carcinoid tumor should be considered when "cannonballs" are found in effusions. PMID- 18418884 TI - Koilocytes in urinary cytology in a patient with kidney transplant. AB - We are reporting the case of a 17-year-old girl with kidney transplant under immunosuppressive treatment. Evidences of transplant malfunction led to urinary cytology to rule out BKV infection. The smears showed the presence of koilocytes. Gynecologic examination revealed numerous condylomatous lesions in the vulva, vagina, and cervix. PAP smears showed cells with moderate to severe koilocytic dysplasia. PCR performed on material retrieved from both the smears showed HPV18 DNA sequences. Koilocytes have rarely been documented in urinary cytologic examination. Since post-transplant immunosuppressed patients are prone to develop florid and extensive HPV infections, urinary cytology may prove useful for routine search of cells with this virus cytopathic effect. PMID- 18418885 TI - Gardnerella vaginalis and Lactobacillus sp in liquid-based cervical samples in healthy and disturbed vaginal flora using cultivation-independent methods. AB - Our objective was to determine the morphotype of the adherent bacteria in liquid based cytology (LBC) in smears with healthy and disturbed vaginal flora. And to use PCR technology on the same fixed cell sample to establish DNA patterns of the 16S RNA genes of the bacteria in the sample. Thirty samples were randomly selected from a large group of cervical cell samples suspended in a commercial coagulant fixative "(BoonFix)." PCR was used to amplify DNA of five bacterial species: Lactobacillus acidophilus, Lactobacillus crispatus, Lactobacillus jensenii, Gardnerella vaginalis, and Mycoplasma hominis. The LBC slides were then analyzed by light microscopy to estimate bacterial adhesion. DNA of lactobacilli was detected in all cell samples. Seventeen smears showed colonization with Gardnerella vaginalis (range 2.6 x 10(2)-3.0 x 10(5) bacteria/mul BoonFix sample). Two cases were identified as dysbacteriotic with high DNA values for Gardnerella vaginalis and low values for Lactobacillus crispatus. The sample with the highest concentration for Gardnerella vaginalis showed an unequivocal Gardnerella infection. This study indicates that the adherence pattern of a disturbed flora in liquid-based cervical samples can be identified unequivocally, and that these samples are suitable for quantitative PCR analysis. This cultivation independent method reveals a strong inverse relationship between Gardnerella vaginalis and Lactobacillus crispatus in dysbacteriosis and unequivocal Gardnerella infection. PMID- 18418886 TI - Inositol supplementation in pregnancies at risk of apparently folate-resistant NTDs. PMID- 18418887 TI - Molecular effects of lithium exposure during mouse and chick gastrulation and subsequent valve dysmorphogenesis. AB - BACKGROUND: Lithium (Li) has been associated with cardiac teratogenicity in the developing fetus. We took advantage of the association of therapeutic administration of Li with an increase in heart defects to gain insight into both normal and pathological heart and valve development with GSK-3 inhibition. The objective of this study was to define whether Li mimicry of canonical Wnt/beta catenin signaling induces cardiac valve defects. METHODS: Li was administered by a single intraperitoneal injection to the pregnant mouse on embryonic day E6.75, much earlier than heretofore analyzed. On E15.5 developing heart defects were defined by Doppler ultrasound. The embryonic hearts were analyzed for changes in patterning of active canonical Wnt expression and nuclear factor of the activated T cells-c1 (NFATc1), both key regulators of valve development. Li-exposed chick embryos were used to define the early cell populations during gastrulation that are susceptible to GSK-3 inhibition and may relate to valve formation. RESULTS: Li exposure during gastrulation decreased the number of prechordal plate (PP) cells that reached the anterior intestinal portal, a region associated with valve development. Li decreased expression of Hex, an endoderm cardiac inducing molecule, normally also expressed by the PP cells, and of Sox 4 at the anterior intestinal portal and NFAT, critical factors in valvulogenesis. CONCLUSIONS: Cells existing already during gastrulation are associated with valve formation days later. The Wnt/beta-catenin signaling in PP cells is normally repressed by Wnt antagonists and Hex is up-regulated. The antagonism occurring at the receptor level is bypassed by Li exposure by its intracellular inactivation of GSK-3 directly to augment Wnt signaling. PMID- 18418888 TI - Autologous synovial fluid enhances migration of mesenchymal stem cells from synovium of osteoarthritis patients in tissue culture system. AB - Synovial fluid from osteoarthritic knee contains mesenchymal stem cells (MSCs). One of the possible reservoirs of MSCs in synovial fluid is synovial tissue, and synovial fluid may induce mobilization of MSCs into synovial fluid in osteoarthritis patients. Here, we investigated whether synovial fluid expanded synovial MSCs in a tissue culture system. Human synovium and synovial fluid were obtained from osteoarthritis patients during total knee arthroplasties. In the tissue culture system, autologous synovial fluid expanded synovial cells statistically higher than alpha MEM + FBS, and the addition of TGF beta 3 to alpha MEM + FBS increased expansion to a similar level in all 11 donors. The addition of decorin or anti-TGFbeta neutralizing antibody to synovial fluid partially inhibited synovial cell expansion. In cell culture assay, synovial fluid proliferated synovial cells fewer than alpha MEM + FBS. The expanded synovial cells in synovial fluid retained multipotentiality and showed surface markers similar to those of MSCs. We demonstrated that autologous synovial fluid enhanced expansion of MSCs in tissue culture of synovium from osteoarthritis patients by promoting cell migration. This effect was partially affected by TGFbeta. PMID- 18418889 TI - Subcellular distribution of the insulin-like growth factor (IGF) binding proteins (IGFBPs) 2 and 3 in articular chondrocytes. AB - The insulin-like growth factor (IGF) is a major anabolic regulator in articular cartilage. The IGF-binding proteins (IGFBPs) are increased during osteoarthritis (OA), but the function of the later proteins remains unknown. In general, the IGFBPs are pluripotential effectors capable of IGF regulation and of acting on their own to control key cell functions, including survival and proliferation. The independent functions are often associated with their cell location, and therefore this study explores the distribution of IGFBP-2 and IGFBP-3 in articular chondrocytes. Immunohistochemistry was used to localize IGFBP-2 in normal human articular cartilage. Bovine chondrocytes were used for subcellular fractionation (hypotonic cell lysis) under nonreducing conditions and nuclear purification (centrifugation on sucrose cushions). Cell fraction markers and IGFBPs were assayed in the subcellular fractions by Western immunoblot. The IHC results showed association of IGFBP-2 with chondrocytes, but not with the nuclei. Subcellular fractionation of isolated chondrocytes yielded intact nuclei as assessed at the light microscopic level; the nuclear marker histone H1 was exclusively associated with this fraction. More than 90% of the cytoplasmic marker GAPDH and all the detectable IGFBP-2 were in the cytoplasmic fraction. Immunoreactive IGFBP-3 was found in the cytoplasmic and peri-nuclear/nuclear fractions. Chondrocytes contain intracellular IGFBP-2 and IGFBP-3 but only IGFBP 3 is associated with nuclei. This suggests the hypothesis that the actions of these IGFBPs in articular cartilage extend beyond the classic modulation of IGF receptor action. PMID- 18418890 TI - A focus on high-content cytometry. PMID- 18418891 TI - Induction of drug metabolism enzymes and transporters by oltipraz in rats. AB - Coordinate regulation of Phase-I and -II enzymes with xenobiotic transporters has been shown after treatment with microsomal enzyme inducers. The chemopreventive agent oltipraz (OPZ) induces Phase-I and -II drug-metabolizing enzymes such as CYP2B and NQO1. The purpose of this study was to examine the regulation of drug metabolizing enzymes and transporters in response to OPZ treatment and to investigate a potential role for constitutive androstane receptor (CAR) in OPZ mediated induction. Sprague-Dawley rats treated with OPZ exhibited increased mRNA and protein levels of both Nqo1 and Cyp2b1/2 by 24 h. To examine whether OPZ activates transporter gene expression via CAR, sexually dimorphic male and female Wistar-Kyoto (WKY) rats were treated with OPZ and mRNA levels quantified by bDNA signal amplification. OPZ induced Ugt1a6 and Ugt2b1 in males significantly higher than in females, indicating a CAR-dependent mechanism of induction. However, OPZ induced microsomal epoxide hydrolase, NAD(P)H quinone oxidoreductase, and Cyp3a1/23 equally in both genders, indicating a CAR-independent mechanism of induction of these genes. Similarly, the transporters Mdr1a, Mdr1b, Mrp3, and Mrp4 were induced by OPZ without any apparent difference between genders. In summary, OPZ coordinately increases multiple hepatic xenobiotic transporter mRNA levels, along with Phase-I and -II enzymes some of which may occur through CAR dependent mechanisms. PMID- 18418892 TI - Differential effect of ascorbic acid and n-acetyl-L-cysteine on arsenic trioxide mediated oxidative stress in human leukemia (HL-60) cells. AB - Arsenic trioxide (ATO) has been recommended for the treatment of refractory cases of acute promyelocytic leukemia (APL). Recent studies in our laboratory indicated that oxidative stress plays a key role in ATO-induced cytotoxicity in human leukemia (HL-60) cells. In the present investigation, we performed the MTT assay and trypan blue exclusion test for cell viability. We also performed the thiobarbituric acid test to determine the levels of malondialdehyde (MDA) production in HL-60 cells coexposed to either ascorbic acid (AA) and ATO or to n acetyl-L-cysteine (NAC) and ATO. The results of MTT assay indicated that AA exposure potentiates the cytotoxicity of ATO in HL-60 cells, as evidenced by a gradual increase in MDA levels with increasing doses of AA. In contrary, the addition of NAC to ATO-treated HL-60 cells resulted in a dose-dependent decrease of MDA production. From these results, we conclude that the addition of the AA to ATO-treated HL-60 cells enhances the formation of reactive oxygen species (ROS), whereas the addition of NAC under the same experimental condition significantly (p < .05) decreases the level of ROS formation. On the basis of these direct in vitro findings, our studies provide evidence that AA may extend the therapeutic spectrum of ATO. The coadministration of NAC with ATO shows a potential specificity for tumor cells, indicating that it may not enhance the clinical outcome associated with ATO monotherapy in vivo. PMID- 18418893 TI - Aluminum-mediated metabolic changes in rat serum and urine: a proton nuclear magnetic resonance study. AB - The toxic effects of Al(3+) have been studied in 90-days AlCl(3) orally treated male albino rats (n = 7) using (1)H NMR spectroscopy-based metabolic profile of rat serum and urine, serum enzyme tests, behavioral impairment, and histopathology of kidney and liver. Metabolic profile of 90-days Al(3+)-treated rat sera showed significantly elevated levels of alanine, glutamine, beta-hydroxy butyrate, and acetoacetate and significantly decreased level of acetone when compared with that of control rats. However, metabolic profile of 90-days Al(3+) treated rat urine showed significantly decreased levels of citrate, creatinine, allantoin, trans-aconitate, and succinate and significantly increased level of acetate when compared to control rats. The overall perturbations observed in the metabolic profile of serum and urine demonstrate the impairment in the tricarboxylic acid cycle, liver and kidney metabolism, which was further reinstated by clinical chemistry and histopathological observations. Moreover, "in vivo" behavioral impairment has also been observed as the indication of aluminum neurotoxicity. PMID- 18418896 TI - Modulatory potential of ellagic acid, a natural plant polyphenol on altered lipid profile and lipid peroxidation status during alcohol-induced toxicity: a pathohistological study. AB - Polyphenol-rich dietary foodstuffs, consumed as an integral part of vegetables, fruits, and beverages have attracted attention due to their antioxidant and anticancer properties. Ellagic acid (EA), a polyphenolic compound widely distributed in fruits and nuts, has been reported to scavenge free radicals and inhibit lipid peroxidation. Chronic consumption of alcohol potentially results in serious illness including hepatitis, fatty liver, hypertriglyceridemia, and cirrhosis. A little is known about the influence of EA on alcohol toxicity in vivo. Accordingly, in the present study, we have evaluated the protective effects of EA on lipid peroxidation and lipid levels during alcohol-induced toxicity in experimental rats. Forty female albino Wistar rats, which were weighing between 150-170 g were used for the study. The toxicity was induced by administration of 20% alcohol orally (7.9 g/kg body wt.) for 45 days. Rats were treated with EA at three different doses (30, 60, and 90 mg/kg body wt.) via intragastric intubations together with alcohol. At the end of experimental duration, liver marker enzymes (i.e., aspartate transaminase, alanine transaminase), lipid peroxidative indices (i.e., thiobarbituriacid reactive substances and hydroperoxides) in plasma, and lipid levels (i.e., cholesterol, free fatty acids, triglycerides and phospholipids) in tissues were analyzed to evaluate the antiperoxidative and antilipidemic effects of EA. Liver marker enzymes, lipid peroxidative indices, and lipid levels, i.e., cholesterol, triglycerides and free fatty acids, were significantly increased whereas phospholipid levels were significantly decreased in the alcohol-administered group. EA treatment resulted in positive modulation of marker enzymes, peroxidative indices, and lipid levels. EA at the dose of 60 mg/kg body wt. was found to be more effective when compared to the other two doses. Histological changes observed were also inconsistent with the biochemical parameters. Our study suggests that EA exerts beneficial effects at the dosage of 60 mg/kg body wt. against alcohol-induced damage, and it can be used as a potential drug for the treatment of alcohol-abuse ailments in the near future. PMID- 18418895 TI - Gender divergent expression of Nqo1 in Sprague Dawley and August Copenhagen x Irish rats. AB - In the mammalian liver, there is an abundance of enzymes that function to enable the safe and efficient elimination of potentially harmful xenobiotics that are encountered through environmental exposure. A variety of factors, including gender and genetic polymorphisms, contribute to the variation between an individual system's detoxification capacity and thus its ability to protect itself against oxidative stress, cellular damage, cell death, etc. NAD(P)H:quinone oxidoreducatase 1 (Nqo1) is an antioxidant enzyme that plays a major role in reducing reactive electrophiles, thereby protecting cells from free radical damage and oxidative stress. The goal of this study was to determine the gender-specific expression and inducibility of Nqo1 in the Sprague Dawley (SD) and August Copenhagen x Irish (ACI) rat strains, two strains that are commonly used in drug metabolism and drug-induced enzyme induction, toxicity, and carcinogenesis studies. Nqo1 mRNA, protein, and activity levels were determined through 96 h in SD and ACI males and females following treatment with known Nqo1 inducers oltipraz and butylated hydroxyanisole. In the SD strain, gender dimorphic expression of Nqo1 was observed with female mRNA, protein, and activity levels being significantly higher than in males. In contrast, there were minimal differences in Nqo1 mRNA, protein, and activity levels between ACI males and females. The gender dimorphic expression of Nqo1 in the SD rats was maintained through the course of induction, with female-induced levels greater than male induced levels indicating that SD females may have a greater capacity to protect against oxidative stress and thus a decreased susceptibility to carcinogens. PMID- 18418897 TI - Local circuitry in the anterior caudal lobe of the mormyrid cerebellum: a study of intracellular recording and labeling. AB - The caudal lobe of the mormyrid cerebellum includes the anterior portion, which is associated with the lateral line and eighth nerve senses, and the posterior portion, which is associated with the electrosense. This study examines the physiology and morphology of cells in the anterior portion in slice preparations. Two subtypes of Purkinje cells, efferent cells and stellate cells, are described. Multipolar Purkinje cells are located in the central region of the lobe, with large, multipolar, spiny dendrites and locally ending axons. Small Purkinje cells are located along its anterior border with the eminentia granularis anterior (EGa), with spiny dendrites in the molecular region. Axons of some small Purkinje cells end locally, whereas axons of other such cells are cut at the surface of the slices, suggesting that they project outside the lobe. Efferent cells are also distributed along the border with EGa. These cells have thin, smooth dendrites in the molecular region, and their axons are cut at the sliced surface. Stellate cells have thin, smooth dendrites and locally terminating axons. Physiologically, all types of cells respond to parallel fiber activation, but only multipolar Purkinje cells showed characteristic all-or-none climbing fiber responses. Although the majority of Purkinje cells fire a single type of spikes at resting level, a subset of small Purkinje cells fire small, narrow and large, broad spikes. Thus, the anterior caudal lobe of the mormyrid cerebellum is different from the mammalian cerebellum in having different subtypes of Purkinje cells and local termination of many Purkinje cell axons. PMID- 18418898 TI - Ion transport peptide splice forms in central and peripheral neurons throughout postembryogenesis of Drosophila melanogaster. AB - Ion transport peptides (ITPs) belong to a large arthropod neuropeptide family including crustacean hyperglycaemic hormones and are antidiuretic hormones in locusts. Because long and short ITP isoforms are generated by alternative splicing from a single gene in locusts and moths, we investigated whether similarly spliced gene products occur in the nervous system of Drosophila melanogaster throughout postembryogenesis. The itp gene CG13586 was reanalyzed, and we found three instead of the two previously annotated alternatively spliced mRNAs. These give rise to three different neuropeptides, two long C-terminally carboxylated isoforms (DrmITPL1 and DrmITPL2, both 87 amino acids) and one short amidated DrmITP (73 amino acids), which were partially identified biochemically. Immunocytochemistry and in situ hybridization reveal nine larval and 14 adult identified neurons: four pars lateralis neurosecretory neurons, three hindgut innervating neurons in abdominal ganglia, and a stage-specific number of interneurons and peripheral bipolar neurons. The neurosecretory neurons persist throughout postembryogenesis, form release sites in corpora cardiaca, and invade corpora allata. One type of ITP-expressing interneuron exists only in the larval and prepupal subesophageal ganglia, whereas three types of interneurons in the adult brain arise in late pupae and invade circumscribed neuropils in superior median and lateral brain areas. One peripheral bipolar and putative sensory neuron type occurs in the larval, pupal, and adult preterminal abdominal segments. Although the neurosecretory neurons may release DrmITP and DrmITPL2 into the haemolymph, possible physiological roles of the hindgut-innervating and peripheral neurons as well as the interneurons are yet to be identified. PMID- 18418899 TI - Selective activation of NF-kappaB and E2F by low concentration of arsenite in U937 human monocytic leukemia cells. AB - Arsenite has been reported to exert dose-dependent dual effects: triggering apoptosis at relatively high concentrations, whereas inducing partial differentiation at low concentrations in leukemia cells. However, the relevant molecular mechanisms of its action at low and nonapoptotic concentrations remain to be elucidated. We examined the effect of arsenite on activation of key transcription factors in cultured U937 human monocytes/macrophages. Electrophoretic mobility shift assay (EMSA), protein/DNA array and luciferase reporter assay were used to analyze the effect of arsenite on the functional activities of transcription factors. Protein/DNA array analysis showed that activation of E2F was seen after 6-h exposure to 1 and 10 microM arsenite. In contrast, activation of NF-kappaB took place only at 1 microM arsenite, whereas 10 microM arsenite showed no recognizable effect on this nuclear transcription factor in the protein/DNA array analysis. EMSA using a NF-kappaB consensus probe indicates the functional activation of RelB/p50 in the presence of 1 microM arsenite, confirming the above results. Luciferase reporter assay for NF-kappaB showed activation of NF-kappaB in the presence of 1 microM arsenite. Interleukin (IL)-8 and B-cell-activating factor of the tumor necrosis factor family (BAFF) mRNA expression, which have been shown to be regulated through NF-kappaB, were activated in the presence of 1 microM arsenite. These results support the hypothesis that the primary action of nonapoptotic concentrations of arsenite in this cell line is activation of NF-kappaB, signaling as a decision maker for end results such as inflammation disease or cancer. This finding offers the possibility of providing a logical explanation for the observations made by many scientists that chronic exposure of human populations to low doses of arsenic is significantly correlated to clinical signs of inflammation in many tissues. PMID- 18418900 TI - Distribution and neurochemical identification of pancreatic afferents in the mouse. AB - Dysfunction of primary afferents innervating the pancreas has been shown to contribute to the development of painful symptoms during acute and chronic pancreatitis. To investigate the distribution and neurochemical phenotype of pancreatic afferents, Alexa Fluor-conjugated cholera toxin B (CTB) was injected into the pancreatic head (CTB-488) and tail (CTB-555) of adult male mice to label neurons retrogradely in both the dorsal root ganglia (DRG) and nodose ganglia (NG). The NG and DRG (T5-T13) were processed for fluorescent immunohistochemistry and visualized by using confocal microscopy. Spinal pancreatic afferents were observed from T5 to T13, with the greatest contribution coming from T9-T12. The pancreatic afferents were equally distributed between right and left spinal ganglia; however, the innervation from the left NG was significantly greater than from the right. For both spinal and vagal afferents there was significantly greater innervation of the pancreatic head relative to the tail. The total number of retrogradely labeled afferents in the nodose was very similar to the total number of DRG afferents. The neurochemical phenotype of DRG neurons was dominated by transient receptor potential vanilloid 1 (TRPV1)-positive neurons (75%), GDNF family receptor alpha-3 (GFRalpha3)-positive neurons (67%), and calcitonin gene related peptide (CGRP)-positive neurons(65%) neurons. In the NG, TRPV1-, GFRalpha3-, and CGRP-positive neurons constituted only 35%, 1%, and 15% of labeled afferents, respectively. The disparity in peptide and receptor expression between pancreatic afferents in the NG and DRG suggests that even though they contribute a similar number of primary afferents to the pancreas, these two populations may differ in regard to their nociceptive properties and growth factor dependency. PMID- 18418901 TI - Sphingosine can pre- and post-condition heart and utilizes a different mechanism from sphingosine 1-phosphate. AB - Consistent with previous reports, sphingosine at a high concentration (5 microM) was cardiotoxic as evidenced by increased infarct size in response to ischemia/reperfusion in an ex vivo rat heart. Sphingosine 1-phosphate (S1P) at 5 microM was cardioprotective. However, at a physiologic concentration (0.4 microM) sphingosine as well as S1P was effective in protecting the heart from ischemia/reperfusion injury both when perfused prior to 40 min of ischemia (preconditioning) or when added to reperfusion media following ischemia (postconditioning). Protection by sphingosine and S1P was evidenced with both pre and post-conditioning by a >75% recovery of left ventricular developed pressure during reperfusion and a decrease in infarct size from 45% of the risk area to less than 8%. When VPC23019, an S1P(1and3)G-protein coupled receptor antagonist, was added to the preconditioning or postconditioning medium along with S1P, it completely blocked S1P-induced protection. However, VPC 23019 did not affect the ability of 0.4 microM sphingosine to either precondition or postcondition hearts. Studies of preconditioning revealed that inhibition of protein kinase C with GF109203X blocked preconditioning by S1P. However, GF109203X did not affect preconditioning by 0.4 microM sphingosine. Likewise, cotreatment with the PI3 kinase inhibitor wortmanin blocked preconditioning by S1P but not by sphingosine. By contrast, inhibition of protein kinase G with KT5823 had no effect on S1P preconditioning but completely eliminated preconditioning by sphingosine. Also, the protein kinase A inhibitory peptide 14-22 amide blocked preconditioning by sphingosine but not S1P. These data reveal for the first time that sphingosine is not toxic at physiologic concentrations but rather is a potent cardioprotectant that utilizes a completely different mechanism than S1P; one that is independent of G-protein coupled receptors and utilizes cyclic nucleotide-dependent pathways. PMID- 18418903 TI - Eosinophilia with stroke in a Chinese patient: Kimura's disease complicated with fibroblastic endocarditis, first report. PMID- 18418904 TI - How I became a biochemist/molecular biologist. PMID- 18418905 TI - FDA notifications. Tentative approval for generic nevirapine tablets. PMID- 18418902 TI - Age-related changes to tumor necrosis factor receptors affect neuron survival in the presence of beta-amyloid. AB - Inflammation including local accumulations of tumor necrosis factor alpha (TNF alpha) is a part of Alzheimer's disease pathology and may exacerbate age-related neurodegeneration. Most studies on TNF-alpha and TNF neuronal receptors are conducted by using embryonic neurons. Few studies consider age-related deficits that may occur in neurons. Age-related changes in susceptibility to TNF-alpha through TNF receptor 1 (TNFR1) and receptor 2 (TNFR2) expression could increase susceptibility to beta-amyloid (1-42, Abeta42). Evidence is conflicting about which receptor mediates survival and/or apoptosis. We determined how aging affects receptor expression in cultured adult rat cortical neurons. Old neurons were more susceptible to Abeta42 toxicity than middle-aged neurons, and the addition of TNF-alpha was neuroprotective in middle-aged neurons, but exacerbated the toxicity from Abeta42 in old neurons. These pathologic and protective responses in old and middle-aged neurons, respectively, correlated with higher starting TNFR1 and TNFR2 mRNA levels in old vs. middle-aged neurons. Middle-aged neurons treated with TNF-alpha plus Abeta42 did not show an increase in either TNFR1 or TNFR2 mRNA, but old neurons showed an up-regulation in TNFR2 mRNA and not TNFR1 mRNA. Despite these mRNA changes, surface immunoreactivity of both TNFR1 and TNFR2 increased with the dose of TNF-alpha in middle-aged neurons. However, middle-aged neurons treated with TNF-alpha plus Abeta42 showed an up regulation in both TNFR1 and TNFR2 surface expression, whereas old neurons failed to up-regulate surface expression of either receptor. These findings support the hypothesis that age-related changes in TNF-alpha surface receptor expression contribute to the neuronal loss associated with inflammation in Alzheimer's disease. PMID- 18418906 TI - Breast cancer risk factors. The latest facts, your best means of defense. PMID- 18418907 TI - FDA approves new drug for specific type of advanced breast cancer. PMID- 18418908 TI - Hormone patches, gels may be less likely than pills to cause blood clots. PMID- 18418909 TI - Memorial issue in honor of Dr. Setsuro Ebashi. PMID- 18418910 TI - [A 47-old woman with acute confusion following total hip arthroplasty]. PMID- 18418911 TI - [In-hospital stroke]. PMID- 18418912 TI - [HIV infections more frequent among black homosexuals]. PMID- 18418913 TI - Bibliography. Current world literature. Genitourinary systems. PMID- 18418914 TI - Spitz nevus with rosette-like structures: a new histologic variant. PMID- 18418915 TI - [Should volatile anaesthetics decrease morbidity and mortality after cardiac surgery? Comments of three recent meta-analysis]. PMID- 18418916 TI - The end of the journal, as we know it: commentary. PMID- 18418917 TI - NJ: Post-seizure pt. falls on way to bathroom: court upheld verdict for defendant hospital. PMID- 18418918 TI - [Extrasecretory liver function disorder in patients with chronic pancreatitis]. AB - Bile production and bile secretion studies in 112 patients with primary chronic pancreatitis have demonstrated. Duodenal intubational chromatic examination can be used in addition to standard laboratory and device methods for early diagnosis of "biliary insuffiency" and cholelithiasis. The analysis of efficacy of Ursofalk was made in 30 patients with chronic pancreatitis. Patients received Ursofalk in a dose 10 mg/kg/day for one month. It was established that Ursofalk stabilizes bile secretion and removes biliary insuffiency. PMID- 18418919 TI - [The clinico-functional characteristics of the pancreas in chronic obstructive pulmonary disease]. AB - The authors conducted a complex examination of functional condition of pancreas in 70 patients with chronic obstructive pulmonary disease (COPD). The defeat of pancreas was revealed in 44.3% patients with COPD. The rate and degree of pancreatic defeat depended of severity and long of basic disease. The pancreatic defeat often had subclinical character and was revealed after complex examination of patient. It was shown that secondary erythrocytosis, changes of microcyrculation and haemostas system to hypercoagulation have important role in disturbance of pancreatic function. PMID- 18418920 TI - [Smoking as a possible factor of chronic pancreatitis and pancreatic cancer (literature review)]. PMID- 18418921 TI - [The duodenal wall injury as a cause of acute pancreatitis]. AB - Dynamics of cholecystokinin-like, secretin and somatostatin immunoreactivity of blood plasma and extracts of duodenal mucosa, frequency of acute pancreatitis (AP) after the resection of the duodenum was studied on not purebred dogs. Rising of a secretin concentration in blood plasma after the clinoid resection of the duodenum, accompanied by decreasing of its concentration in duodenal mucosa, was revealed. Decreasing of cholecystokinin concentration in a neutral extract was revealed. In 24 hours alter resection of the duodenum AP was observed in 38% of cases, infusion of the lorglumide was reduced the frequency of this complication up to 6.2% (p<0.05). PMID- 18418922 TI - [Dynamics of indices of visceral sensitivity in patients with irritable bowel syndrome treated with spasmolytics]. PMID- 18418923 TI - [Deviation in protein content of blood serum in patients with non-Hodgkin's lymphoma]. AB - The author observed 20 patients with non-Hodgkin's lymphoma of high, medium and low degree of malignancy for period of time from 1996 till 1998. Deviation in albumin and globulin fractions as well as such tumor-associated antigens as alpha fetoprotein and cancer-embryonal were observed. Analysis and assessment of clinical and laboratory results show that degree of albumin and globulin deviation in patients with non-Hodgkin's lymphoma has the tendency for change depending on a stage of the disease, degree of malignancy, sex and age of patients. PMID- 18418925 TI - [Improvement of the quality of life in patients with non-operable pancreatic cancer treated with polyplatillen]. AB - The authors summarize in the article experience of a new anticancer drug Polyplatillen use in the chemotherapy of advanced pancreatic cancer. Polyplatillen use in the palliative chemotherapy in the patients with advanced pancreatic cancer permits to realize a control of pain and stabilize an ECOG status. Tendency to the improvement of total survival during an early period was registered in the group of patients with advanced pancreatic cancer who were treated with Polyplatillen. PMID- 18418924 TI - [Effect of riodipin on the central and cerebral haemodynamics, cell membrane permeability in patients with hypertensive disease]. AB - The author studied features of indices of the central, cerebral haemodynamics and penetrability of cellular membranes in patients with hypertensive disease during the treatment of the disease through oral and transdermal formulation of riodipin. It was established that efficiency of the treatment increases when transdermal formulation of riodipin was used. PMID- 18418926 TI - [Application of nootropic agents in complex treatment of patients with concussion of the brain]. AB - 65 patients with a mild craniocereberal trauma have been observed. Medical examination included among general clinical methods the following methods: KT (MRT) of the brain, oculist examination including the observation of eye fundus. For objectification of a patient' complaints the authors used orientation and Galvestona's amnesia tests, feeling scale (psychological test), the table to determine the level of memory. Tests have been carried out on the first, tenth and thirty day of the treatment. Patients of the first group received in a complex treatment -pramistar, patients of the second group - piracetam. Patients of both groups noted considerable improvement during a complex treatment (disappearance of headache, dizziness and nausea) and at the same time patients receiving pramistar had better restoration of orientation and feeling. Pramistar was also more effective in patients with amnesia. PMID- 18418927 TI - [Use of the electrophoresis with acetic acid in a complex treatment of heel spurs at a sanatorium]. AB - The author studied the influence of physical factors of electrophoresis with acetic acid, massage and laseropuncture in sanatorium conditions where 28 patients with heeling spurs have been treated. The patients in a control group (n=25) received a traditional treatment - an injection of hydrocortisone in area of heeling spurs, skin rubbing with herbs (tincture with different flowers, elder berry buds. Efficiency of a proposed medical complex was shown using clinical and radiographic data. PMID- 18418928 TI - [Molecule of taurine and its quantum-pharmacological properties]. AB - Quantum-pharmacological properties of taurine and its zwitterion form have been studied. Geometrical and energetic parameters were calculated using ab initio 6 31G* technique. These parameters are the following: distance between atoms, distribution of electronic density only of external valency electrons, distribution of electrostatic potential, atomic charges, dipole moment, localization and energy of high and low occupied and low unoccupied molecular orbitals. Authors established that the sulfonate group is a electrophilic center of taurin molecul and amide group is a nucleophilic center. The biggest value of electrostatic potential is located around sulfonate group. The big value of energy of low occupied molecular orbital of taurin gives grounds to consider this substance as a reducing reagent and explains its antioxidant properties. PMID- 18418929 TI - Provision of products: should we be concerned? PMID- 18418930 TI - A service evaluation of the use of silicone-based adhesive remover. AB - Evidence-based practice has become a priority for nurses, but until the publication of Caring for Stoma Patients, Best Practice Guidelines (Clinimed Resource for Education and Specialist Training, 2006), there was very little evidence published for many aspects of stoma care nursing. This document provides best practice information for many specific areas of stoma care, but there is still a lack of evidence on which clinical decisions regarding subjective issues can be based. As a result, the stoma care department at St George's Healthcare NHS Trust undertook a study on the use of a silicone-based adhesive remover. Dykes (2001) highlights the trauma that can occur as a result of skin stripping during pouch removal. The use of a silicone-based adhesive remover allows for the rapid and painless removal of a stoma pouch without the associated problems of skin stripping (Cutting, 2006). Results from the study conclude that patients experience great benefit from the use of a silicone-based adhesive remover and that all patients should be offered the use of one. PMID- 18418931 TI - Flatus, odour and the ostomist: coping strategies and interventions. AB - Both new and established ostomists fear the loss of control over their bodily function that stoma surgery brings about. They often express feelings of embarrassment and isolation as they try and adjust to their new lifestyle. Quality-of-life studies often highlight flatus and odour to be two of the top five fears or worries among ostomists. This article reviews the current literature offering an insight into how flatus and odour impacts on an ostomist's life and considers interventions to assist the nurse in aiding the ostomist through this period of adaptation. PMID- 18418932 TI - Parastomal hernia: incidence, prevention and treatment strategies. AB - Parastomal hernia continues to be a common and distressing problem for patients with stomas, and research investigating prevention strategies is scant. In March 2005 Thompson and Trainor reported that the introduction of a prevention programme for 1-year post-stoma surgery formation had significantly reduced the incidence of development of parastomal hernia. This was further supported by a follow-up study in 2007, strengthening the reliability and validity of the first findings by confirming a statistically significant reduction in the incidence of parastomal hernias through the introduction of a simple non-invasive prevention programme. This article reviews the current literature on incidence, prevention and treatment, together with a step-by-step guide for stoma care nurses to implement the prevention programme and/or study within their area. PMID- 18418933 TI - Providing dietary advice for the individual with a stoma. AB - Concerns over diet and nutrition are common among ostomists and their carers. It is important for healthcare professionals involved in the care of patients with a colostomy, ileostomy or urostomy to appreciate that stoma surgery changes the body's usual processes of nutritional absorption and excretion, so that informed support and advice on diet can be provided. A considerable proportion of patients admitted to hospital in the United Kingdom are malnourished, or at risk of becoming so. Patients undergoing gastrointestinal surgery resulting in stoma formation are at particular risk as a result of the potential for preoperative malnutrition caused by the effects of their underlying disease and prolonged periods of fasting during the immediate pre- and postoperative period. Screening and monitoring of the nutritional status of individuals with a stoma should be an ongoing process, beginning preoperatively and continuing after discharge from hospital. This article will examine the effects of stoma formation on diet and consider ostomists' potential for altered absorption of nutrients. Current issues relating to patient nutrition in the hospital setting, their relevance for stoma patients and the role of the nurse in the provision of nutritional care will also be discussed. PMID- 18418936 TI - [Rabbit IgG antibodies against phospholipase A2 from Crotalus durisms terrificus neutralize the lethal activity of the venom]. PMID- 18418938 TI - Global warming in Wisconsin. PMID- 18418937 TI - Studies unable to prove prayer is helpful for patients. PMID- 18418941 TI - Exercising safely with CVD. PMID- 18418943 TI - Breast cancer screening gets better. PMID- 18418942 TI - Breathing easier with COPD. PMID- 18418944 TI - Fibromyalgia: understanding a mysterious ailment. PMID- 18418946 TI - Retraction: Wang, Yao et al. The flhDC gene affects motility and biofilm formation in Yersinia pseudotuberculosis. Sci China Ser C-Life Sci, 2007, 50(6): 814-821. PMID- 18418945 TI - I have Dupuytren's disease. Is there anything I can do, short of surgery, to treat or cure this? PMID- 18418947 TI - Lentiviral vector-mediated reporter gene expression in avulsed spinal ventral root is short-term, but is prolonged using an immune "stealth" transgene. AB - PURPOSE: Spinal root avulsions result in paralysis of the upper and/or lower extremities. Implanting a peripheral nerve bridge or reinsertion of the avulsed roots in the spinal cord are surgical strategies that lead to some degree of functional recovery. In the current study lentiviral (LV) vector-mediated gene transfer of a green fluorescent protein (GFP) reporter gene was used to study the feasibility of gene therapy in the reimplanted root to further promote regeneration of motor axons. METHODS: A total of 68 female Wistar rats underwent unilateral root avulsion of the L4, L5 and L6 ventral lumbar roots. From 23 rats intercostal nerves were dissected before ventral root avulsion surgery, injected with a lentiviral vector encoding GFP (LV-GFP) and inserted between the spinal cord and avulsed rootlet. In the remaining 45 rats, the avulsed ventral root was injected with either LV-GFP or a lentiviral vector encoding a fusion between a GlyAla repeat and GFP (LV-GArGFP), and reinserted into the spinal cord. Expression of GFP was evaluated at 1,2, 4 and 10 weeks, and one group at 4 months. RESULTS: LV-GFP transduction of either nerve implants or reimplanted ventral roots revealed high GFP expression during the first 2 post-lesion weeks, but virtually no expression at 4 weeks. Since this reduction coincided with the appearance of mononuclear cells at the repair site, an immune response against GFP may have occurred. In a subsequent experiment reimplanted ventral roots were transduced with a vector encoding GFP fused with the GlyAla repeat of Epstein Barr virus Nuclear Antigen 1 known to prevent generation of antigenic peptides from transgene products. Expression of this "stealth" gene persisted for at least 4 months in the reimplanted root. CONCLUSION: Thus persistent transgene expression can be achieved with non-immunogenic transgene products in reimplanted ventral roots. This demonstrates the feasibility of combining neurosurgical repair with LV vector-mediated gene therapy. The current approach will be used in future experiments with LV vectors encoding neurotrophic factors to enhance the regeneration of spinal motor neurons after traumatic avulsion of spinal nerve roots. PMID- 18418948 TI - Concerning the article by Lotze et al., 2006: combination of TMS and fMRI reveals a specific pattern of reorganization in M1 in patients after complete spinal cord injury. PMID- 18418949 TI - Capillary electrophoresis. Methods and protocols. Preface. PMID- 18418950 TI - New radon data for England and Wales. PMID- 18418951 TI - Advice on risks from tritium. PMID- 18418952 TI - The environmental impact of growth-promoting compounds employed by the United States beef cattle industry: history, current knowledge, and future directions. AB - The current state of knowledge regarding the environmental impact of growth promoting compounds associated with the U.S. beef cattle industry is extensive in some areas but virtually nonexistent in others. The compounds administered to the cattle are quite well understood, as are bovine metabolism and excretion. If the sex and age of the cattle on the feedlot are known, the metabolites excreted by the cattle should be predictable with a great deal of accuracy. The fate, transport, and biological effects of growth-promoting compounds are just beginning to be studied. Most of the research conducted on the fate and transport of growth-promoting compounds has focused on 17beta-E2; however, much of this research was not conducted using feedlot runoff or manure. Studies are needed that focus specifically on manures and runoff from experimental or commercial feedlots. To date, the degree to which growth-promoting compounds are released from feedlots in a bioavailable form remains a point of speculation. The environmental fate and transport of TBA, P, and MGA have not been well studied. Comparisons between the fate and transport of T and 17beta-E2, however, make it clear that compounds with similar structure may behave very differently once released into the environment. Considering that 17beta-E2 is a naturally occurring estrogen and that TBA is a nonaromatizable androgen, it is not surprising that these compounds directly impact the reproductive physiology of fishes. The effects of these two compounds have been well documented, as has been described here; however, the effects of P and MGA exposures have gone largely uninvestigated. This is a serious critical gap in our knowledge base because progestogins play an important role in sex steroid synthesis and reproduction. Clearly, additional research on the consequences of exposures to P and MGA is warranted. The majority of research investigating the effects of 17beta-E2 and TBA metabolites on fish has been conducted in the laboratory and has typically focused on continuous, pharmacological exposures to single compounds. These exposures may not bear much similarity to environmentally relevant exposures, and as such may offer little information regarding biological effects seen in nature. Cattle feedlot runoff is likely to contain a suite of growth-promoting compounds rather than any single compound. Clearly, deciphering the biological effects of exposure to complex mixtures containing androgenic, estrogenic, and progestogenic compounds will remain an important area of study for the next few years. A second complexity associated with the biological runoff from cattle feedlots is the discontinuous nature of the release. It is likely that inadvertent entry of growth-promoting compounds will follow spring snowmelt or rainstorm events. These events will result in intermittent, pulsed exposures to high concentrations of these compounds interspersed by long-term exposures to lower concentrations. The effects of exposure timing and duration should be considered to generate a clearer understanding of the biological consequences of exposures to growth promoting compounds. To date, a very limited number of studies (only one!) have sought to determine whether fish living in waterways receiving runoff from cattle feedlots are adversely affected by growth-promoting compounds associated with the runoff. Clearly, more field studies need to be conducted before a relationship between cattle feedlot effluent and biological consequences can be elucidated. PMID- 18418953 TI - Arsenic in marine mammals, seabirds, and sea turtles. AB - Although there have been numerous studies on arsenic in low-trophic-level marine organisms, few studies exist on arsenic in marine mammals, seabirds, and sea turtles. Studies on arsenic species and their concentrations in these animals are needed to evaluate their possible health effects and to deepen our understanding of how arsenic behaves and cycles in marine ecosystems. Most arsenic in the livers of marine mammals, seabirds, and sea turtles is AB, but this form is absent or occurs at surprisingly low levels in the dugong. Although arsenic levels were low in marine mammals, some seabirds, and some sea turtles, the black footed albatross and hawksbill and loggerhead turtles showed high concentrations, comparable to those in marine organisms at low trophic levels. Hence, these animals may have a specific mechanism for accumulating arsenic. Osmoregulation in these animals may play a role in the high accumulation of AB. Highly toxic inorganic arsenic is found in some seabirds and sea turtles, and some evidence suggests it may act as an endocrine disruptor, requiring new and more detailed studies for confirmation. Furthermore, DMA(V) and arsenosugars, which are commonly found in marine animals and marine algae, respectively, might pose risks to highly exposed animals because of their tendency to form reactive oxygen species. In marine mammals, arsenic is thought to be mainly stored in blubber as lipid-soluble arsenicals. Because marine mammals occupy the top levels of their food chain, work to characterize the lipid-soluble arsenicals and how they cycle in marine ecosystems is needed. These lipid-soluble arsenicals have DMA precursors, the exact structures of which remain to be determined. Because many more arsenicals are assumed to be present in the marine environment, further advances in analytical capabilities can and will provide useful future information on the transformation and cycling of arsenic in the marine environment. PMID- 18418954 TI - Environmental chemistry, ecotoxicity, and fate of lambda-cyhalothrin. AB - Lambda-cyhalothrin is a pyrethroid insecticide used for controlling pest insects in agriculture, public health, and in construction and households. Lambda cyhalothrin is characterized by low vapor pressure and a low Henry's law constant but by a high octanol-water partition coefficient (K(ow)) and high water-solid organic carbon partition coefficient (K(oc)) values. Lambda-cyhalothrin is quite stable in water at pH < 8, whereas it hydrolyzes to form HCN and aldehyde under alkaline conditions. Although lambda-cyhalothrin is relatively photostable under natural irradiation, with a half-life > 3 wk, its photolysis process is fast under UV irradiation, with a half-life < 10 min. The fate of lambda-cyhalothrin in aquatic ecosystems depends on the nature of system components such as suspended solids (mineral and organic particulates) and aquatic organisms (algae, macrophytes, or aquatic animals). Lambda-cyhalothrin residues dissolved in water decrease rapidly if suspended solids and/or aquatic organisms are present because lambda-cyhalothrin molecules are strongly adsorbed by particulates and plants. Adsorbed lambda-cyhalothrin molecules show decreased degradation rates because they are less accessible to breakdown than free molecules in the water column. On the other hand, lambda-cyhalothrin adsorbed to suspended solids or bottom sediments may provide a mechanism to mitigate its acute toxicity to aquatic organisms by reducing their short-term bioavailability in the water column. The widespread use of lambda-cyhalothrin has resulted in residues in sediment, which have been found to be toxic to aquatic organisms including fish and amphipods. Mitigation measures have been used to reduce the adverse impact of lambda cyhalothrin contributed from agricultural or urban runoff. Mitigation may be achieved by reducing the quantity of runoff and suspended solid content in runoff through wetlands, detention ponds, or vegetated ditches. PMID- 18418955 TI - Lead contamination in Uruguay: the "La Teja" neighborhood case. AB - Lead, ubiquitous in the environment as a result of mining and industrialization, is found as a contaminant in humans although it has no known physiological function there. Lead-exposed children are known to be the population with the highest potential health risks. The recommended biomarker to assess environmental lead exposure in animals is lead level in blood. Before 2001, the Department of Toxicology and Environmental Hygiene was the only team to produce human monitoring data on Uruguayan populations (Manay 2001a,b; Manay et al. 1999). Lead pollution in Uruguay first received official attention during the 2001 La Teja poisoning episode. It was in the La Teja neighbourhood of Montevideo that high BLL were found in children (as high as 20 microg/dL), prompting corrective responses from Health and Environmental authorities. Growing awareness of environmental lead pollution and consequential human health effects from that event, resulted in public debate and demands for solutions from Health and Environmental authorities. Citizens demanded public disclosure of information concerning lead pollution and wanted action to address contaminated Uruguayan sites. In response, the Ministry of Health assembled an interinstitutional multidisciplinary committee, with delegates from health, environmental, labor, educational, and social security authorities, as well as community nongovernmental organizations (NGOs), among others. The University of the Republic was designated to serve as the main responsible entity for technical advice and support. After 2001, new research on lead pollution was undertaken and included multidisciplinary studies with communities in response to health risk alerts. The main emphasis was placed on children exposed to environmental lead. Major sources of Uruguayan lead contamination, similar to those in other developing countries, result from metallurgical industries, lead-acid battery processing, lead wire and pipe factories, metal foundries, metal recyclers, leaded gasoline (before December 2003), lead water pipes in old houses, and scrap and smelter solid wastes, among others. Nonoccupational lead exposure usually results from living in or near current or former manufacturing areas or improper handling of lead-containing materials or solid wastes (a particularly important health risk for children). In this chapter, we reviewed available studies published or reported after the pollution events first announced in 2001. These studies include data on exposure, health, and actions taken to mitigate or prevent lead exposure from pollution events in Uruguay. Uruguay adopted CDC's 10 microg/dL as the reference BLL for children (CDC 1991) and a BLL of 30 microg/dL for workers (from the ACGIH standard). Environmental authorities adopted the Canadian reference concentrations for soil: residential and playgrounds (> 140 mg/kg) or industrial areas (> 600 mg/kg) (CCME 2006). Most studies reviewed addressed soil pollution as the main source of lead exposure. Results of thousands of analyses indicated that most children had BLL above reference intervention limits. A significant decrease in BLL was also found over time in the study results, demonstrating the importance of medical intervention, nutrition, and environmental education. The severity of lead pollution discovered required official governmental actions, both to reduce sources of lead contamination and to address the health implications for children who had been exposed to environmental or industrial lead pollution. Dogs were discovered to be useful sentinels for environmental lead pollution; they had higher BLL than children when exposed to the same polluted environment and developed symptoms of lead intoxication earlier and at lower BLL than did children. This same pattern was also observed in families with children and pet dogs living in the La Teja neighborhood. This discovery renders dogs prospectively useful in lead pollution monitoring and diagnosis, particularly in developing countries. BLL results from similar human lead exposure studies conducted 10 yr apart showed significant BLL reductions, after 10 yr, for nonoccupationally exposed Uruguayans. The phase-out of leaded gasoline is thought to have contributed to this improvement. New laws to address occupational and environmental exposures were passed to prevent new cases of lead contamination, and new research studies are underway to monitor lead pollution. Moreover, a systematic surveillance screening program for lead workers and children is planned, although it is not yet underway. The sensitization of the public to the lead pollution problem has been a key driver of governmental action to mitigate and prevent further lead pollution in Uruguay. The changes made since 2001 appear to have yielded positive results. BLL from different populations studied more recently show decreased lead levels, suggesting a lower contribution of environmental lead to exposure of children and nonoccupationally exposed adults. The diverse analytical data collected on lead pollution in Uruguay between 2001 and 2004 were the main ingredient that allowed effective identification of lead pollution in Uruguay and paved the way for official intervention to prevent new pollution events. Nevertheless, full research studies must still be done, including both spot analysis of environmental soil, air, and water samples, and extensive screening of BLL. Future health and environmental actions are needed, not only to remediate known areas of lead pollution, but also to investigate other sources of potential health risks. PMID- 18418956 TI - Applications of carboxylesterase activity in environmental monitoring and toxicity identification evaluations (TIEs). AB - This review has examined a number of issues surrounding the use of carboxylesterase activity in environmental monitoring. It is clear that carboxylesterases are important enzymes that deserve increased study. This class of enzymes appears to have promise for employment in environmental monitoring with a number of organisms and testing scenarios, and it is appropriate for inclusion in standard monitoring assays. Given the ease of most activity assays, it is logical to report carboxylesterase activity levels as well as other esterases (e.g., acetylcholinesterase). Although it is still unclear as to whether acetylcholinesterase or carboxylesterase is the most "appropriate" biomarker, there are sufficient data to suggest that at the very least further studies should be performed with carboxylesterases. Most likely, data will show that it is optimal to measure activity for both enzymes whenever possible. Acetylcholinesterase has the distinct advantage of a clear biological function, whereas the endogenous role of carboxylesterases is still unclear. However, a combination of activity measurements for the two enzyme systems will provide a much more detailed picture of organism health and insecticide exposure. The main outstanding issues are the choice of substrate for activity assays and which tissues/organisms are most appropriate for monitoring studies. Substrate choice is very important, because carboxylesterase activity consists of multiple isozymes that most likely fluctuate on an organism- and tissue-specific basis. It is therefore difficult to compare work in one organism with a specific substrate with work performed in a different organism with a different substrate. An attempt should therefore be made to standardize the method. The most logical choice is PNPA (p-nitrophenyl acetate), as this substrate is commercially available, requires inexpensive optics for assay measurements, and has been used extensively in the literature. However, none of these beneficial properties indicates that the substrate is an appropriate surrogate for a specific compound, e.g., pyrethroid-hydrolyzing activity. It will most likely be necessary to have more specific surrogate substrates for use in assays that require information on the ability to detoxify/hydrolyze specific environmental contaminants. The use of carboxylesterase activity in TIE protocols appears to have excellent promise, but there are further technical issues that should be addressed to increase the utility of the method. The main concerns include the large amount of nonspecific protein added to the testing system, which can lead to undesirable side effects including nonspecific reductions in observed toxicity, decrease in dissolved oxygen content, and organism growth. It is probable that these issues can be resolved with further assay development. The ideal solution would be to have a commercial recombinant carboxylesterase that possessed elevated pyrethroid hydrolysis activity and which was readily available, homogeneous, and inexpensive. The availability of such an enzyme would address nearly all the current method shortcomings. Such a preparation would be extremely useful for the aquatic toxicology community. Further work should focus on screening available esterases for stability, cost, and activity on pyrethroids, with specific focus on esterases capable of distinguishing type I from type II pyrethroids. It would also be beneficial to identify esterases that are not sensitive to OP insecticides. Many esterases and lipases are available as sets to test chemical reactions for green chemistry, enabling large-scale screening. Other potential approaches to increase the utility of the enzyme include derivatization with polyethylene glycol (PEG) or cyanuric acid chloride to increase stability and reduce microbial degradation. It is also possible that the enzyme could be formulated in a sol gel preparation to increase stability. It is likely that the use of carboxylesterase addition will increase for applications in sediment TIEs. Carboxylesterases are an interesting and useful enzyme family that deserves further study for applications in environmental monitoring as well as to increase our understanding of the fundamental biological role(s) of these enzymes. There are, of course, other enzymes that show high esterase activity on pyrethroids but are not technically carboxylesterases in the alpha/beta-hydrolase fold protein family. These enzymes should also be examined for use in TIE protocols and "esterase" arrays as well as for general applications in environmental monitoring. One can envision the creation of a standardized screen of enzymes with esterase activity to (1) identify environmental contaminants, (2) estimate the potential toxic effects of new compounds on a range of organisms, and (3) monitor organism exposure to agrochemicals (and potentially other contaminants). This approach would provide a multibiomarker integrative assessment of esterase inhibiting potential of a compound or mixture. In conclusion, much is still unknown about this enzyme family, indicating that this area is still wide open to researchers interested in the applications of carboxylesterase activity as well as basic biological questions into the nature of enzyme activity and the endogenous role of the enzyme. PMID- 18418957 TI - [Speech on the establishing meeting of Doctor Society of Integrative Medicine, Chinese Medical Doctor Association]. PMID- 18418958 TI - [Treatment of chronic hepatitis B with integrative medicine]. PMID- 18418959 TI - [Paying attention to TCM and integrative medicine in treating YMDD mutation of chronic hepatitis B]. PMID- 18418960 TI - [Situation and prospect of TCM treatment of chronic hepatitis C]. PMID- 18418961 TI - [Clinical study on treatment of severe hepatitis with removing dampness and purgative method]. AB - OBJECTIVE: To evaluate the clinical effect of removing dampness and purgative (RDP) method in treating acute, subacute and chronic severe hepatitis. METHODS: One hundred and twenty cases of severe hepatitis were randomly assigned to 2 groups, 60 patients in the control group were treated with routine Western medicine, 60 patients in the treatment group were treated with the same Western medicine plus Chinese medicine prescribed based on RDP principle orally and/or via enema. Fourteen days of treatment constituted one therapeutic course, and patients were treated for 3 courses. Changes of clinical symptoms and signs, complication occurrence, liver function, serum markers of hepatitis B virus, and some biological indexes were observed and compared. The case fatality rate was compared after a 6-month follow-up. RESULTS: The total effective rate and marked improving rate in the treatment group was 71.7% (43/60 cases) and 48.3% (29/60 cases) respectively, while those in the control group, 51.7% (31/60 cases) and 20.0% (12/60 cases) respectively, showing significant difference between the two groups (P < 0.05). After treatment, the clinical symptoms and signs were relieved and complications were reduced in the treatment group, showing marked improvement as compared with that in the control group (P < 0.05). ALT, AST, TBil, quantitative titer of HBV-DNA and HBeAg decreased markedly, and ALB, prothrom base activity (PTA) and total cholesterol (TC) increased significantly in both groups after treatment (P < 0.01). Significant difference was found in AST, TBil, PTA and quantitative titer of HBV-DNA between the two groups (P < 0.05, P < 0.01). In the 6-month follow-up, the case fatality rate was 23.3% (14/60 cases) in the treatment group, significantly lower than that in the control group (P < 0.05), which was 41.6% (25/60 case) CONCLUSION: RDP treatment is helpful to improve the prognosis of patients with severe hepatitis, it is one of the effective measures for enhancing the efficacy of comprehensive treatment. PMID- 18418962 TI - [Effects of oxymatrine on serum cytokines and hepatic fibrotic indexes in patients with chronic hepatitis B]. AB - OBJECTIVE: To study the effect of oxymatrine on serum cytokines and hepatic fibrotic indexes in patients with chronic hepatitis B (CHB). METHODS: Eighty-two CHB patients were randomly assigned to the control group treated with Western routine therapy and the treated group treated by Western routine therapy plus oxymatrine. The treatment course was 24 weeks. Another group with 20 healthy subjects was set as the normal control group. The serum levels of transforming growth factor-beta1 (TGF-beta1), interleukin-6 (IL-6), hyaline acid (HA), collagen type IV (C-IV) and laminin (LN) were measured by ELISA and RIA before and after treatment. RESULTS: The serum levels of TGF-beta1, IL-6, HA, C-IV and LN in CHB patients were significantly higher than those in the normal control group (P < 0.01). After 24-week treatment with oxymatrine, these abnormal indexes in the treated group were significantly lowered, as compared with those before treatment and in the control group, the differences were significant (all P < 0.01). CONCLUSION: Oxymatrine could lower the levels of cytokines, including TGF beta1, IL-6, etc. in patients with CHB, which might be one of the mechanisms of its action in reversing liver fibrosis. PMID- 18418963 TI - [Study on TCM syndrome typing of chronic hepatitis B]. AB - OBJECTIVE: To explore the method for TCM syndrome typing of chronic hepatitis B (CHB). METHODS: Clinical questionnaire of epidemiological investigation was established, by which the relative information from 871 patients with CHB was collected on the spot to conduct multi-analysis, including factor analysis, cluster analysis, correspondence analysis, etc. Then the population basic TCM syndrome types and the individual TCM syndrome type of patients were obtained adopting the new combination of factor analysis and variable cluster, and the corresponding relation between TCM syndrome type and the figures of tongue and pulse was analyzed as well. RESULTS: Analysis on distribution of TCM syndrome type in patients showed that single syndrome type presented in 380 cases (43.6%), including Gan-Shen yin deficiency type in 126 (14.5%), Gan-Dan dampness-heat type in 126 (14.5%), Gan-depression and Pi-deficiency type in 128 (14.7%); compound syndrome type presented in 301 (34.6%); the other 190 patients (14.5%) without any symptom was regarded as no syndrome type. Multiple correspondence analysis showed that Gan-Shen yin deficiency type is related to red tongue, thin yellowish fur, and taut pulse; Gan-Dan dampness-heat type is related to yellowish greasy fur and slippery pulse; Gan-depression with Pi-deficiency type is related to thin whitish fur and taut thready pulse. CONCLUSION: The above-mentioned typing of TCM syndrome well coincided with the clinical practice, and the correspondence between single syndrome type and the manifestation of tongue and pulse is obvious. PMID- 18418964 TI - [Applying Fuzheng Huayu Gantang comprehensive therapeutic program for treatment of post-hepatitis B liver cirrhosis complicated with glyco-metabolic abnormality]. AB - OBJECTIVE: To study the clinical effects of Fuzheng Huayu Gantang comprehensive therapeutic program (FHGP) on post-hepatitis B liver cirrhosis associated with glyco-metabolic abnormality (LCGA). METHODS: The patients with LCGA enrolled in the randomized controlled clinical trial were assigned to 2 groups, the treated group (68 cases) and the control group (74 cases), they were treated respectively by FHGP and conventional TCM and Western medicine therapeutic program for 3 months. Indexes including fasting plasma glucose (FPG), 2 h postprandial plasma glucose (2 h PG), fasting insulin (FINS) were detected, homeostasis model assessment insulin resistance (HOMA-IR) was calculated, and the score of syndrome was recorded before and after treatment. Then the effects on syndrome and glyco metabolic abnormality were evaluated through statistical analysis. RESULTS: Level of 2 h PG after treatment was lowered in both groups (P < 0.01), but significant difference was found in the pre-treatment to post-treatment decrement of FPG and HOMA-IR between the two groups (P < 0.05). The syndrome improving rate and the total effective rate on glyco-metabolic abnormality in the treated group were significantly better than those in the control group respectively (85.3% vs 64.9% , P < 0.01; 80.9% vs 62.2%, P < 0.05). CONCLUSION: FHGP has the capability to improve the syndrome and glyco-metabolic abnormality of patients with LCGA. PMID- 18418965 TI - [Removing phlegm and dispelling stasis method combined with Western medicine for treatment of cerebrovascular stenosis]. AB - OBJECTIVE: To investigate the effect of removing phlegm and dispelling stasis method (RPDSM) combined with Western medicine for treatment of cerebrovascular stenosis. METHODS: Seventy enrolled patients were randomly assigned to 3 groups: the Western medicine (WM) group, the integrative medicine (IM) group, and the traditional Chinese medicine (TCM) group. The 21 patients in the WM group were treated with Western medicine as aspirin, Clopidogrel, statins, etc.; the 23 patients in the TCM group were treated with Chinese drugs using the patent preparation Dahuang Zhechong Pill and Tianma Duzhong Capsule as the basic drugs, and supplemented by self-formulated decoctions, selected according to their syndrome types (Tanshi Recipe for dampness-phlegm syndrome, Tanhuo Recipe for fire-phlegm syndrome, Qixu Recipe for qi-deficiency syndrome, and Yang-kang Recipe for yang-excess syndrome); and the 26 patients in the IM group were treated by both TCM and WM with the same drugs and doses mentioned above. The course of treatment was 3 months, and all patients received at least 2 courses in succession. Changes in clinical symptoms and TCM syndrome, levels of C-reactive protein (CRP), platelet aggregation rate (PAR) and fibrinogen (Fib), as well as pictures of medical imaging were observed to evaluate cerebrovascular stenosis. RESULTS: After 6-month treatment, the blood levels of CRP, PAR and Fib were lowered and the number of moderate and severe stenosed vessel lessened in all the three groups (all P < 0.05). The respective total effective rate in the WM, TCM and IM group was 42.9% (9/21 cases), 39.1% (9/23 cases) and 61.5% (16/26 cases), no significant difference was shown among them. CONCLUSION: The integrative Chinese traditional and Western medical treatment for cerebrovascular stenosis shows an increasing trend in improving clinical efficacy and laboratory indexes, combared with pure Western or Chinese medical treatment. PMID- 18418967 TI - [Research on relationship between angiotensinogen gene M235T and TCM syndrome type in essential hypertension patients]. AB - OBJECTIVE: To explore the relationship between angiotensinogen (AGT) gene M235T and TCM syndrome type in essential hypertension (ET) patients. METHODS: The gene mutation frequency of AGT M235T in 168 ET patients and 42 nomotensive (NT) subjects were determined using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique. RESULTS: There was a significant difference in AGT M235T gene mutation between patients of Gan-fire exuberant type and those of yin-yang deficiency type (P < 0.01), homozygote type TT appeared with higher frequency. Multivariate regression linear analysis demonstrated that the genotypes of AGT M235T was correlated with the prognosis of ET to a certain degree. CONCLUSION: Gene mutation of AGT M235T may be associated with the genesis and development of ET, and the TCM syndrome type of ET has its own intrinsic molecular biological background. PMID- 18418966 TI - [Effect of Tongguan capsule on post-intervention patients of coronary heart disease with qi-deficiency and blood stasis syndrome]. AB - OBJECTIVE: To observe the effect of Tongguan Capsule (TGC), a Chinese herbal preparation for supplementing qi and activating blood circulation, in patients after percutaneous coronary intervention (PCI) with qi-deficiency and blood stasis syndrome. METHODS: One hundred patients after successful PCI operation were assigned to two groups, 50 in each group. Western routine therapy with anti thrombosis and anti-coagulant agents were applied in all patients before, during and after operation, while to the patients in the treated group, TGC was given additionally. The therapeutic course for both groups was one month. The efficacy was evaluated by observing the effect on angina pectoris, main TCM syndromes and scores of qi-deficiency syndrome and blood stasis syndrome at 3 time points (the day before, 3 days and 30 days after operation). RESULTS: The total effective rate in relieving angina pectoris was 96.0% (48/50) in the treated group and 92.0% (46/50) in the control group, showing insignificant difference between them (P > 0.05). The score of qi-deficiency in both groups raised 3 days after PCI as compared with that before PCI, but showed no statistical significance (P > 0.05); 30 days after PCI, it increased in the control group, as compared with that before PCI (P < 0.05). The score of blood stasis syndrome significantly lowered at the 3rd and the 30th day after PCI in both groups as compared with before PCI, showing statistical significance (P < 0.05); while in comparing the value at the 3rd day with that at the 30th day, the difference showed significance in the treated group but not in the control group (P < 0.05); and comparison between the two groups at the 30th day showed it was much lower in the treated group (P < 0.05). CONCLUSION: TGC could significantly improve the clinical symptoms of qi deficiency and blood stasis syndrome in patients after PCI. PMID- 18418968 TI - [Effects of Danggui Buxue Decoction on liver fibrosis and hepatic lipid peroxidation in rats]. AB - OBJECTIVE: To observe the effects of Danggui Buxue Decoction (DBD) on liver fibrosis and to explore its mechanism related to hepatic lipid peroxidation in rats. METHODS: Liver fibrosis model was established in 28 rats by the combination of injecting carbon tetrachloride (CCl4) subcutaneously and feeding high lipid and low protein diet. The model rats were randomly divided into 2 groups, the model group (n = 14) and the treated group (n = 14). Besides, a normal group was set up with 10 normal rats. For the treated group, rats were administered with DBD at a dosage of 6 g/kg body weight once a day by gastrogavage starting from the day of modeling for 6 successive weeks and to the control group, equal volume of normal saline was administered instead. The inflammation and fatty degeneration in rat liver tissues were examined with HE staining; the collagen deposition observed with sirius red; the liver function including serum level of alanine transaminase (ALT), aspartate aminotransferase (AST), albumin (Alb) and total bilirubin (TBil) were determined using corresponding test kits; the hepatic lipid peroxidation indexes, including triglyceride (TG), superoxide dismutase (SOD) and malondialdehyde (MDA) were measured by biochemical methods; the hepatic hydroxyproline (Hyp) content was detected with Jamall's method and the expression of collagen type I was analyzed by Western blotting. RESULTS: Compared with those in the normal rats, serum ALT, AST, TBil level, TG and MDA content remarkablely increased, level of Alb and SOD activity decreased, and hepatic fatty degeneration and collagen pathological deposition in liver was more obvious in the model rats (all P < 0.05). While in the DBD group, the hepatic fatty degeneration and collagen deposition were significantly improved, changes of all the above-mentioned indexes were significantly reversed (P <0.05). CONCLUSION: DBD has a good antagonist effect against experimental liver fibrosis, and its mechanism may be related to the anti-lipid peroxidation injury effect. PMID- 18418969 TI - [Effect of Compound Qidan Liquid on early ventricular remodeling after acute myocardial infarction in Chinese mini-pigs]. AB - OBJECTIVE: To investigate the mechanism of Compound Qidan Liquid (CQD) for intervening ventricular remodeling (VR) after acute myocardial infarction (AMI) in Chinese mini-pigs from hemodynamic and collagen metabolic views. METHODS: AMI model of Chinese mini-pigs was established by left anterior descending (LAD) coronary artery ligation. The model pigs were then randomly divided into the sham operative group, the model group, the captopril group, the high and low dose of CQD (hCQD and lCQD) groups, the former two were treated with normal saline and the latter three treated with corresponding drugs by gastrogavage for 4 weeks after modeling. Blood pressure (BP), left ventricular pressure (LVP), maximum ascending velocity of left ventricular pressure (dp/dtmax), myocardial renin (MCR), angiotensin (Ang II), total collagen (TC), procollagen type III (PC III), collagen type IV (CIV), laminin (LM), serum hyaluronic acid (HyA) as well as pathologic changes in myocardium were observed. RESULTS: As compared with in the model group, levels of BP, LVP and dp/dtmax were significantly higher, LM and Ang II were lower in the hCQD group (P < 0.05 or P < 0.01); LVP was higher in the lCQD group (P <0.05); LVP and dp/dtmax were higher, Ang II was lower in the captopril group (P <0.05). Besides, levels of HyA and TC were lower in all the three medicated groups (P <0.01), while the differences of PC III and CIV among groups were insignificant. CONCLUSION: CQD has a beneficial effect in Chinese mini-pigs after AMI for increasing LVP and dp/dtmax, improving myocardial contractility and hemodynamic condition, decreasing myocardial Ang II contents, decreasing deposition of collagen so as to alleviate the pathological process of VR after AMI. PMID- 18418970 TI - [Effect and mechanism of Epimedium flavanoids for aging retardation from viewpoint of transcriptomics and metabonomics]. AB - OBJECTIVE: To investigate the effect and its mechanism of Epimedium flavanoids (EF) in retarding aging with different systematic viewpoints. METHODS: Hypothalamus, pituitary, adrenal and lymphocytes taken from 4-, 10-, 18-, 24 month old rats and from EF treated 24-month old rats were used to measure whole genome mRNA expression by gene array. Serum samples were used for metabonomic assay with high performance liquid chromatography. Using specific gene chip for NF-kappaB signaling pathway to detect the gene expression of the molecule related to that pathway in lymphocytes. Then, a neural network (NN) model was established upon the data obtained to quantitatively evaluate the degree of aging and the efficacy of drug intervention. RESULTS: Gene expression of 199 genes showedan age dependent pattern, most of which were reversed by EF, and the output of NN model showed that EF made the transcriptomics of 24-month old rats to 8-13 months. Seventeen metabolites among the 1,885 peaks detected were identified to have significant age-depending changes, and EF intervention reset the level of metabolites to a younger (18-month) level. The integral level of gene expression for NF-kappaB signaling pathway decreased significantly along with the increasing of age, and was significantly elevated by EF, NN model showed it approached to 10.5-month old. CONCLUSION: Phenotype of aging at different levels demonstrates a common age-dependent trend; EF can reverse this age-dependent change at different levels in a synchronous manner. PMID- 18418971 TI - [Intervening effect of Qiangxin Fumai Granule on arrhythmia induced by right coronary artery ischemia/reperfusion in rabbits]. AB - OBJECTIVE: To observe the development of arrhythmia induced by ischemia/reperfusion (I/R) of the right coronary artery in rabbits and the intervening effect of Chinese medicine Qiangxin Fumai Granule (QFG), a Chinese preparation for activating yang and promoting blood circulation, on it. METHODS: Rabbit right coronary artery I/R model was adopted. Forty healthy adult rabbits were randomly divided into 5 groups, the sham-operation group, the model group, the atropine group, the high-dose QFG group, and the low-dose QFG group, eight in each group. The drugs were administered via duodenal perfusion 10 min after ischemia. The changes of AA interval before and after medication were observed and the scores of arrhythmia were calculated. RESULTS: During ischemia period, AA interval prolonged for more than 40 ms in the model group, and the scores of arrhythmia showed a rising trend along with the prolonging of ischemia, with the presence of atrial-ventricular block (AVB) and aggravating of sinus and atrial arrhythmia; during reperfusion period, the incidence of AVB decreased, and AA interval somewhat decreased. The AA intervals and scores of arrhythmia in the high and low-dose QFG groups were significantly lower than those in the model group respectively (P < 0.01 or P < 0.05). CONCLUSION: QFG is beneficial for shortening AA interval and preventing arrhythmia induced by I/R. PMID- 18418972 TI - [Effects of 3 genosides on bio-thermodynamic expression of splenic lymphocyte in mice]. AB - OBJECTIVE: To establish a new method for screening active ingredients of Chinese herbs by determining different bio-thermodynamic effects of 3 genosides on splenic lymphocyte of mice. METHODS: Using a thermal bioactivity monitoring system, the maximum heat output (mHO), average metabolic heat (MH) and constant of decrease rate (DR) of lymphocyte were determined based on the growth metabolic power-time curve, and the outcomes were verified by MIT. RESULTS: The mHO and MH increased and the DR decreased after lymphocytes being exposed to the 3 genosides in different concentrations, arranged upon their potency as genoside Rg3 > genoside Rg2 > genoside Rg1 (merely insignificant effect). MTT showed the same results. CONCLUSION: Heat activity monitoring system could precisely display the different bio-thermal dynamic effects of 3 genosides on splenic lymphocyte. PMID- 18418973 TI - [Effects of Xiongshao capsule on blood vessel collagenase gene expression in experimental rabbits with arterial restenosis]. AB - OBJECTIVE: To observe the effects of Xiongshao Capsule (XSC) on blood vessel collagenase gene expression in experimental rabbits with arterial restenosis, and to probe its mechanisms for preventing restenosis. METHODS: Restenosis rabbit model was established by injuring endothelium of abdominal aorta by balloon dilation and feeding with high fatty diet for 6 weeks. Eighty rabbits were randomly allocated into 8 groups, Group A, normal rabbit for control; Group B, rabbit with simple injured arterial endothelium; Group C, model rabbits at different times after modeling (3 days for Group C1, 2 weeks for Group C2, and 6 weeks for Group C3); Group D, model rabbit treated with Probucol for 6 weeks; Group E and F, model rabbit treated with small and large dose of XSC respectively. The effect of XSC on collagenase gene expression during the course of restenosis was observed adopting RT-PCR method and computer image analyzer, and its mechanisms in preventing RS were probed by combined analyzing the change of collagen and patho-morphological examination. RESULTS: Compensatory dilation of lumens appeared at the end of the 2nd week; while 6 weeks after modeling, the diameters of lumens obviously diminished with an apparently increased proliferation index. The cell proliferation inhibiting effect in Group D and F was significant. The total amount of collagen increased and reached the peak at the 2nd week but without conspicuous accumulation on intima, which increased gradually and reached its peak at the 6th week. In Group D-F, especially in Group F, the amount of collagen in vascular wall (intima, media and externa) was lesser than that in Groups C. MMP-1 mRNA showed weak expression in Group A and Group C1 C3; significant difference only existed in comparing Group F with C3 (P < 0.05). CONCLUSION: XSC could markedly increase the MMP-1 mRNA expression in injured portion of vessels, suggesting that its action in preventing RS might be related with the up-regulating of MMP-1 mRNA expression, increasing collagen degradation and reducing collagen deposition in vascular wall. PMID- 18418974 TI - [Effect of Herba Epimedil Brevicornus and prepared Radix Rehmannia on glucocorticoid receptor in glucocorticoid receptor down-regulated rats]. AB - OBJECTIVE: To evaluate the effect of Herba Epimedii Brevicornus (HEB) and prepared Radix Rehmannia (RR), the representative Chinese herbs for warming yang and nourishing yin on glucocorticoid receptor (GR) in GR down-regulated rats. METHODS: The GR down-regulated model was established by subcutaneous injection of hydrocortisone. Seventy-two SD male rats were randomly divided into four groups: the normal group; the model group (MG) ; the HEB group, and the RR group. And every group was subdivided to 3 batches depending on the time points (the 3rd, 7th and 14th day after modeling) of observation, with 6 rats in each batch. Changes in blood level of corticosterone (GS), as well as protein expression and binding power of GR in splenic lymphocytes at corresponding time points of the batches were determined by RIA and flow cytometry. RESULTS: No significantly difference was found in blood GS levels of the model group at all the time points, as compared with that of the normal group (P > 0.05), while that in the RR group was significantly lowered at the 14th day, and showed significant difference from that in the model group (P < 0.05). The protein expression and binding power in the model rats were lower than those in normal ones (all P < 0.01), but they were significantly higher on the 7th and 14th day in the RR group (P < 0.01), and on the 14th day in the HEB group higher than those in the model group, respectively (all P < 0.01). CONCLUSION: Both HEB and RR, in dosage-form of water decoction, could up-regulate the protein expression and binding power of GR in GR down-regulated model rats, but different in acting time. PMID- 18418975 TI - [Effect of extract of Ginkgo biloba on doxorubicin-associated cardiotoxicity in patients with breast cancer]. AB - OBJECTIVE: To study the effect of extract of Ginkgo biloba (Egb761) on doxorubicin-associated cardiotoxicity in patients with breast cancer (BC). METHODS: Sixty BC patients in stage IV were randomly assigned to two groups, the control group was treated with chemotherapy, using 4 cycles of PA protocol alone and the treated group with the same chemotherapy and Egb761. Changes in electrocardiogram (ECG), myocardial enzyme spectrum (MES) and ultrasono cardiogram (USCG) before and after treatment were observed. RESULTS: After treatment, the incidence of abnormal ECG was lower in the treated group than in the control group (6.7% vs 30.0%); significant differences were found between the two groups in the parameters of MES (P< 0.05); USCG showed significant difference between the two groups in left ventricular diastolic diameter (LVDd), left ventricular systolic diameter (LVDs), ratio of early and late diastolic transmitral peak flow velocity (E/A) and fractional shortening (FS), while there was no significant difference in ejection fraction (EF). CONCLUSION: Egb761 is an ideal drug for preventing and reducing the acute doxorbincin-induced cardiotoxicity; it could also be helpful for alleviating the chronic cardiotoxicity. PMID- 18418976 TI - [Effect of Yishen Tiaojing Recipe on hypogonadism induced by Tripterygium glycosides in women patients with nephropathy]. AB - OBJECTIVE: To explore the effect of Yishen Tiaojing Recipe (YSTJR) on sex gland and kidney in nephropathy patients with hypogonadism induced by Tripterygium glycosides (TG). METHODS: Fifty-eight women patients with chronic glomerular disease were assigned to two groups, the 34 patients in the treated group were given TP 20 mg three times a day plus YSTJR, one dose daily, while the 24 in the control group were given TP 20 mg three times a day alone. The course of treatment lasted for 3 months. Serum levels of estradiol (E2), follicle stimulating hormone (FSH), and luteotrophic hormone (LH), as well as urinary protein and red blood cell (RBC) count in urine were determined before and after treatment. RESULTS: Level of E2 decreased, FSH and LH increased in the control group after treatment (P < 0.05), while those indexes unchanged in the treated group. Quantity of urinary protein decreased in both two groups after treatment (P < 0.05), but the decrement was more significant in the treated group than in the control group (P < 0.05). Differences of all the above-mentioned indexes between the two groups after treatment were significant (all P < 0.05). CONCLUSION: YSTJR could prevent the injury of TP on sex gland in women patients with nephropathy, and is benificial for the treatment of primary chronic renal glomerular diseases. PMID- 18418977 TI - [Combination of effective components, the new pattern of drugs derived from modern Chinese herbal medicine]. AB - The thinking of research on developing Chinese herbal medicine (CHM) has turned from the traditional exploration for active ingredients to develop microcosmic compound drug based on TCM and/or modern medical theories, which leads to the emerge of combination of effective components (CEC), a new pattern of developing modern Chinese herbal medicine. Meanwhile its researching targets and methods, including objectivization of treated diseases, optimization of compatibility proportion, standardization of component types, definition of synergistic and antagonistic effects, and minimization of toxic and side effects, have also been renewed. The existent problems in need of improvement were put forward on the basis of current researching situation of CEC pattern in order to construct new drugs, enrich the theoretical connotation of TCM and promote the process of TCM modernization. PMID- 18418978 TI - [Sub-health state and chronic fatigue syndrome]. AB - This paper points out that the sub-health state is not equal to chronic fatigue syndrome (CFS) on basis of elaborating the concept and category of sub-health. And the present understanding on concepts of fatigue, chronic fatigue and CFS, as well as the diagnosis criteria and differential diagnosis of CFS are discussed systematically. PMID- 18418979 TI - [Launching an epidemiologic investigation on characteristics of syndrome for studying critical syndrome types]. AB - Study on syndrome type should be combined with clinic closely to make clear the regularities of distribution and development of syndrome types in diseases, as well as the statues of different syndromes in the genesis and development processes of diseases. Aiming at this issue, the authors bring forward the concept of "critical syndrome types", and suggest that for resolving it, launching an epidemiologic investigation on characteristics of syndrome types is necessary. PMID- 18418980 TI - [Discussion on English translation of TCM gynecologic terms]. AB - The existent problems, principles that should be observed and methods for English translation of TCM gynecological terms were discussed, and a scheme of translation for 150 TCM gynecological terms was explored. PMID- 18418981 TI - [Experimental advance of applying emodin for prevention and treatment of liver diseases]. AB - The experimental researches of applying emodin for prevention and treatment of liver diseases in recent years were reviewed. Emodin can inhibit the growth of liver tumor cells in vitro and in vivo, inducing cell apoptosis is one of its mechanisms. Emodin also has the effects of liver protection, anti-liver fibrosis, and so on, the mechanisms for those effects still need more studies. PMID- 18418982 TI - Homelessness and hunger. AB - We employ data from the National Survey of Homeless Assistance Providers and Clients to examine the character and correlates of hunger among homeless people. Our analysis, couched in an adaptation framework, finds more support for the differentiation hypothesis than for the leveling hypothesis: Complex patterns of food insecurity exist at the individual level, and they vary with the resources available (e.g., higher monthly income, regular shelter use) and obstacles faced (e.g., alcohol, drug, and physical and mental health problems). The chronically homeless, who suffer from multiple deficits, appear particularly food-insecure, a finding that favors the desperation hypothesis over its street-wisdom alternative. We conclude that hunger is not uniformly experienced by members of the homeless population. Rather, some individuals are better situated than others to cope with the stressful nature of homelessness when addressing their sustenance needs. PMID- 18418983 TI - Electronic support groups, patient-consumers, and medicalization: the case of contested illness. AB - This article illustrates the role electronic support groups play in consumer driven medicalization. The analysis is based on an observational study of a year in the life of an electronic support group for sufferers of the contested illness fibromyalgia syndrome. The analysis builds on and extends scholarship concerning the growing influence of lay expertise in the context of medical uncertainty by showing how the dominant beliefs and routine practices of this electronic community simultaneously (and paradoxically) challenge the expertise of physicians and encourage the expansion of medicine's jurisdiction. Drawing on their shared embodied expertise, participants confirm the medical character of their problem and its remedy, and they empower each other to search for physicians who will recognize and treat their condition accordingly. Physician compliance is introduced as a useful concept for understanding the relationship between lay expertise, patient-consumer demand, and contemporary (and future) instances of medicalization. PMID- 18418984 TI - Parental health and children's economic well-being. AB - The life course perspective emphasizes that past economic experiences and stage in the life course influence a family's ability to cope with negative life events such as poor health. However, traditional analytic approaches are not well-suited to examine how the impact of negative life events differs based on a family's past economic experiences, nor do they typically account for the potentially spurious association between negative life events and family economic well-being. We use finite mixture modeling to examine how changes in parental health affect children's exposure to poverty. We find that for some children the association between family head's health and children's exposure to poverty is spurious, while for other children family head's poor health is associated with increased risk of economic deprivation. The extent to which a family head's poor health alters children's economic well-being depends on a child's family's underlying economic trajectory and past history of exposure to disadvantage. PMID- 18418985 TI - Incarceration as exposure: the prison, infectious disease, and other stress related illnesses. AB - This article examines the relationship between incarceration and health functioning. Using data from the National Longitudinal Survey of Youth, the relationship between incarceration and more than 20 different measures of health are tested. Using multiple analytic procedures, a distinctive pattern of association emerges. Individuals with a history of incarceration appear consistently more likely to be afflicted with infectious disease and other illnesses associated with stress. In contrast, no consistent relationships were observed between incarceration status and ailments unrelated to stress or infectious disease. The results suggest that exposure to infectious disease and stress are important to understanding the lasting impact of incarceration on health. PMID- 18418986 TI - The resources that matter: fundamental social causes of health disparities and the challenge of intelligence. AB - A robust and very persistent association between indicators of socioeconomic status (SES) and the onset of life-threatening disease is a prominent concern of medical sociology. The persistence of the association over time and its generality across very different places suggests that no fixed set of intervening risk and protective factors can account for the connection. Instead, fundamental cause theory views SES-related resources of knowledge, money, power prestige, and beneficial social connections as flexible resources that allow people to avoid risks and adopt protective strategies no matter what the risk and protective factors are in a given place or time. Recently, however, intelligence has been proposed as an alternative flexible resource that could fully account for the association between SES and health and thereby find its place as the epidemiologists' "elusive fundamental cause" (Gottfredson 2004). We examine the direct effects of intelligence test scores and adult SES in two data sets containing measures of intelligence, SES, and health. In analyses of prospective data from both the Wisconsin Longitudinal Study and the Health and Retirement Survey, we find little evidence of a direct effect of intelligence on health once adult education and income are held constant. In contrast, the significant effects of education and income on health change very little when intelligence is controlled. Although data limitations do not allow a definitive resolution of the issue, this evidence is inconsistent with the claim that intelligence is the elusive fundamental cause of health disparities, and instead supports the idea that the flexible resources people actively use to gain a health advantage are the SES-related resources of knowledge, money, power, prestige, and beneficial social connections. PMID- 18418987 TI - Effects of goal-striving stress on the mental health of black Americans. AB - Although many scholars have theorized about how responding to the stress of blocked opportunities can affect the well-being of black Americans, few scholars have empirically examined the relationships between striving efforts, personal goals, and mental health among black Americans. This investigation examines the relationship between goal-striving stress and mental health in a national sample of black Americans. Results indicate that goal-striving stress is significantly related to lower levels of happiness, life satisfaction, self-esteem, and higher levels of psychological distress. We find that poverty status moderates the relationship between goal-striving stress and mental health. Compared to poorer persons, individuals above poverty with high goal-striving stress have significantly lower levels of happiness and life satisfaction. Overall, the findings provide a more complete context for understanding associations among socioeconomic status, goal-striving stress, and adverse mental health outcomes among black Americans. PMID- 18418988 TI - Cross-national comparison of social support structures between Taiwan and the United States. AB - The study compares the effects of structural bases and functional elements of social support on mental health in Taiwan and the United States, using the study conducted in the United States by Lin, Ye, and Ensel (1999) as a reference. Based on a nationally representative sample of Taiwanese adults (n = 2,835), a fundamental similarity in social support structure and function between the two countries was observed. First, the structural bases of social support had a hierarchical order in their effects on depression: Binding (presence of an intimate relationship) was the strongest in reducing depression, whereas belonging (community participation) was the weakest, with bonding (social networks) in between. Regarding the functional elements, perceived social support was a better protector of mental health than actual social support, a finding in line with previous research. On the other hand, several notable differences in the structural bases and functional elements of social support between the two societies were observed, possibly due to the differential cultural and historical characteristics. PMID- 18418989 TI - On hastening death without violating legal and moral prohibitions. PMID- 18418990 TI - Clinical anatomy of the canine brain using magnetic resonance imaging. AB - The purpose of this study was to produce an magnetic resonsnce (MR) image atlas of clinically relevant brain anatomy and to relate this neuroanatomy to clinical signs. The brain of a large mixed breed dog was imaged in transverse, sagittal, and dorsal planes using a 1.5 T MR unit and the following pulse sequences: Turbo (fast) spin echo (TSE) T2, T1, and T2- weighted spatial and chemical shift encoded excitation sequence. Relevant neuroanatomic structures were identified using anatomic texts, sectioned cadaver heads, and previously published atlases. Major subdivisions of the brain were mapped and the neurologic signs of lesions in these divisions were described. TSE T2-weighted images were found to be the most useful for identifying clinically relevant neuroanatomy. Relating clinical signs to morphology as seen on MR will assist veterinarians to better understand clinically relevant neuroanatomy in MR images. PMID- 18418991 TI - Objective radiographic assessment of abdominal sand accumulation in horses. AB - Gastrointestinal sand accumulation is a common cause of equine colic. Subjective assessment of sand accumulation on abdominal radiographs has been used as a diagnostic aid; however, there is poor correlation between clinical, diagnostic, and surgical findings. The purpose of this study was to develop an objective method of assessing radiographic sand accumulation in order to improve the diagnostic utility of radiography for sand colic. Fifty-one equine abdominal radiographic examinations were reviewed, with approximately half of the imaged patients having a clinical diagnosis of sand colic. Initially, four observers independently reviewed these radiographic studies to subjectively decide whether or not sand accumulation was sufficient to cause colic. Subsequently, an objective scoring system was developed using various radiographic parameters that yielded a score ranging from one to 12 for quantity and apparent density of sand accumulation. Inter- and intraobserver results using the subjective scoring method had significant differences among all observers. Subjective assessment was also deemed to be inaccurate for colic prediction. Using the objective scoring system, there were no significant differences between or within the observers' results. A score of seven out of 12 was found to have an 83% likelihood of being associated with a positive diagnosis of sand colic. The designed objective scoring method creates a more uniform and accurate method to assess the sand accumulation. PMID- 18418992 TI - Factors affecting width of the canine femorotibial joint space in nonweight bearing radiographs. AB - Caudocranial stifle radiographs with variations in positioning were made in two greyhound cadavers. Radiographs were repeated after each of three interventions: cranial cruciate desmotomy; release of the caudal horn of the medial meniscus; complete medial meniscectomy. The joint space on medial and lateral aspects of the joint was measured by a observer who was unaware of positioning or intervention. One dog had significantly wider joint space than the other (1.0 vs. 1.5mm). The lateral aspect of the joint space was wider than the medial aspect (1.7 vs. 0.7 mm). Medial rotation of the stifle resulted in an increase of 0.4 mm in width of the lateral joint space, whereas lateral rotation of the stifle reduced the lateral joint space by an average of 0.4 mm. Decentering the X-ray beam had no significant effect on joint space width. Tension increased the width of the medial joint space by an average of 1.2 mm and the lateral aspect by an average of 1.3 mm. Cranial cruciate desmotomy resulted in an average 0.3 mm increase in width of the joint space, and medial meniscectomy with an average 0.2 mm reduction of the joint space. Although the femorotibial joint space was affected by iatrogenic stifle injuries and by medial or lateral rotation, these changes were less than the differences between the two dogs. Hence it seems unlikely that the small changes in joint space width associated with cruciate ligament desmotomy and medial meniscectomy will be detectable in clinical practice. PMID- 18418993 TI - Magnetic resonance imaging features of spinal epidural empyema in five dogs. AB - Spinal epidural empyema is defined an accumulation of purulent material in the epidural space of the vertebral canal. Spinal epidural empyema should be considered as a differential diagnosis in dogs with pyrexia, spinal pain, and rapidly progressing myelopathy. Magnetic resonance (MR) imaging is the imaging test of choice in humans. Here, we describe the MR imaging features of five dogs with confirmed spinal epidural empyema. The epidural lesions appeared as high or mixed signal masses in T2-weighted (T2W) images. Increased signal within the spinal cord gray matter at the site of the lesion was detected in T2W images in all dogs. Two patterns of enhancement were detected on postcontrast T1-weighted (T1W) images. Mild to moderate peripheral enhancement was seen in three dogs and a diffuse pattern of enhancement was seen in one. Discospondylitis was identified in three dogs on T1W postcontrast images. Decompressive spinal surgery was performed in all dogs. Bacteria isolated from the abnormal epidural tissue were Enterobacter cloacae, coagulase-positive Staphylococci, Pasteurella multocida, and Escherichia coli. In one dog bacteria were not isolated. These MR imaging features, along with appropriate clinical signs, can allow prompt diagnosis and appropriate treatment planning. PMID- 18418994 TI - Dual-phase computed tomographic angiography in three dogs with pancreatic insulinoma. AB - This article describes the findings in three dogs with histopathologically confirmed pancreatic insulinoma using dual-phase computed tomographic angiography (CTA). In all three dogs, dual-phase CTA findings identified lesions not seen on ultrasonography, including the actual identification of the primary pancreatic neoplasm in two dogs. CTA findings were in agreement with the surgical and histopathological findings. In two dogs, the insulinomas were found to have a strong enhancement during the arterial phase of the study but not at the other phases, which stresses the importance of dual-phase computed tomography for the diagnosis of this type of pancreatic neoplasia, in agreement with current knowledge in humans. PMID- 18418995 TI - Imaging diagnosis--polioencephalomalacia in a calf. AB - A 2-month-old mix-breed calf developed acute blindness and ataxia. Serum thiamine concentration was deficient. In antemortem magnetic resonance imaging there were laminar T2-hyperintense regions extending along the cerebral cortex that primarily affected the gray matter. The lesions were relatively symmetric between the left and right hemispheres but no abnormalities were present at the frontal lobes. At necropsy, laminar autofluorescence of the cerebral cortex was observed under ultraviolet exposure at 365 nm, consistent with a diagnosis of polioencephalomalacia. Polioencephalomalacia in the bovine species is compared with that in other species, namely humans, dogs, and cats. PMID- 18418996 TI - Imaging diagnosis--spinal injury following aberrant microchip implantation. PMID- 18418997 TI - Use of contrast-enhanced ultrasound for characterization of focal splenic lesions. AB - Contrast-enhanced ultrasound was used to study focal and multifocal lesions of the spleen in 26 dogs and two cats affected by 11 benign and 18 malignant splenic diseases. A second-generation microbubble contrast medium (Sonovue) was injected into the cephalic vein and enhancement patterns were subjectively described and time intensity curves calculated. Final diagnosis was obtained by histopathologic examination after splenectomy (n=19) or by needle aspiration and sonographic follow-up after 4 and 8 weeks (n=9). Contrast-enhanced ultrasound parameters, improving the characterization between benign and malignant lesions, were established. The most useful criterion was the hypoechogenicity of the lesion in the wash-out phase combined with the presence of tortuous feeding vessels, which was observed in association with malignancy. All malignant lesions were hypoechoic to the surrounding spleen 30s after starting the contrast medium injection. Lymphosarcoma and hemangiosarcoma had characteristic perfusion patterns. Lymphosarcoma had rapid time to peak and early wash-out phase with a honeycomb pattern during the wash-out. Hemangiosarcomas were large nonperfused masses in all phases surrounded by hypervascular splenic parenchyma. Benign lesions except one hematoma and a benign histiocytoma had the same perfusion pattern as the surrounding spleen. Ultrasonographic and contrast-enhanced ultrasound findings of an accessory spleen are reported. Contrast-enhanced ultrasound can improve the characterization of focal or multifocal lesions of the spleen. PMID- 18418998 TI - Ultrasonographic evaluation of vitreous degeneration in normal dogs. AB - Vitreous degeneration is common in dogs and may be associated with cataract formation. Vitreous degeneration may be identified using B-mode ultrasonography and appears as multiple, small, motile, point-like echoes within the vitreous cavity. In humans, vitreous degeneration has also been observed in normal aging eyes but the incidence of vitreous degeneration in dogs without cataract has not previously been documented. The purpose of this study was to describe the ultrasonographic appearance of vitreous degeneration and to investigate its incidence in a population of dogs without cataract or other apparent eye disease. The eyes of 62 dogs were evaluated as part of a prospective study. All dogs underwent ophthalmological and ultrasonographic examinations and vitreal changes were graded on ultrasonography using a predetermined grading scheme. Vitreous degeneration was found in 20% (23/114) of the eyes on ultrasonographic examination but in only 8% (9/114) of eyes on direct ophthalmoscopy. Sensitivity and specificity of ophthalmoscopy using ultrasonography as a gold standard were respectively, 39% and 100%. Vitreal syneresis and asteroid hyalosis could be distinguished according to their ultrasonographic characteristics. The probability of having vitreous degeneration increased with the age of the dog (odds ratio = 6.7 for dogs of 7 + years compared with 0-6 years) and also increased in females compared with males (odds ratio = 3.6). Vitreous degeneration, especially mild vitreal syneresis, is not uncommon in normal dogs; it was shown to be an age-related condition and its significance should not be overinterpreted on ocular ultrasonography. PMID- 18418999 TI - Ultrasonographic anatomy of the bovine eye. AB - The purposes of the study were to describe the ultrasonographic appearance and measurements of the normal bovine eye, to compare the measurements to those reported previously for cadaveric eyes and to describe differences between ocular dimensions of Holstein Friesian and Jersey cattle. Sixty transpalpebral ocular ultrasonographic examinations were performed on 30 adult Holstein Friesian cows, and 16 examinations were performed on 8 adult Jersey cows. Transpalpebral ultrasonographic images were obtained with a 10 MHz linear transducer in both horizontal and vertical imaging planes. The ultrasonographic appearance of structures within the bovine eye is similar to that in other species, although the ciliary artery was frequently identified, appearing as a 0.33 +/- 0.04 cm diameter hypoechoic area. The axial length of the globe was significantly greater in Holstein Friesian cattle (3.46 +/- 0.09 cm) compared with Jersey cattle (3.27 +/- 0.19 cm; P = 0.001), although the vitreous depth was smaller in Holstein Friesian cattle (1.46 +/- 0.09 cm) (P = 0.0009). The anterioposterior depth of the lens was significantly greater in Jersey cattle (1.92 +/- 0.11 cm) and the cornea was thinner in Jersey cattle (0.17 +/- 0.02 cm). The appearance and ocular distances for live animals were similar to those reported previously for cadaveric specimens. The knowledge of normal ocular dimensions facilitates the use of ultrasonography in the evaluation of ocular disease in cattle. PMID- 18419000 TI - Imaging diagnosis--sublumbar pseudomycetoma in a Persian cat. AB - A 6-year-old Persian cat was examined for constipation, anorexia, and vomiting that was subsequently found to be due to a pseudomycetoma originating from the descending colon and sublumbar region, and causing mechanical obstruction of the colon and rectum. Multiple discrete hyperechoic foci likely representing fungal grains within the lesion gave the mass a coarse echotexture on ultrasound and was supportive of the diagnosis and computed tomography allowed delineation the extent of the mass. A pseudomycetoma is a granulomatous/pyogranulomatous reaction that surrounds dermatophytic fungal hyphae. Definitive diagnosis of a dermatophytic pseudomycetoma requires identification of the etiologic agent by cultivation or immunohistochemical staining. A pseudomycetoma should be included in the differential diagnosis for an abdominal mass in a Persian cat, especially is accompanied by the sonographic findings noted above. PMID- 18419001 TI - Imaging diagnosis---jejunal vascular malformation in a dog. PMID- 18419002 TI - Thoracic and abdominal organ uptake of 2-deoxy-2-[18F]fluoro-D-glucose (18FDG) with positron emission tomography in the normal dog. AB - Positron emission tomography (PET) has found widespread application for staging and monitoring neoplastic diseases in humans. PET is becoming more available in veterinary medicine, therefore biodistribution of 2-deoxy-2-[18F]fluoro-D-glucose (18FDG) in normal dogs is needed for lesion interpretation in disease states. A large field-of-view (FOV) PET scanner with a 70 cm bore diameter and a 53-cm FOV was used in this study to acquire dynamic 18FDG uptake data from parenchymal organs in seven normal dogs. A 2-h, dynamic list-mode acquisition was initiated simultaneously with intravenous 18FDG injection. Regions of interest (ROIs) were manually drawn over liver, spleen, left and right renal cortices, left ventricular free wall, and thymus. Standardized uptake values (SUVs) of these organs were calculated for 24 5-min frames over the 2-h acquisition. This SUV data from parenchymal organs of normal dogs compares favorably with those of normal humans and will be used in ongoing canine studies using PET to evaluate various diseases. PMID- 18419003 TI - Osteoradionecrosis and radiation induced bone tumors following orthovoltage radiation therapy in dogs. AB - Osteoradionecrosis and radiation-induced bone tumors are rare complications of radiation therapy. Little information regarding these complications is available in veterinary medicine. We characterized these complications and investigated risk factors in 119 dogs (122 sites) that received definitive orthovoltage radiation therapy to appendicular sites. Long-term survival was expected in all dogs. The complications of interest were osteoradionecrosis and secondary bone tumor, evaluated radiographically, histopathologically, or both. Complication rates were estimated using the Kaplan-Meier product-limit method, and Fisher's exact test or chi-square test was used to compare the complication rate. The median survival time was 1405 days, with median follow-up duration of 657 days. There were 10 radiation-induced bone tumors and five radiation-induced fractures, with two dogs developing both, for an overall complication rate of 11%. The latent period ranged from 1.2 to 6.4 years for osteoradionecrosis and from 2.6 to 8.7 years for radiation-induced bone tumor. Complications were significantly higher in the humerus (P < 0.0001), and in dogs younger than 7 years (P = 0.014). Similar assessment of complications in dogs irradiated with megavoltage photons or electrons are needed. PMID- 18419004 TI - Statistical briefing: odds and odds ratios. PMID- 18419005 TI - Magnetic resonance imaging of acquired trigeminal nerve disorders in six dogs. PMID- 18419006 TI - Anaerobic treatment of farm waste: opportunities and challenges. PMID- 18419007 TI - Operation of the SHARON denitrification process to treat sludge reject water using hydrolyzed primary sludge to denitrify. AB - An efficient biological treatment to treat reject water from anaerobic digestion of wastewater sludge is the SHARON denitrification process, which takes place in a chemostat reactor, where aerobic/anoxic periods are alternated under specific hydraulic retention time (HRT) and temperature conditions that favor ammonium oxidizers growth and ensure the total washout of nitrite oxidizers, achieving the biological nitrogen removal over nitrite. An optimized performance of this process to treat Spanish reject water was obtained using methanol and working at an HRT of 2 days, 33 degrees C, and cycle length of 2 hours. Supernatant of hydrolyzed primary sludge was tested to denitrify. Because biochemical oxygen demand was not extremely high in the primary sludge, the fluid dynamics of the system were changed, with respect to the strategy with methanol, but maintaining the reject water influent flowrate. The use of hydrolyzed primary sludge improved the process efficiency, because the alkalinity present in the primary sludge buffered the process until an optimum pH range. PMID- 18419008 TI - Sanitation assessment of wastewater treated by stabilization ponds for potential reuse in agricultural irrigation sanitation assessment. AB - Wastewater reuse has become an important alternative to agricultural irrigation; on the other hand, it poses concern with regard to public health. Total coliform and Escherichia coli concentration, presence of helminth eggs and Salmonella, and physical-chemical parameters were evaluated in raw and treated wastewater. Chemical and biochemical oxygen demand removal efficiency was 74.6 and 77.9%, respectively. As for organic nitrogen, total phosphorus, and total suspended solids, total efficiency removal was 17.4, 12.5, and 32.9%, respectively. The average density of total coliforms and E. coli was 3.5 x 10(9) and 1.8 x 10(8) MPN/100 mL and 1.1 x 10(7) MPN/100 mL and 3.9 x 10(5) MPN/100 mL for raw and treated wastewater, respectively. Ascaris eggs were observed in 80.8% of the samples collected, and viable eggs in 42.3% of the samples. Salmonella was detected in 36.4% of the samples. The values observed in treated wastewater did not show the adequate bacteriological quality, as recommended by World Health Organization (Geneva, Switzerland). Therefore, additional measures should be taken to achieve an improved microbiological and parasitological quality. PMID- 18419009 TI - The anaerobic biodegradability of municipal sludge and fat, oil, and grease at mesophilic conditions. AB - The anaerobic biodegradability of municipal primary and secondary sludge with increasing levels of partially dewatered fat, oil, and grease (FOG) was assessed using a mixed methanogenic culture at 35 "C. Under batch conditions with an acclimated and enriched microbial population, the sludge loading was 3 kg volatile solids/m3 and the highest FOG loading tested was 1.5 kg volatile solids/m3, resulting in a methane yield of 245 mL methane/g sludge volatile solids added at 35 degrees C and 1010 mL methane/g FOG volatile solids added at 35 degrees C. Under semicontinuous feeding conditions, the sludge and sludge plus FOG loading tested were 3 and 3.75 kg volatile solids/m3-d, respectively. Within 23 days of operation, the volatile fatty acid concentrations were reduced below 200 mg chemical oxygen demand/L (187 mg/L as acetic acid). Enhancement of sludge digestion was observed in those reactors where codigestion of sludge and FOG took place, which was attributed to a higher level of microbial activity maintained in these reactors as a result of FOG degradation. The results of this study demonstrate that beneficial use of FOG through codigestion with municipal sludge is feasible. PMID- 18419010 TI - Investigation of nitrification and nitrogen removal from centrate in a submerged attached-growth bioreactor. AB - The purpose of this study was to investigate the nitrification and nitrogen removal from centrate produced in the dewatering process of anaerobically digested sludge, using a single-unit, single-zone submerged attached-growth bioreactor. The nitrogen loading varied from 0.54 to 1.51 kg-N/m3 x d. Stable ammonia oxidation (nitritification) to nitrite was demonstrated. A nitritification efficiency of 98% was achieved, while the denitrification efficiency varied from 84 to 99% (with methanol). The average total nitrogen removal was 85%. Inhibition of nitrite oxidation by a limited penetration of dissolved oxygen into the biofilm and free ammonia resulted in the accumulation of nitrite, while inhibition of ammonia oxidation by free nitrous acid did not occur. The quantity of biomass, in terms of volatile solids, ranged from 10123 to 16034 mg-VS/L of media. PMID- 18419011 TI - Municipal anaerobic digesters for codigestion, energy recovery, and greenhouse gas reductions. AB - Codigestion of five wastes and municipal wastewater sludge was evaluated using full-scale testing. Synergistic, antagonistic, and neutral outcomes were observed depending on codigestate identity and concentration, highlighting the value of careful blending. Yeast waste resulted in notable synergism, increasing biogas production by over 50%, whereas aircraft deicing waste resulted in antagonism at high loadings and neutral outcomes at lower loadings. Restaurant waste codigestion resulted in neutral outcomes. The synergisim with yeast codigestates may have resulted from trace nutrients (i.e., iron, nickel, and cobalt) in the wastes that increased microbiological activity. Antagonist outcomes with deicing waste may have been the result of organic overload or inhibitory deicer constituents. Codigestion of wastes at the feed rates investigated was estimated to produce 0.50 ML/d of methane having an energy equivalent of 17 500 MJ/d. This was estimated to reduce net carbon dioxide emissions by 560 tonnes/y. PMID- 18419012 TI - Phosphorus removal from municipal wastewater by hydrous ferric oxide reactive filtration and coupled chemically enhanced secondary treatment: part I- performance. AB - This work examines the performance of a hydrous ferric oxide (HFO) reactive filtration (RF) process with coupled chemically enhanced secondary treatment (RECYCLE) for phosphorus removal from municipal wastewater (HFO-RF-RECYCLE). A 3 month, 0.95-ML/d (0.25-mgd) demonstration of HFO-RF-RECYCLE was performed at a municipal wastewater treatment plant equipped with oxidation ditches and secondary clarifiers. Influent to the plant averaged 6.0 mg/L phosphorus, with a tertiary effluent average of 0.011 mg/L phosphorus. Iron doses to the plant were low, at 5 mg/L. Inline recycling of HFO solution rejects to the plant influent resulted in a maximum 90.3%, dose-dependent reduction of phosphorus in the secondary effluent at 4.5 ML/d (1.2 mgd). Other results included reduction of total suspended solids and turbidity. A mass balance analysis was performed. We conclude that HFO-RF-RECYCLE may allow very low levels of phosphorus discharge from municipal wastewater treatment plants with a ferric-iron-based tertiary filtration process and residual recycling. PMID- 18419013 TI - Phosphorus removal from municipal wastewater by hydrous ferric oxide reactive filtration and coupled chemically enhanced secondary treatment: part II- mechanism. AB - The removal mechanism of a hydrous ferric oxide (HFO) reactive filtration (RF) process with coupled chemically enhanced secondary treatment (RECYCLE) for phosphorus removal from municipal wastewater (HFO-RF-RECYCLE) was examined. A 0.95-ML/d (0.25-mgd) demonstration of HFO-RF-RECYCLE was performed at a municipal wastewater treatment plant equipped with oxidation ditches and secondary clarifiers. Influent to the plant averaged 6.0 mg/L phosphorus, with a 3-month tertiary effluent average of 0.011 mg/L phosphorus. In addition to aqueous geochemical modeling, experiments with surface charge, scanning electron microscopy, adsorptive capacity, thermal desorption, and most probable number of iron(III)-reducing bacteria were performed on samples from the system, to determine the major phosphorus-removal pathways. Results suggest that, in addition to filtration of particulate phosphorus, the low tertiary effluent total phosphorus result was achieved by adsorption. PMID- 18419015 TI - Implications of urine-to-feces ratio in the thermophilic anaerobic digestion of swine waste. AB - Thermophilic anaerobic digestion of swine manure represents a potential waste treatment technology to address environmental concerns, such as odor emissions and removal of pathogenic microorganisms. However, there are concerns relative to the stability of this process when swine manure is the sole substrate. In this study, the potential of biogas production from swine manure as the sole substrate under thermophilic (50 degrees C) conditions was investigated in the laboratory, to determine whether separation of urine and feces as part of the waste collection process would benefit anaerobic digestion. Effluent from a continuously stirred tank reactor was used as the inoculum for batch tests, in which the substrate contained three different concentrations of urine (urine free, as-excreted urine-to-feces ratio and double the as-excreted urine-to-feces ratio). Inocula were acclimated to these same urine-to-feces ratios to determine methane production. Results show that both urine-free and as-excreted substrates were not inhibitory to anaerobic inocula. Anaerobic microorganisms can be readily acclimated to substrate with double the as-excreted urine concentration, which contained nitrogen concentrations up to 7.20 g/L. Cumulative methane production reached similar levels in the batch tests, regardless of the substrate urine concentration. PMID- 18419014 TI - Modification of a full-scale sequencing batch reactor operational mode for biological nutrient removal. AB - Two biological nutrient removal modes, consisting of anaerobic, anoxic, and oxic sequences, were tested in a full-scale sequencing batch reactor. The modes, identified as BNR-S1 and BNR-S2, had average total nitrogen removals of 84 and 89%, respectively, for the months of August to October. Over the same period, total phosphorus removals for BNR-S1 and BNR-S2 were 88 and 87%, respectively. In contrast, total nitrogen and total phosphorus removals for the regular aerobic mode were 54.7 and 44.7%, respectively. When the wastewater temperature changed from approximately 20 to 15 degrees C in the winter months, total nitrogen and total phosphorus removals for BNR-S2 were reduced to 81 and 70%, respectively. Total nitrogen effluent concentrations were between 2.5 and 4 mg-N/L (at approximately 20 degrees C), while the effluent total phosphorus concentrations were between 1 and 2 mg/L. The BNR-S2 mode was found to require less energy per kilogram of soluble chemical oxygen demand removed than the regular and BNR-S1 modes. PMID- 18419016 TI - Biodegradation of disodium cocoamphodiacetate by a wastewater microbial consortium. AB - Biodegradation of an amphoteric surfactant commonly used in personal care products, disodium cocoamphodiacetate (DSCADA), was evaluated. Results from respirometry experiments indicated that high levels of DSCADA (>216 mg/L) may be toxic to bacteria in wastewater treatment processes. Limited biodegradation, with 50% dissolved organic carbon (DOC) removal and 80% chemical oxygen demand removal was observed in batch assays, while complete removal of the parent compound, DSCADA, was noted. Oxygen biosensors were used to evaluate biodegradability of the metabolites present in the batch samples. Additional aerobic microbial activity was not detected in these samples, even with a residual DOC of approximately 45 mg/L. Results from this research indicate that biodegradability of DSCADA is limited and recalcitrant metabolites may be formed. Because DSCADA is a commonly used surfactant and is present in domestic and industrial wastewater, the associated risk posed by residual compounds should be carefully evaluated. PMID- 18419017 TI - Concentration and recovery of coliphages from water with bituminous coal. AB - Coliphages represent a process indicator for fecal pollution. The coal bed concentration method prepared for enterovirus was refined for a concentration of coliphages. A bed made from 1.5 g of 120-mesh coal powder was used for concentrating coliphage from 200 mL of a water sample with or without the addition of aluminum chloride at different pH values. The isolated E. coli strain EC-R8 was found to be more susceptible to the desired coliphage and showed significant coliphage-coliform response, with clear plaque used for further studies. The complete coliphage adsorption was achieved with the addition of 0.0005 M AlCl3 at pH 6.0. Adsorbed coliphages were eluted with 3% beef extract in Mcllvaine buffer at pH 7.1, with an average recovery of 78.74%. This concentration technique was applied for the detection of coliphages from the well water of Nagpur city (India) and found to contain coliphages in the range of 2 to 28 plaque-forming units per liter (PFU/L). PMID- 18419018 TI - What procedural skills do deployed US navy family physicians need? PMID- 18419019 TI - Tactical combat casualty care. PMID- 18419020 TI - Department of Defense Global Emergings Infections Surveillance and Response System. PMID- 18419021 TI - Opportunity for optimization and integration: a view from the field. PMID- 18419022 TI - Health-related impact of deployment extensions on spouses of active duty army personnel. AB - This study examined problems pertaining to the health and well-being of Army spouses during deployment, comparing those whose experienced extensions of their partners' deployments with those whose partners returned home on time or early. It used data from a 2004 survey of 798 spouses of active duty personnel. Controlling for demographic and deployment characteristics, spouses who experienced extensions fared worse on an array of measures, including mental well being (e.g., feelings of depression), household strains (e.g., problems with household and car maintenance), and some areas of their jobs (having to stop work or to work fewer hours). There were no statistically significant differences regarding problems pertaining to their overall health, marriage, other work issues, finances, relationships with Army families, or safety. However, spouses who experienced extensions were more likely to perceive the Army negatively during deployment. These findings suggest that deployment extensions may exacerbate certain problems and frustrations for Army spouses. PMID- 18419023 TI - Sleep disturbance during military deployment. AB - This preliminary investigation evaluated symptoms of sleep disturbance and insomnia in a group of 156 deployed military personnel. A 21-item Military Deployment Survey of Sleep was administered to provide self-reported estimates of a variety of sleep parameters. The results indicated that 74% of participants rated their quality of sleep as significantly worse in the deployed environment, 40% had a sleep efficiency of < 85%, and 42% had a sleep onset latency of > 30 minutes. Night-shift workers had significantly worse sleep efficiency and more problems getting to sleep and staying asleep as compared to day-shift workers. The results of the study indicate the need for programs to help deployed military members get more and better sleep. PMID- 18419024 TI - Operation Aftershock: the U.S. military disaster response to the Yogyakarta earthquake May through June 2006. AB - The U.S. military has recently been involved in many humanitarian assistance and disaster response missions around the world. This newfound role is in response to the U.S. government's desire to use "medical diplomacy" rather than "military might" to shape its relationship with foreign governments. With each of these humanitarian assistance and disaster response missions, the U.S. military has learned how to more rapidly insert desperately needed services and skill sets into disaster-struck communities, how to arrange for in-country services (translation services, transportation, etc.) that cannot be readily brought in, and how to work closely with foreign governments and nongovernmental organizations to determine their needs and expectations without the U.S. military appearing as if it were trying to establish a permanent presence. PMID- 18419025 TI - Intensive coping skills training to reduce anxiety and depression for forward deployed troops. AB - This study provides descriptive information and 2-year outcome data on the first intensive, outpatient, coping skills training program for forward-deployed troops in the Western Pacific Ocean region. Established in February 2003 by the Mental Health Department of the U.S. Naval Hospital in Okinawa, Japan, the Outpatient Crisis Prevention Program was designed as a multidisciplinary training program to reduce anxiety and depression and to enhance the coping skills of active duty personnel. This study describes the rationale for creating the program, the patient population, and treatment outcome data collected during the first 2 years. The Beck Depression Inventory II and the Beck Anxiety Inventory were administered as pretraining and posttraining measures to patients who completed the program between February 2003 and February 2005 (N = 326). Results revealed that the Outpatient Crisis Prevention Program was effective in reducing depression and anxiety symptoms and promoting healthy coping behaviors among participants. Follow-up data indicated that gains were maintained 1 month after treatment. PMID- 18419026 TI - Injury and illness casualty distributions among U.S. Army and Marine Corps personnel during Operation Iraqi Freedom. AB - The objective of this study was to evaluate the distributions of U.S. Marine Corps and Army wounded in action (WIA) and disease and nonbattle injury (DNBI) casualties during Operation Iraqi Freedom Major Combat Phase (OIF-1) and Support and Stability Phase (OIF-2). A retrospective review of hospitalization data was conducted. chi2 tests were used to assess the Primary International Classification of Diseases, 9th Revision (ICD-9), diagnostic category distributions by phase of operation, casualty type, and gender. Of the 13,071 casualties identified for analysis, 3,263 were WIA and 9,808 were DNBI. Overall, the proportion of WIA was higher during OIF-1 (36.6%) than OIF-2 (23.6%). Marines had a higher proportion of WIA and nonbattle injuries than soldiers. Although overall DNBI distributions for men and women were statistically different, their distributions of types of nonbattle injuries were similar. Identifying differences in injury and illness distributions by characteristics of the casualty population is necessary for military medical readiness planning. PMID- 18419027 TI - Contract case managers prove cost effective in federal workers' compensation programs. AB - OBJECTIVE: This pilot study examined whether contract case managers are cost effective in reducing workers' compensation (WC) costs and preventing injuries. METHODS: We placed contract medical case managers at four installations where they employed private industry best practices. A needs assessment was performed and site-specific targeted interventions were developed. Case managers improved WC program effectiveness by ensuring command support, and strengthening the case management process. They minimized lost work time and provided the WC team resources to review the oldest cases. RESULTS: Case managers cost $1.25 million and they generated $4.4 million in savings for a 3.52 return on investment. Case managers removed 73 workers from long-term rolls by making use of vocational rehabilitation, finding modified duty jobs, offering workers medical retirement, and terminating benefits when workers refused to work after being offered a job. CONCLUSIONS: This study of medical case managers demonstrates they can reduce WC costs. PMID- 18419028 TI - Screening tests detect knee pain and predict discharge from military service. AB - This study evaluated the capacity of a screening test to predict discharge from military training. When starting their training, 590 artillery and 258 ranger conscripts were tested in step-up, step-down, rising from a 0.40-m-high bench, and bilateral squat tests, with pain intensity ratings. Ranger conscripts who did not complete their training were noted. The rising test identified 82% and the step-down test 81% of artillery conscripts who reported pain at any level during any of the screening tests. Receiver operating characteristic curve analysis revealed that both the step-down test and the rising test were good in predicting discharge from military ranger training because of knee problems. The step-down test identified 80% of ranger conscripts who did not complete training because of knee disorders. The tests constitute a simple, time-saving, cost-effective tool in a systematic process for screening knee pain to identify high-risk groups, for prioritization of interventions. PMID- 18419029 TI - Evaluation of body fat estimated from body mass index and impedance in Belgian male military candidates: comparing two methods for estimating body composition. AB - OBJECTIVE: The study objective was to evaluate whether a classification based on body mass index (BMI) agrees with a classification based on body fat mass, estimated by bioelectrical impedance. METHODS: A random sample of 448 male candidates between 18 and 20 years was selected during their medical visit in a military recruitment center. BMI was determined as weight/height2 and was considered normal between 20.0 and 25.0 kg/m2 (cfr. WHO classification). Percentage of body fat was estimated with bioelectrical impedance, using the Omron Body Fat Analyzer HBF-306. Subjects with a body fat percentage measured by bipolar bioelectrical impedance analysis (BF%(IMP)) < or = 20.9% were considered normal weight, while subjects with a BF%(IMP) > or = 21.0% were considered overweight. We used the following classification: true positives were normal scores for BMI and impedance; false positives were normal scores for BMI but not for impedance; true negatives were overweight scores for BMI and for impedance; and false negatives were overweight scores for BMI but not for impedance. Data were analyzed using the SPSS statistical program. RESULTS: BMI ranged from 17.0 to 29.4 kg/m2; percentage of fat mass varied between 5.3 and 31.4% of body weight. Of the total sample, 328 (73.2%) candidates were classified as true positive, 29 (6.5%) as false negative, 47 (10.5%) as false positive, and finally 44 (9.8%) as true negative. The difference in classification in normal weight versus overweight between the BMI method and the bipolar bioelectrical impedance method was statistically significant (chi2 with one df = 86.04; p < 0.001). CONCLUSIONS: To limit false-negative classifications, additional impedance measurements in the BMI category between 25.0 and 27.0 kg/m2 is mandatory to determine whether there is really an excess fat mass. PMID- 18419030 TI - Reported stress and its relationship to tobacco use among U.S. military personnel. AB - In addition to common stressors, members of the U.S. Armed Forces experience a high level of stress unique to their status as service members. In an effort to combat stress, many military personnel report high levels of nicotine use. This study investigated the relationship between tobacco use and perceived stress among military members in all four armed services. Results indicate that those who use tobacco products specifically to reduce stress report significantly higher stress levels than those who do not use tobacco. Moreover, current users and those who both smoked and used smokeless tobacco were far more likely to report experiencing "a lot" of stress from a variety of sources than former or never users. Tobacco users also engaged more frequently in negative coping behaviors and relied less on the positive coping strategies used more often by former or never smokers. These findings are consistent with the larger body of literature suggesting that tobacco is not only an ineffective stress-reducing strategy, it also likely perpetuates a stress response in users. It is, therefore, critical that the military improve effective means of coping among nicotine-using troops. PMID- 18419031 TI - Risk factors for training injuries among British Army recruits. AB - This study aimed to identify risk factors for training injuries resulting in referral to a remedial instructor (RI) or medical discharge (MD) among British Army recruits undertaking initial training. Physical performance and anthropometric data for 11,937 male and 1,480 female recruits were examined as potential risk factors for RI referral and MD, using Cox regression. There was a trend showing that female recruits' MD rates were higher than male recruits' rates (p = 0.096), and RI referral rates were significantly greater for women than for men (p = 0.041). The independent risk factors for MD were 2.4-km run time, ethnicity, and Army training regiment attended (p < 0.001), and those for RI referral were 2.4-km run time, ethnicity, Army training regiment attended, and body mass index (p < 0.001). Gender was not an independent risk factor for injury, suggesting that lower levels of aerobic fitness are the primary cause of the greater incidence of injury among female recruits during British Army initial training. PMID- 18419032 TI - Medical support for Pershing's Punitive Expedition in Mexico, 1916-1917. AB - Pershing's Punitive Expedition had adequate medical support despite deliberately limited in-theater resources. The few casualties did not strain the inadequate number of forward providers. Preventive medicine was highly successful due to significant medical and command emphasis. New technologies were useful and helped minimize the medical footprint. National Guard troops mobilized to support the Expedition had troublesome medical readiness rates. PMID- 18419033 TI - Diagnostic pelvic computed tomography in the rectal-injured combat casualty. AB - BACKGROUND: The current standard for evaluating trauma patients for penetrating rectal injury is to perform a rigid proctoscopy. This can be laborious and inaccurate. Injuries are often not visualized and a small number of unnecessary colostomies may be created. Computed tomography (CT) scanning of the pelvis may be useful in identifying penetrating rectal injuries. STUDY DESIGN: A retrospective analysis was performed on data regarding all casualties admitted to the 10th Combat Support Hospital during the period of November 2005 through March 2006. Nineteen patients were identified. Patients that were hemodynamically stable underwent preoperative CT scanning. All rectal injuries diagnosed preoperatively were confirmed through a different diagnostic modality in the OR. RESULTS: Nineteen patients with rectal injury or suspected rectal injury were identified. Eight of the 19 were hemodynamically unstable in the emergency medical treatment area and were taken emergently to surgery. For discussion, only stable patients with gunshot wound or blast/fragmentation injury mechanisms were included. No injuries were missed by CT scanning, but there were two false positive scans. CONCLUSIONS: In our brief experience, CT scanning was a useful screening tool to assist in identifying patients with penetrating traumatic rectal injuries. It allowed us to improve triage and make effective use of limited operative resources. PMID- 18419034 TI - Don't know, don't care: revisited. AB - OBJECTIVES: This study re-evaluates the awareness and attitudes of medical students concerning military programs, which can provide financial support for medical student education. An original study, 10 years ago, demonstrated a minimal awareness of these programs. With medical school tuitions rising dramatically in the intervening years, as well as the resultant student indebtedness increasing significantly, it is appropriate to readdress this knowledge of attitude toward financial options through military programs for medical students. Reassessment was undertaken using the same evaluation instrument. In the decade in which these four assessments were made, attitudes toward amelioration of medical student tuitional indebtedness, through programs sponsored by the Department of Defense, have worsened. This phenomenon jeopardizes military medicine and, in so doing, negatively impacts readiness of our Armed Forces. PMID- 18419035 TI - Emergency contraception knowledge, attitudes, practices, and barriers among providers at a military treatment facility. AB - The safety and efficacy of postcoital administration of oral contraceptives, commonly called "emergency contraception pills" (ECPs), have been well documented; nonetheless, this method of preventing pregnancy is underused in the United States. Through the use of surveys, the current study assessed providers' knowledge, prescribing practices, attitudes, and barriers regarding ECPs at a military treatment facility in the southwest United States. Results showed a lack of knowledge of ECP provision, with 34% believing treatment had to be initiated in 48 or fewer hours, and 60% believing two doses must be taken within 24 hours or more of each other. Slightly more than one- half (54.4%) of the providers reported having prescribed an ECP; those who had prescribed had more positive attitudes about ECPs. Perceived barriers included women not asking for ECPs, concerns about safety of ECPs, and issues regarding liability. A discrepancy between providers' perceived and actual knowledge about ECPs supports the need for better education for all providers who care for women. PMID- 18419036 TI - Completeness and timeliness of Chlamydia trachomatis and Neisseria gonorrhoeae genital infection reporting in the U.S. Air Force. AB - Genital Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (GC) are reportable diseases in the U.S. Air Force. Base-level personnel enter laboratory-confirmed cases into the Air Force Reportable Events Surveillance System (AFRESS). To examine completeness and timeliness of CT and GC reporting, we compared laboratory and AFRESS records collected between 2000 and 2005. We used joinpoint regression analysis to identify trends in reporting over time. Of 23,118 CT laboratory-confirmed cases, 74.0% were reported into AFRESS; of 1,897 GC cases, 57.5% were reported. CT reporting completeness increased through 2003, but declined afterward; completeness of GC reporting steadily declined throughout the study. The median time between collection and reporting dates was 12.0 days and 13.5 days for CT and GC, respectively. Timeliness of CT reporting improved until mid-2004, after which it sharply declined; GC reporting timeliness declined throughout the study. Monitoring data, training personnel, and adopting an electronic reporting system may improve data quality. PMID- 18419037 TI - Bondsteel fixed partial denture: a provisional indirect technique for in-the field dental emergencies. AB - This article presents a modified resin-bonded fixed partial denture technique of replacing a single missing anterior tooth under limitation of material and time. This technique involved a small section of an oral surgery arch bar that was bent and fitted passively into the grooves that were prepared in the lingual of the abutment teeth. A polycarbonate crown was fitted passively on the arch bar by a thin layer of Vaseline. The polycarbonate crown was then filled with acrylic resin to bond the polycarbonate crown and the arch bar together. The result provides an esthetic appearance and a 4-month clinical success without complication. Thus, this technique is a short-term solution for clinics that have limited resources for a definitive treatment. PMID- 18419038 TI - Helicopter mishap attributed to single seizure. AB - A case report is presented of a 36-year-old U.S. Coast Guard aviator who had a single seizure while operating a helicopter on the ground. His seizure activity produced a loss of consciousness during which he pushed the cyclic to the left anterior quadrant that resulted in a ground mishap. No risk factors were identified in an extensive neurological workup. The current guidance for handling seizures in military aviation personnel is reviewed, along with considerations for treatment. Although the military aviation selection process carefully screens applicants for seizure history and potential, occasional seizures in the aviation population remain possible. Such events may result in military aircraft mishaps despite careful risk factor surveillance, as demonstrated by this case. PMID- 18419039 TI - Evaluation of anomalous systemic and pulmonary venous return in a young child with heterotaxy: a case study--implications for military dependent care. AB - The case of a young male patient with a complex admixture lesion who required a comprehensive anatomic evaluation before palliative cardiac surgery is presented. We describe a safe anesthesia protocol for obtaining the late-acquisition, gadolinium-enhanced, magnetic resonance angiographic images necessary to define the complex pulmonary and systemic venous anatomic features of his cardiac admixture lesion. Subspecialty physician staffing implications for the care of military dependants with congenital heart disease who might benefit from evaluation using this safe simple protocol and readily available magnetic resonance imaging technology are addressed. PMID- 18419040 TI - Ischemic colitis of the ascending colon: a diagnostic and management conundrum. AB - Ischemic colitis is the most common form of intestinal ischemia. It most commonly involves the left side of the colon presenting with acute onset of abdominal pain followed by bloody diarrhea. Involvement of only the right or ascending colon is an infrequent occurrence. Because this problem is less recognized than its counterpart involving the left colon, the correct diagnosis and management may not be readily considered. We present a case of ischemic colitis presenting as a distal small bowel obstruction with emphasis on evaluation and management of this unusual clinical problem. PMID- 18419041 TI - Case #21: Superior Vena Cava (SVC) syndrome. PMID- 18419042 TI - Reconceptualizing the informed consent process at eight innovative hospitals. AB - BACKGROUND: Hospitals struggle to make informed consent processes understandable for patients. Amid "promising practices" for patient-centered communication, there is debate over how to handle consent processes for patients with limited literacy, health literacy, or English proficiency. METHODS: In 2005, the American Medical Association's Ethical Force Program and the American Hospital Association's Health Research and Educational Trust conducted eight site visits to determine how hospitals use patient-centered communication to improve health care. FINDINGS: Each of the eight hospitals noted a number of problems in the traditional informed consent process, sparking a broad set of efforts to improve. Disagreements about how to do so often focused on whether to attempt simplification and translation of informed consent documents. Specifically, hospital leaders held widely divergent views on legal and accreditation requirements for these forms. However, several other promising practices met with widespread approval. DISCUSSION: Overall, informed consent in the eight hospitals was becoming more patient centered and integrated. The significant variation across states makes it difficult to provide overarching policy guidance on the informed consent process. Greater regulatory clarity is needed so that policies and practices can be more closely aligned with the ethical foundations of informed consent. PMID- 18419043 TI - The cost consequences of improving diabetes care: the community health center experience. AB - BACKGROUND: Despite significant interest in the business case for quality improvement (QI), there are few evaluations of the impact of QI programs on outpatient organizations. The financial impact of the Health Disparities Collaboratives (HDC), a national QI program conducted in community health centers (HCs), was examined. METHODS: Chief executive officers (CEOs) from health centers in two U.S. regions that participated in the Diabetes HDC (N = 74) were surveyed. In case studies of five selected centers, program costs/revenues, clinical costs/revenues, overall center financial health, and indirect costs/benefits were assessed. RESULTS: CEOs were divided on the HDC's overall effect on finances (38%, worsened; 48%, no change; 14%, improved). Case studies showed that the HDC represented a new administrative cost ($6-$22/patient, year 1) without a regular revenue source. In centers with billing data, the balance of diabetes-related clinical costs/revenues and payor mix did not clearly worsen or improve with the program's start. The most commonly mentioned indirect benefits were improved chronic illness care and enhanced staff morale. DISCUSSION: CEO perceptions of the overall financial impact of the HDC vary widely; the case studies illustrate the numerous factors that may influence these perceptions. Whether the identified balance of costs and benefits is generalizable or sustainable will have to be addressed to optimally design financial reimbursement and incentives. PMID- 18419044 TI - Using patient traffic control to reduce treatment delays for high-risk patients at a VA hospital. AB - BACKGROUND: For patients at high risk of function-limiting or life-limiting disease, the time elapsed between first clinical presentation, diagnosis, and treatment can influence the likelihood of treatment success. METHODS: A systematic change in the management of high-risk patients was undertaken. This approach includes identifying primary provider responsibility, establishing communication expectations between providers, developing a tracking system to actively monitor patients (patient traffic control), and using a time guideline to assess patient progression. A 60-day time frame was established for the time from first clinical presentation to diagnostic exclusion or treatment initiation. RESULTS: In a one-year period, 288 high-risk patients were entered into patient traffic control, 211 (73%) of whom were monitored in the primary care setting. The median time to diagnostic exclusion or treatment was 43 days (mean, 58.5 days). Sixty-six percent of all patients achieved diagnostic exclusion or treatment by 60 days. Of the 95 patients monitored for > 60 days, 56% had delays caused by patient noncompliance or because of the appropriate need for long-term serial radiographic monitoring. Thirty-eight patients (13.1%) demonstrated problems with appointment nonadherence. None were lost to follow-up. DISCUSSION: The patient traffic control approach enabled the management of the majority of high-risk patients within 60 days of presentation. PMID- 18419045 TI - How often are potential patient safety events present on admission? AB - BACKGROUND: Data fields that capture whether diagnoses are present on admission (POA)--distinguishing comorbidities from potential in-hospital complications- became part of the Uniform Bill for hospital claims in 2007. The AHRQ Patient Safety Indicators (PSIs) were initially developed as measures of potential patient safety problems that use routine administrative data without POA information. The impact of adding POA information to PSIs was examined. METHODS: Data were used from California (CA) and New York (NY) Healthcare Cost and Utilization Project (HCUP) state inpatient databases for 2003, which include POA codes. Analysis was limited to 13 of 20 PSIs for which POA information was relevant, such as complications of anesthesia, accidental puncture, and sepsis. RESULTS: In New York, 17% of cases revealed suspect POA coding, compared with 1% 2% in California. After suspect records were excluded, 92%-93% of secondary diagnoses in both CA and NY were POA. After incorporating POA information, most cases of decubitus ulcer (86%-89%), postoperative hip fracture (74%-79%), and postoperative pulmonary embolism/deep vein thrombosis (54%-58%) were no longer considered in-hospital patient safety events. DISCUSSION: Three of 13 PSIs appear not to be valid measures of in-hospital patient safety events, but the remaining 10 appear to be potentially useful measures even in the absence of POA codes. PMID- 18419046 TI - Reducing accidental extubation in neonates. AB - Unplanned or accidental extubations (UPEs) showed little improvement until the clamp method of fixation of endotracheal tubes was selected. PMID- 18419047 TI - Improving the documentation of vital signs: a business reengineering efficiency study. AB - By upgrading its electronic medical record to include a Quick Vitals screen, the Henry Ford Health System increased the mean rate of entry of vital signs from 0.7% to 58.5%. PMID- 18419048 TI - Improving sepsis care through systems change: the impact of a medical emergency team. AB - The Hospital of the University of Pennsylvania achieved a significant reduction in the time between prescription and administration of antibiotics by embedding a pharmacist in an MET to facilitate antibiotic delivery. PMID- 18419049 TI - Immune intervention with monoclonal antibodies targeting CD152 (CTLA-4) for autoimmune and malignant diseases. AB - CD152 or cytotoxic T lymphocyte antigen-4 (CTLA-4) is an essential receptor involved in the negative regulation of T cell activation. Because of its profound inhibitory role, CD152 has been considered a sound susceptible candidate in autoimmunity and a persuasive target for cancer immunotherapy for over a decade. However, the precise roles played by this molecule continue to emerge. In particular, recent evidence suggests that CD152 is also important in the homeostasis and function of a population of suppressive cells, termed regulatory T cells (Treg). In this review, we discuss the recent progress and main features of monoclonal antibodies (mAbs) targeting CD152 and examine how each mAb prepared to a distinct epitope may impact differently upon CD152 modulation depending on its demonstrated regulatory role acting as an agonist, antagonist, or inverse agonist. PMID- 18419050 TI - Hepatitis C virus genotypes: clinical relevance and therapeutic implications. AB - The incidence of hepatitis C virus (HCV) -related hepatocellular carcinoma (HCC) has been increasing in several countries including Taiwan. There are six main genotypes, each of which contains closely related subtypes. Molecular epidemiological studies have shown marked differences in the genotype distribution by geographical region and between patient groups. HCV genotype 1 may play a role in the development of HCC, although some studies have argued against this. A sustained virological response secondary to interferon monotherapy or interferon/ribavirin combination therapy may reduce the risk of HCC and improve survival in chronic hepatitis C patients. The HCV genotypes are associated with therapeutic response. Rapid virological response is also a predictor of therapeutic response. Although viral characteristics have consistently been shown to be important predictors of treatment response, identification of additional host immune and genetic factors involved in determining the outcome of antiviral therapy is necessary. Newly developed bio techniques (microarray, proteomes, bioinformatics), drugs, and treatment strategies may elucidate the pathogenesis and improve the therapeutic response in HCV infection. PMID- 18419051 TI - Defining the carbohydrate specificities of Erythrina corallodendron lectin (ECorL) as polyvalent Galbeta1-4GlcNAc (II) > monomeric II > monomeric Gal and GalNAc. AB - BACKGROUND: Erythrina corallodendron lectin (ECorL) is one of the potent applied lectins. In previous studies, the carbohydrate specificities of this lectin were limited to monosaccharides, simple oligosaccharides and several clusters. However, the polyvalent factor has not been investigated. METHODS: The binding properties at the combining sites of ECorL were characterized by sensitive enzyme linked lectinosorbent (ELLSA) and inhibition assays, using our collection of ligands and polyvalent natural glycans with known glycotopes. RESULTS: Results of both binding and inhibition assays revealed a very high affinity between ECorL and Galbeta1-4GlcNAc (II)-containing glycoproteins. Among soluble natural glycans tested for inhibition, the high-density polyvalent II glycotopes, such as Streptococcus pneumoniae type 14 capsular polysaccharide which is composed of repeating poly-II residues, resulted in 2.4 x 10(4), 1.4 x 10(3) and 8.6 x 10(2) fold higher affinities to ECorL than the monomeric Gal, linear II and tri antennary II, respectively, at the non-reducing end in N-linked glycopeptides (Tri-II). The ECorL-glycan interaction was also strongly inhibited by most of the other high-density II-containing glycoproteins. Although GalNAc was as potent an inhibitor as Gal, its polyvalent structural units were poor inhibitors. CONCLUSIONS: [1] Galbeta1-4GlcNAc (II) and other Galbeta1 -related oligosaccharides are essential for binding. [2] Their polyvalent form in glycoproteins is the most important binding factor for ECorL, while II monomer and oligo-antennary II forms play only a limited role in binding. [3] Although GalNAc is more active than Gal for ECorL, its reactivity is not changed by polyvalent effects. This lectin may be used as a tool to study glycobiology in basic and medical sciences. PMID- 18419052 TI - Effects of retinoic acid on morula-stage embryo development in mice. AB - BACKGROUND: In a previous study, we investigated the short-term effects of in utero retinoic acid (RA) exposure on early development at the cleavage stage before implantation to understand the possible roles of RA in blastocyst formation. We designed the present study to investigate the long-term effects of RA exposure on pre-implantation embryos in vivo and in vitro. METHOD: To evaluate late pre-implantation exposure to RA, pregnant female mice were fed peanut oil with 50 mg/kg RA by oral gavage in the late afternoon of day 2 and early morning of day 3 of gestation. Mice were sacrificed on day 8. The number and morphology of embryos were recorded. The morula-stage embryos were treated with different doses of t-RA for 1 day and were cultured for the following 7 days in vitro. The post-implantation development in vitro was evaluated. RESULTS: The in vivo study showed that the rate of blastocyst implantation was not significantly different (9.2 vs 10.2 per mouse) and the rate of post-implantation embryo resorption was significantly higher in retinoic acid treated mice than in controls (35% vs 0%). RA was administered to morula-stage embryos at levels of 0, 0.001 micro, 0.1 micro, or 10 micro M throughout in vitro culture. For embryos that continued to develop following implantation, the development stages were delayed when assessed in 7-day culture. The percentage of embryos in the later stages of development changed depending on the dose of retinoic acid used. CONCLUSION: These findings document, for the first time, that RA exerts adverse effects on morula-stage embryos during the early stage of development. PMID- 18419053 TI - Primary intraocular lens implantation for unilateral idiopathic cataract in children. AB - BACKGROUND: To examine the efficacy of intraocular lens implantation on visual rehabilitation and ocular growth in children with unilateral idiopathic cataracts. METHODS: Preoperative characteristics, visual outcome and postoperative refraction change were analyzed through a retrospective study of the charts of children who underwent cataract removal and primary intraocular lens implantation for unilateral idiopathic cataracts between 1994 and 2003. RESULTS: Thirty-one children were included in this study. Preoperatively, 83% of the eyes with cataracts had best corrected visual acuity (BCVA) of less than 20/200. Posterior subcapsular opacity was the most common type of cataract. The average age at surgery was 66.8 months (range 14-115 months) and the mean follow up was 43.4 months (range 12-117 months). At the final visit, there was no significant difference in refractive development between the pseudophakic eyes and the fellow eyes of the subjects. Sixty percent of the pseudophakic eyes had a BCVA of 20/200 or better and 37% had a BCVA of 20/60 or better. Preoperative visual acuity was the only significant factor related to visual outcome. CONCLUSION: Primary intraocular lens implantation in children with unilateral idiopathic cataracts is an effective treatment for visual rehabilitation. The refractive development of the pseudophakic eyes was not significantly different from the fellow eyes. PMID- 18419054 TI - Pediatric ocular trauma in Taiwan. AB - BACKGROUND: Ocular trauma is a significant problem in pediatric patients. The aim of this investigation was to evaluate the demographic, etiologic and prophylactic factors of pediatric ocular trauma in a tertiary center in northern Taiwan. METHODS: A retrospective review of medical charts of patients aged 15 years or younger who visited the emergency department of Chang Gung Memorial Hospital, Taoyuan for ocular trauma between July 2003 and June 2004 was undertaken. RESULTS: Of the 228 children in this study, ocular trauma occurred more frequently in boys than in girls (151:77 patients, p < 0.0001). Twenty-one patients (9.2%) presented with bilateral trauma. The most common ocular injury was closed globe injury (178 patients, 78.1%). The next most common ocular injuries were burns (35 patients, 15.4%), open globe injuries (8 patients, 3.5%) and orbital injuries (7 patients, 3.1%). Falling was the most common cause of ocular trauma in these children, followed by trauma caused by a fist or other part of the body. Sharp objects such as knives or scissors were the most common cause of open globe injuries. Visual impairment (visual acuity less than 20/50) occurred in three open globe injury patients. CONCLUSIONS: Ocular trauma in children is associated with visual loss. Many of the cases were preventable. Public education, general awareness and aggressive primary management may be indicated to optimize visual outcome. PMID- 18419055 TI - Psychiatric manifestations in young females with congenital adrenal hyperplasia in Taiwan. AB - BACKGROUND: Congenital adrenal hyperplasia (CAH) is a syndrome of prenatal and/or postnatal androgen excess secondary to genetic deficits in the cytochrome p450 enzymes of the cortisol synthesis pathway. Women with CAH may suffer from different degrees of androgenization. This study documented psychiatric manifestations in young women born with CAH in Taiwan. METHODS: From July 1, 2005 to February 31, 2006 the psychiatric morbidity, gender identity and sex-typed behavior during childhood of 11 young women with CAH were assessed either using questionnaire-based semi-structured interviews or self-reported questionnaires. RESULTS: Of the 11 young women (mean age 15.3 +/- 5.6 years), 3 (27.3%) had salt wasting CAH and 8 (72.7%) had simple-virilizing CAH. Two patients without prior gender assignment (delayed diagnosis) were evaluated for primary amenorrhea at age 19 and 24 years, respectively. Four of the eleven participants (36.4%) had had an axis I psychiatric diagnosis within the past year. The subjects with more atypical sex-typed behavior in childhood tended to have a higher risk of minor psychiatric illness (r = -0.706, p < 0.05). CONCLUSIONS: This study examined the impact of CAH on the patients' psychological wellbeing. Psychiatric needs were found to be unmet. Earlier psychiatric evaluation and intervention for these patients is suggested. PMID- 18419056 TI - Hepatosplenic fungal infection in adult patients with acute leukemia. AB - BACKGROUND: Hepatosplenic fungal infection is an important infectious complication in adult patients with acute leukemia. METHODS: From 2001 to 2004, 163 adult patients were diagnosed with acute leukemia at our center: 41 patients had acute lymphoblastic leukemia (ALL) and 122 patients had acute myeloid leukemia (AML). Their charts were retrospectively reviewed. RESULTS: Of these 163 patients, 16 patients (9.8%) developed hepatosplenic fungal infection: three were ALL patients and 13 were AML patients. All of these patients suffered from febrile neutropenia after chemotherapy. Duration of agranulocytosis (absolute neutrophil count < 500/dl) was 10 to 36 days, with a median of 20 days. Clinical presentations in these patients were fever (94%), diarrhea (50%), abdominal pain (44%), oral mucositis (44%), papular skin lesions (31%) and lower back pain (7%). Fourteen patients (88%) had elevated alkaline phosphatase levels between 197 U/l to 1172 U/l (normal range: 28-94 U/l). The most common infection sites found by computed tomography were the spleen (94%) and the liver (88%). All patients were treated with antifungal agents. No patient died as a result of the fungal infection episode. Nine patients (56%) died due to uncontrolled underlying hematological malignancies. The median duration of follow-up was 15.2 months (range: 2.3-47.4 months). CONCLUSION: Alkaline phosphatase level and computed tomography are useful tools for the diagnosis of hepatosplenic fungal infection. Infection-related mortality is very low with effective treatment. Treatment for underlying diseases should proceed as soon as possible if the infection has been controlled. PMID- 18419057 TI - Implantable cardioverter defibrillator therapy: ten years experience in a medical center. AB - BACKGROUND: An implantable cardioverter defibrillator (ICD) is the therapy of choice for survivors of life-threatening ventricular tachyarrhythmias or sudden cardiac death. To date there is little data concerning the clinical features and outcome of ICD therapy among Taiwanese. This study identifies factors related to the outcome of ICD therapy over a ten-year period at this institution. METHODS: Forty-nine ICDs were implanted in 46 patients between August 1996 and January 2006. The mean follow-up duration was 32 +/- 21 months. Patient data, primary cardiac diagnosis, presenting cardiac arrhythmia, echocardiographic parameters, hemodynamic indexes, electrophysiologic findings, and follow-up observations were analyzed. The findings were compared to those of the Taiwan ICD Multicenter Registry (TIMR) Study and major secondary prevention ICD trials in the literature. RESULTS: The patients in this study were comparable to those of TIMR but were younger and had better left ventricular ejection fractions (LVEF) than those in Western countries. Furthermore, higher mortality on follow-up was observed in patients with any of the following: LVEF < 35%, New York Heart Association (NYHA) functional class III or IV, a left atrial dimension > or = 55 mm, a left ventricular end diastolic dimension > or = 75 mm, an end systolic dimension > or = 60 mm, triple vessel disease, a prior anterior myocardial infarction, and amiodarone or diuretic therapy. Patients with structural heart disease other than ischemic heart disease or dilated cardiomyopathy had higher event recurrence rates. CONCLUSION: Left ventricular function is a major determinant affecting the outcome in ICD recipients. Aggressive treatment for heart failure is warranted in these patients. PMID- 18419058 TI - Do closed-system hospitals shift care under case payment? Early experiences comparing five surgeries in Taiwan. AB - BACKGROUND: Recently, Japan, Korea, and Taiwan have adopted prospective payment systems (PPS) for healthcare. Experiences of the United States Medicare show that PPS reduces length of stay but creates incentives to shift care from regulated to un-regulated settings. In this study we investigated whether closed-system hospitals in Taiwan responded to case payment (CP) - one type of PPS, and if so, how this was managed. METHODS: Data were derived from three Taiwanese hospitals for five different surgical procedures (N = 22,327). The study period covered from October 1996 through August 1999, with CP commencing on October 1, 1997. Important dependent variables included inpatient medical claims, outpatient medical claims, and number of outpatient visits. Outpatient utilization from the period four weeks prior to admission and four weeks following discharge were merged for each patient. Ordinary Least Square (OLS) and Poisson regression were used to test the study's shifting hypotheses, controlling for gender, age, patient diagnoses, and institution attended. RESULTS: Length of hospital stay, amount of inpatient claims, and inpatient x-ray and lab-test claims were significantly reduced after CP. Corresponding OLS coefficients for the second year of implementation were, respectively, -.86, -.06, -.15, and -.04 (p < 0.01). Significant forward shifting of outpatient care, (79%), was found during the second year of CP. Despite the care-shifting effects noted herein, the average per-capita total claims reduced by 12%. Significant institutional effects were associated with the pattern of care-shifting. CONCLUSIONS: Our results indicate that CP reduced total claims for the selected surgical procedures, even under evident forward care-shifting. PMID- 18419059 TI - A successful live twin birth by in vitro fertilization after conservative treatment of recurrent endometrial cancer. AB - Endometrial cancer is predominately a postmenopausal disease. Endometrial cancer in women of childbearing age is relatively unusual. Endometrial cancer is typically treated with hysterectomy. After the development of endometrial cancer, successful pregnancy is rare. We present a case of recurrent stage I endometrial adenocarcinoma in a 35-year-old woman. Magnetic resonance imaging (MRI) revealed endometrial lesions without myometrium invasion and no pelvic lymph node enlargement. The patient refused surgical intervention with abdominal hysterectomy and bilateral salpingo-oophorectomy because of her essential desire for children. Fertility-preserving medical therapy with megestrol acetate for 1 year and subsequent assisted reproductive treatment (ART) were performed. Successful pregnancy occurred after in vitro fertilization-embryo transfer (IVF ET). On the basis of these observations and the low malignant potential of well differentiated endometrial carcinoma, fertility-preserving treatment using Megace therapy was suggested. In this case, recurrence occurred after the completion of Megace therapy and three failed attempts at artificial insemination by the husband (AIH). Recurrent endometrial adenocarcinoma was documented using hysteroscopy and direct endometrial biopsy. Another course of Megace therapy was administered due to her desire for children. A successful pregnancy occurred after long-term medical treatment and IVF-ET. PMID- 18419060 TI - A case of gastrointestinal stromal tumor with hyperinsulinemic hypoglycemia. AB - Gastrointestinal stromal tumor (GIST) is an uncommon malignant tumor that has recently been drawn to the attention of clinicians because of the protean clinical spectrum, endocrinological pathogenesis, aggressive metastatic features, poor prognosis, oncogenic tyrosine kinase receptor mutation and its brilliant revolutionized inhibitor imatinib. However, very few cases of GIST and hypoglycemia have been reported worldwide and they are usually associated with normal or low insulin levels. Here, we report on a rare case of GIST with postabsorptive hypoglycemia, extraordinary hyperinsulinemia and low insulinlike growth factor I (IGF-I). Its unusual neurological presentation made its diagnosis very difficult. After a complete resection, the symptoms diminished. We point out the unusual endogenous hyperinsulinism, clinical features and postulate possible mechanisms. PMID- 18419061 TI - Doctor of nursing practice (DNP). PMID- 18419062 TI - The clinical nurse leader: transforming care in the microsystem. PMID- 18419063 TI - [Genetic diversity of Hippophae rhamnoides L. sub-populations on loess plateau under effects of varying meteorologic conditions]. AB - The genetic diversity and genetic differentiation of eight Hippophae rhamnoides L. populations in eastern Gansu Province, as well as the effects of varying meteorologic conditions on them were assessed by using inter-simple sequence repeat (ISSR) molecular marker method. A total of 240 individuals in the populations were sampled. Using eleven primers, 165 bands were generated, ranging in size from 300 to 1500 bp, and 157 (95.76%) were found to be polymorphic. The analysis of molecular variance (AMOVA) demonstrated that there was a relatively high level (76.5%) of genetic variation within the populations, with the gene differentiation coefficient (Gst) and gene flow being 0.2418 and 1.5675, respectively. Therefore, to protect the gene resources of H. rhamnoides, the individuals within the populations should be first considered. Mantel test showed that genetic distance was significantly positively correlated with geographical distance (r = 0.65, p = 0.002), and regression modeling between genetic diversity and meteorologic factors suggested that there was a significant positive correlation between wind speed during blooming and genetic diversity of H. rhamnoides, illustrating that wind speed in blooming period and geographic distance were the vital factors affecting the genetic diversity of H. rhamnoides population. PMID- 18419064 TI - [Altitudinal changes in leaf mass per unit area and tissue non-structural carbohydrates content of Abies fabri on Gongga Mountain of southwest China]. AB - Taking the northeast slope of Moxi vally and southwest slope of Kangding vally in Gongga Mountain of Southwest China as sampling sites, this paper investigated the leaf mass per unit area (LMA) and tissue non-structural carbohydrates content (NSC) of Abies fabri distributed at the positions from lower elevations to alpine treeline during the periods of dormancy and vigorous growth. The results indicated that A. fabri had higher LMA and NSC in warm-moist Moxi valley than in hot-dry Kangding valley. The LMA and NSC of treeline trees were all higher than those of the trees at lower elevations, which was more obvious during the period of vigorous growth. The overall altitudinal changing trends of LMA and NSC in Gongga Mountain of Southwest China did not support the hypothesis of 'carbon limitation could induce the formation of alpine treeline'. PMID- 18419065 TI - [Effects of elevated CO2 concentration and temperature on nutrient accumulation and allocation in Betula albo-sinensis seedlings]. AB - With enclosed chamber system, this paper studied the effects of elevated CO2 concentration (ambient + 350 micromol x mol(-1), E(C)), temperature (ambient + 2 degrees C, E(T)), and their combination (E(CT)) on the nutrient accumulation and allocation in subalpine Betula albo-sinensis seedlings in western Sichuan Province. The results showed that after a growth season, the accumulation amount of N, P and K per plant increased by 44%, 45% and 11% under E(C) (P < 0.05), by 37%, 76% and 9% under E(T) (P < 0.05), and by 24%, 88% and 20% under E(CT) (P < 0.05), respectively. The N allocation to leaves reduced by 11.68% (P < 0.05) under E(C) but increased by 11.09% (P <0.05) under E(T) while that to branches, stems and roots increased by 2.95%, 3.39% and 5.34% under E(C) (P > 0.05), but decreased by 0.69%, 10.35% and 0.05% under E(T) (P > 0.05), respectively. The N allocation pattern under E(CT) was similar to that under E(C). The allocation of P and K had greater differences under E(C), E(T) and E(CT). All of these suggested that elevated CO2 concentration and temperature could promote nutrient accumulation, and change its allocation pattern in plant organs. PMID- 18419066 TI - [Ecological classification system of forest landscape in eastern mountainous region of Liaoning Province]. AB - Based on Digital Elevation Models (DEM) and satellite SPOT-5 data, and by using the spatial analysis function in Geographic Information System, a hierachical Ecological Classification System of forest landscape was developed for the eastern mountainous region of Liaoning Province, and the two lowest layers in the hierachical framework, Ecological Land Types (ELTs) and Ecological Land Type Phases (ELTPs), were mapped. The results indicated that there were 5 ELTs and 34 ELTPs. The boundaries of ELTs, which presented the potential vegetation distribution and potential forestry ecosystem productivity, were determined by environmental conditions quantified by DEM. ELTPs were classified by overlaying ELTs with forest vegetation data layers which were obtained from remotely sensed data, forest inventory data, and ground data. The ELTPs represented the divisions of land in terms of both natural and human-induced forest conditions, and therefore, were reliable units for forest inventories and management. ELTPs could function as conventional forest inventory sub-compartments. By this means, forestry departments could adjust forest management planning and forest management measures from the viewpoint of forest landscape scale to realize forest ecosystem management. PMID- 18419067 TI - [Invertebrate biodiversity in litter layers of natural forest and Eucalyptus plantation in eastern Guangdong, China]. AB - A comparative study was made on the community structure of invertebrates and the species diversity of thrips in the litter layers of natural forest and Eucalyptus urophylla plantation in eastern Guangdong of China. The results showed that in natural forest, Acarina, Collembola, Dipteran larvae, Hymenoptera, Thysanoptera, and Coleoptera were the most abundant invertebrates, accounting for 96.5% of the total individuals collected; while in Eucalyptus plantation, Acarina, Collembola, Dipteran larvae, and Lepidopteran larvae were the dominant invertebrate groups, which accounted for 96.3% of the total. The diversity of invertebrate assemblages was much higher in natural forest than in Eucalyptus plantation, based on the comparsions of Shannon-Wiener diversity index (H'), Pielou eveness index (J), Density-group index (DG), and Simpson dominance index (D). The individuals and species of fungus-feeding thrips were also more abundant in natural forest than in Eucalyptus plantation. However, there was no significant difference in the average density of invertebrates between natural forest and Eucalyptus plantation, because the individuals of Acarina were predominant, constituting 77.6% of the total. All of the results suggested that it is important to remain the understory and litter to improve the litter invertebrate diversity in fast growing Eucalyptus plantation. PMID- 18419068 TI - [Distribution characteristics and seasonal dynamics of Cu and Zn in shrub-marsh plants in mountainous areas of northeast China]. AB - The study on the distribution, accumulation, and seasonal dynamics of Cu and Zn in shrub-marsh plants Salix rosmarinifolia, Salix pentandra, Carex caespitosa and Carex schmidtii in mountainous areas of Northeast China showed that the Cu concentration in test plants varied from 6 to 12 mg x kg(-1), and its distribution was in the sequence of root > stem > leaf in S. rosmarinifolia and S. pentandra, and of stem > leaf > root in C. caespitosa and C. schmidtii, suggesting that Cu was mainly accumulated in the root of shrubs and the stem or leaf of Carex. Shrubs and Carex had less difference in their Cu concentration. The Zn concentration in test plants was 30-250 mg x kg(-1), and its distribution was in the sequence of leaf > stem > root in S. rosmarinifolia and S. pentandra, and of root > stem > leaf in C. caespitosa and C. schmidtii, indicating that Zn was mainly accumulated in the leaf of shrubs and the root of Carex. Shrubs had a higher Zn concentration than Carex. The accumulation coefficient of Zn in the organs of S. rosmarinifolia and S. pentandra was higher than 1.45, suggesting a good Zn-accumulation ability of these plants. The Cu and Zn concentrations in the aboveground parts of the four plants were higher during the initial growth period and then fluctuated to decrease with season, while those in roots were all higher both in the initial and in the late growth periods. PMID- 18419069 TI - [Seasonal fluctuation of soil microbial biomass carbon in secondary oak forest and Pinus taeda plantation in north subtropical area of China]. AB - With random block experimental design, the soil microbial biomass carbon, soil temperature, soil moisture, and litterfall input in secondary oak forest and Pinus taeda plantation were measured in successive two years at the Xiashu Experimental Forest of Nanjing Forestry University. The results showed that in the two stands, soil microbial biomass carbon had an obvious seasonal fluctuation, being lower in plant vigorous growth season but higher during plant dormancy. The microbial biomass carbon in 0-10 cm soil layer ranged from 267.8 mg x kg(-1) to 459.8 mg x kg(-1) in P. taeda plantation and from 278.6 mg x kg(-1) to 467.8 mg x kg(-1) in secondary oak forest. Soil microbial biomass carbon had a significant negative correlation with soil temperature, but no significant correlations with soil moisture and litterfall input. It was suggested that the seasonal fluctuation of soil microbial biomass carbon in test stands could be more related to the availability of soil carbon and other nutrients, competition of plant roots for soil nutrients, and plant growth rhythm. PMID- 18419070 TI - [Effects of water stress and temperature on gas exchange and chlorophyll fluorescence of Sinocalycanthus chinensis leaves]. AB - Sinocalycanthus chinensis is an endangered species in Sinocalycanthus, and only distributed in Zhejiang Province of China. This paper studied the photosynthetic responses of 2-year-old pot-cultured S. chinensis to different levels of water stress and temperature. The results indicated that under mild and moderate water stress, the net photosynthetic rate (Pn) of S. chinensis leaves was decreased to 92.3% and 74.3% of the control, respectively, which was mainly attributed to stomatal limitation; and under severe water stress, the Pn was decreased to 44.4% of the control, which might be mainly linked to non-stomatal limitation. The appropriate temperature for S. chinensis photosynthesis was from 20 degrees C to 28 degrees C. At 39 degrees C, the Pn, water use efficiency (WUE), and maximal photochemistry efficiency (Fv/Fm) were decreased significantly, while the dark respiration rate (Rd) and transpiration rate (Tr) were enhanced significantly. With increasing water stress and temperature, some photosynthetic parameters including light saturation point (LSP), apparent quantum yield (AQY) and maximal CO2 assimilation rate (Pmax) decreased to certain extents, while light compensation point (LCP) increased, suggesting that both severe water stress and higher temperature were the important environmental factors affecting the survival of S. chinensis. PMID- 18419071 TI - [Energy and nutrient characteristics of Carex lanceolata in Ziwuling of Loess Plateu]. AB - The study on the energy and nutrient characteristics of Carex lanceolata, a dominant companion species in different plant communities in Ziwuling forest area of Loess Plateau showed that the ash free caloric values of the aboveground and underground parts of the species tended to decline in the communities experienced different successional stages. The aboveground part of C. lanceolata had the highest ash free caloric value in Sophora viciifolia community and the lowest in Hippophae rhamnoides community. The underground part of C. lanceolata had the highest ash free caloric value in S. viciifolia community and the lowest in Quercus liaotungensis community. The aboveground part of C. lanceolata had a remarkably higher caloric value than its underground part, and the underground part appeared to extremely significantly differ in gross and ash-free caloric values among different communities. The caloric value of C. lanceolata varied greatly in the communities going through their early successional stages (S. viciifolia, H. rhamnoides, Populus davidiana and Betula platyphylla communities), and less greatly in the communities experiencing their later successional stages (Pinus tabulaeformis and Q. liaotungensis communities). There was a significant positive correlation between the gross caloric value and the C/N ratio in the underground part of C. lanceolata. PMID- 18419072 TI - [Effects of salt stress on cucumber seedlings root growth and polyamine metabolism]. AB - By the method of water culture, this paper studied the effects of NaCl stress on the seedlings root growth and polyamine metabolism of two cucumber cultivars 'Changchun mici' and 'Jin-chun No. 2'. The results showed that under NaCl stress, root growth was inhibited, and lipid peroxidation and electrolyte leakage increased. These changes were more obvious in salt-sensitive 'Jin-chun No. 2' than in salt-tolerant 'Changchun mici'. NaCl stress caused an increase of arginine decarboxylase (ADC), ornithine decarboxylase (ODC) and S adenosylmethionine decarboxylase (SAMDC) activities in cucumber roots, which was much greater in 'Changchun mici' than in 'Jinchun No. 2'. At the highest value, ADC, ODC and SAMDC activities increased by 149.3%, 60.1% and 69.4% in 'Changchun mici' and 118.6%, 56.2% and 50.6% in 'Jinchun No. 2', respectively, in comparison with the control. Diamine oxidase activity increased in 'Changchun mici' but not in 'Jinchun No. 2', and the increment of polyamine oxidase activity in 'Changchun mici' was smaller than that in 'Jinchun No. 2'. As a result, the contents of free spermidine and spermine and of conjugated and bound polyamines in 'Changchun mici' increased significantly, while that of free putrescine had a significant increase in 'Jinchun No. 2'. It could be concluded that higher levels of free spermidine and spermine and of conjugated and bound polyamines and lower level of free putrescine in roots could improve the adaptability of cucumber seedlings to salt stress. PMID- 18419073 TI - [Soil nutrient leaching patterns in maize field under different fertilizations: an in situ study]. AB - An in situ field experiment with lysimeter was conducted to study the effects of different fertilizations on the nutrient leaching loss from brown soil in the growth season of summer maize. The results showed that abundant rainfall and irrigation were the main factors affecting the leaching loss. The leaching amount was higher in the early growth period of summer maize, but decreased after then. The difference among different fertilization treatments also decreased with maize growth. Comparing with N fertilization, wheat stalk plus N application intensified the leaching. During the growth period of summer maize, the NO3- -N content in leached water in fertilization treatments had two peaks, while the NH4+ -N content had a trend of increased first and decreased then. The leaching loss of soil N was mainly in the form of NO3- -N. The accumulative leaching loss of NO3- -N was 12.90-46.53 kg * hm(-2), and that of NH4+ -N was 1.66-5.11 kg x hm(-2), both of which increased with increasing N application rate. The leaching rate of soil N was 6.53%-13.07% higher in treatment wheat stalk plus N application than in treatments of N fertilization, and 3.66%-10.10% higher in low N treatments than in high N treatments. The accumulative leaching loss of available P was only 0.148-0.235 kg x hm(-2), while that of available K was 7.08 13.00 kg x hm(-2). In the late growth period of summer maize, wheat stalk plus N application increased the leaching loss of soil available P and K, while nitrogen application affected it slightly. PMID- 18419074 TI - [Effects of long-term fertilization on phospholipid fatty acids and enzyme activities in paddy red soil]. AB - Soil samples were collected from the paddy fields at the Ecological Experimental Station of Red Soil, Chinese Academy of Sciences under different treatments of long-term fertilization, and their phospholipid fatty acids (PLFAs) and enzyme activities were determined. The results showed that soil enzyme activities, nutrient contents, microbial biomass, and PLFAs varied greatly with different fertilizations. Fertilization increased the kinds and amount of soil PLFAs. Compared with fertilized soil, unfertilized soil had more fungal PLFAs but less bacterial PLFAs, indicating that fungus was more adaptable to infertile soils than bacteria. Soils applied with NPK and organic fertilizer had higher amount of total PLFAs, which was 3.22 and 1.79 times higher than that under N fertilization and no fertilization. It was indicated that balanced fertilization with NPK or applying organic fertilizer was more beneficial to the growth of plants. Fertilization could also increase soil enzyme activities, and soil urease and phosphatase activities could be used as the indicators of soil fertility. PMID- 18419075 TI - [Adsorption characteristics of soluble organic carbon and nitrogen in two cultivated soils]. AB - In this paper, soluble organic carbon (SOC) and nitrogen (SON) were extracted from manure, and their adsorption characteristics in Argosols and Anthrosols in Guanzhong region of Shaanxi Province were investigated. The results showed that the adsorption of SON and SOC in the two soils could be fitted by initial mass isotherm model, and the adsorbed amounts of SON and SOC had a significant linear relationship with the initial concentrations of SON and SOC added into soils. The partition coefficient, m of the initial mass isotherm model, indicated that Argosols had a higher adsorbility than Anthrosols. The average adsorption rates of SON and SOC in Anthrosols were 24.3% and 18.8%, and those in Argosols were 38.3% and 18.6%, respectively. The low adsorption rates of SON and SOC indicated their high mobility in the two soils, and more SON was adsorbed than SOC suggested the higher potential of SOC leaching from soil. PMID- 18419076 TI - [Effects of increasing field temperature on photosynthetic characteristics of summer maize]. AB - A field experiment was conducted to study the effects of increasing field temperature on the grain yield and photosynthetic characteristics of summer maize cultivars ND108 and YD13. The results showed that under increasing field temperature in the period from male tetrad to maturing stage, the grain yield of ND108 and YD13 decreased by 46.6% and 45.1%, leaf area index (LAI) decreased by 15.4% and 11.5%, and net photosynthetic rate of ear leaf decreased by 22.85% and 18.14%, respectively, compared with the control. The contents of leaf chlorophyll a and b, especially that of chlorophyll a, decreased obviously, and the activities of ribulose-1,5-bisphosphate carboxylase (PEPCase) and phosphoenolpyrovate carboxylase (RuBPCase) in the leaves of the two cultivars decreased by 51.1% and 32.4%, and 29.5% and 7.7%, respectively. PMID- 18419077 TI - [Temperature differences of air-rice plant under different irrigated water depths at spiking stage]. AB - With rice cultivars Yangdao 6, Yangjing 9538 and Wuxiangjing 14 as test materials, field experiment was conducted to study the effects of 3 irrigated water depths (0 cm, 2-4 cm, and > 10 cm) on the temperature of different parts of rice plant at spiking stage. The results showed that from 10:30 to 15:00 on sunny days, irrigated water depth on paddy field had significant effects on the temperature of field surface, middle part of rice plant, and rice spike. The higher the water depth on field surface, the lower the temperature of rice plant and rice spike. At the water level > 10 cm, the average temperature differences between air and the rice spike, middle part of rice plant and field surface of these three cultivars were 1.37, 2.98 and 4.12 degrees C higher than those at the water depth of 0 cm, and 0.67, 1.59 and 2.17 degrees C higher than those at the water depth of 2-4 cm, respectively. In addition, the temperature differences were 0.71, 1.39 and 1.95 degrees C higher at the water depth of 2-4 cm than those at the water depth of 0 cm, respectively. Obvious temperature differences of air rice plant were also observed among the three rice varieties under different irrigated water depths. The analysis of the characteristics of temperature transfer among field surface, middle part of plant and rice spike indicated that the temperature transfer patterns under all test water management regimes accorded with the principles of energy transfer, suggesting that keeping proper water depth on the field surface at rice spiking stage contributed great to the decrease of rice spike temperature and the alleviation of rice heat injury. PMID- 18419078 TI - [Effects of mono- and mixed culture on the grain yield and water use efficiency of two winter wheat cultivars]. AB - Taking two winter wheat (Triticum aestivum L.) cultivars Changwu 135 and Pingliang 40 commonly cultivated in the semi-arid area on Loess Plateau as test materials, and by the method of ecological replacement, a 2-year field experiment was conducted to study the effects of mono- and mixed culture on the grain yield and water use efficiency of the cultivars. The results showed that under mono culture, Pingliang 40 had a much higher unit area root biomass (367.60 g x m(-2)) than Changwu 135 (297.31 g x m(-2)), and a more uniform root distribution (i.e., a better root type for water absorption), but its grain yield and water use efficiency were lower than Changwu 135. Under mixed culture, the population root biomass of Pingliang 40 and Changwu 135 was 13.36 g * m(-2) and 8.50 g x m(-2) higher than that under mono-culture, respectively, suggesting that mixed population could absorb the water in deeper soil layers, and had higher total unit area biomass, which in turn increased the water use efficiency. Comparing with Pingliang 40, Changwu 135 allocated more dry matter to its productive organ, leading to its higher grain yield, harvest index, and water use efficiency. PMID- 18419079 TI - [Effects of different fertilization modes on soil ammonia volatilization and nitrous oxide emission]. AB - With enclosed and static chambers, this paper studied the effects of different fertilization modes, i.e., top-dressing nitrogen (N) fertilizer followed by tillage (SF), drilling N fertilizer followed by covering soil (TF), and top dressing N fertilizer followed by irrigation (SS), on soil ammonia (NH3) volatilization and nitrous oxide (N2O) emission. The results showed that fertilization mode had significant effects on the NH3 volatilization and N2O emission. SS promoted NH3 volatilization obviously, with the maximum NH3 volatilization rate higher than other treatments and the total amount of cumulative NH3 volatilization up to 2.465 kg N x hm(-2). There were significant differences (P < 0.05) in the N2O flux among different treatments, and the peak appeared at different time. The N2O flux in SS got to its peak (193.66 miccrog x m(-2) x h(-1)) after two days of fertilization, while that in TF got to the peak (51.13 microg x m(-2) x h(-1), the lowest among different treatments) after five days of fertilization. The net cumulative N2O emission in SS was 121.55 g N x hm( 2), much higher than that of SF and TF. SF and TF reduced NH3 volatilization and N2O emission markedly, suggesting that both of them could be the rational and practicable N fertilization modes. PMID- 18419080 TI - [Estimation of dry matter accumulation in above-ground part of cotton by means of canopy reflectance spectra]. AB - Through analyzing the relationships of the dry matter accumulation in above ground part of cotton with the canopy reflectance of single waveband and all two band combinations in ratio vegetation index (RVI, R(lamda1)/R(lamda2)), normalized difference vegetation index (NDVI, (R(lamda1)-R(lamda2))/(R(lamda1) + R(lamda2 and differential vegetation index (DVI, R(lamda1)-R(lamda2)), the characteristic spectral wavebands for indicating the dry matter accumulation in above-ground part of cotton were determined, and the corresponding prediction model was established. The results showed that the vegetation indices comprised of visible light (560 and 710 nm) and near infrared light (810, 870, 950, 1100 and 1220 nm) were highly related to the dry matter accumulation in the above ground part of cotton, and the RVI (1100, 560) was the best spectral index for the estimation. The corresponding prediction model established by stepwise regression method was Y (g x m(-2)) = 66.274 x RVI (1100, 560)-148.84. It could be feasible to estimate the dry matter accumulation in above-ground part of cotton with remote sensing. PMID- 18419081 TI - [Effects of nitrogen application rate on soil enzyme activities in wheat rhizosphere]. AB - This paper studied the effects of nitrogen application rate on the soil enzyme activities in the rhizosphere of wheat cultivars Lankaoaizao 8, a large spike genotype, and Yumai 49-198, a small spike genotype, under high yield condition. The results showed that the enzyme activities in rhizosphere soil had similar changing trends with wheat growth. The protease, urease and dehydrogenase activities in rhizosphere soil increased with wheat growth, maximized at heading stage, jointing stage, and heading stage, respectively, and decreased thereafter. Catalase activity increased with wheat growth, and peaked at maturing stage. At the same growth stage, the protease, catalase and dehydrogenase activities in rhizosphere soil of the two cultivars increased with increasing nitrogen application rate and peaked at 180 kg N x hm(-2). Urease activity also increased with increasing nitrogen application rate, and the maximum activity was observed at 360 kg N x hm(-2). PMID- 18419082 TI - [Effects of straw mulching on CO2 flux in wintry fallow paddy field]. AB - This paper studied the effects of straw mulching on the CO2 flux in a wintry fallow paddy field at Taoyuan Agro-ecological Station, Chinese Academy of Sciences. The results showed that the effects of straw mulching mainly exerted in two ways. First, it positively affected soil temperature, making the CO2 flux increased obviously. Straw mulching gave a net emission of 2.68 g CO2 x m(-2) x d(-1), while no mulching gave a net fixation of 1.99 g CO2 x m(-2) x d(-1), the difference between them being very significant (P < 0.01). Second, straw mulching decreased the biomass of weeds and the photosynthetically active radiation they absorbed, which in turn resulted in an increase of CO2 flux. Under straw mulching, the water content in surface soil layer (0-15 cm) increased by 9% or more, but no significant change was observed in CO2 flux. PMID- 18419083 TI - [Effects of urbanization on supply and demand of regional ecological footprint]. AB - Based on the time series of ecological footprint (EF) in Jilin Province from 1994 to 2003, the relationship models of EF, ecological budget, and EF intensity with urbanization level were established. The results showed that in Jilin Province, there existed significant correlations of EF, ecological budget, and EF intensity with urbanization level. Along with the development of urbanization, the EF in the Province increased from 1.59 hm2 x cap(-1) in 1994 to 2.23 hm2 x cap(-1) in 2003, which was mainly affected by the process of urbanization and the proportion of tertiary industry. The EFs of built-up land, pasture and fossil fuel land changed more markedly, among which, the EFs of pasture and fossil fuel land were mainly affected by domestic consumption, while that of built-up land was mainly affected by the GDP per capita and the proportion of tertiary industry. Owing to the increase of domestic consumption, the ecological deficit increased from 0.319 hm2 cap in 1994 to 0.923 hm2 cap(-1) in 2003. The changes in ecological budget of pasture and fossil fuel land were more remarkable. Under the effects of the optimization of economic structure and consumption structure, the EF intensity in the Province decreased from 4.14 hm2 x (10(4) Yuan)(-1) in 1994 to 2.35 hm2 (10(4) Yuan)(-1) in 2003, and there still had enough potential for the decrease. Through the optimization of economic structure and consumption structure, an ecological surplus and the balance between natural resources supply and demand in the Province could be achieved. PMID- 18419084 TI - [Quantitative monitoring of gully erosion in hilly-gully area of Loess Plateau based on aerial images]. AB - Based on the aerial photographs in 1979 and 1993, and by using geographical information system (GIS) techniques, the digital elevation models (DEM) and digital orthophoto map (DOM) were constructed from the geomorphic changes in a chosen gully in the Loess hilly-gully area, and the spatial difference of the changes and their key affecting factors were analyzed. The results indicated that from 1979 to 1993, the gully head advanced actively (with a length of 8.5 meters and an erosion rate of 481.28 t x hm(-2) x a(-1)), and the slope tended to become steeper, exhibiting a high erosion risk. The middle part of the gully had the largest newly eroded area and the highest erosion rate, and the erosion process was quite active, with the highest erosion risk. The water outlet was primarily endowed with deposition formation, whose eroded area was smaller than its deposition area, and the gully depth and slope were becoming less, with a lower erosion risk. PMID- 18419085 TI - [Early warning and risk assessment of snow disaster in pastoral area of northern Xinjiang]. AB - Based on the analysis of animal husbandry production and of distribution characteristics of snow disaster in northern Xinjiang, and by using RS and GIS techniques and field survey data, 9 early warning factors were selected from the three subsystems of grassland' s disaster-resistant capability, livestock's disaster-bearing capacity, and disaster-causing potential, and the death rate of livestock caused by snow disaster was used as a factor of risk assessment. An index system of snow disaster' s early warning and risk assessment for completely grazing grassland was established, and the early warning model of snow disaster, its distinguishing model, and risk assessment model were built by using multi hierarchical synthetic and multi-objective linear weight function methods to predict the resistant capability of grassland and livestock against snow disaster, and to assess the potential risk loss from snow disaster in northern Xinjiang. The accuracy of the early warning model and risk assessment model was 85% and 72% , respectively. PMID- 18419086 TI - [Limitations of landscape pattern analysis based on landscape indices: a case study of Lize-jian wetland in Yilan of Taiwan Province, China]. AB - In this paper, the Lizejian wetland landscape patterns in northeastern Taiwan of China were established by landscape indices and aerial photo interpretation, and a parallel analysis was made on them. The results showed that landscape indices could only indicate the landscape geometric characteristics of the wetland at patch and landscape levels, but could not present its spatial and functional characteristics observed from aerial photos. Combining aerial photo interpretation with landscape indices could be helpful to the holistic understanding of Lizejian wetland' s landscape structure and function, and improve the landscape pattern analysis. The new method for assessing landscape structure from a holistic point of view would play an important role in future landscape ecology research. PMID- 18419087 TI - [Diet composition and feeding habits of juvenile Acipenser sinensis in Yangtze River estuary]. AB - This paper dealt with the diet composition and monthly variation in feeding habits of juvenile Acipenser sinensis accidentally caught along with other fish in Yangtze River estuary from May to September, 2004. The results showed that the juvenile A. sinensis mainly fed on demersal fish, polychaeta and amphipoda, as well as some small-sized benthic invertebrates including shrimp, crab and bivalve. The feeding rate and intensity were higher, and the feeding intensity varied monthly. The main food items also varied monthly, and changed from small sized to relative large-sized species. The main diet composition included polychaeta and crab in May, fish and amphipoda in June, fish and polychaeta in July, empty stomach in August, and fish and shrimp in September. In the diet, the proportion of Platycephalidae, an important prey of juvenile A. sinensis in the past, was very low, only accounting for 1.81% of the total food by mass, while that of Gobiidae and Isopoda, which had not been recorded in the diet, was higher, accounting for 50.54% and 10:07% of the total, respectively. PMID- 18419088 TI - [Influences of Solenopsis invicta buren invasion on the native ant communities in different habitats in Guangdong]. AB - By using pitfall and bait traps, an investigation was made on the diversity and similarity of ant communities in the areas infested and un-infested with Solenopsis invicta Buren in Shenzhen of Guangdong. The results showed that under the invasion of S. invicta, the ant species number in lawn and wasteland reduced obviously, with a decrease of 6 in lawn and 3 in wasteland, and the native dominant ant species in lichee orchard, especially in wasteland and lawn, were replaced by S. invicta. With the infestation of S. invicta, the diversity and evenness of ant communities in wasteland and lawn decreased but the predominance increased obviously, while it was in adverse in lichee orchard. The similarity coefficients of the ant communities between S. invicta infested and un-infested lichee orchard, wasteland and lawn were 0.6316, 0.5882 and 0.2941, respectively. PMID- 18419089 TI - [Effects of methamidophos and 17beta-estradiol on the dynamics of experimental population of freshwater rotifer Brachionus calyciflorus]. AB - The study on the dynamics of experimental population of freshwater rotifer Brachionus calyciflorus under exposure of different concentrations of methamidophos (0.01, 0.1, 1.0, 10.0, 100.0, 1000.0 and 10000.0 microg x L(-1)) and 17beta-estradiol (0.001, 0.01, 0.1, 1.0, 10.0, 100.0 and 1000.0 microg x L( 1)) showed that both methamidophos and 17beta-estradiol had significant effects on the mean growth rate of the population, but less affected the mean fertilization rate of mictic females. Methamidophos affected the mean ratio of ovigerous to non-ovigerous females and the mean mictic rate significantly but had less effect on the total resting egg production, while 17beta-estradiol was in adverse. Compared with the controls, both 10.0-10000.0 microg x L(-1) of methamidophos and 100.0 microg x L(-1) of 17beta-estradiol increased the mean population growth rate significantly. 0.1-10000.0 and 10.0 microg x L(-1) of methamidophos decreased significantly the average ratio of ovigerous to non ovigerous females and the average mictic rate, respectively, and 1000.0 microg x L(-1) of 17beta-estradiol decreased the total resting egg production significantly. Within the range of test concentrations of methamidophos, the relationship between the mean population growth rate (Y, d(-1) and the concentration (X, microg x L(-1) of methamidophos could be described as Y = -2 x 10(-8) X2 + 0.002X + 0.374. The population growth rate could be used to assess the effects of methamidophos on the population dynamics of the rotifer. PMID- 18419090 TI - [Community structure characteristics of zooplankton in Yili region of Xinjiang in summer]. AB - In June-July 2006, investigations were made on the zooplankton in the mainstream of Yili River and its three main tributaries (Tekes River, Kunes River and Kax River), two reservoirs (Qiapuqihai Reservoir and Jilintai Reservoir), intermittent waters, and ponds in Yili region of Xin-jiang. A total of 217 species belonging to 134 genera were identified, among which, Protozoa and Rotifer were the main groups, which contained 153 species (71% of the total species number) of 95 genera and 51 species (23% of the total) of 27 genera, respectively. Only 8 species of Cladocera and 5 species of Copepoda were observed. The zooplankton richness ranged from 7 to 132, with the lowest (7) in Jilintai Reservoir, and the highest (132) in Yili River. Qiapuqihai Reservoir had the highest density and biomass of zooplankton (11391 ind x L(-1) and 2.79 mg x L(-1), respectively), but Tekes River had the lowest density (578 ind x L(-1)) and Kax River had the lowest biomass of zooplankton (0.03 mg x L(-1)). Pearson correlation analysis and linear regressions showed a significantly negative relationship between zooplankton species number and water flow velocity. Based on the species number, density, and biomass of zooplankton, it was found that Yili River and its three main tributaries were in oligotrophic status, while Qiapuqihai Reservoir and Jilintai Reservoir were in mesotrophic status. PMID- 18419091 TI - [Effects of wheat synomones on Macrosiphum avenae and its natural enemies]. AB - An investigation was made in winter wheat fields to study the effects of applying wheat synomones methyl salicylate and sulcatone (6-methy-5-hepten-2-one) on the dynamic changes of Macrosiphum avenae and its natural enemies Harmonia axyridis and Aphidius avenae. The results showed that wheat synomones didn' t change the population dynamics of M. avenae significantly, but had a significant effect on the reduction of M. avenae population number. The synomones could deteriorate the existent environment of M. avenae, and make the number of alatae increased obviously. Although synomones decreased the species richness, diversity index, and evenness of natural enemy communities, the numbers of dominant natural enemy species such as A. avenae and H axyridis were increased. Applying wheat synomones was helpful to the control of M. avenae in winter wheat fields. PMID- 18419092 TI - [Utilization and management of Trichiurus japonicus resources in East China Sea based on Beverton-Holt model]. AB - In this paper, the population structure of Trichiurus japonicus in East China Sea was analyzed based on the data of its age composition and anus length in 2002 2004, and the rational utilization of T. japonicus resources in the Sea was approached by using the catch (in mass) per recruit theory of Beverton-Holt model. The results revealed that: (1) there was a miniaturization trend of the T. japonicus population in East China Sea. The range of age composition changed from 0-6 year in the late 1950's to 0-4 year in the early 21st century, and the population was dominated by the group of 0-1 year now. The percentage of 2-year old T. japonicus decreased from 12.84% in the late 1950s' to 6.91% in the early 21st century, and that of 3-year old T. japonicus decreased from 4.92% in the late 1950s' to 0.57% in the early 21st century; (2) the exploitation rate of T. japonicus in the period of 2000-2003 was 0.864, which was beyond of the optimum exploitation rate of 0.51, suggesting that the T. japonicus in East China Sea was under over-fishing; and (3) to enhance the age value at first capture (t(c)) was the best measure of increasing the catch (in mass) per recruit of T. japonicus. The age at recruitment (t(r)) and the t(c) of T. japonicus in East China Sea was 0. 25 and 0.5 at present, respectively. If the t(c) changed from 0.5 to 1, the unit catch would be increased by 55.38%, and if the t(c) changed to 1.5, 2 or 2.75, it would be increased by 100.81%, 130.52% or 145.23%, respectively. It was suggested that due to the difficulties in greatly reducing catching intensity, it could be available to properly increase the to value while decrease the catching intensity to protect the T. japonicus resources in East China Sea and realize their sustainable use. PMID- 18419093 TI - [Research progress on plant diversity conservation in sand dune areas]. AB - The landscape in sand dune areas is characterized by the alternate distribution of sand dune and interdune lowland, and the unique floras in these areas are some endemic or rare plant species. In recent years, the decrease in plant species richness and the disappearance of some endemic or rare plant species in these areas have been received special attention, which were listed in the Program of International Biodiversity Conservation, and studied in many countries and districts. In this paper, the research progress in these fields was summarized from the aspects of significance of plant diversity conservation, formation mechanisms of plant diversity, ways of plant diversity conservation, roles of plant diversity research in the development of ecological theories, and important issues in operating plant diversity conservation project. To conserve plant diversity in sand dune areas, attentions should be paid to the differences in conservation goals (to maintain high species richness or to conserve endemic or rare species) among different regions, and the balances between the stabilization of active sand and the conservation of endemic or rare species, and the maintenance of high species richness and the conservation of endemic or rare species. It needed also to consider the sand dune and the interdune lowland as a unified landscape unit to explore the impacts of disturbances and habitat fragment on plant diversity. PMID- 18419094 TI - [Relationships between seed size and seedling growth strategy of herbaceous plant: a review]. AB - Seed size and seedling recruitment strategy are of importance in the life-history strategy of plant. In this paper, the current ecological researches at home and aboard on the relationships between seed size and seedling growth were reviewed from the aspects of the effects of seed size on seed germination and seedling emergence, the relationships between seed size and seedling growth traits, and the relationships between seed size and seedling survival and competition ability. Some suggestions on future researches in this field were put forward. There were likely different relationships between seed size and seedling growth in different microenvironments and vegetation types, and the effects of seed size on seedling growth could result in different contributions of different seed-size species to the seedling recruitment of vegetation. The large-scale community level and the small-scale intra- and inter-species level researches on this issue should be strengthened, which would have significance for the recruitment and renewing of natural vegetation. PMID- 18419095 TI - [Effects of abiotic factors on algal extracellular polysaccharides content]. AB - To get insight of the phenotypic variation mechanisms of Microcystis aeruginosa, this paper reviewed the research results on the effects of various abiotic factors on the content of algal extracellular polysaccharides. Many published papers demonstrated that there exists a definite responsive relationship between the content of algal extracellular polysaccharides and the ratios of main nutritive elements in the environment. Under the condition of high C:N or C:P ratio, i.e., under nitrogen or phosphorus deficiency, the organic substances fixed by the photosynthesis of algae are mainly existed in the forms of carbohydrates that do not contain nitrogen and phosphorus. The excessive accumulation of polysaccharides in algal cell induces their gradual release to the outside, resulting in a significant increase in the content of extracellular polysaccharides. Under the unbalanced condition of C or N metabolism, the extracellular polysaccharides can serve as a sink of excessively fixed carbon. For several kinds of algae, different light quality, light intensity and photoperiod can affect their synthesis and secretion of extracellular polysaccharides, and temperature can also affect the production of their extracellular polysaccharides to a certain extent. As the extracellular polysaccharides play a very important role in cementing cells together to form colonies, it's speculated that the increase of extracellular polysaccharides production by M. aeruginosa via the regulation of relevant abotic factors could be helpful to simultate the colony formation of M. aeruginosa in laboratory. PMID- 18419096 TI - [Relationships between landscape pattern and water quality at western reservoir area in Shenzhen City]. AB - In this paper, the relationships between landscape pattern and water quality at western reservoir area in Shenzhen City were studied with grey connection method, and the influences of source' and 'sink' landscape patterns on non-point pollution were probed. The results showed that the dominance, adjacency, and fragmentation of 'source' and 'sink' landscapes could markedly influence the water quality. From 2000 to 2001, due to the changes of the 'source' and 'sink' landscape patterns in research areas, the output of pollutants increased and the reduction of pollution decreased, resulting in the deterioration of water quality of three reservoirs. According to the spatial distribution of 'source' and 'sink' landscapes, it was found that the distribution of 'sink' landscapes at the middle lower reaches of the watersheds had close relationships with the changes of reservoir water quality, suggesting that 'sink' landscape pattern was of significance in the management of non-point pollution. PMID- 18419097 TI - [Changing characteristics of temperature and humidity in different type bags for bagging apple and their effects on fruit appearance quality]. AB - This paper studied the changing characteristics of temperature and humidity in different type bags for bagging Fuji apple during its development and their effects on the appearance quality of the fruit. The results showed that there was a negative correlation between the changes of temperature and humidity in the bags. Temperature started to rise rapidly from 9:00, with the highest appeared generally from 13:00 to 14:00 in spring and autumn or from 13:00 to 17:00 in mid summer, while the lowest at about 5:00. In parallel, the highest humidity usually appeared at about 6:00. There were different microenvironments in different bags, especially in hot and rainy days. The highest temperature appeared in double layer bags, while the highest humidity appeared in single-layer ones. Usually, high temperature led to extremely dry, e.g. the humidity in bags was below 40% at 35 degrees C. Bagging had obvious effects on fruit quality. Comparing with single layer bags, double-layer bags had better effects in improving fruit appearance quality, and the best was that of type XL. PMID- 18419098 TI - [Mineral nutrition niche of rice sheath blight fungus]. AB - By using the principles and methods of niche, this paper studied the mineral nutrition niche of rice sheath blight fungus. The results showed that the mineral nutrition niche breadth of the fungus at rice tillering, booting, heading, and waxing stages was 0.2710, 0.3865, 0.4252 and 0.4817, respectively, i.e., increased with rice growth, but still comparatively narrower, indicating that rice sheath blight fungus only utilized smaller parts of mineral nutrients at various growth stages of rice. The fungus always gave priority to occupy the nutrition niches where Mg, Zn and Si contents were lower, suggesting that Mg, Zn and Si contents had close relations to the resistance of rice against rice sheath blight. PMID- 18419099 TI - [Role of earthworm in degradation of soil phenanthrene by Pseudomonas putida]. AB - A 30-day incubation test was conducted to investigate the effects of treatments earthworm (E), bacteria (Pseudomonas putida) (B) and earthworm-bacteria (BE) on the degradation of soil phenanthrene. The degradation rate of soil phenanthrene at its initial concentration of 50 mg x kg(-1) was in the sequence of BE > B > E > CK, and that at the concentration of 150 mg x kg(-1) was 98.86% in BE, being significantly higher than that in CK and E. With the increase of the initial concentration of soil phenanthrene, the bacterial dioxygenase activity almost did not change in B, but increased significantly in BE. Under the same concentration of soil phenanthrene, the phenanthrene content in earthworm was significant higher in BE than in E, suggesting that earthworm could decrease the concentration of soil phenanthrene via its bioaccumulation, and the interaction between earthworm and P. putida could further promote the biodegradation of soil phenanthrene. PMID- 18419100 TI - [Guidelines of reference recording in scientific papers of Chinese Journal of Applied Ecology]. AB - To improve the compilation quality of references, work in well with articles search and periodicals evaluation, and promote international academic exchange, the Chinese Journal of Applied Ecology shall adjust its principles of reference recording in scientific papers based on the GB/T7714 -2005. From 2008, the references in scientific papers to be submitted are requested to record by the Citation-Sequence. In this paper, some examples were presented, and the issues needed to be paid more attention to by the authors were put forward. PMID- 18419101 TI - [Applying the DNA diagnostics in patients with superficial keratitis of viral origin]. AB - AIM: The authors evaluate the significance of the DNA diagnostics in patients with superficial keratitis of viral origin and their capability to be used for monitoring of the treatment effectiveness in the follow-up. The presence of herpes simplex virus 1 and 2, varicella zoster virus (HSV 1, HSV 2, VZV), and adenoviruses was assessed by means of the DNA analysis. MATERIAL AND METHODS: The group consisted of 54 patients (33 men and 21 women), mean age 45.6 +/- 9.5 years, who were treated at the Eye Department for superficial keratitis or keratouveitis of viral origin. A sample from the involved place was taken with a cotton swab and a sample of approx. 50 microL of tears was taken from the conjunctival sac with a single-use micropipette. The cotton swab and the tears were shaken with the EliDNA Store Kit (ELISABETH PHARMACON, Czech Republic) buffer, which prevents the DNA degradation and allows the storage and transport of samples at the room temperature. After the transportation in to the laboratory, the DNA was isolated by means of the UltraClean DNA Tissue Kit (MoBio, U.S.A.). The isolated DNA was used for HSV 1, HSV 2, VZV, and adenoviruses detection by means of PCR (polymerase chain reaction). All samples were screened for the HSV1 presence using the in-house RealTime method with TaqMan probe and the Applied Biosystems RealTime System 7300 device. In case of positive result of the DNA analysis, control samples were taken in 7 - 10 days periods until negative result was obtained; another sample was taken in case of suspected relapse. The control examination was also performed by means of cultivation from the same sample by another laboratory. RESULTS: Altogether 82 samples were taken and 230 DNA analyses were performed. The DNA diagnostics proved the presence of HSV 1 DNA in 28 patients, in one case, VZV DNA was detected, and 16 patients were positive on adenoviruses. The HSV 1 positive samples were confirmed by means of in-house RealTime PCR method as well as commercially available in vitro diagnostic healthcare device End-Point PCR HSV1/2 (Nanogen Advanced Diagnostics, Italy). All cultivation control examinations performed in another laboratory were negative. The samples were taken repeatedly in 9 patients to monitor the efficacy of the treatment. SUMMARY: The DNA diagnostics seem to be a fast and reliable method to determine the etiological agent in patients with superficial keratitis and allow very accurate monitoring of the treatment efficacy. PMID- 18419102 TI - [The application of the autologous serum eye drops results in significant improvement of the conjunctival status in patients with the dry eye syndrome]. AB - AIM: To detect the changes on the conjunctiva surface before and after the application of the autologous serum (AS) eye drops in patients with dry eye syndrome, using both clinical and laboratory approaches, supplemented with subjective assessing the discomfort status. MATERIALS AND METHODS: The AS eye drops were applied during the period of 3 months in 8 patients with dry eye syndrome (Schirmer test < 5 mm and break-up time < 5 seconds), with the highest (maximum) frequency 8 times a day. The clinical (Schirmer test, break-up time, rose Bengal staining, examination of the tear meniscus, detritus and superficial punctate keratitis) and laboratory examinations (morphological assessment of the conjunctiva, detection of apoptotic cells) were performed at the start and at the end of the 3 months treatment period. Each day, patients reported their ocular status (dryness, discomfort, foreign body sensation, light sensitivity). RESULTS: The AS eye drops application improved significantly the values of the Schirmer test, detritus and superficial punctate keratitis as well. The goblet cells density on the conjunctival surface increased and the number of apoptotic cells decreased. The intensity of unpleasant feelings reported by the patients decreased significantly in all of the assessed categories. CONCLUSION: Because the application of AS eye drops caused the improvement of conjunctival status as well as the decrease of the severity of difficulties reported by the patients, the AS eye drops application should become common therapeutic practice in patients with dry eye syndrome. PMID- 18419103 TI - [Arteriovenous decompression for branch retinal vein occlusion with internal membrane peeling for macular edema]. AB - PURPOSE: A retrospective study of anatomical and functional results of microsurgical therapy of branch retinal vein occlusion with internal limiting membrane peeling due to macular edema. MATERIALS AND METHODS: Eleven patients (5 men and 6 women), mean age 59.18 years (34-74 years) who underwent the surgery at the Department of Ophthalmology, Comenius University in Bratislava, Slovak Republic, from June 1st, 2000 to May 31st, 2006, were enrolled in the study. The follow-up period ranged from 14 to 72 months (average 28.5 months). The patients were indicated to the arterio-venous decompression with internal retinal membrane peeling due to the macular edema after the fluorescein angiography. Patients with rubeosis of the iris were excluded from further evaluation and their initial best corrected visual acuity was 0.3 and less. A complete eye examination (best corrected visual acuity, intraocular pressure, fluorescein angiography, slit lamp examination of both the anterior and posterior eye segments) was performed in each patient before the surgery, and every 3-4 months during the first year and every 6-9 months during following years. Fluorescein angiography was used to evaluate anatomic results 3 months after the surgery. As positive signs were considered: the blood flow improvement peripherally to the arteriovenous decompression site, the vessel dilatation, and the reduction of the hyperfluorescence and the macular edema reduction. To analyze the functional results, the best corrected visual acuity changes before and after surgery were used. RESULTS: Positive anatomic changes were in 8 (72,73%) patients; in 1 patient the area of non-perfusion expanded, and 2 patients were presented with stable anatomic findings. The best corrected visual acuity averaged 0.16 (+/- 0.1070) before the surgery, 0.2909 (+/- 0.2264) 3 months after the surgery, and 0.3818 (+/- 0.3178) at the time of the last examination. Significant differences between the visual acuity before and 3 months after surgery were confirmed by Student's T-test. The visual acuity measured at the time of the last measurement improved by +2 and more lines in 6 patients, reminded unchanged in 4 patients, and worsened by -2 or more lines in 1 patient. CONCLUSION: Microsurgical treatment of branch retinal vein occlusion expands the therapeutic armamentarium used to manage this severe disease. In case the partial occlusion is present, arteriovenous decompression has a positive effect on the final anatomic and functional results. PMID- 18419104 TI - [Spontaneous premacular hemorrhage]. AB - PURPOSE: Among the great amount of etiologically heterogeneous group of general and specific eye diseases, a solitary, spontaneous, unilateral retinal hemorrhage restricted to the macula in young adults can be regarded as an entity. METHODS: Twelve eyes were phakic, emmetropic (including minor non corrected astigmatic errors) and were seen over a period of 5 years. All patients had a complete ophthalmologic examination with special analysis of history of direct or indirect trauma, additional medical work-up to exclude hematological disorders, and fluorescein angiography examinations up to the complete blood resolution. RESULTS: Hemorrhage is presented in 3 pictures: 1) Foveal microhemorrhage, 2) Drop-like hemorrhage limited to the avascular zone of the fovea, 3) Bag-like hemorrhage over the posterior pole. Initially, all patients had complains suggestive of positive central or paracentral scotomas with impaired visual acuity. The resolution of blood was spontaneous in 10 patients, after argon laser treatment in 2 patients. In all of them, the recovery of the visual acuity was complete. Possible pathogenetic mechanism such as Valsalva manoeuvre could be found in 3 cases only. CONCLUSIONS: Spontaneous macular hemorrhage is mostly idiopathic clinical entity in young adults with complete recovery of visual acuity. PMID- 18419105 TI - [Treatment for recurrent pterygium]. AB - Our objective was to evaluate the postoperative outcomes of the surgical treatment for recurrent pterygia. Twelve eyes of 10 patients operated on for recurrent pterygia at the Department of Ophthalmology, University Hospital Hradec Kralove, Czech Republic, were involved in this study. All surgeries were done under the local anesthesia. The pterygium head was undermined and removed by dissection to reach the clear corneal lamelae. The pterygium body was thoroughly removed with extensive dissection of scared pathological tissue. In 3 eyes, the Mitomycin C 0.004% solution was applied. Preserved human amniotic membrane was placed over the corneal and conjunctival defects and sutured with 8/0 Vicryl interrupted sutures. Postoperative results were good; in 11 eyes we did not observe the true corneal recurrence of fibrovascular tissue. In one eye only, the lamellar sclerokeratoplasty was necessary to perform because of the recurrence of pterygium. Recurrent pterygium exhibits a more aggressive, fibrovascular growth pattern, leading to the corneal and conjunctival scarring and limbal stem cells deficiency. Proper excision of pathological tissue with amniotic membrane transplantation and/or Mitomycin C represents an alternative surgical method with good final outcomes. PMID- 18419106 TI - [GDx before and after LASIK in middle and high myopia]. AB - AIM: To evaluate, if there are statistically significant changes in the RNFL (retinal nerve fibre layer) after LASIK. To evaluate, if the changes in the corneal structure caused by LASIK involve the results of the RNFL by means of GDx VCC. MATERIAL AND METHODS: The group consisted of 100 eyes of 51 patients (32 women, 19 men); the mean age was 28.55 +/- 5.1 years (18-50 years). The average refractive error in spherical equivalent (SE) was -5.46 +/- 1.40 D (dioptres). The group was divided into two subgroups: subgroup A (69 eyes with SE from -3.25 D to -6.0 D), and subgroup B (31 eyes with SE from -6.25 D to -12 D). The patients underwent the LASIK procedure to correct the myopia. The thickness of the RFNL was measured by means of GDx analyzer with variable corneal compensator. The measurements were performed before and 1, 3, 6, and 12 months after the LASIK procedure. The results of the measurements were statistically evaluated by means of the Mann-Whitney U test. RESULTS: The statistically significant difference in the RNFL thickness (p < 0.05) was found in "Superior Average" 3 and 12 months after LASIK (p = 0.016, p = 0.018), in "Inferior Average" in all controls (p = 0.047, p = 0.0001, p = 0.0003, p = 0.001) and in "NFI" after 12 months (p = 0.039). The values of difference in RNFL thickness in separate measurements after LASIK between both subgroups A and B were evaluated by means of Mann-Whitney U nonparametric test. Statistically significant difference in "Inferior Average" was found 1,6, and 12 months after LASIK (p = 0.01, p = 0.01, p = 0.04); in "TSNIT Average" after 6 months (p = 0.01); in "NFI" values after 1 month (p = 0.03). In "Superior Average, no statistically significant difference was found. SUMMARY: In our group we have found statistically significant decrease of RNFL thickness after LASIK in every single quadrant. Clinically, the differences in RNFL thickness before and after LASIK were minimal. We suppose the measurements by means of GDX are influenced by changes in the polarization features of the cornea caused by LASIK procedure. PMID- 18419107 TI - [The presence of dry eye syndrome and corneal complications in patients with rheumatoid arthritis and its association with -174 gene polymorphism for interleukin 6]. AB - The aim of this study was to evaluate the presence of the dry eye syndrome and corneal complications in patients with rheumatoid arthritis and to assess its association with the -174 gene polymorphism for interleukin 6. The group consisted of 123 patients treated for rheumatoid arthritis (20 men, 103 women); the mean age was 53 years (+/- 13.6). Every patient had completely ophthalmologic examination and special attention was paid to the amount of tears. The presence of corneal complications was detected in the medical history and evaluated during the slit lamp examination. In all patients the polymorphism-174 IL-6 examinations were performed. For the statistical data processing, the chi square (chi2) test for nominal variable was used. The dry eye syndrome (DES) was found in 98 eyes (79.7%), severe dry eye syndrome was detected in 53 patients (43.1%). Corneal complications appeared in 9 patients (7.3%). DES was present in 32 patients with the GG genotype (91.4%, n1 = 35), in 49 patients with the CG genotype (71.0%, n2 = 69), and in 8 patients with the CC genotype (42.1%, n3 = 19). After the statistical evaluation we have found the association between the dry eye syndrome and the GG genotype (chi2 = 8.9) and the association between less common dry eye syndrome appearance and the presence of the CC genotype (chi2 = 10.3). Severe dry eye syndrome we proved in 18 patients with GG genotype (51.4%, n1 = 35), in 31 patients with CG genotype (44.9%, n2 = 69), and in 4 patients with CC genotype CC (21.1%, n3 = 19). We proved statistically significant association between CC genotype and less often appearance of the severe dry eye syndrome (chi2 = 4.45). Corneal complications we noticed in one patient with GG genotype (2.8%, n1 = 35), in 5 patients with CG genotype (7.2%, n2 = 69), and in 3 patients with CC genotype (15.8%, n3 = 19). We did not prove statistically significant association between the 174 IL-6 polymorphism and corneal complications appearance. The 174 IL-6 polymorphism influences the appearance of the dry eye syndrome. In patients with GG genotype of the -174 gene polymorphism for IL-6 is its appearance more common. Patients with the rheumatoid arthritis and with CC genotype of the -174 gene IL-6 polymorphism have lower frequency of the dry eye syndrome presence. PMID- 18419109 TI - Benzofuran-based hybrid compounds for the inhibition of cholinesterase activity, beta amyloid aggregation, and abeta neurotoxicity. AB - The complex etiology of Alzheimer's disease (AD) prompts scientists to develop multitarget strategies to combat causes and symptoms. We therefore designed, synthesized, and tested new hybrid molecules linking a benzofuran ring to a N methyl- N-benzylamine through a heptyloxy chain, affording a series of potential multifunctional drugs for AD. The cholinesterase inhibitory activity was extended to the inhibition of Abeta fibril formation for 1, 3, and 5. Compound 3 showed an additional neuroprotective effect. PMID- 18419108 TI - 2-amino-1,3-thiazol-4(5H)-ones as potent and selective 11beta-hydroxysteroid dehydrogenase type 1 inhibitors: enzyme-ligand co-crystal structure and demonstration of pharmacodynamic effects in C57Bl/6 mice. AB - 11beta-hydroxysteroid dehydrogenase type 1 (11beta-HSD1) has attracted considerable attention during the past few years as a potential target for the treatment of diseases associated with metabolic syndrome. In our ongoing work on 11beta-HSD1 inhibitors, a series of new 2-amino-1,3-thiazol-4(5 H)-ones were explored. By inserting various cycloalkylamines at the 2-position and alkyl groups or spirocycloalkyl groups at the 5-position of the thiazolone, several potent 11beta-HSD1 inhibitors were identified. An X-ray cocrystal structure of human 11beta-HSD1 with compound 6d (Ki=28 nM) revealed a large lipophilic pocket accessible by substitution off the 2-position of the thiazolone. To increase potency, analogues were prepared with larger lipophilic groups at this position. One of these compounds, the 3-noradamantyl analogue 8b, was a potent inhibitor of human 11beta-HSD1 (Ki=3 nM) and also inhibited 11beta-HSD1 activity in lean C57Bl/6 mice when evaluated in an ex vivo adipose and liver cortisone to cortisol conversion assay. PMID- 18419110 TI - Thiocamptothecin. AB - The synthesis and the X-ray structure of 16a-thiocamptothecin (TCPT), the thiopyridone analog of camptothecin (CPT), are accomplished. The crystal contains two structurally identical, yet independent molecules. Both of them are connected to other molecules via two intermolecular hydrogen bonds. S-methylation of TCPT leads to the cleavage of the C-ring. The cytotoxic activity of TCPT was evaluated against different human tumor cell lines using CPT as reference compound. PMID- 18419111 TI - Straightforward recursive partitioning model for discarding insoluble compounds in the drug discovery process. AB - Poor aqueous solubility is one of the major issues in drug discovery and development, impacting negatively on all aspects of the research and development process. The pharmaceutical industry has realized that solubility issues need to be resolved at the discovery stage. We here present an innovative way to address this problem via a model designed to address the simple question, "Is the compound likely to be sufficiently soluble to provide interpretable data in biological screening assays?" A recursive partitioning (RP) method was applied to a set of 3563 molecules, with in house determined aqueous solubility values. Five models were generated on the basis of a small number of descriptors affording intuitive information regarding structural features influencing solubility. The final model was based on only two descriptors: the molecular weight (MW) and the aromatic proportion (AP). This model provided satisfactory values of accuracy (81%) and precision (75%) for a test set of 1200 compounds, suggesting that the model may add value in compound selection and library design during early drug discovery. PMID- 18419112 TI - Non-peptide gonadotropin-releasing hormone receptor antagonists. PMID- 18419113 TI - Effects of targeting moiety, linker, bifunctional chelator, and molecular charge on biological properties of 64Cu-labeled triphenylphosphonium cations. AB - In this report, we present the synthesis and evaluation of six new 64Cu-labeled triphenylphosphonium (TPP) cations. Biodistribution studies were performed using the athymic nude mice bearing U87MG human glioma xenografts to explore the impact of TPP moieties, linkers, bifunctional chelators (BFCs), and molecular charge on biological properties of 64Cu radiotracers. On the basis of the results from this study, it is concluded that (1) mTPP (tris(4-methoxyphenyl)phosphonium) is a better mitochondrion-targeting molecule than TPP and 3mTPP (tris(2,4,6 trimethoxyphenyl)phosphonium); (2) DO3A (1,4,7,10-tetraazacyclododecane-4,7,10 triacetic acid) and DO2A (1,4,7,10-tetraazacyclododecane-4,7-diacetic acid) are suitable BFCs for the 64Cu-labeling of TPP cations; (3) NOTA-Bn ( S-2-(4 thioureidobenzyl)-1,4,7-triazacyclononane-1,4,7-triacetic acid) has a significant adverse effect on the radiotracer tumor uptake and tumor-to-background ratios; and (4) monoanionic BFCs should be avoided to ensure that 64Cu chelate has a neutral or negative charge. Considering the tumor uptake and tumor/liver ratios, 64Cu(DO2A-xy-TPP)+ is the best candidate for more extensive evaluations in different tumor-bearing animal models. PMID- 18419114 TI - Probing the structures of leishmanial farnesyl pyrophosphate synthases: homology modeling and docking studies. AB - Leishmania donovani and Leishmania major farnesyl pyrophosphate synthase ( LdFPPS and LmFPPS) are potential targets for the development of antileishmanial therapy. The protein sequence for LdFPPS was recently elucidated in our laboratory. Highly refined homology models were generated using the protein sequences of LdFPPS and the closely related LmFPPS enzyme. A ligand-refined model of LmFPPS with a bound bisphosphonate ligand was generated using restraint-guided molecular mechanics followed by quantum mechanics/molecular mechanics refinement. The ligand-refined model of LmFPPS was further validated through extensive pose validation, enrichment, and other docking studies involving known bisphosphonate inhibitors. The model was able to explain the critical binding site interactions and site directed mutagenesis data obtained from experimental studies on related FPPS enzymes. The ligand-refined model in conjunction with the validated docking protocol could be utilized in the future for structure-based virtual screening and rational drug design studies against these targets. PMID- 18419115 TI - A density functional theory calculation of the electronic properties of several high-spin and low-spin iron(II) pyrazolylborate complexes. AB - Density functional theory has been used to study the electronic spin-state properties of low-spin Fe[HB(pz)3]2, high-spin Fe[HB(3-Mepz)3]2, high-spin Fe[HB(3,5-Me 2pz)3]2, and high-spin Fe[HB(3,4,5-Me 3pz)3]2 complexes that exhibit very different iron(II) electronic spin-sate crossover behaviors with changing temperature and pressure. Excellent agreement is obtained between the experimentally observed Mossbauer-effect quadrupole splittings and isomer shifts of these complexes and those calculated with the B3LYP functional and various different basis sets for both the high-spin and low-spin states of iron(II). The calculations for Fe[HB(pz)3]2 that use the LANL2DZ, 6-31++G(d,p), and 6 311++G(d,p) basis sets for iron all lead to very similar electric field gradients and thus quadrupole splittings. The initial calculations, which were based upon the known X-ray structures, were followed by structural optimization, an optimization that led to small increases in the Fe-N bond distances. Optimization led to at most trivial changes in the intraligand bond distances and angles. The importance of the 3-methyl-H...H-3-methyl nonbonded intramolecular interligand interactions in controlling the minimum Fe-N bond distances and determining the iron(II) spin state both in Fe[HB(3-Mepz)3]2 and in the related methyl substituted complexes has been identified. PMID- 18419118 TI - Reactivity of the triple ion and separated ion pair of tris(trimethylsilyl)methyllithium with aldehydes: a RINMR study. AB - Low-temperature rapid-injection NMR (RINMR) experiments were performed on tris(trimethylsilyl)methyllithium. In THF/Me2O solutions, the separated ion (1S) reacted faster than can be measured at -130 degrees C with MeI and substituted benzaldehydes (k >/= 2 s -1), whereas the contact ion (1C) dissociated to 1S before reacting. Unexpectedly, the triple ion reacted faster with electron-rich benzaldehydes relative to electron-deficient ones. The addition of HMPA had no effect on the rate of reaction of the triple ion with p-diethylaminobenzaldehyde, and the immediate product of the reaction was the HMPA-solvated separated ion 1S, with the Peterson product forming only slowly. Thus, the aldehyde is catalyzing the dissociation of the triple ion. HMPA greatly decelerated the reaction of 1S (<10 -10), providing an estimate of the Lewis acid activating effect of a THF solvated lithium cation in an organolithium addition to an aldehyde. PMID- 18419119 TI - A catalytic enantioselective conjugate addition of cyanide to enones. AB - The first synthetically useful catalytic enantioselective conjugate addition of cyanide to enones is described. The optimized conditions involved a Gd catalyst (5 or 10 mol %) derived from ligands 3 or 4 and a 1:1 ratio of TBSCN and 2,6 dimethylphenol. The reaction exhibited excellent to high enantioselectivity and a wide substrate scope. Moreover, the 1,4-adduct was exclusively produced over the 1,2-adduct. The complete regioselectivity was due both to stabilization of the 1,4-selective silylated polymetallic catalyst (7) using a TBS group and the ability of the asymmetric catalyst to promote retro-cyanation from the 1,2 adduct. PMID- 18419120 TI - A bridging hexazene (RNNNNNNR) ligand from reductive coupling of azides. AB - This communication reports the first examples of transition metal complexes containing an RNNNNNNR 2- ligand. Addition of 1-azidoadamantane to the diiron(I) synthon LRFeNNFeL R (L R = HC[C(R)N(2,6- iPr 2C 6H 3)] 2; R = methyl, tert-butyl) leads to the diiron complexes L RFe(mu-eta2:eta2-AdN6Ad)FeLR, which are surprisingly thermally stable. Magnetic, Mossbauer, and crystallographic data are consistent with pairs of high-spin iron(II) ions antiferromagnetically coupled through a dianionic AdN6Ad 2- bridge. PMID- 18419121 TI - Synthesis of diamond-shape titanate molecular sheets with different sizes and realization of quantum confinement effect during dimensionality reduction from two to zero. AB - Synthesis of semiconductor nanoparticles with uniform shapes, sizes, and compositions in series with a gradual size reduction has not been achieved for two-dimensional molecular sheets. We report a large-scale (>2.6 g) synthesis of 0.75-nm-thick diamond-shape lepidocrocite-type titanate molecular sheets with the sizes decreasing from (27.3, 19.1) to (7.7, 5.5), where the numbers in parentheses represent the long and short diagonal lengths, respectively, in nm. This is the first example of synthesizing semiconductor nanoparticles in series with the dimensionality reduction from two to zero, without coating the surfaces with surface-passivating ligands. The titanate molecular sheets showed three exciton-absorption bands in the 4.0-6.5 eV region, the absorption energies of which increased with decreasing the area. Contrary to the common belief, the per unit cell oscillator strengths gradually increased with increasing area and the per-particle oscillator strengths increased in proportion to the area. The average reduced exciton masses along the two diagonal axes were 0.10 and 0.11 m e, respectively, which were much smaller than those of bulk titanates (by 60-130 times). The estimated average Bohr radii along the two-diagonal axes were 4.8 and 4.3 nm, respectively. PMID- 18419122 TI - Inherent chirality dominates the visible/near-ultraviolet CD spectrum of rhodopsin. AB - The visible (alpha) and near-UV (beta) CD bands of rhodopsin have been studied extensively, but their source(s) have never been definitively established. Do they result from the intrinsic chirality of the polyene chromophore of the protonated Schiff base of retinal (retPSB) or from the coupling of the transitions of this chromophore with those of protein groups? We have calculated the contributions of these two mechanisms to the CD of rhodopsin. The intrinsic CD of the retPSB chromophore was calculated using time-dependent density functional theory (TDDFT) and, for comparison, the semiempirical ZINDO method. First-order perturbation theory was used to calculate the effects of coupling of the retPSB transitions with the pi pi* transitions of the aromatic chromophores and the pi pi* and n pi* transitions of the peptide groups in rhodopsin. Calculations were performed for eight structures based upon the two molecules in the asymmetric unit of four crystal structures. The most reliable results were obtained from TDDFT calculations on the structure of Okada et al. (J. Mol. Biol. 2004, 342, 571), PDB 1U19. Averaging over the two molecules in the asymmetric unit, the intrinsic rotational strengths are 0.62 +/- 0.00 DBM (Debye-Bohr magneton) and 0.90 +/- 0.03 DBM for the alpha- and beta-bands, respectively. The contributions from coupling with protein groups are, respectively, -0.32 +/- 0.05 and -0.01 +/- 0.03 DBM. Our results show that the visible/near-UV CD bands of rhodopsin are determined by the intrinsic chirality of the retPSB chromophore and that the contributions of coupling with the protein are significantly smaller for the alpha-band and negligible for the beta-band. PMID- 18419123 TI - Differential dynamical effects of macromolecular crowding on an intrinsically disordered protein and a globular protein: implications for in-cell NMR spectroscopy. AB - In-cell NMR provides a valuable means to assess how macromolecules, with concentrations up to 300 g/L in the cytoplasm, affect the structure and dynamics of proteins at atomic resolution. Here an intrinsically disordered protein, alpha synuclein (alphaSN), and a globular protein, chymotrypsin inhibitor 2 (CI2) were examined by using in-cell NMR. High-resolution in-cell spectra of alphaSN can be obtained, but CI2 leaks from the cell and the remaining intracellular CI2 is not detectable. Even after stabilizing the cells from leakage by using alginate encapsulation, no CI2 signal is detected. From in vitro studies we conclude that this difference in detectability is the result of the differential dynamical response of disordered and ordered proteins to the changes of motion caused by the increased viscosity in cells. PMID- 18419124 TI - Molecular engineering of organic sensitizers for dye-sensitized solar cell applications. AB - Novel unsymmetrical organic sensitizers comprising donor, electron-conducting, and anchoring groups were engineered at a molecular level and synthesized for sensitization of mesoscopic titanium dioxide injection solar cells. The unsymmetrical organic sensitizers 3-(5-(4-(diphenylamino)styryl)thiophen-2-yl)-2 cyanoacrylic acid (D5), 3-(5-bis(4-(diphenylamino)styryl)thiophen-2-yl)-2 cyanoacrylic acid (D7), 5-(4-(bis(4-methoxyphenylamino)styryl)thiophen-2-yl)-2 cyanoacrylic acid (D9), and 3-(5-bis(4,4'-dimethoxydiphenylamino)styryl)thiophen 2-yl)-2-cyanoacrylic acid (D11) anchored onto TiO2 and were tested in dye sensitized solar cell with a volatile electrolyte. The monochromatic incident photon-to-current conversion efficiency of these sensitizers is above 80%, and D11-sensitized solar cells yield a short-circuit photocurrent density of 13.90 +/ 0.2 mA/cm(2), an open-circuit voltage of 740 +/- 10 mV, and a fill factor of 0.70 +/- 0.02, corresponding to an overall conversion efficiency of 7.20% under standard AM 1.5 sun light. Detailed investigations of these sensitizers reveal that the long electron lifetime is responsible for differences in observed open circuit potential of the cell. As an alternative to liquid electrolyte cells, a solid-state organic hole transporter is used in combination with the D9 sensitizer, which exhibited an efficiency of 3.25%. Density functional theory/time-dependent density functional theory calculations have been employed to gain insight into the electronic structure and excited states of the investigated species. PMID- 18419125 TI - Optimization and validation of analytical conditions for cholesterol and cholesterol oxides extraction in chicken meat using response surface methodology. AB - The analytical conditions for the extraction of cholesterol and cholesterol oxides in chicken meat were optimized by means of response surface methodology. The separation and identification were performed by normal phase HPLC using UV and refractive index (RI) detectors, and the confirmation of the 11 cholesterol oxides identities in the samples was verified by HPLC-APCI-MS. The developed methodology showed good analytical performance, presenting recovery levels from 84 to 103% and detection limits varying from 0.01 to 0.06 microg/g for UV detection and from 1.98 to 2.12 microg/g for RI detection. The present study demonstrated the presence of 22 R-hydroxycholesterol, 24 S-hydroxycholesterol, and 22 S-hydroxycholesterol for the first time in chicken meat. PMID- 18419126 TI - Analysis of the peptide profile of milk and its changes during thermal treatment and storage. AB - In this study a new method was developed for analysis of the low molecular weight protein fraction of milk, allowing a simple and fast overview of the peptide profile of various milk samples. For this purpose, immobilized metal affinity chromatography (IMAC) was coupled with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). By this technique, two major peptides in milk could be identified as fragments of alpha s1-casein. During heat treatment of raw milk, five new peptides were generated, the origin of which could be assigned to the casein fraction. Storage experiments with extended shelf life milk at 4 degrees C did not show any changes in the peptide profile, whereas in ultra high temperature milk stored at room temperature, one peptide increased significantly, which was identified as the N terminus of alpha-s1-casein. The peptide was assumed to be formed in an enzymatic reaction, which was confirmed in a storage experiment with sterilized milk. Analyses of different commercially available milk samples confirmed the results obtained with the heated and stored milk. Furthermore, differences in the peptide profiles of the samples, probably due to different cow breeds or lactation stages, were observed. These results establish IMAC prior to MALDI-TOF-MS as a valid tool for the rapid analysis of the peptide profile of milk. PMID- 18419127 TI - Sampling and modeling for the quantification of adventitious genetically modified presence in maize. AB - The coexistence of genetically modified (GM) and non-GM crops is an important economic and political issue in the European Union. We examined the GM content in non-GM maize crops in Spain in 2005. Both the standing crop and the harvest were tested, and the %GM DNA was quantified by real-time polymerase chain reaction. We compared the level of GM as a function of distance from known GM source fields in a 1.2 km2 landscape. The distribution of GM was compared to predictions from previous studies, and good agreement was found. Control and monitoring of adventitious GM presence in non-GM crops can only be achieved by fit-for-purpose sampling and testing schemes. We used a GM dispersal function to simulate non-GM crops in the studied zone and tested the accuracy of five different sampling schemes. Random sampling was found to be the most accurate and least susceptible to bias by GM spatial structure or gradients. Simulations showed that to achieve greater than 95% confidence in a GM labeling decision of a harvest (when treated as a single marketed lot), 34 samples would be needed when the harvest was outside 50% of the GM threshold value. The number of samples required increased rapidly as the harvest approached the GM threshold, implying that accurate labeling when the harvest is within +/-17% of the threshold may not be possible with high confidence. PMID- 18419128 TI - Chemical composition of vintage preban absinthe with special reference to thujone, fenchone, pinocamphone, methanol, copper, and antimony concentrations. AB - Thirteen samples of authentic absinthe dating from the preban era (i.e., prior to 1915) were analyzed for parameters that were hypothesized as contributing to the toxicity of the spirit, including naturally occurring herbal essences (thujone, pinocamphone, fenchone), methanol, higher alcohols, copper, and antimony. The total thujone content of preban absinthe was found to range between 0.5 and 48.3 mg/L, with an average concentration of 25.4 +/- 20.3 mg/L and a median concentration of 33.3 mg/L. The authors conclude that the thujone concentration of preban absinthe was generally overestimated in the past. The analysis of postban (1915-1988) and modern commercial absinthes (2003-2006) showed that the encompassed thujone ranges of all absinthes are quite similar, disproving the supposition that a fundamental difference exists between preban and modern absinthes manufactured according to historical recipes. Analyses of pinocamphone, fenchone, base spirits, copper, and antimony were inconspicuous. All things considered, nothing besides ethanol was found in the absinthes that was able to explain the syndrome "absinthism". PMID- 18419129 TI - Oak ellagitannins suppress the phosphorylation of the epidermal growth factor receptor in human colon carcinoma cells. AB - The ellagitannins castalagin and vescalagin, and the C-glycosides grandinin and roburin E as well as ellagic acid were found to potently inhibit the growth of human colon carcinoma cells (HT29) in vitro. In a cell-free system these compounds were identified as potent inhibitors of the protein tyrosine kinase activity of the epidermal growth factor receptor (EGFR) with IC 50 values in the low nanomolar range. To address the question of whether the interference with the activity of the isolated EGFR also plays a role within intact cells, effects on the phosphorylation status of the EGFR, as a measure for its activity, were determined in HT29 cells. As exemplified for castalagin and grandinin, both the nonglycosylated and the glycosylated ellagitannins effectively suppressed EGFR phosphorylation, but only at concentrations > or =10 microM, thus, in a concentration range where growth inhibition was observed. These results indicate that the suppression of EGFR-mediated signaling might contribute to the growth inhibitory effects of these compounds present in oak-matured wines and spirits such as whiskey. In contrast, despite substantial growth inhibitory properties, ellagic acid did not significantly affect EGFR phosphorylation in HT29 cells up to 100 microM. PMID- 18419130 TI - Mapping the membrane topology and extracellular ligand binding domains of the retinol binding protein receptor. AB - STRA6 is a multitransmembrane domain protein not homologous to any other proteins with known function. It functions as the high-affinity receptor for plasma retinol binding protein (RBP) and mediates cellular uptake of vitamin A from the vitamin A-RBP complex. Consistent with the diverse roles of vitamin A and the wide tissue expression pattern of STRA6, mutations in STRA6 are associated with severe pathological phenotypes in humans. The structural basis for STRA6's biochemical function is unknown. Although computer programs predict 11 transmembrane domains for STRA6, its topology has never been studied experimentally. Elucidating the transmembrane topology of STRA6 is critical for understanding its structure and function. By inserting an epitope tag into all possible extracellular and intracellular domains of STRA6, we systematically analyzed the accessibility of each tag on the surface of live cells, the accessibility of each tag in permeabilized cells, and the effect of each tag on RBP binding and STRA6-mediated vitamin A uptake from the vitamin A-RBP complex. In addition, we used a new lysine accessibility technique combining cell-surface biotinylation and tandem-affinity purification to study a region of the protein not revealed by the epitope tagging method. These studies not only revealed STRA6's extracellular, transmembrane, and intracellular domains but also implicated extracellular regions of STRA6 in RBP binding. PMID- 18419131 TI - Role of the beta4Thr-beta73Asp hydrogen bond in HbS polymer and domain formation from multinucleate-containing clusters. AB - Fiber formation and domain formation from deoxy-HbS as well as from beta4 and beta73 HbS variants were investigated after temperature jump using DIC microscopy to gain a basic understanding of the determinants involved. Oversaturated deoxy HbS generated numerous 14-stranded fibers and formed ovoid-shaped, multispherulitic domains. Domain number increased linearly as a function of time. Oversaturated deoxy-alpha2beta2(E6V,T4S) also generated time-dependent, ovoid shaped spherulitic domains like HbS and alpha 2beta2(E6V,D73H) in the deoxy form. In contrast, alpha 2beta2(E6V,T4Y) and HbC-Harlem (alpha2beta2(E6V,D73N)) in the deoxy form generated time-dependent, ball-shaped domains containing many straight, crystalline-like fibers without evidence of branching. Some of these domains formed large needlelike crystals after overnight incubation. The inhibitory effect on polymer formation by beta4Tyr in HbS was stronger than that by beta4Ser but weaker than that by beta73Asn or beta73Leu. In contrast, both deoxy- and oxy-alpha2beta2(E6V,T4V) promoted formation of tiny, disordered amorphous aggregates without a delay time like oxy-HbS, which is in contrast to formation after a delay time of needlelike fibers for alpha 2beta2(E6V,D73L). Solubilities for both deoxy- and oxy-alpha 2beta2(E6V,T4V) were similar to that of deoxy-alpha 2beta2(E6V,D73H) but approximately 10-fold lower than that of deoxy-HbS. These results suggest that the strength of the hydrogen bond between beta4Thr and beta73Asp and the balance between the hydrogen bond and beta6Val hydrophobic interactions in deoxy-HbS polymers control formation of different types of fibers in a single domain or lead to formation of disordered, non nucleated amorphous aggregates. These results also lead to a model in which multinucleation rather than a single-nucleation event occurs in a single cluster to generate numerous fibers growing from a single domain. PMID- 18419132 TI - Thermal stability of the K+ channel tetramer: cation interactions and the conserved threonine residue at the innermost site (S4) of the KcsA selectivity filter. AB - The selectivity filter of most K+ channels contains a highly conserved Thr residue that uniquely forms the S4 binding site for K+ by dual coordination with the backbone carbonyl oxygen and side chain hydroxyl of the same residue. This study examines the effect of mutations of Thr75 in the S4 site of theKcsA K+ channel on the cation dependence of the thermal stability of the tetramer, a phenomenon that reflects the structural role of cations in the filter. Conservative mutations of Thr75 destabilize the tetramer and alter its temperature dependence. Replacement of Thr with Ala or Cys lowers the apparent affinity ofK+, Rb+, and Cs+ for tetramer stabilization by factors ranging from 4- to 14-fold. These same mutations lower the apparent affinity of Ba2+ by approximately 10(3)- or approximately 10(4)-fold for Ala and Cys substitution, respectively,consistent with the known preference of the S4 site for Ba2+. In contrast, substitution of Ala or Cys at T75 anomalously enhances the ability of Na+ to stabilize the tetramer, suggesting that the native Thr residue at S4 is important for ultrahigh K+/Na+ selectivity of K+ channel pores. Elevated temperature orCu2+ cation catalyzes formation of covalent dimers of the T75C mutant of KcsA via formation of disulfide bonds between Cys residues of adjacent subunits. Thiophilic cations such as Hg2+ and Ag+ specifically protect the T75C tetramer against heat-induced dimer formation, demonstrating the contribution of cation interactions to tetramer stability in a channel with a non-native S4 site engineered to bind foreign cations. PMID- 18419133 TI - Unnatural amino acid replacement in a yeast G protein-coupled receptor in its native environment. AB - Ste2p is the G protein-coupled receptor (GPCR) for the tridecapeptide pheromone alpha factor of Saccharomyces cerevisiae. This receptor-pheromone pair has been used extensively as a paradigm for investigating GPCR structure and function. Expression in yeast harboring a cognate tRNA/aminoacyl-tRNA synthetase pair specifically evolved to incorporate p-benzoyl- l-phenylalanine (Bpa) in response to the amber codon allowed the biosynthesis of Bpa-substituted Ste2p in its native cell. We replaced natural amino acid residues in Ste2p with Bpa by engineering amber TAG stop codons into STE2 encoded on a plasmid. Several of the expressed Bpa-substituted Ste2p receptors exhibited high-affinity ligand binding, and incorporation of Bpa into Ste2p influenced biological activity as measured by growth arrest of whole cells in response to alpha factor. We found that, at concentrations of 0.1-0.5 mM, a dipeptide containing Bpa could be used to enhance delivery of Bpa into the cell, while at 2 mM, both dipeptide and Bpa were equally effective. The application of a peptide delivery system for unnatural amino acids will extend the use of the unnatural amino acid replacement methodology to amino acids that are impermeable to yeast. Incorporation of Bpa into Ste2p was verified by mass spectrometric analysis, and two Bpa-Ste2p mutants were able to selectively capture alpha factor into the ligand-binding site after photoactivation. To our knowledge, this is the first experimental evidence documenting an unnatural amino acid replacement in a GPCR expressed in its native environment and the use of a mutated receptor to photocapture a peptide ligand. PMID- 18419134 TI - Regulatory control of human cytosolic branched-chain aminotransferase by oxidation and S-glutathionylation and its interactions with redox sensitive neuronal proteins. AB - Redox regulation of proteins through oxidation and S-thiolation are important regulatory processes, acting in both a protective and adaptive role in the cell. In the current study, we investigated the sensitivity of the neuronal human cytosolic branched-chain aminotransferase (hBCATc) protein to oxidation and S thiolation, with particular attention focused on functionality and modulation of its CXXC motif. Thiol specific reagents showed significant redox cycling between the reactive thiols and the TNB anion, and using NEM, four of the six reactive thiols are critical to the functionality of hBCATc. Site-directed mutagenesis studies supported these findings where a reduced kcat (ranging from 50-70% of hBCATc) for C335S, C338S, C335/8S, and C221S, respectively, followed by a modest effect on C242S was observed. However, only the thiols of the CXXC motif (C335 and C338) were directly involved in the reversible redox regulation of hBCATc through oxidation (with a loss of 40-45% BCAT activity on air oxidation alone). Concurrent with these findings, under air oxidation, the X-ray crystallography structure of hBCATc showed a disulphide bond between C335 and C338. Further oxidation of the other four thiols was not evident until levels of hydrogen peroxide were elevated. S-thiolation experiments of hBCATc exposed to GSH provided evidence for significant recycling between GSH and the thiols of hBCATc, which implied that under reducing conditions GSH was operating as a thiol donor with minimal S-glutathionylation. Western blot analysis of WT hBCATc and mutant proteins showed that as the ratio of GSH:GSSG decreased significant S glutathionylation occurred (with a further loss of 20% BCAT activity), preferentially at the thiols of the CXXC motif, suggesting a shift in function toward a more protective role for GSH. Furthermore, the extent of S glutathionylation increased in response to oxidative stress induced by hydrogen peroxide potentially through a C335 sulfenic acid intermediate. Deglutathionylation of hBCATc-SSG using the GSH/glutaredoxin system provides evidence that this protein may play an important role in cellular redox regulation. Moreover, redox associations between hBCATc and several neuronal proteins were identified using targeted proteomics. Thus, our data provides strong evidence that the reactive thiol groups, in particular the thiols of the CXXC motif, play an integral role in redox regulation and that hBCATc has redox mediated associations with several neuronal proteins involved in G-protein cell signaling, indicating a novel role for hBCATc in cellular redox control. PMID- 18419135 TI - Interaction of a peptide derived from glycoprotein gp36 of feline immunodeficiency virus and its lipoylated analogue with phospholipid membranes. AB - P59, a 20-mer peptide modeled on the membrane-proximal external region (MPER) of the feline immunodeficiency virus (FIV) gp36 ectodomain, has potent antiviral activity. The lipoylated analogue, lipo-P59, displays a similar activity, which is preferentially retained by cellular substrates. A mechanism has been proposed recently in which the peptide, being positioned on the surface of the cell membrane, inhibits its fusion with the virus; the lipophilic chain of lipo-P59 is thought to insert into the membrane interior, thus anchoring the peptide at the surface. In the present work, lipid-peptide interactions of P59 and lipo-P59 with phospholipid liposomes are investigated using spin-label electron spin resonance spectroscopy. Two phospholipids have been examined, the zwitterionic dimyristoyl phosphatidylcholine and the anionic dimyristoyl phosphatidylglycerol, and a wide range of lipid spin labels, including positional isomers. Independent of the membrane charge, both peptides bind to lipid bilayers; however, whereas P59 insertion between the lipid headgroups leads to significant liposome destabilization, eventually resulting in vesicle fragmentation with the formation of smaller aggregates, lipo-P59 inserts with the lipophilic tail among the lipid chains, while the peptidic portion remains adsorbed onto the membrane, where it can effectively exert its antiviral activity. PMID- 18419136 TI - Chemo-enzymatic detection of protein isoaspartate using protein isoaspartate methyltransferase and hydrazine trapping. AB - Isoaspartate formation is a ubiquitous post-translation modification arising from spontaneous asparagine deamidation or aspartate isomerization. The formation of isoaspartate inserts a methylene group into the protein backbone, generating a "kink", and may drastically alter protein structure and function, thereby playing critical roles in a myriad of biological processes, human diseases, and protein pharmaceutical development. Herein, we report a chemo-enzymatic detection method for the isoaspartate protein, which in particular allows the affinity enrichment of isoaspartate-containing proteins. In the initial step, protein isoaspartate methyltransferase selectively converts isoaspartates into the corresponding methyl esters. Subsequently, the labile methyl ester is trapped by strong nucleophiles in aqueous solutions, such as hydrazines to form hydrazides. The stable hydrazide products can be analyzed by standard proteomic techniques, such as matrix-assisted laser desorption ionization and electrospray ionization mass spectrometry. Furthermore, the chemical trapping step allows us to introduce several tagging strategies for product identification and quantification, such as UV-vis and fluorescence detection through a dansyl derivative. Most significantly, the hydrazide product can be enriched by affinity chromatography using aldehyde resins, thus drastically reducing sample complexity. Our method hence represents the first technique for the affinity enrichment of isoaspartyl proteins and should be amendable to the systematic and comprehensive characterization of isoaspartate, particularly in complex systems. PMID- 18419137 TI - Selective extraction and enrichment of multiphosphorylated peptides using polyarginine-coated diamond nanoparticles. AB - Despite recent advances in phosphopeptide research, detection and characterization of multiply phosphorylated peptides have been a challenge. This work presents a new strategy that not only can effectively extract phosphorylated peptides from complex samples but also can selectively enrich multiphosphorylated peptides for direct matrix-assisted laser desorption/ionization time-of-flight mass spectrometric analysis. Polyarginine-coated diamond nanoparticles are the solid-phase extraction supports used for this purpose. The supports show an exceptionally high affinity for multiphosphorylated peptides due to multiple arginine-phosphate interactions. The efficacy of this method was demonstrated by analyzing a small volume (50 microL) of tryptic digests of proteins such as beta casein, alpha-casein, and nonfat milk at a concentration as low as 1 x 10 (-9) M. The concentration is markedly lower than that can be achieved by using other currently available technologies. We quantified the enhanced selectivity and detection sensitivity of the method using mixtures composed of mono- and tetraphosphorylated peptide standards. This new affinity-based protocol is expected to find useful applications in characterizing multiple phosphorylation sites on proteins of interest in complex and dilute analytes. PMID- 18419139 TI - NMR-based characterization of metabolic alterations in hypertension using an adaptive, intelligent binning algorithm. AB - As with every -omics technology, metabolomics requires new methodologies for data processing. Due to the large spectral size, a standard approach in NMR-based metabolomics implies the division of spectra into equally sized bins, thereby simplifying subsequent data analysis. Yet, disadvantages are the loss of information and the occurrence of artifacts caused by peak shifts. Here, a new binning algorithm, Adaptive Intelligent Binning (AI-Binning), which largely circumvents these problems, is presented. AI-Binning recursively identifies bin edges in existing bins, requires only minimal user input, and avoids the use of arbitrary parameters or reference spectra. The performance of AI-Binning is demonstrated using serum spectra from 40 hypertensive and 40 matched normotensive subjects from the Asklepios study. Hypertension is a major cardiovascular risk factor characterized by a complex biochemistry and, in most cases, an unknown origin. The binning algorithm resulted in an improved classification of hypertensive status compared with that of standard binning and facilitated the identification of relevant metabolites. Moreover, since the occurrence of noise variables is largely avoided, AI-Binned spectra can be unit-variance scaled. This enables the detection of relevant, low-intensity metabolites. These results demonstrate the power of AI-Binning and suggest the involvement of alpha-1 acid glycoproteins and choline biochemistry in hypertension. PMID- 18419138 TI - Elastomeric microchip electrospray emitter for stable cone-jet mode operation in the nanoflow regime. AB - Despite widespread interest in combining laboratory-on-a-chip technologies with mass spectrometry (MS)-based analyses, the coupling of microfluidics to electrospray ionization (ESI)-MS remains challenging. We report a robust, integrated poly(dimethylsiloxane) microchip interface for ESI-MS using simple and widely accessible microfabrication procedures. The interface uses an auxiliary channel to provide electrical contact for the stable cone-jet electrospray without sample loss or dilution. The electric field at the channel terminus is enhanced by two vertical cuts that cause the interface to taper to a line rather than to a point, and the formation of a small Taylor cone at the channel exit ensures subnanoliter postcolumn dead volumes. Cone-jet mode electrospray was demonstrated for up to 90% aqueous solutions and for extended durations. Comparable ESI-MS sensitivities were achieved using both microchip and conventional fused silica capillary emitters, but stable cone-jet mode electrosprays could be established over a far broader range of flow rates (from 50-1000 nL/min) and applied potentials using the microchip emitters. This attribute of the microchip emitter should simplify electrospray optimization and make the stable electrospray more resistant to external perturbations. PMID- 18419140 TI - Synthesis, microsome-mediated metabolism, and identification of major metabolites of environmental pollutant naphtho[8,1,2-ghi]chrysene. AB - Naphtho[8,1,2- ghi]chrysene, commonly known as naphtho[1,2- e]pyrene (N[1,2- e]P) is a widespread environmental pollutant, identified in coal tar extract, air borne particulate matter, marine sediment, cigarette smoke condensate, and vehicle exhaust. Herein, we determined the ability of rat liver microsomes to metabolize N[1,2- e]P and an unequivocal assignment of the metabolites by comparing them with independently synthesized standards. We developed the synthesis of both the fjord region and the K-region dihydrodiols and various phenolic derivatives for metabolite identification. The 12-OH-N[1,2- e]P, fjord region dihydrodiol 14 and diol epoxide 15 were synthesized using a Suzuki cross coupling reaction followed by the appropriate manipulation of the functional groups. The K-region trans-4,5-dihydrodiol ( 18) was prepared by the treatment of N[1,2- e]P with OsO 4 to give cis-dihydrodiol 16, followed by pyridinium chlorochromate oxidation to quinone 17, and finally reduction with NaBH 4 to afford the dihydrodiol 18 with the desired trans stereochemistry. The 9-OH-N[1,2- e]P ( 30) and N[1,2- e]P trans-9,10-dihydrodiol ( 32) were also synthesized following a Suzuki cross-coupling approach starting from 1,2,3,6,7,8 hexahydropyrene-4-boronic acid. The metabolism of N[1,2- e]P with rat liver microsomes led to several dihydrodiol and phenolic metabolites as assessed by the HPLC trace. The 11,12-dihydrodiol and 4,5-dihydrodiol were identified as major dihydrodiol metabolites. The synthesized 9,10-dihydrodiol, on the other hand, did not match with any of the peaks in the metabolism trace. Among the phenols, only 12-OH-N[1,2- e]P was identified in the metabolism. The other phenolic derivatives synthesized, that is, the 4-/5-, 9-, 10-, and 11-hydroxy derivatives, were not detected in the metabolism trace. In summary, N[1,2- e]P trans-11,12-dihydrodiol was the major metabolite formed along with N[1,2- e]P 4,5- trans-dihydrodiol and 12-OH-N[1,2- e]P on exposure of rat liver microsomes to N[1,2- e]P. The presence of N[1,2- e]P in the environment and formation of fjord region dihydrodiol 14 as a major metabolite in in vitro metabolism studies strongly suggest the role of N[1,2- e]P as a potential health hazard. PMID- 18419141 TI - Bioactivation of diclofenac in vitro and in vivo: correlation to electrochemical studies. AB - Diclofenac is widely used in the treatment of, for example, arthritis and muscle pain. The use of diclofenac has been associated with hepatotoxicity, which has been linked to the formation of reactive metabolites. Diclofenac can be metabolized to 4'-OH- and 5-OH-diclofenac, both of which are able to form quinone imines capable of reacting with, for example, GSH and nucleophilic groups in proteins. Electrochemistry has been shown to be a suitable tool for mimicking some types of oxidative drug metabolism and for studying the formation of reactive metabolites. In these studies, the electrochemical oxidation of diclofenac to a +16 Da metabolite was shown to be identical to a synthetic standard of 5-OH-diclofenac. Furthermore, two different experimental designs were investigated with respect to the electrochemical oxidation of 4'-OH- and 5-OH diclofenac. In the first approach, the oxidized sample was collected in an aqueous solution of GSH, whereas in the other approach, GSH was added to the sample before the oxidation was performed. From these electrochemical oxidations, a range of GSH conjugates of 4'-OH- and 5-OH-diclofenac were observed and characterized by MS/MS. This allowed the development of sensitive LC-MS methods in order to detect the GSH conjugates from in vivo (rat bile) and in vitro (human liver microsomes (HLM), rat liver microsomes (RLM), and rat hepatocytes) samples. A wide range of mono-, di-, and triglutathionyl conjugates were detected in the in vitro and in vivo samples. It was also observed that 5-OH-diclofenac formed GSH conjugates with RLM and HLM without addition of NADPH, whereas GSH conjugate formation of 4'-OH-diclofenac was NADPH-dependent. This indicated that 5-OH diclofenac was prone to auto-oxidation. The oxidation potentials of the two hydroxy metabolites were determined by cyclic voltammetry. A difference of 69 mV was observed between the two oxidation potentials, which in part may explain the extent of auto-oxidation for 5-OH-diclofenac. In conclusion, it was shown that electrochemical oxidation was capable of mimicking the metabolic hydroxylation of diclofenac to 5-OH-diclofenac. Furthermore, electrochemical oxidation was used to generate a range of GSH conjugates of 4'-OH- and 5-OH-diclofenac and a number of these conjugates were also detected in metabolism studies with microsomes (HLM/RLM) and freshly isolated rat hepatocytes, and in vivo in rat bile. PMID- 18419142 TI - Microwave-assisted organic synthesis and transformations using benign reaction media. AB - A nonclassical heating technique using microwaves, termed "Bunsen burner of the 21st century", is rapidly becoming popular and is dramatically reducing reaction times. The significant outcomes of microwave (MW)-assisted green chemistry endeavors, which have resulted in the development of synthetic protocols for drugs and fine chemicals synthesis that are relatively more sustainable, are summarized. The use of emerging microwave-assisted chemistry techniques in conjunction with greener reaction media is dramatically reducing chemical waste and reaction times in several organic syntheses and chemical transformations. A brief historic account of our own experiences in developing MW-assisted organic transformations, which involve various benign alternatives, such as solid supported reagents, and greener reaction media, namely, aqueous, ionic liquid, and solvent-free, for the synthesis of various heterocycles, coupling reactions, oxidation-reduction reactions, and some name reactions are described. Synthesis of Heterocycles. The synthetic chemistry community has been under increased pressure to produce, in an environmentally benign fashion, the myriad of heterocyclic systems required by society in a short span of time, and one of the best options to accelerate these synthetic processes is to use MW technology. The efficient use of the MW heating approach for the synthesis of various heterocyclic compounds in aqueous and solvent-free medium is discussed. Organic Named Reactions. The application of MW chemistry for various named reaction such as the Prins reaction, the Suzuki reaction, the Heck reaction, the Aza-Michael reaction, Trost's gamma-addition, and the Cannizzaro reaction are summarized. Synthesis and Application of Ionic Liquids. Ionic liquids (ILs), being polar and ionic, in character couple with MW irradiation very efficiently and are, therefore, ideal MW-absorbing candidates for expediting chemical reactions. MW assisted solvent-free synthesis and application of ILs are discussed. Oxidation Reduction Reactions. MW protocols using mineral oxides such as alumina, silica, and clay to immobilize reagents on such solid supports have been extensively explored under "dry" media conditions. Various solvent-free examples of oxidation reactions are discussed that involve mixing of neat substrates with clay supported iron(III) nitrate (clayfen) or iodobenzene diacetate (IBD) as an oxidant; some interesting MW reduction protocols using borohydrides are also discussed. Protection-Deprotection Reactions. The protection and deprotection of alcohols and amines are common events in multistep organic syntheses. Various protection and deprotection protocols under MW irradiation are discussed, including tetrahydropyranylation and (benzyloxycarbonyl) (Cbz)-protection, which are the most frequently employed methods. PMID- 18419143 TI - Characterization of the growth of polyelectrolyte multilayers formed at interfaces between aqueous phases and thermotropic liquid crystals. AB - Polyelectrolyte multilayers (PEMs) formed at interfaces between aqueous solutions and thermotropic (water-immiscible) liquid crystals (LCs) offer the basis of a new method to tailor the nanometer-scale structure and chemical functionality of these interfaces. Toward this end, we report a study that compares the growth of PEMs formed at mobile and deformable interfaces defined by LCs relative to growth observed at model (rigid) solid surfaces. Experiments aimed at determining if polyelectrolytes such as poly(sodium-4-styrenesulfonate) (PSS) can partition from the aqueous phase into the bulk of the LC yielded no evidence of such partitioning. Whereas measurements of the growth of PEMs formed from poly(allylamine hydrochloride) (PAH) and PSS at the aqueous-LC interface revealed growth characteristics similar to those measured at both hydrophobic and hydrophilic interfaces of solids, the growth of PEMs from PAH and poly(acrylic acid) (PAA) at the aqueous-LC interface was found to differ substantially from the solids investigated: (i) the linear growth of PEMs of PAH/PAA that was measured at the aqueous-LC interface under conditions that did not lead to the growth of PEMs at the interface of octadecyltrichlorosilane (OTS)-treated glass (a hydrophobic solid surface), and (ii) in comparison to the growth of PEMs of PAH/PAA at the surface of glass (a hydrophilic charged surface), a higher rate of growth was observed at the aqueous-LC interface. The finding that the growth rate of PEMs of PAH/PAA at aqueous-LC interfaces is greater than on solid surfaces is supported by additional measurements of growth as a function of pH. Finally, the pH-triggered reorganization of PAH/PAA PEMs supported at the aqueous-LC interface led to changes in the order and optical properties of the LC. These data are discussed in light of the nature of aqueous-LC interfaces, including the mobility and deformability of the interface and recent measurements of the zeta-potentials of aqueous-LC interfaces. PMID- 18419144 TI - Nanoparticle stability in semidilute and concentrated polymer solutions. AB - The wetting of PDMS-grafted silica spheres (PDMS- g-silica) is connected to their depletion restabilization in semidilute and concentrated PDMS/cyohexane polymer solutions. Specifically, we found that a wetting diagram of chemically identical graft and free homopolymers predicts stability of hard, semisoft, and soft spheres as a function of the bulk free polymer volume fraction, graft density, and the graft and free polymer chain lengths. The transition between stable and aggregated regions is determined optically and with dynamic light scattering. The point of demarcation between the regions occurs when the graft and free polymer chains are equal in length. When graft chains are longer than free chains, the particles are stable; in contrast, the particles are unstable when the opposite is true. The regions of particle stability and instability are corroborated with theoretical self-consistent mean-field calculations, which not only show that the grafted brush is responsible for particle dispersion in the complete wetting region but also aggregation in the incomplete wetting region. Ultimately, our results indicate that depletion restabilization depends on the interfacial properties of the nanoparticles in semidilute and concentrated polymer solutions. PMID- 18419145 TI - Altered phase model for polymer clay nanocomposites. AB - This paper describes a multiscale approach used to model polymer clay nanocomposites (PCNs) based on a new altered phase concept. Constant-force steered molecular dynamics (SMD) is used to evaluate nanomechanical properties of the constituents of intercalated clay units in PCNs, which were used in the finite element model. Atomic force microscopy and nanoindentation techniques provided additional input to the finite element method (FEM) model. FEM is used to construct a representative PCN model that simulates the composite response of intercalated clay units and the surrounding polymer matrix. From our simulations we conclude that, in order to accurately predict mechanical response of PCNs, it is necessary to take into account the molecular-level interactions between constituents of PCN, which are responsible for the enhanced nanomechanical properties of PCNs. This conclusion is supported by our previous finding that there is a change in crystallinity of polymeric phase due to the influence of intercalated clay units. The extent of altered polymeric phase is obtained from observations of a zone of the altered polymeric phase surrounding intercalated clay units in the "phase image" of PCN surface, obtained using an atomic force microscope (AFM). An accurate FEM model of PCN is constructed that incorporates the zone of the altered polymer. This model is used to estimate elastic modulus of the altered polymer. The estimated elastic modulus for the altered polymer is 4 to 5 times greater than that of pure polymer. This study indicates that it is necessary to take into account molecular interactions between constituents in nanocomposites due to the presence of altered phases, and furthermore provides us with a new direction for the modeling and design of nanocomposites. PMID- 18419146 TI - Photoinduced electron transfer in thin layers composed of fullerene-cyclic peptide conjugate and pyrene derivative. AB - A bilayer structure was constructed on gold by Langmuir-Blodgett deposition of a fullerene (C 60)-cyclic peptide-poly(ethylene glycol) (PEG) conjugate and thereafter a pyrene derivative from the air/water interface. The cyclic peptide moiety acts as a scaffold to prevent the fullerenes from self-aggregation and accordingly makes the monolayer homogeneous and stable. In addition to this gold/C 60-cyclic peptide-PEG/pyrene bilayer, a pyrene monolayer, a gold/C 60-PEG conjugate/pyrene bilayer (lacking the peptide scaffold), and a gold/pyrene/C 60 cyclic peptide-PEG bilayer (with the opposite order of layers) were also prepared, and their anodic photocurrent generation were studied in an aqueous solution containing a sacrifice electron donor. The most efficient photocurrent generation was observed in the gold/C 60-cyclic peptide-PEG/pyrene bilayer. It is considered that the C 60 unit acts not only as sensitizer but also as an electron acceptor facilitating the electron transfer from the excited pyrene unit to gold, and that the fullerene layer suppresses quenching of the excited pyrene unit by energy transfer to gold. Furthermore, the cyclic peptide scaffold helps the fullerenes disperse without aggregation in the membrane and seems to protect their redox properties or inhibit self-quenching of their excited state. It is thus concluded that a bilayer structure with desired orientation of functional units is important for efficient photoinduced electron transfer and that a cyclic peptide scaffold is useful to locate hydrophobic functional groups properly in a thin layer. PMID- 18419147 TI - Mechanism of antimicrobial activity of CdTe quantum dots. AB - The antimicrobial activity and mechanism of CdTe quantum dots (QDs) against Escherichia coli were investigated in this report. Colony-forming capability assay and atomic force microscopy (AFM) images show that the QDs can effectively kill the bacteria in a concentration-dependent manner. Results of photoluminescence spectrophotometry, confocal microscopy, and antioxidative response tests indicate that the QDs bind with bacteria and impair the functions of a cell's antioxidative system, including down-regulations of antioxidative genes and decreases of antioxidative enzymes activities. The oxidative damage of protein and lipid is also observed with thiobarbituric reacting substances and protein carbonyl assays, respectively. On the basis of these results, it is proposed that the mechanism of the antimicrobial activity of CdTe QDs involves QDs-bacteria association and a reactive oxygen species-mediated pathway. Thus, CdTe QDs could have the potential to be formulated as a novel antimicrobial material with excellent optical properties. PMID- 18419148 TI - Preparation of pH-sensitive poly(glycidol) derivatives with varying hydrophobicities: their ability to sensitize stable liposomes to pH. AB - We have previously shown that modification with succinylated poly(glycidol) (SucPG) provides stable egg yolk phosphatidylcholine (EYPC) liposomes with pH sensitive fusogenic property. Toward production of efficient pH-sensitive liposomes, in this study, we newly prepared three carboxylated poly(glycidol) derivatives with varying hydrophobicities by reacting poly(glycidol) with glutaric anhydride, 3-methylglutaric anhydride, and 1,2-cyclohexanedicarboxylic anhydride, respectively, designated as GluPG, MGluPG, and CHexPG. Correlation between side-chain structures of these polymers and their respective abilities to sensitize stable liposomes to pH was investigated. These polymers are soluble in water at neutral pH but became water-insoluble in weakly acidic conditions. The pH at which the polymer precipitated was higher in the order SucPG < GluPG < MGluPG < CHexPG, which is consistent with the number of carbon atoms of these polymers' side chains. Although CHexPG destabilized EYPC liposomes even at neutral pH, attachment of other polymers provided pH-sensitive properties to the liposomes. The liposomes bearing polymers with higher hydrophobicity exhibited more intense responses, such as content release and membrane fusion, at mildly acidic pH and achieved more efficient cytoplasmic delivery of membrane impermeable dye molecules. As a result, modification with appropriate hydrophobicity, MGluPG, produced highly potent pH-sensitive liposomes, which might be useful for efficient cytoplasmic delivery of bioactive molecules, such as proteins and genes. PMID- 18419149 TI - Proteomic identification of the Cus system as a major determinant of constitutive Escherichia coli silver resistance of chromosomal origin. AB - Although silver is one of the most potent and rapidly acting toxic metals to bacteria, silver-resistant bacteria do exist with low incidence. A proteomic approach was employed to identify the silver resistance determinants of a silver resistant Escherichia coli strain isolated from stepwise selection against increasing concentrations of silver (Li et al. J. Bacteriol 1997, 179, 6127-32). Two-dimensional gel electrophoresis and mass spectrometry analysis revealed that members of the CusCFBA copper/silver chemiosmotic efflux system were highly expressed in the silver-resistant strain but undetectable in the parental silver sensitive strain. Disruption of the cus locus of the silver-resistant strain resulted in a decrease of the minimum inhibitory concentration of Ag (+) from more than 1 mM to 12 microM. These results suggest that the chromosomally encoded Cus system, which naturally controls the periplasmic copper concentrations, is selectable to confer a constitutive silver resistance phenotype. PMID- 18419150 TI - Hepatocystin is not secreted in cyst fluid of hepatocystin mutant polycystic liver patients. AB - Autosomal dominant polycystic liver disease (PCLD) is characterized by multiple liver cysts and is caused by mutations in PRKCSH (hepatocystin). Mechanisms of cystogenesis are unknown, but previous studies have shown that hepatocystin is secreted in vitro. The goal of this study was to determine the fate of hepatocystin in vivo. Using immunoprecipitation, we determined that mutant hepatocystin is secreted from both apical and basolateral cell surface of MDCK cells stably transfected with mutant hepatocystin. Analysis of 60 cyst fluid samples from polycystic livers using Western blot, MALDI-TOF MS or nLC-MS/MS did not detect hepatocystin in liver cyst fluid. We did identify 163 ubiquitous serum proteins. No paracrine or autocrine factors were recognized. Although cyst fluids vary greatly in protein concentration, a PCLD specific protein pattern was not established. In conclusion, hepatocystin is not secreted in PCLD liver cyst fluid, suggesting that mutant hepatocystin is either not produced or degraded intracellularly. PCLD cysts develop from intralobular bile ductules and cyst fluid mainly contains common serum proteins comparable to that of other polycystic diseases. PMID- 18419151 TI - Isoform analysis of LC-MS/MS data from multidimensional fractionation of the serum proteome. AB - We developed a visualization approach for the identification of protein isoforms, precursor/mature protein combinations, and fragments from LC-MS/MS analysis of multidimensional fractionation of serum and plasma proteins. We also describe a pattern recognition algorithm to automatically detect and flag potentially heterogeneous species of proteins in proteomic experiments that involve extensive fractionation and result in a large number of identified serum or plasma proteins in an experiment. Examples are given of proteins with known isoforms that validate our approach and present a subset of precursor/mature protein pairs that were detected with this approach. Potential applications include identification of differentially expressed isoforms in disease states. PMID- 18419152 TI - Evidence of PPII-like helical conformation and glass transition in a self assembled solid-state polypeptide-surfactant complex: poly(L histidine)/docylbenzenesulfonic acid. AB - We present lamellar self-assembly of cationic poly(L-histidine) (PLH) stoichiometrically complexed with an anionic surfactant, dodecyl benzenesulfonic acid (DBSA), which allows a stabilized conformation reminiscent of polyproline type II (PPII) left-handed helices. Such a conformation has no intrapeptide hydrogen bonds, and it has previously been found to be one source of flexibility, e.g., in collagen and elastin, as well as an intermediate in silk processing. PLH(DBSA)1.0 complexes were characterized by Fourier transform infrared spectroscopy (FTIR), circular dichroism (CD), small-angle X-ray scattering (SAXS), transmission electron microscopy (TEM), and differential scanning calorimetry (DSC). The PPII-like conformation in PLH(DBSA)1.0 is revealed by characteristic CD and FTIR spectra, where the latter indicates absence of intrachain peptide hydrogen bonds. In addition, a glass transition was directly verified by DSC at ca. 135 degrees C for PLH(DBSA)1.0 and indirectly by SAXS and TEM in comparison to pure PLH at 165 degrees C, thus indicating plasticization. Glass transitions have not been observed before in polypeptide-surfactant complexes. The present results show that surfactant binding can be a simple scheme to provide steric crowding to stabilize PPII conformation to tune the polypeptide properties, plasticization and flexibility. PMID- 18419153 TI - Comparison of the adsorbed conformation of barley lipid transfer protein at the decane-water and vacuum-water interface: a molecular dynamics simulation. AB - Molecular dynamics simulation is used to model the adsorption of the barley lipid transfer protein (LTP) at the decane-water and vacuum-water interfaces. Adsorption at both surfaces is driven by displacement of water molecules from the interfacial region. LTP adsorbed at the decane surface exhibits significant changes in its tertiary structure, and penetrates a considerable distance into the decane phase. At the vacuum-water interface LTP shows small conformational changes away from its native structure and does not penetrate into the vacuum space. Modification of the conformational stability of LTP by reduction of its four disulphide bonds leads to an increase in conformational entropy of the molecules, which reduces the driving force for adsorption. Evidence for changes in the secondary structure are also observed for native LTP at the decane-water interface and reduced LTP at the vacuum-water interface. In particular, intermittent formation of short (six-residue) regions of beta-sheet is found in these two systems. Formation of interfacial beta-sheet in adsorbed proteins has been observed experimentally, notably in the globular milk protein beta lactoglobulin and lysozyme. PMID- 18419154 TI - Mono-tetrahydrofuran annonaceous acetogenins from Annona squamosa as cytotoxic agents and calcium ion chelators. AB - Eight new mono-tetrahydrofuran (THF)-type annonaceous acetogenins, squafosacins B, C, F, and G (1-4), squadiolins A-C (5-7), and cis-annotemoyin-1 (8), as well as eight known annonaceous acetogenins, glabranin, annotemoyins-1 and -2, bullatencin, cis-bullatencin, and uvariamicins-I, -II, and -III, were isolated from the seeds of Annona squamosa by HPLC. The structures of all new isolates were elucidated by using spectroscopic and chemical methods. Squadiolins A (5) and B (6) showed ng/mL potency against human Hep G2 hepatoma cells and significant cytotoxic activity against human MDA-MB-231 breast cancer cells. Squafosacin B (1) also exhibited significant cytotoxic activity against human Hep G2 and 3B hepatoma and MCF-7 breast cancer cells. In addition, the chelation of mono-THF acetogenins with calcium ions was investigated using isothermal titration calorimetry. PMID- 18419155 TI - Determination of arylglycerol-beta-aryl ether linkages in enzymatic mild acidolysis lignins (EMAL): comparison of DFRC/(31)P NMR with thioacidolysis. AB - Enzymatic mild acidolysis lignins (EMAL) isolated from different species of softwood and Eucalyptus globulus were submitted to comparative analysis that included thioacidolysis, derivatization followed by reductive cleavage (DFRC), and DFRC followed by quantitative (31)P NMR (DFRC/(31)P NMR). While gas chromatography (GC) was used to determine the monomer yields from both thioacidolysis and DFRC, (31)P NMR studies quantified the various phenolic hydroxy groups released by DFRC. The monomer yields from thioacidolysis and DFRC were substantially different, with thioacidolysis resulting in higher yields. In contrast, an excellent agreement was obtained in the total number of beta-aryl ether structures determined by thioacidolysis and DFRC/(31)P NMR, indicating that the combination of DFRC with quantitative (31)P NMR overcomes, at least in part, the limitations presented by the DFRC method. Both thioacidolysis and DFRC/(31)P NMR were further used to better understand the lignin isolation process from wood. The results show that mild rotary ball milling minimizes, but does not prevent, the degradation of beta-O-4 structures during the early stages of wood pulverization. The extent of such degradation was found to be higher for E. globulus than for a variety of softwoods examined. Furthermore, the structures of the EMALs isolated at yields ranging from 20% to 62% were very similar, indicating structural homogeneity in the lignin biopolymer within the secondary wall. PMID- 18419156 TI - Oleanane-type triterpenes from the flowers and roots of Saussurea muliensis. AB - Six new oleanane-type triterpenes (1- 6), along with five known compounds, were isolated from the flowers and roots of Saussurea muliensis. On the basis of spectroscopic methods, with special emphasis on 1D and 2D NMR techniques, the structures of the new compounds were characterized as 3beta,22alpha dihydroxyolean-12-en-30-oic acid (1), 3alpha-(E)-caffeoyloxyolean-12-en-30-oic acid (2), 3alpha-(E)-coumaroyloxyolean-12-en-30-oic acid (3), 3alpha,22alpha diacetoxy-20beta,21alpha,29-trihydroxy-30-norolean-12-ene (4), 3alpha,22alpha diacetoxy-21alpha,29-dihydroxy-20beta-methoxy-30-norolean-12-ene (5), and 3alpha,22alpha-diacetoxy-20beta,21alpha-dihydroxy-29-palmityloxy-30-norolean-12 ene (6). The isolated compounds (1- 6) were not active against Staphylococcus aureus, Escherichia coli, Bacillus cereus, and Candida albicans. PMID- 18419157 TI - A new cyclization to fused pyrazoles tunable for pericyclic or pseudopericyclic route: an experimental and theoretical study. AB - 2-Pyrazinyl (2) and 3-pyridazinylketone arylhydrazones (6) and their benzologues undergo a ring closure reaction to yield pyrazolo[3,4- b]pyrazines (4) and pyrazolo[4,3- c]pyridazines (7), respectively, in acceptable to good yields. The reaction was found to be accelerated by using acidic or basic conditions. Quantum chemical calculations suggest the key step of the mechanism to be a direct cyclization; analysis of aromaticity based on computed magnetic properties revealed its medium-dependent pericyclic or pseudopericyclic character. The cyclization reaction has also been extended for the synthesis of related ring systems ( 9, 12, 14). PMID- 18419158 TI - Efficient synthesis of 3-iodoindenes via Lewis-acid catalyzed Friedel-Crafts cyclization of iodinated allylic alcohols. AB - A convenient BF(3).Et(2)O-catalyzed Friedel-Crafts cyclization of iodinated allylic alcohols is reported. The present reaction provides an efficient protocol to 3-iodo-1(H)-indene derivatives in good to high yields under mild conditions. Further, the iodoindene derivatives are valuable synthetic building blocks for elaboration of molecular complexity, such as in the construction of multiaryl substituted indenes by Suzuki coupling reaction. PMID- 18419159 TI - A host-induced intramolecular charge-transfer complex and light-driven radical cation formation of a molecular triad with cucurbit[8]uril. AB - The host-guest chemistry of systems containing a molecular triad Ru(bpy) 3-MV (2+)-naphthol complex (denoted as Ru (2+)-MV (2+)-Np, 1) and cucurbit[8]uril (CB[8]) is investigated by NMR, ESI-MS, UV-vis, and electrochemistry. The Ru (2+) MV (2+)-Np guest and CB[8] host can form a stable 1:1 inclusion complex, in which the naphthalene residue is back-folded and inserted together with the viologen residue into the cavity of CB[8]. The selective binding of Ru (2+)-MV (2+)-Np guest with beta-CD and CB[8] host is also investigated. We find that CB[8] binds the Ru (2+)-MV (2+)-Np guest stronger than beta-CD. Upon light irradiation, a MV (+*) radical cation stabilized in the cavity of CB[8] accompanied by the naphthalene residue has been observed. This novel system may open a new way for design and synthesis of photoactive molecular devices. PMID- 18419160 TI - Highly diastereoselective synthetic route to enantiopure beta(2)-amino acids and gamma-amino alcohols using a fluorinated oxazolidine (Fox) as chiral auxiliary. AB - The alkylation reactions of an amide enolate derived from a trifluoromethylated oxazolidine (Fox) chiral auxiliary occur with a complete diastereoselectivity and in good yields with various electrophiles. This reaction provides a versatile and straightforward strategy for the synthesis of beta(2)-amino acids and gamma-amino alcohols in enantiopure form. PMID- 18419161 TI - Taking advantage of the radical character of tris(2,4,6-trichlorophenyl)methyl to synthesize new paramagnetic glassy molecular materials. AB - This paper describes the synthesis of the novel bis[4-(N-carbazolyl)-2,6 dichlorophenyl](2,4,6-trichlorophenyl)methyl radical (2*) and tris[4-(N carbazolyl)-2,6-dichlorophenyl]methyl radical (3*). A Friedel-Crafts reaction on [4-(N-carbazolyl)-2,6-dichlorophenyl)bis(2,4,6-trichlorophenyl]methyl radical (1*), 2*, and 3* leads to the introduction of acyl chains in the 3- and 6 positions of the carbazolyl moiety without impairment of the radical character of the molecule to give radicals 5*, 6*, and 7*. All of these novel radical adducts are thermally stable, 5* and 6* being amorphous solids by differential scanning calorimetry. Electron paramagnetic resonance spectra of them show a multiplet at low temperature due to the electron-coupling with six aromatic hydrogens. They show electrochemical amphotericity being reduced and oxidized to their corresponding stable anionic and cationic species, respectively. These radical adducts have luminescent properties covering the red spectral band of the emission with high intensities. PMID- 18419162 TI - Microwave-assisted efficient and convenient synthesis of 2,4(1H,3H) quinazolinediones and 2-thioxoquinazolines. AB - An efficient and convenient method was developed for the preparation of 2,4(1H,3H)-quinazolinediones and 2-thioxoquinazolinones. Substituted methyl anthranilate reacted with various iso(thio)cyanates in DMSO/H2O without any catalyst or base by using microwave irradiation to generate diversity on the 2,4(1H,3H)-quinazolinediones or 2-thioxoquinazolinones. A variety of substrates can participate in the process with good yields and high purities, making this methodology suitable for library synthesis in drug discovery efforts. PMID- 18419163 TI - Reagent-based, modular, tandem Michael approach for obtaining different indoline alkaloid-inspired polycyclic architectures. AB - A modular, reagent-based approach to obtain different indoline alkaloid-inspired, tetracyclic architectures is developed. With the use of TBSOTf as a Lewis acid, we report here a tandem Michael-based approach that led to the synthesis of a diastereomeric mixture of tetracyclic derivatives with two additional six membered rings. By simply changing the Lewis acid to TMSOTf, we were able to obtain a different tetracyclic compound having additional functionalized 5- and 7 membered rings with complete stereocontrol. PMID- 18419164 TI - Regulation of SNARE-mediated membrane fusion during exocytosis. PMID- 18419166 TI - Excluded volume effects on the intrachain reaction kinetics. AB - On the basis of the recently developed optimized Rouse-Zimm theory of chain polymers with excluded volume interactions, we calculate the long-time first order rate constant k(1) for end-to-end cyclization of linear chain polymers. We first find that the optimized Rouse-Zimm theory provides the longest chain relaxation times tau(1) of excluded volume chains that are in excellent agreement with the available Brownian dynamics simulation results. In the free-draining limit, the cyclization rate is diffusion-controlled and k(1) is inversely proportional to tau(1), and the k(1) values calculated using the Wilemski-Fixman rate theory are in good agreement with Brownian dynamics simulation results. However, when hydrodynamic interactions are included, noticeable deviations are found. The main sources of errors are fluctuating hydrodynamic interaction and correlation hole effects as well as the non-Markovian reaction dynamic effect. The physical natures of these factors are discussed, and estimates for the magnitudes of required corrections are given. When the corrections are included, the present theory allows the prediction of accurate k(1) values for the cyclization of finite-length chains in good solvents as well as the correct scaling exponent in the long-chain limit. PMID- 18419167 TI - Selection rules and symmetry relations for four-wave mixing measurements of uniaxial assemblies. AB - Uniaxial systems represent the next lowest symmetry below isotropic and are ubiquitous. The objective of the present work is to present a systematic foundation for interpreting polarization-dependent four-wave mixing measurements of oriented and aligned assemblies. Orientational averages connecting the molecular frame to the macroscopic frame in uniaxial assemblies were derived for several common molecular symmetry groups for coherent anti-Stokes Raman spectroscopy (CARS) measurements, coherent anti-Stokes two-photon spectroscopy (CATS) probing electronic transitions, resonant two-photon absorption (2PA), and traditional Raman measurements. First, the complete set of orientational averages connecting the molecular and macroscopic frames was compiled for the most general case of C1 molecular symmetry. Then, the orientational averages of a select few commonly occurring molecular symmetry groups (Cs, C2, C2v, and C3v) were explored in greater detail to illustrate the approach and to facilitate the interpretation of routine experimental measurements. One outcome of this analysis is the prediction of efficient electric dipole-allowed chiral-specific four-wave mixing in uniaxially oriented media. PMID- 18419170 TI - Acute lethal and teratogenic effects of tributyltin chloride and copper chloride on mahi mahi (Coryphaena hippurus) eggs and larvae. AB - Acute and chronic bioassays were used to evaluate the lethal and sublethal effects of copper chloride and tributyltin chloride on mahi mahi (Coryphaena hippurus) embryos, a pelagic life stage often found in the surface microlayer where anthropogenic contaminants can accumulate. Acute bioassay testing determined the median lethal concentration (LC50) for the test organism after 48 h of exposure. Chronic toxicity tests were used to determine the measurement of sublethal parameters, such as developmental abnormalities after 72-h exposures to the toxicants. Embryos were collected 4 h postfertilization and subsequently exposed to 1, 5, 25, 50, and 100 microg/L of copper chloride and 3, 10, 20, 30, and 50 microg/L of tributyltin chloride. Analysis of hatch rate percentage determined that the mean 48-h LC50 of copper chloride and tributyltin chloride was 32.8 and 16.7 microg/L, respectively, based on the pooled data of four experiments with four replicates for each metal. Consistent abnormalities, such as yolk sac swelling, spinal deformities, and decreased hatch rates, were observed for each metal. Teratogenic responses to copper chloride and tributyltin chloride demonstrate the need to investigate further the impacts of pollution in the open oceans to a species indigenous to, and commercially important to, the Florida (USA) coasts. This information could then lead to the future development of a surface microlayer bioassay using mahi mahi embryos. PMID- 18419168 TI - Characterization of the nanostructure of complexes formed by a redox-active cationic lipid and DNA. AB - We report characterization of the nanostructures of complexes formed between the redox-active lipid bis(n-ferrocenylundecyl)dimethylammonium bromide (BFDMA) and DNA using small-angle neutron scattering (SANS) and cryogenic transmission electron microscopy (cryo-TEM). A particular focus was directed to the influence of lipid oxidation state (where reduced BFDMA has a net charge of +1 and oxidized BFDMA has a charge of +3) on the nanostructures of the solution aggregates formed. Complexes were characterized over a range of charge ratios of reduced BFDMA to DNA (1.1:1, 2.75:1, and 4:1) in solutions of 1 mM Li2SO4. For these complexes, a single peak in the SANS data at 1.2 nm(-1) indicated that a nanostructure with a periodicity of 5.2 nm was present, similar to that observed with complexes of the classical lipids DODAB/DOPE and DNA (multilamellar spacing of 7.0 nm). The absence of additional Bragg peaks in all the SANS data indicated that the periodicity did not extend over large distances. Both inverse Fourier transform analysis and form factor fitting suggested formation of a multilamellar vesicle. These results were confirmed by cryo-TEM images in which multilamellar complexes with diameters between 50 and 150 nm were observed with no more than seven lamellae per aggregate. In contrast to complexes of reduced BFDMA and DNA, Bragg peaks were absent in SANS spectra of complexes formed by oxidized BFDMA and DNA at all charge ratios investigated. The low-q behavior of the SANS data obtained using oxidized BFDMA and DNA complexes suggested that large, loose aggregates were formed, consistent with complementary cryo-TEM images showing predominantly loose disordered aggregates. Some highly ordered spongelike and cubic phase nanostructures were also detected in cryo-TEM images. We conclude that control of BFDMA oxidation state can be used to manipulate the nanostructures of lipid-DNA complexes formed using BFDMA. PMID- 18419171 TI - Creating metal-spiked bed sediments: a case study from Orewa estuary, New Zealand. AB - Spiking sediments to achieve target concentrations of heavy metal pollutants is a key step in sediment toxicity tests. It is difficult, however, to ensure that metals in an artificially spiked sediment will behave naturally. A method has been developed in the present study to create Cu-, Pb-, and Zn-spiked sediments in which naturally occurring adsorption onto sediment surfaces is the dominant process binding the metals and in which precipitation of readily redissolved minerals and other metal-bearing phases (artifacts of the spiking procedure) are avoided. Uncontaminated bed sediment from an intertidal mudflat in the Orewa estuary, New Zealand, was characterized in terms of existing metal content, optimal adsorption pH, and adsorption capacity. Competitive adsorption between Cu and Pb as well as complexation by seawater anions only slightly affected metal adsorption from seawater. Surface complexation modeling indicated that iron oxide surfaces in the sediment likely were dominating metal adsorption processes. Spiking experiments were designed using these established adsorption characteristics but with significantly higher (>100-fold) concentrations of sediments and dissolved metals and a liquid to solid (L:S) ratio of approximately 5.5. An equilibration time of at least 36 h was required to achieve a reproducible target metal concentration, which could be reliably predicted from the L:S ratio and the initial metal concentration in the spiking solution. Adsorption equilibrium remained the process governing metal binding to the sediment, and no indication was observed that the adsorption capacity of the sediment had been exceeded or that additional metal-bearing phases had been formed. PMID- 18419172 TI - Kinetic study of the 2-methyl-3-methoxy-4-phenylbutanoic acid produced by oxidation of microcystin in aqueous solutions. AB - Microcystins (MCs) are a family of related cyclic hepatotoxic heptapeptides, of which more than 70 types have been identified. The chemically unique nature of the C20 beta-amino acid, (2S,3S,8S,9S)-3-amino-9-methoxy-2,6,8-trimethyl-10 phenyldeca-4,6-dienoic acid (Adda), portion of the MCs has been exploited to develop a strategy to analyze the entirety. Oxidation of MCs causes the cleavage of MC Adda to form 2-methyl-3-methoxy-4-phenylbutanoic acid (MMPB). In the present study, we investigated the kinetics of MMPB produced by oxidation of the most-often-studied MC variant, MC-LR (L = leucine, R = arginine), with permanganate-periodate. This investigation allowed insight regarding the influence of the reaction conditions (concentration of the reactants, temperature, and pH) on the conversion rate. The results indicated that the reaction was second order overall and first order with respect to both permanganate and MC-LR. The second-order rate constant ranged from 0.66 to 1.35 M/s at temperatures from 10 to 30 degrees C, and the activation energy was 24.44 kJ/mol. The rates of MMPB production can be accelerated through increasing reaction temperature and oxidant concentration, and sufficient periodate is necessary for the formation of MMPB. The initial reaction rate under alkaline and neutral conditions is higher than that under acidic conditions, but the former decreases faster than the latter except under weakly acidic conditions. These results provided new insight concerning selection of the permanganate-periodate concentration, pH, and temperature needed for the oxidation of MCs with a high and stable yield of MMPB. PMID- 18419173 TI - Fate of the nonsteroidal anti-inflammatory drug naproxen in agricultural soil receiving liquid municipal biosolids. AB - Naproxen (2-(6-methoxy-2-naphthyl) propionic acid) is widely used for the treatment of pain and swelling associated with arthritis, gout, and other inflammatory conditions. Naproxen has been detected in municipal sewage outflows and in surface waters and could reach agricultural land through the application of municipal biosolids or reclaimed water. The persistence characteristics of naproxen in three agricultural soils were investigated. In laboratory microcosms of moist soil incubated at 30 degrees C, [O-14CH3]naproxen was rapidly and thoroughly mineralized to 14CO2 with comparable kinetics in a sandy loam soil, a loam soil, and a silt loam soil. Naproxen mineralization was responsive to soil temperature and soil moisture content, consistent with the primary mechanism of dissipation being biodegradation. Mineralization of naproxen was hastened by the addition of liquid municipal biosolids (LMBs) from a municipal sewage treatment plant that aerated this material. Naproxen was stable in autoclaved soils with or without addition of autoclaved LMBs, whereas naproxen was rapidly mineralized in sterile soil supplemented with nonsterile LMBs. An enrichment culture was obtained from aerobically digested LMBs in a mineral salts medium with naproxen as the sole source of carbon. The culture converted the parent compound to the corresponding naphthol, O-desmethyl naproxen. In summary, naproxen was rapidly removed from soil, with mesophilic aerobic biodegradation being the primary mechanism of dissipation. Microorganisms carried in biosolids enhanced naproxen dissipation in soil, with the initial mechanism of attack likely being O demethylation. We conclude on this basis that naproxen in soils receiving biosolids would be readily biodegradable and, in the absence of preferential flow or runoff, pose little risk for contamination of adjacent water or crops. PMID- 18419174 TI - Chemical availability and sediment toxicity of pyrethroid insecticides to Hyalella azteca: application to field sediment with unexpectedly low toxicity. AB - Tenax extraction is a simple, inexpensive approach to estimate the bioavailability of hydrophobic organic contaminants from sediment. In the present study, a single-point Tenax extraction was evaluated regarding its correlation with the acute toxicity to Hyalella azteca using field-collected sediments in California, USA. Pyrethroids were believed to be the primary contributor to the observed toxicity, and a significant correlation existed between the expected toxicity (given pyrethroid concentrations) and the mortality at most sampling sites. A small subset of sites, however, showed unexpectedly low toxicity to H. azteca despite high concentrations of pyrethroids. These samples were evaluated by Tenax extraction with the expectation that this procedure, which qualifies bioavailable instead of total pyrethroid concentration in sediment, would better explain the anomalously low toxicity. The term bioavailable toxic unit was proposed to link sediment toxicity with chemical availability, and the toxicity in the 17 selected sediments was better explained using Tenax extraction. The r2 value of the regression between sediment toxicity and toxic unit for the 17 sediments increased from 0.24 to 0.60 when the Tenax-extractable concentration was used in place of the total concentration. Results also showed that adsorption to sand particles might play a controlling role in pyrethroid bioavailability and, in turn, sediment toxicity to benthic invertebrates. PMID- 18419175 TI - Concentrations and patterns of perfluoroalkyl acids in Georgia, USA surface waters near and distant to a major use source. AB - Perfluoroalkyl acids (PFAAs) are widespread contaminants emanating from, among other sources, the production/degradation of fluorinated chemicals used in surface repellant applications, such as carpet manufacturing. The goal of the present study was to assess the concentrations of PFAAs, including perfluorooctane sulfonate (PFOS), perfluorooctanoic acid (PFOA), perfluorononanoic acid (PFNA), perfluorodecanoic acid (PFDA), perfluoroundecanoic acid (PFUA), and perfluorooctane sulfonamide (PFOSA), in surface waters both near a wastewater land application system (LAS) in Dalton (GA, USA), home to North America's largest carpet manufacturing site, and distant to this location (Altamaha River, GA, USA) to understand the fate of PFAAs in freshwater. Levels of PFAAs were high in the Conasauga River (GA, USA) downstream of the LAS (PFOA, 253-1,150 ng/L; PFOS, 192-318 ng/L; PFNA, 202-369 ng/L; PFDA, 30.1-113 ng/L; PFUA, 58.0-99.2 ng/L; PFOSA, 162-283 ng/L) and in streams and ponds in Dalton (PFOA, 49.9-299 ng/L; PFOS, 15.8-120 ng/L), and were among the highest measured at a nonspill or direct-release location. Perfluoroalkyl acids in the Altamaha River were much lower (PFOA, 3.0-3.1 ng/L; PFOS, 2.6-2.7 ng/L), but were a source of PFAAs to Georgia's estuaries. A preliminary hazard assessment indicated that concentrations of PFOS at two sites in the Conasauga River exceeded the threshold effect predicted for birds consuming aquatic organisms that are exposed continuously to the PFOS levels at these sites. Assuming that toxicity for all PFAAs quantified is equal to that of PFOS, the sum total PFAAs at two sites within the Conasauga River exceeded PFOS thresholds for aquatic and avian species, warranting additional research. PMID- 18419176 TI - Stress response of mammalian cells incubated with landfill leachate. AB - Environmental contamination from landfill leachate has been linked to disturbances in human health. Often, however, only global parameters, such as dissolved organic content, chemical oxygen demand, and 5-d biological oxygen demand, are used to evaluate wastewater quality. In the present study, we determined leachate cytotoxicity and stress response of leachate-treated mammalian cells using in vitro bioassays and other molecular techniques. The modified E-screen assay using human breast cancer MCF-7 cells was used to determine the estrogenic potential and/or cytotoxicity of water samples from two solid-waste landfills in Tunisia. The cytotoxicity mechanism of the leachate was determined by DNA fragmentation and lactate dehydrogenase assays. The stress response of heat shock protein (HSP) 47-positive Chinese hamster ovary cells treated with leachate also was determined. Proteomics analyses and real-time reverse transcription-polymerase chain reaction were used to determine and confirm the enhanced expressions of certain stress-related proteins. Results showed that the leachate samples generally did not have estrogenic activity. Instead, they were cytotoxic toward MCF-7 cells, and the cytotoxicity was by necrosis during the early stages of incubation. Leachate also enhanced the expression of HSP and various stress-related proteins, such as heterogeneous nuclear ribonucleoprotein E1, phosphoglycerate mutase, and nuclear matrix protein 200, in MCF-7 cells. These can be considered as survival mechanisms against leachate-induced cytotoxicity. PMID- 18419177 TI - Spatially distributed ecological risk for fish of a coastal food web exposed to dioxins. AB - The ecological risk posed by 2,3,7,8-polychlorodibenzo-p-dioxins and furans (PCDD/Fs) and dioxin-like polychlorobiphenyl (PCB) congeners to five edible fish species of the aquatic food web of Venice Lagoon, Italy, was estimated by applying a state-of the-art kinetic bioaccumulation model. Site-specific data were used to define a representative food web. The experimental data set for model validation and application included PCB and PCDD/F congener concentrations in sediments, in water, and in five organisms (both invertebrates and fish). The spatial distribution of risk was evaluated by dividing the lagoon into six homogeneous areas, and for each area, sediment, water, and organism dioxins concentrations were calculated. The bioaccumulation model was calibrated for both nonmetabolizing and metabolizing congeners, the metabolic elimination rates of which were estimated. The model validation showed an acceptable bioaccumulation estimation, evaluated using the model bias parameter. The calibrated model was applied to the six areas of the lagoon to estimate the fish predicted exposure concentration as 2,3,7,8-tetrachlorodibenzo-p-dioxin toxicity equivalents from sediment concentration. Internal no-effect concentrations were calculated for each fish species from literature data. Risk was estimated by applying the hazard quotient (HQ) approach, obtaining the ecological risk for each fish species on the basis of 90 and 99% protection levels, in each of the six lagoon areas. The sediment dioxins concentration does not pose a significant risk to the selected fish species at the 90% protection target (HQ<1), whereas risk is significant (HQ>1) at the 99% protection target. Risk results were higher near the Porto Marghera industrial district, Italy, and in lagoon zones characterized by a low water-exchange rate and freshwater basin inputs. PMID- 18419178 TI - Influence of pH on the acute toxicity of ammonia to juvenile freshwater mussels (fatmucket, Lampsilis siliquoidea). AB - The objective of the present study was to evaluate the influence of pH on the toxicity of ammonia to juvenile freshwater mussels. Acute 96-h ammonia toxicity tests were conducted with 10-d-old juvenile mussels (fatmucket, Lampsilis siliquoidea) at five pH levels ranging from 6.5 to 9.0 in flow-through diluter systems at 20 degrees C. Acute 48-h tests with amphipods (Hyalella azteca) and 96 h tests with oligochaetes (Lumbriculus variegatus) were conducted concurrently under the same test conditions to determine the sensitivity of mussels relative to these two commonly tested benthic invertebrate species. During the exposure, pH levels were maintained within 0.1 of a pH unit and ammonia concentrations were relatively constant through time (coefficient of variation for ammonia concentrations ranged from 2 to 30% with a median value of 7.9%). The median effective concentrations (EC50s) of total ammonia nitrogen (N) for mussels were at least two to six times lower than the EC50s for amphipods and oligochaetes, and the EC50s for mussels decreased with increasing pH and ranged from 88 mg N/L at pH 6.6 to 0.96 mg N/L at pH 9.0. The EC50s for mussels were at or below the final acute values used to derive the U.S. Environmental Protection Agency's acute water quality criterion (WQC). However, the quantitative relationship between pH and ammonia toxicity to juvenile mussels was similar to the average relationship for other taxa reported in the WQC. These results indicate that including mussel toxicity data in a revision to the WQC would lower the acute criterion but not change the WQC mathematical representation of the relative effect of pH on ammonia toxicity. PMID- 18419179 TI - Response of a New Zealand mayfly (Deleatidium spp.) to acid mine drainage: implications for mine remediation. AB - Investigating the toxicity of acid mine drainage (AMD) on benthic communities in receiving waters can be highly challenging because of the difficulty in unraveling the effects of acidity, dissolved metals, and precipitates. Furthermore, the survival of different species may vary depending on any natural adaptation they may have acquired to low pH, metals, or sedimentation. We investigated the effect of different pHs and AMD on the survival of a common New Zealand leptophlebiid mayfly (Deleatidium spp.) in 96-h laboratory trials. Our results indicate that the primary driver of toxicity in AMD was pH, although some mortality could be attributable to the presence of dissolved heavy metals at low pH (90%) for lizards in all treatments. Changes in food consumption and body weight were observed at 35 and 45 mg/kg/d. Alterations in hematological end points; blood chemistries (albumin, total protein, alkaline phosphatase, calcium); kidney, spleen, and liver weights; and adverse histopathology were observed in lizards exposed at 25 to 45 mg/kg/d. Testosterone concentration, sperm count, and motility were variable between treatments. Although not significant, incidences of hypospermia and testicular atrophy were observed in some individuals. Together, these data suggest a lowest-observed-adverse effect level of 25 mg/kg/d and a no-observed adverse effect level of 15 mg/kg/d in S. occidentalis. PMID- 18419184 TI - Evaluating the trophic transfer of cadmium, polonium, and methylmercury in an estuarine food chain. AB - We examined the transfer of Cd, methylmercury (MeHg), and Po in an estuarine food chain (from phytoplankton to zooplankton [Daphnia pulex] to killifish [Fundulus heteroclitus] and, finally, to juvenile striped bass [Morone saxatilis]) to better understand both the extent to which these elements may biomagnify and the underlying mechanisms governing this biomagnification. Among the phytoplankton examined (Cyclotella meneghiniana and Chlamdomonas reinhardtii), metal uptake was rapid, and volume concentration factors for all metals reached values between 10(4) and 10(5). The resulting assimilation efficiencies (AEs) of ingested metals in all animals were highest for MeHg, with values greater than 76%. The AEs of Cd were 21 to 33% in D. pulex, 1 to 16% in F. heteroclitus, and 38 to 56% in M. saxatilis. Polonium AEs were 69 to 87% in D. pulex, 25 to 41% in F. heteroclitus, and 9 to 21% in M. saxatilis. Loss rate constants (ke) of metals in D. pulex ranged from 0.04/d for MeHg to 0.39/d for Po; metal ke values for F. heteroclitus ranged from 0.01 to 0.02/d. Using a kinetic model, we showed that the trophic transfer factor, defined as the ratio of metal concentration in predatory animals to metal concentration in prey organisms, was greater than unity for all three metals in D. pulex feeding on phytoplankton, suggesting that these metals may be biomagnified at this trophic step. In killifish feeding on D. pulex, the trophic transfer factor was consistently greater than one for MeHg, consistently less than one for Cd, and from 0.1 to 1.4 for Po, suggesting that both MeHg and Po have the potential to biomagnify at this trophic step. PMID- 18419185 TI - "What exactly are you inferring?" A closer look at hypothesis testing. AB - This critical review describes the confused application of significance tests in environmental toxicology and chemistry that often produces incorrect inferences and indefensible regulatory decisions. Following a brief review of statistical testing theory, nine recommendations are put forward. The first is that confidence intervals be used instead of hypothesis tests whenever possible. The remaining recommendations are relevant if hypothesis tests are used. They are as follows: Define and justify Type I and II error rates a priori; set and justify an effect size a priori; do not confuse p(E|H0) and p(H0|E); design tests permitting Positive Predictive Value estimation; publish negative results; estimate a priori, not post hoc, power; as warranted by study goals, favor null hypotheses that are not conventional nil hypotheses; and avoid definitive inferences from isolated tests. PMID- 18419186 TI - Avoidance tests in site-specific risk assessment--influence of soil properties on the avoidance response of Collembola and earthworms. AB - The ability of organisms to avoid contaminated soils can act as an indicator of toxic potential in a particular soil. Based on the escape response of earthworms and Collembola, avoidance tests with these soil organisms have great potential as early screening tools in site-specific assessment. These tests are becoming more common in soil ecotoxicology, because they are ecologically relevant and have a shorter duration time compared with standardized soil toxicity tests. The avoidance response of soil invertebrates, however, can be influenced by the soil properties (e.g., organic matter content and texture) that affect behavior of the test species in the exposure matrix. Such an influence could mask a possible effect of the contaminant. Therefore, the effects of soil properties on performance of test species in the exposure media should be considered during risk assessment of contaminated soils. Avoidance tests with earthworms (Eisenia andrei) and springtails (Folsomia candida) were performed to identify the influence of both organic matter content and texture on the avoidance response of representative soil organisms. Distinct artificial soils were prepared by modifying quantities of the standard artificial soil components described by the Organization for Economic Co-operation and Development to achieve different organic matter and texture classes. Several combinations of each factor were tested. Results showed that both properties influenced the avoidance response of organisms, which avoided soils with low organic matter content and fine texture. Springtails were less sensitive to changes in these soil constituents compared with earthworms, indicating springtails can be used for site-specific assessments of contaminated soils with a wider range of respective soil properties. PMID- 18419187 TI - Ozone exposure affects feeding and locomotor behavior of adult Bufo marinus. AB - Ozone (O3), a reactive component of air pollution, depresses feeding and voluntary locomotor behavior in laboratory rodents, but the effects of O3 on amphibian behavior are not known. We evaluated the effects of 4 h of exposure to air or ozone (0.6 microl/L), on two ecologically relevant behaviors of the toad Bufo marinus. Toads were offered five mealworms at 1, 24, and 48 h after exposure. One hour after exposure, O3-exposed toads ate fewer mealworms than did air-exposed toads (Fisher exact test, p=0.005). Within 24 h after exposure, all toads ate four or five mealworms. Because movement is a key component of toad feeding behavior, we tested additional toads (n=25) for voluntary locomotor behavior during three 1-h trials in a 2.9-m2 open-field arena. Mean (+/-standard deviation) total distance moved was: pre-exposure, 29+/-19.5 m; 1-h postexposure, 13+/-15.6 m; and 24-h postexposure, 17+/-17.4 m. The means were not statistically different by repeated measures analysis of covariance. Therefore, our results suggest that a single 4-h exposure to O3 depresses toad feeding behavior after exposure but had little effect on voluntary locomotor behavior. PMID- 18419188 TI - Enhanced reactivity of hydroxylated polycyclic aromatic hydrocarbons to birnessite in soil: reaction kinetics and nonextractable residue formation. AB - Phenanthrene and pyrene were not transformed by birnessite (delta-MnO2) in the presence of phenol. The phenoxy radicals generated from phenol by birnessite did not act as a mediator for polycyclic aromatic hydrocarbon radical reaction under the studied conditions. In contrast, 9-hydroxyphenanthrene and 1-hydroxypyrene were remarkably sensitive to birnessite. The disappearance patterns of the test compounds both in the aqueous phase and soil followed first-order kinetics, with a linear relationship found between the rate constants and the surface area of birnessite. Moreover, the data indicated that the reaction was faster in the presence of soil than in the aqueous phase probably because of the presence of hydroxyl groups in soil organic matter. Sequential solvent extraction was not successful in the recovery of 9-hydroxyphenanthrene from the birnessite-treated soil samples, and capillary electrophoresis data suggest the formation of nonextractable residues of the compound in soil. In addition, the acute toxicity determined by Microtox declined approximately 8.3 times in the soil samples treated with birnessite compared to untreated samples, demonstrating that the toxic compound was no longer present as its parent form. PMID- 18419189 TI - Effects of estimates from different geochemical models on metal fate predicted by coupled speciation-fate models. AB - Coupled metal speciation-fate models are an improvement over stand-alone fate transport models for accurately assessing metal fate and transport. These coupled models estimate fate-controlling partition coefficients using geochemical speciation/complexation models. Commercially available geochemical models are practical options for a two-step, loose coupling with fate-transport models. These models differ in their partitioning estimates because of differences in assumptions, databases, and so on. The present study examines the effects of differences in estimates from geochemical models on estimates of cationic metal fate using two geochemical models: the Windermere humic aqueous model (WHAM) and the minicomputer equilibrium+ model (MINEQL+). The results from each geochemical model were used as input to the fate module of TRANSPEC (a general, coupled metal transport and speciation model). The two versions of the TRANSPEC model were then used to assess the fate of five cationic metals (Cd, Cu, Ni, Pb, and Zn) in Ross Lake (Flin Flon, MB, Canada; alkaline, eutrophic, mine impacted), Kelly Lake (Sudbury, ON, Canada; circumneutral, mesotrophic, mine influenced), and Lake Tantare (Quebec City, QC, Canada; acidic, oligotrophic, pristine). For relatively soluble metals (Cd, Ni, and Zn), the WHAM and MINEQL+ estimates of speciation/complexation were similar for Ross and Kelly lakes but differed for Lake Tantare. These differences, however, did not result in significant differences in overall fate estimates. Marked differences were observed between the WHAM and MINEQL+ estimates of partition coefficient, Kd, for more particle reactive Cu and Pb that translated into the greatest impact on fate in mesotrophic Kelly Lake, in which particle movement is important for fate. PMID- 18419190 TI - High molecular weight petroleum hydrocarbons differentially affect freshwater benthic macroinvertebrate assemblages. AB - High-molecular weight petroleum hydrocarbons (HMWPHs) are a common pollutant in urban freshwater sediments. A previous study found that HMWPHs derived from synthetic oils are detrimental to aquatic benthic macroinvertebrates at concentrations greater than 840 mg/kg, but it is unclear what effect hydrocarbons derived from other sources have on these organisms. A field-based microcosm experiment was conducted to determine whether natural and other types of HMWPHs produce similar effects on indigenous benthic macroinvertebrates as was induced by synthetic motor oils. Ordinations and comparisons of sensitive species indicated that HMWPHs from different anthropogenic sources negatively affected macroinvertebrates, whereas naturally occurring hydrocarbons above the threshold of 840 mg/kg had no detrimental effect. This result suggests that total petroleum hydrocarbon content, which is often used to identify hydrocarbon pollutants, is a good conservative indicator of HMWPH concentrations affecting the biota. PMID- 18419191 TI - Uses of subcellular metal distribution in prey to predict metal bioaccumulation and internal exposure in a predator. AB - In the present study, rock oysters (Saccostrea cucullata) were first exposed to cadmium and zinc for two weeks to modify their subcellular metal partitionings. The relationship between subcellular metal (Cd and Zn) partitioning in the oysters and metal bioaccumulation and fractionation in predatory gastropods (Thais clavigera) was then examined by feeding to the predator oysters that were preexposed to metal for two to four weeks. We also investigated the relationship between the PAM in the oysters and the biochemical biomarkers in the gastropods. Thais clavigera accumulated Cd effectively from their prey, but no correlation was found between the Cd body concentrations in T. clavigera and the internal metal partitioning in the prey. A significant positive correlation was found between the Cd in the trophically available metal (TAM) fraction of oysters and the Cd in the metal-sensitive fraction of T. clavigera and between the Cd in the TAM fraction of oysters and the metallothionein induction in whelks. Zinc was highly regulated by both S. cucullata and T. clavigera, and their Zn body concentrations remained constant throughout the exposure period. No relationship between Zn bioaccumulation and any of the subcellular fractions was found. The present study may lead to a better understanding of the dietary metal exposure mechanism. PMID- 18419192 TI - Effects of chronic dietary and waterborne cadmium exposures on the contamination level and reproduction of Daphnia magna. AB - Regulatory assessments of metal toxicity on freshwater organisms assume that toxic effects are caused by dissolved metals. In aquatic systems, organisms are exposed to both dissolved and particulate-bound metals. In this study, the chronic toxicity of dietary cadmium (Cd) on the reproduction and Cd body burden of Daphnia magna was investigated. Daphnids (<24 h) were successively exposed to dissolved Cd (8 h) and then to uncontaminated or contaminated algae (16 h) for 21 d. The results show a higher Cd burden in daphnids because of the addition of contaminated food and reveal that Cd uptake by D. magna from water and food was additive for the lowest Cd concentrations tested. Similar Cd distributions (cytosolic and insoluble fractions) were observed in the two groups of organisms, showing similar potential toxicity of Cd accumulated from the two exposure routes. Dietary Cd induces deleterious effects on D. magna reproduction. On the basis of Cd body burden of daphnids, the results support the claim that waterborne and dietary Cd exposures were additive in causing toxicity for Cd concentrations lower than 25 microg/L. At the highest Cd concentrations, the importance of dietary Cd on the daphnid contamination level decreases and confounding factors such as feeding rate reduction seem to appear, which induce an effect on neonate reproduction. In this study, we illustrate the need to take the dietary pathway into account in regulatory assessments and to establish effective concentrations with particulate-bound metals. PMID- 18419193 TI - Spatial covariation of microbial community composition and polycyclic aromatic hydrocarbon concentration in a creosote-polluted soil. AB - Little is known about the spatial connection between soil microbial community composition and polycyclic aromatic hydrocarbon (PAH) concentration. A spatially explicit survey at a creosote-contaminated site demonstrated that microbial biomass (total concentration of phospholipid fatty acids [PLFAs]) and microbial community composition (PLFA fingerprints) were spatially autocorrelated, mostly within a distance of 25 m, and covaried with PAH concentrations. The concentration of PLFAs indicative of gram-negative bacteria (16:1omega7c, 16:1omega7t, 18:1omega7, cy17:0, and cy19:0) increased in the PAH hot spots, whereas PLFAs representing fungi and gram-positive bacteria (including actinomycetes) were negatively correlated to PAH concentrations. Most PLFAs were spatially autocorrelated, with distances varying between 4 and 25 m. Those PLFAs that increased in PAH-contaminated soil had autocorrelation ranges between 4 and 16 m, whereas the fungal indicator PLFA 18:2omega6,9 had the largest autocorrelation range (25 m). Bacterial strains isolated using a spray-plate technique and with the same PLFA composition as that in contaminated soil samples were capable of degrading phenanthrene, fluoranthene, and pyrene, indicating that the main PAH degraders could be isolated. PMID- 18419194 TI - A fugacity approach for assessing the bioaccumulation of hydrophobic organic compounds from estuarine sediment. AB - The bioavailability of four sediment-spiked hydrophobic organic contaminants (HOCs; chrysene, benzo[a]pyrene, chlordane, and Aroclor 1254) was investigated by comparing bioaccumulation by the amphipod Corophium colo with uptake into a thin film of ethylene/vinyl acetate (EVA) copolymer. The EVA thin film is a solid phase extraction medium previously identified as effective at measuring the bioavailable contaminant fraction in sediment. The present study presents the results of 11 separate treatments in which chemical uptake into EVA closely matched uptake into lipid over 10 d. For all compounds, the concentration in EVA was a good approximation for the concentration in lipid, suggesting that this medium would be an appropriate biomimetic medium for assessing the bioaccumulation of HOCs during risk assessment of contaminated sediment. For chrysene and benzo[a]pyrene, limitations on bioaccumulation and toxicity because of low aqueous solubility were observed. The fugacity of the compounds in lipid (flip) and in the EVA thin film (fEVA) also was determined. The ratio of flip to fEVA was greater than one for all chemicals, indicating that all chemicals biomagnified over the duration of the exposure and demonstrating the potential for EVA thin-film extraction to assess trophic transfer of HOCs. PMID- 18419195 TI - Growth-inhibiting effects of 12 antibacterial agents and their mixtures on the freshwater microalga Pseudokirchneriella subcapitata. AB - The growth-inhibiting and binary joint effects of 12 antibacterial agents on the freshwater green alga Pseudokirchneriella subcapitata (Korschikov) Hindak were investigated over 72-h exposures. The toxicity values (the median inhibitory concentration value, in micromoles) in decreasing order of sensitivity were triclosan (0.0018)>triclocarban (0.054)>roxithromycin (0.056)>clarithromycin (0.062)>tylosin (0.20)>tetracycline (2.25)>chlortetracycline (3.49)>norfloxacin (5.64)>sulfamethoxazole (7.50)>ciprofloxacin (20.22)>sulfamethazine (31.26)>trimethoprim (137.78). Several of these antibacterial compounds would be toxic at the micrograms per liter concentrations reported in surface waters and sewage effluents. Simple additive effects were observed in binary mixtures of sulfonamides, and most tylosin, triclosan, or triclocarban combinations. Potentially synergistic effects were observed in binary mixtures of the same class, such as macrolides, tetracyclines, and fluoroquinolones, as well as in some combined drugs, such as trimethoprim and sulfonamides or tylosin and tetracyclines. Potentially antagonistic effects were only observed between tylosin and triclocarban, triclosan and norfloxacin, and triclocarban and norfloxacin. Although present at low concentrations in the aquatic environment, mixtures of these antibacterial agents can potentially affect algal growth in freshwater systems due to their combined action. PMID- 18419196 TI - Biodegradability of highly ethoxylated nonionic surfactants: determination of intermediates and pathways of biodegradation. AB - Existing data regarding alcohol ethoxylate (AE) surfactants indicate that structures with greater than 20 ethoxylate (EO) units per molecule or with multibranched alkyl chains may not pass a ready biodegradability test. This could have important consequences for complying with regional regulatory requirements and for the potential risks these chemicals could present to the environment. We investigated the influence of chemical structure on the biodegradability of AEs with different alkyl chain branching and EO content. The AEs investigated were a multibranched AE (average, 18 EO), an oxo-AE (monobranched; average, 23 EO), and a linear AE of oleochemical origin (average, 40 EO). The aims of the present study were to assess the ready biodegradability of AEs with high EO content and to establish the mechanism or pathway by which biodegradation occurs for the oxo AE. Biodegradation studies were conducted using standard test conditions (International Standards Organization 14593). Solid-phase extraction and liquid chromatography with electrospray mass spectrometry were used to detect profiles in both derivatized and underivatized extracts of samples. Derivatization with phthalic anhydride was used to improve ionization of the lower-ethoxylated AEs and free alcohol that were key indicators in the present study. All AEs were rapidly biodegraded, achieving more than 60% mineralization of parent material. Central fission was the predominant mechanism for the oxo-AE, as confirmed by identification of the oligomeric distribution and quantification of polyethylene glycol released during biodegradation. PMID- 18419197 TI - Estrogenic activity of lambda-cyhalothrin in the MCF-7 human breast carcinoma cell line. AB - Synthetic pyrethroids are widely used in both agricultural and urban environments for insect control. Lambda-cyhalothrin (LCT) is one of the most common pyrethroids and is used mainly for controlling mosquitoes, fleas, cockroaches, flies, and ants around households. Previous studies have addressed the environmental behaviors and acute toxicities of LCT, but little is known about its chronic toxicity, such as estrogen-like activity. In the present study, the estrogenic potential of LCT was evaluated using the MCF-7 human breast carcinoma cell line. The in vitro E-screen assay showed that 10(-7) M LCT could significantly promote MCF-7 cell proliferation, with a relative proliferative effect ratio of 45%. The cell proliferation induced by LCT could be blocked completely, however, by the addition of 10(-9) M of the estrogen receptor (ER) antagonist ICI 182,780. The semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) results showed that the Trefoil factor 1 (pS2) and progesterone receptor gene expression were up-regulated by 10(-7) M LCT for 2- and 1.5-fold, respectively. On the other hand, RT-PCR, Western blot analysis, and immunofluorescent assay demonstrated that LCT significantly repressed the mRNA and protein expression levels of ERalpha and ERbeta. These observations indicate that LCT possesses estrogenic properties and may function as a xenoestrogen, likely via a mechanism similar to that of 17beta-estradiol. The endocrine disruption potential of LCT should be considered when assessing the safety of this compound in sensitive environmental compartments. PMID- 18419198 TI - Dietary accumulation, disposition, and metabolism of technical pentabrominated diphenyl ether (de-71) in pregnant mink (Mustela vison) and their offspring. AB - Concentrations of polybrominated diphenyl ethers (PBDEs) in humans and wildlife suggest significant bioaccumulation potential in mammals, but no quantitative biomagnification data from controlled experiments are currently available. As part of a larger study examining the effects of PBDEs on mink (Mustela vison) reproduction and development, we examined congener-specific tissue distribution, maternal transfer, biotransformation, and biomagnification of the technical penta BDE mixture, DE-71, in farmed mink. Adult female mink were fed one of four concentrations of DE-71 (0-2.5 microg/g) in the diet from breeding through gestation and until weaning at 6 weeks postparturition. Parent PBDEs were measured in tissues and excreta of adult mink, kits, and juveniles, whereas hydroxylated PBDEs (OH-PBDEs) were measured in juveniles only. Similar lipid normalized concentrations of PBDEs were detected in most tissues of adult mink with the exception of brain, in which concentrations were significantly lower. Kits, however, had a higher proportion of PBDEs in brain compared with adults, presumably because of incomplete development of the blood-brain barrier. Maternal transfer favored lower-brominated PBDE congeners, and the bulk of the body burden in kits at weaning resulted from lactational rather than transplacental transfer. Lipid-normalized, whole-body biomagnification factors ranged from 0.5 to 5.2 for the major congeners and were highest for BDEs 47 and 153. Metabolism clearly limited the biomagnification of some PBDEs, and OH-PBDEs were detectable in plasma, liver, and feces. On a mass basis, OH-PBDEs accounted for 28 to 32% of the excreted fraction, indicating metabolism was an important elimination pathway. Further studies are required to understand the mechanisms of PBDE biotransformation. PMID- 18419199 TI - Protective role of vitamin E on the microcystin-induced oxidative stress in tilapia fish (Oreochromis niloticus). AB - Microcystins (MCs) are potent hepatotoxins produced by cyanobacteria in water systems that induce oxidative stress in fish. The present study investigated the effect of vitamin E pretreatment on MC-induced oxidative damage in the liver, kidneys, and gills of tilapia fish (Oreochromis niloticus). Groups of fish were fed vitamin E supplements (200 or 700 mg per kilogram of diet) for 7 d or received only commercial fish food and then were exposed to a single oral dose of cyanobacterial cells (120 microg of MC-LR [2:Leu, 4:Arg] per fish), and were sacrificed in 24 h. The potential benefits of vitamin E were evaluated based on lipid peroxidation (LPO), protein oxidation, catalase (CAT, Enzyme Commission [EC] 1.11.1.6), superoxide dismutase (EC 1.15.1.1), glutathione peroxidase (EC 1.11.1.9), glutathione reductase (EC 1.8.1.7), and the ratio of reduced glutathione to oxidized glutathione (GSH/GSSG). Microcystins induced a 1.6-fold increase in LPO values in liver, whereas vitamin E-pretreated fish showed no alteration on this biomarker. Enzyme activities levels were also ameliorated by the chemoprotectant, whereas protein oxidation and GSH/GSSG did not show any significant change. The higher vitamin E dose used proved to have the greater protective effects, particularly on the biomarkers LPO and CAT. The results show that vitamin E could have a potential use as a preventive or therapeutic measure in MC-exposed fish. PMID- 18419200 TI - Occurrence of endocrine-disrupting pesticide residues in wetland sediments from Beijing, China. AB - Wetlands are important habitats and spawning grounds to a diverse wildlife population and are particularly vulnerable to adverse effects of endocrine disruptors. Many pesticides possess hormonal activity and have thus been classified as endocrine disruptors. A new analytical method based on ultrasonic assisted extraction, solid-phase extraction, and gas chromatography technologies was developed for analysis of selected potential endocrine-disrupting pesticide residues in wetland sediment samples from northern Beijing, China. The newly developed method was a less time-consuming and less solvent-consuming way of analyzing residues without an air-drying process prior to extraction. The selected pesticides included the compounds hexachlorocyclohexane, cyclodiene, DDTs, chlordane, hexachlorobenzene, heptachlor, endrin aldehyde, heptachlor epoxide, dicofol, acetochlor, alachlor, metolachlor, chlorpyrifos, nitrofen, trifluralin, cypermethrin, fenvalerate, and deltamethrin. Characterization and identification of the selected endocrine-disrupting pesticide residues in sediments may help to assess current pollution status of endocrine-disrupting pesticides in the area. Total concentrations of pesticides ranged from 15.4 to 38.1 ng/g (dry weight) with a mean concentration of 23.7 ng/g (dry weight) for sediment samples. Although levels of all detected pesticides were below the New York State Department of Environmental Conservation, USA, sediment quality criteria, considering the potential health effects of endocrine disruptors even in a trace level, the use of the pesticides in the agricultural system should be minimized and regular monitoring is needed in the area. PMID- 18419201 TI - Distribution of toxins in various tissues of crucian carp intraperitoneally injected with hepatotoxic microcystins. AB - An acute toxicity experiment was conducted to examine the distribution and depuration of microcystins (MCs) in crucian carp (Carassius aurutus) tissues. Fish were injected intraperitoneally with extracted MCs at a dose of 200 microg MC-LR (where L=leucine and R=arginine) equivalent/kg body weight. Microcystin concentrations in various tissues and aquaria water were analyzed at 1, 3, 12, 24, and 48 h postinjection using liquid chromatography coupled with mass spectrometry. Microcystins were detected mainly in blood (3.99% of injected dose at 1 h), liver (1.60% at 1 h), gonad (1.49% at 3 h), and kidney (0.14% at 48 h). Other tissues, such as the heart, gill, gallbladder, intestine, spleen, brain, and muscle, contained less than 0.1% of the injected MCs. The highest concentration of MCs was found in blood (526-3,753 ng/g dry wt), followed by liver (103-1,656 ng/g dry wt) and kidney (279-1,592 ng/g dry wt). No MC-LR was detectable in intestine, spleen, kidney, brain, and muscle, whereas MC-RR was found in all examined fish tissues, which might result from organ specificity of different MCs. Clearance of MC-RR in brain tissue was slow. In kidney, the MC-RR content was negatively correlated with that in blood, suggesting that blood was important in the transportation of MC-RR to kidney for excretion. PMID- 18419202 TI - Environmental concentrations, fate, and risk assessment of pyrethrins and piperonyl butoxide after aerial ultralow-volume applications for adult mosquito management. AB - One of the most effective ways of managing adult mosquitoes that vector human and animals diseases is the use of ultralow-volume insecticides. Because of concerns about the safety of the insecticides used for the management of adult mosquitoes, we conducted an environmental fate and efficacy study in Princeton and Colusa (both CA, USA) after aerial applications of pyrethrins and piperonyl butoxide (PBO). One hour before application, PBO concentrations in water were 0.008 and 0.2175 microg/L for Princeton and Colusa, respectively. One hour after the spray event in Princeton, the average PBO concentrations were 0.0125 microg/cm2 on ground-deposition pads and 0.1723 microg/L in water samples, with concentrations decreasing significantly over time. One hour after the spray event in Colusa, the average PBO concentrations were 0.0199 microg/cm2 on deposition pads and 1.274 microg/L in water samples, with concentrations decreasing significantly over time. A significant time and location effect for both deposition pads and water samples in Princeton and Colusa was observed (p<0.001 and p=0.014, respectively). Pyrethrins were not detected in nearly all ground and water samples. One hour after application, mortality of Culex tarsalis and Culex pipiens in sentinel cages was significantly higher than at the control site for both locations (p<0.001). Risk quotients for aquatic surrogate species in Princeton and Colusa were 0.002 or less at 1 h after application, which did not exceed the U.S. Environmental Protection Agency risk quotient level of concern for endangered aquatic organisms of 0.05. Our results suggest that the amounts of pyrethrins and PBO deposited on the ground and in water after aerial ULV insecticide applications are lower than those estimated by previous exposure and risk assessments. PMID- 18419204 TI - Retrograde endoscopic-assisted percutaneous renal access: a novel "lasso" technique to achieve rapid secure access to the collecting system. AB - Image-guided percutaneous renal access for placement of an access sheath for percutaneous nephrolithotomy can be a challenging procedure, especially in patients with nondilated collecting systems, obstructed infundibula (stones or stricture), or extreme body habitus. We describe our experience using ureteroscopy along with a zero-tip stone basket to facilitate a through-and through (percutaneous-urethra) access to the collecting system. PMID- 18419205 TI - Digital nephroscopy: the next step. AB - BACKGROUND: The Hopkins rod-lens nephroscope has been the standard in percutaneous renal surgery. Despite its high resolution, it is bulky when an external camera and light source are attached. Therefore, new technology to make the scopes lighter and more ergonomic is needed. NEW TECHNOLOGY: The new Invisio Smith digital nephroscope is the first digital nephroscope from Gyrus ACMI. It was used for the first time in two patients with caliceal stones. The tip houses dual LED-driven light carriers and a 1-mm digital camera, which obviate the need for an external light source and camera, thus eliminating the risk of drape fires and patient burns. The Smith nephroscope is much lighter (470 g vs. 939 g) and more ergonomic when compared to the standard rod-lens nephroscope. CONCLUSIONS: The Smith digital nephroscope is much lighter, more ergonomic and safer. Long term use is needed to test its durability. PMID- 18419206 TI - Digital ureteroscopy: the next step. AB - BACKGROUND: Flexible ureteroscopy is used for diagnosing and treating upper urinary tract diseases. Despite technological advances in making flexible ureteroscopes smaller, they suffer from a grainy image. Therefore, new technology with better resolution is needed. NEW TECHNOLOGY: The new Invisio DUR-D digital flexible ureteroscope from Gyrus ACMI was tested. The tip houses dual LED-driven light carriers, which obviates the need for an external light source, thus eliminating the risk of drape fires and patient burns. A 1-mm digital camera at the tip eliminates the need for fragile low-resolution fiberoptics and provides superior resolution. Since there are no external cameras or light cables, the DUR D is much lighter (505 g compared with 1012 g). Laser detection system deactivates the laser to prevent accidental misfiring of the laser within the ureteroscope. CONCLUSIONS: The latest generation of digital ureteroscopes provides superior resolution and safety. Long term use is needed to test its durability. PMID- 18419207 TI - Thoracic complications of urologic laparoscopy: correlation between radiographic findings and clinical manifestations. AB - BACKGROUND AND PURPOSE: The usefulness of plain film chest radiography (CXR) in evaluation for thoracic complications after laparoscopic urologic procedures is uncertain. Our objectives were to examine the association between radiographic findings and clinical manifestations of thoracic complications after laparoscopic urologic procedures and to determine the prevalence of postoperative CXR at our institution. PATIENTS AND METHODS: We performed a retrospective review of 195 patients who underwent laparoscopic renal/adrenal urologic procedures at our institution from 1998 to 2005. Chi-square analysis was used to compare the rate of radiographic abnormalities and thoracic complications between different types of laparoscopic procedures. RESULTS: A total of 96 patients (96/195, 49%) had postoperative CXR, and abnormalities were noted in 75 (75/96, 78%). The abnormalities seen on CXR included atelectasis, pleural effusions, pneumomediastinum, pneumothorax, subcutaneous emphysema, and pneumonia. Retroperitoneal laparoscopy had significantly more incidental subclinical pneumothoraces (P = 0.000469) and subcutaneous emphysema (P = 0.043) identified by CXR than either transperitoneal, hand-assisted, or cryosurgery. Overall, eight patients (8.3%) had clinical manifestations of a thoracic complication but only five (5.2%) were clinically significant complications detected by CXR. Thus, while 75 CXRs were noted as abnormal, 70 (93%) documented incidental findings that did not affect patient care. CONCLUSIONS: Thoracic complications after laparoscopic urologic procedures are uncommon events. Although the majority of CXRs after such procedures do contain abnormalities, most abnormalities are subclinical and do not affect postoperative management. Patients with significant radiographic findings demonstrated significant clinical symptoms. Thus, routine CXR after urologic laparoscopy does not appear to be necessary to identify thoracic complications and may be overused. PMID- 18419208 TI - The role of extracorporeal shockwave lithotripsy in an asymptomatic special patient group with small renal calculi. AB - BACKGROUND AND PURPOSE: Data concerning extracorporeal shock wave lithotripsy (SWL) management of small (< or = 5 mm) asymptomatic renal caliceal stones are lacking in the literature. In this study, we aimed to determine the effectiveness of SWL in a special group of patients who had very small (< or = 5 mm) asymptomatic renal caliceal stones. The group of patients consisted of pilots, gendarmes, and soldiers, such as commandos, for whom treatment was obligatory and who had to be stone free because of their military duties. PATIENTS AND METHODS: We retrospectively examined SWL data and retrieved information for 84 patients with small asymptomatic renal caliceal stones. The SWL sessions were performed with a Siemens Lithostar lithotriptor. The mean age of the patients was 34 +/- 13 years. The mean stone size was 4.8 +/- 0.4 mm. The mean numbers of shockwaves and energy used were 2707 +/- 1742 and 18 +/- 2 kV, respectively. The location of stones were: 11 right upper calix, 10 right middle calix, 17 right lower calix, 13 left upper calix, 16 left middle calyx, and 17 left lower calix. Eighty-four patients underwent 101 SWL sessions. Only eight patients had second and three patients had third SWL treatments. SWL was performed five times for only one patient. All mean values were realized with an SPSS 10.0 statistical program. RESULTS: After the first, second, and third SWL sessions, the stone-free rate was calculated as 87%, 93% and 94%, respectively. Only five patients with lower renal caliceal stones were not stone free. The reported complications were macroscopic hematuria necessitating medical attention in four patients, skin ecchymosis in seven patients, severe colic pain in two patients, and urinary-tract infection in one patient. There was no need for hospitalization for these complications. CONCLUSION: SWL is effective and safe in patients with very small (< or = 5 mm) asymptomatic renal caliceal stones in all locations. PMID- 18419209 TI - Use of LMA Stonebreaker as an intracorporeal lithotrite in the management of ureteral calculi. AB - BACKGROUND AND PURPOSE: LMA Stonebreaker is a new type of ballistic intracorporeal lithotrite that does not need external electric power or access to compressed air. It is small and portable. This study aims to evaluate the efficacy, safety, and cost-effectiveness of this lithotrite in the management of ureteral calculi. MATERIALS AND METHODS: A total of 110 patients with ureteral calculi necessitating intracorporeal lithotripsy were prospectively included in the study. The size of the stone, position of the stone, number of shocks needed to fragment the stone to effect complete clearance, and degree of retropulsion were documented in each case, and any evidence of urothelial trauma was noted. RESULTS: All stones were fragmented, and all patients were rendered stone free. The mean number of shocks needed to fragment the stones was eight. The incidence of retropulsion was 6.36%. There was no evidence of urothelial trauma noted in any patient. CONCLUSION: LMA Stonebreaker is a safe, effective, cost-effective, robust, and portable device for intracorporeal lithotripsy. PMID- 18419210 TI - Cold-knife retrograde endopyelotomy: a long-term follow-up. AB - Cold-knife retrograde endopyelotomy is a simple, safe, and effective therapeutic option for primary and secondary symptomatic ureteropelvic junction obstruction (UPJO). Our long-term success rate with negligible complications demonstrates its durability and safety. Retrograde endopyelotomy should be considered a viable first-line treatment option for the management of patients with UPJOs. PMID- 18419211 TI - Transected guidewire tip simulating ureteral stone. AB - A 62-year-old woman was admitted with a diagnosis of a distal ureteral stone 2 years after antegrade imaging of the kidney. Ureteroscopy revealed a guidewire tip that transected during the antegrade study. The events leading to guidewire transection were reproduced, and a minor modification of current guidewires is suggested to prevent similar incidents. PMID- 18419212 TI - Laparoscopic heminephrectomy in horseshoe kidney using bipolar energy: report of three cases. AB - Variability in blood supply, the renal isthmus, and abnormal kidney location are unique features that contribute to making a laparoscopic approach for heminephrectomy in a horseshoe kidney technically challenging. We report three cases of laparoscopic heminephrectomy for benign disease in horseshoe kidneys using bipolar energy for transection of the isthmus. PMID- 18419213 TI - Antegrade versus retrograde stenting in laparoscopic pyeloplasty. AB - PURPOSE: To compare operative times between retrograde and antegrade ureteral stenting as part of laparoscopic pyeloplasty. PATIENTS AND METHODS: Laparoscopic pyeloplasty procedures from January 2002 to January 2007 were identified through a prospective database. Procedures on 126 patients were performed using the same transperitoneal technique apart from the method of stent placement, which was performed in either a retrograde manner before laparoscopy or an antegrade manner during the laparoscopic portion of the procedure. RESULTS: A total of 45 patients underwent antegrade stenting, 53 had retrograde stenting, 20 patients already had a stent in place, and 8 patients had retrograde pyelography followed by antegrade stenting. Operative time in patients with antegrade stent placement was significantly faster than in those with retrograde stent placement (median 185 v 245 min, P < 0.0001 [two-way analysis of variance]), even when the variability of the operative surgeon was taken into account. There was no difference in the complication rates. CONCLUSION: Antegrade stent placement results in a significantly faster overall operative time when compared with retrograde stent placement. PMID- 18419214 TI - Outcome of retroperitoneoscopic nephrectomy for benign nonfunctioning kidney: a single-center experience. AB - OBJECTIVE: To analyze the feasibility and outcome of retroperitoneoscopic nephrectomy for benign nonfunctioning kidneys and compare it with open simple nephrectomy. MATERIALS AND METHODS: From January 1998 to December 2006, 505 retroperitoneoscopic nephrectomies were performed. In the same time period, 112 open nephrectomies were also performed. In the retroperitoneoscopic group, the mean age was 39 years (range 15-74 years); 204 (40.4%) were men and 301 (59.6%) were women. Forty in this group had a history of surgery. Thirty-six patients had a pyonephrotic kidney; 33 of these patients had undergone percutaneous nephrostomy preoperatively. The cause of the nonfunctioning kidney was ureteropelvic junction obstruction in 198 patients, calculus disease in 193 patients, genitourinary tuberculosis in 48 patients, renal dysplasia in 19 patients, anomalous kidney in 20 patients, and renovascular hypertension in 16 patients. In 11 patients, there were other causes for the nonfunctioning kidney. RESULTS: Retroperitoneoscopic nephrectomy was performed in 476 (94.2%) patients. Conversion to open nephrectomy was necessary in 25 patients. The mean operative time was 85 minutes (range 45-240 min) in the retroperitoneoscopic group and 70 minutes (range 35-120 min) in the open group. The mean blood loss was 110 mL (range 30-600 mL) in the retroperitoneoscopic group and 170 mL (range 70-500 mL) in the open group. Four (0.8%) patients in the retroperitoneoscopic group needed a blood transfusion, whereas 5 (4.5%) patients in the open group had a blood transfusion. The hospital stay in the retroperitoneoscopic group was 3 days (range 1-7 d) and was 5 days (range 3-12 d) in the open group. CONCLUSIONS: Retroperitoneoscopic nephrectomy, although technically challenging, is becoming a gold standard for patients with nonfunctioning kidneys caused by benign conditions. PMID- 18419215 TI - Robot-assisted radical cystectomy in women: technique and initial experience. AB - BACKGROUND AND PURPOSE: Robot-assisted radical cystectomy (RARC) is a new management option for the treatment of bladder cancer. This study evaluates an initial experience with RARC with ileal conduit diversion in women. PATIENTS AND METHODS: Twenty patients underwent RARC with ileal conduit urinary diversion, including four women, and our surgical technique is described here. A retrospective chart review was performed to evaluate clinical stage, tumor grade, operative times, estimated blood loss (EBL), pathologic stage, lymph node pathology, and complications. RESULTS: Mean patient age was 69.5 years, median operative time was 350 minutes, and median EBL was 300 mL. Median length of stay was 5 days, with the two most recent patients leaving by postoperative day 3. The median number of lymph nodes removed was 12, with one patient revealing node positive disease. Surgical margins were negative for disease in all patients. No patients required blood transfusion or had major complications. CONCLUSION: RARC is a new technique available for the treatment of high-risk or invasive bladder cancer in women. This surgery provides decreased morbidity while maintaining the oncologic goals of traditional radical cystectomy. PMID- 18419216 TI - High-energy transurethral thermotherapy with CoreTherm approaches transurethral prostate resection in outcome efficacy: a meta-analysis. AB - BACKGROUND AND PURPOSE: Minimally invasive office-based treatments for benign prostatic hyperplasia (BPH) are challenging the traditional surgical and medical management options for symptomatic BPH. We conducted a meta-analysis of published randomized controlled trials that compared high-energy transurethral microwave thermotherapy (HE-TUMT) with transurethral resection of the prostate (TURP) to compare subjective and objective outcomes. MATERIALS AND METHODS: A literature search using Pub-Med was conducted to obtain all published data on HE-TUMT and all randomized controlled trials that compared HE-TUMT with TURP. Data were analyzed focusing on the pretreatment and posttreatment end points of the International Prostate Symptom Score(IPSS), maximum flow rate (Q(max)), and postvoid residual (PVR). A meta-analysis was conducted, and data were stratified with respect to the type of HE-TUMT machinery used. RESULTS: A total of 458 patients were studied. Differences in IPSS, Q(max), and PVR from current trials that compared TURP with HE-TUMT are best evaluated at 1-year follow-up. At this time point, changes in Q(max) (P < 0.001), IPSS (P = 0.01), and PVR (P = 0.02) are more significant if TURP is the management mode. HE-TUMT with the CoreTherm() device demonstrates the most significant improvements in subjective and objective criteria that approximate outcomes with TURP (Figs. 1-3). CONCLUSIONS: A meta analysis of current randomized controlled trials that compared TURP with HE-TUMT demonstrates more significant changes in Q(max), IPSS, and PVR when TURP is used to manage symptomatic BPH. Despite these statistical differences, stratified data demonstrate that current HE-TUMT machinery is more effective than previously used lower-energy machinery, especially at objective end points. This is most evident when the CoreTherm device is used. These findings, coupled with the decreased costs and morbidity associated with HE-TUMT, support this treatment as a reasonable alternative to TURP. PMID- 18419217 TI - A novel training model for laparoscopic pyeloplasty using chicken crop. AB - PURPOSE: To create a model for laparoscopic pyeloplasty training using the crop and esophagus of a chicken. The model can be used to simulate the steps taken during laparoscopic pyeloplasty and to help trainees practice laparoscopic suturing skills. MATERIALS AND METHODS: The chicken crop and esophagus were used to simulate the renal pelvis and ureter, respectively. These were exposed by reflecting the skin overlying the neck and thorax. The crop was thoroughly cleaned and filled with water via the esophageal end to simulate the dilated renal pelvis. The chicken was positioned within an indigenously made laparoscopic training box. Laparoscopic pyeloplasty was performed using the dismembered Anderson Hynes technique. The model was used over a period of 1 month by three urology trainees in their final year of training. They were assessed with respect to time needed to complete anastomosis and quality of anastomosis. RESULTS: The mean operative time showed a marked reduction from the second to the fourth attempt. There was also a significant improvement in the quality of anastomosis from the first to the fourth attempt. At the end of four attempts, all trainees were able to satisfactorily complete a good quality ureteropelvic anastomosis in a mean time of 67.7 minutes (range 62-76 min). CONCLUSION: Laparoscopic suturing skills require effective training and constant practice to perfect the technique. Adequate practice on this chicken model shortens the learning curve, makes the trainee more confident of his or her skills, and improves his operative performance. PMID- 18419218 TI - Use of free peritoneal and bladder mucosal grafts as ureteral mucosa substitutes for management of avulsion of the ureteral mucosa in a dog model. AB - BACKGROUND AND PURPOSE: Avulsion of the ureteral mucosa is one of the most serious complications of ureteroscopy and considered challenging for urologic surgeons. In this study, we developed two novel methods to reconstruct the impaired ureter. MATERIALS AND METHODS: After ureteral mucosal avulsion in a canine model, ureteral reconstruction was performed in one of three ways: free tabularized peritoneal grafts (group A, n = 6); bladder mucosal grafts (group B, n = 6); or simple stenting technique (control group [n = 3]). At postoperative week 10, evaluation with intravenous urography (IVU) was followed by harvesting of reconstructed ureter segments for pathologic analysis. RESULTS: IVU showed nonvisualization of the collecting system of the affected side and histologic evaluation showed imperforation or stenosis in all animals in the control group. IVU showed good visualization of the reconstructed ureter without stenosis in group A and in most animals in group B. There was a mild dilation but no obvious stenosis in one animal in group B. Microscopically, all the reconstructed ureteral lumens were uniformly lined with transitional epithelium. Extensive neovascularity was evident in the subepithelial layer with a streaming of fibroblasts toward the neomucosa. CONCLUSION: Replacing the avulsed mucosa in the injured ureter with tubularized peritoneal free grafts or bladder mucosal grafts may be an optimal method to manage avulsion of the ureteral mucosa. PMID- 18419219 TI - Efficacy of the sonolith vision lithotripter according to the electrode lifespan: an in vitro study. AB - BACKGROUND AND PURPOSE: Electrodes used by the Sonolith Vision lithotripter have a specified lifespan, and the manufacturer recommends replacing the electrode every 35,000 shocks. To evaluate the efficacy of extracorporeal shockwave lithotripsy (SWL) as it relates to electrode lifespan, and establish standard treatment results, we performed an in vitro study. MATERIALS AND METHODS: From August to October 2005, 12 sessions of shock testing were performed by a Sonolith Vision lithotripter using four test stones per month. The numbers of consumed electrode shocks (CES) at the beginning of testing were classified as: category 1: 0-9999; category 2: 10,000-19,999; category 3: 20,000-29,999; and category 4: 30,000-35,000 shocks. In each category, the number of test stone broken (TBS) shocks was recorded. The voltage was maintained at 25 kV (100%) for every session. The results were analyzed statistically. RESULTS: TBS scores increased with increasing CES. The mean rates of increase in TBS between categories were 14.7% (category 1 to category 2), 8.4% (category 2 to category 3), and 13.7% (category 3 to category 4). There was a strong positive correlation between CES and TBS scores (r = 0.987; P = 0.01). CONCLUSIONS: As the number of consumed electrode shocks increased, the number of test stone broken shocks increased, by an average rate of 12%. Thus when the consumed electrode shocks increase by 10,000 units, the user should increase the number of shocks by about 12% to accomplish equal treatment for all patients. PMID- 18419220 TI - Editorial: innovative approaches in endoscopic surgery. PMID- 18419221 TI - Overcoming obstacles: nerve-sparing issues in radical prostatectomy. AB - PURPOSE: To review, compare, and contrast recovery of potency after robot assisted radical prostatectomy with specific regard to thermal and excisional nerve injury. Our goal was to compare intensity of injury and ability to recover potency in order to stratify risk by injury type. METHODS: We compare potency outcomes in our first 500 consecutive cases of robot-assisted radical prostatectomy. This is a retrospective review of data collected prospectively into an electronic database. Preoperative inclusion criteria were age less than 66 years, International Index of Erectile Function Score of 22-25, and unilateral or bilateral nerve preservation. Potency data were collected via patient-reported validated questionnaires. Potency was defined with two affirmative responses to: "erections hard enough for sexual intercourse" and were they "satisfactory." Results were collated and graphically analyzed so that time-line comparisons of recovery could be evaluated. RESULTS: Time-line curves were generated comparing recovery of sexual function over 2 years after nervesparing techniques using cautery versus cautery-free and unilateral versus bilateral nerve preservation. Use of no cautery improved early return of sexual function 4.7-fold over cautery. Whether using cautery or cautery-free techniques, a doubling of nerve tissue from one nerve to two nerves spared only resulted in a 1.2-fold improvement of potency recovery both early and long-term. CONCLUSIONS: In our experience, avoidance of thermal injury produces nearly a 5-fold improvement in early return of sexual function. Furthermore, thermal injury appears to induce a dense but largely recoverable injury after 2 years. In contrast to common belief, our results demonstrated that preservation of just one nerve will, in the majority of patients, result in similar potency recovery to that with two nerves preserved. Crossover innervation of the one nerve is favored over compensation and hints that techniques that increase nerve volume at the expense of positive surgical margins may need careful introspection. PMID- 18419222 TI - Nephrectomy and natural orifice translumenal endoscopy (NOTES): transvaginal, transgastric, transrectal, and transvesical approaches. AB - This paper outlines recent developments in the field of natural orifice translumenal endosurgery (NOTES) in urology and nephrectomy from the perspective of a flexible endoscopist. The history of transvaginal extraction of kidney specimens using rigid laparoscopic instruments and early studies of transvaginal nephrectomy as a precursor of NOTES are reviewed. Transgastric approaches to nephrectomy using flexible instruments and transvesical hybrid approaches to nephrectomy and other intra-abdominal procedures including cholecystectomy and transvesical lung biopsy are outlined, as are other experimental approaches including transrectal kidney removal. The paper reviews the rationale for a NOTES approach to abdominal surgery and discusses recent human studies. The limitations of NOTES and need for better instrumentation and more studies are stressed. PMID- 18419223 TI - Launching a successful robotic surgery program. AB - Robotic-assisted laparoscopic prostatectomy is rapidly becoming the most commonly performed surgical approach to treat clinically localized prostate cancer. The establishment of a robotic surgery program at any institution requires a structured plan and certain key elements to be in place to allow successful development. At least five essential phases are necessary for successful implementation of a robotics program. A thorough initial design and implementation lead to the execution of clinical services that meet previously established goals. Once the execution phase is established, the next step is to focus on maintenance and growth to maximize the benefits of the program. In this paper, we discuss the necessary phases for creating a successful robotic program, paying special attention to the aspects that allowed our facility to create a profitable robotic-assisted laparoscopic prostatectomy program in year 1. PMID- 18419224 TI - Robotics in urology: past, present, and future. AB - The modern-day urologist is continually armed with new instruments and technology aimed at decreasing the overall invasiveness of urologic procedures. Robotic technology is aimed at improving clinical outcomes by correcting human technical inadequacies such as hand tremors and imprecise suturing. The first reported use of robotics to assist with surgery was in 1985, and the first use of robotics in urology was published in 1989. The currently utilized master-slave system (da Vinci Robotic Platform), Intuitive Surgical, Sunnyvale, CA) has popularized robotic surgery for use in numerous urologic conditions including prostate cancer, bladder cancer, renal cancer, uretero-pelvic junction obstruction, and pelvic prolapse. New developments in robotic technology may revolutionize many other aspects of urology including percutaneous renal access and rounding on patients after surgery. This review provides a brief overview of the history of robotics in urology, a description of the da Vinci surgical system and its current utilization as well as limitations, and a review of evolving robotic technology in the field of urology. PMID- 18419225 TI - Evaluation of a new multi-basket load-release ureteral stent. AB - INTRODUCTION: A multi-basket ureteral stent was tested and modified to assess feasibility of load-release features intended to release stone(s) if resistance is encountered during stent removal. METHODS: Three versions of the stent were evaluated: (1) unaltered stent, (2) one strand of each basket partially cut to 50%, or 75% (3) tube between baskets longitudinally slit. One of the baskets was filled with multiple 3-4-mm stones or a single large 6-mm stone to 20Fr diameter. The force required to release the device through a tube with an 8Fr narrowing was determined. In a separate assessment, urologists used a string attached to the force gauge to exert the maximum force they would be comfortable using in ureteral stent removal. RESULTS: The unaltered stent was associated with higher release forces or inability to remove the stent: 18.67 +/- 8.35 N when a single large stone was present, compared to 8.46 +/- 2.75 N for multiple small stones. Modified devices with 50% and 75% cut basket strands released at 15.19 +/- 1.54 N, and 10.65 +/- 2.09 N, respectively. The longitudinal slit device released at 3.11 +/- 1.57 N. The maximum force exerted by urologists in simulated stent removal was 7.3 +/- 0.4 N. CONCLUSION: Modifications to the multi-basket ureteral stent allow for stent removal in an in vitro model with a narrowing present with stone(s) in the stent baskets. The longitudinal slit modification appears to allow for stent removal with forces low enough to prevent significant ureteral injury, even if ureteral strictures are present. PMID- 18419230 TI - The functions of Deoxyribonuclease II in immunity and development. AB - Apoptosis, which is usually accompanied by DNA degradation, is important not only for the homeostasis of metazoans but also for mammalian development. If DNA is not properly degraded in these processes, it can cause diverse diseases, such as anemia, cataracts, and some autoimmune diseases. A large effort has been made to identify these nucleases that are responsible for these effects. In contrast to Deoxyribonuclease I (DNase I), Deoxyribonuclease II (DNase II) has been less well characterized in these processes. Additionally, enzymes of DNase II family in Trichinella spiralis, which is an intracellular parasitic nematode, are also considered involved in the development of the nematode. We have compiled information from studies on DNase II from various organisms and found some nonclassic features in these enzymes of T. spiralis. Here we have reviewed the characterization and functions of DNase II in these processes and predicted the functions of these enzymes in T. spiralis during host invasion and development. PMID- 18419231 TI - Cell-nanofiber-based cartilage tissue engineering using improved cell seeding, growth factor, and bioreactor technologies. AB - Biodegradable nanofibrous scaffolds serving as an extracellular matrix substitute have been shown to be applicable for cartilage tissue engineering. However, a key challenge in using nanofibrous scaffolds for tissue engineering is that the small pore size limits the infiltration of cells, which may result in uneven cell distribution throughout the scaffold. This study describes an effective method of chondrocyte loading into nanofibrous scaffolds, which combines cell seeding, mixing, and centrifugation to form homogeneous, packed cell-nanofiber composites (CNCs). When the effects of different growth factors are compared, CNCs cultured in medium containing a combination of insulin-like growth factor-1 and transforming growth factor-beta1 express the highest mRNA levels of collagen type II and aggrecan. Radiolabeling analyses confirm the effect on collagen and sulfated-glycosaminoglycans (sGAG) production. Histology reveals chondrocytes with typical morphology embedded in lacuna-like space throughout the entire structure of the CNC. Upon culturing using a rotary wall vessel bioreactor, CNCs develop into a smooth, glossy cartilage-like tissue, compared to a rough-surface tissue when maintained in a static environment. Bioreactor-grown cartilage constructs produce more total collagen and sGAG, resulting in greater gain in net tissue weight, as well as express cartilage-associated genes, including collagen types II and IX, cartilage oligomeric matrix protein, and aggrecan. In addition, dynamic culture enhances the mechanical property of the engineered cartilage. Taken together, these results indicate the applicability of nanofibrous scaffolds, combined with efficient cell loading and bioreactor technology, for cell-based cartilage tissue engineering. PMID- 18419233 TI - Prospective predictors of suicide attempts in borderline personality disorder at one, two, and two-to-five year follow-up. AB - Efforts to identify reliable predictors of suicidal behavior in Borderline Personality Disorder have been confounded by the marked dimensional heterogeneity of the disorder, frequent comorbidity with other high risk disorders, debilitating social and vocational consequences of BPD over time. Using survival analyses, we assessed the predictive association between risk factors in each of these symptom domains and suicide attempts in BPD subjects followed for 12 months, 18-24 months and 2-5 years. The suicide attempt rate was 19% in the first year, 24.8% through the second year. The risk of suicidal behavior among 137 BPD subjects completing the first 12 months was increased by comorbid MDD and poor social adjustment. Outpatient treatment decreased short-term risk. Among 133 subjects completing 18-24 months in the study, the relative risk of a suicide attempt was increased by hospitalization (prior to any attempt), and poor social adjustment. Among 122 subjects followed for 2-5 years, increased risk was associated with hospitalization and medication visits (prior to any attempt), an attempt in the first year, and a low GAS score at baseline. Long term risk was decreased by "any outpatient treatment." Predictors of suicidal behavior in BPD change over time. MDD has a short-term effect on suicide risk, while poor social adjustment may increase risk throughout each follow-up interval. Assessing and supporting family, work, and social relationships may decrease suicidal behavior in BPD, and should be a principal focus of long-term treatment. PMID- 18419234 TI - Recognition of facial affect in Borderline Personality Disorder. AB - Patients with Borderline Personality Disorder (BPD) have been described as emotionally hyperresponsive, especially to anger and fear in social contexts. The aim was to investigate whether BPD patients are more sensitive but less accurate in terms of basic emotion recognition, and show a bias towards perceiving anger and fear when evaluating ambiguous facial expressions. Twenty-five women with BPD were compared with healthy controls on two different facial emotion recognition tasks. The first task allowed the assessment of the subjective detection threshold as well as the number of evaluation errors on six basic emotions. The second task assessed a response bias to blends of basic emotions. BPD patients showed no general deficit on the affect recognition task, but did show enhanced learning over the course of the experiment. For ambiguous emotional stimuli, we found a bias towards the perception of anger in the BPD patients but not towards fear. BPD patients are accurate in perceiving facial emotions, and are probably more sensitive to familiar facial expressions. They show a bias towards perceiving anger, when socio-affective cues are ambiguous. Interpersonal training should focus on the differentiation of ambiguous emotion in order to reduce a biased appraisal of others. PMID- 18419235 TI - Impulsivity and emotion dysregulation in Borderline Personality Disorder. AB - This study examined the association of borderline personality disorder (BPD) and negative emotional states with impulsivity in the laboratory. Undergraduate participants who were high in BPD features (high-BPD; n = 39) and controls who were low in BPD features (low-BPD; n = 56) completed measures of negative emotional state before a laboratory measure of impulsivity--a passive avoidance learning task. Controlling for psychopathology, high-BPD participants committed a greater number of impulsive responses than did low-BPD participants. Negative emotional state moderated the effect of BPD on impulsive responses. High-BPD participants who were in a negative emotional state committed fewer impulsive responses than high-BPD participants who were low in negative emotional state. Fear, nervousness, and shame negatively correlated with impulsivity among high BPD participants but not among low-BPD participants. In addition, high-BPD participants reported greater emotion dysregulation in a variety of domains, compared with low-BPD participants. PMID- 18419236 TI - Dysfunctional beliefs and psychopathology in Borderline Personality Disorder. AB - This study examined the factor structure of the Borderline Personality Disorder subscale of the Personality Beliefs Questionnaire (PBQ-BPD; Butler, Brown, Beck, & Grisham, 2002), and the relationships between the emergent factors and psychopathology. The sample comprised 184 patients diagnosed with borderline personality disorder (BPD). Exploratory factor analysis yielded three factors relating respectively to dependency, distrust, and the belief that one should act preemptively to avoid threat. Although the three factors were significantly associated with depression, only dependency and distrust significantly correlated with hopelessness. Distrust was the sole factor that correlated significantly with suicide ideation. These findings support the dimensional structure of the PBQ-BPD. Given its multidimensional structure, the scale can be used as a measure of belief profiles associated with BPD and as an aid to conceptualizing beliefs underlying a range of psychopathology associated with patients with BPD. PMID- 18419237 TI - Discriminant validity of the ZKPQ in a sample meeting BPD diagnosis vs. normal range controls. AB - Widiger and Simonsen (2005) state that given the limitations of the categorical model of Personality Disorders classification proposals are to be expected for dimensional classifications. The purpose of this paper is to test the alternative five factorial model (AFFM) of personality in a sample with PDs. Subjects were administered the ZKPQ to test the discriminant capacity of the AFFM in classifying subjects diagnosed with BPD (n = 74) vs normal-range controls (n = 148) paired by age and sex, and identifying sensitive and/or specific dimensions that can be of help in diagnosing BPD. The results showed that high scores on N Anx and Imp-SS, and low scores on Act are prognostic factors for being diagnosed with BPD. Likewise, this model correctly classified 88% of subjects with a kappa index of 0.73. The AFFM of personality appears to have a substantial power for predicting SCID-II interview-based BPD diagnosis. PMID- 18419238 TI - Personality disorders and pathological gambling: a review and re-examination of prevalence rates. AB - The current study reviews and reexamines the association between pathological gambling and personality disorders (PDs). To date, the majority of investigations have examined the prevalence of PDs in a single group of treatment-seeking pathological gamblers (PGs); very few of these studies included a comparison group, and even fewer compared PGs to nonpathological gamblers who, in contrast to nongamblers, resemble PGs in their attraction to and engagement in gambling behavior. The current study included a sample composed of nontreatment-seeking pathological gamblers and a comparison group of nonpathological gamblers (NPGs); these participants completed a self-report instrument (SCID-II/PQ) and were administered a structured clinical interview SCID-II) designed to assess PDs. Compared to the SCID-II, the SCIDII/PQ produced significantly higher PD prevalence rate estimates and symptom endorsements. Although the pattern of specific PD prevalence and symptom endorsement varied somewhat across the instruments, PGs consistently displayed significantly higher levels of borderline PD than NPGs; this pattern endured even after controlling for Axis I disorders and overlap among Axis II PDs. PMID- 18419239 TI - The association of affiliation and achievement personality styles with DSM-IV personality disorders. AB - Investigators from theoretically diverse backgrounds have long described two personality styles associated with depression. The affiliation style describes individuals with interpersonal concerns; the achievement style describes individuals who have concerns with personal failure. Although primarily related to risk for depression, there has also been recognition that extreme forms of these styles are related to personality disorders (PDs). The current study examines the relation between the DSM-IV PDs and the two personality styles, assessing the latter using two approaches: (a) dependency/self-criticism (Blatt, 1974) and (b) sociotropy/autonomy (Beck, 1983). Results show that whereas the achievement style is associated with most PDs (except for the histrionic and dependent PDs), the affiliation style is associated only with histrionic, dependent, and depressive PDs. Controlling for shared PD variance leaves only two associations, affiliation with dependent PD and achievement with narcissistic PD. Results are discussed in the context of current efforts to capture personality psychopathology within a comprehensive dimensional framework. PMID- 18419240 TI - The prevalence of personality disorders in a Kenyan inpatient sample. AB - DSM-IV Axis I and II comorbidities and the pattern of Axis II diagnoses in patients admitted at Mathari (Psychiatric) Hospital, Nairobi, Kenya are unknown. To determine DSM-IV Axis I and II comorbidities and patterns of Axis II diagnoses in patients admitted at Mathari Hospital. Cross-sectional study on 148 randomly sampled patients. Twenty percent of the patients were confirmed for an Axis II diagnosis. Eighty-seven percent of the Axis II disorders were Cluster B Personality Disorders of various types. Using chi2 tests, significant associations were found between Axis I and II diagnoses and substance use/dependence (p < 0.001; 66.7%), mood disorder (p = 0.002; 46.7%) and schizophrenia (p < 0.001; 23.3%). The prevalence of personality disorders was lower than that reported in psychiatric patients in USA and Europe. The results are likely to be a true reflection of the actual epidemiological situation, but cannot be generalized to outpatient or general populations. PMID- 18419241 TI - Threats of violence to third parties in group psychotherapy. AB - This article considers threats of violence toward third parties during group therapy. An approach to preserve the group work, protect the threatened individuals, maintain the threatening patient's treatment, and protect the group leader medicolegally is described. Obtaining expert legal advice is very important. Reluctance to seek legal advice and inform third parties needs to be resolved. Knowledge of jurisdictional laws pertaining to disclosure is essential. PMID- 18419242 TI - When children tell stories: developmental considerations. AB - Although psychoanalysts welcome children's story creations in the clinical setting, psychoanalytic investigators have given them limited attention. This author examined stories that 40 children, aged 8 to 14, narrated to the Thematic Apperception Test blank card presented in psychological evaluation. The stories depicted playful use of reality as a new development; specific dynamic factors influencing thematic content; humor in latency and self-referencing in (pre )adolescence as age-specific characteristics. The findings further our understanding of conscious fantasy. PMID- 18419243 TI - A continuing medical education approach to improve sexual boundaries of physicians. AB - INTRODUCTION: Physician sexual boundary violations are a public health problem. Few resources exist to address physicians who behave inappropriately with patients. In response, the Center for Professional Health at Vanderbilt University developed a three-day continuing medical education (CME) course about proper professional sexual boundaries in 2000. The mission of this CME course is to offer an educational intervention for those physicians whose professional sexual misconduct has required such education as part of a larger accountability sanction. Previous studies suggest that when such education is offered through non-traditional medical education, it is effective in promoting behavioral change. This paper describes the three-day intensive educational experience offered by a CME course with a particular focus on lessons learned from more than 7 years of experience working with these physicians. METHODS: Over 381 physicians from 40 states and Canada have attended. Data about course participants was collected by self-report and aggregated into three categories: demographics, results of assessment tools administered, and quality of the experience. Assessment tools used include the Family Adaptability and Cohesion Evaluation Scale II (FACES II), the Trauma Symptom Inventory (TSI) and the Sexual Addiction Screening Test (SAST). RESULTS: Most physicians were referred to the course from physician health programs and boards of medical examiners. The majority of physician participants were male and in group or solo practice. A full range of medical specialties was represented with most physicians being internists, psychiatrists, obstetricians and surgeons. Results of assessment tools administered indicate that physicians referred for sexual boundary violations often come from dysfunctional families and demonstrate symptoms indicative of trauma related problems and possible sexual addiction. Physician attendees report being highly satisfied with the new knowledge attained in this course. DISCUSSION: Curriculum aimed at addressing sexual boundary violations should address family of origin issues, trauma coping skills and sexual acting out. Satisfaction data continues to support a small group, experiential, and confidential format as an effective means for intervention. CONCLUSION: A CME course offers a model for future training experiences for faculty, residents, medical students and community physicians to teach skills that may help prevent and remediate professional boundary crossings. PMID- 18419244 TI - Rorschach correlates of adolescent self-mutilation. AB - The present study examined various Rorschach variables of aggression, dysphoric affect, and disordered thinking in relation to self-mutilation in samples of self mutilating (SM; n=16) and nonself-mutilating (N-SM; n=26) adolescent inpatients. Categorical comparisons indicated that SM patients had significantly higher mean scores for Aggressive Past (AgPast), but not for Aggressive Content (AgC), Aggressive Movement (AG), Morbid Content (MOR), Inanimate Movement (m), Sum Shading (SumY), and the Perceptual-Thinking Index (PTI), than N-SM patients. Additionally, logistic regression results suggested that PTI and AgPast were the most robust predictors of group membership. The authors propose that self mutilating adolescents may struggle with internally directed aggression and victimized sense of self that, when coupled with disordered thinking, significantly predicts self-mutilation. PMID- 18419246 TI - Extramedullary chitosan channels promote survival of transplanted neural stem and progenitor cells and create a tissue bridge after complete spinal cord transection. AB - Transplantation of neural stem and progenitor cells (NSPCs) is a promising strategy for repair after spinal cord injury. However, the epicenter of the severely damaged spinal cord is a hostile environment that results in poor survival of the transplanted NSPCs. We examined implantation of extramedullary chitosan channels seeded with NSPCs derived from transgenic green fluorescent protein (GFP) rats after spinal cord transection (SCT). At 14 weeks, we assessed the survival, maturation, and functional results using NSPCs harvested from the brain (brain group) or spinal cord (SC group) and seeded into chitosan channels implanted between the cord stumps after complete SCT. Control SCT animals had empty chitosan channels or no channels implanted. Channels seeded with brain or spinal cord-derived NSPCs showed a tissue bridge, although the bridges were thicker in the brain group. Both cell types showed long-term survival, but the number of surviving cells in the brain group was approximately five times as great as in the SC group. In both the brain and SC groups at 14 weeks after transplantation, many host axons were present in the center of the bridge in association with the transplanted cells. At 14 weeks astrocytic and oligodendrocytic differentiation in the channels was 24.8% and 17.3%, respectively, in the brain group, and 31.8% and 9.7%, respectively, in the SC group. The channels caused minimal tissue reaction in the adjacent spinal cord. There was no improvement in locomotor function. Thus, implantation of chitosan channels seeded with NSPCs after SCT created a tissue bridge containing many surviving transplanted cells and host axons, although there was no functional improvement. PMID- 18419249 TI - Trichostatin A improves histone acetylation in bovine somatic cell nuclear transfer early embryos. AB - Epigenetic aberrancies likely preclude correct and complete nuclear reprogramming following somatic cell nuclear transfer (SCNT), and may underlie the observed reduced viability of cloned embryos. In the present study, we tested the effects of the histone deacetylase inhibitor (HDACi), trichostatin A (TSA), on development and histone acetylation of cloned bovine preimplantation embryos. Our results indicated that treating activated reconstructed SCNT embryos with 50 nM TSA for 13 h produced eight-cell embryos with levels of acetylation of histone H4 at lysine 5 (AcH4K5) similar to fertilized counterparts and significantly greater than in control NT embryos (p < 0.005). Further, TSA treatment resulted in SCNT embryos with preimplantation developmental potential similar to fertilized counterparts, as no difference was observed in cleavage and blastocyst rates or in blastocyst total cell number (p > 0.05). Measurement of eight selected developmentally important genes in single blastocysts showed a similar expression profile among the three treatment groups, with the exception of Nanog, Cdx2, and DNMT3b, whose expression levels were higher in TSA-treated NT than in in vitro fertilized (IVF) embryos. Data presented herein demonstrate that TSA can improve at least one epigenetic mark in early cloned bovine embryos. However, evaluation of development to full-term is necessary to ascertain whether this effect reflects a true increase in developmental potential. PMID- 18419250 TI - Androgen regulates neuritin mRNA levels in an in vivo model of steroid-enhanced peripheral nerve regeneration. AB - Following crush injury to the facial nerve in Syrian hamsters, treatment with androgens enhances axonal regeneration rates and decreases time to recovery. It has been demonstrated in vitro that the ability of androgen to enhance neurite outgrowth in motoneurons is dependent on neuritin-a protein that is involved in the re-establisment of neuronal connectivity following traumatic damage to the central nervous system and that is under the control of several neurotrophic and neuroregenerative factors--and we have hypothesized that neuritin is a mediator of the ability of androgen to increase peripheral nerve regeneration rates in vivo. Testosterone treatment of facial nerve-axotomized hamsters resulted in an approximately 300% increase in neuritin mRNA levels 2 days post-injury. Simultaneous treatment with flutamide, an androgen receptor blocker that is known to prevent androgen enhancement of nerve regeneration, abolished the ability of testosterone to upregulate neuritin mRNA levels. In a corroborative in vitro experiment, the androgen dihydrotestosterone induced an approximately 100% increase in neuritin mRNA levels in motoneuron-neuroblastoma cells transfected with androgen receptors, but not in cells without androgen receptors. These data confirm that neuritin is under the control of androgens, and suggest that neuritin is an important effector of androgen in enhancing peripheral nerve regeneration following injury. Given that neuritin has now been shown to be involved in responses to both central and peripheral injuries, and appears to be a common effector molecule for several neurotrophic and neurotherapeutic agents, understanding the neuritin pathway is an important goal for the clinical management of traumatic nervous system injuries. PMID- 18419251 TI - Changes in corticospinal function and ankle motor control during recovery from incomplete spinal cord injury. AB - Little is known about the mechanisms that underlie motor recovery after incomplete spinal cord injury (iSCI) in humans. This study assessed changes in corticospinal tract (CST) function by measuring motor-evoked potentials (MEPs) and ankle motor control at 1, 3, and 6 months after acute iSCI. In 12 iSCI patients and matched controls, MEPs (evoked at 20% of maximal voluntary contraction [MVC]) were combined with a comprehensive ankle motor assessment protocol that measured ankle dorsiflexor strength (MVC, manual muscle testing, maximal movement velocity [MMV]), dexterity (the ability to accurately time ankle dorsiflexion movements) and gait (speed, walking aids). In the first 6 months after iSCI, all measures of muscle strength, gait and the MEP amplitudes significantly increased. The level of background electromyography (EMG) at 20% MVC remained stable, although absolute MVC increased. The MEP latencies were significantly delayed and remained unchanged during the first 6 months after iSCI. In addition, dexterity was preserved throughout rehabilitation. The percentage increase in MEP amplitude was significantly related only to the percentage improvement in MMV. The finding of unchanged CST conductivity, as assessed by MEP latencies in acute iSCI patients recovering motor function, is in accordance with previous studies in human SCI on this issue. The increased MEP facilitation at stable background EMG might indicate improved synchronization of the descending volley and/or responsiveness of motoneurons to supra-spinal input. The absence of a relationship between MEP amplitudes and recovery of ambulation and muscle strength implies that plastic changes in spinal neural circuits and preserved motor units might have contributed to the functional improvement. PMID- 18419252 TI - Time of hypotension and discharge outcome in children with severe traumatic brain injury. AB - We performed a retrospective study at a level I pediatric trauma center of patients admitted between 1998 and 2005 to determine the time after severe pediatric traumatic brain injury (TBI) that hypotension (systolic blood pressure [SBP] of <5th percentile) is most strongly associated with poor outcome. One hundred forty-six patients of <18 years of age with TBI, head Abbreviated Injury Score (AIS) of >or=3, and PICU admission Glasgow Coma Scale (GCS) score of <9 formed the analytic sample. Available SBP readings through the first 72 h after severe TBI were collected. SBP of <5th percentile was defined as hypotension. Discharge Glasgow Outcome Scale (GOS) score of <4 defined poor outcome. Of 146, 59 (40%) patients had discharge GOS of <4 and 12% died. The adjusted risk of poor outcome associated with hypotension stabilized by 8 h (adjusted risk ratio [RR] 1.7; 95% confidence interval [CI] 1.1-2.6) after injury. The risk of poor outcome peaked with hypotension occurring within the first 6 h after injury (RR 2.0, 95% CI 1.3-3.3). Poor discharge GOS was predicted by hypotension occurring during the first 6 h after injury. SBP data beyond the first 6 h did not improve our ability to predict poor discharge GOS. The first 6 h after severe pediatric TBI may represent a critical time period for either predicting or improving outcome. PMID- 18419253 TI - Regulation of immunity to respiratory syncytial virus by dendritic cells, toll like receptors, and notch. AB - The activation and maintenance of pulmonary viral disease is regulated at multiple levels and determined by the early innate response to the pathogenic stimuli. Subsequent activation events that rely directly and indirectly on the virus itself can alter the development and severity of the ensuing immunopathologic responses. In the present review we outline several interconnected mechanisms that rely on the early recognition of viral nucleic acid for the most appropriate anti-viral immune responses, including TLRs and Notch activation in DCs and T cells. Deviation or persistence of the immune response to respiratory viruses may impact significantly on the severity of the responses. While these mechanisms are likely similar in most respiratory viral infections, this review will focus on findings with respiratory syncytial virus (RSV) infections. PMID- 18419255 TI - Intensity of platelet beta(3) integrin in patients with hemorrhagic fever with renal syndrome and its correlation with disease severity. AB - beta(3) Integrin has been identified as a cellular receptor for Hantaan virus, which causes hemorrhagic fever with renal syndrome (HFRS). To investigate the relationship between intensity of the platelet membrane beta(3) integrin (CD61) and disease severity, the percentage of CD61-positive platelets and the mean fluorescence intensities (MFI) of platelet CD61 were determined in patients with HFRS by flow cytometry. The intensity levels of CD61 in patients with HFRS were significantly higher than those in the controls and correlated with the clinical phases of the disease. The CD61 intensity at the oliguric phase was inversely correlated with platelet count and serum albumin, and positively correlated with white blood cell count, blood urea nitrogen, serum creatinine, and alanine aminotransferase levels. The results suggest that the intensity levels of platelet CD61 were elevated and associated with clinical phases and disease severity in patients with HFRS, and the intensity of platelet beta(3) integrin in patients with HFRS may be indicative of disease severity. PMID- 18419254 TI - Effects of interleukin-12 and interleukin-15 on measles-specific T-cell responses in vaccinated infants. AB - Understanding the infant host response to measles vaccination is important because of their increased mortality from measles and the need to provide effective protection during the first year of life. Measles-specific T and B-cell responses are lower in infants after measles vaccination than in adults. To define potential mechanisms, we investigated age-related differences in measles specific T-cell proliferation, CD40-L expression, and IFN-gamma production after measles immunization, and the effects of rhIL-12 and rhIL-15 on these responses. Measles-specific T-cell proliferation and mean IFN-gamma release from infant PBMCs were significantly lower when compared with responses of vaccinated children and adults. Infant responses increased to ranges observed in children and adults when both rhIL-12 and rhIL-15 were added to PBMC cultures. Furthermore, a significant rise in T-cell proliferation and IFN-gamma release was observed when infant PBMCs were stimulated with measles antigen in the presence of rhIL-12 and rhIL-15 compared to measles antigen alone. CD40-L expression by infant and adult T cells stimulated with measles antigen was comparable, but fewer infant CD40-L(+) T cells expressed IFN-gamma. These observations suggest that lower measles-specific T-cell immune responses elicited by measles vaccine in infants may be due to diminished levels of key cytokines. PMID- 18419256 TI - Identification of human CD4 T-cell epitopes on the VP1 capsid protein of enterovirus 71. AB - The identification of human CD4 T-cell epitopes within a protein vaccine candidate is of great interest,as it provides a better understanding of the mechanisms involved in protective immunity and may therefore help in the design of effective vaccines and diagnostic tools. The entire amino acid sequence of the VP1 capsid protein from enterovirus 71 (EV 71) strain 41 was submitted to analysis by the ProPred algorithm for the identification of potential promiscuous human CD4 T-cellepitopes. Three regions spanning amino acids 66-77, 145-159, and 247-261 of VP1 were predicted to bind more than 25 HLA-DR alleles. The corresponding synthetic peptides (SP1 to SP3) were then tested for their abilities to induce proliferation of CD4 T cells isolated from five human volunteers screened positive for previous EV 71 exposure and one EV 71-negative volunteer. Upon stimulation with either peptide, CD4 T-cell proliferative responses were observed for all EV 71-positive volunteers,indicating the presence of EV 71-specific memory CD4 T cells. The amplitude of the proliferative responses was peptide- and HLA-DR-dependent, and correlated well with the ProPredpredicted binding efficiencies. Moreover, CD4 T cells from EV 71-positive volunteers produced significant levels of IL-2 and IFN- upon stimulation, indicative of a T-cell differentiation into Th-1-type subset. Among the three peptides, SP2 induced the highest proliferative response and cytokine production. Moreover, SP2-induced proliferative response could be inhibited with anti-major histocompatibility complex (MHC) class II antibody, indicating that SP2 represents a MHC class II-restricted CD4 T-cell epitope. This study demonstrates that the ProPred algorithm can accurately predict the presence of human CD4 T cell epitopes within the VP1 capsid protein of EV 71, and therefore represents a useful tool for the design of subunit vaccines against EV 71. PMID- 18419258 TI - Biological Rhythms Workshop I: introduction to chronobiology. AB - In this chapter, we present a series of four articles derived from a Introductory Workshop on Biological Rhythms presented at the 72nd Annual Cold Spring Harbor Symposium on Quantitative Biology: Clocks and Rhythms. A diverse range of species, from cyanobacteria to humans, evolved endogenous biological clocks that allow for the anticipation of daily variations in light and temperature. The ability to anticipate environmental variation promotes optimal performance and survival. In the first article, Introduction to Chronobiology, we present a brief historical timeline of how circadian concepts and terminology have emerged since the early observation of daily leaf movement in plants made by an astronomer in the 1700s. Workshop Part IA provides an overview of the molecular basis for rhythms generation in several key model organisms, Workshop Part IB focuses on how biology built a brain clock capable of coordinating the daily timing of essential brain and physiological processes, and Workshop Part IC gives key insight into how researchers study sleep and rhythms in humans. PMID- 18419259 TI - Biological Rhythms Workshop IA: molecular basis of rhythms generation. AB - Current circadian models are based on genetic, biochemical, and structural data that, when combined, provide a comprehensive picture of the molecular basis for rhythms generation. These models describe three basic elements-input pathways, oscillator, and output pathways-to which each molecular component is assigned. The lines between these elements are often blurred because some proteins function in more than one element of the circadian system. The end result of these molecular oscillations is the same in each system (near 24-hour timing), yet the proteins involved, the interactions among those proteins, and the regulatory feedback loops differ. Here, the currentmodels for the molecular basis for rhythms generation are described for the prokaryotic cyanobacterium Synechococcus elongatus as well as the eukaryotic systems Neurospora crassa, Drosophila melanogaster, Arabidopsis thaliana, and mammals (particularly rodents). PMID- 18419260 TI - Biological Rhythms Workshop IB: neurophysiology of SCN pacemaker function. AB - Pacemakers are functional units capable of generating oscillations that synchronize downstream rhythms. In mammals, the suprachiasmatic nucleus (SCN) of the hypothalamus is a circadian pacemaker composed of individual neurons that intrinsically express a near 24-hour rhythm in gene expression. Rhythmic gene expression is tightly coupled to a rhythm in spontaneous firing rate via intrinsic daily regulation of potassium current. Recent progress in the field indicates that SCN pacemaking is a specialized property that emerges from intrinsic features of single cells, structural connectivity among cells, and activity dynamics within the SCN. The focus of this chapter is on how Nature built a functional pacemaker from many individual oscillators that is capable of coordinating the daily timing of essential brain and physiological processes. PMID- 18419261 TI - Biological Rhythms Workshop IC: sleep and rhythms. AB - Rhythms of sleep and wakefulness (typically measured as rest/activity rhythms) are among the most prominent of biological rhythms and therefore were among the first to be recorded in early chronobiological studies. These rhythms can provide useful information about the central biological clock, although an appreciation of the problems associated with using rest/activity to infer central clock function is important in the design and interpretation of chronobiological experiments in both animals and humans. Here, we review the anatomical and neurophysiologic bases of sleep regulation in mammals as well as similarities and differences between the sleep of humans and that of other organisms. We outline how human sleep is measured, the role of the circadian system in models of human sleep regulation, and human circadian rhythm sleep disorders. Although the function of sleep is still not completely understood, sleep has a critical role for human health, and we have attempted to outline the role that the circadian timing system has in regulating human sleep and in contributing to sleep disorders. PMID- 18419262 TI - A cyanobacterial circadian clock based on the Kai oscillator. AB - In the cyanobacterium Synechococcus elongatus PCC 7942, the products of three genes (kaiA, kaiB, and kaiC) have been identified as essential components of the circadian clock. Recently, we reconstituted the self-sustainable circadian oscillation of the KaiC phosphorylation state by incubating purified KaiC with KaiA, KaiB, and ATP. This in vitro oscillation persisted for at least three cycles and the period was compensated against temperature changes. Period lengths observed in vivo in various kaiC mutants were consistent with those measured using in vitro mixtures containing the respective mutant KaiC proteins. These results demonstrate that the oscillation of KaiC phosphorylation is the primary pacemaker of the cyanobacterial circadian clock and reveal a novel function of proteins as timing devices that govern cellular metabolism. We further analyzed four aspects of the KaiC phosphorylation cycle in vitro: the interactions among KaiA, KaiB, and KaiC; the functions of the two phosphorylation sites, the energetics that determine the circadian period, and the mechanisms that synchronize the components of the Kai oscillator. From these analyses, we have proposed a circadian program consisting of the three proteins that keeps biological time in a living cell. PMID- 18419263 TI - A PER/TIM/DBT interval timer for Drosophila's circadian clock. AB - Circadian rhythms in Drosophila are supported by a negative feedback loop, in which PERIOD (PER) and Timeless (TIM) shut down their own transcription as they translocate once a day from the cytoplasm of clock-containing cells to the nucleus. Period length is partially determined by an interval of cytoplasmic retention of the TIM and PER proteins. To study this process, we examined PER/TIM/Doubletime (DBT) physical interactions and nuclear translocation by imaging individual cultured Drosophila cells. Using live cell video microscopy and green fluorescent protein (GFP) tags, we observed dynamic patterns of stability and localization for DBT, PER, and TIM that resembled those previously found in vivo. These studies suggest that a cytoplasmic interval timer regulates nuclear translocation of these proteins. The cultured cell assay provides a potent system to study interactions among new and known genes involved in the generation of circadian behavior. PMID- 18419264 TI - Transcriptional feedback and definition of the circadian pacemaker in Drosophila and animals. AB - The modern era of Drosophila circadian rhythms began with the landmark Benzer and Konopka paper and its definition of the period gene. The recombinant DNA revolution then led to the cloning and sequencing of this gene. This work did not result in a coherent view of circadian rhythm biochemistry, but experiments eventually gave rise to a transcription-centric view of circadian rhythm generation. Although these circadian transcription-translation feedback loops are still important, their contribution to core timekeeping is under challenge. Indeed, kinases and posttranslational regulation may be more important, based in part on recent in vitro work from cyanobacteria. In addition, kinase mutants or suspected kinase substrate mutants have unusually large period effects in Drosophila. This chapter discusses our recent experiments, which indicate that circadian transcription does indeed contribute to period determination in this system. We propose that cyanobacteria and animal clocks reflect two independent origins of circadian rhythms. PMID- 18419265 TI - Genetic and molecular analysis of the central and peripheral circadian clockwork of mice. AB - A hierarchy of interacting, tissue-based clocks controls circadian physiology and behavior in mammals. Preeminent are the suprachiasmatic nuclei (SCN): central hypothalamic pacemakers synchronized to solar time via retinal afferents and in turn responsible for internal synchronization of other clocks present in major organ systems. The SCN and peripheral clocks share essentially the same cellular timing mechanism. This consists of autoregulatory transcriptional/posttranslational feedback loops in which the Period (Per) and Cryptochrome (Cry) "clock" genes are negatively regulated by their protein products. Here, we review recent studies directed at understanding the molecular and cellular bases to the mammalian clock. At the cellular level, we demonstrate the role of F-box protein Fbxl3 (characterized by the afterhours mutation) in directing the proteasomal degradation of Cry and thereby controlling negative feedback and circadian period of the molecular loops. Within SCN neural circuitry, we describe how neuropeptidergic signaling by VIP synchronizes and sustains the cellular clocks. At the hypothalamic level, signaling via a different SCN neuropeptide, prokineticin, is not required for pacemaking but is necessary for control of circadian behavior. Finally, we consider how metabolic pathways are coordinated in time, focusing on liver function and the role of glucocorticoid signals in driving the circadian transcriptome and proteome. PMID- 18419266 TI - The multiple facets of Per2. AB - The Period 2 (Per2) gene is an important component of the circadian system. It appears to be not only part of the core oscillator mechanism, but also part of the input and output pathways of the clock. Because of its involvement at multiple levels of the circadian system, Per2 needs to meet a variety of different demands. We discuss how Per2 might be able to fulfill multiple functions by reviewing known facts and combine this with speculations based on these facts. This might provide new views about Per2 function and help to better understand diseases that are rooted in the circadian system. PMID- 18419267 TI - Chromatin remodeling and circadian control: master regulator CLOCK is an enzyme. AB - The molecular machinery that governs circadian rhythmicity is based on clock gene products organized in regulatory feedback loops. Recently, we have shown that CLOCK, a master circadian regulator, has histone acetyltransferase activity essential for clock gene expression. The Lys-14 residue of histone H3 is a preferential target of CLOCK-mediated acetylation. As the role of chromatin remodeling in eukaryotic transcription is well recognized, this finding identified unforeseen links between histone acetylation and cellular physiology. Indeed, we have shown that the enzymatic function of CLOCK drives circadian control. We reasoned that CLOCK's acetyltransferase activity could also target nonhistone proteins, a feature displayed by other HATs. Indeed, CLOCK also acetylates a nonhistone substrate: its own partner, BMAL1. This protein undergoes rhythmic acetylation in the mouse liver, with a timing that parallels the down regulation of circadian transcription of clock-controlled genes. BMAL1 is specifically acetylated on a unique, highly conserved Lys-537 residue. This acetylation facilitates recruitment of the repressor CRY1 to BMAL1, indicating that CLOCK may intervene in negative circadian regulation. Our findings reveal that the enzymatic interplay between two clock core components is crucial for the circadian machinery. PMID- 18419268 TI - The ancestral circadian clock of monarch butterflies: role in time-compensated sun compass orientation. AB - The circadian clock has a vital role in monarch butterfly (Danaus plexippus) migration by providing the timing component of time-compensated sun compass orientation, which contributes to navigation to the overwintering grounds. The location of circadian clock cells in monarch brain has been identified in the dorsolateral protocerebrum (pars lateralis); these cells express PERIOD, TIMELESS, and a Drosophila-like cryptochrome designated CRY1. Monarch butterflies, like all other nondrosophilid insects examined so far, express a second cry gene (designated insect CRY2) that encodes a vertebrate-like CRY that is also expressed in pars lateralis. An ancestral circadian clock mechanism has been defined in monarchs, in which CRY1 functions as a blue light photoreceptor for photic entrainment, whereas CRY2 functionswithin the clockwork as themajor transcriptional repressor of an intracellular negative transcriptional feedback loop. A CRY1-staining neural pathway has been identified that may connect the circadian (navigational) clock to polarized light input important for sun compass navigation, and a CRY2-positive neural pathway has been discovered that may communicate circadian information directly from the circadian clock to the central complex, the likely site of the sun compass. The monarch butterfly may thus use the CRY proteins as components of the circadian mechanism and also as output molecules that connect the clock to various aspects of the sun compass apparatus. PMID- 18419269 TI - Structure and function of animal cryptochromes. AB - Cryptochrome (CRY) is a photolyase-like flavoprotein with no DNA-repair activity but with known or presumed blue-light receptor function. Animal CRYs have DNA binding and autokinase activities, and their flavin cofactor is reduced by photoinduced electron transfer. In Drosophila, CRY is a major circadian photoreceptor, and in mammals, the two CRY proteins are core components of the molecular clock and potential circadian photoreceptors. In mammals, CRYs participate in cell cycle regulation and the cellular response to DNA damage by controlling the expression of some cell cycle genes and by directly interacting with checkpoint proteins. PMID- 18419270 TI - Structure function analysis of mammalian cryptochromes. AB - Members of the photolyase/cryptochrome family are flavoproteins that share an extraordinary conserved core structure (photolyase homology region, PHR), but the presence of a carboxy-terminal extension is limited to the cryptochromes. Photolyases are DNA-repair enzymes that remove UV-light-induced lesions. Cryptochromes of plants and Drosophila act as circadian photoreceptors, involved in light entrainment of the biological clock. Using knockout mouse models, mammalian cryptochromes (mCRY1 and mCRY2) were identified as essential components of the clock machinery. Within the mammalian transcription-translation feedback loop generating rhythmic gene expression, mCRYs potently inhibit the transcription activity of the CLOCK/BMAL1 heterodimer and protect mPER2 from 26S protesome-mediated degradation. By analyzing a set of mutant mCRY1 proteins and photolyase/mCRY1 chimeric proteins, we found that the carboxyl terminus has a determinant role in mCRY1 function by harboring distinguished domains involved in nuclear import and interactions with other clock proteins. Moreover, the carboxyl terminus must cross-talk with the PHR to establish full transcription repression capacity in mCRY1. We propose that the presence of the carboxyl terminus in cryptochromes, which varies in sequence composition among mammalian, Drosophila, and plant CRYs, is critical for their different functions and possibly contributed to shape the different architecture and biochemistry of the clock machineries in these organisms. PMID- 18419271 TI - The Gonyaulax clock at 50: translational control of circadian expression. AB - The unicellular circadian clock of Gonyaulax polyedra (now renamed Lingulodinium polyedrum) has provided important insights concerning circadian rhythmicity. Many, perhaps most, of its key systems are circadian-controlled, ranging from bioluminescence and photosynthesis to motility, cell division, and the synthesis of many proteins, favoring the "master clock" concept. But different rhythms may have different free-running periods and different phase angles under different T cycles, observations not easily accommodated in a single oscillator model. Gonyaulax has a feature significantly different from that of other known systems, namely, that clock control of protein synthesis occurs at the translational level. With one mRNA, this involves a protein binding to a 22-nucleotide region in the 3'-untranslated region (3'UTR), but no similar regions have been found in other mRNAs. Pulses of protein synthesis inhibitors cause phase shifts, whereas inhibitors of protein phosphorylation administered chronically cause period changes. In Gonyaulax and other systems, low temperature results in arrhythmicity. A return to a permissive temperature results in a reinitiation of the rhythm, with the phase established by the time of increase, similar to the effect of bright light. Evidence for cellular communication via substance(s) in the medium has been obtained. PMID- 18419272 TI - Posttranscriptional regulation of mammalian circadian clock output. AB - Circadian clocks are present in many different cell types/tissues and control many aspects of physiology. This broad control is exerted, at least in part, by the circadian regulation of many genes, resulting in rhythmic expression patterns of 5-10% of the mRNAs in a given tissue. Although transcriptional regulation is certainly involved in this process, it is becoming clear that posttranscriptional mechanisms also have important roles in producing the appropriate rhythmic expression profiles. In this chapter, we review the available data about posttranscriptional regulation of circadian gene expression and highlight the potential role of Nocturnin (Noc) in such processes. NOC is a deadenylase-a ribonuclease that specifically removes poly(A) tails from mRNAs-that is expressed widely in the mouse with high-amplitude rhythmicity. Deadenylation affects the stability and translational properties of mRNAs. Mice lacking the Noc gene have metabolic defects including a resistance to diet-induced obesity, decreased fat storage, changes in lipid-related gene expression profiles in the liver, and altered glucose and insulin sensitivities. These findings suggest that NOC has a pivotal role downstream from the circadian clockwork in the post-transcriptional regulation genes involved in the circadian control of metabolism. PMID- 18419273 TI - Biological clocks and the coordination theory of RNA operons and regulons. AB - One of the regulatory models of circadian rhythms involves the oscillation of transcription and translation. Although transcription factors have been widely examined during circadian processes, posttranscriptional mechanisms are less well studied. Several laboratories have used microarrays to detect changes in mRNA expression throughout the circadian cycle and have found that mRNAs encoding the RNA-binding proteins (RBPs) nocturnin and butyrate response factor (BRF1) undergo rhythmic changes. Nocturnin is a deadenylation enzyme that removes poly(A) from the 3' ends of mRNAs, whereas BRF1 destabilizes mRNAs encoding early response gene (ERG) transcripts that contain AU-rich sequences in their 3'-untranslated regions (UTRs). Moroni and coworkers proposed that BRF1 functions as an oscillating posttranscriptional RNA operon (PTRO) that diurnally degrades ERG transcripts in peripheral organs (Keene and Tenenbaum 2002; Benjamin et al. 2006). The PTRO model posits that mRNAs can be members of one or more discrete functionally related subsets of mRNAs as determined by cis elements in mRNA and trans-acting RBPs or microRNAs that collectively recognize these cis elements (Keene 2007). This chapter describes the basis of posttranscriptional coordination by RNA operons and their potential for horizontal transfer among cells and discusses the potential for RBPs and microRNAs to participate in coordinating circadian rhythms and other biological clocks. PMID- 18419274 TI - Role of phosphorylation in the mammalian circadian clock. AB - Circadian clocks regulate a wide variety of processes ranging from gene expression to behavior. At the molecular level, circadian rhythms are thought to be produced by a set of clock genes and proteins interconnected to form transcriptional-translational feedback loops. Rhythmic gene expression was formerly regarded as the major drive for rhythms in clock protein abundance, but recent findings underline the crucial importance of posttranslational mechanisms for both the generation and dynamics of circadian rhythms. In particular, the reversible phosphorylation of PER proteins-essential components within the negative feedback loop in Drosophila and mammals-seems to have a key role for the correct timing of nuclear repression. To understand how PER protein phosphorylation regulates the dynamics of the circadian oscillator, we have mapped endogenous phosphorylation sites in mPER2. Detailed investigation of the functional role of one particular phosphorylation site (Ser-659, which is mutated in the familial advanced sleep phase syndrome [FASPS]) led us propose a model of functionally different phosphorylation sites in PER2. This concept explains not only the FASPS phenotype, but also the effect of the tau mutation in hamster. PMID- 18419276 TI - Posttranslational control of the Neurospora circadian clock. AB - The eukaryotic circadian clocks are composed of autoregulatory circadian negative feedback loops that include both positive and negative elements. Investigations of the Neurospora circadian clock system have elucidated many of the basic mechanisms that underlie circadian rhythms, including negative feedback and light and temperature entrainment common to all eukaryotic clocks. The conservation of the posttranslational regulators in divergent circadian systems suggests that the processes mediating the modification and degradation of clock proteins may be the common foundation that allows the evolution of circadian clocks in eukaryotic systems. In this chapter, we summarize recent studies of the Neurospora circadian clock with emphasis on posttranslational regulation in the circadian negative feedback loop. PMID- 18419275 TI - Posttranslational regulation of Neurospora circadian clock by CK1a-dependent phosphorylation. AB - Frequency (FRQ) and its transcriptional activator, the White Collar Complex (WCC), are essential components of interconnected feedback loops of the circadian clock of Neurospora. In a negative feedback loop, FRQ inhibits the WCC by recruiting casein kinase 1a (CK1a) and supporting its phosphorylation. In an interconnected positive loop, FRQ supports accumulation of high levels of WCC. Phosphorylation of clock proteins is crucial for the temporal and spatial coordination of these functions. We identified three isoforms of CK1a generated by alternative splicing that all interact with FRQ. Furthermore, we show that WC 2 is phosphorylated by CK1a in vitro and that WC-2 phosphorylation is inhibited in vivo by the CK1-specific inhibitor IC261. Finally, we demonstrate that CK1a activity regulates levels of WC-2. PMID- 18419277 TI - Posttranslational photomodulation of circadian amplitude. AB - The transcription-translation feedback loops that form our current view of how the core mechanism of the clock operates is being challenged, as more and more posttranslational events are seen as essential to a full understanding of oscillator function. But in addition to phosphorylation, other processes may be involved. Here, a novel mechanism of posttranslational photomodulation of circadian amplitude is described that uniquely ties together light perception, protein stabilization, and proteolysis. In the process, the waveform of a core clock component is sharpened or "sculpted," resulting in appropriately high amplitude and proper phasing to obtain normal clock function. PMID- 18419279 TI - Principles and problems revolving around rhythm-related genetic variants. AB - Much of what is known about the regulation of circadian rhythms has stemmed from the induction, recognition, or manufacture of genetic variants. Such investigations have been especially salient in chronobiological analyses of Drosophila. Many starting points for elucidation of rhythmic processes operating in this insect entailed the isolation of mutants or the design of engineered gene modifications. Various features of the principles and practices associated with the genetic approach toward understanding clock functions, and chronobiologically related ones, are discussed from perspectives that are largely genetic as such, although intertwined with certain neurogenetic and molecular-genetic concerns when appropriate. Key themes in this treatment connect with the power and problems associated with multiply mutant forms of rhythm-related genes, with the opportunistic or problematical aspects of multigenic variants that are in play (sometimes surprisingly), and with a question as to how forceful chronogenetic inferences have been in terms of elucidating the mechanisms of circadian pacemaking. PMID- 18419278 TI - Circadian output, input, and intracellular oscillators: insights into the circadian systems of single cells. AB - Circadian output comprises the business end of circadian systems in terms of adaptive significance. Work on Neurospora pioneered the molecular analysis of circadian output mechanisms, and insights from this model system continue to illuminate the pathways through which clocks control metabolism and overt rhythms. In Neurospora, virtually every strain examined in the context of rhythms bears the band allele that helps to clarify the overt rhythm in asexual development. Recent cloning of band showed it to be an allele of ras-1 and to affect a wide variety of signaling pathways yielding enhanced light responses and asexual development. These can be largely phenocopied by treatments that increase levels of intracellular reactive oxygen species. Although output is often unidirectional, analysis of the prd-4 gene provided an alternative paradigm in which output feeds back to affect input. prd-4 is an allele of checkpoint kinase 2 that bypasses the requirement for DNA damage to activate this kinase; FRQ is normally a substrate of activated Chk2, so in Chk2(PRD-4), FRQ is precociously phosphorylated and the clock cycles more quickly. Finally, recent adaptation of luciferase to fully function in Neurospora now allows the core FRQ/WCC feedback loop to be followed in real time under conditions where it no longer controls the overt rhythm in development. This ability can be used to describe the hierarchical relationships among FRQ-Less Oscillators (FLOs) and to see which are connected to the circadian system. The nitrate reductase oscillator appears to be connected, but the oscillator controlling the long-period rhythm elicited upon choline starvation appears completely disconnected from the circadian system; it can be seen to run with a very long noncompensated 60-120-hour period length under conditions where the circadian FRQ/WCC oscillator continues to cycle with a fully compensated circadian 22-hour period. PMID- 18419280 TI - Synchronization of the Drosophila circadian clock by temperature cycles. AB - The natural light/dark and temperature cycles are considered to be the most prominent factors that synchronize circadian clocks with the environment. Understanding the principles of temperature entrainment significantly lags behind our current knowledge of light entrainment in any organism subject to circadian research. Nevertheless, several effects of temperature on circadian clocks are well understood, and similarities as well as differences to the light-entrainment pathways start to emerge. This chapter provides an overview of the temperature effects on the Drosophila circadian clock with special emphasis on synchronization by temperature cycles. As in other organisms, such temperature cycles can serve as powerful time cues to synchronize the clock. Mutants that specifically interfere with aspects of temperature entrainment have been isolated and will likely help to reveal the underlying mechanisms. These mechanisms involve transcriptional and posttranscriptional regulation of clock genes. For synchronization of fly behavior by temperature cycles, the generation of a whole organism or systemic signal seems to be required, even though individual fly tissues can be synchronized under isolated culture conditions. If true, the requirement for such a signal would reveal a fundamental difference to the light entrainment mechanism. PMID- 18419283 TI - Novel insights from genetic and molecular characterization of the human clock. AB - Biological rhythms govern the ebb and flow of life on planet Earth. Animals have an internal timekeeping mechanism that precisely regulates 24-hour rhythms of body function and behavior and synchronizes them to the day/night cycle. Circadian pacemakers trigger behavioral and physiological processes that dictate our daily rhythms. Despite the importance of the circadian clock to all aspects of our physiology and behavior, the opportunity to probe the human circadian clock only recently became possible with the recognition of Mendelian circadian variants in people (familial advanced sleep phase syndrome, FASPS). We have now cloned several genes and identified mutations causing FASPS. Study of these genes and the proteins they encode and engineering of the human mutations into mouse models are allowing study of this fascinating phenotype and yielding novel insights into circadian regulation in humans. Ultimately, such work will allow us to understand the similarities and differences between the human clock and those of model organisms. In addition, recent studies have also linked disruption of the circadian clock with numerous ailments, including cancer, cardiovascular diseases, asthma, and learning disorders. Thus, studying the molecular mechanism of human circadian rhythmicity will have an enormous impact on our understanding of human health and disease. It should also lead to new strategies for pharmacological manipulation of the human clock to improve the treatment of jet lag, various clock-related sleep and psychiatric disorders, and other human diseases. PMID- 18419281 TI - What is there left to learn about the Drosophila clock? AB - Circadian rhythms offer probably the best understanding of how genes control behavior, and much of this understanding has come from studies in Drosophila. More recently, genetic manipulation of clock neurons in Drosophila has helped identify how daily patterns of activity are programmed by different clock neuron groups. Here, we review some of the more recent findings on the fly molecular clock and ask what more the fly model can offer to circadian biologists. PMID- 18419282 TI - Genetics and neurobiology of circadian clocks in mammals. AB - In animals, circadian behavior can be analyzed as an integrated system, beginning with genes and leading ultimately to behavioral outputs. In the last decade, the molecular mechanism of circadian clocks has been unraveled primarily by the use of phenotype-driven (forward) genetic analysis in a number of model systems. Circadian oscillations are generated by a set of genes forming a transcriptional autoregulatory feedback loop. In mammals, there is a "core" set of circadian genes that form the primary negative feedback loop of the clock mechanism (Clock/Npas2, Bmal1, Per1, Per2, Cry1, Cry2, and CK1epsilon). A further dozen candidate genes have been identified and have additional roles in the circadian gene network such as the feedback loop involving Rev-erbalpha. Despite this remarkable progress, it is clear that a significant number of genes that strongly influence and regulate circadian rhythms in mammals remain to be discovered and identified. As part of a large-scale N-ethyl-N-nitrosourea mutagenesis screen using a wide range of nervous system and behavioral phenotypes, we have identified a number of new circadian mutants in mice. Here, we describe a new short-period circadian mutant, part-time (prtm), which is caused by a loss-of function mutation in the Cryptochrome1 (Cry1) gene. We also describe a long period circadian mutant named Overtime (Ovtm). Positional cloning and genetic complementation reveal that Ovtm is encoded by the F-box protein FBXL3, a component of the SKP1-CUL1-F-box protein (SCF) E3 ubiquitin ligase complex. The Ovtm mutation causes an isoleucine to threonine (I364T) substitution leading to a loss of function in FBXL3 that interacts specifically with the CRYPTOCHROME (CRY) proteins. In Ovtm mice, expression of the PERIOD proteins PER1 and PER2 is reduced; however, the CRY proteins CRY1 and CRY2 are unchanged. The loss of FBXL3 function leads to a stabilization of the CRY proteins, which in turn leads to a global transcriptional repression of the Per and Cry genes. Thus, Fbxl3(Ovtm) defines a molecular link between CRY turnover and CLOCK/BMAL1-dependent circadian transcription to modulate circadian period. PMID- 18419284 TI - Circadian entrainment of Neurospora crassa. AB - The circadian clock evolved under entraining conditions, yet most circadian experiments and much circadian theory are built around free-running rhythms. The interpretation of entrainment experiments is certainly more complex than that of free-running rhythms due to the relationship between exogenous and endogenous cycles. Here, we systematically describe entrainment in the simplest of the traditional eukaryotic model systems in circadian research, Neurospora crassa. This fungus forms a mass of spores (bands of conidia) each day. Over a wide range of photoperiods, these bands begin to appear at midnight, suggesting integration of neither dawn nor dusk signals alone. However, when symmetrical light/dark cycles (T cycles, each with 50% light) are applied, dusk determines the time of conidiation with a uniform, period-dependent delay in phase. This "forced" synchronization appears to be specific for the zeitgeber light because similar experiments, but using temperature, result in systematic entrainment, with bands appearing relatively later in shorter cycles and earlier in longer cycles. We find that the molecular mechanism of entrainment primarily concerns posttranscriptional regulation. Finally, we have used Neurospora to investigate acute effects of zeitgeber stimuli known as "masking." PMID- 18419285 TI - Constant darkness is a mammalian biological signal. AB - Environmental light is a potent modulator of mammalian circadian rhythm and expression of clock genes. Constant darkness (DD) is regarded as a "free-running" circadian state. In nature, hibernating mammals encounter constant darkness (DD) seasonally. Circadian expression of enzymes involved in fat catabolism, procolipase (CLP) and pancreatic-lipase-related protein 2 (PLRP2), were identified in many peripheral organs of mice during DD but not during regular light/dark (LD) cycles. Circulating 5'-adenosine monophosphate (5'-AMP) was associated with DD-activated gene expression. Synthetic 5'-AMP, when injected into LD mice, activated procolipase expression in their peripheral organs and the animals become severely hypothermic, both key features of hibernating mammals. These findings identified a circadian-regulated metabolic cycle in mammals that may be associated with hypometabolic behaviors such as hibernation and torpor. PMID- 18419286 TI - Entrainment of the human circadian clock. AB - Humans are an excellent model system for studying entrainment of the circadian clock in the real world. Unlike the situation in laboratory experiments, entrainment under natural conditions is achieved by different external signals as well as by internal signals generated by multiple feedbacks within the system (e.g., behavior-dependent light and temperature changes, melatonin levels, or regular nutrient intake). Signals that by themselves would not be sufficient zeitgebers may contribute to entrainment in conjunction with other self sufficient zeitgeber signals (e.g., light). The investigation of these complex zeitgeber interactions seems to be problematic in most model systems and strengthens the human system for circadian research. Here, we review our endeavors measuring human entrainment in real life, predominantly with the help of the Munich ChronoType Questionnaire (MCTQ). The large number of participants in our current MCTQ database allows accurate quantification of the human phase of entrainment (chronotype) and how it depends on age or sex. We also present new data showing how chronotype depends on natural light exposure. The results indicate the importance of zeitgeber strength on human entrainment and help in understanding the differences in chronotype, e.g., between urban and rural regions. PMID- 18419287 TI - Peripheral clocks: keeping up with the master clock. AB - Circadian clocks influence most aspects of physiology and behavior, so perhaps it is not surprising that circadian oscillators exist in nearly all mammalian cells. These cells remain synchronized to the outside world in hierarchical fashion, with a "master clock" tissue in the suprachiasmatic nucleus of the hypothalamus receiving light input from the retina and then conveying timing information to "slave" clocks in peripheral tissues. Recent research has highlighted both the similarities and differences between central and peripheral clocks and provided new insight into their communication. Above all, however, this parallelism of clockwork has provided a unique opportunity to study at the cellular level a regulatory mechanism that affects complex behaviors. PMID- 18419288 TI - Physiological importance of a circadian clock outside the suprachiasmatic nucleus. AB - Circadian clocks are widely distributed in mammalian tissues, but little is known about the physiological functions of clocks outside the suprachiasmatic nucleus of the brain. The retina has an intrinsic circadian clock, but its importance for vision is unknown. Here, we show that mice lacking Bmal1, a gene required for clock function, had abnormal retinal transcriptional responses to light and defective inner retinal electrical responses to light, but normal photoreceptor responses to light and retinas that appeared structurally normal as observed by light and electron microscopy. We generated mice with a retina-specific genetic deletion of Bmal1, and they had defects of retinal visual physiology essentially identical to those of mice lacking Bmal1 in all tissues and lacked a circadian rhythm of inner retinal electrical responses to light. Our findings indicate that the intrinsic circadian clock of the retina regulates retinal visual processing in vivo. PMID- 18419289 TI - Regulation of circadian gene expression in liver by systemic signals and hepatocyte oscillators. AB - The mammalian circadian timing system has a hierarchical structure, in that a master pacemaker located in the suprachiasmatic nuclei (SCN) coordinates slave oscillators present in virtually all body cells. In both the SCN and peripheral organs, the rhythm-generating oscillators are self-sustained and cell-autonomous, and it is likely that the molecular makeup of master and slave oscillators is nearly identical. However, due to variations in period length, the phase coherence between peripheral oscillators in intact animals must be established by daily signals emanating directly or indirectly from the SCN master clock. The synchronization of individual cellular clocks in peripheral organs is probably accomplished by immediate-early genes that interpret the cyclic systemic signals and convey this phase information to core clock components. This model predicts that circadian gene expression in peripheral organs can be influenced either by systemic signals emanating from the SCN master clock, local oscillators, or both. We developed a transgenic mouse strain in which hepatocyte clocks are only operative when the tetracycline analog doxycycline is added to the food or drinking water. The genome-wide mapping of genes whose cyclic expression in liver does not depend on functional hepatocyte oscillators unveiled putative signaling pathways that may participate in the phase entrainment of peripheral clocks. PMID- 18419291 TI - The yeast metabolic cycle: insights into the life of a eukaryotic cell. AB - The budding yeast Saccharomyces cerevisiae undergoes robust oscillations in oxygen consumption during continuous growth under nutrient-limited conditions. Comprehensive microarray studies reveal that more than half of the yeast genome is expressed periodically as a function of these respiratory oscillations, thereby specifying an extensively orchestrated program responsible for regulating numerous cellular outputs. Here, we summarize the logic of the yeast metabolic cycle (YMC) and highlight additional cellular processes that are predicted to be compartmentalized in time. Certain principles of temporal orchestration as seen during the YMC might be conserved across other biological cycles. PMID- 18419290 TI - Integrating the circadian oscillator into the life of the cyanobacterial cell. AB - In two decades, the study of circadian rhythms in cyanobacteria has gone from observations of phenomena in intractable species to the development of a model organism for mechanistic study, atomic-resolution structures of components, and reconstitution of a circadian biochemical oscillation in vitro. With sophisticated biochemical, biophysical, genetic, and genomic tools in place, the circadian clock of the unicellular cyanobacterium Synechococcus elongatus is poised to be the first for which a systems-level understanding can be achieved. PMID- 18419292 TI - Complexity of the Neurospora crassa circadian clock system: multiple loops and oscillators. AB - Organisms from bacteria to humans use a circadian clock to control daily biochemical, physiological, and behavioral rhythms. We review evidence from Neurospora crassa that suggests that the circadian clock is organized as a network of genes and proteins that form coupled evening- and morning-specific oscillatory loops that can function autonomously, respond differently to environmental inputs, and regulate phase-specific outputs. There is also evidence for coupled morning and evening oscillator loops in plants, insects, and mammals, suggesting conservation of clock organization. From a systems perspective, fungi provide a powerful model organism for investigating oscillator complexity, communication between oscillators, and addressing reasons why the system has evolved to be so complex. PMID- 18419293 TI - The DIURNAL project: DIURNAL and circadian expression profiling, model-based pattern matching, and promoter analysis. AB - The DIURNAL project ( http://diurnal.cgrb.oregonstate.edu/ ) provides a graphical interface for mining and viewing diurnal and circadian microarray data for Arabidopsis thaliana, poplar, and rice. The database is searchable and provides access to several user-friendly Web-based data-mining tools with easy-to understand output. The associated tools include HAYSTACK ( http://haystack.cgrb.oregonstate.edu/ ) and ELEMENT ( http://element.cgrb.oregonstate.edu/ ). HAYSTACK is a model-based pattern matching algorithm for identifying genes that are coexpressed and potentially coregulated. HAYSTACK can be used to analyze virtually any large-scale microarray data set and provides an alternative method for clustering microarray data from any experimental system by grouping together genes whose expression patterns match the same or similar user-defined patterns. ELEMENT is a Web-based program for identifying potential cis-regulatory elements in the promoters of coregulated genes in Arabidopsis, poplar, and rice. Together, DIURNAL, HAYSTACK, and ELEMENT can be used to facilitate cross-species comparisons among the plant species supported and to accelerate functional genomics efforts in the laboratory. PMID- 18419294 TI - Systems biology of mammalian circadian clocks. AB - Systems Biology is a natural extension of molecular biology and can be defined as biology after identification of key gene(s). Systems-biological research is hence seen as a multistage process, beginning with the comprehensive identification and quantitative analysis of individual system components and their networked interactions and leading to the ability to control existing systems toward the desired state and design new ones based on an understanding of structure and underlying dynamical principles. In this chapter, we take mammalian circadian clocks as a model system and describe systems-biological approaches, including the identification of clock-controlled genes, clock-controlled cis elements, and clock transcriptional circuits driven by functional genomics; the parameter change of clock components followed by quantitative measurement; and the dynamic and quantitative perturbation of the clock and its application to one of the fundamental but yet-unsolved questions: singularity behavior of clocks. As perspective for systems-biological investigations, we also introduce the system level dynamical questions related to the core of clocks, including delay, nonlinearity, temperature-compensation and synchronization of mammalian circadian oscillator(s), and the system-level information problems related to clocks in the environment, including the internal representation of light change through perfect adaptation and internal representation of day length through photoperiodism in mammals. PMID- 18419295 TI - High-resolution time course analysis of gene expression from pituitary. AB - In both the suprachiasmatic nucleus (SCN) and peripheral tissues, the circadian oscillator drives rhythmic transcription of downstream target genes. Recently, a number of studies have used DNA microarrays to systematically identify oscillating transcripts in plants, fruit flies, rats, and mice. These studies have identified several dozen to many hundred rhythmically expressed genes by sampling tissues every 4 hours for 1, 2, or more days. To extend this work, we have performed DNA microarray analysis on RNA derived from the mouse pituitary sampled every hour for 2 days. COSOPT and Fisher's G-test were used at a false discovery rate of less than 5% to identify more than 250 genes in the pituitary that oscillate with a 24-hour period length. We found that increasing the frequency of sampling across the circadian day dramatically increased the statistical power of both COSOPT and Fisher's G-test, resulting in considerably more high-confidence identifications of rhythmic transcripts than previously described. Finally, to extend the utility of these data sets, a Web-based resource has been constructed (at http://wasabi.itmat.upenn.edu/circa/mouse ) that is freely available to the research community. PMID- 18419296 TI - Nuclear receptors, metabolism, and the circadian clock. AB - As ligand-dependent transcription factors, the nuclear receptor superfamily governs a remarkable array of rhythmic physiologic processes such as metabolism and reproduction. To provide a "molecular blueprint" for nuclear receptor function in circadian biology, we established a diurnal expression profile of all mouse nuclear receptors in critical metabolic tissues. Our finding of broad expression and tissue-specific oscillation of nuclear receptors along with their key target genes suggests that diurnal nuclear receptor expression may contribute to established rhythms in metabolic physiology and that nuclear receptors may be involved in coupling peripheral circadian clocks to divergent metabolic outputs. Conversely, nuclear receptors may serve peripheral clock input pathways, integrating signals from the light-sensing central clock in the suprachiasmatic nucleus and other environmental cues, such as nutrients and xenobiotics. Interplay between the core circadian clock and nuclear receptors may define a large-scale signaling network that links biological timing to metabolic physiology. PMID- 18419297 TI - Bacterial circadian programs. AB - Twenty years ago, it was widely believed that prokaryotes were too "simple" to have evolved circadian programs. Since that time, however, the cyanobacterial circadian system has progressed from a curiosity to a major model system for analyzing clock phenomena. In addition to globally regulating gene expression, cyanobacteria are one of the only systems in which the adaptive fitness of a circadian system has been rigorously evaluated. Moreover, cyanobacteria are the only clock system in which all essential proteins of the core oscillator have been crystallized and structurally determined, namely, the KaiA, KaiB, and KaiC proteins. A biochemical oscillator can be reconstituted in vitro with these three purified Kai proteins and displays the key properties of temperature-compensated rhythmicity. This result spectacularly demonstrates that a strictly posttranslational clock is sufficient to elaborate circadian phenomena and that a transcription-translation feedback loop is not obligatory. The conjunction of structural information on essential clock proteins with a defined system that reconstitutes circadian oscillations in vitro leads to a turning point whereby biophysical and biochemical approaches bring analyses of circadian clock-work to an unprecedented level of molecular detail. PMID- 18419298 TI - Stochastic phase oscillators and circadian bioluminescence recordings. AB - Cultured circadian oscillators from peripheral tissues were recently shown to be both cell-autonomous and self-sustained. Therefore, the dominant cause for amplitude reduction observed in bioluminescence recordings of cultured fibroblasts is desynchronization, rather than the damping of individual oscillators. Here, we review a generic model for quantifying luminescence signals from biochemical oscillators, based on noisy-phase oscillators. Our model incorporates three essential features of circadian clocks: the stability of the limit cycle, fluctuations, and intercellular coupling. The model is then used to analyze bioluminescence recordings from immortalized and primary fibroblasts. Fits to population recordings allow simultaneous estimation of the stability of the limit cycle (or equivalently, the stiffness of individual frequencies), the period dispersion, and the interaction strength between cells. Consistent with other work, coupling is found to be weak and insufficient to synchronize cells. Interestingly, we find that frequency fluctuations remain correlated for longer periods than one clock cycle, which is confirmed from individual cell recordings. We discuss briefly how to link the generic model with more microscopic models, which suggests mechanisms by which circadian oscillators resist fluctuations and maintain accurate timing in the periphery. PMID- 18419299 TI - Reversible protein phosphorylation regulates circadian rhythms. AB - Protein phosphorylation regulates the period of the circadian clock within mammalian cells. Circadian rhythms are an approximately 24-hour cycle that regulates key biological processes. Daily fluctuations of wakefulness, stress hormones, lipid metabolism, immune function, and the cell division cycle are controlled by the molecular clocks that function throughout our bodies. Mutations in regulatory components of the clock can shorten or lengthen the timing of the rhythms and have significant physiological consequences. The clock is formed by a negative feedback loop of transcription, translation, and inhibition of transcription. The precision of clock timing is controlled by protein kinases and phosphatases. Casein kinase Iepsilon is a protein kinase that regulates the circadian clock by periodic phosphorylation of the proteins PER1 and PER2, controlling their stability and localization. The role of phosphorylation in regulating PER function in the clock has been explored in detail. Quantitative modeling has proven to be very useful in making important predictions about how changes in phosphorylation alter the clock's behavior. Quantitative data from biological studies can be used to refine the quantitative model and make additional testable predictions. A detailed understanding of how reversible protein phosphorylation regulates circadian rhythms and a detailed quantitative model that makes clear, testable, and accurate predictions about the clock and how we may manipulate it can have important benefits for human health. Pharmacological manipulation of rhythms could mitigate stress from jet lag, shift work, and perhaps even seasonal affective disorder. PMID- 18419300 TI - Evolution of the clock from yeast to man by period-doubling folds in the cellular oscillator. AB - Analysis of genome-wide oscillations in transcription reveals that the cell is an oscillator and an attractor and that the maintenance of a stable phenotype requires that maximums in expression in clusters of transcripts must be poised at antipodal phases around the steady state-this is the dynamic architecture of phenotype. Plots of the path through concentration phase space taken by all of the transcripts of Saccharomyces cerevisiae yield a simple three-dimensional surface. How this surface might change as period lengthens or as a cell differentiates is at the center of current work. We have shown that changes in gene expression in response to mutation or perturbation by drugs occur through a folding or unfolding of the surface described by this circle of transcripts and we suggest that the path from this 40-minute oscillation to the cell cycle and circadian rhythms takes place through a series of period-two or period-three bifurcations. These foldings in the surface of the putative attractor result in an increasingly dense set of nested trajectories in the concentrations of message and protein. Evolutionary advantage might accrue to an organism that could change period by changes in just one or a few genes as day length increased from 4 hours in the prebiotic Earth, through 8 hours during the expansion of photoautotrophs, to the present 24 hours. PMID- 18419301 TI - Intracellular developmental timers. AB - One of the most poorly understood aspects of animal development is how the timing of developmental events is controlled. In most vertebrate cell lineages, for example, precursor cells divide a limited number of times before they stop and terminally differentiate, but it is not known what controls when the cells stop dividing and differentiate. There is increasing evidence, however, that intracellular timers play an important part. Such cell-intrinsic timers are examples of intracellular developmental programs that change precursor cells over time. My colleagues and I have studied such intracellular timers and programs in rodent oligodendrocyte precursor cells (OPCs), as reviewed here. PMID- 18419303 TI - Building the spine: the vertebrate segmentation clock. AB - One of the most striking characteristics of many animal and plant species is their organization in a series of periodically repeated anatomical modules. In animals, this particular patterning strategy of the body axis is termed segmentation, and it is observed in both vertebrates and invertebrates. Vertebrate segmentation has been associated with a molecular oscillator-the segmentation clock-whose existence had been predicted on theoretical grounds in the clock and wave-front model. The segmentation clock is proposed to generate pulses of signaling used for the positioning of segmental boundaries. Whereas several models have proposed that simple negative autoregulatory circuits involving the transcription repressors of the hairy and enhancer of split family constitute the clock pacemaker, recent microarray studies in mouse have identified a large network of oscillating signaling genes belonging to the Notch, Wnt, and FGF (fibroblast growth factor) pathways. Thus, significant progress has been made, but the molecular nature of the clockwork underlying the oscillator remains poorly understood. Few examples of oscillators exist in developmental biology, and the segmentation clock provides a unique model of periodic regulation in patterning. PMID- 18419302 TI - Transcriptional feedback loop regulation, function, and ontogeny in Drosophila. AB - The Drosophila circadian oscillator is composed of interlocked period/timeless (per/tim) and Clock (Clk) transcriptional feedback loops. These feedback loops drive rhythmic transcription having peaks at dawn and dusk during the daily cycle and function in the brain and a variety of peripheral tissues. To understand how the circadian oscillator keeps time and controls metabolic, physiological, and behavioral rhythms, we must determine how these feedback loops regulate rhythmic transcription, determine the relative importance of the per/tim and Clk feedback loops with regard to circadian oscillator function, and determine how these feedback loops come to be expressed in only certain tissues. Substantial insight into each of these issues has been gained from experiments performed in our lab and others and is summarized here. PMID- 18419304 TI - Ultradian oscillators in somite segmentation and other biological events. AB - Somite formation occurs every 2 hours in mouse embryos by periodic segmentation of the anterior ends of the presomitic mesoderm, and this process is controlled by a biological clock called the segmentation clock. During this process, the basic helix-loop-helix gene Hes7 is cyclically expressed, and each cycle leads to generation of a bilateral pair of somites. Both sustained expression and loss of expression of Hes7 result in severe somite fusion, indicating that Hes7 constitutes an essential component of the segmentation clock. Interestingly, expression of the related gene Hes1 also oscillates with a periodicity of about 2 hours in many cell types. Both sustained Hes1 expression and loss of Hes1 activity lead to retardation of the G(1) phase of the cell cycle, suggesting that Hes1 oscillation with an ultradian rhythm is required for efficient cell proliferation. Both Hes1 and Hes7 oscillations are regulated by negative feedback and rapid degradation of their gene products. Strikingly, expression of other factors such as Stat-Socs and Smad signaling molecules also display ultradian rhythms. All of these data suggest that ultradian oscillations are more general responses than were previously thought and that oscillatory and sustained gene expression results in different biological outcomes. PMID- 18419305 TI - The role of circadian regulation in cancer. AB - Proper circadian regulation is essential for the well being of the organism, and disruption of circadian rhythms is associated with pathological conditions including cancer. In mammals, the core clock genes, Per1 and Per2, are key regulators of circadian rhythms both in the central clock in the hypothalamous and in peripheral tissues. Recent findings revealed molecular links between Per genes and cellular components that control fundamental cellular processes such as cell division and DNA damage. New data also shed light on mechanisms by which circadian oscillators operate in peripheral organs to influence tissue-dependent metabolic and hormonal pathways. Circadian cycles are linked to basic cellular functions, as well as to tissue-specific processes through the control of gene expression and protein interactions. By controlling global networks such as chromatin remolding and protein families, which themselves regulate a broad range of cellular functions, circadian regulation impinges upon almost all major physiological pathways. These molecular insights illustrate how disregulation of circadian rhythms might influence the susceptibility to cancer development and provide further support for the emerging role of circadian genes in tumor suppression. PMID- 18419306 TI - Cross-talks between circadian timing system and cell division cycle determine cancer biology and therapeutics. AB - The circadian clock orchestrates cellular functions over 24 hours, including cell divisions, a process that results from the cell cycle. The circadian clock and cell cycle interact at the level of genes, proteins, and biochemical signals. The disruption or the reinforcement of the host circadian timing system, respectively, accelerates or slows down cancer growth through modifications of host and tumor circadian clocks. Thus, cancer cells not only display mutations of cell cycle genes but also exhibit severe defects in clock gene expression levels or 24-hour patterns, which can in turn favor abnormal proliferation. Most of the experimental research actively ongoing in this field has been driven by the original demonstration that cancer patients with poor circadian rhythms had poor quality of life and poor survival outcome independently of known prognostic factors. Further basic research on the gender dependencies in circadian properties is now warranted, because a large clinical trial has revealed that gender can largely affect the survival outcome of cancer patients on chronotherapeutic delivery. Mathematical models further show that the therapeutic index of chemotherapeutic drugs can be optimized through distinct delivery profiles, depending on the initial host/tumor status and variability in circadian entrainment and/or cell cycle length. Clinical trials and systems-biology approaches in cancer chronotherapeutics raise novel issues to be addressed experimentally in the field of biological clocks. The challenge ahead is to therapeutically harness the circadian timing system to concurrently improve quality of life and down-regulate malignant growth. PMID- 18419307 TI - The clock proteins, aging, and tumorigenesis. AB - Many aspects of mammalian physiology and behavior are driven by an intrinsic timekeeping system that has an important role in synchronizing various biological processes within an organism and coordinating them with the environment. It is believed that deregulation of this coordination may cause the development of various pathologies. However, recent studies using mice deficient in individual components of the circadian system clearly demonstrated more complex interaction of the circadian system with various biological processes. The growing amount of evidence suggests that in addition to their roles in the core clock mechanism, some of the components of the molecular oscillator are involved in modulation of such diverse physiological processes as response to genotoxic stress, regulation of the cell cycle, aging, and carcinogenesis. These new data provide a mechanistic link between deregulation of the circadian system and/or some of its core components and the development of various pathologies, suggesting novel strategies for the disease treatment and prevention. PMID- 18419308 TI - Sirtuins in aging and disease. AB - Sirtuin genes function as anti-aging genes in yeast, Caenorhabditis elegans, and Drosophila. The NAD requirement for sirtuin function indicates a link between aging and metabolism, and a boost in sirtuin activity may in part explain how calorie restriction extends life span. In mammals, one of the substrates of the SIR2 ortholog, SIRT1, is a regulator of mitochondrial biogenesis, PGC-1alpha. Indeed, the putative SIRT1 activator resveratrol has been shown to stimulate mitochondrial biogenesis and deliver health benefits in treated mice. I explore here how mitochondrial biogenesis may have beneficial effects on aging and, perhaps, diseases of aging. In particular, I speculate that SIRT1-mediated mitochondrial biogenesis may reduce the production of reactive oxygen species, a possible cause of aging, and offer two possible mechanisms for this effect. An understanding of how calorie restriction works may lead to novel drugs to combat diseases of aging. PMID- 18419309 TI - Identification of Caenorhabditis elegans genes regulating longevity using enhanced RNAi-sensitive strains. AB - A systematic genome-wide RNA interference screen was performed in the Caenorhabditis elegans lin-15b;eri-1 strain, which has an enhanced response to double-stranded RNA including the nervous system, to identify life-span regulatory factors. In total, 16,757 genes were examined, revealing 115 gene inactivations that extended life span. A more stringent longitudinal analysis revealed 18 gene inactivations that induced the greatest increase in life span (10-90%), all of which extended life span when inactivated either in eri-1 alone or in a second strain with an enhanced response to double-stranded RNA, eri-3. Most reduced the rate of aging, implying that animals aged more slowly. As was the case in previous studies, genes critical for metabolism caused the greatest extension of longevity. Extension of life span occurs through disparate mechanisms as increased resistance to thermal stress, oxidative damage, and decreased age pigment accumulation analysis of the 18 stronger positives failed to demonstrate a correlation between enhanced stress resistance and decreased lysosomal function. Consistently, aps-3 and lys-10, two genes annotated to have lysosomal functions, extended life span when inactivated without enhancing stress resistance. The results of this study reinforce the importance of metabolism, mitochondrial and lysosomal functions, genomic stability, and stress resistance on animal life-span determination. PMID- 18419310 TI - Circadian photoreception in vertebrates. AB - To be adaptively useful, internal circadian clocks must be entrained (synchronized) to daily rhythms in the external world. The entraining process adjusts the period of the internal clock to 24 hours and its phase to a value that determines the organism's temporal niche (e.g., diurnal and nocturnal). For most vertebrates, the dominant environmental synchronizer is light. All vertebrates employ specialized photoreceptor cells to perceive synchronizing light signals, but mammals and nonmammalian vertebrates do this differently. Mammals concentrate circadian photoreceptors in the retina, employing rods, cones, and a subset of retinal ganglion cells that are directly photosensitive and contain an unusual photopigment (melanopsin). Nonmammalian vertebrates use photoreceptors located deep in the brain and in the pineal gland as well as others in the retina. Such photoreceptor extravagance is difficult to explain. It seems likely that the different photoreceptor classes in this elaborate sensory system may have specialized roles in entrainment. There is some evidence that this is in fact the case. Furthermore, this nonvisual "circadian" photoreceptive system also controls acute behavioral responses to light (masking), pupillary constriction, and photoperiodic regulation of reproductive state. We review some of the early work on birds and describe new findings that indicate specific roles for retinal rods, cones, and photosensitive retinal ganglion cells in mammals. PMID- 18419311 TI - The lateral and dorsal neurons of Drosophila melanogaster: new insights about their morphology and function. AB - This chapter summarizes our present knowledge about the master clock of the fruit fly at the neuronal level. The clock is organized in distinct groups of interconnected pacemaker neurons with different functions. All of these neurons appear to communicate with one another in order to produce the species-specific activity rhythm, which is organized in morning (M) and evening (E) activity bouts. These two activity components are differentially influenced by distinct groups of pacemaker neurons reminiscent of the Pittendrigh-Daan dual oscillator model. In the original work (Grima et al. 2004; Stoleru et al. 2004), the ventrolateral (LN(v)) and dorsolateral (LN(d)) plus some dorsal groups (DN) of clock neurons have been defined as M and E cells, respectively. We further specify that the clock neurons belong to the M and E oscillators and define a more complex picture of the Drosophila brain clock. PMID- 18419313 TI - Inducible clocks: living in an unpredictable world. AB - All mammals have daily cycles of behavior (e.g., wake-sleep and feeding), and physiology (e.g., hormone secretion and body temperature). These cycles are typically entrained to the external light/dark cycle, but they can be altered dramatically under conditions of restricted food availability, changes in ambient temperature, or the presence of external stimuli such as predators. During the past 30 years, one of the best studied of these responses has been the entrainment of circadian rhythms to food availability. Experiments in rats and other rodents have provided evidence for a food-entrainable oscillator (FEO) in the mammalian circadian timing system (CTS). Until recently, however, very little was understood about the locus subserving the FEO or the functional interrelationship between the FEO and the master CTS pacemaker, the suprachiasmatic nucleus (SCN). We discuss here new data on the location of the FEO and suggest that it may involve an oscillator mechanism that is "induced" by starvation and refeeding. PMID- 18419314 TI - Suprachiasmatic nucleus clock time in the mammalian circadian system. AB - The integration of time from gene to system levels is an exciting feature of circadian biology. In mammals, clock cells in the suprachiasmatic nucleus (SCN) generate time by an autoregulatory transcription-(post)translational feedback loop. Clock activity in the SCN neurons is expressed as activity-dependent electric signals, which are coupled to those of other SCN neurons. The SCN spreads the time signals in a form of synchronized nerve impulses to central parasympathetic nuclei (e.g., dorsal motor nucleus of the vagus) and central sympathetic nuclei (e.g., intermediolateral cell column of the spinal cord). The vagal nerve innervates gastrointestinal and respiratory organs. Sympathetic signals to the adrenal gland are converted to hormonal (glucocorticoid) signals. Glucocorticoids released into the bloodstream bind to glucocorticoid receptors of peripheral organs, activate the mammalian Per1 gene in systemic cells, and reset the time of body clocks. Thus, the SCN-evoked time generated by specific genes localized to the SCN is converted to neuronal and hormonal signals and synchronizes the clocks in the whole body. PMID- 18419312 TI - Exploring spatiotemporal organization of SCN circuits. AB - Suprachiasmatic nucleus (SCN) neuroanatomy has been a subject of intense interest since the discovery of the SCN's function as a brain clock and subsequent studies revealing substantial heterogeneity of its component neurons. Understanding the network organization of the SCN has become increasingly relevant in the context of studies showing that its functional circuitry, evident in the spatial and temporal expression of clock genes, can be reorganized by inputs from the internal and external environment. Although multiple mechanisms have been proposed for coupling among SCN neurons, relatively little is known of the precise pattern of SCN circuitry. To explore SCN networks, we examine responses of the SCN to various photic conditions, using in vivo and in vitro studies with associated mathematical modeling to study spatiotemporal changes in SCN activity. We find an orderly and reproducible spatiotemporal pattern of oscillatory gene expression in the SCN, which requires the presence of the ventrolateral core region. Without the SCN core region, behavioral rhythmicity is abolished in vivo, whereas low-amplitude rhythmicity can be detected in SCN slices in vitro, but with loss of normal topographic organization. These studies reveal SCN circuit properties required to signal daily time. PMID- 18419315 TI - Molecular analysis of sleep: wake cycles in Drosophila. AB - Sleep is controlled by two major regulatory systems: a circadian system that drives it with a 24-hour periodicity and a home-ostatic system that ensures that adequate amounts of sleep are obtained. We are using the fruit fly Drosophila melanogaster to understand both types of regulation. With respect to circadian control, we have identified molecular mechanisms that are critical for the generation of a clock. Our recent efforts have focused on the analysis of posttranslational mechanisms, specifically the action of different phosphatases that control the phosphorylation and thereby the stability and/or nuclear localization of circadian clock proteins period (PER) and timeless (TIM). Resetting the clock in response to light is also mediated through posttranslational events that target TIM for degradation by the proteasome pathway; a recently identified ubiquitin ligase, jet lag (JET), is required for this response. Our understanding of the homeostatic control of sleep is in its early stages. We have found that mushroom bodies, which are a site of synaptic plasticity in the fly brain, are important for the regulation of sleep. In addition, through analysis of genes expressed under different behavioral states, we have identified some that are up-regulated during sleep deprivation. Thus, the Drosophila model allows the use of cellular and molecular approaches that should ultimately lead to a better understanding of sleep biology. PMID- 18419316 TI - Neurohormonal and neuromodulatory control of sleep in Drosophila. AB - The fruit fly Drosophila melanogaster has emerged in recent years as a tractable system for studying sleep. The sleep-wake dichotomy represents one of the principal transitions in global brain state, and neurohormones and neuromodulators are well known for their ability to change global brain states. Here, we describe studies of two brain systems that regulate sleep in Drosophila, the neurohormonal epidermal growth factor receptor system and the neuromodulatory dopaminergic system, each of which acts through a discrete anatomical locus in the dorsal brain. Both control systems display considerable mechanistic similarity to those in mammals, suggesting possible functional homologies. PMID- 18419317 TI - Molecular analysis of sleep. AB - Rest or sleep in all animal species constitutes a period of quiescence necessary for recovery from activity. Whether rest and activity observed in all organisms share a similar fundamental molecular basis with sleep and wakefulness in mammals has not yet been established. In addition and in contrast to the circadian system, strong evidence that sleep is regulated at the transcriptional level is lacking. Nevertheless, several studies indicate that single genesmay regulate some specific aspects of sleep. Efforts to better understand or confirm the role of known neurotransmission pathways in sleep-wake regulation using transgenic approaches resulted so far in only limited new insights. Recent gene expression profiling efforts in rats, mice, and fruit flies are promising and suggest that only a few gene categories are differentially regulated by behavioral state. How molecular analysis can help us to understand sleep is the focus of this chapter. PMID- 18419318 TI - Sleep and circadian rhythms in humans. AB - During the past 50 years, converging evidence reveals that the fundamental properties of the human circadian system are shared in common with those of other organisms. Concurrent data from multiple physiological rhythms in humans revealed that under some conditions, rhythms oscillated at different periods within the same individuals and led to the conclusion 30 years ago that the human circadian system was composed of multiple oscillators organized hierarchically; this inference has recently been confirmed using molecular techniques in species ranging from unicellular marine organisms to mammals. Although humans were once thought to be insensitive to the resetting effects of light, light is now recognized as the principal circadian synchronizer in humans, capable of eliciting weak (Type 1) or strong (Type 0) resetting, depending on stimulus strength and timing. Realization that circadian photoreception could be maintained in the absence of sight was first recognized in blind humans, as was the property of adaptation of the sensitivity of circadian photoreception to prior light history. In sighted humans, the intrinsic circadian period is very tightly distributed around approximately 24.2 hours and exhibits aftereffects of prior entrainment. Phase angle of entrainment is dependent on circadian period, at least in young adults. Circadian pacemakers in humans drive daily variations in many physiologic and behavioral variables, including circadian rhythms in alertness and sleep propensity. Under entrained conditions, these rhythms interact with homeostatic regulation of the sleep/wake cycle to determine the ability to sustain vigilance during the day and to sleep at night. Quantitative understanding of the fundamental properties of the multioscillator circadian system in humans and their interaction with sleep/wake homeostasis has many applications to health and disease, including the development of treatments for circadian rhythm and sleep disorders. PMID- 18419319 TI - Thermosensitive splicing of a clock gene and seasonal adaptation. AB - Similar to many diurnal animals, the daily distribution of activity in Drosophila exhibits a bimodal pattern with clock-controlled morning and evening peaks separated by a midday "siesta." In prior work, we showed that the thermosensitive splicing of a 3'-terminal intron in the RNA product from the Drosophila period (per) gene (dper) is critical for temperature-induced adjustments in the timing of evening activity. Cold temperatures enhance the splicing efficiency of this intron (termed dmpi8, Drosophila melanogaster per intron 8), an event that stimulates the daily accumulation of dper RNA and protein, leading to earlier evening activity. Conversely, warm temperatures attenuate dmpi8 splicing efficiency contributing to delayed evening activity, likely ensuring that flies avoid activity during the hot midday sun when they are at increased risk of desiccation. Here, we discuss the underlying molecular mechanisms governing the thermosensitive splicing of dmpi8 and how it contributes to seasonal changes in the daily activity patterns of Drosophila. On a broader perspective, RNA-RNA interactions likely have fundamental roles in the thermal adaptation of life forms to the daily and seasonal changes in temperature. PMID- 18419320 TI - Endogenous circannual clock and HP complex in a hibernation control system. AB - Hibernation in mammals is a mysterious biological phenomenon that appears on a seasonal basis for surviving a potentially lethal low body temperature (Tb) near 0 degrees C and protecting organisms from various diseases and harmful events during hibernation. The exact mechanism by which such a unique ability is seasonally developed is still unknown. On the basis of our previous finding that the source of calcium ions for excitation-contraction coupling in myocardium of chipmunks, a rodent hibernator, is seasonally modulated for hibernation, the liver-derived hibernation-specific protein (HP) complex was discovered. Recently, the HP complex was identified as a promising candidate hormone that carries a hibernation signal to the brain independently of Tb and environmental changes for developing a capacity for tolerating low Tb. This finding will promote new approaches to understanding biological hibernation systems, including a circannual clock and its signaling pathway between the brain and the periphery. A new definition of hibernation and a possible model of a hibernation control system are proposed. PMID- 18419321 TI - On the chronobiology of cohabitation. AB - Social regulation of animal circadian rhythms may enable individuals in a population to temporally synchronize or segregate their activities within the community. Relatively little is known about the mechanisms for such interindividual temporal adaptations or how the circadian system might be involved. The literature suggests that actual prolonged cohabitation might lead to robust effects on the rhythmicity of cohoused individuals but that these effects are not easily reproduced by indirect or pulsatile social contacts. We have begun to study the conditions under which such cohabitation effects might be revealed in the laboratory, and we present and discuss initial data that cohousing pairs of golden hamsters can result in a persistent change in the free running circadian period of one of the two hamsters of the pair. We believe that analyzing the societal level of temporal organization, and ultimately dissecting its underlying mechanisms, will enrich our understanding of the circadian clock and its role in establishing ecological communities. PMID- 18419322 TI - Melatonin and human chronobiology. AB - With the development of accurate and sensitive assays for measuring melatonin in plasma and saliva, it has been possible to advance our understanding of human chronobiology. In particular, the dim light melatonin onset (DLMO) is expected to have an increasingly important role in the diagnosis of circadian phase disorders and their treatment with appropriately timed bright light exposure and/or low dose melatonin administration. The phase angle difference (PAD) between DLMO and mid-sleep can be used as a marker for internal circadian alignment and may also be used to differentiate individuals who are phase advanced from those who are phase delayed (a long interval indicates the former and a short interval indicates the latter). To provide a corrective phase delay, light exposure should be scheduled in the evening and melatonin should be administered in the morning. To provide a corrective phase advance, light exposure should be scheduled in the morning and melatonin should be administered in the afternoon/evening. The study of patients with seasonal affective disorder (SAD), as well as individuals who are totally blind, has resulted in several findings of interest to basic scientists, as well as psychiatrists and sleep specialists. PMID- 18419323 TI - Role for the Clock gene in bipolar disorder. AB - Nearly all patients with bipolar disorder have severely disrupted circadian rhythms. Treatment with mood stabilizers can restore these daily rhythms, and this is correlated with patient recovery. However, it is still uncertain whether clock abnormalities are the cause of bipolar disorder or if these rhythm disruptions are secondary to alterations in other circuits. Furthermore, the mechanism by which the circadian clock might influence mood is still unclear. With cloning and characterization of the circadian genes and recent advances in molecular biology, we are starting to understand this strong association between circadian rhythms and bipolar disorder. Recent human genetic and mouse behavioral studies indicate that the Clock gene is particularly relevant in the mood disruptions associated with this disorder. Furthermore, it appears that Clock expression outside of the central pacemaker of the suprachiasmatic nucleus (SCN) is involved in mood regulation. In this chapter, the evidence linking circadian rhythms, the Clock gene, and bipolar disorder is discussed, along with the possible biology that underlies this connection. PMID- 18419324 TI - The possible interplay of synaptic and clock genes in autism spectrum disorders. AB - Autism spectrum disorders (ASD) are complex neurodevelopmental conditions characterized by deficits in social communication, absence or delay in language, and stereotyped and repetitive behaviors. Results from genetic studies reveal one pathway associated with susceptibility to ASD, which includes the synaptic cell adhesion molecules NLGN3, NLGN4, and NRXN1 and a postsynaptic scaffolding protein SHANK3. This protein complex is crucial for the maintenance of functional synapses as well as the adequate balance between neuronal excitation and inhibition. Among the factors that could modulate this pathway are the genes controlling circadian rhythms. Indeed, sleep disorders and low melatonin levels are frequently observed in ASD. In this context, an alteration of both this synaptic pathway and the setting of the clock would greatly increase the risk of ASD. In this chapter, I report genetic and neurobiological findings that highlight the major role of synaptic and clock genes in the susceptibility to ASD. On the basis of these lines of evidence, I propose that future studies of ASD should investigate the circadian modulation of synaptic function as a focus for functional analyses and the development of new therapeutic strategies. PMID- 18419325 TI - Circadian clocks: 50 years on. AB - Since the first Cold Spring Harbor meeting on "Biological Clocks" in 1960, the field has progressed from the study of a fascinating but esoteric set of phenomena of interest primarily to a relatively small group of prescient biologists to become recognized as defining a centrally important aspect of biological organization. This change is the consequence of a profound increase in understanding of the mechanisms that generate and control circadian rhythmicity, coupled with the realization that circadian temporal organization is an important component of much of what most organisms do. As such, it impinges on human health, agriculture, and biological conservation, as well as on many more basic aspects of biology at every level. Many of the seminal discoveries of the last 47 years were presented and discussed at this exciting meeting. PMID- 18419328 TI - Patient tolerance during cystoscopy: a randomized study comparing lidocaine hydrochloride gel and dimethyl sulfoxide with lidocaine. AB - PURPOSE: Cystoscopy is one of the most common examinations in urologic outpatient clinics. Various anesthetic approaches have been used to make cystoscopy more tolerable for patients. The aim of the present prospective randomized study was to evaluate the efficacy of lidocaine hydrochloride gel compared to dimethyl sulfoxide (DMSO) with lidocaine in rigid cystoscopy. MATERIALS AND METHODS: Male patients requiring 17F rigid cystoscopy were eligible for inclusion in this study. A total of 140 patients were divided into two groups: group 1 (n=70) received approximately 11 mL of 2% lidocaine gel intraurethrally, while in group 2 (n=70) approximately 10 mL of 40% DMSO with an amount of lidocaine equal to that in the lidocaine gel was smeared around the scope and external urethral meatus. A penile clamp was placed for 15 minutes and 5 minutes in group 1 and group 2, respectively. Immediately after cystoscopic examination pain was scored on a 10-cm visual analog scale. RESULTS: The mean pain scores after the procedure for group 1 and group 2 were 3.9+/-1.1 and 2.1+/-1.0, respectively. The pain scores were significantly lower for group 2 than for group 1 (P=0.015). No patients needed additional anesthetic agents or sedatives due to insufficient analgesia, and there were no serious side effects in either group. CONCLUSIONS: Our study has shown that DMSO with lidocaine gel causes significantly less delivery discomfort in the male urethra than lidocaine hydrochloride gel. The advantages of DMSO with lidocaine are the mixture takes less time to act and had lower pain scores. PMID- 18419330 TI - Laparoscopic radical cystectomy with orthotopic ileal neobladder: a report of 85 cases. AB - PURPOSE: The preliminary results of laparoscopic radical cystectomy in 85 patients are presented in this study. The functional and oncologic outcomes of this procedure in these patients are discussed. PATIENTS AND METHODS: Between December 2002 and May 2006, we performed 85 laparoscopic radical cystectomies with orthotopic ileal neobladder for bladder cancer in 77 men and 8 women. A 5 port transperitoneal approach was applied. The standard bilateral pelvic lymphadenectomy was performed first, then radical cystectomy was completed laparoscopically. The construction of the ileal neobladder and the anastomosis of ureter-neobladder were performed extracorporeally. The neobladder was anastomosed to the urethral stump under laparoscopy. A nerve-sparing procedure was performed for eight patients. RESULTS: The median operative time was 320 min, and the median blood loss was 280 mL. Conversion to open surgery was not necessary in any of the patients. The average time to oral intake after operation was 3.9 days. There were no perioperative mortalities. The complication rate was 14.1% (12/85), including such complications as three uretero-pouch anastomotic strictures, one vesicourethral anastomotic stricture, one pouch-vaginal fistula, one colonic pouch fistula, one ileo-pouch fistula, three ileus, one pneumonia, and one pyelonephritis. The daytime continence rate was 91.2%, and the nighttime continence rate was 82.4% at 6 months postoperatively. The neobladder capacity was about 343 mL. Surgical margins were tumor free for all patients. Of the eight patients who underwent a nerve-sparing procedure, four patients had potency for intercourse. During a follow-up period of 1 to 41 months (average 21.3 months), three patients had local recurrence, one patient had trocar site seeding, and five patients had distant metastasis, of whom four died. CONCLUSIONS: Laparoscopic radical cystectomy with extracorporeal formation of a neobladder is a feasible procedure with low morbidity and acceptable neobladder function. Long term follow-up is needed to confirm the oncologic outcomes. PMID- 18419331 TI - Management of bladder stones with pneumatic lithotripsy using a ureteroscope in children. AB - PURPOSE: To evaluate the effectiveness and safety of pneumatic lithotripsy by using a ureteroscope to treat bladder stones in children. PATIENTS AND METHODS: Twenty-seven boys presenting with bladder stones underwent transurethral cystolithotripsy. The indication for transurethral cystolithotripsy was stone size 0.05). No patient (0%) in the BL-SNG group ever achieved RTB-SF status at any time point. PSMs were observed in 37% (10/27) of all patients. BCR occurred in nine patients (33.3%), seven of whom had PSM (78%); treatment failure occurred within 6 months of surgery, necessitating androgen deprivation therapy. CONCLUSION: Despite optimism regarding SNG, long-term functional outcomes have been disappointing, particularly for BL nerve interposition. UL-SNG functional outcomes do not appear to improve outcomes when compared with men with UL nerve preservation. With the greater risk of PSM and BCR in patients who are considered candidates for SNG, newer treatment modalities are needed to cure their disease while preserving SF. PMID- 18419334 TI - Robotic radical prostatectomy: assurance of water-tight vesicourethral anastomotic closure with the Lapra-Ty clip. AB - Most surgeons use a continuous running suture technique during robot-assisted radical prostatectomy followed by knot tying. Because of lack of haptic feedback and the potential for suboptimal suture tension after knot tying, the anastomotic closure may not be water-tight. Use of the Lapra-Ty clip (LTc) during anastomotic reconstruction offers the surgeon the ability to readjust suture tension as well as salvage torn sutures. Here we discuss our current anastomotic technique and highlight specific situations in which LTc use may prove beneficial. PMID- 18419335 TI - Management of ureteropelvic junction obstruction. PMID- 18419336 TI - Conventional high pressure versus newly developed continuous-flow ureterorenoscope: urodynamic pressure evaluation of the renal pelvis and flow capacity. AB - PURPOSE: We evaluated the pressure and flow relation of a newly developed continuous-flow ureterorenoscope (URS) in comparison with a common ureterorenoscope in an ex-vivo urinary tract model. MATERIALS AND METHODS: Ureterorenoscopies were performed with the newly developed 10.5F continuous-flow URS with separate inflow and outflow channel and a conventional 10.5F URS with a combined inflow and outflow channel. The ex-vivo model consisted of complete urinary tracts of domestic pigs obtained freshly from the slaughterhouse. Both instruments were used in five urinary tracts, and six ureterorenoscopies were performed in each urinary tract. The pressure in the renal pelvis (RP) was measured during each procedure. Height of the irrigation solution above renal level and flow capacity were also documented. RESULTS: The conventional URS showed a correlation of intrapelvic pressure and the height of the irrigation solution above renal level rising from 20+/-3.7 cm H(2)O at a solution level of 20 cm to a plateau pressure of 40+/-3.3 cm H(2)O with a distinct renal influx at a level of 50 cm. The maximum flow capacity at a solution level of 20 cm was 0.2 mL/min rising to a flow capacity of 0.5 mL/min at 40 cm above renal level. The maximum flow capacity for the continuous-flow URS was about 100 times higher, rising from 20 mL/min at a solution level of 20 cm to 70 mL/min at 40 cm above renal level. The intrapelvic pressure was 15+/-2.1 cm H(2)O at a solution level of 20 cm and did not exceed the physiologic renal pressure of 20 cm H(2)O even if the irrigation solution was at a height of 100 cm above renal level. CONCLUSION: The newly developed continuous-flow URS provides a 100 times higher flow capacity while simultaneously preserving the physiologic pressure in the RP compared with the conventional URS. These characteristics will improve visibility and reduce retrograde stone manipulation, operative time, and complications under clinical conditions. PMID- 18419337 TI - Pure natural orifice transluminal endoscopic surgery (NOTES) nephrectomy using standard laparoscopic instruments in the porcine model. AB - PURPOSE: Natural Orifice Transluminal Endoscopic Surgery (NOTES) is an emerging technology that uses endoscopic instruments passed into the peritoneal cavity through hollow viscera to perform surgical procedures without the use of abdominal incisions. There are, however, limitations regarding the equipment available to simulate traditional surgery. The aim of this study was to determine the feasibility of a pure NOTES nephrectomy by using standard laparoscopic instruments through a modified transvaginal trocar. MATERIALS AND METHODS: One 40 kg female swine underwent transgastric peritoneoscopy. Transgastric endoscopic visualization guided the introduction of a second transvaginal endoscope through a novel laparoscopic trocar/endoscopic overtube device. The retroflexed transgastric endoscope provided triangulated visualization as standard endoscopic instruments provided retraction, which allowed dissection of the kidney with standard laparoscopic instruments through our modified transvaginal trocar device. Each renal hilum, artery, vein, and ureter was dissected and divided with a transvaginal laparoscopic stapler. RESULTS: Transgastric and transvaginal NOTES accesses were easily achieved, and bilateral nephrectomies were performed. Completion of peritoneoscopy revealed complete hemostasis and identification of ligated ureters and hilar vessels. Total operative time was 40 and 20 minutes for the right and left kidney, respectively. One kidney was captured with a laparoscopic retrieval sac and removed intact through the vaginal defect. CONCLUSIONS: Pure NOTES nephrectomies are technically feasible in the porcine model by using standard laparoscopic instruments. Survival studies are necessary to determine the long-term complications and physiologic implications of NOTES nephrectomy. The development of innovative NOTES access trocars may allow for an increased armamentarium of NOTES instruments. PMID- 18419338 TI - Case report: percutaneous fibrin glue injection for urine leakage in laparoscopic partial nephrectomy. AB - As laparoscopic techniques have advanced in the recent decade, laparoscopic partial nephrectomy (LPN) has been performed in patients with renal-cell carcinoma (RCC). Until recently, however, it has been a challenging procedure to perform because of its technical difficulty. We treated a 71-year-old man with a single kidney and RCC who subsequently underwent LPN. Urine leakage was detected postoperatively and continued for 30 days. It was managed with a percutaneous fibrin glue injection. Our report shows the therapeutic feasibility of fibrin glue for urine leakage, a complication of LPN. PMID- 18419339 TI - Genotypic and environmental effects on flight activity and oviposition in the Glanville fritillary butterfly. AB - Adverse environmental conditions constrain active flight and thereby limit reproduction in most insects. Butterflies have evolved various adaptations in order to thermoregulate, allowing females to search for nectar and to oviposit under unfavorable thermal conditions. We studied experimentally and with observational data the effect of low ambient temperatures experienced in the morning on the timing of oviposition and clutch size in the Glanville fritillary butterfly (Melitaea cinxia). Comparisons were made between individuals with different forms of the gene Pgi, encoding the glycolytic enzyme phosphoglucose isomerase, since naturally segregating variation at Pgi is known to be correlated with flight metabolic rate, flight performance, and fecundity. Experiencing low temperature in the morning delayed the initiation of oviposition and decreased clutch size. We used a thermal image camera to measure the thoracic surface temperature of butterflies immediately after voluntary flight. Single nucleotide polymorphism at Pgi was associated with thoracic temperature at low ambient temperatures. This has consequences for reproduction because females that are able to fly at lower ambient temperatures generally initiate oviposition earlier in the afternoon, when the environmental conditions are most favorable and the average egg clutch size is generally largest. These results suggest that variation in physiological and molecular capacity to sustain active flight at low ambient temperature has significant fitness-related consequences in insects. PMID- 18419340 TI - Evolution of avian plumage color in a tetrahedral color space: a phylogenetic analysis of new world buntings. AB - We use a tetrahedral color space to describe and analyze male plumage color variation and evolution in a clade of New World buntings--Cyanocompsa and Passerina (Aves: Cardinalidae). The Goldsmith color space models the relative stimulation of the four retinal cones, using the integrals of the product of plumage reflectance spectra and cone sensitivity functions. A color is represented as a vector defined by the relative stimulation of the four cone types--ultraviolet, blue, green, and red. Color vectors are plotted in a tetrahedral, or quaternary, plot with the achromatic point at the origin and the ultraviolet/violet channel along the Z-axis. Each color vector is specified by the spherical coordinates theta, phi, and r. Hue is given by the angles theta and phi. Chroma is given by the magnitude of r, the distance from the achromatic origin. Color vectors of all distinct patches in a plumage characterize the plumage color phenotype. We describe the variation in color space occupancy of male bunting plumages, using various measures of color contrast, hue contrast and diversity, and chroma. Comparative phylogenetic analyses using linear parsimony (in MacClade) and generalized least squares (GLS) models (in CONTINUOUS) with a molecular phylogeny of the group document that plumage color evolution in the clade has been very dynamic. The single best-fit GLS evolutionary model of plumage color variation over the entire clade is a directional change model with no phylogenetic correlation among species. However, phylogenetic innovations in feather color production mechanisms--derived pheomelanin and carotenoid expression in two lineages--created new opportunities to colonize novel areas of color space and fostered the explosive differentiation in plumage color. Comparison of the tetrahedral color space of Goldsmith with that of Endler and Mielke demonstrates that both provide essentially identical results. Evolution of avian ultraviolet/violet opsin sensitivity in relation to chromatic experience is discussed. PMID- 18419341 TI - Evolution in metacommunities: on the relative importance of species sorting and monopolization in structuring communities. AB - Abstract: Adaptive evolution within species and community assembly involving multiple species are both affected by dispersal and spatiotemporal environmental variation and may thus interact with each other. We examined this interaction in a simple three-patch metacommunity and found that these two processes produce very different associations between species composition and local environment. In most conditions, we find a pattern we call "species sorting," wherein local adaptation by resident species cannot prevent invasions by other preadapted species as environmental conditions change (strong association between local environmental conditions and local community composition). When dispersal rates are very low relative to the other two rates, local adaptation by resident species predominates, leading to strong priority effects that prevent successful colonization by other species that would have been well adapted, a pattern we call "local monopolization." When dispersal and evolutionary rates are both very high, we find that an evolving species outcompetes other species in all patches, a pattern we call "global monopolization." When environmental oscillations are very frequent, local monopolization predominates. Our findings indicate that there can be strong modification of community assembly by local adaptive processes and that these depend strongly on the relative rates of evolution, dispersal, and environmental change. PMID- 18419343 TI - Common nonsynonymous polymorphisms in the NOD2 gene are associated with resistance or susceptibility to tuberculosis disease in African Americans. AB - Pattern-recognition receptors (PRRs) play a key role in innate immunity against intracellular bacteria. NOD2 is one of the PRRs that contribute to the immune response to Mycobacterium tuberculosis infection. We sequenced coding regions of the NOD2 gene in 377 African Americans with tuberculosis (TB) disease and 187 ethnically matched control subjects. Three common nonsynonymous single-nucleotide polymorphisms--Pro268Ser, Arg702Trp, and Ala725Gly--demonstrated significant associations with TB disease. This finding may contribute to the future development of immunotherapy and immunoprophylaxis for TB disease. PMID- 18419342 TI - Superantigen profile of Staphylococcus aureus isolates from patients with steroid resistant atopic dermatitis. AB - BACKGROUND: Superantigens induce skin inflammatory responses in atopic dermatitis, which is commonly associated with Staphylococcus aureus infection. T cells activated in vitro by superantigens become steroid resistant. The objective was to assess the superantigen profiles of S. aureus isolates from patients with steroid-resistant atopic dermatitis. METHODS: We compared the superantigen production capability of S. aureus isolates from 78 patients with steroid resistant atopic dermatitis (group 1) with that of 30 vaginal isolates from healthy women (group 2) and 22 isolates from a general population of patients with atopic dermatitis (group 3). Polymerase chain reaction with primers for superantigens, combined with selected antibody testing, was used to analyze the presence of toxic shock syndrome toxin 1, staphylococcal enterotoxins, and enterotoxin-like superantigens. RESULTS: S. aureus isolates from group 1 had a statistically significant difference in superantigen profile, compared with the profiles of group 2 and group 3 isolates. Group 2 isolates were similar in profile to group 3 isolates, with 4 and 5 superantigens per isolate, respectively. In contrast, group 1 isolates produced a mean of 8 superantigens each (P<<.001, for comparison with group 2 or group 3). These group 1 isolates were more likely to produce the 3 major toxic shock syndrome-associated superantigens (toxic shock syndrome toxin 1, staphylococcal enterotoxin B, and staphylococcal enterotoxin C) and to produce unusual combinations of superantigens (e.g., toxic shock syndrome toxin 1 and staphylococcal enterotoxin B). CONCLUSIONS: S. aureus isolates from patients with steroid-resistant atopic dermatitis appear to be selected on the basis of greater production of superantigens, compared with that of isolates from control groups. Superantigens may offer selective advantages for colonization of patients. PMID- 18419344 TI - Impact of HIV on host-virus interactions during early hepatitis C virus infection. AB - BACKGROUND: Human immunodeficiency virus (HIV) may influence the outcome and natural history of hepatitis C virus (HCV) infection through an impact on acute HCV-specific T cell responses. METHODS: Fifty-five HIV-positive males with acute HCV infection were identified; monoinfected individuals (n = 8) were used for peripheral blood mononuclear cell comparison. In 14 coinfected and 8 HCV monoinfected patients, HCV-specific T cell responses against a range of HCV antigens were assessed using interferon (IFN)-gamma enzyme-linked immunospot (ELISpot) and proliferation assays. E1/E2 region genetic diversity and the selection pressure on the virus were measured in 8 coinfected patients by use of cloned sequences over time. RESULTS: HCV persisted in 52 (95%) coinfected individuals. HCV/HIV coinfection significantly reduced IFN-gamma ELISpot responses versus those in HCV-monoinfected individuals, especially against nonstructural proteins (1/10 vs. 5/8; P = .008). In coinfected patients, increased HCV genetic diversity was observed between the first and subsequent time points, with no evidence for positive selection in the E1/E2 region sequenced. CONCLUSION: HIV coinfection is associated with increased rates of HCV persistence and a lack of critical CD4 T cell responses, with no evidence of immune selection pressure during early HCV infection. Loss of key cellular immune responses against HCV during acute disease may contribute to the failure of early host control of HCV in HCV/HIV-coinfected patients. PMID- 18419345 TI - Immune responses to mumps vaccine in adults who were vaccinated in childhood. AB - BACKGROUND: In a mumps outbreak in the United States, many infected individuals were adults who had received 2 doses of mumps vaccine. The persistence of cellular immunity to mumps vaccine has not been defined. METHODS: This was an observational, nonrandomized cohort study evaluating cell-mediated and humoral immunity to mumps in 10 vaccinated and 10 naturally immune adults. Mumps-specific T cell activation and interferon (IFN)-gamma production were measured using lymphoproliferative and flow cytometry assays, and mumps immunoglobulin (Ig) G was measured using enzyme-linked immunosorbent assay. RESULTS: T cell immunity to mumps was high in both groups; 70% of vaccinated and 80% of naturally immune individuals had a positive (> or =3) stimulation index (SI) (P = 1.0). The mean percentages of mumps-specific CD4+ T cells that expressed CD69 and produced IFN gamma were equivalent in the 2 groups: 0.06% and 0.12%, respectively (P = .11). The mean SIs in the groups were also equivalent, although IFN-gamma concentrations from cultures stimulated with mumps antigen were higher in naturally immune adults than in vaccinated adults (P < or = .01). All adults were positive for mumps IgG. CONCLUSION: T and B cell immunity to mumps was detected in adults at least 10 years after immunization. Except for IFN-gamma release, responses in vaccinated adults paralleled those observed in naturally immune individuals. PMID- 18419346 TI - Long-term persistence of mumps antibody after receipt of 2 measles-mumps-rubella (MMR) vaccinations and antibody response after a third MMR vaccination among a university population. AB - BACKGROUND: High attack rates among vaccinated young adults reported during the 2006 mumps outbreak in the United States heightened concerns regarding mumps vaccine failure. METHODS: Serum specimens from university students and staff were tested for mumps immunoglobulin (Ig) G by enzyme immunoassay (EIA). A subset of participants vaccinated for < or =5 years and > or =15 years were tested by neutralizing antibody (NA) assay. Persons seronegative by EIA were offered a third dose of measles-mumps-rubella vaccine (MMR3), and serum specimens were obtained 7-10 days and 2-3 months after its administration. RESULTS: Overall, 94% (95% confidence interval [CI], 91%-96%) of the 440 participants were seropositive. No differences existed in seropositivity rates by sex, age, age at receipt of the second dose of MMR vaccine (MMR2), or time since receipt of MMR2 (P = .568). The geometric mean titer (GMT) of NA among persons vaccinated with MMR2 during the previous 1-5 years was 97 (95% CI, 64-148), whereas, among those vaccinated > or =15 years before blood collection, the GMT was 58 (95% CI, 44-76) (P = .065). After MMR3, 82% (14/17) and 91% (10/11) seroconverted in 7-10 days and 2-3 months, respectively. CONCLUSIONS: Lower levels of NA observed among persons who received MMR2 > or =15 years ago demonstrates antibody decay over time. MMR3 vaccination of most seronegative persons marked the capacity to mount an anamnestic response. PMID- 18419347 TI - Molecular markers of resistance to sulfadoxine-pyrimethamine during intermittent preventive treatment for malaria in Mozambican infants. AB - BACKGROUND: Intermittent preventive treatment in infants (IPTi) with sulfadoxine pyrimethamine (SP) is a potential malaria control strategy. There is concern about the impact that increasing in vivo resistance to SP has on the efficacy of IPTi, as well as about the potential contribution of IPTi to increases in resistance. METHODS: We compared the frequency of clinical episodes of malaria caused by P. falciparum parasites with mutations in dhfr and dhps among sick children who received SP or placebo in the context of a randomized, double-blind, placebo-controlled IPTi trial in Mozambique. RESULTS: Half of the children who received placebo harbored quintuple-pure mutant parasites. Nevertheless, the protective efficacy of IPTi within the 35 days after the third dose was 70.8% (95% confidence interval [CI], 40.7%-85.6%). Between month 2 after the third IPTi dose and the end of the follow-up period, children receiving SP harbored more dhps codon 437 mixed infections (odds ratio [OR], 10.56 [95% CI, 1.30-86.14]) and fewer dhps double-pure mutant parasites (OR, 0.43 [95% CI, 0.22-0.84]) than did placebo recipients. CONCLUSIONS: IPTi appears to be associated with some changes in the prevalence of genotypes involved in SP resistance. In the face of a high prevalence of quintuple-mutant parasites, SP exhibited a high level of efficacy in the prevention of new episodes of malaria in infants. PMID- 18419348 TI - Human immunodeficiency virus-associated lipoatrophy: letting the genome out of the bottle. PMID- 18419349 TI - Varicella-zoster virus-specific immune responses in elderly recipients of a herpes zoster vaccine. AB - BACKGROUND: A double-blind, placebo-controlled trial that involved 38,546 subjects > or =60 years old demonstrated efficacy of a high-potency live attenuated Oka/Merck varicella-zoster virus (VZV) vaccine. The trial included an immunology substudy to determine the relationship of VZV-specific immune responses to vaccination and clinical outcome. METHODS: The immunology substudy enrolled 1395 subjects at 2 sites where blood samples obtained prior to vaccination, at 6 weeks after vaccination, and at 1, 2, and 3 years thereafter were tested for VZV-specific cell-mediated immunity (VZV-CMI) by gamma-interferon ELISPOT and responder cell frequency assays and for VZV antibody by glycoprotein ELISA. RESULTS: VZV-CMI and VZV antibodies were significantly increased in vaccine recipients at 6 weeks after vaccination. The vaccine-induced increases in VZV-CMI persisted during the 3 years of follow-up, although their magnitude decreased over time. The magnitude of these VZV-specific immune responses was greater in subjects 60-69 years old than in subjects > or =70 years old. CONCLUSIONS: The zoster vaccine induced a significant increase in VZV-CMI and VZV antibody. The magnitude and duration of the boost in VZV-CMI in vaccine recipients and the relationship of this boost to age paralleled the clinical effects of the vaccine observed during the efficacy trial. These findings support the hypothesis that boosting VZV-CMI protects older adults against herpes zoster and postherpetic neuralgia. PMID- 18419351 TI - Prevalence of asymptomatic Tropheryma whipplei carriage among humans and nonhuman primates. AB - BACKGROUND: The reservoir of the agent of Whipple disease is unknown. Asymptomatic carriage of Tropheryma whipplei in human stool and saliva is controversial. METHODS: Stools and saliva specimens from 231 workers at a sewage treatment facility and from 10 patients with Whipple disease, stool specimens from 102 healthy people, and stool specimens from 127 monkeys or apes were tested for T. whipplei DNA by a quantitative real-time polymerase chain reaction with probe detection. Genotyping and culture of T. whipplei-positive samples were performed. RESULTS: Asymptomatic carriage in stool was found in humans (ranging from a prevalence of 4% in the control group to 12% among a subgroup of sewer workers) but not in monkeys and apes. The T. whipplei load in stool was significantly lower in carriers than in patients with Whipple disease (P < .001). There was a significant prevalence gradient associated with employment responsibilities at the sewage treatment facility: workers who cleaned the underground portion of the sewers were more likely than other workers to carry T. whipplei in stool. Seven of 9 sewer workers tested positive 8 months later. Patients with Whipple disease were significantly more likely to have T. whipplei positive saliva specimens (P = .005) and had a significantly greater T. whipplei load in saliva (P = .015), compared with asymptomatic stool carriers from the sewage facility. All non-stool carriers had T. whipplei-negative saliva specimens. T. whipplei strains were heterogeneic among sewer workers but identical within individual workers. CONCLUSION: Chronic asymptomatic carriage of T. whipplei occurs in humans. Bacterial loads are lower in asymptomatic carriers, and the prevalence of carriage increases with exposure to sewage. PMID- 18419352 TI - Resolution of hepatitis C virus-induced steatosis improves tolerability of antiretroviral drugs associated with hepatotoxicity in an HIV-infected individual. PMID- 18419350 TI - Hemochromatosis gene polymorphisms, mitochondrial haplogroups, and peripheral lipoatrophy during antiretroviral therapy. AB - BACKGROUND: Antiretroviral therapy (ART)-associated lipoatrophy involves mitochondrial dysfunction. Iron metabolism impacts mitochondrial function and oxidative stress. Mitochondrial haplogroups and hemochromatosis gene (HFE) polymorphisms have been associated with ART-induced neuropathy. We assessed relationships between these variants and lipoatrophy. METHODS: The AIDS Clinical Trials Group 384 study randomized ART-naive individuals to receive didanosine stavudine or zidovudine-lamivudine, combined with efavirenz and/or nelfinavir. Substudy A5005s evaluated fat distribution by dual-energy X-ray absorptiometry (DEXA). We characterized HFE polymorphisms 845G>A and 187C>G and European mitochondrial haplogroups in A5005s participants who consented to genetic analyses. RESULTS: Among 96 participants (58% were white, and 10% were female) with baseline and 48 or 64 week DEXA data, the median limb fat change was -8.8% (interquartile range, -28.7% to +15.6%). HFE 187C/G heterozygotes (n = 23) had less limb fat loss than 187C/C homozygotes (n = 71) (+6.1% vs. -12.5%; P = .02) and were less likely to develop lipoatrophy after adjustment for age, sex, race, and ART randomization (odds ratio, 0.31; 95% confidence interval, 0.10-0.95; P = .04). Among non-Hispanic white participants, median limb fat change was +26.1% among 5 participants with mitochondrial haplogroup J, compared with -9.7% among 49 participants with other mitochondrial haplogroups (P = .07). CONCLUSIONS: HFE 187C>G and, possibly, mitochondrial haplogroup J gave relative protection against lipoatrophy during ART in A5005s. These associations should be replicated in other studies. PMID- 18419355 TI - Disease progression and fatal outcomes in HIV-infected patients during interruption of antiretroviral treatment. PMID- 18419356 TI - Human papillomavirus (HPV) genotype 84 infection of the male genitalia: further evidence for HPV tissue tropism? PMID- 18419359 TI - Challenges in cost-effectiveness analyses of hospital interventions to improve quality. PMID- 18419360 TI - Factors associated with acquisition of vancomycin-resistant enterococci (VRE) in roommate contacts of patients colonized or infected with VRE in a tertiary care hospital. AB - OBJECTIVE: Most nosocomial acquistion of vancomycin-resistant enterococci (VRE) is due to cross-transmission. We sought to identify risk factors for acquisition of VRE by roommates of patients colonized or infected with VRE. DESIGN: Retrospective cohort study. SETTING: A 472-bed tertiary care teaching hospital. METHODS: All patients who shared a room with a patient colonized or infected with VRE at our hospital between January 1, 1999 and December 31, 2006 were identified. These roommates of VRE-positive patients were screened by rectal swab culture on days 2, 5, and 7 after the last exposure to the index patient. Chart reviews were performed to identify risk factors for VRE colonization in these roommates. RESULTS: Eighty-eight roommates of patients colonized or infected with VRE were identified. Of the 38 roommates with complete follow-up, 8 (21%) became colonized with VRE. These 8 roommates were older (median, 87.5 vs 62.5 years of age; P = .001), had longer duration of room exposure (median, 8.5 vs 4 days; P = .002), and were more likely to have a urinary catheter (odds ratio [OR], 16 [95% confidence interval {CI}, 1.7-152]; P = .005), an elevated serum creatinine level (OR, 17 [95% CI, 1.4-196]; P = .02), low serum albumin level (OR, 9.9 [95% CI, 1.3-113]; P = .01), and recent third-generation cephalosporin use (OR, 8.3 [95% CI, 1.5-47]; P = .02). CONCLUSION: Roommates of patients identified as colonized or infected with VRE are at substantial risk of becoming colonized, with the degree of risk increasing in older and more frail patients. VRE control programs should pay particular attention to such patients. PMID- 18419361 TI - Risk of vancomycin-resistant Enterococcus (VRE) bloodstream infection among patients colonized with VRE. AB - BACKGROUND: Colonization with vancomycin-resistant Enterococcus (VRE) is a risk factor for subsequent VRE bloodstream infection (BSI); however, risk factors for BSI among colonized patients have not been adequately described. We sought to determine the proportion of VRE-colonized patients who subsequently develop VRE BSI and to identify risk factors for VRE BSI among these patients. METHODS: Records of 768 patients colonized with VRE from January 2002 through June 2005 were reviewed. The proportion of patients who developed VRE BSI was calculated, and the characteristics of these patients were compared, in a 2:1 ratio, with those of patients who did not develop VRE BSI. To identify risk factors for VRE BSI and for death, we used univariate logistic regression analysis and then multivariate logistic regression analysis. Using pulsed-field gel electrophoresis (PFGE), we compared the isolate recovered when the patient was colonized and the isolate recovered when the patient developed VRE BSI. RESULTS: Of the 768 patients colonized with VRE, 31 (4.0%) developed VRE BSI. Multivariate analysis identified the following independent risk factors for developing VRE BSI: infection of an additional body site other than blood (adjusted odds ratio [aOR], 3.9; P = .04), admission to the hospital from a long-term care facility (aOR, 12.6; P = .04), and receipt of vancomycin (aOR, 10.6; P = .001). The independent risk factors for death among patients colonized with VRE were immunosuppression (aOR, 12.9; P = .001) and VRE BSI (aOR, 9.1; P = .002). Of the 31 patients who developed VRE BSI, 23 (74%) had a pair of isolates representing VRE colonization and VRE BSI. For 19 (83%) of these 23 patients, the isolate representing BSI was genetically related to the isolate representing VRE colonization: 12 pairs of isolates (52%) had identical banding patterns, 5 had closely related patterns, and 2 had possibly related patterns. CONCLUSION: Of the 768 patients colonized with VRE, 31 (4.0%) usually developed VRE BSI due to a related strain. Independent risk factors for BSI among colonized patients were admission from a long-term care facility, infection of an additional body site, and exposure to vancomycin. Independent risk factors for death were immunosuppression and VRE BSI. PMID- 18419362 TI - Cross-transmission of multidrug-resistant Acinetobacter baumannii clonal strains causing episodes of sepsis in a trauma intensive care unit. AB - OBJECTIVE: To investigate the mode of transmission of imipenem-resistant Acinetobacter baumannii strains causing episodes of sepsis. SETTING: A 7-bed trauma intensive care unit (ICU) in an orthopedic hospital in Greece. DESIGN: During a 14-week period (from January 10 to April 16, 2006), clinical specimens, along with samples taken on a weekly basis from the ICU environment and from the hands of health care workers (HCWs), were prospectively tested for imipenem resistant A. baumannii. Pulsed-field gel electrophoresis was used to study the genetic relatedness of the isolates recovered from these specimens and samples. RESULTS: During the survey, imipenem-resistant A. baumannii was identified in 14 hospitalized patients, from whom 40 multidrug-resistant and imipenem-resistant A. baumanii isolates were recovered. These pathogens caused episodes of bacteremia and sepsis in all but one of the patients and contributed to the death of 3 patients. Samples for culture were obtained from the environment and from the hands of HCWs; 29 imipenem-resistant A. baumannii isolates were recovered from the environment, and 12 from HCWs. One predominant genotype and 2 less predominant genotypes were detected among the 81 imipenem-resistant A. baumannii isolates. All 3 of these genotypes were found among patients and HCWs and were recovered from environmental samples. INTERVENTIONS: Control measures consisted of the closure of the ICU and the transfer of the patients to other units. The ICU was disinfected, and adherence to proper hand hygiene protocol was reinforced. These same clonal isolates were not recovered from clinical or environmental samples during the month after the reopening of the ICU. CONCLUSIONS: The extensive dissemination of imipenem-resistant A. baumannii clonal strains causing episodes of bacteremia and/or sepsis resulted from modes of transmission via multiple contaminated surfaces and objects and transiently colonized HCWs' hands. Closure of the ICU and its meticulous environmental decontamination led to the successful control of the outbreak. PMID- 18419363 TI - Outbreak of multidrug-resistant Serratia marcescens infection in a neonatal intensive care unit. AB - BACKGROUND: Serratia marcescens causes healthcare-associated infections and significant morbidity and mortality in neonatal intensive care units (NICUs). We report the investigation and control of an outbreak of multidrug-resistant (MDR) S. marcescens infection at an NICU. METHODS: An outbreak investigation and a case control study were undertaken at a 36-bed NICU in a tertiary care hospital in Baltimore, Maryland, for the period from October 2004 through February 2005. The outbreak investigation included case identification, review of medical records, environmental cultures, patient surveillance cultures, personnel hand cultures, and pulsed-field gel electrophoresis (PFGE). The case-control study included case identification and review of medical records. Infection control measures were implemented. Eighteen NICU neonates had cultures that grew MDR S. marcescens during the study period. The case-control study included 16 patients with the outbreak strain or an unidentified strain of MDR S. marcescens and 32 control patients not infected and/or colonized with MDR S. marcescens, treated in the NICU for at least 48 hours during the study period. RESULTS: PFGE analysis identified a single strain of MDR S. marcescens that infected or colonized 15 patients. Two patients had unique strains, and 1 patient's isolate could not be subtyped. An unrelated MDR S. marcescens isolate was recovered from a sink drain. Exposure to inhalational therapy was an independent risk factor for MDR S. marcescens acquisition after adjusting for birth weight. Extensive investigation failed to reveal a point source for the outbreak. CONCLUSION: A single epidemic strain of MDR S. marcescens spread rapidly and threatened to become endemic in this NICU. Transient carriage on the hands of healthcare personnel or on respiratory care equipment was the likely mode of transmission. Cohorting patients and staff, at the cost of bed closures and additional personnel, interrupted transmission and halted the outbreak. PMID- 18419364 TI - Infection control experience in a cooperative care center for transplant patients. AB - OBJECTIVE: To characterize infection control experience during a 6.5-year period in a cooperative care center for transplant patients. DESIGN: Descriptive analysis. SETTING: A cooperative care center for transplanted patients, in which patients and care partners are housed in a homelike environment, and care partners assume responsibility for patient care duties. PATIENTS: Nine hundred ninety one transplant patients. METHODS: Infection control definitions from the Centers for Disease Control and Prevention were used to ascertain infection rates. Environmental cultures were used to detect methicillin-resistant Staphylococcus aureus (MRSA), vancomycin-resistant Enterococcus (VRE), Clostridium difficile, and fungi during the first 18 months. Surveillance cultures were performed for a subset of patients and care partners. RESULTS: From June 1999 through December 2005, there were 19,365 patient-days observed. The most common healthcare-associated infection encountered was intravascular catheter-related bloodstream infection, with infection rates of 5.74 and 4.94 cases per 1,000 patient-days for hematopoietic stem cell transplant (HSCT) and solid organ transplant (SOT) patients, respectively. C. difficile-associated diarrhea was observed more frequently in HSCT patients than in SOT patients (3.97 vs 0.57 cases per 1,000 patient-days; P < .0001). There was no evidence of environmental contamination with MRSA, VRE, or C. difficile. Acquisition of MRSA was not observed. Acquisition of VRE was documented. CONCLUSION: This study documented that cooperative care was associated with some risk of healthcare associated infection, most notably intravascular catheter-associated bloodstream infection and C. difficile-associated diarrhea, it appears the incidences of these infections were roughly commensurate with those in other care settings. PMID- 18419365 TI - Lethal activity of nonthermal plasma sterilization against microorganisms. AB - OBJECTIVE: To determine the range and the mode of germicidal activity of sterilants generated by a nonthermal plasma sterilization system for microorganisms. METHODS: Representative bacteria, spores, viruses, bacteriophages, and fungi were exposed to the plasma cycle and the residual viability was measured in vitro. To assess the mode of lethal injury, Escherichia coli, Staphylococcus aureus, Bacillus atrophaeus, and bacteriophages were exposed to the plasma cycle, and the effects of the plasma-generated sterilants on the biological parameters were determined. RESULTS: There were at least 4-6 log reductions in viability for all microorganisms after 10 minutes of exposure to the plasma cycle. Electron micrographs and studies of the inhibition of bacteriophage infectivity suggested that the primary injury is to the organisms' cell envelopes. The plasma cycle also denatured isolated bacterial proteins and inactivated bacteriophages, but it had no effect on isolated DNA and bacterial proteins within exposed bacteria. CONCLUSION: Nonthermal plasma, which is produced at atmospheric temperature and pressure, generates sterilants that kill high concentrations of microorganisms and inactivate viruses during a 10-minute exposure. The primary injury appears to be at the surface structures of the organisms. This suggests that nonthermal plasma has utility for sterilization of heat-sensitive medical materials and devices. PMID- 18419366 TI - Reprocessing and reuse of single-use medical devices: a national survey of Canadian acute-care hospitals. AB - A national survey investigated the current practices of reprocessing and reusing single-use medical devices in Canadian acute-care hospitals. Our response rate was 72% (413 responses), and 28% of hospitals reprocess single-use devices. The results showed that Canadian hospital practices have not changed much in the past decade. PMID- 18419367 TI - University HealthSystem Consortium quality performance benchmarking study of the insertion and care of central venous catheters. AB - We report data from an observational benchmarking study of adherence to recommended practices for insertion and maintenance of central venous catheters at a heterogeneous group of academic medical centers. These centers demonstrated a need for significant improvement in implementation and documentation of quality performance measures for the prevention of catheter-related bloodstream infections. PMID- 18419368 TI - Some observations on the migration of Proteus mirabilis and other urinary tract pathogens over foley catheters. AB - The ability of uropathogens to migrate along external surfaces of silicone Foley catheters was examined. Proteus mirabilis and Pseudomonas aeruginosa were the most motile organisms. P. aeruginosa migrated over both triclosan-impregnated and nitrofurazone-impregnated catheters, but these antibacterials inhibited the migration of P. mirabilis. PMID- 18419369 TI - Interobserver variability in the assessment of clinical criteria for suspected urinary tract infection in nursing home residents. AB - We determined the interobserver variability in the assessment of clinical criteria for urinary tract infection (UTI) in nursing home residents. Pairs of nursing home staff caring for 30 residents were interviewed at the time UTI was suspected. At least one measure from each of 7 clinical criteria categories was reliably assessed by nursing home staff members. PMID- 18419370 TI - Methicillin-resistant Staphylococcus aureus infection in a college football team: risk factors outside the locker room and playing field. AB - We investigated a cluster of methicillin-resistant Staphylococcus aureus infections in college football players. Risk factors included a history of recurrent skin infections and contact with the skin lesions of persons outside college. The infections were controlled through treatment of carriers with topical mupirocin, chlorhexidine body washes, and enhancement of personal hygiene practices. Varsity and professional teams need to consider similar preventive measures. PMID- 18419371 TI - Incidence of methicillin-resistant Staphylococcus aureus nosocomial infections in intensive care units in Lyon University hospitals, France, 2003-2006. AB - We describe the trends in the incidence of methicillin-resistant Staphylococcus aureus nosocomial infection in intensive care units in Lyon hospitals from January 1, 2003, through December 31, 2006. The incidence rate decreased from 1.77 cases per 100 ICU patients in 2003 to 1.16 cases per 100 ICU patients in 2006, a reduction of 38.0% (P = .05). PMID- 18419372 TI - Improvement in prescribing habits and economic outcomes associated with the introduction of a standardized approach for surgical antimicrobial prophylaxis. AB - A quasi-experimental before-after study was conducted to determine the impact on prescribing habits and economic outcomes of an intervention that required use of an order form for surgical antimicrobial prophylaxis. Implementation of this intervention improved compliance with guidelines for prevention of surgical site infections, particularly with respect to choice of antibiotic, duration of use, and dosage, and resulted in decreased cost of surgical prophylaxis. PMID- 18419373 TI - Effect of building construction on Aspergillus concentrations in a hospital. AB - Air samples taken in a hospital undergoing construction and analyzed with a quantitative polymerase chain reaction (qPCR) assay for the Aspergillus genus did not show elevated concentrations of Aspergillus or particulate matter with a diameter of 5 microm or less in patient areas. Air samples from the construction zone indicated the containment system, which used polyethylene film barrier and negative pressure, was effective. PMID- 18419374 TI - Effect of accessibility of influenza vaccination on the rate of childcare staff vaccination. AB - We assessed the impact of free on-site influenza vaccination on childcare staff vaccination prevalence using 2 before-and-after studies. Vaccination was offered during the 2003-2004 and 2006-2007 influenza seasons. Staff vaccination prevalence was higher in each intervention season compared to the prior, nonintervention season. No baseline characteristics were associated with receipt of vaccination. PMID- 18419375 TI - Should infection control teams enforce an antimicrobial stewardship program for all clinicians? PMID- 18419376 TI - Simple quality improvement interventions reduce unnecessary intravascular device dwell time. PMID- 18419377 TI - Legionella colonization of the respiratory tract in patients without nosocomial exposure. PMID- 18419379 TI - Disinfectants containing chlorine: an occupational hazard? PMID- 18419378 TI - Impact of knowledge and positive attitudes about avian influenza (H5N1 virus infection) on infection control and influenza vaccination practices of Thai healthcare workers. PMID- 18419380 TI - Prevalence of nosocomial infections in adult intensive care units at a Kosova teaching hospital. PMID- 18419381 TI - Challenges to implementing second-dose varicella vaccination during an outbreak in the absence of a routine 2-dose vaccination requirement--Maine, 2006. AB - In June 2005, the Advisory Committee on Immunization Practices (ACIP) recommended administering a second dose of varicella vaccine during outbreaks, supplementing the routine 1-dose requirement. From October 2005 to January 2006, a varicella outbreak occurred in Maine in a highly vaccinated elementary school population. We investigated the outbreak, held a school-based vaccination clinic, and assessed costs in implementing ACIP's outbreak-response recommendation. Parents completed questionnaires and case investigation interviews. Personnel at the Maine Center for Disease Control and Prevention, the school in which the outbreak occurred ("school A"), and physician offices completed economic surveys. Forty eight cases occurred, with no hospitalizations or deaths. Vaccine effectiveness was 86.6% (95% confidence interval, 82.0%-90.1%). Of 240 eligible students, 132 (55.0%) received second-dose vaccination. Implementing ACIP's outbreak-response recommendation was challenging and cost approximately $26,875. Additionally, the routine 1-dose varicella vaccination policy did not confer adequate population immunity to prevent this outbreak. These findings support ACIP's June 2007 recommendation for a routine 2-dose varicella vaccination program. PMID- 18419382 TI - Varicella outbreak reporting, response, management, and national surveillance. AB - Two national surveys were conducted to evaluate the status of varicella case based surveillance and outbreak response. Although progress toward national surveillance has been significant, a large number of jurisdictions are still without case-based surveillance. For jurisdictions beginning case-based surveillance with limited resources, a staged approach is recommended. The national outbreak survey showed that a significant number of varicella outbreaks continue to occur. The majority of jurisdictions respond to these outbreaks, although the response varies considerably. Depending on the outbreak-response approach, costs per outbreak ranged from $3000 for a typical, or passive, response to $6000 for a more active response. As varicella surveillance and outbreak control improves, jurisdictions may benefit from more-standardized outbreak-control practices. The recent recommendation by the Advisory Committee on Immunization Practices for a routine second dose of varicella vaccine should lead to better varicella disease control, making case-based surveillance and appropriate outbreak response even more feasible. PMID- 18419383 TI - Varicella outbreak epidemiology in an active surveillance site, 1995-2005. AB - We describe trends and characteristics of varicella outbreaks identified in an active surveillance site from 1995 to 2005. Cases of varicella were reported to the active surveillance project, and outbreaks were defined retrospectively as > or =5 varicella cases epidemiologically linked to a common setting that occurred within 1 incubation period. Outbreaks were grouped by calendar year. From 1995 1998 to 2002-2005, varicella outbreaks significantly decreased in number, from 236 to 46 (P< .001); in size, from a median of 15 cases/outbreak to 9 cases/outbreak (P< .001); and in duration, from 44.5 days to 30 days (P< .001). The median age of case patients with outbreak-related varicella increased from 6 to 9 years (P< .001). The 1-dose varicella vaccination program has been successful in decreasing the number of outbreaks and cases; however, challenges remain with regard to controlling outbreaks among vaccinated persons and targeting vaccination efforts to susceptible persons in older age groups. PMID- 18419384 TI - Epidemiology of varicella hospitalizations in the United States, 1995-2005. AB - To describe the impact of the varicella vaccination program on varicella-related hospitalizations (VRHs) in the United States, data from the Varicella Active Surveillance Project (VASP) were used to compare rates of hospitalization and rates of complications among patients hospitalized for varicella-related conditions from 1995 to 2005. Of the 26,290 varicella cases reported between 1995 and 2005, 170 cases resulted in VRHs, including 1 case that resulted in death. Both VRH rates per 100,000 population and complications during VRH per 100,000 population decreased significantly between the early vaccination period (1995 1998) and the middle/late vaccination period (1999-2005). Infants and adults were at highest risk for VRH, and having been vaccinated against varicella was a protective factor. Varicella vaccination may have prevented a significant number of VRHs. The fact that 4 vaccinated children required hospitalization for varicella-related complications demonstrates that 1 dose of varicella vaccine does not prevent serious disease in all cases, even among previously healthy children. PMID- 18419385 TI - Varicella disease among vaccinated persons: clinical and epidemiological characteristics, 1997-2005. AB - Approximately 1 in every 5 children who receives 1 dose of varicella vaccine may develop varicella disease, also known as breakthrough disease, if exposed to varicella-zoster virus. Currently, in communities with high vaccination coverage, varicella cases mostly occur in vaccinated individuals. We report on the first population-based description of the clinical and epidemiological characteristics of varicella in populations with increasing vaccine coverage between 1997 and 2005. In vaccinated children 1-14 years of age, varicella was most often mild and modified; the atypical disease presentation may result in diagnostic challenges to health care providers. However, despite the generally mild nature of these cases, approximately 25% caused >50 lesions, and some resulted in serious complications similar to those occurring in unvaccinated individuals. Continued surveillance of the risk and characteristics of breakthrough disease will be needed, to monitor the effect of the new 2-dose vaccine recommendation for children. PMID- 18419386 TI - Laboratory diagnosis and characteristics of breakthrough varicella in children. AB - The atypical features of varicella in vaccinated persons (breakthrough varicella [BTV]) present diagnostic challenges. We examined varicella-zoster virus (VZV) polymerase chain reaction (PCR) and immunoglobulin (Ig) M and IgG serologic test results for confirming BTV cases. Among 33 vaccinated children with varicella like rash, we identified wild-type VZV in 58% overall and in 76% of those with adequate tissue specimens; no vaccine-type virus was found. Of the 12 subjects with PCR-confirmed BTV and acute-phase serum samples, 9 had detectable IgM, and all had highly elevated acute-phase IgG titers. Six subjects with negative PCR results had lower IgG titers and negative IgM results. Although PCR is the preferred method for laboratory confirmation of BTV, a positive serum varicella IgM test result should also be considered to be diagnostic in a suspected BTV case; however, a negative IgM test result cannot be used to rule out the diagnosis. The value of highly elevated IgG titers needs further evaluation. Larger studies are needed to confirm these results. PMID- 18419387 TI - Lack of association between age at varicella vaccination and risk of breakthrough varicella, within the Northern California Kaiser Permanente Medical Care Program. AB - BACKGROUND: Varicella vaccine currently is recommended for children between 12 and 18 months of age. However, rates of breakthrough varicella have been reported to be higher among children vaccinated before 14 or 15 months of age and to increase with time since vaccination. METHODS: An ongoing study at the Northern California Kaiser Permanente Medical Care Program is evaluating vaccine efficacy in 7585 children vaccinated with Varivax in 1995, when they were between 12 and 23 months of age. Cases of chickenpox are identified by telephone interviews with each child's parent(s) every 6 months. Mean age at varicella onset and mean time from vaccination to onset were calculated on the basis of age, in months, at vaccination. Logistic regression was used to test for trend, and the chi2 test was used to test for differences in rates of breakthrough varicella by age. RESULTS: Over the first 8 years of the study, a total of 1161 cases of breakthrough varicella were reported, for an average rate of 21.7 cases/1000 person-years. Vaccine effectiveness was 83.6% at year 8. The rate of breakthrough varicella did not change for each additional month of age at vaccination (P = .864), and no difference in the rate of breakthrough varicella was found between children vaccinated at <15 months of age and those vaccinated at > or =15 months of age. CONCLUSIONS: Our data do not show a difference in vaccine effectiveness with age at vaccination and thus support the current recommendations for initial vaccination between 12 and 18 months of age. PMID- 18419388 TI - Humoral and cell-mediated immune responses in children and adults after 1 and 2 doses of varicella vaccine. AB - Humoral and cell-mediated immune responses to varicella-zoster virus (VZV) have been evaluated after 1 and 2 doses of live attenuated varicella vaccine, Oka strain, in several studies. One dose of varicella vaccine, however, elicits detectable immune responses that are low and, in some cases, may be insufficient for complete protection against the virus after the normal decline in humoral and cell-mediated immunity with time. In contrast, immune responses after 2 doses are significantly higher and approximate the levels seen after natural disease. These investigations of vaccine-induced immunity suggest that 2 doses of VZV vaccine will better achieve the goals of the VZV vaccination program, by reducing the VZV burden of disease in childhood and preventing accumulation of young adults who are susceptible to or only partially protected from varicella. PMID- 18419389 TI - Use and limitations of varicella-zoster virus-specific serological testing to evaluate breakthrough disease in vaccinees and to screen for susceptibility to varicella. AB - A plethora of tests for determining the presence of antibodies to varicella zoster virus (VZV) have been developed over the years, with a wide range of performance standards. There is general agreement that the presence of VZV antibodies in serum indicates immunity to varicella and protection from chickenpox, although the role of specific antibody in mediating protection remains unclear. Both antibodies and cellular immunity probably interact to mediate immunity to the virus. In any case, VZV-specific serum antibody is a useful indicator of protection against chickenpox in patients and persons at high risk of exposure, whether they have been immunized or naturally infected. Serological tests are also a useful implement for evaluating the length of time that immunity to varicella persists after vaccination and whether waning of vaccine-induced immunity occurs. The purpose of this review is to contrast the strengths and weaknesses of currently available VZV antibody assays. Although several of these methods are useful for various specific applications, simpler and more accurate tests to measure antibodies to VZV are a high priority for future research and development. PMID- 18419390 TI - Varicella immunogenicity with 1- and 2-dose regimens of measles-mumps-rubella varicella vaccine. AB - A quadrivalent vaccine combining measles, mumps, rubella, and varicella antigens (MMRV) was developed to increase the coverage of varicella vaccine and reduce the number of injections children receive. Although the varicella antigen is as immunogenic in the latest formulation of MMRV vaccine as when it is administered alone, up to 14% of vaccine recipients do not achieve protective levels of anti varicella antibodies after a single dose, which can result in breakthrough varicella. A second dose of varicella vaccine raises response rates to 99% and was recently recommended by the Advisory Committee on Immunization Practices. Giving the second dose 3 months after the first (at approximately 15 months of age) would provide more protection against varicella but would necessitate a change in the childhood vaccination schedule, which currently calls for a second dose of MMRV vaccine between the ages of 4 and 6 years. PMID- 18419391 TI - An economic analysis of the universal varicella vaccination program in the United States. AB - Frequent varicella outbreaks with sizable impact on the US public health system have continued to occur despite the success of the country's 1-dose varicella vaccination program. The Advisory Committee on Immunization Practices recently recommended adding a routine second dose of varicella vaccine and weighed economic projections as well as public health goals in their deliberations. This decision-tree-based analysis was conducted to evaluate the economic impact of the projected 2-dose varicella vaccination program as well as the existing 1-dose program. The analysis used population-based vaccination coverage and disease incidence data to make projections for a hypothetical US birth cohort of 4,100,000 infants born in 2006. Compared with no vaccination, both the 1-dose program (societal benefit-cost ratio [BCR], 4.37) and 2-dose program (BCR, 2.73) were estimated to be cost saving from the societal perspective. Compared with the 1-dose program, the incremental second dose was not cost saving (societal incremental BCR, 0.56). The incremental cost-effectiveness ratio for the second dose was $343 per case prevented, or approximately $109,000 per quality-adjusted life-year saved, and these results were sensitive to assumptions about vaccine effectiveness and prices. PMID- 18419392 TI - The safety profile of varicella vaccine: a 10-year review. AB - Varivax (varicella virus vaccine live [Oka/Merck]; Merck), a live attenuated varicella vaccine, is indicated for vaccination against varicella in appropriate individuals > or =12 months of age. The 10-year safety profile for Varivax is described using data submitted to Merck from routine global postmarketing surveillance, combined with information from a Varicella Zoster Virus Identification Program, which uses polymerase chain reaction (PCR) analysis to identify the presence and strain of VZV in selected specimens. There were 16,683 reports worldwide voluntarily submitted to Merck, for an overall reporting rate of 3.4 reports/10,000 doses of vaccine distributed. PCR analysis of vesicular rashes that occurred within the first 2 weeks after vaccination was more likely to identify wild-type varicella-zoster virus (VZV), whereas the presence of Oka VZV was generally associated with vesicular rashes that occurred 15-42 days after vaccination. Reports of breakthrough varicella that occurred >42 days after vaccination were associated with wild-type VZV. Among 697 herpes zoster reports, PCR analysis identified Oka VZV in 57 reports and wild-type VZV in 38 reports. There were no primary neurologic adverse events associated with Oka VZV. Secondary transmission of Oka VZV from vaccine recipients with postvaccination vesicular rashes was identified in 3 susceptible household contacts. Disseminated Oka VZV was identified in 6 immunocompromised patients and 1 patient with Down syndrome. This review has shown that the vaccine is generally safe and well tolerated. PMID- 18419393 TI - Safety of varicella vaccine after licensure in the United States: experience from reports to the vaccine adverse event reporting system, 1995-2005. AB - Widespread use of varicella vaccine in the United States could enable detection of rare adverse events not identified previously. We reviewed data from 1995 to 2005 from the Vaccine Adverse Event Reporting System, including data from laboratory analyses, to distinguish adverse events associated with wild-type varicella-zoster virus (VZV) versus those associated with vaccine strain. Almost 48 million doses of varicella vaccine were distributed between 1995 and 2005. There were 25,306 adverse events reported (52.7/100,000 doses distributed); 5.0% were classified as serious (2.6/100,000 doses distributed). Adverse events associated with evidence of vaccine-strain VZV included meningitis in patients with concurrent herpes zoster. Patients with genetic predispositions may rarely have disease triggered by receipt of varicella vaccine. Overall, serious adverse events reported after varicella vaccination continue to be rare and must be considered relative to the substantial benefits of varicella vaccination. Ongoing safety surveillance and further studies may shed light on some of the hypothesized associations. PMID- 18419394 TI - Varicella vaccine exposure during pregnancy: data from 10 Years of the pregnancy registry. AB - BACKGROUND: The Pregnancy Registry for Varivax (Merck) was established to monitor for congenital varicella syndrome or other birth defects in the offspring of women who were exposed to varicella vaccine while pregnant. METHODS: The registry receives voluntary reports from health care providers or consumers about women given the vaccine 3 months before or during pregnancy. Follow-up is conducted to obtain and classify pregnancy outcomes. All reports are evaluated for the presence of birth defects. Outcomes from prospectively reported pregnancy exposures are used to calculate rates and 95% confidence intervals. RESULTS: From 17 March 1995 through 16 March 2005, 981 women were enrolled. Pregnancy outcomes were available for 629 prospectively enrolled women. Among the 131 live births to varicella-zoster virus-seronegative women, there was no evidence of congenital varicella syndrome (rate, 0% [95% confidence interval [CI], 0%-6.7%]), and major birth defects were observed in 3 infants (rate, 3.7% [95% CI, 0.8%-10.7%]). CONCLUSIONS: Although the numbers of exposures are not sufficient to rule out a very low risk, data collected in the pregnancy registry to date do not support a relationship between the occurrence of congenital varicella syndrome or other birth defects and varicella vaccine exposure during pregnancy. PMID- 18419395 TI - Varicella vaccination in Japan, South Korea, and Europe. AB - The most extensive use of varicella vaccine has been in the United States and Canada, where it is universally recommended. However, a number of other countries now have recommendations for use of the vaccine, which has been expanding in Europe and Latin America. In this article, we review information concerning varicella vaccination in Japan, where the vaccine was first developed, and in South Korea and parts of Europe. Despite the worldwide availability of an efficient vaccine, varicella vaccination policy is highly variable from country to country. The recent development of a tetravalent vaccine against measles, mumps, rubella, and varicella could modify this variability in the future. It is evident that efforts to control varicella will spread gradually to all continents. PMID- 18419396 TI - Varicella vaccination in Australia and New Zealand. AB - Varicella-zoster virus has been responsible for a significant disease burden, including hospitalizations and deaths in Australia and New Zealand. Varicella vaccine has been available in Australia since 1999 and, since November 2005, has been funded under the National Immunisation Program for use in all children as a single dose at 18 months of age and in a school-based catch-up program at 10-13 years of age. Recent hospitalization data from Australia show a decline in varicella hospitalizations in children 1-4 years of age, most likely related to vaccination. In New Zealand, varicella vaccine has been available since 1999 but is currently not recommended or funded on the New Zealand national immunization schedule. The anticipated licensure of combination measles-mumps-rubella varicella vaccines in both countries may lead to future schedule changes. PMID- 18419397 TI - Risk of herpes zoster in adults immunized with varicella vaccine. AB - A program of routine varicella vaccination of children 12-18 months of age, begun in the United States in 1995, has been very successful in reducing the incidence of varicella. Varicella-zoster virus (VZV), in both wild-type and live attenuated forms, is notable for its ability to produce latent infection of sensory neurons from which it can later reactivate to cause herpes zoster (HZ). Therefore, the effects of vaccination on this secondary VZV-related disease are important to consider; in practice, however, such studies are complicated by the typically long delay between acquisition of the virus and its reactivation. Studies of immunocompromised children have shown that vaccination is relatively protective against HZ in this highly vulnerable group. We now present long-term follow-up data on a group of individuals who received varicella vaccine as healthy young adults 10-26 years ago and who have been followed prospectively by means of active surveillance. Among some 2000 person-years of follow-up, 2 cases of HZ have occurred, for a rate of 1.00 case/1000 person-years. Overall, the incidence of HZ in this cohort, therefore, is similar to published data for the US population in the prevaccine era. PMID- 18419398 TI - Varicella vaccination of immunocompromised children. AB - BACKGROUND: Exposure of immunocompromised children to varicella often requires postexposure prophylaxis. Exposures requiring this management are often not recognized. Varicella can be a severe disease when it occurs in immunocompromised children, in spite of antiviral therapy. Varicella exposure and varicella in these children can also disrupt scheduled therapy for their underlying illness. Both postexposure prophylaxis and treatment of varicella are likely to be expensive and use significant medical resources. Numerous trials have been undertaken to vaccinate children who are immunocompromised by a variety of conditions and therapies that depress their immune function. METHODS: Clinical trials of varicella vaccine administration to immunocompromised children that were reported since 1975 were identified in the Ovid medical database. Reports were selected for analysis and discussion on the basis of their completeness and the utility of their conclusions. RESULTS: Vaccination before immune compromise is discussed as a strategy for some settings. The obstacles, potential opportunities, and success in varicella vaccination for immunocompromised children are separately analyzed for (1) children with leukemia and other malignancies, (2) human immunodeficiency virus-infected children, and (3) children with hematopoietic stem cell or solid-organ transplantation. CONCLUSIONS: Vaccination before immune compromise is often successful, and the vaccine-induced response is usually partially or fully protective. In many treatment settings, it is possible to safely vaccinate once the level of immune suppression has been reduced. Targets for future research are outlined. A consensus conference should be undertaken to develop guidelines for the use of varicella vaccine in immunocompromised children. PMID- 18419399 TI - The epidemiological, clinical, and pathological rationale for the herpes zoster vaccine. AB - Worldwide, herpes zoster (HZ) affects millions of patients (particularly older adults) annually and causes significant suffering due to acute and chronic pain, or postherpetic neuralgia (PHN). The objective of this article is to explain the rationale for the HZ vaccine by summarizing data on the epidemiology of HZ in the immunocompetent host, with a focus on recent incidence and risk factor studies; to review information on the burden of HZ; and to discuss the challenges of lessening the morbidity of the disease. The incidence and severity of HZ and PHN are highest in older adults. Given the central nervous system damage caused by HZ, the difficulty of adequately treating HZ to prevent PHN, and the intractability of PHN, the advent of the HZ vaccine appears to be a crucial innovation for preventing HZ and PHN. PMID- 18419400 TI - National survey of primary care physicians regarding herpes zoster and the herpes zoster vaccine. AB - BACKGROUND: This study describes physicians' perception of burden associated with herpes zoster (HZ) and postherpetic neuralgia (PHN), intentions for recommending the HZ vaccine, and perceived barriers to vaccination. METHODS: A national survey of 438 general internal medicine (GIM) and 433 family medicine (FM) physicians was conducted during November-December 2005. RESULTS: The survey response rate was 69%. Approximately 35% of GIM and FM physicians strongly agreed that HZ and PHN caused a significant burden of disease. For patients 60-79 years of age, > or =80% of GIM and FM physicians were somewhat or very likely to recommend HZ vaccine. In multivariate analyses, physicians who strongly agreed that HZ and PHN cause significant burden were more likely to recommend the vaccine to patients 60 79 years of age (odds ratio [OR], 2.75 [95% confidence interval [CI], 1.85 4.09]), whereas those who felt there was insufficient information about duration of protection (OR, 0.40 [CI, 0.24-0.67]), that the need to store HZ vaccine in a freezer was a definite barrier (OR, 0.31 [CI, 0.13-0.75]), or that their patients would not pay for the vaccine if it was not covered by insurance (OR, 0.57 [CI, 0.38-0.86]) were less likely to recommend it. CONCLUSIONS: Primary care physicians perceived a high level of burden from HZ and PHN and generally favored the HZ vaccine. PMID- 18419401 TI - The impact of the varicella vaccination program on herpes zoster epidemiology in the United States: a review. AB - Speculation that a universal varicella vaccination program might lead to an increase in herpes zoster (HZ) incidence has been supported by modeling studies that assume that exposure to varicella boosts immunity and protects against reactivation of varicella-zoster virus (VZV) as HZ. Such studies predict an increase in HZ incidence until the adult population becomes predominantly composed of individuals with vaccine-induced immunity who do not harbor wild-type VZV. In the United States, a varicella vaccination program was implemented in 1995. Since then, studies monitoring HZ incidence have shown inconsistent findings: 2 studies have shown no increase in overall incidence, whereas 1 study has shown an increase. Studies from Canada and the United Kingdom have shown increasing rates of HZ incidence in the absence of a varicella vaccination program. Data suggest that heretofore unidentified risk factors for HZ also are changing over time. Further studies are needed to identify these factors, to isolate possible additional effects from a varicella vaccination program. Untangling the contribution of these different factors on HZ epidemiology will be challenging. PMID- 18419403 TI - Strategies for herpes zoster vaccination of immunocompromised patients. AB - A vaccine to prevent herpes zoster (HZ) in adults > or =60 years of age with healthy immune systems was recently approved by the US Food and Drug Administration. This vaccine is contraindicated in persons with certain immunodeficiency states or who are receiving immunosuppressive therapy. On the basis of studies of the varicella vaccine in healthy and immunosuppressed children and studies of HZ vaccine in healthy adults before its licensure, a series of strategies are proposed for evaluating the live HZ vaccine in immunosuppressed persons. In addition, the use of other vaccines, including heat inactivated or replication-defective varicella-zoster virus to prevent HZ in immunocompromised persons, is also discussed. PMID- 18419402 TI - Vaccination against Herpes Zoster and Postherpetic Neuralgia. AB - BACKGROUND: Herpes zoster (HZ) and postherpetic neuralgia (PHN) cause significant morbidity in older adults. The incidence and severity of HZ and PHN increase with age in association with an age-related decline in varicella-zoster virus (VZV) specific cell-mediated immunity (VZV-CMI). VZV vaccines can boost VZV-CMI. Therefore, we tested the hypothesis that VZV vaccination would protect older adults against HZ and PHN. METHODS: We enrolled 38,546 adults > or =60 years of age in a randomized, double-blind, placebo-controlled trial of an investigational HZ vaccine and actively followed subjects for the development of HZ. The primary end point was the burden of illness due to HZ (HZ BOI), a composite measure of the incidence, severity, and duration of pain and discomfort caused by HZ. The secondary end point was the incidence of PHN. RESULTS: Subject retention was >95%. HZ vaccine reduced the HZ BOI by 61.1% (95% confidence interval [CI], 51.1% 69.1%; P<.001) and reduced the incidence of PHN by 66.5% (95% CI, 47.5%-79.2%; P<.001). The incidence of HZ was also reduced by 51.3% (95% CI, 44.2%-57.6%; P<.001). HZ vaccine was well tolerated; injection site reactions were generally mild. HZ vaccine neither caused nor induced HZ. CONCLUSION: The Shingles Prevention Study demonstrated that HZ vaccine significantly reduced the morbidity due to HZ and PHN in older adults. PMID- 18419404 TI - Perspective on live varicella vaccine. AB - The attenuation of varicella-zoster virus (VZV) by Takahashi in 1974 was a remarkable achievement. It swiftly led to development of a live vaccine against chickenpox, which was initially tested in Japan. With its successful employment in immunocompromised children to prevent morbidity and mortality due to varicella, the vaccine began to be tested in healthy children in Japan and elsewhere. In the United States, vaccine use progressed from extensive clinical trials that demonstrated safety and efficacy to universal immunization of healthy infants and children. In the past 10 years, >30 million healthy American individuals, mostly children, have been vaccinated. With increasing use of vaccine, there has been a concomitant decrease in the incidence of disease, along with decreases in hospitalizations and deaths due to VZV. To improve protection, however, a 2-dose schedule of immunization was recommended for routine use in all children by the Centers for Disease Control and Prevention in June 2006. At roughly the same time, licensure of the combined measles-mumps-rubella-varicella vaccine was completed, which allowed harmonization of immunization against these 4 viruses with 1 injection given twice in childhood. Concomitantly, a version of the varicella vaccine with 10 times the titer was developed for immunization of healthy individuals >60 years of age against herpes zoster (HZ). Although elimination of VZV from human populations may not yet be possible, the combined approach of immunization against both varicella in childhood and HZ in adulthood in the developed world are predicted to dramatically increase our control of this troublesome virus. PMID- 18419405 TI - Varicella vaccine in the United States: a decade of prevention and the way forward. PMID- 18419406 TI - Development of varicella vaccine. AB - The Oka strain of varicella-zoster virus (VZV) was first isolated from vesicles of an otherwise healthy 3-year-old boy with typical varicella. The virus was passaged 11 times in human embryonic lung fibroblasts at 34 degrees C and 12 times in guinea pig embryo fibroblasts (GPEFs) at 37 degrees C. GPEFs were the only nonprimate cells tested in which some degree of viral replication occurred. The resultant virus was temperature sensitive and showed host dependency, measured as better replication in GPEFs than that shown by the parental virus. The passaged virus was used as a candidate varicella vaccine and proved safe and effective for healthy and immunocompromised children. During the follow-up of vaccinated children with acute lymphocytic leukemia, the incidence of herpes zoster (HZ) was significantly lower among children who did not have a rash after vaccination, compared with those who had a rash caused by VZV (6 [2.3%] of 260 vs. 12 [17.1%] of 70, respectively). Because of the pathogenesis of VZV, the incidence of latency and of HZ is predicted to be lower among vaccine recipients than among individuals who have experienced varicella. PMID- 18419407 TI - Molecular analysis of the Oka vaccine strain of varicella-zoster virus. AB - Live attenuated Oka vaccine was subjected to molecular analysis, with the long term goal of identifying genes in the vaccine strain responsible for its attenuation. Complete genomic sequences for both the Oka parent virus and the Oka vaccine virus were determined and compared. There were differences in only 42 bases between the 2 viruses. More than one-third (15/42) of the nucleotide substitutions in V-Oka were found in open reading frame (ORF) 62. These differences were also observed when sequences of Japanese low-passage clinical isolates of varicella-zoster virus (VZV) were compared with the Oka vaccine strain. The ORF62 gene encodes an immediate early (IE) protein, IE62, that is the major transactivator of VZV and, as such, is critical to initiating the VZV gene expression cascade. Several insertions and deletions were also observed in comparing the 2 sequences, largely in the internal tandem repeat units. Functional differences between the 2 types of Oka virus were also examined. Oka vaccine was found to consist of a mixture of different subpopulations, and the parental Oka virus replicates more efficiently than does the vaccine strain. Development of a bacterial artificial chromosome for VZV expression should lead to the discovery of additional differences in wild and vaccine types and, thus, enhance our understanding of the genetic basis for attenuation of the Oka vaccine strain. PMID- 18419408 TI - Comparative analyses of the 9 glycoprotein genes found in wild-type and vaccine strains of varicella-zoster virus. AB - The complete DNA sequences of wild-type and vaccine strains of varicella-zoster virus have been published and listed in GenBank. In this comparative genomic analysis, the sequences of the 9 glycoprotein open reading frames (ORFs) were compared. They included gE (ORF68), gI (ORF 67), gC (ORF14), gH (ORF37), gL (ORF60), gB (ORF31), gK (ORF5), gM (ORF50), and gN (ORF8 or ORF9A). After realignment on the basis of newer data, the corrected gB sequence was lengthened to include 931 residues. The data showed that there were glycoprotein polymorphisms that differentiated North American/European strains from Japanese strains-for example, an additional ATG codon in the gL of all Oka strains. Also, there were a small number of coding single-nucleotide polymorphisms present only in glycoproteins of vaccine strains. Because these changes were highly conserved, the structure of the glycoprotein was unlikely to be altered. PMID- 18419409 TI - Vaccine Oka variants and sequence variability in vaccine-related skin lesions. AB - As with most live attenuated viral vaccines, varicella vaccine comprises a mixture of variant strains. Knowledge about the pathogenic potential of individual strains in the varicella vaccine is limited. Vaccination against chickenpox causes a usually modified varicella-like rash in a small percentage of healthy children, and vaccine virus reactivates on rare occasions to cause herpes zoster (HZ). In several published studies, our respective laboratories have analyzed genomic variation among specimens from cases of postvaccination rash and HZ in vaccine recipients, focusing on polymorphisms between vaccine Oka strains and the parental Oka strain. In most respects, these studies were in close agreement, identifying the set of wild-type markers among vaccine adverse event isolates, each occurring at similar frequencies. The same 3 universally present vaccine markers, at positions 106262, 107252, and 108111, were also identified by both laboratories. One notable difference has been the observation of mostly clonal vaccine virus among isolates examined by one laboratory and mostly mixed viruses in isolates examined by the other. In addition to reviewing and comparing our combined observations, we propose possible explanations for our contrasting findings and propose future studies to reconcile them. PMID- 18419410 TI - Humoral and cellular immunity to varicella-zoster virus: an overview. PMID- 18419411 TI - A model of lytic, latent, and reactivating varicella-zoster virus infections in isolated enteric neurons. AB - Because human primary afferent neurons are not readily obtained, we sought to develop a model in which the lytic, latent, and reactivating phases of varicella zoster virus (VZV) infection were recapitulated in neurons from an animal source. Enteric neurons were obtained from the small intestine of adult guinea pigs and from the bowel of fetal mice. Latency was established when these neurons were infected by cell-free VZV in the absence of fibroblasts or other cells of mesodermal origin. In contrast, lytic infection ensued when fibroblasts were present or when the enteric neurons were infected by cell-associated VZV. Latency was associated with the expression of a limited subset of viral genes, the products of which were restricted to the cytoplasm. Lysis was associated with the expression of viral glycoproteins, nuclear translocation of latency-associated gene products, and rapid cell death. Reactivation was accomplished by expressing VZV open reading frame (ORF) 61p or herpes simplex virus ICP0 in latently infected neurons. Isolated enteric neurons from guinea pigs and mice recapitulate latent gene expression in human cranial nerve and dorsal root ganglia. Expression of latency-associated VZV gene products was detected in 88% of samples of adult human intestine, suggesting that VZV not only infects enteric neurons but also is latent in the human enteric nervous system. This in vitro model should facilitate further understanding of latency and reactivation of VZV. PMID- 18419412 TI - Knowledge, attitudes, and practices regarding varicella vaccination among health care providers participating in the varicella active surveillance project, Antelope Valley, California, 2005. AB - Knowledge, attitudes, and practices regarding varicella vaccination and disease were assessed among health care providers participating in the Varicella Active Surveillance Project in Antelope Valley, California, in 2005. Compared with those of a similar survey conducted in 1999, results suggest a reduction in concerns about vaccine safety and efficacy. Routine assessment of adolescents for varicella susceptibility was reported by 87% of respondents, but only 42% reported routine assessment of adults. Several respondents were unaware that disease in a vaccinated person is infectious, and some did not know the vaccination recommendations pertaining to susceptible health care workers, suggesting a need for provider education on these issues. PMID- 18419413 TI - Changing varicella epidemiology in active surveillance sites--United States, 1995 2005. AB - Significant reductions in varicella incidence were reported from 1995 to 2000 in the varicella active surveillance sites of Antelope Valley (AV), California, and West Philadelphia (WP), Pennsylvania. We examined incidence rates, median age, and vaccination status of case patients for 1995-2005. Coverage data were from the National Immunization Survey. By 2005, coverage among children 19-35 months of age reached 92% (AV) and 94% (WP); 57% and 64% of case patients in AV and WP, respectively, were vaccinated; and varicella incidence declined by 89.8% in AV and 90.4% in WP. Incidence declined in all age groups, especially among children <10 years of age in both sites and among adolescents 10-14 years of age in WP. In AV, since 2000, the incidence among adolescents 10-14 and 15-19 years of age increased. Implementation of school requirements through 10th grade in WP may explain the differences in the decline in incidence among adolescents. Continued surveillance will be important to monitor the impact that the 2-dose vaccine policy in children has on varicella epidemiology. PMID- 18419414 TI - Status of school entry requirements for varicella vaccination and vaccination coverage 11 years after implementation of the varicella vaccination program. AB - We reviewed progress toward adoption of day care and school entry requirements in each state and the District of Columbia (DC) and compared varicella vaccination coverage by state to year of implementation of day care entry requirements. By the start of the 2006-2007 school year, 46 states (92%) and DC had implemented entry requirements for varicella vaccination. Between 1997 and 2005, national varicella vaccination coverage among children 19-35 months of age increased from 25.8% to 87.9%. Implementation of day care entry requirements in 2000 or earlier was associated with higher vaccination coverage (> or =90%; P=.002). Implementation of day care and school entry requirements for varicella vaccination is an important strategy for achieving and maintaining high vaccination coverage among preschool- and school-aged children in the United States. The newly adopted vaccine policy recommendation of 2 doses of varicella vaccine for all school-aged children should be incorporated into the states' school entry requirements. PMID- 18419415 TI - Varicella vaccine effectiveness in the US vaccination program: a review. AB - Varicella vaccine (Varivax, Merck) has been available in the United States since 1995. We reviewed published results of postlicensure studies of vaccine effectiveness. Among 19 studies, 17 reported on the effectiveness of vaccine received before exposure, and 2 reported on effectiveness after exposure. Studies used retrospective and prospective cohort, case-control, and secondary attack rate (household contact) designs. The majority of estimates assessed protection against clinically diagnosed varicella. One dose of varicella vaccine was 84.5% effective (median; range, 44%-100%) in preventing all varicella and 100% effective (mean and median) in preventing severe varicella. When administered after exposure, varicella vaccine was highly effective in preventing or modifying varicella. Although 1 dose of varicella vaccine has provided excellent protection, a higher degree of effectiveness is needed in order to interrupt transmission and to prevent outbreaks in settings with high contact rates. Monitoring the effectiveness of the newly recommended 2-dose childhood vaccine schedule for varicella vaccine is a priority. PMID- 18419416 TI - Epidemiology of varicella in Connecticut, 2001-2005. AB - We analyzed varicella surveillance data in Connecticut for 2001-2005, to describe the epidemiology of varicella in a highly vaccinated population after the introduction of varicella vaccine and to determine the number of preventable cases that had occurred during school-related outbreaks. Overall, the incidence of varicella did not change during the surveillance period. Vaccination rates among reported case patients increased, and the severity of infection decreased. An annual median of 2.5 cases/outbreak was identified as being preventable, with a majority of these cases being preventable by revaccination of previously vaccinated persons. Continued surveillance is needed in order to monitor changing trends in varicella epidemiology. PMID- 18419417 TI - Varicella among adults: data from an active surveillance project, 1995-2005. AB - We report detailed population-based data on varicella among adults. In 2 US varicella active surveillance sites with high vaccine coverage among young children, the incidence of varicella among adults declined 74% during 1995-2005. A low proportion (3%) of adults with varicella had been vaccinated, with no improvement over the decade of program implementation, suggesting that the decline was likely secondary to herd-immunity effects. Compared with children, adults had more severe varicella in terms of both clinical presentation and frequency of complications. However, <30% of adults with varicella were treated with acyclovir. Among adolescents, illness severity was intermediate between that in children and adults. Varicella cases are preventable through vaccination. As we enter the second decade of the varicella vaccination program in the United States, we need to ensure that susceptible adolescents and adults are adequately protected from varicella by vaccination and that those who acquire varicella are appropriately treated with effective antiviral treatment. PMID- 18419420 TI - Who is steering the ship? External influences on infection control programs. AB - Over the past decade, hospital infection control programs have begun to face new external influences that affect day-to-day practice. The mainstream media's interest in hospital-acquired infection sparked consumer interest, which resulted in more legislative activity and government regulation. Industry's influence is also increasing. To meet the increased demands of external agencies, infection control programs will need additional resources, and the infection control community will need to be more proactive in educating the public and defining the priorities for practice and research. PMID- 18419421 TI - Mycobacterial and other serious infections in patients receiving anti-tumor necrosis factor and other newly approved biologic therapies: case finding through the Emerging Infections Network. AB - We present the results of a nationwide survey of infectious disease consultants to identify mycobacterial and other serious infections in patients receiving anti tumor necrosis factor compounds and other novel targeted therapies. Nontuberculous mycobacterial infections, histoplasmosis, and invasive Staphylococcus aureus infection were all reported more frequently than was tuberculosis disease in this context. PMID- 18419422 TI - Concordant HIV infection and visceral leishmaniasis in Ethiopia: the influence of antiretroviral treatment and other factors on outcome. AB - BACKGROUND: Coinfection with human immunodeficiency virus (HIV) and Leishmania donovani visceral leishmaniasis (VL) in Africa is an emerging, poorly understood disease. METHODS: We evaluated 356 consecutive patients coinfected with HIV and VL treated in Humera, northwest Ethiopia, from February 2003 to October 2006, for risk factors for VL relapse and death and the effect of antiretroviral therapy (ART). RESULTS: During 2928 patient-months of follow-up, 256 VL episodes and 39 deaths occurred. Among 195 patients receiving ART, 31.3% had > or = 1 VL episode, and 14.4% died. Among 161 patients who did not receive ART, 26.1% had > or = 1 VL episodes, and 6.8% died. A total of 54 patients who received ART and 58 patients who did not receive ART had > or = 1 VL relapse. VL relapse among patients receiving ART was associated with a baseline CD4 cell count < 100 cells/microL (hazard ratio [HR], 2.50; 95% confidence interval [CI], 1.21-5.15) and > or = 2 previous VL episodes (HR, 3.74; 95% CI, 1.40-10.02). Failure to clear parasites after VL treatment was usually followed by symptomatic VL relapse. Patients who relapsed showed poor CD4 cell count recovery while receiving ART. ART was partially protective against VL relapse (HR, 0.46; 95% CI, 0.26-0.82). However, 28% of first VL relapses while receiving ART occurred despite a CD4 cell count > 200 cells/microL; in 5% of VL relapses, the CD4 cell count had been > 200 cells/microL for > 6 months. Factors associated with all-cause mortality among patients receiving ART were baseline CD4 cell count < 100 cells/microL (HR, 3.20; 95% CI, 1.30-7.87) and VL episodes during follow-up (HR for 1 episode, 4.97 [95% CI, 2.09-11.86]; HR for > 2 episodes, 3.22 [95% CI, 1.01-10.23]). CONCLUSIONS: Concordant HIV infection and VL is a major, acquired immunodeficiency syndrome defining illness with high relapse and mortality rates; ART reduces relapses; and secondary antileishmanial prophylaxis may benefit patients at risk of relapse. PMID- 18419423 TI - Tumor necrosis factor-alpha at the intersection of mycobacterial immunity and pathogenesis: an important new address in medicine. PMID- 18419424 TI - Carbapenem-resistant Escherichia coli harboring Klebsiella pneumoniae carbapenemase beta-lactamases associated with long-term care facilities. AB - Nine carbapenem-resistant Escherichia coli isolates harboring Klebsiella pneumoniae carbapenemase (KPC)-2 or KPC-3 enzymes were identified in patients residing in 7 distinct long-term care facilities. Cefotaxime-hydrolyzing (CTX-M) type beta-lactamases were also documented in 3 isolates. The identification of these enzymes in patients staying in long-term care facilities should be of great concern to all components of health care systems. PMID- 18419427 TI - Reversed halo sign in invasive pulmonary fungal infections. AB - Computed tomography scans of documented pulmonary mold infections were reviewed for the presence of the reversed halo sign, a focus of ground-glass attenuation surrounded by a solid ring. The reversed halo sign was an early sign, seen in approximately 4% of patients with pulmonary mold infections, usually with zygomycosis. PMID- 18419428 TI - Preexisting psychological stress predicts acute and chronic fatigue and arthritis following symptomatic parvovirus B19 infection. AB - BACKGROUND: Psychological stress is thought to be an important factor in the pathogenesis of chronic fatigue syndrome/myalgic encephalomyelitis (CFS/ME). Therefore, we sought to examine this relationship in the context of parvovirus B19 infection. METHODS: Thirty-nine patients with laboratory-documented acute parvovirus B19 infection were asked to complete questionnaires on negative life events, perceived stress, and negative affect relevant to the time of onset of parvovirus infection and during the preceding 12 months. These scores were combined into an overall stress index, which was then examined for associations with particular parvovirus-associated symptoms at acute infection and during the ensuing 1-3 years. Additional characteristics monitored included presence of parvovirus antibodies and nucleic acid, cortisol level, dehydroepiandrosterone level, autoantibodies, levels of a range of serum cytokines, and human leukocyte antigen class I and II alleles. RESULTS: Stress index was significantly associated with development of fatigue during the acute phase of parvovirus B19 infection and also with chronic fatigue and arthritis occurring 1-3 years following acute parvovirus B19 infection. Logistic regression that included all clinical variables indicated that a high stress index at the time of onset of infection was the primary predictor of CFS/ME 1-3 years following acute parvovirus B19 infection (odds ratio, 25.7; 95% confidence interval, 1.7-121.9; P=.005). CONCLUSIONS: We report a highly significant association between psychological stress and development of acute and chronic fatigue and arthritis several years following laboratory-documented acute parvovirus B19 infection. PMID- 18419429 TI - Entecavir therapy induces de novo HIV reverse-transcriptase M184V mutation in an antiretroviral therapy-naive patient. AB - Recently, entecavir was introduced as a potent drug against hepatitis B virus infection. Initially, it was suggested not to have any effect on human immunodeficiency virus (HIV) infection. This guideline was revised in 2007 because of a report showing that the M184V mutation was selected in an hepatitis B virus and HIV-coinfected patient previously treated with lamivudine. Our investigation revealed findings similar to those preveiously reported but in an antiretroviral therapy-naive patient coinfected with HIV and hepatitis B virus. After 26 weeks of entecavir therapy, the M184V mutation dominated the plasma viral population. Thus, entecavir should only be used for coinfected patients who simultaneously receive suppressive therapy against HIV infection. PMID- 18419430 TI - Spontaneous splenic rupture caused by Babesia microti infection. AB - Babesiosis has not been previously associated with spontaneous splenic rupture. We describe an otherwise healthy 61-year-old man with symptomatic babesiosis whose spleen ruptured during hospitalization. Although this complication is rare, practitioners who commonly treat patients with babesiosis should be aware of its potential occurrence. PMID- 18419432 TI - Bat rabies in the United States and Canada from 1950 through 2007: human cases with and without bat contact. AB - BACKGROUND: Since the 1980s, rare cases of rabies in humans in Canada and the United States have been almost exclusively caused by the bat-variant virus. METHODS: We reviewed indigenously acquired cases of bat-variant rabies in humans in Canada and the United States from 1950 through 2007. RESULTS: Of 61 cases identified, 5 occurred after organ transplantation and were excluded from further analysis. A bite was reported by 22 (39%) of the case patients, 9 (16%) had a direct contact (i.e., were touched by a bat) but no history of a bite, 6 (11%) found bats in their home (2 [4%] in the room where they slept) but reported no direct contact, and 19 (34%) reported no history of bat exposure whatsoever. With the exception of California (8 cases) and Texas (7 cases), no state or province had >3 cases. Of the case patients, 76% were men, and 40% were 10-29 years of age. The median incubation period was 7 weeks (<10 weeks in 72% of cases). The incidence of bat-variant rabies cases increased from 2.2 per billion person-years in 1950-1989 to 6.7 per billion person-years in 1990-2007. Of 36 case patients with bat rabies described since 1990, 16 had no history of direct bat contact; 2 (13%) of the 16 would have qualified for rabies postexposure prophylaxis on the basis of exposure criteria expanded in 1995 to include bats that were in the same room as a sleeping person. The incidence of rabies for this type of exposure was 0.6 cases per billion person-years. CONCLUSION: The true preventable proportion of cases and the number needed to treat with rabies postexposure prophylaxis to prevent 1 case would be useful information to inform the current guidelines. PMID- 18419433 TI - Ocular manifestations associated with brucellosis: a 26-year experience in Peru. AB - BACKGROUND: Brucellosis has unusual clinical manifestations. Ocular involvement caused by brucellosis remains poorly recognized in areas in which brucellosis is endemic. METHODS: A prospective study was performed to evaluate patients attending the Instituto de Medicina Tropical "Alexander von Humboldt" and the Hospital Nacional Cayetano Heredia (Lima, Peru) from January 1980 through December 2005 who received a diagnosis of brucellosis with ocular involvement. Diagnosis was made on the basis of clinical findings as well as agglutinations and/or culture positive for Brucella melitensis. RESULTS: During a period of 26 years, 1551 patients with brucellosis were seen, including 52 patients with ocular brucellosis. We found that 7 (0.7%) of 965 patients with acute brucellosis and 45 (7.9%) of 570 patients with chronic brucellosis had ocular brucellosis (P<.001). In 16 patients with brucellosis, the disease stage was unclassified. The most frequent ocular presentation was uveitis, which was found in 43 (82.7%) of the 52 patients with ocular brucellosis. Posterior uveitis was the most frequent uveal syndrome (21 cases; 45.7%). Patients with panuveitis had the worst visual prognosis: 8 of 9 patients with panuveitis were legally blind, including 5 patients with no light perception. CONCLUSIONS: Brucellosis may involve the eye and can lead to serious complications. In patients with brucellosis, early ophthalmologic evaluation can lead to prompt treatment and might prevent blindness from severe ocular damage. PMID- 18419434 TI - Pneumococcal necrotizing pneumonia in Utah: does serotype matter? AB - BACKGROUND: Streptococcus pneumoniae is the most common cause of bacterial pneumonia in children. Despite the use of the 7-valent pneumococcal conjugate vaccine, the incidence of pneumococcal necrotizing pneumonia (PNP) has been increasing. Our objectives were to describe temporal trends in PNP and to evaluate pneumococcal serotypes associated with PNP in Utah. METHODS: We performed a retrospective review of all children <18 years of age who were cared for at a tertiary care children's hospital and who had blood, lung tissue, broncheoalveolar lavage, or pleural fluid cultures that grew S. pneumoniae, as well as radiographic evidence of pneumonia, from January 1997 through March 2006. All S. pneumoniae isolates were typed. RESULTS: A total of 124 children with pneumococcal pneumonia were identified, and 33 (27%) of these children had radiographic evidence of PNP. During the period 1997-2000, 5 (13%) of 39 cases of culture-confirmed pneumococcal pneumonia were associated with PNP. In contrast, during the period 2001-2006, 28 (33%) of 85 pneumococcal pneumonia cases were complicated by PNP (odds ratio, 3.34; 95% confidence interval, 1.11-12.03). Non-7 valent pneumococcal conjugate vaccine serotypes comprised 49% of the isolates during 1997-2000 and 88% of isolates during 2001-2006 (odds ratio, 7.89; 95% confidence interval, 2.91-21.90). Pneumonia due to serotype 3 was most often associated with PNP. Eleven (79%) of 14 cases of serotype 3-associated pneumonia were associated with PNP. When compared with all other serotypes, serotype 3 was strongly associated with necrosis (odds ratio, 14.67; 95% confidence interval, 3.39-86.25). CONCLUSIONS: PNP is a serious and increasingly common complication of S. pneumoniae pneumonia in Utah. Infection with serotype 3 is associated with an increased risk of developing PNP. The increase in the incidence of infection due to nonvaccine serotypes reported worldwide and the changing epidemiology of invasive pneumococcal disease should be considered when developing vaccine strategies. PMID- 18419435 TI - Replacement pneumococcal disease in perspective. PMID- 18419436 TI - Community-acquired Legionella pneumonia: new insights from the German competence network for community acquired pneumonia. AB - BACKGROUND: The Competence Network for Community Acquired Pneumonia (CAPNETZ) offers a unique opportunity to study the epidemiology of legionellosis throughout Germany, applying sophisticated diagnostic tools. METHODS: The incidence, clinical characteristics, and outcome of Legionella pneumonia in 2503 adult patients with community-acquired pneumonia, participating in the German Multicenter Study of the CAPNETZ, were studied. RESULTS: Legionella pneumonia was diagnosed in 94 patients (3.8%), thus identifying Legionella species as one of the most common pathogens to cause community-acquired pneumonia. It was equally common among ambulatory and hospitalized patients (3.7% and 3.8%, respectively). The predominant species causing community-acquired pneumonia was Legionella pneumophila; however, 10% of cases were caused by other species not detectable by the urinary antigen test. Patients whose disease was diagnosed by urinary antigen testing experienced a more severe clinical course. Compared with hospitalized patients, ambulatory patients with Legionella pneumonia showed an equal sex distribution, were younger, had fewer comorbidities, fewer cases of discordant initial antimicrobial treatment, and a milder clinical course without fatalities. Thirty percent of patients with Legionella pneumonia received discordant initial antimicrobial treatment without increased mortality. CONCLUSIONS: Legionella is a leading cause of community-acquired pneumonia in Germany. It needs to be considered equally in hospitalized and ambulatory patients. A positive result of a urine antigen test is associated with a more severe clinical course and leads to a potentially relevant underrecognition of species other than L. pneumophila. Legionella pneumonia in outpatients differs significantly from that in hospitalized patients in terms of clinical presentation and outcome. There was an unacceptably high rate of discordant initial antimicrobial treatment. PMID- 18419437 TI - Community-acquired legionnaires disease: implications for underdiagnosis and laboratory testing. PMID- 18419438 TI - Epidemiology of Staphylococcus aureus colonization in nursing home residents. AB - BACKGROUND: We sought to characterize the clinical and molecular epidemiologic characteristics of Staphylococcus aureus colonization (especially extranasal colonization) and to determine the extent to which community-associated methicillin-resistant S. aureus (MRSA) has emerged in community nursing homes. METHODS: The study enrolled a total of 213 residents, with or without an indwelling device, from 14 nursing homes in southeastern Michigan. Samples were obtained from the nares, oropharynx, groin, perianal area, wounds, and enteral feeding tube site. Standard microbiologic methods were used to identify methicillin-susceptible S. aureus and MRSA. Molecular epidemiologic methods included pulsed-field gel electrophoresis, PCR detection of Panton-Valentine leukocidin, and SCCmec and agr typing. RESULTS: One hundred thirty-one residents (62%) were colonized with S. aureus (MRSA colonization in 86). S. aureus colonization occurred in 80 (76%) of 105 residents with indwelling devices and in 51 (47%) of 108 residents without indwelling devices (P<.001). Of the 86 residents who were colonized with MRSA, nares culture results were positive for only 56 (65%). Residents with devices in place were more likely to be colonized at multiple sites. Eleven different strains of MRSA were identified by pulsed field gel electrophoresis. Seventy-three residents (85%) were colonized with hospital-associated SCCmec II strains, and 8 (9%) were colonized with community associated SCCmec IV strains, 2 of which carried Panton-Valentine leukocidin. CONCLUSIONS: Extranasal colonization with MRSA is common among nursing home residents-particularly among residents with an indwelling device. We documented the emergence of community-associated SCCmec IV MRSA strains in the community nursing home setting in southeastern Michigan. PMID- 18419440 TI - Plasmodium vivax infection during pregnancy: an important problem in need of new solutions. PMID- 18419439 TI - Adverse pregnancy outcomes in an area where multidrug-resistant plasmodium vivax and Plasmodium falciparum infections are endemic. AB - BACKGROUND: Plasmodium falciparum infection exerts a considerable burden on pregnant women, but less is known about the adverse consequences of Plasmodium vivax infection. METHODS: In Papua, Indonesia, where multiple drug resistance to both species has emerged, we conducted a cross-sectional hospital-based study to quantify the risks and consequences of maternal malaria. RESULTS: From April 2004 through December 2006, 3046 pregnant women were enrolled in the study. The prevalence of parasitemia at delivery was 16.8% (432 of 2570 women had infections), with 152 (35.2%) of these 432 infections being associated with fever. The majority of infections were attributable to P. falciparum (250 [57.9%]); 146 (33.8%) of the infections were attributable to P. vivax, and 36 (8.3%) were coinfections with both species. At delivery, P. falciparum infection was associated with severe anemia (hemoglobin concentration, <7 g/dL; odds ratio [OR], 2.8; 95% confidence interval [95% CI], 2.0-4.0) and a 192 g (95% CI, 119 265) reduction in mean birth weight (P<.001). P. vivax infection was associated with an increased risk of moderate anemia (hemoglobin concentration, 7-11 g/dL; OR, 1.8; 95% CI, 1.2-2.9; P=.01) and a 108 g (95% CI, 17.5-199) reduction in mean birth weight (P<.019). Parasitemia was associated with preterm delivery (OR, 1.5; 95% CI, 1.1-2.0; P=.02) and stillbirth (OR, 2.3; 95% CI, 1.3-4.1; P=.007) but was not associated with these outcomes after controlling for the presence of fever and severe anemia, suggesting that malaria increases the risk of preterm delivery and stillbirth through fever and contribution to severe anemia rather than through parasitemia per se. CONCLUSIONS: These observations highlight the need for novel, safe, and effective treatment and prevention strategies against both multidrug-resistant P. falciparum and multidrug-resistant P. vivax infections in pregnant women in areas of mixed endemicity. PMID- 18419441 TI - Doxycycline treatment of Brugia malayi-infected persons reduces microfilaremia and adverse reactions after diethylcarbamazine and albendazole treatment. AB - BACKGROUND: The efficacy of doxycycline for treating the causal agent of human lymphatic filariasis, Brugia malayi, is unknown. Standard treatment with diethylcarbamazine-albendazole is associated with adverse reactions. We assessed whether doxycycline alone or in combination with diethylcarbamazine-albendazole would lead to sustained amicrofilaremia and reduced incidence of adverse reactions. METHODS: A double-blind, randomized, placebo-controlled 6-week field trial of doxycycline treatment (100 mg/day) of 161 persons infected with B. malayi was conducted. Four months after receiving doxycycline (n=119) or placebo (n=42), participants received diethylcarbamazine (6 mg/kg) plus albendazole (400 mg) or a matching placebo. Adverse reactions were assessed 48 and 60 h after administration of diethylcarbamazine-albendazole. Treatment efficacy was evaluated at 2, 4, and 12 months after the initial doxycycline treatment. RESULTS: Four months after beginning doxycycline treatment, Wolbachia loads were reduced by 98%. Doxycycline treatment reduced the prevalence of microfilaremia at 2, 4, and 12 months of follow-up (P<.001 for all time points). At the 1-year follow-up, prevalence was reduced by 77% and 87.5% in patients receiving doxycycline alone or doxycycline plus diethylcarbamazine-albendazole, respectively. In contrast, the reduction of microfilaremia in the group receiving placebo doxycycline plus diethylcarbamazine-albendazole was merely 26.7%. Adverse reactions were lowest in the group receiving doxycycline plus placebo diethylcarbamazine-albendazole and highest in the group receiving placebo doxycycline plus diethylcarbamazine-albendazole. The proportion of persons with high fever and severe adverse reactions was significantly reduced in the group treated with doxycycline plus diethylcarbamazine-albendazole. CONCLUSIONS: A 6 week course of doxycycline, either alone or in combination with diethylcarbamazine-albendazole, leads to a decrease in microfilaremia and reduces adverse reactions to antifilarial treatment in B. malayi-infected persons. PMID- 18419442 TI - A nationally coordinated laboratory system for human avian influenza A (H5N1) in Thailand: program design, analysis, and evaluation. AB - BACKGROUND: The first phase of national surveillance for avian influenza (H5N1) human disease in Thailand occurred over a 4-month period that began on 1 December 2003. Subsequently, a nationally coordinated laboratory system (NCLS) for avian influenza (H5N1) was created to assess population-based surveillance, specimen procurement, case detection, and reporting at the national level. METHODS: We conducted a pre- and postintervention study to evaluate the NCLS designed during the 6-week interval from 1 April through 15 May 2004. During the pre-NCLS period (1 December 2003 through 31 March 2004), 12 cases of human avian influenza (H5N1) were confirmed. During the post-NCLS period (16 May 2004 through 31 December 2006), interventions were implemented for human avian influenza (H5N1) surveillance, case detection, and expedited, computer-based reporting. RESULTS: During the pre- and post-NCLS periods, 777 (85%) of 915 and 10,434 (95%) of 11,042 clinical respiratory specimens, respectively, were adequate for confirmatory testing (P<.001), the median time from procurement to results decreased from 17 days (range, 14-24 days) to 1.8 days (range, 0.25-4 days; P<.001), and the duration of specimen shipment decreased from 46.5 h to 21.1 h (P<.001). Thirteen cases of avian influenza (H5N1) were detected during the 31 month postintervention period. H5N1 reverse-transcriptase polymerase chain reaction and real-time reverse-transcriptase polymerase chain reaction sensitivity was 100% and specificity was 99.8%. CONCLUSIONS: The NCLS exemplifies a systematic approach to national surveillance for avian influenza A (H5N1). This NCLS program in Thailand serves as a model for human avian influenza (H5N1) preparedness that can be adopted or modified for use in other countries. PMID- 18419443 TI - Aerosolized liposomal amphotericin B for the prevention of invasive pulmonary aspergillosis during prolonged neutropenia: a randomized, placebo-controlled trial. AB - BACKGROUND: Invasive pulmonary aspergillosis (IPA) is a significant problem in patients with chemotherapy-induced prolonged neutropenia. Because pulmonary deposition of conidia is the first step in developing IPA, we hypothesized that inhalation of liposomal amphotericin B would prevent IPA. METHODS: We performed a randomized, placebo-controlled trial of patients with hematologic disease with expected neutropenia for >or=10 days. Patients were randomized to receive liposomal amphotericin B or placebo inhalation twice a week, using an adaptive aerosol delivery system, until neutrophil counts increased to >300 cells/mm3. In subsequent neutropenic episodes, the assigned treatment was restarted. The primary end point was the occurrence of IPA according to European Organization for Research and the Treatment of Cancer-Mycoses Study Group definitions. Kaplan Meier curves were compared with log-rank tests for intent-to-treat and on treatment populations. RESULTS: A total of 271 patients were studied during 407 neutropenic episodes. According to the intent-to-treat analysis, 18 of 132 patients in the placebo group developed IPA versus 6 of 139 patients in the liposomal amphotericin B group (odds ratio, 0.26; 95% confidence interval, 0.09 0.72; P=.005). According to the on-treatment analysis, 13 of 97 patients receiving placebo versus 2 of 91 receiving liposomal amphotericin B developed IPA (odds ratio, 0.14; 95% confidence interval, 0.02-0.66; P=.007). Some adverse effects, but none serious, in the liposomal amphotericin B group were reported, most frequently coughing (16 patients vs. 1 patient; P=.002). CONCLUSION: In high risk patients, prophylactic inhalation of liposomal amphotericin B significantly reduced the incidence of IPA. PMID- 18419444 TI - Aerosolized antifungal prophylaxis: the winds of change? PMID- 18419445 TI - Strong correlation between serum aspergillus galactomannan index and outcome of aspergillosis in patients with hematological cancer: clinical and research implications. AB - BACKGROUND: Galactomannan is an Aspergillus-specific polysaccharide released during aspergillosis and is detected by the quantitative serum galactomannan index (GMI) test. Preclinical and preliminary clinical reports have suggested a good correlation between GMI and aspergillosis outcome. METHODS: We reviewed the literature to assess the strength of correlation between GMI and aspergillosis outcome using the kappa correlation coefficient. We included 27 studies that enrolled patients with hematological cancer and proven or probable aspergillosis and that used sequential GMI testing. We examined the 3 following outcomes: survival (survival vs. death), global outcome (survival vs. death [including autopsy findings]), and autopsy outcome (autopsy findings only). RESULTS: Overall, 257 patients fulfilled criteria for proven or probable aspergillosis and were eligible for outcome evaluation. Correlation between GMI (within 40 years of age who have normal calcium levels. The incidence of shingles increased in a linear fashion, from an annual rate of 1.5% among patients with serum calcium levels <10.5 mg/dL to 11% among patients whose calcium levels reached 13 mg/dL (P<.05), a rate that is 6 times greater than that among age-matched historical control individuals (P<.05). PMID- 18419454 TI - Artesunate as a potent antiviral agent in a patient with late drug-resistant cytomegalovirus infection after hematopoietic stem cell transplantation. AB - This is the first report of treatment of cytomegalovirus infection with artesunate, for a stem cell transplant recipient with a newly identified foscarnet-resistant and ganciclovir-resistant DNA polymerase L776M mutation. Artesunate treatment resulted in a 1.7-2.1-log reduction in viral load by treatment day 7, with a viral half-life of 0.9-1.9 days, indicating a highly effective block in viral replication. PMID- 18419455 TI - Abdominal pain and bacterial meningitis in a previously healthy young adult. PMID- 18419456 TI - Vaccination in patients with chronic rheumatic or autoimmune diseases. AB - Patients who have chronic rheumatic or autoimmune diseases, such as rheumatoid arthritis, systemic lupus erythematosus, or vasculitides, show a risk of infection that is at least 2-fold greater than that for healthy individuals. This increased risk is not only a result of the aberrant immunologic reaction itself but also can be attributed to the immunosuppressive therapy required to control the activity of the underlying disease and the associated organ complications. Vaccination is an option for a substantial number of these infections. In this context, pneumococcal and influenza vaccines are the best evaluated and are recommended by standard vaccination guidelines. Some studies have found mildly impaired immune responses to vaccines among patients receiving long-term immunosuppressive therapy, but postvaccination antibody titers are usually sufficient to provide protection for the majority of immunized individuals. The accumulated data on the safety and effectiveness of vaccines warrant immunization with the majority of vaccines for patients with chronic autoimmune or rheumatic diseases, especially vaccination against influenza and pneumococci. Vaccination protocols for this population should be better implemented in daily clinical practice. PMID- 18419458 TI - Do Vgamma2Vdelta2 T cells influence HIV disease progression? PMID- 18419457 TI - Association between Vgamma2Vdelta2 T cells and disease progression after infection with closely related strains of HIV in China. AB - BACKGROUND: Human immunodeficiency virus (HIV) infection and disease are accompanied by decreases in the absolute number and function of Vgamma2Vdelta2 T cells, suggesting that this subset of cells may play an important role in controlling disease. We performed a cross-sectional study involving HIV-infected former blood donors and assessed the association between Vgamma2Vdelta2 T cells and markers of disease progression. METHODS: Changes in Vgamma2Vdelta2 T cell count and function were compared between HIV-infected individuals and healthy blood donors using the Mann-Whitney U test. The relationships between Vgamma2Vdelta2 T cell count, plasma viral load, and CD4 T cell count were analyzed using the Spearman correlation. RESULTS: We found significant positive correlations between CD4 T cell count and both total Vgamma2Vdelta2 T cell count (P<.001) and functional (isopentenyl pyrophosphate-responsive) Vgamma2Vdelta2 T cell count (P<.001). We found significant reverse correlations between viral load and both total Vgamma2Vdelta2 T cell count (P<.05) and functional Vgamma2Vdelta2 T cell count (P<.05). CONCLUSIONS: The association of Vgamma2Vdelta2 T cells with disease progression in 146 HIV-infected participants supports the view that intact Vgamma2Vdelta2 T cell populations are important for controlling HIV disease. PMID- 18419459 TI - Epstein-Barr virus-associated central nervous system lymphoproliferative disease in a patient with acquired immunodeficiency syndrome responsive to highly active antiretroviral therapy. AB - A 20-year-old man with acquired immunodeficiency syndrome (AIDS) and central nervous system (CNS) lymphoproliferative disease experienced improvement with highly active antiretroviral therapy (HAART) without radiation therapy. Our experience highlights the importance of biopsy in evaluating multifocal radiographic CNS lesions and the central role of HAART in treating AIDS-related CNS disease. PMID- 18419461 TI - Aspirin use in infective endocarditis. PMID- 18419462 TI - Determination of the incidence of tuberculosis in low-income countries. PMID- 18419464 TI - Vancomycin minimum inhibitory concentration as a predictor of mortality in methicillin-resistant Staphylococcus aureus bacteremia: a second look. PMID- 18419465 TI - Shock as a covariate in a study of treatment of methicillin-resistant Staphylococcus aureus bacteremia. PMID- 18419467 TI - Hospital tap water as a source of stenotrophomonas maltophilia infection. PMID- 18419469 TI - Vaccines to prevent systemic mycoses: holy grails meet translational realities. PMID- 18419470 TI - Persistence of measles, mumps, and rubella antibodies in an MMR-vaccinated cohort: a 20-year follow-up. AB - BACKGROUND: The persistence of antibodies against measles, mumps, and rubella induced by the measles-mumps-rubella (MMR) vaccine and the kinetics of antibody decline after the second MMR vaccine dose were studied in the same cohort for 20 years. METHODS: Measles, mumps, and rubella antibodies were measured by enzyme immunoassay in 20-year follow-up serum samples (n= 183) of twice-vaccinated individuals, and measles antibodies were also measured in oral fluids (n = 177). Antibody decay was determined in a group (n = 58) with subsequent samples collected 1, 8, and 15 years after the second MMR dose. RESULTS: In total, 95%, 74%, and 100% of 183 vaccinees were still seropositive for measles, mumps, and rubella, respectively, and 85% of 177 vaccinees had measurable measles antibodies in their oral fluids. The antibody levels declined significantly after the second dose, but subsequently the rate of decline was slower. CONCLUSIONS: A high rate of seropositivity was found 20 years after the first MMR dose, particularly for rubella and measles. Our results show that MMR vaccine-induced antibodies wane significantly after the second dose. According to epidemiological data, the protection induced by MMR vaccination in Finland seems to persist at least until early adulthood. However, the situation requires constant vigilance. PMID- 18419472 TI - Catecholamine inotrope resuscitation of antibiotic-damaged staphylococci and its blockade by specific receptor antagonists. AB - The increasing use of antibiotic-coated catheters, such as those containing rifampin or minocycline, has led to a decrease in catheter colonization by staphylococci but not to a decrease in the incidence of catheter-related bloodstream infection (BSI). Because catheters are used for the administration of catecholamine inotropes to maintain cardiac function, we examined whether 2 commonly employed inotropes, dopamine and norepinephrine, could affect bacterial viability after exposure to rifampin and minocycline. Rifampin inhibition and minocycline inhibition of staphylococcal growth could be reversed by exposure to dopamine or norepinephrine as a result, in part, of catecholamine-mediated increased provision of host-sequestered iron. The simultaneous addition of inotropes with an antibiotic did not affect antibiotic susceptibility. Inotrope induced growth in bacteria previously exposed to antibiotics was blocked by the inclusion in culture media of specific catecholamine-receptor antagonists. Considered collectively, these results provide a mechanistic basis for understanding how host-related factors, such as inotrope-based therapeutics, may influence the recovery of antibiotic-stressed bacteria in clinical settings. PMID- 18419471 TI - Antibody titer threshold predicts anti-candidal vaccine efficacy even though the mechanism of protection is induction of cell-mediated immunity. AB - We previously reported that vaccination with Freund's adjuvant plus the recombinant N-terminus of the candidal adhesin, Als3p (rAls3p-N), protects mice from disseminated candidiasis. Here we report that the rAls3p-N vaccine is effective when combined with aluminum hydroxide adjuvant. Antibody titers of > or =1:6400 accurately predicted protection from infection. Nevertheless, neither B lymphocytes nor serum from immunized animals transferred protection to vaccine naive animals. In contrast, CD3(+), CD4(+), or CD8(+) T lymphocytes from immunized animals transferred protection, and the vaccine was efficacious in IL-4 deficient mice but not in IFN-gamma-deficient mice. These data have significant implications for the development and interpretation of vaccine surrogate markers. PMID- 18419473 TI - Rho kinase inhibition in severe malaria: thwarting parasite-induced collateral damage to endothelia. AB - Acute clinical manifestations of falciparum malaria, such as multiorgan failure and cerebral malaria, occur unpredictably and lead to coma and death within hours if left untreated. Despite the emergency administration of effective antimalarial drugs, 15%-20% of patients die. Other therapeutic approaches are therefore urgently needed. There is increasing evidence that endothelial changes play a key role in the pathogenesis of severe malaria. We therefore used coculture models to study interactions between infected erythrocytes and endothelium. We found that adhesion of Plasmodium falciparum to endothelial cells in vitro activated the Rho kinase signaling pathway, which is strongly involved in various vascular diseases. When added concomitantly with parasites, the Rho kinase inhibitor fasudil (HA-1077), a drug already in clinical use, decreased both NF-kappaB activation and endothelial cell apoptosis. Fasudil also helped to restore endothelial barrier integrity after P. falciparum adhesion. Rho kinase inhibition thus appears to be a promising adjunctive therapeutic approach to the management of severe human malaria. PMID- 18419475 TI - Candida arteritis in patients who have not received organ transplants: case report and review of the literature. AB - Fungal arteritis is a rare entity, associated with significant morbidity and mortality, that typically involves graft arteries in solid organ transplant recipients. Here, we report the first case of Candida carotid arteritis and review 22 other cases of Candida arteritis reported since 1966 in patients who have not received transplants. Most patients had serious underlying conditions. All cases were anatomically characterized by pseudoaneurysm formation. Patients presented with fever (n=7), local pain (n=13), and an expanding pulsatile mass (n=4), with subsequent rupture and hemorrhage (n=7). Evidence of Candida colonization or infection was present in 15 patients before arteritis occurred. Treatment typically included a combined surgical and antifungal approach. The outcome was favorable in 11 patients, but follow-up was limited. A high index of suspicion, early diagnosis, and prompt antifungal and surgical treatment seem crucial to efforts to avoid life-threatening arterial rupture and hemorrhage. PMID- 18419476 TI - Lactational amenorrhea as a risk factor for group a streptococcal vaginitis. AB - We report a case of Streptococcus pyogenes, beta-hemolytic Streptococcus, Lancefield group A vulvovaginitis in an otherwise healthy adult female patient experiencing lactational amenorrhea. Group A streptococcal infection is the infective cause of vulvovaginitis in 21% of prepubescent girls, but it is an uncommon cause of vulvovaginitis in adults. Group A streptococcal vulvovaginitis is frequently associated with households that have had a recent outbreak of respiratory or dermal infection. The case described here appears to be unusual in that it was sexually transmitted, and the lack of estrogen associated with anovualtion may have been a predisposing factor for this unusual sexually transmitted disease. PMID- 18419477 TI - Invasive pulmonary infection due to Trichoderma longibrachiatum mimicking invasive Aspergillosis in a neutropenic patient successfully treated with voriconazole combined with caspofungin. AB - Trichoderma longibrachiatum, a filamentous fungus, was recently described as an emerging pathogen in immunocompromised patients. Here, we report the first case, to our knowledge, of isolated invasive lung infection by T. longibrachiatum in a patient with hematologic malignancy. The infection mimicked invasive pulmonary aspergillosis and was successfully treated with a combination of voriconazole and caspofungin. PMID- 18419478 TI - Multimer staining of cytomegalovirus phosphoprotein 65-specific T cells for diagnosis and therapeutic purposes: a comparative study. AB - BACKGROUND: Cytomegalovirus (CMV) disease represents a serious complication after allogeneic peripheral blood stem cell (PBSC) transplantation. If possible, stem cell donors for transplantation are selected on the basis of their CMV serostatus. However, the cytomegalovirus-specific immune status can be further characterized by measuring CMV phosphoprotein 65-specific CD8(+) T cell frequencies using tetramers, pentamers, and streptamers. We therefore investigated the specificity and sensitivity of all 3 methods and compared the results to patient serostatus. METHODS: Twenty-three samples from CMV seropositive healthy volunteers and 15 samples from CMV-seropositive patients before and after allogeneic PBSC transplantation were stained with tetramers, pentamers, or streptamers and analyzed by flow cytometry. RESULTS: Similar frequencies of CD8(+) and multimer(+) T cells could be measured by all 3 multimer technologies. The lowest background signals (< or =0.02%) were obtained using tetramer technology. Frequencies of 0.19%-2.48% of CMV phosphoprotein 65 495-503 specific CD8(+) T cells were detected in healthy volunteers. Antigen-specific T cells were detected in only 11 (48%) of 23 seropositive healthy volunteers. CMV antigenemia before day 100 after allogeneic PBSC transplantation occurred in 2 of 3 patients without any specific T cells. CONCLUSION: These findings demonstrate the power of multimer staining and a certain limitation of serologic testing to define appropriate donors for transplantation. Therefore, whenever possible, CMV seropositive donors of transplants to seropositive recipients should be screened for their CD8(+) T cell frequency. All 3 multimer technologies can be used, yielding similar results. The streptamer technology additionally offers the advantage of selecting CMV phosphoprotein 65-specific CD8(+) T cells at the good manufacturing practice level for adoptive T cell transfer. PMID- 18419480 TI - The case for vancomycin as the preferred drug for treatment of Clostridium difficile infection. PMID- 18419481 TI - Vancomycin for the treatment of Clostridium difficile Infection: for whom is this expensive bullet really magic? AB - The epidemiology, clinical severity, and case-fatality ratio of Clostridium difficile infection (CDI) changed dramatically with the emergence of a toxin hyperproducing strain (BI/NAP1/027) in North America and Europe in 2000. For the treatment of CDI, metronidazole and vancomycin remain the 2 most commonly used drugs. The 3 randomized controlled trials published thus far, as well as the upcoming tolevamer trial, use intermediate outcomes, rather than the outcomes that now preoccupy clinicians: the frequency of complications or recurrence. The major advantage of metronidazole is its low price. The major advantage of orally administered vancomycin is its more favorable pharmacokinetics. Facilitating vancomycin-resistant enterococci colonization and/or infection is a potential drawback of both drugs. Pending the development of a prospectively validated scoring system, members of the Infectious Diseases Society of America/Society for Healthcare Epidemiology of America expert committee will define severe CDI as present in any patient with a leukocyte count > or =15,000 cells/mm(3) or a creatinine level increased by > or =50% from baseline. For patients with mild-to moderate CDI (defined by a leukocyte count <15000 cells/mm(3) and a creatinine level <1.5 times the baseline value), there is no evidence that treatment with vancomycin is superior to treatment with metronidazole (even for intermediate outcomes), and metronidazole therapy should be preferred. For patients with severe CDI who are not infected with BI/NAP1/027, there is reasonable evidence that the better pharmacokinetics of vancomycin translate into a lower probability of complications. For those patients who are infected with BI/NAP1/027, the superiority of vancomycin therapy remains to be proven. In practice, because it is not yet possible to rapidly type the strains, all patients with severe CDI should be treated with vancomycin. Future trials should use complicated CDI and recurrences as their primary outcomes. PMID- 18419483 TI - Monotherapy or combination therapy for hospitalized patients with community acquired pneumonia: not yet the end of the story? PMID- 18419482 TI - Moxifloxacin monotherapy is effective in hospitalized patients with community acquired pneumonia: the MOTIV study--a randomized clinical trial. AB - BACKGROUND: The aim of this study was to show that sequential intravenous and oral moxifloxacin monotherapy (400 mg once per day) is as efficacious and safe as a combination regimen (intravenous ceftriaxone, 2 g once per day, plus sequential intravenous and oral levofloxacin, 500 mg twice per day) in patients hospitalized with community-acquired pneumonia. METHODS: We conducted a prospective, multicenter, randomized, double-blind noninferiority trial. Patients with a Pneumonia Severity Index (PSI) of III-V were stratified on the basis of PSI risk class before randomization. The primary efficacy end point was clinical response at test of cure (4-14 days after the completion of treatment). Secondary efficacy end points were clinical and bacteriological response at end of treatment (days 7 14) and at follow-up assessment (21-28 days after the end of treatment), overall mortality, and mortality attributable to pneumonia. RESULTS: Seven hundred thirty three patients were enrolled in the study (368 in the moxifloxacin arm and 365 in the comparator arm); 49% had a PSI of IV, and 10% had a PSI of V. Of 569 patients (291 in the moxifloxacin arm and 278 in the comparator arm) valid for per protocol analysis, the overall clinical cure rates at test of cure were 86.9% for moxifloxacin and 89.9% for the comparator regimen (95% confidence interval, -8.1% to 2.2%). Bacteriological success at test of cure was 83.3% for moxifloxacin and 85.1% for the comparator regimen (95% confidence interval, -15.4% to 11.8%). There were no significant differences between moxifloxacin and comparator treatments in the incidence of treatment-emergent adverse events or in mortality. CONCLUSIONS: Monotherapy with sequential intravenous/oral moxifloxacin was noninferior to treatment with ceftriaxone plus levofloxacin combination therapy in patients with community-acquired pneumonia who required hospitalization. PMID- 18419484 TI - The etiology of community-acquired pneumonia in Australia: why penicillin plus doxycycline or a macrolide is the most appropriate therapy. AB - BACKGROUND: Available data on the etiology of community-acquired pneumonia (CAP) in Australia are very limited. Local treatment guidelines promote the use of combination therapy with agents such as penicillin or amoxycillin combined with either doxycycline or a macrolide. METHODS: The Australian CAP Study (ACAPS) was a prospective, multicenter study of 885 episodes of CAP in which all patients underwent detailed assessment for bacterial and viral pathogens (cultures, urinary antigen testing, serological methods, and polymerase chain reaction). Antibiotic agents and relevant clinical outcomes were recorded. RESULTS: The etiology was identified in 404 (45.6%) of 885 episodes, with the most frequent causes being Streptococcus pneumoniae (14%), Mycoplasma pneumoniae (9%), and respiratory viruses (15%; influenza, picornavirus, respiratory syncytial virus, parainfluenza virus, and adenovirus). Antibiotic-resistant pathogens were rare: only 5.4% of patients had an infection for which therapy with penicillin plus doxycycline would potentially fail. Concordance with local antibiotic recommendations was high (82.4%), with the most commonly prescribed regimens being a penicillin plus either doxycycline or a macrolide (55.8%) or ceftriaxone plus either doxycycline or a macrolide (36.8%). The 30-day mortality rate was 5.6% (50 of 885 episodes), and mechanical ventilation or vasopressor support were required in 94 episodes (10.6%). Outcomes were not compromised by receipt of narrower-spectrum beta-lactams, and they did not differ on the basis of whether a pathogen was identified. CONCLUSIONS: The vast majority of patients with CAP can be treated successfully with narrow-spectrum beta-lactam treatment, such as penicillin combined with doxycycline or a macrolide. Greater use of such therapy could potentially reduce the emergence of antibiotic resistance among common bacterial pathogens. PMID- 18419485 TI - Treatment of community-acquired pneumonia Down Under versus the United States: is it really that different? PMID- 18419486 TI - Clinical and virological characteristics of 15 patients with chronic active Epstein-Barr virus infection treated with hematopoietic stem cell transplantation. AB - BACKGROUND: Chronic active Epstein-Barr virus (EBV) infection is characterized by recurrent infectious mononucleosis-like symptoms, and infected patients have high viral loads in their peripheral blood. Standard therapy for the disease has not yet been established. Recently, hematopoietic stem cell transplantation (HSCT) has been introduced and has the potential to become a standard treatment, although guidelines for HSCT to treat chronic active EBV infection have not yet been proposed. METHODS: Fifteen patients were retrospectively analyzed, both clinically and virologically, to investigate the factors associated with prognosis of chronic active EBV infection treated with HSCT. RESULTS: After HSCT, 7 patients died after survival periods that ranged from 1 to 16 months (mean duration of survival after HSCT, 5 months). Three patients were considered to have died of transplantation-related complications. The duration between infection onset and diagnosis was significantly longer in patients who died than in those who survived. Five of the 7 patients who died experienced > or =3 life threatening complications. The plasma concentrations of interferon-gamma, interleukin-10, thrombomodulin, and soluble E-selectin did not differ significantly between the groups of patients. With regard to sequence variations in the EBV latent membrane protein 1 gene, no specific patterns were found in the patients who died. Importantly, the plasma EBV load at diagnosis was significantly higher in patients who died than in living patients. Moreover, plasma viral load was shown to be an important factor to monitor during follow-up for patients after HSCT. CONCLUSIONS: The number of life-threatening complications and plasma viral load are indicative of the stage of disease progression and may be useful factors for predicting the outcome of HSCT. PMID- 18419487 TI - Clinical and virological characteristics of patients with chronic active Epstein Barr virus infection treated with hematopoietic stem cell transplantation: insights and questions. PMID- 18419488 TI - Mortality associated with bloodstream infection after coronary artery bypass surgery. AB - BACKGROUND: Mortality attributable to bloodstream infection (BSI) is still controversial. We studied the impact of BSI on mortality after coronary artery bypass surgery, including the specific impact of different etiologic organisms. METHODS: Our cohort consisted of 4515 patients who underwent coronary artery bypass procedures at a university hospital from 1996 through 2004. We used Society of Thoracic Surgery data supplemented with laboratory and infection control data. Mortality dates were identified using Society of Thoracic Surgery data and the Social Security Death Index. BSI within 90 days after surgery was defined by a positive blood culture result. Cox proportional hazards and propensity score models were used to analyze the association between BSI and mortality. RESULTS: Patients with BSI had a 4.2-fold increased risk of death (95% confidence interval [CI], 3.0-5.9) 2-90 days after coronary artery bypass surgery, compared with uninfected patients. The risk of death was higher among patients with BSI due to gram-negative bacteria (hazard ratio [HR], 6.8; 95% CI, 3.9-12.0) and BSI due to Staphylococcus aureus (HR, 7.2; 95% CI, 3.3-15.7) and lowest among patients with BSI caused by gram-positive bacteria other than S. aureus (HR, 2.2; 95% CI, 1.1-4.6). The risk of death was highest among patients who developed BSI but had the lowest likelihood of infection (HR, 10.0; 95% CI, 3.5-28.8) and was lowest among patients who developed BSI but had the highest likelihood of infection (HR, 2.3; 95% CI, 1.2-4.6). CONCLUSIONS: BSIs due to gram negative bacteria and BSIs due to S. aureus contributed significantly to mortality. Mortality attributable to BSI was highest among patients predicted to be least likely to develop infection and was lowest among severely ill patients who were most likely to develop infection. BSI appears to be an important contributor to death after coronary artery bypass surgery, particularly among the healthiest patients. PMID- 18419491 TI - The deadly toll of invasive methicillin-resistant Staphylococcus aureus infection in community hospitals. AB - INTRODUCTION: Data regarding the epidemiology, treatment, and outcomes of methicillin-resistant Staphylococcus aureus (MRSA) infections in rural and community hospitals are limited. METHODS: This cohort study was conducted at 1 tertiary care hospital and 8 community hospitals in the southeastern United States. Patients with a surgical site infection (SSI) and/or bacteremia due to MRSA were prospectively identified at each study hospital during the period 1994 2003. RESULTS: A total of 129 patients with SSI and 564 patients with bacteremia due to MRSA were identified. Only 57 patients with SSI (44.2%) received antibiotics active against MRSA on the initial date of diagnosis; only 95 (73.6%) received an agent active against MRSA by day 7 after diagnosis of SSI due to MRSA. Ninety-five patients with SSI due to MRSA (73.6%) were readmitted to the hospital within 90 days after their original surgery. The 1-year mortality rate among patients with SSI due to MRSA was 22%. Inadequate therapy was also commonly given to patients with bacteremia: only 216 (38.3%) received antibiotics active against MRSA on the initial day of infection, and only 383 (67.9%) received an agent active against MRSA by day 7 after diagnosis. Approximately one-third of patients with bloodstream infection died during their initial hospitalization. Patients hospitalized in community hospitals were less likely to receive effective antimicrobial therapy on both the day of infection and within 7 days after infection, compared with patients in the tertiary care hospital. CONCLUSION: Inadequate therapy is commonly administered after diagnosis of SSI and bacteremia due to MRSA in patients in community hospitals. PMID- 18419489 TI - Infections in 252 patients with common variable immunodeficiency. AB - BACKGROUND: Common variable immunodeficiency is characterized by recurrent infections and defective immunoglobulin production. METHODS: The DEFI French national study prospectively enrolled adult patients with primary hypogammaglobulinemia. Clinical events before inclusion were retrospectively analyzed at that time. RESULTS: From April 2004 through April 2007, 341 patients were enrolled, 252 of whom had received a diagnosis of common variable immunodeficiency; of those, 110 were male, 142 were female, and 228 were white. The median age at first symptoms was 19 years. The median age at common variable immunodeficiency diagnosis was 33.9 years. The median delay for diagnosis was 15.6 years for the 138 patients with initial symptoms before 1990 and 2.9 years for the 114 patients with initial symptoms from 1990 to the time of the study. The most frequent initial symptoms were upper respiratory tract infections: bronchitis (in 38% of patients), sinusitis (36%), pneumonia (31%), and/or bronchiectasis (14%). Overall, 240 patients had respiratory symptoms. Pneumonia was reported in 147 patients; Streptococcus pneumoniae and Haemophilus influenzae were documented in 46 and 17 cases, respectively. Recurrent or chronic diarrhea was reported in 118 patients. Giardia (35 cases), Salmonella (19), and Campylobacter (19) infections were more frequent in patients with undetectable serum immunoglobulin A (P<.001). Sixteen patients developed opportunistic infections. Persistent infections and requirement for antibiotics despite immunoglobulin substitution correlated with severe defect of memory switched B cells (P=.003) but not with immunoglobulin G trough levels (P=.55). CONCLUSION: Although reduced within the past decade, the delay of diagnosis of common variable immunodeficiency remains unacceptable. Recurrence of upper respiratory tract infection or pneumonia should lead to systematic evaluation of serum immunoglobulin. PMID- 18419490 TI - Severe eczema vaccinatum in a household contact of a smallpox vaccinee. AB - BACKGROUND: We report the first confirmed case of eczema vaccinatum in the United States related to smallpox vaccination since routine vaccination was discontinued in 1972. A 28-month-old child with refractory atopic dermatitis developed eczema vaccinatum after exposure to his father, a member of the US military who had recently received smallpox vaccine. The father had a history of inactive eczema but reportedly reacted normally to the vaccine. The child's mother also developed contact vaccinia infection. METHODS: Treatment of the child included vaccinia immune globulin administered intravenously, used for the first time in a pediatric patient; cidofovir, never previously used for human vaccinia infection; and ST-246, an investigational agent being studied for the treatment of orthopoxvirus infection. Serological response to vaccinia virus and viral DNA levels, correlated with clinical events, were utilized to monitor the course of disease and to guide therapy. Burn patient-type management was required, including skin grafts. RESULTS: The child was discharged from the hospital after 48 days and has recovered with no apparent systemic sequelae or significant scarring. CONCLUSION: This case illustrates the need for careful screening prior to administration of smallpox vaccine and awareness by clinicians of the ongoing vaccination program and the potential risk for severe adverse events related to vaccinia virus. PMID- 18419492 TI - Culture-Negative intracerebral abscesses in children and adolescents from Streptococcus anginosus group infection: a case series. AB - We report the use of 16S ribosomal RNA gene amplification and sequencing to diagnose culture-negative intracerebral abscesses in younger patients. These 3 cases demonstrate the optimal application of gene sequencing from direct specimens for patients with negative culture results compromised by antibacterial therapy but histories highly suggestive of acute bacterial infection. PMID- 18419493 TI - An elderly woman with a diffuse annular eruption. PMID- 18419494 TI - The interaction between nutrition and infection. AB - Infection and malnutrition have always been intricately linked. Malnutrition is the primary cause of immunodeficiency worldwide, and we are learning more and more about the pathogenesis of this interaction. Five infectious diseases account for more than one-half of all deaths in children aged <5 years, most of whom are undernourished. Micronutrient deficiencies have effects such as poor growth, impaired intellect, and increased mortality and susceptibility to infection. The worldwide magnitude of parasite infection is enormous. It is understood that parasites may lead to malnutrition, but the extent to which malnutrition causes increased parasite infestation is not known; thus, the conditions need to be addressed together. Nutritional deficiencies associated with pregnancy are associated with poor immune response to infection. Because this immune deficiency is partially compensated by breast-feeding, this is the single best way to protect infants from infection. Malnutrition and nutritional alterations, common complications of human immunodeficiency virus infection, include disorders of food intake, nutrient absorption, and intermediary metabolism and play a significant and independent role in morbidity and mortality. The 21st century provides new information and new challenges. With new technologies and political changes, it is hoped that a healthier, more disease-free, and better-nourished population will emerge. PMID- 18419495 TI - Prevalence of HIV-1 drug resistance after failure of a first highly active antiretroviral therapy regimen in KwaZulu Natal, South Africa. AB - BACKGROUND: Emergence of human immunodeficiency virus type 1 (HIV-1) drug resistance may limit the benefits of antiretroviral therapy in resource-limited settings. The prevalence of resistance was assessed among patients from KwaZulu Natal, South Africa, following failure of their first highly active antiretroviral therapy (HAART) regimen. METHODS: Genotypic resistance testing was performed on plasma virus samples from patients who experienced virologic failure of their first HAART regimen at 2 clinics in KwaZulu Natal. Clinical and demographic data were obtained from medical records. Regression analysis was performed to determine factors associated with > or =1 significant drug resistance mutation. RESULTS: From January 2005 through August 2006, a total of 124 antiretroviral-treated adults who experienced virologic failure were enrolled. The predominant subtype was HIV-1C. Virus samples from 83.5% of participants carried > or =1 significant drug resistance mutation. Dual-class drug-resistant virus was present in 64.3% of participants, and 2.6% had virus with triple-class drug resistance. The most common mutation was M184V/I (64.3% of patients); K103N was present in virus from 51.3%, and V106M was present in virus from 19.1%. Thymidine analog resistance mutations were found in virus from 32.2% of patients, and protease resistance mutations were found in virus from 4.4%. CONCLUSIONS: Antiretroviral drug-resistant virus was detected in >80% of South African patients who experienced failure of a first HAART regimen. Patterns of drug resistance reflected drugs used in first-line regimens and viral subtype. Continued surveillance of resistance patterns is warranted to guide selection of second-line regimens. PMID- 18419496 TI - Running with scissors: using antiretroviral therapy without monitoring viral load. PMID- 18419498 TI - Evaluating adherence to highly active antiretroviral therapy with use of pill counts and viral load measurement in the drug resources enhancement against AIDS and malnutrition program in Mozambique. AB - BACKGROUND: Maintaining treatment adherence among the growing number of patients receiving antiretroviral treatment in Africa is a dramatic challenge. The objective of our study was to explore the results of a computerized pill count method and to test the validity, sensitivity, and specificity of this method with respect to viral load measurement in an African setting. METHODS: We performed a prospective, observational study involving patients who received first-line highly active antiretroviral therapy in Mozambique from 1 April 2005 through 31 March 2006. Enrolled patients had received treatment for at least 3 months before the study. For defining treatment adherence levels, pill counts were used, and the results were analyzed with viral load measurements at the end of the observation period. RESULTS: The study involved 531 participants. During the 12 months of observation, 137 patients left the program or discontinued first-line therapy. Of the remaining 394 patients, 284 (72.1%) had >95% treatment adherence; of those 284 patients, 274 (96.5%) had a final viral load <1000 copies/mL. A Cox proportional hazards analysis revealed that the relationship between >95% treatment adherence and the final viral load was closer than that between >90% treatment adherence and viral load. CONCLUSIONS: Treatment adherence >95% maximizes the results of the nonnucleoside reverse-transcriptase inhibitor-based regimen. The pill count method appears to be a reliable and economic tool for monitoring treatment adherence in resource-limited settings. PMID- 18419499 TI - The impact of HIV tropism on decreases in CD4 cell count, clinical progression, and subsequent response to a first antiretroviral therapy regimen. AB - BACKGROUND: Human immunodeficiency virus (HIV) uses 2 distinct chemokine receptors, CCR5 (R5) or CXCR4 (X4), during entry. Viruses may be R5 tropic, X4 tropic, or dual/mixed (D/M) tropic. R5-tropic virus predominates at high CD4 cell counts, with the number of X4-tropic strains increasing as CD4 cell count decreases. METHODS: We investigated the relationship between tropism and decreases in CD4 cell count before antiretroviral therapy initiation, the frequency of clinical events, and responses to antiretroviral therapy in a cohort of treatment-naive patients. RESULTS: Four hundred two treatment-naive patients underwent tropism determination; 326 harbored R5-tropic virus, and 76 harbored X4 or D/M-tropic virus. After adjustment for baseline characteristics, the rate of decrease in CD4 cell count was significantly greater in patients infected with X4 or D/M-tropic virus at 12 months (P=.026). Two hundred twenty-nine individuals infected with R5-tropic virus and 60 individuals infected with X4- or D/M-tropic virus commenced antiretroviral therapy between tropism testing and the time of data analysis. Time to viral suppression and the proportion of patients achieving viral suppression were similar at 6, 12, and 24 months. CD4 cell count increases were similar. Clinical events were significantly more common in the group infected with X4- or D/M-tropic virus. Multivariate analysis demonstrated a relative risk of experiencing a clinical event of 2.56 (95% confidence interval, 1.37-4.76; P=.003) among patients infected with X4- or D/M-tropic virus. CONCLUSIONS: The presence of D/M- or X4-tropic virus has a deleterious effect on CD4 cell count decrease and risk of clinical disease. Response to standard antiretroviral therapy is not affected by viral tropism. PMID- 18419501 TI - Numbers needed to treat with posaconazole prophylaxis to prevent invasive fungal infection and death. PMID- 18419503 TI - The effect of gastric Acid on the absorption of posaconazole. PMID- 18419504 TI - Antiviral therapy and outcomes in hospitalized patients with influenza. PMID- 18419506 TI - Antiretroviral drug resistance among antiretroviral-naive individuals with HIV infection of unknown duration in Thailand. PMID- 18419497 TI - Plasma drug concentrations and virologic evaluations after stopping treatment with nonnucleoside reverse-transcriptase inhibitors in HIV type 1-infected children. AB - BACKGROUND: The optimum strategy for stopping treatment with drugs that have different half-lives in a combination regimen to minimize the risk of selecting drug-resistant viruses remains unknown. We evaluated drug concentrations in plasma, human immunodeficiency virus (HIV) load, and development of drug resistance after a planned treatment interruption of a nonnucleoside reverse transcriptase inhibitor (NNRTI)-containing regimen in HIV type 1-infected children. METHODS: Children with viral loads <50 copies/mL and CD4 cell percentages > or =30% (for children aged 2-6 years) or CD4 cell percentages > or =25% and CD4 cell counts > or =500 cells/microL (for children aged 7-15 years) were randomized to either a planned treatment interruption or to continuous therapy. In the planned treatment interruption arm, either (1) treatment with nevirapine or efavirenz was stopped, and treatment with the remaining drugs was continued for 7-14 days, or (2) nevirapine or efavirenz were replaced by a protease inhibitor, and all drugs were stopped after 7-14 days. Sampling for determination of plasma drug concentrations, measurement of viral load, and drug resistance testing was scheduled at day 0, day 7 (drug concentrations only), day 14, and day 28 after interruption of treatment with an NNRTI. RESULTS: Treatment with an NNRTI was interrupted for 35 children (20 were receiving nevirapine, and 15 were receiving efavirenz). Median time from NNRTI cessation to stopping all drugs was 9 days (range, 6-15 days) for nevirapine and 14 days (range, 6-18 days) for efavirenz. At 7 days, 1 (5%) of 19 and 4 (50%) of 8 children had detectable nevirapine and efavirenz concentrations, respectively; efavirenz remained detectable in 3 (25%) of 12 children at 14 days. At 14 days, viral load was > or =50 copies/mL in 6 of 16 children interrupting treatment with nevirapine (range, 52-7000 copies/mL) and in 2 of 12 children interrupting treatment with efavirenz (range, 120-1600 copies/mL). No new NNRTI mutations were observed. CONCLUSIONS: In children with virological suppression who experienced interruption of treatment with an NNRTI, staggered or replacement stopping strategies for a median of 9 days for nevirapine and 14 days for efavirenz were not associated with the selection of NNRTI resistance mutations. PMID- 18419507 TI - Use of blood culture vials and nucleic acid amplification for the diagnosis of pediatric septic arthritis. PMID- 18419518 TI - Comparative methods with sampling error and within-species variation: contrasts revisited and revised. AB - Comparative methods analyses have usually assumed that the species phenotypes are the true means for those species. In most analyses, the actual values used are means of samples of modest size. The covariances of contrasts then involve both the covariance of evolutionary changes and a fraction of the within-species phenotypic covariance, the fraction depending on the sample size for that species. Ives et al. have shown how to analyze data in this case when the within species phenotypic covariances are known. The present model allows them to be unknown and to be estimated from the data. A multivariate normal statistical model is used for multiple characters in samples of finite size from species related by a known phylogeny, under the usual Brownian motion model of change and with equal within-species phenotypic covariances. Contrasts in each character can be obtained both between individuals within a species and between species. Each contrast can be taken for all of the characters. These sets of contrasts, each the same contrast taken for different characters, are independent. The within-set covariances are unequal and depend on the unknown true covariance matrices. An expectation-maximization algorithm is derived for making a reduced maximum likelihood estimate of the covariances of evolutionary change and the within species phenotypic covariances. It is available in the Contrast program of the PHYLIP package. Computer simulations show that the covariances are biased when the finiteness of sample size is not taken into account and that using the present model corrects the bias. Sampling variation reduces the power of inference of covariation in evolution of different characters. An extension of this method to incorporate estimates of additive genetic covariances from a simple genetic experiment is also discussed. PMID- 18419519 TI - Individual variation in migration speed of upriver-migrating sockeye salmon in the Fraser River in relation to their physiological and energetic status at marine approach. AB - Little research has examined individual variation in migration speeds of Pacific salmon (Oncorhynchus spp.) in natural river systems or attempted to link migratory behavior with physiological and energetic status on a large spatial scale in the wild. As a model, we used three stocks of summer-run sockeye salmon (Oncorhynchus nerka) from the Fraser River watershed, British Columbia, to test the hypothesis that individual variation in migration speed is determined by a combination of environmental factors (i.e., water temperature), intrinsic biological differences (sex and population), and physiological and energetic condition. Before the freshwater portion of the migration, sockeye salmon (Quesnel, Chilcotin, and Nechako stock complexes) were captured in Johnstone Strait ( approximately 215 km from river entry), gastrically implanted with radio transmitters, and sampled for blood, gill tissue, and energetic status before release. Analyses focused solely on individuals that successfully reached natal subwatersheds. Migration speeds were assessed by an extensive radiotelemetry array. Individuals from the stock complex that migrated the longest distance (Nechako) traveled at speeds slower than those of other stock complexes. Females traveled slower than males. An elevated energetic status of fish in the ocean was negatively correlated with migration speed in most river segments. During the transition from the ocean to the river, migration speed was negatively correlated with mean maximum water temperature; however, for the majority of river segments, it was positively correlated with migration speed. Physiological status measured in the ocean did not explain among-individual variability in river migration speeds. Collectively, these findings suggest that there could be extensive variation in migration behavior among individuals, sexes, and populations and that physiological condition in the ocean explained little of this variation relative to in-river environmental conditions and energetic status. Interestingly, individual fish generally retained their rank in swimming speed across different segments, except when transiting a challenging canyon midway during the migration. PMID- 18419522 TI - Hexamethylbisacetamide and disruption of human immunodeficiency virus type 1 latency in CD4(+) T cells. AB - BACKGROUND: Novel therapeutic approaches are needed to attack persistent proviral human immunodeficiency type 1 (HIV-1) infection. Hexamethylbisacetamide (HMBA), a hybrid bipolar compound, induces expression of the HIV-1 promoter in the long terminal repeat (LTR) region in a Tat-independent manner but mimics the effect of Tat, overcoming barriers to LTR expression and increasing the processivity of LTR transcription complexes. METHODS: We studied alterations in cellular factors and their LTR occupancy induced by HMBA in models of latent HIV-1 infection. We measured the induction of viral outgrowth by HMBA in resting CD4(+) T cells from aviremic HIV-1-infected donors. RESULTS: HMBA induced outgrowth of HIV-1 from resting CD4(+) T cells recovered from aviremic patients treated with antiretroviral therapy (ART). HMBA triggered cyclin-dependent kinase 9 (CDK9) recruitment to the LTR, a key factor in the induction of efficient HIV-1 expression, via an unexpected interaction with the transcription factor Sp1. The availability of Sp1 and Sp1 DNA binding sites were necessary for HMBA-induced CDK9 recruitment and LTR expression. HMBA signaling via both protein kinase C mu and phosphatidylinositol 3-kinase appeared to contribute to LTR induction. CONCLUSIONS: The novel mechanism through which HMBA disrupts latent HIV-1 infection involves 2 cellular kinases that may be therapeutically exploited to induce expression of persistent proviral HIV-1. PMID- 18419523 TI - An empirical test of a diffusion model: predicting clouded apollo movements in a novel environment. AB - Functional connectivity is a fundamental concept in conservation biology because it sets the level of migration and gene flow among local populations. However, functional connectivity is difficult to measure, largely because it is hard to acquire and analyze movement data from heterogeneous landscapes. Here we apply a Bayesian state-space framework to parameterize a diffusion-based movement model using capture-recapture data on the endangered clouded apollo butterfly. We test whether the model is able to disentangle the inherent movement behavior of the species from landscape structure and sampling artifacts, which is a necessity if the model is to be used to examine how movements depend on landscape structure. We show that this is the case by demonstrating that the model, parameterized with data from a reference landscape, correctly predicts movements in a structurally different landscape. In particular, the model helps to explain why a movement corridor that was constructed as a management measure failed to increase movement among local populations. We illustrate how the parameterized model can be used to derive biologically relevant measures of functional connectivity, thus linking movement data with models of spatial population dynamics. PMID- 18419524 TI - The tempo and mode of three-dimensional morphological evolution in male reproductive structures. AB - Various evolutionary forces may shape the evolution of traits that influence the mating decisions of males and females. Phenotypic traits that males and females use to judge the species identify of potential mates should evolve in a punctuated fashion, changing significantly at the time of speciation but changing little between speciation events. In contrast, traits experiencing sexual selection or sexually antagonistic interactions are generally expected to change continuously over time because of the directional selection pressures imposed on one sex by the actions of the other. To test these hypotheses, we used spherical harmonic representations of the shapes of male mating structures in reconstructions of the evolutionary tempo of these structures across the history of the Enallagma damselfly clade. Our analyses show that the evolution of these structures is completely consistent with a punctuated model of evolutionary change and a constant evolutionary rate throughout the clade's history. In addition, no interpopulation variation in shape was detected across the range of one species. These results indicate that male mating structures in this genus are used primarily for identifying the species of potential mates and experience little or no selection from intraspecific sexual selection or sexual antagonism. The implications of these results for speciation are discussed. PMID- 18419525 TI - Federal funding for the study of antimicrobial resistance in nosocomial pathogens: no ESKAPE. PMID- 18419526 TI - It's time for a change in practice: reducing antibiotic use can alter antibiotic resistance. PMID- 18419527 TI - The research agenda of the National Institute of Allergy and Infectious Diseases for antimicrobial resistance. AB - Antimicrobial resistance is an intrinsic and inevitable aspect of microbial survival that continually challenges human health. Research on antimicrobial resistance is central to the mission of the National Institute of Allergy and Infectious Diseases (NIAID). In fiscal year 2007, NIAID invested more than USD800 million to support basic and translational research on antimicrobials, more than USD200 million of which is devoted to understanding the causes, consequences, and treatments of antimicrobial drug resistance. The complex process that facilitates the transformation of ideas into therapies requires a pipeline that runs from bench to bedside, and NIAID has leveraged the entire spectrum of conventional and biodefense resources. NIAID works in partnership with other federal agencies, industry, foundation partners, and foreign governments. The basic and clinical research supported by NIAID will, ideally, continue to yield profound rewards in terms of the understanding, diagnosis, and treatment of infectious diseases. PMID- 18419528 TI - Seasonality of antibiotic-resistant streptococcus pneumoniae that causes acute otitis media: a clue for an antibiotic-restriction policy? AB - BACKGROUND: It is unclear whether reducing antibiotic prescriptions can reduce rates of resistance once resistance becomes prevalent. We attempted to determine whether reduced antibiotic consumption, which is observed yearly in children during the warm season, is associated with a reduction in antibiotic resistance in pneumococcal acute otitis media (AOM). METHODS: Antibiotic prescriptions and resistance were measured prospectively during 1999-2003 in 2 demographically distinct populations: Jewish and Bedouin children (aged <5 years) in southern Israel. Associations were assessed using seasonally clustered logistic regression models. RESULTS: The study included 236,466 prescriptions and 3609 pneumococcal isolates. Prescription rates decreased during the warm months by 36% and 15% in Jewish and Bedouin children, respectively (P < .001 for the season). Among Jewish children, higher resistance rates were observed during the cold than the warm months (P < .001 for each antibiotic). This difference remained significant after adjustment for age, ethnic group, study year, history of antibiotic use, and serotype. The difference was not observed in Bedouin children. CONCLUSIONS: Rapid seasonal decline in resistant AOM-causing pneumococci occurred only in Jewish children, among whom a marked prescribing seasonality was noted, and not in Bedouin children, among whom prescription was less seasonal. The rapid seasonal decrease in resistance associated with markedly reduced antibiotic use suggests that drug-resistant pneumococci may pay a fitness cost. PMID- 18419530 TI - Second dose of varicella vaccine for children: are we giving it too late? PMID- 18419529 TI - Reovirus preferentially infects the basolateral surface and is released from the apical surface of polarized human respiratory epithelial cells. AB - Mammalian reoviruses infect respiratory and gastrointestinal epithelia and cause disease in neonates. Junctional adhesion molecule-A (JAM-A) is a serotype independent receptor for reovirus. JAM-A localizes to tight junctions and contributes to paracellular permeability in polarized epithelia. To investigate the mechanisms of reovirus infection of polarized epithelial cells, we assessed reovirus replication, release, and spread after apical and basolateral adsorption to primary human airway epithelial cultures. Reovirus infection of human airway epithelia was more efficient after adsorption to the basolateral surface than after adsorption to the apical surface, and it was dependent on JAM-A. Reovirus binding to carbohydrate coreceptor sialic acid inhibited apical infection, which was partially ameliorated by treatment of the cultures with neuraminidase. Despite the preference for basolateral infection, reovirus was released from the apical surface of respiratory epithelia and did not disrupt tight junctions. These results establish the existence of an infectious circuit for reovirus in polarized human respiratory epithelial cells. PMID- 18419531 TI - T Cells and Tuberculosis: Beyond Interferon-gamma. PMID- 18419532 TI - Primary vaccine failure after 1 dose of varicella vaccine in healthy children. AB - Universal immunization of young children with 1 dose of varicella vaccine was recommended in the United States in 1995, and it has significantly decreased the incidence of chickenpox. Outbreaks of varicella, however, are reported among vaccinated children. Although vaccine effectiveness has usually been 85%, rates as low as 44% have been observed. Whether this is from primary or secondary vaccine failure-or both-is unclear. We tested serum samples from 148 healthy children immunized against varicella in New York, Tennessee, and California to determine their seroconversion rates, before and after 1 dose of Merck/Oka varicella vaccine. The median age at vaccination was 12.5 months; postvaccination serum samples were obtained on average 4 months later. Serum was tested for antibodies against varicella-zoster virus (VZV) by use of the previously validated sensitive and specific fluorescent antibody to membrane antigen (FAMA) assay. Of 148 healthy child vaccinees, 113 (76%) seroconverted, and 24% had no detectable VZV FAMA antibodies. Our data contrast with reported seroconversion rates of 86%-96% by other VZV antibody tests and suggest that many cases of varicella in immunized children are due to primary vaccine failure. A second dose of varicella vaccine is expected to increase seroconversion rates and vaccine effectiveness. PMID- 18419533 TI - Colonization of mice by Candida albicans is promoted by chemically induced colitis and augments inflammatory responses through galectin-3. AB - BACKGROUND: Little is known about the relationship between colonic inflammation and Candida albicans colonization. Galectin-3 (Gal-3) is an intestinal lectin that binds to specific C. albicans glycans and is involved in inflammation. METHODS: Colitis was experimentally induced in wild-type and Gal3(-/-) mice using dextran sulfate sodium (DSS) before oral administration of C. albicans. Yeast recovered from stools was quantified. The presence of yeast and inflammation were evaluated in sections of colon by histologic examination, quantification of myeloperoxidase (MPO) activity, and by gene expression for cytokines and innate immune receptors. Serum from mice was collected for determination of anti-yeast mannan antibodies, including anti-Saccharomyces cerevisiae antibodies (ASCA), which are biomarkers of an inflammatory bowel disease. RESULTS: Inflammation strongly promoted C. albicans colonization. Conversely, C. albicans augmented inflammation induced by DSS, as assessed by histologic scores, MPO activity, and tumor necrosis factor (TNF)-alpha and Toll-like receptor (TLR)-2 expression. C. albicans colonization generated ASCA. The absence of Gal-3 reduced DSS inflammation and abolished the response of TLR-2 and TNF-alpha to C. albicans colonization. CONCLUSIONS: DSS-induced colitis provides a model for establishing C. albicans colonization in mice. This model reveals that C. albicans augments inflammation and confirms the role of Gal-3 in both inflammation and the control of host responses to C. albicans. PMID- 18419534 TI - Superantigen-like effects of a Candida albicans polypeptide. AB - The amino terminal sequence of the Candida albicans cell wall protein Int1 exhibited partial identity with the major histocompatibility complex (MHC) class II binding site of the Mycoplasma arthritidis superantigen MAM. Int1-positive C. albicans blastospores activated human T lymphocytes and expanded Vbeta subsets 2, 3, and/or 14; Int1-negative strains were inactive. Release of interferon-gamma (IFN-gamma) but not of tumor necrosis factor-alpha or interleukin-6 was Int1 dependent; interleukin-4 and interleukin-10 were not detected. T lymphocyte activation, Vbeta expansion, and IFN-gamma release were associated with a soluble polypeptide that encompassed the first 263 amino acids of Int1 (Pep(263)). Monoclonal antibody 163.5, which recognizes an Int1 epitope that overlaps the region of identity with MAM, significantly inhibited these activities when triggered by Int1-positive blastospores or Pep(263) but not by staphylococcal enterotoxin B. Histidine(263) was required. Pep(263) bound to T lymphocytes and MHC class II and was detected in the urine of a patient with C. albicans fungemia. These studies identify a candidal protein that displays superantigen like activities. PMID- 18419535 TI - Detection of polyfunctional Mycobacterium tuberculosis-specific T cells and association with viral load in HIV-1-infected persons. AB - BACKGROUND: The human immunodeficiency virus type 1 (HIV-1) epidemic is associated with a significant increase in the incidence of tuberculosis (TB); however, little is known about the quality of Mycobacterium tuberculosis (MTB) specific cellular immune responses in coinfected individuals. METHODS: A total of 137 HIV-1-positive individuals in Durban, South Africa, were screened with the use of overlapping peptides spanning Ag85A, culture filtrate protein 10 (CFP-10), early secretory antigen target 6 (ESAT-6), and TB10.4, in an interferon (IFN) gamma enzyme-linked immunospot (ELISPOT) assay. Intracellular cytokine staining for MTB-specific production of IFN-gamma, tumor necrosis factor (TNF)-alpha, and interleukin (IL)-2 was performed, as was ex vivo phenotyping of memory markers on MTB-specific T cells. RESULTS: A total of 41% of subjects responded to ESAT-6 and/or CFP-10, indicating the presence of latent MTB infection. The proportion of MTB-specific IFN-gamma(+)/TNF-alpha(+) CD4(+) cells was significantly higher than the proportion of IFN-gamma(+)/IL-2(+) CD4(+) cells (P = .0220), and the proportion of MTB-specific IL-2-secreting CD4 cells was inversely correlated with the HIV-1 load (P = .0098). MTB-specific CD8 T cells were predominately IFN gamma(+)/TNF-alpha(+)/IL-2(-). Ex vivo memory phenotyping of MTB-specific CD4 and CD8 T cells indicated an early to intermediate differentiated phenotype for the population of effector memory cells. CONCLUSIONS: Polyfunctional MTB-specific CD4 and CD8 T cell responses are maintained in the peripheral blood of HIV-1-positive individuals, in the absence of active disease, and the functional capacity of these responses is affected by HIV-1 disease status. PMID- 18419536 TI - Impact of human immunodeficiency virus infection on Streptococcus pneumoniae colonization and seroepidemiology among Zambian women. AB - Nasopharyngeal colonization with Streptococcus pneumoniae precedes invasive pneumococcal disease. Human immunodeficiency virus (HIV) infection increases rates of invasive pneumococcal disease, and its effect on colonization is unknown. In a longitudinal cohort of Zambian mothers with or without HIV infection, HIV infection increased the risk of colonization (risk ratio [RR], 1.9; 95% confidence interval [CI], 1.3-2.8) and repeat colonization (RR, 2.4; 95% CI, 1.1-5.3) and reduced the time to new colonization (P = .01). Repeat colonization with homologous sero/factor types occurred only among HIV-positive mothers. Pediatric serotypes 6, 19, and 23 accounted for excess colonization among HIV-positive mothers. HIV infection significantly increases the risk of pneumococcal colonization. Increased rates of colonization by pediatric serotypes suggest a potential role for the 7-valent pneumococcal vaccine in HIV-infected adults. PMID- 18419537 TI - Efavirenz-based regimens in treatment-naive patients with a range of pretreatment HIV-1 RNA levels and CD4 cell counts. AB - The potency of 2 nucleoside reverse transcriptase inhibitors (NRTIs) and efavirenz in patients with higher viral loads (VLs) or low CD4 cell counts remains uncertain. Virologic failure and changes in CD4 count in relation to pretreatment VL and CD4 count were evaluated in treatment-naive patients randomized to treatment groups that received 2 or 3 NRTIs with efavirenz. Over 3 years, the risk of virologic failure was not significantly different among subgroups categorized according to pretreatment VL or CD4 count. No significant differences among subgroups were observed for CD4 count changes, except in patients with high pretreatment VL. There were no significant differences among subgroups with respect to treatment responses. These results demonstrate the potency of efavirenz-containing regimens across a spectrum of pretreatment VLs and CD4 counts. PMID- 18419538 TI - HIV-1 variation before seroconversion in men who have sex with men: analysis of acute/early HIV infection in the multicenter AIDS cohort study. AB - Understanding the characteristics of human immunodeficiency virus (HIV) necessary for infection in a new host is a critical goal for acquired immunodeficiency syndrome (AIDS) research. We studied the characteristics of HIV-1 envelope genes in 38 men in the Multicenter AIDS Cohort Study cohort before seroconversion. We found a range of diversity (0.2%-5.6% [median, 0.86%]), V1-V2 loop length (58-93 aa), and potential N-linked glycosylation sites (n = 2-9). However, at least 46% of the men had replicating virus that appeared to have been derived from a single viral variant. Nearly all variants were predicted to be CCR5 tropic. We found no correlation between these viral characteristics and the HIV outcomes of time to clinical AIDS or death and/or a CD4 cell count <200 cells/microL. PMID- 18419539 TI - Population snapshot of emergent Streptococcus pneumoniae serotype 19A in the United States, 2005. AB - BACKGROUND: Serotype 19A invasive pneumococcal disease (IPD) increased annually in the United States after the introduction of the 7-valent conjugate vaccine (PCV7). To understand this increase, we characterized serotype 19A isolates recovered during 2005. METHODS: IPD cases during 1998-2005 were identified through population-based surveillance. We performed susceptibility testing and multilocus sequence typing on 528 (95%) of 554 serotype 19A isolates reported in 2005. RESULTS: The incidence of IPD due to serotype 19A increased from 0.8 to 2.5 cases per 100,000 population between 1998 and 2005 (P < .05), whereas the overall incidence of IPD decreased from 24.4 to 13.8 cases per 100,000 population (P < .05). Simultaneously, the incidence of IPD due to penicillin-resistant 19A isolates increased from 6.7% to 35% (P < .0001). Of 151 penicillin-resistant 19A isolates, 111 (73.5%) belonged to the rapidly emerging clonal complex 320, which is related to multidrug-resistant Taiwan(19F)-14. The remaining penicillin resistant strains were highly related to other clones of PCV7 serotypes or to isolates within major 19A clonal complex 199 (CC199). In 1999, only CC199 and 3 minor clones were apparent among serotype 19A isolates. During 2005, 11 multiple isolate clonal sets were detected, including capsular switch variants of a serotype 4 clone. CONCLUSIONS: PCV7 ineffectiveness against serotype 19A, antibiotic resistance, clonal expansion and emergence, and capsular switching have contributed to the genetic diversity of 19A and to its emergence as the predominant invasive pneumococcal serotype in the United States. PMID- 18419540 TI - Biofilm formation induces C3a release and protects Staphylococcus epidermidis from IgG and complement deposition and from neutrophil-dependent killing. AB - BACKGROUND: Biofilm formation is considered to be an important virulence factor of the opportunistic pathogen Staphylococcus epidermidis. We hypothesized that biofilm formation could interfere with the deposition of immunoglobulins and complement on the bacterial surface, leading to diminished activation of the complement system and protection from killing by human phagocytes. METHODS: The killing of biofilm-encased and planktonically grown wild-type (wt) S. epidermidis and the killing of an isogenic biofilm-negative ica mutant (ica(-)) by human polymorphonuclear neutrophils (PMNs) were compared. C3a induction and deposition of C3b and immunoglobulin G (IgG) on the bacteria after opsonization with human serum were assessed by enzyme-linked immunosorbent assay, flow cytometry, and electron microscopy. The virulence of the bacterial strains was compared in a mouse model of catheter-associated infection. RESULTS: Biofilm-embedded wt S. epidermidis was killed less well by human PMNs and induced more C3a than planktonically grown wt and ica(-) S. epidermidis. However, the deposition of C3b and IgG on the bacterial surface was diminished in biofilm-encased staphylococci. wt S. epidermidis was more virulent in implant-associated infections and was killed more slowly than ica(-) in ex vivo assays of killing by PMNs. CONCLUSIONS: The results indicate that prevention of C3b and IgG deposition on the bacterial surface contributes to the biofilm-mediated protection of S. epidermidis from killing by PMNs. PMID- 18419541 TI - Staphylococcus aureus enterotoxin B regulates prostaglandin E2 synthesis, growth, and migration in nasal tissue fibroblasts. AB - BACKGROUND: Superantigens and eicosanoids are important amplifiers and regulators of inflammation in airway diseases. We therefore studied the possible influence of Staphylococcus aureus enterotoxin B (SEB) on the cyclooxygenase (COX) pathway and basic functions of airway structural cells. METHODS: Fibroblasts were isolated from nasal inferior turbinate tissue and cultured in the presence of different concentrations of SEB. Preincubation with interferon (IFN)-gamma was performed to induce expression of major histocompatibility complex (MHC) class II receptors. Prostaglandin E2 (PGE(2)) production was assayed by enzyme-linked immunosorbent assay, and levels of COX-2 and prostanoid E receptors 1-4 (EP(1-4)) were assayed by real-time polymerase chain reaction. Migration and growth tests were performed, and SEB was localized within the cells by confocal microscopy. RESULTS: Stimulation with IFN-gamma and SEB significantly down-regulated PGE(2), COX-2, and EP(2) expression but not EP(1), EP(3), or EP(4) expression. The enterotoxin blocked cell growth but increased the fibroblast migration rate. SEB was localized within the cell in the presence and absence of MHC-II, suggesting that mechanisms other than conventional binding may allow the enterotoxin to enter the cell. CONCLUSIONS: These findings may have major implications for our understanding of the role played by bacterial superantigens in regulating the inflammatory and remodeling mechanisms of upper airway diseases and hence may help elucidate the pathophysiology of these diseases. PMID- 18419542 TI - Complement-dependent synergistic bactericidal activity of antibodies against factor H-binding protein, a sparsely distributed meningococcal vaccine antigen. AB - BACKGROUND: Antibodies to factor H (fH)-binding protein (fHBP), a meningococcal vaccine antigen, activate classical complement pathway serum bactericidal activity (SBA) and block binding of the complement inhibitor fH. METHODS: To understand these 2 functions in protection, we investigated the interactions of human complement and 2 anti-fHBP monoclonal antibodies (MAbs) with encapsulated Neisseria meningitidis. RESULTS: JAR 3 (IgG3) blocks fH binding and elicits SBA against 2 strains with naturally high fHBP expression and a low-expressing strain genetically engineered to express high fHBP levels. JAR 4 (IgG2a) does not block fH binding or elicit SBA. Neither MAb alone elicits SBA against 2 other strains with low fHBP expression, but together the MAbs increase C4b binding and elicit SBA; JAR 3 alone also is bactericidal in whole blood. In nonimmune blood, fHBP knockout mutants from high-expressing stains do not survive, but mutants of low expressing strains do. CONCLUSIONS: Expression of fHBP is a prerequisite for bacterial survival in blood only by strains with naturally high fHBP expression. In low-expressing strains, combinations of 2 nonbactericidal anti-fHBP MAbs can bind to nonoverlapping epitopes, engage C1q, activate C4, and mediate classical complement pathway SBA. In the absence of sufficient C4b binding for SBA, an individual MAb can have opsonophagocytic bactericidal activity. PMID- 18419543 TI - Pregnancy and HIV disease progression: methodological concerns. PMID- 18419544 TI - Effect of pregnancy on HIV disease progression during the era of highly active antiretroviral therapy. PMID- 18419546 TI - Interleukin-8 induction by Burkholderia pseudomallei can occur without Toll-like receptor signaling but requires a functional type III secretion system. AB - Burkholderia pseudomallei is the causative agent of melioidosis, an infectious disease that can result in asymptomatic, chronic, or acute illness. In acute melioidosis, high levels of proinflammatory cytokines and chemokines are found in organs and blood, and neutrophils play a key role in controlling the infection. We showed that B. pseudomallei activates NF-kappaB via Toll-like receptor (TLR) 2, TLR4, and TLR5 but can also activate NF-kappaB and induce interleukin (IL)-8 without involving TLRs. TLR-independent activation depends on a functional Bsa type III secretion system (T3SS) and requires internalization. The mitogen activated protein kinase (MAPK) inhibitors for p38 and c-Jun N-terminal kinase (JNK) severely impaired IL-8 induction by B. pseudomallei and reduced bacterial internalization. Furthermore, the T3SS mutant induced less JNK phosphorylation than did wild-type bacteria. Thus, in cells with no or low expression of TLRs, such as mucosal epithelial cells, B. pseudomallei can induce IL-8 via NF-kappaB and MAPK pathways, aided by Bsa T3SS. PMID- 18419547 TI - Type-specific duration of human papillomavirus infection: implications for human papillomavirus screening and vaccination. AB - BACKGROUND: Understanding the duration of human papillomavirus (HPV) infection may help find suitable end points for vaccine trials and testing intervals in screening studies. We studied genotype-specific infection duration among 2462 women enrolled in the Ludwig-McGill cohort study. METHODS: Cervical specimens collected every 4-6 months were tested by a polymerase chain reaction protocol. Actuarial techniques were used to estimate the duration of HPV infection and to investigate the influence of age, number of sexual partners, and coinfection with multiple HPV types. RESULTS: At enrollment, the prevalence of infection with high risk HPV types was 10.6%, and the prevalence of infection with low-risk HPV types was 6.1%; incidence rates were 6.1 and 5.0 infections per 1000 women-months, respectively. Prevalent infections took longer to clear than incident infections (mean time to clearance, 18.6 months vs. 13.5 months). The mean duration of incident infection with high- and low-risk HPV varied according to the analytic approach used to measure this variable and showed considerable variation by HPV type (range, 5.1-15.4 months). Age and number of partners did not influence infection duration, whereas coinfection was associated with increased infection duration. The mean duration of HPV-16 monoinfection was 11.0 months, and the mean duration of HPV-16 coinfection was 15.4 months. CONCLUSION: There was considerable variation among HPV types with regard to the duration of infection. Coinfection with multiple types contributed to an increased infection duration. PMID- 18419548 TI - Immune response during adverse events after 17D-derived yellow fever vaccination in Europe. AB - BACKGROUND: In 1999-2000, reports of fatalities after vaccination with 17D derived yellow fever vaccine (YEL) focused mainly on cases of YEL-associated adverse events (YEL-AEs) and YEL-associated viscerotropic disease (YEL-AVD). Here, we investigated 6 recent European cases to provide insight regarding immune response involvement and to identify potential risk factors. METHODS: Clinical, microbiological, molecular biological, and immunological assays were performed on serum from 6 patients with YEL-AEs, including 5 with YEL-AVD and 1 with YEL associated neurotropic disease (YEL-AND). RESULTS: The levels of 3 liver enzymes associated with infection were clearly increased in all patients with YEL-AVD, but no elevations were observed in the patient with YEL-AND. In the patients with severe YEL-AVD, platelet counts were markedly reduced (< 100,000 cells/microL). The only patient with fatal YEL-AVD exhibited a cytokine profile comparable to that seen in YF: high levels of interleukin (IL)-6, IL-8, monocyte chemotactic protein (MCP)-1, monokine induced by interferon-gamma, and growth-related oncogene (GRO). The other patients with YEL-AVD exhibited similar but less severe cytokine profiles. The patient with YEL-AND exhibited a cytokine profile similar to that found in vaccinees without YEL-AEs: elevated levels of RANTES and low levels of GRO, MCP-1, transforming growth factor-beta1, and tumor necrosis factor beta. CONCLUSIONS: On the basis of these results, we conclude that elevations in cytokine levels and reductions in platelet counts are suitable surrogate markers for patients likely to experience severe adverse reactions to YEL. PMID- 18419549 TI - Specific enfuvirtide-associated mutational pathways in HIV-1 Gp41 are significantly correlated with an increase in CD4(+) cell count, despite virological failure. AB - BACKGROUND: Human immunodeficiency virus type 1 (HIV-1) gp41 is a crucial determinant for HIV-1 pathogenicity. We investigated the correlation of enfuvirtide (ENF)-associated gp41 mutational clusters with viroimmunological parameters, as well as the potential underlying mechanisms. METHODS: A total of 172 gp41 sequences and clinical follow-up data from 73 ENF-treated patients were analyzed monthly, from baseline to week 48. RESULTS: There were 7 novel gp41 mutations positively associated with ENF treatment and correlated with classic ENF mutations. The ENF-associated clusters [V38A + N140I ] and [V 38A +T18A ] significantly correlated with an increase in CD4 cell count at week 48 ( an increase from baseline of 112 and 209 cells/microL, respectively), whereas [Q40H + L45M + 268A] significantly correlated with a decrease in CD4 cell count (-53 cells/microL), without a change in the level of viremia. Residues 38 and 18 are located complementarily to each other in the Rev-responsive element, whereas analysis of molecular dynamics showed that the copresence of [V38A + N140 I] abolishes the interaction between residue 38 and 145 important for stabilization of the 6-helix bundle. In contrast, T268A localizes in the gp41 calmodulin binding domain responsible for gp41-induced CD4(+) T lymphocyte apoptosis. CONCLUSION: Specific gp41 mutational clusters associated with ENF treatment significantly correlate with increases in CD4(+) cell count. Structural analysis suggests that this immunological gain is associated with mechanisms that act at both the protein level and the RNA level (even under conditions of virological failure). This result may help in the selection of patients who can benefit most from ENF treatment and represents a driving force for the design of the next generation of entry inhibitors. PMID- 18419551 TI - Plant genotype shapes ant-aphid interactions: implications for community structure and indirect plant defense. AB - Little is known about the mechanisms by which plant genotype shapes arthropod community structure. In a field experiment, we measured the effects of milkweed (Asclepias syriaca) genotype and ants on milkweed arthropods. Populations of the ant-tended aphid Aphis asclepiadis and the untended aphid Myzocallis asclepiadis varied eight- to 18-fold among milkweed genotypes, depending on aphid species and whether ants were present. There was no milkweed effect on predatory arthropods. Ants increased Aphis abundance 59%, decreased Myzocallis abundance 52%, and decreased predator abundance 56%. Milkweed genotype indirectly influenced ants via direct effects on Aphis and Myzocallis abundance. Milkweed genotype also modified ant-aphid interactions, influencing the number of ants attracted per Aphis and Myzocallis. While ant effects on Myzocallis were consistently negative, effects on Aphis ranged from antagonistic to mutualistic among milkweed genotypes. As a consequence of milkweed effects on ant-aphid interactions, ant abundance varied 13-fold among milkweed genotypes, and monarch caterpillar survival was negatively correlated with genetic variation in ant abundance. We speculate that heritable variation in milkweed phloem sap drives these effects on aphids, ants, and caterpillars. In summary, milkweed exerts genetic control over the interactions between aphids and an ant that provides defense against foliage feeding caterpillars. PMID- 18419550 TI - Cognitive dysfunction in mice infected with Plasmodium berghei strain ANKA. AB - Cerebral malaria complicated by cognitive sequelae is a major cause of morbidity in humans infected with Plasmodium falciparum. To model cognitive function after malaria, we created a rodent model of cerebral malaria by infecting C57BL/6 mice with Plasmodium berghei strain ANKA. After 7 days, an object-recognition test of working memory revealed a significant impairment in the visual memory of infected mice. This impairment was observed in the absence of confounding effects of infection. The cognitive dysfunction correlated with hemorrhage and inflammation. Furthermore, microglial activity and morphological changes detected throughout the brains of infected mice were absent from the brains of control mice, and this correlated with the measured cognitive defects. Similar testing methods in human studies could help identify subjects at risk for an adverse cognitive outcome. This murine model should facilitate the study of adjunctive methods to ameliorate adverse neurological outcomes in cerebral malaria. PMID- 18419552 TI - The energetic advantages of slug specialization in garter snakes (genus Thamnophis). AB - We tested the hypothesis that dietary specialization by foraging garter snakes is accompanied by increased assimilation efficiency on specialist prey items. Our comparison included two closely related garter snake species considered to be slug specialists (Thamnophis ordinoides and Thamnophis elegans terrestris), one fish specialist (Thamnophis couchii), and one diet generalist (Thamnophis elegans elegans). Our results suggest that slug specialists have an energetic advantage over non-slug-eating snakes when both eat slugs. Slug specialists T. ordinoides and T. e. terrestris both have higher assimilation and net assimilation efficiencies when eating slugs than do generalists T. e. elegans and T. couchii. The slug specialists did not experience decreased efficiency when eating fish. Therefore, there was no apparent digestive trade-off for the slug specialists when eating other prey. PMID- 18419553 TI - Body condition at weaning affects the duration of the postweaning fast in gray seal pups (Halichoerus grypus). AB - Gray seals (Halichoerus grypus) undergo a terrestrial postweaning fast (PWF) that depletes energy reserves acquired during the suckling interval. Plasticity in PWF duration may ensure that pups of variable body condition depart for sea with adequate energy reserves. To test this hypothesis, we examined body condition of 30 gray seal pups at weaning and monitored their PWF duration. On average, fat accounted for 47.3% +/- 0.7% of their 53.2 +/- 1.3-kg weaning mass. Although fasting duration averaged 21 +/- 1.1 d (n = p28), there was considerable variation in fasting duration (9 to > 31 d) and the resulting age when pups departed to sea (26 to > 49 d). Percent fat at weaning(38.6%-54.6%) was positively correlated with fasting duration(n = 28, r = 0.376, P = 0.0489). In contrast, total body gross energy (735.3-1,447.4 MJ) and body mass (39.0-66.0 kg) were not correlated with fasting duration. Thus, body composition,not overall body reserves, predicted fasting duration, but the effect was weak, indicating that other factors also account for the observed variation in fasting duration. We speculate that pups with greater percent fat more effectively utilized lipid and conserved protein while meeting metabolic costs throughout the PWF. As a result, fatter pups extended the PWF duration,which may be critical for development of diving physiology and may have facilitated their survivorship to age 1. PMID- 18419554 TI - Is the alkaline tide a signal to activate metabolic or ionoregulatory enzymes in the dogfish shark (Squalus acanthias)? AB - Experimental metabolic alkalosis is known to stimulate whole-animal urea production and active ion secretion by the rectal gland in the dogfish shark. Furthermore, recent evidence indicates that a marked alkaline tide (systemic metabolic alkalosis) follows feeding in this species and that the activities of the enzymes of the ornithine-urea cycle (OUC) for urea synthesis in skeletal muscle and liver and of energy metabolism and ion transport in the rectal gland are increased at this time. We therefore evaluated whether alkalosis and/or NaCl/volume loading (which also occurs with feeding) could serve as a signal for activation of these enzymes independent of nutrient loading. Fasted dogfish were infused for 20 h with either 500 mmol L(-1) NaHCO3 (alkalosis + volume expansion) or 500 mmol L(-1) NaCl (volume expansion alone), both isosmotic to dogfish plasma, at a rate of 3 mL kg(-1) h(-1). NaHCO3 infusion progressively raised arterial pH to 8.28 (control = 7.85) and plasma [HCO3-] to 20.8 mmol L(-1) (control = 4.5 mmol L(-1)) at 20 h, with unchanged arterial P(CO2), whereas NaCl/volume loading had no effect on blood acid-base status. Rectal gland Na+,K+ ATPase activity was increased 50% by NaCl loading and more than 100% by NaHCO3 loading, indicating stimulatory effects of both volume expansion and alkalosis. Rectal gland lactate dehydrogenase activity was elevated 25% by both treatments, indicating volume expansion effects only, whereas neither treatment increased the activities of the aerobic enzymes citrate synthase, NADP-isocitrate dehydrogenase, or the ketone body-utilizing enzyme beta-hydroxybutyrate dehydrogenase in the rectal gland or liver. The activity of ornithine-citrulline transcarbamoylase in skeletal muscle was doubled by NaHCO3 infusion, but neither treatment altered the activities of other OUC-related enzymes (glutamine synthetase, carbamoylphosphate synthetase III). We conclude that both the alkaline tide and salt loading/volume expansion act as signals to activate some but not all of the elevated metabolic pathways and ionoregulatory mechanisms needed during processing of a meal. PMID- 18419555 TI - Basal metabolic rate of aged mice is affected by random genetic drift but not by selective breeding for high early-age locomotor activity or chronic wheel access. AB - The study of correlated evolution can lead to new insights about the inheritance patterns of complex traits. In order to better understand the evolution of metabolic rate, we tested whether voluntary activity levels and basal metabolic rate are genetically correlated in 90-wk-old mice (Mus domesticus) from replicated lines of the sixteenth generation of an artificial selection experiment for high early-age wheel-running activity. We measured basal rates of oxygen consumption and carbon dioxide production and also computed the respiratory exchange ratio. Half of the individuals from both selected and control lines had been allowed free access to running wheels since 4 wk of age, while the other half were in standard cages. This design allowed testing of hypotheses about (1) genetic correlations between voluntary activity and metabolic rate and (2) lifetime training effects on metabolic traits. Selection group did not have a significant effect on metabolic traits; therefore, this study does not support some of the implicit assumptions of the aerobic capacity model for the evolution of vertebrate energetics. Activity group also did not affect metabolic rate, indicating that lifetime training does not alter basal metabolism in these mice. However, strong replicate line-within-selection-group differences were detected, indicating the occurrence of random genetic drift. In females, this divergence in metabolic traits attributable to drift was independent of body mass, but in males it was probably caused by a correlated response to selection involving body mass. This study is the first to show such effects of random genetic drift on metabolic traits. PMID- 18419556 TI - Thermal plasticity of diving behavior, aquatic respiration, and locomotor performance in the Mary River turtle Elusor macrurus. AB - Locomotion is a common measure of performance used in studies of thermal acclimation because of its correlation with predator escape and prey capture. However, for sedentary animals such as freshwater turtles, we propose that diving behavior may be a more ecologically relevant measure of performance. Increasing dive duration in hatchling turtles reduces predator exposure and therefore functions as an ecological benefit. Diving behavior is thermally dependent, and in some species of freshwater turtles, it is also reliant on aquatic respiration. This study examined the influence of thermal acclimation on diving behavior, aquatic respiration, and locomotor performance in the endangered, bimodally respiring Mary River turtle Elusor macrurus. Diving behavior was found to partially acclimate at 17 degrees C, with turtles acclimated to a cold temperature (17 degrees C) having a significantly longer dive duration than hatchlings acclimated to a warm temperature (28 degrees C). This increase in dive duration at 17 degrees C was not a result of physiological alterations in metabolic rate but was due instead to an increase in aquatic oxygen consumption. Increasing aquatic oxygen consumption permitted cold-acclimated hatchlings to remain submerged for significantly longer periods, with one turtle undertaking a dive of over 2.5 d. When burst-swimming speed was used as the measure of performance, thermal acclimation was not detected. Overall, E. macrurus demonstrated a partial ability to acclimate to changes in environmental temperature. PMID- 18419557 TI - The osmorespiratory compromise in sculpins: impaired gas exchange is associated with freshwater tolerance. AB - We acclimated two species of sculpin, the freshwater prickly sculpin (Cottus asper) and the closely related marine Pacific staghorn sculpin (Leptocottus armatus) to freshwater ( approximately 0 g/L), brackish water (15 g/L), and seawater (30 g/L) for at least 4 wk and examined the relationships between respiration, ion regulation, gill morphology, and freshwater tolerance. The prickly sculpin successfully acclimated to all three salinities and did not experience appreciable changes in plasma osmolality, [Cl-], or mortality. Gill Na+/K+-ATPase activity was lowest in prickly sculpins acclimated to freshwater, their native salinity, and increased during acclimation to seawater. Furthermore, prickly sculpins acclimated to freshwater had a 30% higher P(crit) than fish acclimated to brackish water or seawater; P(crit) is the environmental P(O2) below which an animal can no longer maintain a routine (.-)M(O2), and an increase in P(crit) represents a compromise of respiratory gas exchange. The higher P(crit) observed in prickly sculpins acclimated to freshwater is likely a consequence of their having small, relatively thick gills that increase in thickness (by approximately 1 microm) during freshwater exposure. In contrast, the marine Pacific staghorn sculpin successfully acclimated to brackish water and seawater, but high mortality (25%) was observed after 3 wk of exposure to freshwater. Pacific staghorn sculpins exposed to freshwater suffered significant, 15%-20%, reductions in plasma osmolality and [Cl-], and these losses in plasma ions resulted in a 1.4-fold increase in gill Na+/K+-ATPase activity. Pacific staghorn sculpins have large, thin gills that are not modified in response to salinity acclimation, and as a result, these animals show no respiratory compromise during freshwater acclimation, as evidenced by the lack of change in P(crit), but show significant ion regulatory disturbance. Overall, this study suggests that gill thickening and the resulting respiratory compromise are necessary for freshwater tolerance in sculpins. PMID- 18419558 TI - Power isn't everything: muscle function and energetic costs during steady swimming in Atlantic cod (Gadus morhua). AB - Power produced by red myotomal muscles of fish during cruise swimming appears seldom maximized, so we sought to investigate whether economy may impact or dominate muscle function. We measured cost of transport (COT) using oxygen consumption and the strain trajectories and electromyographic activity of red muscle measured at anterior (ANT) and posterior (POST) locations while Atlantic cod (Gadus morhua) swam steadily at speeds between 0.3 and 1.0 body lengths (BL) s(-1). We then measured the power produced by isolated segments of red muscle when activated either as in the swimming cod or such that maximal net power was produced. Patterns of activation during swimming were not optimal for power output and were highly variable between tail beats, particularly at the ANT location and at slow swim speeds. Muscle strain amplitude did not increase until swimming speed reached 0.9 (ANT) versus 0.5 (POST) BL s(-1). These limited power to only 53% (ANT) and 71% (POST) of maximum at slower swim speeds and to 70%-80% of maximum at high swim speeds. COT (resting metabolism subtracted) was minimal at the slowest swim speed, surprisingly, where power was most impaired by activation and strain. Thus, production of powered forces for maneuverability/stability appeared to greatly impact red muscle function during cruise swimming in cod, particularly at slow speeds and in ANT muscle. PMID- 18419559 TI - Varying hydric conditions during incubation influence egg water exchange and hatchling phenotype in the red-eared slider turtle. AB - Environmental conditions within the nest, notably temperature and moisture of substrate, exert a powerful influence during embryogenesis in oviparous reptiles. The influence of fluctuating nest temperatures has been experimentally examined in different reptile species; however, similar experiments using moisture as the key variable are lacking. In this article, we examine the effect of various substrate moisture regimes during incubation on different traits (egg mass, incubation length, and hatchling mass) in a chelonian species with flexible shelled eggs, the red-eared slider turtle (Trachemys scripta elegans). Our results show that the rate of water uptake by the eggs was higher in wet than in dry substrate and varied across development. More important, during the first third of development, the egg mass changes were relatively independent of the soil moisture level; they became very sensitive to moisture levels during the other two-thirds. Moreover, hydric conditions exerted a strong influence on the eggs' long-term sensitivity to the moisture of the substrate. Even short-term episodes of high or low levels of moisture modified permanently their water sensitivity, notably through modification of eggshell shape and volume, and in turn entailed significant effects on hatchling mass (and hence offspring quality). Such complex influences of fluctuating moisture levels at various incubation stages on hatchling phenotype better reflect the natural situation, compared to experiments based on stable, albeit different, moisture levels. PMID- 18419560 TI - Ovarian activity is differentially regulated across the breeding season in immature compared with adult American crows (Corvus brachyrhynchos). AB - American crow (Corvus brachyrhynchos) reproduction is seasonal. While photoperiod alters reproductive physiology and behavior in nesting adults, whether seasonal photoperiod changes alter ovarian activity in nonnesting adult and immature birds was not known. Ovarian follicular composition was examined in immature and nonnesting adult crows. Birds were divided into groups reflecting daylight hour exposure when captured: 12L, 12:30L, 13:30L, and 14L, representing early-, progressing-, peak-, and postbreeding seasons in southern California. Adult ovarian mass peaked in 13:30L and then decreased 65% by 14L. In contrast, immature ovarian mass peaked in 12L and decreased 58% by 14L. Whereas no change in adult follicle composition occurred, primordial follicle numbers increased 42%, and previtellogenic follicles increased 86% in immatures, resulting in an overall follicle number increase in immatures by 13:30L and 14L. Adult atretic follicles declined 68% by 14L, while immature atretic follicle numbers remained constant. Adult apoptotic transferase-mediated dUTP nick end labeling (TUNEL) positive follicle numbers remained constant until a 52% decrease occurred in 13:30L. TUNEL-positive follicles declined 58% by 12:30L among immatures. These data are the first to demonstrate that follicular composition is differentially regulated between nonnesting adult and immature American crows. Seasonal ovarian activity alterations in nonbreeding adults may serve to prime individuals for potential mating. PMID- 18419561 TI - Food restriction compromises immune memory in deer mice (Peromyscus maniculatus) by reducing spleen-derived antibody-producing B cell numbers. AB - Immune activity is variable in many wild animals, despite presumed strong selection against immune incompetence. Much variation may be due to changes in prevalence and abundance of pathogens (and/or their vectors) in time and space, but the costs of immune defenses themselves may also be important. Induction of immune activity often increases energy and protein expenditure, sometimes to the point of compromising fitness. Whether immune defenses are expensive to maintain once they are generated, however, is less well appreciated. If so, organisms would face persistent challenges of allocating resources between immunity and other expensive physiological processes, which would mandate trade-offs. Mild food restriction (70% ad lib. diet) reduces secondary antibody responses in deer mice (Peromyscus maniculatus), functionally representing a cost of immune memory. In this study, we asked whether compromised immune memory was mediated by a decrease in size of the B cell population responsible for producing antibodies (i.e., spleen-derived B lymphocytes producing immunoglobulin G [IgG]). Two weeks of food restriction reduced total splenocytes, total splenic B lymphocytes (B220+ cells), and splenic B lymphocytes producing IgG (B220+/IgG+ cells) but did not affect body mass or two circulating antibody subclasses (IgG1 and IgG2a) in deer mice. These results further indicate that maintenance of immune memory is expensive and may be subject to trade-offs with other physiological processes. PMID- 18419562 TI - Dual-energy X-ray absorptiometry (DXA) can accurately and nondestructively measure the body composition of small, free-living rodents. AB - Dual-energy x-ray absorptiometry (DXA) is a nondestructive technique that can potentially measure specific components of whole-body composition in free-living and lab-raised animals. Our aim was to test the ability of DXA to measure the composition of a common arvicoline rodent, the northern red-backed vole (Clethrionomys rutilus). We used a DXA apparatus to obtain measurements of fat mass (FM), lean mass (LM),bone mineral content, bone mineral density, and fat free mass(FFM) in carcasses of free-living and lab-raised voles. We then used chemical carcass analysis to derive predictive algorithms for actual values of FM, total body water, total protein, total mineral, LM, and FFM. Unexplained error in the equations for all voles grouped collectively ranged from R(2) = 0.82 to R(2) = 0.98. The DXA FM measurement had the highest coefficient of variation, and it was higher for free-living voles than for lab-raised voles. However, FM can be determined by difference with excellent precision by using the FFM equation (R(2) = 0.98). We also derived corrective terms for passive integrated transponder-tagged animals. Thus, DXA is a nonlethal, nondestructive tool capable of precisely and accurately measuring many specific parameters of whole-body composition in small free-living and lab-raised rodents. PMID- 18419564 TI - Moving beyond common-garden and transplant designs: insight into the causes of local adaptation in species interactions. AB - Theoretical and empirical studies of local adaptation in species interactions have increased greatly over the past decade, yielding new insights into the conditions that favor local adaptation or maladaptation. Generalizing the results of these studies is difficult, however, because of the different experimental designs that have been used to infer local adaptation. Particularly challenging is comparing results across empirical studies conducted in a common laboratory or garden environment with results of those conducted using transplants in natural environments. Here we develop simple and easily interpretable mathematical expressions for the quantities measured by these two different types of studies. Our results reveal that common-garden designs measure only a single component of local adaptation-the spatial covariance between the genotype frequencies of the interacting species-and thus provide only a partial description of local adaptation. In contrast, reciprocal-transplant designs incorporate additional terms that measure the contribution of spatial variability in the ecological environment. Consequently, the two types of studies should yield identical results only when local adaptation is caused by spatial variability in the genotype frequencies of the interacting species alone. In order to unify these disparate approaches, we develop a new methodology that can be used to estimate the individual components of local adaptation. When implemented in an appropriate experimental system, this partitioning allows the examination of fundamental questions such as the relative proportion of local adaptation attributable to interactions between species or to the abiotic environment. PMID- 18419565 TI - Spatially coupled larval supply of marine predators and their prey alters the predictions of metapopulation models. AB - Oceanographic forces can strongly affect the movement of planktonic marine larvae, often producing predictable spatial patterns of larval delivery. In particular, recent empirical evidence suggests that in some coastal systems, certain locations consistently receive higher (or lower) larval supplies of both predators and their prey. As a consequence, rates of settlement and predation may be coupled spatially, a phenomenon I term the "coupled settlement effect." To investigate the metapopulation consequences of this phenomenon, I created discrete-time, patch-based analytical and simulation models with a common larval pool and uneven larval supply among patches. Using two complementary measures of subpopulation value as a basis of comparison, I found that models with and without the coupled settlement effect yielded strikingly different predictions. When prey and predator larval supplies were not coupled, patches supplied with a larger proportion of the larval pool made a greater contribution to the metapopulation. When settlement of prey and predator was strongly coupled, however, the opposite was true: subpopulations with lower rates of larval supply (above some minimum) were more essential to metapopulation persistence. These considerations could facilitate more effective selection of sites for protection in marine reserves. PMID- 18419566 TI - Integrating across life-history stages: consequences of natal habitat effects on dispersal. AB - Ecological and evolutionary processes are affected by forces acting at both local and regional scales, yet our understanding of how these scales interact has remained limited. These processes are fundamentally linked through individuals that develop as juveniles in one environment and then either remain in the natal habitat or disperse to new environments. Empirical studies in a diverse range of organisms have demonstrated that the conditions experienced in the natal habitat can have profound effects on the adult phenotype. This environmentally induced phenotypic variation can in turn affect the probability that an individual will disperse to a new environment and the ecological and evolutionary impact of that individual in the new environment. We synthesize the literature on this process and propose a framework for exploring the linkage between local developmental environment and dispersal. We then discuss the ecological and evolutionary implications of dispersal asymmetries generated by the effects of natal habitat conditions on individual phenotypes. Our review indicates that the influence of natal habitat conditions on adult phenotypes may be a highly general mechanism affecting the flow of individuals between populations. The wealth of information already gathered on how local conditions affect adult phenotype can and should be integrated into the study of dispersal as a critical force in ecology and evolution. PMID- 18419567 TI - Trophically unique species are vulnerable to cascading extinction. AB - Understanding which species might become extinct and the consequences of such loss is critical. One consequence is a cascade of further, secondary extinctions. While a significant amount is known about the types of communities and species that suffer secondary extinctions, little is known about the consequences of secondary extinctions for biodiversity. Here we examine the effect of these secondary extinctions on trophic diversity, the range of trophic roles played by the species in a community. Our analyses of natural and model food webs show that secondary extinctions cause loss of trophic diversity greater than that expected from chance, a result that is robust to variation in food web structure, distribution of interactions strengths, functional response, and adaptive foraging. Greater than expected loss of trophic diversity occurs because more trophically unique species are more vulnerable to secondary extinction. This is not a straightforward consequence of these species having few links with others but is a complex function of how direct and indirect interactions affect species persistence. A positive correlation between a species' extinction probability and the importance of its loss defines high-risk species and should make their conservation a priority. PMID- 18419568 TI - A model for the evolution of assortative mating. AB - Abstract: Many animals and plants show a correlation between the traits of the individuals in the mating pair, implying assortative mating. Given the ubiquity of assortative mating in nature, why and how it has evolved remain open questions. Here we attempt to answer these questions in those cases where the trait under assortment is the same in males and females. We consider the most favorable scenario for assortment to evolve, where the same trait is under assortment and viability selection. We find conditions for assortment to evolve using a multilocus formalism in a haploid population. Our results show how epistasis in fitness between the loci that control the focal trait is crucial for assortment to evolve. We then assume specific forms of assortment in haploids and diploids and study the limiting cases of selective and nonselective mating. We find that selection for increased assortment is weak and that where increased assortment is costly, it does not invade. PMID- 18419569 TI - Life-history differences among coral reef sponges promote mutualism or exploitation of mutualism by influencing partner fidelity feedback. AB - Mutualism can be favored over exploitation of mutualism when interests of potential heterospecific partners are aligned so that individual organisms are beneficial to each others' continued growth, survival, and reproduction, that is, when exploitation of a particular partner individual is costly. A coral reef sponge system is particularly amenable to field experiments probing how costs of exploitation can be influenced by life-history characteristics. Pairwise associations among three of the sponge species are mutually beneficial. A fourth species, Desmapsamma anchorata, exploits these mutualisms. Desmapsamma also differs from the other species by growing faster, fragmenting more readily, and suffering higher mortality rates. Evaluating costs and benefits of association in the context of the complex life histories of these asexually fragmenting sponges shows costs of exploitation to be high for the mutualistic species but very low for this essentially weedy species. Although it benefits from association more than the mutualist species, by relying on their superior tensile strength and extensibility to reduce damage by physical disturbance, exploitation is favored because each individual host is of only ephemeral use. These sponges illustrate how life-history differences can influence the duration of association between individuals and, thus, the role of partner fidelity in promoting mutualism. PMID- 18419570 TI - Sexually selected traits predict patterns of species richness in a diverse clade of suboscine birds. AB - Whether sexual selection acts as an "engine of speciation" is controversial. Some studies suggest that it promotes the evolution of reproductive isolation, while others find no relationship between sexual selection and species richness. However, the explanatory power of previous models may have been constrained because they employed coarse-scale, between-family comparisons and used mating systems and morphological cues as surrogates for sexual selection. In birds, an obvious missing predictor is song, a sexually selected trait that functions in mate choice and reproductive isolation. We investigated the extent to which plumage dichromatism and song structure predicted species richness in a diverse family of Neotropical suboscine birds, the antbirds (Thamnophilidae). These analyses revealed a positive relationship between the intensity of sexual selection and diversity: genera with higher levels of dichromatism and lower pitched, more complex songs contained greater numbers of species. This relationship held when controlling for phylogeny and was strengthened by the inclusion of subspecies, suggesting that sexual selection has played a role in the diversification of antbirds. This is the first study to reveal correlations between song structure and species diversity, emphasizing the importance of acoustic signals, and within-family analyses, in comparative studies of sexual selection. PMID- 18419571 TI - Revisiting a model of ontogenetic growth: estimating model parameters from theory and data. AB - The ontogenetic growth model (OGM) of West et al. provides a general description of how metabolic energy is allocated between production of new biomass and maintenance of existing biomass during ontogeny. Here, we reexamine the OGM, make some minor modifications and corrections, and further evaluate its ability to account for empirical variation on rates of metabolism and biomass in vertebrates both during ontogeny and across species of varying adult body size. We show that the updated version of the model is internally consistent and is consistent with other predictions of metabolic scaling theory and empirical data. The OGM predicts not only the near universal sigmoidal form of growth curves but also the M(1/4) scaling of the characteristic times of ontogenetic stages in addition to the curvilinear decline in growth efficiency described by Brody. Additionally, the OGM relates the M(3/4) scaling across adults of different species to the scaling of metabolic rate across ontogeny within species. In providing a simple, quantitative description of how energy is allocated to growth, the OGM calls attention to unexplained variation, unanswered questions, and opportunities for future research. PMID- 18419572 TI - Sympatric speciation in birds is rare: insights from range data and simulations. AB - Sympatric speciation is now accepted as theoretically plausible and a likely explanation for divergence in a handful of taxa, but its contribution to large scale patterns of speciation remains contentious. A major problem is that it is difficult to differentiate between alternate scenarios of geographic speciation when species ranges have shifted substantially in the past. Previous studies have searched for a signal of the geographic mode of speciation by testing for a correlation between time since speciation and range overlap. Here we use simulations to show that the proportion of species showing zero or complete range overlap are more reliable indicators of the geography of speciation than is the correlation between time since speciation and overlap. We then apply these findings to the distributions of 291 pairs of avian sister species. Although 49% of pairs show some overlap in their ranges, our simulations show that this is not surprising under allopatric models of speciation. More revealingly, less than 2% show complete range overlap. Our simulations demonstrate that the observed patterns are most consistent with a model in which allopatric speciation is dominant but in which sympatric speciation is also present and contributes 5% of speciation events. PMID- 18419573 TI - Complex positive connections between functional groups are revealed by neural network analysis of ecological time series. AB - The relationships between functional linkages within communities and community dynamics are fundamental to biodiversity-stability relationships. By teasing apart the hidden layers within artificial neural networks (ANNs), we developed webs defining how functional groups influence each other's temporal dynamics. ANNs were based on 15 years of bimonthly monitoring of macrobenthic communities on three intertidal sandflats in Manukau Harbor (New Zealand). Sites differed in web topology and diversity, with the site dominated by one functional group exhibiting only a few strong links, the lowest alpha-, beta-, and gamma diversity, and the highest temporal stability in alpha-diversity. However, positive interactions between functional groups, nonconcordant with harborwide or site-specific environmental variables, always dominated the interaction webs. The increased number of links we observed with increased temporal variation of species richness within functional groups and overall diversity supports the insurance hypothesis. While our findings suggest that there may be no consistent model characterizing the topology of temporal interactions between functional groups, decreasing diversity is likely to decouple interactions between functional groups and decrease ecosystem functionality. PMID- 18419574 TI - Selection on herbivory resistance and growth rate in an invasive plant. AB - The evolution of increased competitive ability (EICA) hypothesis proposes that invasive species evolve decreased defense and increased competitive ability following natural enemy release. Previous tests of EICA examined the result of evolution by comparing individuals from home and introduced ranges, but no previous study of this hypothesis has examined the process of evolution by analyzing patterns of selection. On the basis of EICA, there should be selection for competitive ability without herbivores and selection for defense with herbivores. Selection on competitive ability should be stronger for genotypes accustomed to herbivores (home range genotypes), and selection on defense should be stronger for genotypes unaccustomed to herbivores (introduced range genotypes). Using a field experiment, we tested these hypotheses for the invasive plant Melaleuca quinquenervia. There was a negative genetic correlation between resistance and growth, indicating a trade-off. However, selection for stem elongation (an indicator of competitive ability) was always positive, and selection on resistance was always negative and did not depend on genotype source or the presence of herbivores. The patterns of selection found in this study contrast with predictions from EICA and accurately predict the lack of evolutionary change in growth and resistance following the introduction of this species from Australia to Florida. PMID- 18419575 TI - Complex copulatory behavior and the proximate effect of genital and body size differences on mechanical reproductive isolation in the millipede genus Parafontaria. AB - The role of species-specific genitalia in reproductive isolation is unclear. Males of the millipede genus Parafontaria use gonopods (modified eighth legs) charged with sperm from the genital openings of the second legs as intromittent organs. Males perform both preliminary and true intromission during mating. During preliminary intromission, a male attempts to insert his gonopods into the female genitalia before charging the gonopods with sperm. If this intromission is completed, it is followed by the ejaculation of sperm to the gonopods and true intromission for insemination. In two sympatric species of Parafontaria that lack effective precopulatory isolation, copulation was terminated without insemination because of preliminary intromission failure caused by mismatched genital and body sizes. Thus, mechanical isolation between these sympatric species resulted from morphological differentiation mediated by the obligatory preliminary intromission. These findings demonstrate the proximate importance of genital and body size differences for reproductive isolation within this genus of millipede. PMID- 18419576 TI - Widespread sequence variation in Epstein-Barr virus nuclear antigen 1 influences the antiviral T cell response. AB - Epstein-Barr virus (EBV) nuclear antigen (EBNA) 1 is perhaps the most widely studied EBV protein, because of its critical role in maintaining the EBV episome and its expression in all EBV-associated malignancies. Much of this research has focused exclusively on the EBV wild-type (wt) strain (B95-8). Sequence analysis of the gene encoding for EBNA1 in EBV isolates from 43 Caucasians has now revealed considerable EBNA1 sequence divergence from the EBV wt strain in the majority of isolates from this population group. Importantly, T cell recognition of an endogenously processed HLA-B8 - binding EBNA1 epitope was greatly influenced by this sequence polymorphism. PMID- 18419577 TI - Reemergence of recombinant vaccine-derived poliovirus outbreak in Madagascar. AB - BACKGROUND: After the 2001-2002 poliomyelitis outbreak due to recombinant vaccine derived polioviruses (VDPVs) in the Toliara province of Madagascar, another outbreak reoccurred in the same province in 2005. METHODS: We conducted epidemiological and virological investigations for each polio case patient and for their contacts. RESULTS: From May to August 2005, a total of 5 cases of acute flaccid paralysis were reported among unvaccinated or partially vaccinated children 2-3 years old. Type-3 or type-2 VDPV was isolated from case patients and from healthy contacts. These strains were classified into 4 recombinant lineages that showed complex mosaic genomic structures originating from different vaccine strain serotypes and probably from human enterovirus C (HEV-C) species. Genetic relatedness could be observed among these 4 lineages. Vaccination coverage of the population was very low (<50%). CONCLUSIONS: The broad distribution of VDPVs in the province and their close genetic relationship indicate intense and rapid cocirculation and coevolution of the vaccine strains and of their related HEV-C strains. The occurrence of an outbreak due to VDPV 3 years after a previous outbreak indicates that a short period with low vaccination coverage is enough to create favorable conditions for the emergence of VDPV in this setting. PMID- 18419578 TI - Reduced mother-to-child transmission of HIV associated with infant but not maternal GB virus C infection. AB - BACKGROUND: Prolonged coinfection with GB virus C (GBV-C) has been associated with improved survival in human immunodeficiency virus (HIV)-infected adults. We investigated whether maternal or infant GBV-C infection was associated with mother-to-child transmission (MTCT) of HIV-1 infection. METHODS: The study population included 1364 HIV-infected pregnant women enrolled in 3 studies of MTCT of HIV in Bangkok, Thailand (the studies were conducted from 1992-1994, 1996 1997, and 1999-2004, respectively). We tested plasma collected from pregnant women at delivery for GBV-C RNA, GBV-C antibody, and GBV-C viral genotype. If GBV C RNA was detected in the maternal samples, the 4- or 6-month infant sample was tested for GBV-C RNA. The rates of MTCT of HIV among GBV-C-infected women and infants were compared with the rates among women and infants without GBV-C infection. RESULTS: The prevalence of GBV-C RNA in maternal samples was 19%. Of 245 women who were GBV-C RNA positive, 101 (41%) transmitted GBV-C to their infants. Of 101 infants who were GBV-C RNA positive, 2 (2%) were infected with HIV, compared with 162 (13%) of 1232 infants who were GBV-C RNA negative (odds ratio [OR] adjusted for study, 0.13 [95% confidence interval {CI}, 0.03-0.54]). This association remained after adjustment for maternal HIV viral load, receipt of antiretroviral prophylaxis, CD4(+) count, and other covariates. MTCT of HIV was not associated with the presence of GBV-C RNA (adjusted OR [aOR], 0.94 [95% CI, 0.62-1.42]) or GBV-C antibody (aOR, 0.90 [95% CI, 0.54-1.50]) in maternal samples. CONCLUSIONS: Reduced MTCT of HIV was significantly associated with infant acquisition of GBV-C but not with maternal GBV-C infection. The mechanism for this association remains unknown. PMID- 18419579 TI - HIV and GB virus C infections seen from the perspective of the vertically coexposed infant. PMID- 18419580 TI - Association of human DNA helicase RecQ5beta with RNA polymerase II and its possible role in transcription. AB - Although RecQ5beta is a ssDNA (single-stranded DNA)-stimulated ATPase and an ATP dependent DNA helicase with strand-annealing activities, its cellular function remains to be explored. In the present paper, we used immunopurification and MS based analyses to show that human DNA helicase RecQ5beta is associated with at least four RNAP II (RNA polymerase II) subunits. RecQ5beta was also present in complexes immunoprecipitated using three different antibodies against the large subunit of RNAP II, or in complexes immunoprecipitated using an anti-FLAG antibody against either FLAG-RNAP II 33 kDa subunit or FLAG-Pin1. Different regions of the non-helicase domain of the RecQ5beta molecule were associated with hypophosphorylated and hyperphosphorylated forms of the RNAP II large subunit independently of DNA and RNA. RecQ5beta was also found in nuclear chromatin fractions and associated with the coding regions of the LDL (low-density lipoprotein) receptor and beta-actin genes. Knockdown of the RecQ5beta transcript increased the transcription of those genes. The results of the present study suggest that RecQ5beta has suppressive roles in events associated with RNAP II dependent transcription. PMID- 18419581 TI - MAP1 structural organization in Drosophila: in vivo analysis of FUTSCH reveals heavy- and light-chain subunits generated by proteolytic processing at a conserved cleavage site. AB - The MAP1 (microtubule-associated protein 1) family is a class of microtubule binding proteins represented by mammalian MAP1A, MAP1B and the recently identified MAP1S. MAP1A and MAP1B are expressed in the nervous system and thought to mediate interactions of the microtubule-based cytoskeleton in neural development and function. The characteristic structural organization of mammalian MAP1s, which are composed of heavy- and light-chain subunits, requires proteolytic cleavage of a precursor polypeptide encoded by the corresponding map1 gene. MAP1 function in Drosophila appears to be fulfilled by a single gene, futsch. Although the futsch gene product is known to share several important functional properties with mammalian MAP1s, whether it adopts the same basic structural organization has not been addressed. Here, we report the identification of a Drosophila MAP1 light chain, LC(f), produced by proteolytic cleavage of a futsch-encoded precursor polypeptide, and confirm co-localization and co-assembly of the heavy chain and LC(f) cleavage products. Furthermore, the in vivo properties of MAP1 proteins were further defined through precise MS identification of a conserved proteolytic cleavage site within the futsch-encoded MAP1 precursor and demonstration of light-chain diversity represented by multiple LC(f) variants. Taken together, these findings establish conservation of proteolytic processing and structural organization among mammalian and Drosophila MAP1 proteins and are expected to enhance genetic analysis of conserved MAP1 functions within the neuronal cytoskeleton. PMID- 18419582 TI - High Cu(I) and low proton affinities of the CXXC motif of Bacillus subtilis CopZ. AB - CopZ, an Atx1-like copper chaperone from the bacterium Bacillus subtilis, functions as part of a complex cellular machinery for Cu(I) trafficking and detoxification, in which it interacts specifically with the transmembrane Cu(I) transporter CopA. Here we demonstrate that the cysteine residues of the MXCXXC Cu(I)-binding motif of CopZ have low proton affinities, with both exhibiting pK(a) values of 6 or below. Chelator competition experiments demonstrated that the protein binds Cu(I) with extremely high affinity, with a small but significant pH-dependence over the range pH 6.5-8.0. From these data, a pH corrected formation constant, beta(2)= approximately 6 x 10(22) M(-2), was determined. Rapid exchange of Cu(I) between CopZ and the Cu(I)-chelator BCS (bathocuproine disulfonate) indicated that the mechanism of exchange does not involve simple dissociation of Cu(I) from CopZ (or BCS), but instead proceeds via the formation of a transient Cu(I)-mediated protein-chelator complex. Such a mechanism has similarities to the Cu(I)-exchange pathway that occurs between components of copper-trafficking pathways. PMID- 18419583 TI - Experimental hut evaluation of the pyrrole insecticide chlorfenapyr on bed nets for the control of Anopheles arabiensis and Culex quinquefasciatus. AB - OBJECTIVE: To determine the efficacy of chlorfenapyr against Anopheles arabiensis and Culex quinquefasciatus in East Africa and to identify effective dosages for net treatment in comparison with the commonly used pyrethroid deltamethrin. METHODS: Chlorfenapyr was evaluated on bed nets in experimental huts against A. arabiensis and C. quinquefasciatus in Northern Tanzania, at application rates of 100-500 mg/m(2). RESULTS: In experimental huts, mortality rates in A. arabiensis were high (46.0-63.9%) for all dosages of chlorfenapyr and were similar to that of deltamethrin-treated nets. Mortality rates in C. quinquefasciatus were higher for chlorfenapyr than for deltamethrin. Despite a reputation for being slow acting, >90% of insecticide-induced mortality in laboratory tunnel tests and experimental huts occurred within 24 h, and the speed of killing was no slower than for deltamethrin-treated nets. CONCLUSIONS: Chlorfenapyr induced low irritability and knockdown, which explains the relatively small reduction in blood-feeding rate. Combining chlorfenapyr with a more excito-repellent pyrethroid on bed nets for improved personal protection, control of pyrethroid resistant mosquitoes and pyrethroid resistance management would be advantageous. PMID- 18419584 TI - Interpretation of mycobacterial antibodies in the cerebrospinal fluid of adults with tuberculous meningitis. AB - OBJECTIVE: Microbiological identification of Mycobacterium tuberculosis is insensitive and slow, and clinical distinction of tuberculous meningitis (TBM) from other subacute or chronic meningoenchephalitides (SACM) is difficult. Successful use of highly specific M. tuberculosis serological assays on cerebrospinal fluid has been reported, but their performance for diagnosis in a tuberculosis endemic country where they would be of most value is unclear. We sought to determine the biological basis for the uncertainty in interpretation of antibody detection in the CSF of TBM patients. METHODS: We identified prospectively 46 adults with SACM and explored the concordance between TBM diagnosis and detection of highly specific M. tuberculosis antibodies in CSF. The source of antibodies in CSF was explored by evaluating the correlation between antibody titres in CSF with those in serum, or with the albumin quotient. Intrathecal IgG synthesis was assessed by the IgG index. RESULTS: Positive antibody titres were more frequent among TBM patients (76%), but were also present in individuals with other SACM (59%). A positive correlation between antibody titres in CSF with those in serum, or with the albumin quotient, supported the leakage of antibodies from plasma to CSF through an increased blood brain barrier permeability. Intrathecal IgG synthesis was only detected in 35% of the TBM cases. CONCLUSION: Plasma antibodies likely synthesized in response to previous tuberculosis infections were a major source of mycobacterial antibodies in CSF due to leakage through an impaired blood-brain barrier. Interpretation of mycobacterial antibodies in CSF of adults for TBM, however specific, must take into account the contribution of antibodies from plasma, and hence, has questionable use for diagnosis. PMID- 18419585 TI - Operational lessons from 20 years of the Mectizan Donation Program for the control of onchocerciasis. AB - The donation of ivermectin (Mectizan, Merck & Co., Inc.) to control onchocerciasis (river blindness) was established in 1987 and has since gradually expanded to provide for >570 million treatments cumulatively over the past 20 years. The Mectizan Donation Program (MDP) operates within a broad partnership in 33 endemic countries in need of mass treatment. Particular operational methods and tools are applied to facilitate ivermectin mass treatment. Drug management has been streamlined, including dosing, tablet size and packaging, and monitoring for adverse events. Much of the experience gained in the development of ivermectin mass treatment can be usefully applied in the recent broader perspective of control of neglected tropical diseases. The most important operational lessons of the MDP include: (i) the need to easily define the target population for treatment using rapid, non-invasive techniques; (ii) the value of a broad partnership; (iii) the great potential of working through community directed treatment; (iv) the need to streamline all drug management aspects and (v) the importance of operations research to tackle new challenges. PMID- 18419586 TI - Age and gender effects in self-reported urinary schistosomiasis in Tanzania. AB - OBJECTIVES: To characterize age-gender prevalence profiles of urinary schistosomiasis according to the questionnaire responses, compare the profiles to field survey data from selected regions, and determine if the profiles varied spatially throughout Tanzania. METHODS: In 2004, a national school-based questionnaire survey for self-reported schistosomiasis and blood in urine (BIU) was conducted in all regions of mainland Tanzania, to assist targeted mass distribution of praziquantel. Field survey data were collected in six north western and five coastal regions using microscopic examination of urine samples for the presence of Schistosoma haematobium eggs and assessment of micro haematuria with chemical reagent strips. Bayesian logistic regression models were created to calculate age-gender profiles adjusted for demographic and ecological covariates and spatial correlation in the questionnaire data. Separate odds ratios (OR) for age-gender effects were calculated in each administrative area. RESULTS: Data were obtained from > 2.5 million schoolchildren. Boys had higher prevalence of self-reported schistosomiasis and BIU than girls. In boys, prevalence according to the questionnaire and field surveys followed similar age profiles. However, in girls, prevalence according to the field surveys increased in older age groups, but flattened out or decreased according to the questionnaire, indicating the latter underestimated prevalence in older girls. In the models, little spatial correlation was evident in the OR for the age-gender effects, suggesting that these did not vary spatially. CONCLUSION: Age-gender patterns of urinary schistosomiasis were consistent in different geographical areas of Tanzania. Because the questionnaire underestimated prevalence in older girls, we propose that upward calibration of observed prevalence is done for older females only. PMID- 18419587 TI - Exemplars in the mist: the cognitive substrate of the representativeness heuristic. AB - The idea that people often make probability judgments by a heuristic short-cut, the representativeness heuristic, has been widely influential, but also criticized for being vague. The empirical trademark of the heuristic is characteristic deviations between normative probabilities and judgments (e.g., the conjunction fallacy, base-rate neglect). In this article the authors contrast two hypotheses concerning the cognitive substrate of the representativeness heuristic, the prototype hypothesis (Kahneman & Frederick, 2002) and the exemplar hypothesis (Juslin & Persson, 2002), in a task especially designed to elicit representativeness effects. Computational modelling and an experiment reveal that representativeness effects are evident early in training and persist longer in a more complex task environment and that the data are best accounted for by a model implementing the exemplar hypothesis. PMID- 18419588 TI - Comparison of chemosensory, auditory and visual event-related potential amplitudes. AB - To enhance understanding of neuropsychological processing underlying chemosensory event-related potentials (CSERPs), the objective of the present study was to compare CSERPs with auditory (AERPs) and visual event-related potentials (VERPs), and to approach the question of similarities in neuropsychological processing by means of correlation and cluster analyses. A multisensory ERP protocol was used, in which olfactory/chemosomatosensory (pyridine), auditory (1000-Hz tone) and visual (white circle) stimuli were presented in a pseudo-randomized sequence to 18 young, healthy adults. The results show (1) a morphological relation between P2 and P3 components that is different for the chemosensory than for the auditory and visual modalities, (2) a relatively weak association between the chemosensory and auditory/visual modalities for P3 in terms of both correlation and cluster, and (3) a P2/P3 cluster for the chemosensory but not auditory or visual modality. This supports the claim that the chemosensory P2 and P3 components reflect similar neuropsychological processes. PMID- 18419589 TI - A comparison of normative data for the Trail Making Test from several countries: equivalence of norms and considerations for interpretation. AB - The Trail Making Test may not be equivalent across cultures, i.e., differences in the scores across different cultures may not reveal real differences in the ability of the subjects on the construct being measured. In order to assess this hypothesis, normative samples from ten different countries were compared. Age decade subgroups across samples were ranked based on mean time taken to complete each part of the task. Large Z scores differences were found between these samples when comparing the first with the second, and the last in the rank. These differences were significant even when age and education were comparable across samples. Following Van de Vijver & Tanzer (1997), several possible sources of bias were identified. Incomparability of samples and administration differences were the most likely factors accounting for differences. Because of the lack of validity studies in the countries considered, no firm conclusions could be obtained regarding construct bias. Although the TMT may be measuring visual scanning, psychomotor speed and mental flexibility, normative data from different countries and cultures are not equivalent which might lead to serious diagnostic errors. PMID- 18419590 TI - Mothers' representations of caregiving and their adult children's representations of attachment: intergenerational concordance and relations to beliefs about mothering. AB - Mothers (N= 35) and their adult children completed questionnaires and were interviewed in order to examine relationships between mothers' caregiving representations and their adult children's attachment representations, and relationships between attachment/caregiving representations and beliefs about mothering. Mothers' and their children's accounts of and present thinking about their past relationship were highly similar, indicating that the two parts develop concordant states of mind regarding their relationship. In contrast, there was no relationship between mothers' and their adult children's beliefs about mothering, suggesting that such beliefs are not simply passed on from generation to generation within families. Attachment/caregiving classification interacted with generation in influencing a belief that biological facts determine maternal behavior, young adults with preoccupied attachment being particularly prone to reject this idea. Attachment/caregiving classification also had a significant effect on participants' tendency to adhere to an idealized conception of mothering, this tendency being associated with a dismissive attachment/caregiving representation. PMID- 18419591 TI - Free the animals? Investigating attitudes toward animal testing in Britain and the United States. AB - In this study, 185 British and 143 American undergraduates completed a battery of tests that measured attitudes toward animal testing and various individual difference variables. Attitudes toward animal testing factored into two interpretable factors: general attitudes toward animal testing, and animal welfare and conditions of testing. Overall, there was support for animal testing under the right conditions, although there was also concern for the welfare of animals and the conditions under which testing takes place. There were small but significant national difference on both factors (with Americans more positive about testing and less positive about animal welfare), and a significant sex difference on the first factor (women were more negative about testing). Correlation and regression analyses showed that there were few significant individual difference predictors of both factors. These results are discussed in relation to past and future work on attitudes toward animal testing. PMID- 18419592 TI - Personality types and risky health behaviors in Norwegian students. AB - Typological research has received increasing interest in recent years, but is still regarded as substandard by personality psychologists. The current investigation shows how a typological approach can enhance our understanding of the influence of personality on risky health behaviors. We focused on Torgersen's eight personality types that represent unique configurations of high and low Extraversion, Neuroticism, and Constraint (Vollrath & Torgersen, 2000). Participants were 606 Norwegian university students. Measures assessed personality factors, smoking, abuse of drugs and alcohol, drunk driving, and risky sexual behaviors. Results replicated earlier research showing that types combining either high Extraversion and low Constraint (Hedonists, Impulsives) or high Neuroticism and low Constraint (Insecures) engage in more risky health behaviors. The discussion focuses on demonstrating that the study of types is a valuable supplement to the dimensional tradition in personality psychology. PMID- 18419593 TI - Effects of the premenstrual syndrome on facial expressions of sadness. AB - This study examined the relationship between PMS and emotion-related electromyographic facial activity at different phases of the menstrual cycle. Twenty-four women of reproductive age (12 with PMS, 12 controls) participated in two EMG sessions (T1 and T2) in which they were shown photographic images that can elicit various emotions (IAPS stimuli). T1 took place in the follicular phase, T2 in the luteal phase. The activity of the musculus depressor anguli oris ("depressor", expression of sadness) was measured. Depressor activity was compared to activity of musculus orbicularis oculi ("orbicularis"; expression of joy). ANOVA yielded a significant increase of the activity of the depressor at T2 in the PMS group. The PMS group showed more frequent depressor activity during the luteal than the follicular phase. Orbicularis activity did not change from T1 to T2. CONCLUSIONS: The PMS group experienced various visual stimuli in a more depressed way during the luteal phase. PMID- 18419594 TI - Early homing of adult mesenchymal stem cells in normal and infarcted isolated beating hearts. AB - Little is known on the early homing features of transplanted mesenchymal stem cells (MSCs). We used the isolated rat heart model to study the homing of MSCs injected in the ventricular wall of a beating heart. In this model all types of cells and matrix elements with their interactions are represented, while external interferences by endothelial/neutrophil interaction and neurohormonal factors are excluded. We studied the morphology and marker expression of MSCs implanted in normal hearts and in the border-zone of infarcted myocardium. Early morphological adaptation of MSC homing differs between normal and infarcted hearts over the first 6 hrs after transplantation. In normal hearts, MSCs migrate very early through the interstitial milieu and begin to show morphological changes. Yet, in infarcted hearts MSCs remain in the site of injection forming clusters of round shaped cells in the border-zone of the infarcted area. Both in normal and infarcted hearts, immuno-histochemistry and confocal imaging showed that, besides the proliferative marker proliferating cell nuclear agent (PCNA), some transplanted cells early express myoblastic maker GATA-4, and some of them show a VWF immunopositivity. Moreover, a few hours after injection connexin-43 is well evident between cardiomyocytes and injected cells. This study indicates for the first time that the isolated beating heart is a good model to study early features of MSC homing without external interferences. The results show (i) that MSCs start to change marker expression few hours after injection into a beating heart and (ii) that infarcted myocardium influences transplanted MSC morphology and mobility within the heart. PMID- 18419595 TI - Opposite effects of uracil and adenine nucleotides on the survival of murine cardiomyocytes. AB - We previously showed that the human heart expresses all known P2X and P2Y receptors activated by extra-cellular adenine or uracil nucleotides. Despite evidence that, both in humans and rodents, plasma levels of ATP and UTP markedly increase during myocardial infarction, the differential effects mediated by the various adenine- and uracil-preferring myocardial P2 receptors are still largely unknown. Here, we studied the effects of adenine and uracil nucleotides on murine HL-1 cardiomyocytes. RT-PCR analysis showed that HL-1 cardiomyocytes express all known P2X receptors (except for P2X(2)), as well as the P2Y(2,4,6,14) subtypes. Exposure of cardiomyocytes to adenine nucleotides (ATP, ADP or BzATP) induced apoptosis and necrosis, as determined by flow-cytometry. Cell death was exacerbated by tumour necrosis factor (TNF)-alpha, a cytokine implicated in chronic heart failure progression. Conversely, uracil nucleotides (UTP, UDP and UDPglucose) had no effect 'per se', but fully counteracted the deleterious effects induced by adenine nucleotides and TNF-alpha, even if added to cardiomyocytes after beginning exposure to these cell death-inducing agents. Thus, exposure of cardiomyocytes to elevated concentrations of ATP or ADP in the presence of TNF-alpha contributes to cell death, an effect which is counteracted by uracil-preferring P2 receptors. Cardiomyocytes do not need to be 'primed' by uracil nucleotides to become insensitive to adenine nucleotides-induced death, suggesting the existence of a possible 'therapeutic' window for uracil nucleotides-mediated protection. Thus, release of UTP during cardiac ischaemia and in chronic heart failure may protect against myocardial damage, setting the basis for developing novel cardioprotective agents that specifically target uracil-preferring P2Y receptors. PMID- 18419596 TI - Researching into the cellular shape, volume and elasticity of mesenchymal stem cells, osteoblasts and osteosarcoma cells by atomic force microscopy. AB - Within the bone lie several different cell types, including osteoblasts (OBs) and mesenchymal stem cells (MSCs). The MSCs are ideal targets for regenerative medicine of bone due to their differentiation potential towards OBs. Human MSCs exhibit two distinct morphologies: rapidly self-renewing cells (RS) and flat cells (FC) with very low proliferation rates. Another cell type found in pathological bone conditions is osteosarcoma. In this study, we compared the topographic and morphometric features of RS and FC cells, human OBs and MG63 osteosarcoma cells by atomic force microscopy (AFM). The results demonstrated clear differences: FC and hOB cells showed similar ruffled topography, whereas RS and MG63 cells exhibited smoother surfaces. Furthermore, we investigated how selected substrates influence cell morphometry. We found that RS and MG63 cells were flatter on fibrous substrates such as polystyrene and collagen I, but much more rounded on glass, the smoothest surface. In contrast, cells with large area, namely FC and hOB cells, did not exhibit pronounced changes in flatness with regards to the different substrates. They were, however, remarkably flatter in comparison to RS and MG63 cells. We could explain the differences in flatness by the extent of adhesion. Indeed, FC and hOB cells showed much higher content of focal adhesions. Finally, we used the AFM to determine the cellular Young's modulus. RS, FC and hOB cells showed comparable stiffness on the three different substrates, while MG63 cells demonstrated the unique feature of increased elasticity on collagen I. In summary, our results show, for the first time, a direct comparison between the morphometric and biophysical features of different human cell types derived from normal and pathological bone. Our study manifests the opinion that along with RNA, proteomic and functional research, morphological and biomechanical characterization of cells also reveals novel cell features and interrelationships. PMID- 18419597 TI - High-resolution analysis of genetic stability of human adipose tissue stem cells cultured to senescence. AB - The potential use of human mesenchymal stem cells for therapeutic applications implies large scale in vitro culture, increasing the probability of genetic instability and transformation. We examine here the incidence of unbalanced and balanced chromosome rearrangements in polyclonal and single cell-derived cultures of human adipose stem cells to senescence. G-banding karyotyping of the polyclonal cultures shows a normal karyotype. In addition, high-resolution microarray-based comparative genomic hybridization analyses relative to uncultured adipose stem cells from the same donors reveal overall genomic stability in long-term (approximately 6 months) polyclonal and clonal culture. One adipose stem cell clone displayed minor deletions in gene-rich telomeric and sub-telomeric regions on three chromosomes in early passage. This however, was detected only in a sub-population of cells that was subsequently spontaneously eliminated from the culture. Apparent pericentromeric instabilities are also occasionally detected in specific chromosomes. Our results indicate that clonal chromosomal aberrations may arise transiently in early passage adipose stem cells (ASC) cultures. Nonetheless, incidence of these aberrations seems to be negligible in the majority of long-term ASC cultures, at least under the culture conditions used here. PMID- 18419598 TI - Correlation between the expression of divalent metal transporter 1 and the content of hypoxia-inducible factor-1 in hypoxic HepG2 cells. AB - Transferrin and transferrin receptor are two key proteins of iron metabolism that have been identified to be hypoxia-inducible genes. Divalent metal transporter 1 (DMT1) is also a key transporter of iron under physiological conditions. In addition, in the 5' regulatory region of human DMT1 (between -412 and -570), there are two motifs (CCAAAGTGCTGGG) that are similar to hypoxia-inducible factor 1 (HIF-1) binding sites. It was therefore speculated that DMT1 might also be a hypoxia-inducible gene. We investigated the effects of hypoxia and hypoxia/re oxygenation on the expression of DMT1 and the content of HIF-1alpha in HepG2 cells. As we expected, a very similar tendency in the responses of the expression of HIF-1alpha, DMT1+IRE (iron response element) and DMT1-IRE proteins to chemical (CoCl(2)) or physical hypoxia was observed. A highly significant correlation was found between the expression of DMT1 proteins and the contents of HIF-1 in hypoxic cells. After the cells were exposed to hypoxia and subsequent normoxia, no HIF-1alpha could be detected and a significant decrease in DMT1+IRE expression (P<0.05), but not in DMT1-IRE protein (versus the hypoxia group), was observed. The findings implied that the HIF-1 pathway might have a role in the regulation of DMT1+IRE expression during hypoxia. PMID- 18419599 TI - Pro-inflammatory effects of hydrogen sulphide on substance P in caerulein-induced acute pancreatitis. AB - Hydrogen sulphide (H(2)S), a novel gasotransmitter, has been recognized to play an important role in inflammation. Cystathionine-gamma-lyase (CSE) is a major H(2)S synthesizing enzyme in the cardiovascular system and DL-propargylglycine (PAG) is an irreversible inhibitor of CSE. Substance P (SP), a product of preprotachykinin-A (PPT-A) gene, is a well-known pro-inflammatory mediator which acts principally through the neurokinin-1 receptor (NK-1R). We have shown an association between H(2)S and SP in pulmonary inflammation as well as a pro inflammatory role of H(2)S and SP in acute pancreatitis. The present study was aimed to investigate the interplay between pro-inflammatory effects of H(2)S and SP in a murine model of caerulein-induced acute pancreatitis. Acute pancreatitis was induced in mice by 10 hourly intraperitoneal injections of caerulein (50 (g/kg). PAG (100 mg/kg, i.p.) was administered either 1 hr before (prophylactic) or 1 hr after (therapeutic) the first caerulein injection. PAG, given prophylactically as well as therapeutically, significantly reduced plasma H(2)S levels and pancreatic H(2)S synthesizing activities as well as SP concentrations in plasma, pancreas and lung compared with caerulein-induced acute pancreatitis. Furthermore, prophylactic as well as therapeutic administration of PAG significantly reduced PPT-A mRNA expression and NK-1R mRNA expression in both pancreas and lung when compared with caerulein-induced acute pancreatitis. These results suggest that the pro-inflammatory effects of H(2)S may be mediated by SP NK-1R pathway in acute pancreatitis. PMID- 18419600 TI - Trichostatin A causes p53 to switch oxidative-damaged colorectal cancer cells from cell cycle arrest into apoptosis. AB - Many studies aim at improving therapeutic efficacy by combining strategies with oxidative stress-inducing drugs and histone deacetylase (HDAC) inhibitors in colorectal cancer. As p53 and p21(WAF1) are essential in oxidative stress-induced DNA damage, we investigated epigenetic regulation of p21(WAF1) promoter. Firstly, HCT116 p53(+)/(+) and p53(-)/(-) colorectal cancer cells were treated with H(2)O(2) for 6 hrs and 24 hrs (early/late response). Chromatin immunoprecipitation revealed transcriptional transactivation of p21(WAF1) in HCT116 p53(+)/(+) cells as shown by increased binding of p53 and acetylated H4 around two p21(WAF1) promoter sites, the responsible element (RE) and the Sp1 site, while both proteins bound preferentially on the RE. Interestingly, H3 was not involved, suggesting H4-specific transactivation of the p21(WAF1) promoter. H(2)O(2) addition resulted in G(2)/M arrest of both HCT116 cell lines without significant cell death. To investigate whether a HDAC inhibitor strengthens G(2)/M arrest, we pretreated cells with Trichostatin A (TSA). In HCT116 p53(+)/(+) cells, we found (i) remarkably increased acetylated H4 around both p21(WAF1) promoter regions, especially at the Sp1 site; (ii) increased acetylation of p53 at lysines 320 and 382;(iii) displacement of HDAC1 from the Sp1 site, thus inhibiting its repression effect and increasing p53 binding.p53 seems to trigger H4-acetylation around the p21(WAF1) promoter because there was nearly no H4 acetylation in HCT116 p53(-)/(-) cells. For the first time we show that there is a time-dependent TSA mode of action with increased p53-dependent histone H4 acetylation at the p21(WAF1) promoter in early response, and decreased acetylation in late response. Reduced p53-triggered transactivation of p21(WAF1) in late response allows cells to re-enter cell cycle, and TSA causes p53 to simultaneously induce apoptosis. PMID- 18419601 TI - Consequences of altered eicosanoid patterns for nociceptive processing in mPGES-1 deficient mice. AB - Cyclooxygenase-2 (COX-2)-dependent prostaglandin (PG) E(2) synthesis in the spinal cord plays a major role in the development of inflammatory hyperalgesia and allodynia. Microsomal PGE(2) synthase-1 (mPGES-1) isomerizes COX-2-derived PGH(2) to PGE(2). Here, we evaluated the effect of mPGES-1-deficiency on the nociceptive behavior in various models of nociception that depend on PGE(2) synthesis. Surprisingly, in the COX-2-dependent zymosan-evoked hyperalgesia model, the nociceptive behavior was not reduced in mPGES-1-deficient mice despite a marked decrease of the spinal PGE(2) synthesis. Similarly, the nociceptive behavior was unaltered in mPGES-1-deficient mice in the formalin test. Importantly, spinal cords and primary spinal cord cells derived from mPGES-1 deficient mice showed a redirection of the PGE(2) synthesis to PGD(2), PGF(2alpha) and 6-keto-PGF(1alpha) (stable metabolite of PGI(2)). Since the latter prostaglandins serve also as mediators of nociception they may compensate the loss of PGE(2) synthesis in mPGES-1-deficient mice. PMID- 18419602 TI - Overexpression of vascular endothelial growth factor (VEGF) receptors on keratinocytes in psoriasis: regulated by calcium independent of VEGF. AB - Psoriasis is a common chronic inflammatory disease of the skin characterized by epidermal hyperplasia and angiogenesis. Recently, vascular endothelial growth factor receptors (VEGFRs, including VEGFR-1, VEGFR-2 and VEGFR-3) were found to be expressed in normal human epidermis and associated with proliferation and migration of keratinocytes. The purpose of this study is to investigate the expression of VEGFRs on psoriatic keratinocytes and the roles of calcium and VEGF in regulating VEGFR expression. Skin samples from 17 patients with chronic plaque psoriasis and 11 normal controls were included. The expression of VEGFRs in psoriatic keratinocytes at mRNA and protein levels was determined by reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blot analysis. Localization of the VEGFRs in skin lesions was determined by immuno-fluorescent method. Since keratinocyte proliferation and differentiation rely on calcium concentrations, and VEGF is overexpressed in psoriatic epidermis, we further investigated the roles of calcium and VEGF in regulating the expression of VEGFRs. Overexpression of VEGFR-1, VEGFR-2 and VEGFR-3 in psoriatic epidermis was demonstrated both at mRNA and protein levels in vitro. VEGFRs were strongly labeled in non-lesional, perilesional and lesional psoriatic keratinocytes in all viable epidermal stratums in vivo. Furthermore, both exogenous VEGF165 and calcium enhanced the expression of VEGFRs. Calcium also enhanced the expression of VEGF in non-lesional psoriatic keratinocytes, while targeted blockade of VEGF activity by bevacizumab could not inhibit calcium-induced up-regulation of protein levels of VEGFRs. We conclude from these results that VEGFRs are overexpressed in lesional psoriatic epidermal keratinocytes. Both calcium and VEGF regulate VEGFRs expression in psoriatic epidermis. More importantly, calcium is a potential regulator for VEGFR independent of VEGF. PMID- 18419603 TI - Selective CB2 up-regulation in women affected by endometrial inflammation. AB - Endometritis is defined as an inflammation of the endometrial mucosa of the uterus. In endometritis large amounts of toxic mediators, including nitric oxide (NO) are released by inflammatory cells. As a consequence of nitric oxide dependent injury, the cells respond by triggering protective mechanisms, by changing the endocannabinoid system (ECS) which comprises both CB(1) and CB(2) cannabinoid receptors and their endogenous ligands. The aim of our study was to seek out evidence for the presence of cannabinoid receptors in inflammatory endometrial tissue as well as for their potential role in endometrial inflammation. Our results showed a selective up-regulation of both transcription and expression of CB(2) receptors in biopsies from women affected by endometrial inflammation compared to healthy women. The experiments with the nitric oxide donor S-Nitroso-L-Glutathione (GSNO) suggest that such a selective up-regulation may be related to the nitric oxide release occurring during endometrial inflammation. In addition, we demonstrated an increase in chymase expression, a marker of mast cells, in biopsies of women affected by endometritis. Therefore our results support the hypothesis that the up-regulation of CB(2) occurs mainly on mast cells and that it might tend to sensitize these cells to the anti inflammatory effect exerted by endogenous cannabinoids by binding their receptor and thus preventing the mast cell degranulation and the release of pro inflammatory mediators. In conclusion, we believe that the selective CB(2) up regulation might play a role as a novel prognostic factor in endometrial inflammation. PMID- 18419604 TI - ARG3.1/ARC expression in hippocampal dentate gyrus astrocytes: ultrastructural evidence and co-localization with glial fibrillary acidic protein. AB - Synaptic efficacy following long-term potentiation (LTP) and memory consolidation is associated with changes in the expression of immediate early genes (IEGs). These changes are often accompanied by increased expression of glial fibrillary acidic protein (GFAP). While the protein products of the majority of IEGs are mainly restricted to the cell body, Arg3.1/Arc product is rapidly delivered to dendrites, where it accumulates close to synaptic sites. Arg3.1/Arc protein was originally considered neurone specific; however, we have recently found Arg3.1/Arc immunoreactivity (Arg3.1/Arc-IR) within glial cells and demonstrated its increased expression after LTP in the hippocampal dentate gyrus (DG). Here, we have further investigated this novel finding, using electron microscopic immunocytochemistry to determine the localization and sub-cellular distribution of Arg3.1/Arc protein in GFAP positive glia (GFAP-IR) in the DG. Arg3.1/Arc labelling was seen prominently in GFAP-IR glial cell bodies and in large- and medium-sized glial filamentous processes. GFAP-labelled medium-small peri synaptic glial profiles also displayed Arg3.1/Arc-IR; however, the very thin and distal glial filaments only displayed Arc-IR. Arc-IR was distributed throughout the cytoplasm, often associated with GFAP filaments, and along the plasma membrane of glial processes. Peri-synaptic glial Arg3.1/Arc-IR processes were apposed to pre- and/or post-synaptic profiles at asymmetric axospinous synapses. These data, taken with our earlier study which provided evidence for an increase in astrocytic Arg3.1/Arc-IR after the induction of LTP, suggest a role for glial Arg3.1/Arc in structural and synaptic plasticity which may be critical for the maintenance of cognitive functions. PMID- 18419606 TI - The anti-angiogenic 8-epipuupehedione behaves as a potential anti-leukaemic compound against HL-60 cells. AB - We have previously reported that 8-epipuupehedione, a synthetic derivative of sesquiterpenes found in several kinds of sponges, is a potent inhibitor of angiogenesis. Here, we show that 8-epipuupehedione is also a potent anti- leukaemic compound, targeting three hallmarks of malignancy: proliferation, survival and extra-cellular matrix re-modelling. To fulfil this goal, we use the HL-60 promyeolocytic cells as our model system and the following experimental procedures: cell growth assay, Hoetsch staining, cell cycle analysis and DNA fragmentation, caspase 3 activity and zymographic assays. Our results show that this compound inhibits proliferation and has potent and specific pro-apoptotic effects on HL-60 promyelocytic cells, inducing their nuclei and DNA fragmentation, as well as caspase 3 activity activation. Furthermore, 8 epipuupehedione strongly inhibits matrix metalloproteinase-2 and urokinase production by HL-60 cells. These results suggest that 8-epipuupehedione could be an attractive drug for further evaluation in the treatment of leukemia. PMID- 18419605 TI - Immunosuppression induced by immature dendritic cells is mediated by TGF-beta/IL 10 double-positive CD4+ regulatory T cells. AB - Dendritic cells (DC) have important functions in T cell immunity and T cell tolerance. Previously, it was believed that T cell unresponsiveness induced by immature DC (iDC) is caused by the absence of inflammatory signals in steady state in vivo conditions and by the low expression levels of costimulatory molecules on iDC. However, a growing body of evidence now indicates that iDC can also actively maintain peripheral T cell tolerance by the induction and/or stimulation of regulatory T cell populations. In this study, we investigated the in vitro T cell stimulatory capacity of iDC and mature DC (mDC) and found that both DC types induced a significant increase in the number of transforming growth factor (TGF)-beta and interleukin (IL)-10 double-positive CD4(+) T cells within 1 week of autologous DC/T cell co-cultures. In iDC/T cell cultures, where antigen specific T cell priming was significantly reduced as compared to mDC/T cell cultures, we demonstrated that the tolerogenic effect of iDC was mediated by soluble TGF-beta and IL-10 secreted by CD4(+)CD25(-)FOXP3(-) T cells. In addition, the suppressive capacity of CD4(+) T cells conditioned by iDC was transferable to already primed antigen-specific CD8(+) T cell cultures. In contrast, addition of CD4(+) T cells conditioned by mDC to primed antigen specific CD8(+) T cells resulted in enhanced CD8(+) T cell responses, notwithstanding the presence of TGF-beta(+)/IL-10(+) T cells in the transferred fraction. In summary, we hypothesize that DC have an active role in inducing immunosuppressive cytokine-secreting regulatory T cells. We show that iDC conditioned CD4(+) T cells are globally immunosuppressive, while mDC induce globally immunostimulatory CD4(+) T cells. Furthermore, TGF-beta(+)/IL-10(+) T cells are expanded by DC independent of their maturation status, but their suppressive function is dependent on immaturity of DC. PMID- 18419607 TI - Solar elastosis and presence of mast cells as key features in the pathogenesis of melasma. AB - Melasma is an acquired hypermelanosis on sun-exposed areas. Its pathogenesis has not been clearly elucidated. Using histochemistry (Giemsa, Verhoeff-van Gieson and Fontana-Masson staining), we evaluated melasma lesions and compared them with nonlesional skin. Skin samples were obtained from lesional and nonlesional facial skin of 27 patients with melasma, and biopsies were also taken from normal control subjects. Mast cells and solar elastosis areas were evaluated using a computer-assisted image-analysis program. Lesional skin had abundant elastotic material compared with nonaffected skin (13.3 +/- 2.8% vs. 10.2 +/- 2.9%, P < 0.001). Mast cells were more prominent in the elastotic areas of melasma skin (173 +/- 57% vs. 145 +/- 57%, P = 0.04). Melasma could be a result of a cumulative sun exposure, in a microenvironment of cutaneous photoageing in which inflammatory cells, particularly mast cells, play a key role. PMID- 18419608 TI - MicroRNAs: novel regulators in skin inflammation. AB - Compelling evidence indicates that microRNAs (miRNAs), short, non-protein coding RNAs, are critical for the development and survival of multicellular organisms. Recently, miRNAs were implicated in the pathogenesis of psoriasis and atopic eczema (AE), the two most common chronic inflammatory disorders in skin. In particular, miR-203, the first skin-specific miRNA, showing an intriguing expression profile being confined to skin epithelium, is specifically overexpressed in psoriasis. MiR-146a, another miRNA showing specific upregulation in psoriasis, is involved in the regulation of innate immune responses and the tumour necrosis factor (TNF)-alpha pathway. Interestingly, miR-125b, another miRNA involved in the TNF-alpha pathway, is also deregulated in psoriasis and AE. As skin inflammation may serve as a model for chronic inflammatory disorders, it is likely that miRNAs involved in skin inflammation will eventually emerge in other inflammatory or autoimmune disorders, and some of these may become disease markers and therapeutic targets. In this review we present an overview of what is currently known about the roles of miRNAs in chronic inflammatory skin disorders. PMID- 18419609 TI - Partial unilateral lentiginosis colocalized with naevus depigmentosus. PMID- 18419610 TI - Calciphylaxis in a diabetic patient provoked by warfarin therapy. PMID- 18419611 TI - Tjalma's or pseudo-pseudo-Meigs' syndrome: a case report. PMID- 18419612 TI - A clinicopathological surprise: amelanotic malignant melanoma. PMID- 18419613 TI - Oral ulcers in an immunosuppressed 5-year-old boy. PMID- 18419614 TI - Erythematous papules on the parasternal region in a 76-year-old man. PMID- 18419615 TI - Multiple papules affecting the axillae and groin. PMID- 18419616 TI - An unusual congenital lesion in a 26-year-old woman. PMID- 18419617 TI - A rare and challenging diagnosis not to be missed. PMID- 18419618 TI - Painful unusual ulcers of the ankle. PMID- 18419620 TI - Identification and management of undiagnosed and undertreated allergic rhinitis in adults and children. AB - Allergic rhinitis (AR) is a common health problem that affects adults, adolescents and children and is often undiagnosed or inadequately treated. Because AR is not a life-threatening disease, many patients do not seek medical treatment for their symptoms, and others self-medicate with over-the-counter medications, often sedating antihistamines. However, untreated or inadequately treated AR can substantially impair overall quality of life (QOL) by causing fatigue, headache, cognitive impairment and other problems. The risk for comorbid conditions, such as asthma, otitis media, and lymphoid hypertrophy with obstructive sleep apnea, can increase, and the symptoms of AR can worsen if AR is not adequately treated. Among the symptoms of AR, nasal congestion has been described by patients as the most bothersome because it disrupts sleep, resulting in diminished daytime performance. A new congestion screening tool, the Congestion Quantifier, has been developed to aid in the diagnosis and treatment of AR and to help guide treatment decisions. Intranasal corticosteroids (INSs) are recommended as effective pharmaceutical treatments for controlling the symptoms of AR. Randomized, controlled trials in children and adults have demonstrated that INSs relieve rhinitis symptoms, thereby improving QOL in individuals with seasonal or perennial AR. Most INSs are approved for use in children >or=6 years of age, but mometasone furoate and fluticasone furoate are approved for use in children as young as 2 years of age and fluticasone propionate for children >or=4 years old. Long-term benefits have also been seen with the use of immunotherapy, although some patients, especially children, resist the injections used in subcutaneous immunotherapy. Recent studies with sublingual immunotherapy have indicated that it might be an effective and well tolerated alternative to immunotherapy injections. PMID- 18419622 TI - Genes associated with the tumour microenvironment are differentially expressed in cured versus primary chemotherapy-refractory diffuse large B-cell lymphoma. AB - In order to identify genes associated with primary chemotherapy-resistance, gene expression profiles (GEP) in tumour tissue from 37 patients with de novo diffuse large B-cell lymphoma (DLBCL), stage II-IV, either in continuous complete remission (n = 24) or with progressive disease during primary treatment (n = 13), were examined using spotted 55K oligonucleotide arrays. Immunohistochemistry was used for confirmation at the protein level. The top 86 genes that best discriminated between the two cohorts were chosen for further analysis. Only seven of 86 genes were overexpressed in the refractory cohort, e.g. RABGGTB and POLE, both potential targets for drug intervention. Seventy-nine of 86 genes were overexpressed in the cured cohort and mainly coded for proteins expressed in the tumour microenvironment, many of them involved in proteolytic activity and remodelling of extra cellular matrix. Furthermore, major histocompatibility complex class I molecules, CD3D and ICAM1 were overexpressed, indicating an enhanced immunological reaction. Immunohistochemistry confirmed the GEP results. The frequency of tumour infiltrating lymphocytes, macrophages, and reactive cells expressing ICAM-1, lysozyme, cathepsin D, urokinase plasminogen activator receptor, signal transducer and activator of transcription 1, and galectin-3 was higher in the cured cohort. These findings indicate that a reactive microenvironment has an impact on the outcome of chemotherapy in DLBCL. PMID- 18419623 TI - Erythrocyte vesiculation: a self-protective mechanism? AB - Previous studies demonstrated that 20% of haemoglobin is lost from circulating erythrocytes during their total lifespan by vesiculation. To study whether removal molecules other than membrane-bound haemoglobin were present in erythrocyte-derived vesicles, flow cytometry and immunoblot analysis were employed to examine the presence of phosphatidylserine (PS) and IgG, and senescent cell antigens respectively. It was demonstrated that 67% of glycophorin A-positive vesicles exposed PS, and that half of these vesicles also contained IgG. Immunoblot analysis revealed the presence of a breakdown product of band 3 that reacted with antibodies directed against senescent erythrocyte antigen associated band 3 sequences. In contrast, only the oldest erythrocytes contained senescent cell antigens and IgG, and only 0.1% of erythrocytes, of all ages, exposed PS. It was concluded that vesiculation constitutes a mechanism for the removal of erythrocyte membrane patches containing removal molecules, thereby postponing the untimely elimination of otherwise healthy erythrocytes. Consequently, these same removal molecules mediate the rapid removal of erythrocyte-derived vesicles from the circulation. PMID- 18419624 TI - With the rapid development of new drugs to treat cancer and its effects, there will always be the risk factors as well as the therapeutic ones. PMID- 18419625 TI - Screening participation in individuals with a family history of colorectal cancer: a review. AB - Literature regarding screening behaviour in individuals with a family history of colorectal cancer was reviewed, in order to determine the prevalence of screening in this population and identify factors associated with screening participation. Four electronic databases were searched from 1994. Thirty papers met the inclusion criteria, including 3 community surveys, 13 studies on first-degree relatives of colorectal cancer patients, and 14 studies on genetic services for colorectal cancer risk assessment. Individuals with a family history of colorectal cancer, who have not received risk assessment, frequently have never had any form of screening for colorectal cancer. Uptake of endoscopic screening when offered to individuals identified as being at increased risk was generally high (often >60% participation). Having a medical recommendation to screen, a stronger family history and perceiving fewer barriers to screening were identified as predictors of screening behaviour. Existing data suggest that use of screening tests in individuals with a family history of colorectal cancer is variable, and our understanding of factors associated with screening behaviour is limited. A number of methodological problems in research to date were identified, and further research is needed in order to inform interventions to support sustained screening participation in this population. PMID- 18419626 TI - Involving people affected by cancer in research: a review of literature. AB - The purpose of the literature review was to find out why people affected by cancer have been involved in research; how they have been involved and the impact of their involvement. We used systematic methods to search for literature, applied inclusion and exclusion criteria, conducted a quality appraisal, selected relevant data from the included articles for analysis, and provided a narrative summary of these data. The literature shows that people affected by cancer, particularly women with breast cancer, have been involved in a range of research programmes, projects and initiatives especially in the USA, UK, Canada and Australia. Their involvement has impacted upon research design, accrual and response rates. There is increasing recognition of the need for an infrastructure, including formal recruitment procedures, training and mentoring, to support an agenda of involvement and a need to challenge the ethos of traditional research, which does not easily lend itself to this agenda. Further critique of the role of 'experiential knowledge' in research is required so that researchers and people affected by cancer can work in partnership. PMID- 18419627 TI - Diagnosed with breast cancer while on a family history screening programme: an exploratory qualitative study. AB - Mammographic screening is offered to many women under 50 in the UK who are at moderate or high risk of developing breast cancer because of their family history of the disease. Little is understood about the impact of screening on the emotional well-being of women with a family history of breast cancer. This qualitative study explores the value that women at increased risk placed on screening, both pre- and post-cancer diagnosis and the impact of the diagnosis. In-depth interviews were undertaken with 12 women, aged 35-50, diagnosed with breast cancer while on an annual mammographic screening programme. Women described the strong sense of reassurance gained from screening prior to diagnosis. This faith in screening was reinforced by early detection of their cancer. Reactions to diagnosis ranged from devastation to relief at having finally developed a long-expected condition. Despite their positive attitudes about screening, not all women wanted to continue with surveillance. For some, prophylactic mastectomy was preferable, to reduce future cancer risk and to alleviate anxieties about the detection of another cancer at each subsequent screen. This study illustrates the positive yet diverse attitudes towards mammographic screening in this group of women with a family history of breast cancer. PMID- 18419628 TI - Using undergraduate nursing students as mediators in a knowledge transfer programme for care for patients with advanced cancer. AB - Nursing today faces numerous challenges. Societal changes lead to reorganization of health care, changing workloads with sicker patients in hospital and home care, and limited economic resources. The increasing and changing nature of knowledge needed for expert care provision challenges nurses to continually update their competencies. These are issues demanding proactive and dynamic changes in the way nurses conceive their mandates and practice. The aim of the action-research project presented here was to foster improved quality of care for patients with advanced cancer through collaborative endeavours integrating cancer nursing clinical practice, research and education in a knowledge exchange programme. The programme was based on input about caregiving needs from multi professional staff caring for patients with advanced cancer in a variety of healthcare settings. Undergraduate baccalaureate nursing students were then engaged in literature studies to help address these needs. Results of the studies were communicated back to the involved clinicians in a variety of ways. In this paper, we discuss what we have experienced as opportunities and obstacles in conducting the project, based on our reflections and external evaluations. This is linked to a broader discussion of ways of integrating cancer nursing research, education and practice. PMID- 18419629 TI - Survival trends in osteosarcoma of humerus. AB - Management of osteosarcoma has evolved during the last 50 years. A review of 53 cases of osteosarcoma affecting the humerus included in the Scottish Bone Tumour registry from 1950 to 2000 is presented. Survival trends and the various factors influencing survival during this period are analysed. Patients were chronologically divided into two cohorts, one presenting before 1975 (18 patients) and the other presenting after 1975 (35 patients). The second half of the study period (1975 onwards) was characterized by availability of new treatment modalities like limb salvage and chemotherapy. A distinct improvement in survival was noted during the second half of the study (35% 5 years survival post 1975 vs. 18% pre 1975, P = 0.007). Survival and metastasis appeared to depend on multiple factors. Chemotherapy in isolation did not significantly affect survival. Limb salvage surgery did not have an adverse effect on survival rates. This improvement in survival could be due to a change in the natural history of the disease along with advances in diagnostics and therapeutics. PMID- 18419630 TI - Effects of fluconazole in the prophylaxis of oropharyngeal candidiasis in patients undergoing radiotherapy for head and neck tumour: results from a double blind placebo-controlled trial. AB - Fluconazole is recommended in the prophylaxis of oropharyngeal candidiasis (OPC) in patients undergoing radiotherapy for head-neck tumours; however, the actual effectiveness of fluconazole in this setting remains unclear. Adult patients with cervico-cephalic carcinoma submitted to radical or adjuvant radiotherapy were randomized to 100 mg fluconazole (n = 138) or matched placebo (n = 132) oral suspension once daily from the sixth session of radiotherapy up to the end of treatment. The final analysis of the investigation showed a higher rate of the OPC outbreak-free survival in the fluconazole compared with placebo (P = 0.008 in the log-rank test). The mean time (95% CI) to OPC outbreak was 56 (53-59) days in the fluconazole group and 47 (43-51) days with placebo. The mean duration of radiotherapy was 43.5 and 39.9 days, respectively in the two groups (P = 0.027). Adverse effects were reported in 70.3% of patients in the fluconazole group and in 67.4% with placebo. The results showed prophylaxis with fluconazole given in irradiated patients with head-neck tumours significantly reduces the rate and the time to development of OPC compared with placebo. PMID- 18419631 TI - Awareness of breast and cervical cancer risk factors and screening behaviours among nurses in rural region of Turkey. AB - Breast and cervical cancer are the most common causes of cancer mortality among women worldwide, but actually they are largely preventable diseases. There is limited data on breast and cervical cancer knowledge, screening practices and attitudes of nurses in Turkey. A self-administered questionnaire was used to investigate the knowledge and attitude of nurses on risk factors of the breast and cervical cancer as well as screening programmes such as breast self examination (BSE), clinical breast examination, mammography (MMG) and papanicolaou (pap) smear test. In total, 125 out of 160 nurses participated in the study (overall response rate was 80.6%). The risk factors and symptoms of breast cancer was generally well known, except for early menarche (23.2%) and late menopause (28.8%). For cervical cancer, the correct risk factors mostly indicated by the nurses were early age at first sexual intercourse (56%), smoking (76%), multiple sexual partners (71.2%). As for screening methods, it was believed that BSE was a beneficial method to identify the early breast changes (84.8%) and MMG was able to detect the cancer without a palpable mass (57.6%). Little was known about the fact that women should begin cervical cancer screening approximately 3 years after the onset of sexual intercourse (23.2%) and if repeated pap smear test were normal, it could be done every 2-3 years. Most of the nurses considered that MMG decreases the mortality in breast cancer (65.6%) and also believed that pap smear test decreases the mortality in cervical cancer (75.2%). Despite high level of knowledge of breast cancer risk factors, symptoms and screening methods, inadequate knowledge of cervical cancer screening method were found among nurses. PMID- 18419632 TI - Childhood leukaemia: experiences of children and attitudes of parents on dental care. AB - Parental perceptions in the importance of dental care and preferences with regard to its provision while profiling the level of dental health knowledge of parents of leukaemic children were elicited. The setting was the Paediatric Dental Care Unit located in Medical Faculty. Data were collected by means of a structured interview, employing a questionnaire. Level of knowledge on both dental facts and preventive dentistry of the participants was insufficient. Major source of dental care was the resident paediatric dentist both in prior to (78.2%) and following (100%) diagnosis. Tooth extraction (17.6%) was the only treatment provided prior to diagnosis. Following diagnosis, 60 (69%) of these children had received operative dental treatment. The source of preventive advice was inconsistent. Parents appeared to place a high level of importance on their children's dental care and the preference for this to be provided within the hospitals in which the child has been treated. There is clearly a need to establish dental care units in hospitals in which treatment of childhood malignancy is provided. The provision for the future should be the continuous education of dentists, physicians and nurses who work in hospitals and public health services. PMID- 18419633 TI - Cancer patients' view on rehabilitation and quality of life: a programme audit. AB - An existing 12-week cancer rehabilitation group programme, combining physical training and psychosocial sessions, was recently tested in a quantitative longitudinal cohort study (n = 658) among adult patients who have problems coping with cancer. Results showed improvements in all quality of life domains. The objective of the present study was to explore patients' perspectives on how and to what extent the rehabilitation programme had met their needs in order to optimize the programme. Patients who participated in the programme no longer than 5 years ago were interviewed in three focus groups (n = 23). Participants saw rehabilitation as provided by the programme as an important steppingstone in their ongoing recovery process and identified the integrated physical training, psychosocial sessions and patient-to-patient contacts as a powerful and supportive combination. Recommendations for improvements to the programme mainly concerned the psychosocial module. Information needs varied strongly among participants and, in this respect, it was hard to satisfy all of them. Participants suggested that the programme should focus more on developing skills to cope with cancer, its consequences and emerging problems after rehabilitation. In conclusion, participants valued the group-based multi-modal cancer rehabilitation programme and recommended to focus more on developing coping skills. PMID- 18419634 TI - Seating and pressure support needs of people with cancer in the cervix or rectum: a case series on the clinical usefulness of pressure mapping assessment. AB - Pressure sores present a challenge to people with rectal and cervical cancers due to weight loss, compromised nutrition, chemotherapy and radiotherapy. Affected individuals often experience intense pain on sitting, and prefer to spend the majority of their time lying down. This pilot study, employing a case study design, investigated the pressure care needs of such persons using pressure mapping, a technology designed to measure pressures at the seating interface. Four participants were mapped on a selection of five cushions, three of which were developed to specifically reduce midline posterior pressure/pain relief. Participants rated the cushions for comfort. Findings demonstrated an inconsistent relationship between interface pressures and perceived cushion comfort, suggesting that patient-rated comfort is a poor indicator of high interface pressures. The specialized cushions did not always meet the needs of this target population and no one cushion suited all. This study demonstrated the precarious clinical balance needed between comfort and achieving optimal pressure reduction in cushion prescription for this client group, and suggested that comfort was more important than pressure reduction in terms of their seating needs. PMID- 18419635 TI - Association of plasma methotrexate, neutropenia, hepatic dysfunction, nausea/vomiting and oral mucositis in children with cancer. AB - Oral mucositis is a major toxicity associated with high-dose methotrexate (HD MTX) therapy in the treatment of children with acute lymphoblastic leukaemia and osteosarcoma. This pilot matched case-control study investigated the associations between plasma concentration of MTX at 42 (p-MTX(42h)) and 66 (p-MTX(66h)) h, absolute neutrophil count (ANC) < or = or >1.0 x 10(9)/l, serum transaminases (ASAT/ALAT) < or > or =58 U/l, WHO < or > or =grade 2 nausea/vomiting and WHO < or > or =grade 2 oral mucositis. In this study, 11 children with WHO > or =grade 2 oral mucositis were compared with 17 control children matched for age, diagnosis and MTX-dosage. The results indicated that children with p-MTX(42h) > or = 1.0 micromol/l had an odds ratio (OR) of 4.3 of developing oral mucositis when compared with the referent group of children who had p-MTX(42h) < 1.0 micromol/l. Children with p-MTX(66h) >= 0.2 micromol/l had an OR of 8.2 of developing oral mucositis when compared with the referent group of children who had p-MTX(66h) < 0.2 micromol/l. Children with ANC < or = 1.0 x 10(9)/l had an OR of 1.2 of developing oral mucositis when compared with the referent group of children who had ANC > 1.0 x 10(9)/l. In comparison with the referent group of children, who had <58 U/l ASAT/ALAT, those with ASAT/ALAT > or = 58 U/l had an OR of 1.2 of developing oral mucositis. Finally, children with WHO grade > or =2 nausea/vomiting had an elevated risk of developing oral mucositis when compared with the referent group of children who had WHO grade <2 nausea/vomiting (OR = 8.7). In conclusion, the results in this preliminary study provide support for the hypothesis that the risk of oral mucositis is associated with the plasma MTX concentration at 66 h and the level of nausea/vomiting. PMID- 18419636 TI - An unusually early age of presentation of Phylloides tumour in Eastern Nigeria. PMID- 18419637 TI - Impact of non-alcoholic fatty liver disease on accelerated metabolic complications. AB - Insulin resistance is the basis of both non-alcoholic fatty liver disease (NAFLD) and metabolic syndrome (MetS), the two conditions are often found in the same individual. The mortality of patients with NAFLD is significantly higher than that among the general population and cardiovascular risk may compete with liver related risk in dictating the final outcome. Recent prospective studies have reported that NAFLD is associated with an increased incidence of MetS and type 2 diabetes mellitus, independent of obesity and other components of MetS. Thus, NAFLD may not only be a liver disease but also an early mediator of type 2 diabetes mellitus and MetS. The biological mechanisms by which NAFLD contributes to a higher risk of developing metabolic disorders are not fully understood. However, the fatty liver could contribute in the same way as visceral adipose tissue to insulin resistance, systemic inflammation and oxidative stress, while the decreased serum adiponectin concentrations might also be part of the mechanism. In contemporary clinical practice, it has become mandatory to evaluate the metabolic risk factors in NAFLD patients and to consider careful surveillance and aggressive treatment, not only of the resultant liver disease, but also of the possible underlying metabolic and vascular complications. Future studies might address the question whether earlier adjustment to a more efficient lifestyle or a pharmacological treatment that mobilizes fat out of the liver could reduce these risks. PMID- 18419638 TI - Advances in gene chip technique in Barrett's metaplasia and adenocarcinoma. AB - Gene chip methods are applied to the study of gene expression. The differentially expressed genes in different specimens may be detected with parallel analysis by gene chip, which has greatly improved the traditional experiments in that only a single or several gene expressions need to be observed for each test, thereby speeding up the identification of differentially expressed genes and the construction of differential expression profiles. Many studies have applied this technology for Barrett's metaplasia and adenocarcinoma, and identified a number of candidate genes useful as biomarkers in cancer staging, prediction of recurrence, prognosis and treatment selection. This review described the gene expression profile and molecular changes related to Barrett's metaplasia and adenocarcinoma of the esophagus, with emphasis on its prognostic value and possibilities for targeted therapy in a clinical setting. PMID- 18419639 TI - Endoscopic clipping. AB - Endoscopic clips are relatively new devices that have been shown to be effective for the control of acute gastrointestinal hemorrhage. Various different models are available and offer simplicity of use with relatively few complications. Recently, endoscopic clips have been used for a variety of non-hemorrhagic conditions. In this article we review the literature and present current thinking about the indications, efficacy and safety of these devices. PMID- 18419640 TI - CYP2C9 polymorphism in non-steroidal anti-inflammatory drugs-induced gastropathy. AB - OBJECTIVE: Non-steroidal anti-inflammatory drugs (NSAID) induce gastroduodenal mucosal injury and are metabolized by cytochrome P450 2C9 (CYP2C9). It is postulated that CYP2C9 genotype is associated with NSAID-induced gastropathy. This study aims to determine whether individuals with a CYP2C9 allele mutation are susceptible to NSAID-induced gastropathy. METHODS: A total of 109 patients diagnosed as having rheumatic diseases and taking NSAID were appraised as having gastropathy by endoscopy, stool occult blood test and questionnaire two weeks after entering the study. Their peripheral blood was analyzed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). RESULTS: A total of 47.7% gastropathy (33% erosions, 14.7% ulcers, 2.75% ulcer bleeding) and 56% dyspeptic symptoms were presented. Only one CYP2C9*2 heterozygote (*1/*2) was found in the group with gastropathy and two variant alleles (CYP2C9*2 and CYP2C9* 3) could not be found in the group without gastropathy. There was no significant difference in both CYP2C9 genotype (0.96%vs 0%) and CYP2C9 variant allele frequency (1.92%vs 0%) between patients with and without gastropathy. CONCLUSION: These results confirm the high prevalence of NSAID-induced gastropathy but do not support the postulation that CYP2C9*2 and CYP2C9*3 contribute to the development of NSAID-induced gastropathy. This may be due to the low frequency of the two alleles in the population studied. PMID- 18419641 TI - Efficacy and safety of azathioprine maintenance therapy in a group of Crohn's disease patients in China. AB - OBJECTIVE: To evaluate the efficacy and the safety of long-term azathioprine maintenance therapy in a group of Chinese patients with Crohn's disease. METHODS: The efficacy of azathioprine (2.0 mg/kg/day) in controlling the disease relapse in 13 patients with Crohn's disease following clinical remission by prednisone or surgery were investigated. The Crohn's disease activity index (CDAI) and the Harvey-Bradshaw index, the reduction of steroid dosage and side-effects for an average of 18 months follow up were analyzed. RESULTS: Azathioprine was effective in controlling the disease relapse in 10 (by CDAI scores) or 11 (by Harvey Bradshaw index) of 13 patients (76.9% and 84.6%, respectively) for at least 6 months. Azathioprine was not discontinued in a patient who experienced a temporary and mild elevation of aminotransferases 14 months after the initiation of therapy. However one patient who was co-administered with azathioprine and mesalamine (Pentasa) developed an episode of bone marrow suppression that ultimately required the withdrawal of both medications. CONCLUSION: Azathioprine is an effective agent which controls the relapse of Crohn's disease in most patients. Long-term remission can be achieved. Side-effects, including severe leukopenia, myelo-suppression and the mild elevation of hepatic enzymes, may occur in a small number of patients. PMID- 18419642 TI - A homozygous mutation in a Chinese man with Crigler-Najjar syndrome type II and a family genetic analysis. AB - OBJECTIVE: To investigate the family genetic background of a 22-year-old man with Crigler-Najjar syndrome type II (CN-II). METHODS: After the proband (patient) with CN-II was diagnosed by liver function tests, a low calorie intake test and an oral phenobarbital enzyme-induction trial, blood samples were collected from 11 family members for identifying DNA gene groups. Exons 1-5 of the UGT1A1 gene were amplified by polymerase chain reaction (PCR) and mutations of the UGT1A1 gene were screened by a direct DNA sequencing. RESULTS: The serum unconjugated bilirubin increased in the proband from 156.4 micromol/L to 243.5 micromol/L after he started a low calorie intake, and it decreased to 51.8 micromol/L within a month of taking oral phenobarbital daily. Both functional tests and ultrasonographic images of the liver were normal except for the unconjugated hyperbilirubinemia. A missense mutation of Tyr486Asp at exon 5 in the UGT1A1 gene and a homozygous mutation were confirmed in the proband. Heterozygous mutations were found in his parents, his younger sister and three great-uncles, while no mutation of the UGT1A1 gene was detected in the remaining four family members. CONCLUSION: A missense mutation of Tyr486Asp is considered to be the cause of the CN-II in this patient. It is a recessive trait that is autosomally inherited in this family. No influence of the mutation was found on the response elements for phenobarbital in the promoter region. PMID- 18419643 TI - High prevalence of hepatitis C virus infection in the largest province of Pakistan. AB - OBJECTIVE: To estimate the prevalence and spectrum of hepatitis C virus (HCV) infection in the general population of Pakistan. METHODS: A total of 6817 blood samples were collected randomly from apparently healthy people in the Punjab, Pakistan from March 1999 to April 2001 and September 2006 to August 2007. Detailed socioeconomic information for each participant was recorded. All the samples were tested for anti-HCV antibodies and all seropositive samples were further tested for HCV RNA by polymerase chain reaction (PCR). RESULTS: Of the total 6817 serum samples tested, 998 (14.63%) were positive for anti-HCV antibodies. HCV RNA PCR was detected in 494 (49.50%) anti-HCV-positive samples. The prevalence of anti-HCV antibodies were significantly higher in males (15.09%) than in females (12.3%) (P < 0.009). A significant difference was also noted in the anti-HCV prevalence rate among different age groups tested (P < 0.01). In a multivariate logistic regression analysis, injected drug use (adjusted OR 6.6 [95%CI 4.1-9.9]), blood transfusion (adjusted OR 5.9 [95%CI 2.9-12.3]), pricked with a needle (adjusted OR 2.2 [95%CI 1.6-3.1]), re-use of syringes (adjusted OR 1.7 [95%CI 0.8-3.6]) and being over 35 years old (adjusted OR 1.3 [95%CI 0.9 1.9]) were independent risk factors for HCV infection. CONCLUSION: The study showed a high seroprevalence of anti-HCV antibodies in a general and apparently healthy population of the Punjab province of Pakistan. Drug injection, blood transfusion and needle stuck were the factors most strongly associated with HCV infection. PMID- 18419644 TI - Synchronous early gastric cancer and gastrointestinal stromal tumor in the stomach of a patient with idiopathic thrombocytopenic purpura. AB - The synchronous occurrence of gastrointestinal stromal tumour (GIST) in the stomach and early gastric cancer is uncommon, with only a few previous reports. In particular, the collision of GIST and early gastric cancer in a patient with idiopathic thrombocytopenic purpura (ITP) has never been reported. We present the case of a 78-year-old male patient with ITP who was diagnosed with a synchronous development of GIST and early gastric cancer of the stomach. He underwent an elective subtotal gastrectomy with splenectomy. We discuss whether the development of GIST in the stomach in concert with early gastric cancer is an incidental coexistence or involve the same carcinogenic agents. Furthermore, it is not known whether or not such a situation is connected with ITP. To our knowledge this is the second report of a small GIST concomitant with an early gastric cancer and the first one in a patient with ITP. PMID- 18419645 TI - Guidelines for the diagnosis and treatment of nonalcoholic fatty liver diseases. PMID- 18419646 TI - Guidelines for the diagnosis and treatment of alcoholic liver disease. PMID- 18419647 TI - Estimating consumer exposure to PFOS and PFOA. AB - Perfluorinated compounds have been used for more than 50 years as process aids, surfactants, and for surface protection. This study is a comprehensive assessment of consumer exposure to perfluorooctane sulfonate (PFOS) and perfluorooctanoic acid (PFOA) from a variety of environmental and product-related sources. To identify relevant pathways leading to consumer exposure to PFOS and PFOA a scenario-based approach has been applied. Scenarios represent realistic situations where age- and gender-specific exposure occurs in the everyday life of consumers. We find that North American and European consumers are likely to experience ubiquitous and long-term uptake doses of PFOS and PFOA in the range of 3 to 220 ng per kg body weight per day (ng/kg(bw)/day) and 1 to 130 ng/kg(bw)/day, respectively. The greatest portion of the chronic exposure to PFOS and PFOA is likely to result from the intake of contaminated foods, including drinking water. Consumer products cause a minor portion of the consumer exposure to PFOS and PFOA. Of these, it is mainly impregnation sprays, treated carpets in homes, and coated food contact materials that may lead to consumer exposure to PFOS and PFOA. Children tend to experience higher total uptake doses (on a body weight basis) than teenagers and adults because of higher relative uptake via food consumption and hand-to-mouth transfer of chemical from treated carpets and ingestion of dust. The uptake estimates based on scenarios are within the range of values derived from blood serum data by applying a one-compartment pharmacokinetic model. PMID- 18419648 TI - Applicability of a neuroprobabilistic integral risk index for the environmental management of polluted areas: a case study. AB - Recently, we developed a GIS-Integrated Integral Risk Index (IRI) to assess human health risks in areas with presence of environmental pollutants. Contaminants were previously ranked by applying a self-organizing map (SOM) to their characteristics of persistence, bioaccumulation, and toxicity in order to obtain the Hazard Index (HI). In the present study, the original IRI was substantially improved by allowing the entrance of probabilistic data. A neuroprobabilistic HI was developed by combining SOM and Monte Carlo analysis. In general terms, the deterministic and probabilistic HIs followed a similar pattern: polychlorinated biphenyls (PCBs) and light polycyclic aromatic hydrocarbons (PAHs) were the pollutants showing the highest and lowest values of HI, respectively. However, the bioaccumulation value of heavy metals notably increased after considering a probability density function to explain the bioaccumulation factor. To check its applicability, a case study was investigated. The probabilistic integral risk was calculated in the chemical/petrochemical industrial area of Tarragona (Catalonia, Spain), where an environmental program has been carried out since 2002. The risk change between 2002 and 2005 was evaluated on the basis of probabilistic data of the levels of various pollutants in soils. The results indicated that the risk of the chemicals under study did not follow a homogeneous tendency. However, the current levels of pollution do not mean a relevant source of health risks for the local population. Moreover, the neuroprobabilistic HI seems to be an adequate tool to be taken into account in risk assessment processes. PMID- 18419649 TI - Modeling exposure to air pollution from the WTC disaster based on reports of perceived air pollution. AB - We examined the utility of a newly developed perceived air pollution (PAP) scale and of a modeled air pollution (MAP) scale derived from it for predicting previously observed birth outcomes of pregnant women enrolled following September 11, 2001. Women reported their home and work locations in the four weeks after September 11, 2001 and the PAP at each site on a four-point scale designed for this purpose. Locations were geocoded and their distance from the World Trade Center (WTC) site determined. PAP values were used to develop a model of air pollution for a 20-mile radius from the WTC site. MAP values were assigned to each geocoded location. We examined the relationship of PAP and MAP values to maternal characteristics and to distance of home and work sites from the WTC site. Both PAP and MAP values were highly correlated with distance from the WTC. Maternal characteristics that were associated with PAP values reported for home or work sites (race, demoralization, material hardship, first trimester on September 11) were not associated with modeled MAP values. Relationships of several birth outcomes to proximity to the WTC, which we previously reported using this data set, were also seen when MAP values were used as the measure of exposure, instead of proximity. MAP developed from reports of PAP may be useful to identify high-risk areas and predict health outcomes when there are multiple sources of pollution and a "distance from source" analysis is impossible. PMID- 18419650 TI - Rate of foot-and-mouth disease virus transmission by carriers quantified from experimental data. AB - Upon infection with foot-and-mouth disease virus (FMDV) a considerable number of animals become carriers of the virus. These carriers are considered to be a risk for new outbreaks, but the rate at which these animals can transmit the infection has not been quantified. An analysis was carried out using data from previously published experiments in order to quantify the transmission rate parameter beta of FMDV infection from carriers to susceptible animals. The parameter beta was estimated at 0.0256 (likelihood-based confidence interval: 0.008-0.059) infections per carrier per month. Moreover, analysis of published experimental data indicates that the proportion of FMDV carriers decreases at a rate of 0.115 per month. Both parameters obtained from this study are useful for quantitative risk analyses of the trade of animals from FMDV-infected areas or the lifting of vaccination programs. PMID- 18419651 TI - European public perceptions of food risk: cross-national and methodological comparisons. AB - This article presents a comparative study of public perceptions of food risk across 25 European member states. A secondary data analysis is conducted on a Eurobarometer survey fielded to nationally representative samples in 2005. The survey included closed questions as well as free associations to map risk perceptions. Taking a quantitative approach, we find that people in a majority of European countries express similar levels of concern about food risks. However, outside this majority a North-South divide is evident, with the Northern countries worrying less than the Southern countries. Multilevel modeling shows that cross-national differences in individual respondents' level of worry are in part attributable to shared country effects and to generalized risk sensitivity about a range of personal risks. On the underlying structure of food risk concerns, factor analysis points to three dimensions described by groupings of risks related to adulteration and contamination, health effects, and production and hygiene. A qualitative analysis of respondents' free associations about problems and risks with food identifies three major themes that are consistent with the quantitative results. However, the free associations also point toward greater cross-national diversity and to striking variations in the range and importance of food risks. Overall, the picture is of a public that frames food risks in a wider context of beliefs about the links between diet and health. We conclude with some implications for research on food risk perceptions in particular and risk perception studies in general. PMID- 18419652 TI - Using probabilistic terrorism risk modeling for regulatory benefit-cost analysis: application to the Western hemisphere travel initiative in the land environment. AB - This article presents a framework for using probabilistic terrorism risk modeling in regulatory analysis. We demonstrate the framework with an example application involving a regulation under consideration, the Western Hemisphere Travel Initiative for the Land Environment, (WHTI-L). First, we estimate annualized loss from terrorist attacks with the Risk Management Solutions (RMS) Probabilistic Terrorism Model. We then estimate the critical risk reduction, which is the risk reducing effectiveness of WHTI-L needed for its benefit, in terms of reduced terrorism loss in the United States, to exceed its cost. Our analysis indicates that the critical risk reduction depends strongly not only on uncertainties in the terrorism risk level, but also on uncertainty in the cost of regulation and how casualties are monetized. For a terrorism risk level based on the RMS standard risk estimate, the baseline regulatory cost estimate for WHTI-L, and a range of casualty cost estimates based on the willingness-to-pay approach, our estimate for the expected annualized loss from terrorism ranges from $2.7 billion to $5.2 billion. For this range in annualized loss, the critical risk reduction for WHTI-L ranges from 7% to 13%. Basing results on a lower risk level that results in halving the annualized terrorism loss would double the critical risk reduction (14-26%), and basing the results on a higher risk level that results in a doubling of the annualized terrorism loss would cut the critical risk reduction in half (3.5-6.6%). Ideally, decisions about terrorism security regulations and policies would be informed by true benefit-cost analyses in which the estimated benefits are compared to costs. Such analyses for terrorism security efforts face substantial impediments stemming from the great uncertainty in the terrorist threat and the very low recurrence interval for large attacks. Several approaches can be used to estimate how a terrorism security program or regulation reduces the distribution of risks it is intended to manage. But, continued research to develop additional tools and data is necessary to support application of these approaches. These include refinement of models and simulations, engagement of subject matter experts, implementation of program evaluation, and estimating the costs of casualties from terrorism events. PMID- 18419653 TI - Risk-based analysis of the Danish pork Salmonella program: past and future. AB - The Danish pork Salmonella control program was initiated in 1993 in response to a prominent pork-related outbreak in Copenhagen. It involved improved efforts at slaughter hygiene (postharvest) and on-farm (preharvest) surveillance and control. After 10 years, 95 million Euros, significant reductions in seropositive herds, Salmonella positive carcasses, and pork-attributable human cases (PAHC), questions have arisen about how best to continue this program. The objective of this study was to provide some analysis and information to address these questions. The methods used include a computer simulation model constructed of a series of Excel workbooks, one for each simulated year and scenario (http://www.ifss.iastate/DanSalmRisk). Each workbook has three modules representing the key processes affecting risk: seropositive pigs leaving the farm (Production), carcass contamination after slaughter (Slaughter), and PAHC of Salmonella (Attribution). Parameter estimates are derived from an extensive farm to-fork database collected by industry and government and managed by the Danish Zoonosis Centre (http://www.food.dtu.dk). Retrospective (1994-2003) and prospective (2004-2013) simulations were evaluated. The retrospective simulations showed that, except for the first few years (1994-1998), the on-farm program had minimal impact in reducing the number of positive carcasses and PAHC. Most of the reductions in PAHC up to 2003 were, according to this analysis, due to various improvements in abattoir processes. Prospective simulations showed that minimal reductions in human health risk (PAHC) could be achieved with on-farm programs alone. Carcass decontamination was shown as the most effective means of reducing human risk, reducing PAHC to about 10% of the simulated 2004 level. PMID- 18419654 TI - Optimal stopping analysis of a radiation detection system to protect cities from a nuclear terrorist attack. AB - We formulate and analyze an optimal stopping problem concerning a terrorist who is attempting to drive a nuclear or radiological weapon toward a target in a city center. In our model, the terrorist needs to travel through a two-dimensional lattice containing imperfect radiation sensors at some of the nodes, and decides at each node whether to detonate the bomb or proceed. We consider five different scenarios containing various informational structures and two different sensor array topologies: the sensors are placed randomly or they form an outer wall around the periphery of the city. We find that sensors can act as a deterrent in some cases, and that the government prefers the outer wall topology unless the sensors have a very low detection probability and the budget is tight (so that they are sparsely deployed). PMID- 18419655 TI - Communicating the risks of a new, emerging pathogen: the case of Cryptococcus gattii. AB - The news media are an important channel for communicating public health messages, providing important information about geographic extent, symptoms, and precautionary measures. This function may be particularly important in the case of new and emerging infectious diseases, which are unfamiliar to both the public and health professionals. We conducted a content analysis of the coverage in six Canadian newspapers of two emerging infectious pathogens in British Columbia (BC), West Nile virus and Cryptococcus gattii (C. gattii), between 2001 and 2006. C. gattii was first identified in the Canadian Pacific Northwest in 1999, having previously been restricted to tropical and subtropical areas. By summer 2006, C. gattii had killed at least eight people and was responsible for over 150 hospitalizations in the province of BC, as well as significant morbidity and mortality among companion animals. West Nile virus reached Canada in 2001 and has since caused significant morbidity and mortality. It has not, to date, reached BC. Despite the much higher incidence of disease and death caused by C. gattii in BC, West Nile virus received five times more news coverage in the British Columbian and national newspapers examined. Surprisingly, the local newspaper closest to the center of the British Columbia C. gattii outbreak had a much lower proportion of coverage of the disease than other papers in the province. The article discusses possible explanations for these findings, and the implications for public health communication. PMID- 18419656 TI - Hazard perception, risk perception, and the need for decontamination by residents exposed to soil pollution: the role of sustainability and the limits of expert knowledge. AB - This case study examines the hazard and risk perception and the need for decontamination according to people exposed to soil pollution. Using an ecological-symbolic approach (ESA), a multidisciplinary model is developed that draws upon psychological and sociological perspectives on risk perception and includes ecological variables by using data from experts' risk assessments. The results show that hazard perception is best predicted by objective knowledge, subjective knowledge, estimated knowledge of experts, and the assessed risks. However, experts' risk assessments induce an increase in hazard perception only when residents know the urgency of decontamination. Risk perception is best predicted by trust in the risk management. Additionally, need for decontamination relates to hazard perception, risk perception, estimated knowledge of experts, and thoughts about sustainability. In contrast to the knowledge deficit model, objective and subjective knowledge did not significantly relate to risk perception and need for decontamination. The results suggest that residents can make a distinction between hazards in terms of the seriousness of contamination on the one hand, and human health risks on the other hand. Moreover, next to the importance of social determinants of environmental risk perception, this study shows that the output of experts' risk assessments-or the objective risks-can create a hazard awareness rather than an alarming risk consciousness, despite residents' distrust of scientific knowledge. PMID- 18419657 TI - Assessing the multidimensionality of coastal erosion risks: public participation and multicriteria analysis in a Mediterranean coastal system. AB - The complex and multidimensional nature of coastal erosion risks makes it necessary to move away from single-perspective assessment and management methods that have conventionally predominated in coastal management. This article explores the suitability of participatory multicriteria analysis (MCA) for improving the integration of diverse expertises and values and enhancing the social-ecological robustness of the processes that lead to the definition of relevant policy options to deal with those risks. We test this approach in the Mediterranean coastal locality of Lido de Sete in France. Results show that the more adaptive alternatives such as "retreating the shoreline" were preferred by our selected stakeholders to those corresponding to "protecting the shoreline" and the business as usual proposals traditionally put forward by experts and policymakers on these matters. Participative MCA contributed to represent coastal multidimensionality, elicit and integrate different views and preferences, facilitated knowledge exchange, and allowed highlighting existing uncertainties. PMID- 18419658 TI - Insurance against climate change and flooding in the Netherlands: present, future, and comparison with other countries. AB - Climate change is projected to cause severe economic losses, which has the potential to affect the insurance sector and public compensation schemes considerably. This article discusses the role insurance can play in adapting to climate change impacts. The particular focus is on the Dutch insurance sector, in view of the Netherlands being extremely vulnerable to climate change impacts. The usefulness of private insurance as an adaptation instrument to increased flood risks is examined, which is currently unavailable in the Netherlands. It is questioned whether the currently dominant role of the Dutch government in providing damage relief is justified from an economic efficiency perspective. Characteristics of flood insurance arrangements in the Netherlands, the United Kingdom, Germany, and France are compared in order to identify possible future directions for arrangements in the Netherlands. It is argued that social welfare improves when insurance companies take responsibility for part of the risks associated with climate change. PMID- 18419659 TI - Human salmonellosis: estimation of dose-illness from outbreak data. AB - The quantification of the relationship between the amount of microbial organisms ingested and a specific outcome such as infection, illness, or mortality is a key aspect of quantitative risk assessment. A main problem in determining such dose response models is the availability of appropriate data. Human feeding trials have been criticized because only young healthy volunteers are selected to participate and low doses, as often occurring in real life, are typically not considered. Epidemiological outbreak data are considered to be more valuable, but are more subject to data uncertainty. In this article, we model the dose-illness relationship based on data of 20 Salmonella outbreaks, as discussed by the World Health Organization. In particular, we model the dose-illness relationship using generalized linear mixed models and fractional polynomials of dose. The fractional polynomial models are modified to satisfy the properties of different types of dose-illness models as proposed by Teunis et al. Within these models, differences in host susceptibility (susceptible versus normal population) are modeled as fixed effects whereas differences in serovar type and food matrix are modeled as random effects. In addition, two bootstrap procedures are presented. A first procedure accounts for stochastic variability whereas a second procedure accounts for both stochastic variability and data uncertainty. The analyses indicate that the susceptible population has a higher probability of illness at low dose levels when the combination pathogen-food matrix is extremely virulent and at high dose levels when the combination is less virulent. Furthermore, the analyses suggest that immunity exists in the normal population but not in the susceptible population. PMID- 18419660 TI - Avian influenza in Vietnam: chicken-hearted consumers? AB - This study, based on quantitative and qualitative surveys conducted from July 2004 to September 2005, examines the perceptions of Hanoi consumers and their reactions to the Avian Influenza epizootic (H5N1). Hanoi consumers clearly link the risk of human contamination by the virus to the preparation and ingestion of poultry. During the first crisis, consumers reacted quickly and intensely (74% of them had already stopped eating poultry in January 2004). Nevertheless, once the crisis abated, they quickly resumed their consumption of poultry. This behavior corresponds to the pattern described by empirical studies of other crises, such as BSE. What is more surprising is the speed with which the different steps of this common pattern succeeded one another. It may be explained by a rapid decrease in risk anxiety. A logit model shows that, soon after the beginning of the crisis, AI risk anxiety was tempered by confidence in the information and recommendations issued by the government concerning AI and, in the long term, by a high perceived self-efficiency to deal with AI. Indeed, not only has poultry consumption been affected in terms of the quantity consumed, but alternative ways of selecting and preparing poultry have also been adopted as anti-risk practices. Risk communication strategies should take this into account, and rely on a previous assessment of consumer practices adopted to deal with the risk. PMID- 18419661 TI - Significant factors of aviation insurance and risk management strategy: an empirical study of Taiwanese airline carriers. AB - Aviation insurance premiums have become a heavy burden for the airline industry since September 11, 2001. Although the industry must constantly balance its operations between profitability and safety, the reality is that airlines are in the business of making money. Therefore, their ability to reduce cost and manage risk is a key factor for success. Unlike past research, which used subjective judgment methods, this study applied quantitative historical data (1999-2000) and gray relation analysis to identify the primary factors influencing ratemaking for aviation insurance premiums. An empirical study of six airlines in Taiwan was conducted to determine these factors and to analyze the management strategies used to deal with them. Results showed that the loss experience and performance of individual airlines were the key elements associated with aviation insurance premiums paid by each airline. By identifying and understanding the primary factors influencing ratemaking for aviation insurance, airlines will better understand their relative operational strengths and weaknesses, and further help top management identify areas for further improvement. Knowledge of these factors combined with effective risk management strategies, may result in lower premiums and operating costs for airline companies. PMID- 18419662 TI - Farmer knowledge and risk analysis: postrelease evaluation of herbicide-tolerant canola in Western Canada. AB - The global controversy regarding the use of genetically modified (GM) crops has proved to be a challenge for "science-based" risk assessments. Although risk analysis incorporates societal perspectives in decision making over these crops, it is largely predicated on contrasts between "expert" and "lay" perspectives. The overall objective of this study is to explore the role for farmers' knowledge, and their decade-long experience with herbicide-tolerant (HT) canola, in the risk analysis of GM crops. From 2002 to 2003, data were collected using interviews (n= 15) and mail surveys (n= 370) with farmers from Manitoba and across Canada. The main benefits associated with HT canola were management oriented and included easier weed control, herbicide rotation, and better weed control, whereas the main risks were more diverse and included market harm, technology use agreements (TUAs), and increased seed costs. Benefits and risks were inversely related, and the salient factor influencing risk was farmer experiences with HT canola volunteers, followed by small farm size and duration using HT canola. These HT volunteers were reported by 38% of farmers, from both internal (e.g., seedbank, farm machinery, etc.) and external (e.g., wind, seed contamination, etc.) sources, and were found to persist over time. Farmer knowledge is a reliable and rich source of information regarding the efficacy of HT crops, demonstrating that individual experiences are important to risk perception. The socioeconomic nature of most risks combined with the continuing "farm income crisis" in North America demonstrates the need for a more holistic and inclusive approach to risk assessment associated with HT crops and, indeed, with all new agricultural technology. PMID- 18419663 TI - Perspective: risk apportionment and disease intervention strategies. AB - The ultimate public health objective is the ability to predict and prevent disease, and not necessarily to identify an exhaustive list of potential disease risk factors. For any important public health outcome with multiple and potentially interrelated risk factors, an improved understanding of the contribution of individual and combinations of modifiable risk factors to the disease burden is essential for formulating an appropriate public health strategy. Partitioning techniques that divide the combined impact of multiple risk factors into exposure-specific components while taking into account the potential interrelations among those components, have been described in the epidemiological literature. In this article, we review and compare the available methods and options for such apportionment and apply them in a more general public health context as a method of selecting and prioritizing coronary heart disease (CHD) prevention strategies. PMID- 18419664 TI - A neural network-based method for risk factor analysis of West Nile virus. AB - There is a lack of knowledge about which risk factors are more important in West Nile virus (WNV) transmission and risk magnitude. A better understanding of the risk factors is of great help in developing effective new technologies and appropriate prevention strategies for WNV infection. A contribution analysis of all risk factors in WNV infection would identify those major risk factors. Based on the identified major risk factors, measures to control WNV proliferation could be directed toward those significant risk factors, thus improving the effectiveness and efficiency in developing WNV control and prevention strategies. Neural networks have many generally accepted advantages over conventional analytical techniques, for instance, ability to automatically learn the relationship between the inputs and outputs from training data, powerful generalization ability, and capability of handling nonlinear interactions. In this article, a neural network model was developed for analysis of risk factors in WNV infection. To reveal the relative contribution of the input variables, the neural network was trained using an algorithm called structural learning with forgetting. During the learning, weak neural connections are forced to fade away while a skeletal network with strong connections emerges. The significant risk factors can be identified by analyzing this skeletal network. The proposed approach is tested with the dead bird surveillance data in Ontario, Canada. The results demonstrate the effectiveness of the proposed approach. PMID- 18419665 TI - What's wrong with risk matrices? AB - Risk matrices-tables mapping "frequency" and "severity" ratings to corresponding risk priority levels-are popular in applications as diverse as terrorism risk analysis, highway construction project management, office building risk analysis, climate change risk management, and enterprise risk management (ERM). National and international standards (e.g., Military Standard 882C and AS/NZS 4360:1999) have stimulated adoption of risk matrices by many organizations and risk consultants. However, little research rigorously validates their performance in actually improving risk management decisions. This article examines some mathematical properties of risk matrices and shows that they have the following limitations. (a) Poor Resolution. Typical risk matrices can correctly and unambiguously compare only a small fraction (e.g., less than 10%) of randomly selected pairs of hazards. They can assign identical ratings to quantitatively very different risks ("range compression"). (b) Errors. Risk matrices can mistakenly assign higher qualitative ratings to quantitatively smaller risks. For risks with negatively correlated frequencies and severities, they can be "worse than useless," leading to worse-than-random decisions. (c) Suboptimal Resource Allocation. Effective allocation of resources to risk-reducing countermeasures cannot be based on the categories provided by risk matrices. (d) Ambiguous Inputs and Outputs. Categorizations of severity cannot be made objectively for uncertain consequences. Inputs to risk matrices (e.g., frequency and severity categorizations) and resulting outputs (i.e., risk ratings) require subjective interpretation, and different users may obtain opposite ratings of the same quantitative risks. These limitations suggest that risk matrices should be used with caution, and only with careful explanations of embedded judgments. PMID- 18419666 TI - Effects of risk representation and scope on willingness to pay for reduced risks: evidence from Tokyo Bay, Japan. AB - When applying the contingent valuation method (CVM) in risk reduction studies, some studies report that willingness to pay (WTP) is insensitive to the magnitude of risk reduction while other studies do not. On the other hand, social psychological research has shown that the affect heuristic biases judgments on probability, relative frequency, and risk. This article examines both magnitude (or scope) effect and affect heuristic (or representation) effect on WTP for risk reduction measures against tsunamis by introducing two experimental (i.e., absolute and relative) systems with four different representations to evaluate public behaviors in two different scenarios of risk reduction. Two common denominators (100 and 10,000) are introduced into absolute risk reduction representation (i.e., "of every 100 persons, from present 2 deaths to 1") to form different formats (i.e., "of every 10,000 persons from 200 to 100," and "of every 100,000 persons from 2,000 deaths to 1,000"). There is little evidence that WTP estimates are sufficiently sensitive to the magnitude of risk reduction, but relative risk reduction representations may be better than the absolute one given in CVM mail surveys when the risk is small. There is a statistically insignificant effect of risk reduction representations on respondent frequency, but mixed effects on the monetary values of WTP at the level of 0.05. The representation effect of absolute risk reduction on the WTP value varies with the common denominator. The larger the common denominator, the less the WTP to reduce the risk of tsunamis, and the significance probability is improved to less than the level of 0.05 when the common denominator becomes large enough. The findings suggest that improved methods are required for estimating the rates of tradeoff between fatality risk and other goods among consumers. PMID- 18419667 TI - Implied preference for seismic design level and earthquake insurance. AB - Seismic risk can be reduced by implementing newly developed seismic provisions in design codes. Furthermore, financial protection or enhanced utility and happiness for stakeholders could be gained through the purchase of earthquake insurance. If this is not so, there would be no market for such insurance. However, perceived benefit associated with insurance is not universally shared by stakeholders partly due to their diverse risk attitudes. This study investigates the implied seismic design preference with insurance options for decisionmakers of bounded rationality whose preferences could be adequately represented by the cumulative prospect theory (CPT). The investigation is focused on assessing the sensitivity of the implied seismic design preference with insurance options to model parameters of the CPT and to fair and unfair insurance arrangements. Numerical results suggest that human cognitive limitation and risk perception can affect the implied seismic design preference by the CPT significantly. The mandatory purchase of fair insurance will lead the implied seismic design preference to the optimum design level that is dictated by the minimum expected lifecycle cost rule. Unfair insurance decreases the expected gain as well as its associated variability, which is preferred by risk-averse decisionmakers. The obtained results of the implied preference for the combination of the seismic design level and insurance option suggest that property owners, financial institutions, and municipalities can take advantage of affordable insurance to establish successful seismic risk management strategies. PMID- 18419668 TI - Households' perceived personal risk and responses in a multihazard environment. AB - This study proposed and tested a multistage model of household response to three hazards-flood, hurricane, and toxic chemical release-in Harris County Texas. The model, which extends Lindell and Perry's (1992, 2004) Protective Action Decision Model, proposed a basic causal chain from hazard proximity through hazard experience and perceived personal risk to expectations of continued residence in the home and adoption of household hazard adjustments. Data from 321 households generally supported the model, but the mediating effects of hazard experience and perceived personal risk were partial rather than complete. In addition, the data suggested that four demographic variables-gender, age, income, and ethnicity affect the basic causal chain at different points. PMID- 18419669 TI - Quantitative risk assessment from farm to fork and beyond: a global Bayesian approach concerning food-borne diseases. AB - A novel approach to the quantitative assessment of food-borne risks is proposed. The basic idea is to use Bayesian techniques in two distinct steps: first by constructing a stochastic core model via a Bayesian network based on expert knowledge, and second, using the data available to improve this knowledge. Unlike the Monte Carlo simulation approach as commonly used in quantitative assessment of food-borne risks where data sets are used independently in each module, our consistent procedure incorporates information conveyed by data throughout the chain. It allows "back-calculation" in the food chain model, together with the use of data obtained "downstream" in the food chain. Moreover, the expert knowledge is introduced more simply and consistently than with classical statistical methods. Other advantages of this approach include the clear framework of an iterative learning process, considerable flexibility enabling the use of heterogeneous data, and a justified method to explore the effects of variability and uncertainty. As an illustration, we present an estimation of the probability of contracting a campylobacteriosis as a result of broiler contamination, from the standpoint of quantitative risk assessment. Although the model thus constructed is oversimplified, it clarifies the principles and properties of the method proposed, which demonstrates its ability to deal with quite complex situations and provides a useful basis for further discussions with different experts in the food chain. PMID- 18419671 TI - The potential role of soyfoods in weight and adiposity reduction: an evidence based review. AB - Evidence concerning the relationship between soyfoods and weight loss was reviewed. Detailed searches of PubMed and Web of Science were performed to identify and evaluate evidence for or against four propositions related to soyfoods and weight loss (Data from in vitro, animal, epidemiologic, and clinical studies were evaluated and summarized). (1) Certain soyfoods will improve weight and/or fat loss when fed at isolcaloric levels (similar calories given across experimental conditions, but not necessarily at a level to maintain current body weight); generally supportive evidence in animal studies, but there is no compelling support in human studies. (2) Certain soyfoods will improve weight and fat loss when included as part of a diet by affecting caloric intake; limited supportive evidence in animal and human studies. (3) Certain soyfoods will prevent/improve risk factors related to glucoregulatory function and cardiovascular health during weight loss; some evidence supporting this proposition, but additional evidence is needed before conclusions can be made. (4) Certain soyfoods will minimize the loss of bone mass during weight loss; no data available pertinent to this proposition. Limitations in existing data make it difficult to reach conclusions regarding these four propositions. Overall, the current data suggest that soyfoods are as good as other protein sources for promoting weight loss and there is a suggestive body of evidence that soyfoods may confer additional benefits, but results must be carefully interpreted and additional evidence is needed before making firm conclusions concerning soyfoods and weight loss. PMID- 18419672 TI - John Garrow (1929- ) treat obesity seriously! PMID- 18419676 TI - Prospects for the use of differentiation-modulating agents as adjuvant of photodynamic therapy for proliferative dermatoses. AB - Current interest in photodynamic therapy (PDT) in dermatology stems from its recognized success in dermatological oncology, straightforward approach, easy accessibility and low cost. PDT is a photochemistry-based modality in which a light-activated photosensitizer (PS) destroys tissue through oxygen-dependent and -independent mechanisms. Although PDT has been used in dermatology for several decades, its application has still not extended significantly into the routine management of neoplastic and proliferative dermatoses because of continuing issues with the selectivity of the PS for affected tissues. This review analyzes prospects for optimization of PDT for the management of dermatoses with defects in keratinocyte proliferation/differentiation, and discusses the use of differentiating agents that redirect metabolic utilization within cells and lead to high levels of PS accumulation. PMID- 18419677 TI - Clinical relevance of serum levels of matrix metallopeptidase-2, and tissue inhibitor of metalloproteinase-1 and -2 in patients with malignant melanoma. AB - The interaction and/or balance between matrix metallopeptidase (MMP)-2 and tissue inhibitor of metalloproteinase (TIMP)-2 in vivo may play important roles in the process of tumor growth, invasion and metastasis of malignant melanoma. In this study, we investigated the serum levels and immunohistochemical expression of MMP 2, TIMP-1 and TIMP-2 in patients with melanoma and analyzed the correlation with clinicopathological parameters. The level of serum MMP-2 in patients was significantly higher than that of the control. Moreover, the level of MMP-2 was significantly higher than that of the control in patients who were: (i) female; (ii) pT1 and pT4; (iii) with and without lymph node (LN) metastasis; (iv) in stage I and stage IV; (v) with and without recurrence; and (v) alive and dead. The level of serum TIMP-1 in patients with melanoma was significantly higher than that of the control. Among melanoma patients, the level of TIMP-1 with pT4 was significantly higher for patients who were: (i) pT1 and pT3; (ii) with LN metastasis (vs those without); (iii) in stage IV (vs those in stages I, II and III); and (iv) dead (vs those alive). The level of serum TIMP-2 in patients with melanoma was not different from the control. However, the level of TIMP-2 in patients with pT4 was significantly higher than for patients who were: (i) pT1, pT3 and control; (ii) with LN metastasis (vs those without metastasis and control); (iii) with stage IV (vs those in stages I and II and control); (iv) in recurrence (vs control); and (v) dead (vs those alive and control). These results suggest that increased serum levels of TIMP-1 and TIMP-2 reflected the extent of metastatic melanoma lesions, and that serum levels of TIMP-1 may be a new useful marker for melanoma progression. PMID- 18419678 TI - Expression of estrogen receptor beta in normal skin, melanocytic nevi and malignant melanomas. AB - Sex hormones are known to be associated with increases of melanocytes and melanin production in human skin. However, the expression of estrogen receptor (ERalpha) in melanocytic lesions has been controversial. In 1996, a new subset of estrogen receptor was cloned, and named estrogen receptor beta (ERbeta). We used immunohistochemical staining to characterize the expression of ERalpha and ERbeta in normal skin and in melanocytic lesions. Normal sebaceous glands and hair follicles were positive for ERalpha and ERbeta. Other adnexal structures and constituents in the skin were positive for ERbeta, but not for ERalpha. Melanocytic nevi and malignant melanomas were negative for ERalpha, but both were positive for ERbeta. The ubiquitous expression of ERbeta may play a fundamental role in various normal skin cells and melanocytic tumors. PMID- 18419679 TI - Case of anaphylaxis caused by ingestion of royal jelly. AB - Anaphylaxis is a severe form of allergic disease. Royal jelly is widely consumed in Japan, but a few cases of anaphylaxis caused by royal jelly have been reported. We encountered a 26-year-old Japanese woman who developed anaphylaxis after drinking a beverage of crude royal jelly including honey. She had a history of bronchial asthma, allergic rhinitis, allergic conjunctivitis, atopic dermatitis and food allergy (abalone). Prick tests were performed with the beverage of crude royal jelly including honey and with two other beverages including honey. Only the crude royal jelly beverage showed a positive reaction. An oral challenge test with the crude royal jelly beverage was not performed, but a similar test with a beverage including honey caused no symptoms. A positive response to the beverage of crude royal jelly was not observed in normal volunteers. A positive diagnosis of anaphylaxis due to royal jelly was made based on the positive prick test, systemic clinical symptoms and the negative prick tests in healthy volunteers. Moreover, the patient had no symptoms when taking lemon and orange, which were present as essences in the crude royal jelly beverage, and also had no response to honey after anaphylaxis. Increased consumption of royal jelly in health food supplements may increase the incidence of royal jelly-related allergic reactions. Therefore, royal jelly should be considered as a causative allergen in food-induced anaphylaxis. PMID- 18419680 TI - Case of adult T-cell leukemia with pulmonary involvement presenting as nodular shadows. AB - A 60-year-old man with a diagnosis of smoldering adult T-cell leukemia (ATL) had been treated successfully for 4 years with psoralen and ultraviolet A therapy, gamma-interferon, oral etoposide and sobuzoxane. He subsequently developed rapidly-growing skin nodules over his entire body. Chest X-ray and thoracic computed tomography showed nodular shadows in the right lower lung field and nodules in both lower lung lobes. Despite combined chemotherapy, he died. Upon autopsy, numerous nodules were found in the bilateral lower lobes; microscopically, the nodules were diffusely infiltrated by ATL cells. Our review of the published work found only two previously reported cases of ATL with pulmonary involvement manifested as nodular shadows. Herein, we present details on the third case. PMID- 18419681 TI - Case of hypereosinophilic syndrome with cutaneous necrotizing vasculitis. AB - A 25-year-old male presented with pruritic papules over his trunk and extremities. The pruritic eruption had appeared on the arms 5 months before and gradually spread over the trunk. Blood tests showed a white blood cell count of 13 760/microL with 42.2% eosinophils. A skin biopsy from the leg revealed necrotizing vasculitis in the upper dermis with prominent eosinophil infiltration in the dermis and subcutis. Liver dysfunction in accordance with eosinophilia was also seen. The underlying disorder causing the eosinophilia was not detected. We diagnosed this case as hypereosinophilic syndrome. Although systemic corticosteroid therapy using prednisolone was effective, the symptoms recurred during a reduction of prednisolone. The addition of cyclosporine resulted in improvement of his symptoms. Both drugs were successfully tapered without relapsing symptoms. Hypereosinophilic syndrome with cutaneous necrotizing vasculitis is a rare condition, and this case suggests that cyclosporine may be a useful therapeutic adjunct in facilitating steroid tapering. PMID- 18419682 TI - Chronic idiopathic macrocheilia associated with simple lip enlargement and salivary gland hyperplasia. AB - Chronic macrocheilia is a relatively rare condition of chronic lip enlargement. It can be cosmetically and functionally problematic, and often poses a diagnostic and therapeutic challenge to clinicians especially when its etiology is unclear. We describe a 40-year-old Korean man who presented with a 4-year history of lower lip swelling and was diagnosed as having chronic idiopathic macrocheilia after excluding other disease entities based on histopathological and clinical findings. We believe that the present case has a unique clinico-histopathological feature that differentiates it from other cases of chronic cheilitis, namely, simple lip swelling showing minor salivary gland hyperplasia only, suggesting the possibility of a new disease entity. PMID- 18419683 TI - Extravascular papillary endothelial hyperplasia of the palm masquerading as an angiosarcoma. AB - Cutaneous extravascular papillary endothelial hyperplasia (PEH) is a rare lesion presenting as a cutaneous mass and histologically mimicking angiosarcoma. Herein, we describe a patient who presented with a palm tumor that had developed after trauma. Histologically, the lesion was ulcerated and showed proliferation of atypical epithelioid cells with nuclear enlargement and mitosis. Vascular proliferation was evident, and these were positive for CD31 and CD34, with approximately 20% positivity for MIB-1. We diagnosed this case as an extravascular papillary endothelial hyperplasia because there was no evidence of invasion into the surrounding tissue and elastica staining proved it did not have a vascular structure. To our knowledge, cutaneous extravascular PEH has not yet been reported in the published work. PMID- 18419684 TI - Clavus detected incidentally by positron emission tomography with computed tomography. AB - This is the first report showing that positron emission tomography with computed tomography (PET/CT) can detect clavus. A 37-year-old woman diagnosed with epithelioid sarcoma on her left inguinal region was investigated by whole-body PET/CT. The imaging showed some 18F-fluorodeoxyglucose uptake at the right lateral foot. There had been a clavus in exactly the same region. Every dermatologist, radiologist and oncologist needs to know about this finding for making an accurate diagnosis. PMID- 18419685 TI - Granulomatous cheilitis successfully treated with roxithromycin. PMID- 18419686 TI - Case of premenstrual syndrome inducing monthly episodes of vesiculobullous eruptions on the face. PMID- 18419687 TI - Adult-onset Still's disease complicated by multiple cranial nerve palsy and hypothyroidism. PMID- 18419688 TI - Adverse skin reactions associated with hepatitis C treatment: comment. PMID- 18419690 TI - Fatal pulmonary renal syndrome and treatment in adults with Schonlein-Henoch purpura: comment. PMID- 18419692 TI - Abstracts of the The Royal Australasian College of Surgeons Annual Scientific Congress. May 12-16, 2008. Hong Kong. PMID- 18419693 TI - Regulating medical bodies? The consequences of the 'modernisation' of the NHS and the disembodiment of clinical knowledge. AB - The aim of this paper is to explore the consequences of modernisation and regulatory processes for the everyday lives of doctors working the UK National Health Service. We do this by reporting on interview data generated as part of a qualitative investigation into the working lives of 47 doctors. The analysis of the empirical findings is informed by two literatures: that which has sought to theorise the contemporary thrust of regulation and audit and that which has developed a sociology of embodiment. Doctors' views are presented in relation to four areas of work which hav--in the loosest sense of the word--been subject to regulation. Drawing on work from the sociology of embodiment we argue that changes in the institutional and cultural context of medical work could be altering both the 'field' and the 'habitus'--to use Bourdieu's terms--of medicine, with a consequence that medical knowledge is becoming less embodied. PMID- 18419694 TI - A genetic future for coronary heart disease? AB - This paper is concerned with changing conceptions of genetic disease. It is based on an analysis of biomedical literature and focuses on the treatment of coronary heart disease (CHD) in four published commentary papers. The aim of this analysis is to explore the ways in which CHD is constructed as genetic and the place of genetic discourses in the wider set of ideas that circulate about the disease. This analysis is then used to consider some of the claims of the geneticisation thesis (Lippman 1991, 1992). The analysis suggests that a genetic vision for understanding and managing CHD has emerged, which has many of the hallmarks of the geneticisation imagined by Lippman. However, a number of alternative and competing models of CHD are also supported within the biomedical discourse. These are related to the different disciplines with a stake in the field of CHD, and their struggles for authority. In conclusion, it is suggested that the geneticisation thesis, as a universal claim, is at odds with the diffuse and distributed nature of biomedical knowledge and practice. Rather than analysing geneticisation in a literal way, it may be more fruitful to see the thesis, itself, as a form of boundary work (Gieryn 1983). PMID- 18419695 TI - Like a prayer: the role of spirituality and religion for people living with HIV in the UK. AB - Over 40,000 people are now living with diagnosed HIV in the UK. There is, however, uncertainty about how people with HIV use religion or spirituality to cope with their infection. Adopting a modified grounded theory approach, we analysed individual and group interviews with the people most affected by HIV in the UK: black African heterosexual men and women and gay men (mostly white). For the majority of black African heterosexual men and women in our study, religion was extremely important. We found that gay men in the study were less religious than black Africans, although many were spiritual in some way. Black African individuals constructed their spiritual narratives as largely Christian or collective, while gay men described more individualistic or 'New Age' approaches. We developed a six-level heuristic device to examine the ways in which prayer and meditation were deployed in narratives to modulate subjective wellbeing. These were: (i) creating a dialogue with an absent counsellor; (ii) constructing a compassionate 'life scheme'; (iii) interrupting rumination; (iv) establishing mindfulness; (v) promoting positive thinking, and (vi) getting results. That people with HIV report specific subjective benefits from prayer or meditation presents a challenge to secular healthcare professionals and sociologists. PMID- 18419696 TI - BJU International. Editor's comment. PMID- 18419697 TI - Is the biopsy Gleason score important in predicting outcomes for patients after radical prostatectomy once the pathological Gleason score is known? AB - OBJECTIVE: To evaluate whether specific preoperative variables might better predict the concordance between biopsy and radical prostatectomy (RP) Gleason grade, and to assess the effect of the biopsy Gleason score (bGS) when controlling for the pathological GS (pGS) on clinical outcomes in patients undergoing RP. PATIENTS AND METHODS: Between 1989 and 1998, 1088 men had RP at our institution, with a median follow-up of 56 months. To evaluate the independent effect of bGS within categories of pGS, we stratified the sample by pGS (three categories; 15), EPO measurement was obtained on admission, during the first 3 days and then when Hb level was measured. Maximal EPO level (EPOmax) and minimal Hb level (Hbmin) during the ICU stay was determined for all patients. RESULTS: One hundred and seventy-one consecutives patients were included (440 EPO measurements). Seventy-nine patients (46.2%) showed an increased value (> or =25 UI/l) EPOmax value. Most EPOmax values were observed early after the trauma [within 4 days for 63 patients (82.8%)]. Plotting EPOmax to Hbmin values show that a threshold Hbmin value of 105 g/l best discriminated patients with and without an elevated EPO secretion. Less than 10% of the patients with Hbmin<105 g/l did not increase their EPO secretion. CONCLUSION: In severely traumatized patients a marked response to acute anaemia is observed in most patients. In our study, Hb threshold for a significant EPO secretion following post-traumatic acute anaemia was 105 g/l. The peak level was achieved early in the course of the anaemia. PMID- 18419713 TI - Impaired erythropoiesis after haemorrhagic shock in mice is associated with erythroid progenitor apoptosis in vivo. AB - OBJECTIVE: Multiply traumatised patients often suffer from blood loss and from subsequent therapy-resistant anaemia, possibly mediated by apoptosis, necrosis, or humoral factors. Therefore, the underlying mechanisms were investigated in bone marrow (BM) and peripheral blood in a murine resuscitated haemorrhagic shock (HS) model. METHODS: In healthy male mice, pressure-controlled HS was induced for 60 min. The BM was analysed for Annexin-V, 7-amino-actinomycin D, apoptotic enzymes (caspases-3/7, -8, and -9), expression of death receptors (CD120a, CD95), mitochondrial proteins (Bax, Bcl-2, Bcl-x), as well as erythropoietin (EPO) receptor (EPO-R). Blood cell count, peripheral EPO, and tumour necrosis factor alpha response were additionally monitored. RESULTS: Twenty-four and 72 h after HS, EPO and EPO-R were strongly up-regulated in peripheral blood and BM, respectively. Decreasing numbers of erythroid progenitors in BM after HS correlated with significant apoptotic changes confirmed by increased caspases 3/7, -8, -9 activity in total BM, death receptor CD95 and CD120a expression on erythroid progenitors, and down-regulated mitochondrial Bcl-2 expression in total BM. Erythroid progenitors in peripheral blood were found to be increased after 72 h. CONCLUSION: Despite the massive EPO response and up-regulation of EPO-R, BM erythroblasts (EBs) decreased. This could be due to deficient maturation of erythroid progenitors. Furthermore, the increased intrinsic and extrinsic apoptosis activation suggests programmed death of erythroid progenitors. We propose that both apoptosis and negatively regulated erythropoiesis contribute to BM dysfunction, while erythroid progenitor egress plays an additional role. PMID- 18419714 TI - Renal cytokine profile in an endotoxemic porcine model. AB - INTRODUCTION: In animals exposed to acute endotoxemia with lipopolysaccharide (LPS), high levels of cytokines are found in the kidney. The objective of this study is to determine whether the high renal content of TNF-alpha, IL-1beta, IL 10 and IL-1 receptor antagonist (IL-1ra) is due to glomerular filtration and reabsorption, or whether the cytokines are produced locally in the kidney. METHODS: Eighteen anesthetized and mechanically ventilated pigs (35-43 kg) were randomized into two groups: Group 1 (n=12) LPS infusion for 360 min and Group 2 (n=6) control pigs, no treatment. At 360 min, the pigs were euthanized and tissue samples from the kidneys were obtained. Localization of the cytokines was determined by immunohistochemistry and double immunofluorescence (dIF). RESULTS: Pigs exposed to endotoxemia showed increased accumulation of leukocytes and increased protein expression of TNF-alpha and IL-1beta when compared with controls. dIF showed that TNF-alpha-positive cells co-localized with both endothelial and mesangial cells in the glomeruli. Furthermore, the endothelial cells of the cortical arterioles were positive for IL-1beta. TNF-alpha and IL 1beta staining were absent in renal tubular cells. A positive signal for IL-10 was detected at the tubular brush border while IL-1ra was detected in the glomerulus and in the tubular cells. CONCLUSION: LPS-induced endotoxemia increased TNF-alpha and IL-1beta protein expression and leukocyte accumulation in the kidneys. The results indicate that the increased levels of the pro inflammatory cytokines TNF-alpha and IL-1beta are caused by a local production in the kidneys while the anti-inflammatory cytokines IL-10 and IL-1ra are filtrated and reabsorbed in the tubuli. PMID- 18419716 TI - Insulin alters cytokine content in two pivotal organs after brain death: a porcine model. AB - BACKGROUND: To optimize the quantity and quality of organs available for transplantation, it is crucial to gain further insight into the treatment of brain dead organ donors. In the current study we hypothesized that insulin treatment after brain death alters cytokine content in the heart, liver, and kidney. METHODS: Sixteen brain dead pigs (35-40 kg) were treated with either (1) no insulin [brain dead without insulin treatment treatment (BD)], or (2) insulin infusion intravenously (i.v.) at a constant rate of 0.6 mU/kg/min during 360 min [brain dead with insulin treatment (BD+I)]. Blood glucose was clamped at 4.5 mmol/l by infusion of 20% glucose. Blood samples for insulin, glucose, catecholamines, free fatty acids, and glucagon were obtained during the experimental period. Six hours after brain death biopsies were taken from the heart, liver, and kidney. These were analyzed for cytokine mRNA and proteins [tumor necrosis factor-alpha (TNF-alpha), interleukin (IL)-6, and IL-10]. RESULTS: The BD+I compared with the BD animals had lower IL-6 concentrations in the right ventricle of the heart (P=0.001), in the renal cortex (P=0.04) and in the renal medulla (P=0.05), and lower IL-6 mRNA in the renal medulla (P=0.0002). Furthermore, the BD+I animals had lower concentrations in the renal medulla of IL 10 (P=0.01), and tended to have lower TNF-alpha in the renal cortex (P=0.06) than the BD animals. In the right ventricle of the heart TNF-alpha mRNA and IL-10 mRNA were higher in the BD+I than in the BD group (P=0.002 and 0.004). CONCLUSION: Insulin has anti-inflammatory effects on cytokine concentration in the heart and kidney after brain death. PMID- 18419715 TI - Does brain death induce a pro-inflammatory response at the organ level in a porcine model? AB - BACKGROUND: Organs from brain-dead donors have a poorer prognosis after transplantation than organs from living donors. A possible explanation for this is that brain death might initiate a systemic inflammatory response, elicited by a metabolic stress response or brain ischemia. The aim of this study was to investigate the effect of brain death on the cytokine content in the heart, liver, and kidney. In addition, the metabolic and hemodynamic response caused by brain death was carefully registered. METHODS: Fourteen pigs (35-40 kg) were randomized into two groups (1) eight brain-dead pigs and (2) six pigs only sham operated. Brain death was induced by inflation of an epidurally placed balloon. Blood samples for insulin, glucose, catecholamine, free fatty acids (FAA), and glucagon were obtained during the experimental period of 360 min. At the conclusion of the experiment, biopsies were taken from the heart, liver, and kidney and were analyzed for cytokine mRNA and proteins [tumor necrosis factor alpha (TNF-alpha), interleukin (IL)-6, and IL-10). RESULTS: We found a dramatic response to brain death on plasma levels of epinephrine (P=0.004), norepinephrine (P=0.02), FAA (P=0.0001), and glucagon (P=0.0003) compared with the sham group. There was no difference in cytokine content in any organ between the groups. CONCLUSION: In this porcine model, brain death induced a severe metabolic response in peripheral blood. At the organ level, however, there was no difference in the cytokine response between the groups. PMID- 18419718 TI - Maximal hysteresis: a new method to set positive end-expiratory pressure in acute lung injury? AB - BACKGROUND: No methods are superior when setting positive end-expiratory pressure (PEEP) in acute lung injury (ALI). In ALI, the vertical distance (hysteresis) between the inspiratory and expiratory limbs of a static pressure-volume (PV) loop mainly indicates lung recruitment. We hypothesized that PEEP set at the pressure where hysteresis is 90% of its maximum (90%MH) would give similar oxygenation, but less cardiovascular depression than PEEP set at the pressure at lower inflection point (LIP) on the inspiratory limb or at the point of maximal curvature (PMC) on the expiratory limb in ALI. METHODS: In 12 mechanically ventilated pigs, ALI was induced in a randomized fashion by lung lavage, lung lavage plus injurious ventilation, or by oleic acid. From a static PV loop obtained by an interrupted low-flow method, the pressures at LIP [25 (25, 25) cmH(2)O, mean and 25, 75 percentiles], at PMC [24 (20, 24) cmH(2)O], and at 90% MH [19 (18, 19) cmH(2)O] were determined and used for the PEEP-settings. We measured lung inflation (by computed tomography), end-expiratory lung volume (EELV), airway pressures, compliance of the respiratory system (Crs), blood gases, cardiac output and arterial blood pressure. RESULTS: There were no differences between the PEEP settings in EELV or oxygenation, but the 90%MH setting gave lower end-inspiratory pause pressure (P<0.025), higher Crs (P<0.025), less hyper-aeration (P<0.025) and better maintained hemodynamics. CONCLUSION: In this porcine lung injury model, PEEP set at 90% MH gave better lung mechanics and hemodynamics, than PEEP set at PMC or LIP. PMID- 18419717 TI - Sirtinol attenuates hepatic injury and pro-inflammatory cytokine production following trauma-hemorrhage in male Sprague-Dawley rats. AB - BACKGROUND: Although studies have demonstrated that sirtinol administration following adverse circulatory conditions is known to be protective, the mechanism by which sirtinol produces the salutary effects remains unknown. We hypothesized that sirtinol administration in male rats following trauma-hemorrhage decreases cytokine production and protects against hepatic injury. METHODS: Male Sprague Dawley rats underwent trauma-hemorrhage (mean blood pressure 40 mmHg for 90 min, then resuscitation). A single dose of sirtinol (1 mg/kg of body weight) or vehicle was administered intravenously during resuscitation. Twenty-four hours thereafter, tissue myeloperoxidase (MPO) 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 liver and plasma alanine aminotransferase (ALT) concentrations were measured (n=6 Sprague-Dawley rats/group). RESULTS: Trauma-hemorrhage increased hepatic MPO activity, CINC-1, CINC-3, ICAM-1, and IL-6 levels and plasma ALT concentrations. These parameters were significantly improved in the sirtinol-treated rats subjected to trauma-hemorrhage. CONCLUSION: The salutary effects of sirtinol administration on attenuation of hepatic injury following trauma-hemorrhage are, at least in part, related to reduction of pro-inflammatory mediators. PMID- 18419720 TI - Effects of halothane, isoflurane, and sevoflurane on lipid peroxidation following experimental closed head trauma in rats. AB - BACKGROUND: In a rat closed head trauma model we examined both the time course of lipid peroxidation and the effects of halothane, isoflurane, and sevoflurane on it by analysis of malondialdehyde (MDA) formation. METHODS: Animals were divided randomly into five groups: sham-operated (SO), n=18; control-closed head trauma to left frontal pole, n=18; closed head trauma model+halothane, n=18; closed head trauma model+isoflurane, n=18; and closed head trauma model+sevoflurane, n=18. Halothane, isoflurane, or sevoflurane were applied 15 min after trauma for 30 min. Rats were euthanized 1,3, and 5 h after the inhalation agents. Brain tissue samples were taken 5 mm from the left and right frontal poles. MDA was considered to reflect the degree of lipid peroxidation. RESULTS: MDA concentrations were greater in the control, halothane, sevoflurane, and isoflurane groups than in SO animals (P<0.001). No statistical difference between the hemispheres was found between the halothane, isoflurane, or sevoflurane groups, but MDA levels were lower with isoflurane than in the halothane, sevoflurane, and control groups at 1, 3, and 5 h (P<0.001). MDA levels were higher as compared with the halothane and sevoflurane groups at 1 h but not at 3 or 5 h (P<0.001). CONCLUSION: MDA levels with the isoflurane group were lower than in the other trauma groups, which suggest that isoflurane, given after closed head trauma, might be protective against lipid peroxidation of cerebral injury. PMID- 18419719 TI - Isoflurane attenuates myoglobin release during ischemic and/or reperfusion periods. AB - BACKGROUND: Recently, we have developed cardiac microdialysis for detection of protein leakage from the injured myocardium. We examined whether the exposures to isoflurane would exert a beneficial effect on myocardial injury caused by ischemia or reperfusion. METHODS: A dialysis probe was implanted into the left ventricle free wall in the rabbits. The dialysate myoglobin level served as an index of myocardial interstitial myoglobin levels. Rabbits were randomly assigned to one of three groups: (1) without exposure to isoflurane (vehicle, n=6), (2) inhale 1 MAC isoflurane once for 30 min (ISO30-1, n=6), and (3) twice for 30 min (ISO30-2, n=6). All rabbits underwent 30 min of coronary occlusion and 60 min of reperfusion. To determine whether the isoflurane induced myocardial protection against chemical hypoxia, sodium cyanide (30 mM) was administered and dialysate myoglobin levels were measured with (n=6) and without pre-exposure to isoflurane twice for 30 min (n=6). RESULTS: In all three groups dialysate myoglobin levels were increased by coronary occlusion and furthermore augmented by reperfusion. In comparison with the vehicle group, the ISO30-1 group suppressed only the increase in the dialysate myoglobin level during reperfusion. The ISO30-2 group suppressed during both the ischemic and reperfusion periods. Cyanide induced increases in dialysate myoglobin levels. These increments in dialysate myoglobin levels were suppressed by repeated exposure to isoflurane. CONCLUSION: Repeated exposure to isoflurane suppressed myocardial myoglobin release caused by both ischemia and reperfusion injury. Isoflurane may provide protection against myocardial ischemia/reperfusion and hypoxic injuries. PMID- 18419722 TI - Ketamine blocks enhancement of spinal long-term potentiation in chronic opioid treated rats. AB - BACKGROUND: Long-term opioid treatment is associated with the development of hyperalgesia. In a rat model we wanted to study if chronic opioid treatment changed the induction and maintenance of spinal long-term potentiation (LTP), a form of hyperexcitability in the spinal cord. We also wanted to investigate if the clinically available NMDA receptor antagonist ketamine inhibited the effect of chronic opioid treatment on LTP. METHODS: The animals were randomized into four groups (saline, morphine 20 mg/kg/day, ketamine 20 mg/kg/day, morphine 20 mg/kg/day and ketamine 20 mg/kg/day). Drugs were given as continuous subcutaneous infusions by means of osmotic minipumps. After 7 days of treatment and during ongoing treatment single unit extracellular recordings were made from the lumbar deep dorsal horn under urethane anesthesia. Single electrical stimuli were applied to the sciatic nerve, and the C-fiber evoked responses of WDR neurons were recorded before and during 3 h following low frequency (3 Hz) electrical conditioning stimulation. RESULTS: The potentiation of C-fiber evoked responses by conditioning stimulation was significantly increased in the morphine-treated group compared to the saline group, while there was no significant difference between the saline, the ketamine and the morphine/ketamine groups. The potentiated responses in the morphine/ketamine group were significantly reduced compared to the morphine group (P=0.01). CONCLUSION: Our results indicate that animals treated with long-term opioid show amplification of stimulus-induced central sensitisation compared to opioid naive animals. Ketamine inhibited the morphine-induced enhancement of LTP, supporting the role of ketamine in prevention of opioid induced hyperalgesia. PMID- 18419721 TI - Effect of paracetamol and coxib with or without dexamethasone after laparoscopic cholecystectomy. AB - BACKGROUND: Pain after laparoscopic cholecystectomy (LCC) is multifactorial. Effective post-operative pain control is necessary in LCC performed as day-case surgery. We studied the efficacy of paracetamol or valdecoxib with or without dexamethasone after LCC. METHODS: One hundred sixty patients were randomized to four groups of 40 patients. Groups 1 and 3 received parecoxib 40 mg intravenously (IV) during surgery and valdecoxib 40 mg x 1 per os (PO) for 7 post-operative days. Groups 2 and 4 received paracetamol 1 g x 4 IV during surgery and 1 g x 4 PO for 7 days. In addition, Groups 3 and 4 were given dexamethasone 10 mg IV intra-operatively. Propofol and remifentanil were used during surgery. The patients were given oxycodone 0.05 mg/kg IV in phase 1 post-anaesthesia care unit (PACU 1) or 0.15 mg/kg PO in phase 2 post-anaesthesia care unit (PACU 2) as needed to keep visual analogue scale <3/10. The patients were supplied with the study drugs for 7 post-operative days. RESULTS: Pain intensity, nausea and the need of oxycodone in phase 1 PACU were similar in all groups. Dexamethasone reduced the need of oral oxycodone in phase 2 PACU (7.0 +/- 1.0 mg vs. 9.1 +/- 1.0 mg, P<0.05). Pain intensity was similar in all groups at home. More patients in the parecoxib/valdecoxib groups needed rescue medication on the 1st post operative day (P<0.001) than paracetamol-treated patients. CONCLUSION: Paracetamol was as effective as parecoxib/valdecoxib for pain after LCC. Dexamethasone decreased the need of oxycodone in phase 2 PACU. The effect of dexamethasone was similar in paracetamol and parecoxib/valdecoxib patients. PMID- 18419723 TI - Increased plasma glutamate, glycine, and arginine levels in complex regional pain syndrome type 1. AB - BACKGROUND: Various inflammatory mediators have been identified as potential contributors to complex regional pain syndrome type 1 (CRPS1), but these mediators do not entirely explain certain manifestations of the syndrome, such as pain. The objective of this study was to investigate the role of amino acids in the pathogenesis of CRPS1. METHODS: We used HPLC to determine plasma concentrations of 16 amino acids, especially those related to the NMDA receptor (e.g., glutamate and glycine) and nitric oxide (NO) synthesis (e.g., arginine and citrulline) in patients with CRPS1 (n=64) and age- and sex-matched healthy controls (n=51). Patients rated pain intensity (visual analog scale) and the subjective experience of pain intensity (McGill Pain Questionnaire). Psychological dysfunction was assessed using the SCL-90. RESULTS: Relative to controls, in CRPS1 patients, plasma levels of glutamate, arginine, taurine, and glycine were increased, and plasma levels of glutamine and the ratio of citrulline to arginine were decreased. Remarkably, in CRPS1 patients there was a highly significant inverse correlation between glutamine and glutamate, although the sum of molar concentrations of glutamate and glutamine remains unchanged. Subjective measures of pain and indicators of psychoneuroticism and emotional instability did not correlate with amino acid levels. CONCLUSION: This study shows for the first time a pronounced increase in amino acid levels in this chronic pain syndrome. The marked differences in glutamate, glutamine, glycine, taurine and arginine levels between patients and controls suggest the involvement of both the NDMA receptor and the endothelium-dependent arginine-NO system in CRPS1. PMID- 18419724 TI - Spinal 2-chloroprocaine: effective dose for ambulatory surgery. AB - BACKGROUND: There is an interest in finding a safe, short-acting spinal anaesthetic, suitable for ambulatory surgery. In this prospective study, we evaluated the effective dose of plain 2-chloroprocaine (2-CP) for lower limb surgery, including knee arthroscopy and saphenectomy. METHODS: Sixty-four ASA physical status I-III patients undergoing elective lower limb surgery were randomly allocated to one of the four local anaesthetic groups for spinal anaesthesia in a double-blind manner. The patients (n=16 patients in each group) received 35, 40, 45 or 50 mg of 10 mg/ml isobaric 2-CP. RESULTS: In all patients, anaesthesia was sufficient for the planned surgery. The median peak block height (T9) was similar in all four groups (P=0.66). Time to complete sensory block regression was faster in the 35 mg group (111 min, mean) and in the 40 mg group (108 min) than in the 50 mg group (134 min, P=0.005). No differences in time to complete motor block regression were observed (P=0.3). Home discharge time was faster in the 35 mg group (123 min) and in the 40 mg group (122 min) than in the 50 mg group (165 min, P=0.001). No complications related to spinal anaesthesia were observed and no transient neurologic symptoms (TNS) were reported at the 3 day follow-up. CONCLUSION: Spinal 2-CP, 10 mg/ml 35, 40, 45 and 50 mg provide reliable sensory and motor block for ambulatory surgery, while reducing the dose of 2-CP to 35 and 40 mg resulted in a spinal block of faster ambulation. PMID- 18419725 TI - Laparoscopic colon surgery: unreliability of end-tidal CO2 monitoring. AB - BACKGROUND: The relatively good haemodynamic and respiratory tolerance to abdominal CO(2) insufflation has mostly been observed in healthy patients during short-lasting laparoscopic procedures. End-tidal CO(2) pressure (PetCO(2)) has been shown to be a reliable method to assess arterial CO(2) (PaCO(2)) in the absence of cardio-respiratory disease in this setting. However, no study has investigated whether PetCO(2) is accurately related to PaCO(2) during laparoscopic colon surgery. Indeed, these procedures last longer, prolonging the pneumoperitoneum and requiring a Trendelenburg position. The aim of the present study was to measure the PaCO(2)-PetCO(2) difference over time in patients undergoing laparoscopic colon surgery and to determine whether PaCO(2) is reliably assessed by PetCO(2). METHODS: Forty consecutive patients (ASA I and II) scheduled for laparoscopic colon surgery were anaesthetized and ventilated to obtain a PetCO(2) between 4.0 and 5.5 kPa. After initiation of CO(2) insufflation, PaCO(2) and PetCO(2) were recorded every 30 min during surgery. RESULTS: No complication was observed during anaesthesia. The mean arterial pressure increased significantly after CO(2) insufflation and remained steady up to the end of pneumoperitoneum. The heart rate remained stable over time. The relation between PaCO(2) and PetCO(2) was not constant among patients and increased over time within the same patients. The R(2) values fluctuated and did not show a constant correlation between PaCO(2) and PetCO(2). CONCLUSION: The correlation between PaCO(2) and PetCO(2) during laparoscopic colon surgery is inconsistent mainly due to inter- and intra-individual variability. PMID- 18419726 TI - Effects of fentanyl on gastric myoelectrical activity: a possible association with polymorphisms of the mu-opioid receptor gene? AB - BACKGROUND: Opioids have inhibitory effects on gastric motility, but the mechanism is far from clear. Electrical slow waves in the stomach determine the frequency and the peristaltic nature of gastric contractions. The primary aim of this study was to investigate the effects of the opioid fentanyl on gastric myoelectric activity. As there were large variations between the subjects, we investigated whether the variation was correlated to single nucleotide polymorphisms (SNP) of the mu-opioid receptor (MOR) gene. METHODS: We used cutaneous multichannel electrogastrography (EGG) to study myoelectrical activity in 20 patients scheduled for elective surgery. Fasting EGG was recorded for 30 min, followed by intravenous administration of fentanyl 1 microg/kg and subsequent EGG recording for 30 min. Spectral analysis of the two recording periods was performed and the variables assessed were dominant frequency (DF) of the EGG and its power (DP). Genetic analysis of the SNP A118G and G691C of the MOR gene was performed with the polymerase chain reaction technique. RESULTS: There was a significant reduction in DF and DP after intravenous fentanyl. However, there was a large variation between the patients. In eight subjects EGG was unaffected, five subjects had a slower DF (bradygastria) and in six subjects the slow waves disappeared. We found no correlation between the EGG outcome and the presence of A118G or G691C in the MOR gene. CONCLUSIONS: Fentanyl inhibited gastric myoelectrical activity in about half of the subjects. The variation could not be explained by SNP in the MOR gene. Because of small sample size, the results must be regarded as preliminary observations. PMID- 18419727 TI - Carbon monoxide in biological systems: an experimental and clinical study. PMID- 18419728 TI - Acute beta-adrenergic blockade: an unusual cause of suppression of spectral entropy values during isoflurane anaesthesia. PMID- 18419729 TI - Tongue flap: a challenge to the anesthesiologist. PMID- 18419730 TI - Failure to deflate an endotracheal cuff. PMID- 18419732 TI - Ultra-sound guided sciatic nerve block combined with lumbar plexus block for infra-inguinal artery bypass graft surgery. PMID- 18419731 TI - Cardiopulmonary bypass as a risk factor in paediatric cardiac surgical patients. PMID- 18419733 TI - Use of saline infusion instead of gel for ultrasound-guided (USG) central venous cannulation. PMID- 18419734 TI - Overexpression of Arabidopsis thaliana tryptophan synthase beta 1 (AtTSB1) in Arabidopsis and tomato confers tolerance to cadmium stress. AB - Tryptophan (Trp) is an essential amino acid in humans, and in plants, it plays a major role in the regulation of plant development and defence responses. However, little is known about Trp-mediated cadmium (Cd) tolerance. Gene expression analysis showed that Arabidopsis thaliana tryptophan synthase beta 1 (AtTSB1) is up-regulated in plants treated with Cd; hence, we investigated whether this gene is involved in Cd tolerance. Exogenous application of Trp to wild-type Arabidopsis enhances Cd tolerance. Cd tolerance in the Trp-overproducing mutant trp5-1 was associated with high chlorophyll levels and low lipid peroxidation, as indicated by malondialdehyde 4-hydroxyalkenal level, whereas the wild-type developed symptoms of severe chlorosis. Moreover, the Trp-auxotroph mutant trp2-1 was sensitive to Cd. CaMV 35S promoter-driven AtTSB1 enhanced Trp accumulation and improved Cd tolerance in transgenic Arabidopsis and tomato plants without increasing the level of Cd. Moreover, reverse transcription-polymerase chain reaction confirmed that enhanced level of Trp in AtTSB1 transgenic Arabidopsis plants affected the expression of AtZIP4 and AtZIP9 metal transporters, which interfered with Cd ion trafficking, a mechanism of transcriptional regulation that does not exist in wild-type plants. Overexpression of AtTSB1 in transgenic tomato also produced higher Trp synthase-beta enzyme activity than that in wild type plants. These results implicate that Trp could be involved in Cd defence. PMID- 18419735 TI - Phosphatidylinositol 4-phosphate accumulates extracellularly upon xylanase treatment in tomato cell suspensions. AB - Various phosphoinositides have been implicated in plant defence signalling. Until now, such molecules have been exclusively related to intracellular signalling. Here, evidence is provided for the detection of extracellular phosphatidylinositol 4-phosphate (PI4P) in tomato cell suspensions. We have analysed and compared the intracellular and extracellular phospholipid profiles of [(32)P(i)]-prelabelled tomato cells, challenged with the fungal elicitor xylanase. These phospholipid patterns were found to be different, being phosphatidylinositol phosphate (PIP) the most abundant phospholipid in the extracellular medium. Moreover, while cells responded with a typical increase in phosphatidic acid and a decrease in intracellular PIP upon xylanase treatment, extracellular PIP level increased in a time- and dose-dependent manner. Using two experimental approaches, the extracellular PIP isoform was identified as PI4P. Addition of PI4P to tomato cell suspensions triggered the same defence responses as those induced by xylanase treatment. These include production of reactive oxygen species, accumulation of defence-related gene transcripts and induction of cell death. We demonstrate that extracellular PI4P is accumulated in xylanase elicited cells and that exogenous application of PI4P mimics xylanase effects, suggesting its putative role as an intercellular signalling molecule. PMID- 18419736 TI - NaCl-induced changes in cytosolic free Ca2+ in Arabidopsis thaliana are heterogeneous and modified by external ionic composition. AB - Increases in cytosolic free Ca(2+) ([Ca(2+)](cyt)) are common to many stress activated signalling pathways, including the response to saline environments. We have investigated the nature of NaCl-induced [Ca(2+)](cyt) signals in whole Arabidopsis thaliana seedlings using aequorin. We found that NaCl-induced increases in [Ca(2+)](cyt) are heterogeneous and mainly restricted to the root. Both the concentration of NaCl and the composition of the solution bathing the root have profound effects on the magnitude and dynamics of NaCl-induced increases in [Ca(2+)](cyt). Alteration of external K(+) concentration caused changes in the temporal and spatial pattern of [Ca(2+)](cyt) increase, providing evidence for Na(+)-induced Ca(2+) influx across the plasma membrane. The effects of various pharmacological agents on NaCl-induced increases in [Ca(2+)](cyt) indicate that NaCl may induce influx of Ca(2+) through both plasma membrane and intracellular Ca(2+)-permeable channels. Analysis of spatiotemporal [Ca(2+)](cyt) dynamics using photon-counting imaging revealed additional levels of complexity in the [Ca(2+)](cyt) signal that may reflect the oscillatory nature of NaCl induced changes in single cells. PMID- 18419737 TI - Transcript profiling indicates that the absence of PsbO affects the coordination of C and N metabolism in Synechocystis sp. PCC 6803. AB - Transcript profiling of nitrate-grown Synechocystis sp. PCC 6803 PsbO-free mutant cells in comparison to wild-type (WT) detected substantial deviations. Because we had previously observed phenotypical differences between Synechocystis sp. PCC 6803 WT and its corresponding PsbO-free mutant when cultivated with l-arginine as sole N source and a light intensity of 200 mumol photons m(-2) s(-1), we also performed transcript profiling for both strains grown either with nitrate or with l-arginine as sole N source. We observed a total number of 520 differentially regulated transcripts in Synechocystis WT because of a shift from nitrate- to l arginine-containing BG11 medium, while we detected only 13 differentially regulated transcripts for the PsbO-free mutant. Thus, the PsbO-free Synechocystis mutant had already undergone a preconditioning process for growth with l-arginine in comparison to WT. While Synechocystis WT suffered from growth with l-arginine at a light intensity of 200 mumol photons m(-2) s(-1), the PsbO-free mutant developed only a minor stress phenotype. In summary, our results suggest that the absence of PsbO in Synechocystis affects the coordination of photosynthesis/respiration and l-arginine metabolism through complex probably redox-mediated regulatory pathways. In addition, we show that a comparison of the transcriptomes of nitrate-grown Synechococcus elongatus PCC 7942 WT cells and its corresponding PsbO-free mutant cells resulted in only a few differentially regulated transcripts between both strains. The absence of the manganese/calcium stabilizing PsbO protein of PSII with an assigned regulatory function for photosynthetic water oxidation causes bigger changes in the transcriptome of the permissive photoheterotrophically growing Synechocystis sp. PCC 6803 than in the transcriptome of the obligate photoautotrophically growing S. elongatus PCC 7942. PMID- 18419738 TI - Photosynthetic redox regulation of the plastocyanin promoter in tobacco. AB - Persistent light quality gradients in dense plant populations induce imbalances in the distribution of excitation energy between the photosystems. Plants counteract such conditions by re-adjusting the stoichiometry of photosystems, which involves control of photosynthesis gene expression both in chloroplasts and in the nucleus. Decisive control parameters are redox signals from the photosynthetic electron transport chain, one prominent is the plastoquinone (PQ) pool. In a recent study, a plastocyanin (PC)-promoter::beta-glucuronidase reporter gene construct in tobacco demonstrated reversible redox regulation in response to varying light qualities. Here, northern and Western analyses demonstrate that this promoter regulation also accounts for the accumulation of the endogenous tobacco PetE gene transcripts and the protein amounts of the encoded PC. Hence, the reporter gene construct reflects the natural regulation of this nuclear gene in tobacco. In kinetic experiments, the response of the construct to either oxidation or reduction of the PQ pool was tested by defined light quality shifts. The construct displayed upregulation in response to a reduction signal and downregulation in response to an oxidation signal, both with a half-time of about 24 h. The response was finished after 48 h. DCMU application abolished the upregulation in response to the reduction signal, indicating the dependence on thylakoid membrane electron transport. To study the redox responsive promoter region in more detail, several promoter deletion constructs were tested for their responsiveness. All constructs displayed a reversible response to light-induced oxidation and reduction signals; however, a minimal promoter region localised between -168 to -79 bp upstream of the transcription start site was sufficient to confer this redox regulation. This indicates that photosynthetic redox signals act on distinct regions in the PC promoter in a manner independent from photoreceptors and upstream cis elements conferring high basic expression in the light. PMID- 18419739 TI - Inhibition of ammonium assimilation restores elongation of seminal rice roots repressed by high levels of exogenous ammonium. AB - Elongation of seminal and lateral roots of rice seedlings was markedly inhibited by high ammonium levels in growth medium. However, high exogenous nitrate concentrations had little inhibitory effect on root growth. The objective of this study was to elucidate the relationship between inhibition of rice root growth induced by high ammonium conditions and ammonium assimilation in the seedlings. Activity of glutamine synthetase (GS) was kept at a low level in the seminal roots of the seedlings grown under high nitrate levels. In contrast, high ammonium levels significantly enhanced the GS activity in the roots, so that Gln abundantly accumulated in the shoots. These results indicate that ammonium assimilation may be activated in the seminal roots under high ammonium conditions. Application of methionine sulfoximine (MSO), an inhibitor of GS, relieved the repression of the seminal root elongation induced by high ammonium concentrations. However, the elongation of lateral roots remained inhibited even under the same condition. Furthermore, MSO drastically increased ammonium level and remarkably decreased Gln level in the shoots grown under high ammonium conditions. These results show that, for rice seedlings, an assimilatory product of ammonium, and not ammonium itself, may serve as an endogenous indicator of the nitrogen status involved in the inhibition of seminal root elongation induced by high levels of exogenous ammonium. PMID- 18419740 TI - Mobility of boron-polyol complexes in the hemiparasitic association between Rhinanthus minor and Hordeum vulgare: the effects of nitrogen nutrition. AB - Boron (B) is an essential nutrient required for plant growth and physiological processes. Long-distance B transport is facilitated by the formation of B-polyol complexes. We investigated B uptake and distribution in response to differing levels of exogenous nitrogen supply in the hemiparasitic association between Rhinanthus minor and Hordeum vulgare (barley) and in unparasitised barley and single Rhinanthus plants. In this system, the polyol mannitol is the major assimilate in Rhinanthus, whereas polyols are not detectable in barley. Furthermore, previous studies have shown that the accumulation of polyols within Rhinanthus is negatively affected by the application of exogenous nitrogen. Within the association, the strongest accumulation of B was detected in lateral buds and inflorescences of Rhinanthus, consistent with the greatest B demand in strong sink organs supplied through the phloem that contain high concentrations of mannitol. In the host, the strongest B accumulation was found in xylem supported leaf lamellae. Roots and sheaths did not accumulate substantial amounts of B, while re-circulation of B through the phloem vessels accounted for only 10% (unparasitised) and 8% (parasitised) of the xylem sap-imported B in the mannitol free barley hosts. In contrast, 53% (attached) and 39% (in the absence of a host) of the xylem sap-imported B was re-circulated in the phloem in the mannitol-rich Rhinanthus. We therefore present the first quantitative uptake and flow models of long-distance B transport in polyol-rich and polyol-free plants. Our findings are consistent with a close relationship between B re-translocation and mannitol concentrations in phloem vessels. PMID- 18419741 TI - Founder haplotype associated with the factor VIII Asp1241Glu polymorphism in a cohort of mild hemophilia A patients. PMID- 18419742 TI - Globular adiponectin induces platelet activation through the collagen receptor GPVI-Fc receptor gamma chain complex. AB - BACKGROUND: The adipocyte-derived cytokine, adiponectin (Ad), exerts potent vascular effects, although the direct effects of Ad on blood platelets are unclear. OBJECTIVE: The influence of globular Ad (gAd) on blood platelet function was investigated. RESEARCH DESIGN AND METHODS: We measured platelet aggregation and tyrosine phosphorylation signaling events in human and mouse platelets. The ability of gAd to activate Glycoprotein VI (GPVI) activity was determined with a NFAT luciferase reporter assay. RESULTS: gAd, but not full length Ad, induced rapid aggregation and granule secretion of human and mouse platelets through a pathway that is ablated under conditions of Src kinase inhibition, indicating a tyrosine kinase-dependent mechanism. Consistent with this, gAd stimulates rapid tyrosine phosphorylation of several proteins in human and mouse platelets. The pattern of increase in tyrosine phosphorylation was similar to that induced by collagen, with the tyrosine kinase Syk and PLCgamma2 being identified among the list of tyrosine phosphorylated proteins. As collagen activates platelet through the GPVI-Fc receptor gamma-chain (FcRgamma) complex, we used FcRgamma null platelets (which also lack GPVI) to explore the mechanism by which gAd stimulates platelets. Stimulation of tyrosine phosphorylation and platelet aggregation by gAd was abolished in FcRgamma null platelets and markedly reduced in the absence of PLCgamma2. Further, GPVI was confirmed as a collagen receptor for gAd by increased luciferase activity in Jurkat T-cells transfected with GPVI. CONCLUSIONS: We identify gAd as a novel ligand for GPVI that stimulates tyrosine kinase-dependent platelet aggregation. Our data raise the possibility that gAd may promote unwanted platelet activation at sites of vascular injury. PMID- 18419743 TI - D-Dimer for venous thromboembolism diagnosis: 20 years later. AB - Twenty years after its first use in the diagnostic workup of suspected venous thromboembolism (VTE), fibrin D-dimer (DD) testing has gained wide acceptance for ruling out this disease. The test is particularly useful in the outpatient population referred to the emergency department because of suspected deep vein thrombosis (DVT) or pulmonary embolism (PE), in which the ruling out capacity concerns every third patient clinically suspected of having the disease. This usefulness is based on the high sensitivity of the test to the presence of VTE, at least for some assays. Due to its poor specificity precluding its use for ruling in VTE, DD testing must be integrated in comprehensive, sequential diagnostic strategies that include clinical probability assessment and imaging techniques such as lower limb venous compression ultrasonography for suspected DVT or multi-slice helical computed tomography for suspected PE. The present narrative review updates the data available on the use of the various commercially available DD assays in the diagnostic approach of clinically suspected VTE in distinct patient populations or situations, including outpatients and inpatients, patients with cancer, older age, pregnancy, a suspected recurrent event, limited thrombus burden, and patients already on anticoagulant treatment. PMID- 18419744 TI - Food-entrained rhythmic expression of PER2 and BMAL1 in murine megakaryocytes does not correlate with circadian rhythms in megakaryopoiesis. AB - BACKGROUND: Circadian rhythms control a vast array of biological processes in a broad spectrum of organisms. The contribution of circadian rhythms to the development of megakaryocytes and the regulation of platelet biology has not been defined. OBJECTIVES: This study tested the hypothesis that murine megakaryocytes exhibit hallmarks of circadian control. METHODS: Mice expressing a PER2::LUCIFERASE circadian reporter protein and C57BI/6 mice were used to establish if megakaryocytes expressed circadian genes in vitro and in vivo. Mice were also subjected to 3 weeks on a restricted feeding regime to separate food entrained from light-entrained circadian rhythms. Quantitative real time polymerase chain reaction (PCR), flow cytometry and imunohistochemistry were employed to analyse gene expression, DNA content and cell-cycle behavior in megakaryocytes collected from mice over a 24-h period. RESULTS: Megakaryocytes exhibited rhythmic expression of the clock genes mPer2 and mBmal1 and circadian rhythms in megakaryopoiesis. mPer2 and mBmal1 expression phase advanced 8 h to coincide with the availability of food; however, food availability had a more complex effect on megakaryopoiesis, leading to a significant overall increase in megakaryocyte ploidy levels and cell-cycle activity. CONCLUSIONS: Normal megakaryopoiesis requires synchrony between food- and light-entrained circadian oscillators. PMID- 18419745 TI - A new VKORC1 allelic variant (p.Trp59Arg) in a patient with partial resistance to acenocoumarol and phenprocoumon. PMID- 18419746 TI - Application of a pharmacogenetic-based warfarin dosing algorithm derived from British patients to predict dose in Swedish patients. PMID- 18419747 TI - Protein S enhances the tissue factor pathway inhibitor inhibition of factor Xa but not its inhibition of factor VIIa-tissue factor. PMID- 18419748 TI - Quercetin induced tissue-type plasminogen activator expression is mediated through Sp1 and p38 mitogen-activated protein kinase in human endothelial cells. AB - BACKGROUND: Wine polyphenol quercetin upregulates tissue-type plasminogen activator (t-PA) transcription in cultured human umbilical cord vein endothelial cells (HUVECs). However, the regulatory elements and signaling pathways involved in this regulation are unknown. OBJECTIVES: We aimed to localize quercetin responsive t-PA promoter elements, identify the proteins that bind these elements, and decipher signaling pathways involved in the regulation of t-PA. METHODS: To localize quercetin-responsive elements, HUVECs were transiently transfected with various t-PA promoter-reporter constructs. Element functionality was evaluated by mutational analysis. Nuclear protein-t-PA element interactions were evaluated by electrophoretic mobility shift assays (EMSAs) and chromatin immunoprecipitation (ChIP) analysis. Mitogen-activated protein kinase (MAPK) inhibitors were used to determine the signaling pathways involved in t-PA regulation. MAPK inhibition effects were evaluated by real-time PCR, immunoblotting analysis, and transfections. Coimmunoprecipitation was used to evaluate MAPK and transcription factor interaction. RESULTS: Deletion of the t-PA promoter region - 288 to - 250 resulted in loss of quercetin responsiveness. This region contains putative Sp1-binding elements, which we termed Sp1a and Sp1b. Sp1b mutation abolished the quercetin-inducible response, whereas Sp1a mutation had no effect. EMSA and ChIP analysis demonstrated quercetin-enhanced Sp1 binding to Sp1b. Inhibition of p38 MAPK abrogated basal and quercetin-induced t-PA expression and promoter activity, as well as quercetin-induced Sp1 binding to Sp1b. Quercetin enhanced p38 MAPK and Sp1 physical association, which was similarly diminished by p38 MAPK inhibition. CONCLUSIONS: We showed, for the first time, the presence of a functional Sp1-binding element in the t-PA promoter controlling quercetin induction via the p38 MAPK pathway. Understanding these mechanisms may provide new insights into polyphenol cardioprotective effects. PMID- 18419749 TI - Ex vivo inhibition of thrombus formation by an anti-glycoprotein VI Fab fragment in non-human primates without modification of glycoprotein VI expression. AB - OBJECTIVES: Glycoprotein (GP)VI is an attractive target for the development of new antithrombotic drugs. Its deficiency protects animals in several models of thrombosis, arterial stenosis and ischemia--reperfusion while inducing no major bleeding tendency. The Fab fragment of one anti-GPVI monoclonal antibody (9O12.2) inhibits all GPVI functions in vitro. The aim of this study was to determine the ex vivo effects of 9O12.2 Fab on hemostasis, coagulation and thrombosis in non human primates. METHODS AND RESULTS: Blood samples were collected from cynomolgus monkeys before and after (30, 90 and 150 min, 1 and 7 days) a bolus injection of 9O12.2 Fab (4 mg kg(-1)) or vehicle. Platelet counts and coagulation tests (prothrombin time, activated partial thromboplastin time) were not modified following Fab injection. The PFA-100 closure time increased during the first hours and returned to initial values on day + 1. Platelet-bound Fab was detected from 30 min to 24 h after Fab injection without GPVI depletion at any time. Collagen-induced platelet aggregation was selectively and fully inhibited at 30 min. Thrombus formation on collagen in flowing whole blood (1500 s(-1)) was delayed and decreased, and collagen-induced or tissue factor-induced thrombin generation in platelet-rich plasma was profoundly inhibited. CONCLUSION: The anti GPVI 9O12.2 Fab inhibits thrombus formation ex vivo in non-human primates with a composite effect on platelet activation and thrombin generation in the absence of GPVI depletion. PMID- 18419750 TI - One fig to bind them all: host conservatism in a fig wasp community unraveled by cospeciation analyses among pollinating and nonpollinating fig wasps. AB - The study of chalcid wasps that live within syconia of fig trees (Moraceae, Ficus), provides a unique opportunity to investigate the evolution of specialized communities of insects. By conducting cospeciation analyses between figs of section Galoglychia and some of their associated fig wasps, we show that, although host switches and duplication have evidently played a role in the construction of the current associations, the global picture is one of significant cospeciation throughout the evolution of these communities. Contrary to common belief, nonpollinating wasps are at least as constrained as pollinators by their host association in their diversification in this section. By adapting a randomization test in a supertree context, we further confirm that wasp phylogenies are significantly congruent with each other, and build a "wasp community" supertree that retrieves Galoglychia taxonomic subdivisions. Altogether, these results probably reflect wasp host specialization but also, to some extent, they might indicate that niche saturation within the fig prevents recurrent intrahost speciation and host switching. Finally, a comparison of ITS2 sequence divergence of cospeciating pairs of wasps suggests that the diversification of some pollinating and nonpollinating wasps of Galoglychia figs has been synchronous but that pollinating wasps exhibit a higher rate of molecular evolution. PMID- 18419751 TI - Socially acquired host-specific mimicry and the evolution of host races in Horsfield's bronze-cuckoo Chalcites basalis. AB - Coevolution between parasites and their hosts typically leads to increasing specialization on host species by the parasite. Where multiple hosts are parasitized, specialization on each host can result in genetic divergence within the parasite population to create host races, and, ultimately, new species. We investigate how host-specific traits arise in Horsfield's bronze-cuckoo Chalcites basalis nestlings. Newly hatched cuckoos evict host young from the nest, yet in the absence of a model they accurately mimic the different begging calls of a primary host (superb fairy-wren, Malurus cyaneus) and a secondary host (buff rumped thornbill, Acanthiza reguloides). Using cross-fostering experiments, we show that begging calls are modified after parasitism, through experience. Further, we demonstrate the mechanism by which mimetic calls are acquired. All cuckoo nestlings initially produced the call of their primary host. When cross fostered as eggs to a secondary host, calls increased in variability and were rapidly modified to resemble those of the secondary host through shaping by host parents. We suggest that plasticity in the development of host-specific traits after parasitism is likely to reduce selection for host race formation. PMID- 18419752 TI - Natural selection on floral traits through male and female function in wild populations of the heterostylous daffodil Narcissus triandrus. AB - Measurements of natural selection in hermaphrodite populations require the analysis of performance through both female and male sex functions. Here, we investigate selection on three floral traits: flower number, flower length, and corona width through both sex functions in natural populations of the tristylous daffodil Narcissus triandrus. Selection through female function was examined in six populations, and in two of these we also estimated male selection gradients using multilocus microsatellite genotyping of parents and offspring. We detected significant directional selection for flower number through female function, and significant stabilizing selection for corona width and flower length through male function. Variation in male reproductive success was strongly influenced by the distance between mates and was significantly higher than variation in female reproductive success in one population, a result consistent with Bateman's principle. However, variation through both sex functions was similar in the other population and there was a significant negative correlation between female and male fitness indicating sex-specific trade-offs in reproductive success. Selection on floral design in N. triandrus was stronger through male than female function probably because floral morphology plays an important role in promoting effective cross-pollen transfer in populations of this heterostylous species. PMID- 18419753 TI - TrkA receptor endolysosomal degradation is both ubiquitin and proteasome dependent. AB - Gaps in our knowledge exist regarding the degradation of the tropomyosin regulated kinase A (TrkA) receptor after addition of neurotrophin, nerve growth factor (NGF). TrkA is rapidly and transiently ubiquitinated upon addition of NGF. Here, we demonstrate that the polyubiquitin tag plays a definitive role in receptor sorting. Treatment of PC12 cells with lactacystin prevented NGF dependent deubiquitination and degradation of TrkA. However, treatment with methylamine, bafilomycin or leupeptin, did not prevent NGF-dependent deubiquitination but blocked the degradation of TrkA. Employing co immunoprecipitation, biochemical fractionation and confocal microscopy, the kinetics of receptor trafficking post-internalization was observed to occur as a sequel from endosome/multivesicular body, proteasomes, culminating with degradation in the lysosomes. The trafficking of the polyubiquitin-deficient TrkA receptor mutant K485R was impaired and likewise failed to degrade revealing that the receptor escapes degradation. The interaction of TrkA with proteasomes was confirmed by purification and co-immunoprecipitation. We provide evidence that proteasomal deubiquitinating enzymes trim K63-ubiquitin chains from the TrkA receptor prior to its delivery to lysosomes for degradation. Taken together, our results reveal the existence of a novel proteasome-dependent step in receptor degradation. PMID- 18419755 TI - A direct role for phosphatidylinositol-4,5-bisphosphate in unconventional secretion of fibroblast growth factor 2. AB - Fibroblast growth factor 2 (FGF-2) is a mitogen that is exported from cells by an endoplasmic reticulum/Golgi-independent secretory pathway. Recent findings have shown that FGF-2 export occurs by direct translocation from the cytoplasm across the plasma membrane into the extracellular space. Here, we report that FGF-2 contains a binding site for phosphatidylinositol-4,5-bisphosphate [PI(4,5)P(2)], the principal phosphoinositide species associated with plasma membranes. Intriguingly, in the context of a lipid bilayer, the interaction between FGF-2 and PI(4,5)P(2) is shown to depend on a lipid background that resembles plasma membranes. We show that the interaction with PI(4,5)P(2) is critically important for FGF-2 secretion as experimental conditions reducing cellular levels of PI(4,5)P(2) resulted in a substantial drop in FGF-2 export efficiency. Likewise, we have identified FGF-2 variant forms deficient for binding to PI(4,5)P(2) that were found to be severely impaired with regard to export efficiency. These data show that a transient interaction with PI(4,5)P(2) associated with the inner leaflet of plasma membranes represents the initial step of the unconventional secretory pathway of FGF-2. PMID- 18419754 TI - The novel sorting nexin SNX33 interferes with cellular PrP formation by modulation of PrP shedding. AB - The cellular prion protein (PrP(c)) is a glycosyl-phosphatidylinositol (GPI) anchored protein trafficking in the secretory and endocytic pathway and localized mainly at the plasma membrane. Conversion of PrP(c) into its pathogenic isoform PrP(Sc) is associated with pathogenesis and transmission of prion diseases. Intramolecular cleavage in the middle, the extreme C-terminal part or within the GPI anchor and shedding of PrP(c) modulate this conversion process by reducing the substrate for prion formation. These phenomena provide similarities with the processing of amyloid precursor protein in Alzheimer's disease. Sorting nexins are a family of proteins with important functions in protein trafficking. In this study, we investigated the role of the newly described sorting nexin 33 (SNX33) in trafficking and processing of PrP(c). We found that overexpression of SNX33 in neuronal and non-neuronal cell lines resulted in increased shedding of full length PrP(c) from the plasma membrane and modulated the rate of PrP(c) endocytosis. This was paralleled by reduction of PrP(Sc) formation in persistently and newly infected cells. Using deletion mutants, we demonstrate that production of PrP fragment N1 is not influenced by SNX33. Our data provide new insights into the cellular mechanisms of PrP(c) shedding and show how this can affect cellular PrP(Sc) conversion. PMID- 18419759 TI - Spinal CCL2 pronociceptive action is no longer effective in CCR2 receptor antagonist-treated rats. AB - A better understanding of the mechanisms linked to chemokine pronociceptive effects is essential for the development of new strategies to better prevent and treat chronic pain. Among chemokines, MCP-1/CCL2 involvement in neuropathic pain processing is now established. However, the mechanisms by which MCP-1/CCL2 exerts its pronociceptive effects are still poorly understood. In the present study, we demonstrate that MCP-1/CCL2 can alter pain neurotransmission in healthy rats. Using immunohistochemical studies, we first show that CCL2 is constitutively expressed by primary afferent neurons and their processes in the dorsal horn of the spinal cord. We also observe that CCL2 is co-localized with pain-related peptides (SP and CGRP) and capsaicin receptor (VR1). Accordingly, using in vitro superfusion system of lumbar dorsal root ganglion and spinal cord explants of healthy rats, we show that potassium or capsaicin evoke calcium-dependent release of CCL2. In vivo, we demonstrate that intrathecal administration of CCL2 to healthy rats produces both thermal hyperalgesia and sustained mechanical allodynia (up to four consecutive days). These pronociceptive effects of CCL2 are completely prevented by the selective CCR2 antagonist (INCB3344), indicating that CCL2-induced pain facilitation is elicited via direct spinal activation of CCR2 receptor. Therefore, preventing the activation of CCR2 might provide a fruitful strategy for treating pain. PMID- 18419760 TI - Glutamate activates GFAP gene promoter from cultured astrocytes through TGF-beta1 pathways. AB - Glial cells are currently viewed as active partners of neurons in synapse formation. The close proximity of astrocytes to the synaptic cleft suggests that these cells might be potential targets for neuronal-released molecules although this issue has been less addressed. Here, we evaluated the role of the excitatory neurotransmitter, glutamate, in astrocyte differentiation. We recently demonstrated that cortical neurons activate the gene promoter of the astrocyte maturation marker, GFAP (glial fibrillary acidic protein) of cerebral cortex astrocytes by inducing TGF-beta1 (transforming growth factor beta 1) secretion in vitro. To access the effect of glutamate on GFAP gene, we used transgenic mice bearing the beta-Galactosidase (beta-Gal) reporter gene under the regulation of the GFAP gene promoter. We report that 100 muM glutamate activates the GFAP gene promoter of astrocytes from cerebral cortex revealed by a significant increase in the number of beta-Gal positive astrocytes. Neutralizing antibodies against TGF beta completely prevented glutamate and neuronal-induction of GFAP gene, thus indicating that this event is mediated by TGF-beta1. Further, induction of GFAP gene in response to glutamate was followed by nuclear translocation of the Smad transcription factor, a hallmark of TGF-beta1 pathway activation. The antagonist of the metabotropic glutamate receptor, MCPG, inhibited neuronal effect suggesting that neuronal activation of GFAP gene promoter involves glutamate metabotropic receptors. MAPK (PD98059) and PI3K (LY294002) inhibitors fully prevented activation of GFAP gene promoter by all treatments. Surprisingly, these inhibitors also abrogated TGF-beta1 direct action on GFAP gene although they did not inhibit Smad-2 phosphorylation, suggesting that TGF-beta1-induced GFAP gene activation might involve cooperation between the canonical and non-canonical TGF beta pathways. Together, our results suggest that glial metabotropic glutamate 2/3 receptor activation by neurons induces TGF-beta1 secretion, leading to GFAP gene activation and astrocyte differentiation and involves Smad and MAPK/PI3K pathways. Our work provides evidence that astrocytes surrounding synapses are target of neuronal activity and might shed light into the role of glial cells into neurological disorders associated with glutamate neurotoxicity. PMID- 18419761 TI - Orphan nuclear receptor, Nurr-77 was a possible target gene of butylidenephthalide chemotherapy on glioblastoma multiform brain tumor. AB - The natural compound n-butylidenephthalide (BP), which is isolated from the chloroform extract of Angelica sinensis, has been investigated for its antitumoral effects on glioblastoma multiform (GBM) brain tumors both in vitro and in vivo. To determine the mechanism of BP-induced growth arrest and apoptosis, we examined BP-induced changes in gene expression by microarray screening using human GBM brain tumor cells. This analysis identified several BP inducible genes, including the nuclear receptors NOR-1, Nurr1, and Nur77. Among these genes, Nur77 is particularly interesting because it plays an important role in the apoptotic processes in various tumor cell lines. BP was able to increase Nur77 mRNA and protein expression in a time-dependent manner. After BP treatment in GBM 8401 cells, Nur77 translocated from the nucleus to the cytoplasm, the cytochrome c was released from the mitochondria, and caspase 3 became activated. Furthermore, using Nur77 promoter-luciferase assay, BP increased Nur77 was AP1 related. Inhibition of BP-induced Nur77 expression by Nur77 short interfering RNA blocked BP-induced apoptosis in GBM 8401 cells, suggesting that the induction of Nur77 negatively affected GBM 8401 cell survival. In summary, our results suggest that up-regulation of Nur77 may explain the antitumoral activity of BP in brain tumor cells. PMID- 18419762 TI - Post-endocytic fates of delta-opioid receptor are regulated by GRK2-mediated receptor phosphorylation and distinct beta-arrestin isoforms. AB - Once internalized, some G protein-coupled receptors (GPCRs) can recycle back to the cell surface, while some of them are delivered to lysosomes for degradation. Because recycling and degradation represent two opposing receptor fates, understanding the mechanisms that determine post-endocytic fate of GPCRs is of great importance. Our recent work has verified that agonist-induced internalization of delta-opioid receptor (DOR) employs both phosphorylation dependent and -independent mechanisms in HEK293 cells. To investigate whether these two internalization mechanisms work differently in receptor regulation, we monitored receptor post-endocytic fates using flow cytometry, surface receptor biotinylation and radioligand binding assays. Results showed that the internalized wild type DOR could either recycle to the cell surface or be degraded. Mutant DOR M4/5/6, which lacks all three G protein-coupled receptor kinase 2 (GRK2) phosphorylation sites, could also internalize upon agonist challenge although in a reduced level as compared with the wild type counterpart. However, the internalized mutant DOR could not recycle back to the cell surface and all mutant DOR was degraded after internalization. Inhibition of GRK2 expression by GRK2 RNAi also strongly attenuated recycling of DOR. Furthermore, overexpression of GRK2, which significantly increased receptor phosphorylation and internalization, also targeted more internalized receptors to the recycling pathway. These data suggest that GRK2-catalyzed receptor phosphorylation is critically involved in DOR internalization and recycling, and the phosphorylation independent internalization leads to receptor degradation. Data obtained from beta-arrestin1 and beta-arrestin2 RNAi experiments indicated that both beta arrestin1 and beta-arrestin2 participate in phosphorylation-dependent internalization and the subsequent recycling of DOR. However, phosphorylation independent internalization and degradation of DOR were strongly blocked by beta arrestin2 RNAi, but not beta-arrestin1 RNAi. Taken together, these data demonstrate for the first time that GRK2 phosphorylation-dependent internalization mediated by both beta-arrestin1 and beta-arrestin2 leads DOR to recycle, whereas GRK2-independent internalization mediated by beta-arrestin2 alone leads to receptor degradation. Thus, the post-endocytic fate of internalized DOR can be regulated by GRK2-catalyzed receptor phosphorylation as well as distinct beta-arrestin isoforms. PMID- 18419763 TI - Inhibition of MMP-3 or -9 suppresses lipopolysaccharide-induced expression of proinflammatory cytokines and iNOS in microglia. AB - Recently, matrix metalloproteinases (MMPs) are emerging as important molecules in neuroinflammation as well as neuronal cell death. However, the role of MMPs in activated microglia remains unclear. In the present study, we found that expressions of MMP-1, -3, -8 and -9 were significantly induced by single or combined treatment of immunostimulants lipopolysaccharide (LPS) or phorbol myristate acetate (PMA) in primary cultured microglia and BV2 microglial cells. Inhibition of MMP-3 or -9 significantly suppressed the expression of iNOS and pro inflammatory cytokines and the activities of NF-kappaB, AP-1, and MAPK in LPS stimulated microglia. The results suggest that MMP-3 and -9 both mediate LPS induced inflammatory reactions. Inhibition of reactive oxygen species (ROS) by N acetyl-cysteine or diphenylene iodonium significantly suppressed the expression of MMP-3, MMP-9, NO and TNF-alpha in LPS-stimulated microglia, suggesting that ROS is an early signaling inducer in LPS-stimulated microglial cells. MMP inhibitors also suppressed ROS production, suggesting a cross-talk between ROS and MMPs. Collectively, the present study demonstrates that MMP-3 and MMP-9 play a role as inflammatory mediators in activated microglia. Pharmacological intervention of MMPs especially MMP-3 and -9 would be a therapeutic strategy for the treatment of inflammatory diseases in the CNS caused by over-activation of microglial cells. PMID- 18419764 TI - Pin1 promotes cell death in NGF-dependent neurons through a mechanism requiring c Jun activity. AB - Developing neurons deprived of trophic support undergo apoptosis mediated by activation of c-Jun N-terminal kinases (JNK) and c-Jun, induction of the Bcl-2 homology 3-only protein Bim(EL), Bax-dependent loss of mitochondrial cytochrome c, and caspase activation. However, the mechanisms that regulate each of these events are only partially understood. Here we show that the prolyl isomerase Pin1 functions as a positive regulator of neuronal death through a c-Jun-dependent mechanism. Ectopic Pin1 promoted caspase-dependent death of NGF-maintained neurons that was associated with an accumulation of Ser(63)-phosphorylated c-Jun in neuronal nuclei and was partially dependent on Bax. Downregulating Pin1 prior to NGF withdrawal suppressed the accumulation of phosphorylated c-Jun, inhibited the release of cytochrome c, and significantly delayed cell death. Pin1 knockdown inhibited NGF deprivation-induced death to a similar extent in Bim (+/+) and Bim (-/-) neurons. The protective effect of Pin1 knockdown was significantly greater than that caused by loss of Bim and nearly identical to that caused by a dominant negative form of c-Jun. Finally, cell death induced by ectopic Pin1 was largely blocked by expression of dominant negative c-Jun. These results suggest a novel mechanism by which Pin1 promotes cell death involving activation of c-Jun. PMID- 18419765 TI - Myelin-induced microglial neurotoxicity can be controlled by microglial metabotropic glutamate receptors. AB - Microglia are present in an activated state in multiple sclerosis lesions. Incubation of primary cultured rat microglia with rat-brain derived myelin (0.1-1 microg/mL) for 24 h induced microglial activation; cells displayed enhanced ED1 staining, expression of inducible nitric oxide synthase, production and release of the cytokine tumour necrosis factor-alpha and glutamate release. Exposure of microglia to myelin induced the expression of neuronal caspases and ultimately neuronal death in cultured cerebellar granule cell neurons; neurotoxicity was directly because of microglial-derived soluble toxins. Co-incubation of microglia with agonists or antagonists of different metabotropic glutamate receptor (mGluR) subtypes ameliorated microglial neurotoxicity by inhibiting soluble neurotoxin production. Activation of microglial mGluR2 exacerbated myelin-evoked neurotoxicity whilst activation of mGluR3 was protective as was activation of group III mGluRs. These data show that myelin-induced microglial neurotoxicity can be prevented by regulation of mGluRs and suggest these receptors on microglia may be promising targets for therapeutic intervention in multiple sclerosis. PMID- 18419766 TI - Ethanol mimics ligand-mediated activation and endocytosis of IL-1RI/TLR4 receptors via lipid rafts caveolae in astroglial cells. AB - We have recently reported that ethanol-induced inflammatory processes in the brain and glial cells are mediated via the activation of interleukin-1 beta receptor type I (IL-1RI)/toll-like receptor type 4 (TLR4) signalling. The mechanism(s) by which ethanol activates these receptors in astroglial cells remains unknown. Recently, plasma membrane microdomains, lipid rafts, have been identified as platforms for receptor signalling and, in astrocytes, rafts/caveolae constitute an important integrators of signal events and trafficking. Here we show that stimulation of astrocytes with IL-1beta, lipopolysaccharide or ethanol (10 and 50 mM), triggers the translocation of IL 1RI and/or TLR4 into lipid rafts caveolae-enriched fractions, promoting the recruitment of signalling molecules (phospho-IL-1R-associated kinase and phospho extracellular regulated-kinase) into these microdomains. With confocal microscopy, we further demonstrate that IL-1RI is internalized by caveolar endocytosis via enlarged caveosomes organelles upon IL-1beta or ethanol treatment, which sorted their IL-1RI cargo into the endoplasmic reticulum-Golgi compartment and into the nucleus of astrocytes. In short, our findings demonstrate that rafts/caveolae are critical for IL-1RI and TLR4 signalling in astrocytes, and reveal a novel mechanism by which ethanol, by interacting with lipid rafts caveolae, promotes IL-1RI and TLR4 receptors recruitment, triggering their endocytosis via caveosomes and downstream signalling stimulation. These results suggest that TLRs receptors are important targets of ethanol-induced inflammatory damage in the brain. PMID- 18419767 TI - Histamine induces neural stem cell proliferation and neuronal differentiation by activation of distinct histamine receptors. AB - Histamine has neurotransmitter/neuromodulator functions in the adult brain, but its role during CNS development has been elusive. We studied histamine effects on proliferation, cell death and differentiation of neuroepithelial stem cells from rat cerebral cortex in vitro. RT-PCR and Western blot experiments showed that proliferating and differentiated cells express histamine H(1), H(2) and H(3) receptors. Treatments with histamine concentrations (100 nM-1 mM) caused significant increases in cell numbers without affecting Nestin expression. Cell proliferation was evaluated by BrdU incorporation; histamine caused a significant increase dependent on H(2) receptor activation. Apoptotic cell death during proliferation was significantly decreased at all histamine concentrations, and cell death was promoted in a concentration-dependent manner by histamine in differentiated cells. Immunocytochemistry studies showed that histamine increased 3-fold the number of neurons after differentiation, mainly by activation of H(1) receptor, and also significantly decreased the glial (astrocytic) cell proportion, when compared to control conditions. In summary, histamine increases cell number during proliferative conditions, and has a neuronal-differentiating action on neural stem cells, suggesting that the elevated histamine concentration reported during development might play a role in cerebrocortical neurogenesis, by activation of H(2) receptors to promote proliferation of neural precursors, and favoring neuronal fate by H(1)-mediated stimulation. PMID- 18419768 TI - Tetrahydrobiopterin shows chaperone activity for tyrosine hydroxylase. AB - Tyrosine hydroxylase (TH) is the rate-limiting enzyme in the synthesis of catecholamine neurotransmitters. Primary inherited defects in TH have been associated with l-DOPA responsive and non-responsive dystonia and infantile parkinsonism. In this study, we show that both the cofactor (6R)-l-erythro 5,6,7,8-tetrahydrobiopterin (BH(4)) and the feedback inhibitor and catecholamine product dopamine increase the kinetic stability of human TH isoform 1 in vitro. Activity measurements and synthesis of the enzyme by in vitro transcription translation revealed a complex regulation by the cofactor including both enzyme inactivation and conformational stabilization. Oral BH(4) supplementation to mice increased TH activity and protein levels in brain extracts, while the Th-mRNA level was not affected. All together our results indicate that the molecular mechanisms for the stabilization are a primary folding-aid effect of BH(4) and a secondary effect by increased synthesis and binding of catecholamine ligands. Our results also establish that orally administered BH(4) crosses the blood-brain barrier and therapeutic regimes based on BH(4) supplementation should thus consider the effect on TH. Furthermore, BH(4) supplementation arises as a putative therapeutic agent in the treatment of brain disorders associated with TH misfolding, such as for the human TH isoform 1 mutation L205P. PMID- 18419769 TI - Brain metabolism in adult chronic hydrocephalus. AB - Normal pressure hydrocephalus (NPH) is the most frequent form of chronic hydrocephalus in adults. NPH remains underdiagnosed although between 5% and 10% of all demented patients may suffer from this disorder. As dementia is an increasing demographic problem, treatable forms such as in NPH have become a central issue in neurology. Despite the traditional perception of hydrocephalus being a disorder of disturbed CSF dynamics, in NPH metabolic impairment seems at least as important. So far, the only valid animal model of NPH is chronic adult kaolin hydrocephalus. In this model, opening of alternative CSF outflow pathways leads to normal or near-normal intracranial pressure and CSF outflow resistance. Yet, various metabolic disturbances cause ongoing ventricular enlargement and characteristic symptoms including cognitive decline and gait ataxia. Delayed hippocampal neuronal death, accumulation of beta-amyloid and disturbed cholinergic neurotransmission may contribute to memory dysfunction. Compromised periventricular blood flow, decreased dopamine levels in the substantia nigra and damaged striatal GABAergic interneurons may reflect basal ganglia symptoms. At least in human hydrocephalus cerebrovascular co-morbidity of the white matter plays an important role as well. It seems that in hydrocephalus from a certain 'point of no return' metabolic impairment becomes decoupled from CSF dynamics and, at least partly, self-sustained. This is probably the reason why despite restored CSF circulation by shunting many patients with chronic hydrocephalus still suffer from severe neurological deficits. The present paper offers a comprehensive review of the experimental and clinical data suggesting metabolic disturbances in chronic hydrocephalus. PMID- 18419770 TI - Characterization of OPA1 isoforms isolated from mouse tissues. AB - OPA1, a nuclear encoded mitochondrial protein causing autosomal dominant optic atrophy, is a key player in mitochondrial fusion and cristae morphology regulation. In the present study, we have compared the OPA1 transcription and translation products of different mouse tissues. Unlike in humans, we found only two exons (4b and 5b) to be involved in alternative splicing. The relative abundance of the resulting four different splice variants is tissue-dependent. Proteolytic cleavage by mitochondrial processing peptidase generates two long forms, isoforms 1 and 7, which lead to three short forms representing the end products after further proteolytic processing. In contrast, isoforms 5 and 8 are directly processed into their corresponding short forms. Short form 1 molecules form 184 kDa dimers, whereas all other isoforms contribute to 285 kDa complexes. Coiled-coil domains of the OPA1 protein specifically homo-associate and may be involved in the formation of these complexes. Furthermore, the region encoded by exon 5b inhibits the self-association of coiled-coil domain-I. Finally, our data pinpoint isoform 1 as the, by far, most abundant isoform in the nervous tissue. We postulate that manipulation of isoform 1 protein levels in relation to the other isoforms induces changes in the mitochondrial network in the cell and therefore, mutations affecting the level of functional isoform 1 could lead to devastating effects on retinal ganglion cells. PMID- 18419771 TI - Expression and processing of Plasmodium berghei SERA3 during liver stages. AB - Cysteine proteases mediate liberation of Plasmodium berghei merozoites from infected hepatocytes. In an attempt to identify the responsible parasite proteases, we screened the genome of P. berghei for cysteine protease-encoding genes. RT-PCR analyses revealed that transcription of four out of five P. berghei serine repeat antigen (PbSERA) genes was strongly upregulated in late liver stages briefly before the parasitophorous vacuole membrane ruptured to release merozoites into the host cell cytoplasm, suggesting a role of PbSERA proteases in these processes. In order to characterize PbSERA3 processing, we raised an antiserum against a non-conserved region of the protein and generated a transgenic P. berghei strain expressing a TAP-tagged PbSERA3 under the control of the endogenous promoter. Immunofluorescence assays revealed that PbSERA3 leaks into the host cell cytoplasm during merozoite development, where it might contribute to host cell death or activate host cell proteases that execute cell death. Importantly, processed PbSERA3 has been detected by Western blot analysis in cell extracts of schizont-infected cells and merozoite-infected detached hepatic cells. PMID- 18419772 TI - Mycobacterium tuberculosis heat shock protein 60 modulates immune response to PPD by manipulating the surface expression of TLR2 on macrophages. AB - The T-helper (Th) 1 T-cell response to purified protein derivative (PPD) is known to be suppressed in tuberculosis patients which favours intracellular survival of the bacilli. We demonstrate that the Mycobacterium tuberculosis heat shock protein 60 (Mtbhsp60) plays an important role to skew the anti-PPD T-cell response towards the Th2 type when macrophages were used as antigen presenting cells. We found that the PPD-induced IL-12 p40 was downregulated in macrophages by Mtbhsp60. The Mtbhsp60 preferentially induced Toll-like receptor (TLR) 2 without affecting TLR4 expression on macrophages. Interaction of Mtbhsp60 with TLR2 resulted in significant suppression of nuclear c-rel and consequently IL-12 p40 levels in PPD-activated macrophages. Our findings reveal a unique role of the Mtbhsp60 favouring development of Th2 type response by upregulating surface expression of TLR2 on macrophages which could be a survival strategy adopted by the bacilli. PMID- 18419773 TI - Vesicular transport in Histoplasma capsulatum: an effective mechanism for trans cell wall transfer of proteins and lipids in ascomycetes. AB - Vesicular secretion of macromolecules has recently been described in the basidiomycete Cryptococcus neoformans, raising the question as to whether ascomycetes similarly utilize vesicles for transport. In the present study, we examine whether the clinically important ascomycete Histoplasma capsulatum produce vesicles and utilized these structures to secrete macromolecules. Transmission electron microscopy (TEM) shows transcellular secretion of vesicles by yeast cells. Proteomic and lipidomic analyses of vesicles isolated from culture supernatants reveal a rich collection of macromolecules involved in diverse processes, including metabolism, cell recycling, signalling and virulence. The results demonstrate that H. capsulatum can utilize a trans-cell wall vesicular transport secretory mechanism to promote virulence. Additionally, TEM of supernatants collected from Candida albicans, Candida parapsilosis, Sporothrix schenckii and Saccharomyces cerevisiae documents that vesicles are similarly produced by additional ascomycetes. The vesicles from H. capsulatum react with immune serum from patients with histoplasmosis, providing an association of the vesicular products with pathogenesis. The findings support the proposal that vesicular secretion is a general mechanism in fungi for the transport of macromolecules related to virulence and that this process could be a target for novel therapeutics. PMID- 18419774 TI - Chemotaxis of Entamoeba histolytica towards the pro-inflammatory cytokine TNF is based on PI3K signalling, cytoskeleton reorganization and the Galactose/N acetylgalactosamine lectin activity. AB - Entamoeba histolytica is the protozoan parasite responsible for human amoebiasis. During invasive amoebiasis, migration is an essential process and it has previously been shown that the pro-inflammatory compound tumour necrosis factor (TNF) is produced and that it has a migratory effect on E. histolytica. This paper focuses on the analysis of parasite signalling and cytoskeleton changes leading to directional motility. TNF-induced signalling was PI3K-dependent and could lead to modifications in the polarization of certain cytoskeleton-related proteins. To analyse the effect of TNF signalling on gene expression, we used microarray analysis to screen for genes encoding proteins that were potentially important during chemotaxis towards TNF. Interestingly, we found that elements of the galactose/N-acetylgalactosamine lectin (Gal/GalNAc lectin) were upregulated during chemotaxis as well as genes encoding proteins involved in cytoskeleton dynamics. The alpha-actinin protein appeared to be an important candidate to link the Gal/GalNAc lectin to the cytoskeleton during chemotaxis signalling. Dominant negative parasites blocked for Gal/GalNAc lectin signalling were no longer able to chemotax towards TNF. These results have given us an insight on how E. histolytica changes its cytoskeleton dynamics during chemotaxis and revealed the capital role of PI3K and Gal/GalNAc lectin signalling in chemotaxis. PMID- 18419776 TI - Left ventricular function in the post-infarct failing mouse heart by magnetic resonance imaging and conductance catheter: a comparative analysis. AB - AIM: Murine myocardial infarction (MI) models are increasingly used in heart failure studies. Magnetic resonance imaging (MRI) and pressure-volume loops by conductance catheter (CC) enable physiological phenotyping. We performed a comparative analysis of MRI vs. CC to assess left ventricular (LV) function in the failing mouse heart. METHODS: MI was created by LAD ligation. MRI (day 14) and CC (day 15) were used to determine LV end-diastolic volume (EDV), end systolic volume (ESV) and ejection fraction (EF). RESULTS: Pooled data yielded moderate-to-strong linear correlations: EDV: R = 0.61; ESV: R = 0.72; EF: R = 0.81. We analysed three groups, no MI (sham, n = 10), small MI (<30% of LV, n = 14) and large MI (>30%, n = 20). Volumes and EF were consistently lower by CC than by MRI, but group differences were evident for both techniques. Receiver operating characteristic analysis indicated good sensitivity and specificity for both techniques, with superior results for MRI. CONCLUSIONS: CC and MRI are highly valuable for evaluation of LV volume and function. MRI is recommended for longitudinal studies, accurate absolute volumes and anatomical information. Unique features of CC are its online signal with high temporal resolution, and advanced analysis of LV function and energetics. PMID- 18419775 TI - Conserved and non-conserved features of HIV-1 and SIVagm Vif mediated suppression of APOBEC3 cytidine deaminases. AB - Human cytidine deaminase APOBEC3C (A3C) acts as a potent inhibitor of SIVagm and can be regulated by both HIV-1 and SIVagm Vif. The mechanism by which Vif suppresses A3C is unknown. In the present study, we demonstrate that both HIV-1 and SIVagm Vif can act in a proteasome-dependent manner to overcome A3C. SIVagm Vif requires the Cullin5-ElonginB-ElonginC E3 ubiquitin ligase for the degradation of A3C as well as the suppression of its antiviral activity. Mutation of a residue critical for the species-specific recognition of human or monkey A3G by HIV-1 Vif or SIVagm Vif in A3C had little effect on HIV-1 or SIVagm Vif mediated degradation of A3C. Although the amino-terminal region of A3G was not important for Vif-mediated degradation, the corresponding region in A3C was critical. A3C mutants that were competent for Vif binding but resistant to Vif mediated degradation were identified. These data suggest that primate lentiviral Vif molecules have evolved to recognize multiple host APOBEC3 proteins through distinct mechanisms. However, Cul5-E3 ubiquitin ligase appears to be a common pathway hijacked by HIV-1 and SIV Vif to defeat APOBEC3 proteins. Furthermore, Vif and APOBEC3 binding is not sufficient for target protein degradation indicating an important but uncharacterized Vif function. PMID- 18419777 TI - Physiological consequences of the P2328S mutation in the ryanodine receptor (RyR2) gene in genetically modified murine hearts. AB - AIM: To explore the physiological consequences of the ryanodine receptor (RyR2) P2328S mutation associated with catecholaminergic polymorphic ventricular tachycardia (CPVT). METHODS: We generated heterozygotic (RyR2 p/s) and homozygotic (RyR2 s/s) transgenic mice and studied Ca2+ signals from regularly stimulated, Fluo-3-loaded, cardiac myocytes. Results were compared with monophasic action potentials (MAPs) in Langendorff-perfused hearts under both regular and programmed electrical stimulation (PES). RESULTS: Evoked Ca2+ transients from wild-type (WT), heterozygote (RyR2 p/s) and homozygote (RyR2 s/s) myocytes had indistinguishable peak amplitudes with RyR2 s/s showing subsidiary events. Adding 100 nm isoproterenol produced both ectopic peaks and subsidiary events in WT but not RyR2 p/s and ectopic peaks and reduced amplitudes of evoked peaks in RyR2 s/s. Regularly stimulated WT, RyR2 p/s and RyR2 s/s hearts showed indistinguishable MAP durations and refractory periods. RyR2 p/s hearts showed non-sustained ventricular tachycardias (nsVTs) only with PES. Both nsVTs and sustained VTs (sVTs) occurred with regular stimuli and PES with isoproterenol treatment. RyR2 s/s hearts showed higher incidences of nsVTs before but mainly sVTs after introduction of isoproterenol with both regular stimuli and PES, particularly at higher pacing frequencies. Additionally, intrinsically beating RyR2 s/s showed extrasystolic events often followed by spontaneous sVT. CONCLUSION: The RyR2-P2328S mutation results in marked alterations in cellular Ca2+ homeostasis and arrhythmogenic properties resembling CPVT with greater effects in the homozygote than the heterozygote demonstrating an important gene dosage effect. PMID- 18419778 TI - Regression of left ventricular hypertrophy and aortic remodelling in NO-deficient hypertensive rats: effect of L-arginine and spironolactone. AB - AIM: We investigated, whether the substrate for nitric oxide (NO) formation -L arginine - and the aldosterone receptor antagonist - spironolactone - are able to reverse alterations of the left ventricle (LV) and aorta in N(omega)-nitro-L arginine methyl ester (L-NAME)-induced hypertension. METHODS: Six groups of male adult Wistar rats were investigated: controls after 4 and 7 weeks of experiment, rats treated with L-NAME for 4 weeks and three recovery groups: spontaneous reversion (4 weeks L-NAME + 3 weeks placebo), spironolactone-induced reversion (4 weeks L-NAME + 3 weeks spironolactone) and L-arginine-induced reversion (4 weeks L-NAME+ 3 weeks L-arginine). Blood pressure was measured by tail-cuff plethysmography. Relative weight of the LV, myocardial fibrosis (based upon histomorphometry and hydroxyproline determination) and conjugated dienes in the LV and aortic cross-sectional area, inner diameter and wall thickness were determined. NO-synthase activity was investigated in the LV and aorta. RESULTS: L NAME administration induced hypertension, left ventricular hypertrophy (LVH), LV fibrosis, aortic thickening and diminution of NO-synthase activity in the LV and aorta. Reduction in blood pressure and regression of LVH were observed in all recovery groups, yet reduction in LV fibrosis and aortic thickening were not. NO synthase activity was restored only in the L-arginine and spironolactone group. CONCLUSION: In our study, the reversion of hypertension and LVH was not dependent on the restoration of NO-synthase activity. Moreover, LV fibrosis and aortic remodelling seem to be more resistant to conditions resulting in regression of LVH. Preserved level of fibrosis in the initial period of LVH regression might result in loss of structural homogeneity and possible functional alterations of the LV. PMID- 18419779 TI - Glucan, water dikinase phosphorylates crystalline maltodextrins and thereby initiates solubilization. AB - Starch phosphorylation by glucan, water dikinase (GWD; EC 2.7.9.4) is an essential step in the breakdown of native starch particles, but the underlying mechanisms have remained obscure. In this paper, the initial reactions of starch degradation were analyzed using crystallized maltodextrins as model carbohydrates. As revealed by X-ray diffraction analysis, the crystallized maltodextrins represent the B-type starch allomorph. Recombinant GWD phosphorylated crystalline maltodextrins with a high specific activity (55-60 nmol mg-1 protein min-1), but exhibited very little activity with the same maltodextrins that had been solubilized by heat treatment. Recombinant phosphoglucan, water dikinase (PWD; EC 2.7.9.5) utilized the crystalline maltodextrins only when pre-phosphorylated by GWD. Phosphorylation of crystalline maltodextrins, as catalyzed by GWD, initiated solubilization of neutral as well as phosphorylated glucans. In both the insoluble and the soluble state, mono-, di and triphosphorylated alpha-glucans were observed, with wide and overlapping ranges of degree of polymerization. Thus, the substrate specificity of the GWD is defined by the physical arrangement of alpha-glucans rather than by structural parameters, such as the distribution of branching points or degree of polymerization. Unlike GWD and PWD, recombinant beta-amylase isozyme 3 (BAM3), which has been shown to be essential for plastidial starch degradation, preferentially degraded soluble maltodextrins rather than crystallized glucans. In summary, two conclusions were reached. Firstly, carbohydrate targets of GWD are primarily defined by the molecular order of glucan helices. Secondly, GWD catalyzed phosphorylation mediates the phase transition of glucans from a highly ordered to a less ordered and hydrated state. PMID- 18419780 TI - AtRECQ2, a RecQ helicase homologue from Arabidopsis thaliana, is able to disrupt various recombinogenic DNA structures in vitro. AB - RecQ helicases play an important role in the maintenance of genomic stability in pro- and eukaryotes. This is highlighted by the human genetic diseases Werner, Bloom's and Rothmund-Thomson syndrome, caused by respective mutations in three of the five human RECQ genes. The highest numbers of RECQ homologous genes are found in plants, e.g. seven in Arabidopsis thaliana. However, only limited information is available on the functions of plant RecQ helicases, and no biochemical characterization has been performed. Here, we demonstrate that AtRECQ2 is a (d)NTP-dependent 3'-->5' DNA helicase. We further characterized its basal properties and its action on various partial DNA duplexes. Importantly, we demonstrate that AtRECQ2 is able to disrupt recombinogenic structures: by disrupting various D-loop structures, AtRECQ2 may prevent non-productive recombination events on the one hand, and may channel repair processes into non recombinogenic pathways on the other hand, thus facilitating genomic stability. We show that a synthetic partially mobile Holliday junction is processed towards splayed-arm products, possibly indicating a branch migration function for AtRECQ2. The biochemical properties defined in this work support the hypothesis that AtRECQ2 might be functionally orthologous to the helicase part of the human RecQ homologue HsWRN. PMID- 18419781 TI - The AtGenExpress hormone and chemical treatment data set: experimental design, data evaluation, model data analysis and data access. AB - We analyzed global gene expression in Arabidopsis in response to various hormones and in related experiments as part of the AtGenExpress project. The experimental agents included seven basic phytohormones (auxin, cytokinin, gibberellin, brassinosteroid, abscisic acid, jasmonate and ethylene) and their inhibitors. In addition, gene expression was investigated in hormone-related mutants and during seed germination and sulfate starvation. Hormone-inducible genes were identified from the hormone response data. The effects of each hormone and the relevance of the gene lists were verified by comparing expression profiles for the hormone treatments and related experiments using Pearson's correlation coefficient. This approach was also used to analyze the relationships among expression profiles for hormone responses and those included in the AtGenExpress stress-response data set. The expected correlations were observed, indicating that this approach is useful to monitor the hormonal status in the stress-related samples. Global interactions among hormones-inducible genes were analyzed in a pairwise fashion, and several known and novel hormone interactions were detected. Genome-wide transcriptional gene-to-gene correlations, analyzed by hierarchical cluster analysis (HCA), indicated that our data set is useful for identification of clusters of co-expressed genes, and to predict the functions of unknown genes, even if a gene's function is not directly related to the experiments included in AtGenExpress. Our data are available online from AtGenExpressJapan; the results of genome-wide HCA are available from PRIMe. The data set presented here will be a versatile resource for future hormone studies, and constitutes a reference for genome-wide gene expression in Arabidopsis. PMID- 18419782 TI - Nucleocytoplasmic-localized acyltransferases catalyze the malonylation of 7-O glycosidic (iso)flavones in Medicago truncatula. AB - (Iso)flavonoids are commonly accumulated as malonylated or acetylated glycoconjugates in legumes. Sequence analysis on EST database of the model legume Medicago truncatula enabled us to identify nine cDNA sequences encoding BAHD super-family enzymes that are distinct from the most of the characterized anthocyanin/flavonol acyltransferase genes in other species. Functional characterization revealed that three of these corresponding enzymes, MtMaT1, 2 and 3, specifically recognize malonyl CoA as an acyl donor and catalyze the malonylation of a range of isoflavone 7-O-glucosides in vitro. These malonyltransferase genes displayed distinct tissue-specific expression patterns and responded differentially to biotic and abiotic stresses. Consistent with gene expression, the level of the accumulated malonyl isoflavone glucoside was altered in the roots of M. truncatula grown under normal and drought-stressed conditions. Overexpression of the MtMaT1 gene in a previously engineered Arabidopsis line that accumulates genistein glycosides (Proc. Natl Acad. Sci. USA, 99, 2002:14578) led to a malonylated product. Confocal microscopy of the transiently expressed MtMaT1-GFP fusion revealed strong fluorescence in both the cytoplasm and nucleus of M. truncatula and tobacco leaf cells. A truncated MtMaT1 lacking the C terminal polypeptide of 110 amino acid residues that include the DFGWG motif, the single conserved sequence signature of BAHD super-family members, retained considerable catalytic efficiency, but showed an altered optimum pH preference for maximum activity. Such C-terminal polypeptide deletion or deletion of the DFGWG motif alone led to improper folding of the transiently expressed GFP fusion protein in living cells, and impaired nuclear localization of the enzyme. PMID- 18419783 TI - Ghrelin in small-for-gestational age (SGA) newborn babies: a cross-sectional study. AB - BACKGROUND: Small-for-gestational newborn babies may have long-term metabolic consequences. Among the main hormones possibly involved in foetal growth regulation are the IGFs, IGFBPs and the recently described ghrelin. OBJECTIVE: To examine the levels of desacyl-ghrelin, IGF-1, IGFBP-1 and IGFBP-3 in children Small-for-gestational age (SGA) and children adequate-for-gestational age (AGA). DESIGN: This was a cross-sectional and comparative study. SUBJECTS: We included 40 children SGA and 40 children AGA. MEASUREMENTS: Blood sample was taken a week after birth and measuring hormonal levels were done by ELISA. RESULTS: SGA babies had lower IGF-1, IGFBP-3 and leptin levels, but higher ghrelin and IGFBP-1 levels. Birth weight was associated independently with ghrelin and IGFBP-1 (negatively) and IGFBP-3 (positively). Ghrelin circulating levels were associated negative and independently with IGFBP-3 and triglycerides in the mother. CONCLUSIONS: We interpreted these findings to indicate that diminished body weight in newborns induce different adaptive signals, some of them mediated by IGF-1/IGFBP-3, ghrelin or by IGFBP-1. This regulation is congruent with the proposed role of ghrelin to adaptation to under-nutrition favouring lipid accumulation. CONCyTEG grant number 05-16-K117-028. PMID- 18419784 TI - Serum levels of adipocyte fatty acid-binding protein (AFABP) are increased in chronic haemodialysis (CD). AB - OBJECTIVE: Adipocyte fatty acid-binding protein (AFABP) was recently introduced as an adipocyte-expressed factor, serum levels of which independently correlate with the development of the metabolic syndrome and cardiovascular disease in humans. In the current study, we investigated renal elimination of this protein by comparing circulating AFABP levels in patients on chronic haemodialysis (CD) with controls. We hypothesized that if renal filtration is a significant route of elimination of AFABP, it would accumulate in CD patients. PATIENTS AND MEASUREMENTS: AFABP was determined by ELISA in control (n = 60) and CD (n = 60) patients and correlated to clinical and biochemical measures of renal function, glucose and lipid metabolism, as well as inflammation, in both groups. RESULTS: Median serum AFABP levels were more than 10-fold higher in CD patients (510.9 +/- 294.7 microg/l) as compared to controls (44.3 +/- 35.2 microg/l). Furthermore, CD independently predicted AFABP concentrations in multiple regression analysis. In addition, body mass index and free fatty acids were independently associated with circulating AFABP. CONCLUSIONS: Renal filtration appears as an important route of elimination of AFABP. Future studies should elucidate whether this adipocyte expressed factor contributes to the increased risk of cardiovascular disease found in end-stage renal disease. PMID- 18419785 TI - Incidence of radiation thyroiditis and thyroid remnant ablation success rates following 1110 MBq (30 mCi) and 3700 MBq (100 mCi) post-surgical 131I ablation therapy for differentiated thyroid carcinoma. AB - OBJECTIVE: To evaluate the relationships between thyroid remnant (131)I uptake, radiation thyroiditis and remnant ablation success rate between lower (1110 MBq) and higher (3700 MBq) initial ablative (131)I dose for post-surgical ablation therapy for differentiated thyroid cancer. DESIGN: Patients having post-surgical administration of 1110 MBq (68 patients) or 3700 MBq (115 patients) (131)I were retrospectively reviewed. Thyroid remnant (131)I uptake on a 48 h post administration scan was correlated with neck symptoms experienced. Patients were classified as having insignificant, mild or severe thyroiditis based on symptoms. Absent thyroid bed (131)I uptake on a follow-up 74 MBq (131)I study was considered successful ablation. RESULTS: 183 patients were included. Median (131)I remnant uptake was 37 MBq. 21% (39/183) of patients developed thyroiditis. Incidence and severity of thyroiditis increased with increasing remnant (131)I activity (P 73 MBq. For patients treated with 1110 MBq and 3700 MBq, incidence of thyroiditis was 12% and 27% (P = 0.02) and remnant ablation success rate was 76% and 84% (P = NS), respectively. Occurrence of thyroiditis did not correlate with successful ablation. CONCLUSIONS: Incidence and severity of radiation thyroiditis following (131)I remnant ablation therapy is directly related to thyroid remnant (131)I uptake. As 1110 MBq (131)I is associated with a significantly lower frequency of thyroiditis but similar remnant ablation rate to 3700 MBq, it warrants consideration for thyroid remnant ablation particularly in patients with low risk disease. PMID- 18419786 TI - Acylated ghrelin decreases during acute exercise in the lean and obese state. PMID- 18419787 TI - Clinical manifestations of familial paraganglioma and phaeochromocytomas in succinate dehydrogenase B (SDH-B) gene mutation carriers. AB - OBJECTIVE: Phaeochromocytomas and paragangliomas are familial in up to 25% of cases and can result from succinate dehydrogenase (SDH) gene mutations. The aim of this study was to describe the clinical manifestations of subjects with SDH-B gene mutations. DESIGN: Retrospective case-series. PATIENTS: Thirty-two subjects with SDH-B gene mutations followed up between 1975 and 2007. Mean follow-up of 5.8 years (SD 7.4, range 0-31). Patients seen at St Bartholomew's Hospital, London and other UK centres. MEASUREMENTS: Features of clinical presentation, genetic mutations, tumour location, catecholamine secretion, clinical course and management. RESULTS: Sixteen of 32 subjects (50%) were affected by disease. Two previously undescribed mutations in the SDH-B gene were noted. A family history of disease was apparent in only 18% of index subjects. Mean age at diagnosis was 34 years (SD 15.4, range 10-62). 50% of affected subjects had disease by the age of 26 years. 69% (11 of 16) were hypertensive and 80% (12 of 15) had elevated secretions of catecholamines/metabolites. 24% (6 of 25) of tumours were located in the adrenal and 76% (19 of 25) were extra-adrenal. 19% (3 of 16) had multifocal disease. Metastatic paragangliomas developed in 31% (5 of 16). One subject developed a metastatic type II papillary renal cell carcinoma. The cohort malignancy rate was 19% (6 of 32). Macrovascular disease was noted in two subjects without hypertension. CONCLUSION: SDH-B mutation carriers develop disease early and predominantly in extra-adrenal locations. Disease penetrance is incomplete. Metastatic disease is prominent but levels are less than previously reported. Clinical manifestations may include papillary renal cell carcinoma and macrovascular disease. PMID- 18419788 TI - Detection of somatic beta-catenin mutations in primary pigmented nodular adrenocortical disease (PPNAD). AB - BACKGROUND: Primary pigmented nodular adrenocortical disease (PPNAD) leads to Cushing syndrome (CS) and is often associated with Carney complex (CNC). Genetic alterations of the type 1-alpha regulatory subunit of cAMP-dependent protein kinase A (PRKAR1A) and phosphodiesterase 11A4 (PDE11A) genes have been found in PPNAD. Recent studies have demonstrated that beta-catenin mutations are frequent in adrenocortical adenomas and carcinomas and that the Wnt-signalling pathway is involved in PPNAD tumorigenesis. We hypothesized that adrenocortical adenomas that form in the context of PPNAD may harbour beta-catenin mutations. METHODS: We studied 18 patients with CS secondary to PPNAD who were screened for germline PRKAR1A and PDE11A mutations. Tumor DNA was extracted from pigmented adrenocortical adenoma and nodular adrenal hyperplasia. Mutation analysis of exons 3 and 5 of beta-catenin was performed using polymerase chain reaction and direct sequencing. Sections from formalin-fixed, paraffin-embedded tumour samples were studied by immunohistochemistry with an antibody against beta-catenin. RESULTS: Nine patients were carrying germline PRKAR1A mutations and one patient had a PDE11A mutation. We found somatic beta-catenin mutations in 2 of 18 patients (11%). In both cases, the mutations occurred in relatively large adenomas that had formed in the background of PPNAD. Tumor DNA analysis revealed a heterozygous ACC-to-GCC missense mutation in codon 41 (T41A) and a TCT-to-CCT missense mutation in codon 45 (S45P) of exon 3 of the beta-catenin gene that was confirmed at the cDNA level. There were no alterations in the DNA of PPNAD adjacent tissues and lymphocytes from the patients, indicating somatic events. Immunohistochemistry showed nuclear accumulation of beta-catenin in more than 90% of cells in adenomatous tissue whereas no nuclear immunoreactivity was detected in adjacent PPNAD nodular cells. Nuclear translocation of beta-catenin protein in the PPNAD adenoma suggests activation of the Wnt-beta-catenin pathway in PPNAD. CONCLUSIONS: We report, for the first time, beta-catenin mutations in adenomas associated with PPNAD, further implicating Wnt-beta-catenin signalling in tumorigenesis linked to bilateral adrenal hyperplasias. PMID- 18419789 TI - Relationship between calcium absorption and plasma dehydroepiandrosterone sulphate (DHEAS) in healthy males. AB - CONTEXT: Impaired gut sensitivity to 1,25-dihydroxyvitamin D (1,25(OH)(2)D), leading to reduced intestinal calcium absorption, has been reported in older men and women. While this phenomenon in postmenopausal women has been attributed to oestrogen deficiency, it is unclear whether the same observation in older men correlates with the age-related decline in androgen concentrations. OBJECTIVE: To examine the relationship between androgens and intestinal calcium absorption in older men. DESIGN: Cross-sectional study on 55 healthy male volunteers, divided into younger (n = 27) and older (n = 28) groups separated according to the median age of 59 years. MAIN OUTCOME MEASURES: Calcium absorption, total and free (calculated) testosterone, dehydroepiandrosterone sulphate (DHEAS), SHBG, and 1,25(OH)(2)D, among others, were measured. RESULTS: Calcium absorption, free testosterone and DHEAS, but not 1,25(OH)(2)D, declined significantly with age. After adjusting for age and body mass index, stepwise regression showed that 1,25(OH)(2)D and serum albumin were the only significant determinants of calcium absorption in younger men, while the sole determinant in older men was DHEAS, not testosterone. Residual deviations from the regression of calcium absorption on 1,25(OH)(2)D, reflecting the efficiency of 1,25(OH)(2)D-induced calcium absorption, was positively correlated with DHEAS (r = 0.27, P = 0.027). CONCLUSIONS: DHEAS is an independent determinant of calcium absorption in older men, although its manner of influence is, as yet, undefined. The age-related decline of DHEAS may, partly, account for the observed 'intestinal resistance to 1,25(OH)(2)D' in older men. PMID- 18419790 TI - Secondary hyperparathyroidism in primary osteoporosis and osteopenia: optimizing calcium and vitamin D intakes to levels recommended by expert panels may not be sufficient for correction. AB - OBJECTIVE: To compare biochemical variables, renal function and calcium and vitamin D intakes in euparathyroid and hyperparathyroid patients with primary osteoporosis and osteopenia and describe the measures necessary to normalize serum PTH in the patients with secondary hyperparathyroidism. DESIGN AND PATIENTS: We reviewed the charts of normocalcemic patients with primary osteoporosis and osteopenia first seen during the years 1991-2003 and identified 75 with elevated serum PTH levels at baseline. These patients were compared to all the 143 euparathyroid patients first seen in 1998 and 1999. Patients were restudied after 1 year and we attempted to follow patients with secondary hyperparathyroidism until PTH levels became normal. MEASUREMENTS: At baseline serum PTH, ionized calcium, inorganic phosphate, alkaline phosphatase, creatinine, a complete blood count and serum 25 hydroxy vitamin D were measured in the early morning fasting state. These tests were repeated at follow up. RESULTS: In one-third of the hyperparathyroid patients, the standard baseline treatment failed to correct the secondary hyperparathyroidism necessitating extraordinary measures including unusually large doses of vitamin D (i.e. 50 000 IU vitamin D(2) twice weekly) or the substitution of calcium citrate for calcium carbonate as a calcium supplement. CONCLUSION: Large doses of vitamin D are frequently necessary to suppress secondary hyperparathyroidism in patients with primary osteoporosis and osteopenia. This suggests that vitamin D metabolism may be altered in some of these patients. PMID- 18419791 TI - Nogo-A and Nogo-66 receptor in amyotrophic lateral sclerosis. AB - Nogo/reticulon (RTN)-4 has been strongly implicated as a disease marker for the motor neuron disease amyotrophic lateral sclerosis (ALS). Nogo isoforms, including Nogo-A, are ectopically expressed in the skeletal muscle of ALS mouse models and patients and their levels correlate with the disease severity. The notion of a direct involvement of Nogo-A in ALS aetiology is supported by the findings that Nogo-A deletion in mice reduces muscle denervation and prolongs survival, whereas overexpression of Nogo-A destabilizes motor nerve terminals and promotes denervation. Another intriguing, and somewhat paradoxical, recent finding revealed that binding of the Nogo-66 receptor (NgR) by either agonistic or antagonistic Nogo-66-derived peptides protects against p75 neurotrophin receptor (p75(NTR))-dependent motor neuron death. Ligand binding by NgR could result in subsequent engagement of p75(NTR), and this association could preclude pro-apoptotic signalling by the latter. Understanding the intricate interplay among Nogo isoforms, NgR and p75(NTR) in ALS disease progression may provide important, therapeutically exploitable information. PMID- 18419793 TI - Immune-based disorders: the challenges for translational immunology. PMID- 18419792 TI - The alternative complement pathway revisited. AB - Alternative pathway amplification plays a major role for the final effect of initial specific activation of the classical and lectin complement pathways, but the quantitative role of the amplification is insufficiently investigated. In experimental models of human diseases in which a direct activation of alternative pathway has been assumed, this interpretation needs revision placing a greater role on alternative amplification. We recently documented that the alternative amplification contributed to 80-90% of C5 activation when the initial activation was highly specific for the classical pathway. The recent identification of properdin as a recognition factor directly initiating alternative pathway activation, like C1q in the classical and mannose-binding lectin in the lectin pathway initiates a renewed interest in the reaction mechanisms of complement. Complement and Toll-like receptors, including the CD14 molecule, are two main upstream recognition systems of innate immunity, contributing to the inflammatory reaction in a number of conditions including ischemia-reperfusion injury and sepsis. These systems act as "double-edged swords", being protective against microbial invasion, but harmful to the host when activated improperly or uncontrolled. Combined inhibition of complement and Toll-like receptors/CD14 should be explored as a treatment regimen to reduce the overwhelming damaging inflammatory response during sepsis. The alternative pathway should be particularly considered in this regard, due to its uncontrolled amplification in sepsis. The alternative pathway should be regarded as a dual system, namely a recognition pathway principally similar to the classical and lectin pathways, and an amplification mechanism, well known, but quantitatively probably more important than generally recognized. PMID- 18419794 TI - Osteopontin and oral carcinogenesis. PMID- 18419796 TI - Apolipoprotein D is involved in the mechanisms regulating protection from oxidative stress. AB - Many nervous system pathologies are associated with increased levels of apolipoprotein D (ApoD), a lipocalin also expressed during normal development and aging. An ApoD homologous gene in Drosophila, Glial Lazarillo, regulates resistance to stress, and neurodegeneration in the aging brain. Here we study for the first time the protective potential of ApoD in a vertebrate model organism. Loss of mouse ApoD function increases the sensitivity to oxidative stress and the levels of brain lipid peroxidation, and impairs locomotor and learning abilities. Human ApoD overexpression in the mouse brain produces opposite effects, increasing survival and preventing the raise of brain lipid peroxides after oxidant treatment. These observations, together with its transcriptional up regulation in the brain upon oxidative insult, identify ApoD as an acute response protein with a protective and therefore beneficial function mediated by the control of peroxidated lipids. PMID- 18419795 TI - Mouse models with human immunity and their application in biomedical research. AB - Biomedical research in human beings is largely restricted to in vitro studies that lack complexity of a living organism. To overcome this limitation, humanized mouse models are developed based on immunodeficient characteristics of severe combined immunodeficiency (SCID) or recombination activating gene (Rag)(null) mice, which can accept xenografts. Peripheral constitution of human immunity in SCID or Rag(null) mice has been achieved by transplantation of mature human immune cells, foetal human thymus, bone marrow, liver tissues, lymph nodes or a combination of these, although efficiency needs to be improved. These mouse models with constituted human immunity (defined as humanized mice in the present text) have been widely used to investigate the basic principles of human immunobiology as well as complex pathomechanisms and potential therapies of human diseases. Here, elements of an ideal humanized mouse model are highlighted including genetic and non-genetic modification of recipient mice, transplantation strategies and proposals to improve engraftments. The applications of the humanized mice to study the development and response of human immune cells, human autoimmune diseases, virus infections, transplantation biology and tumour biology are reviewed as well. PMID- 18419797 TI - Age-related increase of superoxide generation in the brains of mammals and birds. AB - Oxidative stress, an imbalance between endogenous levels of oxygen radicals and antioxidative defense, increases with aging. However, it is not clear which of these two factors is the more critical. To clarify the production of oxygen radicals increases with age, we examined oxygen radical-dependent chemiluminescent signals in ex vivo brain slices using a novel photonic imaging method. The chemiluminescent intensity was significantly decreased by the membrane permeable superoxide dismutase (SOD)/catalase mimic, but not by Cu,Zn SOD. Inhibitors for complex I, III, and IV of the mitochondrial electron transport chain transiently enhanced the chemiluminescent signal. The superoxide dependent chemiluminescent intensity in senescence accelerated mouse (SAM) brain tissues increases with age. Moreover, the slope of the age-dependent increase was steeper in SAMP10, a strain characterized by a short lifespan and atrophy in the frontal cerebral cortex, than the senescence-resistant strain SAMR1, which has a longer lifespan. An increase in chemiluminescence with age was also observed in C57/BL6 mice, Wistar rats, and pigeons, although levels of chemiluminescence were lower in the pigeons than murines. The rate of age-related increases of superoxide-dependent chemiluminescence was inversely related to the maximum lifespan of the animals. The activity of superoxide dismutase was unchanged during the aging process in the brain. This suggested that superoxide production itself may increase with age. We speculated that reactive oxygen may be a signal to determine the aging process. PMID- 18419798 TI - Maternal chronic viral infections transmitted to infants: from mechanism to prevention and care. Abstracts of the Fourth Dominique Dormont International Conference. Paris, France. December 13-15, 2007. PMID- 18419799 TI - Tone burst-evoked otoacoustic emissions in neonates: normative data. AB - BACKGROUND: Tone-burst otoacoustic emissions (TBOAEs) have not been routinely studied in pediatric populations, although tone burst stimuli have greater frequency specificity compared with click sound stimuli. The present study aimed (1) to determine an appropriate stimulus level for neonatal TBOAE measurements when the stimulus center frequency was 1 kHz, (2) to explore the characteristics of 1 kHz TBOAEs in a neonatal population. METHODS: A total of 395 normal neonates (745 ears) were recruited. The study consisted of two parts, reflecting the two study aims. Part I included 40 normal neonatal ears, and TBOAE measurement was performed at five stimulus levels in the range 60-80 dB peSPL, with 5 dB incremental steps. Part II investigated the characteristics of the 1 kHz TBOAE response in a large group of 705 neonatal ears, and provided clinical reference criteria based on these characteristics. RESULTS: The study provided a series of reference parameters for 1 kHz TBOAE measurement in neonates. Based on the results, a suggested stimulus level and reference criteria for 1 kHz TBOAE measures with neonates were established. In addition, time-frequency analysis of the data gave new insight into the energy distribution of the neonatal TBOAE response. CONCLUSION: TBOAE measures may be a useful method for investigating cochlear function at specific frequency ranges in neonates. However, further studies of both TBOAE time-frequency analysis and measurements in newborns are needed. PMID- 18419800 TI - Genotyping of Chlamydophila psittaci using a new DNA microarray assay based on sequence analysis of ompA genes. AB - BACKGROUND: The currently used genotyping system for the avian zoonotic pathogen Chlamydophila (C.) psittaci has evolved from serology and is based on ompA sequence variations. It includes seven avian and two non-avian genotypes. Restriction enzyme cleavage of the amplified ompA gene and, less frequently, ompA sequencing are being used for examination, but, beside methodological limitations, an increasing number of recently tested strains could not be assigned to any established genotype. RESULTS: Comprehensive analysis of all available ompA gene sequences has revealed a remarkable genetic diversity within the species C. psittaci, which is only partially covered by the present genotyping scheme. We suggest adjustments and extensions to the present scheme, which include the introduction of subgroups to the more heterogeneous genotypes A, E/B and D, as well as six provisional genotypes representing so far untypable strains. The findings of sequence analysis have been incorporated in the design of a new DNA microarray. The ArrayTubetrade mark microarray-based ompA genotyping assay has been shown to discriminate among established genotypes and identify so far untyped strains. Its high specificity, which allows detection of single nucleotide polymorphisms, is due to the parallel approach consisting in the use of 35 hybridization probes derived from variable domains 2 and 4 of the ompA gene. CONCLUSION: The traditional genotyping system does not adequately reflect the extent of intra-species heterogeneity in ompA sequences of C. psittaci. The newly developed DNA microarray-based assay represents a promising diagnostic tool for tracing epidemiological chains, exploring the dissemination of genotypes and identifying non-typical representatives of C. psittaci. PMID- 18419801 TI - Predictive gene lists for breast cancer prognosis: a topographic visualisation study. AB - BACKGROUND: The controversy surrounding the non-uniqueness of predictive gene lists (PGL) of small selected subsets of genes from very large potential candidates as available in DNA microarray experiments is now widely acknowledged 1. Many of these studies have focused on constructing discriminative semi parametric models and as such are also subject to the issue of random correlations of sparse model selection in high dimensional spaces. In this work we outline a different approach based around an unsupervised patient-specific nonlinear topographic projection in predictive gene lists. METHODS: We construct nonlinear topographic projection maps based on inter-patient gene-list relative dissimilarities. The Neuroscale, the Stochastic Neighbor Embedding(SNE) and the Locally Linear Embedding(LLE) techniques have been used to construct two dimensional projective visualisation plots of 70 dimensional PGLs per patient, classifiers are also constructed to identify the prognosis indicator of each patient using the resulting projections from those visualisation techniques and investigate whether a-posteriori two prognosis groups are separable on the evidence of the gene lists.A literature-proposed predictive gene list for breast cancer is benchmarked against a separate gene list using the above methods. Generalisation ability is investigated by using the mapping capability of Neuroscale to visualise the follow-up study, but based on the projections derived from the original dataset. RESULTS: The results indicate that small subsets of patient-specific PGLs have insufficient prognostic dissimilarity to permit a distinction between two prognosis patients. Uncertainty and diversity across multiple gene expressions prevents unambiguous or even confident patient grouping. Comparative projections across different PGLs provide similar results. CONCLUSION: The random correlation effect to an arbitrary outcome induced by small subset selection from very high dimensional interrelated gene expression profiles leads to an outcome with associated uncertainty. This continuum and uncertainty precludes any attempts at constructing discriminative classifiers.However a patient's gene expression profile could possibly be used in treatment planning, based on knowledge of other patients' responses.We conclude that many of the patients involved in such medical studies are intrinsically unclassifiable on the basis of provided PGL evidence. This additional category of 'unclassifiable' should be accommodated within medical decision support systems if serious errors and unnecessary adjuvant therapy are to be avoided. PMID- 18419802 TI - Vitamin D receptor gene polymorphisms and haplotypes and postmenopausal breast cancer risk. AB - INTRODUCTION: Vitamin D receptor (VDR) genotypes may influence breast cancer risk by altering potential anticarcinogenic effects of vitamin D, but epidemiological studies have been inconsistent. Effect modification by serum 25-hydroxyvitamin D (25 [OH]D), the biomarker for vitamin D status in humans, has rarely been examined. METHODS: We assessed the effects of two frequently analyzed polymorphisms (FokI and TaqI) and two potentially functional variants (VDR-5132 and Cdx2) in the VDR gene, which thus far have not been analyzed with respect to breast cancer risk, on postmenopausal breast cancer risk in a population-based, case-control study including 1,408 patients (cases) and 2,612 control individuals (controls) matched for year of birth. Odds ratios (ORs) for breast cancer adjusted for potential confounders were calculated for genotypes and estimated haplotypes. RESULTS: No differences in serum 25(OD)D concentrations by VDR genotype were observed. None of the analyzed polymorphisms was associated with overall risk for postmenopausal breast cancer. However, the TaqI polymorphism was associated with a significantly increased risk for oestrogen receptor positive tumours (OR = 1.18, 95% confidence interval [CI] = 1.00 to 1.38, comparing t allele carriers with noncarriers) but not for oestrogen receptor negative tumours (OR = 0.88, 95% CI = 0.69 to 1.13; P for interaction = 0.04). Haplotype analysis revealed the haplotype FtCA (FokI F, TaqI t, VDR-5132 C, Cdx2 A), which contains the TaqI t allele, to be associated with a significantly greater breast cancer risk as compared with the most frequent haplotype FTCG (OR = 1.43, 95% CI = 1.00 to 2.05). No significant interaction between VDR genotypes or haplotypes and 25(OH)D was observed. CONCLUSION: Our results support potential effects of VDR polymorphisms on postmenopausal breast cancer risk and possible differential effects of receptor status of the tumour. However, further studies focusing on the influence of polymorphisms and haplotypes on VDR functionality, activity and concentration are needed. PMID- 18419803 TI - Tumor necrosis factor alpha drugs in rheumatoid arthritis: systematic review and metaanalysis of efficacy and safety. AB - BACKGROUND: To analyse available evidence on the efficacy and safety of anti TNFalpha drugs (infliximab, etanercept and adalimumab) for treating rheumatoid arthritis (RA). METHODS: We searched systematically for randomised controlled clinical trials on treatment of RA with anti-TNFalpha drugs, followed by a systematic review with metaanalysis. Trials were searched from MEDLINE, EMBASE and Cochrane Library databases. The American College of Rheumatology (ACR) efficacy response criteria were used. Safety parameters provided by the trials were also assessed. Positive and undesired effects were estimated using combined relative risks (RR), number needed to treat (NNT) and number needed to harm (NNH). Heterogeneity was evaluated by Cochrane's Q and I2 statistics. RESULTS: Thirteen trials (7087 patients) met the inclusion criteria. The combined RR to achieve a therapeutic response to treatment with recommended doses of any anti TNFalpha drug was 1.81 (95% CI 1.43-2.29) with a NNT of 5 (5-6) for ACR20. NNT for ACR50 [5 (5-6)] and ACR70 [7 (7-9)] were similar. Overall therapeutic effects were also similar regardless of the specific anti-TNFalpha drug used and when higher than recommended doses were administered. However, lower than recommended doses elicited low ACR70 responses (NNT 15). Comparison of anti-TNFalpha drugs plus methotrexate (MTX) with MTX alone in patients with insufficient prior responses to MTX showed NNT values of 3 for ACR20, 4 for ACR50 and 8 for ACR70. Comparison of anti-TNFalpha drugs with placebo showed a similar pattern. Comparisons of anti-TNFalpha drugs plus MTX with MTX alone in patients with no previous resistance to MTX showed somewhat lower effects. Etanercept and adalimumab administered as monotherapy showed effects similar to those of MTX. Side effects were more common among patients receiving anti-TNFalpha drugs than controls (overall combined NNH 27). Patients receiving infliximab were more likely to drop out because of side effects (NNH 24) and to suffer severe side effects (NNH 31), infections (NNH 10) and infusion reactions (NNH 9). Patients receiving adalimumab were also more likely to drop out because of side effects (NNH 47) and to suffer injection site reactions (NNH 22). Patients receiving etanercept were less likely to drop out because of side effects (NNH for control versus etanercept 26) but more likely to experience injection site reactions (NNH 5). CONCLUSION: Anti-TNFalpha drugs are effective in RA patients, with apparently similar results irrespective of the drug administered. Doses other than those recommended are also beneficial. The main factor influencing therapeutic efficacy is the prior response to DMARD treatment. The effect of treatment with etanercept or adalimumab does not differ from that obtained with MTX. The published safety profile for etanercept is superior but the fact that no patients are treated with higher than recommended doses requires explanation. PMID- 18419804 TI - Training tomorrow's doctors in diabetes: self-reported confidence levels, practice and perceived training needs of post-graduate trainee doctors in the UK. A multi-centre survey. AB - OBJECTIVE: To assess the confidence, practices and perceived training needs in diabetes care of post-graduate trainee doctors in the UK. METHODS: An anonymised postal questionnaire using a validated 'Confidence Rating' (CR) scale was applied to aspects of diabetes care and administered to junior doctors from three UK hospitals. The frequency of aspects of day-to-day practice was assessed using a five-point scale with narrative description in combination with numeric values. Respondents had a choice of 'always' (100%), 'almost always' (80-99%), 'often' (50-79%), 'not very often' (20-49%) and 'rarely' (less than 20%). Yes/No questions were used to assess perception of further training requirements. Additional 'free-text' comments were also sought. RESULTS: 82 doctors completed the survey. The mean number of years since medical qualification was 3 years and 4 months, (range: 4 months to 14 years and 1 month). Only 11 of the respondents had undergone specific diabetes training since qualification.4(5%) reported 'not confident' (CR1), 30 (37%) 'satisfactory but lacked confidence' (CR2), 25 (30%) felt 'confident in some cases' (CR3) and 23 (28%) doctors felt fully confident (CR4) in diagnosing diabetes. 12 (15%) doctors would always, 24 (29%) almost always, 20 (24%) often, 22 (27%) not very often and 4 (5%) rarely take the initiative to optimise gcaemic control. 5 (6%) reported training in diagnosis of diabetes was adequate while 59 (72%) would welcome more training. Reported confidence was better in managing diabetes emergencies, with 4 (5%) not confident in managing hypoglycaemia, 10 (12%) lacking confidence, 22 (27%) confident in some cases and 45 (55%) fully confident in almost all cases. Managing diabetic ketoacidosis, 5 (6%) doctors did not feel confident, 16 (20%) lacked confidence, 20 (24%) confident in some cases, and 40 (50%) felt fully confident in almost all cases. CONCLUSION: There is a lack of confidence in managing aspects of diabetes care, including the management of diabetes emergencies, amongst postgraduate trainee doctors with a perceived need for more training. This may have considerable significance and further research is required to identify the causes of deficiencies identified in this study. PMID- 18419805 TI - The small molecule Mek1/2 inhibitor U0126 disrupts the chordamesoderm to notochord transition in zebrafish. AB - BACKGROUND: Key molecules involved in notochord differentiation and function have been identified through genetic analysis in zebrafish and mice, but MEK1 and 2 have so far not been implicated in this process due to early lethality (Mek1-/-) and functional redundancy (Mek2-/-) in the knockout animals. RESULTS: Here, we reveal a potential role for Mek1/2 during notochord development by using the small molecule Mek1/2 inhibitor U0126 which blocks phosphorylation of the Mek1/2 target gene Erk1/2 in vivo. Applying the inhibitor from early gastrulation until the 18-somite stage produces a specific and consistent phenotype with lack of dark pigmentation, shorter tail and an abnormal, undulated notochord. Using morphological analysis, in situ hybridization, immunhistochemistry, TUNEL staining and electron microscopy, we demonstrate that in treated embryos the chordamesoderm to notochord transition is disrupted and identify disorganization in the medial layer of the perinotochordal basement mebrane as the probable cause of the undulations and bulges in the notochord. We also examined and excluded FGF as the upstream signal during this process. CONCLUSION: Using the small chemical U0126, we have established a novel link between MAPK-signaling and notochord differentiation. Our phenotypic analysis suggests a potential connection between the MAPK-pathway, the COPI-mediated intracellular transport and/or the copper dependent posttranslational regulatory processes during notochord differentiation. PMID- 18419806 TI - Erect wing regulates synaptic growth in Drosophila by integration of multiple signaling pathways. AB - BACKGROUND: Formation of synaptic connections is a dynamic and highly regulated process. Little is known about the gene networks that regulate synaptic growth and how they balance stimulatory and restrictive signals. RESULTS: Here we show that the neuronally expressed transcription factor gene erect wing (ewg) is a major target of the RNA binding protein ELAV and that EWG restricts synaptic growth at neuromuscular junctions. Using a functional genomics approach we demonstrate that EWG acts primarily through increasing mRNA levels of genes involved in transcriptional and post-transcriptional regulation of gene expression, while genes at the end of the regulatory expression hierarchy (effector genes) represent only a minor portion, indicating an extensive regulatory network. Among EWG-regulated genes are components of Wingless and Notch signaling pathways. In a clonal analysis we demonstrate that EWG genetically interacts with Wingless and Notch, and also with TGF-beta and AP-1 pathways in the regulation of synaptic growth. CONCLUSION: Our results show that EWG restricts synaptic growth by integrating multiple cellular signaling pathways into an extensive regulatory gene expression network. PMID- 18419807 TI - The complete genome, comparative and functional analysis of Stenotrophomonas maltophilia reveals an organism heavily shielded by drug resistance determinants. AB - BACKGROUND: Stenotrophomonas maltophilia is a nosocomial opportunistic pathogen of the Xanthomonadaceae. The organism has been isolated from both clinical and soil environments in addition to the sputum of cystic fibrosis patients and the immunocompromised. Whilst relatively distant phylogenetically, the closest sequenced relatives of S. maltophilia are the plant pathogenic xanthomonads. RESULTS: The genome of the bacteremia-associated isolate S. maltophilia K279a is 4,851,126 bp and of high G+C content. The sequence reveals an organism with a remarkable capacity for drug and heavy metal resistance. In addition to a number of genes conferring resistance to antimicrobial drugs of different classes via alternative mechanisms, nine resistance-nodulation-division (RND)-type putative antimicrobial efflux systems are present. Functional genomic analysis confirms a role in drug resistance for several of the novel RND efflux pumps. S. maltophilia possesses potentially mobile regions of DNA and encodes a number of pili and fimbriae likely to be involved in adhesion and biofilm formation that may also contribute to increased antimicrobial drug resistance. CONCLUSION: The panoply of antimicrobial drug resistance genes and mobile genetic elements found suggests that the organism can act as a reservoir of antimicrobial drug resistance determinants in a clinical environment, which is an issue of considerable concern. PMID- 18419808 TI - Barriers to access prevention of mother-to-child transmission for HIV positive women in a well-resourced setting in Vietnam. AB - BACKGROUND: According to Vietnamese policy, HIV-infected women should have access at least to HIV testing and Nevirapine prophylaxis, or where available, to adequate counselling, HIV infection staging, ARV prophylaxis, and infant formula. Many studies in high HIV prevalence settings have reported low coverage of PMTCT services, but there have been few reports from low HIV prevalence settings, such as Asian countries. We investigated the access of HIV-infected pregnant women to PMTCT services in the well-resourced setting of the capital city, Hanoi. METHODS: Fifty-two HIV positive women enrolled in a self-help group in Hanoi were consulted, through in-depth interviews and bi-weekly meetings, about their experiences in accessing PMTCT services. RESULTS: Only 44% and 20% of the women had received minimal and comprehensive PMTCT services, respectively. Nine women did not receive any services. Twenty-two women received no counselling. The women reported being limited by lack of knowledge and information due to poor counselling, gaps in PMTCT services, and fear of stigma and discrimination. HIV testing was done too late for optimal interventions and poor quality of care by health staff was frequently mentioned. CONCLUSION: In a setting where PMTCT is available, HIV-infected women and children did not receive adequate care because of barriers to accessing those services. The results suggest key improvements would be improving quality of counselling and making PMTCT guidelines available to health services. Women should receive early HIV testing with adequate counselling, safe care and prophylaxis in a positive atmosphere towards HIV infected women. PMID- 18419809 TI - Foucault's "fearless speech" and the transformation and mentoring of medical students. AB - In his six 1983 lectures published under the title, Fearless Speech (2001), Michel Foucault developed the theme of free speech and its relation to frankness, truth-telling, criticism, and duty. Derived from the ancient Greek word parrhesia, Foucault's analysis of free speech is relevant to the mentoring of medical students. This is especially true given the educational and social need to transform future physicians into able citizens who practice a fearless freedom of expression on behalf of their patients, the public, the medical profession, and themselves in the public and political arena. In this paper, we argue that Foucault's understanding of free speech, or parrhesia, should be read as an ethical response to the American Medical Association's recent educational effort, Initiative to Transform Medical Education (ITME): Recommendations for change in the system of medical education (2007). In this document, the American Medical Association identifies gaps in medical education, emphasizing the need to enhance health system safety and quality, to improve education in training institutions, and to address the inadequacy of physician preparedness in new content areas. These gaps, and their relationship to the ITME goal of promoting excellence in patient care by implementing reform in the US system of medical education, call for a serious consideration and use of Foucault's parrhesia in the way that medical students are trained and mentored. PMID- 18419810 TI - Epidemiological analysis of injury in Shandong Province, China. AB - BACKGROUND: Injury is an emerging public health problem with social development and modernization in developing countries. To describe the prevalence and burden of injury and provide elaborate information for policy development, we conducted a community-based household survey in the Shandong Province of China. METHODS: The survey was conducted in 2004. Participants were selected by a multi-stage random sampling method. Information on injuries occurring in 2003 was collected in four cities and six rural counties in Shandong Province, China. RESULTS: The estimated incidence rate of injury in Shandong Province was 67.7 per 1,000. Injury incidence was higher in rural areas (84.3 per 1,000) than in urban areas (42.9 per 1,000), and was higher among males (81.1 per 1,000) than females (54.1 per 1,000). The average years of potential life lost is 37.7 years for each fatal injury. All injuries together caused 6,080,407 RMB yuan of direct and indirect economic loss, with traffic injuries accounting for 44.8% of the total economic loss. CONCLUSION: Injury incidence was higher among males than females, and in rural areas than in urban areas. Youngsters suffered the highest incidence of injury. Injury also caused large losses in terms of both economics and life, with traffic injuries contributing the most to this loss. Strategies for prevention of injury should be developed. PMID- 18419811 TI - Berry and phenology-related traits in grapevine (Vitis vinifera L.): from quantitative trait loci to underlying genes. AB - BACKGROUND: The timing of grape ripening initiation, length of maturation period, berry size and seed content are target traits in viticulture. The availability of early and late ripening varieties is desirable for staggering harvest along growing season, expanding production towards periods when the fruit gets a higher value in the market and ensuring an optimal plant adaptation to climatic and geographic conditions. Berry size determines grape productivity; seedlessness is especially demanded in the table grape market and is negatively correlated to fruit size. These traits result from complex developmental processes modified by genetic, physiological and environmental factors. In order to elucidate their genetic determinism we carried out a quantitative analysis in a 163 individuals F1 segregating progeny obtained by crossing two table grape cultivars. RESULTS: Molecular linkage maps covering most of the genome (2n = 38 for Vitis vinifera) were generated for each parent. Eighteen pairs of homologous groups were integrated into a consensus map spanning over 1426 cM with 341 markers (mainly microsatellite, AFLP and EST-derived markers) and an average map distance between loci of 4.2 cM. Segregating traits were evaluated in three growing seasons by recording flowering, veraison and ripening dates and by measuring berry size, seed number and weight. QTL (Quantitative Trait Loci) analysis was carried out based on single marker and interval mapping methods. QTLs were identified for all but one of the studied traits, a number of them steadily over more than one year. Clusters of QTLs for different characters were detected, suggesting linkage or pleiotropic effects of loci, as well as regions affecting specific traits. The most interesting QTLs were investigated at the gene level through a bioinformatic analysis of the underlying Pinot noir genomic sequence. CONCLUSION: Our results revealed novel insights into the genetic control of relevant grapevine features. They provide a basis for performing marker-assisted selection and testing the role of specific genes in trait variation. PMID- 18419812 TI - Long-term outcome of infective endocarditis: a study on patients surviving over one year after the initial episode treated in a Finnish teaching hospital during 25 years. AB - BACKGROUND: Only a few previous studies have focused on the long-term prognosis of the patients with infective endocarditis (IE). Our purpose was to delineate factors potentially associated with the long-term outcome of IE, recurrences of IE and requirement for late valve surgery. METHODS: A total of 326 episodes of IE in 303 patients were treated during 1980-2004 in the Turku University Hospital. We evaluated the long-term outcome and requirement for late valve surgery for 243 of these episodes in 226 patients who survived longer than 1 year after the initial admission. Factors associated with recurrences were analysed both for the 1-year survivors and for all 303 patients. RESULTS: The mean (SD) follow-up time for the 1-year survivors was 11.5 (7.3) years (range 25 days to 25.5 years). The overall survival was 95%, 82%, 66%, 51% and 45% at 2, 5, 10, 15 and 20 years. In age and sex adjusted multivariate analyses, significant predictors for long-term overall mortality were heart failure within 3 months of admission (HR 1.97, 95% CI 1.27 to 3.06; p = 0.003) and collagen disease (HR 2.54, 95% CI 1.25 to 5.19; p = 0.010) or alcohol abuse (HR 2.39, 95% CI 1.30 to 4.40; p = 0.005) as underlying conditions, while early surgery was significantly associated with lower overall mortality rates (HR 0.31, 95% CI 0.17 to 0.58; p < 0.001). Heart failure was also significantly associated with the long-term cardiac mortality (p = 0.032). Of all 303 patients, 20 had more than 1 disease episode. Chronic dialysis (p = 0.002), intravenous drug use (p = 0.002) and diabetes (p = 0.015) were significant risk factors for recurrent episodes of IE, but when analysed separately for the 1-year survivors, only chronic dialysis remained significant (p = 0.017). Recurrences and late valve surgery did not confer a poor prognosis. CONCLUSION: Heart failure during the index episode of IE was the complication, which significantly predicted a poor long-term outcome. Patients who underwent surgery during the initial hospitalisation for IE faired significantly better than those who did not. PMID- 18419813 TI - Breast cancer risk in relation to urinary and serum biomarkers of phytoestrogen exposure in the European Prospective into Cancer-Norfolk cohort study. AB - INTRODUCTION: Phytoestrogens are a group of compounds found in plants that structurally resemble the hormone oestradiol, and thus have the potential to act as oestrogen agonists or antagonists. Their potential effects may alter the risk of breast cancer, but only a limited range of phytoestrogens has been examined in prospective cohort studies. METHODS: Serum and urine samples from 237 incident breast cancer cases and 952 control individuals (aged 45 to 75 years) in the European Prospective into Cancer-Norfolk cohort were analysed for seven phytoestrogens (daidzein, enterodiol, enterolactone, genistein, glycitein, o desmethylangolensin, and equol) using liquid chromatography/mass spectrometry. Data on participants' diet, demographics, anthropometrics, and medical history were collected upon recruitment. All models were adjusted for weight, fat and energy intake, family history of breast cancer, social class, analytical batch, and factors related to oestrogen exposure. RESULTS: Urinary or serum phytoestrogens were not associated with protection from breast cancer in the European Prospective into Cancer-Norfolk cohort. Breast cancer risk was marginally increased with higher levels of total urinary isoflavones (odds ratio = 1.08 (95% confidence interval = 1.00 to 1.16), P = 0.055); among those with oestrogen receptor-positive tumours, the risk of breast cancer was increased with higher levels of urinary equol (odds ratio = 1.07 (95% confidence interval = 1.01 to 1.12), P = 0.013). CONCLUSION: There was limited evidence of an association between phytoestrogen biomarkers and breast cancer risk in the present study. There was no indication of decreased likelihood of breast cancer with higher levels of phytoestrogen biomarkers, but the observation that some phytoestrogen biomarkers may be associated with greater risk of breast cancer warrants further study with greater statistical power. PMID- 18419814 TI - Proteome alteration induced by hTERT transfection of human fibroblast cells. AB - BACKGROUND: Telomerase confers cellular immortality by elongating telomeres, thereby circumventing the Hayflick limit. Extended-life-span cells have been generated by transfection with the human telomerase reverse transcriptase (hTERT) gene. hTERT transfected cell lines may be of outstanding interest to monitor the effect of drugs targeting the telomerase activity. The incidence of hTERT gene transfection at the proteome level is a prerequisite to that purpose. The effect of the transfection has been studied on the proteome of human fibroblast (WI38). Cytosolic and nuclear fractions of WI38 cells, empty vector transfected WI38 (WI38-HPV) and hTERT WI38 cells were submitted to a 2D-DIGE (Two-Dimensional Differential In-Gel Electrophoresis) analysis. Only spots that had a similar abundance in WI38 and WI38-HPV, but were differentially expressed in WI38 hTERT were selected for MS identification. This method directly points to the proteins linked with the hTERT expression. Number of false positive differentially expressed proteins has been excluded by using control WI38-HPV cells. The proteome alteration induced by hTERT WI38 transfection should be taken into account in subsequent use of the cell line for anti-telomerase drugs evaluation. RESULTS: 2D-DIGE experiment shows that 57 spots out of 2246 are significantly differentially expressed in the cytosolic fraction due to hTERT transfection, and 38 were confidently identified. In the nuclear fraction, 44 spots out of 2172 were selected in the differential proteome analysis, and 14 were identified. The results show that, in addition to elongating telomeres, hTERT gene transfection has other physiological roles, among which an enhanced ER capacity and a potent cell protection against apoptosis. CONCLUSION: We show that the methodology reduces the complexity of the proteome analysis and highlights proteins implicated in other processes than telomere elongation. hTERT induced proteome changes suggest that telomerase expression enhances natural cell repair mechanisms and stress resistance probably required for long term resistance of immortalized cells. Thus, hTERT transfected cells can not be only consider as an immortal equivalent to parental cells but also as cells which are over-resistant to stresses. These findings are the prerequisite for any larger proteomics aiming to evaluate anti-telomerase drugs proteome alteration and thus therapeutics induced cell reactions. PMID- 18419815 TI - Erwin Schroedinger, Francis Crick and epigenetic stability. AB - Schroedinger's book 'What is Life?' is widely credited for having played a crucial role in development of molecular and cellular biology. My essay revisits the issues raised by this book from the modern perspective of epigenetics and systems biology. I contrast two classes of potential mechanisms of epigenetic stability: 'epigenetic templating' and 'systems biology' approaches, and consider them from the point of view expressed by Schroedinger. I also discuss how quantum entanglement, a nonclassical feature of quantum mechanics, can help to address the 'problem of small numbers' that led Schroedinger to promote the idea of a molecular code-script for explaining the stability of biological order. PMID- 18419816 TI - Efficacy of amodiaquine in the treatment of uncomplicated falciparum malaria in young children of rural north-western Burkina Faso. AB - BACKGROUND: Combination therapy has become a new paradigm in malaria treatment. Amodiaquine is a common partner drug in different malaria combination therapies used or investigated in sub-Saharan Africa, but data on its efficacy as a single drug are scarce. METHODS: The objective of the study was to determine the efficacy of amodiaquine against falciparum malaria in neighbouring rural and urban areas of north-western Burkina Faso. The study was designed as an uncontrolled trial in children aged 6-59 months with uncomplicated falciparum malaria in the Nouna Health District. RESULTS: During the rainy season 2005, 117 children were enrolled, 62 from the rural and 55 from the urban study area. The crude adequate clinical and parasitological response (ACPR) rate was 103/117 (88%) by day 14 but decreased to 28/117 (24%) by day 28. After PCR correction for reinfections, ACPR rates were 108/117 (92%) and 71/117 (61%) by day 14 and day 28, respectively. There were no significant differences in efficacy between urban and rural areas. The Plasmodium falciparum crt K76T mutation not predict AQ failure, but was selected in parasites re-appearing following treatment. No serious adverse events occurred and only 16 other adverse events were recorded. CONCLUSION: Compared to chloroquine, amodiaquine is more effective against uncomplicated falciparum malaria in Burkina Faso. However, a considerable degree of amodiaquine resistance already exists and it is currently unclear how this resistance will develop when amodiaquine in combination with other drugs is used on a large scale. TRIAL REGISTRATION: Current Controlled Trials ISRCTN73824458. PMID- 18419817 TI - Metabolic pathways of the wheat (Triticum aestivum) endosperm amyloplast revealed by proteomics. AB - BACKGROUND: By definition, amyloplasts are plastids specialized for starch production. However, a proteomic study of amyloplasts isolated from wheat (Triticum aestivum Butte 86) endosperm at 10 days after anthesis (DPA) detected enzymes from many other metabolic and biosynthetic pathways. To better understand the role of amyloplasts in food production, the data from that study were evaluated in detail and an amyloplast metabolic map was outlined. RESULTS: Analysis of 288 proteins detected in an amyloplast preparation predicted that 178 were amyloplast proteins. Criteria included homology with known plastid proteins, prediction of a plastid transit peptide for the wheat gene product or a close homolog, known plastid location of the pathway, and predicted plastid location for other members of the same pathway. Of these, 135 enzymes were arranged into 18 pathways for carbohydrate, lipid, amino acid, nucleic acid and other biosynthetic processes that are critical for grain-fill. Functions of the other proteins are also discussed. CONCLUSION: The pathways outlined in this paper suggest that amyloplasts play a central role in endosperm metabolism. The interacting effects of genetics and environment on starch and protein production may be mediated in part by regulatory mechanisms within this organelle. PMID- 18419818 TI - Extensive neuroadaptive changes in cortical gene-transcript expressions of the glutamate system in response to repeated intermittent MDMA administration in adolescent rats. AB - BACKGROUND: Many studies have focused on the implication of the serotonin and dopamine systems in neuroadaptive responses to the recreational drug 3,4 methylenedioxy-metamphetamine (MDMA). Less attention has been given to the major excitatory neurotransmitter glutamate known to be implicated in schizophrenia and drug addiction. The aim of the present study was to investigate the effect of repeated intermittent MDMA administration upon gene-transcript expression of the glutamate transporters (EAAT1, EAAT2-1, EAAT2-2), the glutamate receptor subunits of AMPA (GluR1, GluR2, GluR3), the glutamate receptor subunits of NMDA (NR1, NR2A and NR2B), as well as metabotropic glutamate receptors (mGluR1, mGluR2, mGluR3, mGluR5) in six different brain regions. Adolescent male Sprague Dawley rats received MDMA at the doses of 3 x 1 and 3 x 5 mg/kg/day, or 3x vehicle 3 hours apart, every 7th day for 4 weeks. The gene-transcript levels were assessed using real-time PCR validated with a range of housekeeping genes. RESULTS: The findings showed pronounced enhancements in gene-transcript expression of GluR2, mGluR1, mGluR5, NR1, NR2A, NR2B, EAAT1, and EAAT2-2 in the cortex at bregma +1.6. In the caudate putamen, mRNA levels of GluR3, NR2A, and NR2B receptor subunits were significantly increased. In contrast, the gene-transcript expression of GluR1 was reduced in the hippocampus. In the hypothalamus, there was a significant increase of GluR1, GluR3, mGluR1, and mGluR3 gene-transcript expressions. CONCLUSION: Repeated intermittent MDMA administration induces neuroadaptive changes in gene transcript expressions of glutamatergic NMDA and AMPA receptor subunits, metabotropic receptors and transporters in regions of the brain regulating reward related associative learning, cognition, and memory and neuro-endocrine functions. PMID- 18419819 TI - Non-invasive monitoring of tissue oxygenation during laparoscopic donor nephrectomy. AB - BACKGROUND: Standard methods for assessment of organ viability during surgery are typically limited to visual cues and tactile feedback in open surgery. However, during laparoscopic surgery, these processes are impaired. This is of particular relevance during laparoscopic renal donation, where the condition of the kidney must be optimized despite considerable manipulation. However, there is no in vivo methodology to monitor renal parenchymal oxygenation during laparoscopic surgery. METHODS: We have developed a method for the real time, in vivo, whole organ assessment of tissue oxygenation during laparoscopic nephrectomy to convey meaningful biological data to the surgeon during laparoscopic surgery. We apply the 3-CCD (charge coupled device) camera to monitor qualitatively renal parenchymal oxygenation with potential real-time video capability. RESULTS: We have validated this methodology in a porcine model across a range of hypoxic conditions, and have then applied the method during clinical laparoscopic donor nephrectomies during clinically relevant pneumoperitoneum. 3-CCD image enhancement produces mean region of interest (ROI) intensity values that can be directly correlated with blood oxygen saturation measurements (R2 > 0.96). The calculated mean ROI intensity values obtained at the beginning of the laparoscopic nephrectomy do not differ significantly from mean ROI intensity values calculated immediately before kidney removal (p > 0.05). CONCLUSION: Here, using the 3-CCD camera, we qualitatively monitor tissue oxygenation. This means of assessing intraoperative tissue oxygenation may be a useful method to avoid unintended ischemic injury during laparoscopic surgery. Preliminary results indicate that no significant changes in renal oxygenation occur as a result of pneumoperitoneum. PMID- 18419820 TI - An assessment of technology-based service encounters & network security on the e health care systems of medical centers in Taiwan. AB - BACKGROUND: Enhancing service efficiency and quality has always been one of the most important factors to heighten competitiveness in the health care service industry. Thus, how to utilize information technology to reduce work load for staff and expeditiously improve work efficiency and healthcare service quality is presently the top priority for every healthcare institution. In this fast changing modern society, e-health care systems are currently the best possible way to achieve enhanced service efficiency and quality under the restraint of healthcare cost control. The electronic medical record system and the online appointment system are the core features in employing e-health care systems in the technology-based service encounters. METHODS: This study implemented the Service Encounters Evaluation Model, the European Customer Satisfaction Index, the Attribute Model and the Overall Affect Model for model inference. A total of 700 copies of questionnaires from two authoritative southern Taiwan medical centers providing the electronic medical record system and the online appointment system service were distributed, among which 590 valid copies were retrieved with a response rate of 84.3%. We then used SPSS 11.0 and the Linear Structural Relationship Model (LISREL 8.54) to analyze and evaluate the data. RESULTS: The findings are as follows: (1) Technology-based service encounters have a positive impact on service quality, but not patient satisfaction; (2) After experiencing technology-based service encounters, the cognition of the service quality has a positive effect on patient satisfaction; and (3) Network security contributes a positive moderating effect on service quality and patient satisfaction. CONCLUSION: It revealed that the impact of electronic workflow (online appointment system service) on service quality was greater than electronic facilities (electronic medical record systems) in technology-based service encounters. Convenience and credibility are the most important factors of service quality in technology-based service encounters that patients demand. Due to the openness of networks, patients worry that transaction information could be intercepted; also, the credibility of the hospital involved is even a bigger concern, as patients have a strong sense of distrust. Therefore, in the operation of technology-based service encounters, along with providing network security, it is essential to build an atmosphere of psychological trust. PMID- 18419821 TI - Ornithine-delta-aminotransferase is essential for arginine catabolism but not for proline biosynthesis. AB - BACKGROUND: Like many other plant species, Arabidopsis uses arginine (Arg) as a storage and transport form of nitrogen, and proline (Pro) as a compatible solute in the defence against abiotic stresses causing water deprivation. Arg catabolism produces ornithine (Orn) inside mitochondria, which was discussed controversially as a precursor for Pro biosynthesis, alternative to glutamate (Glu). RESULTS: We show here that ornithine-delta-aminotransferase (deltaOAT, At5g46180), the enzyme converting Orn to pyrroline-5-carboxylate (P5C), is localised in mitochondria and is essential for Arg catabolism. Wildtype plants could readily catabolise supplied Arg and Orn and were able to use these amino acids as the only nitrogen source. Deletion mutants of deltaOAT, however, accumulated urea cycle intermediates when fed with Arg or Orn and were not able to utilize nitrogen provided as Arg or Orn. Utilisation of urea and stress induced Pro accumulation were not affected in T-DNA insertion mutants with a complete loss of deltaOAT expression. CONCLUSION: Our findings indicate that deltaOAT feeds P5C exclusively into the catabolic branch of Pro metabolism, which yields Glu as an end product. Conversion of Orn to Glu is an essential route for recovery of nitrogen stored or transported as Arg. Pro biosynthesis occurs predominantly or exclusively via the Glu pathway in Arabidopsis and does not depend on Glu produced by Arg and Orn catabolism. PMID- 18419822 TI - The LIM and SH3 domain protein family: structural proteins or signal transducers or both? AB - LIM and SH3 Protein 1 (LASP-1) was initially identified from a cDNA library of metastatic axillary lymph nodes (MLN) more than a decade ago. It was found to be overexpressed in human breast and ovarian cancer and became the first member of a newly defined LIM-protein subfamily of the nebulin group characterized by the combined presence of LIM and SH3 domains. LASP2, a novel LASP1-related gene was first identified and characterized in silico. Subsequently it proved to be a splice variant of the Nebulin gene and therefore was also termed LIM/nebulette. LASP-1 and -2 are highly conserved in their LIM, nebulin-like and SH3 domains but differ significantly at their linker regions. Both proteins are ubiquitously expressed and involved in cytoskeletal architecture, especially in the organization of focal adhesions. Here we present the first systematic review to summarize all relevant data concerning their domain organization, expression profiles, regulating factors and function. We compile evidence that both, LASP-1 and LASP-2, are important during early embryo- and fetogenesis and are highly expressed in the central nervous system of the adult. However, only LASP-1 seems to participate significantly in neuronal differentiation and plays an important functional role in migration and proliferation of certain cancer cells while the role of LASP-2 is more structural. The increased expression of LASP-1 in breast tumours correlates with high rates of nodal-metastasis and refers to a possible relevance as a prognostic marker. PMID- 18419823 TI - Initial intramuscular perfusion pressure predicts early skeletal muscle function following isolated tibial fractures. AB - BACKGROUND: The severity of associated soft tissue trauma in complex injuries of the extremities guides fracture treatment and decisively determines patient's prognosis. Trauma-induced microvascular dysfunction and increased tissue pressure is known to trigger secondary soft tissue damage and seems to adversely affect skeletal muscle function. METHODS: 20 patients with isolated tibial fractures were included. Blood pressure and compartment pressure (anterior and deep posterior compartment) were measured continuously up to 24 hours. Corresponding perfusion pressure was calculated. After 4 and 12 weeks isokinetic muscle peak torque and mean power of the ankle joint in dorsal and plantar flexion were measured using a Biodex dynamometer. RESULTS: A significant inverse correlation between the anterior perfusion pressure at 24 hours and deficit in dorsiflexion at 4 weeks was found for both, the peak torque (R = -0.83; p < 0.01) and the mean power (R = -0.84; p < 0.01). The posterior perfusion pressure at 24 h and the plantar flexion after 4 weeks in both, peak torque (R = -0.73, p = or < 0.05) and mean power (R = -0.7, p = or < 0.05) displayed a significant correlation. CONCLUSION: The functional relationship between the decrease in intramuscular perfusion pressures and muscle performance in the early rehabilitation period indicate a causative and prognostic role of early posttraumatic microcirculatory derangements and skeletal muscle function. Therapeutic concepts aimed at effective muscle recovery, early rehabilitation, and decreased secondary tissue damage, should consider the maintenance of an adequate intramuscular perfusion pressure. PMID- 18419824 TI - Prolonged gastroparesis after corrective surgery for Wilkie's syndrome: a case report. AB - INTRODUCTION: Wilkie's syndrome, a rare cause of intestinal obstruction, is related to anatomical and mechanical factors associated with the reduction of retroperitoneal fat padding. The diagnostic challenges of identifying vascular constriction between the aorta and superior mesenteric artery have been answered by advances in the field of computed tomography. Despite diagnostic confusion with intestinal dysmotility syndrome, conservative therapy with nutritional supplementation is the initial approach and duodenojejunostomy is favoured if non surgical treatment fails. CASE PRESENTATION: We present a case of a 49-year-old woman with Wilkie's syndrome with persistent symptoms of gastroparesis for 15 months following corrective surgery. CONCLUSION: Open and laparoscopic duodenojejunostomy have been described as the best surgical treatment options for Wilkie's syndrome, but further work needs to be done for patients with refractory symptoms of gastroparesis after these corrective surgeries. PMID- 18419825 TI - Effects of resuscitation with crystalloid fluids on cardiac function in patients with severe sepsis. AB - BACKGROUND: The use of hypertonic crystalloid solutions, including sodium chloride and bicarbonate, for treating severe sepsis has been much debated in previous investigations. We have investigated the effects of three crystalloid solutions on fluid resuscitation in severe sepsis patients with hypotension. METHODS: Ninety-four severe sepsis patients with hypotension were randomly assigned to three groups. The patients received the following injections within 15 min at initial treatment: Ns group (n = 32), 5 ml/kg normal saline; Hs group (n = 30), with 5 ml/kg 3.5% sodium chloride; and Sb group (n = 32), 5 ml/kg 5% sodium bicarbonate. Cardiac output (CO), systolic blood pressure, mean arterial pressure (MAP), body temperature, heart rate, respiratory rate and blood gases were measured. RESULTS: There were no differences among the three groups in CO, MAP, heart rate or respiratory rate during the 120 min trial or the 8 hour follow up, and no significant differences in observed mortality rate after 28 days. However, improvement of MAP and CO started earlier in the Sb group than in the Ns and Hs groups. Sodium bicarbonate increased the base excess but did not alter blood pH, lactic acid or [HCO3]- values; and neither 3.5% hypertonic saline nor 5% sodium bicarbonate altered the Na+, K+, Ca2+ or Cl- levels. CONCLUSION: All three crystalloid solutions may be used for initial volume loading in severe sepsis, and sodium bicarbonate confers a limited benefit on humans with severe sepsis. TRIAL REGISTRATION: ISRCTN36748319. PMID- 18419826 TI - The calcium channel beta2 (CACNB2) subunit repertoire in teleosts. AB - BACKGROUND: Cardiomyocyte contraction is initiated by influx of extracellular calcium through voltage-gated calcium channels. These oligomeric channels utilize auxiliary beta subunits to chaperone the pore-forming alpha subunit to the plasma membrane, and to modulate channel electrophysiology 1. Several beta subunit family members are detected by RT-PCR in the embryonic heart. Null mutations in mouse beta2, but not in the other three beta family members, are embryonic lethal at E10.5 due to defects in cardiac contractility 2. However, a drawback of the mouse model is that embryonic heart rhythm is difficult to study in live embryos due to their intra-uterine development. Moreover, phenotypes may be obscured by secondary effects of hypoxia. As a first step towards developing a model for contributions of beta subunits to the onset of embryonic heart rhythm, we characterized the structure and expression of beta2 subunits in zebrafish and other teleosts. RESULTS: Cloning of two zebrafish beta2 subunit genes (beta2.1 and beta2.2) indicated they are membrane-associated guanylate kinase (MAGUK) family genes. Zebrafish beta2 genes show high conservation with mammals within the SH3 and guanylate kinase domains that comprise the "core" of MAGUK proteins, but beta2.2 is much more divergent in sequence than beta2.1. Alternative splicing occurs at the N-terminus and within the internal HOOK domain. In both beta2 genes, alternative short ATG-containing first exons are separated by some of the largest introns in the genome, suggesting that individual transcript variants could be subject to independent cis-regulatory control. In the Tetraodon nigrovidis and Fugu rubripes genomes, we identified single beta2 subunit gene loci. Comparative analysis of the teleost and human beta2 loci indicates that the short 5' exon sequences are highly conserved. A subset of 5' exons appear to be unique to teleost genomes, while others are shared with mammals. Alternative splicing is temporally and spatially regulated in embryo and adult. Moreover, a different subset of spliced beta2 transcript variants is detected in the embryonic heart compared to the adult. CONCLUSION: These studies refine our understanding of beta2 subunit diversity arising from alternative splicing, and provide the groundwork for functional analysis of beta2 subunit diversity in the embryonic heart. PMID- 18419827 TI - Molecular and cellular limits to somatosensory specificity. AB - Animals detect environmental changes through sensory neural mechanisms that enable them to differentiate the quality, intensity and temporal characteristics of stimuli. The 'doctrine of specific nervous energies' postulates that the different sensory modalities experienced by humans result of the activation of specific nervous pathways. Identification of functional classes of sensory receptors provided scientific support to the concept that somatosensory modalities (touch, pain, temperature, kinesthesis) are subserved by separate populations of sensory receptor neurons specialized in detecting innocuous and injurious stimuli of different quality (mechanical forces, temperature, chemical compounds). The identification of receptor proteins activated by different physicochemical stimuli, in particular ion channels of the Transient Receptor Potential (TRP) superfamily, has put forward the concept that specificity of peripheral sensory receptor neurons is determined by their expression of a particular "molecular sensor" that confers to each functional type its selectivity to respond with a discharge of nerve impulses to stimuli of a given quality. Nonetheless, recent experimental data suggest that the various molecular sensors proposed as specific transducer molecules for stimuli of different quality are not as neatly associated with the distinct functional types of sensory receptors as originally proposed. First, many ion channel molecules initially associated to the transduction of only one particular form of energy are also activated by stimuli of different quality, implying a limited degree of specificity in their transducing capacities. Second, molecular sensors associated with a stimulus quality and hence to a sensory receptor type and ultimately to a sensory modality may be concomitantly expressed in sensory receptor neurons functionally defined as specific for another stimulus quality. Finally, activation of voltage gated channels involved primarily in nerve impulse generation can also influence the gating of transducing channels, dramatically modifying their activation profile. Thus, we propose that the capacity exhibited by the different functional types of somatosensory receptor neurons to preferentially detect and encode specific stimuli into a discharge of nerve impulses, appears to result of a characteristic combinatorial expression of different ion channels in each neuronal type that finally determines their transduction and impulse firing properties. Transduction channels don't operate in isolation and their cellular context should also be taken into consideration to fully understand their function. Moreover, the inhomogeneous distribution of transduction and voltage-gated channels at soma, axonal branches and peripheral endings of primary sensory neurons influences the characteristics of the propagated impulse discharge that encodes the properties of the stimulus. Alteration of this concerted operation of ion channels in pathological conditions may underlie the changes in excitability accompanying peripheral sensory neuron injuries. PMID- 18419828 TI - Mucosal administration of alpha-fodrin inhibits experimental Sjogren's syndrome autoimmunity. AB - INTRODUCTION: alpha-Fodrin is an autoantigen in Sjogren's syndrome. We hypothesized that mucosal administration of alpha-fodrin might prevent the disease. METHODS: Four-week-old NOD mice were immunized (intranasal) with a 1 microg or 10 microg dose of alpha-fodrin every other day. PBS 10 microl/dose and Glutathione transferase (GST 10 microg/dose (control mice) were intranasally administrated by the same procedure. The salivary flow was maintained in immunized animals. The animals were analyzed for the presence of anti-Sjogren's syndrome A, anti-Sjogren's syndrome B, rheumatoid factor and antinuclear, anti alpha-fodrin, and anti-type 3 muscarinic acetylcholine receptor polypeptide (anti M3RP) by immunofluorescence or ELISA. The cytokines IFNgamma and IL-10 were measured by ELISA. Salivary glands were examined by H&E staining and immunohistochemical analysis. The water-volume intake was calculated for each group. The induction of regulatory T cells was assessed by fluorescence-activated cell sorting analysis for the frequency of Foxp3+ cells among peripheral CD4+CD25+ T cells. RESULTS: The appearance of anti-alpha-fodrin and anti-M3RP antibodies was delayed in mice immunized with alpha-fodrin. The titers of anti alpha-fodrin and anti-M3RP antibodies were lower in immunized mice (P < 0.05), but there was no significant difference between the low-dose or high-dose immunization groups. Five out of eight mice in the GST group, five of eight mice in the PBS group, two of eight mice in the alpha-fodrin 1 microg/dose group, and three out of eight mice in the alpha-fodrin 10 microg/dose were positive for antinuclear antibodies. The levels of serum IFNgamma in mice immunized with 1 microg/dose or 10 microg/dose alpha-fodrin, with PBS, and with GST were 41.9 +/- 16.2 pg/ml, 37.1 +/- 15.4 pg/ml, 86.8 +/- 17.8 pg/ml and 71.6 +/- 11.1 pg/ml, respectively, while we found no difference in the levels of serum IL-10 among the groups. The number of Foxp3+ CD4+CD25+ regulatory T cells was higher in the alpha fodrin groups compared with the PBS and GST control groups (P < 0.05). Lymphocytic infiltration and expression of alpha-fodrin in the salivary glands was decreased in alpha-fodrin-treated groups. The fluid intake of mice in the 1 microg/dose alpha-fodrin, 10 microg/dose alpha-fodrin, PBS, and GST groups was 39.2 +/- 2.1 ml, 40.4 +/- 2.5 ml, 49.3 +/- 3.1 ml and 51.6 +/- 2.8 ml, respectively. CONCLUSION: Mucosal administration of alpha-fodrin effectively inhibited the progression of experimental Sjogren's syndrome autoimmunity. PMID- 18419829 TI - Neuroleptic-induced movement disorders in a naturalistic schizophrenia population: diagnostic value of actometric movement patterns. AB - BACKGROUND: Neuroleptic-induced movement disorders (NIMDs) have overlapping co morbidity. Earlier studies have described typical clinical movement patterns for individual NIMDs. This study aimed to identify specific movement patterns for each individual NIMD using actometry. METHODS: A naturalistic population of 99 schizophrenia inpatients using conventional antipsychotics and clozapine was evaluated. Subjects with NIMDs were categorized using the criteria for NIMD found in the Diagnostic and Statistical Manual for Mental Disorders - Fourth Edition (DSM-IV).Two blinded raters evaluated the actometric-controlled rest activity data for activity periods, rhythmical activity, frequencies, and highest acceleration peaks. A simple subjective question was formulated to test patient based evaluation of NIMD. RESULTS: The patterns of neuroleptic-induced akathisia (NIA) and pseudoakathisia (PsA) were identifiable in actometry with excellent inter-rater reliability. The answers to the subjective question about troubles with movements distinguished NIA patients from other patients rather well. Also actometry had rather good screening performances in distinguishing akathisia from other NIMD. Actometry was not able to reliably detect patterns of neuroleptic induced parkinsonism and tardive dyskinesia. CONCLUSION: The present study showed that pooled NIA and PsA patients had a different pattern in lower limb descriptive actometry than other patients in a non-selected sample. Careful questioning of patients is a useful method of diagnosing NIA in a clinical setting. PMID- 18419830 TI - Stem-cell-abundant proteins Nanog, Nucleostemin and Musashi1 are highly expressed in malignant cervical epithelial cells. AB - BACKGROUND: Nanog, nucleostemin (NS) and musashi1 (Msi1) are proteins that are highly expressed in undifferentiated embryonic stem (ES) cells and have been shown to be essential in maintaining the pluripotency and regulating the proliferation and asymmetric division of ES cells and several nervous system tumor cells. The roles of Nanog, NS and Msi1 in development and progression of cervical carcinoma have, until now, not been well documented. METHODS: In this study, expression of Nanog, NS and Msi1 was detected by immunohistochemistry analysis in 235 patients with various degrees of cervical epithelial lesions, including 49 with normal cervical epithelia, 31 with mild dysplasia (CIN I), 77 with moderate-severe dysplasia (CIN II-III) and 78 with squamous cervical carcinomas (SCCs). Associations with various clinical pathological prognostic variables were analyzed in 50 early-stage SCC patients. RESULTS: Nanog, NS and Msi1 expression levels were significantly higher in SCC patients compared with CIN patients, and were higher in CIN patients compared with those with normal cervical epithelia. Nanog expression levels showed significantly differences according to different tumor sizes (P < 0.05), whereas there were no differences in NS and Msi1 expression levels according to different clinical pathological parameters. CONCLUSION: Our findings indicate that Nanog, NS and Msi1 may be involved in carcinogenesis of the cervix and progression of cervical carcinoma. PMID- 18419831 TI - Prenatal diagnosis in the first trimester of pregnancy. AB - In the past 2 decades, the second trimester of pregnancy has been the most common time for prenatal diagnosis of fetal anomalies and chromosomal aneuploidies. More recently, screening for and diagnosis of chromosomal abnormalities are increasingly being performed in the first trimester. With improvements and technological advances in ultrasound, it is now possible to identify many fetal structural anomalies at 11 to 13 6/7 weeks' gestation. At 10 to 11 weeks' gestation, biochemical markers in serum-PAPP-A, free beta-hCG, AFP, and uE3 combined with sonographic measurement of nuchal translucency and the presence/absence of the nasal bone can achieve a Down syndrome detection rate of 97.5% at a false-positive rate of 5%. Structural anomalies of the central nervous system, and the cardiac, renal, and gastrointestinal tracts can now be diagnosed by either transabdominal or transvaginal scanning, achieving detection of up to 80% of CNS anomalies by 13 weeks' gestation. In future, the emphasis in prenatal diagnosis will likely be in the first trimester. TARGET AUDIENCE: Obstetricians & Gynecologists, Family Physicians. LEARNING OBJECTIVES: After completion of this article, the reader should be able to explain the rationale for first trimester combined ultrasound and serum analyte screening for fetal Down syndrome, describe the fetal anatomic structures that can be seen and evaluated in the first trimester, provide patient counseling about the relative benefits of genetic amniocentesis versus chorionic villous sampling, and discuss the application of Doppler technology to the evaluation of a first trimester fetus. PMID- 18419832 TI - Effective medical education: insights from the Cochrane Library. AB - In 2006, the Accreditation Council for Continuing Medical Education highlighted the need for linking educational activities to changes in competence, performance, or patient outcomes. Hence, educational providers increasingly need to know what strategies are effective. The Cochrane Library is widely regarded as the best source of credible evidence concerning health care. The authors searched the Cochrane Database of Systematic Reviews (issue 4 for 2006) using the search terms "continuing medical education," "medical education," and "continuing education." They conducted a second complementary search of this database by review group (Effective Practice and Organization of Care). Finally, the authors examined the references of recent review articles for Cochrane reviews and found 9 relevant reviews. The most effective educational methods were the most interactive. Combined didactic presentations and workshops were more effective than traditional didactic presentations alone. Medical education was more effective when more than 1 intervention occurred, especially if these interventions occurred over an extended period. Targeted education should focus on changing a behavior that is simple, because effect size is inversely proportional to the complexity of the behavior. In the era of evidence-based medicine, interventions-including educational ones-should reflect the best available evidence. Cochrane reviews of randomized controlled trials of educational methods provide important guidance that often challenges traditional didactic approaches. Integrating the findings from the Cochrane reviews may allow continuing medical education to be more successful in bringing about changes to healthcare providers' behavior. TARGET AUDIENCE: Obstetricians & Gynecologists, Family Physicians. LEARNING OBJECTIVES: After completion of this article, the reader should be able to explain the scientific evidence concerning the effectiveness of various techniques used for continuing medical education, state the relative value of such techniques as traditional didactic lectures, conferences led by local opinion leaders, interactive workshops, and educational outreach visits, and identify the value and limitations of teaching critical appraisal skills. PMID- 18419833 TI - Fertility and impact of pregnancies on the mother and child in classic galactosemia. AB - Despite the high prevalence of premature ovarian failure (POF) and subsequent infertility in galactosemic women, spontaneous pregnancies occur and may not be as rare as is generally assumed. This is important for counseling these women on fertility. The purpose of this review is to assess the occurrence and predicting factors of pregnancy, and to evaluate the impact of pregnancy on the mother's and child's health. The female Dutch galactosemia population (age > 18 years) was studied, and a literature search on articles reporting pregnancy in galactosemic women, published between January 1971 and December 2007, was performed. Twenty two galactosemic women were studied. Nine women have tried to conceive, of which 4 were successful. Three mothers were diagnosed with POF before the first pregnancy and/or in between pregnancies. In literature, 50 pregnancy reports were found. In 10 pregnancy reports from the literature, the mother's genotype is known. Four women were homozygous for the Q188R mutation, which equals the incidence of 40-45% of classic galactosemia caused by this mutation. This study challenges the current opinion that the chance of becoming pregnant is small in classic galactosemia. Despite POF in most galactosemic women, pregnancies do occur. The genotype and GALT-activity do not seem to predict the chance of becoming pregnant, whereas the occurrence of spontaneous menarche might. No evidence for the need of additional check-ups during the pregnancy and puerperium was found. Elevations in galactose-metabolites do occur, but without evidence of clinical impact for the mother or the child, although possible long-term effects have not been thoroughly investigated. TARGET AUDIENCE: Obstetricians & Gynecologists, Family Physicians. LEARNING OBJECTIVES: After completion of this article, the reader should be able to summarize the purported causes and sequelae of galactosemia, explain the possible sequelae of galactosemia, distinguish alterations of the ovary and the hypothalamic-pituitary axis, identify the frequency of pregnancy and the possible outcome of the offspring, and outline dietary management of patients with galactosemia. PMID- 18419834 TI - The basal ganglia circuits, dopamine, and ambiguous word processing: a neurobiological account of priming studies in Parkinson's disease. AB - Research into the processing of lexical ambiguities has provided a valuable paradigm for investigating the functional architecture of the language processing system in normal and neurologically impaired populations and specifically, how basal ganglia circuits and the neurotransmitter dopamine may act to enhance and/or suppress various meanings relative to the context in which the lexical ambiguity appears. In this review, we develop the hypothesis that an integrated basal ganglia thalamocortical circuit linking the striatum and inferior frontal cortex is involved in the enhancement and suppression of ambiguous word meanings when a lexical ambiguity is presented within a linguistic context. Reference to behavioral, neurophysiological, and neurochemical studies of subcortical function in both healthy populations and people with Parkinson's disease will be used to provide further support for the proposal that the subcortex is integrally involved in ambiguous word processing. PMID- 18419835 TI - Not all repetition is alike: different benefits of repetition in amnesia and normal memory. AB - While it is well known that repetition can enhance memory in amnesia, little is known about which forms of repetition are most beneficial. This study compared the effect on recognition memory of repetition of words in the same semantic context and in varied semantic contexts. To gain insight into the mechanisms by which these forms of repetition affect performance, participants were asked to make Remember/Know judgments during recognition. These judgments were used to make inferences about the contribution of recollection and familiarity to performance. For individuals with intact memory, the two forms of repetition were equally beneficial to overall recognition, and were associated with both enhanced Remember and Know responses. However, varied repetition was associated with a higher likelihood of Remember responses than was fixed repetition. The two forms of repetition also conferred equivalent benefits on overall recognition in amnesia, but in both cases, this enhancement was manifest exclusively in enhanced Know responses. We conclude that the repetition of information, and especially repetition in varied contexts, enhances recollection in individuals with intact memory, but exclusively affects familiarity in patients with severe amnesia. PMID- 18419837 TI - Thalamic atrophy and cognition in unilateral temporal lobe epilepsy. AB - This study examined quantitative magnetic resonance volumes of the thalamus and hippocampus and determined their relationship with cognitive function and clinical seizure characteristics in a sample of 46 unilateral temporal lobe epilepsy (TLE) subjects (20 left and 26 right) and 29 controls. The hippocampus and thalamus exhibited different patterns of volume abnormality, different associations with clinical seizure characteristics, and different patterns of relationship with cognitive measures. Hippocampal volume reduction was primarily ipsilateral to the seizure focus, and thalamic volume reduction was bilateral. Ipsilateral hippocampal volume was significantly correlated with both early neurodevelopmental features (age of seizure onset) and disease characteristics (duration of epilepsy), whereas thalamus integrity was related only to disease variables. Hippocampal volume showed a selective association with verbal memory performance. In contrast, both left and right thalamic volumes were significantly correlated with performance on both memory and nonmemory cognitive domains. These findings underscore the importance of thalamic atrophy in chronic TLE and its potential implications for cognition. PMID- 18419836 TI - Distinct cognitive profiles and rates of decline on the Mattis Dementia Rating Scale in autopsy-confirmed frontotemporal dementia and Alzheimer's disease. AB - Neuropsychological studies have shown that patients with Frontotemporal dementia (FTD) perform worse than patients with Alzheimer's disease (AD) on tests of conceptualization and verbal fluency, but better on tests of memory and visuospatial functions. However, it is not known if these distinct cognitive profiles are robust enough to be detected using a relatively brief dementia screening instrument such as the Mattis Dementia Rating Scale (MDRS). To address this issue, the MDRS subscale profiles of patients with autopsy-confirmed FTD (n = 17) or AD (n = 34) were compared. Results showed distinct cognitive profiles in which FTD patients performed worse than AD patients on the Initiation/Perseveration and Conceptualization subscales while performing better on the Memory and Construction subscales. The distinct subscale profiles correctly classified 85% of AD patients and 76% of FTD patients. Profiles were maintained in a subset of mildly-to-moderately demented patients (MDRS > or = 105) and correctly classified 89% of these patients. In addition, FTD patients (mean = 30.0 points/year) declined faster than AD patients (mean = 14.8 points/year) on MDRS total and specific subscale scores. These results suggest that the MDRS may be a useful adjunct to other clinical measures for distinguishing FTD from AD and tracking the progression of the disorder. PMID- 18419838 TI - Differential neuropsychological test sensitivity to left temporal lobe epilepsy. AB - We examined the sensitivity of the Rey Auditory Verbal Learning Test (AVLT), California Verbal Learning Test (CVLT), Boston Naming Test (BNT), and Multilingual Aphasia Examination Visual Naming subtest (MAE VN) to lateralized temporal lobe epilepsy (TLE) in patients who subsequently underwent anterior temporal lobectomy. For the AVLT (n = 189), left TLE patients performed more poorly than their right TLE counterparts [left TLE = 42.9 (10.6), right TLE = 47.7 (9.9); p < .002 (Cohen's d = .47)]. Although statistically significant, the CVLT group difference (n = 212) was of a smaller magnitude [left LTE = 40.7 (11.1), right TLE = 43.8 (9.9); (p < .03, Cohen's d = .29)] than the AVLT. Group differences were also present for both measures of confrontation naming ability [BNT: left LTE = 43.1 (8.9), right TLE = 48.1 (8.9); p < .001 (Cohen's d = .56); MAE VN: left TLE = 42.2, right TLE = 45.6, p = .02 (Cohen's d = .36)]. When these data were modeled in independent logistic regression analyses, the AVLT and BNT both significantly predicted side of seizure focus, although the positive likelihood ratios were modest. In the subset of 108 patients receiving both BNT and AVLT, the AVLT was the only significant predictor of seizure laterality, suggesting individual patient variability regarding whether naming or memory testing may be more sensitive to lateralized TLE. PMID- 18419839 TI - Quantitative magnetic resonance image analysis of the cerebellum in macrocephalic and normocephalic children and adults with autism. AB - A detailed morphometric analysis of the cerebellum in autism with and without macrocephaly. Four subject groups (N = 65; male; IQs > or = 65; age 7 to 26 years) were studied with quantitative MRI; normocephalic and macrocephalic individuals with autism without mental retardation were compared to normocephalic and benign macrocephalic typically developing individuals. Total cerebellum volumes and surface areas of four lobular midsagittal groups were measured. Independent t-tests between autism and control subjects matched for head size revealed no significant differences. Multivariate analyses of variance were also performed, using the diagnostic group as the fixed factor, cerebellar measures as the dependent variables and total intracranial volume, total brain volume, age, verbal IQ, and performance IQ as covariates. No significant differences were found; however, a trend was noted in which macrocephalic individuals with autism consistently exhibited slightly smaller cerebellar volume or surface area when compared to individuals with benign macrocephaly. In autism, with and without macrocephaly, cerebellar structures were found to be proportional to head size and did not differ from typically developing subjects. PMID- 18419840 TI - Regional atrophy of the corpus callosum in dementia. AB - The regional distribution of degeneration of the corpus callosum (CC) in dementia is not yet clear. This study compared regional CC size in participants (n = 179) from the Cache County Memory and Aging Study. Participants represented a range of cognitive function: Alzheimer's disease (AD), vascular dementia (VaD), mild ambiguous (MA-cognitive problems, but not severe enough for diagnosis of dementia), and healthy older adults. CC outlines obtained from midsagittal magnetic resonance images were divided into 99 equally spaced widths. Factor analysis of these callosal widths identified 10 callosal regions. Multivariate analysis of variance revealed significant group differences for anterior and posterior callosal regions. Post-hoc pairwise comparisons of CC regions in patient groups as compared to the control group (controlling for age) revealed trends toward smaller anterior and posterior regions, but not all were statistically significant. As compared to controls, significantly smaller anterior and posterior CC regions were found in the AD group; significantly smaller anterior CC regions in the VaD group; but no significant CC regional differences in the MA group. Findings suggest that dementia-related CC atrophy occurs primarily in the anterior and posterior portions. PMID- 18419841 TI - Long-term neural processing of attention following early childhood traumatic brain injury: fMRI and neurobehavioral outcomes. AB - Attentional deficits are common and significant sequelae of pediatric traumatic brain injury (TBI). However, little is known about how the underlying neural processes that support different components of attention are affected. The present study examined brain activation patterns using fMRI in a group of young children who sustained a TBI in early childhood (n = 5; mean age = 9.4), and a group of age-matched control children with orthopedic injuries (OI) (n = 8) during a continuous performance task (CPT). Four children in the TBI group had moderate injuries, and one had a severe injury. Performance on the CPT task did not differ between groups. Both TBI and OI children activated similar networks of brain regions relevant to sustained attention processing, but the TBI group demonstrated several areas of significantly greater activation relative to controls, including frontal and parietal regions. These findings of over activation of the relevant attention network in the TBI group contrast with those obtained in imaging studies of Attention-Deficit/Hyperactivity Disorder where under-activation of the attention network has been documented. This study provides evidence that young children's brains function differently following a traumatic brain injury, and that these differences persist for years after the injury. PMID- 18419842 TI - Frequency and bases of abnormal performance by healthy adults on neuropsychological testing. AB - The frequency and determinants of abnormal test performance by normal individuals are critically important to clinical inference. Here we compare two approaches to predicting rates of abnormal test performance among healthy individuals with the rates actually shown by 327 neurologically normal adults aged 18-92 years. We counted how many participants produced abnormal scores, defined by three different cutoffs with test batteries of varied length, and the number of abnormal scores they produced. Observed rates generally were closer to predictions based on a series of Monte Carlo simulations than on the binomial model. They increased with the number of tests administered, decreased as more stringent cutoffs were used to identify abnormality, varied with the degree of correlation among test scores, and depended on individual differences in age, education, race, sex, and estimated premorbid IQ. Adjusting scores for demographic variables and premorbid IQ did not reduce rates of abnormal performance. However, it eliminated the contribution of these variables to rates of abnormal test performance. These findings raise fundamental questions about the nature and interpretation of abnormal test performance by normal, healthy adults. PMID- 18419843 TI - Visual perception in prediagnostic and early stage Huntington's disease. AB - Disturbances of visual perception frequently accompany neurodegenerative disorders but have been little studied in Huntington's disease (HD) gene carriers. We used psychophysical tests to assess visual perception among individuals in the prediagnostic and early stages of HD. The sample comprised four groups, which included 201 nongene carriers (NG), 32 prediagnostic gene carriers with minimal neurological abnormalities (PD1); 20 prediagnostic gene carriers with moderate neurological abnormalities (PD2), and 36 gene carriers with diagnosed HD. Contrast sensitivity for stationary and moving sinusoidal gratings, and tests of form and motion discrimination, were used to probe different visual pathways. Patients with HD showed impaired contrast sensitivity for moving gratings. For one of the three contrast sensitivity tests, the prediagnostic gene carriers with greater neurological abnormality (PD2) also had impaired performance as compared with NG. These findings suggest that early stage HD disrupts visual functions associated with the magnocellular pathway. However, these changes are only observed in individuals diagnosed with HD or who are in the more symptomatic stages of prediagnostic HD. PMID- 18419844 TI - Oralmotor slowing in multiple sclerosis: relationship to neuropsychological tasks requiring an oral response. AB - Although most neuropsychological batteries used with multiple sclerosis (MS) patients now exclude tests that require significant motor writing or manual manipulation speed, many of the most sensitive commonly used cognitive tests nonetheless require some type of rapid oral motor response. The aim of this study is to examine the extent to which primary oral motor articulation speed problems of individuals with MS contribute to performance and group differences on neuropsychological tasks requiring a rapid spoken response. Fifty MS patients and 50 healthy controls were administered the PASAT, COWAT, Animal Naming, and SDMT tests, in addition to a measure of rudimentary oral motor speed known as the maximum repetition rate of syllables and multisyllabic combinations (MRR) task. Regression analyses revealed that the amount of variance accounted for by the group (MS-Control) variable was reduced the following amounts for the tasks when the MRR was entered before the group variable: SDMT, 10% to 6%; PASAT, 4% to 2%; COWAT, 5% to 2%; Animal Naming, 11% to 7%. Our data suggest that rudimentary oral motor speed is slowed in MS patients and makes an important contribution to group differences in performance on commonly used neuropsychological tasks requiring a rapid spoken response. PMID- 18419845 TI - Potential for misclassification of mild cognitive impairment: a study of memory scores on the Wechsler Memory Scale-III in healthy older adults. AB - The psychometric criterion of mild cognitive impairment (MCI) generally involves having an unusually low score on memory testing (i.e., -1.5 SDs). However, healthy older adults can obtain low scores, particularly when multiple memory measures are administered. In turn, there is a substantial risk of psychometrically misclassifying MCI in healthy older adults. This study examined the base rates of low memory scores in older adults (55-87 years; n = 550) from the Wechsler Memory Scale-Third Edition (WMS-III; Wechsler, 1997b) standardization sample. The WMS-III consists of four co-normed episodic memory tests (i.e., Logical Memory, Faces, Verbal Paired Associates, and Family Pictures) that yield eight age- and demographically-adjusted standard scores (Auditory Recognition and Working Memory tests not included). When the eight age adjusted scores were examined simultaneously, 26% of older adults had one or more scores at or below the 5th percentile (i.e., -1.5 SDs). On the eight demographically- adjusted scores, 39% had at least one score at or below the 5th percentile. There was an inverse relationship between intellectual abilities and prevalence of low memory scores, particularly with the age-adjusted WMS-III scores. Understanding the base rates of low scores can reduce the overinterpretation of low memory scores and minimize false-positive misclassification. PMID- 18419846 TI - Functional implications of neuropsychological normality and symptom remission in older outpatients diagnosed with schizophrenia: A cross-sectional study. AB - Cognitive impairments in schizophrenia are well documented and correlated with functional disability. Although some patients demonstrate normal neuropsychological (NP) functioning, little is known about their functional disability. We examined the cross-sectional functional implications of NP normality and symptomatic remission in older outpatients diagnosed with schizophrenia or schizoaffective disorder, who were administered a NP battery and performance-based measures of functional and social competence, with their real world functioning rated by case managers. NP status was classified by the General Deficit Score (GDS) and remission status was based on the Positive and Negative Syndrome Scale (PANSS), yielding four subsamples of patients: NP normal-remitted (n = 21), NP normal-symptomatic (n = 22), NP impaired-remitted (n = 90), and NP impaired-symptomatic (n = 97). NP normal patients demonstrated better functional and social competence and better ratings of real world functioning, after controlling for premorbid abilities. However, compared to normative date, NP normal patients manifested disability in several real-world domains, including residential status. These results suggest that NP status is a better predictor of functional outcome then symptom status or the interaction of the two factors. The disability seen in NP normal cases indicates that factors other than cognitive impairments may determine aspects of everyday outcomes in schizophrenia. PMID- 18419847 TI - No disadvantage for the processing of global visual features in obsessive compulsive disorder. AB - The present study examined whether patients with obsessive-compulsive disorder (OCD) focus on details at the expense of global aspects. A recent study of our group using Navon letters (e.g., the letter "S" composed out of "A" letters) did not yield differences between OCD patients and controls on local processing. However, the task used may have lacked sensitivity, because it did not involve a response conflict condition (i.e., global and local level associated with different responses). In the current study, we gradually varied between-level conflict. Twenty-eight OCD patients and 30 healthy controls had to attend to the global and the local level of each item. OCD patients displayed comparable performance: patients neither displayed a preference to respond to the local level nor enhanced interference from the local level. In conclusion, the present study does not support the idea that a generalized bias to "miss the forest for the trees" forms part of the vulnerability to OCD. PMID- 18419848 TI - Influence of recruitment and participation bias in neuropsychological research among MS patients. AB - The potential influence of recruitment context in neuropsychological (NP) research is seldom addressed in the literature. Our experience with a previous large-scale study of MS cognitive impairment led us to speculate that referral questions and motivation bias significantly impact conclusions drawn from NP testing. We re-analyzed data from Benedict et al. (2006) and compared the results obtained across three groups of patients: paid research volunteers, clinical patients undergoing evaluation for routine monitoring of cognitive status, and clinical patients referred for diagnostic clarification, determination of disability benefits, and other more complex issues. Research volunteers were significantly younger and less frequently cognitively impaired compared to the clinical groups, greatly affecting prevalence estimates (45.6 to 65.6%). Significant correlations between depression and cognitive impairment were present only among the research volunteers. NP testing significantly predicted vocational outcomes in all groups. These results suggest that greater attention should be paid to the reasons patients agree to participate in NP testing. PMID- 18419849 TI - Comment on: 'antidepressant medications and other treatments of depressive disorders: a CINP Task Force report based on a review of evidence'. PMID- 18419850 TI - Comment on: 'antidepressant medications and other treatments of depressive disorders: a CINP Task Force report based on a review of evidence'. PMID- 18419853 TI - Use of host population reduction to control wildlife infection: rabbits and paratuberculosis. AB - Reduction in wildlife populations is a common method for the control of livestock infections which have wildlife hosts, but its success is dependent on the characteristics of the infection itself, as well as on the spatial and social structure of the wildlife host. Paratuberculosis (Mycobacterium avium subsp. paratuberculosis; Map) is a widespread and difficult infection to control in livestock populations and also has possible links to Crohn's disease in humans. Rabbits have recently been identified as a key wildlife species in terms of paratuberculosis persistence in the environment and risk to the wider host community, including cattle. Here we use a spatially explicit stochastic simulation model of Map dynamics in rabbit populations to quantify the effects of rabbit population control on infection persistence. The model parameters were estimated from empirical studies of rabbit population dynamics and rabbit-to rabbit routes of Map transmission. Three rabbit control strategies were compared: single unrepeated population reductions based on removing individual animals; single unrepeated population reductions based on removal of entire social groups; and repeated annual population reductions based on removing individual animals. Unrealistically high rabbit culls (>95% population reduction) are needed if infection is to be eradicated from local rabbit populations with a single one-off population reduction event, either of individuals or social groups. Repeated annual culls are more effective at reducing the prevalence of infection in rabbit populations and eradicating infection. However, annual population reductions of >40% are required over extended periods of time (many years). Thus, using an approach which is both highly conservative and parsimonious with respect to estimating lower bounds on the time to eradicate the infection, we find that Map is extremely persistent in rabbit populations and requires significant and prolonged effort to achieve control. PMID- 18419854 TI - Hospitalization rates for human metapneumovirus infection among 0- to 3-year-olds in Gipuzkoa (Basque Country), Spain. AB - Numerous studies have been published on human metapneumovirus (HMPV) infection, but few have been population based. The main aim of this study was to estimate the incidence rate of hospitalization for community-acquired HMPV infection in infants and children aged <3 years. Between July 2004 and June 2007, 796 episodes (742 patients) of community-acquired acute respiratory infection were hospitalized. HMPV was detected in 90 episodes (11.3%). Fifty-nine episodes occurred in infants aged <1 year. The mean length of hospital stay was 6.2 days (range 2-31 days). Thirteen children required admission to the intensive care unit. Viral co-infections were detected in 46 episodes (51.1%). The incidence rate of hospitalization per 1000 inhabitants was 2.6 (95% CI 2.1-3.2), lower than that for respiratory syncytial virus, but higher than that observed for the influenza and parainfluenza viruses. HMPV is a major respiratory pathogen that leads to a high hospitalization rate. PMID- 18419855 TI - Multiclonal outbreak of methicillin-resistant Staphylococcus aureus infections on a collegiate football team. AB - An outbreak of methicillin-resistant Staphylococcus aureus (MRSA) skin and soft tissue infections (SSTIs) occurred in a college football team in August 2006. Of 109 players on the team roster, 88 (81%) were interviewed during a cohort investigation. Twenty-five cases were identified, six of which were culture confirmed. Available culture isolates were typed by pulsed-field gel electrophoresis (PFGE), which identified two different MRSA strains associated with the outbreak. Playing positions with the most physical contact (offensive linemen, defensive linemen, and tight ends) had the greatest risk of infection [risk ratio (RR) 5.1, 95% confidence interval (CI) 2.3-11.5. Other risk factors included recent skin trauma (RR 1.9, 95% CI 0.95-3.7), use of therapeutic hydrocollator packs (RR 2.5, 95% CI 1.1-5.7), and miscellaneous training equipment use (RR 2.1, 95% CI 1.1-4.1). The outbreak was successfully controlled through team education and implementation of improved infection-control practices and hygiene policies. PMID- 18419856 TI - Applying an aggregative dispersive dichotomy (ADD) model to parasitic infections in host populations. AB - An aggregative dispersive dichotomy (ADD) model is presented to describe the distribution of parasites in host populations. The ADD model is a mathematical construct which provides two complementary measures extracted from a reformulated negative binomial (NBD) and an inequality model, which combine to capture observed patterns of a parasitic infection. The dispersion element is modelled using the NBD with the threshold set at a parasite level above zero. By applying binomial dichotomy, the host community is divided into two sub-populations, one including hosts harbouring parasites up to the threshold and the other with parasites above the threshold level. The k parameter, derived from the NBD, provides a cumulative probability. However, k is relatively insensitive to variations in the degree of aggregation, a known feature of the NBD model. The aggregation of parasites above the threshold in the host sub-population is evaluated by using an inequality model which is indexed by a scale-free parameter delta(delta >/= 1) and provides an accurate measure of parasite aggregation. Applications of this model are made from field and simulated data in wood mouse populations infected with the trichostrongylid nematode Heligmosomoides polygyrus from a woodland site in Surrey. PMID- 18419857 TI - Anthelmintic efficacy of Achillea millifolium against gastrointestinal nematodes of sheep: in vitro and in vivo studies. AB - Achillea millifolium Linn., commonly called 'Pehl-ghasa', is used by farmers in traditional animal health care as a vermifuge. The objective of this study was to evaluate the anthelmintic efficacy of crude aqueous extracts and crude ethanolic extracts of entire A. millifolium against the gastrointestinal nematodes of sheep. The worm motility inhibition assay was used for in vitro studies and faecal egg count reduction assay was used for in vivo studies. In vitro studies revealed significant anthelmintic effects of aqueous extracts and ethanolic extracts on live Haemonchus contortus worms (P < 0.05) as evident from their paralysis and/or death at 8 h post exposure. Aqueous extracts of A. millifolium resulted in a mean worm motility inhibition of 94.44%, while ethanolic extracts resulted in mean worm motility inhibition of 88.88%. The mean mortality index of aqueous extracts was 0.95 while for ethanolic extracts it was 0.9. The lethal concentration 50 was 0.05 mg ml(-1) for aqueous extracts and 0.11 mg ml(-1) for ethanolic extracts. The in vivo anthelmintic activity of aqueous and ethanolic extracts of A. millifolium demonstrated a maximum (88.40%) nematode egg count reduction in sheep treated with aqueous extracts at 2 g kg(-1) body weight on day 15 after treatment. Ethanolic extracts resulted in a maximum of 76.53% reduction in faecal egg counts on day 15 after treatment with 2 g kg(-1) body weight. Thus, the aqueous extracts exhibited greater anthelmintic activity under both in vitro and in vivo conditions, and could be due to the presence of water-soluble active principle/s in A. millifolium. It is concluded that the entire plant of A. millifolium possesses significant anthelmintic activity and could be a potential alternative for treating cases of helminth infections in ruminants. PMID- 18419858 TI - Differentiation between cholesteatoma and inflammatory process of the middle ear, based on contrast-enhanced computed tomography imaging. PMID- 18419876 TI - A comparison of levofloxacin and moxifloxacin use in hospitalized community acquired pneumonia (CAP) patients in the US: focus on length of stay. AB - OBJECTIVE: Length of stay (LOS) and hospitalization costs were compared among patients admitted for community-acquired pneumonia (CAP) and initially treated with either levofloxacin 750 mg intravenous (IV) or with moxifloxacin 400 mg IV. Hospital-related complications and relationship of LOS and comorbidities were descriptively examined. METHODS: A retrospective database study was conducted of adult patients admitted for CAP and given levofloxacin 750 mg IV or moxifloxacin 400 mg IV through the first 3 days of hospitalization, using the Premier Perspective comparative database. Cohorts were matched 1:1 by hospital geographic location, by coarse caliper propensity scores using all baseline covariates, and by Mahalanobis metric matching based on age and severity (All Patient Refined Diagnosis-related Groups Severity of Illness (APR-DRG SOI) index). Comparisons between groups were further adjusted for characteristics that remained imbalanced after matching using generalized estimating equation methodology. RESULTS: The initial sample of 3868 patients (levofloxacin = 827; moxifloxacin = 3041) was reduced to 1594 (797 patients per treatment group) after matching. Analyses of matched cohorts showed that the mean hospital LOS was significantly shorter for patients treated with levofloxacin 750 mg IV than for those patients treated with moxifloxacin 400 mg IV (5.8 vs. 6.4 days, respectively; least squares mean difference = 0.54 days; p = 0.020). Hospitalization costs were also lower for the levofloxacin 750 mg IV-treated patients (least squares mean difference = US$129; p = 0.753). There were no significant differences in the percentage of patients experiencing complications. LIMITATIONS: Although claims databases provide large sample sizes and reflect routine care, they do have several inherent limitations. Since randomization of subjects is not possible, adequate statistical techniques must be used to ensure treatment groups are balanced with respect to patient and clinical characteristics. In addition, data may be missing or miscoded. CONCLUSIONS: This retrospective study suggests that among patients hospitalized with CAP, initial treatment with levofloxacin 750 mg IV is associated with a significantly shorter mean hospital LOS compared with treatment with moxifloxacin 400 mg IV. The clinical implications of a shorter hospital LOS include improved patient and economic outcomes. PMID- 18419877 TI - Relative systemic availability of budesonide in patients with asthma after inhalation from two dry powder inhalers. AB - BACKGROUND: To improve dosing consistency and product features, budesonide inhalation powder delivered via a dry powder inhaler (DPI) (DPI-A 200 microg) was redesigned to include lactose, a newly shaped mouthpiece, and a new dose indicator (DPI-B). Budesonide DPI-B is available in two strengths (90 microg, 180 microg). OBJECTIVE: To compare the relative rate and extent of the systemic availability of budesonide inhaled via DPI-A and DPI-B and test for systemic absorption bioequivalence. METHODS: Adults (n = 37) with asthma as defined by the American Thoracic Society were randomized in an open-label, crossover, single center, single-dose study to budesonide DPI-A 200 microg x 4 inhalations, budesonide DPI-B 180 microg x 4 inhalations, or budesonide DPI-B 90 microg x 8 inhalations, on 3 days, each separated by a washout period of >or= 5 days. Plasma samples were collected immediately before and up to 12 h after dosing. Primary pharmacokinetic variables were area under the drug plasma concentration-time curve from 0 to infinity (AUC(0-infinity)) and maximum plasma concentration (C(max)); plasma concentration at 12 h (C(12h)) and time to maximum plasma concentration (T(max)) were secondary variables. Treatments were considered bioequivalent if the 90% confidence intervals (CIs) for their AUC(0-infinity) and C(max) ratios fell between 80 and 125%. Adverse events were collected. RESULTS: The 90% CIs for the ratios of AUC(0-infinity) and C(max) for budesonide DPI-A 200 microg and DPI-B 180 microg and for both budesonide DPI-B strengths fell between 80% and 125% (AUC(0-infinity): budesonide DPI-B 180 microg x 4/DPI-A 200 microg x 4: 96.3% [90% CI: 90.9, 102.1]; budesonide DPI-B 180 microg x 4/DPI-B 90 microg x 8: 92.2% [90% CI: 87.0, 97.7]; C(max): (budesonide DPI-B 180 microg x 4/DPI-A 200 microg x 4: 100.4% [95% CI: 92.1, 109.4]; budesonide DPI-B 180 microg x 4/DPI-B 90 microg x 8: 94.4% [90% CI: 86.6, 102.9]). No differences in C(12h) and T(max) were found between treatments. All treatments were well tolerated. CONCLUSIONS: Budesonide DPI-A 200 mug and DPI-B 180 mug have systemic absorption bioequivalence, and DPI-B 90 microg and 180 microg are dose-strength equivalent when administered at the same dose. These results may not be generalized to all patients with asthma, as this analysis included only patients with mild-to moderate asthma aged >or= 19 years. PMID- 18419878 TI - Evaluation of efficacy and safety of budesonide delivered via two dry powder inhalers. AB - BACKGROUND: The dry powder inhaler (DPI) device for budesonide inhalation powder 200 microg (DPI-A) was redesigned to improve dosing consistency and provide new features (budesonide inhalation powder 90 microg and 180 microg; DPI-B). OBJECTIVE: Two multicenter, parallel-group, double-blind, randomized, 12-week studies compared the efficacy and safety of budesonide delivered via each DPI versus placebo, and the systemic exposure of budesonide from each device. METHODS: Asthmatic adults with mild-to-moderate asthma (N = 621) and patients 6 17 years with mild asthma (N = 516) received budesonide DPI-B 360 microg or DPI-A 400 microg twice-daily (total daily dose 720 microg or 800 microg), budesonide DPI-B 180 microg or DPI-A 200 microg once daily (total daily dose 180 microg or 200 microg), or matching placebo. Change in forced expiratory volume in 1 second (FEV(1)) and secondary variables (asthma symptoms, beta(2)-adrenergic agonist use, peak expiratory flow [PEF], and withdrawals due to worsening asthma) versus placebo were measured. RESULTS: In both studies, FEV(1) significantly (p < 0.05) improved for all active treatments versus placebo except once-daily budesonide DPI-B 180 mug in adults. In the adult study, significantly (p < 0.05) greater improvements in all secondary variables occurred with all active treatments versus placebo. In the pediatric/adolescent study, improvements in AM/PM PEF were significantly (p or= 6 years with very mild asthma and adolescents and adults with mild-to-moderate asthma. The study is limited by the evaluation of only two doses for each product in both studies. Additionally, the studies were not designed to test equivalence or noninferiority between the active products. Pharmacokinetic characterization was limited because of the small sample sizes. PMID- 18419879 TI - An open-label, single-arm pilot study in patients with severe plaque-type psoriasis treated with an oral anti-inflammatory agent, apremilast. AB - OBJECTIVE: To evaluate the clinical and biological activity of apremilast in patients with severe plaque-type psoriasis. RESEARCH DESIGN AND METHODS: Apremilast, a phosphodiesterase-4 inhibitor, inhibits in vitro activity of multiple inflammatory factors implicated in the pathogenesis of psoriasis. Patients received 20 mg apremilast orally for 29 days. Immunohistological analysis was conducted on lesional-skin biopsies for psoriasis-associated inflammatory markers. Lipopolysaccharide-stimulated tumor necrosis factor-alpha levels were evaluated in blood. Psoriasis Area and Severity Index (PASI), static Physician's Global Assessment, and Body Surface Area were used to monitor disease severity. RESULTS: There were 19 patients enrolled in this study, of whom 17 completed the study. Epidermal thickness was reduced by a mean of 20.5% from baseline to day 29. Among the responders, T cells were reduced by 28.8% and 42.6% in the dermis and epidermis, respectively. Similarly, CD11c cells were reduced by 18.5% and 40.2% in the dermis and epidermis, respectively. Fourteen of the 19 (73.7%) patients demonstrated an improvement in their PASI scores. LIMITATIONS: This was a small, single-arm, open-label pilot study; therefore there was neither a placebo nor a comparison group. CONCLUSION: Apremilast demonstrated biological activity and improved psoriasis clinical efficacy scores in patients with severe plaque-type psoriasis. The majority of adverse events were mild in nature. Two adverse events (fatigue and dizziness) were judged by the investigator to be moderate and related to apremilast. In addition, there were no clinically relevant abnormal laboratory test results in subjects treated with apremilast for 29 days. PMID- 18419880 TI - Ethics and research: the case of informed consent. PMID- 18419881 TI - Lung structure and function in COPD. AB - The pathogenesis of chronic obstructive pulmonary disease (COPD) is related to a chronic innate and adaptive inflammatory immune response to inhaled toxic particles and gases, primarily as a result of the tobacco smoking habit. This inflammatory immune process develops in the lungs of everyone that smokes, and there is an association between the extent and severity of this tissue response and the severity of airflow limitation present in the fraction of the smoking population that develops COPD. This infiltration of inflammatory immune cells into the lung tissue is inextricably linked to a tissue repair and remodeling process that enlarges the bronchial mucus glands, thickens the walls and narrows the lumen of conducting airways <2 mm in diameter. A multivariate analysis has shown that thickening of the walls of the small conducting airways and occlusion of their lumen by inflammatory exudates containing mucus explains more of the variance in the association between FEV1 decline and histology in COPD than the infiltration of the tissue by any inflammatory cell type. Emphysematous destruction of the gas exchanging tissue also contributes to the airflow limitation by reducing the elastic recoil pressure available to drive air out of the lung during forced expiration. This tissue destruction begins in the respiratory bronchioles in very close proximity to the small conducting airways that become the major site of obstruction in COPD. The mechanism(s) that allow small airways to thicken in such close proximity to lung tissue undergoing emphysematous destruction remain a puzzle that needs to be solved. As the accumulation of tissue responsible for thickening the small conducting airways is a very different pathological process from the emphysematous destruction of surrounding gas exchanging tissue, we need a better understanding of the pathogenesis of both processes and better methods of separating their relative contribution to airflow limitation in individuals to adequately prevent and treat COPD. PMID- 18419882 TI - Risk factors for clustering of tuberculosis cases: a systematic review of population-based molecular epidemiology studies. AB - BACKGROUND: Many molecular epidemiology studies have been conducted to identify risk factors for clustering of tuberculosis (TB) cases in the population. OBJECTIVE: To estimate the impact of commonly investigated risk factors on TB clustering. METHODS: Ten electronic databases were searched up to January 2006 along with a hand search of the International Journal of Tuberculosis and Lung Disease and bibliographies of review articles. Meta-analyses of odds ratios (ORs) for various risk factors were conducted using random effect models, stratified by TB incidence. Meta-regressions were employed to account for the heterogeneity in clustering proportions and the magnitudes of risk. FINDINGS: The TB clustering proportion varied greatly (7.0-72.3%) among 36 studies in 17 countries. In multiple meta-regression analyses, high TB incidence, mean cluster size and conventional contact tracing were significantly associated with higher clustering. The pooled ORs (95%CIs) for low and high/intermediate TB incidence studies, using a cut off of 25/100000 per year, were 3.4 (2.7- 4.2) and 1.6 (1.3 2.1) for local-born status, 1.6 (1.5-1.7) and 1.7 (1.3-2.2) for pulmonary TB and 1.2 (1.1-1.3) and 1.3 (1.1-1.7) for smear-positive cases, respectively. Male sex, local birth, alcohol abuse and injection drug use were significantly higher risks in low TB incidence studies than in the high/intermediate ones. INTERPRETATION: Meta-analyses yielded significant estimates of ORs for several risk factors across both levels of TB incidence. Alcohol abuse, injection drug use and homelessness--all characteristics of marginalized populations--were found to be consistently significant in populations of low TB incidence. More research is needed to better understand TB transmission dynamics in high-burden countries. PMID- 18419883 TI - Starting treatment: caring for patients and their families. AB - The standards presented in this section focus on providing physical, social and psychological care for the patient at the point he or she is diagnosed with tuberculosis (TB) and starts treatment. Detailed guidance is included with regard to organising directly observed treatment (DOT) safely and acceptably for both the patient and the management unit. The aim is to give the patient the best possible chance of successfully completing treatment according to a regimen recommended by the World Health Organization. If the health service where the patient is diagnosed cannot offer ongoing treatment and care due to a lack of facilities, overcrowding or inaccessibility, the patient needs to be referred to a designated TB management unit (BMU) elsewhere. The patient may also receive treatment from a facility outside a BMU. However care is organised, it is essential for all patients who are diagnosed with TB to be registered at an appropriate BMU so that their progress can be routinely monitored and programme performance can be assessed. To avoid the risk of losing contact with the patient at any stage of their care, good communication is essential between all parties involved, from the patient him/herself to the person supervising their DOT to the BMU. PMID- 18419884 TI - Thinking in three dimensions: a web-based algorithm to aid the interpretation of tuberculin skin test results. AB - BACKGROUND: The tuberculin skin test (TST) is the most widely used test for detecting tuberculosis (TB) infection. Accurate interpretation of TST requires consideration of three dimensions-the size of the skin reaction, the positive predictive value (PPV) and risk of disease. METHODS: We developed a web-based algorithm incorporating epidemiological, medical and radiographic risk factors to help in the interpretation of positive TST results in adults (http://www.meakins.mcgill.ca/meakins/NEW TST Calculator/homeE.htm). We used summary estimates from published reviews on the prevalence of latent TB infection, the likelihood of false-positive TST and risk of active TB disease. RESULTS: The algorithm calculations show that the most important determinants of risk of active disease are the presence of medical and radiographic risk factors, while the size of the reaction is of modest importance. In persons who have received bacille Calmette-Guerin vaccination after infancy, the algorithm calculations show that the PPV will be low. In such persons, the risk of disease is predicted to be very low, unless there are medical or radiographic risk factors that increase the risk of reactivation. CONCLUSIONS: Our web-based algorithm can generate clinically useful estimates of the annual and cumulative lifetime risk of developing TB in adults with a positive TST. PMID- 18419885 TI - Tuberculosis in a low-incidence US area: local consequences of global disruptions. AB - SETTING: Tuberculosis (TB) in the United States is increasingly concentrated among foreign-born persons. The northeastern states, including Connecticut, are among those with the highest proportion of foreign-born patients. METHODS: This retrospective analysis of surveillance data from Connecticut for 1996 through 2005 compared TB case rates and risk factors among US-born and foreign-born persons. RESULTS: Between 1996 and 2005, TB cases declined by 8.7% in foreign born persons and by 53.6% in US-born persons. The median annual incidence rate for foreign-born persons was 19.7 cases per 100000 population compared with 1.5 for US-born persons. Refugees had the highest TB rate (116 cases/100000) in the first year of their arrival. Resistance to any drug was more common among foreign born persons (15.0%) than among US-born persons (9.3%). Although the proportion of multidrug-resistant TB was highest among foreign-born persons with prior TB (5.6%), most cases occurred in those without prior TB. Risk factors for TB, such as human immunodeficiency virus infection, drug use, incarceration and homelessness, were more common among US-born TB patients than among foreign-born patients. CONCLUSIONS: Although TB case rates for US-born persons in Connecticut have declined dramatically, foreign-born persons, including refugees fleeing conflict, contribute disproportionately to the TB burden. Future efforts to eliminate TB must be directed toward immigrants and refugees. PMID- 18419886 TI - Prevalence and risk factors for latent tuberculosis infection among health care workers in Georgia. AB - BACKGROUND: Tuberculosis (TB) is a major public health problem in Georgia, but few TB infection control measures have been implemented in health care facilities. OBJECTIVE: To assess the prevalence and risk factors for latent TB infection (LTBI) among Georgian health care workers (HCWs) using two diagnostic tests, the tuberculin skin test (TST) and the QuantiFERON-TB Gold In Tube test (QFT-3G), an interferon-gamma release assay. METHODS: A cross-sectional study was conducted between June and August 2006 among HCWs at the Georgian National TB Program. RESULTS: Of 265 HCWs enrolled, 177 (67%) had a positive TST and 159 (60%) had a positive QFT-3G; 203 (77%) had a positive result for at least one of the tests and 50% tested positive for both tests. There was moderately good agreement between the tests (74%, kappa = 0.43, 95%CI 0.33-0.55). In multivariate analysis, employment for >5 years was associated with increased risk of a positive TST (OR 5.09, 95%CI 2.77-9.33) and QFT-3G (OR 2.26, 95%CI 1.27-4.01); age >30 years was associated with an increased risk of a positive QFT-3G (OR 2.91, 95%CI 1.32-6.43). DISCUSSION: A high prevalence of LTBI was found among Georgian HCWs and longer duration of employment was associated with increased risk. These data highlight the need for effective TB infection control measures and provide important baseline information as TB infection control measures are implemented. PMID- 18419887 TI - Factors predictive of adherence to tuberculosis treatment, Valle del Cauca, Colombia. AB - BACKGROUND: Early diagnosis and treatment are fundamental to tuberculosis (TB) control. Nevertheless, the effectiveness of TB management continues to be influenced by treatment adherence. OBJECTIVE: To determine which factors are predictive of adherence to TB treatment at the time of diagnosis in Colombia. METHODS: A cohort of 300 patients newly diagnosed with TB was followed up over 26 weeks. Treatment adherence was measured by determining whether the patient took all or part of the 84 doses in the 26 weeks of treatment. A logistic analysis was carried out and the predictive power of the final variables was determined by means of a receiving operator curve analysis. RESULT: A high incidence of partial completion of treatment (65.6%) was found. Significant associated factors were 1) living away from the family, 2) overcrowding at home (> or =2 persons per bedroom), 3) lack of family support, 4) living >10 min away from the treatment facility and 5) not having used the services of the treatment facility before. CONCLUSION: Several factors can be measured on PTB diagnosis that would help identify those patients at higher risk for treatment non-adherence. The predictive value of each of these factors alone was weak, but if associated their predictive value was high. PMID- 18419888 TI - City tuberculosis control coordinators' perspectives of patient adherence to DOT in Sao Paulo State, Brazil, 2005. AB - SETTING: Thirty-six priority cities in Sao Paulo State, Brazil, with a high incidence of tuberculosis (TB) cases, deaths and treatment default. OBJECTIVE: To identify the perspectives of city TB control coordinators regarding the most important components of adherence strategies adopted by health care teams to ensure patient adherence in 36 priority cities in the State of Sao Paulo, Brazil. DESIGN: Qualitative research with semi-structured interviews conducted with the coordinators of the National TB Control Programme involved in the management of TB treatment services in the public sector. RESULTS: The main issues thought to influence adherence to directly observed treatment (DOT) by coordinators include incentives and benefits delivered to patients, patient-health care worker bonding and comprehensive care, the encouragement given by others to follow treatment (family, neighbours and health professionals), and help provided by health professionals for patients to recover their self-esteem. CONCLUSION: The main aspects mentioned by city TB control coordinators regarding patient adherence to treatment and to DOT in Sao Paulo are improvements in communications, relationships based on trust, a humane approach and including the patients in the decision-making process concerning their health. PMID- 18419889 TI - An ethnographic study of barriers to and enabling factors for tuberculosis treatment adherence in Timor Leste. AB - BACKGROUND: Tuberculosis (TB) is a major public health problem in Timor Leste; treatment adherence was identified by the National TB Control Programme (NTP) as an impediment to TB control. OBJECTIVE: To identify barriers to and enabling factors for the successful implementation of the DOTS strategy in Timor Leste. METHOD: Qualitative research was carried out in the two districts (one rural and one urban) with the lowest treatment completion rates. Semi-structured interviews and focus group discussions were conducted with patients, health workers and community members in eight villages. RESULT: Good knowledge of TB, including a correct understanding of how it is cured, together with the provision of incentives, were important factors contributing to treatment completion. Defaulting patients and community members had less knowledge of TB. TB nurses had a good understanding of, and a high level of commitment to implementing the DOTS strategy. Obstacles to treatment completion included preference for traditional medicine, economic difficulties and geographic remoteness. CONCLUSION: Local cultural practices and knowledge as well as socio-economic factors contribute to less than optimal adherence to TB treatment. This study has assisted Timor Leste's NTP in modifying its DOTS expansion strategies to overcome barriers to treatment completion. PMID- 18419890 TI - Decentralising tuberculosis services from county tuberculosis dispensaries to township hospitals in China: an intervention study. AB - SETTING: County tuberculosis (TB) dispensaries are the basic health care unit for the provision of TB services in China. In the rural hills of the western provinces of China, however, county centres are often far from patients' homes. OBJECTIVE: To evaluate whether decentralisation would result in improvements in patient access to TB services while maintaining the quality of the DOTS strategy. METHODS: The present study compared TB services that were decentralised to township hospitals with two comparable control groups where township hospitals continued to routinely refer suspects to the county TB dispensaries. Training and supervision of quality control were provided. Routine TB reporting data were reviewed. A questionnaire-based survey was conducted for 171 new TB patients enrolled from 1 April 2005 to 31 July 2006. RESULTS: Patients in the decentralised group spent less on travel and treatment for TB. The TB case notification rates increased significantly in the decentralised group, but remained unchanged in the control groups. Compared with the control groups, the decentralised group had improved treatment outcomes after 16 months. A higher quality of care was also observed in the decentralised group. CONCLUSION: The decentralised model showed better patient access and quality of care. The study demonstrated that decentralising TB services to the township level, especially vital in China's poorly accessible areas, was feasible. PMID- 18419891 TI - Molecular epidemiology of Mycobacterium tuberculosis in an urban area in Japan, 2002-2006. AB - SETTING: Shinjuku City, Tokyo, Japan. OBJECTIVE: To evaluate the status of transmission of Mycobacterium tuberculosis in Shinjuku City to allocate resources efficiently and effectively for a successful tuberculosis (TB) control programme. DESIGN: Observational descriptive study combining the genotype data of M. tuberculosis with TB patient profiles. RESULTS: The genotype clustering rate was significantly higher in males (adjusted odds ratio [aOR] 1.94, 95%CI 1.04-3.65, P = 0.038), patients aged <40 years (aOR 2.09, 95%CI 1.17-3.71, P = 0.012) and the homeless (aOR 2.72, 95%CI 1.42-5.20, P = 0.002), and was lower for the foreign born (aOR 0.21, 95%CI 0.06-0.76, P = 0.017). Among 45 genotype clusters containing 152 TB patients, 26 clusters containing 102 patients (67.1%) were composed of a mix of homeless and non-homeless patients. One of the mixed clusters included an 8-month-old infant born in Japan. CONCLUSION: The study revealed that M. tuberculosis transmission occurred more frequently among the homeless than in non-homeless persons. However, transmission by casual contact between the homeless and the general population was also shown to occur. PMID- 18419892 TI - Physician-initiated courtesy MODS testing for TB and MDR-TB diagnosis and patient management. AB - SETTING: Laboratorio de Investigacion de Enfermedades Infecciosas, Universidad Peruana Cayetano Heredia (UPCH) and government health centres, Lima, Peru. OBJECTIVE: To evaluate the contribution of unselected (courtesy) microscopic observation drug susceptibility (MODS) testing to the diagnosis and/or drug susceptibility testing (DST) of tuberculosis and their subsequent impact upon patient management. DESIGN: Retrospective database analysis and case note review of MODS culture-positive cases. RESULTS: Mycobacterium tuberculosis was isolated in 28.9% of 225 samples (209 patients); 22.2% of 63 positive cases were multidrug resistant. In 58 MODS culture-positive cases with follow-up data available, MODS provided culture confirmation of diagnosis, DST or both in 82.8%, before any standard method. In 41.4%, this result should have prompted a modification in patient management. Delays between laboratory result and initiation or change of treatment, where applicable, took on average 42 and 64 days, respectively, of which a delay of respectively 17 and 48 days occurred after the receipt of results by the health facility. CONCLUSION: MODS provides important data for clinical management within a meaningful timeframe and should contribute positively to patient outcomes due to earlier initiation of appropriate therapy. Although clinicians may successfully select patients likely to benefit from MODS, ongoing work is required to identify optimal implementation of the assay and to reduce logistical and health system derived delays. PMID- 18419893 TI - High levels of resistance to second-line anti-tuberculosis drugs among prisoners with pulmonary tuberculosis in Georgia. AB - SETTING: Penitentiary system of Georgia. OBJECTIVE: To determine the prevalence of resistance to second-line drugs among prisoners with pulmonary tuberculosis (PTB). DESIGN: Retrospective evaluation of resistance to second-line drugs in tuberculosis (TB) patients treated from 2001 to 2003. RESULTS: The overall observed prevalence of multidrug-resistant TB (MDR-TB) was 14.4% (39/270). The lowest resistance was found for ofloxacin (OFX), which was 2.2% (6/270) overall and 5.1% (2/39) among MDR patients. Isolates from four non-MDR patients who had never received anti-tuberculosis treatment were found to be resistant to OFX. Resistance to kanamycin and capreomycin occurred simultaneously only among MDR patients and was observed in 17/39 cases (43.6%). High rates of resistance to > or =2 second-line drugs (18/39, 46.2%) and > or =3 second-line drugs (10/39, 25.6%) were observed among all MDR-TB patients, reaching respectively 59.3% and 29.6% among previously treated MDR-TB cases. Only one patient was found to be resistant to four second-line drugs. No extensively drug-resistant TB (XDR-TB) according to the latest definition was detected. CONCLUSION: Our findings reveal a serious threat to the TB control efforts in the study population. PMID- 18419894 TI - Smoking behavior among former tuberculosis patients in Indonesia: intervention is needed. AB - SETTING: Five lung clinics in Jogjakarta Province, Indonesia. OBJECTIVE: To document smoking patterns among tuberculosis (TB) patients before diagnosis and following treatment, to identify smoking-related messages given by health professionals and DOTS providers and to identify predictors of smoking relapse. DESIGN: A cross-sectional survey of 239 male TB patients completed DOTS-based treatment during 2005-2006. Subjects were interviewed at home using a semi structured questionnaire. Female patients were excluded, as very few smoke. RESULTS: Most TB patients quit smoking when under treatment, but over one third relapsed at 6 months post-treatment. About 30% were never asked about their smoking behavior or advised about quitting. Of relapsed smokers, 60% received only general health messages and not TB-specific smoking messages. DOTS providers are not currently involved in cessation activities. The perception that any level of smoking is harmless for ex-TB patients was a significant predictor for smoking relapse. CONCLUSION: Physicians and DOTS providers should be actively involved in smoking cessation activities among TB and ex-TB patients. Based on these data, the Quit Tobacco Indonesia Project is mounting a pilot intervention to train DOTS providers, who are mostly family members of patients, to deliver smoking cessation messages and reinforce the cessation advice provided by physicians during and following TB treatment. PMID- 18419895 TI - Neonatal pulmonary tuberculosis evolving to a destroyed lung. AB - Tuberculosis (TB) in the newborn is infrequent, difficult to diagnose and often devastating. Congenital TB is rare, with most neonates and young infants becoming infected after birth. The incidence of neonatal TB might increase in industrialised countries as a result of immigration from countries with higher TB incidence among women of childbearing age. We report two cases of post-natally acquired pulmonary TB in newborns who developed marked lung destruction, a complication of TB which has seldom been described in the first month of life. A high index of clinical suspicion is required when evaluating pregnant women at risk for TB and their ill children, as early identification and treatment can prevent the devastating consequences of TB. PMID- 18419896 TI - Brazilian paediatricians' awareness of national guidelines for community-acquired pneumonia. AB - BACKGROUND: The Integrated Management of Childhood Illness (IMCI) strategy uses simple clinical signs for the diagnosis and severity evaluation of community acquired pneumonia (CAP). OBJECTIVE: To describe paediatrician awareness of the IMCI strategy for CAP. DESIGN: A cross-sectional study analysing a descriptive case of severe CAP, presented as part of the Brazilian Board of Paediatrician Evaluation (BBPE) tests. RESULTS: Eighty-six (774/898) per cent of paediatricians followed the IMCI protocol to treat CAP. Although hospitalisation was considered in 90% of the answers, only 35% based this decision on lower chest indrawing. CONCLUSION: The BBPE showed that most physicians are aware of the IMCI recommendations. PMID- 18419897 TI - Image processing techniques for identifying Mycobacterium tuberculosis in Ziehl Neelsen stains. AB - Worldwide, laboratory technicians tediously read sputum smears for tuberculosis (TB) diagnosis. We demonstrate proof of principle of an innovative computational algorithm that successfully recognizes Ziehl-Neelsen (ZN) stained acid-fast bacilli (AFB) in digital images. Automated, multi-stage, color-based Bayesian segmentation identified possible 'TB objects', removed artifacts by shape comparison and color-labeled objects as 'definite', 'possible' or 'non-TB', bypassing photomicrographic calibration. Superimposed AFB clusters, extreme stain variation and low depth of field were challenges. Our novel method facilitates electronic diagnosis of TB, permitting wider application in developing countries where fluorescent microscopy is currently inaccessible and unaffordable. We plan refinement and validation in the future. PMID- 18419898 TI - Climate factors associated with historic spruce beetle (Coleoptera: Curculionidae) outbreaks in Utah and Colorado. AB - This study investigated relationships between climate and historic spruce beetle, Dendroctonus rufipennis Kirby (Coleoptera: Curculionidae), outbreaks in northern and southeastern Utah and western Colorado between 1905 and 1996. A chronology of outbreak years was constructed from historic records, research papers, newspapers, and other sources of information. Historic climate data for the region included annual and mean monthly temperature and precipitation, in addition to Palmer drought severity index (PDSI) values estimated from tree rings. Classification and regression tree analysis (CART) was used to identify those climate factors most important for predicting historic spruce beetle outbreaks. The factors identified by the best CART model included mean December temperature, mean September temperature 1 yr before outbreak years, the mean estimated PDSI value of the 5-yr period before outbreak years, and mean October precipitation. The resulting model correctly classified nonoutbreak and outbreak years 67 and 70% of the time, respectively. PMID- 18419899 TI - A core set of microsatellite markers for western corn rootworm (Coleoptera: Chrysomelidae) population genetics studies. AB - Interest in the ecological and population genetics of the western corn rootworm, Diabrotica virgifera virgifera LeConte, has grown rapidly in the last few years in North America and Europe. This interest is a result of a number of converging issues related to the increasing difficulty in managing this pest and the need to characterize and understand gene flow in the context of insect resistance management. One of the key components needed for successful population genetics studies is the availability of suitable molecular markers. Using a standard group of microsatellite markers enables researchers from different laboratories to directly compare and share their data, reducing duplication of effort and facilitating collaborative work among laboratories. We screened 22 candidate microsatellite loci against five criteria to create a core set of microsatellite markers for D. v. virgifera population genetics studies. The criteria for inclusion were moderate to high polymorphism, unambiguous readability and repeatability, no evidence of null alleles, apparent selective neutrality, and no linkage between loci. Based on our results, we recommend six microsatellite markers to be included as a core set in future population genetics studies of D. v. virgifera along with any other microsatellite or genetic markers. As more microsatellites are developed, those meeting the criteria can be added to the core set. We encourage other groups of researchers with common interests in a particular insect species to develop their own core sets of markers for population genetics applications. PMID- 18419900 TI - Flight performance of the soybean aphid, Aphis glycines (Hemiptera: Aphididae) under different temperature and humidity regimens. AB - The soybean aphid, Aphis glycines (Matsumura), is native to eastern Asia and has recently invaded North America, where it is currently the most important insect pest of soybeans. The soybean aphid has spread rapidly within North America, presumably through a combination of active and passive (wind-aided) flight. Here, we studied the active flight potential of A. glycines under a range of environmental conditions using an aphid flight mill. Winged (alate) A. glycines were tested on a specially designed 32-channel, computer-monitored flight mill system. Aphids that were 12-24 h old exhibited the strongest flight behavior, with average flight durations of 3.3-4.1 h, which represented flight distances of 4.6-5.1 km. After the age of 72 h, A. glycines flight performance rapidly declined. The optimum temperature range for flight was 16-28 degrees C, whereas optimum relative humidity was 75%. Our findings show that A. glycines posseses a fairly strong active flight aptitude (ability and inclination) and point to the possibility of flight initiation under a broad range of environmental conditions. These results have the potential to aid forecasting and management protocols for A. glycines at the landscape level. PMID- 18419901 TI - Development of immature stages and comparative demography of two cucurbit attacking fruit flies in Reunion Island: Bactrocera cucurbitae and Dacus ciliatus (Diptera Tephritidae). AB - On Reunion Island, two species of Dacini, Bactrocera cucurbitae (Coquillett) and Dacus ciliatus Loew, infest 16 host plant species belonging to the family Cucurbitaceae from sea level to 1,600 m. These two species represent two primary pests of this plant family on the island. Melon fly, Bactrocera cucurbitae, and Ethiopian fruit fly, D. ciliatus, larval development was studied at four different constant temperatures (15, 20, 25, and 30 degrees C) with three host plants (cucumber, pumpkin, and squash). Adult life histories of these two species were studied at 25 degrees C with the three host plants. The results led to the conclusion that B. cucurbitae had a faster egg incubation time. Its preimaginal instars developed significantly faster than those of D. ciliatus independent of temperature. B. cucurbitae and D. ciliatus had similar mean preoviposition duration and egg hatching success. Fecundity was significantly higher for the melon fly on cucumber and pumpkin and lower on squash. Two distinctly different life- history patterns were evident: (1) later onset of reproduction, longer oviposition time, longer life span, and higher fecundity (B. cucurbitae) and (2) early reproduction, lower oviposition time, shorter life span, and lower fecundity (D. ciliatus). These results are useful for improving laboratory rearing methods and for building simulation models to predict Dacini population dynamics. PMID- 18419902 TI - Larval life history responses to food deprivation in three species of predatory lady beetles (Coleoptera: Coccinellidae). AB - We studied life history responses of larvae of three coccinellid species, Coleomegilla maculata (DeGeer), Hippodamia convergens Guerin-Meneville, and Harmonia axyridis (Pallas), when deprived of food for different periods of time during the fourth stadium. The coccinellid species did not differ in starvation resistance when larvae were starved throughout the stadium; however, for larvae fed only on day 1 of the stadium, H. convergens had the highest starvation resistance, followed by H. axyridis and then C. maculata. Percentage weight loss of larvae was affected by food deprivation period and coccinellid species. Both C. maculata and H. axyridis lost significantly more weight than H. convergens when starved throughout the fourth stadium. When deprived of food for 4 d of the stadium, C. maculata lost a higher percentage of initial body weight than H. axyridis. Percentage weight loss of H. convergens did not differ from that of C. maculata or H. axyridis. The weight of fourth instars and adults declined in an accelerating pattern as food deprivation period increased. However, food deprivation period had no significant effect on pupal development time for any of the three species or on larval development time for C. maculata and H. convergens. The increase in H. axyridis larval development time as a result of an increase in food deprivation period was curvilinear. Based on this laboratory study, it would seem that H. convergens is better able to cope with acute nutritional stress than either C. maculata or H. axyridis. PMID- 18419903 TI - Life history of the mealybug, Maconellicoccus hirsutus (Hemiptera: Pseudococcidae), at constant temperatures. AB - Important life history parameters of the mealybug, Maconellicoccus hirsutus (Green), were characterized on hibiscus (Hibiscus rosa-sinensis L.) cuttings at six constant temperatures between 15 and 35 degrees C. The development of M. hirsutus was the fastest at 27 degrees C, where the mealybugs completed development in approximately 29 d. The lower (T(min)) and upper (T(max)) developmental thresholds and the optimal developmental temperature (T(opt)) for the development of female mealybugs were estimated as 14.5, 35, and 29 degrees C, respectively. The thermal constant (K), which is the number of temperature-day or degree-day units required for development, of the females was 347 DD. The original distribution range prediction (based on T(min) = 17.5 degrees C and K = 300 DD) indicated that M. hirsutus could complete at least one generation in all of the continental United States. However, results of this study suggested that the distribution range of M. hirsutus may expand northward because of the lower T(min), and the predicted number of generations in a year may be lower because of the higher K required to complete each generation. The average cumulative survival rate of M. hirsutus at 25 and 27 degrees C was 72%, which was significantly higher than 51 and 62% at 20 and 30 degrees C, respectively. M. hirsutus reproduced sexually, with each mated female producing 260-300 eggs between 20 and 27 degrees C but only approximately 100 eggs at 30 degrees C. Female longevity was reduced from 28 d at 20 degrees C to 19-21 d at 25-30 degrees C. At 27 degrees C, the net reproductive rate (R(o)) was estimated at 165 female symbol/female symbol, the intrinsic rate of population increase (r(m)) was 0.119 (female symbol/female symbol/d), the generation time (T(G)) was 43 d, and the doubling time (DT) was 5.8 d. The life table statistics suggested that the currently released biological control agents, which have higher r(m) than M. hirsutus, will be able to complete more generations than the mealybug within the tested temperature range; thus, they are effective against M. hirsutus. PMID- 18419904 TI - Does dietary P affect feeding and performance in the mixed-feeding grasshopper (Acrididae) Melanoplus bivitattus? AB - Although consequences of limited dietary protein and carbohydrate to performance are well studied for terrestrial insect herbivores, the importance of phosphorus (P) remains poorly understood. We examined the significance of dietary P to performance in fifth-instar nymphs of the grasshopper Melanoplus bivittatus fed artificial diets. Consumption, digestion, developmental rate, and growth in response to different levels of P nested within standard-Protein and carbohydrate diets were determined. Developmental rate was slowest on high-P diets; protein:carbohydrate concentration and P in diets affected frass production and consumption. Approximate digestibility and conversion of digested food were primarily influenced by the protein:carbohydrate quality of the diet but not P. Mass gain was marginally lower in the low-Protein:high carbohydrate diet used in this study. At the individual level, other than small effects to developmental rate at high concentrations for M. bivittatus, dietary P otherwise seems to have little effect on nymphal performance. To the degree that it is important, effects of dietary P depend on the concentrations of protein and carbohydrate in the diet. PMID- 18419905 TI - Effect of seven different pollens on bio-ecological parameters of the predatory mite Typhlodromus foenilis (Acari: Phytoseiidae). AB - Development, immature survival, reproduction, and life table parameters of the predatory mite Typhlodromus foenilis Oudemans were evaluated on seven different plant pollens (almond, apple, pear, cherry, apricot, plum, and walnut). Experiments were conducted in environmental chambers at 20 +/- 1 degrees C with 65 +/- 5% RH and a photoperiod of 16:8 (L:D). T. foenilis successfully completed its development on all pollen diets. Female mean developmental time from egg to adult emergence ranged from 11.3 +/- 0.12 to 14.28 +/- 0.23 d, whereas that of males was from 10.81 +/- 0.17 to 14.17 +/- 0.17 d. The average life span of adult females varied from 41.0 +/- 1.46 to 65.45 +/- 3.89 d, and total fecundity ranged from 11.78 +/- 0.65 to 39.09 +/- 2.38 eggs/female. The intrinsic rate of increase of T. foenilis was highest on almond pollen (r(m) = 0.104/d) and was the shortest on walnut pollen (r(m) = 0.055/d). The results show that this predatory mite develops well on all pollens, with almond and apricot pollens being of high nutritional value. This study provides further information for mass rearing of T. foenilis, and these findings are discussed in relevance with the establishment of this species in the field. PMID- 18419906 TI - Developmental rate estimation and life table analysis for Halyomorpha halys (Hemiptera: Pentatomidae). AB - Egg and nymphal development were studied under constant temperatures for the newly introduced pest species, Halyomorpha halys (Stal) (Hemiptera: Pentatomidae). Development was assessed at seven constant temperatures (15-35 degrees C). Development to adult was completed at temperatures between 17 and 33 degrees C, with egg hatch also occurring at 15 degrees C. The relationship between temperature and developmental rate was evaluated using three developmental models. Of the models evaluated, the Briere-1 model was the best fit for the empirical data of egg and total development and for providing accurate values for the temperature threshold. Application of the linear degree day model estimated 537.63 DD are needed for total development (egg to imaginal ecdysis). An additional 147.65 DD are needed for the preoviposition period of the female. Reproductive parameters were evaluated at 25 degrees C and indicate a median number of 28 eggs per egg mass. Oviposition occurred at 4.32-d intervals, and a female can continue to oviposit throughout its lifespan. H. halys is univoltine in New Jersey and Pennsylvania, but if it spreads to warmer climates in the United States, it could have multiple generations per year. PMID- 18419907 TI - Development of a host-based semiochemical lure for trapping emerald ash borer Agrilus planipennis (Coleoptera: Buprestidae). AB - Bark volatiles from green ash Fraxinus pennsylvanica were tested for electrophysiological activity by Agrilus planipennis using gas chromatographic electroantennographic detection (GC-EAD) and for behavioral activity using baited purple traps in Michigan. GC-EAD analysis of the headspace volatiles of bark tissue samples from 0- and 24-h-old fully girdled (stressed) ash trees showed that the latter had elevated sesquiterpene levels. Six of the elevated compounds consistently elicited antennal responses by both male and female A. planipennis. Five of the antennally active compounds were identified as alpha-cubebene, alpha copaene, 7-epi-sesquithujene, trans-beta-caryophyllene, and alpha-humulene (alpha caryophyllene). The sixth EAD-active compound remains unidentified. We monitored capture of adult A. planipennis on traps baited with several combinations of ash tree volatiles. Treatments included two natural oil distillates (Manuka and Phoebe oil) that were found to contain, respectively, high concentrations of four and five of the six antennally active ash bark volatiles. A four-component leaf lure developed by the USDA Forest Service and Canadian Forest Service was also tested. In three separate field studies, Manuka oil-baited traps caught significantly more adult beetles than unbaited traps. Lures designed to release 5, 50, and 500 mg of Manuka oil per day all caught more insects than unbaited traps. In a field test comparing and combining Phoebe oil with Manuka oil, Phoebe oil-baited traps caught significantly more beetles than either Manuka oil-baited traps or unbaited traps. We hypothesize that the improved attractancy of Phoebe oil to A. planipennis over Manuka oil is caused by the presence of the antennally active sesquiterpene, 7-epi-sesquithujene. PMID- 18419908 TI - Relative attractiveness of incomplete and complete blends of synthetic pheromone to male obliquebanded leafroller (Lepidoptera: Tortricidae) moths in a flight tunnel and in apple orchards: implications for sex pheromone-mediated mating disruption of this species. AB - The relative attractiveness of synthetic Choristoneura rosaceana (Harris) pheromone consisting of the major compound Z-11-tetradecenyl acetate (Z11 14:OAc), or the major compound plus one, two, or all three of the minor compounds E-11-tetradecenyl acetate (E11-14:OAc), Z-11-tetradecenol (Z11-14:OH), and Z-11 tetradecenal (Z11-14:Ald), in amounts similar to those in the natural pheromone, was compared in a flight tunnel and in apple orchards. In the flight tunnel, there was an increase in the proportion of moths that were activated when E11 14:OAc was combined with the main compound, but no detectible additional increases in response with the addition of Z11-14:OH and Z11-14:Ald. The time required for activation was twice as long when using the major compound than when using the major compound and one or more of the minor compounds. There was a trend of increase in the proportion of males initiating the take-off, lock-on, close-in, and touchdown phases of upwind flight when progressively more complete blends were used. The time required for the initiation of these behavioral phases was statistically similar for each treatment. The proportion of moths landing at the pheromone source was 10 times greater when using the complete blend than when using the main compound alone. Similar results were obtained in apple orchards, where the four-compound blend was 17-55 times more attractive than the main compound. The superior attractiveness of the four-compound pheromone compared with the major compound, or to the major compound plus the minor compound E11 14:OAc, suggests that it would be the most effective synthetic pheromone if competitive attraction were an important mechanism of disruption in this species. PMID- 18419909 TI - Reproduction-flight relationship in the beet armyworm, Spodoptera exigua (Lepidoptera: Noctuidae). AB - To evaluate the relationship between reproduction and migratory flight, we examined ovaries, tethered flight, and egg laying in the beet armyworm, Spodoptera exigua (Hubner). Ovarian development in female S. exigua was classified into five stages in relation to the postemergence age of the moths. Oviposition peaked at ovarian stages III and IV. Mating accelerated ovarian development in this species, and thus the mated females developed faster than the virgin moths in ovaries. In virgin adults, flight capacity increased from the first day to the third day after emergence and remained at a high level until the moths were 7 d old. In contrast, the mated moths reached the maximum flight capacity in 2 d after emergence and maintained a high level until the age of 7 d. However, mating status could not influence the flight capacity of the beet armyworm, although there were general differences in flight capacity between virgin and mated adults at the same age. Moreover, significant correlations between the number of eggs laid and the flight capacity were not performed by the mated moths. These results showed that mating status and ovarian development did not seem to influence flight capacity in the beet armyworm, which was not in agreement with the definitions of the oogenesis-flight syndrome observed in many other migratory insects. PMID- 18419910 TI - Spatial analysis of Lobesia botrana (Lepidoptera: Tortricidae) male population in a mediterranean agricultural landscape in central Italy. AB - The results obtained from the spatial analysis of pheromone-baited trap catch data of Lobesia botrana (Denis and Schiffermuller) males are reported. The research was undertaken in the Abruzzo region of central Italy. In the study area, vineyards (of Vitis vinifera L.) are the predominant cultivation, surrounded by hedgerows and small woodlots, and interspersed with cereal crops and olive groves. The main purpose of the study was to investigate the spatio temporal dynamics of L. botrana, inside and outside vineyards, and to evaluate the effect of the landscape elements on pest distribution. A trend orientation over the experimental area was observed along the direction from northwest to southeast. Correlograms fitted using a spherical model showed in all cases an aggregated distribution and an estimated range having a mean of 174 m in 2005 and 116 m in 2006. Contour maps highlighted that spatial distribution of L. botrana was not limited to vineyards, but its presence is high particularly inside olive groves. The adult distribution on the experimental area changed during the season: hot spots of flight I were positioned inside olive groves; during flights II and III, they were concentrated in vineyards. L. botrana males were also captured in uncultivated fields, but never in high densities. Our results showed that a large proportion of the adult population of L. botrana inhabits areas outside those usually targeted by pest management programs. Thus, in Mediterranean agro-ecosystems, it is highly recommended to consider the whole landscape, with particular attention to olive crops. PMID- 18419911 TI - Yellow sticky trap catches of parasitoids of Bemisia tabaci (Hemiptera: Aleyrodidae) in vegetable crops and their relationship to in-field populations. AB - We examined the relationship of yellow sticky trap captures of Bemisia tabaci (Gennadius) biotype B parasitoids to the local population of parasitoids as measured by leaf samples of parasitized whiteflies and mass release of parasitoids. Traps were placed in experimental collard and cowpea field plots in Charleston, SC, and in commercial organic fields of spring cantaloupe and watermelon in the Imperial Valley, CA. The exotic parasitoid Eretmocerus emiratus Zolnerowich and Rose was released in Imperial Valley fields to ensure parasitoid populations would be present. Bemisia adults were trapped in the greatest numbers on the upper surface of horizontally oriented sticky traps in melon fields. In contrast, the lower trap surfaces consistently captured more Eretmocerus than upper surfaces. Female parasitoids were trapped in greater numbers than males, especially on the lower trap surfaces. Progeny of released exotic Eretmocerus greatly outnumbered native E. eremicus Rose and Zolnerowich and Encarsia spp. on traps. Throughout the season, the trend of increasing numbers of Eretmocerus on traps parallelled the increase in numbers of whiteflies. Over the season, 23-84% of all B. tabaci fourth instars were visibly parasitized by Eretmocerus. The numbers of Eretmocerus caught by traps in cantaloupe were similar in trend to numbers on leaf samples in melons, but not with those in watermelon, where whitefly populations were lower. Parasitoid numbers were low in collard and cowpea samples, and no trend was observed in numbers of parasitoids captured on traps and numbers on leaves for these two crops. Overall, there were no significant correlations between sticky trap catches of parasitoids and numbers of parasitized whiteflies on leaf samples in any test fields. Nevertheless, sticky traps placed within crops may be useful for observing trends in whitefly parasitoid populations at a particular site and for detecting parasitoids at specific locations. PMID- 18419912 TI - Worker size in the formosan subterranean termite in relation to colony breeding structure as inferred from molecular markers. AB - The Formosan subterranean termite, Coptotermes formosanus Shiraki, is an invasive species that originated in China and has been introduced to Hawaii and the U.S. mainland. Colonies are headed either by a pair of reproductives (simple families) or by varying numbers of inbreeding reproductives (extended families), and therefore have variable degrees of inbreeding. Worker size also varies among colonies of Formosan termites. We tested whether variation in worker size can be explained by the breeding system. Workers were collected from colonies from three geographically separated populations (China, Hawaii, and Louisiana), and body weight and head size were measured. Microsatellite genotyping was used to establish whether colonies were simple or extended families and to determine the heterozygosity of workers and their degree of inbreeding relative to their colony (F (IC), sensitive to the number of reproductives). All Chinese colonies contained multiple inbreeding neotenics. In Hawaii, 37% of the colonies were simple families and 63% were extended families, both having considerable degrees of inbreeding. In Louisiana, 57% of the colonies were simple families, which were mostly headed by unrelated pairs, and 43% were extended families. In simple families, size and body weight of workers were not associated with F (IC) or heterozygosity. In extended families of two populations, both size parameters were negatively correlated with F (IC); however, heterozygosity was not associated with worker size in any of the populations. This suggests that the number of reproductives within colonies has a stronger influence on worker size than the individuals' genetic diversity in Formosan subterranean termite colonies. PMID- 18419913 TI - Spatio-temporal ecology and management of cabbage maggot. AB - This study analyzes the spatio-temporal dispersion patterns of the cabbage maggot (Delia radicum L.) (Diptera: Anthomyiidae) infestation in rutabagas and turnips in Oregon and suggests ways to exploit the spatial and temporal ecology of the cabbage maggot to improve management of the pest. The patchy distribution of cabbage maggots arises from a combination of first-order effects driven by spatial heterogeneity and second-order effects driven by spatial autocorrelation. The intensity of cabbage maggot infestations varied from year to year. Within a given year, damage rates tended to be higher in rutabagas than turnips, in crops planted earlier in the season, and in fields near nurseries and houses. Nonsignificant first-order effects included soil texture, distance from river, proximity to maggot sources (other than cultivated fields), type of vegetation on field borders, field manager, field area, and perimeter. Second-order effects were processes intrinsic to the population and would give rise to patchiness even in a homogeneous environment. For example, adults may be attracted to others of their species or eggs may be deposited in batches. The locations of patches arising from second-order effects cannot be predicted from knowledge of environmental covariates. However, cabbage maggot does not tend to disperse far, and existing patches tend to give rise to other patches nearby at a later time. We found elevated damage rates in spring fields planted near fields that were heavily damaged the previous fall and in fields planted late in the season near fields that had heavy damage early in the season. PMID- 18419914 TI - Do spider diversity and assemblages change in different contiguous habitats? A case study in the protected habitats of the humid chaco ecoregion, northeast Argentina. AB - Spiders are a megadiverse group that can be useful indicators of the overall species richness and health of biotic communities. The spider diversity in subtropical forests of the Neotropical region are not yet well known, especially in Argentinean subtropical forests where systematic fieldwork has not been done until recently. The Great Chaco is very important as the unique dry subtropical forest of the earth, but it is suffering increasing degradation by the advance of agriculture. Spider communities have been shown to be more directly influenced by vegetation architecture than vegetation species composition. In this study, we aim to assess whether spider diversity and assemblages change in adjacent habitats with different types of vegetation. We compare the diversity and spider assemblages in two different contiguous protected habitats (hygrophilous woodland and savannah parkland) of the Mburucuya National Park, (Humid Chaco ecoregion). Seasonal samples were obtained using three types of sampling methods: pitfall trapping, beating, and manual litter extraction. The spider assemblages were different in the studied areas, and the abundance, diversity, evenness, and species richness were higher in the hygrophilous woodland than the savannah parkland. These differences in spider diversity and assemblages indicate that both types of habitats are important if the biodiversity is to be conserved in the Chaco ecoregion, where different types of habitat are shown as a patchy distribution. PMID- 18419915 TI - Trophic associations of a dung beetle assemblage (Scarabaeidae: Scarabaeinae) in a woodland savanna of Botswana. AB - Species richness and abundance of dung beetles were assessed across a range of bait types that acted as surrogates for the food resources available in Chobe National Park, Botswana. These bait types were comprised of the dung of pig (omnivore), cattle (ruminant herbivore dropping fine-fiberd pads), sheep (pellet dropping ruminant herbivore), and elephant (monogastric, nonruminant herbivore producing coarse-fibered droppings), and chicken livers (carrion). Species richness was similar between traps baited with pig, cattle, and elephant dung but was relatively lower in those baited with sheep dung and carrion. In traps baited with pig dung, abundance was relatively greater than in all other bait types. A cluster analysis of species abundance distributions for the 30 most abundant species identified four different patterns of bait type association at a 60% level of similarity. All but 1 of the 15 species in cluster A were attracted primarily to the dung of omnivores and pad-dropping ruminant herbivores (pig and cattle). All seven species of cluster B were attracted primarily to coarse fibered, nonruminant herbivore dung (elephant). All four species of cluster C were primarily carrion and pig dung associated, whereas all four species of cluster D were carrion specialists. In conclusion, the most abundant species were attracted to all bait types, but most species were largely specialized to different dung types or carrion, with dung attracting the majority of the fauna in terms of both species richness and abundance. PMID- 18419916 TI - Spatio-temporal analysis of Xyleborus glabratus (Coleoptera: Curculionidae [corrected] Scolytinae) invasion in eastern U.S. forests. AB - The non-native redbay ambrosia beetle, Xyleborus glabratus Eichhoff (Coleoptera: Curculionidae: Scolytinae), has recently emerged as a significant pest of southeastern U.S. coastal forests. Specifically, a fungal symbiont (Raffaelea sp.) of X. glabratus has caused mortality of redbay (Persea borbonia) and sassafras (Sassafras albidum) trees in the region; several other Lauraceae species also seem susceptible. Although the range of X. glabratus continues to expand rapidly, little is known about the species' biology and behavior. In turn, there has been no broad-scale assessment of the threat it poses to eastern U.S. forests. To provide a basic information framework, we performed analyses exploiting relevant spatio-temporal data available for X. glabratus. First, we mapped the densities of redbay and sassafras from forest inventory data. Second, we used climate matching to delineate potential geographic limits for X. glabratus. Third, we used county infestation data to estimate the rate of spread and modeled spread through time, incorporating host density as a weighting factor. Our results suggest that (1) key areas with high concentrations of redbay have yet to be invaded, but some are immediately threatened; (2) climatic conditions may serve to constrain X. glabratus to the southeastern U.S. coastal region; and (3) if unchecked, X. glabratus may spread throughout the range of redbay in <40 yr. Disruption of anthropogenic, long-distance dispersal could reduce the likelihood of this outcome. PMID- 18419917 TI - Habitat associations of ant species (Hymenoptera: Formicidae) in a heterogeneous Mississippi landscape. AB - Community assembly and the factors that influence it have long been a topic of interest to ecologists, but theory has yet to produce unequivocal evidence that communities assemble in predictable ways. The goal of this study was to document the relationship between ant communities and environmental variation between four habitat types. To accomplish this, ant communities and 16 environmental variables were sampled across four different habitat types in the Black Belt Prairie and Flatwoods regions in Mississippi. Furthermore, ant species were placed into functional groups for an analysis of the relationship between the assembly of ecological communities and variation in ecosystem function. A total of 20,916 ants representing 68 species was collected across the four habitat types. Nonmetric multidimensional scaling and analysis of covariance analyses both revealed three distinct ant communities, which can be characterized by habitat type: pasture, prairie, and woodland. Principle components analysis (PCA) simplified the 16 environmental variables into four principle components that explained 78% of the variation among sites. Results of multiple regression using the four PCA axes as predictor variables suggest that regional variation in soil structure, land cover type, and the presence of grazing have had major influences on ant community composition. Variation in flora and habitat architecture had smaller but significant effects on ant species diversity and functional group composition. Our results imply that restoration of native ant communities in disturbed habitats must consider how current disturbance regimens likely interact with the presence of Solenopsis to lower ant biodiversity. PMID- 18419918 TI - Oribatid mite communities in the canopy of montane Abies amabilis and Tsuga heterophylla trees on Vancouver Island, British Columbia. AB - To study the oribatid mite community inhabiting microhabitats in the canopy of montane Abies amabilis [(Douglas ex D. Don) Lindl.] and Tsuga heterophylla [(Raf.) Sarg] tree species across five elevational sites, we collected 180 branch tips and 180 foliose/crustose lichen samples over three time periods. Thirty three species of oribatid mites were identified from the study area. Mite species richness and abundance was significantly affected by microhabitat, and this association was independent of sampling time. At the microhabitat scale, distinct species assemblages were associated with lichen and branch tip habitats, and to a lesser degree, tree species. Conifer specificity was most apparent in the closely related species of Jugatala, where Jugatala tuberosa Ewing was only found on branch tips from A. amabilis and Jugatala sp. was primarily found on branch tips from T. heterophylla. Microhabitat specificity was most pronounced in Dendrozetes sp. where most individuals were found on branch tips and Anachiperia geminus Lindo et al. that occurred primarily on lichens. Principal components analysis of oribatid mite community composition further showed a high degree of association with microhabitat and tree species. Habitat profiles are difficult to discern for many species because tree, microhabitat, and elevation preferences confound distribution patterns. Given the significant tree-microhabitat associations in species composition in this montane canopy study, we suggest that sampling multiple microhabitats across elevations to look for patterns in community structure offers opportunities to explicitly test organizing principles in community ecology. PMID- 18419919 TI - Primary and secondary parasitoids (Hymenoptera) of aphids (Hemiptera: Aphididae) on blueberry and other Vaccinium in the Pacific Northwest. AB - Blueberry scorch virus, a commercially important Carlavirus in highbush blueberry, Vaccinium corymbosum L., is vectored by aphids (Hemiptera: Aphididae). We surveyed the aphids, primary parasitoids (Hymenoptera: Aphelinidae, Braconidae), and associated secondary parasitoids (Hymenoptera: Charipidae, Megaspilidae, Pteromalidae) on highbush blueberry and other Vaccinium in the Pacific Northwest from 1995 to 2006, with samples concentrated in 2005 and 2006, to lay the groundwork for augmentative biological control. Ericaphis fimbriata (Richards) was the principal aphid. The dominant parasitoid species were Praon unicum Smith, Aphidius n. sp., A. sp., and Aphidius ervi Haliday. Their frequency in relation to the other primary parasitoids varied significantly with geographical area; P. unicum dominated the frequency distribution in southwestern British Columbia, A. n. sp., west of the Cascades, and A. sp. and A. ervi east of the Cascades. Among the secondary parasitoids, pteromalids dominated, and their frequency in relation to the other secondary parasitoids was lowest in southwestern British Columbia. The parasitization rate for P. unicum and A. n. sp. in southwestern British Columbia increased from May or June to a maximum of 0.080 +/- 0.024 and 0.090 +/- 0.084 (SD), respectively, in late July or early August. P. unicum emerged in the spring 4 wk before A. n. sp. The parasitization rate for P. unicum was lower in conventional than organic fields. Whereas aphid density increased monotonically, P. unicum had two spring peaks. A simulation model showed that these peaks could reflect discrete generations. Releases of insectary-reared P. unicum at 150 or 450 DD above 5.6 degrees C, summing from 1 January, may effectively augment the natural spring populations by creating overlapping generations. PMID- 18419920 TI - Spatial and temporal variation in natural enemy assemblages on Maryland native plant species. AB - Habitat manipulation is a branch of conservation biological control in which vegetation complexity and diversity are increased in managed landscapes to provide food and other resources for arthropod natural enemies. This is often achieved by maintaining noncrop plant material such as flowering strips and beetle banks that provide natural enemies with nectar and pollen, alternative prey, shelter from disturbance, and overwintering sites. In most cases, plant material used in habitat manipulation programs is not native to the area in which it is planted. Using native plant species in conservation biological control could serve a dual function of suppressing pest arthropod outbreaks and promoting other valuable ecosystem services associated with native plant communities. We evaluated 10 plant species native to Maryland for their attractiveness to foliar and ground-dwelling natural enemies. Plants that showed particular promise were Monarda punctata, Pycnanthemum tenuifolium, and Eupatorium hyssopifolium, which generally harbored the greatest abundance of foliar predators and parasitoids, although abundance varied over time. Among ground-dwelling natural enemies, total predator and parasitoid abundance differed between plant species, but carabid and spider abundance did not. Matching certain plant species and their allied natural enemies with specific pest complexes may be enhanced by identifying the composition of natural enemy assemblages at different times of year and in both foliar and ground habitat strata. PMID- 18419921 TI - Plant resources and colony growth in an invasive ant: the importance of honeydew producing Hemiptera in carbohydrate transfer across trophic levels. AB - Studies have suggested that plant-based nutritional resources are important in promoting high densities of omnivorous and invasive ants, but there have been no direct tests of the effects of these resources on colony productivity. We conducted an experiment designed to determine the relative importance of plants and honeydew-producing insects feeding on plants to the growth of colonies of the invasive ant Solenopsis invicta (Buren). We found that colonies of S. invicta grew substantially when they only had access to unlimited insect prey; however, colonies that also had access to plants colonized by honeydew-producing Hemiptera grew significantly and substantially ( approximately 50%) larger. Our experiment also showed that S. invicta was unable to acquire significant nutritional resources directly from the Hemiptera host plant but acquired them indirectly from honeydew. Honeydew alone is unlikely to be sufficient for colony growth, however, and both carbohydrates abundant in plants and proteins abundant in animals are likely to be necessary for optimal growth. Our experiment provides important insight into the effects of a common tritrophic interaction among an invasive mealybug, Antonina graminis (Maskell), an invasive host grass, Cynodon dactylon L. Pers., and S. invicta in the southeastern United States, suggesting that interactions among these species can be important in promoting extremely high population densities of S. invicta. PMID- 18419922 TI - Predators associated with the hemlock woolly adelgid (Hemiptera: Adelgidae) in the Pacific Northwest. AB - The hemlock woolly adelgid, Adelges tsugae Annand (Hemiptera: Adelgidae), is causing widespread mortality of eastern hemlock, Tsuga canadensis L. Carriere, in the eastern United States. In western North America, feeding by A. tsugae results in negligible damage to western hemlock, Tsuga heterophylla (Raf.) Sargent. Host tolerance and presence of endemic predators may be contributing to the relatively low levels of injury to T. heterophylla caused by A. tsugae. Field surveys of the predator community associated with A. tsugae infestations on 116 T. heterophylla at 16 sites in Oregon and Washington were conducted every 4-6 wk from March 2005 through November 2006. Fourteen uninfested T. heterophylla were also surveyed across 5 of the 16 sites. Each sample tree was assigned an A. tsugae population score ranging from 0 to 3. Predators collected from A. tsugae-infested T. heterophylla represent 55 species in 14 families, listed in order of abundance: Derodontidae, Chamaemyiidae, Hemerobiidae, Coccinellidae, Cantharidae, Reduviidae, Miridae, Syrphidae, Chrysopidae, Coniopterygidae, Staphylinidae, Anthocoridae, Nabidae, and Raphidiidae. Laricobius nigrinus Fender (Coleoptera: Derodontidae), Leucopis argenticollis Zetterstedt (Diptera: Chamaemyiidae), and Leucopis atrifacies (Aldrich) (Chamaemyiidae) were the most abundant predators; together comprising 59% of predator specimens recovered. Relationships among predators and A. tsugae were determined through community structure analysis. The abundances of Laricobius spp. larvae, L. nigrinus adults, Leucopis spp. larvae, and L. argenticollis adults were found to be positively correlated to A. tsugae population score. Predators were most abundant when the two generations of A. tsugae eggs were present. L. argenticollis and L. atrifacies were reared on A. tsugae in the laboratory, and host records show them to feed exclusively on Adelgidae. PMID- 18419923 TI - Immature house fly (Musca domestica) control in breeding sites with a new Brevibacillus laterosporus formulation. AB - A bacterial formulation containing spores of a Brevibacillus laterosporus strain from Sardinia, known to be toxic by ingestion to the house fly (Musca domestica), was assayed in laboratory, outdoor, and field conditions for the control of immature stages of this pest. In all laboratory assays, the bacterial formulation exhibited toxicity against house fly larvae. A concentration of 1 x 10(8) spores of B. laterosporus/g of diet caused 100% mortality of house fly immature stages. The same formulation, applied at a concentration of 1 x 10(8) spores/ml, equivalent to a dose of 2 liters/m(2), caused a reduction in adult emergence from laboratory and natural breeding substrates (outdoor cage experiments) up to 80.3 and 57.8%, respectively. Similarly, this formulation applied in the cow pen of a diary farm at a dose of 2 liters/m(2) produced a significant reduction (30%) in immature fly development. Therefore, the use of this bacterial preparation in microbiological control strategies for the integrated pest management of this species is promising. PMID- 18419924 TI - Influence of seed head-attacking biological control agents on spotted knapweed reproductive potential in western Montana over a 30-year period. AB - Five insect biological control agents that attack flower heads of spotted knapweed, Centaurea stoebe L. subsp. micranthos (Gugler) Hayek, became established in western Montana between 1973 and 1992. In a controlled field experiment in 2006, seed-head insects reduced spotted knapweed seed production per seed head by 84.4%. The seed production at two sites in western Montana where these biological control agents were well established was 91.6-93.8% lower in 2004-2005 than 1974-1975, whereas the number of seed heads per square meter was 70.7% lower, and the reproductive potential (seeds/m(2)) was 95.9-99.0% lower. The average seed bank in 2005 at four sites containing robust spotted knapweed populations was 281 seeds/m(2) compared with 19 seeds/m(2) at four sites where knapweed density has declined. Seed bank densities were much higher at sites in central Montana (4,218 seeds/m(2)), where the insects have been established for a shorter period. Urophora affinis Frauenfeld was the most abundant species at eight study sites, infesting 66.7% of the seed heads, followed by a 47.3% infestation by Larinus minutus Gyllenhal and L. obtusus Gyllenhal. From 1974 to 1985, Urophora spp. apparently reduced the number of seeds per seed head by 34.5 46.9%; the addition of Larinus spp. further reduced seed numbers 84.2-90.5% by 2005. Path analysis indicated that both Larinus spp. and U. affinis contributed significantly to reduction of seed production over the 30-yr period. Spotted knapweed density may not decrease significantly until the seed bank falls below a critical threshold. PMID- 18419925 TI - Ovipositional biology of Viburnum leaf beetle, Pyrrhalta viburni (Coleoptera: Chrysomelidae). AB - Pyrrhalta viburni Paykull, a new landscape pest in the United States, feeds in both the larval and adult stages on foliage of plants in the genus Viburnum. We measured lifetime oviposition capacity of mated and unmated females reared in the laboratory versus field-collected females, as well as ovipositional response to physical characteristics of the host plant. Both mated and unmated females produced eggs, but at different rates. Field-collected females and mated females reared in the laboratory laid similar numbers of egg masses containing similar numbers of eggs, but unmated females laid approximately one half as many eggs, the result primarily of smaller clutch size. Mated females reared in the laboratory had a preovipositional period of 11.4 +/- 1.7 versus 29 + 11.7 d for unmated females, and unmated females lived significantly longer than mated females. The angle and diameter of stems of V. trilobum, a very susceptible host, both greatly influenced oviposition; females laid most eggs on vertically oriented stems, and those of smallest diameter; when these factors were combined, stem diameter predominated. Females also had a very strong geotactic response, preferring to lay eggs on portions of stems toward gravity, even when stems were at fairly shallow angles. PMID- 18419926 TI - Larval feeding on Bt hybrid and non-Bt corn seedlings by Harmonia axyridis (Coleoptera: Coccinellidae) and Coleomegilla maculata (Coleoptera: Coccinellidae). AB - Zoophytophagy is an omnivorous activity that occurs when a primarily carnivorous species feeds on plant material. Plant feeding by beneficial predators may have negative consequences if the plant material has been chemically treated, contains toxins, or was transgenically altered. Although common in predaceous Hemiptera, zoophytophagy has been rarely studied in aphidophagous coccinellids. This study examined the likelihood of feeding on Bt and non-Bt corn seedlings by third- and fourth-instar coccinellid larvae, the regularity of feeding events by fourth instars, and the effect of leaf feeding on development time and adult size. Both third- and fourth-instar Harmonia axyridis Pallas and Coleomegilla maculata DeGeer consume leaf tissue, with fourth instars being significantly more likely to feed on corn seedlings. C. maculata larvae ingested leaf tissue more frequently than H. axyridis. Furthermore, when given access to corn seedlings daily, development time of fourth-instar C. maculata increased after Bt hybrid corn treatments compared with non-Bt corn treatments. Zoophytophagous feeding behavior is thought to sustain predators during times of low prey availability, and leaf tissue feeding by coccinellids has typically been attributed to their need for water. However, in this study, tissue feeding regularly occurred even though coccinellid larvae had constant access to water and a daily ad libitum supply of aphids. We suggest that, in addition to environmental conditions, the physiological state of the zoophytophagous species will influence the probability of plant feeding. PMID- 18419927 TI - Repellency of a wireworm, Agriotes obscurus (Coleoptera: Elateridae), on exposure to synthetic insecticides in a soil-less bioassay. AB - A soil-less bioassay arena to test repellency of wireworms (A. obscurus) to insecticides and carrier solvents is described. The bioassay and variables measured distinguish between shorter-range (contact and/or volatile) and longer range (volatile) repellency. Wireworm positions are recorded every 3 s for 20 min, and average speed, rate of slowing, and longer- and shorter-range repellent behaviors calculated. Shorter-range repellency is determined with a Wireworm Repellency Score (WRS, range 0-100), calculated before contact and after contact with test chemicals. Of two carrier solvents tested, wireworms were strongly repelled by acetone (WRS = 57) but not by water (WRS = 1) when introduced to the bioassay arenas immediately after chemical inoculation. When bioassay arenas were assembled/sealed 2 min after inoculation, acetone elicited no repellency (WRS = 2). When dissolved in acetone in bioassays assembled with a 2-min delay, imidacloprid, chlorpyrifos, lindane, and tefluthrin elicited slight to moderate repellency at the highest concentrations tested (WRS = 30, 48, 42, and 49, respectively). Both longer- and shorter-range repellency increased over the duration of the observation period for lindane and tefluthrin, and shorter-range repellency also increased over the duration of the observation period for chlorpyrifos. Removal of volatiles in the bioassay arena by vacuum considerably affected wireworm movement in the arena, with the repellency elicited by acetone and lindane being significantly reduced. Clothianidin elicited no longer- or shorter-range repellency. PMID- 18419928 TI - Using reports of bee mortality in the field to calibrate laboratory-derived pesticide risk indices. AB - Mounting evidence suggests that pollinators worldwide are experiencing dramatic population declines, and exposure to pesticides is one of the factors that can account for this. By making use of a database containing more than two decades of honey bee (Apis mellifera) hive poisoning incidents from the United Kingdom (Wildlife Incident Investigation Scheme [WIIS]) and corresponding pesticide use surveys, we attempted to explain honey bee poisoning incidents in the field using models derived from pesticide use information, laboratory-generated bee toxicity data (defined as a hazard ratio; application rate divided by LD(50)), and physico chemical properties of the applied pesticides. Logistic regression analyses were used to assess the relationship between honey bee poisoning incidents in the field and these parameters. In analyzing models with multiple dimensions, we selected the best model by the best subset method, an iterative method based on maximum likelihood estimation, and Akaike's information criterion. Results suggested that the size of the area treated and hazard ratios calculated from application rates and oral or contact toxicity (but the latter especially) can be used to predict the likelihood that honey bee mortality will occur. Model predictions also suggest that some insecticides carry an extreme risk for bees, despite the lack of documented incidents. PMID- 18419929 TI - Seasonal occurrence of key arthropod pests and associated natural enemies in Alabama Satsuma citrus. AB - Six Alabama Satsuma mandarin orchards (four conventionally sprayed and two unsprayed) were surveyed during 2005 and 2006 to determine the population dynamics of arthropod pests and their natural enemies. Twenty-eight arthropod pest species were encountered; the major foliage pests were citrus whitefly, Dialeurodes citri (Ashmead); purple scale, Lepidosaphes beckii (Newman); Glover scale, L. gloveri (Packard); and citrus red mite, Panonychus citri (McGregor). Two distinct population peaks were recorded for citrus whitefly at most locations. The most important direct sources of citrus whitefly mortality were parasitism by Encarsia lahorensis (Howard) and infection by the pathogenic fungus, Aschersonia aleyrodis Webber. In general, all stages of both scale insects (purple scale and Glover scale) were present in the orchards year-round, indicative of overlapping generations; however, the highest densities were recorded during the early season. Citrus whitefly, purple scale, and Glover scale were more abundant on leaves collected from the interior of the tree canopy than in the exterior canopy. Citrus red mite densities were highest in the spring, with populations declining at the start of the summer, and were more abundant in the exterior canopy than in the interior canopy. The most important natural enemies of citrus red mite were predatory mites belonging to several families, of which Typhlodromalus peregrinus Muma (Phytoseiidae) was the predominant species. Major differences were recorded in the relative abundance of different arthropod pest species in the orchards: citrus whitefly, purple scale, and Glover scale predominated in the unsprayed orchards, whereas citrus red mite infestations were more severe in the sprayed orchards. The results are discussed in relation to the possible effect of orchard management practices on abundance of the major pests. PMID- 18419930 TI - Influence of host plant stages on carob moth (Lepidoptera: Pyralidae) development and fitness. AB - Different generations of the carob moth, Ectomyelois ceratoniae (Zeller), use different date, Phoenix dactylifera L., fruit stages as they become available during the summer months in southern California. These are the kimri, khalal, and tamar fruit stages. This study was conducted to determine whether carob moth development and fitness were affected by these different fruit stages. Developmental time from neonate larvae to adult, when reared at 31.9 degrees C and 82.1% RH, ranged from 30.5 to 32.3 d for females and 27.1 to 29.5 d for males on the different field-collected fruit stages. Males and females had the highest emergent weight when reared as larvae on kimri fruit and the lowest on tamar fruit. Females laid the most eggs when reared on kimri fruit and the least when reared on tamar fruit. Estimates of population doubling times ranged from 5.4 d on artificial diet (included as a control) to 7.5 d on tamar fruit. This short doubling time shows the ability of carob moth to develop rapidly under optimal conditions. Degree-day (DD) estimates for carob moth development ranged from 636 DD on kimri fruit to 658 DD on tamar fruit, which translate to 32-50 d under field temperatures in the area where dates are grown. Potential implications for field management of E. ceratoniae include improved timing of insecticide treatments to limit population growth early in the season rather than the conventional late season approach. PMID- 18419931 TI - Influence of fertilization on the Colorado potato beetle, Leptinotarsa decemlineata, in organic potato production. AB - The abundance of the Colorado potato beetle, Leptinotarsa decemlineata (Say), in organically grown potato did not change significantly in response to increasing rates of dehydrated poultry manure. However, peaks of abundance of larvae were shifted forward in time in response to the high rate of organic fertilizer. Tests using excised foliage showed that the shift was not caused by differential larval mortality or longer developmental times. Time allocation to resting, walking, and feeding by adults was similar regardless of fertilizer rate. Adult foliage consumption was unaffected by organic fertilizer rates in no choice tests and significantly affected in few choice tests. A 22% longer larval development time on plants treated with low fertilizer rate than on plants with high rate was the most significant effect. Even though maximum plant height, canopy, biomass, and yield were significantly smaller in the organic than in conventional plots, the suitability of the plants was not affected except for reduced feeding by summer beetles. Summer adults spent less time feeding and consumed two to five times less foliage on organic potato than on inorganically fertilized and conventionally produced plants. The overall influence of fertilizer on Colorado potato beetle populations was limited and therefore can only play a secondary role in management strategies for organic potato. Avoidance of excessive organic fertilizer that promotes short larval development time and extension of the period over which large Colorado potato beetle larvae are present should be recommended. PMID- 18419932 TI - Biology and biotype determination of greenbug, Schizaphis graminum (Hemiptera: Aphididae), on seashore paspalum turfgrass (Paspalum vaginatum). AB - Greenbug, Schizaphis graminum (Rondani) (Hemiptera: Aphididae), was first discovered damaging seashore paspalum (Paspalum vaginatum Swartz) turfgrass in November 2003 at Belle Glade, FL. Inquiries to several golf courses with seashore paspalum turf across southern Florida indicated infestation was wide spread by April 2004. Damage symptoms progress from water soaked lesions surrounding feeding sites within 24 h to chlorosis and necrosis of leaf tips within 96 h. Problems caused by greenbug feeding were initially misdiagnosed as fertilizer, disease, other insects, or water management problems because aphids were not previously found on warm season turfgrasses. Greenbug development and fecundity studies were conducted on six seashore paspalum varieties: 'Aloha,' 'SeaDwarf,' 'SeaGreen,' 'SeaIsle,' 'SeaWay,' and 'SeaWolf.' Greenbug did not survive on 'SeaWolf.' Development rates (mean +/- SEM) ranged from 7.6 +/- 0.2 to 8.2 +/- 0.2 d on the remaining varieties. Greenbug longevity and fecundity on 'Aloha' were significantly less than on the other varieties. The estimated intrinsic rate of natural increase (r(m)) for greenbug ranged from 0.24 to 0.26 across tested varieties. Values for net reproductive rate (R(o)) ranged from 12.3 on 'Aloha' to 40.4 on 'SeaWay.' In feeding trials on indicator plants, the Florida isolate of greenbug exhibited a unique biotypic profile most commonly found on noncultivated grass hosts. It was virulent on the wheat variety GRS1201 that is resistant to the principal agricultural biotypes attacking small grains and to all currently available resistant sorghum varieties. PMID- 18419933 TI - Effect of an alternate weed host, hairy nightshade, Solanum sarrachoides, on the biology of the two most important potato leafroll virus (Luteoviridae: Polerovirus) vectors, Myzus persicae and Macrosiphum euphorbiae (Aphididae: Homoptera). AB - Hairy nightshade, Solanum sarrachoides (Sendtner), is a ubiquitous weed in potato agro-ecosystems and nonagricultural lands of southeastern Idaho and the Pacific Northwest. This weed increases the complexity of the Potato leafroll virus (PLRV) (Luteoviridae: Polervirus)-potato pathosystem by serving as aphid and virus reservoir. Previous field studies showed higher densities of green peach aphid, Myzus persicae (Sulzer), and potato aphid, Macrosiphum euphorbiae (Thomas), the two most important vectors of PLRV, on S. sarrachoides compared with potato plants in the same fields. Some of the S. sarrachoides plants sampled in these surveys tested positive for PLRV. Viral infections can alter the physiology of plant hosts and aphid performance on such plants. To understand better the potential effects of S. sarrachoides on the PLRV-potato-aphid pathosystem, the life histories of M. persicae and M. euphorbiae were compared on virus-free and PLRV-infected S. sarrachoides and potato. Individual nymphs of each aphid species were held in clip cages on plants from each treatment to monitor their development, survival, and reproductive output. Nymphal survival for both aphids across plant species was higher on S. sarrachoides than on potato, and, within plant species, it was higher on PLRV-infected plants than on noninfected plants. With a few exceptions, similar patterns occurred for fecundity, reproductive periods, adult longevity, and intrinsic rate of increase. The enhanced performance of aphids on S. sarrachoides and on PLRV-infected plants could alter the vector population dynamics and thus the PLRV-disease epidemiology in fields infested with this weed. PMID- 18419934 TI - Longevity and fecundity of Japanese beetle (Popillia japonica) on foliage grown under elevated carbon dioxide. AB - Atmospheric levels of carbon dioxide (CO(2)) have been increasing steadily over the last century. Plants grown under elevated CO(2) experience physiological changes that influence their suitability as food. Previous studies have found increased insect herbivory on plants grown under elevated CO(2). To determine effects of consuming foliage of soybean (Glycine max) grown under elevated CO(2) on adult survivorship and fecundity, Japanese beetles (Popillia japonica Newman) were fed for the duration of their adult lives leaves grown under elevated CO(2) (550 mumol/mol), under ambient atmosphere (370 mumol/mol), or grown under ambient atmosphere but supplemented with a solution of sugars. To determine effects of a diet of foliage grown under elevated ozone (O(3)), another anthropogenic gaseous pollutant, beetles in the laboratory were fed soybean leaves grown under elevated CO(2), elevated O(3), or a combination of both elevated gases. Leaf tissue was also analyzed for longevity-enhancing antioxidants, because increases in dietary antioxidants can increase lifespan. Lifespan of Japanese beetles was prolonged by 8-25% when fed foliage developed under elevated CO(2), but consuming foliage that had taken up sugars to approximately the same level as foliage grown under elevated CO(2) had no effect on fecundity or longevity. Females consuming elevated CO(2) foliage laid approximately twice as many eggs as females fed foliage grown under ambient conditions. Consuming foliage grown under elevated O(3) had no effect on fecundity. No significant differences in total antioxidant content of foliage from ambient and elevated CO(2) conditions were detected. Although the precise mechanism is unclear, by altering components of leaf chemistry other than sugar content, elevated CO(2) may increase populations of Japanese beetles and their impact on crop productivity. PMID- 18419935 TI - Japanese beetles facilitate feeding by green june beetles (Coleoptera: Scarabaeidae) on ripening grapes. AB - Adult activity of native green June beetles, Cotinis nitida L., and invasive Japanese beetles, Popillia japonica Newman, peaked in late July in Kentucky coincident with later stages of veraison in early- and midseason ripening grape (Vitis spp.) cultivars. Most C. nitida feeding aggregations sampled on clusters of early-ripening grapes in the research vineyard also contained Japanese beetles. Assays showed C. nitida generally are unable to bite into intact ripe grape berries, whereas Japanese beetles readily do so. The beetles' disparate biting ability is likely because of differences in their mandibles, which are sharply pointed and opposable in P. japonica compared with C. nitida mandibles that are bluntly spatulate, do not meet at their tips, and seemingly are only suited for feeding on fruit pulp or other soft food. Japanese beetles were shown to facilitate C. nitida feeding by biting through the skin and providing access to the soft berry pulp. Juice from early- and midseason ripening grape cultivars with relatively high sugar content elicited the greatest feeding by P. japonica. A scenario is suggested wherein Japanese beetles open wound sites and contaminate fruits with yeasts that induce fermentation volatiles that attract C. nitida. Japanese beetles had difficulty biting berries of Sunbelt, a late-ripening cultivar. Phenological resistance, i.e., planting cultivars that ripen after peak flight, could be an effective management strategy. Establishment of P. japonica in grape-growing regions of the southeastern United States will likely elevate the pest status of C. nitida in vineyards. PMID- 18419936 TI - Evaluating popcorn as a potential refuge of Ostrinia nubilalis (Lepidoptera: Crambidae). AB - Popcorn was evaluated in a series of experiments conducted over four growing seasons for its potential as a refuge for European corn borer, Ostrinia nubilalis (Hubner). Objectives of these studies were to determine whether more larvae were produced in popcorn than in field corn and to determine how popcorn influenced female oviposition and larval distribution in neighboring field corn. Two varieties of popcorn (M140, 105d and M3374Y, 118d), one mixture of popcorn (50% 105d and 50% 118d), and field corn (DK580, 108d) were evaluated. Number of egg masses, eggs per egg mass, and larvae were significantly higher in popcorn compared with field corn. Moth oviposition and larval distribution were evaluated using 105d popcorn embedded in several cornfields across Iowa. The row of field corn adjacent to popcorn had significantly more larvae compared with background field corn. In larger field experiments, O. nubilalis larval survival after overwintering was significantly different, with 2.2-18.7 times more O. nubilalis larvae surviving in popcorn than field corn. The potential use of popcorn as an O. nubilalis refuge for genetically engineered corn is considered. PMID- 18419937 TI - Nano-fiber scaffold science and tissue engineering. Foreword. PMID- 18419938 TI - Integrating novel technologies to fabricate smart scaffolds. AB - Tissue engineering aims at restoring or regenerating a damaged tissue by combining cells, derived from a patient biopsy, with a 3D porous matrix functioning as a scaffold. After isolation and eventual in vitro expansion, cells are seeded on the 3D scaffolds and implanted directly or at a later stage in the patient's body. 3D scaffolds need to satisfy a number of requirements: (i) biocompatibility, (ii) biodegradability and/or bioresorbability, (iii) suitable mechanical properties, (iv) adequate physicochemical properties to direct cell material interactions matching the tissue to be replaced and (v) ease in regaining the original shape of the damaged tissue and the integration with the surrounding environment. Still, it appears to be a challenge to satisfy all the aforementioned requisites with the biomaterials and the scaffold fabrication technologies nowadays available. 3D scaffolds can be fabricated with various techniques, among which rapid prototyping and electrospinning seem to be the most promising. Rapid prototyping technologies allow manufacturing scaffolds with a controlled, completely accessible pore network--determinant for nutrient supply and diffusion--in a CAD/CAM fashion. Electrospinning (ESP) allows mimicking the extracellular matrix (ECM) environment of the cells and can provide fibrous scaffolds with instructive surface properties to direct cell faith into the proper lineage. Yet, these fabrication methods have some disadvantages if considered alone. This review aims at summarizing conventional and novel scaffold fabrication techniques and the biomaterials used for tissue engineering and drug delivery applications. A new trend seems to emerge in the field of scaffold design where different scaffolds fabrication technologies and different biomaterials are combined to provide cells with mechanical, physicochemical and biological cues at the macro-, micro- and nano-scale. If merged together, these integrated technologies may lead to the generation of a new set of 3D scaffolds that satisfies all of the scaffolds' requirements for tissue-engineering applications and may contribute to their success in a long-term scenario. PMID- 18419939 TI - Electrohydrodynamic atomization: a versatile process for preparing materials for biomedical applications. AB - Electrohydrodynamic atomization phenomena have increasingly attracted the attention of researchers who are interested in building micro- or nanometer architectures, such as fibers and encapsulated particles with a controllable microstructure. There are two main electrohydrodynamic atomization techniques: electrospraying and electrospinning. These techniques are unique processes in that they produce fibers and particles with dimensions that range from micrometers to nanometers depending on the processing parameters. This paper presents the principles, processes and potential biomedical applications for electrospraying and electrospinning of biomaterials. Both of these techniques offer great research opportunities. Emphasis will focus on the effects of processing parameters of electrohydrodynamic atomization on morphologies and microstructure of the final products. In addition, some potential applications are proposed based on the remarkable characteristics of the biomaterials generated using electrospraying and electrospinning. PMID- 18419940 TI - Measuring fiber alignment in electrospun scaffolds: a user's guide to the 2D fast Fourier transform approach. AB - In this study we describe how to use a two-dimensional fast Fourier transform (2D FFT) approach to measure fiber alignment in electrospun materials. This image processing function can be coupled with a variety of imaging modalities to assign an objective numerical value to scaffold anisotropy. A data image of an electrospun scaffold is composed of pixels that depict the spatial organization of the constituent fibers. The 2D FFT function converts this spatial information into a mathematically defined frequency domain that maps the rate at which pixel intensities change across the original data image. This output image also contains quantitative information concerning the orientation of objects in a data image. We discuss the theory and practice of using the frequency plot of the 2D FFT function to measure relative scaffold anisotropy and identify the principal axis of fiber orientation. We note that specific degrees of scaffold anisotropy may represent a critical design feature in the fabrication of tissues that will be subjected to well-defined uniaxial mechanical loads. This structural property may also represent a source of guidance cues that can be exploited to regulate cell phenotype. PMID- 18419941 TI - Characterization of neural stem cells on electrospun poly(epsilon-caprolactone) submicron scaffolds: evaluating their potential in neural tissue engineering. AB - Development of biomaterials with specific characteristics to influence cell behaviour has played an important role in exploiting strategies to promote nerve regeneration. The effect of three-dimensional (3D) non-woven electrospun poly(epsilon-caprolactone) (PCL) scaffolds on the behaviour of rat brain-derived neural stem cells (NSCs) is reported. The interaction of NSCs on the randomly orientated submicron (PCL) fibrous scaffolds, with an average fibre diameter of 750 +/- 100 nm, was investigated. The PCL scaffolds were modified with ethylenediamine (ED) to determine if amino functionalisation and changes in surface tension of the fibrous scaffolds affected the proliferation and differentiation characteristics of NSCs. Surface tension of the fibrous scaffold increased upon treatment with ED which was attributed to amine moieties present on the surface of the fibres. Although surface treatment did not change the differentiation of the NSCs, the modified scaffolds were more hydrophilic, resulting in a significant increase in the number of adhered cells, and increased spreading throughout the entirety of the scaffold. When the NSCs were seeded on the PCL scaffolds in the presence of 10% FBS, the stem cells differentiated primarily into oligodendrocytes, indicating that electrospun PCL has the capacity to direct the differentiation of NSCs towards a specific lineage. The data presented here is useful for the development of electrospun biomaterial scaffolds for neural tissue engineering, to regulate the proliferation and differentiation of NSCs. PMID- 18419942 TI - Hybrid nanofibrous scaffolds from electrospinning of a synthetic biodegradable elastomer and urinary bladder matrix. AB - Synthetic materials can be electrospun into submicron or nanofibrous scaffolds to mimic extracellular matrix (ECM) scale and architecture with reproducible composition and adaptable mechanical properties. However, these materials lack the bioactivity present in natural ECM. ECM-derived scaffolds contain bioactive molecules that exert in vivo mimicking effects as applied for soft tissue engineering, yet do not possess the same flexibility in mechanical property control as some synthetics. The objective of the present study was to combine the controllable properties of a synthetic, biodegradable elastomer with the inherent bioactivity of an ECM derived scaffold. A hybrid electrospun scaffold composed of a biodegradable poly(ester-urethane)urea (PEUU) and a porcine ECM scaffold (urinary bladder matrix, UBM) was fabricated and characterized for its bioactive and physical properties both in vitro and in vivo. Increasing amounts of PEUU led to linear increases in both tensile strength and breaking strain while UBM incorporation led to increased in vitro smooth muscle cell adhesion and proliferation and in vitro mass loss. Subcutaneous implantation of the hybrid scaffolds resulted in increased scaffold degradation and a large cellular infiltrate when compared with electrospun PEUU alone. Electrospun UBM/PEUU combined the attractive bioactivity and mechanical features of its individual components to result in scaffolds with considerable potential for soft tissue engineering applications. PMID- 18419943 TI - Silk-based electrospun tubular scaffolds for tissue-engineered vascular grafts. AB - Electrospinning was used to fabricate non-woven nanofibrous tubular scaffolds from Bombyx mori silk fibroin using an all aqueous process. Cell studies and mechanical characterization tests were performed on the electrospun silk tubes to assess the viability of their usage in bioengineering small-diameter vascular grafts. Human endothelial cells and smooth muscle cells were successfully cultured on the electrospun silk. Mechanical characterization tests demonstrated burst strength sufficient to withstand arterial pressures and tensile properties comparable to native vessels. These cellular and mechanics outcomes demonstrate potential utility of these electrospun silk scaffolds for small-diameter vascular grafts. PMID- 18419944 TI - Tuning the mechanical and bioresponsive properties of peptide-amphiphile nanofiber networks. AB - Here we describe peptide amphiphiles (PAs) which can be self-assembled into nanofiber networks using divalent ions. These networks possess several key properties of extracellular matrix (ECM) including cell-adhesive ligands, enzyme mediated degradation and self-assembly into hierarchical organization. The self assembly of PAs and growth of nanofibers could be controlled by modifications of the chemical structure of the PA and/or addition of divalent ions. Altering the length of PAs alters the viscoelastic properties and degradation kinetics of nanofiber networks. Neural cells were successfully encapsulated within nanofiber networks by self-assembly of PAs. Cell adhesive ligands containing nanofiber networks supported neural cells growth, and their cellular behaviors depended on the concentration of cell adhesive ligands. Therefore, we have demonstrated that mechanical properties, degradability, and bioactivity of nanofiber networks could be tuned by altering the chemical composition and the length of PAs. PMID- 18419945 TI - Electrospun chitosan-P(LLA-CL) nanofibers for biomimetic extracellular matrix. AB - Chitosan-poly(L-lactic acid-co-epsilon-caprolactone)(50:50) (P(LLA-CL)) (CS/P(LLA CL)) blends were electrospun into nanofibers using 1,1,1,3,3,3-hexafluoro-2 propanol (HFIP) and trifluoroacetic acid (TFA) as solvents. Chitosan, which is difficult to electrospin into nanofibers, could be easily electrospun into nanofibers with addition of a small portion of P(LLA-CL). The fiber diameter depended on both the polymer concentration and the blend ratio of chitosan to P(LLA-CL). The average fiber diameter increased with increasing polymer concentration and decreasing the blend ratio of chitosan to P(LLA-CL). X-ray diffractometry (XRD) and Fourier-transform infrared (FT-IR) spectra were measured to characterize blended nanofibers. The porosity of CS/P(LLA-CL) nanofiber mats increased with increasing the weight ratio of chitosan to P(LLA-CL), while both the tensile strength and the ultimate strain increased with increasing P(LLA-CL) ratio. Fibroblast cell growth on nanofiber mats were investigated with MTT assay and scanning electron microscope (SEM) observation. The highest cell proliferation was observed on the nanofiber mats when the weight ratio of chitosan to P(LLA-CL) was 1:2. As SEM images shown, fibroblast cells showed a polygonal shape on blend nanofiber mats and migrated into the nanofiber mats. PMID- 18419947 TI - [Inflection of green channels in emergency cardiovascular care]. PMID- 18419946 TI - In vitro and in vivo analysis of co-electrospun scaffolds made of medical grade poly(epsilon-caprolactone) and porcine collagen. AB - In this study, a nanofiber mesh made by co-electrospinning medical grade poly(epsilon-caprolactone) and collagen (mPCL/Col) was fabricated and studied. Its mechanical properties and characteristics were analyzed and compared to mPCL meshes. mPCL/Col meshes showed a reduction in strength but an increase in ductility when compared to PCL meshes. In vitro assays revealed that mPCL/Col supported the attachment and proliferation of smooth muscle cells on both sides of the mesh. In vivo studies in the corpus cavernosa of rabbits revealed that the mPCL/Col scaffold used in conjunction with autologous smooth muscle cells resulted in better integration with host tissue when compared to cell free scaffolds. On a cellular level preseeded scaffolds showed a minimized foreign body reaction. PMID- 18419948 TI - [Midazolam attenuates mental stress in conscious patients during cardiopulmonary resuscitation performed next bed in intensive care unit]. AB - OBJECTIVE: To evaluate the mental stress level and alterations in circulatory physiology in conscious patients during cardiopulmonary resuscitation (CPR) performed next bed in intensive care unit (ICU), and to investigate the possible effective interventions. METHODS: Eighty-seven conscious patients, selected consecutively from June 2003 to September 2006, were randomly allocated into control group (received normal saline), psychological nursing group (received psychological nursing intervention) or sedation group (received midazolam 0.1 mg/kg intravenous injection based on psychological nursing intervention) when CPR was performed in our ICU. Plasma concentrations of norepinephrine, epinephrine, cortisone and glucose were analyzed at the time points of beginning of CPR, 10 minutes, 4 and 24 hours after CPR in the first 40 patients. Heart rate (HR), systolic blood pressure (SBP), mean arterial pressure (MAP) and arrhythmia within 24 hours after CPR were recorded in all patients. RESULTS: Plasma levels of norepinephrine, epinephrine and cortisone were significantly increased at 10 minutes after CPR and persisted for 4 hours in 13 patients of the control group (P<0.05 or P<0.01). Though with the similar tendency, significant increase of cortisone level was observed in 13 patients who had received psychological nursing intervention (P<0.05 or P<0.01). The analyzed stress hormones showed little variation in 14 patients who were given midazolam at 10 minutes and 4 hours after CPR. Notably, 24 hours after CPR, they were decreased below the levels which were observed at the beginning of CPR (all P<0.01). Blood glucose levels were markedly higher in both control and psychological nursing groups than the level in sedation group within 24 hours. HR was accelerated 10 minutes after CPR, SBP was significantly increased, the incidence rate of arrhythmia was high (84.6%, 22/26; 54.5%, 18/33) in the non-sedation groups. Circulatory physiological alterations were least marked in sedation group (21.4%, 6/28, both P<0.01). CONCLUSION: Mental stress is significantly heightened in conscious patients during CPR performed next bed in ICU, and it induces severe circulatory physiological alterations. Psychological nursing alone is not affective in alleviating this acute mental stress. However, low dose of midazolam is found to be an effective intervention. PMID- 18419949 TI - [Effect of urapidil on heart function and ventricular systolic synchrony in acute myocardial infarction patients with no-reflow phenomenon after percutaneous coronary intervention]. AB - OBJECTIVE: To evaluate the influence of intracoronary administration of urapidil on myocardial blush grade (MBG) and left ventricular systolic function and synchrony in the acute myocardial infarction (AMI) patients with no-reflow phenomenon after percutaneous coronary intervention (PCI) identified by MBG. METHODS: Forty-three patients with AMI, in whom primary PCI was successfully performed (6.25+/-2.37) hours after the onset of angina pectoris,were found to have no-reflow phenomenon. They were randomized into two groups: urapidil group (n=22) and no-reflow control group (n=21). Nitroglycerin (200 microg) was injected into coronary artery. Urapidil (5 mg) was injected into coronary artery after 10 minutes in the urapidil group, and 0.9% NaCl (5 ml, weight percentage) was injected into coronary artery in the no-reflow control group. All the patients received same standard therapy afterwards. The left ventriculography (LVG) was performed immediately and 6 months after PCI to measure the ventricular volume, left ventricular end-diastolic pressure (LVEDP), and wall motion score (WMS). Equilibrium radionuclide angiography (ERNA) was performed 1 week and 6 months after PCI to determine the parameters of left ventricular systolic function and systolic synchrony. RESULTS: The MBG of urapidil group and control group was grade 0.77+/-0.31 and grade 0.77+/-0.28 after PCI, respectively. The MBG remained unchanged in control group and significantly increased from grade 0.77+/-0.31 to grade 2.37+/-0.27 10 minutes in urapidil group (P<0.05). Follow-up at 6 months after AMI-PCI, left ventricular end-systolic volume index (LVESVI), left ventricular end-diastolic volume index (LVEDVI), WMS and LVEDP were significantly lower in urapidil group compared with those in control group respectively (all P<0.05). The values of left ventricular ejection fraction (LVEF), peak ejection rate (PER), peak filling rate (PFR) of the ERNA as measured by ERNA were significantly increased in urapidil group compared with that in control group (all P<0.05). Phase analysis showed that the left ventricular systolic synchrony parameters phase shift (PS), full width at half maximum (FWHM) and peak phase standard deviation (PSD) were also significantly lower in urapidil group than those in control group (all P<0.05). CONCLUSION: Intracoronary administration of urapidil can attenuate the no-reflow phenomenon, improve the left ventricular systolic function and synchrony in patients with no-reflow phenomenon after AMI-PCI. PMID- 18419950 TI - [Impact of atrial fibrillation on prognosis of chronic heart failure patients with left ventricular ejection fraction>or=0.5]. AB - OBJECTIVE: To explore the prevalence, distribution type and impact of atrial fibrillation on prognosis of hospitalized patients with congestive heart failure (CHF) with left ventricular ejection fraction (LVEF)>or=0.50. METHODS: The medical records of 417 unselected consecutive patients with CHF were retrospectively reviewed. Patients were categorized as LVEF<0.50 or LVEF>or=0.50. And they were also categorized by the past history of atrial fibrillation and divided into three groups: paroxysmal atrial fibrillation, continuous atrial fibrillation and onset of atrial fibrillation after admission. Then the vicious events, the number of readmission due to CHF and the interval between discharge and readmission were observed and recorded. RESULTS: Male patients were prevalent with CHF whose LVEF<0.50. The occurrence of acute myocardial infarction in the 1st year [15.6% (34/218)] was higher than that of CHF with LVEF>or=0.5 [8.0% (16/199), P<0.01]. The occurrence of cerebral stroke in patients with atrial fibrillation [24.3% (27/111)] was higher than that of patients without atrial fibrillation [8.4% (9/107), P<0.05). The numbers of acute coronary syndrome and cardiac death were also increased. In the patients with CHF whose LVEF>or=0.50 the incidence atrial fibrillation occurring after readmission was significantly higher than that of patient with CHF whose LVEF<0.50 (51 vs. 30, P<0.05). Atrial fibrillation could be found in nearly 2/3 of patients. And the number of readmission (2.78+/-1.79 vs. 2.00+/-1.35, P<0.01) was increased, while the interval between discharge and readmission [(117+/-107) days vs. (154+/-130) days, P<0.05] was shorter. CONCLUSION: Atrial fibrillation occurs more likely in patients with CHF whose LVEF>or=0.50, leading to a shorter interval of readmission. Therefore the importance of treatment of atrial fibrillation should be emphasized. PMID- 18419951 TI - [Brain natriuretic peptide and left ventricular remodeling after emergency percutaneous coronary intervention in acute myocardial infarction patients]. AB - OBJECTIVE: To investigate the effect of emergency percutaneous coronary interventional (PCI) treatment on plasma brain natriuretic peptide (BNP) levels and left ventricular remodeling in patients with acute myocardial infarction (AMI). METHODS: This study included 118 patients with AMI and 20 healthy volunteers (their results were regarded as normal reference). Fifty-two patients who underwent successful emergency PCI 6-12 hours after onset were named as PCI group, and 66 patients rejected or in whom emergency PCI failed served as the control group. Plasma BNP levels were determined with Triage rapid assay at admission,at 12, 24, 48, 72 hours and 7, 14, 28 days after admission for both groups. Left ventricular ejection function (LVEF) was assessed by echocardiography with the modified Simpson's equation on 3-5 days and 28 days. Same assay was performed for 20 healthy volunteers. RESULTS: Plasma BNP levels of both groups were significantly higher at admission than those of volunteers. There was significant difference in BNP levels between two groups at corresponding time points (all P<0.01). In PCI group, BNP level peaked during 12 24 hours after admission, whereas two peaks of elevation of BNP levels were detected in control group, the first peak appeared during 12-24 hours and the second peak on 7 days after admission. Plasma BNP levels in PCI group were significantly lower than those of control group at corresponding time points (all P<0.01). There was no difference in LVEF level between two groups on 3-5 days after admission. LVEF level after emergency PCI was significantly higher than that of control group on 28 days after admission (P<0.01). CONCLUSION: Emergency PCI lowers plasma BNP level and improve LVEF level in patients with AMI, and it may reverse ventricular remodeling. PMID- 18419952 TI - [Method of anesthesia and perioperative management for off-pump coronary artery bypass grafting]. AB - OBJECTIVE: To investigate the method of anesthesia and perioperative management for off- pump coronary artery bypass grafting (OPCABG), and to examine the hemodynamic changes during the operation. METHODS: One hundred and thirteen patients undergoing OPCABG were studied. Anesthesia induction was achieved by giving midazolam 0.05 mg/kg, ketamine 0.5 mg/kg, vecuronium 0.13-0.15 mg/kg and fentanyl 5-10 microg/kg. Vecuronium and fentanyl were given as needed, isoflurane and propofol were administered for maintenance. Hemodynamic indexes, ST-segment change in electrocardiogram (ECG) and lidocaine's influence to the cardiac rhythm were observed and recorded during the main process of operation. RESULTS: Observation before and during the operation showed that mean arterial pressure (MAP) and heart rate (HR) decreased a little but within clinical acceptable range, at the beginning, and it recovered soon. The result of rate-pressure product (RPP) showed that the reduction of cardiac muscle oxygen consumption was satisfactory. There was no significant change in ST-segment in ECG (P>0.05) before and during the main procedure. Giving 1 mg/kg lidocaine before the operation could obviously lower the possibility or degree of premature ventricular extrasystole. CONCLUSION: OPCABG can be performed safely under the above method of anesthesia. The perioperative management is effective and satisfactory in maintaining stable circulation and in keeping oxygen supply and consumption balance. PMID- 18419953 TI - [Expression of renal aquaporin 2 after circulatory arrest]. AB - OBJECTIVE: To investigate the inhibition of renal aquaporin 2 (AQP2) and AQP2 mRNA expression due to circulatory arrest. METHODS: Forty rats were randomized into two groups. The circulatory arrest-resuscitation model was reproduced in resuscitation group (20 rats). Rats in control group were subjected to the same procedure but only received mechanical ventilation without asphyxia and resuscitation (20 rats). Rats in each group were divided into 1-hour group and 3 hour group randomly (10 rats in each group). Serum creatinine (SCr) and blood urine nitrogen (BUN) concentrations were determined in every rat. Medulla of kidney in every rat was isolated for the determination of AQP2 and AQP2 mRNA expression with Western blotting and reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: There were no differences in resuscitation time, SCr and BUN concentrations between resuscitation and control groups. The AQP2 protein and mRNA expression in resuscitation 1-hour and 3-hour groups were significantly decreased compared with those in control group at the same time points [protein: 38.35+/-2.08 vs. 41.06+/-1.04 (1-hour), 31.89+/-1.57 vs. 41.45+/-0.58 (3-hour); mRNA: 0.61+/-0.13 vs. 0.87+/-0.14 (1-hour),0.54+/-0.11 vs. 0.85+/-0.12 (3-hour), all P<0.01], and they were further lowered in resuscitation 3-hour group (both P<0.01). CONCLUSION: Circulatory arrest would lead to down regulation of renal AQP2 and AQP2 mRNA expression. These changes appear earlier than those of SCr and BUN. PMID- 18419955 TI - [Effects of different delivery routes of CD34+ stem cells on cardiac function in the ischemic cardiomyopathy of rats]. AB - OBJECTIVE: To compare the effects of CD34-positive stem cells delivered by different routes on cardiac function in rats with ischemic cardiomyopathy, and to evaluate the efficacy and safety of stem cell transplantation. METHODS: Sixty four male Wistar rats were randomized into cell infusion group (n=30), acute myocardial infarction (AMI) model group (n=20) and sham operation group (n=14). AMI model was reproduced by ligation of left anterior descending coronary artery. CD34+ stem cells mobilized with granulocyte-colony stimulating factor (G-CSF) were purified by immunomagnetic beads in 30 donor rats. Seven days after the injury about (7-9)x10(7) CD34+ stem cells were infused through the femoral vein or through epicardium of recipient rats respectively. Cardiac function was evaluated before AMI, 1, 2 and 4 weeks after cell delivery. Hemodynamic parameters were determined 4 weeks after cell infusion. RESULTS: Compared with model group, left ventricular ejection fraction(LVEF), fractional shortening (DeltaFS), left ventricular end-systolic pressure (LVESP) and maximal positive change in filling pressure versus time (+dp/dt max) were improved significantly (all P<0.01), whereas left ventricular end-systolic dimension (LVESD), left ventricular end-diastolic pressure (LVEDP), maximal negative change in filling pressure versus time (-dp/dt max), time constant of left ventricular relaxation (Tc) were lowered in cell infusion groups (all P<0.01). There were no significant differences in cardiac function indexes between intravenous infusion and trans epicardial injection group (all P>0.05). CONCLUSION: Intravenous and trans epicardial delivery of hematopoietic stem cells (HSC) can significantly improve cardiac function, and both methods may be safe and effective for the treatment of AMI. PMID- 18419956 TI - [An experimental study of expression of angiotension converting enzyme 2 in myocardium and effect of telmisartan treatment in pressure-overloaded rats]. AB - OBJECTIVE: To study the effect of hypertension and telmisartan treatment on the protein and gene expression of cardiac angiotensin-converting enzyme 2 (ACE2) in pressure-overloaded rats. METHODS: Coarctation of suprarenal abdominal aorta was reproduced in 8 week-old male Sprague-Dawley (SD) rats and then randomized into 4 groups, including a sham group (n=15), a suprarenal aortic coarctation group (model group, n=12), a suprarenal aortic coarctation with low-dose Telmisartan treatment group (low-dose group, 2 mgxkg(-1)xd(-1), n=11) and a suprarenal aortic coarctation with high-dose Telmisartan treatment group(high-dose group, 10 mgxkg( 1)xd(-1), n=13). Telmisartan or equivalent amount of normal saline was gavaged 24 hours before the operation and once every day afterwards for 3 weeks. At the end of 3 weeks, the concentrations of angiotensin (AngII) in plasma and myocardium were measured by radioimmunoassay. Changes in both protein quantity and gene expressions of both ACE2 and ACE were determined by Western blotting analysis and reverse transcription-polymerase chain reaction (RT-PCR) technique, respectively. RESULTS: Suprarenal abdominal aortic coarctation induced a significant increase in the plasma and myocardium AngII concentration [plasma: (495.1+/-55.6) ng/L vs. (269.2+/-39.5)ng/L, myocardium: (103.6+/-23.7) ng/g vs. (49.5+/-13.5) ng/g, both P<0.01] and expressions of gene and protein of ACE (P<0.01) and ACE2 (P<0.05). Telmisartan further increased the concentration of AngII in plasma and myocardium in a dose-dependent manner [plasma: (702.2+/-40.6) ng/L vs. (612.6+/-35.5) ng/L, myocardium (211.5+/-21.5) ng/g vs. (189.6+/-43.6) ng/g, both P<0.05], and induced a dose-dependent increase in both protein and gene expression of ACE2 (protein 1.16+/-0.06 vs. 0.79+/-0.04, gene 0.54+/-0.08 vs. 0.41+/-0.04, both P<0.05). Expression of ACE2 protein in low-dose and high-dose groups was increased by 1.0 and 1.58 folds respectively, and gene was increased by 1.3 and 1.6 folds (all P<0.05). The expression of ACE protein and gene in model group increased significantly (protein: 2.10+/-1.07 vs. 1.02+/-0.05, gene: 1.93+/-0.09 vs. 0.26+/ 0.09, both P<0.01). Telmisartan had no significant effect on ACE gene and protein expressions (both P>0.05). CONCLUSION: Suprarenal abdominal aortic coarctation induced a significant increases of ACE and ACE2 gene and protein expressions. Telmisartan induces a dose-dependent increases of cardiac ACE2 gene and protein expression,which may be the mechanism of its therapeutic effects. PMID- 18419957 TI - [Protective effects of metallothionein induced by dexamethasone against ischemia/reperfusion injury of myocardium of isolated rat heart]. AB - OBJECTIVE: To investigate the protective effects of metallothionein (MT) induced by dexamethasone (DEX) against ischemia/reperfusion (I/R) injury of myocardium of isolated rat heart. METHODS: Thirty-two Sprague-Dawley (SD) rats were divided randomly into the DEX and control groups. In the former group, the rats were pretreated with DEX, and in latter group distilled water was given before their hearts were isolated for Langendorff perfusion and I/R. MT was assessed by Western blotting. The left ventricular developed pressure (LVDP), maximal change rate of intraventricular pressure rise/down (+/-dp/dt max), coronary artery flow (CF) and reperfusion arrhythmias were observed dynamically before ischemia and during 60-minute reperfusion following 30-minute ischemia, The hearts were perfused with triphenyltetrazolium (TTC) after 60-minute reperfusion. The myocardial infarct size was measured with Adobe Photoshop. The levels of MB isoenzyme of creatine kinase (CK-MB), malonaldehyde (MDA), total superoxide dismutase (T-SOD), CuZn-SOD, catalase (CAT), glutathione peroxidase (GSH-Px) and the activities of Na+-K+-ATPase, Ca2+-Mg2+-ATPase were detected. RESULTS: Compared with control group, the expression of MT was significantly increased (3.085+/-1.065 vs. 1.028+/-0.016, P<0.05), the LVDP, +/-dp/dt max and CF were greatly improved (all P<0.05), the accumulated point of ventricular arrhythmia and the infarct size were significantly reduced in DEX group [(2.00+/-1.41) scores vs. (6.63+/-4.24) scores and (28.38+/-11.22)% vs. (47.39+/-8.30)%, respectively, both P<0.01]. The level of CK-MB was significantly lowered in the DEX group compared with control group [(8.69+/-4.16)U/g vs. (18.15+/-5.59) U/g, P<0.01], and myocardium MDA content was decreased (P<0.05). Moreover, the levels of T-SOD, CuZn-SOD, CAT, GSH-Px, and the activities of Na+-K+-ATPase and Ca2+ Mg2+-ATPase were significantly increased (P<0.05 or/and P<0.01) in DEX group. CONCLUSION: DEX induces upregulation of MT, which attenuates I/R injury in rat heart. PMID- 18419958 TI - [Protective effect of intralipid on myocardial ischemia/reperfusion injury in isolated rat heart]. AB - OBJECTIVE: To investigate whether intralipid could protect perfused hearts of rats against ischemia/reperfusion (I/R) injury when it was administered before (preconditioning) or after the adverse ischemic event (postconditioning), in order to ascertain if intralipid would be a novel therapeutic strategy for myocardial I/R injury. METHODS: Studies were conducted in Langendorff-perfused isolated rat hearts. Twenty-four male Sprague-Dawley (SD) rats were divided into 3 groups randomly. The experimental procedure consisted of balance perfusion for 50 minutes, warm global ischemia (37 centigrade) for 40 minutes and 120 minutes of reperfusion. Preconditioning of hearts in myocardial preconditioning (I-preC group) consisted of 15 minutes of intralipid followed by 15 minutes of wash out before ischemia for 40 minutes and reperfusion for 120 minutes. In myocardial postconditioning (I-postC group) rat hearts were perfused with intralipid for 15 minutes at the onset of reperfusion. Hearts without intralipid treatment served as ischemic control (ISCH) group. Left ventricular mechanical function [heart rate (HR), left ventricular end-diastolic pressure (LVEDP), left ventricular diastolic pressure (LVDP), maximal change rate of intraventricular pressure rise/down (+/-dp/dt max)] were measured during the experiment, cardiac enzyme activity [creatine kinase (CK), lactate dehydrogenase (LDH)] were determined at 20 minutes after balance and 60 minutes after reperfusion. RESULTS: Comparing with ISCH group, the LVDP and +dp/dt max in the I-postC group were significantly higher and LVDEP, -dp/dt max were lower when compared with ISCH groups during reperfusion (all P<0.05). There were no significant differences in above indexes in I-preC group. As compared with the ISCH group, the content of LDH and CK in I preC and I-postC were significantly lower at 60 minutes after reperfusion (all P<0.05). However, there were no significant differences between the I-preC and I postC groups with respect to the levels of LDH and CK. The infarct size (IS) of I preC and I-postC was markedly smaller than that of the ISCH group at 120 minutes of reperfusion [(21.6+/-1.8)%, (15.9+/-1.3)% vs. (46.5+/-3.9)%, both P<0.05]. The IS did not differ between the I-preC and I-postC groups (P>0.05). CONCLUSION: Intralipid administered before or after ischemia can decrease the level of CK, LDH and IS during reperfusion in isolated rat hearts. Intralipid postconditioning improves mechanical function. PMID- 18419959 TI - [Changes in calcitonin gene-related peptide in myocardium and dorsal root ganglion following coronary artery occlusion in rats]. PMID- 18419960 TI - [Relation between back transfusion of blood and inflammatory reaction in off-pump coronary bypass grafting]. PMID- 18419961 TI - [Relationship between Mycoplasma pneumoniae infection and acute myocardial infarction]. PMID- 18419963 TI - [Systems of evaluation scoring of heart function and risks]. PMID- 18419964 TI - [The study of the effect of Cilazapril and valsartan on angiotensin 1 and angiotensin 2 receptors mRNA expression and myocardial interstitial collagen metabolism after myocardial infarction in rats]. PMID- 18419965 TI - [Effects of high-dose glucose-insulin-potassium infusion on preventing occurrence of fatal ventricular arrhythmias in patients with acute myocardial infarction in the early stage]. PMID- 18419966 TI - [Protective effect of ulinastatin against lung injury in patients undergoing cardiopulmonary bypass]. PMID- 18419975 TI - [Surgical treatment of pancreatic neoplasms]. PMID- 18419976 TI - Unicentric Castleman's disease in peripancreatic tissue: case report and review of the literature. AB - Castleman's disease is a rare disorder of the lymphoid tissue that usually occurs in the mediastinum even if extrathoracic involvement, including neck, axilla, mesentery and retroperitoneum, has also been described. We report a case of a 69 years old men with peripancreatic localisation, mimicking a pancreatic neoplasm treated with local excision. Only seven cases of pancreatic and peripancreatic localisation are described in the world-wide literature. This particular site of disease may give troublesome differential diagnosis due to non specific clinical signs and radiological features. Often surgical excision is both diagnostic and therapeutic. PMID- 18419977 TI - [Jejunal perforation from metastatic choriocarcinoma. Case report and review of the literature]. AB - Choriocarcinoma is a rare malignant genital tract tumor, arising in the uterus or in the testis. Primary or metastatic choriocarcinomas of the gastrointestinal tract are infrequent. We report a case of a testis choriocarcinoma presenting as jejunal metastasis with perforation. Histology revealed the origin of this metastatic tumor, allowing us to recognize the primary neoplasm of the testis. A review of literature with PubMed since 1964 and of the references of the papers retrieved was performed. Since 1933 only 30 cases of jejunal choriocarcinomas have been described in literature. In 13 cases jejunal choriocarcinoma presented gastrointestinal hemorrhage, in 4 intestinal intussusception and in 1 case upper abdominal pain and vomiting. Only 5 cases of jejunal perforation have been described. The case presented is the first in literature of jejunal perforation from a metastatic choriocarcinoma of the testis. PMID- 18419978 TI - [Dupuytren's disease simulated by epithelioid sarcoma with atypical perineural invasion of the median nerve. Case report]. AB - Epithelioid sarcoma is an infrequent tumour, especially in upper limb, which nerve spread is not common. We describe the case of a 27 years old caucasian male with initial diagnosis of Dupuytren's disease. He underwent palmar fascia surgery, but the surgical treatment was followed by recurrence of the deformity in retraction of 4th and 5th finger of the left hand. When he presented himself at our attention, the patient underwent surgical biopsy and new histological examination was made with diagnosis of epithelioid sarcoma. We highlight essentially two aspects of this case: the diagnostic delay of tumour caused by simulation of the Dupuytren's disease and the atypical perineural spread along the median nerve. PMID- 18419980 TI - [Two cases of synchronous papillary and medullary thyroid carcinoma]. AB - The Authors describe two cases of simultaneous association of papillary and medullary thyroid carcinoma. They emphasize the treatment of these cases stressing the correlation between the therapeutic criteria and the malignancy of medullary carcinoma. PMID- 18419979 TI - Surgical treatment of differentiated thyroid carcinoma: a retrospective study. AB - INTRODUCTION: We carried out a retrospective analysis of our experience in the management of Differentiated Thyroid Carcinoma (DTC), in order to better define prognostic factors (age, gender, histological type, stage) and outline a standard procedure, where it's possible, for surgical treatment. PATIENTS AND METHODS: Patient population consisted of 432 cases, operated from 1978 to 2003. We carried out 285 operations of total thyroidectomy of which 39 associated to some kind of lymphadenectomy, 66 totalization (21 pts had been operated in other institutes), 60 subtotal thyroidectomies and 21 lobo-isthmectomies. Survival and mortality curves for age, sex, histological type, grading and staging have been calculated. Kaplan-Meyer statistical elaboration for disease-free interval and Mann-Whitney test for the comparison of different clinical and pathological data have been employed. RESULTS: The statistical analysis puts in evidence that on 432 cases examined, with a follow-up from 1 to 25 ys (median = 6.33 ys) and with a drop-out of 60 cases (13.8 %), total mortality for cancer has been of 24 cases (6,4%), with a median interval free by disease of 4.2 ys (range 5 months to 25 ys), and a probability to stay free by disease at 12 and 24 months respectively of 95.1% and 91.6%. The median survival is resulted of 5.8 ys (range 1 to 25 ys) with a probability of survival at 24 and 48 months respectively of 97.5% and 94.3%. The multivariate analysis evidences the most important variables, i.e. age > 45 ys, tumor of intermediate malignancy, with size 1.5 cm, operative M+, significantly condition the prognosis, noticeably getting worse it, independently by the kind of carried out operation. CONCLUSION: Our present therapeutic choices are: 1. total thyroidectomy in the treatment of the apparently benign pathology when bilaterally with spread; the checking at the final histological exam of a cancer makes however think adequate the carried out operation; 2. lobo-isthmectomy in the treatment of unilateral benign pathology or with suspect FNAB for follicular neoplasm; the histological checking of a cancer makes think the operation adequate only in presence of favourable prognostic parameters, but in presence even of just one unfavourable variable, we consider necessary the totalization; 3. total thyroidectomy in presence of a certain or strongly suspected preoperative diagnosis of cancer. PMID- 18419981 TI - [A case of vulvar melanoma: surgical strategy and five years follow-up]. AB - We report a case of a woman with vulvar melanoma, arisen from a plane melanocytic lesion, extended from clitoris to the left minor labium. After diagnostic biopsy, a preoperative lymphoscintigraphy with 99m-Tc nanocolloid was performed. Surgery consisted in a radical vulvectomy, radio-controlled lymphadenectomy and vulvar reconstruction by two bilateral subgluteus soft-tissue flaps, anastomosing the urethral and vaginal stumps. Five years follow-up was negative for neoplastic recurrence. PMID- 18419982 TI - Mayer-Rokitansky-Kuster-Hauser syndrome presenting as vaginal atresia: report of two cases. AB - Mayer-Rokitansky-Kuster-Hauser syndrome (MRKH syndrome) is characterized by Mullerian duct structures agenesis: the vagina atresia is the commonest variant. There can be some anomalies associated, such as renal, skeletal, spine malformations and others. Patients with MRKH can show different presentation from newborn period to adolescence. We report our experience in treatment of the vaginal atresia presenting in two young girls as a sign of MRKH syndrome. PMID- 18419983 TI - [Seroma in residual fibrous capsule after breast implant explantation: a case report]. AB - The Authors present a singular case of seroma which developed in residual fibrous capsule after explantation of breast prostheses without replacement. Mammographic and sonographic findings of the seroma make difficult the diagnosis: mammograms revealed a radiopaque, well-circumscribed mass anterior to the pectoralis major muscle in the region of the prior sub-glandular implant, that was initially misinterpreted as breast implant radiographic feature. It is particularly important review and carefully correlate mammographic and sonographic findings with patient's surgical history for a correct diagnosis. PMID- 18419984 TI - Intestinal obstruction due to idiopathic sclerosing encapsulating peritonitis. Clinical report and review of literature. AB - The sclerosing peritonitis (SP) is a rare illness secondary to the peritoneal dialysis or due to intraperitoneal chemotherapy or the positioning of a peritoneal-jugular shunt in cirrhotic patient with refractory ascites or due to unknown other factors (idiopathic form) like in our patient. The clinical pattern is various and insidious, but when an intestinal occlusive symptomatology is presents an urgent operation is mandatory. The surgical operation is often not easy and asks for a lot of attention especially in the dialyzed subject or in patients with cirrhosis, due to the possibility of postoperative bleeding and other serious complications that can result in fatal outcome. In this report we describe surgical treatment, pathologic pattern and clinical findings of this rare disease. PMID- 18419985 TI - [Major burn: proposal of synthetic prehospital and hospital protocols]. AB - The Authors describe the utility of comprehensive protocols for the treatment of major burns and the adoption of these for the initial and correct management of burns by prehospital and hospital emergency staff. PMID- 18419986 TI - [Retroperitoneal abscess due to asymptomatic duodenal perforation by foreign body. A case report]. AB - We present a case of retroperitoneal abscess due to asymptomatic duodenal perforation by foreign body. Patient has been admitted for lumbar pain and subocclusive crisis, with a medical history negative for acute symptomatology. After both clinical and radiologic evaluation, an abscess-like mass was detected in the context of right psoas muscle. Patient underwent surgical operation and a lumbar abscess has been found containing a foreign body (toothpick). There has been a transduodenal migration of the foreign body, without clinical signs of duodenal perforation. PMID- 18419987 TI - [Diagnostic value of out-patient lymph node biopsy]. AB - Authors review their outpatient lymph node biopsies in order to verify safety and diagnostic accuracy. They didn't observe any complication and in every case, the procedure provided the correct diagnosis. Lymph node biopsy is the procedure of choice to study the pharmacologic therapy non responsive lymph node pathology that is not surely defined by fine needle aspiration, as almost always in lymphomas. PMID- 18419988 TI - Localization of pathological gland's site in primary hyperparathyroidism: ten years experience with MIBI scintigraphy. AB - BACKGROUND: Primary hyperparathyroidism (PHPT) is characterised by pathological hyperfunctioning of one or more of the parathyroid glands leading to excessive parathyroid hormone (PTH) secretion. The aim of this study was to assess the diagnostic capacity of scintigraphy with MIBI, considering the surgical findings and the level of agreement with the result of cervical ultrasonography. PATIENTS AND METHODS: In the period January 1996 to January 2006, 84 cases with PHPT were included in the study, in which scintigraphy with MIBI was used in addition to cervical ultrasonography. All were hospitalised as "short stay surgery" patients and then seen as outpatients at follow-up visits (at 7, 14 and 30 days), during which calcium, phosphorus and PTH values were measured. RESULTS: In 55 cases (65.5%), the site of the pathology was suspected on the basis of the cervical ultrasonography. Scintigraphy with MIBI was positive in 76 cases (90.4%) and negative in the other eight (9.6%). In six of these eight cases the site had been detected by the cervical ultrasonography. In the remaining two cases (2.4%), neither of these examinations gave positive diagnostic findings. In 58 patients we proceeded with the removal of a single adenoma, in 19 cases with the removal of two out of the four glands, and in seven cases with the removal of three out of the four glands, the single formations removed not showing clearly-defined macroscopic characteristics; in two of the seven patients in whom three of the four glands were removed, the preoperative diagnostic examinations had not shown any evidence of gland pathology. CONCLUSIONS: We regard scintigraphy with MIBI as a preoperative diagnostic examination that has modified the surgical approach to PHPT, inclining surgeons towards a mini-invasive surgical procedures. We consider CT, MRI and SPECT techniques to be indicated only in cases of relapse of PHPT, possibly associated with ectopic localisation of the parathyroid gland not identified in the course of previous surgical procedures. PMID- 18419990 TI - Issues in men's health: hormones, sexual function, and quality of life. PMID- 18419991 TI - Recurrence of prostate cancer after radical prostatectomy in a 57-year-old man. PMID- 18419992 TI - Is laparoscopy dying for radical prostatectomy? AB - Training in laparoscopic urology, extensive experimental work, and dedicated surgical performance constitute the foundation of advanced laparoscopic urology, which is currently a reasonable surgical option in pediatrics, reconstructive surgery, and oncology. This article discusses topics related to laparoscopic approaches for radical prostatectomy, focusing on their introduction, development, accomplishments, and current standards and future goals for the minimally invasive treatment of urologic diseases. We highlight the dynamic status of the laparoscopic approach for radical prostatectomy (pure or robotic assisted) that constitutes one of the most rapidly evolving fields in urology. PMID- 18419993 TI - Photodynamic diagnostics of bladder tumors. AB - Approximately 18,000 new cases of urothelial cancer of the bladder occur annually, making it the most frequent urogenital neoplasia representing nearly 4% of all malignomas. Tumor stage defines two subgroups requiring specific therapeutic approaches and implying distinct prognoses. Non-muscle-invasive urothelial cancer recurs frequently and occasionally progresses to muscle invasive stages. Remnant tumor after initial resection is seen as one major factor in high recurrence rates. Although photodynamic diagnostics improving detection and increasing recurrence-free survival has been introduced to broad application, it has yet not been established as standard procedure. This article reviews the current literature and discusses controversial aspects. PMID- 18419994 TI - Update on photoselective vaporization of the prostate. AB - Photoselective vaporization of the prostate (PVP) with a potassium titanyl phosphate (KTP) laser may be the most promising new technology applied to the treatment of benign prostatic hyperplasia (BPH). The specific laser light characteristics and the ideal interactions between KTP lasers and prostatic tissue result in an even and efficient vaporization of the prostate and the formation of a clearly deobstructed prostate cavity. PVP can be a day-care procedure, with few hours of catheterization and minimal postoperative discomfort, offering outcomes at least equivalent to the reference standard transurethral resection of the prostate. Although larger studies are necessary to further define where PVP stands in the management of BPH, this review focuses on contemporary treatment techniques and their limitations, and comments on the outcomes of its current use. PMID- 18419995 TI - Current status of high-intensity focused ultrasound for prostate cancer: technology, clinical outcomes, and future. AB - Based on a review of recently published articles, we evaluated the current status of high-intensity focused ultrasound (HIFU) as a primary treatment option for localized prostate cancer and as a salvage therapy when radiation has failed. With mid- and long-term progression-free survival rates around 70%, negative postoperative prostate biopsies almost 90%, and an excellent morbidity profile, primary HIFU appears to be a valid alternative to active surveillance protocols in low-risk patients and standard therapies in patients with life expectancies of 10 or fewer years. Moreover, HIFU has a considerable potential for local-only recurrence after radiation failure. HIFU is a recent technology, and many improvements will undoubtedly expand its future indications and use for the management of prostate cancer. PMID- 18419996 TI - Updates in endourology. AB - Rapid urologic innovations in minimally invasive treatment are creating exciting new horizons in endourology. However, these new concepts are blurring the traditional boundary between endourology and oncology. Organ-sparing surgery, laparoscopy, robotics systems, and image-guided ablation techniques enable surgeons to develop specifically tailored treatments for patients. We examine recent developments and future prospects for how new technology will continue to advance the field of endourology. PMID- 18419997 TI - Cryoablation and radiofrequency for kidney tumor. AB - The detection of incidental and asymptomatic renal cortical neoplasms has concomitantly increased with radiographic imaging use. The gold standard for treating small renal tumors includes open and laparoscopic partial nephrectomy. Ablative renal procedures intend to duplicate the excellent oncologic outcomes of partial nephrectomy, while decreasing complications and shortening hospitalization time and convalescence. Only short and medium-term data are available, but ablation with cryotherapy or radiofrequency probes may achieve these goals. Ablation also offers the advantages of a minimally invasive surgical approach, with a significantly lower complication rate than partial nephrectomy. Ablated lesions are typically left in situ. Leaving potentially malignant tissues in place, albeit in a nonviable condition, certainly represents a major change in surgical thinking processes. This article reviews the status of cryoablation and radiofrequency ablation, the two ablative modalities currently available for clinical application. PMID- 18419998 TI - Associations among hypospadias, cryptorchidism, anogenital distance, and endocrine disruption. AB - Endocrine disruptors, such as environmental compounds with endocrine-altering properties, may cause hypospadias and cryptorchidism in several species, including humans. Anogenital distance is sexually dimorphic in many mammals, with males having longer anogenital distance on average than females. Animal models of proposed endocrine disruptors have associated prenatal exposure with hypospadias, cryptorchidism, and reduced anogenital distance. Human studies have correlated shorter anogenital distance to in utero exposure to putative endocrine disruptors. We review preliminary data suggesting that anogenital distance is reduced in boys with hypospadia and cryptorchidism. Hence, human hypospadias and cryptorchidism may be associated with reduced anogenital distance as a result of endocrine disruption. PMID- 18419999 TI - Therapy for vesicoureteral reflux: antibiotic prophylaxis, urotherapy, open surgery, endoscopic injection, or observation? AB - Vesicoureteral reflux (VUR) refers to the retrograde flow of urine from the bladder into the ureter and renal pelvis. It generally results from congenital maldevelopment of the ureterovesical junction, although VUR may develop in individuals with abnormally high detrusor pressure. VUR increases a child's susceptibility to pyelonephritis and renal scarring. Treatment goals include the prevention of pyelonephritis, reflux nephropathy, and other complications of reflux. Treatment alternatives include antibiotic prophylaxis, urotherapy (correction of voiding dysfunction), and surgical correction (open, injection therapy, or laparoscopic). Recent studies have challenged the presumed benefit of prophylaxis in children with VUR, while long-term retrospective studies have documented a high rate of hypertension in adults with reflux nephropathy. In addition, the risk of persistent VUR in adulthood is unresolved. These reports have stimulated a reevaluation of the role of various treatment options in children with VUR. PMID- 18420000 TI - Current approaches to the urologic care of children with spina bifida. AB - Most children born with spina bifida, the most common disabling congenital abnormality, have normal renal function. If left untreated, more than half of these children will have serious renal deterioration by age 5. This deterioration is secondary to hostile neurogenic changes of the bladder. Renal development should follow a normal course when close evaluation and intervention are undertaken during the newborn period and toddler years. As children age, attention is directed to quality-of-life issues, such as the establishment of urinary and bowel continence. Teenagers face the responsibility of understanding their medical condition and should begin to assume responsibility for their own care with eventual transition to the adult health care system. This article describes the foundations of management, beginning at birth, for caring for children with spina bifida. PMID- 18420001 TI - The exstrophy-epispadias complex. AB - Patients with the exstrophy-epispadias complex are one of the most challenging groups encountered by pediatric urologists. They generally require surgery involving several reconstruction techniques, usually performed after the first week of life. Common problems in subsequent years include issues related to continence, sexual function, and the appearance of the lower abdomen and genitals. This article reviews major publications over the last few years related to managing patients with this rare, complex, congenital genitourinary anomaly. PMID- 18420002 TI - Urinary tract infections in children. AB - Urinary tract infections can be a significant source of morbidity in the pediatric population. The mainstay of evaluating urinary tract infections in children has been physical examination, urinalysis and culture, and renal and bladder sonography and contrast cystography. However, novel clinical paradigms now consider the importance of various risk factors, such as bacterial virulence and antibiotic-resistance patterns, elimination disorders, and the role of innate immunity and inflammation in determining the likelihood of renal cortical scarring. PMID- 18420003 TI - Disorders of sexual development. AB - In human sexual development, the female phenotype represents the default pathway. Therefore, a failure of testis determination results in the development of the female phenotype, while genetic alterations resulting in partial testicular development can give rise to a wide spectrum of masculinization. In addition to defects in peptide hormones and their receptors, timing of hormonal exposure is also critical to appropriate development. Although much work remains to be done, recent advances in our knowledge have begun to unravel the molecular basis of disorders of sexual development. Consensus statements from investigators have recommended changes in the nomenclature, and further investigations have examined the role of the female and male psyche in patients with these disorders. This review focuses on the diagnosis and management of conditions related to disorders of sexual development. PMID- 18420004 TI - [The STROBE statement or how to improve the reporting of observational studies]. PMID- 18420005 TI - [Economic tools to prioritize patients on the waiting list: discrete choice experiments]. AB - OBJECTIVES: To identify which clinical and social characteristics should be used to prioritize patients on the waiting list for elective surgical procedures. METHODS: A discrete choice experiment (DCE) was conducted using a representative sample of the general population in Navarre (Spain). The sample was selected through simple random sampling by age and sex quotas, stratified by the areas and municipalities of residence of the population aged more than 18 years old. Data were analyzed using Bayesian methods. RESULTS: The relative weights of attributes show that the most important attributes when prioritizing patients were the disease, the cost of the intervention, and waiting time. As expected, severity of illness was the most important attribute and, contrary to prior expectations, improvements in health were considered less important. These findings show that prioritization according to waiting time alone may not take into account other issues considered important by the general public. CONCLUSIONS: Patients should not be prioritized according to waiting time only. An interesting finding that should be analyzed in future is that cost was considered an important prioritization criterion. This study provides a further example of the potential of DCE in health economics. If its limitations are borne in mind, this tool may be useful to develop prioritization scoring systems for patients on waiting lists. PMID- 18420006 TI - [Childhood obesity: pediatricians' attitudes and opinions]. AB - OBJECTIVES: To identify the Spanish pediatricians' opinions and attitudes towards obesity, in relation with its treatment, prevention, and care organization, barriers they find in its treatment, and to know the tools they consider more useful to have for the management of obese children. METHODS: Mail survey posted to the primary care pediatricians of Castilla-Leon (Spain) exploring: willingness to act against obesity, utility of therapies and preventive strategies, barriers found in treating obese patients, most needed tools for treating obesity, autoefficacy, and support to an obese children management programme in primary care. RESULTS: There was a broad consensus in seeing obesity as an important health problem that demands action from pediatricians. This willingness to action decreased when bigger personal barriers were found. The most frequently encountered barriers came from the social milieu: easy access and advertisement of certain foods and beverages, lack of implication of parents, lack of perception of a weight problem in children and parents. Training was the most trusted tool. Pediatricians considered themselves as some or low efficacious in treating obesity. The effectiveness of an obese children management program in primary care was surpassed by the work it would need. CONCLUSIONS: In spite of their high willingness to act against obesity, pediatricians feel themselves limited because they find barriers mainly from the social and cultural milieus. They ask for training to fight pediatric obesity. PMID- 18420007 TI - [Impact of new therapeutic products for the treatment of chronic obstructive pulmonary disease and asthma in primary care in Madrid between 1996 and 2005]. AB - OBJECTIVES: To analyze the trend of use of new drugs for the treatment of chronic obstructive pulmonary disease in primary care of Madrid (Spain) between 1996 and 2005, and to evaluate its impact in terms of supply, use and costs. METHODS: Drug utilization study of new products of the R03 group (Anatomical Therapeutic Chemical Classificaction System: drugs for obstructive airway diseases) prescribed by doctors of primary care of Madrid, during 1996 and 2005. Information on drug utilization and cost was obtained from the prescription database. Date was expressed in daily doses defined by 1,000 inhabitants and day. RESULTS: The supply experiences few quantitative variations although of great qualitative interest, when disappearing drugs without therapeutic utility and commercializing 6 therapeutic new products. In 2005, the use of the 6 new products generates 28.03% of the total of the consumption and 79% of the costs. The new drugs of greater impact are salmeterol/fluticasone and tiotropio. CONCLUSIONS: The supply of medicines qualitatively improves when disappearing drugs without therapeutic utility. The consumption of therapeutic new products has a great impact on the total consumption of the R03 group and, mainly, in the costs by its elevated price, in spite of contributing only limited therapeutic advantages. The new drugs have been gotten up at great speed to the prescription. PMID- 18420008 TI - [Impact of heat waves on daily mortality in distinct age groups]. AB - OBJECTIVES: To establish whether the daily maximum temperature during heat waves beyond which daily mortality increases differs in distinct age groups and to quantify the mortality increment by these age groups. METHODS: The dependent variable was daily mortality from all causes except for accidents in the city of Madrid from 1986 to 1997. The age groups analyzed were the following: 0-10 years, 18-44 years, 45-64 years, 65-74 years, and older than 75 years. Data on the daily maximum temperature in the summer months was used. Trend and seasonalities were adequately controlled for, as were air pollutant concentrations. Autoregressive moving average (ARIMA) and Poisson regression models were used. RESULTS: In all age groups except that aged less than 10 years old, there was an association between extremely high temperatures and mortality. Independently of age group, the temperature threshold above which mortality increased coincided with the 95th percentile of the maximum temperature data series for the summer months, which in Madrid is 36.5 degrees C. The daily mortality increment for each degree above this threshold was practically constant between the ages of 18 to 64 years but markedly increased in persons aged more than 65 years old. CONCLUSIONS: Mortality is associated with high temperatures in people younger than 64 years old and therefore prevention plans should not be focused exclusively on the elderly. PMID- 18420009 TI - [Factors related to refuse antiretroviral therapy in prison]. AB - INTRODUCTION: Despite the benefits of highly-active antiretroviral therapy (HAART) on morbidity and mortality, some seropositive patients refuse to accept this treatment. Prisons provide easier access to this population. OBJECTIVE: To determine the psychosocial characteristics of prisoners who refuse HAART. METHODS: We performed a cross-sectional study in 580 seropositive prisoners in 3 hospitals in Andalusia (Spain). The dependent variable was being under treatment or refusal to be so. The independent variables were sociodemographic and psychosocial factors related to the prison environment and clinical factors related to health status and drug addiction. A logistic regression analysis was performed to determine which factors were related with refusal to accept HAART. RESULTS: HAART was recommended to 73.1% of seropositive prisoners. This treatment was refused by 23.1% of these prisoners and was accepted by 76.9%. The factors related to refusal to accept HAART were high viral load, worse self-perceived health status, a greater number of stays in prison, and being visited by persons other than relatives. CONCLUSIONS: There is a group of prisoners with specific characteristics that refuse HAART. Specific interventions should be performed in these prisoners to make them aware of the effects of their decision on the course of their disease. PMID- 18420010 TI - Access to sterile syringes among young drug injectors in Madrid and Barcelona and its association with risk behaviour. AB - BACKGROUND AND OBJECTIVE: To evaluate access to sterile syringes and its association with injection risk behaviour in Madrid and Barcelona. MATERIALS AND METHODS: Cross-sectional community study by computer-assisted personal interview in 465 young heroin injectors between 2001 and 2003. RESULTS: Some 4.2% had not obtained any free sterile syringes in the previous 12 months. In Madrid 32.1% had obtained all their sterile syringes free of charge (ASSF), versus 44.6% in Barcelona (p<0.01). Not sharing (not using syringes used by someone else and not front/backloading) was associated with obtaining ASSF (OR=1.69) and with sporadic injection (OR=1.83). Not reusing one's own syringes was associated with the same two variables (OR=4.02 and OR=2.50, respectively). CONCLUSIONS: Access to sterile syringes is very high in Madrid and Barcelona, although the two cities have different approaches. The acquisition of all syringes free of charge should be facilitated, especially among frequent injectors. PMID- 18420011 TI - [Utilization of dermatology services among the immigrant population]. AB - OBJECTIVE: To determine the utilization rate of a dermatology service among the immigrant population and compare this rate with that among the autochthonous population. METHODS: Over a 1-year period, data on all patients attended in the Dermatology Service of Health Department 19 in the autonomous community of Valencia were prospectively collected. RESULTS: Of 39,160 consultations, 1,625 were made by immigrants (4.1%). Attendance for dermatologic emergencies was greater in immigrants than in the autochthonous population (7.7% vs 3.0%; p<0.001), while surgical activity was lower in immigrants than in Spanish-born patients (2.6% vs 5%; p<0.001). Demand for consultation in the immigrant population was 5.7 visits per 100 immigrants, lower than that in the autochthonous population (16.2). The standardized rate ratio was 0.34. North Africans (5.6 per 100 immigrants), East Europeans (4.8), sub-Saharan Africans (5.3) and Asians (4.2) consulted less than Latin Americans (8.5; p<0.001). CONCLUSIONS: Demand was lower in the immigrant than in the autochthonous population, and the utilization pattern differed according to country. PMID- 18420012 TI - [Patients with multimorbidity in the hospital setting]. AB - OBJECTIVES: Hospitals attend patients with multiple chronic diseases. The aim of the present study was to estimate the prevalence of these patients, as well as their main sociodemographic and management characteristics. METHODS: We performed a descriptive study based on information from the minimum data set and a functional definition of patients with multimorbidity. Age, sex, admitting specialty, cause of admission, discharge and days of hospital stay were estimated for patients discharged from a university hospital in 2003. RESULTS: Patients with multimorbidity represented 16.9% (95% CI: 15.8-18.1%) of admissions. These patients were mainly elderly men who tended to be admitted urgently to medical specialties. Discharge to home was less frequent and days of hospital stay tended to be longer. CONCLUSIONS: Patients with multiple comorbidities have well-defined characteristics that allow them to be retrospectively identified through analysis of the minimum data set. Although nearly all services discharged these patients, most of them were discharged from medical specialties. PMID- 18420013 TI - [Comment. Healthcare for patients with multiple disorders. Are there opportunities for improvement?]. PMID- 18420015 TI - [Discrete-event simulation models in the economic evaluation of health technologies and health products]. AB - The use of mathematical models to assess therapeutic alternatives is increasing in the economic evaluation of health technologies and services and these models are becoming an increasingly important aid to decision making in health care. Until now, 2 types of model have been used, depending to some extent on the disease to be studied: decision trees have been used for acute diseases and Markov models in chronic or recurrent diseases. However, both models present major limitations when addressing complex processes or diseases. Consequently, interest in, and the use of, discrete-event simulation is growing. The present article aims to describe the main characteristics of discrete-event simulation, the state of the art in this field, and the advantages of these models with respect to other kinds of models in health economics, especially in the evaluation of health technologies and product assessment. PMID- 18420014 TI - [The Strengthening the Reporting of Observational Studies in Epidemiology [STROBE] statement: guidelines for reporting observational studies]. AB - Much biomedical research is observational. The reporting of such research is often inadequate, which hampers the assessment of its strengths and weaknesses and of a study's generalisability. The Strengthening the Reporting of Observational Studies in Epidemiology (STROBE) initiative developed recommendations on what should be included in an accurate and complete report of an observational study. We defined the scope of the recommendations to cover three main study designs: cohort, case-control, and cross-sectional studies. We convened a 2-day workshop in September, 2004, with methodologists, researchers, and journal editors to draft a che-cklist of items. This list was subsequently revised during several meetings of the coordinating group and in e-mail discussions with the larger group of STROBE contributors, taking into account empirical evidence and methodological considerations. The workshop and the subsequent iterative process of consultation and revision resulted in a checklist of 22 items (the STROBE statement) that relate to the title, abstract, introduction, methods, results, and discussion sections of articles. 18 items are common to all three study designs and four are specific for cohort, case-control, or cross-sectional studies. A detailed explanation and elaboration document is published separately and is freely available on the websites of PLoS Medicine, Annals of Internal Medicine, and Epidemiology. We hope that the STROBE statement will contribute to improving the quality of reporting of observational studies. PMID- 18420016 TI - [Real time information. A necessary tool in epidemiological surveillance]. AB - Since 2004 an electronic epidemiological surveillance system (AVE) has been developed and implemented for infectious diseases in the autonomous region of Valencia that allows data collection from notifiable diseases in real time concerning outbreaks and alerts, as well as analysis and dissemination of the information. Within the autonomous region of Valencia, the system is used by 17 epidemiology units, which act as the first level of specialized surveillance. The electronic surveillance system is used by all these units, as well as by physicians working in primary and specialized care, who can introduce information from the ambulatory information system while the patient is in the practice room. The system is able to capture the demographic data from the patient and the physician can add any other information that is considered important for the case detected. The system receives new laboratory test results overnight from the Microbiological Surveillance Network (RedMIVA). AVE can record outbreaks of notifiable diseases as well as any unusual clusters or changing patterns of any disease. The department of epidemiology has access to the whole system for analysis of the information and for quality control of the epidemiological surveillance. The system provides timely and comprehensive information to facilitate public health action in individual cases of infectious disease. PMID- 18420018 TI - [Anonymous peer review? No way!]. PMID- 18420019 TI - [Mirrors and mirages of a reality out of focus]. PMID- 18420020 TI - Homogeneous and label-free fluorescence detection of single-nucleotide polymorphism using target-primed branched rolling circle amplification. AB - We present a simple, sensitive, and cost-effective fluorescent assay of single nucleotide polymorphism (SNP) with target-primed branched rolling circle amplification (TPBRCA). Designed padlock probe is circularized after perfect hybridization to mutant DNA. Then rolling circle amplification (RCA) reaction can be initiated from the mutant DNA that acts as primer and generates a long tandem single-stranded DNA (ssDNA) product. At the same time, the introduction of a reverse primer complementary to the target-primed RCA products leads to the branched RCA and eventually generates the various lengths of ssDNA and double stranded DNA products, which are sensitively detected using SYBR Green I (SG) fluorescence dye. In contrast, the wild DNA contains a single mismatched base with the padlock probe and primes only a limited extension with the unligated padlock probe, generating weak background fluorescence with the addition of SG. Due to the excellent specificity and powerful amplification of TPBRCA reaction, the mutant DNA was distinctively differentiated from the wild DNA in a homogeneous and label-free manner. The assay is sensitive and specific enough to detect 5-amol (8.6-fM) mutant DNA strands. It was possible to accurately determine the mutant allele frequency as low as 1.0%. PMID- 18420021 TI - Error minimization algorithm for comparative quantitative PCR analysis: Q-Anal. AB - Current methods for comparative quantitative polymerase chain reaction (qPCR) analysis, the threshold and extrapolation methods, either make assumptions about PCR efficiency that require an arbitrary threshold selection process or extrapolate to estimate relative levels of messenger RNA (mRNA) transcripts. Here we describe an algorithm, Q-Anal, that blends elements from current methods to by pass assumptions regarding PCR efficiency and improve the threshold selection process to minimize error in comparative qPCR analysis. This algorithm uses iterative linear regression to identify the exponential phase for both target and reference amplicons and then selects, by minimizing linear regression error, a fluorescence threshold where efficiencies for both amplicons have been defined. From this defined fluorescence threshold, cycle time (Ct) and the error for both amplicons are calculated and used to determine the expression ratio. Ratios in complementary DNA (cDNA) dilution assays from qPCR data were analyzed by the Q Anal method and compared with the threshold method and an extrapolation method. Dilution ratios determined by the Q-Anal and threshold methods were 86 to 118% of the expected cDNA ratios, but relative errors for the Q-Anal method were 4 to 10% in comparison with 4 to 34% for the threshold method. In contrast, ratios determined by an extrapolation method were 32 to 242% of the expected cDNA ratios, with relative errors of 67 to 193%. Q-Anal will be a valuable and quick method for minimizing error in comparative qPCR analysis. PMID- 18420022 TI - Characterization of photosystem II in salt-stressed cyanobacterial Spirulina platensis cells. AB - PSII activity was inhibited after Spirulina platensis cells were incubated with different salt concentrations (0-0.8 M NaCl) for 12 h. Flash-induced fluorescence kinetics showed that in the absence of DCMU, the half time of the fast and slow components decreased while that of the middle component increased considerably with increasing salt concentration. In the presence of DCMU, fluorescence relaxation was dominated by a 0.6s component in control cells. After salt stress, this was partially replaced by a faster new component with half time of 20-50 ms. Thermoluminescence measurements revealed that S(2)Q(A)(-) and S(2)Q(B)(-) recombinations were shifted to higher temperatures in parallel and the intensities of the thermoluminescence emissions were significantly reduced in salt-stressed cells. The period-four oscillation of the thermoluminescence B band was highly damped. There were no significant changes in contents of CP47, CP43, cytochrome c550, and D1 proteins. However, content of the PsbO protein in thylakoid fraction decreased but increased significantly in soluble fraction. The results suggest that salt stress leads to a modification of the Q(B) niche at the acceptor side and an increase in the stability of the S(2) state at the donor side, which is associated with a dissociation of the PsbO protein. PMID- 18420023 TI - Oxidative stress and mitochondrial dysfunction in neurodegeneration; cardiolipin a critical target? AB - Oxidative stress and subsequent impairment of mitochondrial function is implicated in the neurodegenerative process and hence in diseases such as Parkinson's and Alzheimer's disease. Within the brain, neuronal and astroglial cells can display a differential susceptibility to oxidant exposure. Thus, astrocytes can up regulate glutathione availability and, in response to mitochondrial damage, glycolytic flux. Whilst neuronal cells do not appear to possess such mechanisms, neuronal glutathione status may be enhanced due to the trafficking of glutathione precursors from the astrocyte. However, when antioxidants reserves are not sufficient or the degree of oxidative stress is particularly great, mitochondrial damage occurs, particularly at the level of complex IV (cytochrome oxidase). Whilst the exact mechanism for the loss of activity of this enzyme complex is not know, it is possible that loss and/or oxidative modification of the phospholipid, cardiolipin is a critical factor. Consequently, in this short article, we also consider (a) cardiolipin metabolism and function, (b) the susceptibility of this molecule to undergo oxidative modification following exposure to oxidants such as peroxynitrite, (c) loss of mitochondrial cardiolipin in neurodegenerative disorders, (d) methods of detecting cardiolipin and (e) possible therapeutic strategies that may protect cardiolipin from oxidative degradation. PMID- 18420024 TI - Ultrafast ligand binding dynamics in the active site of native bacterial nitric oxide reductase. AB - The active site of nitric oxide reductase from Paracoccus denitrificans contains heme and non-heme iron and is evolutionarily related to heme-copper oxidases. The CO and NO dynamics in the active site were investigated using ultrafast transient absorption spectroscopy. We find that, upon photodissociation from the active site heme, 20% of the CO rebinds in 170 ps, suggesting that not all the CO transiently binds to the non-heme iron. The remaining 80% does not rebind within 4 ns and likely migrates out of the active site without transient binding to the non-heme iron. Rebinding of NO to ferrous heme takes place in approximately 13 ps. Our results reveal that heme-ligand recombination in this enzyme is considerably faster than in heme-copper oxidases and are consistent with a more confined configuration of the active site. PMID- 18420025 TI - Secondary and tertiary structures of the transmembrane domains of the translocator protein TSPO determined by NMR. Stabilization of the TSPO tertiary fold upon ligand binding. AB - Numerous biological functions are attributed to the peripheral-type benzodiazepine receptor (PBR) recently renamed translocator protein (TSPO). The best characterized function is the translocation of cholesterol from the outer to inner mitochondrial membrane, which is a rate-determining step in steroid biosynthesis. TSPO drug ligands have been shown to stimulate pregnenolone formation by inducing TSPO-mediated translocation of cholesterol. Until recently, no direct structural data on this membrane protein was available. In a previous paper, we showed that a part of the mouse TSPO (mTSPO) C-terminal region adopts a helical conformation, the side-chain distribution of which provides a groove able to fit a cholesterol molecule. We report here on the overall structural properties of mTSPO. This study was first undertaken by dissecting the protein sequence and studying the conformation of five peptides encompassing the five putative transmembrane domains from (1)H-NMR data. The secondary structure of the recombinant protein in micelles was then studied using CD spectroscopy. In parallel, the stability of its tertiary fold was probed using (1)H-(15)N NMR. This study provides the first experimental evidence for a five-helix fold of mTSPO and shows that the helical conformation of each transmembrane domain is mainly formed through local short-range interactions. Our data show that, in micelles, mTSPO exhibits helix content close to what is expected but an unstable tertiary fold. They reveal that the binding of a drug ligand that stimulates cholesterol translocation is able to stabilize the mTSPO tertiary structure. PMID- 18420026 TI - Nectin-like molecule 1 is a glycoprotein with a single N-glycosylation site at N290KS which influences its adhesion activity. AB - Nectin-like molecule 1 (NECL1)/CADM3/IGSF4B/TSLL1/SynCAM3, from now on referred to as NECL1, is a neural tissue-specific immunoglobulin-like cell-cell adhesion molecule which has Ca(2+)-independent homo- or heterophilic cell-cell adhesion activity and plays an important role in the formation of synapses, axon bundles and myelinated axons. Here we first detected the expression of NECL1 in human fetal and adult brains, and mouse brains at different developmental stages. The results indicated that two bands with molecular weights of about 62 kDa and 48 kDa were found in human fetal brain, while only one band with a molecular weight of about 48 kDa was found in human adult brain; two bands with molecular weights of about 62 kDa and 48 kDa whose expression level gradually increased were also found from mouse E16 to P14, while only one band with a molecular weight of about 48 kDa was found from P14. Bioinformatics analysis showed there were two putative N-glycosylation sites within human NECL1 at positions N25LS and N290KS and within mouse Necl1 at positions N23LS and N288KS, respectively. There was no O glycosylation site in either human NECL1 or mouse Necl1. Based on the results of N-Glycosidase F treatment with human fetal brain tissue and lysates from transient transfection with human wild-type or glycosylation site mutant NECL1 in 293ET cells, we demonstrated that human NECL1 is an N-linked glycoprotein with a single glycosylation site at position N290KS. Cell aggregation assay further showed there was an increased adhesion activity after the glycosylation site mutation of NECL1 molecule. PMID- 18420027 TI - Apoptosis induced by endoplasmic reticulum stress involved in diabetic kidney disease. AB - Endoplasmic reticulum stress has been suggested to play a crucial role in the pathogenesis of diabetic complications. However, whether it is involved in the renal injury of diabetic nephropathy is still not known. We investigated the involvement of ER-associated apoptosis in kidney disease of streptozocin (STZ) induced diabetic rats. We used albuminuria examination, hematoxylin & eosin (H&E) staining and TUNEL analysis to identify the existence of diabetic nephropathy and enhanced apoptosis. We performed immunohistochemistry, Western blot, and real time PCR to analyze indicators of ER molecule chaperone and ER-associated apoptosis. GRP78, the ER chaperone, was up-regulated significantly in diabetic kidney compared to control. Furthermore, three hallmarks of ER-associated apoptosis, C/EBP homologous protein (CHOP), c-JUN NH2-terminal kinase (JNK) and caspase-12, were found to have activated in the diabetic kidney. Taken together, those results suggested that apoptosis induced by ER stress occurred in diabetic kidney, which may contribute to the development of diabetic nephropathy. PMID- 18420028 TI - Transplantation of bone marrow derived cells promotes pancreatic islet repair in diabetic mice. AB - The transplantation of bone marrow (BM) derived cells to initiate pancreatic regeneration is an attractive but as-yet unrealized strategy. Presently, BM derived cells from green fluorescent protein transgenic mice were transplanted into diabetic mice. Repair of diabetic islets was evidenced by reduction of hyperglycemia, increase in number of islets, and altered pancreatic histology. Cells in the pancreata of recipient mice co-expressed BrdU and insulin. Double staining revealed beta cells were in the process of proliferation. BrdU(+) insulin(-) PDX-1(+) cells, Ngn3(+) cells and insulin(+) glucagon(+) cells, which showed stem cells, were also found during beta-cell regeneration. The majority of transplanted cells were mobilized to the islet and ductal regions. In recipient pancreas, transplanted cells simultaneously expressed CD34 but did not express insulin, PDX-1, Ngn3, Nkx2.2, Nkx6.1, Pax4, Pax6, and CD45. It is concluded that BM derived cells especially CD34(+) cells can promote repair of pancreatic islets. Moreover, both proliferation of beta cells and differentiation of pancreatic stem cells contribute to the regeneration of beta cells. PMID- 18420029 TI - A mis-expression study of factors affecting Drosophila PNS cell identity. AB - Drosophila PNS sense organs arise from single sensory organ precursor (SOP) cells through a series of asymmetric divisions. In a mis-expression screen for factors affecting PNS development, we identified string and dappled as being important for the proper formation of adult external sensory (ES) organs. string is a G2 regulator. dappled has no described function but is implicated in tumorigenesis. The mis-expression effect from string was analysed using timed over expression during adult ES-organ and, for comparison, embryonic Chordotonal (Ch) organ formation. Surprisingly, string mis-expression prior to SOP division gave the greatest effect in both systems. In adult ES-organs, this lead to cell fate transformations producing structural cells, whilst in the embryo organs were lost, hence differences within the lineages exist. Mis-expression of dappled, lead to loss and duplications of entire organs in both systems, potentially affecting SOP specification, in addition to affecting neuronal guidance. PMID- 18420030 TI - Selection of diagnostic tests for clinical decision making and translation to a problem oriented medical record. AB - The leading function of the physician is the clinical reasoning, which involves appropriate investigation of the problems of the patient, formulation of a diagnostic suspect based on the patient's symptoms and signs, gathering of additional relevant information, to select necessary tests and administration of the most suitable therapy. The problems of the patient are expressed by symptoms or signs or abnormal test results, requested for a variety of reasons. The entire scientific, as well as diagnostic approach, is based on three steps: to stumble in a problem; to try a solution through a hypothesis; to disprove or to prove the hypothesis by a process of criticism. Clinicians use the information obtained from the history and physical examination to estimate initial (or pre-test) probability and then use the results from tests and other diagnostic procedures to modify this probability until the post-test probability is such that the suspected diagnosis is either confirmed or ruled out. When the pre-test probability of disease is high, tests characterized by high specificity will be preferred, in order to confirm the diagnostic suspect. When the pre-test probability of disease is low, a test with high sensitivity is advisable to exclude the hypothetical disease. The above mentioned process of decision making has been transferred to a problem oriented medical record that is currently employed in our Clinic. PMID- 18420031 TI - Improving training in laboratory medicine. AB - Recent advances in medicine and technology, combined with an ever-growing workload, have increased the demand for skilled professionals in laboratory medicine. The specific need for trained physicians and scientists raises important questions about the content of training programs and about the ideal characteristics of the human products of the training programs. Excellent recent publications in several countries have addressed the scientific and technical components of training and the competencies that trainees are expected to develop. These publications will provide important guidance for training programs and for trainees for the foreseeable future. An additional goal of training is to produce members of a profession. These professionals will 1) aspire to meeting the challenges of the field with creativity, 2) be involved with the profession and the world and 3) function in a manner informed by their thorough grounding in professional and medical ethics. In the promising future of laboratory medicine, a focus on professional aspects is essential to meeting the potential of the field to contribute to health of the patients we serve. PMID- 18420032 TI - Models of learning, training and progress evaluation of medical students. AB - Medical education has changed during the last century. Teachercentred medical education has been replaced by a studentcentred one. Some general educational principles have been adopted in medicine faculties, leading to more active students involved in learning, and interactive methods have been introduced, and the role of experience has been focused. Disciplinebased medical education is inadequate for developing comprehensive competence and doctoring. Integrated multidisciplinary learning, together with new clerkship approaches and organization, represent the future of medical education. Continuing, comprehensive, assessment can measure progression in knowledge, skills, attitudes and values. Progress test have recently been introduced in some medical school as a reliable tool for assessing the progressive performance of students in time and monitoring the entire learning process. PMID- 18420033 TI - The EC4 European Register of Specialists in Clinical Chemistry and Laboratory Medicine. AB - The European Register of Specialists in Clinical Chemistry and Laboratory Medicine was established by the European Communities Confederation of Clinical Chemistry and Laboratory Medicine (EC4) in 1997. The main aims of the Register are to ensure a high quality of professional standards and practice in the European Union (EU) countries, and to assist free movement of professionals within the EU, in accordance with European Directives. We report here the latest developments on the structure of the register, a description of the application process and data about the number of registered members, and the relationship between the Register and the European Commission Directive of Professional Qualifications. PMID- 18420034 TI - Detection of lung cancer using weighted digital analysis of breath biomarkers. AB - BACKGROUND: A combination of biomarkers in a multivariate model may predict disease with greater accuracy than a single biomarker employed alone. We developed a non-linear method of multivariate analysis, weighted digital analysis (WDA), and evaluated its ability to predict lung cancer employing volatile biomarkers in the breath. METHODS: WDA generates a discriminant function to predict membership in disease vs no disease groups by determining weight, a cutoff value, and a sign for each predictor variable employed in the model. The weight of each predictor variable was the area under the curve (AUC) of the receiver operating characteristic (ROC) curve minus a fixed offset of 0.55, where the AUC was obtained by employing that predictor variable alone, as the sole marker of disease. The sign (+/-) was used to invert the predictor variable if a lower value indicated a higher probability of disease. When employed to predict the presence of a disease in a particular patient, the discriminant function was determined as the sum of the weights of all predictor variables that exceeded their cutoff values. The algorithm that generates the discriminant function is deterministic because parameters are calculated from each individual predictor variable without any optimization or adjustment. We employed WDA to re-evaluate data from a recent study of breath biomarkers of lung cancer, comprising the volatile organic compounds (VOCs) in the alveolar breath of 193 subjects with primary lung cancer and 211 controls with a negative chest CT. RESULTS: The WDA discriminant function accurately identified patients with lung cancer in a model employing 30 breath VOCs (ROC curve AUC=0.90; sensitivity=84.5%, specificity=81.0%). These results were superior to multilinear regression analysis of the same data set (AUC=0.74, sensitivity=68.4, specificity=73.5%). WDA test accuracy did not vary appreciably with TNM (tumor, node, metastasis) stage of disease, and results were not affected by tobacco smoking (ROC curve AUC=0.92 in current smokers, 0.90 in former smokers). WDA was a robust predictor of lung cancer: random removal of 1/3 of the VOCs did not reduce the AUC of the ROC curve by >10% (99.7% CI). CONCLUSIONS: A test employing WDA of breath VOCs predicted lung cancer with accuracy similar to chest computed tomography. The algorithm identified dependencies that were not apparent with traditional linear methods. WDA appears to provide a useful new technique for non-linear multivariate analysis of data. PMID- 18420035 TI - Prognostic significance of BP1 mRNA expression level in patients with non-small cell lung cancer. AB - OBJECTIVES: To examine the association of BP1 mRNA level with tumor characteristics and clinical prognosis in non-small cell lung cancer (NSCLC) patients. DESIGN AND METHODS: Tumor specimens from 98 NSCLC patients who underwent surgical resection were quantitatively determined for BP1 mRNA expression by real-time RT-PCR. RESULTS: BP1 mRNA was expressed at significantly higher levels in tumors than in adjacent nontumorous tissues and normal lung samples. The level of BP1 transcript was significantly associated with tumor histological type and cell differentiation grade, but not related with other clinicopathological factors and p53 mutations. Patients with high BP1 mRNA expression had a poorer prognosis in terms of both disease-free survival (DFS) and overall survival (OS) rates. Additionally, BP1 mRNA expression level was an independent prognostic factor for DFS. CONCLUSIONS: BP1 may be part of a pathway contributing to NSCLC development and/or progression. BP1 mRNA level could be a novel prognostic marker for NSCLC. PMID- 18420036 TI - Proteins in the insulin-secreting cell line MIN6 bind the imidazoline compound BL11282. AB - The imidazoline BL11282 stimulates insulin release and alters islet proteomes. Subcellular fractions of MIN6 cells showed that the membrane fraction exhibited binding to BL11282 on a Biacore chip and to BL11282-labelled magnetic beads. Bound material extracted from the beads showed a approximately 50 kDa differential band upon SDS-PAGE and a weaker 100 kDa band. The former was sensitive to competitive removal by preincubation of the fraction with BL11282, then highlighting the approximately 100 kDa band. Masspectrometric analysis revealed the approximately 50 kDa band to be EF1A and the approximately 100 kDa band to be glucose regulated P94, both of interest in insulin synthesis and secretion. PMID- 18420037 TI - Actin in the endocytic pathway: from yeast to mammals. AB - Genetic analysis of endocytosis in yeast early pointed to the essential role of actin in the uptake step. Efforts to identify the machinery involved demonstrated the important contribution of Arp2/3 and the myosins-I. Analysis of the process using live-cell fluorescence microscopy and electron microscopy have recently contributed to refine molecular models explaining clathrin and actin-dependent endocytic uptake. Increasing evidence now also indicates that actin plays important roles in post-internalization events along the endocytic pathway in yeast, including transport of vesicles, motility of endosomes and vacuole fusion. This review describes the present knowledge state on the roles of actin in endocytosis in yeast and points to similarities and differences with analogous processes in mammals. PMID- 18420038 TI - Targeting DNA quadruplexes with distamycin A and its derivatives: an ITC and NMR study. AB - The use of small molecules that bind and stabilize G-quadruplex structures is emerging as a promising way to inhibit telomerase activity in tumor cells. In this paper, isothermal titration calorimetry (ITC) and 1H NMR studies have been conducted to examine the binding of distamycin A and its two carbamoyl derivatives (compounds 1 and 2) to the target [d(TGGGGT)]4 and d[AG3(T2AG3)3] quadruplexes from the Tetrahymena and human telomeres, respectively. The interactions were examined using two different buffered solutions containing either K+ or Na+ at a fixed ionic strength, to evaluate any influence of the ions present in solution on the binding behaviour. Experiments reveal that distamycin A and compound 1 bind the investigated quadruplexes in both solution conditions; conversely, compound 2 appears to have a poor affinity in any case. Moreover, these studies indicate that the presence of different cations in solution affects the stoichiometry and thermodynamics of the interactions. PMID- 18420039 TI - Characterization of the 5' region of the Leishmania infantum LORIEN/MAT2 gene cluster and role of LORIEN flanking regions in post-transcriptional regulation. AB - LORIEN (encoding a protein that contains a SP-RING/Miz zinc-finger motif present in a group of proteins involved in the Small Ubiquitin-related Modifier -SUMO- conjugation pathway) and MAT2 (encoding the methionine adenosyltransferase -MAT-) genes are arranged as two alternating copies in a head-to-tail configuration, with the LORIEN gene as the first copy of the cluster. The 5880bp preceding the first LORIEN gene copy were compared to the same region of L. major, showing a 93% identity between them. Bioinformatic analysis of this region predicted the presence of a 747-bp ORF encoding a hypothetical protein of 248 amino acids. Transcription of this ORF was confirmed by run-on assays and RT-PCR. Expression of the LORIEN gene was tested in both the promastigote and amastigote stages. Transcription arrest evidenced that LORIEN mRNA stability was very similar in both stages of the parasite life cycle. Protein synthesis inhibition by cycloheximide led to an increase in the steady-state levels of LORIEN transcripts only during the promastigote stage, pointing out to the existence of different stage-dependent mechanisms operating on the post-transcriptional regulation of this gene. The role of the LORIEN untranslated regions (5'UTR and 3'UTR) in post transcriptional regulation was analysed using the luciferase (luc) reporter gene. Results evidenced that the 5'UTR was responsible for a low reporter gene expression, whereas the intergenic region (IR) between LORIEN and MAT2 genes provided high luc levels. However, the 3'UTR seemed to lack regulatory elements. Basing on these results, a model of regulation for the LORIEN gene is proposed. PMID- 18420040 TI - Gene regulatory networks that control the specification of neural-crest cells in the lamprey. AB - The lamprey is the only basal vertebrate in which large-scale gene perturbation analyses are feasible at present. Studies on this unique animal model promise to contribute both to the understanding of the basic neural-crest gene regulatory network architecture, and evolution of the neural crest. In this review, we summarize the currently known regulatory relationships underlying formation of the vertebrate neural crest, and discuss new ways of addressing the many remaining questions using lamprey as an experimental model. PMID- 18420041 TI - Speciation of inorganic platinum-chloride complexes in spiked environmental samples by SPE and ICP-AES. AB - A method for the separation of tetrachloroplatinate PtCl(4)(2-), and hexachloroplatinate PtCl(6)(2-), by solid-phase extraction, using a Dowex 1x10 anion exchange resin is proposed. The sequential elution and separation of PtCl(4)(2-), and PtCl(6)(2-) is achieved using selective complexing agents. The eluates, containing Pt(II) and Pt(IV) were analyzed by inductively coupled plasma atomic emission spectroscopy (ICP-AES). Recoveries of 102% for PtCl(4)(2-) and 94% for PtCl(6)(2-) and detection limit of 15 ng g(-1) were achieved. Using this method determination of Pt(II) and Pt(IV) in soil samples, spiked with different platinum species was performed. The comparison with GFAAS determination showed a very good agreement. PMID- 18420042 TI - Improved identification of metabolites in complex mixtures using HSQC NMR spectroscopy. AB - The automated and robust identification of metabolites in a complex biological sample remains one of the greatest challenges in metabolomics. In our experiments, HSQC carbon-proton correlation NMR data with a model that takes intensity information into account improves upon the identification of metabolites that was achieved using COSY proton-proton correlation NMR data with the binary model of [Y. Xi, J.S. de Ropp, M.R. Viant, D.L. Woodruff, P. Yu, Metabolomics, 2 (2006) 221-233]. In addition, using intensity information results in easier-to-interpret "grey areas" for cases where it is not clear if the compound might be present. We report on highly successful experiments that identify compounds in chemically defined mixtures as well as in biological samples, and compare our two-dimensional HSQC analyses against quantification of metabolites in the corresponding one-dimensional proton NMR spectra. We show that our approach successfully employs a fully automated algorithm for identifying the presence or absence of predefined compounds (held within a library) in biological HSQC spectra, and in addition calculates upper bounds on the compound intensities. PMID- 18420043 TI - Modelling aroma of three Italian red wines by headspace-mass spectrometry and potential functions. AB - The aromas of 41 samples of wine from two Italian regions, Piedmont and Tuscany, were analysed by headspace-mass spectrometry. Samples were from three Italian wines (Barbera, Dolcetto and Chianti) produced in the same vintage, from different grape varieties and producing zones. The headspace generating conditions were optimised by full factorial experimental design then chemometric techniques were applied to verify the discriminating power of headspace-mass spectrometry among the three wine aromas. The modelling method based on potential function, applied on the first nine significant components of the 201 measured m/z, revealed best discrimination among the three wine aromas: cross-validated mean prediction rate of 96.7% and mean prediction rate of 83.3% on external test sets were obtained. PMID- 18420044 TI - Use of a modified, high-sensitivity, anodic stripping voltammetry method for determination of zinc speciation in the North Atlantic Ocean. AB - Zinc speciation is considered to be an important determinant of the biological availability of zinc. Yet in oceanic surface waters, characterization of zinc speciation is difficult due to the low concentrations of this essential micronutrient. In this study, an anodic stripping voltammetry method previously developed for the total determination of cadmium and lead was successfully adapted to the measurement of zinc speciation. The method differs from previous zinc speciation anodic stripping voltammetry methods in that a fresh mercury film is plated with each sample aliquot. The fresh film anodic stripping voltammetry method was compared to competitive ligand exchange cathodic stripping voltammetry in a profile from the North Atlantic Ocean. Results using the fresh film anodic stripping voltammetry method were similar to those determined using the cathodic stripping voltammetry method, though ligand concentrations determined by fresh film anodic stripping voltammetry were generally slightly higher than those determined by cathodic stripping voltammetry. There did not seem to be a systematic difference between methods for the estimates of conditional stability constants. The ligand concentration in the North Atlantic profile ranged from 0.9 to 1.5 nmol L(-1) as determined by fresh film anodic stripping voltammetry and 0.6 to 1.3 nmol L(-1) as determined by cathodic stripping voltammetry. The conditional stability constants determined by fresh film anodic stripping voltammetry were 10(9.8)-10(10.5) and by cathodic stripping voltammetry were 10(9.8)-10(11.3). PMID- 18420045 TI - Copper speciation by competing ligand exchange method using differential pulse anodic stripping voltammetry with ethylenediaminetetraacetic acid (EDTA) as competing ligand. AB - A technique has been developed to study chemical speciation of copper in freshwaters by competing ligand exchange (CLE) method using anodic stripping voltammetry (ASV) in the differential pulse (DP) mode with ethylenediaminetetraacetic acid (EDTA) as a competing ligand. The voltammetric behavior of Cu(II)-EDTA complex has been investigated using DPASV. When DPASV is used at an appropriate deposition potential, the inert Cu(II)-EDTA complex becomes electroactive, and is reduced directly. Furthermore, at the same deposition potential, Cu(II)-fuvic acid and Cu(II)-humic acid complexes do not contribute significantly to the analytical signal, which makes EDTA a suitable competing ligand in the determination of copper speciation using CLE-ASV. This method has been applied to freshwater samples from Rideau Canal (Ottawa, ON, Canada). The analysis of the copper titration data of these freshwater samples has indicated the presence of a very strong copper-binding ligand with a conditional stability constant of approximately 10(20) and a corresponding very high concentration (above 100 nM) of the ligand. PMID- 18420046 TI - Application of electrospray ionization tandem mass spectrometry for the rapid and sensitive determination of cobalt in urine. AB - Recently, cobalt (Co) is reported to be taken as a supplement by athletes for improving anaerobic performance. For the diagnosis of abuse, the limit of detection (LOD) of Co in the analysis should be lower than the concentrations of Co in plasma and urine of normal persons. A simple, rapid and sensitive method has been developed for the determination of Co in urine. Co was complexed with diethyldithiocarbamate (DDC) and extracted with isoamyl alcohol in the presence of citric acid. The detection of Co was achieved by injecting a 1-microL aliquot of isoamyl alcohol containing Co-DDC complex directly into an electrospray ionization tandem mass spectrometric (ESI-MS-MS) instrument without chromatographic separation. The quantification was performed using selected reaction monitoring at m/z 291 of the product ion Co(C(4)H(10)NCS)(2)(+) which was produced by collision-induced dissociation from the precursor ion Co(DDC)(2)(+) at m/z 355. ESI-MS-MS data were obtained in less than 10 min with an LOD of 0.05 microg L(-1) and a linear calibration range of 0.1-100 microg L( 1) using 10 microL of urine. The procedure was validated with certified reference materials (SRM 2670a and SRM 1643e). This method is suitable for the analysis of Co in the laboratories already equipped with an ESI-MS-MS instrument. PMID- 18420047 TI - Online stable carbon isotope ratio measurement in formic acid, acetic acid, methanol and ethanol in water by high performance liquid chromatography-isotope ratio mass spectrometry. AB - A suitable analysis condition was determined for high performance liquid chromatography-isotope ratio mass spectrometry (HPLC-IRMS) while making sequential measurements of stable carbon isotope ratios of delta(13)C in formic acid, acetic acid, methanol and ethanol dissolved in water. For this online column separation method, organic reagents are not applicable due to carbon contamination; thus, water and KH(2)PO(4) at low concentrations were tested as mobile phase in combination with a HyPURITY AQUASTAR column. Formic acid, acetic acid, methanol and ethanol were separated when 2 mM KH(2)PO(4) aqueous solution was used. Under the determined analysis condition for HPLC-IRMS, carbon concentrations could be measured quantitatively as well as carbon isotope ratio when carbon concentration was higher than 0.4 mM L for each chemical. PMID- 18420048 TI - Studies on the mechanism of the peroxyoxalate chemiluminescence reaction: part 2. Further identification of intermediates using 2D EXSY 13C nuclear magnetic resonance spectroscopy. AB - Further consideration has been given to the reaction pathway of a model peroxyoxalate chemiluminescence system. Again utilising doubly labelled oxalyl chloride and anhydrous hydrogen peroxide, 2D EXSY (13)C nuclear magnetic resonance (NMR) spectroscopy experiments allowed for the characterisation of unknown products and key intermediate species on the dark side of the peroxyoxalate chemiluminescence reaction. Exchange spectroscopy afforded elucidation of a scheme comprised of two distinct mechanistic pathways, one of which contributes to chemiluminescence. (13)C NMR experiments carried out at varied reagent molar ratios demonstrated that excess amounts of hydrogen peroxide favoured formation of 1,2-dioxetanedione: the intermediate that, upon thermolysis, has been long thought to interact with a fluorophore to produce light. PMID- 18420049 TI - Label-free cell-based assay using localized surface plasmon resonance biosensor. AB - For an understanding of the life activities, the analysis of cells, which are the smallest units of life form, has a significant impact on biology and biotechnology. In this study, we propose a novel label-free cell-based assay that is based on localized surface plasmon resonance (LSPR) biosensor, which is excited using core-shell structured nanoparticle layer substrate. To demonstrate the promising properties of our LSPR-based label-free cell-based assay, we performed the detection of the cell metabolites using the isolated cells from mouse thymus. For detection of the cellular metabolites, the refractive index change by the specific interaction between the antigen and antibody was detected on the antibody immobilized LSPR-based biosensor. Using our LSPR-based biosensor, the optical characteristics were monitored for the detection of specific reactions between antibody and cell metabolites. As a result, the detection limit of this antibody immobilized LSPR-based biosensor was 10 pg mL(-1). Furthermore, the time-course analysis of cell metabolisms using the isolated cells from mouse thymus was also achieved. From these results, the LSPR-based biosensor provides a promising platform with attractive advantages for the detection of biomolecular interactions at low-cost in a simplified experimental set-up with a low sample volume. PMID- 18420050 TI - Determination of aliphatic amines by high-performance liquid chromatography amperometric detection after derivatization with naphthalene-2,3 dicarboxaldehyde. AB - A simple and sensitive liquid chromatographic method has been developed for the determination of low molecular weight aliphatic amines after their pre-column derivatization with naphthalene-2,3-dicarboxaldehyde (NDA). Derivatization conditions, including the NDA concentration, reaction pH and reaction time have been investigated for method optimization. The chromatographic separation of five amines was performed on ABZ PLUS column using mobile phase of methanol-water (80:20, v/v) at a flow rate of 0.2 mL min(-1). The detection was carried out with a 6 mm glassy carbon electrode at the applied potential of 0.7 V versus Ag/AgCl reference electrode. The detection limits were between 23.3 and 34.4 nmol L(-1) of amines with a sample injection volume of 2 microL. The present method was applied for the determination of aliphatic amines in lake water. The recovery ranged 52.2-127.9%. The RSD in analytes retention time was less than 0.3% and 2.4% for intra- and inter-day analyses, respectively. The RSD in peak area was below 5.8% for both intra-day and inter-day analyses. The total analysis was completed within 20 min. PMID- 18420051 TI - Quantitative analysis of flavonoids and phenolic acids in Arnica montana L. by micellar electrokinetic capillary chromatography. AB - Arnica montana preparations have been used in Europe for centuries to treat skin disorders. Among the biologically active ingredients in the flower heads of the plant are sequiterpenes, flavonoids and phenolic acids. For the simultaneous determination of compounds belonging to the latter two groups a micellar electrokinetic capillary chromatography (MEKC) method was developed and validated. By using an electrolyte solution containing 50 mM borax, 25 mM sodium dodecyl sulfate and 30% of acetonitrile the separation of seven flavonoids and four caffeic acid derivatives was feasible in less than 20 min. The optimized system was validated for repeatability (sigma(rel) < or = 4.4%), precision (inter day sigma(rel) < or = 8.13%, intra-day sigma(rel) < or = 4.32%), accuracy (recovery rates from 96.8 to 102.4%), sensitivity (limit of detection (LOD) < or = 4.5 microg mL(-1)) and linearity (R(2) > or = 0.9996), and then successfully applied to assay several plant samples. In all of them the most dominant flavonoid was found to be quercetin 3-O-glucuronic acid, whereas 3,5 dicaffeoylquinic acid was the major phenolic acid; the total content of flavonoids and phenolic acids varied in the samples from 0.60 to 1.70%, and 1.03 to 2.24%, respectively. PMID- 18420052 TI - Determination of fluoxetine in plasma by gas chromatography-mass spectrometry using stir bar sorptive extraction. AB - This article presents a method employing stir bar sorptive extraction (SBSE) with in situ derivatization, in combination with either thermal or liquid desorption on-line coupled to gas chromatography-mass spectrometry for the analysis of fluoxetine in plasma samples. Ethyl chloroformate was employed as derivatizing agent producing symmetrical peaks. Parameters such as solvent polarity, time for analyte desorption, and extraction time, were evaluated. During the validation process, the developed method presented specificity, linearity (R(2)>0.99), precision (R.S.D.<15%), and limits of quantification (LOQ) of 30 and 1.37 pg mL( 1), when liquid and thermal desorption were employed, respectively. This simple and highly sensitive method showed to be adequate for the measurement of fluoxetine in typical and trace concentration levels. PMID- 18420053 TI - Molecular beacons: a real-time polymerase chain reaction assay for detecting Escherichia coli from fresh produce and water. AB - Molecular beacons (MBs) are oligonucleotide probes that fluoresce upon hybridization. The development of a real-time polymerase chain reaction (PCR) assay to detect the presence of Escherichia coli using these fluorogenic reporter molecules is reported. MBs were designed to recognize a 19-bp region of the uid A gene, coding for an enzyme beta-glucuronidase. The specificity of the MB-based PCR assay was evaluated for various E. coli strains as well as bacteria species that are present in nature. The capability of the assay to detect E. coli in drinking water and produce was demonstrated. Positive detection of E. coli was demonstrated when >10(1) CFU mL(-1) (colony forming unit) was present in the water samples and fresh produce after 18 h of enrichment. These assays could be carried out entirely in sealed PCR tubes, enabling rapid and semiautomated detection of E. coli in food and environmental samples. PMID- 18420054 TI - Amperometric sensors based on poly(3,4-ethylenedioxythiophene)-modified electrodes: discrimination of white wines. AB - The voltammetric responses on selected white wines of different vintages and origins have been systematically collected by three different modified electrodes, in order to check their effectiveness in performing blind analysis of similar matrices. The electrode modifiers consist of a conducting polymer, namely poly(3,4-ethylenedioxythiophene) (PEDOT) and of composite materials of Au and Pt nanoparticles embedded in a PEDOT layer. Wine samples have been tested, without any prior treatments, with differential pulse voltammetry technique. The subsequent chemometric analysis has been carried out both separately on the signals of each sensor, and on the signals of two or even three sensors as a unique set of data, in order to check the possible complementarity of the information brought by the different electrodes. After a preliminary inspection by principal component analysis, classification models have been built and validated by partial least squares-discriminant analysis. The discriminant capability has been evaluated in terms of sensitivity and specificity of classification; in all cases quite good results have been obtained. PMID- 18420055 TI - [Hallux valgus surgery in 2005. Conventional, mini-invasive or percutaneous surgery? Uni- or bilateral? Hospitalisation or one-day surgery?]. AB - There remains a good deal of controversy concerning forefoot surgery. Certain concepts such as conventional procedures, minimally invasive surgery, or percutaneous surgery are promoted because of their specific advantages including rapid recovery and compatibility with a short hospital stay or even outpatient surgery. Nevertheless, in 2005 many questions remain unanswered and highly variable practices have been basically founded on personal experience rather than scientific evidence. In addition, financial and lobbying pressure appears to have an influence on our choices, affecting the freedom of our therapeutic decision making. Developed over a long period, conventional surgery has proven reliability, reproducibility and adaptability. Procedures termed minimally invasive are defined by the limited incision. Percutaneous surgery is not less invasive than other procedures; the techniques are performed under indirect visual control and often assisted with more or less sophisticated radioscopic techniques depending on the surgeon's own experience. In our opinion, percutaneous surgery should be considered as a new concept based on rapid and functional results. Patients often raise the question of a bilateral procedure. For hallux valgus, there is no consensus on whether unilateral or bilateral procedures are better, the best solution depending on postoperative weight bearing and thus on the technique employed. From a cost expenditures point of view, bilateral procedures have an impact. For the advantages in terms of macroeconomy for professional incapacity, the question is less univocal for healthcare authorities. Advances in perioperative anesthesia and analgesia have enabled a broader approach to ambulatory surgery. Outpatient surgery appears to have benefits in terms of organization and economics. Variables studied were as follows: duration of hospital stay, postoperative edema, number of days of sick leave and preoperative and early and late postoperative pain. Patients who underwent minimally invasive procedures had a significantly shorter hospital stay compared with three other groups. For bilateral procedures, hospital stay on average was longer than in the two other groups. There was no correlation between postoperative edema and pain or between the degree of edema at 15 days and two months. Mean sick leave was 54.6 days. This was significantly shorter for percutaneous procedures compared with conventional surgery or minimally invasive techniques. Preoperative pain was noted four to five on the Visual Analogue Scale (VAS). There was no significant difference between the different groups as a function of the type of surgery performed. Statistically, there is very little difference in the short term between the different techniques. A much longer study would be necessary to obtain evidence to guide our practices. While there is certainly no reason to condemn one method or another, surgeons must be careful about the promises given to patients which are generally based on personal experience but not necessarily supported by rigorous scientific data. PMID- 18420056 TI - [Results of surgical treatment of congenital convex pes valgus (10 non-idiopathic feet)]. AB - PURPOSE OF THE STUDY: Congenital pes valgus is a rare and complex deformity of the foot raising serious diagnostic and therapeutic challenges. The purpose of our work was to present the surgical procedures used in our series and to analyze outcome. MATERIAL AND METHODS: Ten feet presenting congenital convex valgus treated surgically over a six-year period using the same operative technique were reviewed at minimum five years follow-up. Idiopathic deformities were excluded from this series. Deformities were secondary to arthrogryposis in five feet, a multiple malformative syndrome in four and diastematomyelia in one. The surgical technique used two approaches: a posteromedial incision to release the dorsal flexors, disinsert the tibialis posterior, open the talonavicular joint, release the Achilles tendon and release the posterior tibiotalar capsule; a lateral incision to lengthen the fibular tendons and perform an osteotomy of the anterior process of the calcaneum. A talonavicular pin and a calcaneocuboid pin maintained the correction. The tibialis posterior tendon was reinserted on the anterior aspect of the talonavicular capsule after incision of the dislocation chamber. RESULTS: Outcome was considered good in five cases and fair in five. Outcome was fair in the arthrogyrposis feet. Undercorrection was observed in two feet and valgus flatfoot in three. Talar necrosis occurred in one foot and navicular necrosis in two. DISCUSSION: Simultaneous correction of the different anomalies observed in the congenital convex foot was achieved in this series. The anatomic and functional results were satisfactory. We recommend avoiding overly extensive release in order to decrease the risk of talar and navicular necrosis. It is also important to check the reduction radiographically during the operation. Patients should use an orthesis several months postoperatively to avoid recurrence. PMID- 18420057 TI - [Results of open reduction and cortico-cancellous autograft of intra-articular calcaneal fractures according to Palmer]. AB - OBJECTIVE: To assess the mid-term patient outcome after operative treatment of a displaced intra-articular calcaneal fracture according to Palmer. MATERIALS AND METHODS: Eighteen operatively treated intra-articular fractures of the calcaneus in 16 patients (nine males, seven females) were retrospectively reviewed. Mean age at surgery was 35 years (range, 17-61 years). Two patients had a bilateral fracture. According to Rev Chir Orthop 79 (1993) 49-57, five fractures (28 percent) were classified as vertical, three as horizontal (17 percent), and 10 (55 percent) as combined. Mean preoperative Bohler angle was -4 degrees (range, 42-26 degrees ). An extended lateral approach to the calcaneus was used in each case. A cortical bone autograft was firmly impacted in the sub thalamic void after reduction. Stabilization was achieved by two or three axial Kirschner wires and one or two screw(s) inserted in a transverse position. Patients were evaluated at the last follow-up by physical examination and by the functional score described by Foot Ankle Int 15 (1994) 349-353. Bohler's angle was measured on the immediate postoperative radiographs and at the last follow-up, and compared when possible with the noninjured side. RESULTS: Follow-up ranged from 12 to 38 months (average, 23 months). One patient (one calcaneus) required a subtalar arthrodesis for advanced osteoarthritis. One patient (one calcaneus) experienced reflex sympathetic dystrophy. No skin complication was noted. At the last follow-up evaluation, mean Kitaoka score was 74 (range, 34-98). Functional score was considered good or excellent in 11 cases (64.7 percent), fair in three (17.6 percent) and poor in three (17.6 percent). Mean Bohler angle was 23.4 degrees (range, 5-40 degrees ) postoperatively and 22.7 degrees (range, 0-38 degrees ) at the last follow-up. A modest loss of correction was observed in four cases. Final Bohler angle was at least 85 percent comparatively to the healthy side in 10 cases among 14 unilateral fractures. Functional score correlated well with restoration of Bohler's angle. DISCUSSION AND CONCLUSION: Operative treatment of intra-articular calcaneal fractures according to Palmer provided encouraging results, since restoration of Bohler's angle was obtained. Main advantages include an absence of hardware prominence, resulting in an absence of skin complication, and a stable fixation. PMID- 18420058 TI - [Fractures of the lateral process of the talus. Retrospective study of 44 cases]. AB - PURPOSE OF THE STUDY: Fractures of the lateral process of the talus are often not diagnosed. The cohorts of the literature are small. The aim of the study was to analyse these fractures with a large group of patients. MATERIAL AND METHOD: Forty-four fractures in 43 patients were reviewed with an average follow-up of 17 months. The diagnosis had been made immediately in 14 cases and secondarily in 30 cases with a delay of 46 months. Patients had been evaluated with Kitaoka's score and radiographies using Hawkins classification. Fractures occurred during sport practise in 19 cases. The most frequent mechanism was association of dorsal flexion and pronation. There were associated lesions in 44% of cases. RESULTS: In the group of delayed diagnosis, we found 14 cases of associated pseudarthrodesis and sub-talar osteoarthritis, two cases of isolated pseudarthrodesis, two cases of isolated sub-talar osteoarthritis. After treatment in this group, the result at the last follow up was very good in 15 cases (50%), good in seven cases (23%), average in seven cases (23%) and bad in one case (4%). In the group of immediate diagnosis five of 14 patients had at least one complication: 29% of pseudarthrodesis and 29% of sub-talar osteoarthritis. After treatment in this group, the result at the last follow-up was very good in eight cases (58%), good in four cases (28%) and average in two cases (14%). Immediate diagnosis was correlated with better results at the last follow-up. In the sub-group of immediate diagnosis, among patients who had an orthopaedic treatment, the rate of secondarily surgery was 42%. CONCLUSION: The fracture of lateral process of the talus is quite frequent and occurs among young people. Spontaneous evolution is severe with two major complications: pseudarthrodesis and sub-talar osteo arthritis. Treatment is always required in case of displaced fracture. PMID- 18420059 TI - [Volar fixed angle plate LCP 3.5 for dorsally distal radius fracture. About 24 cases]. AB - PURPOSE OF THE STUDY: Fracture of the distal radius with posterior displacement is the most common wrist fracture. Pinning is generally proposed. The problem is that a secondary displacement may persist, even for the least complex fractures. With the advent of the fixed-angle plate with a self-locking screw, we considered using this method in combination with an anterior approach to the radius. We report our preliminary results. MATERIAL AND METHODS: From November 1 to March 31, 2003, among the 195 fractures of the distal radius with posterior displacement which we treated in our unit, 135 were treated with orthopedic reduction, 36 with Kapandji pinning and 24 with a T-shaped anterior locking compression plate (Synthes, LCP 3.5. During the early part of this series, the LCP was used for emergency situations in young adults with a posterior displacement exceeding 20 degrees . Starting in November 2002, the technique was also proposed for revisions for secondary displacements after orthopedic reduction or pinning procedures. Secondary displacement was noted on the standard X-rays and the clinical analysis included a measurement of muscle strength, the DASH score and the clinical evaluation score used by the SOFCOT symposium in November 2000. RESULTS: Twenty-four patients (14 female, 10 male, mean age: 54.5 years) presented 14 extra-articular fractures and ten articular fractures. At admission, mean frontal inclination of the radial glenoid was 13 degrees with a posterior displacement of 25.45 degrees and an ulnar variance measuring+4mm. Eighteen patients were reviewed at mean 16 months follow-up. Mean frontal inclination of the glenoid was 23.95 degrees with mean anteversion 5.45 degrees . Using the SOFCOT criteria, 11 outcomes were anatomic and seven presented moderate displaced healing. The wrist force (Jamar was 95% of the value measured on the healthy side. The mean DASH score was 38.47. Outcome was considered excellent for nine cases, good for six, fair for three and poor for zero. Complications were: reflex dystrophy (n=4), carpal tunnel syndrome (n=1), cheloid scar (n=1), glenoids in the pulse gutter (n=10), and secondary fracture along the upper border of the plate (n=3). CONCLUSION: This study demonstrated that, used on the anterior aspect of the radius, the locking compression plate provides satisfactory treatment for fractures of the distal radius with posterior displacement. Because of the angular stability, the reduction can be maintained over time so that secondary displacement is no longer a problem. This is a difficult technique which requires particular skill. We no longer recommend the LCP 3.5 plate due to the poorly adapted ergonometry, the late fracture over the plate and the fact that the material is not removed. PMID- 18420060 TI - [Intramedullary pinning of diaphyseal fractures of both forearm bones in adults: 46 cases]. AB - PURPOSE OF THE STUDY: The goal when treating shaft fractures of the forearm bones is to obtain good bone healing with good preservation of upper limb function. We report a retrospective (1995-2004) consecutive series of patients aged 32 years on average (range 16 to 92 years) who presented 46 shaft fractures of both forearm bones. MATERIAL AND METHODS: Using Hackethal's classification by sixths, the fracture involved the 3rd and 4th sixth in 39 cases (84.7%) for the radius and 33 (71.7%) for the ulna. Seventeen (37%) patients were trauma victims with multiple injuries. Fourteen fractures (30.4%) were open and all fractures were treated: 11 stage I (23.9%), two stage II (4.3%) and one stage III (2.3%). Intramedullary pinning was used in all cases for the radius and the ulna. In 27 patients (58.6%) pinning was achieved without open access to the focus. Pin diameter was 2.5mm for 38 cases (82.6%). Self-controlled rehabilitation exercises were started immediately without complementary immobilization. Outcome was assessed in 41 patients with a mean follow-up of 18 months (range three months to seven years). RESULTS: Functional outcome, according to Tscherne and Oestern, was very good or good in 31 cases (75.5%). For 35 cases (85.4%) bone healing was achieved at mean 3.5 months for the radius and four months for the ulna. Retarded healing was noted in four cases (9.7%) for the ulna. For the 41 patients reviewed, 11 (26.8%) presented axial misalignment of one of the forearm bones with an angle strictly greater than 10 degrees for three. There were six patients with nonunion (14.6%), of both bones in two and one in four. Radioulnar synostosis was observed in four cases (9.7%). No infections or recurrent fractures were noted. DISCUSSION: The localization of the fracture line affected the outcome. Among the four cases with a fracture in the 5th sixth of the radius, two developed misalignment with an angle strictly greater than 10 degrees and one presented a secondary displacement which required revision on day 15 for plate screw fixation of the radius. This might be due to the spreading corticals of the distal radius where pin stabilization would be less effective than for fractures in the 3rd or 4th sixths where the corticals run parallel. Outcome was less satisfactory when the fracture focus was opened. Four of the six nonunions and three of the four synostoses occurred among the 19 patients who had open pinning. This might be due to loss of the fracture hematoma. Analysis of the type of fixation has shown that our best results were obtained when we used a single 2.5 mm elastic pin. For the cases where we used a smaller pin (2 mm), we had three major misaligments, one nonunion involving both bones and one recurrent dislocation of the radial head. There were no infections or recurrent fractures in this series where the pin was removed in 27 patients (65.6%) at mean one year. CONCLUSION: Intramedullary pinning is an attractive alternative for the treatment of shaft fractures involving both forearm bones in adults. Best results are obtained if the fracture (situated outside the 5th sixth) can be stabilized without opening the focus with a single 2.5 mm pin. This method combines the advantages of closed osteosynthesis, that is, a simple nontraumatic procedure decreasing the risk of suppuration, and early return of function, limiting postoperative immobilization. It enables early sturdy bone healing with a low risk of recurrent fracture. PMID- 18420061 TI - [Acute hematogenous osteomyelitis of the obturator rim in seven children]. AB - PURPOSE OF THE STUDY: Acute osteomyelitis of the rim of the obturator foramen is rarely reported. The clinical presentation is atypical, making diagnosis a difficult task. We report a series of seven cases of osteomyelitis of the obturator rim which illustrate the different features of this localization. MATERIAL AND METHODS: The series included seven boys with osteomyelitis involving the obturator foramen. We reviewed the clinical history, the diagnostic approach and management. Complications and anatomic outcome were noted at mean three years follow-up (range one to 10 years). RESULTS: Mean age was 9.5 years (five to 12 years). Bilateral involvement was noted in one boy. Mean time from symptom onset to consultation was five days and mean time from consultation to hospitalization was eight days. No specific clinical presentation could be identified. Fever was not a constant feature. Mean body temperature was 38.6 degrees C and was not greater than 38.5 degrees C in four children. Symptoms were limited to hip pain in five cases and abdominopelvic pain was noted in two. Physical examination failed to trigger exquisite ischial or pubic pain in two patients. The osteomyelitis involved the ischiopubic ramus in four cases, the ischium in two with one bilateral case, and the pubis in one. Certain diagnosis was established as follows: MRI findings plus isolation of the pathogenic agent (n=4) ; the plain x-ray showed a defect in the ischiopubic ramus, MRI showed signs favoring osteomyelitis of the ischiopubic ramus and surgery evacuated a purulent collection (n=1) ; strong uptake of the obturator rim on scintigraphy (n=2). Medical treatment was given in all cases and surgery was performed in six patients. Outcome was noted at three years follow-up, range one to 10 years. All patients recovered normal function. There were no complications. DISCUSSION: Hematogenous osteomyelitis of the pelvis is exceptional (2-11 % of cases of osteomyelitis). Localization in the obturator rim is rare. We discuss the specific diagnostic and therapeutic features of this localization. Despite the controversy concerning the treatment of pelvic osteomyelitis and in particular cases involving the obturator rim, prognosis is generally good. No complications have been described in the literature specifically associated with osteomyelitis of the obturator rim. PMID- 18420062 TI - [Mirror hand deformity: a new phenotype with literature review]. AB - We report the case of a child who presented polydactyly with eight triphalangeal fingers, no thumb or radius and ulnar dimelia. Hand, wrist, forearm and elbow function was compromised, particularly due to wrist stiffness in flexion, the absence of forearm pronation supination and severe limitation of elbow motion. In addition, the child underwent surgery for pyloric hypertrophy and also presented a multicystic kidney. We present the clinical, anatomic, electromyographic, genetic and therapeutic aspects of this rare deformity and discuss data presented in the literature. PMID- 18420063 TI - [Acute carpal tunnel syndrome secondary to prolonged pressure on the wrist during enolic coma]. AB - We report a case of acute carpal tunnel syndrome caused by prolonged compression. A 40-year-old man was admitted for an acute carpal tunnel syndrome secondary to direct compression of the wrist which was blocked in supination under his thorax for ten hours during a period of alcoholic coma. Total sensorial anesthesia of the median nerve territory was noted. The emergency procedure consisted in simple opening of the carpal tunnel without nerve exploration due to the risk of bacterial contamination resulting from skin lesions, devascularization and postoperative fibrosis. Initially, the skin on the volar aspect of the wrist had the aspect of a second degree burn. The patient recovered nerve function the next day and the skin wound healed within 15 days. The patient was seen at consultation at 13 months and exhibited complete recovery of wrist and hand motion with normal thumb opposition and no signs of sensorial or motor deficit. The retinaculum of the flexor system must be opened to guarantee full nervous recovery. PMID- 18420064 TI - [Bulge of the scapholunate ligament: an arthro-CT sign of traumatic scapholunate instability]. AB - Tears of the scapholunate ligament are usually diagnosed during an arthro-CT or arthro-MRI examination. Typically, the contrast agent passes from one joint to the other. In certain cases, fibrous scar tissue devoid of any mechanical effect may block the passage and lead to an erroneous interpretation of the images. We reviewed retrospectively patients who underwent arthroscopic treatment for scapholunate tears and who had normal arthro-CT images. We searched for specific aspects suggestive of ligament injury. Analysis of the arthro-CT images demonstrated that a bulge of the scapholunate ligament could be noted in patients whose diagnosis of scapholunate ligament injury was established arthroscopically. We propose that appropriate lecture of wrist images can identify scapholunate ligament injury in patients wrongly considered to be free of wrist injury, but who actually have an occult injury to the scapholunate ligament. PMID- 18420065 TI - [Desmoplastic fibroblastoma of the foot]. AB - Desmoplastic fibroblastoma is a benign, rare, slow-growing soft tissue tumor which is found in a wide anatomic distribution, predominantly in adult males. The characteristic gross aspect is that of a typical cartilage-like tumor which histologically presents regular fibroblastic proliferation, often in a stellar shape within a dense collagen or myxo-collagen stroma. We report a new case observed in the foot and study the anatomic and clinical aspects of this rare entity. PMID- 18420066 TI - [A secondary bleeding false aneurysm of the superomedial genicular artery after distal femoral epiphysiodesis: a case report]. AB - A 16-year-old male developed a false aneurysm of the superomedial genicular artery five weeks after medial distal femoral epiphysiodesis. The aneurysm was revealed by inexhaustible bleeding from the medial aspect of the knee and the presence of acute hemorrhage which resolved spontaneously. Physical examination disclosed disunion of the surgical wound at the apex of a pulsatory tumefaction over the medial aspect of the knee. AngioCT enabled the diagnosis of a false aneurysm of the superomedial genicular artery. Emergency surgery was undertaken to drain the hematoma and exclude the false aneurysm by ligating the superomedial genicular artery. PMID- 18420067 TI - [Comments on: "low cost accuracy of the anterior pelvic plane to guide the position of the cup with imageless computer assistance: variation of position in 106 patients" (no. 5-2007)]. PMID- 18420069 TI - [Journal of Bone and Joint Surgery (Br), Lecture by James Scott M.D. - November 6, 2007]. PMID- 18420070 TI - [Signs, symptoms or syndromes?]. PMID- 18420071 TI - [Severity scores in dermatology]. PMID- 18420072 TI - [Community-acquired methicillin-resistant Staphylococcus aureus in a dermatology outpatient clinic]. AB - BACKGROUND: Community-acquired cutaneous infections caused by methicillin resistant Staphylococcus aureus (MRSA) are a growing concern. These bacteria may produce Panton-Valentine leucocidin potentially leading to necrotizing pneumonia. We studied the prevalence of MRSA and Panton-Valentine leucocidin in dermatology clinic outpatients in order to adapt therapy where possible. PATIENTS AND METHODS: This was a prospective study including all patients seen at a dermatology outpatient clinic between 1st March 2005 and 31st December 2006 and presenting mucocutaneous bacteriological samples. The main MRSA risk factors studied were frequent hospital consultations, hospitalization, antibiotic therapy within the last three months and community life. The following risk factors were also analysed, although less routinely: substance abuse, immunosuppression, diabetes mellitus, recent travel abroad and a history of similar lesions. RESULTS: One hundred and twenty-two patients were included in the study and 235 samples (143 lesion samples and 92 nasal swabs) were carried out and S. aureus was isolated in 68 patients (56%). Twelve patients had MRSA (17.6%); seven of these were normal outpatients but five attended frequent hospital consultations (7.3%). MRSA resistance rates were as follows: 64% to ofloxacin, 36% to amikacin and erythromycin, 27% to fusidic acid, 9.1% to sulfamethoxazole-trimethoprim and 0% to pristinamycin. Community life was the only significant risk factor for MRSA in this study (p=0.045). Four of the 11 MRSA strains tested produced Panton Valentine leucocidin. CONCLUSION: Dermatologists are increasingly faced with cutaneous infections caused by community-acquired MRSA. Bacterial samples should be taken routinely and probabilistic antibiotic therapy for MRSA instituted in severe infections. PMID- 18420073 TI - [Mucosal involvement in lymphomatoid papulosis: four cases]. AB - BACKGROUND: Lymphomatoid papulosis is a form of CD30+ cutaneous lymphoproliferation characterized by a benign chronic papulonodular eruption that regresses spontaneously. The clinical features contrast with the malignant histological aspect of lesions. Mucosal lesions are rare, with less than 10 published cases. We report four new cases and we highlight characteristic features of lesions at this particular site. PATIENTS AND METHODS: We report four cases of mucous lymphomatoid papulosis in three women aged 37, 38, and 71 years and one 66-year-old man. These cases were collated from three different hospitals: Orleans, Rouen and Caen. Mucosal lesions occurred after cutaneous eruption in two cases but remained isolated or preceded cutaneous lesions in the other two cases. The main site was the mouth in all four cases but one case also involved genital lesions. Two cases involved type A pathological features and two had type C features. Association with lymphoma was excluded on clinical, laboratory and radiological examination. One patient was treated with methotrexate (>7.5mg/week) and did not relapse. Of the three other untreated patients only one did not relapse (short 14-month follow-up). DISCUSSION: Recurrent oral ulcerations may be mucosal manifestations of lymphomatoid papulosis. This site does not appear to have any bearing on prognosis. PMID- 18420074 TI - [Cutaneous fissures in collodion babies: incidence and treatment]. AB - BACKGROUND: Collodion baby syndrome (CBS) is a pathological cutaneous condition present at birth and is due to the presence of a thick horny layer of the skin. Exfoliation begins early with drying and cracking of the collodion membrane. The cracks may either remain superficial or they may be deeper and affect the superficial dermis, in which case, fissures form. This study of CBS provides information on the clinical aspect of fissures, their incidence, their pathological consequences and therapeutic approaches. PATIENTS AND METHODS: In our study, diagnosis of CBS was made clinically based on the presence of neonatal collodion membrane. Identification of typical cracks and fissures was made on clinical examination and their site, chronology and consequences were assessed. Routine bacteriological examination of fissures was performed twice weekly on a clinically infected specimen taken from an inguinal fissure. The therapeutic protocol for CBS has been validated and involved use of sterile vaseline oil. Fissures were disinfected. RESULTS: Cracks are a constant feature. Fissures were seen in 20 of the 33 cases of CBS with the site of predilection being large skinfolds. Morphine was necessary for pain relief in three cases. Pathogenic organisms were isolated in all cases of inflammatory fissures. Dissemination of septicaemia was confirmed in four cases and the offending organism was isolated from the inflammatory fissures in all cases. Candida albicans was present in all cases of fissures in the inguinal folds or between the buttocks. Keratotic adhesions occurred after healing of digital fissures and a surgical procedure was required in this event. DISCUSSION: Fissures are lesions occurring secondarily to cracks and they were seen in 20 of the 33 cases of CBS. These secondary lesions are rarely mentioned in the literature and the reasons for this oversight are discussed. Fissures are potential complications in all states of CBS, particularly where the collodion is thick. Topical treatment does not prevent transformation of cracks to fissures in all cases. Where fissures are not inflammatory, routine prescription of oral antibiotics is not always necessary. Regular bacteriological monitoring at several different fissure sites allows selection of appropriate antibiotic therapy. The main therapeutic goal in CBS is to treat painful fissures and superinfection. PMID- 18420075 TI - [Sezary syndrome and Leser-Trelat sign: a chance association?]. AB - BACKGROUND: Leser-Trelat sign involves the combined sudden onset of seborrheic keratosis and cancer. However, some doubt surrounds the existence of this syndrome. We report a case of Leser-Trelat sign that led to the discovery of Sezary syndrome. PATIENTS AND METHODS: A 59-year-old woman presented generalized pruritus with secondary appearance of multiple seborrheic keratosis. Leser-Trelat sign was diagnosed and 20 months later, Sezary syndrome was discovered. Extracorporeal photopheresis was initiated, after which there was a marked reduction in the patient's pruritus, erythroderma and numbers of seborrheic keratoses and Sezary cells. DISCUSSION: Leser-Trelat sign is often associated with gastric carcinoma or lymphoproliferative tumours. Rampen and Schwengle [J Am Acad Dermatol 21 (1989) 50-5] have thrown doubt on this entity because of the "subjective" definition, the frequent dissociation between the course of the tumour and that of the seborrheic keratosis, the disparity between the frequency with which rapid onset seborrheic keratosis is seen and the rarity of cases in which this phenomenon reveals a tumour and the absence of association with any specific type of malignancy. The time between diagnosis of Sezary syndrome and cutaneous symptoms of Leser-Trelat sign appears very long in the present case. In the absence of any established physiopathology, it is impossible to prove any direct link between these two syndromes. Leser-Trelat sign remains controversial. Knowledge of its pathogenesis could help determine whether Leser-Trelat sign should or should not be considered a paraneoplastic syndrome. PMID- 18420076 TI - [Sweet's syndrome and phenylbutazone-induced sialadenitis]. AB - BACKGROUND: Phenylbutazone frequently induces a range of potentially dangerous adverse reactions. We report a case of Sweet's syndrome with sialadenitis induced by phenylbutazone. CASE-REPORT: A 54-year-old woman presented lumbar pains treated with phenylbutazone for three days. Six days later, she exhibited inflammation of the submaxillary and parotid salivary glands, followed by an erythematous, oedematous, pustular and febrile eruption, with failure of antibiotic therapy. Laboratory data showed leukocytosis and neutrophilia, anaemia, an elevated platelet count and liver dysfunction. The infectious and autoimmune tests were negative. The skin biopsy confirmed Sweet's syndrome. Clinical and biological abnormalities resolved on administration of systemic steroids. DISCUSSION: Phenylbutazone-induced sialadenitis is rare and presents unrecognized adverse effects that may be associated with a systemic reaction. In the present case report, Sweet's syndrome met the criteria for drug-induced Sweet's syndrome. There appears to have been a systemic reaction caused by a hypersensitivity mechanism, in the same way as sialadenitis. PMID- 18420077 TI - [Fluindione-induced acute generalised exanthematous pustulosis confirmed by patch testing]. AB - BACKGROUND: Fluindione is an oral anticoagulant belonging to the vitamin K antagonist class. Although fluindione is very widely prescribed in France, few cases of cutaneous drug reactions have been reported. Below we describe a case of acute generalised exanthematous pustulosis due to fluindione as confirmed by patch-testing. PATIENTS AND METHODS: A 70-year-old woman was hospitalised for diffuse erythematous and pustular rash 48hours after initiation of fluindione treatment for cardiac arrhythmia. A diagnosis of fluindione-induced acute generalised exanthematous pustulosis was made. After withdrawal of fluindione, the eruption cleared up within eight days. Warfarin was then used without skin reaction. Subsequent patch-tests were positive for fluindione. DISCUSSION: These signs were consistent with fluindione-induced acute generalised exanthematous pustulosis. A causative role of fluindione is very likely in view of the rapid onset after initiation, improvement after withdrawal and positive patch tests. Skin patch-testing, which is easily performed, can be extremely helpful in determining a causal relationship with medication. PMID- 18420078 TI - [Intravascular B-cell lymphoma with febrile inflammatory lymphoedema of the lower limbs and lower back]. AB - BACKGROUND: Intravascular lymphomas are diffuse large-cell lymphomas belonging to a group of high-grade non-Hodgkin's lymphomas and are generally of phenotype B. They are rare and carry a severe prognosis. Clinical polymorphism is dominated by neurological and cutaneous involvement. PATIENTS AND METHODS: We report the case of an 80-year-old woman with cutaneous intravascular B-cell lymphoma as revealed by an isolated episode of febrile bilateral inflammatory lymphoedema. Following combined chemotherapy with rituximab and mini-CHOP (cyclophosphamide, adriamycin, oncovin and prednisone), complete remission was obtained rapidly, with no relapse at two years. DISCUSSION: Diagnosis of these tumours is rendered difficult by the clinical polymorphism and multifocal nature of lymphocytic proliferations. In the present case, diagnosis was based on histology results since presentation of the disease in the form of bilateral inflammatory oedema of the lower limbs is not sufficient to establish lymphoma. Combined rituximab and polychemotherapy comprising a CHOP regimen appears to yield the best results. PMID- 18420079 TI - [Leukaemia cutis during acute aleukaemic myeloid leukaemia in a two-month-old infant]. AB - BACKGROUND: Congenital cutaneous leukaemia is rare. PATIENTS AND METHODS: A two month-old girl presented bluish cutaneous macules of the trunk, histological examination of which suggested acute myeloid leukaemia (LAM B 5). The blood picture was negative for circulating tumour cells and the outcome under chemotherapy was favourable at one year of follow-up. DISCUSSION: The prognosis of congenital leukaemia is serious. Aleukaemic congenital leukaemia is seen occasionally but is rare. The existence of multiple cutaneous tumours in newborn infants raises the possibility of TORCH infection and of other malignant tumours such as nephroblastoma or neuroblastoma. PMID- 18420080 TI - [Extensive oral condylomas treated by in situ cidofovir injection in an HIV patient]. AB - BACKGROUND: Human papillomavirus infections are difficult to treat and have a high rate of recurrence, especially in a setting of human immunodeficiency (HIV) infection. Moreover, there is no standard treatment for oral condylomas. PATIENTS AND METHODS: We report the partial success of in situ injections of cidofovir in an HIV patient, presenting extensive oral condylomas. The injections were well tolerated and the response was still present at one year while the immune status of the patient was unchanged. DISCUSSION: The efficacy of topical cidofovir against condyloma acuminata has been reported and the value of in situ cidofovir injections for the treatment of laryngeal papillomatosis is well established. This case report shows the need for further investigation of in situ cidofovir injections as an alternative treatment for human papillomavirus lesions that are difficult to treat because of both site and extension. PMID- 18420081 TI - [Linear IgA bullous dermatosis associated with spondylarthropathy in a child]. PMID- 18420082 TI - [Amifostine-induced Stevens-Johnson syndrome during radiotherapy]. PMID- 18420083 TI - [Glucantime reaction with fatal outcome following treatment of cutaneous leishmaniasis]. PMID- 18420084 TI - [Cutaneous manifestations of chronic renal failure]. PMID- 18420085 TI - [Bevacizumab therapy for Poems syndrome]. PMID- 18420086 TI - [Umbilicated papules of the head, neck and trunk in an HIV-positive patient]. PMID- 18420087 TI - [Epidemiology, clinical picture and treatment of molluscum contagiosum: literature review]. PMID- 18420088 TI - [Pigmented Bowen's disease]. PMID- 18420089 TI - [Administration procedure for meopa gas: Entonox, Kalinox or Medimixe]. PMID- 18420090 TI - [Focal dermal hypoplasia: PORCN deficiency?]. PMID- 18420091 TI - [Pseudocondylomatous lymphangiectasia of the vulva secondary to lipoedema]. PMID- 18420092 TI - [Asymptomatic subpreputial nodule]. PMID- 18420093 TI - [Escitalopram, labial pigmentation and lichenoid eruption]. PMID- 18420094 TI - Pulmonary arterial hypertension. AB - Significant advances in the treatment of pulmonary arterial hypertension (PAH) have occurred over the last 10 years, starting with the approval of epoprostenol in 1998. Subsequently, multiple additional medications have received approval, including a subcutaneous prostacyclin, an inhaled prostacyclin, and oral medications in 2 separate classes. Over this same period, the classification of pulmonary hypertension has been revised with changes including the substitution of the term idiopathic for primary PAH and an expanded list of conditions felt to be associated with the development of PAH. Long-term follow-up studies have provided better information on prognosis and expected outcomes with treatment, with particularly valuable data on reassessment of prognosis after treatment with epoprostenol. Combination therapy is more frequently being used, and limited data on novel therapies such as stem cell transplantation have been published. The purpose of this review is to describe the current state of evidence for the diagnosis, prognosis, and treatment of the patient with PAH. PMID- 18420095 TI - In search of the "vulnerable plaque": can it be localized and will focal regional therapy ever be an option for cardiac prevention? AB - The search to find the location of future plaque ruptures or plaque erosions leading to myocardial infarction (so-called "vulnerable plaques") is an important area of cardiovascular research. Systemic therapy, including use of statins, targets the vulnerable patient. However, adverse events cannot be completely eliminated with the appropriate application of systemic therapies and thus has given rise to the possibility of local or regional therapy of "vulnerable plaques" to prevent future events. Until now, no criteria have been developed for consideration of this therapy. For such a strategy to work, there should be several prerequisites. These involve the identification of susceptible lesions, the number of lesions, their natural history, and proof that an interventional technique is preferable to medical therapy alone. The greatest deficiency relates to the fact that until the natural history of presumed "vulnerable plaques" is known one can never truly identify what constitutes a "vulnerable plaque." Much work needs to be done in this area, but ongoing and new trials should provide important information that could potentially change drastically how coronary artery disease is diagnosed and treated. PMID- 18420096 TI - A novel bioresorbable polymer paclitaxel-eluting stent for the treatment of single and multivessel coronary disease: primary results of the COSTAR (Cobalt Chromium Stent With Antiproliferative for Restenosis) II study. AB - OBJECTIVES: The aim was to compare safety and effectiveness of the CoStar drug eluting stent (DES) (Conor MedSystems, Menlo Park, California) with those of the Taxus DES (Boston Scientific, Maple Grove, Minnesota) in de novo single- and multivessel percutaneous coronary intervention (PCI). BACKGROUND: Paclitaxel elution from a stent coated with biostable polymer (Taxus) reduces restenosis after PCI. The CoStar DES is a novel stent with laser-cut reservoirs containing bioresorbable polymer loaded to elute 10 microg paclitaxel/30 days. METHODS: Patients undergoing PCI for a single target lesion per vessel in up to 3 native epicardial vessels were randomly assigned 3:2 to CoStar or Taxus. Primary end point was 8-month major adverse cardiac events (MACE), defined as adjudicated death, myocardial infarction (MI), or clinically driven target vessel revascularization (TVR). Protocol-specified 9-month angiographic follow-up included 457 vessels in 286 patients. RESULTS: Of the 1,700 patients enrolled, 1,675 (98.5%) were evaluable (CoStar = 989; Taxus = 686), including 1,330 (79%) single-vessel and 345 (21%) multivessel PCI. The MACE rate at 8 months was 11.0% for CoStar versus 6.9% for Taxus (p < 0.005), including adjudicated death (0.5% vs. 0.7%, respectively), MI (3.4% vs. 2.4%, respectively), and TVR (8.1% vs. 4.3%, respectively). Per-vessel 9-month in-segment late loss was 0.49 mm with CoStar and 0.18 mm with Taxus (p < 0.0001). Findings were consistent across pre specified subgroups. CONCLUSIONS: The CoStar DES is not noninferior to the Taxus DES based on per-patient clinical and per-vessel angiographic analyses. The relative benefit of Taxus is primarily attributable to reduction in TVR. Follow up to 9 months showed no apparent difference in death, MI, or stent thrombosis rates. PMID- 18420098 TI - Vascular effects of diets, especially plant sterol ester consumption. PMID- 18420097 TI - Vascular effects of diet supplementation with plant sterols. AB - OBJECTIVES: The purpose of this study was to evaluate vascular effects of diet supplementation with plant sterol esters (PSE). BACKGROUND: Plant sterol esters are used as food supplements to reduce cholesterol levels. Their effects on endothelial function, stroke, or atherogenesis are not known. METHODS: In mice, plasma sterol concentrations were correlated with endothelial function, cerebral lesion size, and atherosclerosis. Plasma and tissue sterol concentrations were measured by gas-liquid chromatography-mass spectrometry in 82 consecutive patients with aortic stenosis. RESULTS: Compared with those fed with normal chow (NC), wild-type mice fed with NC supplemented with 2% PSE showed increased plant sterol but equal cholesterol plasma concentrations. The PSE supplementation impaired endothelium-dependent vasorelaxation and increased cerebral lesion size after middle cerebral artery occlusion. To test the effects of cholesterol lowering by PSE, apolipoprotein E (ApoE)-/- mice were randomized to Western-type diet (WTD) with the addition of PSE or ezetimibe (EZE). Compared with WTD, both interventions reduced plaque sizes; however, WTD + PSE showed larger plaques compared with WTD + EZE (20.4 +/- 2.1% vs. 10.0 +/- 1.5%). Plant sterol plasma concentration strongly correlated with increased atherosclerotic lesion formation (r = 0.50). Furthermore, we examined plasma and aortic valve concentrations of plant sterol in 82 consecutive patients with aortic stenosis. Patients eating PSE supplemented margarine (n = 10) showed increased plasma concentrations and 5-fold higher sterol concentrations in aortic valve tissue. CONCLUSIONS: Food supplementation with PSE impairs endothelial function, aggravates ischemic brain injury, effects atherogenesis in mice, and leads to increased tissue sterol concentrations in humans. Therefore, prospective studies are warranted that evaluate not only effects on cholesterol reduction, but also on clinical endpoints. PMID- 18420099 TI - Lipid-altering efficacy and safety of ezetimibe/simvastatin coadministered with extended-release niacin in patients with type IIa or type IIb hyperlipidemia. AB - OBJECTIVES: This study evaluated the safety and lipid-altering efficacy of ezetimibe/simvastatin (E/S) coadministered with extended-release niacin (N) in patients with type IIa or IIb hyperlipidemia. BACKGROUND: Current guidelines recommend consideration of combination drug therapy to achieve optimal low density lipoprotein cholesterol (LDL-C) lowering and broader lipid-altering effects when treating hypercholesterolemic patients at high risk for atherosclerotic cardiovascular events. METHODS: In this 24-week multicenter, randomized, double-blind study, 1,220 type IIa or IIb hyperlipidemic patients were randomized to treatment with E/S (10/20 mg/day) + N (titrated to 2 g/day), or N (titrated to 2 g/day), or E/S (10/20 mg/day). Changes from baseline in LDL-C (primary) and other secondary variables were assessed in the completers and modified intent-to-treat populations. RESULTS: Coadministered E/S with N resulted in significantly greater reductions in LDL-C, non-high-density lipoprotein cholesterol, triglycerides, apolipoprotein B, and lipid/lipoprotein ratios, compared with either agent alone (p < 0.001). The combination increased levels of apolipoprotein A-I and high-density lipoprotein cholesterol significantly more than E/S (p < 0.001), and reduced high-sensitivity C-reactive protein levels significantly more than N (p = 0.005). A significantly greater percentage of patients discontinued the study in the N (25.0%) and N + E/S (23.3%) groups, compared with E/S (9.6%, p < 0.001) because of clinical adverse experiences (primarily flushing). Incidences of other clinical and laboratory adverse experiences (liver-, muscle-, and gastrointestinal-related) were similar for all groups. CONCLUSIONS: Combination treatment with E/S plus N showed superior lipid altering efficacy compared with N or E/S in type IIa or IIb hyperlipidemia patients and was generally well tolerated aside from N-associated flushing. This combination offers an effective, broad, lipid-altering therapy with improvements in lipid effects beyond LDL-C in these patients. (To Evaluate Ezetimibe/Simvastatin and Niacin [Extended Release Tablet] in Patients With High Cholesterol. PMID- 18420100 TI - Diabetic retinopathy and risk of heart failure. AB - OBJECTIVES: The purpose of this study was to examine the association of diabetic retinopathy with incident heart failure (HF). BACKGROUND: Microvascular disease might play a more prominent role in the pathogenesis of diabetic cardiomyopathy, a major cause of HF in diabetes. Whether diabetic retinopathy, a microvascular complication of diabetes, predicts HF is unclear. METHODS: A population-based study included 1,021 middle-aged type 2 diabetic persons with normal renal function and free of clinical coronary heart disease or HF at baseline. Diabetic retinopathy signs were graded from retinal photographs. Incident HF events were prospectively identified from hospital stay and death records. RESULTS: There were 125 (12.8%) participants with diabetic retinopathy. After 9-year follow-up, 106 (10.1%) participants developed incident HF events. Persons with retinopathy were more likely to develop HF (cumulative incidence of 21.6%) than those without retinopathy (cumulative incidence of 8.5%). After controlling for age, gender, race, smoking, diabetes duration, insulin use, blood pressure, lipid profile, and other risk factors, participants with retinopathy had more than 2.5-fold higher risk of developing HF than those without retinopathy (hazard ratio [HR] 2.71; 95% confidence interval [CI] 1.46 to 5.05). This association remained significant after further adjustments for glycemic control, carotid atherosclerosis, and serum markers of endothelial dysfunction (HR 2.20, 95% CI 1.08 to 4.47). CONCLUSIONS: The presence of diabetic retinopathy signifies an excess risk of HF, independent of known risk factors. This further supports a contribution of microvascular disease to the development of HF in people with diabetes. PMID- 18420101 TI - The eye as an indicator of heart failure in diabetic patients. PMID- 18420102 TI - The salvaged area at risk in reperfused acute myocardial infarction as visualized by cardiovascular magnetic resonance. AB - OBJECTIVES: We aimed to characterize the tissue changes within the perfusion bed of infarct-related vessels in patients with acutely reperfused myocardial infarction (MI) using cardiovascular magnetic resonance (CMR). BACKGROUND: Even in successful early revascularization, intermittent coronary artery occlusion affects the entire perfusion bed, also referred to as the area at risk. The extent of the salvaged area at risk contains prognostic information and may serve as a therapeutic target. Cardiovascular magnetic resonance can visualize the area at risk; yet, clinical data have been lacking. METHODS: We studied 92 patients with acute MI and successful reperfusion 3 +/- 3 days after the event and 18 healthy control subjects. Breath-hold T2-weighted and contrast-enhanced ("late enhancement") CMR were used to visualize the reversible and the irreversible myocardial injury, respectively. RESULTS: All reperfused infarcts consistently revealed a pattern with both reversibly and irreversibly injured tissue. In contrast to the infarcted area, reversible damage was always transmural, exceeding the infarct in its maximal extent by 16 +/- 11% (absolute difference of the area of maximal infarct expansion 38 +/- 15% vs. 22 +/- 10%; p < 0.0001). None of the controls had significant T2 signal intensity abnormalities. CONCLUSIONS: In patients with reperfused MI, CMR visualizes both reversible and irreversible injury. This allows for quantifying the extent of the salvaged area after revascularization as an important parameter for clinical decision-making and research. PMID- 18420103 TI - Long-term prognosis of patients with peripheral arterial disease: a comparison in patients with coronary artery disease. AB - OBJECTIVES: This study was designed to compare the long-term outcomes of patients with peripheral arterial disease (PAD) with a risk factor matched population of coronary artery disease (CAD) patients, but without PAD. BACKGROUND: The PAD is considered to be a risk factor for adverse late outcome. METHODS: A total of 2,730 PAD patients undergoing vascular surgery were categorized into groups: 1) carotid endarterectomy (n = 560); 2) elective abdominal aortic surgery (AAA) (n = 923); 3) acute AAA surgery (r-AAA) (n = 200), and 4) lower limb reconstruction procedures (n = 1,047). All patients were matched using the propensity score, with 2,730 CAD patients who underwent coronary angioplasty. Survival status of all patients was obtained. In addition, the cause of death and complications after surgery in PAD patients were noted. The Kaplan-Meier method was used to compare survival between the matched PAD and CAD population and the different operation groups. Prognostic risk factors and perioperative complications were identified with the Cox proportional hazards regression model. RESULTS: The PAD patients had a worse long-term prognosis (hazard ratio 2.40, 95% confidence interval 2.18 to 2.65) and received less medication (beta-blockers, statins, angiotensin-converting enzyme inhibitors, aspirin, nitrates, and calcium antagonists) than CAD patients did (p < 0.001). Cerebro-cardiovascular complications were the major cause of long-term death (46%). Importantly, no significant difference in long-term survival was observed between the AAA and lower limb reconstruction groups (log rank p = 0.70). After vascular surgery, perioperative cardiac complications were associated with long-term cardiac death, and noncardiac complications were associated with all-cause death. CONCLUSIONS: Long-term prognosis of vascular surgery patients is significantly worse than for patients with CAD. The vascular surgery patients receive less cardiac medication than CAD patients do, and cerebro-cardiovascular events are the major cause of late death. PMID- 18420104 TI - Patch closure of muscular ventricular septal defects with a new hybrid therapy in a pig model. AB - OBJECTIVES: We evaluated a novel technique for hybrid patch closure of muscular ventricular septal defects (mVSDs) without cardiopulmonary bypass (CPB) in a pig model. BACKGROUND: So far, surgical and interventional therapies for mVSDs have been associated with significant morbidity, especially in newborns and infants. Thus, it is essential to develop new techniques. Hybrid therapy is an innovative approach for mVSDs that combines the advantages of surgical and interventional techniques. METHODS: Six pigs underwent left anterolateral thoracotomy to expose the left ventricle (LV). The mVSDs were created under echocardiographic guidance with a 7.5-mm sharp punch instrument that was forwarded via an LV incision. A special designed patch system composed of a patch with a Nitinol frame was passed across the carotid artery into the LV and positioned in front of the mVSD. An instrument resembling a stapler was introduced across the LV wall on the beating heart without use of CPB. The patch was fixed with Nitinol anchors on the septum under echocardiographic and fluoroscopic guidance. Finally, the Nitinol frame was detached from the patch. RESULTS: The locations of the defects were apical (n = 1), midmuscular (n = 3), and anterior muscular (n = 2). Closure of the mVSD was successful in 5 of 6 animals confirmed by echocardiography, hemodynamic measurements, and explantation of the heart. Animals were hemodynamically stable throughout the experiment. CONCLUSIONS: Here, we present a novel technique for hybrid closure of mVSDs without use of CPB. Further development of the patch system is necessary to assess applicability in humans, especially for the target group of newborns and infants. PMID- 18420105 TI - Antiarrhythmogenic effect of reconstituted high-density lipoprotein against ischemia/reperfusion in rats. AB - OBJECTIVES: This study analyzed the antiarrhythmogenic effect of reconstituted high-density lipoprotein (rHDL) against ischemia/reperfusion in vivo. BACKGROUND: Recent studies have suggested that a reduction in the plasma HDL level may contribute to cardiac sudden death. Although there are currently only a few therapeutic strategies for increasing HDL, an exciting new therapeutic option, rHDL, has recently been developed to prevent coronary artery disease. METHODS: To analyze the suppression of reperfusion arrhythmia by rHDL (apolipoprotein A-I with 1-palmitoyl-2-oleoyl-phosphatidyl-choline), 92 male Wistar rats were divided into 10 groups: rats that had been pre-treated with or without rHDL, apolipoprotein A-I, or 1-palmitoyl-2-oleoyl-phosphatidyl-choline in the presence or absence of inhibitors of Akt protein kinase, nitric oxide (NO), or extracellular-signal-regulated kinase (ERK) administered intravenously before left coronary artery occlusion. We also used human coronary artery endothelial cells and adenosine triphosphate-binding cassette transporter (ABC) A1-, ABCG1-, or scavenger receptor class B, type I-transfected ldlA7 cells systems. RESULTS: The duration of ventricular tachycardia or ventricular fibrillation after reperfusion in rHDL-pre-treated rats was much shorter than that in untreated rats. Apolipoprotein A-I or 1-palmitoyl-2-oleoyl-phosphatidyl-choline alone had no effect. The effect of rHDL was blocked by inhibitors of Akt, NO, and ERK. Plasma NO concentration in the rHDL group was significantly higher. In addition, rHDL activated phospho(p)-Akt, p-ERK, and p-endothelial NO synthesis in endothelial cells. The rHDL activated p-ERK in ABCA1- or ABCG1-transfected but not scavenger receptor class B, type I-transfected ldlA7 cells. CONCLUSIONS: The rHDL-induced NO production, probably mediated by ABCA1 or ABCG1 through an Akt/ERK/NO pathway in endothelial cells, may suppress reperfusion-induced arrhythmias. The HDL-based therapy may hold the promise of reducing the incidence of such arrhythmias after ischemia/reperfusion. PMID- 18420106 TI - Device interactions in patients with advanced cardiomyopathy. PMID- 18420107 TI - Safety and efficacy of remote magnetic ablation for atrial fibrillation. PMID- 18420110 TI - Right ventricular asynergy as a prognosticator. PMID- 18420111 TI - Warsaw high-preferring (WHP) and Warsaw low-preferring (WLP) lines of rats selectively bred for high and low voluntary ethanol intake: preliminary phenotypic characterization. AB - The Warsaw High Preferring (WHP) and Warsaw Low Preferring (WLP) lines were bred from Wistar foundation stock to obtain lines of rats that differ in their preference for ethanol solutions. The WHP line has met several major criteria for an animal model of alcoholism. The WHP rats voluntarily drink excessive amounts of ethanol while the WLP rats consume negligible amounts of ethanol. The WHP rats attain physiologically active blood ethanol concentrations with chronic free choice drinking. They also develop subtle but visible signs of physical dependence (the withdrawal signs). The patterns of ethanol consumption in WHP and WLP lines are stable in time and independent of the manner of access to ethanol solutions. Notably, when exposed to the increasing ethanol concentrations WHP rats gradually increased total ethanol intake whereas the WLP rats consumed invariably very low amounts of ethanol. Furthermore, the WHP rats show an increased responsiveness to the stimulatory effects of low dose of ethanol. PMID- 18420112 TI - A 3-day exposure to 10% ethanol with 10% sucrose successfully initiates ethanol self-administration. AB - The initiation phase of ethanol self-administration is difficult to study using the well-established, sucrose-fading procedure due to the changing concentrations of ethanol in the first few days. The purpose of this experiment was to test whether a modified sucrose-substitution procedure in which rats are initially exposed to high concentrations of ethanol and sucrose for three days would successfully initiate ethanol self-administration. Male Long-Evans rats were trained to lever-press with a 10% sucrose solution in which four or 20 responses allowed 20-min access to the solution. Subsequently, rats were exposed to a 3-day period of operant self-administration of 10% sucrose+10% ethanol. This constant concentration exposure was followed by the standard procedure in which sucrose is completely faded out. The establishment of ethanol self-administration was determined by ethanol intake, pre- and postprocedure two-bottle choice preference tests, and extinction trials. The mean ethanol intake was 2.2 times higher on day 2 compared with day 1 on the 10% sucrose+10% ethanol solution. After fading out the sucrose, the daily intake of 10% ethanol solution over 5 days was stable at approximately 0.57 g/kg. Ethanol preference was approximately threefold higher after the modified sucrose-fading procedure. Responding during a single session extinction test was dramatically increased from 4 to 61+/-13 or 20 to 112+/-22 responses in 20 min. Similar to the standard sucrose-fading method, we did not observe a significant relationship between extinction responding and ethanol intake. Blood alcohol concentrations were 4.5 mM 20 min after consumption began. We conclude that initiation and establishment of ethanol self-administration will occur using this modified sucrose-fading procedure. PMID- 18420115 TI - Effects of alcohol on autonomic responses and thermal sensation during cold exposure in humans. AB - We investigated the effects of alcohol on thermoregulatory responses and thermal sensations during cold exposure in humans. Eight healthy men (mean age 22.3+/-0.7 year) participated in this study. Experiments were conducted twice for each subject at a room temperature of 18 degrees C. After a 30-min resting period, the subject drank either 15% alcohol at a dose of 0.36 g/kg body weight (alcohol session) or an equal volume of distilled water (control session), and remained in a sitting position for another 60 min. Mean skin temperature continued to decrease and was similar in control and alcohol sessions. Metabolic rate was lower in the alcohol session, but the difference did not affect core temperature, which decreased in a similar manner in both alcohol and control sessions (from 36.9+/-0.1 degrees C to 36.6+/-0.1 degrees C). Whole body sensations of cold and thermal discomfort became successively stronger in the control session, whereas these sensations were both greatly diminished after drinking alcohol. In a previous study we performed in the heat, using a similar protocol, alcohol produced a definite, coordinated effect on all autonomic and sentient heat loss effectors. In the current study in the cold, as compared to responses in the heat, alcohol intake was followed by lesser alterations in autonomic effector responses, but increased changes in sensations of temperature and thermal discomfort. Overall, our results indicate that although alcohol influences thermoregulation in the cold as well as in the heat, detailed aspects of the influence are quite different. PMID- 18420114 TI - Maternal adrenocorticotropin, cortisol, and thyroid hormone responses to all three-trimester equivalent repeated binge alcohol exposure: ovine model. AB - Alcohol-mediated alterations in hypothalamic-pituitary-adrenal (HPA) and hypothalamic-pituitary-thyroid axis function are two proposed mechanisms by which alcohol causes neurodevelopmental injury to the fetus. We previously reported that third-trimester equivalent only alcohol exposure in sheep results in increases in the maternal and fetal adrenocorticotropin and cortisol levels, and decreases in the fetal thyroid hormones T(3) and T(4) and maternal T(3) levels. In this study, we wished to characterize the maternal HPA and hypothalamic pituitary-thyroid hormone responses to repeated binge alcohol exposure during all three-trimester equivalents of pregnancy in sheep. Pregnant ewes received intravenous infusions of alcohol at doses of 0.75, 1.25, or 1.75 g/kg over 1h with mean peak blood alcohol concentrations of 90, 126, or 183 mg/dl, respectively, on 3 consecutive days each week beginning on gestational day (GD) 4. Maternal blood samples were collected on GDs 6, 40, 90, and 132. Maternal plasma concentrations of adrenocorticotropin and cortisol increased in response to the high alcohol dose, and the magnitude of these elevations was not different across gestation. Thyroid hormone levels were not different when comparing among treatment groups at any time point during gestation. However, there was an ontogenetic decrease in the maternal T(3) concentration beginning between GDs 6 and 40 and a decrease in maternal T(4) and free T(4) beginning between GDs 40 and 90. The current findings suggest that (1) maternal alcohol consumption at any time during gestation stimulates the HPA axis, (2) maternal HPA responsiveness to alcohol does not change across gestation, (3) binge alcohol exposure at these doses lasting all three-trimester equivalent of human brain development does not reduce maternal thyroid hormone concentration, (4) alterations in fetal thyroid function in response to alcohol exposure do not occur as a result of diminished maternal thyroid hormone contribution, and (5) there is an ontogenetic decrease in ovine maternal thyroid hormones over gestation. PMID- 18420113 TI - Effects of the mGluR2/3 agonist LY379268 and the mGluR5 antagonist MPEP on handling-induced convulsions during ethanol withdrawal in mice. AB - In alcoholic patients, ethanol is often consumed in a repeated cyclic pattern of intoxication followed by abstinence and the emergence of withdrawal symptoms. Repeated cycles of ethanol intoxication and withdrawal lead to a sensitization of central nervous system hyperexcitability as a result of an imbalance between inhibitory GABAergic transmission and excitatory glutamatergic transmission. Symptoms of alcohol withdrawal are usually treated pharmacologically with either benzodiazepines or anticonvulsant medications. However, recent evidence suggests that inhibition of glutamate transmission by stimulation of presynaptic inhibitory metabotropic glutamate receptors (i.e., mGluR2/3 receptors) or inhibition of mGluR5 receptors produces anticonvulsant effects. Therefore, the present study was designed to determine the effects the mGluR2/3 agonist LY379268 and the mGluR5 antagonist 2-methyl-6-(phenylethynyl)-pyridine (MPEP) on ethanol withdrawal-induced seizure activity. Adult male C3H/He mice received chronic 16 h of ethanol vapor exposure in inhalation chambers followed by 8 h of withdrawal daily for 4 consecutive days. During the final (fourth) withdrawal cycle, mice were evaluated hourly for handling-induced convulsions (HIC), and were treated with vehicle, LY379268 (0.3, 1, and 3mg/kg) or MPEP (1, 3, and 10mg/kg) treatment at 4 and 8h into withdrawal. Significant reductions in overall HIC activity were not observed following administration of either compound. These results suggest that inhibition of glutamate transmission by mGluR2/3 agonists or mGluR5 antagonists does not alter HIC activity during withdrawal from repeated ethanol exposure, and as such these compounds may have limited usefulness in the treatment of central nervous system hyperexcitability during alcohol withdrawal. PMID- 18420116 TI - Diet, alcohol consumption, and serum lipid levels of the middle-aged and elderly in the Guangxi Bai Ku Yao and Han populations. AB - Bai Ku Yao is an isolated subgroup of the Yao minority in China. The special customs and cultures including their clothing, intraethnic marriages, corn wine and rum intakes are still completely conserved to the present day. Little is known about the association of diet and alcohol consumption with serum lipid levels in this population. The aim of this study was to compare the differences in diet, alcohol consumption, and serum lipid levels of the middle-aged and elderly between the Guangxi Bai Ku Yao and Han populations. A total of 485 subjects of Bai Ku Yao and 501 participants of Han Chinese aged 40 and over were surveyed by a stratified randomized cluster sampling. Information on dietary intake and alcohol consumption was collected by standard questionnaires. Serum lipid levels were measured. Education level, height, weight, body mass index, waist circumference, blood pressure, hypertension, and total energy, fat, protein, dietary cholesterol, and salt intakes were lower in Bai Ku Yao than in Han (P < .05-.001), whereas physical activity level, carbohydrate, vegetal protein, and total dietary fiber intakes were higher in Bai Ku Yao than in Han (P < .001 for all). Serum total cholesterol, high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), apolipoprotein (Apo) A1, and Apo B levels were lower in Bai Ku Yao than in Han (P < .001 for all). The levels of triglyceride, HDL-C, Apo A1, and the ratio of Apo A1 to Apo B in Bai Ku Yao were higher, but the levels of LDL-C and Apo B were lower in drinkers than in nondrinkers. The levels of triglyceride, HDL-C, LDL-C, Apo A1, Apo B, and the ratio of Apo A1 to Apo B in Bai Ku Yao were also influenced by the amount of alcohol consumed (P < .05-.001). High-density lipoprotein cholesterol levels in Han were higher and LDL-C levels were lower in drinkers than in nondrinkers (P < .01 for each). Serum total cholesterol, HDL-C, and LDL-C levels in Han were also associated with the amount of alcohol consumed (P < .05-.001). The differences in the lipid levels between the two ethnic groups may partially attribute to the differences in dietary habits and alcohol consumption. PMID- 18420117 TI - Management of thoracic trauma. Introduction. PMID- 18420118 TI - Diagnostic techniques in thoracic trauma. AB - Diagnosis of thoracic injury begins with a history of events and examination of the patient. Appropriate radiographic studies will be dictated by the findings on history and physical. Procedural examinations, such as endoscopy or angiography, may also be needed for accurate diagnosis. PMID- 18420119 TI - Critical primary survey injuries. AB - The primary survey as established by the advanced trauma life support protocol includes a directed history and physical exam aimed at the rapid diagnosis of life-threatening thoracic injuries. Most of these injuries can and should be found and treated during the initial evaluation of the patient without the need for additional diagnostic studies. In this article we review the role of the primary survey in the diagnosis and treatment of major thoracic injuries. PMID- 18420120 TI - Upper airway injury and its management. AB - Injuries to the upper airways are rare, but carry a significant morbidity and mortality. The degree of injury and presentation varies; thus recognition often requires a high index of suspicion based on mechanism. Effective management of laryngotracheal injuries begins with immediate control of the airway whether by orotracheal and surgical route. Definitive management of upper airway injuries relies on an understanding of the anatomy of the larynx, trachea and surrounding structures. Associated injuries are common and must be addressed concomitantly. Postoperative complications are frequent, requiring perioperative vigilance and long-term follow-up to ensure best outcome. PMID- 18420121 TI - Emergency department thoracotomy. AB - Emergency department thoracotomy (EDT) is defined as a thoracotomy performed in the emergency department for patients who are in extremis. The goals of the procedure are to treat pericardial tamponade, control hemorrhage, treat systemic air embolism, perform open cardiac massage and temporarily occlude the thoracic aorta. The lack of clarity in the medical literature, the need for rapid intervention in those patients deemed appropriate for the procedure, the life and death nature of the decision, and the low but finite functional survival rates following EDT for trauma, have made the conduct of this procedure a subject of great controversy among trauma experts. In this brief review, we summarize the available literature, technical concerns and indications for the procedure. We aim to provide suitable information for individual readers to refine their approach to EDT. PMID- 18420122 TI - Penetrating thoracic trauma. AB - The initial approach to penetrating thoracic trauma is directed towards the pathophysiologic syndrome upon presentation. Most patients are successfully treated with drainage tubes. The unstable patient may necessitate thoracotomy at the emergency room to drain cardiac tamponade, provide cardiac massage and control bleeding. The guidelines for this procedure are reviewed. Need for further work-up of potential injuries to other mediastinal organs is frequently screened by computerized tomography. Surgery might still be needed, on a less emergent basis, in order to repair injuries to the trachea/esophagus, retained hemothorax, or to rule out diaphragmatic injury. Laparoscopic and thoracoscopic procedures may be used in specific situations. PMID- 18420123 TI - Blunt thoracic trauma. AB - Blunt thoracic trauma represents a significant portion of trauma admissions to hospitals in the United States. These injuries are encountered by physicians in many specialities such as emergency medicine, pediatrics, general surgery and thoracic surgery. Accurate diagnosis and treatment improves the chances of favorable outcomes and it is desirable for all treating physicians to have current knowledge of all aspects of blunt thoracic trauma. Cardiothoracic surgeons often treat the most severe forms of blunt thoracic injuries and we review the aspects of blunt thoracic trauma that are pertinent to the practicing cardiothoracic surgeon. PMID- 18420124 TI - Great vessel and cardiac trauma: diagnostic and management strategies. AB - Blunt and penetrating trauma to the chest can result in great vessel or cardiac injury. Both the diagnosis and management of these thoracic injuries have evolved from more invasive to less invasive strategies paralleling the advent of sophisticated imaging tools and the development of endovascular therapies. Despite these advances, conventional open repair and reconstruction techniques remain important and are often the definitive means toward effective management of these severely injured patients. The following review outlines the historical perspective, diagnosis, and management of blunt thoracic aortic, blunt cardiac, and penetrating cardiac injuries. PMID- 18420125 TI - Flail chest and pulmonary contusion. AB - Flail chest is most often accompanied by a significant underlying pulmonary parenchymal injury and can be a life-threatening thoracic injury. Its management is often complicated by the other injuries it is frequently associated with. Similarly, mortality and morbidity are dictated most often by the associated injuries and findings. Its treatment is complex and should first be one of pain management, judicious fluid resuscitation, and excellent pulmonary toilet. In those patients requiring mechanical ventilatory support, or who require ipsilateral thoracocotomy, rib stabilization may be considered depending on a host of potentially conflicting indications and contraindications. At the end of this section are listed the current major recommendations and their levels of evidence. PMID- 18420126 TI - Esophageal trauma. AB - The anatomy of the esophagus is unique in that it traverses the neck, chest, and abdomen. As a result, surgeons need to be familiar with the anatomy of all three of these areas to be facile and comfortable in performing esophageal surgery. Traumatic injuries to the esophagus encompass a heterogeneous group of injuries that can be iatrogenic, external, or from physiologic forces. Primary repair of traumatic injuries is preferred when possible; however, if systemic sepsis is present and esophageal resection becomes necessary due to extensive injury or inflammation, immediate reconstruction should be delayed in most cases. Successful management of traumatic esophageal injuries requires prompt and accurate diagnosis and treatment tailored specifically to both the type of injury as well as to the patient's overall clinical condition. PMID- 18420127 TI - Tracheobronchial injury. AB - Tracheobronchial injuries (TBI) can be challenging to diagnose, manage, and definitively treat. They encompass a heterogeneous group of injuries that are often associated with other injuries. Although relatively rare, diagnosis and treatment of TBI often requires skillful and creative airway management, careful diagnostic evaluation, and operative repairs that are often resourceful and necessarily unique to the given injury. An experienced surgeon with a high level of suspicion and the liberal use of bronchoscopy constitute the major tools necessary for diagnosing and treating these injuries. Most TBI can be repaired primarily using a tailored surgical approach and techniques specific to the injury. Associated injuries are common, and surgeons must be knowledgeable in treating a wide variety of physiologic abnormalities, especially those involving the chest wall and lung parenchyma, if a successful outcome is to be achieved in the management of these often challenging patients. PMID- 18420128 TI - The pediatric patient and thoracic trauma. AB - Trauma remains the leading cause of death for children aged 1 to 14 years. Thoracic trauma is seen in 4% to 6% of pediatric patients presenting to pediatric trauma centers and rarely occurs in isolation. The medical and surgical evaluation of children is a challenging task to even the most experienced physician. Effective treatment of the pediatric trauma patient can only be provided if the physician understands the major pitfalls which are common in the pediatric population. The assessment of the pediatric patient is simplified by an understanding of specific anatomic and physiologic differences between children and adults. While noting children are not small adults, the systematic approach taken towards the evaluation of an adult is similar. Sequential evaluation and management of the ABCs by a caregiver familiar with age specific norms is the most important initial consideration. The care of specific injuries is similar to those found in adults but the patient's size limits the physician's options in many cases. PMID- 18420129 TI - The multiply injured patient. AB - The multiply injured patient with significant thoracic and extra-thoracic injuries poses a number of challenges. Pericardial tamponade, tension pneumothorax and massive hemothorax can and should be diagnosed clinically. In more stable patients, chest computed tomography (CT) scan is an excellent screening test. The concept of damage control resuscitation and damage control surgery have shown promise in patients with multiple, critical injuries. Beta blockade of patients with blunt thoracic aortic injuries can be used as a temporizing damage control measure when the risks of operation or intervention are very high (traumatic brain injury, severe right or bilateral pulmonary contusion, unstable pelvic fractures). Patients with multiple penetrating wounds require the surgical team to be expeditious and flexible, and damage control is a helpful strategy in these patients. PMID- 18420130 TI - Common complications following thoracic trauma: their prevention and treatment. AB - Although there are a wide range of complications following thoracic trauma, respiratory failure, pneumonia, and pleural sepsis are the most common potentially preventable problems. Respiratory failure and pneumonia are directly related to the severity of the injury and the age and condition of the patient. A program aimed at aggressive pain control, mobilization, and pulmonary care can reduce the risk of respiratory failure, pneumonia, and death in these patients. Pleural sepsis develops in the face of a retained hemothorax, which becomes contaminated with bacteria. The most common source for this contamination is not pneumonia, but external contamination from the wound itself or at the time of placement of the tube thoracostomy. Measures that reduce the volume of retained pleural blood and reduce or eliminate any bacterial contamination are likely to reduce the incidence of pleural sepsis. The authors review these complications and describe a plan to reduce these complications. PMID- 18420131 TI - Minimally invasive techniques in thoracic trauma. AB - Endovascular techniques and videoscopic assisted thoracoscopic surgery have been selectively applied in the trauma setting. These techniques continue to evolve and have gained acceptance as the treatment of choice for certain traumatic thoracic injuries. PMID- 18420132 TI - Combat casualty care in an air force theater hospital: perspectives of recently deployed cardiothoracic surgeons. AB - Current military operations have generated a large number of casualties and have led to the establishment of the first Air Force Theater Hospital since Vietnam. Located at Balad Airbase, Iraq, this hospital is a busy trauma center. Thoracic injuries are relatively infrequent but highly lethal. The cardiothoracic surgeon is uniquely trained to provide sophisticated surgical management to some of the most severely injured patients. The operative experiences of four recently deployed cardiothoracic surgeons are described. Mortality from combat injury in this conflict is lower than in prior wars. Body armor may prevent some fatal injuries. Several features of military medical care process are helping to improve our outcomes-specifically, the development of a trauma care system modeled on successful civilian centers, the expanded use of damage control concepts, and utilization of early transportation out of the theater of operations using Critical Care Air Transport Teams (CCATT). PMID- 18420133 TI - Regeneron focuses on age-related macular degeneration. PMID- 18420134 TI - PAK'n it in: identification of a selective PAK inhibitor. AB - p21-activated kinases are important signaling kinases for which no specific chemical inhibitors are known. In this issue of Chemistry & Biology, Deacon et al. target allosteric transitions undergone during PAK activation to identify a selective inhibitor (Deacon et al., 2008). PMID- 18420135 TI - Sweetly expanding enzymatic glycodiversification. AB - Directed evolution is a powerful tool to modify substrate specificity for a wide array of enzyme catalysts. In this issue of Chemistry & Biology, Thorson and coworkers use directed evolution to increase the catalytic proficiency of a model glycosyltransferase, OleD, 300-fold for a nonphysiological substrate (Williams et al., 2008). PMID- 18420136 TI - The tail of mycolic acids. AB - The FabH enzyme from M. tuberculosis binds the acyl tail of large substrates at the end of a buried hydrophobic tunnel. Sachdeva et al. (2008) use reactive chemical probes and X-ray crystallography to show that substrates can bind to an open state of FabH without threading through the tunnel. PMID- 18420137 TI - Studying posttranslational modifications by in-cell NMR. AB - Functional in vivo investigation of posttranslational modifications is a problem that a number of analytical techniques are trying to tackle. Below, we briefly discuss the breakthroughs and challenges in placing NMR spectroscopy on the map, as illustrated by a recent report by Selenko et al. (2008). PMID- 18420138 TI - Ethylene receptor antagonists: strained alkenes are necessary but not sufficient. AB - Plants use ethylene as a hormone to control many physiological processes. Ethylene perception involves its binding to an unusual copper-containing, membrane-bound receptor. Inhibitors of ethylene action are valuable to study signaling and may have practical use in horticulture. Past investigation of alkene ligands for this receptor has identified strain as the key factor in antagonism of ethylene binding and action, consistent with known trends in metal alkene complex stability. However, in this work, this principle could not be extended to other alkenes, prompting development of the proposal that a ring opening reaction accounts for the unusual potency of cyclopropene ethylene antagonists. Another factor augmenting the affinity of alkenes for the copper binding site is pyramidalization, as in trans-cycloalkenes. The enantiomeric selectivity in the binding of one such alkene to the ethylene receptor demonstrates its protein-composed asymmetric environment. PMID- 18420139 TI - An isoform-selective, small-molecule inhibitor targets the autoregulatory mechanism of p21-activated kinase. AB - Autoregulatory domains found within kinases may provide more unique targets for chemical inhibitors than the conserved ATP-binding pocket targeted by most inhibitors. The kinase Pak1 contains an autoinhibitory domain that suppresses the catalytic activity of its kinase domain. Pak1 activators relieve this autoinhibition and initiate conformational rearrangements and autophosphorylation events leading to kinase activation. We developed a screen for allosteric inhibitors targeting Pak1 activation and identified the inhibitor IPA-3. Remarkably, preactivated Pak1 is resistant to IPA-3. IPA-3 also inhibits activation of related Pak isoforms regulated by autoinhibition, but not more distantly related Paks, nor >200 other kinases tested. Pak1 inhibition by IPA-3 in live cells supports a critical role for Pak in PDGF-stimulated Erk activation. These studies illustrate an alternative strategy for kinase inhibition and introduce a highly selective, cell-permeable chemical inhibitor of Pak. PMID- 18420140 TI - Solvent structure and hammerhead ribozyme catalysis. AB - Although the hammerhead ribozyme is regarded as a prototype for understanding RNA catalysis, the mechanistic roles of associated metal ions and water molecules in the cleavage reaction remain controversial. We have investigated the catalytic potential of observed divalent metal ions and water molecules bound to a 2 A structure of the full-length hammerhead ribozyme by using X-ray crystallography in combination with molecular dynamics simulations. A single Mn(2+) is observed to bind directly to the A9 phosphate in the active site, accompanying a hydrogen bond network involving a well-ordered water molecule spanning N1 of G12 (the general base) and 2'-O of G8 (previously implicated in general acid catalysis) that we propose, based on molecular dynamics calculations, facilitates proton transfer in the cleavage reaction. Phosphate-bridging metal interactions and other mechanistic hypotheses are also tested with this approach. PMID- 18420141 TI - Toward deciphering the code to aminergic G protein-coupled receptor drug design. AB - The trace amine-associated receptor 1 (TAAR(1)) is a biogenic amine G protein coupled receptor (GPCR) that is potently activated by 3-iodothyronamine (1, T(1)AM) in vitro. Compound 1 is an endogenous derivative of the thyroid hormone thyroxine which rapidly induces hypothermia, anergia, and bradycardia when administered to mice. To explore the role of TAAR(1) in mediating the effects of 1, we rationally designed and synthesized rat TAAR(1) superagonists and lead antagonists using the rotamer toggle switch model of aminergic GPCR activation. The functional activity of a ligand is proposed to be correlated to its probable interactions with the rotamer switch residues; agonists allow the rotamer switch residues to toggle to their active conformation, whereas antagonists interfere with this conformational transition. These agonist and antagonist design principles provide a conceptual model for understanding the relationship between the molecular structure of a drug and its pharmacological properties. PMID- 18420142 TI - Structure-activity relationships of antibacterial acyl-lysine oligomers. AB - We describe structure-activity relationships that emerged from biophysical data obtained with a library of antimicrobial peptide mimetics composed of 103 oligoacyllysines (OAKs) designed to pin down the importance of hydrophobicity (H) and charge (Q). Based on results obtained with OAKs displaying minimal inhibitory concentration < or = 3 microM, the data indicate that potent inhibitory activity of the gram-negative Escherichia coli and the gram-positive Staphylococcus aureus required a relatively narrow yet distinct window of HQ values where the acyl length played multiple and critical roles, both in molecular organization and in selective activity. Thus, incorporation of long-but not short-acyl chains within a peptide backbone is shown to lead to rigid supramolecular organization responsible for poor antibacterial activity and enhanced hemolytic activity. However, sequence manipulations, including introduction of a tandem lysine motif into the oligomer backbone, enabled disassembly of aggregated OAKs and subsequently revealed tiny, nonhemolytic, yet potent antibacterial derivatives. PMID- 18420144 TI - Mitochondria-penetrating peptides. AB - Mitochondria are important targets for cancer chemotherapy and other disease treatments. Gaining access to this organelle can be difficult, as the inner membrane is a barrier limiting diffusive transport. A mitochondrial molecular carrier would be a boon to the development of organelle-specific therapeutics. Here, we report a significant advance in the development of mitochondrial transporters-synthetic cell-permeable peptides that are able to enter mitochondria. Efficient uptake of these mitochondria-penetrating peptides (MPPs) is observed in a variety of cell types, and organellar specificity is attained with sequences that possess specific chemical properties. The MPPs identified are cationic, but also lipophilic; this combination of characteristics facilitates permeation of the hydrophobic mitochondrial membrane. The examination of a panel of MPPs illustrates that mitochondrial localization can be rationally controlled and finely tuned by altering lipophilicity and charge. PMID- 18420143 TI - PAP inhibitor with in vivo efficacy identified by Candida albicans genetic profiling of natural products. AB - Natural products provide an unparalleled source of chemical scaffolds with diverse biological activities and have profoundly impacted antimicrobial drug discovery. To further explore the full potential of their chemical diversity, we survey natural products for antifungal, target-specific inhibitors by using a chemical-genetic approach adapted to the human fungal pathogen Candida albicans and demonstrate that natural-product fermentation extracts can be mechanistically annotated according to heterozygote strain responses. Applying this approach, we report the discovery and characterization of a natural product, parnafungin, which we demonstrate, by both biochemical and genetic means, to inhibit poly(A) polymerase. Parnafungin displays potent and broad spectrum activity against diverse, clinically relevant fungal pathogens and reduces fungal burden in a murine model of disseminated candidiasis. Thus, mechanism-of-action determination of crude fermentation extracts by chemical-genetic profiling brings a powerful strategy to natural-product-based drug discovery. PMID- 18420145 TI - Structure-based design of a superagonist ligand for the vitamin D nuclear receptor. AB - Vitamin D nuclear receptor (VDR), a ligand-dependent transcriptional regulator, is an important target for multiple clinical applications, such as osteoporosis and cancer. Since exacerbated increase of calcium serum level is currently associated with VDR ligands action, superagonists with low calcium serum levels have been developed. Based on the crystal structures of human VDR (hVDR) bound to 1alpha,25-dihydroxyvitamin D(3) and superagonists-notably, KH1060-we designed a superagonist ligand. In order to optimize the aliphatic side chain conformation with a subsequent entropy benefit, we incorporated an oxolane ring and generated two stereo diasteromers, AMCR277A and AMCR277B. Only AMCR277A exhibits superagonist activity in vitro, but is as calcemic in vivo as the natural ligand. The crystal structures of the complexes between the ligand binding domain of hVDR and these ligands provide a rational approach to the design of more potent superagonist ligands for potential clinical application. PMID- 18420148 TI - Neuroblastoma screening: investing in the right test. PMID- 18420146 TI - Optimizing glycosyltransferase specificity via "hot spot" saturation mutagenesis presents a catalyst for novobiocin glycorandomization. AB - A comprehensive two-phase "hot spot" saturation mutagenesis strategy for the rapid evolution of glycosyltransferase (GT) specificity for nonnatural acceptors is described. Specifically, the application of a high-throughput screen (based on the fluorescent acceptor umbelliferone) was used to identify key amino acid hot spots that contribute to GT proficiency and/or promiscuity. Saturation mutagenesis of the corresponding hot spots facilitated the utilization of a lower throughput screen to provide OleD prodigy capable of efficiently glycosylating the nonnatural acceptor novobiocic acid with an array of unique sugars. Incredibly, even in the absence of a high-throughput screen for novobiocic acid glycosylation, this approach rapidly led to improvements in the desired catalytic activity of several hundred-fold. PMID- 18420147 TI - Separate entrance and exit portals for ligand traffic in Mycobacterium tuberculosis FabH. AB - Mycobacterium tuberculosis FabH initiates type II fatty acid synthase-catalyzed formation of the long chain (C(16)-C(22)) acyl-coenzyme A (CoA) precursors of mycolic acids, which are major constituents of the bacterial cell envelope. Crystal structures of M. tuberculosis FabH (mtFabH) show the substrate binding site to be a buried, extended L-shaped channel with only a single solvent access portal. Entrance of an acyl-CoA substrate through the solvent portal would require energetically unfavorable reptational threading of the substrate to its reactive position. Using a class of FabH inhibitors, we have tested an alternative hypothesis that FabH exists in an "open" form during substrate binding and product release, and a "closed" form in which catalysis and intermediate steps occur. This hypothesis is supported by mass spectrometric analysis of the product profile and crystal structures of complexes of mtFabH with these inhibitors. PMID- 18420149 TI - Preterm birth and the developing brain. PMID- 18420150 TI - A heterozygous frameshift mutation in PRKRA (DYT16) associated with generalised dystonia in a German patient. PMID- 18420152 TI - Modelling of neuronal circuits: the future is bright. PMID- 18420153 TI - Profile: Kenneth Rockwood: on a mission to save the world? PMID- 18420155 TI - The industrialisation of neuroscience. PMID- 18420156 TI - Recent developments in childhood arterial ischaemic stroke. AB - Stroke is increasingly recognised as a cause of childhood disability and lifelong morbidity: population-based estimates of the annual incidence of childhood stroke (ischaemic and haemorrhagic) range from 2.3 to 13.0 per 100,000 children, and incidence rates in neonates are closer to 1 per 5000 livebirths. Stroke in childhood can have many causes. Diagnosis is often delayed owing to low clinical suspicion and the need to exclude the frequent mimics of stroke in childhood. Outcomes are related to presentation, associated illnesses, the underlying cause, size, and location of the infarct, and stroke subtype, but more than a half of the children who have had a stroke will have long-term neurological sequelae. Furthermore, estimates of recurrence rates range from 6-19% in the first few years. Arteriopathy-including arterial dissection and other progressive and non progressive arteriopathies-might account for up to 80% of childhood stroke in otherwise healthy children. Because children with cerebrovascular abnormalities are at the highest risk of recurrence (66% at 5 years), understanding of the nature and course of these arteriopathies is crucial to the development of secondary stroke prevention strategies. PMID- 18420157 TI - Endpoints for trials in Alzheimer's disease: a European task force consensus. AB - Harmful consequences in health status caused by disease are referred to as outcomes, and in clinical studies the measures of these outcomes are called endpoints. A major challenge when deciding on endpoints is to represent the outcomes of interest accurately, and the accuracy of such representation is assessed through validation. Complex diseases like Alzheimer's disease have many different and interdependent outcomes. We present a consensus for endpoints to be used in clinical trials in Alzheimer's disease, agreed by a European task force under the auspices of the European Alzheimer Disease Consortium. We suggest suitable endpoints for primary and secondary prevention trials, for symptomatic and disease-modifying trials in very early, mild, and moderate Alzheimer's disease, and for trials in severe Alzheimer's disease. A clear and consensual definition of endpoints is crucial for the success of further clinical trials in the field and will allow comparison of data across studies. PMID- 18420159 TI - Metastatic epidural spinal cord compression. AB - Metastatic epidural spinal cord compression (MESCC) occurs when cancer metastasises to the spine or epidural space and causes secondary compression of the spinal cord. MESCC is a common complication of malignancy that affects almost 5% of patients with cancer. The most common symptom is back pain. MESCC is a medical emergency that needs rapid diagnosis and treatment if permanent paralysis is to be prevented: the diagnosis of MESCC is best made with MRI; and corticosteroids, radiation therapy, and surgery are all established treatments. Future research will focus on prevention, improving detection, and the development of new treatments. PMID- 18420160 TI - Sex and society: the journal takes a look at their interaction. PMID- 18420158 TI - Management of neurogenic orthostatic hypotension: an update. AB - Orthostatic hypotension (OH) is common in elderly people and in patients with disorders such as diabetes and Parkinson's disease. Grading of the severity of OH and its effect on the patient's quality of life are important. The symptoms vary with orthostatic stress, and subtle symptoms such as tiredness and cognitive impairment should be recognised. Standard drug treatment for OH is effective but worsens supine hypertension, whereas pyridostigmine can improve OH slightly but significantly without worsening of supine hypertension. Because orthostatic stress varies from moment to moment and drug treatment is suboptimal, drug treatment of OH needs to be combined with non-pharmacological approaches, such as compression of venous capacitance beds, use of physical counter-manoeuvres, and intermittent water-bolus treatment. PMID- 18420161 TI - Embryonic stem cells in science and medicine, part II: law, ethics, and the continuing need for dialogue. AB - Just as our first article, "Embryonic Stem Cells in Science and Medicine: An Invitation for Dialogue," in the December 2007 issue of Gender Medicine went to press, two groups of researchers had just announced that adult human somatic cells had been reprogrammed to behave like pluripotent stem cells, and that the reprogrammed cells were able to differentiate into cell types of the 3 germ layers in vitro and in a mouse model. A third group has since done so. Because the reprogrammed cells were not embryonic in origin, the announcements were heralded as "stunning" and "leaps forward," because, it was argued, the ability to generate stem cells, without destroying embryos in the process, would avoid the difficult ethical questions raised by human embryonic stem (hES) cell research. This article addresses the most recent announcements and briefly retraces the relevant history so that we may consider whether the moral, ethical, and social issues do in fact disappear as a result of these new advancements. We conclude that, despite the hoopla, little has changed. If indeed there were ethical issues surrounding hES cell research, they remain-and remain as urgent to address and resolve as they had been previously. Lastly, we argue that the medical and scientific communities continue to do themselves a disservice by failing to create a cohesive governing body to address and make concrete recommendations concerning the moral, ethical, and related social issues affecting their communities. PMID- 18420162 TI - Mechanisms underlying sex differences in progressive renal disease. AB - Men with nondiabetic renal disease exhibit a faster rate of decline in renal function compared with women. To investigate this sex difference in renal disease progression, our research group has been studying the renal wrap (RW) model of hypertension in rats. Compared with RW female rats, the glomerulosclerosis index, mean glomerular volume, and proteinuria were greater (3.1-, 1.7-, and 1.8-fold, respectively) in RW males under conditions in which no differences in the degree of hypertension were detected, suggesting that sex differences may exist in the mechanisms underlying renal injury, independent of blood pressure. Gonadal steroids contribute to these sex differences, because orchidectomy attenuated and ovariectomy exacerbated the severity of renal injury, whereas dihydrotestosterone and 17beta-estradiol (E(2)) replacement prevented these respective effects. Chronic renal disease is associated with impairment in nitric oxide (NO) signaling and elevated levels of superoxide. Sex differences were observed in RW induced changes in renal nitric oxide synthesis (NOS) protein abundance. Whereas RW had no effect on NOS in the female kidney, endothelial NOS was elevated and neuronal NOS was decreased in the male kidney, suggesting that renal injury may cause dysfunction in NO metabolism in the male. Sex differences in superoxide signaling were also observed. Renal cortical nicotinamide adenine dinucleotide phosphate oxidase activity was 1.3-fold higher in RW males than in RW females, and ovariectomy increased enzyme activity 1.4-fold, whereas E(2) replacement prevented this effect. These changes in enzyme activity were mirrored by changes in protein abundance of the p22(phox) regulatory subunit. Our findings suggest that E(2) may protect the female kidney from hypertension-associated renal disease by attenuating injury-induced superoxide production. PMID- 18420163 TI - Gender differences in methamphetamine use and responses: a review. AB - BACKGROUND: Men and women differ markedly with regard to their use of, and responses to, methamphetamine (MA) and related amphetamines. However, these gender differences oftentimes are given only a cursory consideration in the analyses of these MA effects. OBJECTIVE: In this brief review, we summarize the data on gender differences in various parameters of MA use and responses. Such information on the pattern of male versus female differences in the use and responses to this psychostimulant can aid in tailoring gender-dependent treatment strategies. METHODS: English-language articles were identified from MEDLINE as well as from reference lists of identified articles for the years 1966 to 2007. Search terms included various combinations of men/male, women/female, methamphetamine, and gender/sex differences. Only studies with human subjects were reviewed. RESULTS: Women tend to begin MA use at earlier ages, appear more dependent on MA, but also respond better to treatment than do men. MA use appears to be associated with depression in women, and women seem more committed to MA, whereas men are more likely to use other drugs in the absence of access to MA. Female MA abusers had both larger volumes within the corpus callosum and more hyperperfused regions in the parietal and occipital areas of the brain, along with more genetic alterations but less MA-induced toxicity. Amphetamine stimulated dopamine release was greater in men. CONCLUSIONS: When considered in total, women seem more dependent on and committed to MA but show diminished (amphetamine-stimulated) dopamine responses and a decreased degree of toxicity, as indicated by a lower incidence of emergency department-related deaths involving MA. A pervasive comorbidity of depression or depression-related characteristics were present in women MA users, suggesting that MA may serve as a type of self-medication for their depression. These findings not only highlight the need for consideration of gender when assessing MA use, but also can serve to direct efforts at prevention and treatment programs that address the specific needs of men and women. PMID- 18420164 TI - Current issues in the treatment of women with abdominal aortic aneurysm. AB - BACKGROUND: Abdominal aortic aneurysm (AAA) accounts for approximately 45,000 deaths per year in the United States. Despite a striking male predominance of AAA (4:1 male to female), mortality from this disease is almost as high in women (20th leading killer of women and 15th leading killer of men in this country). OBJECTIVE: The purpose of this review is to highlight the differences in diagnosis, treatment, and treatment outcomes for women with AAA to determine avenues of potential improvement in their care. METHODS: Published articles relevant to this review were determined by the experience of the author, by PubMed and MEDLINE searches, and by reviewing the references cited in the reports identified by the first 2 methods. The database searches were performed using the following terms: abdominal aorta, aneurysm, gender, endovascular, and outcomes. Reports were limited to the English language and publication since 1995. RESULTS: Compared with men, women are older when their AAA is diagnosed and treated. Women have higher mortality than do men while undergoing elective open and endovascular repairs, and emergency surgery for ruptured AAAs. Owing to the anatomic complexity of their arterial anatomy, women are less frequently candidates for endovascular repair. Women receive treatment for rupture of AAA less frequently than do men. On Medicare induction, both men and women are eligible for a one time screening for AAA; however, women qualify for this exam only if they have a family history of AAA. CONCLUSIONS: Opportunities to advance the care of women with AAA include improving screening techniques to find AAA prior to rupture and when women are younger and more likely to be candidates for repair. Current clinical practice should focus on decreasing mortality for open surgical repair and developing better endovascular devices so that anatomic obstacles can be overcome and more women can be candidates for this technology. In addition, furthering the understanding of gender differences in the pathophysiology of AAA disease may provide insights into treatments that could prevent the formation of aneurysms. PMID- 18420165 TI - Pilot study of sex differences in chemokine/cytokine markers of atherosclerosis in humans. AB - BACKGROUND: Atherogenic processes increase in women after menopause, when the risk of cardiovascular adverse events approaches that observed in age-matched men. In experimental animals, ovariectomy increases the platelet content of mitogenic cytokines, such as platelet-derived growth factor (PDGF), which when released into the blood or site of vascular injury, contribute to atherogenic processes. OBJECTIVE: Experiments were designed to assess the sex distribution of inflammatorychemokines/cytokines, which may be released from platelets in the serum of middle-aged women and men in whom the extent of atherosclerotic coronary disease was defined by coronary arterial calcification (CAC). METHODS: Blood was obtained from healthy white individuals recruited from the Mayo Clinic database. CAC was assessed by 64-slice computed tomography. Plasma cholesterol, lipids, and high-sensitivity C-reactive protein were analyzed by the Mayo Clinic Department of Laboratory Medicine and Pathology. Serum cytokines were determined using cytokine arrays. Cytokine expression was measured using dot blot analysis. RESULTS: Of the 16 individuals (11 women, 5 men) who agreed to participate in the study, 1 woman was premenopausal, 1 was taking oral contraceptives, and 1 was receiving menopausal hormone therapy. One woman had an active infection and was eliminated from the study. CAC was detected in only 2 of the 11 women (scores of 46 and 56 Agatston units [AU]) but in 3 of the 5 men (scores of 3, 123, and 609 AU). Correcting for all other risk factors, expression of the chemokine RANTES (regulated on activation, normal, T-cell expressed and secreted; CCL5 [CC chemokine ligand 5]) was 100.98% greater in women than in men, and PDGF-BB was 55.30% greater in women than in men. CONCLUSIONS: This small pilot study found that the circulating chemokines/cytokines RANTES and PDGF-BB showed sex-disparate distribution between the women and men studied, and did not appear related to the degree of CAC. PMID- 18420166 TI - Women tolerate drug therapy for coronary artery disease as well as men do, but are treated less frequently with aspirin, beta-blockers, or statins. AB - BACKGROUND: Women have worse morbidity, mortality, and health-related quality-of life outcomes associated with coronary artery disease (CAD) compared with men. This may be related to underutilization of drug therapies, such as aspirin, beta blockers, angiotensin-converting enzyme (ACE) inhibitors, or statins. No studies have sought to describe the relationship of gender with adverse reactions to drug therapy (ADRs) for CAD in clinical practice. OBJECTIVE: The aim of this study was to determine the prevalence of ADRs associated with common CAD drug therapies in women and men in clinical practice. METHODS: In a cohort of consecutive outpatients with CAD, detailed chart abstraction was performed to determine the use of aspirin, beta-blocker, ACE inhibitor, and statin therapy, as well as the ADRs reported for these treatments. Baseline clinical characteristics were also determined to identify the independent association of gender with use of standard drug treatments for CAD. RESULTS: Consecutive patients with CAD (153 men, 151 women) were included in the study. Women and men were observed to have a similar prevalence of cardiac risk factors and comorbidities, except that men had significantly higher prevalence of atrial fibrillation (30 [19.6%] men vs 15 [9.9%] women; P = 0.03) and significantly lower mean (SD) high-density lipoprotein cholesterol concentrations (45 [16] mg/dL for men vs 55 [19] mg/dL for women; P < 0.001). No significant differences were observed between the sexes in the prevalence of ADRs; however, significantly fewer women than men were treated with statins (118 [78.1%] vs 139 [90.8%], respectively; P = 0.003). After adjusting for clinical characteristics, women were also found to be less likely than men to receive aspirin (odds ratio [OR] = 0.164; 95% CI, 0.083-0.322; P = 0.001) and beta-blockers (OR = 0.184; 95% CI, 0.096-0.351; P = 0.001). CONCLUSIONS: Women and men experienced a similar prevalence of ADRs in the treatment of CAD; however, women were significantly less likely to be treated with aspirin, beta-blockers, and statins than were their male counterparts. To optimize care for women with CAD, further study is needed to identify the cause of this gender disparity in therapeutic drug use. PMID- 18420167 TI - Functional dyspepsia affects women more than men in daily life: a case-control study in primary care. AB - BACKGROUND: Little is known about possible gender differences among patients with functional dyspepsia (FD). Few studies have measured health-related quality of life (HRQoL) in patients with FD using a population-based control group as a reference. OBJECTIVES: This study aimed to determine the degree of HRQoL impairment among patients with FD, to assess the self-reported health impact resulting from the disease, and to analyze any gender differences. METHODS: A questionnaire that included the HRQoL Short Form 36 (SF-36) Health Survey, Hospital Anxiety and Depression Scale, and other measurements was mailed to patients with FD identified from medical records. The control group was randomly selected from the general population in the same geographical area. Responses to the SF-36 were transferred to a standard scale ranging from 0 (the worst possible score) to 100 (the best possible score). RESULTS: Responders were assigned to 2 gender-specific subgroups, each with 88 patients with FD and 344 randomly matched controls, all aged 18 to 65 years. Compared with the controls, the HRQoL of patients with FD was impaired in all SF-36 dimensions except one -- role limitations caused by emotional problems. Female patients with FD had a significantly lower SF-36 score in the physical functioning dimension than did male patients (82.4 vs 90.5, respectively; P < 0.01). Both groups of patients with FD had impaired HRQoL compared with their respective control group in the dimensions of bodily pain (women: 69.3 vs 80.6, P < 0.001; and men: 75.8 vs 84.8, P < 0.001) and general health (women: 62.0 vs 75.6, P < 0.001; and men: 70.6 vs 78.6, P < 0.001). Additionally, women with FD had significant impairment compared with their respective control group in the dimensions of physical functioning (82.4 vs 89.3; P < 0.01) and physical role limitations (72.1 vs 85.9; P < 0.001). Depression was significantly more common among male patients with FD than among male controls (6.8% vs 2.0%, respectively; P < 0.05). More gastrointestinal comorbidity was reported among patients of both sexes compared with controls. CONCLUSIONS: This population-based case-control study reported HRQoL impairment overall among patients with FD. This impairment was more apparent in female patients than in female controls. Females with FD tended to be more negatively affected in their daily life than their male counterparts. These gender differences should be investigated further in future studies. PMID- 18420168 TI - Estrogen effects on MMP-13 and MMP-14 regulation of left ventricular mass in Dahl salt-induced hypertension. AB - BACKGROUND: Female Dahl salt-sensitive (DS) rats fed a low-salt diet develop hypertension at 6 months of age. Ovariectomy at 2 months of age accelerates the development of hypertension, and estrogen replacement delays it. Although acute pressure overload induces structural changes in the left ventricle (LV) further effects of gradual hypertension on LV remodeling have not been examined in the DS rat model. OBJECTIVE: The purpose of this study was to test the hypothesis that aging and estrogen loss in hypertensive DS rats are accompanied by changes in LV remodeling. METHODS: Four groups of DS rats were examined: young intact, middle aged (MA) intact, MA ovariectomized (MA-OVX), and MA-OVX with 17beta-eestradiol (E(2)) supplementation (MA-OVX+E(2)). Myocardial matrix metalloproteinases (MMPs),tissue inhibitors of metalloproteinases (TIMPs),and extracellular matrix (ECM) proteins were assessed by immunoblotting. RESULTS: Each of the 4 groups comprised 6 animals. Mean (SEM) LV mass was significantly greater in the MA intact and the MA-OVX groups (1257 [31] mg and 1199 [25] mg, respectively; both, P < 0.05) compared with the young-intact group (697 [6] mg). LV mass in the MA OVX+E(2) group was significantly lower compared with the MA-intact and MA-OVX groups (both, P < 0.05), suggesting that estrogen may attenuate LV remodeling. Fibronectin and collagen III and IV concentrations increased significantly in the MA-intact and MA-OOVX groups (all, P < 0.05),indicating increased fibrosis. Multiple MMPs also increased in the MA-intact an nd MA-OVX rats, including MMP-3, -7, -99, -113, and -114, and all TIMPs. In contrast, estrogen attenuated fibrosis by increasing MMP-8 concentrations and increasing collagen III fragments. From good-fit regression modeling, MMP-13 and MMP-14 concentrations correlated positively with LV mass for the MA-intact and MA-OVX groups, respectively. CONCLUSIONS: Gradual hypertension stimulated ECM turnover by increasing both MMP/TIMP production and ECM degradation. Estrogen loss or gain resulted in a shift in MMP profiles, suggesting that MMP-13 and MMP-14 may be differentially regulated in postmenopausal hypertension. PMID- 18420169 TI - Whack! I've hit the glass ceiling! Women's efforts to gain status in surgery. PMID- 18420170 TI - Gender imbalance in kidney transplantation: Iran in a global perspective. PMID- 18420171 TI - [Nephrologie & Therapeutique: what is new in 2008?]. PMID- 18420172 TI - Hepatitis D and B virus genotypes in chronically infected patients from the Eastern Amazon Basin. AB - Hepatitis D virus (HDV) is a defective hepatotropic virus whose infectivity is dependent on hepatitis B virus (HBV). HDV super- or co-infection leads to an increased risk of fulminant hepatitis or progression to severe chronic liver disease in HBV infected patients. The Brazilian Amazon Basin has been reported to be endemic for HBV and HDV, especially in the Western Amazon Basin. In this region, HDV infection is frequently associated with acute fulminant hepatitis with characteristic histologic features. HDV is classified into seven major clades (HDV-1 to HDV-7) and HBV is subdivided into eight genotypes (A-H). HDV and HBV genotypes have been shown to have a distinct geographic distribution. The aim of this study was to determine the HBV and HDV genotypes harbored by chronically infected patients from the Eastern Amazon Basin, Brazil. We studied 17 serum samples from HBV and HDV chronically infected patients admitted to a large public hospital (Santa Casa de Misericordia) at Belem, state of Para, Brazil, between 1994 and 2002. HDV-3 and HBV genotype A (subtype adw2) have been identified in all cases, in contrast to previous studies from other regions of the Amazon, where HBV genotype F has been found co-infecting patients that harbored HDV-3. The HDV-3/HBV-A co-infection suggests that there is not a specific interaction between HBV and HDV genotypes, and co-infection might merely reflect the most frequent genotypes found in a particular geographic area. The analysis of the carboxy-terminal region of the large hepatitis D antigen (L-HDAg), which interacts with the hepatitis B surface antigen (HBsAg) and is essential for HDV assembly, showed some diversity between the different isolates from the Eastern Amazon. This diversity is not observed among HDV-3 sequences from other South American regions. PMID- 18420173 TI - Oxidative stress induced changes in plasma protein can be a predictor of imminent severe dengue infection. AB - OBJECTIVES: Oxidative stress in dengue viral infection has been suggested and severity of it was found to be associated with progress of illness. Hence assessing oxidative stress mediated changes in plasma proteins can be an early biomarker for prediction of severe dengue infection. DESIGN AND METHODS: Thirty two dengue hemorrhagic fever (DHF), 21 dengue shock syndrome (DSS), 27 dengue fever (DF) and 63 age and sex matched controls, were included in this study. Blood samples were collected on the 3rd day of fever. Protein carbonylation (PCOs) and protein-bound sulphydryl (PBSH) group levels were determined by spectrophotometric method and analyzed as predictor of dengue hemorrhagic fever and dengue shock syndrome. RESULTS: About 80-84% of cases presented with no signs of DHF/DSS at the time of sampling. Dengue infected individuals had significantly elevated PCOs and low PBSH group levels than the controls. Using one-way ANOVA we found a significant difference with high PCOs and low PBSH group levels between DHF and DSS when compared with DF (P<0.001). However, no difference was observed in PBSH group levels between DHF and DSS. A significant difference in PCOs to PBSH ratio was observed among DF, DHF and DSS (P<0.001). Linear regression analysis revealed that duration of hospitalization is dependent on PCOs and PBSH group levels. Receiver operator curve (ROC) analysis indicated that 5.22nmol/mg protein PCOs; 1.08 PCOs to PBSH group levels ratio were optimal cutoff value for predicting DHF with sensitivity and specificity of 87.5% and 74.1%; 96.9% and 81.5%, respectively. For DSS prediction, 6.13 nmol/mg protein PCOs; 1.16 PCOs to PBSH group levels ratio were found as effective cutoff with sensitivity and specificity of 81% and 71.9%; 95.2% and 56.2%, respectively. CONCLUSION: Oxidative stress has been observed to develop since early days of onset of dengue infection. Plasma PCOs, PCOs to PBSH group ratio were found to very well predict DHF/DSS. PMID- 18420174 TI - Triaging patients with thyroid nodules for surgery by fine needle aspiration cytology. PMID- 18420175 TI - Effect of manual bowel decompression (milking) in the obstructed small bowel. AB - BACKGROUND: Mechanical intestinal obstruction is a frequently encountered problem in general surgery. One of the frequently used techniques for surgical decompression, so-called milking, is to caress the intestinal contents cephalad into the stomach or caudally into the colon. The aim of our study was to examine the functional, morphologic, and microbiologic effects of manual bowel decompression (milking) in the obstructed small bowel. METHODS: Six rats in the milking (M) group (obstructed and decompressed manually), 6 in the control (C) group (obstructed only), and 5 rats in the sham (S) group (laparotomy and evisceration) underwent surgery. Muscle contractility, gastrointestinal dye transmission, histopathologic changes of ileum, and bacterial translocation were analyzed. RESULTS: The contractile response of intestinal segments to acetylcholine was significantly less in group M compared with those of groups C and S (P < .05). The maximal contractile response to acetylcholine also was significantly lower in group M (P < .05). There was no statistically significant difference between the groups regarding the sensitivity of cholinergic receptors. Frequency of peristaltic movements, progression of Evans blue dye, histopathologic variables, and the quantitative evaluation (colony-forming units/gram of tissue) of isolated bacteria were not different among the groups. However, Escherichia coli in group M and Klebsiella spp in group S were the main isolated bacteria. CONCLUSIONS: Although it reduces muscle contractility, a milking procedure in an intestinal obstruction model does not cause peristaltic deterioration, histopathologic or inflammatory changes, or alterations in the degree of bacterial translocation. PMID- 18420177 TI - Cancer is not just a disease of a tissue: it is a host disease. How to reactivate host defense against tumors using conventional therapies of cancer? PMID- 18420176 TI - Laparoscopic ventral hernia repair under spinal anesthesia: a feasibility study. AB - BACKGROUND: Regional anesthesia has not been used as the sole anesthetic procedure in laparoscopic ventral hernia repair due to the fear of potential adverse effects of the pneumoperitoneum. However, there are recent reports on the feasibility of performing laparoscopic procedures, such as cholecystectomy, in fit patients, under spinal anesthesia alone. The current study aimed to detect the feasibility of performing laparoscopic ventral hernia repair under spinal anesthesia. METHODS: Twenty-five American Society of Anesthesiologists (ASA) I or II patients underwent laparoscopic ventral hernia repair with low-pressure CO2 pneumoperitoneum under spinal anesthesia. In 9 cases the hernia was umbilical/para-umbilical, in 5 cases epigastric, and in 11 cases incisional. Intraoperative incidents, complications, postoperative pain, and recovery in general, as well as patient satisfaction at follow-up examination, were prospectively recorded. RESULTS: All operations were completed laparoscopically and conversion from spinal to general anesthesia was not required in any of the cases. Median pain score at 4 hours postoperatively was .5 (range 0-5), at 8 hours 1.5 (range 0-6), and at 24 hours 1.5 (range 0-4). Most patients were discharged 24 hours after the operation; the median hospital stay was 1 day (range 1-3 days). At 2-weeks follow-up, no late complications were detected and all patients reported being satisfied with the anesthetic procedure. CONCLUSION: Laparoscopic ventral hernia repair with low-pressure CO2 pneumoperitoneum can be successfully and safely performed under spinal anesthesia. Furthermore, it seems that spinal anesthesia is associated with minimal postoperative pain and smooth recovery. PMID- 18420178 TI - New peptide-based therapies for metabolic diseases. PMID- 18420179 TI - [Stem cells in diabetes treatment]. PMID- 18420180 TI - Sensorimotor gating of schizophrenia patients is influenced by 5-HT2A receptor polymorphisms. AB - BACKGROUND: Schizophrenia patients exhibit impairment in prepulse inhibition (PPI) of the acoustic startle response (ASR), suggesting a sensorimotor gating deficit. The serotonin-2A receptor (5-HT(2A)R) has been implicated in both the pathogenesis of schizophrenia and the PPI deficits of schizophrenia patients. Moreover, both schizophrenia and PPI are thought to be inheritable. We investigated the impact of three 5-HT(2A)R polymorphisms (A-1438G, T102C, H452Y) on PPI in schizophrenia patients. METHODS: We analyzed the 5-HT(2A)R A-1438G, T102C, and H452Y polymorphisms and assessed startle reactivity, habituation, and PPI of ASR in 68 Caucasian schizophrenia inpatients. Patients were also examined with the Positive and Negative Syndrome Scale. RESULTS: The 5-HT(2A)R A-1438G and T102C polymorphisms were in complete linkage disequilibrium. Patients carrying the T102C TT and the A-1438G AA allele show significantly higher PPI levels and a faster early habituation compared with all other variants. 5-HT(2A)R A-1438G and T102C genotype explained approximately 11% of the PPI and early habituation variance. In contrast, the 5-HT(2A)R H452Y polymorphism did not affect startle parameters. CONCLUSIONS: Our findings suggest that PPI and habituation are modulated by 5-HT(2A)R A-1438G and T102C genotype in schizophrenia. Consequently, alterations within brain 5-HT(2A)Rs may contribute to the PPI deficits in schizophrenia. PMID- 18420181 TI - Effects of activated protein C on neonatal hypoxic ischemic brain injury. AB - Perinatal hypoxia-ischemia remains the single most important cause of brain injury in the newborn, leading to death or lifelong sequelae. White matter injuries in newborn infants have long-term effects on physical, visual, motor, sensory, cognitive and social development in human infants. There is no known cure for neonatal hypoxic ischemic encephalopathy (NHIE). Activated protein C has potent anticoagulant activity due to its ability to inactivate factor Va and VIIIa. APC is the first effective biological therapy approved for the treatment of severe sepsis. Although APC is well defined as a physiological anticoagulant, emerging data suggest that it also has cytoprotective, anti-inflammatory and antiapoptotic properties. APC has been shown to provide neuroprotection in ischemic brain and spinal cord injury. Here, we propose that APC, which modulates many of these processes, may represent a promising therapeutic agent for NHIE. Seven days old Wistar Albino rat pups have been used in the study (n=42). Experimental groups in the study were: sham-operated group, APC treated group, and vehicle treated group. In hypoxia-ischemia groups, the left common carotid artery was ligated permanently on the seventh postnatal day. Two hours after the procedure, hypoxia (92% nitrogen and 8% oxygen) was applied for 2.5 h. APC were injected (intraperitoneally; i.p.) as a single dose immediately after the hypoxia period. Brain nitrite levels, neuronal cell death, and apoptosis were evaluated in both hemispheres 72 h after the hypoxic-ischemic insult. Histopathological evaluation demonstrated that APC significantly diminished the number of "apoptotic cells" in the hippocampal CA1, CA2, CA3 and gyrus dentatus regions in both hemispheres. APC treatment significantly reduced "apoptotic cell death" in both hemispheres, when compared with vehicle treated group. APC significantly preserved the number of neurons CA1, CA3 regions of the hippocampus, when compared with vehicle treated group. Our results showed that hypoxic-ischemic injury caused a significant increase in NO production. The APC-treated animals were reduced brain nitrite levels in carotid ligated hemispheres. To our knowledge, this is the first study that demonstrates a protective effect of the APC against hypoxia-ischemia in the developing brain. PMID- 18420182 TI - Protein kinase C isozyme-specific potentiation of expressed Ca v 2.3 currents by acetyl-beta-methylcholine and phorbol-12-myristate, 13-acetate. AB - Protein kinase C (PKC) is implicated in the potentiation of Ca v 2.3 currents by acetyl-beta-methylcholine (MCh), a muscarinic M1 receptor agonist or phorbol-12 myristate, 13-acetate (PMA). The PKC isozymes responsible for the action of MCh and PMA were investigated using translocation as a measure of activation and with isozyme-selective antagonists and siRNA. Ca v channels were expressed with alpha1 2.3, beta1b and alpha2delta subunits and muscarinic M1 receptors in the Xenopus oocytes and the expressed currents (I Ba) were studied using Ba2+ as the charge carrier. Translocation of PKC isozymes to the membrane studied by Western blot revealed that all eleven known PKC isozymes are present in the Xenopus oocytes. Exposure of the oocytes to MCh led to the translocation of PKC alpha whereas PMA activated PKC betaII and epsilon isozymes. The action of MCh was inhibited by Go 6976, an inhibitor of cPKC isozymes or PKC alpha siRNA. PMA-induced potentiation of Ca v 2.3 currents was inhibited by CG533 53, a PKC betaII antagonist, betaIIV5.3, a peptide translocation inhibitor of PKC betaII or PKC betaII siRNA. Similarly, epsilonV1.2, a peptide translocation inhibitor of PKC epsilon or PKC epsilon siRNA inhibited PMA action. The inhibitors of PKC increased the basal I Ba slightly. It is possible that some PKC isozymes have negative control over the I Ba. Our results implicate PKC alpha in the potentiation of Ca v 2.3 currents by MCh and PKC betaII and epsilon in the potentiation of Ca v 2.3 currents by PMA. PMID- 18420183 TI - Effects of the task of categorizing FM direction on auditory evoked magnetic fields in the human auditory cortex. AB - We examined effects of the task of categorizing linear frequency-modulated (FM) sweeps into rising and falling on auditory evoked magnetic fields (AEFs) from the human auditory cortex, recorded by means of whole-head magnetoencephalography. AEFs in this task condition were compared with those in a passive condition where subjects had been asked to just passively listen to the same stimulus material. We found that the M100-peak latency was significantly shorter for the task condition than for the passive condition in the left but not in the right hemisphere. Furthermore, the M100-peak latency was significantly shorter in the right than in the left hemisphere for the passive and the task conditions. In contrast, the M100-peak amplitude did not differ significantly between conditions, nor between hemispheres. We also analyzed the activation strength derived from the integral of the absolute magnetic field over constant time windows between stimulus onset and 260 ms. We isolated an early, narrow time range between about 60 ms and 80 ms that showed larger values in the task condition, most prominently in the right hemisphere. These results add to other imaging and lesion studies which suggest a specific role of the right auditory cortex in identifying FM sweep direction and thus in categorizing FM sweeps into rising and falling. PMID- 18420184 TI - Curcumin protects against glutamate excitotoxicity in rat cerebral cortical neurons by increasing brain-derived neurotrophic factor level and activating TrkB. AB - Curcumin is a major active component isolated from Curcuma longa. Previously, we have reported its significant antidepressant effect. However, the mechanisms underlying the antidepressant effects are still obscure. In the present study, we explored the effect of curcumin against glutamate excitotoxicity, mainly focusing on the neuroprotective effects of curcumin on the expression of Brain-Derived Neurotrophic Factor (BDNF), which is deeply involved in the etiology and treatment of depression. Exposure of rat cortical neurons to 10 microM glutamate for 24 h caused a significant decrease in BDNF level, accompanied with reduced cell viability and enhanced cell apoptosis. Pretreatment of neurons with curcumin reversed the BDNF expression and cell viability in a dose- and time-dependent manner. However, K252a, a Trk receptor inhibitor which is known to inhibit the activity of BDNF, could block the survival-promoting effect of curcumin. In addition, the up-regulation of BDNF levels by curcumin was also suppressed by K252a. Taken together, these results suggest that the neuroprotective effect of curcumin might be mediated via BDNF/TrkB signaling pathway. PMID- 18420185 TI - Structural characterization of the acid-degraded secondary cell wall polymer of Geobacillus stearothermophilus PV72/p2. AB - The secondary cell wall polymer (SCWP) from Geobacillus stearothermophilus PV72/p2, which is involved in the anchoring of the surface-layer protein to the bacterial cell wall layer, is composed of 2-amino-2-deoxy- and 2-acetamido-2 deoxy-D-glucose, 2-acetamido-2-deoxy-D-mannose, and 2-acetamido-2-deoxy-D mannuronic acid. The primary structure of the acid-degraded polysaccharide- liberated by HF-treatment from the cell wall--was determined by high-field NMR spectroscopy and mass spectrometry using N-acetylated and hydrolyzed polysaccharide derivatives as well as Smith-degradation. The polysaccharide was shown to consist of a tetrasaccharide repeating unit containing a pyruvic acid acetal at a side-chain 2-acetamido-2-deoxy-alpha-D-mannopyranosyl residue. Substoichiometric substitutions of the repeating unit were observed concerning the degree of N-acetylation of glucosamine residues and the presence of side chain linked 2-acetamido-2-deoxy-beta-D-glucopyranosyl units: [Formula: see text]. PMID- 18420186 TI - Purification and characterization of a chitosanase from Serratia marcescens TKU011. AB - A chitosanase was purified from the culture supernatant of Serratia marcescens TKU011 with shrimp shell wastes as the sole carbon/nitrogen source. Zymogram analysis revealed the presence of chitosanolytic activity corresponding to one protein, which was purified by a combination of ion-exchange and gel-filtration chromatography. The molecular weight of the chitosanase was 21 kDa and 18 kDa estimated by SDS-PAGE and gel-filtration, respectively. The optimum pH, optimum temperature, pH stability, and thermal stability of the chitosanase were 5, 50 degrees C, pH 4-8, and <50 degrees C, respectively. The chitosanase was inhibited completely by EDTA, Mn(2+), and Fe(2+). The results of peptide mass mapping showed that three tryptic peptides of the chitosanase were identical to a chitin binding protein Cbp21 from S. marcescens (GenBank accession number gi58177632) with 63% sequence coverage. PMID- 18420187 TI - Protective effect of baicalin against lipopolysaccharide/D-galactosamine-induced liver injury in mice by up-regulation of heme oxygenase-1. AB - Baicalin, a traditional anti-inflammatory drug, has been found to protect against liver injury in several experimental animal hepatitis models; however, the mechanisms underlying the hepatoprotective properties of baicalin are poorly understood. In the present study,we investigated the effects of baicalin on the acute liver injury in mice induced by Lipopolysaccharide/D-galactosamine (LPS/D GalN). Baicalin (50, 150, and 300 mg/kg) was pretreated intraperitoneally (i.p.) at 2, 24, and 48 h respectively before LPS/D-GalN injected in mice. The mortality, hepatic tissue histology, hepatic tissue Tumor necrosis factor-alpha (TNF-alpha) and myeloperoxidase (MPO), plasma levels of TNF-alpha and alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were analyzed. Besides, western blotting analyses of nuclear factor kappa B (NF-kappaB) translocation and Heme oxygenase-1(HO-1) protein expression, as well as HO-1 activity were determined. The results showed that baicalin protected against LPS/D-GalN-induced liver injury, including dose-dependent alleviation of mortality and hepatic pathological damage, decrease of ALT/AST release and the rise of MPO. Baicalin reduced nuclear translocation of NF-kappa B, TNF-alpha mRNA and protein levels in hepatic tissues and plasma levels of TNF-alpha induced by LPS/D-GalN. Moreover, baicalin dose-dependently increased HO-1 protein expression and activity. Further, inhibition of HO-1 activity significantly reversed the protective effect of baicalin against LPS/D-GalN-induced liver injury. These results suggest that baicalin can effectively prevent LPS/D-GalN-induced liver injury by inhibition of NF-kappa B activity to reduce TNF-alpha production and the underlying mechanism may be related to up-regulation of HO-1 protein and activity. PMID- 18420188 TI - Protein kinase C is a common component of CGRP receptor desensitization induced by distinct agonists. AB - The calcitonin gene-related peptide (CGRP) is a neuropeptide involved in vasodilation and other physiological functions throughout the body. The receptor for CGRP has been cloned and well studied, but the mechanism of CGRP receptor desensitization has not been fully elucidated. In the present study, we evaluated the kinetics for agonist-mediated desensitization of the adenylate cyclase response in human neuroblastoma SK-N-MC cells. Distinct CGRP receptor agonists were used, including alpha and beta isoforms of CGRP, the linearized derivative cys(Et)2,7 alphaCGRP, adrenomedullin, and adrenomedullin 2. betaCGRP was 4-600 times more potent at desensitizing the cAMP production as compared to the other receptor-activating ligands, and all of the desensitization effects were blocked by a CGRP receptor antagonist. Although the different agonists vary in their ability to induce functional desensitization, a pretreatment/washout sequence with each peptide was able to reduce the activation potency of the other members of the calcitonin/CGRP peptide family. Next we tested whether the desensitizing effects of the distinct peptides involve protein kinase C (PKC) or protein kinase A (PKA). A PKC inhibitor, Ro 31-8220, concentration-dependently reduced the desensitization induced by the 5 CGRP receptor agonists, while having little effect on their desensitization potencies. PKA inhibitors KT-5720 and H-89, on the other hand, showed little effect on the induced level of desensitization. The findings indicate that functional desensitization is produced by distinct peptides acting through the active site of CGRP receptors, and involves the activation of PKC as a common component necessary to achieve maximal desensitization of receptor signaling. PMID- 18420189 TI - Mechanisms of the relaxant effect of vardenafil in rat penile arteries. AB - The aim of the present study was to investigate the mechanisms underlying the vasorelaxation induced by the selective phosphodiesterase 5 (PDE5) inhibitor vardenafil in rat penile small arteries. Segments of the rat dorsal penile artery were mounted in microvascular myographs for isometric tension recording. Concentration-response curves for vardenafil (1 nM-3 microM) and other PDE inhibitors (sildenafil, rolipram and milrinone) were constructed by adding cummulative concentrations of the drugs to arteries precontracted with phenylephrine. The effect of mechanical endothelial cell removal and of selective blockers of the nitric oxide (NO)/cGMP pathway and K+ channels were evaluated on the vardenafil relaxant responses. Vardenafil was the most potent of the four PDE inhibitors tested that maximally relaxed penile arteries, pD2 and maximum relaxation being 6.96+/-0.08 and 97+/-1% (n=48), respectively. Blockade of guanylate cyclase with ODQ (5 microM), mechanical removal of the endothelium or inhibition of NO synthase with l-NOARG (100 microM) markedly reduced vardenafil induced relaxations, without altering maximum response. Inhibitors of both the cGMP-dependent (PKG) and the cAMP-dependent (PKA) protein kinases, Rp-8-Br-PET cGMPS (5 microM) and Rp-8-CPT-cAMPS (50 microM), respectively, both reduced vardenafil relaxant responses and the later abolished that of rolipram. Vardenafil-elicited relaxation was reduced by the selective inhibitor of the large-conductance Ca2+-activated K+ channels (BK(Ca)), iberiotoxin (30 nM) and also by the ATP-sensitive K+ channel (K(ATP)) inhibitor, glibenclamide (1 microM). Vardenafil induces a potent vasodilatation in rat penile arteries that is partially dependent on the endothelium and the NO/cGMP pathway and involves activation of both BK(Ca) and K(ATP) channels. PMID- 18420190 TI - Behavioural genetics in mood and anxiety: a next step in finding novel pharmacological targets. AB - The pharmacological treatment of mood and anxiety disorders has for long relied on the serendipitous findings of monoaminergic and benzodiazepine drugs more than 50 years ago. These treatments, however, are therapeutically insufficient and even though more recently developed drugs are particularly improving side effects, the efficacy or response rate of the drugs has fundamentally not improved. Therefore it is necessary to develop new methods to identify novel mechanisms not based on merely the symptomatology, but on biologically relevant (endo) phenotypes. This review examines the option of integrating mouse and human behavioural genetics to aid the identification of the putative underlying pathophysiological mechanisms and pharmacological targets for psychiatric disorders. PMID- 18420191 TI - Translational aspects of pharmacological research into anxiety disorders: the stress-induced hyperthermia (SIH) paradigm. AB - In anxiety research, the search for models with sufficient clinical predictive validity to support the translation of animal studies on anxiolytic drugs to clinical research is often challenging. This review describes the stress-induced hyperthermia (SIH) paradigm, a model that studies the activation of the autonomic nervous system in response to stress by measuring body temperature. The reproducible and robust SIH response, combined with ease of testing, make the SIH paradigm very suitable for drug screening. We will review the current knowledge on the neurobiology of the SIH response, discuss the role of GABA(A) and serotonin (5-HT) pharmacology, as well as how the SIH response relates to infectious fever. Furthermore, we will present novel data on the SIH response variance across different mice and their sensitivity to anxiolytic drugs. The SIH response is an autonomic stress response that can be successfully studied at the level of its physiology, pharmacology, neurobiology and genetics and possesses excellent animal-to-human translational properties. PMID- 18420192 TI - Sphingolipid signalling in the cardiovascular system: good, bad or both? AB - Sphingolipids are biologically active lipids that play important roles in various cellular processes and the sphingomyelin metabolites ceramide, sphingosine and sphingosine-1-phosphate can act as signalling molecules in most cell types. With the recent development of the immunosuppressant drug FTY720 (Fingolimod) which after phosphorylation in vivo acts as a sphingosine-1-phosphate receptor agonist, research on the role of sphingolipids in the immune and other organ systems was triggered enormously. Since it was reported that FTY720 induced a modest, but significant transient decrease in heart rate in animals and humans, the question was raised which pharmacological properties of drugs targeting sphingolipid signalling will affect cardiovascular function in vivo. The answer to this question will most likely also indicate what type of drug could be used to treat cardiovascular disease. The latter is becoming increasingly important because of the increasing population carrying characteristics of the metabolic syndrome. This syndrome is, amongst others, characterized by obesity, hypertension, atherosclerosis and diabetes. As such, individuals with this syndrome are at increased risk of heart disease. Now numerous studies have investigated sphingolipid effects in the cardiovascular system, can we speculate whether certain sphingolipids under specific conditions are good, bad or maybe both? In this review we will give a brief overview of the pathophysiological role of sphingolipids in cardiovascular disease. In addition, we will try to answer how drugs that target sphingolipid signalling will potentially influence cardiovascular function and whether these drugs would be useful to treat cardiovascular disease. PMID- 18420193 TI - The beta-chemokines CCL2 and CCL7 are two novel differentiation factors for midbrain dopaminergic precursors and neurons. AB - beta-chemokines are secreted factors that regulate diverse functions in the adult brain, such as neuro-immune responses and neurotransmission, but their function in the developing brain is largely unknown. We recently found that the orphan nuclear receptor, Nurr1, up regulates CCL2 and CCL7 in neural stem cells, suggesting a possible function of beta-chemokines in midbrain development. Here we report that two beta-chemokines, CCL2 and CCL7, and two of their receptors, CCR1 and CCR2, are expressed and developmentally regulated in the ventral midbrain (VM). Moreover, we found that the expression of CCL7 was down regulated in the Nurr1 knockout mice, linking CCL7 to dopamine (DA) neuron development. When the function of CCL2 and CCL7 was examined, we found that they selectively enhanced the differentiation of Nurr1+ precursors into DA neurons, but not their survival or progenitor proliferation in primary precursor cultures. Moreover, both CCL2 and CCL7 promoted neuritogenesis in midbrain DA neuron cultures. Thus, our results show for the first time a function of beta-chemokines in the developing brain and identify beta-chemokines as novel class of pro differentiation factors for midbrain DA neurons. These data also suggest that beta-chemokines may become useful tools to enhance the differentiation of DA cell preparations for cell replacement therapy and drug discovery in Parkinson's disease (PD). PMID- 18420194 TI - The specific contribution of hypoxia-inducible factor-2alpha to hypoxic gene expression in vitro is limited and modulated by cell type-specific and exogenous factors. AB - Cellular integrity in hypoxia is dependent on molecular adaptations dominated by the heterodimeric transcription factor hypoxia-inducible factor (HIF). The HIF complex contains one of two alternative oxygen-regulated alpha-subunits considered to play distinct roles in the hypoxia response. Although HIF-2alpha may be more important in tumour biology and erythropoiesis, the spectrum of individual target genes is still insufficiently characterized. We therefore performed an Affymetrix gene array on Hep3B cells stimulated with a hypoxia mimetic and transfected with either HIF-1alpha or HIF-2alpha siRNA. 271 transcripts were found to be induced HIF-dependently, including most previously identified HIF targets and a number of novel genes. Most were influenced by HIF 1alpha knock-down, whereas a smaller number were regulated by HIF-2alpha. Validation of a selection of genes by RNase protection confirmed the hypoxic regulation and HIF-1alpha- or HIF-2alpha-dependency in most cases, with the latter showing a lower amplitude. Many HIF-2alpha targets also responded to HIF 1alpha knock-down. Interestingly, regulation by HIF-2alpha was markedly influenced not only by cell type, but also by cell culture conditions, features that were not shared with HIF-1alpha-regulated genes. Thus, HIF-2alpha effects are modulated by a number of intrinsic and extrinsic factors which may be most relevant in tumour cells. PMID- 18420195 TI - In vitro benzyl alcohol cytotoxicity: implications for intravitreal use of triamcinolone acetonide. AB - The aim of the study was to investigate the toxicity of benzyl alcohol (BA), the preservative in commercial triamcinolone acetonide (TA) suspensions, on retinal pigment epithelial (RPE) cells. Cultured RPE cells from a human cell line (ARPE 19) and from rabbits were exposed to the balanced salt solution (control) or BA (0.0225, 0.225, 0.9, 3 or 9mg/mL) for 5, 30, 60, or 120min. Morphological changes of RPE cells were evaluated by the trypan blue in situ staining. The proportions of dead cells were quantitatively measured by the trypan blue exclusion assay, and those of functional cells were assessed by a mitochondrial dehydrogenase assay. The mechanism of cytotoxicity was determined by the acridine orange/ethidium bromide staining and DNA laddering technique. Furthermore, ultrastructural changes were observed by transmission electron microscopy. The results showed that RPE cell damage was dose- and time-dependent. BA 0.225mg/mL, the clinically relevant concentration in TA following intravitreal injection, caused ultrastructural damage and impaired human RPE cell function at 2h; but BA 0.0225mg/mL did not. BA 9.0mg/mL, the concentration in commercial TA suspensions, was toxic within 5min on each assay for both human and rabbit RPE cells. The major mechanism of cell death was necrosis. In conclusion, BA in commercial TA suspensions injected intravitreally (0.225-9mg/mL) can damage RPE cells. Our in vitro study on benzyl alcohol cytotoxicity has significant clinical implications for intravitreal use of TA. We suggest that, before a commercial TA solution is used intravitreally, the vehicle should be removed to prevent damaging the RPE layer, particularly during macular hole surgery. Commercial development of a preservative-free TA suspension for intraocular use is urged. PMID- 18420196 TI - Retinal and choroidal vasoreactivity to altered PaCO2 in rat measured with a modified microsphere technique. AB - Rats are increasingly used in ophthalmic research. However, little is known about the metabolic regulation of ocular blood flow. The purpose of this study was to examine the vasoreactivity in retina and choroid of the rat eye in response to experimentally altered partial arterial pressure of CO2 (PaCO2). The retinal and choroidal blood flows were measured sequentially in different PaCO2 with a modified microspheres method. The experiments were performed in two groups of adult male Brown-Norway rats. Under isofluorane anesthesia and mechanical ventilation, PaCO2 was monitored continuously by recording end tidal carbon dioxide level. Both femoral arteries and a femoral vein were cannulated for arterial blood pressure monitoring, blood sampling and drug administration, respectively. The intraocular pressure in both eyes was manometrically controlled at 20mmHg by anterior chamber cannulation. The retinal and choroidal blood flows were simultaneously measured by cardiac injection of a mixture containing 3.75 million of 8microm, and 0.5 million of 10microm microspheres; each size having a distinct color. In one experiment (n=10), blood flow was first measured during normocapnia (PaCO2=35mmHg) and then during hypocapnia (PaCO2=20-25mmHg). In another experiment (n=7), blood flow was measured during hypercapnia (PaCO2=45 50mmHg) and repeated one more time under the same experimental conditions to evaluate the repeatability of sequential measurements and the variances of the measurement between the two eyes. The results show that the mean blood flow in the retina measured during hypocapnia, normocapnia and hypercapnia were 8.1+/ 4.8, 15.1+/-8.5 and 27.4+/-4.6microl/min per tissue, respectively. In the choroid, the corresponding blood flow rates were 120+/-38, 166+/-28 and 149+/ 28microl/min per tissue, respectively. The difference of the mean blood flows across all the three different PaCO2 groups was highly significant for both retina and choroid (ANOVA: P<0.0001 and P=0.01, respectively). The mean blood flow during hypocapnia was significantly lower than normocapnia in both retina and choroid (P<0.02). The blood flow under hypercapnia was significantly higher than normocapnia in retina (P<0.01), but not in choroid (P=0.62). In conclusion, the study demonstrated that the dual-size and dual-dose microspheres mixture can be used as a reliable method to measure the retinal and choroidal blood flows simultaneously and sequentially in rats. The vasoreactivity to altered systemic PaCO2 in the retina in rats is similar to that of most other species studied. However, the choroidal vascular system exhibited complicated features that remain to be further clarified. PMID- 18420198 TI - Early-life status epilepticus induces ectopic granule cells in adult mice dentate gyrus. AB - A large number of aberrant hilar granule cells (GCs) are found in the patients and animal models of adult temporal lobe epilepsy (TLE), and these "ectopic" GCs have synchronous epileptiform bursting with other hippocampal neurons. In this study, we investigated whether early-life status epilepticus (SE) induces hilar ectopic GCs that remain in the adulthood because TLE patients frequently experience seizures in the early childhood when a large number of postnatally born GCs migrate in the hilus. To label newborn GCs, bromodeoxyuridine (BrdU) was injected daily for three consecutive days to C57BL/6J mice at different postnatal days starting at postnatal-0-day-old (P0) (Group1), P7 (Group2), or P35 (Group3). Mice in each group underwent pilocarpine-induced SE at P14. Six months later, to determine whether SE induces ectopic GCs, we plotted the distribution of postnatally born GCs which were immunohistochemically defined as BrdU- and the GC marker Prox1-colabeled cells. We also examined whether SE causes the granule cell layer (GCL) dispersion and/or the mossy fiber (MF) sprouting, other representative pathologies of TLE hippocampus. Only SE-experiencing mice in Group1 had significantly more neonatally born ectopic GCs compared with control mice. Neither control nor SE mice had dispersed GCL. All mice that underwent SE had sprouted MFs in CA3. We conclude that early-life SE disrupts a normal incorporation of GCs born pre-SE but not post-SE, inducing ectopic GCs in the adult hilus. Interestingly, the results also indicate that developmentally earlier born GCs are more responsive to early-life SE in terms of the emergence of ectopic GCs. PMID- 18420197 TI - Endothelin-1 mediated regulation of extracellular matrix collagens in cells of human lamina cribrosa. AB - Endothelin-1 (ET-1), a potent vaso-active peptide, mediates extracellular matrix regulation resulting in an increase in collagen deposition in various cell types and tissues and has been proposed to play a key role in glaucoma pathology. The role of ET-1 in the regulation of extracellular matrix collagens at the level of optic nerve head is not known. In this study we have examined the role of ET-1 in extracellular matrix collagen regulation in primary cultures of human lamina cribrosa cells. Our hypothesis is that ET-1 increases remodeling of the ECM of cells of the lamina cribrosa. Such actions could contribute to the development of optic neuropathy. QPCR analysis revealed that ET-1 mediated an increase in mRNA levels of collagen type I alpha1 and collagen type VI alpha1 chains at all doses of ET-1 with a significant increase at 1nM and 10nM concentration in LC cells. A dose-dependent increase in collagen type I and type VI protein deposition and secretion was also observed by Western blot in response to ET-1 and was significant at 10nM and 100nM concentrations of ET-1. ET-1 increased the [3H] proline uptake in LC cells suggesting that ET-1 contributed to an increase in total collagen synthesis in LC cells. ET-1-mediated increase in collagen type I, type VI and total collagen synthesis was significantly blocked by the ET(A) receptor antagonist, BQ610, as well as with the ET(B) receptor antagonist, BQ788, suggesting the involvement of both receptor subtypes in ET-1 mediated collagen synthesis in LC cells. These results suggest that ET-1 regulates extracellular matrix collagen synthesis in LC cells and may contribute to ECM remodeling at the level of LC of POAG subjects who have elevated plasma and aqueous humor levels of endothelin-1. PMID- 18420199 TI - Treadmill training promotes axon regeneration in injured peripheral nerves. AB - Physical activity after spinal cord injury promotes improvements in motor function, but its effects following peripheral nerve injury are less clear. Although axons in peripheral nerves are known to regenerate better than those in the CNS, methods of accelerating regeneration are needed due to the slow overall rate of growth. Therefore we studied the effect of two weeks of treadmill locomotion on the growth of regenerating axons in peripheral nerves following injury. The common fibular nerves of thy-1-YFP-H mice, in which a subset of axons in peripheral nerves express yellow fluorescent protein (YFP), were cut and repaired with allografts from non-fluorescent littermates, and then harvested two weeks later. Mice were divided into groups of low-intensity continuous training (CT, 60 min), low-intensity interval training (IT; one group, 10 reps, 20 min total), and high-intensity IT (three groups, 2, 4, and 10 reps). One repetition consisted of 2 min of running and 5 min of rest. Sixty minutes of CT resulted in the highest exercise volume, whereas 2 reps of IT resulted in the lowest volume of exercise. The lengths of regenerating YFP(+) axons were measured in images of longitudinal optical sections of nerves. Axon profiles were significantly longer than control in all exercise groups except the low-intensity IT group. In the CT group and the high-intensity IT groups that trained with 4 or 10 repetitions axons were more than twice as long as unexercised controls. The number of intervals did not impact axon elongation. Axon sprouting was enhanced in IT groups but not the CT group. Thus exercise, even in very small quantities, increases axon elongation in injured peripheral nerves whereas continuous exercise resulting in higher volume (total steps) may have no net impact on axon sprouting. PMID- 18420200 TI - State-dependent slow outlasting activities following neocortical kindling in cats. AB - Some forms of electrographic seizures are generated at the level of the cortical network. Neocortical kindling exhibits a resistance to produce generalized convulsive seizures, and therefore, it was rather difficult to use it to study the cortical epileptogenesis. Here, using supra-threshold cortical kindling, we report electrophysiological patterns of field-potential synchronization and intracellular activities in chronically implanted non-anesthetized cats, during different states of vigilance, and during acute seizures elicited by prolonged (20-60 s) electrical stimulation. Acute seizures were easily elicited during transition from slow-wave sleep (SWS) to waking state. The seizures were mainly clonic accompanied with tonic components followed by prolonged postictal depression. Delayed rhythmic outlasting activities (OA) at approximately 1.5 Hz, first time reported here, followed the postictal depression, and lasted up to 2 h. These activities were clear during waking state, slightly reduced during SWS and completely absent during rapid-eye movement sleep. They started focally and following daily stimulations generalized over the entire cortical surface. Extra- and intracellular neuronal recordings during OA displayed spike-doublets, built on the summation of successive excitatory postsynaptic potentials and fast prepotentials, entailing an increased dendritic excitation. Our results suggest that such rhythmic long-lasting oscillatory activity outlasting seizures are the key factor of epileptogenesis, leading to epilepsy. PMID- 18420201 TI - Variation in somatic and ovarian development: predicting susceptibility of amphibians to estrogenic contaminants. AB - Although amphibian sex determination is genetic, it can be manipulated by exogenous hormone exposure during sexual differentiation. The timing of sexual differentiation varies among anuran amphibians such that species may or may not be a tadpole during this period, and therefore, may or may not be exposed to aquatic contaminants. Estrogenic contamination is present in amphibian habitats worldwide. We examined three species with varying somatic and ovarian developmental rates to assess their susceptibility to estrogenic contaminants. American toads (Bufo americanus), gray treefrogs (Hyla versicolor), and Southern leopard frogs (Ranasphenocephala) were exposed as larvae to 17-beta-estradiol (10(-7)M), three concentrations of a widespread herbicide (1, 3, 30 ppb atrazine), or a solvent control (ethanol). Somatic and ovarian developmental stages as well as time to metamorphosis were recorded. Toads and treefrogs were examined at three weeks and metamorphosis, while leopard frogs were examined at three, six, and nine weeks as well as at metamorphosis. Our results demonstrate that each species displays heterochronic somatic and ovarian development. Further, the more rapid of the two rates determines the susceptibility to estrogenic contaminants. These results suggests that amphibians with shorter larval periods, and therefore quicker somatic developmental rates (i.e. American toads, gray treefrogs), are more susceptible to somatic treatment effects (i.e. prolonged time to metamorphosis) due to estrogenic contaminants. Moreover, the results suggest that amphibians with relatively rapid ovarian development (i.e. Southern leopard frogs) are more susceptible to gonadal treatment effects caused by estrogenic contaminants. PMID- 18420202 TI - Two-stage robust control of substrate concentration for an activated sludge process. AB - A two-stage robust control scheme improving the performance of an Activated Sludge Process is proposed. In the first stage, asymptotic command following the substrate concentration with simultaneous attenuation of the fluctuations of the dissolved oxygen concentration is assured. The first stage is a pure dynamic controller. The second stage is a PID controller. Good performance of the proposed control scheme on the corresponding nonlinear ASP model is illustrated through extensive simulation experiments. The contribution of the paper can be summarized to the derivation of the following two results: An accurate to a wide range of inputs and disturbances, linearized generic model of the ASP and, most important, a linear robust controller that controls accurately the effluent substrate concentration without using measurements of it. PMID- 18420203 TI - Putative proprioceptive function of the pelvic ligaments: biomechanical and histological studies. AB - The sacrospinous (SS) and sacrotuberous (ST) ligaments of the pelvic ring are known as mechanical stabilisers of the pelvic girdle, primarily against rotational forces in the sagittal and horizontal planes. Earlier studies, however, raised the possibility that ST/SS ligaments possess significant proprioceptive function, while the mechanical role of these ligaments in maintaining the structural integrity of the pelvis is of less importance. The aim of this study is to determine whether the function of these ligaments is strictly to provide mechanical stability or if they have any additional functional properties, i.e., proprioception. In order to reveal the function of the SS/ST ligaments, biomechanical studies of cadaver pelvis were used along with the histological analysis of the ligaments. Following measurements to determine the accurate mechanical role of the pelvic ligaments, the strength of these ligaments was significantly less than we earlier expected. For this reason other functions of the SS/ST ligaments were considered, including the proprioceptive role. Indeed, histological studies revealed ramifying nerve terminals in the SS/ST ligaments. These terminals may represent the morphological substrate of the proprioceptive function associated with the ligaments. Our studies revealed that SS/ST ligaments might have a significant proprioceptive function providing information of the position of the pelvis. Consequently, the mechanical role of the ligaments in maintaining the structural integrity of the pelvis may be significantly less than previously assumed. Understanding the function of the SS/ST ligaments is crucial for providing more precise guidelines for patient management with injuries to the posterior pelvic region. PMID- 18420204 TI - Trends in major modifiable risk factors for severe perineal trauma, 1996-2006. AB - OBJECTIVE: To examine what effect the major modifiable risk factors for severe perineal trauma have had on the rates of this trauma over time. METHODS: A retrospective observational cohort study of singleton vaginal deliveries taken from a perinatal database for the period 1996 through 2006. RESULTS: A total of 46,239 singleton vertex vaginal deliveries met the inclusion criteria. Major risk factors for severe perineal trauma were increased maternal age (odds ratio [OR] 1.28, 95% confidence interval [CI] 1.1-1.5), non-African American ethnicity (OR 1.5, 95% CI 1.3-1.7), nulliparity (OR 4.8, 95% CI 4.11-5.6), fetal birth weight (OR 2.2, 95% CI 1.9-2.4), forceps (OR 8.3, 95% CI 5.4-10.8), vacuum (OR 2.9, 95% CI 1.9-4.4), and midline episiotomy (OR 5.7, 95% CI 5.0-6.4). Evaluation of the changes in rates of these factors over the study period revealed that the decline in the rates of episiotomy and the use of forceps accounted for a reduction in severe lacerations of more than 50%. CONCLUSION: Reduction of severe perineal trauma by restricted use of the 2 modifiable clinical variables, episiotomy and forceps, is evident over time. PMID- 18420205 TI - Neuropeptide Y expression in vaginal epithelium of women with pelvic organ prolapse and stress urinary incontinence. AB - OBJECTIVES: To determine the role of neuropeptide Y (NPY) in anterior and posterior vaginal epithelium in the etiologic development of pelvic organ prolapse (POP) and stress urinary incontinence (SUI). METHODS: Forty biopsy specimens from anterior and posterior vaginal epithelium were obtained from 40 POP/SUI patients and controls. The specimens were stained using hematoxylin and eosin and NPY immunohistochemical staining. NPY was measured semiquantitatively and NPY mRNA expression was assessed using DNA hybridization in situ. RESULTS: There were no significant differences in NPY between anterior and posterior vaginal epithelium. NPY profiles in posterior vaginal epithelium in the SUI group were significantly lower than in the POP (P<0.05) and control (P<0.05) groups. In the POP group, the NPY profile correlated negatively with advancing age and years post menopause. CONCLUSION: The reduction in NPY in the anterior and posterior vaginal wall epithelium might be related to nerve damage or degeneration, resulting in a change in blood flow, atrophy, and pelvic floor laxity in patients with POP and SUI, especially post menopause and with advancing age. PMID- 18420206 TI - Unequal tissue expression of proteins from the PA/PAI system, myoma necrosis, and uterus survival after uterine artery occlusion. AB - OBJECTIVE: To further the understanding of the mechanism behind myoma necrosis and uterus survival after uterine artery occlusion. METHOD: Myomal and myometrial gene transcription and protein expression of tissue plasminogen activator (tPA), urokinase plasminogen activator (uPA), and plasminogen activator inhibitor type 1 (PAI-1) were investigated in 30 uteri. RESULTS: Compared with myometrial expression, myomal expression was less for uPA mRNA (1.34%+/-2.18% vs 2.84%+/ 2.97%; P<0.05) and greater for PAI-1 mRNA (2.67%+/-1.82% vs 1.27%+/-1.99%; P<0.05), but the difference in tPA mRNA expression was not significant. Accordingly, compared with myometrial expression, myomal expression was less for the uPA protein (P<0.05) and greater for the PAI-1 protein (P<0.05), but the difference in tPA protein was not significant. CONCLUSION: The PA/PAI system was found to be expressed differently in myomas and the myometrium, which may play a role in uterus survival and myoma necrosis after uterine artery occlusion. PMID- 18420207 TI - Sexual function in women following the transvaginal tension-free tape procedure for incontinence. AB - OBJECTIVE: To assess the impact of the tension-free vaginal tape (TVT) procedure on the sexual function of women who underwent this treatment for stress urinary incontinence (SUI). METHOD: The Lemack questionnaire was mailed to 135 women treated for SUI only, with no concomitant pelvic organ prolapse repair. RESULTS: Of the 82 returned questionnaires (60.7%), 66 (80.5%) could be included in the analysis. Compared with preoperative responses, there were no significant postoperative changes regarding frequency of sexual intercourse, satisfaction with sexual intercourse, or personal importance of having an active sexual life. Although there was a significant postsurgical decrease in urinary coital incontinence (P=0.02) and 12 women (25.5%) reported improved satisfaction from intercourse, 11 women (23.4%) complained of a worsening. CONCLUSION: These results suggest that TVT for SUI does not significantly affect sexual function in women. Additional prospective studies are warranted to verify these preliminary findings and compare the impact of the TVT with that of other anti-incontinence procedures. PMID- 18420208 TI - Low back pain characteristics from undergraduate student to working nurse in Australia: a cross-sectional survey. AB - BACKGROUND: Nurses are known to be a high risk group for occupational low back pain (LBP). The periods of greatest risk for developing low back pain in this population are not well defined. Recent literature suggests current preventative strategies are not consistently effective in improving low back injury statistics among health care populations. OBJECTIVES: To identify the relative contributions of age and occupational exposure on the prevalence, duration and severity of low back pain episodes among undergraduate nursing students and recently graduated nurses. DESIGN: Cross-sectional survey. SETTINGS: Two university undergraduate nursing schools and one public teaching hospital graduate nurse training program in Western Australia. PARTICIPANTS: 897 undergraduate nursing students (years 1, 2 and 3) and 111 graduate nurses recruited by personal invitation during lectures. METHODS: Using a modified version of the Nordic Low Back Questionnaire, information regarding low back pain episode prevalence, impact, duration, frequency and causes was obtained. RESULTS: Mean age was consistent across all groups (26.7+/-9.0 years) and had no significant effect on lifetime low back pain prevalence (p=0.30). Very high lifetime (79%), 12 month (71%) and 7 day (31%) low back pain prevalence rates were consistent across all 3 year groups of undergraduate nursing students, but were significantly higher after 12 months of full-time employment [lifetime (95.5%), 12 month (90%) and 7 day (39%)]. Around 60% of all respondents with low back pain utilised at least one of (a) treatment, (b) medication, or (c) a reduction in activity. Nursing students and graduate nurses attributed the majority of their low back pain to bending or lifting despite recent efforts to reduce manual workplace demands (lifting) on nurses. Strategies for managing low back pain differed between nursing students and graduate nurses. CONCLUSIONS: These results may suggest a rise in occupational exposure from student to working nurse is the primary cause of the increase in low back pain. Increased exposure may be to physical as well as psychological stressors. Given that prevalence rates are very high prior to commencing work, nursing student populations should be a target group for low back pain preventative strategies. PMID- 18420209 TI - ICAM1 and VCAM1 polymorphisms, coronary artery calcium, and circulating levels of soluble ICAM-1: the multi-ethnic study of atherosclerosis (MESA). AB - Intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) may be important contributors to the development and progression of atherosclerosis. Using a stratified random sample of 2880 participants of the Multi-Ethnic Study of Atherosclerosis we investigated the relationship of 12 ICAM1 and 17 VCAM1 SNPs and coronary artery calcium (CAC) and ICAM1 SNPs and circulating levels of soluble ICAM-1 (sICAM-1). There were no ICAM1 or VCAM1 SNPs significantly associated with CAC in any of the four race/ethnic groups. In a subset of 1451 subjects with sICAM-1 measurements, we observed a significant association with rs5491 in all four race/ethnic groups corroborating previous research that has shown that the T-allele of rs5491 interferes with the monoclonal antibody used to measure sICAM-1 in this study. After excluding all rs5491 T-allele carriers, several ICAM1 SNPs were significantly associated with sICAM-1 levels; rs5496 in African Americans, rs5498 and rs3093030 in European Americans, and rs1799969 in Hispanics. Our results identified ICAM1 polymorphisms that were significantly associated with sICAM-1 level but not CAC, a subclinical marker of atherosclerosis. PMID- 18420210 TI - High-density lipoprotein subclasses distribution and composition in Mexican adolescents with low HDL cholesterol and/or high triglyceride concentrations, and its association with insulin and C-reactive protein. AB - We tested whether low high-density lipoprotein cholesterol (HDL-C) and/or high triglycerides are associated to abnormal HDL subclasses distribution and composition, and their relationships with fasting insulin and C-reactive protein (CRP). Four groups of adolescents were studied: group 1 (HDL-C< or =35 mg/dl+TG> or =150 mg/dl; n=16); group 2 (isolated HDL-C< or =35 mg/dl; n=31); group 3 (isolated TG> or =150 mg/dl; n=20); and group 4 (CT<200 mg/dl, HDL-C>35 mg/dl, LDL-C<130 mg/dl, and TG<150 mg/dl; n=39). Tanner score-adjusted proportions of large subspecies (HDL(2b), HDL(2a)) were lower, and small (HDL(3b), HDL(3c)) were higher in groups 1, 2 and 3 than in group 4. As a result, HDL particle size in the three dyslipidemic groups was smaller than in group 4 (p<0.001). HDL CE, FC, PL, and apo AI percent contents were lower, whereas HDL TG percent content was higher in groups 1, 2 and 3 compared to group 4. CRP median values were also significantly higher in the three groups with dyslipidemia than in normolipidemic subjects (group 4). Fasting Insulin concentration and HOMA-IR were significantly higher in group 1 than in the other three groups. In stepwise multivariate analysis HDL subclass distribution and composition were independently associated only with HDL-C and waist circumference. As reported in adults, adolescents with low HDL-C and/or high TG have abnormalities in HDL subclasses distribution and lipid composition, which may render their HDL dysfunctional. In addition, these subjects have high CRP and insulin levels suggesting the presence of chronic low grade inflammation. PMID- 18420211 TI - Do we need to consider inflammatory markers when we treat atherosclerotic disease? AB - This review considers the value of monitoring inflammatory markers as a guide to selecting appropriate drugs in patients at high risk of cardiovascular disease (CVD). Clinical and experimental studies investigated inflammation in patients with acute coronary syndromes (ACS), stable coronary artery disease (CAD) and diabetes mellitus (DM) or metabolic syndrome (MetS), non-alcoholic fatty liver disease (NAFLD) or systemic autoimmune diseases (SAD). Evidence suggests that in these high risk groups inflammation plays a role in the extent and severity of atherosclerosis. Simple inflammatory markers (e.g. C-reactive protein and fibrinogen) can be monitored cost effectively and may influence the selection of drugs that can normalize both traditional CVD risk factors and inflammation. However, this concept requires proof in appropriately designed trials that include clinically relevant end points. PMID- 18420212 TI - Thioredoxin in coronary culprit lesions: possible relationship to oxidative stress and intraplaque hemorrhage. AB - The present study investigated the expression of thioredoxin (TRX), an important anti-oxidative protein, and its relationship to plaque instability in atherectomy specimens from 43 and 42 patients with stable (SAP) and unstable (UAP) angina pectoris, respectively. We histologically assessed thrombus formation, cellular elements, localization of TRX and of oxidized low density lipoprotein (ox-LDL), intraplaque hemorrhage, and transition metal iron (Fe(2+), Fe(3+)) deposition in these specimens. The clinical characteristics of the two groups did not differ except for aspirin administration. The incidence of thrombus formation was more frequent (P=0.005) and immunopositive areas of macrophage, TRX and ox-LDL were significantly larger in patients with UAP than SAP (P<0.001, each). Macrophages were mainly immunoreactive for TRX and ox-LDL. Intraplaque hemorrhage evaluated by glycophorin A immunoreactivity and Fe(2+)/Fe(3+) deposition was also more obvious in lesions from patients with UAP than SAP (P<0.001, each). Additionally, immunopositive areas of TRX and ox-LDL positively correlated with Fe(2+)/Fe(3+) deposition and were also associated with thrombus formation. Although the underlying mechanisms remain unknown, TRX was up-regulated in response to increased oxidative stress and associated with intraplaque hemorrhage of coronary culprit lesions, and thus might be a potent marker of plaque instability. PMID- 18420213 TI - A musculoskeletal model of the upper extremity for use in the development of neuroprosthetic systems. AB - Upper extremity neuroprostheses use functional electrical stimulation (FES) to restore arm motor function to individuals with cervical level spinal cord injury. For the design and testing of these systems, a biomechanical model of the shoulder and elbow has been developed, to be used as a substitute for the human arm. It can be used to design and evaluate specific implementations of FES systems, as well as FES controllers. The model can be customized to simulate a variety of pathological conditions. For example, by adjusting the maximum force the muscles can produce, the model can be used to simulate an individual with tetraplegia and to explore the effects of FES of different muscle sets. The model comprises six bones, five joints, nine degrees of freedom, and 29 shoulder and arm muscles. It was developed using commercial, graphics-based modeling and simulation packages that are easily accessible to other researchers and can be readily interfaced to other analysis packages. It can be used for both forward dynamic (inputs: muscle activation and external load; outputs: motions) and inverse-dynamic (inputs: motions and external load; outputs: muscle activation) simulations. Our model was verified by comparing the model calculated muscle activations to electromyographic signals recorded from shoulder and arm muscles of five subjects. As an example of its application to neuroprosthesis design, the model was used to demonstrate the importance of rotator cuff muscle stimulation when aiming to restore humeral elevation. It is concluded that this model is a useful tool in the development and implementation of upper extremity neuroprosthetic systems. PMID- 18420214 TI - Active and passive contributions to joint kinetics during walking in older adults. AB - The objectives of this study were to characterize the active and passive contributions to joint kinetics during walking in healthy young and older adults, and assess whether isokinetic ankle strength is associated with ankle power output during walking. Twenty healthy young (18-35 years) and 20 healthy older (65-85 years) adults participated in this study. We measured subject-specific passive-elastic joint moment-angle relationships in the lower extremity and tested maximum isokinetic ankle strength at 30 deg/s. Passive moment-angle relationships were used to estimate active and passive joint moment, power, and work quantities during walking at 80%, 100% and 120% of preferred walking speed. There were no significant differences in walking speed, step length, or cadence between the older and young adults. However, the older adults produced significantly more net positive work at the hip but less net positive work at the ankle at all walking speeds. Passive contributions to hip and ankle work did not significantly differ between groups, inferring that the older adults generated the additional hip work actively. Maximum isokinetic ankle strength was significantly less in the older adults, and correlated with peak positive plantar flexor power at both the preferred and fast walking speeds. The results of this study suggest that age-related shifts in joint kinetics do not arise as a result of increased passive hip joint stiffness, but seem to be reflected in plantar flexor weakness. PMID- 18420215 TI - Sol-gel methyl coating in capillary microextraction hyphenated on-line with high performance liquid chromatography Counterintuitive extraction behavior for polar analytes. AB - A sol-gel coating with anchored methyl groups was developed for capillary microextraction hyphenated on-line with high-performance liquid chromatography (HPLC). This was accomplished by using methyltrimethoxysilane as the sol-gel precursor. The methyl group on the sol-gel precursor ultimately turned into a pendant group on the created sol-gel coating and was primarily responsible for the extraction of nonpolar analytes. A 40-cm segment of 0.25mm I.D. fused silica capillary containing the sol-gel methyl coating on the inner surface was installed as a sampling loop in an HPLC injection port. The analytes were extracted by the coating when an aqueous sample containing the analytes was passed through this capillary. The extracted analytes were then transferred to the HPLC column using isocratic elution with an acetonitrile/water mobile phase. This capillary demonstrated excellent extraction capability for polycyclic aromatic hydrocarbons and ketones. Unexpectedly, this coating also provided good extraction for polar analytes, including aromatic phenols, alcohols, and amines. Considering the fact that the methyl group is nonpolar in nature, such an extraction behavior of sol-gel methyl coating toward polar analytes is counterintuitive. Thus, sol-gel sorbents with short alkyl side chains have the potential to offer a polymer-free alternative to traditional sol-gel capillary microextraction (CME) media commonly prepared with the use of polymers in the sol solution. Elimination of polymers from the sol-gel coating solution is conducive to improving thermal stability and solvent tolerance of the created sol-gel extracting phase. This also makes the preparation of sol-gel coatings facile and cost-effective. Possessing excellent solvent stability, such sol-gel coatings offer the opportunity for effective on-line hyphenation of capillary microextraction with HPLC and other liquid-phase separation techniques that employ organo-aqueous mobile phases. PMID- 18420216 TI - Application of liquid-liquid-liquid microextraction and ion-pair liquid chromatography coupled with photodiode array detection for the determination of chlorophenols in water. AB - A method termed as liquid-liquid-liquid microextraction was utilized to extract chlorophenols from water. The extracted chlorophenols, present in anionic form, were then separated, identified, and quantitated by ion-pair high-performance liquid chromatography with photodiode array detection (HPLC/DAD). For trace chlorophenol determination using HPLC/DAD, the chlorophenolate anion provides a better ultraviolet spectrum for quantitative and qualitative analyses than does uncharged chlorophenol. This is due to the auxochromic effect of the phenolate anion. In the study, experimental conditions such as organic phase identity, acceptor phase volume, sample agitation, extraction time, acceptor phase NaOH concentration, donor phase HCl concentration, salt addition, and UV absorption wavelength were optimized. Relative standard deviations (RSD, 2.3-5.4%), coefficients of determination (r2 0.9994-0.9999), and detection limits (0.049 0.081 ng mL(-1)) of the proposed method were investigated under the selected conditions. The method was successfully applied to analyses of reservoir and tap water samples, and the relative recoveries of chlorophenols from the spiked reservoir and tap water samples were 94.1-100.4% and 87.8-101.2%, respectively. The proposed method is capable of identifying and quantitating each analyte to 0.5 ng mL(-1), confirming the HPLC/DAD technique to be quite robust for monitoring trace levels of chlorophenols in water samples. PMID- 18420217 TI - Stable aqueous film coating dispersion of zein. AB - The effects of plasticizers, pH, and electrolytes on film formation and physical stability of aqueous film coating dispersions (pseudolatexes) of zein were evaluated. The influence of plasticizer on film formation mechanism and minimum film-formation temperature (MFT) were monitored by means of hot stage microscopy (HSM). Furthermore, the effects of pH and electrolytes on the short-term physical stability of pseudolatexes were investigated by measuring relative absorbance, zeta potential, and particle size of the dispersions. With aqueous coating dispersions of zein, stages of film formation were identified. The dispersions plasticized with 20% (w/w) PEG 400 or glycerol formed mechanically strong and flexible films with the lowest glass transition temperature (T(g)). Physical stability of the aqueous zein dispersions was dependent on both pH and electrolyte content. At a pH ranging from 3 to 4, the aqueous dispersions of zein were stable for at least 2 months exhibiting the highest values for zeta potential, the smallest particle size, and a low volume of aggregates. The stable dispersion could be obtained containing a lower concentration of electrolytes (e.g., 10(-5) M). The physical stability of aqueous zein dispersions can be determined by the combined measurements of relative absorbance, zeta potential, and particle size. PMID- 18420218 TI - Diffusiophoresis and electrophoresis of a charged sphere perpendicular to two plane walls. AB - The problem of diffusiophoretic and electrophoretic motions of a dielectric spherical particle in an electrolyte solution situated at an arbitrary position between two infinite parallel plane walls is studied theoretically in the quasisteady limit of negligible Peclet and Reynolds numbers. The applied electrolyte concentration gradient or electric field is uniform and perpendicular to the plane walls. The electric double layer at the particle surface is assumed to be thin relative to the particle radius and to the particle-wall gap widths, but the polarization effect of the diffuse ions in the double layer is incorporated. To solve the conservative equations, the general solution is constructed from the fundamental solutions in both cylindrical and spherical coordinates. The boundary conditions are enforced first at the plane walls by the Hankel transforms and then on the particle surface by a collocation technique. Numerical results for the diffusiophoretic and electrophoretic velocities of the particle relative to those of a particle under identical conditions in an unbounded solution are presented for various cases. The collocation results agree well with the approximate analytical solutions obtained by using a method of reflections. The presence of the walls can reduce or enhance the particle velocity, depending on the properties of the particle-solution system and the relative particle-wall separation distances. The boundary effects on diffusiophoresis and electrophoresis of a particle normal to two plane walls are found to be quite significant and complicated, and generally stronger than those parallel to the confining walls. PMID- 18420219 TI - Adsorption of bulky molecules of nonylphenol ethoxylate on ordered mesoporous carbons. AB - Ordered mesoporous carbons (OMCs) with varying pore sizes were prepared using ordered mesoporous silica SBA-15 as hard templates. The OMCs possess abundant mesopores with narrow pore size distribution, on which the adsorption behavior of bulky molecules of nonylphenol ethoxylate (NPE) were investigated. The isotherms of NPE on OMCs can be fitted by Langmuir adsorption model, evidenced by the adsorption data. The surface area of the pores larger than 1.5 nm is a crucial factor to the adsorption capacity of NPE, whereas the most probable pore diameter of OMCs is crucial to the adsorption rate of NPE. The adsorption temperature has more significant effects on adsorption rate than the adsorption capacity. Theoretical studies show that the adsorption kinetics of NPE on OMCs can be depicted with the pseudo-second-order kinetic model. In addition, thermodynamic parameters of adsorption were evaluated based on the equilibrium constants related to the equilibrium of adsorption at different temperatures. PMID- 18420220 TI - Fabrication and characteristics of organic semiconductor nanoparticles of copper phthalocyanine oligomers. AB - Nanoparticles of copper phthalocyanine oligomers (O-CuPc) with peripheral carboxylic acid groups have successfully been prepared by a simple method of liquid phase direct precipitation in the presence of different surfactants. X-ray diffraction patterns, transmission electron microscopy, and UV-visible spectra are employed to characterize the novel organic nanoparticles. The sizes and size distribution of the resulting O-CuPc nanoparticles show a noticeable dependence on surfactants. Nonionic surfactant is helpful in forming uniform nanoparticles. Also we observe a remarkable nanosize effect of the O-CuPc particles. PMID- 18420221 TI - Preparation of hydrophilic/hydrophobic porous materials. AB - A novel porous material was designed and prepared in this work. A hydrophobic open-celled porous polystyrene (PS) was first synthesized via a concentrated emulsion polymerization of water in styrene. Subsequently the porous polystyrene was saturated with an aqueous solution of acrylamide (AM) and an initiator, which was subjected to another polymerization and the resulted polyacrylamide (PAM) penetrated in the cells and intercellular pores of the PS matrix. The PAM would change its volume according to the environmental humidity and thus adjusted the permeation of the material. The morphology, pore size distributions, water absorption, and vapor permeation of the materials were investigated. PMID- 18420222 TI - NO sensing in Pseudomonas aeruginosa: structure of the transcriptional regulator DNR. AB - All denitrifying bacteria can keep the steady-state concentrations of nitrite and nitric oxide (NO) below cytotoxic levels, controlling the expression of the denitrification gene clusters by redox signaling, mainly through transcriptional regulators belonging either to the DNR (dissimilative nitrate respiration regulator) or to the NnrR (nitrite and nitric oxide reductase regulator) subgroups of the FNR (fumarate and nitrate reductase regulatory protein)-CRP (cAMP receptor protein) superfamily. The NO dependence of the transcriptional activity of promoters regulated by these transcription factors has suggested that they may act as NO sensors in vivo. Despite great interest in the regulation of denitrification, which in Pseudomonas aeruginosa is strictly related to virulence, functional and structural characterization of these NO sensors is still lacking. Here we present the three-dimensional structure of the sensor domain of the DNR from P. aeruginosa at 2.1 A resolution. This is the first structure of a putative NO-sensing bacterial transcriptional regulator and reveals the presence of a large hydrophobic cavity that may be the cofactor binding site. Parallel spectroscopic evidence indicates that apo-DNR binds heme in vitro and that the heme-bound form reacts with carbon monoxide and NO, thus supporting the hypothesis that NO sensing involves gas binding to the ferrous heme. Preliminary experiments indicate that heterologous expression of the heme containing DNR yields a protein able to bind DNA in vitro. PMID- 18420223 TI - Hypoxia impairs embryo development and survival in black bream (Acanthopagrus butcheri). AB - Coastal environments are threatened by the increasing frequency, extent and severity of hypoxic events. Hypoxia affects vast areas around the world and often causes fish kills, reduced abundance, altered distribution, low benthic biomass and declines in fisheries. In Australia, many fisheries are based on sparid fishes and in the southern states black bream (Acanthopagrus butcheri) is important to both the recreational and commercial sectors. This species completes its entire life cycle in estuaries and annual recruitment is highly variable and very likely influenced by environmental conditions during the spawning season. In a laboratory-based experiment, fertilised black bream eggs (embryos) were exposed to five different levels of dissolved oxygen (DO). The DO levels were maintained in small test wells using nitrogen gas in a novel chamber design. Embryo development was assessed over a 2-day period and hatched larvae were observed until Day 2 post-hatch. Significant differences (p<0.05) were observed in embryonic development and survival as a function of DO level. In severely hypoxic conditions (30% saturation) survival to 1 day was reduced and no hatching occurred. In moderately hypoxic conditions (45-55%S), both precocious and delayed hatching was observed and hatch rates were reduced, whilst the number of hatched larvae with deformities increased, resulting in reduced larval lengths. No larvae survived to Day 2 post-hatch when held in hypoxic conditions (<55%S). This study demonstrates the detrimental effect that severe hypoxia can have on the early development of black bream which could result in reduced recruitment and lowered abundance. Other species that share similar early life histories may also be at risk. PMID- 18420224 TI - The use of selected genotoxicity assays in green-lipped mussels (Perna viridis): a validation study in Hong Kong coastal waters. AB - In Hong Kong, discharges of domestic and industrial wastewaters and disposal of contaminated muds into coastal waters have resulted in high levels of persistent organic contaminants in the water column, biota and bottom sediments. Although the acute toxicity of effluents has been tested by government regulators using mortality-based bioassays, the sublethal effects of many of the pollutants, including environmental carcinogens, have not been well studied. The objective of the present work was to examine the efficiency and efficacy of using: (1) DNA strand breaks (comet assay and alkaline agarose gel electrophoresis assay); (2) oxidative DNA damage (levels of 8-hydroxy-2-deoxyguanosine in cellular DNA) and (3) micronuclei (MN) formation as early warning toxicological indicators of potential environmental carcinogen exposure. Green-lipped mussels (Perna viridis) were transplanted to study sites and retrieved after 4, 8, 12, 16 and 30 days. The bioaccumulation of organic pollutants, including the polycyclic aromatic hydrocarbons (PAHs), organochlorine pesticides (OC pesticides) and polychlorinated biphenyls (PCBs) was observed, and compared with biomarker assays. Some significant dose- and time-dependent inductions of the genotoxic biomarkers, including 8-OHdG, DNA strand breaks and micronuclei were observed. In particular, several very strong (r>0.9) and significantly positive relationships were found at all sites during days 0-12. In addition, significant strong relationships between certain organic contaminants (carcinogenic PAHs and 4 common OC pesticides) and micronuclei were detected at the polluted sites, Tsim Sha Tsui and Sai Wan Ho. The results suggest that the use of micronuclei as a biomarker is potentially effective, especially at the more polluted sites. Nonetheless, further research is required to validate their effectiveness under varying field conditions, especially to elucidate responses under lower contaminant loads, interactions between the environmental and physiological factors affecting organisms in differing coastal environments, and changes in biotransformation and DNA repair enzymes in test organisms. PMID- 18420225 TI - Photocatalytic degradation of spill oils on TiO(2) nanotube thin films. AB - The nitrogen-doped TiO(2) nanotube (N-TNT) thin films were synthesized using ZnO nanorods as the template and doped with urea at 623K. Under ultraviolet (UV) and visible light irradiation, the efficiencies for photocatalytic degradation of methylene blue is as high as 30%. About 10% of toluene (representing aromatics in the spill oils) in sea water can be photocatalytically degraded under visible light radiation for 120 min. The aliphatic model compound (1-hexadecene) has, on the contrary, a less efficiency (8%) on the N-TNT photocatalyst. On the average, under visible light radiation, the effectnesses of the N-TNT for photocatalytic degradation of model compounds in the spill oils in sea water are 0.38 mg toluene/gN-TNTh and 0.25 mg 1-hexadecene/gN-TNTh. It is expected that spill oils in the harbors or seashores can be adsorbed and photocatalytically degraded on the N-TNT thin films that are coated onto levee at the sea water surface level. PMID- 18420226 TI - A nonlinear model with competition between prostate tumor cells and its application to intermittent androgen suppression therapy of prostate cancer. AB - The recurrence of a tumor is a crucial problem in hormonal therapy of prostate cancer. Recent studies suggest that intermittent androgen suppression administration may prolong or hopefully prevent the progression to the recurrence. It was shown that a simple mathematical model is useful to understand how and why intermittent administration can be effective and to seek a better medication scheme. In this paper, we propose a new model for the intermittent androgen suppression therapy. What is central in the new model is that the competitive effect between androgen-dependent and independent cancer cells is assumed to be essential for the decrease of androgen-independent cells under a normal androgen level. In the newly proposed model, the separatrix in the phase space for a normal androgen level plays an important role. There is crucial difference between the previous model and the new one in the phase diagram of success and failure of the permanent tumor control by intermittent androgen suppression administration. That means that the suggestions from the models for clinical problems can be different. We also consider the combined model of the previous and newly proposed models, which can smoothly bridge two models. PMID- 18420227 TI - [The concept of an epileptogenic network in human partial epilepsies]. AB - An anatomical and functional model of drug-resistant partial seizures is presented and discussed based on research conducted by our team over the last decade. This research is based on the study of intracerebral stereoelectroencephalography (SEEG) recordings in an attempt to identify the neural networks involved in generating paroxystic activities so as to understand their dynamics in space and time, and to propose targeted therapies that could "control" these networks. Today, the classical notion of epileptic focus should be replaced by a more complex model that takes into account the potential interactions within the neuronal networks involved in seizures. During partial epileptic seizures, the cerebral structures involved are the seat of characteristic oscillations that may be synchronized or, on the contrary, that can desynchronize in a transitory manner. These epileptic rhythms disturb the physiological rhythms that underlie the cognitive and emotional processes, which can thus be altered in partial epilepsy, even if located far from the original discharge site. We suggest that seizures originate in a group of structures that are highly epileptogenic (epileptogenic zone network, [EZN]) whose activity is synchronized before the appearance of fast oscillations that are transitorily desynchronized. Later, other cortical and subcortical structures are the seat of slower, synchronized rhythmic modifications (propagation network, [PN]). The emergence of clinical signs in the seizure depend on these phenomena, which in some cases can mimic a normal cognitive process or, on the contrary, lead to a deep rupture in normal cerebral functioning. PMID- 18420228 TI - [Does epilepsy surgery really lower mortality?]. AB - Patients with epilepsy suffer from a two to three fold increased death rate as compared to age and sex matched control population. This increased risk culminate to five fold in patients with drug resistant partial epilepsy eligible for epilepsy surgery, with the majority of deaths classified as sudden unexpected death in epilepsy (SUDEP). The pathophysiology of SUDEP remains uncertain, but all witnessed cases occurred during or immediately after a seizure. Several studies have evaluated the impact of epilepsy surgery on the risk of seizure related death and SUDEP. Four series have concentrated on operated patients, and have compared the death rates in those seizure free and non seizure free post operatively. Three of these studies reported a significantly lower risk of SUDEP in patients cured by surgery as compared to those still seizing. Four other series have compared the mortality in surgically versus medically treated patients with refractory partial epilepsy. Three of these studies failed to show any significant difference in death or SUDEP rates between operated and and non operated patients. All the above series suffer various types of methological limitations, hampering any definite conclusion regarding the impact of epilepsy surgery on mortality. The launching of novel and large multicentric studies, which address the pitfalls of prior series, should allow to provide conclusive results within the next three years. PMID- 18420229 TI - [Epileptiform activities generated in vitro by human temporal lobe tissue]. AB - Drug-resistant partial epilepsies, including temporal lobe epilepsies with hippocampal sclerosis and cortical dysplasias, offer the opportunity to study human epileptic activity in vitro since the preferred therapy often consists of the surgical removal of the epileptogenic zone. Slices of this tissue retain functional neuronal networks and may generate epileptic activity. The properties of cells in this tissue do not seem to be significantly changed, but excitatory synaptic characteristics are enhanced and GABAergic inhibition is preserved. Typically, epileptic activity is not generated spontaneously by the neocortex, whether dysplastic or not, but can be induced by convulsants. The initiation of ictal discharges in neocortex depends on both GABAergic signaling and increased extracellular potassium. In contrast, a spontaneous interictal-like activity is generated by tissues from patients with temporal lobe epilepsies associated with hippocampal sclerosis. This activity is initiated not in the hippocampus but in the subiculum, an output region that projects to the entorhinal cortex. Interictal events seem to be triggered by GABAergic cells, which paradoxically excite approximately 20% of subicular pyramidal cells, while simultaneously inhibiting the majority. Interictal discharges are therefore sustained by both GABAergic and glutamatergic signaling. The atypical depolarizing effects of GABA depend on a pathological elevation in the basal levels of chloride in some subicular cells, similar to those of developmentally immature cells. This defect is caused by the perturbation of the expression of the cotransporters regulating the intracellular chloride concentration, the importer NKCC1, and the extruder KCC2. Blockade of excessive NKCC1 by the diuretic bumetanide restores intracellular chloride and thus hyperpolarizing GABAergic actions, suppressing interictal activity. PMID- 18420230 TI - [Predictors of verbal memory decline following temporal lobe surgery]. AB - BACKGROUND AND PURPOSE: Verbal memory decline can occur after temporal lobe surgery, especially when the left dominant hemisphere is involved. This potential functional risk must be evaluated before surgery. Among all factors that have been identified by several studies, the side of surgery (left dominant) and high baseline memory performance have been found to be predictive of verbal memory decline. Other factors such as etiology, sex, age at surgery, age at seizure onset, and duration may influence memory decline, but the results are not clear. Our purpose was to identify, in our population of patients and among all risk factors, those that may be predictive of verbal memory decline. METHODS: Logistic regression was used to examine the effect of each factor on the postoperative verbal memory index (WMS-R) in 101 patients who underwent a right (n=49) or left (n=52) anterior temporal lobe resection. RESULTS: In the group as a whole, 22 % of the patients demonstrated verbal memory decline of more than one standard deviation. The verbal memory decline was significantly related to surgery on the left side and a high level of verbal memory performance. These factors were significant predictors of decline. The other factors (etiology, sex, age at surgery, age at seizure onset, and duration) were not found to be predictive of this decline. CONCLUSIONS: Our analysis demonstrates that the patients who are most at risk of undergoing verbal memory deterioration are those who undergo left sided temporal resection and have good memory scores preoperatively. The contradictions found in the literature about the other factors could be explained by the diversity of the tests and criteria used to assess memory decline. PMID- 18420231 TI - [Corticectomy: technical considerations]. AB - The surgical treatment of epilepsy requires careful preparation and presents a certain number of technical specificities. The neurosurgeon must master not only the technical aspects but also the therapeutic and functional trade-off in order to modulate the procedure according to morphological and electrophysiological intraoperative data. A large number of technical variants have been developed to correspond to epileptological or functional anatomical considerations. Until this point, the choice of a particular technique does not seem to have a significant impact on the therapeutic effectiveness of surgery, and differences in results can be related to the presurgical evaluation and surgical indications. On the other hand, technical development promises to play an important role in limiting the long-term neurocognitive consequences of surgery. PMID- 18420232 TI - Interhemispheric interaction during the menstrual cycle. AB - Fluctuating levels of sex hormones and high levels of progesterone (P), in particular, have been suggested to reduce interhemispheric inhibition. The present study focuses on hormone-dependent modulation of interhemispheric integration. In two versions of the Banich-Belger task, participants were asked to match letters according to their physical (e.g., A vs. A) and semantic identity (e.g., A vs. a). Matches were presented either within or across visual half-fields. Moreover, a simple reaction-time task (Poffenberger task) that is assumed to estimate interhemispheric transfer time (IHTT) was used. Seventeen normally cycling women were tested during low P menses and high P midluteal phase. Saliva levels of P were analysed using chemiluminescence assays. Fifteen postmenopausal women performed the same tasks in corresponding time intervals. Additionally, 28 younger male controls were tested once. In agreement with previous results, the more demanding (semantic) interhemispheric-integration task revealed a typical across-field advantage (AFA) for all three groups. However, in normally cycling women, the AFA was significantly reduced during menses. IHTT did not change across the cycle phases. The results indicate that interhemispheric integration fluctuates across the menstrual cycle and is reduced during menses. During the luteal phase, however, the AFA is increased, suggesting that accompanying hormonal conditions favour an efficient interhemispheric integration. We conclude that transcallosal mechanisms involved in interhemispheric integration are profoundly altered when sex hormones are permanently reduced as in men and postmenopausal women. This difference enables an efficient interhemispheric integration without modulatory effects of P. PMID- 18420233 TI - Letter position coding in attentional dyslexia. AB - We report data from experiments on orthographic and phonological coding in two patients with attentional dyslexia following bilateral parietal damage. Two experiments required the patients to carry out lexical decisions and we varied whether the nonwords were orthographically or phonologically similar to real words. Experiment 1 showed that the patients were sensitive to the orthographic relations between nonwords and words, as they tended to accept as words nonwords whose letters could migrate within the string to form a word. There were no effects of phonological similarity between the nonwords and the words. Experiment 2 demonstrated that the patients were less likely to accept nonwords as words if the word had to be formed by transposing the first two letters in the string. The data suggest that attentional dyslexics are primarily sensitive to orthographic similarity between words and nonwords, and also that the first letters have privileged coding of their locations, despite the patients being poor at coding letter positions. The implications for theories of visual word recognition are discussed. PMID- 18420234 TI - Hippocampal activation during episodic and semantic memory retrieval: comparing category production and category cued recall. AB - Whether or not the hippocampus participates in semantic memory retrieval has been the focus of much debate in the literature. However, few neuroimaging studies have directly compared hippocampal activation during semantic and episodic retrieval tasks that are well matched in all respects other than the source of the retrieved information. In Experiment 1, we compared hippocampal fMRI activation during a classic semantic memory task, category production, and an episodic version of the same task, category cued recall. Left hippocampal activation was observed in both episodic and semantic conditions, although other regions of the brain clearly distinguished the two tasks. Interestingly, participants reported using retrieval strategies during the semantic retrieval task that relied on autobiographical and spatial information; for example, visualizing themselves in their kitchen while producing items for the category kitchen utensils. In Experiment 2, we considered whether the use of these spatial and autobiographical retrieval strategies could have accounted for the hippocampal activation observed in Experiment 1. Categories were presented that elicited one of three retrieval strategy types, autobiographical and spatial, autobiographical and nonspatial, and neither autobiographical nor spatial. Once again, similar hippocampal activation was observed for all three category types, regardless of the inclusion of spatial or autobiographical content. We conclude that the distinction between semantic and episodic memory is more complex than classic memory models suggest. PMID- 18420235 TI - The effects of prolonged caloric restriction leading to weight-loss on food hedonics and reinforcement. AB - Evidence suggests that when animals are chronically deprived of calories, the reward of a food stimulus becomes more salient. Recently leptin has been implicated in food reward. The objective of this study was to examine the effect of weight-loss on food hedonics and on the reinforcing value of palatable snack food, and to determine whether plasma leptin concentrations were related to these variables. Fifteen apparently healthy obese adults (n=9 women and 6 men; age=33.5+/-7.8; BMI=35.7+/-1.1 kg/m(2)) were subjected to 8 weeks of caloric deprivation (-700 kcal/day). Plasma leptin (ELISA), body weight and composition (Dual X-ray Absorptiometry), food reinforcement (concurrent schedules task) and food hedonics (visual analogue scale) were measured pre- and post-intervention. Body weight decreased by 5.2+/-2.7% (p=0.001) while leptin decreased by 30.1+/ 32.6% (p=0.001). Relative to baseline, after weight-loss food hedonics or "liking" was rated significantly higher for the food reinforcers (p=0.001) offered at sessions end, and this change was not significantly correlated with changes in plasma leptin. No significant effect of weight-loss was noted for the reinforcing value of palatable snack food. In conclusion, after 8 weeks of caloric deprivation there was no change in the reinforcing value of palatable snack foods, but the rating of food "liking" increased ~10%, and this increase was independent of weight-loss. PMID- 18420236 TI - Recall of recent lunch and its effect on subsequent snack intake. AB - Recall of food eaten at lunch decreases afternoon snack intake, suggesting that awareness of a recent eating episode may be an important factor influencing appetite. The aim of the present studies was to investigate whether the inhibitory effect of remembering a recent lunch meal on subsequent snack intake is dependent upon 1) the type and palatability of the snack offered; 2) participants' level of dietary restraint and tendency towards disinhibition; and 3) the delay between lunch and recall. Popcorn snacks differing in amount of added salt and rated palatability were offered to male participants in Experiment 1. Participants who recalled the lunch they had eaten that day consumed less of all types of popcorn than participants who recalled lunch eaten the previous day, suggesting that the effect of recent meal recall is not dependent upon the palatability of the snack food. In Experiment 2, a similar pattern of results was observed but only for women who scored low, and not high, on a measure of tendency toward dietary disinhibition, possibly because a tendency toward disinhibition is associated with impaired memory for the lunch. In Experiment 3, decreased cookie intake by women was observed after remembering today's lunch relative to a neutral control condition, but this effect was similarly only observed for participants scoring low in tendency toward disinhibition. In addition, the effect was dependent on the time elapsed between the lunch and recall, since intake was only reduced at a snack tasting session 3-hours post lunch (when some forgetting of the meal occurred) and not 1-hour post-lunch. It is concluded that the inhibitory effect of recalling foods eaten at lunch on subsequent snack intake is a robust phenomenon that is related to memory of that lunch and is moderated by tendency toward dietary disinhibition. PMID- 18420237 TI - Eudesmanes from Pluchea sagittalis. Their antifeedant activity on Spodoptera frugiperda. AB - Eudesmane-type sesquiterpenoids 3alpha-(2,3-epoxy-2-methylbutyryloxy)-4alpha formoxy-11-hydroxy-6,7-dehydroeudesman-8-one (1) and 3alpha-(2,3-epoxy-2 methylbutyryloxy)-4alpha,7alpha,11-trihydroxyeudesman-8-one (2), together with 10 known structurally related eudesmanes were isolated from the CHCl3 extract of aerial parts of Pluchea sagittalis (Lamarck) Cabrera. Their structures were deduced by extensive application of 1 and 2D NMR spectroscopic techniques and high and low resolution CIMS. X-ray crystallographic analysis of the known compound 3alpha-(2,3-epoxy-2-methylbutyryloxy)-4alpha-formoxycuauthemone (9) is reported here for the first time, and confirms the structural features for the series of the reported eudesmanes. All eudesmanes were tested for their antifeedant activity by incorporating them to an artificial diet of larvae of the polyphagous insect Spodoptera frugiperda at a concentration of 100ppm. Our results, from feeding choice tests, indicated that most of the compounds deter larval feeding at the cited concentration. PMID- 18420238 TI - Differences in the influence of tobacco smoking on lung cancer between Japan and the USA: possible explanations for the 'smoking paradox' in Japan. AB - OBJECTIVES: The prevalence of cigarette smoking among Japanese men has been consistently high compared with males in Western countries over the past 30 years. However, during the same period, the incidence and mortality rates for lung cancer have been consistently lower in Japan than in Western countries, which has been termed the 'Japanese smoking paradox'. The odds ratio/relative risk of cigarette smoking for lung cancer mortality and incidence for the same number of cigarettes smoked per capita in Japan have been lower than those in Western countries. This difference in the odds ratio/relative risk is likely to be the main reason for the Japanese smoking paradox. The aim of this study was to clarify the reason for the difference in the odds ratio/relative risk between Japan and the USA. STUDY DESIGN: Literature review to compare environmental, hereditary and other factors that may be related to lung cancer in Japan and the USA. RESULTS AND CONCLUSIONS: The main factors likely to have brought about the difference in the odds ratio/relative risk between Japan and the USA (and perhaps other Western countries as well) are: lower alcohol consumption by Japanese males; lower fat intake by Japanese males; higher efficiency of filters on Japanese cigarettes; lower levels of carcinogenic ingredients in Japanese cigarettes; and lung-cancer-resistant hereditary factors among Japanese males. PMID- 18420239 TI - Dengue haemorrhagic fever integral hypothesis: confirming observations, 1987 2007. AB - In 1987, Kouri et al. published in Transactions their integral hypothesis to explain the development of dengue haemorrhagic fever (DHF) epidemics (Kouri, G.P., Guzman, M.G., Bravo, J.R., 1987. Why dengue haemorrhagic fever in Cuba? 2. An integral analysis. Trans. R. Soc. Trop. Med. Hyg. 81, 821-823). Based on observations carried out during the 1981 Cuban DHF epidemic, the authors integrated in one model the most advanced knowledge at that time. Observations in the last 20 years confirm the importance of this multifactorial and unifying view of the problem, where the interaction between the host, the virus and the vector in an epidemiological and ecosystem setting might determine DHF as a final outcome. Investigations on the interaction among host, virus and mosquito with an ecosystemic view are needed. PMID- 18420240 TI - Mechanistic aspects of rearrangement of 16alpha-hydroxy-17-keto steroids to the 17beta-hydroxy-16-keto isomers. AB - The mechanistic aspects of the alkali-catalyzed rearrangement of 16alpha-hydroxy 17-keto steroid 1 to 17beta-hydroxy-16-keto steroid 2 are elucidated by use of (18)O- and deuterium-labeling experiments. The (18)O-labeling experiments refute the gem-hydration-quasi-diaxial dehydration mechanism for the rearrangement previously proposed and support the conventional enolization mechanism. Moreover, equilibrium by gem-hydration-dehydration occurs at the C-17 carbonyl more efficiently than at the C-16 carbonyl. Enolization rate of a carbonyl group at C 16 of 17beta-ketol 2 toward the C-17 position (k(16,17)) was about 8-10 times higher than those of 16alpha-ketol 1 toward the C-16 position (k(17,16)) and ketol 2 toward the C-15 position (k(16,15)). The marked deuterium-isotope effect on each enolization was observed with k(H)/k(D) ranging between 5.4 and 8.8. The present findings reveal that the initial hydration-dehydration equilibration at the C-17 carbonyl of ketol 1 followed by enolization of the carbonyl gives the ene-diol intermediate that isomerizes quantitatively to the 16-keto isomer of which the 16-carbonyl moiety enolizes preferentially toward the C-17 position rather than the C-15 position, yielding the ene-diol. Computational calculations of ground state energies of ketols 1-M and 2-M, trans-cyclohexane/cyclopentane structures, and their activation energies in the rearrangement support the dynamic aspects of the rearrangement as well as the kinetics data of the enolization. PMID- 18420241 TI - The co-evolution of culturally inherited altruistic helping and cultural transmission under random group formation. AB - Limited migration results in kin selective pressure on helping behaviors under a wide range of ecological, demographic and life-history situations. However, such genetically determined altruistic helping can evolve only when migration is not too strong and group size is not too large. Cultural inheritance of helping behaviors may allow altruistic helping to evolve in groups of larger size because cultural transmission has the potential to markedly decrease the variance within groups and augment the variance between groups. Here, we study the co-evolution of culturally inherited altruistic helping behaviors and two alternative cultural transmission rules for such behaviors. We find that conformist transmission, where individuals within groups tend to copy prevalent cultural variants (e.g., beliefs or values), has a strong adverse effect on the evolution of culturally inherited helping traits. This finding is at variance with the commonly held view that conformist transmission is a crucial factor favoring the evolution of altruistic helping in humans. By contrast, we find that under one-to-many transmission, where individuals within groups tend to copy a "leader" (or teacher), altruistic helping can evolve in groups of any size, although the cultural transmission rule itself hitchhikes rather weakly with a selected helping trait. Our results suggest that culturally determined helping behaviors are more likely to be driven by "leaders" than by popularity, but the emergence and stability of the cultural transmission rules themselves should be driven by some extrinsic factors. PMID- 18420242 TI - Activation of oxidative stress-responsive signaling pathways in early splenotoxic response of aniline. AB - Aniline exposure causes toxicity to the spleen, which leads to a variety of sarcomas, and fibrosis appears to be an important preneoplastic lesion. However, early molecular mechanisms in aniline-induced toxicity to the spleen are not known. Previously, we have shown that aniline exposure results in iron overload and induction of oxidative stress in the spleen, which can cause transcriptional upregulation of fibrogenic/inflammatory cytokines via activation of oxidative stress (OS)-responsive signaling pathways. To test this mechanism, male SD rats were treated with aniline (1mmol/kg/day via gavage) for 7 days, an experimental condition that precedes the appearance of fibrosis. Significant increases in both NF-kappaB and AP-1 binding activity was observed in the nuclear extracts of splenocytes from aniline-treated rats as determined by ELISAs, and supported by Western blot data showing increases in p-IkappaBalpha, p-p65 and p-c-Jun. To understand the upstream signaling events which could account for the activation of NF-kappaB and AP-1, phosphorylation patterns of IkappaB kinases (IKKalpha and IKKbeta) and mitogen-activated protein kinases (MAPKs) were pursued. Our data showed remarkable increases in both p-IKKalpha and p-IKKbeta in the splenocytes from aniline-treated rats, suggesting their role in the phosphorylation of both IkappaBalpha and p65 subunits. Furthermore, aniline exposure led to activation of all three classes of MAPKs, as evident from increased phosphorylation of extracellular-signal-regulated kinase (ERK1/2), c-Jun N-terminal kinase (JNK1/2) and p38 MAPKs, which could potentially contribute to the observed activation of both AP-1 and NF-kappaB. Activation of upstream signaling molecules was also associated with simultaneous increases in gene transcription of cytokines IL-1, IL-6 and TNF-alpha. The observed sequence of events following aniline exposure could initiate a fibrogenic and/or tumorigenic response in the spleen. PMID- 18420243 TI - Intracellular localization and subsequent redistribution of metal transporters in a rat choroid plexus model following exposure to manganese or iron. AB - Confocal microscopy was used to investigate the effects of manganese (Mn) and iron (Fe) exposure on the subcellular distribution of metal transporting proteins, i.e., divalent metal transporter 1 (DMT1), metal transporter protein 1 (MTP1), and transferrin receptor (TfR), in the rat intact choroid plexus which comprises the blood-cerebrospinal fluid barrier. In control tissue, DMT1 was concentrated below the apical epithelial membrane, MTP1 was diffuse within the cytosol, and TfR was distributed in vesicles around nuclei. Following Mn or Fe treatment (1 and 10 microM), the distribution of DMT1 was not affected. However, MTP1 and TfR moved markedly toward the apical pole of the cells. These shifts were abolished when microtubules were disrupted. Quantitative RT-PCR and Western blot analyses revealed a significant increase in mRNA and protein levels of TfR but not DMT1 and MTP1 after Mn exposure. These results suggest that early events in the tissue response to Mn or Fe exposure involve microtubule-dependent, intracellular trafficking of MTP1 and TfR. The intracellular trafficking of metal transporters in the choroid plexus following Mn exposure may partially contribute to Mn-induced disruption in Fe homeostasis in the cerebrospinal fluid (CSF) following Mn exposure. PMID- 18420244 TI - MALDI-TOF-MS detection of the low molecular weight neurotoxins anatoxin-a and homoanatoxin-a on lyophilized and fresh filaments of axenic Oscillatoria strains. AB - Anatoxin-a (ANTX) and homoanatoxin-a (HANTX) are low molecular weight neurotoxic secondary amines of 165 and 179 Da, respectively. We applied matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS) for the detection of ANTX and HANTX directly on lyophilized and fresh filaments of axenic strains of the genus Oscillatoria, using 2,5-dihydroxybenzoic acid as matrix and purified ANTX and HANTX as references. To counteract the span of low molecular mass ions (< m/z 1000) generated by the matrix, we induced the matrix suppression effect to obtain high quality ANTX/HANTX MALDI signals. MALDI desorption/ionization of the matrix-ANTX and the matrix-HANTX generated protonated molecules [M+H](+) at m/z 166.12322 and 180.1372, respectively. The masses obtained from the analysis of lyophilized filaments of the ANTX-producer Oscillatoria sp. strain PCC 9240 (m/z 166.15) and of fresh filaments of the HANTX producers Oscillatoria sp. strains PCC 6506 (m/z 180.1375), PCC 9029 (m/z 180.1334) and PCC 10111 (m/z 180.13996) corresponded to the protonated molecular ions of ANTX and HANTX, respectively. Therefore, the application of MALDI-TOF-MS for the detection of cyanobacterial anatoxins in clonal and axenic strains of the cyanobacterial culture collections worldwide may help to assess ANTX/HANTX incidence among cyanobacteria. PMID- 18420245 TI - Accumulation of tetrodotoxin and 4,9-anhydrotetrodotoxin in cultured juvenile kusafugu Fugu niphobles by dietary administration of natural toxic komonfugu Fugu poecilonotus liver. AB - Non-toxic cultured juvenile kusafugu Fugu niphobles were fed with a diet containing highly toxic natural komonfugu Fugu poecilonotus liver until the 30th day (8.0 microg of TTX and 3.7 microg of 4,9-anhydroTTX/fish/day), and then fed with a non-toxic diet until the 240th day. During the 30-240th day, five or six fish were periodically sampled six times, and the contents of TTX and 4,9 anhydroTTX in each tissue were determined. The total TTX and 4,9-anhydroTTX accumulated in all tissues tested was not significantly changed during the experimental period, both being kept at 70% of administrated doses. However, in the liver, the TTX content accounted to be 120 microg (50% of administrated) on the 30th day, and then it gradually decreased to 50 microg until the 240th day, while 4,9-anhydroTTX content was kept at approximately 40 microg (40% of administrated) during all the experimental periods. In contrast to the liver, in the skin, TTX and 4,9-anhydroTTX were 40 and 5 microg, respectively, on the 30th day, and then gradually increased to 80 and 24 microg, respectively, until the 240th day. In the intestine, TTX and 4,9-anhydroTTX contents were kept at 25 and 12 microg, respectively, during all the experimental periods. According to these results, we assumed that a part of TTX accumulated in the liver was slowly transferred to the skin. PMID- 18420246 TI - Ochratoxin A in the morning and afternoon portions of urine from Coimbra and Valencian populations. AB - The widespread contamination of foodstuffs and beverages by mycotoxins, such as ochratoxin A (OTA), has made the monitoring of human contamination levels essential. By using a sensitive, accurate and speedy method that combines extraction with 5% NaHCO(3), immunoaffinity column clean-up and HPLC with fluorescence detection, the human exposure to OTA through urine analysis can be monitored. This method is less invasive than blood monitoring and has the potential to be a good marker of human exposure. The limit of quantification of the method was 0.007 ng/mL of urine, with recoveries of OTA, from urine samples spiked at levels between 0.02 and 0.1 ng/mL, higher than 91% with RSD lower than 15.5%. This study evaluated OTA contamination levels in human urine sample fractions, collected in the morning and afternoon, in two populations, one from Coimbra city, in Portugal, and another from the Valencian community, in Spain. In the Coimbra population, 60 samples from 30 healthy individuals were analyzed, levels of OTA in 13 morning samples and 14 afternoon samples having been detected, with concentrations ranging from 0.011 to 0.208 and 0.008 to 0.11 ng/mL respectively. In the Valencia population, 62 samples from 31 healthy individuals were analyzed, with OTA being detected in 25 morning samples and 26 afternoon samples. The concentrations varied between 0.007 and 0.124 ng/mL in the morning samples, and 0.008 and 0.089 ng/mL in the afternoon samples. Significant differences were found between the morning levels of OTA from both populations (P=0.033). For afternoon samples, significant differences were not found, P value=0.163. PMID- 18420247 TI - Characterization of variable genomic regions of Indian white spot syndrome virus. AB - A recent study examining genetic variations among the 3 completely sequenced white spot syndrome virus (WSSV) genomes isolated from China, Thailand and Taiwan revealed five major differences among them. Of these differences, a deletion region between ORF 23/24 and a variable region of ORF14/15 prone to recombination were of particular evolutionary significance. Focusing on these regions, 81 WSSV isolates from India were characterized by sequencing polymerase chain reaction (PCR) amplicons. The Indian strains carried a 10,970 bp deletion in the ORF 23/24 region relative to WSSV-TW and WSSV-TH-96-II. Analysis of the ORF 14/15 regions revealed two novel strains of WSSV with unique sequences which could have evolved by recombination. None of the WSSV isolates had a transposase sequence or VP35 gene as reported for Taiwan isolates. The Indian strains were closely related to Thailand strains suggesting movement of a putative ancestor from Thailand to other parts of the world including India. PMID- 18420248 TI - Severe seasonal influenza in ferrets correlates with reduced interferon and increased IL-6 induction. AB - Even though ferrets are one of the principal animal models for influenza pathogenesis, the lack of suitable immunological reagents has so far limited their use in host response studies. Using recently established real-time PCR assays for a panel of ferret cytokines, we analyzed the local ferret immune response to human influenza isolates of the H1N1 and H3N2 subtypes that varied in their virulence. We observed that the severity of clinical signs correlated with gross- and histopathological changes in the lungs and was subtype-independent. Strains causing a mild disease were associated with a strong and rapid innate response and upregulation of IL-8, while severe infections were characterized by a lesser induction of type I and II interferons and strong IL-6 upregulation. These findings suggest that more virulent strains may interfere more efficiently with the host response at early disease stages. PMID- 18420249 TI - Open reading frame 193R of Chilo iridescent virus encodes a functional inhibitor of apoptosis (IAP). AB - Programmed cell death or apoptosis is a major defense mechanism in insects in response to viral infections. The genome of Chilo iridescent virus (CIV) has three ORFs with homology to baculovirus inhibitor of apoptosis (iap) genes. The proteins encoded by the 157L, 193R, and 332L ORFs contain 152, 208 and 234 amino acids, respectively. While all three proteins contain C-terminal RING domains, only the protein encoded by ORF 193R contains a baculoviral iap repeat (BIR) domain, indicative of a putative IAP protein. The 193R protein has 28 and 27% similarity in amino acid sequence to the Orgyia pseudotsugata MNPV and Cydia pomonella granulovirus IAP-3 proteins, respectively. ORF 193R from CIV is the only gene known to exist among the Iridoviridae that encodes a BIR domain. 193R is transcribed early during CIV infection, and its transcription is not dependent on the synthesis of early viral proteins. When this putative CIV IAP was transiently expressed in SPC-BM-36 and Sf21 cells under the control of an immediate early baculovirus promoter it significantly reduced apoptosis induced by actinomycin-D. Silencing of the CIV iap gene (193R) in CIV infected SPC-BM-36 cells with 193R-specific dsRNA resulted in apoptosis. Thus, CIV ORF 193R is the first iap gene identified in an iridovirus, which encodes a functional IAP protein. PMID- 18420250 TI - Sorption kinetics of 2,4-D and carbaryl in selected agricultural soils of northern Iraq: application of a dual-rate model. AB - Agriculture in northern Iraq (Kurdistan) relies on the widespread use of pesticides to promote crop performance. Over-application of many pesticides is commonplace, however, and may compromise soil and water quality, and ultimately human health, within the region. The aim of this study was to investigate the sorption-desorption kinetics and equilibrium partitioning of two selected pesticides in agricultural soils from northern Iraq. This was achieved by fitting a dual-rate sorption-desorption model to time-dependent data obtained from batch experiments. 2,4-D and carbaryl were selected for scrutiny since both are in common use in the region. Six agricultural soils, sampled around the city of Erbil, were investigated. These were low in organic carbon (OC) compared with many agricultural soils from more temperate regions. However, there was still a clear trend of increasing sorption of both 2,4-D and carbaryl with increasing % OC. In the case of both compounds, fast and slow adsorption rate coefficients and 48 h experimental K(d) values were positively correlated with % OC. It was assumed that K(OC) would provide a simple and reliable predictor of K(d). However, while this assumption holds true for short-term (48 h) experimental data, longer-term sorption in some soils (as indicated by theoretical K(d) values estimated from kinetic parameters in our study) appears to be under-predicted by K(OC) alone. The data presented here provide a useful starting point for further site-specific investigations of pesticide impacts in the Kurdistan region of Iraq. PMID- 18420251 TI - Carbonyls and non-methane hydrocarbons at a rural mountain site in northeastern United States. AB - Measurements of carbonyls and C(2)-C(6) non-methane hydrocarbons (NMHCs) were made in ambient air at a rural site at the summit of Whiteface Mountain (WFM) in New York State. Alkanes dominated in the samples, with ethane and propane making up about 55% of the total on a carbon-atom basis. Ethane, the longest-lived of the NMHCs, showed a mixing ratio in the range of 0.86-2.1 ppbv. Photochemical ageing analysis indicated an anthropogenic influence on the NMHC levels. The photochemical reactivity of the hydrocarbons, calculated in terms of propylene equivalent concentration, was dominated by alkenes (propene and ethene), which accounted for 74% of the total NMHC sum. Air mass back-trajectories have been used to investigate the origin of the observed NMHCs and carbonyls. Higher concentrations were found when air masses arrived from the midwestern US corridor. Acetone was the most abundant species, comprising from 31% to 53% of the total detected carbonyls, followed by MEK (15-53%), HCHO (7-39%), and CH(3)CHO (7-19%). Average concentrations were determined to be 1.61 ppbv for CH(3)C(O)CH(3), 1.40 ppbv for MEK, 1.16 ppbv for HCHO, and 0.49 ppbv for CH(3)CHO. The variations in carbonyl concentrations were observed to follow patterns similar to variations in O(3) concentrations, typical of secondary products. Correlations and statistical analysis of the carbonyls and NMHCs were performed, and showed that most of the compounds derived from mixing and photochemical transformation of long-range transported pollutants from the major source areas. Ranking of the carbonyls with respect to removal of the OH radical showed HCHO to be the most important species, followed by CH(3)CHO, MEK, and CH(3)C(O)CH(3). PMID- 18420252 TI - Sublethal responses in caged organisms exposed to sediments affected by oil spills. AB - This study was performed to determine sublethal responses of two invertebrate species by using field deployments in areas affected by oil spills, which are acute in the Galician Coast (NNW, Spain) and chronic in the Bay of Algeciras (SSW, Spain). The organisms employed were the crab Carcinus maenas and the clam Ruditapes philippinarum, and during 28 days the animals were exposed to contaminated sediments in cages under field conditions. Different biomarkers of exposure were determined after a 28-day period exposure: ethoxyresorufin O deethylase (EROD), phase I detoxification enzyme, glutathione-S-transferase (GST) phase II detoxification enzyme but also implicated in oxidative stress events, glutathione peroxidase (GPX) and glutathione reductase (GR), both antioxidant enzymes. In addition, histopathological effects in target tissues of the deployed organisms were evaluated. Biomarker measurements were linked with the concentration of chemicals in the sediments in order to elucidate the type, source and bioavailability of contaminants that produce adverse effects in the bioindicator species. Results obtained in this study have shown how the application of the selected battery of biomarkers under field bioassays allows for the identification of alternative sources of stress that are not observable in laboratory experiments. PMID- 18420253 TI - DNA damage, DNA repair, ageing and age-related disease. PMID- 18420254 TI - Sublethal zinc exposure has a detrimental effect on reproductive performance but not on the cyst hatching success of Artemia parthenogenetica. AB - The sublethal zinc toxicity to Artemia parthenogenetica as regards the possibility of colonization of zinc polluted salterns by means of cysts has been assessed by a cyst hatching assay and a life table approach. Emergence and hatching at different times as well as the whole hatching profile were taken as end-points for evaluating success of development. Demographic and reproductive parameters calculated according to the Lotka equation were used as an indicator of the chronic toxicity of the population. No adverse effects of waterborne zinc were found on hatching and emergence of cysts of A. parthenogenetica at any of the concentrations tested (0.01 mg/l, 0.1 mg/l, 0.5 mg/l, 1 mg/l, 5 mg/l). Chronic zinc exposure at 0.08 mg/l had detrimental effects on A. parthenogenetica fecundity, as detected by a decrease in the percentage of fertile females, which in turn produces a decrease in r. Chronic toxicity of zinc may be a limiting step for A. parthenogenetica colonization and the establishing of permanent populations in zinc-polluted brine ponds. PMID- 18420255 TI - Occurrence and fate of emerging wastewater contaminants in Western Balkan Region. AB - This paper reports on a comprehensive reconnaissance of over seventy individual wastewater contaminants in the region of Western Balkan (WB; Bosnia and Herzegovina, Croatia and Serbia), including some prominent classes of emerging contaminants such as pharmaceuticals and personal care products, surfactants and their degradation products, plasticizers, pesticides, insect repellents, and flame retardants. All determinations were carried out using a multiresidue analytical approach, based on the application of gas chromatographic and liquid chromatographic techniques coupled to mass spectrometric detection. The results confirmed a widespread occurrence of the emerging contaminants in municipal wastewaters of the region. The most prominent contaminant classes, determined in municipal wastewaters, were those derived from aromatic surfactants, including linear alkylbenzene sulphonates (LAS) and alkylphenol polyethoxylates (APEO), with the concentrations in raw wastewater reaching into the mg/l range. All other contaminants were present in much lower concentrations, rarely exceeding few microg/l. The most abundant individual compounds belonged to several classes of pharmaceuticals (antimicrobials, analgesics and antiinflammatories, beta-blockers and lipid regulators) and personal care products (fragrances). Due to the rather poor wastewater management practices in WB countries, with less than 5% of all wastewaters being biologically treated, most of the contaminants present in wastewaters reach ambient waters and may represent a significant environmental concern. PMID- 18420256 TI - Pollutant emissions from vehicles with regenerating after-treatment systems in regulatory and real-world driving cycles. AB - Regenerating exhaust after-treatment systems are increasingly employed in passenger cars in order to comply with regulatory emission standards. These systems include pollutant storage units that occasionally have to be regenerated. The regeneration strategy applied, the resultant emission levels and their share of the emission level during normal operation mode are key issues in determining realistic overall emission factors for these cars. In order to investigate these topics, test series with four cars featuring different types of such after treatment systems were carried out. The emission performance in legislative and real-world cycles was monitored as well as at constant speeds. The extra emissions determined during regeneration stages are presented together with the methodology applied to calculate their impact on overall emissions. It can be concluded that exhaust after-treatment systems with storage units cause substantial overall extra emissions during regeneration mode and can appreciably affect the emission factors of cars equipped with such systems, depending on the frequency of regenerations. Considering that the fleet appearance of vehicles equipped with such after-treatment systems will increase due to the evolution of statutory pollutant emission levels, extra emissions originating from regenerations of pollutant storage units consequently need to be taken into account for fleet emission inventories. Accurately quantifying these extra emissions is achieved by either conducting sufficient repetitions of emission measurements with an individual car or by considerably increasing the size of the sample of cars with comparable after-treatment systems. PMID- 18420257 TI - Polymorphonuclear leukocyte phagocytic function increases in plasminogen knockout mice. AB - BACKGROUND: Mice lacking plasminogen (PG-/-) require alternative pathways of fibrinolysis for survival. This may depend on polymorphonuclear leukocytes (PMN) that can clear soluble and insoluble fibrin(ogen) through PG-independent processes. Our objective was to demonstrate that PMNs from PG-/- mice exhibit increased Mac-1 dependent phagocytic activity, which may explain their increased fibrin(ogen)lytic activity compared with wild type (PG+/+) mice. METHODS: Phagocytic activity of PMNs from PG-/- and PG+/+ mice was compared following exposure to Staphylococcus aureus (S. aureus) particles and the expression of Mac 1 was assessed in parallel by flow cytometric analysis. Resistance to phorbol-12 myristate-13-acetate (PMA)-induced cell death was compared between PMNs from the different genotypes. RESULTS: Stimulation of PG-/- PMNs by opsonized S. aureus diluted in PG-/- plasma significantly increased phagocytosis (15-fold) compared with stimulation of PG+/+ PMNs in PG+/+ plasma. Incubation of PG-/- PMNs with PG+/+ plasma (control) or PG-/- plasma supplemented with human PG inhibited this increased phagocytic activity. Mac-1 cell surface density increased 6.2+/-1.0 fold in PG-/- PMNs versus 2.9+/-0.6-fold in PG+/+ PMNs (P < 0.01) indicating that Mac-1 may be associated with increased phagocytic activity. Supporting this, treatment of PG-/- PMNs with an anti-Mac-1 antibody in PG-/- plasma inhibited phagocytic activity. In addition, physiologic PG blocked Mac-1 accessibility at the surface of PMNs. Addition of PMA resulted in 33% death of PMNs from PG-/- mice versus 68% in PG+/+ controls (P < 0.001). CONCLUSIONS: PMNs from PG-/- mice exhibit a Mac-1 dependent increase in phagocytic activity that is suppressed with human PG, an anti-Mac-1 antibody or the plasma from PG+/+ mice. The propensity for PMNs from PG-/- mice to be activated in response to PMA together with their relative resistance to PMA-toxicity may contribute to increased PMN half-life and enhanced fibrin(ogen) clearance in the setting of PG deficiency. PMID- 18420258 TI - Safety of low-dose subcutaneous enoxaparin for the prevention of venous thromboembolism after primary intracerebral haemorrhage. AB - BACKGROUND: The risks and benefits of low molecular weight heparins (LMWH) for the prevention of deep-vein thrombosis (DVT) and pulmonary embolism (PE) after intracerebral haemorrhage (ICH) have not been assessed. The few studies on this subject have revealed conflicting results. METHODS: We retrospectively evaluated whether subcutaneous enoxaparin (20 mg daily) reduced symptomatic venous complications or caused increased 3-month death rate. We included 407 patients who were admitted to a stroke unit and survived the first two days after onset of ICH. There were 232 patients who received anticoagulant treatment for the prevention of DVT and PE and 175 who did not. RESULTS: Despite the fact that the treated patients were in worse clinical condition at the start of the treatment, 3-month death rate was 19% among them compared to 21% among those not receiving anticoagulant therapy. Low-dose subcutaneous enoxaparin (20 mg once daily) induced a significant plasma anti-factor Xa activity 2-3 hours after administration (p=0.018). Haematoma enlargements (33%) occurred in 9% and 7% of the treated and untreated patients, whereas symptomatic venous thromboembolic complications were observed in 3% and 2%, respectively. CONCLUSIONS: We did not observe any increased mortality among ICH patients who survived the first 2 days after the onset of ICH and were thereafter treated with enoxaparin 20 mg daily relative to patients remaining untreated. A randomized trial of the effect of LMWH with a higher dose in prevention of venous thromboembolic complications would be indicated. PMID- 18420259 TI - The activation of apoptosis factor in hindlimb unloading-induced muscle atrophy under normal and low-temperature environmental conditions. AB - In order to identify the apoptosis-induced factors and apoptosis pathway in hindlimb unloading muscle atrophy, the reciprocal relationships between caspase-3 activation and factors related to mitochondria, other organelle pathways, oxidative stress and nitric oxide were investigated. Male Wistar rats were divided into four groups, two groups of hindlimb-unloaded rats were maintained under normal (25 degrees C) and low-temperature (10 degrees C) environmental conditions for a 3-week experimental period, plus two corresponding control groups. Active caspase-3-containing myofibers were observed in the hindlimb unloaded rats in normal and low-temperature environments, but not in the control rats. In these caspase-3-containing fibers, DNA fragmentation, dystrophin breakdown, increased immunolabeling of mu-calpain, decreased cytochrome c, cathepsin-D effusion from the lysosomes and increased lipid peroxidation were observed, while no changes in active caspase-12, eNOS or nNOS immunolabeling were seen. Furthermore, although caspase-3 activation was observed in type-I fibers, caspase-12 labeling was observed in fibers of the hybrid type. These results show that the apoptosis observed in hindlimb unloading-induced muscle atrophy is caused by activation of the caspase cascade via the lysosome pathway. Moreover, the results suggest that caspase-12 does not activate caspase-3 due to differences in the cell differentiation or the apoptosis-inducing stimulation. PMID- 18420260 TI - Regressing amphibian tail as a model for the cadherin/beta-catenin complex disruption and glycosylation alteration during epithelial apoptosis. AB - Epidermis is one of the many tissues that are resorbed during metamorphosis in the regressing tail of amphibian tadpoles. Apoptotic mechanisms play an important role in this process. In this study, loss of intercellular contacts and alterations in plasma membrane glycosylation were observed during apoptosis. The cadherin/beta-catenin complex represents one of the major adhesive systems in multiple epithelial tissues. Here, we analysed the fate of cadherin/beta-catenin complex and alterations of plasma membrane glycoconjugate compositions in apoptotic epithelial cells. Our results showed that the cadherin molecules were cleaved into extracellular and beta-catenin associated cytosolic domains by an intracellular mechanism. However, the extracellular domains were probably removed completely by matrix metalloproteinases. Lectin histochemistry studies suggested that mannose and alpha(2-->6) linked (but not alpha(2-->3) linked) sialic acids were major sugar motifs in plasma membranes of apoptotic tadpole epithelial cells. Although previous studies indicated reduced levels of sialic acid residues during apoptosis, elevated Sambucus nigra agglutinin (SNA) reactivity might be due to the degradation of high molecular weight glycoproteins (probably including cadherin) that masked the SNA-binding residues of the plasma membrane prior to apoptosis. PMID- 18420261 TI - "Self and parent-assessed skin cancer risk factors in school-age children". AB - OBJECTIVE: To evaluate sunburn, sun sensitivity factors and sun protection behavior in school-age children. METHODS: 2002 to 2004 survey of 2942 children in primary schools of Valencia, Spain, and their parents, using a self-administered questionnaire filled by the children with the help of their parents. RESULTS: Having a fair skin (OR: 2.05; 95% CI: 1.38-3.04), light coloured eyes (OR: 1.38; 95% CI: 1.12-1.68), freckles (OR: 1.32; 95% CI:1.12-1.56), and older age (OR: 2.34; 95% CI:1.96-2.80) were associated with occurrence of sunburns. Hair color, gender, use of sunscreens, wearing T-shirts and sunglasses were not. Wearing hats (OR: 0.64; 95% CI: 0.54-0.75) was inversely associated. Parents were significantly more inclined to protect younger and fair-skinned children with sunscreen and T-shirts. CONCLUSIONS: As expected, phenotype is related to sunburns and appears to influence parent's sun protection behaviours. PMID- 18420262 TI - Conventional pluripotency markers are unspecific for bovine embryonic-derived cell-lines. AB - Bovine embryonic stem cells are of potentially big value in transgenic research and studies of lineage commitment and development. Nevertheless, key aspects of the establishment of bovine embryonic stem cells such as the identification of specific pluripotency markers need to be clarified to achieve successful results. Bovine blastocysts were produced in vitro and cultured for 8 days up to the expanded or hatched stage. The trophectoderm, the inner cell mass and its embryonic stem cell-derived lines, all showed a common positive immunocytochemical staining for stage-specific embryonic antigen-4, tumour rejection antigen gp96 and NANOG proteins. The antigenic profile obtained partially agrees with previous data from bovine and other species. Until a validated pluripotent bovine stem cell marker can be identified, it might be advisable to combine the use of epiblast and trophoblast-specific markers to rule out the presence of early committed trophectoderm cells in bovine embryonic stem cell cultures. PMID- 18420263 TI - Microstructural characterization and comparative evaluation of physical, mechanical and biological properties of three ceramics for metal-ceramic restorations. AB - OBJECTIVES: A wide variety of dental ceramics compositions have been introduced in dental clinical practice in order to combine desired aesthetics with superior mechanical performance. The aim of the present study was to investigate the microstructural changes in three dental ceramics after their sintering according to manufacturers' instructions and to comparatively evaluate some of their physical, mechanical and biological properties. METHODS: The analysis of the phases present in each material before and after sintering was performed with scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR) and X-ray diffraction analysis (XRD). The thermal properties of ceramic specimens were evaluated with differential thermal and thermogravimetric analysis (TG-DTA). The mechanical properties evaluated were fracture toughness, Young's modulus and microhardness with the Vickers indentation method. MTT assay was used for cell proliferation assessment. One-way analysis of variance (ANOVA) with Bonferroni multiple comparisons tests was used to determine statistically significant differences (significance level of p<0.05). RESULTS: Results showed a remarkable variation among the three ceramic compositions of leucite content in the starting unheated ceramic powders ranging between 14 and 32 wt.% and in the respective sintered powders ranging between 15 and 41 wt.% The low fusing glass ceramic and the high fusing leucite-based ceramic presented significantly higher fracture toughness (p<0.001) and microhardness and lower modulus of elasticity (p<0.05) compared to the low fusing feldspathic ceramic. The three ceramics were almost equivalent concerning their in vitro biological behavior. SIGNIFICANCE: Variations in crystal structure, distribution and composition are related to differences concerning mechanical properties of dental ceramics. PMID- 18420264 TI - Optical characterization of bisphenol-A-glycidyldimethacrylate triethyleneglycoldimethacrylate (BisGMA/TEGDMA) monomers and copolymer. AB - OBJECTIVE: The fundamental optical characteristics of dental polymers as a function of curing time are exiguously reported in the literature. In this work the refractive index [1] and the extinction coefficient [2] of bisphenol-A glycidyldimethacrylate-triethyleneglycoldimethacrylate (BisGMA/TEGDMA)-based polymers of different volume fractions have been studied as a function of the curing time. A comparison between the measured index of refraction and theoretical calculation based on molar refraction is made. Also the temporal behavior of both the refractive index and extinction coefficient is compared. METHODS: Index of refraction: BisGMA/TEGDMA monomer samples were cured with a commercial light-curing unit by using camphorquinone and DMAEMA as the photoinitiator system. Curing times of 0, 40, 60, 300, 900 and 1800 s were used. The refractive indices were determined with a refractometer at 470 nm wavelength, where the peak absorption of the camphorquinone photoinitiator is located. Extinction coefficient: BisGMA/TEGDMA samples having different lengths were cured and the relative attenuation of light at 470 nm was measured as a function of the curing time (0, 40, 60, 300, 900, 1800 s). RESULTS: The results show that in general the index of refraction increases during the photopolymerization process in a time frame from 0 to 40s and then approaches a steady-state value, which depends on the original composition of the admixture. The detailed refractive index change depends on the volume fractions of the constituent substances. The extinction coefficient decreases in a time frame from 60 to 900 s and reaches a steady-state value thereafter. SIGNIFICANCE: Knowledge of the fundamental optical characteristics of commonly used dental polymers is essential in the development of advanced light-initiated curing methods. PMID- 18420265 TI - Finite element analysis and bond strength of a glass post to intraradicular dentin: comparison between microtensile and push-out tests. AB - OBJECTIVE: This study tested the hypothesis that the stress distribution and bond strength of glass posts to intraradicular dentin is influenced by the mechanical testing methodology. METHODS: Thirty single rooted endodontically treated teeth were prepared for luting of tapered fiber-glass posts (Reforpost, Angelus, Londrina, PR, Brazil) with a conventional adhesive system and resin luting cement (Adper Scotchbond Multi-purpose, Rely X ARC, 3M ESPE, St. Paul, MN, USA). The teeth were randomly divided (n=10 per group) into micro-push-out (Mpo), hourglass (Mh) and rectangular stick-shaped (Ms) microtensile testing groups before sectioning each root into five 1-mm-thick specimens. During specimen preparation for microTBS testing 46/50 stick and 4/50 hourglass specimens prematurely failed; therefore, the Ms group could not be included in the mechanical testing. The remaining specimens were tested at 0.5 mm/min until bond failure. Stress distribution within each specimen type for the three mechanical test methods was analyzed by finite element analysis (FEA). Qualitative analyses were carried out through Von Mises, XY and Sy criterion. RESULTS: Mpo and Mh had a mean microTBS of 11.89+/-6.55 and 14.98+/-12.72 MPa, respectively, which was not significantly different (p=0.1311). The push-out test demonstrated a more homogenous stress distribution by FEA and less variability in mechanical testing. SIGNIFICANCE: Therefore, the recommended testing method for determining the bond strength of glass posts to intraradicular dentin is by Mpo. PMID- 18420266 TI - Detection of environmental mutagens using the FACIM assay. AB - A genetically engineered diploid yeast strain named yJC2, was specifically developed for environmental mutagen detection and characterization of induced mutations. This strain contains one copy of the human TP53 tumour suppressor gene coding sequence which is used as a molecular target for mutagens and two copies of the ADE2 reporter gene allowing accurate measurement of the TP53 transcriptional activity. The strain sensitivity to mutagens was evaluated by exposing cells to UVC, 4-nitroquinoline (NQO) or to an organic extract of sediment from the Seine estuary. For all studied mutagens, a significant and dose dependent increase of mutant frequency was observed. The present assay named FACIM II (Functional Analysis of Chemical-Induced TP53 Mutations) is more convenient than the FACIM I and more inducible than the SOS Chromotest to detect direct-acting mutagens in the environment. PMID- 18420267 TI - Enzymatically crosslinked collagen-mimetic dendrimers that promote integrin targeted cell adhesion. AB - Collagen is made up of a diverse family of the extracellular matrices, most of which are generally found crosslinked in vivo. To more closely mimic the biological function of collagen, this work focuses on establishing a molecular strategy to engineer a functional biomimetic collagen that exhibits stable collagen-like triple-helical conformation with cell-binding activity, in addition to an enzyme-mediated crosslinking by tissue transglutaminase (tTGase). A novel sequence spanning residues 2800-2807 of human fibrillin-1 (EDGFFKI) was first identified as an amine donor substrate for tTGase, using a previously characterized APQQEA derived from human osteonectin as an amine acceptor probe. Subsequently, collagen-mimetic peptides (CMPs) supplemented with a cell-binding sequence (GFOGER) and the identified EDGFFKI and APQQEA substrate sequences were conjugated onto a generation 2 poly(amidoamine) dendrimer, resulting in a crosslinkable collagen-mimetic dendrimer, denoted as CMD-K and CMD-Q, respectively. Both CMD-K and CMD-Q exhibited enhanced triple-helical stability and supported cell adhesion in an integrin-specific manner. Finally, tTGase mediated crosslinking between CMD-K and CMD-Q resulted in a supramolecular structure that exhibited stable collagen-like triple-helical conformation and improved cellular recognition. The results show that the triple-helical structure is important in preserving the GFOGER cell-binding site while the tTGase-mediated protein crosslinking may also be crucial for the recognition by cell surface integrin receptors. PMID- 18420268 TI - Morphological and electrical properties of human trophoblast choriocarcinoma, BeWo cells. AB - The syncytiotrophoblast of the human placenta arises from fusion of stem cells called cytotrophoblasts. The molecular mechanisms associated with cell fusion and syncytiation of cytotrophoblastic cells remain largely unknown. In the present study, we investigated the morphological and electrical properties of BeWo cells, a human choriocarcinoma-derived trophoblast cell model, with several features of the human cytotrophoblast. Cultured cells tended to cluster, but only fused into small, multinucleated syncytia in the presence of cAMP (72 h). The morphological features of both the actin and microtubular cytoskeletons indicated that within 72 h of constant exposure to cAMP, intracellular cortical actin cytoskeleton disappeared, which was the most prominent inducing factor of multi-nucleation. The presence of the cation channel protein, polycystin-2 (PC2), a TRP-type cation channel, associated with placental ion transport in term human syncytiotrophoblast, co-localised with acetylated tubulin in midbodies, but was found non-functional under any conditions. Different electrical phenotypes were observed among control BeWo cells, where only 26% (8 of 31 cells) displayed a voltage-dependent outwardly rectifying conductance. Most quiescent BeWo cells had, however, a low, slightly outwardly rectifying basal whole cell conductance. Acute exposure to intracellular cAMP (<15 min) increased the whole cell conductance by 122%, from 0.72 nS/cell to 1.60 nS/cell, and eliminated the voltage-regulated conductance. The encompassed evidence indicates that the early events in BeWo cell fusion and syncytiation occur by cAMP-associated changes in ionic conductance but not morphological changes associated to chronic exposure to the second messenger. This suggests a tight regulation, and important contribution of cation conductances in cytotrophoblastic cells prior to syncytiation. PMID- 18420269 TI - STIM1 converts TRPC1 from a receptor-operated to a store-operated channel: moving TRPC1 in and out of lipid rafts. AB - While the role of members from the TRPC family of channels as receptor-operated channels (ROC) is well established and supported by numerous studies, the role of this family of channels as store-operated channels (SOC) has been the focus of a heated controversy over the last few years. In the present study, we have explored the modulation of STIM1 on human TRPC1 channel. We show that the association of STIM1 to TRPC1 favors the insertion of TRPC1 into lipid rafts, where TRPC1 functions as a SOC. In the absence of STIM1, TRPC1 associates to other members from the TRPC family of channels to form ROCs. A novel TIRFM-FRET method illustrates the relevance of the dynamic association between STIM1 and TRPC1 for the activation of SOC and the lipid raft localization of the STIM1 TRPC1 complex. This study provides new evidence about the dual activity of TRPC1 (forming ROC or SOC) and the partners needed to determine TRPC1 functional fate. It highlights also the role of plasma membrane microdomains and ER-PM junctions in modulating TRPC1 channel function and its association to STIM1. PMID- 18420270 TI - Twin pregnancy in a patient of chronic myeloid leukemia on imatinib therapy. AB - Imatinib is a tyrosine kinase inhibitor and is now used regularly in chronic myeloid leukaemia therapy in chronic phase with great success. This drug due its very nature of action is suspected to be teratogenic hence the patients are counseled not to get pregnant while on this drug. However in world literature few normal pregnancies have been reported in patients on Imatinib therapy, though no twin pregnancy has been reported on this medication. We report here the birth of normal mono-ovular mono-chorionic twin while the patient is on imatinib during conception and early pregnancy for chronic myeloid leukaemia. PMID- 18420271 TI - Characterization of macrophages from the bony fish gilthead seabream using an antibody against the macrophage colony-stimulating factor receptor. AB - Two major professional phagocyte populations have been described in fish, namely granulocytes and monocytes/macrophages. Although the distribution and localization of macrophages have been documented in several teleost species using mainly light and/or electron microscopy, the lack of appropriate markers for these cells has hampered our in-depth knowledge of their biology. We report here the generation of a monospecific rabbit polyclonal antibody against the gilthead seabream macrophage colony-stimulating factor receptor (Mcsfr), which is an excellent marker of macrophages in mammals and the zebrafish. The anti-Mcsfr has been found to be very useful in immunohistochemistry (IHC) to specifically immunostain the purified macrophages (adherent cells) obtained from the head kidney as well as different cell populations in paraffin-embedded organs, including the head-kidney, spleen, thymus, gills and liver. Unexpectedly, however, no Mcsfr immunoreactive (Mcsfr(+)) cells were observed in the brain and intestine of the gilthead seabream. We also show that the distribution of Mcsfr(+) cells in the head-kidney and the spleen is unaltered following infection with the fish pathogenic bacterium Vibrio anguillarum and that the Il1b-producing cells in these two organs after infection are exclusively acidophilic granulocytes. Finally, as the epitope recognized by the anti-Mcsfr is well conserved, we illustrate the potential usefulness of this antibody in other teleost species, such as the European seabass. PMID- 18420272 TI - Contribution of the apparent diffusion coefficient in perilesional edema for the assessment of brain tumors. AB - OBJECTIVES: Diffusion-weighted MRI is sensitive to molecular motion and has been applied to the diagnosis of stroke. Our intention was to investigate its usefulness in patients with brain tumor and, in particular, in the perilesional edema. METHODS: We performed MRI of the brain, including diffusion-weighted imaging and mapping of the apparent diffusion coefficient (ADC), in 16 patients with brain tumors (glioblastomas, low-grade gliomas and metastases). ADC values were determined by the use of regions of interest positioned in areas of high signal intensities as seen on T2-weighted images and ADC maps. Measurements were taken in the tumor itself, in the area of perilesional edema and in the healthy contralateral brain. RESULTS: ADC mapping showed higher values of peritumoral edema in patients with glioblastoma (1.75 x 10(-3)mm(2)/s) and metastatic lesions (1.61 x 10(-3)mm(2)/s) compared with those who had low-grade glioma (1.40 x10( 3)mm(2)/s). The higher ADC values in the peritumoral zone were associated with lower ADC values in the tumor itself. CONCLUSIONS: The higher ADC values in the more malignant tumors probably reflect vasogenic edema, thereby allowing their differentiation from other lesions. PMID- 18420273 TI - Occlusion of the artery of percheron: clinical and neuroimaging correlation. PMID- 18420274 TI - [A survey of the management of unruptured intracranial aneurysms as practised by French neuroradiological and neurosurgical teams]. AB - BACKGROUND: The lack of consensus in the management of unruptured intracranial aneurysms (UIA) has resulted in a variety of different clinical practices. The aim of this study is to analyze these different practices. METHODS: A questionnaire concerning the management of UIA was mailed out to French neurosurgeons (NS) and neuroradiologists (NR). Eighteen responses from 17 teams of NS and 23 responses from 19 teams of NR were included in our analysis. RESULTS: In making a therapeutic decision, about three-quarters of both NR and NS take into account the age of the patient and all of our responders except one consider the aneurysm's morphology, especially its size and neck structure. Pinpointing the location of the aneurysm is an important factor for 61% of NR and 40% of NS. Information concerning the risk of aneurysm rupture and the risks of treatment is routinely given to the patient orally and, sometimes, in writing. The follow-up of UIA treated by NR usually consists of one X-ray angiography and several MR angiographic (MRA) films taken over a period of at least five years and, sometimes, for the rest of the patient's life (22%). The follow-up after surgical treatment mainly comprises X-ray angiography for a limited period of time-usually from five to ten years. The follow-up of untreated aneurysms is usually by either MRA or angioCT. For most NR, the duration of follow-up is long and, sometimes, unlimited. For NS, the duration is more difficult to pinpoint: the response was indeterminate in 28 and 33% gave no response at all. If the first screening tests negative, 44% of NS and 61% of NR propose a repeat screening. CONCLUSION: Given the differences in the management of UIA as revealed by this survey, a multidisciplinary approach that combines the various clinical practices may be the best way forward. PMID- 18420275 TI - Mycoplasma pneumoniae in Korean children: the epidemiology of pneumonia over an 18-year period. AB - OBJECTIVES: Mycoplasma pneumoniae (MP) is a major cause of community-acquired pneumonia in children. The aim of this study was to investigate the long-term epidemiology of MP pneumonia in Korean children. METHODS: A retrospective analysis of a database of 2405 patients with pneumonia at the Seoul National University Children's Hospital between 1986 and 2004 was performed. Serologic diagnosis for MP infection was made based on a 4-fold rise or single titers >/=1:640, which were measured by an indirect agglutination test. RESULTS: MP pneumonia was diagnosed in 568 patients over 18 years. The mean age was 5.7 years. Children younger than 5 years of age accounted for 44% of the cases. Six outbreaks were observed at intervals of 3-4 years. The earlier epidemics up until 1996 peaked in the summer, while the later epidemics peaked in the fall or early winter. Children <5 years old were more commonly affected during large epidemics compared to endemic periods. The geometric mean antibody titers were maintained >/=1:320 up to 7 months after the onset of illness. CONCLUSIONS: The results of this study revealed community outbreaks of MP pneumonia at 3-4-year intervals among Korean children. A significant proportion of young Korean children were affected by MP pneumonia, especially during large epidemics. PMID- 18420276 TI - High serum levels of neopterin in patients with Crimean-Congo hemorrhagic fever and its relation with mortality. AB - OBJECTIVE: Neopterin is generated and released in increased amounts by macrophages upon activation by interferon-gamma during cellular immune response. In this study, we aimed to investigate serum neopterin levels in patients with Crimean-Congo hemorrhagic fever (CCHF) and its clinical significance as a predictor factor of mortality. METHODS: Neopterin concentrations on the first day of hospitalization were measured in serum samples from 51 CCHF patients. Serum neopterin levels and other clinical-laboratory parameters for fatal and nonfatal CCHF patients were compared. RESULTS: Serum neopterin levels (73.22+/-54.30 nmol/L) were highly elevated in all CCHF patients (p<0.0001) with higher levels in fatal group (153.66+/-81.34 nmol/L, p=0.0001) compared to nonfatal disease (55.99+/-24.09 nmol/L). In univariate analysis, the level of neopterin on the first day of hospitalization, bleeding, platelet count, aspartate transferase and lactate dehydrogenase were associated with mortality. In multivariate analysis, only the serum level of neopterin was associated with mortality. As a mortality risk factor, area under the curve was 0.939 (p=0.0001, 95% confidence interval: 0.85-1.00). CONCLUSIONS: In this first study of serum neopterin levels for CCHF, elevated serum neopterin level showing strong activation of monocytes/macrophages was a risk factor for CCHF. PMID- 18420277 TI - Cyclooxygenase and prostaglandin synthases in atherosclerosis: recent insights and future perspectives. AB - Cyclooxygenase (COX) is the key enzyme in the conversion of arachidonic acid to prostanoids, lipid mediators involved in several physiological and pathological processes. Two COX isoenzymes have been characterized, COX-1 and COX-2, that differ in terms of regulatory mechanisms of expression, tissue distribution, substrate specificity, and preferential coupling to upstream and downstream enzymes. Both isoforms play fundamental roles in atherothrombosis; however, whereas the function of COX-1 in this setting is well established, the role of COX-2 remains unclear. Indeed, the intracellular pathways regulating COX-2 induction appear numerous and complicated, varying between cell types and cellular stimulus. In recent years a long series of studies has been performed with the aim of clarifying the role of COX-2 in atherothrombosis, with the major finding that the COX-2 expression pattern in arterial vessels may be associated with either protective or plaque-destabilizing phenotypes according to the downstream synthase that couples with COX-2. In this review we summarize the role of COX-2 as well as the different downstream synthases in atherosclerosis and atherothrombosis. Finally, we briefly review the controversial vascular effects on prostanoid inhibition by COX-2 inhibitors. PMID- 18420278 TI - Assessing QT prolongation in conscious dogs: validation of a beat-to-beat method. AB - A model of sling-trained, conscious mongrel dogs instrumented with telemetric arterial pressure transmitters and ECG leads was validated for assessment of the QT-RR interval relationship at clinically used free and total plasma concentrations of positive and negative standards with known outcomes. The beat to-beat technique for assessing the dynamic boundaries of the individual cardiac cycles was compared to the same data with typically used averaging techniques and corrections applied. Positive standards E-4031, cisapride, terodiline, and terfenadine showed increased sensitivity toward detection at clinically relevant levels when an outlier analysis of beats beyond the normal autonomic boundary is applied. Since methods to correct the QT interval for heart rate are often confounded with changes in autonomic state, a validation of the changes with reflex tachycardia induced by vasodilatation after nitroprusside and reflex bradycardia induced by sudden vasoconstriction with phenylephrine where shown to be differentiated from direct effects of repolarization with E-4031. These changes were also demonstrated to be identical to effects observed in humans after standing or challenged with a similar dose of phenylephrine. The conscious dog is also a sensitive model for studying the arrhythmia liability induced by beat-to-beat changes in cardiac ECG restitution (the relationship between QT and TQ intervals) and hysteresis. However, some caveats based on observations may need to be considered due to inherent differences in QT intervals and sinus arrhythmia between canines and humans. PMID- 18420279 TI - Taming the PI3K team to hold inflammation and cancer at bay. AB - Recent progress in understanding the molecular mechanisms of receptor signal transduction is continuously highlighting new unforeseen potential drug targets for yet unmet therapeutic needs. While the large number of different cell surface receptors challenge the concept of antagonists development, the finding of signal transduction platforms common to multiple receptor families has boosted the development of new therapeutic approaches. The identification of the role of phosphoinositide 3-kinase family members downstream receptors as directors of multiple cellular responses ranging from cell proliferation and survival to immunity and cardiovascular control, is an example of successful drug target validation studies. This review will focus on these findings and on the ongoing efforts to tame this family of enzymes to beat inflammation and cancer. PMID- 18420280 TI - A glue-based, screw-free method for implantation of intra-cranial electrodes in young mice. AB - Intra-cranial electroencephalographic recordings are increasingly employed in mice because of the availability of genetically manipulated mouse models. Currently, dental acrylic and anchoring screws are used to cement implanted electrodes. This technique works well for adult animals but often encounters difficulty when employed in young mice because their skulls are not strong enough to bear the anchoring screws. Here we describe a novel method favorable for implantation of intra-cranial electrodes in mice as young as postnatal 18 days and suitable for long-term intra-cranial electroencephalographic recordings. Our approach is to construct a multi-electrode assembly according to the desired stereotaxic coordinates of intra-cranial recordings and to secure the implanted electrode assembly to the skull via glue rather than dental acrylic/anchoring screws. The surgical operation for such electrode implantation is relatively quick and rarely associated with complications such as infection, bleeding, neurological deficits, spontaneous seizures or behavioral disturbances. The implanted electrodes are stable, allowing repeated monitoring for several months. Data obtained by simultaneous intra-hippocampal and intra-cortical recordings indicate that our method is suitable for the examination of behaviorally related electroencephalographic activities and experimentally induced seizures. Technical aspects of our methods are discussed, and the procedures for constructing the electrode assembly are presented in detail. PMID- 18420281 TI - Artificial dural sealant that allows multiple penetrations of implantable brain probes. AB - This study reports extensive characterization of the silicone gel (3-4680, Dow Corning, Midland, MI), for potential use as an artificial dural sealant in long term electrophysiological experiments in neurophysiology. Dural sealants are important to preserve the integrity of the intracranial space after a craniotomy and in prolonging the lifetime and functionality of implanted brain probes. In this study, we report results of our tests on a commercially available silicone gel with unique properties that make it an ideal dural substitute. The substitute is transparent, elastic, easy to apply, and has re-sealing capabilities, which makes it desirable for applications where multiple penetrations by the brain probe is desirable over an extended period of time. Cytotoxicity tests (for up to 10 days) with fibroblasts and in vivo tests (for 12 weeks) show that the gel is non-toxic and does not produce any significant neuronal degeneration when applied to the rodent cortex even after 12 weeks. In vivo humidity testing showed no sign of CSF leakage for up to 6 weeks. The gel also allows silicon microprobes to penetrate with forces less than 0.5 mN, and a 200-microm diameter stainless steel microprobe with a blunt tip to penetrate with a force less than 2.5 mN. The force dependency on the velocity of penetration and thickness of the gel was also quantified and empirically modeled. The above results demonstrate that the silicone gel (3-4680) can be a viable dural substitute in long-term electrophysiology of the brain. PMID- 18420283 TI - HPA axis exhaustion and regulatory T cell accumulation in patients with a functional somatic syndrome: recent view on the problem of Gulf War veterans. AB - The authors proceeded from the assumption that physical and mental symptoms of functional somatic syndromes (including those observed in Gulf War veterans) are based on both underactivity of hypothalamic-pituitary-adrenal (HPA) axis and excessive accumulation of regulatory T cells (Tregs). Permanent psychogenic stress coupled with high antigen loading leads to gradual depletion of HPA axis, which is manifested by the reduction of stress-induced cortisol response. Under stress hormone deficiency, Tregs begin to play a principal role in anti inflammatory mechanisms and each new pro-inflammatory stimulus increases their number. Superfluous accumulation of active Tregs results in malfunction of Th1 cells in the brain that leads to the appearance of neurodegeneration foci, which seems to be an anatomic substance for various cognitive and psychological symptoms. New approaches to the treatment of such conditions are also discussed. PMID- 18420282 TI - Antibodies to two ZP3 B cell epitopes affect zona pellucida assembly. AB - Mouse zona pellucida (ZP) proteins are synthesized in developing oocytes and assembled into ZP after their secretion. This study has investigated whether anti ZP3 antibodies affect ZP assembly. Peptides CP2 and CP3 were used to elicit antibodies to two ZP3 B cell epitopes, ZP3 (335-342) and ZP3 (171-180). Ovulated eggs from mice immunized with a mixture of CP2/CP3 showed an abnormal ZP; importantly, the ZP completely dissolved both in vitro and in vivo 12h after ovulation. Although CP3 immunization resulted also in abnormal ZP, the ZP did not dissociate. Binding of antibodies to the ZP prior to oocyte maturation was requisite, as in vitro incubation of ovulated eggs in combination with the two antibodies failed to induce ZP dissolution. Electron microscopic observation further demonstrated a significant abnormality in ZP structure in CP2/CP3 immunized mice, especially in mature follicles, suggesting that B cell epitopes may be involved in ZP assembly. Though antibody elicited by CP2 has been shown to inhibit fertilization, we now show that antibody induced by CP3 had no effect on fertility. However, immunization with CP3/CP2 resulted in a significantly lower fertility rate than CP2 alone. This suggests that infertility in these mice may be due to an unstable ZP structure. Our model provides a useful tool to study ZP assembly and its structure beyond molecular biology method. PMID- 18420284 TI - A systematic review of the incidence of sensorineural hearing loss in neonates exposed to Herpes simplex virus (HSV). AB - OBJECTIVE: To develop evidence-based guidelines for appropriate audiological monitoring of children born following exposure to or infection with Herpes simplex virus (HSV) for development of immediate or delayed-onset of sensorineural hearing loss (SNHL). DATA SOURCES: A Medline search of the 1966 July 2007 database was supplemented by search of the additional database Embase (1980-July 2007). Manual search was conducted of references of identified papers and book chapters. STUDY SELECTION: Articles were sought that were longitudinal in design, to include an inception cohort of children infected with (or exposed to) HSV who were entered at a similar point at birth and followed over time with serial audiometry to identify hearing loss if it developed. DATA EXTRACTION: Patient information and audiometric data extraction from relevant articles was performed independently by all three researchers. Discrepancies were resolved by mutual consensus. DATA SYNTHESIS: Data was analyzed using descriptive statistics. RESULTS: Three papers reported five children with SNHL following apparent disseminated HSV-2 infections in which other obvious clinical sequelae of HSV infection and co-morbid conditions were present. Audiometric information is lacking regarding onset and progression. There are no reports of delayed-onset SNHL following perinatal or asymptomatic HSV infection. CONCLUSIONS: The development of SNHL in children with exposure to HSV occurs rarely. Routine serological screening for HSV infection in otherwise healthy neonates newly diagnosed with SNHL is unjustified. There is insufficient data to define the incidence and natural history of SNHL in children with HSV infections. Carefully designed and conducted studies are needed to address this issue. PMID- 18420285 TI - Pressure-inactivated yellow fever 17DD virus: implications for vaccine development. AB - The successful Yellow Fever (YF) vaccine consists of the live attenuated 17D-204 or 17DD viruses. Despite its excellent record of efficacy and safety, serious adverse events have been recorded and influenced extensive vaccination in endemic areas. Therefore, alternative strategies should be considered, which may include inactivated whole virus. High hydrostatic pressure has been described as a method for viral inactivation and vaccine development. The present study evaluated whether high hydrostatic pressure would inactivate the YF 17DD virus. YF 17DD virus was grown in Vero cells in roller bottle cultures and subjected to 310MPa for 3h at 4 degrees C. This treatment abolished YF infectivity and eliminated the ability of the virus to cause disease in mice. Pressure-inactivated virus elicited low level of neutralizing antibody titers although exhibited complete protection against an otherwise lethal challenge with 17DD virus in the murine model. The data warrant further development of pressure-inactivated vaccine against YF. PMID- 18420286 TI - Tactile intensity and population codes. AB - An important question in neuroscience is how different aspects of a stimulus are encoded at different stages of neural processing. In this review, I discuss studies investigating the peripheral neural code for perceived intensity in touch. One of the recurrent themes in this line of research is that information about stimulus intensity is encoded in the activity of populations of neurons. Not only is information integrated across afferents of a given type, but information is also combined across submodalities to yield a unified percept of stimulus intensity. The convergence of information stemming from multiple submodalities is particularly interesting in light of the fact that these are generally thought to be parallel sensory channels with distinct sensory functions and little cross-channel interactions. I discuss implications of a recently proposed model of intensity coding for psychophysical functions and for the coding of intensity in cortex. I also briefly review the peripheral coding of intensity in other sensory modalities. PMID- 18420287 TI - Differences in nociceptive behavioral performance between C57BL/6J, 129S6/SvEv, B6 129 F1 and NMRI mice. AB - The interpretation of knockout and transgenic mouse studies in pain research critically depends on detailed knowledge of the performance profile of the background strains. Pain-related behavior was compared between four relevant mouse strains (C57BL/6J, 129S6/SvEv, B6 129 F1 and NMRI mice of both sexes) using an extended test battery that included an unusual variety of assays for thermal and mechanical acute nociception, and inflammatory and neuropathic pain. Strain- and gender-dependent differences were demonstrated in many of these nociceptive assays. Particularly, C57BL and 129 mice, which serve as the default genetic backgrounds for experiments in genetically altered mice, display quite different patterns of nociceptive performance. Compared to C57BL/6J mice, 129S6/SvEv animals are less sensitive to inflammatory pain conditions (thermal sensitivity after carrageenan subplantar injection; flinch behavior after formalin injection), while the opposite is observed in the neuropathic pain condition and the visceral pain model. These data may be of special interest for genetic studies, where issues related to the background phenotype may confound their interpretation. PMID- 18420288 TI - Synergistic effects of selegiline and donepezil on cognitive impairment induced by amyloid beta (25-35). AB - Selegiline, an irreversible inhibitor of monoamine oxidase B used in the treatment of Parkinson's disease, has been demonstrated to have a potential cognition-improving effect in patients with Alzheimer's disease (AD) undergoing treatment with an acetylcholinesterase inhibitor donepezil. To confirm such clinical events, we investigated whether co-administration of donepezil with selegiline had a synergistic cognition-improving effect in an animal model of AD. Intracerebroventricular injection of amyloid beta protein fragment 25-35 [Abeta(25-35)] induced impairment of learning and memory in a Y-maze, novel object recognition and contextual fear conditioning tests. Either donepezil or selegiline alone improved the cognitive impairments in the Y-maze and conditioned fear learning tasks in Abeta(25-35)-injected mice, whereas donepezil, but not selegiline, failed to improve the impairment in a novel object recognition task. Co-administration of donepezil with selegiline, at doses that do not exert efficacy individually, significantly improved the deficits in all three tests, indicating a synergistic cognition-improving effect. These alleviating effects were antagonized by pretreatment with a muscarinic receptor antagonist scopolamine and a dopamine receptor antagonist haloperidol. These results suggest that selegiline potentiates the effect of donepezil on the cognitive impairment, and that the synergistic effect may be mediated through both the cholinergic and dopaminergic systems. PMID- 18420289 TI - Combined use of Daphnia magna in situ bioassays, biomarkers and biological indices to diagnose and identify environmental pressures on invertebrate communities in two Mediterranean urbanized and industrialized rivers (NE Spain). AB - Environmental factors affecting aquatic invertebrate communities were assessed using Daphnia magna in situ bioassays and biological indices based on community assemblages of benthic macroinvertebrates. Investigations were carried out in two heavily industrialized and urbanized river basins from the NE of Spain (Llobregat and Besos). Measures of energy consumption (i.e. algal grazing), and of specific biochemical responses (biomarkers) were conducted on individuals transplanted upstream and downstream from effluent discharges of sewage treatment plants. In both rivers there was a clear deterioration of the ecological water quality parameters and benthic communities towards downstream reaches. In all but one of the 19 locations studied, transplanted organisms were affected in at least one of the five measured responses. In three of them, significant effects were detected in most of the traits considered. Principal Component and Partial Least Square Projections to Latent Structures regression analyses indicated that the measured responses in D. magna in situ bioassays and those of macroinvertebrate assemblages were affected by distinct environmental factors. From up to 20 environmental variables considered, seven of them including habitat degradation, suspended solids, nitrogenous and conductivity related parameters affected macroinvertebrate assemblages. On the other hand, levels of organophosphorus compounds and polycyclic aromatic hydrocarbons were high enough to trigger the responses of D. magna in situ bioassays. These results emphasize the importance of combining biological indices with biomarkers and more generalized and ecologically relevant (grazing) in situ responses to identify ecological effects of effluent discharges from sewage treatment plants in surface waters. PMID- 18420290 TI - A systematic review of the effectiveness of Tai Chi on fall reduction among the elderly. AB - Falls among the elderly is a major public health concern. There has been recent extensive research on the effects of Tai Chi in fall prevention among the elderly. As such, we undertook a systematic review to look for evidence on the effect of this intervention. There were seven randomized controlled trials, which met our objective and inclusion criteria. Our review has shown that Tai Chi has the potential to reduce falls or risk of falls among the elderly, provided that they are relatively young and non-frail. Further review is needed to look into the non-English studies, which assess the effectiveness of Tai Chi on fall reduction. PMID- 18420291 TI - International programs and veterinary public health in the Americas--success, challenges, and possibilities. AB - The veterinary public health (VPH) program at the Pan American Health Organization (PAHO) began in 1949 when an arrangement with the newly founded World Health Organization made PAHO its Regional Office for the Americas to serve as the specialized health agency both for the Organization of American States and the United Nations. It started as a Section of Veterinary Medicine to help eradicate rabies on both sides of the US-Mexico border, and PAHO grew to be the biggest VPH program in the world. By providing a political and technical base, PAHO assisted its member states to organize and develop their national VPH programs and activities, and it provides technical cooperation and works with their national counterparts to solve national and local problems. In the 1980s and 1990s, PAHO concentrated that cooperation on several, specific needs: the elimination of dog-transmitted human rabies, hemispheric eradication of foot-and mouth disease (FMD), regional action planning for food safety, control/eradication of bovine tuberculosis and brucellosis, and surveillance and prevention of emerging zoonoses and food-borne diseases. The Pan American centers developed a number of diagnostic antigens and a continental system for the surveillance of FMD and vesicular diseases, using geographic quadrant technology to augment sensitivity, analyze data, and make decisions. Another visible accomplishment is the elimination of hydatidosis in the endemic countries and regions of the southern cone. In addition, the VPH program of PAHO pioneered the mobilization of the private sector to participate in official programs. Nevertheless, privatization of animal and human health services has had a negative effect on human resources and infrastructure by weakening essential epidemiological functions in some countries. Today, there is a need for closer coordination between veterinary medicine and medical services. Practically all potential bioterrorism agents are zoonoses, and it is cost-effective to control them at the veterinary level, providing the first line of defense. The opportunities for VPH are boundless, but the challenge is to be able to apply the plethora of available research results and knowledge. What we will need is a new breed of veterinarians who will lead and provide us with a vision, like those we honored in 2005 at the Schwabe Symposium Honoring the Lifetime Achievements of Dr. James H. Steele: veterinarians in public health who will be in the forefront of policy setting, decision-making, and allocation of resources, and veterinarians who will articulate and provide a strategic direction to our unique professional skills. PMID- 18420292 TI - Exploring the foundations of population health and preventive medicine as essential elements for veterinary education. AB - The evolution of preventive medicine and public health training in professional veterinary medicine curricula is documented. Most veterinary colleges in the US began with a single course in meat hygiene or public health, with a focus on food hygiene issues. These courses laid the foundation for modern veterinary preventive medicine and public health training for veterinary students. Most graduates of veterinary colleges today have extensive training in population health, preventive medicine, and zoonotic diseases. PMID- 18420293 TI - Start spreading the news: a structural estimate of the effects of New York hospital report cards. AB - Research on the effects of publicly reported hospital quality report cards on patient market shares is mixed. Higher-ranking hospitals do not consistently experience increases in market share. We argue that this may be because the report cards do not always convey "news" about quality; in some cases the rankings conform with prior beliefs about quality. We develop a structural model of the "news" in report cards and estimate the model using data from New York State in 1989-1991. We show hospitals with negative news in the original 1990 report cards experienced a decrease in market share, but that a misspecified model might continue to find no report card effect. PMID- 18420294 TI - GP supply and obesity. AB - We investigate the relationship between area general practitioner (GP) supply and individual body mass index (BMI) in England. Individual level BMI is regressed against area whole time equivalent GPs per 1000 population plus a large number of individual and area level covariates. We use instrumental variables (area house prices and age weighted capitation) to allow for the endogeneity of GP supply. We find that that a 10% increase in GP supply is associated with a mean reduction in BMI of around 1kg/m(2) (around 4% of mean BMI). The results suggest that reduced list sizes per GP can improve the management of obesity. PMID- 18420295 TI - A versatile plasmid biosensor useful to identify quorum sensing LuxR-family orphans in bacterial strains. AB - Eight luxI-family gene promoters (luxI, cviI, ahlI, rhlI, cepI, phzI, traI and ppuI) were cloned in tandem, upstream a promoterless lacZ gene in a promoter probe vector yielding pMULTIAHLPROM. This unique construct is useful in determining whether a bacterial strain not producing N-acyl homoserine lactone signal molecules (AHLs) possesses orphan LuxR type proteins able to respond to AHLs and activate transcription from quorum sensing target genes. Using pMULTIAHLPROM, it was demonstrated that Enterobacter aerogenes possibly contains a LuxR-family orphan able to activate luxI-family promoters independently from AHLs. PMID- 18420296 TI - In vivo alanine/electron spin resonance (ESR) dosimetry in radiotherapy of prostate cancer: a feasibility study. AB - PURPOSE: We have developed a device to evaluate the potential of alanine/electron spin resonance (ESR) dosimetry for quality assurance in 3D conformal radiotherapy for prostate cancer. It consists of a rectal balloon carrying eight alanine dosimeter probes and two metal markers to document the exact position of the balloon. We measured the effects of an air-filled rectal balloon on the dose at the rectal wall and compared these results with the applied dose distribution of the treatment planning system. MATERIALS AND METHODS: During 10 fractions with 2.0 Gy per fraction, the accumulated doses were measured in 3 patients. The results of the ESR measurements were compared to the applied doses. RESULTS: It was possible to insert the device without clinical complications and without additional rectal discomfort for the patients. The measurements of the dose accumulated at the anterior and the posterior rectal wall agreed with the applied dose within a mean deviation of 1.5% (overestimation of the dose) and 3.5% (underestimation of the dose), respectively. However, clinically significant differences between applied and measured rectal doses were seen in a patient with a hip prosthesis. In this case, the dose at the anterior rectal wall was overestimated by the TPS by about 11% and the dose at the posterior rectal wall was underestimated by approximately 7%. CONCLUSION: The method presented in this study is useful for quality control of irradiations in vivo. PMID- 18420297 TI - Synergistic effects of sequential treatment with methyl jasmonate, salicylic acid and yeast extract on benzophenanthridine alkaloid accumulation and protein expression in Eschscholtzia californica suspension cultures. AB - To develop an optimal bioprocess for secondary metabolite production and explain the bioprocess at the molecular level, we examine the synergistic effects of sequential treatment with methyl jasmonate (MJ), salicylic acid (SA) and yeast extract (YE) on benzophenanthridine alkaloid accumulation and protein expression in Eschscholtzia californica suspension cultures. Serial treatment of MJ, SA and YE at 24h intervals enhanced the accumulation of dihydrosanguinarine (2.5 times) and sanguinarine (5.5 times). This sequential treatment using different signal elicitors was more effective than single elicitor or simultaneous treatment of the elicitors; it induced benzophenanthridine alkaloid accumulation to 917.7+/ 42.0mg/L. Also, (S)-methylcoclaurine-3'-hydroxylase (CYP80B1) and 3'-hydroxy-(S) N-methylcoclaurine-4'-O-methyltransferase (4'OMT) expressions among enzymes in sanguinarine biosynthetic pathway explained the synergistic effects by sequential treatment of the elicitors. The sequential treatment strategy using elicitors related to different signal transduction pathways can be used to design better processes to increase accumulation of secondary metabolites in plant cell culture. Analysis of protein expression provides the detailed information about metabolite accumulation through the correlated results. PMID- 18420298 TI - A novel self-deleting retroviral vector carrying an additional sequence recognized by the viral integrase (IN). AB - During retroviral integration, the viral integrase recognizes the attachment (att) sequence (formed by juxtaposition of two LTRs ends) as the substrate of integration. We have developed a self-deleting Avian Leukosis and Sarcoma Viruses (ALSVs)-based retroviral vector carrying an additional copy of the att sequence, between neo and puro genes. We observed that: (i) the resulting NP3Catt vector was produced at neo and puro titers respectively smaller and higher than that of the parental vector devoid of the att sequence; (ii) 61% of NP3Catt proviruses were flanked by LTRs; most of them were deleted of internal sequences, probably during the reverse transcription step; (iii) 31% of clones were deleted of the whole 5' part of their genome and were flanked, in 5', by the additional att sequence and, in 3', by an LTR. Integration of these last proviruses was often imprecise with respect to the viral ends. At total, 77% of proviruses had lost the packaging signal and were not mobilizable by a replication-competent virus and 92% had lost the selectable gene in a single round of replication. Although still to improve, the att vector could be considered as an interesting new safe retroviral vector for gene transfer experiments. PMID- 18420299 TI - Characterization of the genome sequence of an oncolytic Newcastle disease virus strain Italien. AB - We determined the complete genome sequence of strain Italien, a virulent and oncolytic strain of Newcastle disease virus (NDV) by direct nucleotide sequencing of RT-PCR products, a size of 15,186 nucleotides (nt). Comparison of six coding genes and non-coding regions of Italien with those of the other 25 sequenced strains revealed NDV Herts/33 was the most similar strain with Italien. The gene encoding the RNA dependent RNA polymerase was the most highly conserved, while the gene encoding phosphoprotein was the most highly variable. The HN and F proteins of Italien have been modeled on the crystal structure in order to study the structural characteristics. Interaction between the HN protein and the heptad repeat B (HRB) region of F protein was analyzed in silico by molecular docking predicted five critical residues I133, V142, D143, R480, and K567 on HN protein. Identification of amino acid residues that could be crucial for this interaction provides working hypotheses for subsequent studies. PMID- 18420301 TI - The accuracy of population health data for monitoring trends and outcomes among women with diabetes in pregnancy. AB - AIM: To assess the accuracy of routinely collected population birth and hospital datasets in identifying maternal pregestational diabetes mellitus (PDM) and gestational diabetes mellitus (GDM). METHODS: Information on maternal diabetes status was obtained from the medical records of a random sample of 1200 women and compared with routinely collected, population-based birth and hospital data. PDM and GDM are reported in both databases. Sensitivity, specificity, positive predictive value (PPV), negative predictive value and the kappa statistic were determined. RESULTS: Medical records were available for 1184 of the 1200 women sampled. 0.3% of women were classified with PDM and 4.8% with GDM. 'True' PDM was under-reported and misclassified in the birth data, but all cases were reported in the hospital data. GDM was also more completely and more accurately reported in the hospital data than in the birth data. Diabetes requiring insulin was more likely to be reported than non-insulin dependent diabetes. CONCLUSIONS: Hospital data were more sensitive and accurate (higher PPVs) than birth data and these measures were not improved by ascertaining diabetes from either of the two datasets. More severe forms of diabetes were more likely to be reported than less severe. PMID- 18420300 TI - Molecular epidemiology of rabies in Indonesia. AB - In order to clarify the genetic relationships and dynamics of rabies viruses that are epidemic in Indonesia, we determined and analyzed 1307 nucleotides of nucleoprotein genes of 34 rabies field isolates collected from Sumatra, Java, Kalimantan, Sulawesi and Flores islands. Results of phylogenetic analysis indicated that rabies isolates in Indonesia formed one cluster, were of Asian lineage, and were closely related to a rabies isolate in China rather than to rabies isolates in Thailand, India or Sri Lanka. Rabies isolates in Indonesia were divided into three phylogroups (ID1, ID2 and ID3) that included seven lineages. There was a correlation between phylogroup and geographical distribution of the isolates. Isolates in four lineages (SC1, SC2, SC3 and ST) of the ID1 phylogroup were mainly present in Sumatra. Isolates in the ST lineage were distributed widely in Sumatra, while isolates in the SC1, SC2 and SC3 lineages were limited to central Sumatra. ID2 and ID3 phylogroups included one lineage (JA) and two lineages (KS and SF), respectively. Results of phylogenetic analysis and historical background suggest that rabies viruses in China might have been transferred to Indonesia and spread to each island due to human activities. PMID- 18420302 TI - A heuristic index for selecting similar categories in multiple correspondence analysis applied to living donor kidney transplantation. AB - This work introduces a heuristic index (the "tolerance distance") to define the "closeness" of two variable categories in multiple correspondence analysis (MCA). This index is a weighted Euclidean distance where weightings are based on the "importance" of each MCA axis, and variable categories were considered to be associated when their distances were below the tolerance distance. This approach was applied to a renal transplantation data. The analysed variables were allograft survival and 13 of its putative predictors. A bootstrap-based stability analysis was employed for assessing result reliability. The method identified previously detected associations within the database, such as that between race of donors and recipients, and that between HLA match and Cyclosporine use. A hierarchical clustering algorithm was also applied to the same data, allowing for interpretations similar to those based on MCA. The defined tolerance distance could thus be used as an index of "closeness" in MCA, hence decreasing the subjectivity of interpreting MCA results. PMID- 18420303 TI - Laboratory investigation of flux reduction from dense non-aqueous phase liquid (DNAPL) partial source zone remediation by enhanced dissolution. AB - This study investigated the benefits of partial removal of dense nonaqueous phase liquid (DNAPL) source zones using enhanced dissolution in eight laboratory scale experiments. The benefits were assessed by characterizing the relationship between reductions in DNAPL mass and the corresponding reduction in contaminant mass flux. Four flushing agents were evaluated in eight controlled laboratory experiments to examine the effects of displacement fluid property contrasts and associated override and underride on contaminant flux reduction (R(j)) vs. mass reduction (R(m)) relationships (R(j)(R(m))): 1) 50% ethanol/50% water (less dense than water), 2) 40% ethyl-lactate/60% water (more dense than water), 3) 18% ethanol/26% ethyl-lactate/56% water (neutrally buoyant), and 4) 2% Tween-80 surfactant (also neutrally buoyant). For each DNAPL architecture evaluated, replicate experiments were conducted where source zone dissolution was conducted with a single flushing event to remove most of the DNAPL from the system, and with multiple shorter-duration floods to determine the path of the R(j)(R(m)) relationship. All of the single-flushing experiments exhibited similar R(j)(R(m)) relationships indicating that override and underride effects associated with cosolvents did not significantly affect the remediation performance of the agents. The R(j)(R(m)) relationship of the multiple injection experiments for the cosolvents with a density contrast with water tended to be less desirable in the sense that there was less R(j) for a given R(m). UTCHEM simulations supported the observations from the laboratory experiments and demonstrated the capability of this model to predict R(j)(R(m)) relationships for non-uniformly distributed NAPL sources. PMID- 18420304 TI - The area composita of adhering junctions connecting heart muscle cells of vertebrates. VI. Different precursor structures in non-mammalian species. AB - Recent studies on the formation and molecular organization of the mammalian heart have emphasized the architectural and functional importance of the adhering junctions (AJs), which are densely clustered in the bipolar end regions (intercalated disks, IDs) connecting the elongated cardiomyocytes of the adult heart. Moreover, we learned from genetic studies of mutated AJ proteins that desmosomal proteins, which for the most part are integral components of ID specific composite AJs (areae compositae, AC), are essential in heart development and function. Developmental studies have shown that the bipolar concentration of cardiomyocyte AJs in IDs is a rather late process and only completed postnatally. Here we report that in the adult hearts of diverse lower vertebrates (fishes, amphibia, birds) most AJs remain separate and distinct in molecular character, representing either fasciae adhaerentes, maculae adhaerentes (desmosomes) or- less frequently--some form of AC. In the mature hearts of the amphibian and fish species examined a large proportion of the AJs connecting cardiomyocytes is not clustered in the IDs but remains located on the lateral surfaces where they appear either as puncta adhaerentia or as desmosomes. In many places, these puncta connect parallel cardiomyocytes in spectacular ladder-like regular arrays (scalae adhaerentes) correlated with--and connected by--electron-dense plaque like material to sarcomeric Z-bands. In the avian hearts, on the other hand, most AJs are clustered in the IDs but only a small proportion of the desmosomes appears as AC, compared to the dominance of distinct fasciae adhaerentes. We conclude that the fusion and amalgamation of AJs and desmosomes to ACs is a late process both in ontogenesis and in evolution. The significance and possible functional implications of the specific junctional structures in vertebrate evolution and the class-specific requirements of architectural and molecular assembly adaptation during regeneration processes are discussed. PMID- 18420305 TI - Impact of Medicaid cutbacks on emergency department use: the Oregon experience. AB - STUDY OBJECTIVE: Federal policy changes and tightened state budgets may reduce Medicaid enrollment in many states. In March 2003, the Oregon Health Plan (Oregon's Medicaid expansion program) made substantial changes in its benefit package that resulted in the disenrollment of more than 50,000 beneficiaries. We sought to study the impact of these Oregon Health Plan policy changes on statewide emergency department (ED) use. METHODS: In this observational study, hospital billing data on 2,680,954 visits to 26 Oregon EDs were obtained, sampled up to 24 months before and 24 months after the cutbacks. These visits represent approximately 62% of all visits to Oregon's 58 EDs. We ascertained counts of ED visits by payer group before and after the Oregon Health Plan cutback date, plus hospital admissions from the ED as a measure of acuity. RESULTS: After the Oregon Health Plan policy changes, ED visits by the uninsured underwent an abrupt and sustained increase, from 6,682 per month in 2002 to 9,058 per month in 2004. Oregon Health Plan-sponsored and commercially insured visits decreased, resulting in a slight decrease in overall ED visits. Multivariable models adjusting for secular trends and seasonality showed a 20% (95% confidence interval 13% to 28%) increase in uninsured ED visits, whereas the adjusted number of Oregon Health Plan-sponsored visits decreased. The proportion of uninsured ED visits resulting in hospital admission increased (odds ratio 1.50; 95% confidence interval 1.39 to 1.62). CONCLUSION: Oregon's Medicaid cutbacks were followed by increases in ED use and hospitalizations by the uninsured. Recent federal legislation facilitating similar Medicaid changes in other states may lead to replication of these events elsewhere. PMID- 18420306 TI - Desensitization and internalization of the human motilin receptor is independent of the C-terminal tail. AB - The motilin receptor (MTLR) is an important therapeutic target for the treatment of hypomotility disorders but desensitization may limit its clinical utility. The aim of this study was to investigate the role of the C-terminal tail of the MTLR in the desensitization, phosphorylation and internalization process. Three MTLR mutants, C-terminally truncated from amino acid 412 till 384 (MTLRDelta385), 374 (MTLRDelta375) or 368 (MTLRDelta369), were constructed and C-terminally tagged with an EGFP and stably expressed in CHO cells co-expressing the Ca(2+) indicator apoaequorin. Activity and desensitization were studied by measuring changes in motilin-induced luminescent Ca(2+) rises. Receptor phosphorylation was investigated by immunoprecipitation and MTLR-EGFP internalization was visualized by fluorescence microscopy. Truncation only reduced MTLR affinity and the efficacy to induce Ca(2+) luminescent responses of the MTLRDelta375-EGFP mutant. Furthermore, the region between amino acid 375 and 368 seems to be important for proper cell surface expression of the MTLR since receptors of the MTLRDelta369 EGFP mutant but not of the other mutants were found intracellularly in vesicles. Truncation of the receptor till amino acid 384 or 374 did neither affect desensitization nor internalization. In contrast phosphorylation of the MTLRDelta385-EGFP mutant was reduced by 80% but was not affected in the MTLRDelta375-EGFP mutant. In conclusion, MTLR desensitization and internalization is not dependent on the presence of the C-terminal tail. Truncation favors internalization via either phosphorylation-independent pathways or via phosphorylation of alternative sites in the receptor. PMID- 18420307 TI - Hemodynamic-independent anti-natriuretic effect of urotensin II in spontaneously hypertensive rats. AB - The present study aims to test the hypothesis that U-II might have a direct anti natriuretic action in spontaneously hypertensive rats (SHR). Bolus U-II injection (15 nmol kg(-1)) caused a transient decrease in glomerular filtration rate (GFR), urine flow rate (UV), urinary sodium (UNaV) and potassium excretion (U(K)V) that corresponded with a committed decrease in mean arterial pressure (MAP) and renal blood flow (RBF) during the first 30 min. Continuous U-II infusion (0.2 nmol kg( 1)h(-1)) following a bolus U-II injection (0.3 nmol kg(-1)) caused an anti natriuretic effect without any significant change in MAP, RBF, GFR, UV and UKV during the entire 1.5-h perfusion period in SHR. The levels of aldosterone and angiotensin II were not altered in the plasma and kidney, while plasma antidiuretic hormone decreased in response to U-II injection (15 nmol kg(-1)). Protein levels of U-II receptors (UT) were significantly increased in the kidney of 17-week-old SHR when compared with the age-matched WKY rats, while mRNA transcripts of both U-II and UT were increased in the kidney, left ventricle and thoracic aorta. In conclusion, U-II exerts a hemodynamic-independent anti natriuretic action in adult SHR. The anti-natriuretic action of U-II in SHR is probably associated with an increased expression of the U-II-UT system in the kidney, suggesting a potential renal role of U-II in the pathogenesis of hypertension. PMID- 18420308 TI - PITX3 polymorphism is associated with early onset Parkinson's disease. AB - PITX3 is a transcription factor of importance for the differentiation and survival of midbrain dopaminergic neurons, the gene of which is disrupted in a putative mouse model for Parkinson's disease (PD). The A-allele of a HapMap tagging SNP (rs4919621) that was genotyped in a population of 361 PD patients, 69 of which had early onset, and in 333 controls, was significantly more common in PD patients with an early age of onset when compared either to controls (p=0.002) or to PD patients with late onset (p=0.001). In contrast, a previous finding suggesting a SNP (rs3758549) in the putative promoter region of the PITX3 gene to be associated with PD could not be replicated. PMID- 18420309 TI - Synthesis of new series of 5,6-dihydro-4H-1,2-oxazines via hetero Diels-Alder reaction and evaluation of antimicrobial activity. AB - A new series of 5,6-dihydro-4H-1,2-oxazines were synthesized via hetero Diels Alder reaction of alpha-nitrosoolefins with alkenes. alpha-Nitrosoolefins were generated from ketoximes by the action of chloramine-T. The newly synthesized compounds were characterized with IR, NMR, elemental analysis and screened for their antimicrobial activity; they exhibited excellent antimicrobial activity. The minimal inhibitory concentration of the compounds was in the range of 10-35 microg ml(-1) for bacteria and 10-40 microg ml(-1) for fungi. PMID- 18420310 TI - Synthesis and anti-HIV studies of 2- and 3-adamantyl-substituted thiazolidin-4 ones. AB - A series of novel thiazolidin-4-ones bearing a lipophilic adamantyl substituent at position 2 or 3 were synthesized. A majority of them showed a modest anti-HIV 1 activity, whereas 2-adamantan-1-yl-3-(4,6-dimethylpyrimidin-2-yl)-thiazolidin-4 one (8) was endowed with a remarkable antiviral potency (EC(50)=0.67 microM). The new series of compounds (22-29) with an adamantyl moiety at the 3-position of the thiazolidinone ring showed good to modest anti-HIV-1 activity (EC(50)=1.0-11 microM) but also pronounced cytostatic activity. For example 24, 26 and 29 showed an EC(50) of 1.0-2.0 microM, while the 50% effective concentrations for 23 and 28 were 7.8 and 11.0 microM, respectively. X-ray studies and quantum chemical calculations revealed that the anti-HIV activity of the compounds strongly depends on their dipole moments and conformation of the thiazolidinones. PMID- 18420311 TI - QSAR modelling of pancreatic beta-cell KATP channel openers R/S-3,4-dihydro-2,2 dimethyl-6-halo-4-(substituted phenylaminocarbonylamino)-2H-1-benzopyrans using MLR-FA techniques. AB - Potassium (K(+)) channel openers are a diverse group of compounds which are used for the treatment of diseases like angina pectoris, hypertension, congestive heart failure, anti-hypoglycemic (insulinoma), bronchial asthma, etc. R/S-3,4 dihydro-2,2-dimethyl-6-halo-4-(substituted phenylaminocarbonylamino)-2H-1 benzopyrans are a new series of ATP-sensitive potassium (K(ATP-pbeta)) channel openers selective towards pancreatic beta-cells. QSAR modelling was done on these series of compounds to find a more active and selective K(ATP-pbeta) channel opener selective towards beta-cells of pancreatic tissues. Wang-Ford charges, partition coefficient, molar refractivity, principle moment of inertia at X, Y and Z axes are used as predictor variables and logarithm of percentage of residual insulin secretion is treated as response variable for the modelling. Multiple linear regressions with factor analysis were performed to develop QSAR models. Four equations were obtained using different combinations of the predictor variables based on factor loadings. Regression coefficients of all descriptors used are significant at more than 95% level. Results showed that Wang Ford charges on atom numbers 11, 17, 18, 19 and 21 are important for the inhibition of residual insulin secretion. The presence of electron withdrawing group at m- and p-position of phenyl ring B is required for the inhibition. The energy minimized geometry of the most active compound supported our modelling. PMID- 18420312 TI - Branched and linear lipopeptide vaccines have different effects on primary CD4+ and CD8+ T-cell activation but induce similar tumor-protective memory CD8+ T-cell responses. AB - We compared murine T-cell responses to synthetic lipopeptide vaccines in which the TLR2 ligand Pam(2)Cys was attached to co-linear CD4+ and CD8+ T-cell epitopes of ovalbumin (OVA) in a linear or branched configuration. Mice received OVA specific transgenic CD8+ and CD4+ T-cells followed by one injection of vaccine. Although the branched lipopeptide was more potent in activating OVA-specific CD4+ and CD8+ T-cells in the primary response, both vaccines induced cytolytic T lymphocytes (CTL) that expressed perforin, granzyme A-C, and IFN-gamma mRNAs and conferred long-term protection of most mice against challenge with OVA-expressing tumor cells. OVA epitope display was reduced in tumors that developed in some mice, suggesting CD8+ T-cell dependent selection. PMID- 18420313 TI - What can HIV vaccine trials teach us about future HIV vaccine dissemination? AB - This investigation explored commonalities and differences in barriers and motivators to HIV vaccine trial participation and acceptability of future U.S. Food and Drug Administration (FDA)-approved HIV vaccines in order to identify implications of clinical trials for future HIV vaccine dissemination. Fifteen focus groups were conducted with 157 predominately ethnic minority and low income participants recruited using venue-based sampling in Los Angeles. Data were analyzed using narrative thematic analysis. Barriers and motivators in common across willingness to participate (WTP) in HIV vaccine trials and future HIV vaccine acceptability (e.g., concerns about vaccine-induced infection, false positives, side effects, efficacy, mistrust and stigma) suggest clinical trials present significant opportunities to develop and evaluate empirically based interventions to support future HIV vaccine dissemination. Barriers specific to HIV vaccine acceptability (e.g., concerns about duration of protection, cross clade protection, cost and access) also indicate the need for formative research focused specifically on future dissemination. Protection motivation, common to WTP and acceptability, highlights the need to provide and evaluate prevention counseling and education in clinical trials, which may form the basis of evidence informed preventive interventions to be launched in tandem with dissemination of partial efficacy HIV vaccines. PMID- 18420314 TI - Lung transplant patients' T cell responses to influenza vaccine viruses between seasons. AB - We hypothesized that T cell responses to influenza viruses by lung transplant and healthy individuals would be similar. Twelve lung transplant patients and 12 healthy individuals received influenza vaccines during two seasons. Lymphocytes were isolated for the trans-vivo delayed-type hypersensitivity assay from blood samples following immunization. T cell responses to all three inactivated influenza antigens from each were similar between the groups. However, the response to the A/New Caledonia (H1N1) virus by transplant patients was greater than the healthy controls' response, but only in the first season. The magnitude of the T cell responses in lung transplant patients is similar to those of healthy control individuals. These responses may be important in T cell memory. PMID- 18420315 TI - Impact of different case definitions for acute otitis media on the efficacy estimates of a pneumococcal conjugate vaccine. AB - Considerably higher vaccine efficacy estimate for clinical acute otitis media (AOM) has been obtained for the 11-valent pneumococcal conjugate vaccine with protein D of Haemophilus influenzae as a carrier (PncPD11) in the POET study than for the 7-valent pneumococcal conjugate vaccine (PncCRM7) in the Finnish Otitis Media (FinOM) Vaccine Trial. We recalculated PncCRM7 efficacy from the FinOM data using a case definition for AOM very close to the POET definition and a definition giving an incidence for AOM in the control group comparable to that obtained in the POET study. The different case definitions had only a slight impact on the vaccine efficacy estimates compared to the original case definitions. We were not able to show that the differences between the study results would be due to the case definitions used. PMID- 18420316 TI - Report of the third meeting on "influenza vaccines that induce broad spectrum and long-lasting immune responses", World Health Organization, Geneva, Switzerland, 3 4 December 2007. PMID- 18420317 TI - Performance of a commercial radon-in-water measurement kit. AB - Methods currently approved for the measurement of radon ((222)Rn) in water in New York State are liquid scintillation counting and emanation into alpha scintillation cells. A passive system using an electret ion chamber (EIC) was evaluated as an alternative for the measurement of radon in water. Over 130 water samples from a community water supply containing 32BqL(-1) and 30 standards containing 686BqL(-1) were measured using the EIC method over 1- to 4-day exposure times. For comparison, identical samples were measured using liquid scintillation counting. Results of duplicate samples were typically within 5% for liquid scintillation counting and within 10% for the EIC. With respect to accuracy, the EIC produced results that were consistently low by 11-15%. PMID- 18420318 TI - Interdisciplinary management of unfavorable posterior intermaxillary space. AB - The patient was a case of severe dentoalveolar extrusion of the right maxillary posterior segment due to early loss of mandibular molars. Therefore, it is difficult to replace the missing teeth with either fixed or removable prostheses. Her occlusion was Angle Class II malocclusion and no mandibular molars for anchorage on the right side. A posterior maxillary subapical osteotomy was utilized for reestablishing the intermaxillary space. A titanium alloy miniplate placed in the right mandible provide skeletal anchorage to distalize the anterior mandibular teeth. Dental implants were placed to restore masticatory function. PMID- 18420319 TI - Squamous cell carcinoma arising in the lining of an epidermoid cyst within the sublingual gland--a case report. AB - We present the case of squamous cell carcinoma that arose in the lining of an epidermoid cyst that developed within the sublingual gland in a 64-year-old woman. Malignant transformation in epidermoid cysts is rare, particularly in the floor of the mouth. We know of no cases reported of an epidermoid cyst arising within the sublingual gland. PMID- 18420320 TI - Random control trial of dermis-fat graft and interposition of temporalis fascia in the management of temporomandibular ankylosis in children. AB - Temporomandibular ankylosis is a disabling condition that affects hygiene and cosmetic appearance. Several interpositional grafts such as meniscus, muscle, fascia, skin, cartilage, fat, dura, alloplastic materials and xenografts have been used to prevent recurrence of ankylosis. We studied the advantages and disadvantages of dermis fat graft as an interposition material after arthroplasty and compared it with temporalis fascia interposition. Seventeen patients with temporomandibular ankylosis involving 20 joints were randomly divided into two groups; the first group had operations for interposition of dermis-fat graft that was taken from the groin. Patients in control group had operations to interpose temporalis fascia and muscle from the same surgical site. All were assessed by age, sex, etiology, clinical features and post surgical complications. The groups were matched in age and the male: female ratio was 0.89:1.The median duration of ankylosis was 7.3 (range 2-11) years. Postoperative and follow up interincisal mouth opening was satisfactory with good healing of the dermis-fat graft donor site. We conclude that the use of dermis fat grafts has minimal donor site morbidity, and is a safe and effective interposition material to prevent the recurrence of temporomandibular ankylosis. PMID- 18420321 TI - Panniculitis ossificans traumatica: an unusual presentation. AB - We describe a patient with panniculitis ossificans traumatica that arose adjacent to the mandible. Panniculitis ossificans traumatica is an unusual form of heterotopic calcification that develops within subcutaneous fat as a result of injury. There have been few reported cases, and we know of none in the maxillofacial region despite the frequency of trauma to this region. As a result few treatment protocols exist. We describe an experience with its diagnosis and management. PMID- 18420322 TI - A scientific agenda for the concept of recovery as it applies to schizophrenia. AB - Recovery is now a widely discussed concept in the field of research, treatment, and public policy regarding schizophrenia. As it has increasingly become a focus in mainstream psychiatry, however, it has also become clear both that the concept is often used in multiple ways, and that it lacks a strong scientific basis. In this review, we argue that such a scientific basis is necessary for the concept of recovery to have a significant long-term impact on the way that schizophrenia is understood and treated. The discussion focuses on key issues necessary to establish this scientific agenda, including: 1) differences in definitions of recovery and their implications for studying recovery processes and outcomes; 2) key research questions; 3) the implications of data from outcome studies for understanding what is possible for people diagnosed with schizophrenia; 4) factors that facilitate recovery processes and outcomes, and methods for studying these issues; and 5) recovery-oriented treatment, including issues raised by peer support. Additional conceptual issues that have not received sufficient attention in the literature are then noted, including the role of evidence-based practices in recovery-oriented care, recovery-oriented care for elderly people with schizophrenia, trauma treatment and trauma-informed care, and the role of hospitals in recovery-oriented treatment. Consideration of these issues may help to organize approaches to the study of recovery, and in doing so, improve the impact of recovery-based initiatives. PMID- 18420323 TI - Health beliefs and perceived need for mental health care of anxiety and depression--the patients' perspective explored. AB - Patients' illness representations and beliefs about treatment for depression and anxiety, as well as their perceived needs, are important for treatment. A systematic review was conducted of 71 studies describing the beliefs or perceived needs of patients and non-patients. Patients give multi-dimensional explanations for depression and see both psychological and medication treatment as helpful. People who suffer from depression have more positive beliefs about biological etiology and medication treatment than healthy people, or those with less severe depressive symptoms. Anxiety patients view psychological interventions as their best treatment option. Between 49% and 84% of the patients with depression or anxiety perceive a need for treatment, mostly for counseling and medication. All patients prefer psychological treatment forms to medication. A majority of patients view antidepressants as addictive and many perceive stigma and see practical and economic barriers to care. The most vulnerable groups in terms of seeking and receiving mental health care for depression and anxiety seem to be minority groups, as well as younger and older patients. More research is required into the specific needs of anxiety and depression patients. Open communication between patient and provider could lead to valuable improvements in treatment. PMID- 18420324 TI - Reducing child mortality in Nigeria: a case study of immunization and systemic factors. AB - The purpose of the study is to assess the outcome of the Expanded Program on Immunization (EPI) in Nigeria, as well as to examine systemic factors influencing its high under-five mortality rate (UFMR). The principal objective of the EPI program when it was implemented in 1978 was to reduce mortality, morbidity and disability associated with six vaccine preventable diseases namely tuberculosis, tetanus, diphtheria, measles, pertussis and poliomyelitis. The methodological approach to this study is quantitative, using secondary time series data from 1970 to 2003. The study tested three hypotheses using time series multiple regression analysis with autocorrelation adjustment as a statistical model. The results showed that the EPI program had little effect on UFMR in Nigeria. Only the literacy rate and domestic spending on healthcare had statistically significant effects on the UFMR. The military government was not a significant factor in reducing or increasing the UFMR. It appears that Nigeria needs a unified approach to healthcare delivery, rather than fragmented programs, to overcome cultural and political divisions in society. PMID- 18420325 TI - 'Face' and the embodiment of stigma in China: the cases of schizophrenia and AIDS. AB - The majority of theoretical models have defined stigma as occurring psychologically and limit its negative effects to individual processes. This paper, via an analysis of how 'face' is embodied in China, deepens an articulation of how the social aspects of stigma might incorporate the moral standing of both individual and collective actors defined within a local context. We illustrate (1) how one's moral standing is lodged within a local social world; (2) how one's status as a 'moral' community member is contingent upon upholding intrapersonal and social-transactional obligations; and (3) how loss of face and fears of moral contamination might lead to a 'social death'. We first draw from Chinese ethnographies that describe the process of human cultivation before one can achieve fully 'moral' status in society. We integrate findings from empirical studies describing how social-exchange networks in China are strictly organized based on the reciprocation of favors, moral positioning, and 'face'. We further ground these Chinese constructs within a theoretical framework of different forms of capital, and discuss the severe social consequences that loss of face entails. By utilizing the examples of schizophrenia and AIDS to illustrate how loss of moral standing and stigma is interwoven in China, we propose a model highlighting changes in moral status to describe how stigma operates. We suggest that symbolic restoration of moral status for stigmatized groups takes place as local-level stigma interventions. By analyzing the moral aspects of 'face', we propose that across cultures, stigma is embedded in the moral experience of participants, whereby stigma is conceived as a fundamentally moral issue: stigmatized conditions threaten what matters most for those in a local world. We further propose that stigma jeopardizes an actor's ability to mobilize social capital to attain essential social statuses. PMID- 18420326 TI - Corn oil enhancing hepatic lipid peroxidation induced by CCl4 does not aggravate liver fibrosis in rats. AB - Lipid peroxidation (LPO) is known to be associated with liver fibrosis in chronic liver injury. However, direct effects of the products of LPO on liver fibrogenesis have not been demonstrated. In this study, we examined the LPO products of carbon tetrachloride (CCl4)+corn oil to evaluate the effect of LPO products on liver fibrosis. CCl4 was given twice a week for 8 weeks. Corn oil was given daily to rats at a dose of 2 or 10ml/kg via gastrogavage throughout the whole experiment period. CCl4 induced both cyclooxygenase (COX)-2 independent and COX-2 dependent LPO. COX-2 independent LPO was enhanced by corn oil treatment while no effect was reflected on COX-2 dependent LPO. CCl4-induced liver fibrosis in rats was not aggravated by corn oil treatment. In addition, the amount of fatty liver induced by CCl4 was increased by corn oil treatment. Though the inflammation-related UCP-2 mRNA expression was induced by CCl4, it was not aggravated by the enhancement of corn oil. CONCLUSION: corn oil enriches polyunsaturated fatty acids through COX-2 independent pathways to increase LPO products that do not enhance liver fibrosis induced by CCl4. PMID- 18420327 TI - Horizontal gene transfer of virulence determinants in selected bacterial foodborne pathogens. AB - This review describes horizontal gene transfer from a historical point of view, with descriptions of the first instances of the different bacterial transfer mechanisms: conjugation, transduction and transformation, as well as examples of some of the early acknowledged transfer events. Gene transfer from four selected foodborne pathogens: Escherichia coli, Listeria monocytogenes,Staphylococcus aureus and Salmonella are highlighted. PMID- 18420328 TI - The inhalation toxicity of di- and triethanolamine upon repeated exposure. AB - Systemic and respiratory tract (RT) toxicity of triethanolamine (TEA) was assessed in a 28-day nose-only inhalation study in Wistar rats (10animals/sex, concentrations: 0, 20, 100, 500mg/m3; 5 days/week, 6h/day). In two nose-only 90 day inhalation studies, with similar exposure design, Wistar rats were exposed to 0, 15, 150, 400mg/m3 diethanolamine (DEA) (DEA Study 1:13animals/sex, general subchronic study) and to 0, 1.5, 3, 8mg/m3 (DEA Study 2:10animals/sex) to specifically investigate respiratory tract toxicity. Only DEA induced systemic toxicity at or above 150mg/m3 (body and organ weight changes, clinical- and histo pathological changes indicative for mild blood, liver, kidney and testicular effects). Neurotoxicity was not observed for both substances. Exposure to both substances resulted in laryngeal epithelial changes starting from 3mg/m3 for DEA (reversible metaplasia at the base of the epiglottis, inflammation at higher concentrations extending into the trachea) or from 20mg/m3 for TEA (focal inflammation, starting in single male animals). TEA appears to be less potent with respect to systemic toxicity and RT irritancy than DEA. The 90-day no adverse effect concentration" (NOAEC) for changes due to TEA exposure in the respiratory tract was 4.7mg/m3 derived by extrapolation from the NOAEC of the 28day study. PMID- 18420329 TI - The role of horizontal gene transfer in the evolution of selected foodborne bacterial pathogens. AB - Bacteria use various ways to transfer genetic information. These methods include: conjugation, which requires cell to cell contact between cells, transduction, which is bacteriophage-facilitated transfer of genetic information, and transformation, which is the uptake of free DNA from the environment. Usually the genes to be transferred lie on mobile genetic elements, pieces of DNA that encode proteins important to facilitate movement of DNA within or between genomes. This review highlights the transfer methods and the role of the assorted mobile genetic elements in the evolution of four foodborne bacterial pathogens: Escherichia coli O157:H7, Salmonella, Staphylococcus aureus and Listeria monocytogenes. PMID- 18420330 TI - Human exposure to PBDEs through the diet in Catalonia, Spain: temporal trend. A review of recent literature on dietary PBDE intake. AB - The mean concentrations of polybrominated diphenyl ethers (PBDEs) (sum tetra- to octaBDEs) were determined in samples of foodstuffs widely consumed by the population of Catalonia, Northeast Spain. The following six tetra-through heptabrominated congeners were also individually analyzed: PBDEs 47, 99, 100, 153, 154 and 183. Food samples were randomly acquired in 12 cities of Catalonia between March and June of 2006. The dietary intake of PBDEs was estimated for the population of this region. In order to determine the temporal trend on the exposure to PBDEs through the diet, the results were compared with those of a previous survey performed during 2000. The highest concentration of total PBDEs was found in fish and shellfish (563.9 ng/kg of wet weight), followed by oils and fats (359.3 ng/kg ww), and bakery products (98.8 ng/kg ww). Among six individually analyzed congeners, for most food groups BDE-47 and BDE-99 showed the highest levels. The dietary intake of PBDEs for a standard male adult of 70 kg body weight was 75.4 ng/day (or 1.1 ng/kg body weight/day, assuming ND=LOD/2). On a body weight basis, it means a decrease of 23% with respect to the daily intake of the 2000 survey (97.3 ng/day or 1.4 ng/kg/body weight/day). Finally, the current PBDE intake is compared with the results of recent studies on the dietary intake of PBDEs performed in various European, North American, and Asian countries. PMID- 18420331 TI - Effect of 2,4-dichlorophenoxyacetic acid on rat maternal behavior. AB - Exposure to 2,4-dichlorophenoxyacetic acid (2,4-D) has several deleterious effects on the nervous system such as alterations in the concentrations of neurotransmitters in the brain and/or behavioral changes, myelination rate, ganglioside pattern [Bortolozzi, A., Duffard, R., Antonelli, M., Evangelista de Duffard, A.M., 2002. Increased sensitivity in dopamine D(2)-like brain receptors from 2,4-dichlorophenoxyacetic acid (2,4-D)-exposed and amphetamine-challenged rats. Ann. N.Y. Acad. Sci. 965, 314-323; Duffard, R., Garcia, G., Rosso, S., Bortolozzi, A., Madariaga, M., DiPaolo, O., Evangelista de Duffard, A.M., 1996. Central nervous system myelin deficit in rats exposed to 2,4 dichlorophenoxyacetic acid throughout lactation. Neurotoxicol. Teratol. 18, 691 696; Evangelista de Duffard, A.M., Orta, C., Duffard, R., 1990. Behavioral changes in rats fed a diet containing 2,4-dichlorophenoxyacetic butyl ester. Neurotoxicology 11, 563-572; Evangelista de Duffard, A.M., Bortolozzi, A., Duffard, R.O., 1995. Altered behavioral responses in 2,4-dichlorophenoxyacetic acid treated and amphetamine challenged rats. Neurotoxicology 16, 479-488; Munro, I.C., Carlo, G.L., Orr, J.C., Sund, K., Wilson, R.M. Kennepohl, E. Lynch, B., Jablinske, M., Lee, N., 1992. A comprehensive, integrated review and evaluation of the scientific evidence relating to the safety of the herbicide 2,4-D. J. Am. Coll. Toxicol. 11, 559-664; Rosso et al., 2000], and its administration to pregnant and lactating rats adversely affects litter growth and milk quality. Since normal growth of the offspring depends on adequate maternal nursing and care, we evaluated the effect of 2,4-D on rat maternal behavior as well as the dam's monoamine levels in arcuate nucleus (AcN) and serum prolactin (PRL) levels. Wistar dams were exposed to the herbicide through the food from post partum day (PPD) 1 to PPD 7. Dams were fed either with a 2,4-D treated diet (15, 25 or 50mg 2,4-D/kg/daybw) or with a control diet. We observed that maternal nesting behavior was not modified by 2,4-D treatment. However, mother-pup interactions, specially the nursing behavior, were altered. Retrieval, crouching and licking of pups were reduced or suspended after 2,4-D treatment. We also observed an increase in the latency of retrieval and crouching in the dams treated with the herbicide. Dams showed movement along cage peripheries, food consumption during the light phase and high self-grooming. In addition of the deficits observed in maternal behavior parameters, increased catecholamine levels and a drastic decrease in indolamine levels in the AcN of treated dams were determined. Serum PRL levels were also diminished by 62%, 68% and 70% with respect to control dams in the 15, 25 and 50mg 2,4-D/kgbw treated dams, respectively. In conclusion, exposure to 2,4-D during the first post partum days produced changes in maternal behavior, serum prolactin and monoamine levels in the AcN of treated dams. PMID- 18420332 TI - A paradigm shift in endodontic management of immature teeth: conservation of stem cells for regeneration. AB - OBJECTIVE: This article will review the new concept of regenerative endodontics in the management of immature permanent teeth. The potential role of stem cells to regenerate immature permanent teeth after conservative treatment will be discussed. DATA AND SOURCES: Two sets of data source are focused in this review: (i) the characterization of various dental stem cells discovered since 2000 and (ii) recent clinical case reports showing that after conservative treatment, severely infected immature teeth with periradicular periodontitis and abscess can undergo healing and apexogenesis or maturogenesis. RESULTS: A new protocol of treating endodontically involved immature permanent teeth based on published articles to date is summarized in the review. The key procedures of the protocol are (1) minimal or no instrumentation of the canal while relying on a gentle but thorough irrigation of the canal system, (2) the disinfection is augmented with intra-canal medication of a triple-antibiotic paste between appointments, and (3) the treated tooth is sealed with mineral trioxide aggregate (MTA) and glass ionomer/resin cement at the completion of the treatment. Periodical follow-ups will take place to observe any continued maturation of the root. CONCLUSION: While more clinical research is needed, regenerative endodontics promotes a paradigm shift in treating endodontically involved immature permanent teeth from performing apexification procedures to conserving any dental stem cells that might remain in the disinfected viable tissues to allow tissue regeneration and repair to achieve apexogenesis/maturogenesis. PMID- 18420334 TI - Effects of dredging on benthic diatom assemblages in a lowland stream. AB - The objective of this study was to assess the effects of dredging on the structure and composition of diatom assemblages from a lowland stream and to investigate whether the response of diatom assemblages to the dredging is also influenced by different water quality. Three sampling sites were established in Rodriguez Stream (Argentina); physico-chemical variables and benthic diatom assemblages were sampled weekly in spring 2001. Species composition, cell density, diversity and evenness were estimated. Diatom tolerance to organic pollution and eutrophication were also analyzed. Differences in physico-chemical variables and changes in benthic diatom assemblages were compared between the pre and post-dredging periods using a t-test. Data were analyzed using Principal Components Analysis (PCA), non-metric multidimensional scaling (MDS) ordination and cluster analysis. The effects of dredging in the stream involve two types of disturbances: (i) in the stream bed, by the removal and destabilization of the substrate and (ii) in the water column, by generating chemical changes and an alteration of the light environment of the stream. Suspended solids, soluble reactive phosphorus and dissolved inorganic nitrogen were significantly higher in post-dredging periods. Physical and chemical modifications in the habitat of benthic diatoms produced changes in the assemblage; diversity and species numbers showed an immediate increase after dredging, decreasing at the end of the study period. Changes in the tolerance of the diatom assemblage to organic pollution and eutrophication were also observed as a consequence of dredging; in the post dredging period sensitive species were replaced by either tolerant or most tolerant species. These changes were particularly noticeable in site 1 (characterized by its lower amount of nutrients and organic matter previous to dredging), which showed an increase in the amount of nutrients and oxygen demand as a consequence of sediment removal. However, these changes were not so conspicuous in sites 2 and 3, which already presented a marked water quality deterioration before the execution of the dredging works. PMID- 18420333 TI - Antioxidant properties of resveratrol and piceid on lipid peroxidation in micelles and monolamellar liposomes. AB - The antioxidant activities of trans-resveratrol (trans-3,5,4'-trihydroxystilbene) and trans-piceid (trans-5,4'-dihydroxystilbene-3-O-beta-D-glucopyranoside), its more widespread glycosilate derivative, have been compared measuring their inhibitory action on peroxidation of linoleic acid (LA) and the radical scavenging ability towards different free radicals (such as DPPH) and radical initiators. It has been found that the two stilbenes have similar antioxidant capacity, while the comparison with BHT (2,6-di-tert-butyl-4-methylphenol) and alpha-tocopherol (vitamin E, vit. E), taken as reference, points out a slower but prolonged protective action against lipid peroxidation. Furthermore, piceid appears more efficacious than resveratrol as a consequence of the reaction of the latter with its radical form. The DSC profiles of phosphatidylcholine liposomes of various chain lengths, and EPR measurements of spin labelled liposomes demonstrated that the susceptible hydroxyl group of these compounds are located in the lipid region of the bilayer close to the double bonds of polyunsaturated fatty acids, making these stilbenes particularly suitable for the prevention and control of the lipid peroxidation of the membranes. PMID- 18420335 TI - Comparison of freehand 3-D ultrasound calibration techniques using a stylus. AB - In a freehand 3-D ultrasound system, a probe calibration is required to find the rigid body transformation from the corner of the B-scan to the electrical center of the position sensor. The most intuitive way to perform such a calibration is by locating fiducial points in the scan plane directly with a stylus. The main problem of this approach is the difficulty in aligning the tip of the stylus with the scan plane. The thick beamwidth makes the tip of the stylus visible in the B scan, even if the tip is not exactly at the elevational center of the scan plane. We present a novel stylus and phantom that simplify the alignment process for more accurate probe calibration. We also compare our calibration techniques with a range of styli. We show that our stylus and cone phantom are both simple in design and can achieve a point reconstruction accuracy of 2.2 mm and 1.8 mm, respectively, an improvement from 3.2 mm and 3.6 mm with the sharp and spherical stylus. The performance of our cone stylus and phantom lie between the state-of the-art Z-phantom and Cambridge phantom, where accuracies of 2.5 mm and 1.7 mm are achieved. PMID- 18420336 TI - Estimation of muscle fiber orientation in ultrasound images using revoting hough transform (RVHT). AB - Ultrasound imaging has been used frequently for the study of muscle contraction, including measurements of pennation angles and fascicle orientations. However, these measurements were traditionally conducted by manually drawing lines on the ultrasound images. In this study, we proposed a modified Hough transform (HT), aiming at automatically estimating orientations of straight line-shaped patterns, such as muscle fibers and muscle-bone interface in ultrasound images. The new method first located the global maximum in the HT accumulator matrix, which corresponded to the most dominant collinear feature points globally, using the standard HT; then the pixels close to the detected line were removed from the edge map, the HT accumulator matrix was calculated again, i.e., revoting, and a new line was detected. The iteration was repeated until the predefined termination conditions were satisfied. The performance of the algorithm was tested using computer-generated images with different levels of noises, as well as clinical ultrasound images, and compared with that of the conventional method. It was found that the orientation estimation results obtained by the new algorithm were well correlated (R2 = 0.965), with those obtained using the traditional method, i.e., drawing lines manually and reading the angles with the assistance of software. Further mean-difference plots revealed a difference of 0.18 +/- 2.41 degrees between the two methods at the 95% confidence level. Compared with the traditional method, the new algorithm was more capable of handling with highly noisy data and could avoid the aliasing problem, i.e., reporting multiple lines instead of single expected line. The results of this study suggested that the proposed revoting HT can be potentially used for the reliable and nonsubjective automatic estimation of the orientations of muscle fibers in musculoskeletal ultrasound images. PMID- 18420337 TI - Acoustic emissions during 3.1 MHz ultrasound bulk ablation in vitro. AB - Acoustic emissions associated with cavitation and other bubble activity have previously been observed during ultrasound (US) ablation experiments. Because detectable bubble activity may be related to temperature, tissue state and sonication characteristics, these acoustic emissions are potentially useful for monitoring and control of US ablation. To investigate these relationships, US ablation experiments were performed with simultaneous measurements of acoustic emissions, tissue echogenicity and tissue temperature on fresh bovine liver. Ex vivo tissue was exposed to 0.9-3.3-s bursts of unfocused, continuous-wave, 3.10 MHz US from a miniaturized 32-element array, which performed B-scan imaging with the same piezoelectric elements during brief quiescent periods. Exposures used pressure amplitudes of 0.8-1.4 MPa for exposure times of 6-20 min, sufficient to achieve significant thermal coagulation in all cases. Acoustic emissions received by a 1-MHz, unfocused passive cavitation detector, beamformed A-line signals acquired by the array, and tissue temperature detected by a needle thermocouple were sampled 0.3-1.1 times per second. Tissue echogenicity was quantified by the backscattered echo energy from a fixed region-of-interest within the treated zone. Acoustic emission levels were quantified from the spectra of signals measured by the passive cavitation detector, including subharmonic signal components at 1.55 MHz, broadband signal components within the band 0.3-1.1 MHz and low-frequency components within the band 10-30 kHz. Tissue ablation rates, defined as the thermally ablated volumes per unit time, were assessed by quantitative analysis of digitally imaged, macroscopic tissue sections. Correlation analysis was performed among the averaged and time-dependent acoustic emissions in each band considered, B-mode tissue echogenicity, tissue temperature and ablation rate. Ablation rate correlated significantly with broadband and low frequency emissions, but was uncorrelated with subharmonic emissions. Subharmonic emissions were found to depend strongly on temperature in a nonlinear manner, with significant emissions occurring within different temperature ranges for each sonication amplitude. These results suggest potential roles for passive detection of acoustic emissions in guidance and control of bulk US ablation treatments. PMID- 18420338 TI - Examining the time course of endothelium-independent dilation by nitroglycerin. AB - Arterial smooth muscle function is commonly assessed by measuring the dilation response to sublingual nitroglycerin (NTG), and is referred to as endothelium independent dilation (EID). The time point at which the brachial artery dilates maximally after NTG administration is generally believed to occur somewhere between three and five minutes. The aim of this study was to examine the time course of brachial artery dilation response to a single dose of sublingual NTG to identify the time point at which maximal dilation occurs and to assess factors effecting EID. Endothelial independent dilation was measured in 125 normal, healthy subjects (male = 67, female = 58) via high-resolution ultrasound on the brachial artery. Mean time to maximal EID was 279.0 +/- 26.9 s. There was a significant (p < 0.0001) difference between brachial EID at three minutes vs. five minutes post NTG. There was also a significant gender difference in EID response (p < 0.0001). The results of this study indicate that maximal EID occurs approximately five minutes after NTG administration. Measuring EID at three minutes after NTG administration likely underestimates the true dilation capacity of the artery. PMID- 18420339 TI - Laparoscopic augmentation ileocystoplasty: results and outcome. AB - BACKGROUND: Routine use of laparoscopic augmentation ileocystoplasty has not yet been established. OBJECTIVES: To assess the outcome of laparoscopic augmentation ileocystoplasty. DESIGN, SETTING, AND PARTICIPANTS: Twenty-three patients underwent laparoscopic augmentation ileocystoplasty for hypocompliant bladder. INTERVENTION: Bladder dissection and reconstruction of the ileovesical anastomosis were performed laparoscopically, whereas the ileal pouch was prepared extracorporeally through a small 3- to 4-cm muscle-splitting incision. MEASUREMENTS: Patient data, operative details, and follow-up were recorded. Urodynamic evaluation was performed preoperatively and after 12 mo, taking the bladder capacity and the maximum detrusor pressure as a measure for the outcome of the procedure. RESULTS AND LIMITATIONS: All cases were completed laparoscopically, with a mean operative time 202 min; mean hospital stay 5 d, and mean urethral catheter duration 11 d. After 12 mo, the estimated bladder volume increased from a mean 111 ml to 788 ml (p<0.01), whereas the maximum detrusor pressure dropped from a mean 92 cm H(2)O to 15 cm H(2)O (p<0.01). During a mean follow-up of 39 mo, two long-term complications have been reported: bladder stone and spontaneous rupture of the augmented bladder due to neglected clean intermittent self-catheterization. CONCLUSIONS: Laparoscopic augmentation ileocystoplasty is a safe procedure, technically feasible and with favourable urodynamic outcome. PMID- 18420340 TI - Editorial comment on: Laparoscopic augmentation ileocystoplasty: results and outcome. PMID- 18420341 TI - Origin of pluripotent germ cell tumours: the role of microenvironment during embryonic development. AB - Carcinoma in situ (CIS) testis, known also as intratubular germ cell neoplasia, is the cancer stem cell from which the great majority of testicular germ cell derived tumours (TGCTs) of the testis arise. TGCTs can proliferate into morphologically homogeneous seminomas or can differentiate into virtually any type of tissue and form teratomas (non-seminomas). CIS cells display a close phenotypic similarity to fetal germ cells (primordial germ cells or gonocytes) suggesting an origin due to a developmental delay or arrest of differentiation of early germ cells. The pluripotency of these neoplasms has recently been explained by a close resemblance of their expression profile to that of embryonic inner cell mass cells studied in culture as embryonic stem cells, with high expression of transcription factors associated with pluripotency, such as NANOG and OCT3/4, as well as proteins found in several tissue specific stem cells, such as TFAP2C (AP-2gamma) or KIT. CIS and seminomas highly express a number of pre-meiotic germ cell specific genes, which are down-regulated during development to non seminomas, while the expression of other embryonic markers, such as SOX2, is up regulated. The mechanistic pathways and causative factors remain to be elucidated of both the initial transformation of fetal germ cells into CIS cells and the progression of CIS cells into an invasive tumour in the young adult. However, evidence supported by epidemiological studies indicate that disturbances in the hormonal microenvironment of the differentiating gonads may results in both the neoplasia and a host of other problems later in life, such as genital malformations, decreased spermatogenesis, and signs of hypogonadism. PMID- 18420342 TI - Anti-angiogenic effect of 5-Fluorouracil-based drugs against human colon cancer xenografts. AB - In addition to the direct cytotoxic effects of chemotherapy agents on tumor cells, the anti-angiogenic activities attained by these agents by targeting proliferating endothelial cells in tumor blood vessels has attracted much research interest. In this study, we examined the antitumor activity of 5 Fluorouracil (5-FU)-based drugs (S-1 [1M tegafur, 0.4M 5-chloro-2,4 dihydroxypyridine and 1M potassium oxonate] and capecitabine) on human colorectal cancer xenografts and evaluated their anti-angiogenic effects. Both drugs showed significant antitumor activities against COL-1 xenografts at a sub-maximum tolerated dose (sub-MTD), which was lower than the maximum tolerated dose (MTD). At the sub-MTD, a significant reduction in the microvessel number and the enhancement of tumor-associated microvessel endothelial cell apoptosis was seen in xenografts treated with S-1. In addition, we found that thrombospondin-1 (TSP 1) expression, known to be a mediator of the anti-angiogenic effects of metronomic chemotherapy, was significantly up-regulated in xenograft tumor tissues and plasma in animals treated with S-1 at a sub-MTD. Capecitabine also showed a trend toward the induction of TSP-1. These results suggest that 5-FU based drugs inhibit tumor progression through different modes of action, including cytotoxic activity derived from 5-FU and the inhibition of angiogenesis through the induction of TSP-1. PMID- 18420343 TI - Solid-phase extraction method for preconcentration of trace amounts of some metal ions in environmental samples using silica gel modified by 2,4,6-trimorpholino 1,3,5-triazin. AB - A method was proposed for the preconcentration of some transition elements at trace levels using a column packed with silica gel modified by a synthetic ligand. Metal ions were adsorbed on 2,4,6-trimorpholino-1,3,5-triazin modified silica gel, then analytes retained on the adsorbent were eluted by 1molL(-1) hydrochloric acid and determined by flame atomic absorption spectrometry (FAAS). The influences of some experimental parameters including pH of the sample solution, weight of adsorbent, type, concentration and volume of eluent, flow rates of the sample solution and eluent, and sample volume on the preconcentration efficiency have been investigated. The influences of some matrix elements were also examined. The method also was used for simultaneous preconcentration of these elements and the method was successfully applied to the preconcentration and determination of them. The detection limits of the method for Ni2+, Co2+, Cd2+ and Zn2+were 0.29, 0.20, 0.23 and, 0.30ngmL(-1), respectively. The application of this modified silica gel to preconcentration of investigated cation from tap water, lake water, urine and apple leaves gave high accuracy and precision (relative standard deviation (R.S.D.) <3%). PMID- 18420344 TI - Degradation of quinoline by Rhodococcus sp. QL2 isolated from activated sludge. AB - A novel aerobic gram-positive bacterial strain capable of utilizing quinoline as sole source of carbon, nitrogen and energy was isolated from activated sludge of a coke plant wastewater treatment process. The isolate was identified as Rhodococcus sp. QL2 based on its morphology, physiochemical properties in addition to the results from 16S rDNA sequence analysis. The optimum temperature and the pH for its growth were 35-40 degrees C and 8.0, respectively. Extra nitrogen sources stimulated the bacterial growth on quinoline. Strain QL2 had strong quinoline degradability, and its degradation kinetics could be described with Haldane's model. Strain QL2 also had a broad range of substrate utilization. Identification of intermediates by GC/MS showed Rhodococcus sp. QL2 degraded quinoline via two pathways simultaneously. PMID- 18420345 TI - Decreased serum brain-derived neurotrophic factor in trained men. AB - The purpose of this study was to clarify the effect of physical activity on the level of serum brain-derived neurotrophic factor (BDNF). The serum BDNF level in trained men who have participated in regular sport activity (n=12) was compared to that in sedentary subjects (n=14). The physical activity levels expressed as total energy expenditure, move-related energy expenditure and walking count in the trained were significantly higher than those in the sedentary. The serum BDNF level in the trained men was found to be lower than that in the sedentary (19.54+/-4.53 ng/ml vs. 23.63+/-2.94 ng/ml, respectively, P<0.01). The serum BDNF level showed a significant negative correlation with daily total energy expenditure (r=-0.507, P<0.05), movement-related energy expenditure (r=-0.503, P<0.05), and walking count (r=-0.480, P<0.05). These results may suggest that vigorous habitual physical activity decrease the serum BDNF level. PMID- 18420346 TI - Effect of cytokines on neuronal excitability. AB - Numerous studies have shown that proinflammatory cytokines induce or facilitate pain and hyperalgesia in the presence of inflammation, injury to the nervous system or cancer. Besides acting as inflammatory mediators, increasing evidence indicates that cytokines may also specifically interact with receptor and ion channels regulating neuronal excitability, synaptic plasticity and injury under both physiological and pathological conditions. Here we summarize findings on two prototypical proinflammatory cytokines, tumor-necrosis factor-alpha and interleukin-1 beta, and their effects on neuronal excitability and ion channels with special regards to pain and hyperalgesia. PMID- 18420347 TI - Investigation of variants in the promoter region of PIK3C3 in schizophrenia. AB - The PIK3C3 gene has been implicated as a candidate gene for schizophrenia by functional evidence and genetic association studies. A series of previous studies have found susceptibility SNPs in promoter region. To further verify its susceptibility to schizophrenia in the Chinese population and the function of the polymorphisms, we performed a case control study in 556 unrelated schizophrenia patients and 563 normal controls as well as an in vitro functional analysis. In our association analysis of-432C-/T, we discovered obvious differences in allele frequency between patients and controls (P=0.017). A T/C haplotype constructed by -432C-/T and -86insC, which are tightly linked with each other (r(2)=1) can significantly weaken promoter's transcriptional activity by 20% (p=0.002 by t test). Though we cannot exclude the possibility that susceptibility of -432C-/T is caused by its linkage disequilibrium with other causal variants, our results do support PIK3C3 play a significant role in the etiology of schizophrenia. PMID- 18420348 TI - Weakened feedback abolishes neural oblique effect evoked by pseudo-natural visual stimuli in area 17 of the cat. AB - The psychological oblique effect, a well-known phenomenon that humans and some mammals are more visually sensitive to cardinal (vertical and horizontal) contours than to oblique ones, has commonly been associated with the overrepresentation of cardinal orientations in the visual cortex. In contrast to the oblique effect, however, Essock et al. [E.A. Essock, J.K. DeFord, B.C. Hansen, M.J. Sinai, Oblique stimuli are seen best (not worst!) broad-band stimuli: a horizontal effect, Vision Res. 43 (2003) 1329-1335] reported a psychological 'horizontal effect', in which visual stimuli dominated by oblique orientations were best perceived by human subjects when tested with unique natural broad-band stimuli. In this study, using optical imaging and the similar visual stimuli, we found an overrepresentation of cardinal orientations, i.e. the neural oblique effect, but not 'horizontal effect', in area 17 of the cat. In addition, the oblique effect was abolished by GABA administration in area 21a due to the preferred orientation shifting (6.0%) and decrease of orientation selectivity strength of neurons (26.9%) in area 17. These results indicate a neuronal basis of the oblique effect when animals watch a more natural scene, whereas no evidence was found for the 'horizontal effect'. PMID- 18420349 TI - Linearity of eprinomectin pharmacokinetics in lactating dairy sheep following pour-on administration: excretion in milk and exposure of suckling lambs. AB - Pharmacokinetics of eprinomectin (EPR) were studied in blood plasma and milk in two groups of six Istrian Pramenka dairy sheep and their suckling lambs following pour-on administration of EPR to ewes at dose levels of 0.5 and 1mg/kg. Maximum concentration in plasma was 2.22 and 5.25 microg/l, and AUC was 13.6 and 33.7 microg day/l for the 0.5 and 1.0mg/kg dose, respectively. These results indicate that drug exposure with a dose of 0.5mg/kg, which is commonly used in cattle, may be subtherapeutic. The concentration time course in milk paralleled plasma concentrations. In the dose range studied, linear pharmacokinetics of EPR were demonstrated. Milk-to-plasma AUC ratio was 0.79+/-0.12 and 1.12+/-0.43; the fraction of dose recovered in milk was 0.037+/-0.011 and 0.058+/-0.027% for the low and high dose, respectively. Maximum residual levels in milk were below the maximum acceptable level of 20 microg/kg; however, EPR was detected in all samples investigated. Despite low permeability in milk, AUC in plasma of suckling lambs was between 20 and 30% of the AUC in plasma of ewes. PMID- 18420350 TI - Strain differences in the effects of adrenalectomy on the midbrain dopamine system: implication for behavioral sensitization to cocaine. AB - Adrenalectomy (ADX) abolishes behavioral sensitization to cocaine in DBA/2, but not C57BL/6 inbred mice. The present study tests the hypothesis that this ADX effect on behavioral sensitization in the DBA/2 strain involves changes in midbrain dopamine systems that do not occur in the C57BL/6 strain. For that purpose, we have measured tyrosine hydroxylase (TH) and dopamine transporter (DAT) mRNA and D1- and D2-like receptor binding in C57BL/6 and DBA/2 mice that were i) unoperated, ii) ADX or sham (SHAM) operated, or iii) ADX or SHAM operated and subjected to a cocaine sensitization regimen (15.0 mg/kg cocaine on nine consecutive days, followed by a 7.5 mg/kg challenge after a 5-day withdrawal). ADX prevented behavioral sensitization to cocaine in the DBA/2, but not the C57BL/6 strain [de Jong IEM, Oitzl MS, de Kloet ER (2007) Adrenalectomy prevents behavioural sensitisation of mice to cocaine in a genotype-dependent manner. Behav Brain Res 177:329-339]. Mice were killed under basal conditions, in the latter case 24 h after the cocaine challenge. ADX did not affect the dopaminergic markers in drug naive mice. By contrast, strain-dependent neuroadaptations were found in the midbrain dopamine system of mice subjected to the sensitization regimen. In the DBA/2 strain, sensitization-resistant ADX mice were characterized by reduced D2 binding in the nucleus accumbens core and rostral caudate putamen. Furthermore, ADX prevented the increase in TH and DAT mRNA expression in the substantia nigra, and the decrease in D2 binding in the dorsomedial subdivision of the caudal caudate putamen associated with sensitization in SHAM mice. In the C57BL/6 strain ADX only marginally affected dopaminergic adaptations. These data suggest that adrenal hormones modulate behavioral sensitization to cocaine in a genotype-dependent fashion possibly through adaptations in pre- and post-synaptic components of the midbrain dopamine system. During cocaine sensitization, the DBA/2, but not the C57BL/6 strain, was susceptible to ADX in the dopamine system with respect to presynaptic TH and DAT and terminal D2 receptor expression. PMID- 18420351 TI - Alteration of dopaminergic markers in gastrointestinal tract of different rodent models of Parkinson's disease. AB - Parkinson's disease (PD) is a progressive neurological disorder that is often associated with various gastrointestinal (GI) symptoms. The link between the alteration of dopaminergic system and the symptoms of the GI tract in PD is complicated. To determine the changes in the dopaminergic system in the GI tract in PD, two kinds of rodent PD models were used in the present study. One was 6 hydroxydopamine (6-OHDA) -treated rats in which 6-OHDA was microinjected in the bilateral substantia nigra (SN). The other was 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine (MPTP) -treated mice in which MPTP was injected intraperitoneally. Immunofluorescence, reverse transcription (RT)-real time polymerase chain reaction (PCR) and Western blot were used to evaluate and compare the levels of mRNA and protein expression of tyrosine hydroxylase (TH) and dopamine transporter (DAT) in the GI tract between normal and rodent PD models, as well as between 6-OHDA-treated rats and MPTP-treated mice. The results indicated that TH- and DAT-positive cells were widely distributed in the GI tract. There were significant differences in TH and DAT expression in the GI tract between normal and PD models, as well as between 6-OHDA-treated rats and MPTP-treated mice. The protein levels of TH and DAT in the GI tract were significantly increased in 6-OHDA-treated rats, but the protein level of TH was significantly decreased in MPTP-treated mice. In addition, there was visible atrophy of gastric epithelial parietal cells in MPTP-treated mice, although the protein level of DAT was not significantly changed. The different alterations of dopaminergic system in the GI tract of the two kinds of PD models might underline the differences in GI symptoms in PD patients and might be correlated with the disease severity and disease process affecting the GI tract. PMID- 18420352 TI - Enhanced vascular permeability in rat skin induced by sensory nerve stimulation: evaluation of the time course and appropriate stimulation parameters. AB - Activation of nociceptors causes them to secrete neuropeptides. The binding of these peptides to receptors on blood vessels causes vasodilation and increased vascular permeability that allows loss of proteins and fluid (plasma extravasation, PE); this contributes to inflammation. This study defines the relationship between electrical activation of nociceptors and PE and evaluates the time course of this response in the skin of rats. We measured the time course and extent of PE by digital imaging of changes in skin reflectance caused by leakage of Evans Blue (EB) dye infused in the circulatory system before stimulation. Stimulation of the exclusively sensory saphenous nerve caused the skin to become dark blue within 2 min due to accumulation of EB. While PE is usually measured after 5-15 min of electrical stimulation, we found that stimulation for only 1 min at 4 Hz produced maximum PE. This response was dependent on the number of electrical stimuli at least for 4 Hz and 8 Hz stimulation rates. Since accumulation of EB in the skin is only slowly reversible, to determine the duration of enhanced vascular permeability we administered EB at various times after electrical stimulation of the saphenous nerve. PE was only observed when EB was infused within 5 min of electrical stimulation but could still be observed 50 min after capsaicin (1%, 25 microl) injection into the hind paw. These findings indicate that enhanced vascular permeability evoked by electrical stimulation persists only briefly after release of neuropeptides from nociceptors in the skin. Therefore, treatment of inflammation by blockade of neuropeptide release and receptors may be more effective than treatments aimed at epithelial gaps. We propose, in models of stimulation-induced inflammation, the use of a short stimulus train. PMID- 18420353 TI - First a hero of science and now a martyr to science: the James Watson Affair - political correctness crushes free scientific communication. AB - In 2007 James D. Watson, perhaps the most famous living scientist, was forced to retire from his position and retreat from public life in the face of international mass media condemnation following remarks concerning genetically caused racial differences in intelligence. Watson was punished for stating forthright views on topics that elite opinion has determined should be discussed only with elaborate caution, frequent disclaimers, and solemn deference to the currently-prevailing pieties. James Watson has always struck many people as brash; however this blunt, truth-telling quality was intrinsic to his role in one of the greatest scientific discoveries. Much more importantly than 'good manners', Watson has consistently exemplified the cardinal scientific virtue: he speaks what he understands to be the truth without regard for the opinion of others. The most chilling aspect of the Watson Affair was the way in which so many influential members of the scientific research community joined the media condemnation directed against Watson. Perhaps the most egregious betrayal of science was an article by editorialists of the premier UK scientific journal Nature. Instead of defending the freedom of discourse in pursuit of scientific truth, Nature instead blamed Watson for being 'crass' and lacking 'sensitivity' in discussing human genetic differences. But if asked to choose between the 'sensitive' editors of Nature or the 'crass' genius of James D. Watson, all serious scientists must take the side of Watson. Because when a premier researcher such as Watson is hounded from office by a vicious, arbitrary and untruthful mob; all lesser scientists are made vulnerable to analogous treatment at the whim of the media. A zealous and coercive brand of 'political correctness' is now making the biological truth of human genetic differences intolerably difficult to discover and discuss in US and UK. This needs to change. My hope is that truth will prevail over political correctness and James Watson will not just be exonerated but vindicated as an exemplar of the true morality of science: that scientific communication needs to be allowed to be clear, direct - even crass - in the pursuit of truth. James Watson has been a hero of science for the achievements of his career, and also a martyr for science at the end of his career. PMID- 18420354 TI - Cancer: a profit-driven biosystem? AB - The argument is made that solid malignant tumors behave as profit-driven biological systems in that they expand their nutrient-uptaking surface to increase energetic revenue, at a comparably low metabolic cost. Within this conceptual framework, cancer cell migration is a critical mechanism as it maximizes systemic surface expansion while minimizing diffusion distance. Treating these tumor systems with adjuvant anti-proliferative regimen only should increase the energetic net gain of the viable cancer cells left behind, hence would facilitate tumor recurrence. Therapeutic attempts to better control tumor (re)growth should therefore aim primarily at containing its surface expansion, thus reducing its energetic revenue, or increasing its metabolic costs or better yet, both. PMID- 18420355 TI - Alcohol drinking and renal cell carcinoma in Canadian men and women. AB - BACKGROUND: Epidemiologic studies have reported that moderate alcohol consumption is inversely associated with the risk of renal cell carcinoma (RCC), but sex specific results are inconsistent. The present study examines the association between alcohol intake and the risk of RCC among men and women. METHODS: Mailed questionnaires were completed by 1138 newly diagnosed, histologically confirmed RCC cases and 5039 population controls between 1994 and 1997 in eight Canadian provinces. A food frequency questionnaire provided data on eating habits and alcohol consumption 2 years before data collection. Other information included socio-economic status, lifestyle habits, alcohol use, and diet. Odds ratios (ORs) and 95% confidence intervals (CIs) were derived through unconditional logistic regression. RESULTS: Total alcohol intake was inversely associated with RCC in men and in women; the OR for the highest intake group (> or =22.3 g/day among men and > or =7.9 g/day among women) versus the non-drinkers was 0.7 (95% CI, 0.5 0.9) for both sexes. Analysis of menopausal status produced ORs for the highest intake group versus the non-drinkers of 1.2 (95% CI, 0.7-2.1) among premenopausal women and 0.6 (95% CI, 0.4-0.9) among postmenopausal women. Smoking and obesity were not important effect modifiers. CONCLUSION: Moderate alcohol consumption may be associated with a decreased risk of RCC in men and in women (mainly postmenopausal women). PMID- 18420356 TI - Protective properties of Leucas lavendulaefolia extracts against D-galactosamine induced hepatotoxicity in rat. AB - Different extracts of the leaves of Leucas lavendulaefolia were tested against D galactosamine (D-GalN) induced liver toxicity in rats. The methanol extract (100 mg/kg, p.o.) which exhibited significant hepatoprotective activity seems to support the claim of folk medicine. PMID- 18420358 TI - Eukaryotic origin of glyceraldehyde-3-phosphate dehydrogenase genes in Clostridium thermocellum and Clostridium cellulolyticum genomes and putative fates of the exogenous gene in the subsequent genome evolution. AB - Although lateral gene transfer (LGT) events have been frequently documented in the evolution of glyceraldehyde-3-phosphate dehydrogenase (GAPDH), no eukaryote to-prokaryote transfer has been reported so far. Here we describe the first case of the GAPDH gene transfer from a eukaryote to a subset of Clostridium species (Bacteria, Firmicutes). A series of phylogenetic analyses of GAPDH homologues revealed that Clostridium thermocellum and Clostridium cellulolyticum homologues have the evolutionary affinity to the eukaryotic homologues, rather than to those of bacterial species closely related to the two Clostridium species in the organismal phylogeny. These results suggest that the GAPDH genes in the two Clostridium species are of eukaryotic origin, which is the first reported case of eukaryote-to-bacterium GAPDH gene transfer. Since a previously published 16S ribosomal DNA phylogeny and our GAPDH phylogeny commonly suggest an intimate evolutionary relationship between C. thermocellum and C. cellulolyticum, a common ancestor of the two species likely acquired the eukaryotic GAPDH gene. In the C. cellulolyticum genome, the exogenous GAPDH gene was physically separated from other glycolytic genes, suggesting that this gene organization was likely achieved by a random insertion of the laterally transferred gene. On the other hand, in the C. thermocellum genome, the laterally transferred GAPDH gene clusters with other bacterial glycolytic genes. We discuss possible scenarios for the evolutionarily chimeric glycolytic gene cluster in the C. thermocellum genome. PMID- 18420359 TI - Questionable 16S ribosomal RNA gene annotations are frequent in completed microbial genomes. AB - According to recent reports, many ribosomal RNA gene annotations are still questionable, and the use of inappropriate tools for annotation has been blamed. However, we believe that the abundant 16S rRNA partial sequence in the databases, mainly created by culture-independent PCR methods, is another main cause of the ambiguous annotations of 16S rRNA. To examine the current status of 16S rRNA gene annotations in complete microbial genomes, we used as a criterion the conserved anti-SD sequence, located at the 3' end of the 16S rRNA gene, which is commonly overlooked by culture-independent PCR methods. In our large survey, 859 16S rRNA gene sequences from 252 different species of the microbial complete genomes were inspected. 67 species (234 genes) were detected with ambiguous annotations. The common anti-SD sequence and other conserved 16S rRNA sequence features could be detected in the downstream-intergenic regions for almost every questionable sequence, indicating that many of the 16S rRNA genes were annotated incorrectly. Furthermore, we found that more than 91.5% of the 93,716 sequences of the available 16S rRNA in the main databases are partial sequences. We also performed BLAST analysis for every questionable rRNA sequence, and most of the best hits in the analysis were rRNA partial sequences. This result indicates that partial sequences are prevalent in the databases, and that these sequences have significantly affected the accuracy of microbial genomic annotation. We suggest that the annotation of 16S rRNA genes in newly complete microbial genomes must be done in more detail, and that revision of questionable rRNA annotations should commence as soon as possible. PMID- 18420357 TI - DSM-IV criteria-based clinical subtypes of cannabis use disorders: results from the National Epidemiological Survey on Alcohol and Related Conditions (NESARC). AB - Prior research documented high homogeneity of alcohol use disorders (AUDs) as clinical entities. However, it is unknown whether this finding extends to other substance use disorders. We investigated this by examining the prevalence of all possible DSM-IV criteria-based clinical subtypes of current and lifetime cannabis use disorders in the general population. The number of possible (i.e., theoretical) clinical subtypes of cannabis abuse and dependence based on different combinations of the DSM-IV criteria was calculated using the combinatorial function. This number was compared with the subtypes actually observed in the National Epidemiologic Survey on Alcohol and Related Conditions (NESARC), a large U.S. national sample (N=43,093). Clinical and demographic correlates of the subtypes were examined with chi2 tests whose target population was the United States civilian non-institutionalized population. All DSM-IV cannabis abuse and dependence criteria were assessed with the Alcohol Use Disorder and Associated Disabilities Interview Schedule-DSM-IV Version (AUDADIS IV). Of all possible cannabis dependence subtypes, 29 (69%) were observed in the 12-month timeframe, and 41 (98%) in the lifetime timeframe. The corresponding numbers of subtypes for cannabis abuse were 12 (75%), current and 15 (100%), lifetime. These findings suggest that, in contrast to alcohol disorders, cannabis use disorders were highly heterogeneous. Future research should investigate whether there are differences in the course and treatment response of these clinical subtypes of cannabis use disorders, and the heterogeneity of other substance use disorders. PMID- 18420361 TI - Polymorphisms and haplotypes of DNA repair and xenobiotic metabolism genes and risk of DNA damage in Chinese vinyl chloride monomer (VCM)-exposed workers. AB - In this case-control study, we investigated the association between DNA damage and genetic susceptibility among vinyl chloride monomer (VCM)-exposed workers. The cumulative exposure dose of VCM was calculated based on the workers' duration of exposure and the geometric mean concentration of VCM in the workplace. DNA damage to peripheral blood lymphocytes was measured by single cell gel electrophoresis (SCGE) assay, and single nucleotide-polymorphisms (SNPs) in xenobiotic metabolism and DNA repair genes were detected by the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) methods. Univariate analysis showed that the CYP2E1 c1c2/c2c2 and XPD751 Lys/Gln and Gln/Gln genotypes were significantly associated with the levels of DNA damage (P<0.01 and 0.05, respectively). Further logistic regression analysis showed a significant association between CYP2E1 c1c2/c2c2 and DNA damage, and risk of having increased levels of DNA damage was more pronounced in those individuals having XRCC1 194 mutant genotypes and/or XPD751 Lys/Gln and Gln/Gln genotypes. Although most of the XPD and XRCC1 haplotypes did not show any significant difference, the XRCC1 haplotype Trp194-Arg280 was significantly over-represented in the case group (P<0.05; OR 2.09; 95% CI: 1.07-4.06) than in controls. Overall, our data suggest that the genotypes of CYP2E1, XRCC1 194, and XPD 751 were associated with the level of DNA damage and may contribute to individual sensitivity to DNA damage induced by VCM in the workplace. PMID- 18420360 TI - Inhaled ultrafine particulate matter affects CNS inflammatory processes and may act via MAP kinase signaling pathways. AB - In addition to evidence that inhalation of ambient particulate matter (PM) can increase cardiopulmonary morbidity and mortality, the brain may also constitute a site adversely effected by the environmental presence of airborne particulate matter. We have examined the association between exposure to PM and adverse CNS effects in apolipoprotein E knockout (ApoE-/-) mice exposed to two levels of concentrated ultrafine particulate matter in central Los Angeles. Mice were euthanized 24h after the last exposure and brain, liver, heart, lung and spleen tissues were collected and frozen for subsequent bioassays. There was clear evidence of aberrant immune activation in the brains of exposed animals as judged by a dose-related increase in nuclear translocation of two key transcription factors, NF-kappaB and AP-1. These factors are involved in the promotion of inflammation. Increased levels of glial fibrillary acidic protein (GFAP) were also found consequent to particulate inhalation suggesting that glial activation was taking place. In order to determine the mechanism by which these events occurred, levels of several MAP kinases involved in activation of these transcription factors were assayed by Western blotting. There were no significant changes in the proportion of active (phosphorylated) forms of ERK-1, IkB and p38. However, the fraction of JNK in the active form was significantly increased in animals receiving the lower concentration of concentrated ambient particles (CAPs). This suggests that the signaling pathway by which these transcription factors are activated involves the activation of JNK. PMID- 18420362 TI - Pharmacological basis for use of Pistacia integerrima leaves in hyperuricemia and gout. AB - ETHNOPHARMACOLOGICAL SIGNIFICANCE: Pistacia integerrima Stew ex. Brandis is an important component of commonly dispensed traditional dosage forms. We wished to determine whether polyphenolic constituents of this plant could be useful in oxidative stress and have potential to counter hyperuricemia. MATERIAL AND METHODS: Radical scavenging activity was determined by 1,1-diphenyl-2 picrylhydrazyl (DPPH) and xanthine oxidase (XO) inhibitory activity assay in vitro. Fructose (FRS) induced hyperuricemic animal model was used to asses the serum uric acid (UA) lowering effect by plant products. RESULTS: Ethyl acetate and n-BuOH fractions had the highest DPPH radical scavenging activity. Fifty percent inhibitory concentration (IC(50)) was 6 and 7.6 microg/ml respectively. It was less than quercetin (IC(50) 0.95 microg/ml) and ascorbic acid (IC(50) 1.76 microg/ml). Xanthine oxidase inhibitory activity was comparable between n-BuOH and EtOAc (IC(50) 19 and 20 microg/ml) extracts but less than quercetin (IC(50) 0.65 microg/ml) and allopurinol (IC(50) 0.10 microg/ml). The antioxidant activity as well as the inhibitory activity towards the enzyme XO by quercetin-3-O-beta-d glucopyranoside (5), kaempferol-3-O-beta-d-glucopyranoside (6), quercetin-3-O (6''-O-syringyl)-beta-d-glucopyranoside (7), kaempferol-3-O-(4''-O-galloyl)-alpha l-arabinopyranoside (8), rutin (4) together with aglycons, quercetin (1), kaempferol (2) and apigenin (3) was promising to continue in vivo hypouricemic studies. Ethyl acetate extract had dose dependent UA lowering effect in hyperuricemic mice. This effect was comparable with quercetin but less than allopurinol. CONCLUSIONS: These findings are encouraging to plan clinical studies in hyperuricemic patients. PMID- 18420363 TI - Diuretic activity of some Withania aristata Ait. fractions. AB - We previously reported on the significant dose-dependent diuretic effects produced in laboratory rats by hot water infusions and methanol extracts of Withania aristata Ait., where notable increases were observed in the excretion of water and sodium, with an interesting potassium-saving effect. The present study gives the results of the diuretic effects in rats of the hexane, dichloromethane, ethyl acetate, butanol and methanol-water fractions of the previously studied methanol extract. Water excretion rate, pH, density, conductivity and content of Na(+), K(+) and Cl(-) were measured in the urine of the rats when subjected to hypersaline conditions. Of the above fractions, the methanol:water extract (100mg/kg) showed the most interesting diuretic activity (25%; p<0.01), which suggested that increase in the polarity of the extracting solvent led to an increase in the concentration of the polar compounds responsible for the diuretic effect. These data, together with previous results on the aqueous and methanol extracts, reaffirm assertions made regarding the effectiveness of the extracts of this plant against urinary pathologies in the Canary Islands folk medicine. PMID- 18420364 TI - Characterization of human DNA in environmental samples. AB - Environmental samples from indoor surfaces can be confounded by dust, which is composed largely of human skin cells and has been documented to contain roughly tens of micrograms of total DNA per gram of dust. This study complements previous published work by providing estimates of the quantity of amplifiable human DNA found in environmental samples from a typical indoor environment, categorized by the intensity of human traffic and visible quantity of dust. Dust was collected by surface swabbing standard 576 cm(2) areas in eight locations, and evaluated for total DNA quantity, presence of human DNA (mitochondrial and nuclear loci using conventional PCR), quantity of human nuclear DNA using quantitative PCR, and STR analysis. The total DNA content of 36 dust samples ranged from 9 to 28 ng/cm(2), and contained 0.2-1.1 pg/cm(2) of human DNA. Overall, human DNA was detected in 97% of 36 dust samples and 61% of samples yielded allele distributions of varying degrees of complexity when subjected to STR analysis. The implications of this study are twofold. First, the presence of dust in evidence can be a significant contamination source in forensic investigations because the human DNA component is of sufficient quality and quantity to produce allele calls in STR analysis. This can be effectively managed by implementing stringent protocols for collection and analysis of potential biological samples. A second implication is the use of dust as a source of evidence for identification of inhabitants within a defined location. In the latter case, a number of additional studies would be necessary to identify relevant pretreatments for environmental dust samples and to develop the necessary deconvolution techniques to separate the composite genotypes obtained. PMID- 18420365 TI - Foreword for shoulder imaging volume of EJR. PMID- 18420366 TI - Diffusion anisotropy indexes are sensitive to selecting the EPI readout-encoding bandwidth at high-field MRI. AB - Diffusion tensor magnetic resonance imaging (DT-MRI) is generally performed using an echo planar imaging (EPI) acquisition to map directional water diffusion. However, the oscillating magnetic field gradients of the EPI acquisition can result in considerable mechanical vibrations, which lead, in turn, to magnetic field fluctuations causing Nyquist ghosting in the EPI data. The objective of this study was to investigate effects of EPI readout gradient modulation frequency, which is directly associated with the EPI readout bandwidth (BW), on the accuracy of DT-MRI measurements in a high magnetic field system. A spherical water phantom was used to study the relationship between the EPI BW and the Nyquist ghost for a spin-echo EPI acquisition with a matrix size of 128x128, complemented by diffusion sensitization gradients of up to b=800 s/mm(2) along six directions for DT-MRI. Nine volunteers (four males and five females) were studied using EPI at different BW acquisitions. Analysis of variance was used to investigate the EPI BW effects. The phantom studies demonstrated a systematic relationship between BWs and the intensities of Nyquist ghosts. In the human brain studies, EPI BW variations substantially corrupted diffusion anisotropy indexes (i.e., fractional anisotropy and relative anisotropy) (F=10.5, P=.0001) but were unrelated to diffusion-encoding directions (F=0.14, P=.98). It was possible to minimize BW dependence (F=1.48, P=.25) by tuning the modulation frequency of the EPI readout gradient. In conclusion, diffusion anisotropic indexes are sensitive to the readout BW of EPI due to associated Nyquist ghosting. However, the effect can be minimized by tuning the modulation frequency of the EPI readout gradient, that is, the EPI BW, to a range outside the harmonics of mechanical gradient vibrations. PMID- 18420367 TI - Real-time color-flow MRI at 3 T using variable-density spiral phase contrast. AB - The purpose of this study was to demonstrate and evaluate the performance of real time color-flow MRI at 3 T using variable-density spiral (VDS) phase contrast. Spiral phase contrast imaging was implemented within a flexible real-time interactive MRI system that provided continuous image reconstruction and an intuitive user interface. The pulse sequence consisted of a spectral-spatial excitation, bipolar gradient, spiral readout and gradient spoiler. VDSs were utilized to increase spatial and/or temporal resolution relative to uniform density spirals (UDSs). Parameter choices were guided by specific applications. Sliding window reconstruction was used to achieve a maximum display rate of 40 frames/s. No breath-holding or gating was used. Our results demonstrated that real-time color-flow movies using UDS and VDS provided adequate visualization of intracardiac flow, carotid flow and proximal coronary flow in healthy volunteers. Average aortic outflow velocity measured at the aortic valve plane using VDS was 29.4% higher than that using UDS. Peak velocity measured in the common carotid artery using VDS was 9.8% higher than that using UDS. PMID- 18420368 TI - MRI assessment of drug-induced fluid accumulation in humans: validation of the technology. AB - PURPOSE: The purpose of this study was to evaluate the feasibility of using proton and sodium magnetic resonance imaging (MRI) to detect fluid accumulation produced by fludrocortisone and nifedipine - two drugs known to cause salt/water retention by different mechanisms. MATERIALS AND METHODS: Twelve young healthy male subjects were randomly assigned to one of two groups and treated with either fludrocortisone or nifedipine for 14 or 25 days, respectively. The change in sodium MRI, as well as in proton T(2) value and T(1)-weighted signal intensity in the calf following postural change [referred to here as 'postural delta signal'(PDS)], was evaluated before, during and after drug administration. The changes in MRI PDS were compared to conventional physiological parameters, including body weight, calf volume and pitting edema. RESULTS: When compared to the baseline pretreatment values, the subjects treated with fludrocortisone showed a 5.5% increase in sodium MRI PDS (P=.01), a 2-ms increase in proton T(2) PDS of the gastrocnemius muscle (P=.06) and a body weight gain of 2.3% (P=.001) within 1 week. In the nifedipine-treated subjects, the sodium MRI PDS increased by 6% versus baseline (P=.03), while the proton T(2) PDS of the gastrocnemius muscle increased by 3.7 ms (P=.01), associated with a 0.5% weight gain (P=.55), within 3 weeks. No significant changes were noted in the T(1)-weighed images following postural change. Measurements of calf circumference, volume and pitting edema did not show consistent changes associated with the drug administration. CONCLUSION: The postural change in sodium MRI and proton T(2) signals provides a sensitive method for detecting the fluid accumulation produced by fludrocortisone and nifedipine. The MRI results are consistent with treatment-induced increases in extracellular fluid volume and correlate well with the observed weight gain. These findings support the potential utility of MRI for the evaluation of medication-induced fluid retention. PMID- 18420369 TI - Liquid chromatography-tandem mass spectrometry assay for the quantitation of plagiochin E and its main metabolite plagiochin E glucuronides in rat plasma. AB - Plagiochin E, a macrocyclic bisbibenzyl isolated from liverwort Marchantia polymorpha, was found to have antifungal activity. To evaluate the pharmacokinetics of plagiochin E in rats, a sensitive and specific liquid chromatography/tandem mass spectrometric (LC-MS/MS) method was developed and validated for the quantitation of plagiochin E and its total conjugated metabolites in rat plasma. For detection, a Sciex API 4000 LC-MS/MS with a TurboIonSpray ionization (ESI) inlet in the negative ion-multiple reaction monitoring (MRM) mode was used. The plasma samples were pretreated by a simple liquid-liquid extraction with ethyl acetate. The concentration of plagiochin E parent form was determined directly and the concentration of plagiochin E conjugated metabolites was assayed in the form of plagiochin E after treatment with beta-glucuronidase/sulfatase. The statistical evaluation for this method reveals excellent linearity, accuracy and precision for the range of concentrations 0.5-1000.0 ng/mL. The method had a lower limit of quantification (LLOQ) of 0.5 ng/mL for plagiochin E in 50 microL of plasma. The method was successfully applied to the characterization of the pharmacokinetic profile of plagiochin E in rats after an oral and an intravenous administration. PMID- 18420370 TI - Diagnostic performance of a new automated heterophile antibody test in adults and children. AB - Epstein-Barr virus serologic profiles were obtained from 618 patients using an automated platform, including the new BioPlex heterophile assay. When compared with 2 IgM viral capsid antigen (VCA) assays and a predicate heterophile assay, the BioPlex heterophile assay was more likely to be associated with serologic patterns of acute infection. PMID- 18420371 TI - [Haemostasis disorders caused by snake venoms]. AB - Various haemostasis disorders can occur following snakebite. Almost all ophidian species that are responsible for serious, even lethal, human envenomations are concerned. Venoms of these snakes are rich in proteins interfering with haemostasis, including many enzymes. These proteins can be classified in four groups according to their action. The haemorrhagins induce capillary permeability disorders. The proteins disturbing the primary haemostasis can activate as well as inhibit platelets: phospholipases A2, serine proteases and metalloproteinases, L-amino-acido-oxydases, phosphoesterases, disintegrins, C-type lectins, dendropeptin, agregoserpentin, thrombolectin. The proteins interfering with coagulation are separated into procoagulant proteases (prothrombin activator, thrombin-like enzymes, factor X and factor V activators) and anticoagulant proteases (factor IX and X inhibitors, protein C activator, anticoagulant phospholipases A2). The venom components acting on fibrinolysis are the fibrinolytic enzymes and the plasminogene activators. The clinical consequence of these mechanisms is a local as well as diffuse haemorrhagic syndrome. A hypofibrinogenemy, even an afibrinogenemy is frequently noted. Other haemostasis parameters are disturbed: PT collapse, a patient's ACT several times higher than the control and non-systematic thrombopenia. Ophidian venoms take part in many medical, diagnostic or therapeutic, applications in medicine. Currently, the antivenomous immunotherapy is the only efficient treatment in these haemorrhagic disorders. PMID- 18420372 TI - [Cardiac disease of glycogen-storage disease type II]. PMID- 18420373 TI - The ability of reaction time tests to detect simulation: an investigation of contextual effects and criterion scores. AB - Two experiments examined whether experience gained with a series of reaction time tests [Computerized Tests of Information Processing (CTIP); Tombaugh, T. N. & Rees, L. (in press). Computerized Tests of Information Processing (CTIP). Toronto, Canada: Multi-Health Systems Inc.] influenced the performance of individuals instructed to simulate the cognitive effects of a traumatic brain injury. Experience with the tests was manipulated by varying the order and number of tests administered for simulator and control groups. Simulators responded significantly slower and exhibited increased variability compared to controls. Performance was not affected by order or number of tests. The results of a third experiment showed that criterion scores could be established that correctly classified members of control, simulator, mild TBI, and severe TBI groups. Overall, the results suggest that the performance of the simulators was based on a context-free, absolute judgment and that reaction time measures show considerable promise for detecting low effort. PMID- 18420374 TI - Interpretation bias and social anxiety in adolescents. AB - Interpretation bias, described as the tendency to interpret social situations in a negative or threatening manner, has been widely linked to social anxiety in adult populations. This study aimed to extend research on interpretation bias to an adolescent population. Thirty-seven high socially anxious and a control group of 36 non-socially anxious adolescents rated the likelihood of different interpretations of ambiguous social and non-social situations coming to mind and which interpretation they most believed. Results showed that negative interpretations of social situations were more common in the high anxious than control group. Such negative bias could not be accounted for by high levels of negative affect. The groups did not differ as to their positive interpretations. Furthermore, there was evidence for content specificity of interpretation bias; high anxious adolescents were not more negative than control participants in their interpretations of non-social situations. Findings are discussed in relation to the adult literature and their clinical relevance is considered. PMID- 18420375 TI - Ecophysiological significance of scale-dependent patterns in prokaryotic genomes unveiled by a combination of statistic and genometric analyses. AB - We combined genometric (DNA walks) and statistical (detrended fluctuation analysis) methods on 456 prokaryotic chromosomes from 309 different bacterial and archaeal species to look for specific patterns and long-range correlations along the genome and relate them to ecological lifestyles. The position of each nucleotide along the complete genome sequence was plotted on an orthogonal plane (DNA landscape), and fluctuation analysis applied to the DNA walk series showed a long-range correlation in contrast to the lack of correlation for artificially generated genomes. Different features in the DNA landscapes among genomes from different ecological and metabolic groups of prokaryotes appeared with the combined analysis. Transition from hyperthermophilic to psychrophilic environments could have been related to more complex structural adaptations in microbial genomes, whereas for other environmental factors such as pH and salinity this effect would have been smaller. Prokaryotes with domain-specific metabolisms, such as photoautotrophy in Bacteria and methanogenesis in Archaea, showed consistent differences in genome correlation structure. Overall, we show that, beyond the relative proportion of nucleotides, correlation properties derived from their sequential position within the genome hide relevant phylogenetic and ecological information. This can be studied by combining genometric and statistical physics methods, leading to a reduction of genome complexity to a few useful descriptors. PMID- 18420377 TI - Autistic children: a neuroimmune perspective. PMID- 18420376 TI - Hematopoietic response of rats exposed to the impact of an acute psychophysiological stressor on responsiveness to an in vivo challenge with Listeria monocytogenes: modulation by Chlorella vulgaris prophylactic treatment. AB - In this study, we investigated the hematopoietic response of rats pretreated with CV and exposed to the impact of acute escapable, inescapable or psychogenical stress on responsiveness to an in vivo challenge with Listeria monocytogenes. No consistent changes were observed after exposure to escapable footshock. Conversely, the impact of uncontrollable stress (inescapable and psychogenical) was manifested by an early onset and increased severity and duration of myelossuppression produced by the infection. Small size CFU-GM colonies and increased numbers of clusters were observed, concurrently to a greater expansion in the more mature population of bone marrow granulocytes. No differences were observed between the responses of both uncontrollable stress regimens. CV prevented the myelossuppression caused by stress/infection due to increased numbers of CFU-GM in the bone marrow. Colonies of cells tightly packed, with a very condensed nucleus; in association with a greater expansion in the more immature population of bone marrow granulocytes were observed. Investigation of the production of colony-stimulating factors revealed increased colony stimulating activity (CSA) in the serum of normal and infected/stressed rats treated with the algae. CV treatment restored/enhanced the changes produced by stress/infection in total and differential bone marrow and peripheral cells counts. Further studies demonstrated that INF-gamma is significantly reduced, whereas IL-10 is significantly increased after exposure to uncontrollable stress. Treatment with CV significantly increased INF-gamma levels and diminished the levels of IL-10. Uncontrollable stress reduced the protection afforded by CV to a lethal dose of L. monocytogenes, with survival rates being reduced from (50%) in infected rats to 20% in infected/stressed rats. All together, our results suggest Chlorella treatment as an effective tool for the prophylaxis of post-stress myelossupression, including the detrimental effect of stress on the course and outcome of infections. PMID- 18420378 TI - The development and evaluation of a nested PCR assay for detection of Neospora caninum and Hammondia heydorni in feral mouse tissues. AB - The development of a novel nested polymerase chain reaction is described and used for detecting the presence of Neospora caninum and Hammondia heydorni DNA in DNA extracted from feral rodent tissues. A unique strategy was used for design of an assay that could be adapted for detecting DNA from more than one member of Toxoplasmatinae simultaneously with a minimal number of additional steps. The level of sensitivity described for this assay is comparable to real time-PCR and other nested PCR assays. Twenty-eight of 104 feral mice tested positive for N. caninum in at least one tissue (the brain, heart or liver) studied. In this study, eight instances are reported where the brain tested negative to N. caninum while at least one other tissue was positive. This suggests that prior studies, which screened only the brain, describe prevalence levels that are under represented. None of 54 mouse brains tested positive for H. heydorni DNA. This suggests that mice are rarely infected by H. heydorni although this hypothesis needs to be explored further. Data obtained in the current study suggest that N. caninum is a common parasite of feral rodents which may be important in the epidemiology of the disease. PMID- 18420379 TI - Mapping of CGRP in the alpaca (Lama pacos) brainstem. AB - In this study, we demonstrate the presence of immunoreactive structures containing calcitonin gene-related peptide in the alpaca brainstem. This is the first time that a detailed mapping of the cell bodies and fibers containing this neuropeptide in the alpaca brainstem has been carried out using an immunocytochemical technique. Immunoreactive cell bodies and fibers were widely distributed throughout the alpaca brainstem. A high density of calcitonin gene related peptide-immunoreactive perikarya was found in the superior colliculus, the dorsal nucleus of the raphe, the trochlear nucleus, the lateral division of the marginal nucleus of the brachium conjunctivum, the motor trigeminal nucleus, the facial nucleus, the pons reticular formation, the retrofacial nucleus, the rostral hypoglossal nucleus, and in the motor dorsal nucleus of the vagus, whereas a high density of fibers containing calcitonin gene-related peptide was observed in the lateral division of the marginal nucleus of the brachium conjunctivum, the parvocellular division of the alaminar spinal trigeminal nucleus, the external cuneate nucleus, the nucleus of the solitary tract, the laminar spinal trigeminal nucleus, and in the area postrema. This widespread distribution indicates that the neuropeptide studied might be involved in multiple functions in the alpaca brainstem. PMID- 18420380 TI - Toluene-induced hearing loss in acivicin-treated rats. AB - Toluene can be considered an ototoxic chemical compound in the rat. Outer hair cells are particularly sensitive to this aromatic organic solvent or to one of its metabolites. The objective of the present study was to evaluate the possible role played by cysteine S-conjugates in the ototoxic process in Long-Evans rats. To this end, renal and hepatic metabolism of toluene was modified by treatment with acivicin, an inhibitor of gamma-glutamyl transferase (gamma-GT). First, the efficacy of the acivicin treatment was established from a dose-response investigation in which urinary gamma-GT was measured daily in rats exposed to 1750 ppm toluene, 6 h per day for five days. A twice weekly 5 mg/kg dose was reduced urinary gamma-GT by 70-78%. In a subacute experiment, rats were exposed to 1750 ppm toluene for four consecutive weeks, in which the efficacy of the acivicin treatment was monitored by quantifying the urinary end product of the conjugate pathway: benzyl mercapturic acid (BMA). A 38.5% decrease in BMA was measured at the end of the exposure period. Hearing impairment was evaluated using auditory (inferior colliculus) evoked potentials and completed with conventional histological approaches. The toluene-exposed and the acivicin treated rats exposed to toluene both had a 7-dB permanent auditory threshold shift at 16-20 kHz. Hair cell loss was not dependent on acivicin treatment. Therefore, the partial inhibition of gamma-GT did not modify the toluene ototoxicity, suggesting that toluene-induced hearing loss is not strongly mediated by the production of cysteine S-conjugates. However, the data do not rule out the possibility that these metabolites may play a minor role. PMID- 18420381 TI - Learning to play Go using recursive neural networks. AB - Go is an ancient board game that poses unique opportunities and challenges for artificial intelligence. Currently, there are no computer Go programs that can play at the level of a good human player. However, the emergence of large repositories of games is opening the door for new machine learning approaches to address this challenge. Here we develop a machine learning approach to Go, and related board games, focusing primarily on the problem of learning a good evaluation function in a scalable way. Scalability is essential at multiple levels, from the library of local tactical patterns, to the integration of patterns across the board, to the size of the board itself. The system we propose is capable of automatically learning the propensity of local patterns from a library of games. Propensity and other local tactical information are fed into recursive neural networks, derived from a probabilistic Bayesian network architecture. The recursive neural networks in turn integrate local information across the board in all four cardinal directions and produce local outputs that represent local territory ownership probabilities. The aggregation of these probabilities provides an effective strategic evaluation function that is an estimate of the expected area at the end, or at various other stages, of the game. Local area targets for training can be derived from datasets of games played by human players. In this approach, while requiring a learning time proportional to N(4), skills learned on a board of size N(2) can easily be transferred to boards of other sizes. A system trained using only 9 x 9 amateur game data performs surprisingly well on a test set derived from 19 x 19 professional game data. Possible directions for further improvements are briefly discussed. PMID- 18420382 TI - Oxidative stress and calpain inhibition induce alpha B-crystallin phosphorylation via p38-MAPK and calcium signalling pathways in H9c2 cells. AB - We investigated the response of alphaB-crystallin to oxidative stress and calpain inhibition in an attempt to elucidate the signalling pathways mediating its phosphorylation. Given the high expression levels of alphaB-crystallin in cardiac muscle one can evaluate the significance of its participation in preservation of homeostasis under adverse conditions. H9c2 cardiac myoblasts were used as our experimental model since their response reflects the signal transduction pathways activated by stress conditions in the myocardium. Thus, in H9c2 cells treated with H2O2 the mechanism regulating alphaB-crystallin phosphorylation was found to involve p38-MAPK/MSK1 as well as intracellular free calcium levels. Our immunocytochemical experiments demonstrated phosphorylated alphaB-crystallin to be co-localized with tubulin, potentially preserving cytoskeletal architecture under these interventions. In H9c2 cells treated with calpain inhibitors (ALLN, ALLM) alphaB-crystallin exhibited a p38-MAPK- and [Ca 2+](i)-dependent phosphorylation pattern since the latter was ablated in the presence of the selective p38-MAPK inhibitor SB203580 and calcium chelator BAPTA-AM. Calpain activity repression ultimately led to apoptosis confirmed by PARP fragmentation and chromatin condensation. However, the apoptotic pathway activated by ALLM and ALLN differed, underlying the diverse transduction mechanisms stimulated. In addition to this, an anti-apoptotic role for phospho-alphaB-crystallin was verified by confirmation of its interaction with pro-caspase 3, hindering its cleavage and subsequent activation. Collectively, our findings underline alphaB crystallin crucial role as a participant of cardiac cells early response to stressful stimuli compromising their survival. PMID- 18420383 TI - Objective measures of prospective memory do not correlate with subjective complaints in schizophrenia. AB - While a number of studies have shown that individuals with schizophrenia are impaired on various types of prospective memory, few studies have examined the relationship between subjective and objective measures of this construct in this clinical group. The purpose of the current study was to explore the relationship between computer-based prospective memory tasks and the corresponding subjective complaints in patients with schizophrenia, individuals with schizotypal personality features, and healthy volunteers. The findings showed that patients with schizophrenia demonstrated significantly poorer performance in all domains of memory function except visual memory than individuals with schizotypal personality disorder and healthy controls. More importantly, there was a significant interaction effect of prospective memory type and group. Although patients with schizophrenia were found to show significantly poorer performance on computer-based measures of prospective memory than controls, their level of subjective complaint was not found to be significantly higher. While subjective complaints of prospective memory were found to associate significantly with self reported executive dysfunctions, significant relationships were not found between these complaints and performance on a computer-based task of prospective memory and other objective measures of memory. Taken together, these findings suggest that subjective and objective measures of prospective memory are two distinct domains that might need to be assessed and addressed separately. PMID- 18420384 TI - K6PC-5, a sphingosine kinase activator, induces anti-aging effects in intrinsically aged skin through intracellular Ca2+ signaling. AB - BACKGROUND: Sphingosine-1-phosphate (S1P), a bioactive sphingolipid metabolite, regulates multiple cellular responses such as Ca(2+) signaling, growth, survival, and differentiation. Because sphingosine kinase (SK) is the enzyme directly responsible for the production of S1P, many factors have been identified that regulate its activity and subsequent S1P levels. To date, there are no reports to demonstrate a chemically induced, direct activation of SK. OBJECTIVE: Here we have studied the effects of K6PC-5 as a newly synthesized SK activator on fibroblast proliferation in both human fibroblasts and aged mouse skin. To demonstrate that K6PC-5 has S1P-mediated action mechanism in fibroblasts, we have measured SK-dependent intracellular Ca(2+) signaling. METHODS: Fibroblasts were cultured primarily from human foreskin and were used to study the effect of K6PC 5 and S1P on intracellular Ca(2+) signaling and fibroblast proliferation. Changes in intracellular Ca(2+) were detected by fluorescence with fura-2/AM. To study skin anti-aging effects of K6PC-5, we used intrinsically aged hairless mice (56 weeks old). RESULTS: K6PC-5 promoted fibroblast proliferation and procollagen production in human fibroblasts significantly. K6PC-5 induced intracellular Ca(2+) concentration ([Ca(2+)](i)) oscillations in human fibroblasts. Both dimethylsphingosine and dihydroxysphingosine, SK inhibitors, and the transfection of SK1-siRNA blocked the K6PC-5-induced increases in [Ca(2+)](i), an effect independent of the classical PLC/IP(3)-mediated pathway. The K6PC-5-induced [Ca(2+)](i) oscillations were dependent on thapsigargin-sensitive Ca(2+) stores and Ca(2+) entry. Topical application of K6PC-5 for 2 weeks to intrinsically aged hairless mice enhanced fibroblast proliferation, collagen production, and eventually increased dermal thickness (10%). K6PC-5 also promoted specific epidermal differentiation marker proteins, including involucrin, loricrin, filaggrin, and keratin 5, without any alterations on epidermal barrier function. CONCLUSION: These results suggest that K6PC-5 acts to regulate fibroblast proliferation through intracellular S1P production, and can further promote keratinocyte differentiation. We anticipate that the regulation of S1P levels may represent a novel approach for the treatment of skin disorders, including skin aging. PMID- 18420385 TI - Sequence analysis of filaggrin gene by novel shotgun method in Japanese atopic dermatitis. AB - BACKGROUND: Recent reports indicated that nonsense mutations in filaggrin (FLG) found in ichthyosis vulgaris (IV) patients are predisposing factors for atopic dermatitis (AD) with asthma. The exon 3 of FLG contains tandemly repeated, highly homologous, 11-13 sequence units of 972 or 975 bp, each of which corresponds to the coding sequence of the processed filaggrin with slight sequence difference. This unique gene structure has hampered the precise DNA sequence determination. OBJECTIVE: We developed a novel DNA sequencing method "FLG-shotgun" to directly characterize the mutations in Japanese AD patients. METHODS: We examined 24 Japanese AD patients with "FLG-shotgun" method. RESULTS: Multiple units of FLG were amplified by PCR using several sets of common primers for the conserved regions, and DNA sequences of each cloned PCR product were determined. Multiple reads of DNA sequences in both alleles were aligned and re-constructed to cover the entire coding regions. We found three major genotypes (A, B, and C) which represent different numbers (11-13) of homologous sequence units. Furthermore, we found two novel nonsense mutations; one mutation 8666-8667CC>GA on the unit 9 of allele B that causes a nonsense mutation S2899X in two patients and the other mutation 9887C>A on the unit 10 of allele B that causes a nonsense mutation S3296X in two patients. CONCLUSION: We found two novel FLG mutations by directly analyzing Japanese patients with AD. FLG-shotgun will provide a valuable tool to further define the nature of the AD phenotype associated with FLG mutations. PMID- 18420386 TI - Vitiligo onset removes congenital nevocellular nevus cells. PMID- 18420387 TI - Combined NMR and computational study for azide binding to human manganese superoxide dismutase. AB - Human manganese superoxide dismutase (MnSOD) labeled with 3-fluorotyrosine (Tyf) was complexed with the (15)N-labeled inhibitor azide ([(15)N(3)(-)]). The sample was characterized by solid-state NMR (SSNMR) spectroscopy ((19)F-MAS and (15)N CPMAS). Employing (19)F-(15)N-REDOR spectroscopy, we determined the distances between the fluorine label in Tyrosine-34 and the three (15)N-nuclei of the azide and the relative orientation of the azide in the binding pocket of the MnSOD. A distance of R(1)=4.85A between the (19)F-label of Tyf34 and the nearest (15)N of the azide and an azide-fluorotyrosine Tyf34 angle of 90 degrees were determined. These geometry data are employed as input for molecular modeling of the location of the inhibitor in the active site of the enzyme. In the computations, several possible binding geometries of the azide near the Mn-complex were assumed. Only when the azide replaces the water ligand at the Mn-complex we obtained a geometry of the azide-Mn-complex, which is consistent with the present NMR data. This indicates that the water molecule ligating to the Mn-complex is removed and the azide is placed at this position. As a consequence the azide forms an H bond with Gln143 instead with Tyf34, in contrast to non-(19)F-labeled MnSOD, where the azide is hydrogen bonded to the hydroxy group of Tyr34. PMID- 18420388 TI - MC3T3-E1 cell adhesion to hydroxyapatite with adsorbed bone sialoprotein, bone osteopontin, and bovine serum albumin. AB - Native bone tissue is composed of a complex matrix of collagen, non-collagenous proteins, and hydroxyapatite (HAP). Bone sialoprotein (BSP) and bone osteopontin (OPN) are members of the non-collagenous protein family termed the SIBLING (small integrin-binding ligand, N-linked glycoproteins) proteins, which are primarily found in mineralized tissues. Previously, OPN was shown to exhibit a preferential orientation for MC3T3-E1 cell adhesion when it was specifically bound to collagen, while the MC3T3-E1 cell adhesion was shown to be dependant on the conformational flexibility of BSP specifically bound to collagen. Additionally, OPN was shown to play a greater role than BSP for cell binding to collagen. In this work, the orientations and conformations of BSP and OPN specifically bound to HAP are probed under similar conditions. Radiolabeled adsorption isotherms were obtained for BSP and OPN on HAP formed from a simulated body fluid, and the results show that HAP has the capacity to bind significantly more BSP than OPN. An in vitro MC3T3-E1 cell adhesion assay was then performed to compare the cell binding ability of adsorbed BSP and OPN specifically bound to HAP. It was found that there is a preference for cell binding to HAP with adsorbed BSP as compared to OPN, but not to a statistically significant level. However, the maximum cell binding was observed on HAP substrates with adsorbed heat denatured bovine serum albumin (BSA). The influence of BSA on cell binding was shown to be concentration dependant and it is believed that the adsorbed BSA modulates the proliferation state of the bound cells. PMID- 18420389 TI - Characterization of alkanoyl-10-O-minocyclines in micellar dispersions as potential agents for treatment of human neurodegenerative disorders. AB - Minocycline is a widely used antibacterial agent. Moreover, it is also demonstrated to be effective in several neurodegenerative disorders, due to its antioxidant and anti-inflammatory activities. However, the last activity is only apparent at very high doses. In fact, minocycline poorly crosses the blood-brain barrier (BBB) due to its low lipophilicity and half-life. The present work details the physicochemical characterization of a series of alkanoyl-10-O minocycline derivatives (2-6), which are able to produce self-assembled aggregates in aqueous solution. The n-octanol/aqueous phase lipophilicity of minocycline and its derivatives were assessed by theoretical calculation, by shake-flask method, and by reversed-phase HPLC. Moreover, we determined their affinity for membrane phospholipids measuring their HPLC retention on phospholipid-based stationary phases, the so-called "Immobilized Artificial Membranes" (IAMs). Our results indicate high lipophilicity values for the minocycline derivatives (compounds 2-6); these values and the corresponding phospholipid affinities increase with the length of the hydrocarbon moiety substituent. Furthermore, the ability of the investigated alkanoyl-10-O minocycline derivatives to self-assemble could allow a direct administration by oral and intraperitoneal routes as supramolecular systems. The advantages are an enhancement of drug solubilization, a sustained release, and the consequent less frequent drug administration. Moreover, we can hypothesize the potential solubilization in the micellar core of other poorly water soluble drugs which could improve the therapeutic effects of the pharmaceutical formulation in a combined therapy. Given the high lipophilicity of the title derivatives, they can be supposed to offer higher half-life and a better BBB penetration than minocycline. Since the new derivatives retain the structural features related to the antioxidant and anti-inflammatory effects of minocycline, they can be regarded not only as long-acting antimicrobial agents but also as candidate drugs for a targeted treatment of mental illness. PMID- 18420390 TI - Comparative interaction of 2-hydroxypropyl-beta-cyclodextrin and sulfobutylether beta-cyclodextrin with itraconazole: phase-solubility behavior and stabilization of supersaturated drug solutions. AB - Cyclodextrins can increase the apparent solubility and dissolution rate of poorly water-soluble drug candidates improving their biopharmaceutical performance. The current data assess the ability of hydrophilic cyclodextrins to solubilize compounds via stabilization of supersaturated drug solutions presumably by inhibition of nucleation and arresting crystal growth. To these points, the effects of 2-hydroxypropyl-beta-cyclodextrin (HPbetaCD) and sulfobutylether-beta cyclodextrin (SBEbetaCD) on equilibrium solubility was assessed via phase solubility analysis as were the interactions of these excipients on drug solubility under conditions favoring supersaturation. Phase-solubility analysis indicated that different profiles were generated as a function of the cyclodextrin examined and the pH of the complexing medium. When kinetic solubility measurements were completed, the cyclodextrins were found to stabilize concentrations of itraconazole significantly in excess of their equilibrium solubility when supersaturated solutions were formed using the co-solvent/solvent quench approach. These solutions were stable over 240 min falling in concentration at the 24 h time point of the experiment unlike those formed using surfactants and other polymers which demonstrated a rapid decrease in concentration over time. These data suggest that hydrophilic cyclodextrins might be useful formulation adjuncts in supersaturating drug delivery systems. PMID- 18420391 TI - Age-related changes in the incidence of pineal gland calcification in Turkey: A prospective multicenter CT study. AB - The goal of this cross-sectional observational study was to determine the incidence of pineal gland calcification (PGC), to investigate the interaction of PGC and aging, and to compare the incidence of PGC among the populations living in Turkey. In a prospective study the rate of PGC on CT scans of 1376 individuals in six referral centers from different regions of Turkey was investigated, with emphasis on effects of climatological parameters and aging on PGC. It was found that the incidence of PGC increased rapidly after first decade and the increase remains gradual thereafter, higher in males than in females for all age groups. There was a significant difference for incidence and degree of PGC between different clinics and between both sexes (p<0.001). In addition, there was a significant difference for the degree of PGC between the clinics in low altitude group and those in high altitude group (p<0.001 for each). Logistic regression analysis revealed that age, sex, altitude and intensity of sunlight exposure significantly affected the risk of PGC (odds ratios (OR) 1.335, 95% confidence intervals (CI) 1.261-1.414, p<0.001; OR 1.900, 95% CI 1.486-2.428, p<0.001; OR 0.715, 95% CI 0.517-0.990, p<0.05; OR 0.997, 95% CI 0.994-0.999, p<0.01, respectively). Furthermore, by multiple linear regression analysis, high altitude and increased intensity of sunlight exposure were found to affect the degree of PGC (beta=0.003, p<0.001). It is concluded that there is a close relationship between PGC and the aforementioned parameters, supporting a link between the development of PGC and these. This study provides some reference data for new clinical studies on the putative role of pineal gland in future. PMID- 18420392 TI - Identification of gene transcript signatures predictive for estrogen receptor and lymph node status using a stepwise forward selection artificial neural network modelling approach. AB - OBJECTIVE: The advent of microarrays has attracted considerable interest from biologists due to the potential for high throughput analysis of hundreds of thousands of gene transcripts. Subsequent analysis of the data may identify specific features which correspond to characteristics of interest within the population, for example, analysis of gene expression profiles in cancer patients to identify molecular signatures corresponding with prognostic outcome. These high throughput technologies have resulted in an unprecedented rate of data generation, often of high complexity, highlighting the need for novel data analysis methodologies that will cope with data of this nature. METHODS: Stepwise methods using artificial neural networks (ANNs) have been developed to identify an optimal subset of predictive gene transcripts from highly dimensional microarray data. Here these methods have been applied to a gene microarray dataset to identify and validate gene signatures corresponding with estrogen receptor and lymph node status in breast cancer. RESULTS: Many gene transcripts were identified whose expression could differentiate patients to very high accuracies based upon firstly whether they were positive or negative for estrogen receptor, and secondly whether metastasis to the axillary lymph node had occurred. A number of these genes had been previously reported to have a role in cancer. Significantly fewer genes were used compared to other previous studies. The models using the optimal gene subsets were internally validated using an extensive random sample cross-validation procedure and externally validated using a follow up dataset from a different cohort of patients on a newer array chip containing the same and additional probe sets. Here, the models retained high accuracies, emphasising the potential power of this approach in analysing complex systems. These findings show how the proposed method allows for the rapid analysis and subsequent detailed interrogation of gene expression signatures to provide a further understanding of the underlying molecular mechanisms that could be important in determining novel prognostic markers associated with cancer. PMID- 18420393 TI - Primary cryotherapy with salvage external beam radiotherapy for locally recurrent prostate cancer. PMID- 18420394 TI - Prevalence of overt metastases in locally advanced breast cancer. AB - AIMS: Locally advanced breast cancer (LABC) represents a wide spectrum of clinical presentations and poses significant clinical challenges for both patients and their physicians. Before starting neoadjuvant therapy, most patients undergo staging investigations, including a radioisotope bone scan, liver ultrasound and chest X-ray as per our provincial guidelines. The aim of this study was to document the prevalence of metastatic disease using standard baseline radiological staging in patients with LABC. MATERIALS AND METHODS: A retrospective chart review was carried out for LABC patients at two large Canadian centres between 2003 and 2006. Data on tumour characteristics and baseline staging tests were collected. Information on any confirmatory imaging (bone X-ray, computed tomography, magnetic resonance imaging, positron emission tomography) undertaken due to the presence of suspicious baseline tests or due to worrying symptoms was also obtained. The prevalence of metastatic disease after each baseline imaging technique was analysed, as was the frequency of discordance between baseline staging tests and confirmatory imaging where applicable. RESULTS: In total, 144 patients with LABC were assessed. After initial staging investigations, 15 patients (10.4%) were diagnosed as having overt metastatic disease. Confirmatory imaging was carried out on 19 patients (13.2%); five (3.5%) for unexplained symptoms and 14 (9.7%) due to equivocal baseline imaging. These additional investigations isolated a further four subjects with metastatic disease, bringing the overall prevalence of overt metastases to 13.2%. CONCLUSIONS: Given that the rate of systemic relapse in patients with LABC is very high, current baseline staging investigations probably underestimate the true incidence of metastases. This study has shown that further confirmatory imaging can be helpful, especially in symptomatic patients, as it seems that negative baseline tests in these patients can be falsely reassuring. Accurate staging of patients with LABC has many advantages and, therefore, further research is needed to define the role of newer imaging modalities. PMID- 18420395 TI - Influence of n-3 polyunsaturated fatty acids on soluble cellular adhesion molecules as biomarkers of cardiovascular risk in young healthy subjects. AB - BACKGROUND AND AIM: Serum levels of soluble cellular adhesion molecules (CAMs) and blood lipid parameters have been used as markers of inflammatory processes associated with cardiovascular disease (CVD) events. The present study evaluated the effects of the intake of n-3 polyunsaturated fatty acids (PUFAs) in fish and fish oil within energy-restricted diets, on soluble intercellular adhesion molecule-1 (sICAM-1) and soluble vascular adhesion molecule-1 (sVCAM-1). METHODS AND RESULTS: Two hundred and seventy-five healthy European subjects aged between 20 and 40 years, were randomized to one of four hypocaloric dietary groups: control (sunflower oil capsules, no seafood), lean fish (3 x 150 g portions of cod/week), fatty fish (3 x 150 g portions of salmon/week), fish oil ((docosahexaenoic acid (DHA)+eicosapentaenoic acid (EPA) capsules, no seafood)). Diets rich in lean fish significantly decreased ICAM-1 levels, around 5% from baseline to endpoint (p<0.05), and had no effect on VCAM-1 levels. No significant differences were observed in sICAM-1 levels after the intervention with fatty fish or fish oils. On the other hand, these two seafood based diets were responsible for a significant increase of VCAM-1 levels [fatty fish; 16.1% and fish oil; 21.9%] respectively (p<0.05). CONCLUSIONS: CAMs as inflammatory biomarkers in young and healthy subjects are not conclusive for the evaluation of CVD risk. Hypocaloric fish diets had a different effect on CAMs, being lean fish responsible for the highest decrease in ICAM-1. On the other hand, VCAM-1 results allow speculation that a low dose of n-3 PUFA may be anti-inflammatory contrarily to a high dose which can have a pro-inflammatory effect. CAMs mechanism is complex and affected by multiple factors such as lifestyle, gender, and n-3 dose and source. PMID- 18420396 TI - Thermodynamic behavior of glassy state of structurally related compounds. AB - Thermodynamic properties of amorphous pharmaceutical forms are responsible for enhanced solubility as well as poor physical stability. The present study was designed to investigate the differences in thermodynamic parameters arising out of disparate molecular structures and associations for four structurally related pharmaceutical compounds--celecoxib, valdecoxib, rofecoxib, and etoricoxib. Conventional and modulated temperature differential scanning calorimetry were employed to study glass forming ability and thermodynamic behavior of the glassy state of model compounds. Glass transition temperature of four glassy compounds was in a close range of 327.6-331.8 K, however, other thermodynamic parameters varied considerably. Kauzmann temperature, strength parameter and fragility parameter showed rofecoxib glass to be most fragile of the four compounds. Glass forming ability of the compounds fared similar in the critical cooling rate experiments, suggesting that different factors were determining the glass forming ability and subsequent behavior of the compounds in glassy state. A comprehensive understanding of such thermodynamic facets of amorphous form would help in rationalizing the approaches towards development of stable glassy pharmaceuticals. PMID- 18420397 TI - A quantitative anatomical study on posterior mandibular interradicular safe zones for miniscrew implantation in the beagle. AB - With the increasing expansion of miniscrew anchorage use in orthodontic treatment, more and more studies have been and will be carried out on the biochemistry, biomechanics and side effects of miniscrews in vivo. In such studies, a beagle has been the most commonly used animal model and its mandibular interradicular zones have been the greatest focus of interest. However, interradicular miniscrews risk failure by being loosened due to collision with adjacent roots. Therefore, it is necessary for the surgeon to be familiar with the anatomy of a beagle's mandible, especially that of the interradicular zones. This study has been performed to investigate the beagle's mandibular interradicular safe zones for miniscrew implantation to provide an anatomical guide for this type of study. Twenty-four beagle corpses were collected. Their mandible specimens were ground parallel to the respective buccal alveolar surface using a model trimmer until a horizontal plane was obtained, which was then sectioned on the line passing each tooth's central groove. In the image of this plane, cut at 2, 4, 6, 8 and 10 mm beneath the top of the alveolar crest, the mesiodistal width between the roots of P2 and P3, P3 and P4, P4 and M1 and the mesial and distal roots of M1 were measured, respectively. Zones of mesiodistal width measurement larger than 3.2 mm were found between P4 and M1, below the 8mm cut and between the mesial and distal roots of M1, below the 4 mm cut. In addition, between P2 and P3, below the 8mm cut and between P3 and P4, below the 10 mm cut, the mesiodistal width measurement was larger than 2.2 mm. The mandibular interradicular safe zones for miniscrew implantation in the dog were located between the mesial and distal roots of M1 and between the roots of P4 and M1, where there was enough mesiodistal width. Alveolar bone was relatively narrow between P2 and P3, P3 and P4, where care must be taken during implanting. PMID- 18420398 TI - Hypocholesterolemic effect of stilbenes containing extract-fraction from Cajanus cajan L. on diet-induced hypercholesterolemia in mice. AB - Cajanus cajan (L) is a natural plant which contains a lot of potential active components. In the present study, we identified the effects of the stilbenes containing extract-fraction from Cajanus cajan L (sECC) on diet-induced (for 4 weeks) hypercholesterolemia in Kunming mice. All experimental mice were divided into 5 groups: control group, model group, sECC-treated with 200 or 100 mg/kg/day, and simvastatin group. The effects of sECC were investigated by monitoring serum and liver lipid profile (cholesterol homeostasis and triglyceride) as well as serum superoxide dismutase activity in those mice. To further explore the mechanism of sECC, hepatic 3-hydroxy-3-methylglutaryl-CoA reductase (HMG-CoA reductase), cholesterol 7alpha-hydroxylase (CYP7A1), and low density lipoprotein receptor (LDL receptor) expressions in cholesterol homeostasis were analyzed by reverse transcription PCR. After 4 weeks pretreatment, compared with model group, the increased serum and hepatic total cholesterol were markedly attenuated by sECC (200 mg/kg) by 31.4% and 22.7% (p<0.01), respectively, the triglyceride levels of serum and liver were also lowered by 22.98% and 14.39%, respectively. At the same time, serum LDL cholesterol decreased by 52.8% (p<0.01) accompanied with the activities of serum superoxide dismutase increased by 20.98%. Atherogenic index and body weight were also reduced markedly. The mRNA expressions of HMG-CoA reductase, CYP7A1, and LDL receptor were significantly enhanced in the mice administered with sECC (200 mg/kg/day), whereas those expressions were suppressed by the hypercholesterolemic diet. These data indicate that sECC reduces the atherogenic properties of dietary cholesterol in mice. Its hypocholesterolemic effect may involve enhancement of the hepatic LDL-receptor and cholesterol 7alpha-hydroxylase expression levels and bile acid synthesis. PMID- 18420399 TI - Acute inhibition of superoxide formation and Rac1 activation by nitric oxide and iloprost in human vascular smooth muscle cells in response to the thromboxane A2 analogue, U46619. AB - BACKGROUND: The over-production of superoxide (O(2)(-)) derived from NADPH oxidase (NOX) plays a central role in cardiovascular diseases. By contrast, nitric oxide (NO) and prostacyclin (PGI(2)) are vasculoprotective. The effect of the NO donor, NONOate and iloprost on O(2)(-) formation, p47(phox) and Rac(1) activation in human vascular smooth muscle cells (hVSMCs) was investigated. METHODS: hVSMCs were incubated with 10nM thromboxane A(2) analogue, U46619 for 16h, and then with apocynin (a NOX inhibitor), NONOate or iloprost for 1h and O(2)(-) measured spectrophometrically. The role of cyclic AMP and cyclic GMP was examined by co-incubation of drugs with protein kinase (PK) A and G inhibitors listed above. Rac(1) was studied using pull-down assays. RESULTS: NONOate and iloprost inhibited O(2)(-) formation, acutely, effects blocked by inhibition of PKG and PKA, respectively. Rac(1) and p47(phox) activation and translocation to the plasma membrane was completely inhibited by NONOate and iloprost, effects again reversed by co-incubation with PKG or PKA inhibitors. CONCLUSIONS: NO and PGI(2) block the acute activity of NOX in hVSMCs via the cGMP-PKG axis (for NO) and by the cAMP-PKA axis (for iloprost) through inhibition of Rac(1) and p47(phox) translocation. These findings have implications in the pathophysiology and treatment of CVD. PMID- 18420400 TI - Gasification characteristics of MSW and an ANN prediction model. AB - Gasification characteristics make up the important parts of municipal solid waste (MSW) gasification and melting technology. These characteristics are closely related to the composition of MSW, which alters with climates and seasons. It is important to find a practical way to predict gasification characteristics. In this paper, five typical kinds of organic components (wood, paper, kitchen garbage, plastic, and textile) and three representative types of simulated MSW are gasified in a fluidized-bed at 400-800 degrees C with the equivalence ratio (ER) in the range of 0.2-0.6. The lower heating value (LHV) of gas, gasification products, and gas yield are reported. The results indicate that gasification characteristics are different from sample to sample. Based on the experimental data, an artificial neural networks (ANN) model is developed to predict gasification characteristics. The training and validating relative errors are within +/-15% and +/-20%, respectively, and predicting relative errors of an industrial sample are below +/-25%. This indicates that it is acceptable to predict gasification characteristics via ANN model. PMID- 18420401 TI - Effects of nomegestrol acetate administration on central and peripheral beta endorphin and allopregnanolone in ovx rats. AB - The aim of this study was to investigate the effects of nomegestrol acetate (NOMAc) on the central nervous system by analyzing the neurosteroid allopregnanolone and the opioid beta-endorphin (beta-endorphin). 104 Wistar female rats were used in this study; one group of fertile and one group of ovariectomized rats were used as control. The others were ovariectomized and they underwent a 2-week oral treatment of NOMAc (0.05, 0.1, 0.2, 0.5, 1mg/kg/day), alone or with 0.05 mg/kg/day of estradiol valerate (E2V). Allopregnanolone and beta-endorphin were assessed in different brain areas and in circulation. Ovariectomy decreased allopregnanolone anywhere except in the adrenal gland and E2V reversed the effects of ovariectomy. 0.5 and 1mg/kg/day of NOMAc increased allopregnanolone levels in hippocampus. Combined administration of 1mg/kg/day of NOMAc plus E2V induced a further increase of allopregnanolone levels in hippocampus, hypothalamus, and anterior pituitary. NOMAc (1mg/kg/day) decreased the adrenal content of allopregnanolone, both by itself and associated with E2V. NOMAc increased hippocampal and hypothalamic content of beta-endorphin at the highest doses, and it increased positively E2V action, at 1mg/kg/day, also in anterior pituitary and plasma. These findings reinforce the clinical data regarding the capability of NOMAc to modulate the pathways involved in mood and behaviour. In fact, due to the NOMAc action on hippocampus, hypothalamus, and anterior pituitary, our results highlight the selectivity of NOMAc on part of the limbic system and the anterior pituitary, regarding both allopregnanolone and beta-endorphin. PMID- 18420402 TI - Odor reduction rate in the confinement pig building by spraying various additives. AB - The objective of this on-site experiment was to evaluate and compare the effectiveness of currently utilized various additives, i.e. tap water, salt water, digested manure, microbial additive, soybean oil, artificial spice and essential oil, to reduce odor emissions from the confinement pig building. Odor reduction rates were evaluated with respect to sensual odor (odor concentration index, odor intensity and odor offensiveness) and odorous compounds (ammonia and sulfuric odorous compounds). Of the additives investigated in this study, salt water, artificial spice and essential oil had a positive effect on reducing odor generation. The effectiveness of salt water was only observed on ammonia, showing the reduction rates as a function of time (t=immediately, 1h, 3h, 5h, and 24h after spraying) were 0.1%, 20%, 36%, 11% and 0.2% as compared to initial level before spraying. The odor intensity and offensiveness were lessened by spraying artificial spice and essential oil of which maximum reduction rates ranged from 60% to 80%. Additionally, the essential oil had a significant effect on reducing sulfuric odorous compounds for 24h after spraying, which implicates that it functioned as not only a masking agent but also as an antimicrobial agent. PMID- 18420403 TI - Positive and negative aspects of soda/anthraquinone pulping of hardwoods. AB - The positive aspects of the non-sulfur soda/anthraquinone (SAQ) process are mostly tied to improved energy efficiency while lower pulp brightness after bleaching is its most significant drawback. A credible method that quantifies bleachability as well as an approach that solves the problem for SAQ pulps from hardwoods will be described. A straight line correlation (R2=0.904) was obtained between O2 kappa number and final light absorption coefficient (LAC) value after standardized OD0EpD1 bleaching of nine hardwood kraft pulps from three laboratories and one pulp mill. The bleachability of pulps from four different soda processes catalyzed by anthraquinone (AQ) and 2-methylanthraquinone (MAQ) was compared to that of conventional kraft pulps by comparing O2 kappa number decrease and final LAC values. It was observed that a mild hot water pre hydrolysis improved the bleachability of SAQ pulps to a level equal to that of kraft. PMID- 18420404 TI - Studies on lipid production by Rhodotorula glutinis fermentation using monosodium glutamate wastewater as culture medium. AB - Microbial lipid, as a raw material for biodiesel, can be produced by Rhodotorula glutinis with the monosodium glutamate (MSG) wastewater. The effect of adding glucose to MSG wastewater on lipid production was studied in this paper. Three different strategies, including initial addition, fed-batch addition and glucose feedback addition were attempted. The results show that addition of glucose was found favorable not only for cell growth but also for lipid synthesis. Of the three adding methods glucose feedback addition was the most effective one: about 25 g/L of biomass, 20% of lipid content and 45% of COD degradation were obtained respectively. And the components of the resulted lipid using different addition strategies were further studied. PMID- 18420405 TI - Discovery of trypanocidal thiosemicarbazone inhibitors of rhodesain and TbcatB. AB - Human African trypanosomiasis (HAT) is caused by the protozoan parasite Trypanosoma brucei. The cysteine proteases of T. brucei have been shown to be crucial for parasite replication and represent an attractive point for therapeutic intervention. Herein we describe the synthesis of a series of thiosemicarbazones and their activity against the trypanosomal cathepsins TbcatB and rhodesain, as well as human cathepsins L and B. The activity of these compounds was determined against cultured T. brucei, and specificity was assessed with a panel of four mammalian cell lines. PMID- 18420406 TI - Conformation-activity relationship on novel 4-pyridylmethylthio derivatives with antiangiogenic activity. AB - We found 4-pyridylmethylthio derivative 1 to be very effective in using antiangiogenesis activity to prevent proliferation of HUVECs (Human Umbilical Vein Endothelial Cells), which was induced by vascular endothelial growth factor (VEGF). Compound 1 was equally effective in inhibiting VEGF receptor2 tyrosine kinase (KDR, IC(50)=26nM). We deduced that the inhibition was the result of binding the catalytic domain of VEGF receptor2 tyrosine kinase in a similar fashion to both phthalazine derivative PTK787 2 and anthranylamide derivative AAL993 3. In this report, we will describe the conformational analyses, from ab initio MO calculation and X-ray crystallographic analyses, of compound 1 and the analogs, which include non-active 9, all in comparison with 2 and 3. The conformation-activity relationships suggest that a nonbonded intramolecular interaction between the sulfur and the carbonyl oxygen of 1 was very important in inhibiting KDR. PMID- 18420407 TI - Synthesis and evaluation of sulfamide-type indolizidines as glycosidase inhibitors. AB - A practical synthesis of reducing sulfamide-derived iminosugar glycomimetics related to the indolizidine glycosidase inhibitor family is reported. The polyhydroxylated bicyclic system was built from readily accessible hexofuranose derivatives through a synthetic scheme that involves 5,6-cyclic sulfamides. Further intramolecular nucleophilic addition of the sulfamide nitrogen atom to the masked aldehyde group of the monosaccharide in the open chain form afforded the target sugar mimics. By starting from d-glucose and d-mannose precursors, 2 aza-3,3-dioxo-3-thiaindolizidine derivatives with hydroxylation profiles that matched those of (+)-castanospermine and 6-epi-(+)-castanospermine were obtained. In vitro screening against a panel of glycosidases evidenced a high selectivity towards alpha-mannosidase. PMID- 18420408 TI - Orally efficacious thrombin inhibitors with cyanofluorophenylacetamide as the P2 motif. AB - 2-Cyano-6-fluorophenylacetamide was explored as a novel P2 scaffold in the design of thrombin inhibitors. Optimization around this structural motif culminated in 14, which is a potent thrombin inhibitor (K(i)=1.2nM) that exhibits robust efficacy in canine anticoagulation and thrombosis models upon oral administration. PMID- 18420409 TI - 2-Aminomethyl piperidines as novel urotensin-II receptor antagonists. AB - A series of 2-aminomethyl piperidines has been discovered as novel urotensin-II receptor antagonists. The synthesis, initial structure-activity relationships, and optimization of the initial hit that resulted in the identification of potent, cross-species active, and functional urotensin-II receptor antagonists such as 1a and 11a are described. PMID- 18420410 TI - Feeding problems and malnutrition in spinal muscular atrophy type II. AB - The aim of the study was to conduct a survey using a dedicated questionnaire to assess feeding difficulties and weight gain in a population of 122 Spinal Muscular Atrophy (SMA) type II patients, aged between 1 and 47 years. All the answers were entered in a database and were analysed subdividing the cohort into age groups (1-5, 6-10, 11-14, 15-19, 20-29, and 30-50 years). Six out of our 122 patients (5%), all younger than 11 years, had weights more than 2SD above the median for age matched controls, whilst 45 (37%) had weights less than 2SD below the median. Chewing difficulties were reported in 34 of the 122 patients (28%) and limitation in the ability to open the mouth in 36 (30%) and both were increasingly more frequent with age. Swallowing difficulties were reported in 30 patients (25%). The results of our survey suggest that a number of patients with SMA type II have limited jaw opening, and chewing and swallowing difficulties. Our findings raise a few issues concerning standards of care that should be implemented in the monitoring and management of feeding difficulties and weight gain. PMID- 18420411 TI - In memoriam: Professor Sir Howard Dalton, FRS (1944-2008). PMID- 18420412 TI - Life in the extreme: thermoacidophilic methanotrophy. AB - Aerobic methane-oxidizing bacteria (methanotrophs) have a key role in the global carbon cycle, converting methane to biomass and carbon dioxide. Although these bacteria have been isolated from many environments, until recently, it was not known if they survived, much less thrived in thermoacidic environments, that is, locations with pH values of approximately 1 and temperatures greater than 50 degrees C. Recently, three independent studies have isolated unusual methanotrophs from such extreme environments, expanding the known functional and phylogenetic diversity of methanotrophs. PMID- 18420413 TI - The impact of RNA structure on picornavirus IRES activity. AB - Internal ribosome entry site (IRES) elements consist of cis-acting regions that recruit the translation machinery to an internal position in the mRNA. The biological relevance of RNA structure-mediated mechanisms involved in internal ribosome recruitment is now emerging from the structural and functional analysis of viral IRES elements. However, because IRES elements found in genetically distant mRNAs seem to be organized in different RNA structures, the definition of the structural requirements for IRES activity is challenging and demands multidisciplinary approaches. This review discusses the latest reports that establish a relationship between RNA structure and IRES function in picornavirus genomes, the first RNAs described to contain these specialized regulatory elements. PMID- 18420414 TI - Lateral gene transfer challenges principles of microbial systematics. AB - Evolutionists strive to learn about the natural historical process that gave rise to various taxa, while also attempting to classify them efficiently and make generalizations about them. The quantitative importance of lateral gene transfer inferred from genomic data, although well acknowledged by microbiologists, is in conflict with the conceptual foundations of the traditional phylogenetic system erected to achieve these goals. To provide a true account of microbial evolution, we suggest developing an alternative conception of natural groups and introduce a new notion--the composite evolutionary unit. Furthermore, we argue that a comprehensive database containing overlapping taxonomical groups would constitute a step forward regarding the classification of microbes in the presence of lateral gene transfer. PMID- 18420415 TI - The influence of different tibial stem designs in load sharing and stability at the cement-bone interface in revision TKA. AB - Total Knee Arthroplasty (TKA) changes mechanical loading of the knee joint. Bone loss in the tibia is commonly encountered at the time of the revision TKA. Restoration of lost bone support and joint stability are the primary challenges in revision TKA. Normally, these defects are treated with non-living structures like metallic augments or bone grafts (autografts or allografts). Alone, neither of these structures can provide the initial support and stability for revision implants. In the latter, the use of intramedullary stems can provide the necessary load sharing and protect the remaining host bone and graft from excessive stress, increasing component stability. The purpose of this study was to evaluate comparatively load sharing (cortical rim, cancellous bone and stem) and stability at the cement-bone interface under the tibial tray induced by the use of cemented and press-fit tibial component stem extensions. Furthermore the study of the desirable option in cases where the bone defect is cavitary (cancellous bone defect contained by an intact cortical rim) or uncontained bone defect (bone loss involving the supporting cortical rim) was carried out. Because in vitro evaluation of these biomechanical parameters is difficult we used finite element (FE) models to overcome this. The biomechanical results suggest an identical behaviour in case of cavitary defects for both types of stems assessed. In the case of uncontained defect treated with bulk allografts the cemented stem may be a prudent clinical option. PMID- 18420416 TI - The release of calcium from the endoplasmic reticulum induced by amyloid-beta and prion peptides activates the mitochondrial apoptotic pathway. AB - In this study, we analyzed whether ER Ca2+ release, induced by amyloid-beta (Abeta) and prion (PrP) peptides activates the mitochondrial-mediated apoptotic pathway. In cortical neurons, addition of the synthetic Abeta1-40 or PrP106-126 peptides depletes ER Ca2+ content, leading to cytosolic Ca2+ overload. The Ca2+ released through ryanodine (RyR) and inositol 1,4,5-trisphosphate (IP3R) receptors was shown to be involved in the loss of mitochondrial membrane potential, Bax translocation to mitochondria and apoptotic death. Our data further demonstrate that Ca2+ released from the ER leads to the depletion of endogenous GSH levels and accumulation of reactive oxygen species, which were also involved in the depolarization of the mitochondrial membrane. These results illustrate that the early Abeta- and PrP -induced perturbation of ER Ca2+ homeostasis affects mitochondrial function, activating the mitochondrial-mediated apoptotic pathway and help to clarify the mechanism implicated in neuronal death that occurs in AD and PrD. PMID- 18420417 TI - Effects of shade on the development and sugar metabolism of coffee (Coffea arabica L.) fruits. AB - Coffee fruits grown in shade are characterized by larger bean size than those grown under full-sun conditions. The present study assessed the effects of shade on bean characteristics and sugar metabolism by analyzing tissue development, sugar contents, activities of sucrose metabolizing enzymes and expression of sucrose synthase-encoding genes in fruits of coffee (Coffea arabica L.) plants submitted to full-sun (FS) and shade (SH) conditions. Evolution of tissue fresh weights measured in fruits collected regularly from flowering to maturation indicated that this increase is due to greater development of the perisperm tissue in the shade. The effects of light regime on sucrose and reducing sugar (glucose and fructose) contents were studied in fresh and dry coffee beans. Shade led to a significant reduction in sucrose content and to an increase in reducing sugars. In pericarp and perisperm tissues, higher activities of sucrose synthase (EC 2.4.1.13) and sucrose-phosphate synthase (SPS: EC 2.4.1.14) were detected at maturation in the shade compared with full sun. These two enzymes also had higher peaks of activities in developing endosperm under shade than in full sun. It was also noted that shade modified the expression of SUS-encoding genes in coffee beans; CaSUS2 gene transcripts levels were higher in SH than in FS. As no sucrose increase accompanied these changes, this suggests that sucrose metabolism was redirected to other metabolic pathways that need to be identified. PMID- 18420418 TI - A comparison of fluorescence in two sympatric scorpion species. AB - In order to test the feasibility of scorpion fluorescence as an indicator of gender and/or species identity, a comparison of the fluorescence spectra between genders across two sympatric species of scorpions (Vaejovis confusus Stahnke 1940 and Paruroctonus shulovi Williams 1970) was conducted. Each spectrum was represented in a simple multivariate analysis by its peak wavelength and width at 90% of peak intensity. No difference between genders was found, but a statistically significant difference between species was detected. The potential of fluorescence as a basis for species recognition based on this difference is discussed in the context of what is known about scorpion vision and discriminatory abilities. PMID- 18420420 TI - Expression and purification of the D4 region of PLD1 and characterization of its interaction with PED-PEA15. AB - PLD's (Phospholipases D) are ubiquitously expressed proteins involved in many transphosphatidylation reactions. They have a bi-lobed structure composed by two similar domains which at their interface reconstitute the catalytic site through the association of the two conserved HxKx(4)Dx(6)GSxN motifs. PLD1 interacts with the small phosphoprotein PED-PEA15 by an unknown mechanism that, by enhancing PLD1 stability, apparently increases its enzymatic activity; the minimum interacting region of PLD1 was previously identified as spanning residues 712 1074 (D4 region). Since the D4/PED-PEA15 interaction has been claimed to be one of the multiple molecular events that can trigger type 2 diabetes, we purified the two recombinant proteins to study in vitro this binding by both ELISA and SPR techniques. Whilst PED-PEA15 was easily expressed and purified, expression of recombinant D4 was more problematic and only the fusion protein with Thioredoxin A and a six Histidine Tag (Trx-His(6)-D4) demonstrated sufficient stability for further characterization. We have found that Trx-His(6)-D4 is present as two different oligomeric forms, though only the monomeric variant is able to interact with PED-PEA15. All these findings may have important implications for both the mechanisms of phospholipase activity and PED-PEA15 regulative functions. PMID- 18420421 TI - Fatigue as a major predictor of quality of life in women with autoimmune liver disease: the case of primary biliary cirrhosis. AB - OBJECTIVES: Fatigue is a nearly universal symptom of many chronic diseases, yet it is often poorly understood and underappreciated as a factor in quality of life (QOL). Generally, clinicians have relied on subjective measures of fatigue, if they consider it at all. This study uses well-validated instruments to examine fatigue as a predictor of QOL in women with primary biliary cirrhosis (PBC), an autoimmune, chronic liver disease. METHODS: Eighty-one women with PBC completed a survey that included measures of fatigue (Fatigue Impact Scale) and QOL (SF-36) as well as demographic variables (age, education) and medical information (symptoms, stage of illness, time since diagnosis). QOL results for the sample were compared with those of a nationally normed U.S. population. Bivariate and multivariate analyses were conducted to identify contributors to variation in QOL. RESULTS: Compared with national norms, QOL for this PBC population was significantly impaired. When all variables with bivariate significance in relation to QOL were included in multivariate analyses, results showed fatigue to be the primary predictor of QOL, including all 8 QOL scales and the 2 summary scales. Regression results, dominated by fatigue, explain 25-59% of the variance in QOL. CONCLUSIONS: Fatigue has profound effects on every aspect of life for women with PBC-physical, social, emotional, and psychological. The results lead to recommendations for health care providers to assess fatigue in their patients with PBC and to take steps, where warranted, to mitigate its effects. PMID- 18420422 TI - Isolation, purification and characterization of beta-1,3-glucan binding protein from the plasma of marine mussel Perna viridis. AB - A beta-1,3-glucan binding protein (betaGBP) specific for laminarin (a beta-1,3 glucan) was detected for the first time in a mollusc, Perna viridis. betaGBP was isolated and purified from the plasma using laminarin precipitation and affinity chromatography on laminarin-Sepharose 6B, respectively. It agglutinated bakers yeast, bacteria, and erythrocytes and enhanced prophenoloxidase (proPO) activity of the plasma in a dose-dependent manner. The purified betaGBP appeared as a single band in native-PAGE and the purity was conformed by HPLC. The protein has a molecular weight estimate of 510kDa as determined by SDS-PAGE and in isoelectric focusing the purified betaGBP was focused as a single band at pI 5.3. betaGBP was found to possess inherent serine protease activity but lacked beta 1,3-glucanase activity and all these results suggest that plasma betaGBP of P. viridis functions as a recognition molecule for beta-1,3-glucan on the surface of microbial cell walls. This recognition and binding lead to the activation of the prophenoloxidase cascade mediated by the inherent serine protease activity of betaGBP. Presence of agglutinating activity and serine protease activity shows that betaGBP is a bifunctional protein. The findings are discussed in light of the importance of this protein in the innate immune response of P. viridis, and they implicate evolutionary link with similar proteins found in other invertebrates. PMID- 18420419 TI - Muscle-like nicotinic receptor accessory molecules in sensory hair cells of the inner ear. AB - Nothing is known about the regulation of nicotinic acetylcholine receptors (nAChRs) in hair cells of the inner ear. MuSK, rapsyn and RIC-3 are accessory molecules associated with muscle and brain nAChR function. We demonstrate that these accessory molecules are expressed in the inner ear raising the possibility of a muscle-like mechanism for clustering and assembly of nAChRs in hair cells. We focused our investigations on rapsyn and RIC-3. Rapsyn interacts with the cytoplasmic loop of nAChR alpha9 subunits but not nAChR alpha10 subunits. Although rapsyn and RIC-3 increase nAChR alpha9 expression, rapsyn plays a greater role in receptor clustering while RIC-3 is important for acetylcholine induced calcium responses. Our data suggest that RIC-3 facilitates receptor function, while rapsyn enhances receptor clustering at the cell surface. PMID- 18420423 TI - Oscillatory correlates of intentional updating in episodic memory. AB - The efficient use of our memory does not only require remembering encoded information, it also requires forgetting old out-of-date information. That such memory updating is part of our memory system is suggested by numerous behavioral studies. The physiological correlates of this process, however, still remain elusive. In this study we explore oscillatory correlates of memory updating as they occur in list-method directed forgetting. In this task, subjects are cued to forget a previously learned word list and to learn a new list of words instead. Such cuing typically leads to forgetting of the first list (List 1) and to memory enhancement of the second (List 2). Measuring EEGs during List-2 encoding, we identified two effects of the forget cue on oscillatory function: an increase in upper alpha power and a reduction in upper alpha phase coupling (11 to 13 Hz). Median-split analyses revealed that the two oscillatory effects were selectively related to the two behavioral effects. Whereas the increase in power was related to List-2 enhancement, the reduced phase coupling was related to List-1 forgetting. Our results point to separate neural origins of forgetting and enhancement and show that alpha oscillations play a critical role in intentional updating of episodic memory. PMID- 18420424 TI - The path to success in auditory spatial discrimination: electrical neuroimaging responses within the supratemporal plane predict performance outcome. AB - Auditory scene analysis requires the accurate encoding and comparison of the perceived spatial positions of sound sources. The electrophysiological correlates of auditory spatial discrimination and their relationship to performance accuracy were studied in humans by applying electrical neuroimaging analyses to auditory evoked potentials (AEPs) that were recorded during the completion of a near threshold S1-S2 paradigm within the right hemispace. Data were sorted as a function of performance accuracy, and AEP responses 75-117 ms after the presentation of the first sound differed topographically between trials leading to correct and incorrect spatial discrimination. Distributed source estimations revealed that this followed from significantly stronger activity within the left (i.e. contralateral) supratemporal plane (STP) and the left inferior parietal lobule prior to correct versus incorrect discrimination performance. Successful spatial discrimination thus depends on the activity of distinct configurations of active brain networks within the contralateral temporo-parietal cortex over a time period when the first sound position is being encoded. Furthermore, significant positive correlations were observed between performance accuracy and the intracranial activity estimated within the left STP. The efficacy of S1 processing within the STP is thus predictive of behavioral performance outcome during auditory spatial discrimination. Our data support a model wherein refinement of spatial representations occurs within the STP and that interactions with parietal structures allow for transformations into coordinate frames that are required for higher-order computations including absolute localization of sound sources. PMID- 18420426 TI - Generation of novel motor sequences: the neural correlates of musical improvisation. AB - While some motor behavior is instinctive and stereotyped or learned and re executed, much action is a spontaneous response to a novel set of environmental conditions. The neural correlates of both pre-learned and cued motor sequences have been previously studied, but novel motor behavior has thus far not been examined through brain imaging. In this paper, we report a study of musical improvisation in trained pianists with functional magnetic resonance imaging (fMRI), using improvisation as a case study of novel action generation. We demonstrate that both rhythmic (temporal) and melodic (ordinal) motor sequence creation modulate activity in a network of brain regions comprised of the dorsal premotor cortex, the rostral cingulate zone of the anterior cingulate cortex, and the inferior frontal gyrus. These findings are consistent with a role for the dorsal premotor cortex in movement coordination, the rostral cingulate zone in voluntary selection, and the inferior frontal gyrus in sequence generation. Thus, the invention of novel motor sequences in musical improvisation recruits a network of brain regions coordinated to generate possible sequences, select among them, and execute the decided-upon sequence. PMID- 18420425 TI - Coupling between somatosensory evoked potentials and hemodynamic response in the rat. AB - We studied the relationship between somatosensory evoked potentials (SEP) recorded with scalp electroencephalography (EEG) and hemoglobin responses recorded non-invasively with diffuse optical imaging (DOI) during parametrically varied electrical forepaw stimulation in rats. Using these macroscopic techniques we verified that the hemodynamic response is not linearly coupled to the somatosensory evoked potentials, and that a power or threshold law best describes the coupling between SEP and the hemoglobin response, in agreement with the results of most invasive studies. We decompose the SEP response in three components (P1, N1, and P2) to determine which best predicts the hemoglobin response. We found that N1 and P2 predict the hemoglobin response significantly better than P1 and the input stimuli (S). Previous electrophysiology studies reported in the literature show that P1 originates in layer IV directly from thalamocortical afferents, while N1 and P2 originate in layers I and II and reflect the majority of local cortico-cortical interactions. Our results suggest that the evoked hemoglobin response is driven by the cortical synaptic activity and not by direct thalamic input. The N1 and P2 components, and not P1, need to be considered to correctly interpret neurovascular coupling. PMID- 18420427 TI - Multimodal imaging of human early visual cortex by combining functional and molecular measurements with fMRI and PET. AB - Receptor distribution patterns of neurotransmitters and distinct functional fields of the human brain appear to be tightly connected with respect to their topological allocation along the cerebral cortex. There is, however, considerable lack of human data directly demonstrating this association in vivo. Here, we assessed the relationship between the distribution of the major inhibitory serotonergic neurotransmitter receptor, the 5-HT(1A) subtype, and the functional organization within early visual cortex defined by retinotopic mapping. The 5 HT(1A) receptor-binding potential was quantified by positron emission tomography (PET) using the highly selective and specific radioligand [carbonyl-(11)C]WAY 100635 in seven healthy subjects. The retinotopic maps and borders determined by functional magnetic resonance imaging (fMRI) were compared to the receptor distribution employing surface-based region of interest analysis in each of these subjects. We found a significant difference in receptor-binding potential in the functionally defined primary (V1) compared to secondary (V2) visual area, as V1 exhibits only 68% of receptor binding found in V2 in both hemispheres, which is consistent with postmortem data. Our in vivo findings clearly support prior assumptions of a link between receptor distribution and functional fields of the human cortex. PMID- 18420428 TI - Cortical mechanisms of retinal and extraretinal smooth pursuit eye movements to different target velocities. AB - Smooth pursuit eye movements (SPEM) are used to maintain focus upon moving targets. The generation of SPEM velocity is controlled by retinal information and extraretinal signals. Although there is a wealth of studies investigating retinal and extraretinal SPEM control, the main questions regarding the cortical mechanisms involved in the processing of SPEM to different stimulus velocities are still unresolved. We applied an innovative event-related fMRI-design by presenting target ramps at different velocities (5, 10, 15, 20 degrees/s) with both continuous target presentation and intervals of target blanking. The stimulus parameters were integrated into the statistical model and eye movements were registered to confirm SPEM performance. Our results clearly demonstrate that in humans the oculomotor network (V5, frontal and supplementary eye fields, lateral intraparietal area) is engaged in the processing of retinal and extraretinal SPEM velocity. Within this network neural activity increases with increasing target velocity. During extraretinal SPEM, additional engagement of the dorsolateral prefrontal cortex, angular gyrus, parahippocampal gyrus and superior temporal gyrus occurs. These regions encode cognitive functions such as memory, attention and monitoring. The activation of the inferior parietal cortex seems to be related to the interaction between velocity and blanking thereby underlining its relevance for task switching and sensorimotor transformation. PMID- 18420429 TI - Lamotrigine and catamenial epilepsy. AB - Catamenial epilepsy (CE) is characterized by epileptic seizures in the female occurring rhythmatically with the menstrual cycle. Hormonal mechanisms have been proposed as a cause of this epileptic form. Few reports about the efficacy of anti-epileptic drugs (AEDs) have been published. We studied prospectively women with CE who were treated with lamotrigine (LTG) for a period of 3 months in order to evaluate its efficacy, measuring the progesterone levels before and after LTG at the same time. LTG seemed to be efficacious in 66% of women, meaning the disappearance of seizures or reduction of 50% or more of the number of seizures. The reported side effects were few and mild, and the drug was well tolerated. Serum progesterone levels were found to rise during LTG treatment. PMID- 18420430 TI - Avocado/soybean unsaponifiables, ASU EXPANSCIENCE, are strictly different from the nutraceutical products claiming ASU appellation. PMID- 18420431 TI - A comparison of the clinical anesthetic efficacy of 4% articaine and 0.5% bupivacaine (both with 1:200,000 epinephrine) for lower third molar removal. AB - OBJECTIVE: This study compared the clinical efficacy of 4% articaine (A200) and 0.5% bupivacaine (B200), both with 1:200,000 epinephrine, for lower third molar removal. STUDY DESIGN: Fifty patients underwent removal of symmetrically positioned lower third molars, in 2 separate appointments, under local anesthesia either with A200 or B200, in a double-blind, randomized, and crossover manner. Time to onset, duration of postoperative analgesia, duration of anesthetic action on soft tissues, intraoperative bleeding, and hemodynamic parameters were evaluated. RESULTS: A statistically significant difference between the time to onset of A200 (1.66 +/- 0.13 minutes) and B200 (2.51 +/- 0.21 minutes) was found (P < .05). There was no statistically significant difference in the duration of analgesia, whether the patient was subjected to osteotomy or not, regardless of the local anesthetic used (3 to 4 hours; P > .05). However, when patients received B200 they experienced a statistically significant longer period of anesthesia on the soft tissues as compared with when they had received A200 (around 5 hours and 4 hours, respectively, P < .05). The surgeon's rating of intraoperative bleeding was considered very close to minimal for both anesthetics. In the surgeries with osteotomy, the comparison between A200 and B200 showed statistically significant differences in the diastolic (64 mm Hg and 68 mm Hg, respectively, P = .001) and mean arterial pressure (86 mm Hg and 89 mm Hg, respectively, P = .031) when data from all the surgical phases were pooled. Additionally, the mouth opening at the suture removal was statistically different for A200 and B200 solutions (91.90% +/- 3.00% and 88.57% +/- 2.38% of the preoperative measure, respectively) when surgeries required bone removal (P < .05). CONCLUSIONS: In comparison with 0.5% bupivacaine, 4% articaine (both with 1:200,000 epinephrine) provided a shorter time to onset and comparable hemostasis and postoperative pain control with a shorter duration of soft tissue anesthesia in lower third molar removal. PMID- 18420432 TI - G80S-linked ferroportin disease: the first clinical description in a Greek family. PMID- 18420433 TI - CreA mediates repression of the regulatory gene xlnR which controls the production of xylanolytic enzymes in Aspergillus nidulans. AB - The Aspergillus nidulans xlnR gene encodes a Zn(2)Cys(6) transcription activator necessary for the synthesis of the main xylanolytic enzymes, i.e. endo-xylanases X(22), X(24) and X(34), and beta-xilosidase XlnD. Expression of xlnR is not sufficient for induction of genes encoding the xylanolytic complex, the presence of xylose is absolutely required. It has been established previously that the wide-domain carbon catabolite repressor CreA indirectly represses xlnA (encodes X(22)) and xlnB (encodes X(24)) genes as well as exerting direct repression on xlnA. This work provides evidence that CreA-mediated indirect repression occurs through repression of xlnR: (i) the xlnR gene promoter is repressed by glucose and this repression is abolished in creA(d)30 mutant strains and (ii) deregulated expression of xlnR completely relieves glucose repression of xlnA and xlnB. Thus, CreA and XlnR form a transcriptional cascade regulating A. nidulans xylanolytic genes. PMID- 18420434 TI - Aspergillus nidulans ChiA is a glycosylphosphatidylinositol (GPI)-anchored chitinase specifically localized at polarized growth sites. AB - It is believed that chitinases play important physiological roles in filamentous fungi since chitin is one of the major cell wall components in these organisms. In this paper we investigated a chitinase gene, chiA, of Aspergillus nidulans and found that the gene product of chiA consists of a signal sequence, a region including chitinase consensus motifs, a Ser/Thr/Pro-rich region and a glycosylphosphatidylinositol (GPI)-anchor attachment motif. Phosphatidylinositol specific phospholipase C treatment of the fusion protein of ChiA and enhanced green fluorescent protein (EGFP)-ChiA-EGFP-caused a change in its hydrophobicity, indicating that ChiA is a GPI-anchored protein. ChiA-EGFP localized at the germ tubes of conidia, at hyphal branching sites and hyphal tips. chiA expression was specifically high during conidia germination and in the marginal growth regions of colonies. These results suggest that ChiA functions as a GPI-anchored chitinase at the sites where cell wall remodeling and/or cell wall maturation actively take place. PMID- 18420435 TI - Drop hand in a 12-year-old girl: radial nerve at peril due to knitting. AB - Reported here is a 12-year-old girl who suffered numbness and weakness in her right hand. She was diagnosed to have a right-sided radial nerve injury proximal to the supinator muscle. To the best knowledge of the authors, this is the first report of a child with radial nerve entrapment due to intense knitting. PMID- 18420436 TI - Variability in leucocyte profiles in thin-billed prions Pachyptila belcheri. AB - Because immune function competes for the resources that can be allocated to other activities, studies of immunological ecology may offer a powerful tool for explaining how reproductive effort links to reproductive costs and how conditions experienced early in their development affect growing chicks in later life. We studied the distribution of leucocyte types and the development of H/L ratio, which is indicative of heightened energetic stress, throughout the season 2004 2005 in chicks and adults of thin-billed prions Pachyptila belcheri. Adults decreased body condition throughout the season and increased H/L ratios. Likewise, chicks increased H/L ratios during the season, but this was age-related rather than condition-dependent. Chicks from earlier hatched eggs had lower H/L ratios initially, but this relationship became weaker with increasing age and had disappeared by fledging. The results suggest that the stress index may be a useful measure of condition in adult thin-billed prions, at least on a population level, although a larger sample size or repeated samples from the same individuals may be required to confirm the relationship on an individual level and to distinguish between seasonal and body condition effects. The data on chicks highlight our lack of knowledge of the ontogeny of immune function in wild birds. Studies of adults and chicks over several seasons may reveal how resources are allocated between immune and other functions under contrasting environmental conditions. PMID- 18420437 TI - Characterization of the hemoglobins of the neonatal brushtailed possum Trichosurus vulpecula (Kerr): evidence for a highly cooperative, aggregated isoform of hemoglobin. AB - The red blood cells of the neonatal brushtailed possum exhibit unusually strong cooperativity at high levels of oxygen saturation (n=5.4) which appear to arise from a concentration dependent aggregation of one of the neonatal hemoglobin isoforms. Red blood cells from neonatal pouched young exhibit a Bohr factor of 0.36. Stripped hemolysate is sensitive to added 2,3-bisphosphoglycerate (BPG) (apparent binding constant K=35 micromol L(-1)) and ATP (K=180 micromol L(-1)), but is largely insensitive towards chloride ions. Five isoforms of non-adult hemoglobin were identified using isoelectric focusing. Mass spectrometry indicated that two early isoforms contain alpha chains identical to the adult alpha chain. The remaining three isoforms are composed of identical alpha type and beta type gene products, but differ in their isoelectric points due to differential post-translational modification. PMID- 18420438 TI - Blood components: current challenges. AB - Labile blood products are still here to stay for quite a long time. All European surveys come in agreement to reveal that indications, extremely heterogeneous from one country to another, are not fully under control. Both the EuroNet-TMS network and Euro'SAT congresses aim at improving the use of such products within an evidence-based medicine perspective. PMID- 18420439 TI - Platelet additive solutions: a future perspective. AB - Platelet additive solutions (PASs) were first developed in the 1980s, and continued to be improved over the following years. The use of PASs as replacement for plasma has a number of benefits, both for the quality of the platelet concentrates and for the patients. However, some PASs have been associated with a lower platelet yield in the PCs, a shorter storage time, and a lower increment in the patient when compared to PCs in plasma. A number of reformulations of the PASs have taken place to counteract these disadvantages. Most PASs use acetate as nutrient for the platelets, which has the benefit of generating bicarbonate when oxidized by the platelets, thus supplying its own buffering capacity. Alternatively, glucose is used, but may cause deterioration of pH in the stored PCs due to the formation of lactic acid. Addition of other buffering substances, such as phosphate, can be added to ensure maintenance of neutral pH. An important finding was the inhibiting effect of potassium and magnesium on platelet activation. The initially developed PASs lacked these two ingredients and showed reduced storage times of the PCs in PAS when compared to those stored in plasma. However, when these constituents are included in the PAS, storage time is similar and even exceeds those seen for PCs in plasma. Considerable research is done in further formulating the optimal PAS. Bicarbonate is being considered as buffer for these PASs. Also, L-carnitine appears to have a favorable effect on stored platelets, including a reduction of platelet metabolism, and inhibition of apoptosis. Another area of optimization is lowering of plasma content needed for maintaining platelet quality. Where current PASs still need at least 30% residual plasma, there is a trend towards lowering the plasma content to less than 5% with the newer PASs. Preservation of purinergic platelet receptor functionality by ADP degrading activities in plasma appears to play an important role in this respect. Development of PASs are usually based on in vitro studies alone. It is important to realize that only clinical studies can give definitive answers about the quality of platelets stored in PASs. Sofar, only limited clinical evaluations have been published that either studied the effectiveness of platelets in initially-developed PASs, or were specifically done in combination with pathogen reduction technologies. Thus, PASs seem to be an excellent replacement for (part of) the plasma when producing PCs, and allow extended storage with maintenance of quality, but more clinical studies are needed to substantiate in vitro results. PMID- 18420440 TI - [Resectable adenocarcinoma of the oesophagogastric junction care: which perioperative treatment?]. AB - Adenocarcinoma of the oesophagogastric junction has an ominous prognosis. Until now, oesophageal adenocarcima care was close to the squamous cell cancer one whereas adenocarcinoma of the cardia was mixed with gastric cancers. Results from randomised studies mixed them without making distinctions. Nevertheless, context, natural history and clinical outcome differ. Five-year survival rate is around 40 %, all stages included. Results from several phase-III studies or meta-analysis allowed to define three therapeutic strategies applicable to adenocarcinoma of the oesophagus and the oesophagogastric junction. In Europe, in the case of a resectable tumour, preoperative chemotherapy became a standard treatment since results from the Magic trial. In the United States, post-operative radiochemotherapy according to the "Macdonald" scheme is used in case of a resected tumour with a R0 surgery. Actually, modern techniques of irradiation could reduce the rate of gastro-intestinal toxicities. The survival benefit from preoperative radiochemotherapy is still very controversial with high rates of postoperative morbidity and mortality. We have performed a review of the literature with a methodological analysis of data with a high level of evidence in order to advise perioperative treatment guidelines for patients with a resectable adenocarcinoma of the lower oesophagus or gastro-oesophageal junction. Results from pre- or postoperative strategies and the role of radiotherapy will need to be analysed in the future through a randomised study. PMID- 18420441 TI - [Is external cephalic version for breech presentation useful?]. PMID- 18420442 TI - [Trophoblastic pregnancy persistant disease with a menopausal woman: ultrasonography-Doppler benefits]. AB - We present the case of a 53-year-old menopaused woman who developed an invasive persistent trophoblastic uterine disease with several lung metastasis. Patient case was listed in a highly risked group in the WHO classification. As the patient refused primary hysterectomy she received polychemotherapy alone. Monitoring of the treatment and vascularity evolution of the tumor was followed through pelvic endovaginal ultrasound using 3D and contrast enhancement as well as HCG decrease in the context of uterus conservation that was wished by the patient. PMID- 18420443 TI - [How I perform... salvage of peritoneal haemorrhage in ruptured ectopic pregnancies in Benin]. PMID- 18420445 TI - [Maternal adrenal necrosis in the third trimester of pregnancy: a rare complication of antiphospholipid syndrome]. AB - Adrenal necrosis, a rare life threatening complication of antiphospholipid syndrome, is difficult to diagnose during pregnancy. We report the case of a 33 year-old woman with bilateral adrenal necrosis which started during the third trimester of her second pregnancy. Antiphospholipid syndrome had been diagnosed few years ago, after a thrombotic event. The pregnancy was uneventful until 36 weeks plus five days, when the patient was admitted for bilateral back ache, initially considered as uterine contractions. Labour was induced because pain persisted and was associated with major thrombocytopenia. A healthy infant was delivered vaginally on the second day, adrenal failure was diagnosed based on intense asthenia, persistent severe lumbar pain, low blood sodium and cortisol. Bilateral adrenal oedema was documented by CT scan and MRI. Symptoms resolved following administration of hydrocortisone and fludrocortisone. This case illustrates the difficulty to diagnose adrenal necrosis in the third trimester of pregnancy. PMID- 18420446 TI - Neighbourhoods and mental well-being: what are the pathways? AB - The last decade has seen an acceleration in research examining the impact of neighbourhood characteristics on a health outcomes. Yet, identifying specific pathways by which neighbourhoods affect mental well-being has received less attention. The article describes findings from a qualitative study conducted in Toronto, Ont., Canada, examining the pathways by which neighbourhood characteristics are related to mental well-being. Methods of concept mapping were used to answer our research questions. Participants identified over 100 neighbourhood characteristics grouped into six cluster that were important for either good or poor mental well-being. Clusters were rated in terms of their importance to mental well-being. Finally, participants drew diagrams describing how the neighbourhood factors and domains were contributing to poor and good mental well-being. Concept mapping provides unique contributions and challenges and may provide a useful means of examining specific pathways from neighbourhood to health outcomes. PMID- 18420447 TI - Coagulation signalling following tissue injury: focus on the role of factor Xa. AB - The primary function of the coagulation cascade is to promote haemostasis and limit blood loss in response to tissue injury. However, it is now recognized that the physiological functions of the coagulation cascade extend beyond blood coagulation and that this cascade plays a pivotal role in influencing inflammatory and tissue repair responses via the activation of their signalling responses, the proteinase-activated receptors (PARs). Consequently, uncontrolled coagulation activity and PAR signalling contributes to the pathophysiology of several conditions, including thrombosis, arthritis, cancer, kidney disease, and acute and chronic lung injury. Much of the work thus far has focused on the role of thrombin-mediated signalling in the pathophysiology of these conditions. However, recent evidence suggests that coagulation proteinases upstream of thrombin, including factor Xa (FXa), may also signal via PARs and thus induce cellular effects independent of thrombin generation. These studies have highlighted a novel and important role for FXa signalling in influencing proinflammatory and pro-fibrotic effects following tissue injury. This article will provide an overview of FXa as a central proteinase of the coagulation cascade and will review more recent evidence that FXa signalling may contribute to inflammation and tissue remodelling. The novel opportunities that this may present for therapeutic intervention will also be highlighted. PMID- 18420448 TI - Hierarchical models of behavior and prefrontal function. AB - The recognition of hierarchical structure in human behavior was one of the founding insights of the cognitive revolution. Despite decades of research, however, the computational mechanisms underlying hierarchically organized behavior are still not fully understood. Recent findings from behavioral and neuroscientific research have fueled a resurgence of interest in the problem, inspiring a new generation of computational models. In addition to developing some classic proposals, these models also break fresh ground, teasing apart different forms of hierarchical structure, placing a new focus on the issue of learning and addressing recent findings concerning the representation of behavioral hierarchies within the prefrontal cortex. In addition to offering explanations for some key aspects of behavior and functional neuroanatomy, the latest models also pose new questions for empirical research. PMID- 18420449 TI - Genotoxic effects of the pesticides Rubigan, Omite and Rovral in root-meristem cells of Crepis capillaris L. AB - Three pesticides have been studied for their genotoxicity by the use of assays in the plant Crepis capillaris, aimed at measuring chromosomal aberrations, micronuclei and sister chromosome exchange (SCE). The fungicides Rubigan 12 EC (fenarimol) and Rovral 25 Flo (iprodione) and the insecticide Omite 57 E (propargite) are all widely used nowadays. The aim of our study was to evaluate the genotoxic effects of these pesticides at concentrations corresponding to those applied in agricultural practice. In preliminary experiments we found that these concentrations do not influence cell proliferation and do not inhibit the growth of root meristems. In all experiments formulated commercial products were used. From the results we conclude that the three pesticides did not induce chromosomal aberrations as estimated by metaphase and anaphase analyses. They were also not capable to induce SCE. Rubigan did not induce micronucleus formation even at the highest concentration tested, but Omite and Rovral markedly increased micronucleus formation. The MN response depended on the sampling time and the concentration used, which showed a significant dose-response correlation (r=0.978, P<0.01 and r=0.941, P<0.01, respectively). A greater increase in micronucleus frequency was observed after Rovral treatment, where the highest concentration gave a response 8-10-fold above the negative control. Both pesticides induced high frequencies of lagging chromosomes, even after exposure to the lower test concentrations. The presence of lagging chromosomes is an indication of anti-microtubule activity of the pesticides tested. This effect was more strongly expressed after exposure to the two higher concentrations of Omite and Rovral. In this case a complete destruction of the mitotic spindle was observed, resulting in C-mitoses as well as in numerical aberrations-polyploidy and aneuploidy. The present findings suggest that Omite and Rovral at concentrations comparable to those used in practice can be regarded as potential aneugens. PMID- 18420450 TI - Genetic susceptibility to malignant pleural mesothelioma and other asbestos associated diseases. AB - Exposure to asbestos fibers is a major risk factor for malignant pleural mesothelioma (MPM), lung cancer, and other non-neoplastic conditions, such as asbestosis and pleural plaques. However, in the last decade many studies have shown that polymorphism in the genes involved in xenobiotic and oxidative metabolism or in DNA repair processes may play an important role in the etiology and pathogenesis of these diseases. To evaluate the association between diseases linked to asbestos and genetic variability we performed a review of studies on this topic included in the PubMed database. One hundred fifty-nine citations were retrieved; 24 of them met the inclusion criteria and were evaluated in the review. The most commonly studied GSTM1 polymorphism showed for all asbestos linked diseases an increased risk in association with the null genotype, possibly linked to its role in the conjugation of reactive oxygen species. Studies focused on GSTT1 null and SOD2 Ala16Val polymorphisms gave conflicting results, while promising results came from studies on alpha1-antitrypsin in asbestosis and MPO in lung cancer. Among genetic polymorphisms associated to the risk of MPM, the GSTM1 null genotype and two variant alleles of XRCC1 and XRCC3 showed increased risks in a subset of studies. Results for the NAT2 acetylator status, SOD2 polymorphism and EPHX activity were conflicting. Major limitations in the study design, including the small size of study groups, affected the reliability of these studies. Technical improvements such as the use of high-throughput techniques will help to identify molecular pathways regulated by candidate genes. PMID- 18420451 TI - A convenient method for the preparation of barbituric and thiobarbituric acid transition metal complexes. AB - A convenient method for the preparation of barbiturate transition metal complexes: (i) Cr(3+), Mn(2+), Fe(3+), Zn(2+) and Cd(2+) ions with barbituric acid (H(2)L) and (ii) Cr(3+) and Mo(5+) with 2-thiobarbituric acid (H(2)L') was reported and this has enabled seven complexes to be formulated as: [Cr(HL)(2)(OH)(H(2)O)].H(2)O, [Mn(HL)(2)(H(2)O)(2)], [Fe(2)(L)(OH)(3)(H(2)O)(4)].2H(2)O, [Zn(HL)(2)], [Cd(HL)(2)], [Cr(HL')(OH)(2)(H(2)O)].H(2)O and [Mo(HL')(2)]Cl. These new barbiturate complexes were synthesized and characterized by elemental analysis, molar conductivity, magnetic measurements, spectral methods (mid infrared, (1)H NMR, mass, X-ray powder diffraction and UV/vis spectra) and simultaneous thermal analysis (TG and DTG) techniques. The molar conductance measurements proved that, all complexes of barbituric and 2-thiobarbituric acids are non-electrolytes except for [Mo(HL')(2)]Cl. The electronic spectra and magnetic susceptibility measurements were used to infer the structures. The IR spectra of the ligands and their complexes are used to identify the mode of coordination. Kinetic and thermodynamic parameters such as: E, DeltaH, DeltaS and DeltaG are estimated according to the DTG curves. The two ligands and their complexes have been studied for their possible biological antifungal activity. PMID- 18420452 TI - Photoinduced interaction between riboflavin and TiO(2) colloid. AB - The adsorption of riboflavin on the surface of TiO(2) colloidal particles and the electron transfer process from its singlet excited state to the conduction band of TiO(2) were examined by absorption and fluorescence quenching measurements. The apparent association constants (K(app)) were determined. The quenching mechanism is discussed involving electron transfer from riboflavin to TiO(2). PMID- 18420453 TI - Study of the interactions of sodium thiocyanate, potassium thiocyanate and ammonium thiocyanate in water+N,N-dimethylformamide mixtures by Raman spectroscopy. AB - Raman spectra of water+N,N-dimethylformamide (DMF) mixtures and their solutions with NaNCS, KNCS and NH(4)NCS were obtained. The bands of nu(CO) stretching, delta(OCN) bending, r(CH(3)) rocking and nu(N-CH)(3)) stretching of the DMF molecule with and without salts were studied. The dependence of the vibration frequencies and Raman intensities of the bands on the composition of the mixed solvent was discussed. The change of the band frequencies as a result of the presence of the salts and the solvation of the cations by the solvent molecules was examined. The stronger cation solvation by the aprotic solvent molecules instead of the water molecules in DMF concentrated solutions was discussed. The nu(CN) and nu(CS) vibrations of the SCN(-) ions were observed as a function of the cation present and the solvent composition. The presence of the SCN(-) ions as "free", contact ion pairs, or solvent separated pairs, was discussed. PMID- 18420454 TI - Genetic drift of norovirus genotype GII-4 in seven consecutive epidemic seasons in Hungary. AB - BACKGROUND: Noroviruses are common pathogens in human gastroenteritis outbreaks worldwide. They belong to a genetically diverse group of RNA viruses with multiple genogroups (G) and genotypes. Genotype GII-4 norovirus (Lordsdale) is the predominant agent in epidemics. OBJECTIVES: To investigate the genetic variation in GII-4 strains isolated during seven epidemic seasons in Hungary from November 2000 to June 2007. STUDY DESIGN: Using the prospective epidemiological surveillance of norovirus outbreaks in Hungary, GII-4 strains were selected for further genetic analysis. After phylogenetic analysis, RNA-polymerase (open reading frame 1; ORF1), capsid (ORF2) and the ORF1/ORF2 junction were analysed by RT-PCR and sequencing. RESULTS: Three hundred and seventy-seven (76.8%) of 491 confirmed norovirus outbreaks were caused by genotype GII-4. GII-4 was the predominant genotype in six of the seven epidemic seasons. Four main GII-4 variants--epidemic point mutants--(GII-4-2000, GII-4-2002, GII-4-2004 and GII-4 2006b) were detected, with each variant predominating in two consecutive epidemic seasons. CONCLUSIONS: Genotype GII-4 was confirmed as the predominant genetic type in epidemic norovirus seasons in Hungary. Genetic drift successfully promotes the re-emergence of GII-4 variants in the population. The elevated number of norovirus outbreaks in the population predicts the emergence of new GII 4 genetic variants as part of an international epidemic. PMID- 18420457 TI - Molecular tools for the rapid detection of drug resistance in animal trypanosomes. AB - There are currently 17 African countries in which animal trypanocidal drug resistance has been reported. Large-scale surveys were carried out in only ten of them. The lack of baseline information is mainly due to the fact that the methods currently available for the detection of drug resistance are laborious, expensive and time consuming. In this review the mechanisms involved in resistance to isometamidium and diminazene will be discussed, together with some new molecular detection tools that have been developed recently enabling faster diagnosis of drug resistance than conventional laboratory or field tests. PMID- 18420458 TI - Hot, sweet and sticky: the glycobiology of Plasmodium falciparum. AB - Carbohydrate structures that decorate the surface of cells are increasingly recognized as playing important roles in the biology of host-pathogen interactions. Plasmodium species have undergone a process of gene loss that has removed much of their capacity to produce complex glycoconjugates or glycosylated proteins other than the glycosylphosphatidyinositol (GPI) moiety that anchors the surface proteins of infective stages, including the merozoite. Instead, these parasites have elaborated a set of proteins with lectin-like properties that interact with mammalian and insect cell surfaces. An overview of this and other aspects of the glycobiology of Plasmodium is presented here. PMID- 18420459 TI - Confronting an influenza pandemic with inexpensive generic agents: can it be done? AB - Avian influenza A H5N1 presents a serious and possibly imminent pandemic threat. In such an event, adequate supplies of affordable vaccines and antiviral agents will be unavailable to most people in the world. In view of the overwhelming need for effective alternatives, generic agents that target the host immune response or the pandemic virus should be considered. Many scientists doubt the effectiveness of these agents. Nonetheless, several studies suggest that statins improve outcomes in patients with bacteraemia and pneumonia and might be similarly effective against influenza. An experimental study has shown that the fibrate gemfibrozil, a peroxisome proliferator-activated receptor (PPAR) alpha agonist, reduces mortality in H2N2 influenza virus-infected mice. There is substantial molecular cross-talk between statins and PPAR agonists, and their clinical effects are additive in patients with cardiovascular diseases. Chloroquine increases endosomal pH, impairing influenza virus release into the cytosol. Statins, fibrates, and chloroquine are produced as generic medications in developing countries. They are inexpensive, could be stockpiled, and would be available on the first pandemic day. With a lack of realistic alternatives for confronting the next pandemic, research is urgently needed to determine whether these and other generic agents could mitigate the effects of what might otherwise become an unprecedented global public-health crisis. PMID- 18420460 TI - The influence of sex and ovarian hormones on temporomandibular joint nociception in rats. AB - The aim of this study was to investigate the influence of sex and ovarian hormones on formalin- and glutamate-induced temporomandibular joint (TMJ) nociception in rats. The influence of sex and ovarian hormones on the nociceptive behavior induced by formalin or glutamate was virtually the same. The nociceptive behavior of males was similar to that of females in the proestrus phase of the estrous cycle but was significantly lower than that in the diestrus phase. Since the serum level of estradiol but not of progesterone was significantly higher in the proestrus than in the diestrus phase, these data suggest that females with lower endogenous serum level of estradiol have an exacerbation of TMJ nociception. The nociceptive behavior of ovariectomized rats was similar to that of diestrus females and significantly greater than that of proestrus females. Although the administration of estradiol or progesterone in ovariectomized females significantly reduced TMJ nociception, the combination of both hormones did not increase the antinociceptive effect induced by each of them. These findings suggest that estradiol and progesterone decrease TMJ nociception in an independent way. PERSPECTIVE: We report that ovarian hormones have an antinociceptive effect on the TMJ formalin and glutamate nociceptive behavior models. Therefore, the greater prevalence and severity of TMJ pain in women of reproductive age may be a consequence of hormonal fluctuation during the reproductive cycle, in that during low endogenous estradiol serum level TMJ pain sensitivity is increased, enhancing the risk of females experiencing TMJ pain. PMID- 18420461 TI - Activation of the alpha7-nicotinic acetylcholine receptor reverses complete freund adjuvant-induced mechanical hyperalgesia in the rat via a central site of action. AB - The role of specific nicotinic receptor (nAChR) subtypes in antinociception has not been fully elucidated because of the lack, until recently, of selective tool compounds. (R)-N-(1-azabicyclo[2.2.2]oct-3-yl)(5-(2-pyridyl)thiopene-2 carboxamide) (compound B) is reported to be an agonist selective for the alpha(7)nAChR and in the present study was found to be efficacious in inflammatory pain models in 2 species. Compound B reversed complete Freund adjuvant-induced reductions in paw withdrawal thresholds in rat and mouse in a dose-related manner, producing maximum reversals of 65% +/- 4% at 10 mg/kg and 87% +/- 15% at 20 mg/kg. When rats and mice were predosed with the centrally penetrant, broad-spectrum nicotinic receptor antagonist mecamylamine, the efficacy of the agonist was significantly inhibited, producing reversals of only 11% +/- 5% at 10 mg/kg and 5% +/- 13% at 20 mg/kg, confirming activity via nicotinic receptors. Rats were also predosed systemically with the selective low brain penetrant alpha(7)-antagonist methyllycaconitine, which had no effect on agonist activity (90% +/- 18% at 10 mg/kg), suggesting a central involvement. This hypothesis was further established with methyllycaconitine completely inhibited the agonist effect when dosed intrathecally (1% +/- 7%). PERSPECTIVE: These studies provide good rationale for the utility of selective, central nervous system penetrant agonists at the alpha(7)-nicotinic receptor for the treatment of inflammatory pain. PMID- 18420462 TI - Influence of metal concentrations, percent salinity, and length of exposure on the metabolic rate of fathead minnows (Pimephales promelas). AB - Understanding the effects of chemical toxicants on energetic processes is an important aspect of ecotoxicology. However, the influence of toxicant concentration and time of exposure on metabolism in aquatic organisms is still poorly understood. The purpose of this investigation was to determine the influence of increasing levels of three stressors (Cu, Cd, percent salinity) and exposure time (24 h and 96 h) on the metabolic rate of fathead minnows (Pimephales promelas). In all 24-h exposures, there existed a threshold concentration, above which metabolic rate decreased significantly compared to the control and lower concentrations. In contrast, the metabolic rate of fish exposed for 96 h increased significantly in all concentrations compared to fish from the control. We suggest fathead minnows exhibit a consistent pattern of metabolic response to stressors, regardless of the physiological mechanisms involved, and that this response differs as a function of time of exposure. PMID- 18420463 TI - Theoretical considerations underlying Na(+) uptake mechanisms in freshwater fishes. AB - Ion and acid-base regulating mechanisms have been studied at the fish gill for almost a century. Original models proposed for Na(+) and Cl(-) uptake, and their linkage with H(+) and HCO(3)(-) secretion have changed substantially with the development of more sophisticated physiological techniques. At the freshwater fish gill, two dominant mechanisms for Na(+) uptake from dilute environments have persisted in the literature. The use of an apical Na(+)/H(+) exchanger driven by a basolateral Na(+)/K(+)-ATPase versus an apical Na(+) channel electrogenically coupled to an apical H(+)-ATPase have been the source of debate for a number of years. Advances in molecular biology have greatly enhanced our understanding of the basic ion transport mechanisms at the fish gill. However, it is imperative to ensure that thermodynamic principles are followed in the development of new models for gill ion transport. This review will focus on the recent molecular advances for Na(+) uptake in freshwater fish. Emphasis will be placed on thermodynamic constraints that prevent electroneutral apical NHE function in most freshwater environments. By combining recent advances in molecular and functional physiology of fish gills with thermodynamic considerations of ion transport, our knowledge in the field should continue to grow in a logical manner. PMID- 18420464 TI - Laparoscopic restoration of gastrointestinal continuity after duodenal switch. PMID- 18420465 TI - Alcohol abuse and dependence before and after bariatric surgery: a review of the literature and report of a new data set. AB - BACKGROUND: To review the published data on alcohol abuse and alcohol dependence after bariatric surgery and to assess the prevalence of alcohol abuse and alcohol dependence in a sample of bariatric surgery patients, both before and after surgery. All participant data were collected from patients who had undergone gastric bypass at a Midwestern hospital. METHODS: The participants were contacted by mail 6-10 years after surgery and asked to complete the Post-Bariatric Surgery Appearance Questionnaire. RESULTS: The results of the analysis indicated that a small percentage of the sample appeared to spontaneously develop alcohol abuse or dependence after bariatric surgery. CONCLUSION: The results of the present study will be useful in understanding the likelihood of bariatric surgery patients developing alcohol abuse and dependence after surgery. However, additional research is needed to fully understand the risk of this outcome. PMID- 18420466 TI - An in silico immunological approach for prediction of CD8+ T cell epitopes of Leishmania major proteins in susceptible BALB/c and resistant C57BL/6 murine models of infection. AB - It is well established that MHC class II restricted-CD4 T cells are dominant during the development of immunity against Leishmania (L) in the C57BL/6 resistant mouse strain. However and in agreement with a number of previous observations indicating that specific CD8 T cells are primed during natural infection or vaccination in humans, a great deal of evidence obtained recently with the susceptible BALB/c murine model of infection by Leishmania major indicates that CD8 T cells participate in both pathogenesis and immunity to cutaneous leishmaniasis. Our goal herein was to identify in silico all parasitic peptides present in the whole L. major predicted proteome, using several public computational systems for the prediction of peptide binding to all MHC (histocompatibility complex-2) molecules in BALB/c and C57BL/6 mice (Syfpeithi, Rankpep, PRED(BALB/c) and Bimas). Peptides that were predicted to bind to different H2 molecules were then analysed for their homology with any of the murine proteins annotated so far, using the BLAST algorithm. Sets of selected peptides for each H2 molecule were defined by different prediction systems and compared to each other. Surprisingly, the results showed that a higher number of L. major peptides were predicted to bind H2 BALB/c molecules and very few or none to bind H2 C57BL/6 molecules. Our finding illustrates how a hybrid immuno computational approach may be useful for biologists to target an in silico set of selected proteins to define potential candidate antigens for experimental vaccination with greater accuracy as well as a reduced number of T cell antigens. PMID- 18420467 TI - Quantum mechanical scattering cross sections and permeability coefficients for ions crossing the human red cell and resting squid axon membranes. AB - Quantum mechanical calculations of elastic scattering cross sections for some permeant ions crossing the human red blood cell and resting axolemma squid axon membranes have been carried out using the three-dimensional spherically symmetric square potential well. Making the assumption that the permeability coefficient is inversely proportional to scattering cross section, we obtain the order of membrane selectivity for the ions as well as values for the permeability coefficients. Despite the relatively simple method used, good agreement between calculated permeability coefficients and data available in the literature is obtained. We suggest that elastic scattering cross section measurements for ions in various membranes would be valuable not only because they give a precise idea about the permeability ratios between ions but they also determine the form of the potential the ions are moving in. PMID- 18420468 TI - Oxidative stress and cystic fibrosis-related diabetes: a pilot study in children. AB - BACKGROUND: Cystic fibrosis (CF) is characterized by chronic inflammation with increased oxidative stress. We evaluated the relationship between glucose tolerance and oxidative stress in CF children. METHODS: Patients 10-18 years old underwent oral glucose tolerance testing (n=31). At 2-h, we assessed blood glutathione and 4-hydroxynonenal-protein adducts (HNE-P), and urine 1,4 dihydroxynonane-mercapturic acid conjugate (DHN-MA). Plasma fatty acid (FA) profile was performed. Patients with impaired glucose tolerance (IGT) were retested 6 to 24 months later and received additional nutritional recommendations (NR) when possible. RESULTS: Fifty-two percent of patients had normal glucose tolerance (NGT), 42% IGT and 6% cystic fibrosis-related diabetes (CFRD). HNE-P concentrations significantly increased with diabetes (109%). Two-h BG correlated positively with HNE-P and negatively with DHN-MA. FA profile was modified with IGT. Of retested IGT patients, 25% received no NR; they remained IGT at 6 months and progressed to CFRD. Of those who received NR, 67% normalized, 11% remained intolerant and 22% developed CFRD. HNE-P levels dropped (88%) in IGT patients reverting to NGT, increased (94%) in the IGT patients with NR developing CFRD, decreased (90%) with persistent IGT. CONCLUSION: CF children showed evidence of increased oxidative stress with worsening of glucose metabolism. NR may delay the appearance of CFRD. PMID- 18420469 TI - Monotone Signal Segments Analysis as a novel method of breath detection and breath-to-breath interval analysis in rat. AB - We applied a novel approach to respiratory waveform analysis--Monotone Signal Segments Analysis (MSSA) on 6-h recordings of respiratory signals in rats. To validate MSSA as a respiratory signal analysis tool we tested it by detecting: breaths and breath-to-breath intervals; respiratory timing and volume modes; and changes in respiratory pattern caused by lesions of monoaminergic systems in rats. MSSA differentiated three respiratory timing (tachypneic, eupneic, bradypneic-apneic), and three volume (artifacts, normovolemic, hypervolemic sighs) modes. Lesion-induced respiratory pattern modulation was visible as shifts in the distributions of monotone signal segment amplitudes, and of breath-to breath intervals. Specifically, noradrenergic lesion induced an increase in mean volume (por=0.06). MSSA of timing modes detected noradrenergic lesion-induced interdependent changes in the balance of eupneic (decrease; p350 mg/dl. Tolerated and rejected grafts were studied histologically and by immunostaining for human T-cells and insulin production. CD4 and CD8 T-cells from the spleen were studied for suppressor activity, expression of cytokines, and CD40L. RESULTS: Although human T-cell engraftment was similar in all groups, ILT3-Fc-treated mice tolerated the islets for the entire period of observation (91 days), whereas control mice rejected the graft within 7 weeks (P < 0.0001). ILT3-Fc treatment suppressed the expression of cytokines and CD40L and induced the differentiation of human CD8(+) T suppressor cells that inhibited Th alloreactivity against graft HLA antigens. T-cells allostimulated in vitro in the presence of ILT3-Fc inhibited CD40L-induced upregulation of CD40 in human pancreatic islet cells. Histochemical studies showed dramatic differences between human pancreatic islets from tolerant, ILT3 Fc-treated mice and control recipients rejecting the grafts. CONCLUSIONS: The data indicated that ILT3-Fc is a potent immunoregulatory agent that suppressed islet allograft rejection in humanized NOD/SCID mice. PMID- 18420486 TI - MAPK kinase kinase-1 is essential for cytokine-induced c-Jun NH2-terminal kinase and nuclear factor-kappaB activation in human pancreatic islet cells. AB - OBJECTIVE: The transcription factor nuclear factor-kappaB (NF-kappaB) and the mitogen-activated protein kinases (MAPKs) c-Jun NH(2)-terminal kinase (JNK) 1/2 are known to play decisive roles in cytokine-induced damage of rodent beta-cells. The upstream events by which these factors are activated in response to cytokines are, however, uncharacterized. The aim of the present investigation was to elucidate a putative role of the MAPK kinase kinase-1 (MEKK-1) in cytokine induced signaling. RESEARCH DESIGN AND METHODS: To establish a functional role of MEKK-1, the effects of transient MEKK-1 overexpression in betaTC-6 cells, achieved by lipofection and cell sorting, and MEKK-1 downregulation in betaTC-6 cells and human islet cells, achieved by diced-small interfering RNA treatment, were studied. RESULTS: We observed that overexpression of wild-type MEKK-1, but not of a kinase dead MEKK-1 mutant, resulted in potentiation of cytokine-induced JNK activation, inhibitor of kappaB (IkappaB) degradation, and cell death. Downregulation of MEKK-1 in human islet cells provoked opposite effects, i.e., attenuation of cytokine-induced JNK and MKK4 activation, IkappaB stability, and a less pronounced NF-kappaB translocation. betaTC-6 cells with a downregulated MEKK 1 expression displayed also a weaker cytokine-induced iNOS expression and lower cell death rates. Also primary mouse islet cells with downregulated MEKK-1 expression were protected against cytokine-induced cell death. CONCLUSIONS: MEKK 1 mediates cytokine-induced JNK- and NF-kappaB activation, and this event is necessary for iNOS expression and cell death. PMID- 18420487 TI - Hyperglycemia-induced reactive oxygen species toxicity to endothelial cells is dependent on paracrine mediators. AB - OBJECTIVE: This study determined the effects of high glucose exposure and cytokine treatment on generation of reactive oxygen species (ROS) and activation of inflammatory and apoptotic pathways in human retinal endothelial cells (HRECs). RESEARCH DESIGN AND METHODS: Glucose consumption of HRECs, human retinal pigment epithelial cells (HRPEs), and human Muller cells (HMCs) under elevated glucose conditions was measured and compared with cytokine treatment. Production of ROS in HRECs was examined using 5-(and-6)-chloromethyl-2',7' dichlorodihydrofluorescein diacetate (CM-H(2)DCFDA), spin-trap electron paramagnetic resonance, and MitoTracker Red staining after high glucose and cytokine treatment. The activation of different signaling cascades, including the mitogen-activated protein kinase pathways, tyrosine phosphorylation pathways, and apoptosis by high glucose and cytokines in HRECs, was determined. RESULTS: HRECs, in contrast to HRPEs and HMCs, did not increase glucose consumption in response to increasing glucose concentrations. Exposure of HRECs to 25 mmol/l glucose did not stimulate endogenous ROS production, activation of nuclear factor-kappaB (NF kappaB), extracellular signal-related kinase (ERK), p38 and Jun NH(2)-terminal kinase (JNK), tyrosine phosphorylation, interleukin (IL)-1beta, or tumor necrosis factor-alpha (TNF-alpha) production and only slightly affected apoptotic cell death pathways compared with normal glucose (5 mmol/l). In marked contrast, exposure of HRECs to proinflammatory cytokines IL-1beta or TNF-alpha increased glucose consumption, mitochondrial superoxide production, ERK and JNK phosphorylation, tyrosine phosphorylation, NF-kappaB activation, and caspase activation. CONCLUSIONS: Our in vitro results indicate that HRECs respond to cytokines rather than high glucose, suggesting that in vivo diabetes-related endothelial injury in the retina may be due to glucose-induced cytokine release by other retinal cells and not a direct effect of high glucose. PMID- 18420488 TI - Human adenovirus type 36 enhances glucose uptake in diabetic and nondiabetic human skeletal muscle cells independent of insulin signaling. AB - OBJECTIVE: Human adenovirus type 36 (Ad-36) increases adiposity but improves insulin sensitivity in experimentally infected animals. We determined the ability of Ad-36 to increase glucose uptake by human primary skeletal muscle (HSKM) cells. RESEARCH DESIGN AND METHODS: The effect of Ad-36 on glucose uptake and cell signaling was determined in HSKM cells obtained from type 2 diabetic and healthy lean subjects. Ad-2, another human adenovirus, was used as a negative control. Gene expression and proteins of GLUT1 and GLUT4 were measured by real time PCR and Western blotting. Role of insulin and Ras signaling pathways was determined in Ad-36-infected HSKM cells. RESULTS: Ad-36 and Ad-2 infections were confirmed by the presence of respective viral mRNA and protein expressions. In a dose-dependent manner, Ad-36 significantly increased glucose uptake in diabetic and nondiabetic HSKM cells. Ad-36 increased gene expression and protein abundance of GLUT1 and GLUT4, GLUT4 translocation to plasma membrane, and phosphatidylinositol 3-kinase (PI 3-kinase) activity in an insulin-independent manner. In fact, Ad-36 decreased insulin receptor substrate-1 (IRS-1) tyrosine phosphorylation and IRS-1-and IRS-2-associated PI 3-kinase activities. On the other hand, Ad-36 increased Ras gene expression and protein abundance, and Ras siRNA abrogated Ad-36-induced PI 3-kinase activation, GLUT4 protein abundance, and glucose uptake. These effects were not observed with Ad-2 infection. CONCLUSIONS: Ad-36 infection increases glucose uptake in HSKM cells via Ras activated PI 3-kinase pathway in an insulin-independent manner. These findings may provide impetus to exploit the role of Ad-36 proteins as novel therapeutic targets for improving glucose handling. PMID- 18420489 TI - Protective role of programmed death 1 ligand 1 (PD-L1)in nonobese diabetic mice: the paradox in transgenic models. AB - OBJECTIVE: Coinhibitory signals mediated via programmed death 1 (PD-1) receptor play a critical role in downregulating immune responses and in maintaining peripheral tolerance. Programmed death 1 ligand 1 (PD-L1), the interacting ligand for PD-1, widely expressed in many cell types, acts as a tissue-specific negative regulator of pathogenic T-cell responses. We investigated the protective potential of PD-L1 on autoimmune diabetes by transgenically overexpressing PD-L1 in pancreatic beta-cells in nonobese diabetic (NOD) mice. RESEARCH DESIGN AND METHODS: We established an insulin promoter-driven murine PD-L1 transgenic NOD mouse model to directly evaluate the protective effect of an organ-specific PD-L1 transgene against autoimmune diabetes. Transgene expression, insulitis, and diabetic incidence were characterized in these transgenic NOD mice. Lymphocyte development, Th1 cells, and regulatory T-cells were analyzed in these transgenic mice; and T-cell proliferation, adoptive transfer, and islet transplantation were performed to evaluate the PD-L1 transgene-mediated immune-protective mechanisms. RESULTS: The severity of insulitis in these transgenic mice is significantly decreased, disease onset is delayed, and the incidence of diabetes is markedly decreased compared with littermate controls. NOD/SCID mice that received lymphocytes from transgenic mice became diabetic at a slower rate than mice receiving control lymphocytes. Moreover, lymphocytes collected from recipients transferred by lymphocytes from transgenic mice revealed less proliferative potential than lymphocytes obtained from control recipients. Transgenic islets transplanted in diabetic recipients survived moderately longer than control islets. CONCLUSIONS: Our results demonstrate the protective potential of transgenic PD-L1 in autoimmune diabetes and illustrate its role in downregulating diabetogenic T-cells in NOD mice. PMID- 18420490 TI - Improved vascular engraftment and graft function after inhibition of the angiostatic factor thrombospondin-1 in mouse pancreatic islets. AB - OBJECTIVE: Insufficient development of a new intra-islet capillary network after transplantation may be one contributing factor to the failure of islet grafts in clinical transplantation. The present study tested the hypothesis that the angiostatic factor thrombospondin-1 (TSP-1), which is normally present in islets, restricts intra-islet vascular expansion posttransplantation. RESEARCH DESIGN AND METHODS: Pancreatic islets of TSP-1-deficient (TSP-1(-/-)) mice or wild-type islets transfected with siRNA for TSP-1 were transplanted beneath the renal capsule of syngeneic or immunocompromised recipient mice. RESULTS: Both genetically TSP-1(-/-) islets and TSP-1 siRNA-transfected islet cells demonstrated an increased vascular density when compared with control islets 1 month after transplantation. This was also reflected in a markedly increased blood perfusion and oxygenation of the grafts. The functional importance of the improved vascular engraftment was analyzed by comparing glucose-stimulated insulin release from islet cells transfected with either TSP-1 siRNA or scramble siRNA before implantation. These experiments showed that the increased revascularization of grafts composed of TSP-1 siRNA-transfected islet cells correlated to increments in both their first and second phase of glucose stimulated insulin secretion. CONCLUSIONS: Our findings demonstrate that inhibition of TSP-1 in islets intended for transplantation may be a feasible strategy to improve islet graft revascularization and function. PMID- 18420491 TI - RAGE and modulation of ischemic injury in the diabetic myocardium. AB - OBJECTIVE: Subjects with diabetes experience an increased risk of myocardial infarction and cardiac failure compared with nondiabetic age-matched individuals. The receptor for advanced glycation end products (RAGE) is upregulated in diabetic tissues. In this study, we tested the hypothesis that RAGE affected ischemia/reperfusion (I/R) injury in the diabetic myocardium. In diabetic rat hearts, expression of RAGE and its ligands was enhanced and localized particularly to both endothelial cells and mononuclear phagocytes. RESEARCH DESIGN AND METHODS: To specifically dissect the impact of RAGE, homozygous RAGE null mice and transgenic (Tg) mice expressing cytoplasmic domain-deleted RAGE (DN RAGE), in which RAGE-dependent signal transduction was deficient in endothelial cells or mononuclear phagocytes, were rendered diabetic with streptozotocin. Isolated perfused hearts were subjected to I/R. RESULTS: Diabetic RAGE-null mice were significantly protected from the adverse impact of I/R injury in the heart, as indicated by decreased release of LDH and lower glycoxidation products carboxymethyl-lysine (CML) and pentosidine, improved functional recovery, and increased ATP. In diabetic Tg mice expressing DN RAGE in endothelial cells or mononuclear phagocytes, markers of ischemic injury and CML were significantly reduced, and levels of ATP were increased in heart tissue compared with littermate diabetic controls. Furthermore, key markers of apoptosis, caspase-3 activity and cytochrome c release, were reduced in the hearts of diabetic RAGE modified mice compared with wild-type diabetic littermates in I/R. CONCLUSIONS: These findings demonstrate novel and key roles for RAGE in I/R injury in the diabetic heart. PMID- 18420492 TI - Dietary phytoestrogens activate AMP-activated protein kinase with improvement in lipid and glucose metabolism. AB - OBJECTIVE: Emerging evidence suggests that dietary phytoestrogens can have beneficial effects on obesity and diabetes, although their mode of action is not known. Here, we investigate the mechanisms mediating the action of dietary phytoestrogens on lipid and glucose metabolism in rodents. RESEARCH DESIGN AND METHODS: Male CD-1 mice were fed from conception to adulthood with either a high soy-containing diet or a soy-free diet. Serum levels of circulating isoflavones, ghrelin, leptin, free fatty acids, triglycerides, and cholesterol were quantified. Tissue samples were analyzed by quantitative RT-PCR and Western blotting to investigate changes of gene expression and phosphorylation state of key metabolic proteins. Glucose and insulin tolerance tests and euglycemic hyperinsulinemic clamp were used to assess changes in insulin sensitivity and glucose uptake. In addition, insulin secretion was determined by in situ pancreas perfusion. RESULTS: In peripheral tissues of soy-fed mice, especially in white adipose tissue, phosphorylation of AMP-activated protein kinase (AMPK) and acetyl CoA carboxylase was increased, and expression of genes implicated in peroxisomal fatty acid oxidation and mitochondrial biogenesis was upregulated. Soy-fed mice also showed reduced serum insulin levels and pancreatic insulin content and improved insulin sensitivity due to increased glucose uptake into skeletal muscle. Thus, mice fed with a soy-rich diet have improved adipose and glucose metabolism. CONCLUSIONS: Dietary soy could prove useful to prevent obesity and associated disorders. Activation of the AMPK pathway by dietary soy is likely involved and may mediate the beneficial effects of dietary soy in peripheral tissues. PMID- 18420493 TI - Pyruvate kinase deficiency and malaria. AB - Malaria that is caused by Plasmodium falciparum is a significant global health problem. Genetic characteristics of the host influence the severity of disease and the ultimate outcome of infection, and there is evidence of coevolution of the plasmodium parasite with its host. In humans, pyruvate kinase deficiency is the second most common erythrocyte enzyme disorder. Here, we show that pyruvate kinase deficiency provides protection against infection and replication of P. falciparum in human erythrocytes, raising the possibility that mutant pyruvate kinase alleles may confer a protective advantage against malaria in human populations in areas where the disease is endemic. PMID- 18420494 TI - A malaria fingerprint in the human genome? PMID- 18420495 TI - Images in clinical medicine. Clonorchis sinensis. PMID- 18420496 TI - Personally controlled online health data--the next big thing in medical care? PMID- 18420497 TI - Off the record--avoiding the pitfalls of going electronic. PMID- 18420498 TI - Physician workforce crisis? Wrong diagnosis, wrong prescription. PMID- 18420499 TI - Weekly paclitaxel in the adjuvant treatment of breast cancer. AB - BACKGROUND: We compared the efficacy of two different taxanes, docetaxel and paclitaxel, given either weekly or every 3 weeks, in the adjuvant treatment of breast cancer. METHODS: We enrolled 4950 women with axillary lymph node-positive or high-risk, lymph node-negative breast cancer. After randomization, all patients first received 4 cycles of intravenous doxorubicin and cyclophosphamide at 3-week intervals and were then assigned to intravenous paclitaxel or docetaxel given at 3-week intervals for 4 cycles or at 1-week intervals for 12 cycles. The primary end point was disease-free survival. RESULTS: As compared with patients receiving standard therapy (paclitaxel every 3 weeks), the odds ratio for disease free survival was 1.27 among those receiving weekly paclitaxel (P=0.006), 1.23 among those receiving docetaxel every 3 weeks (P=0.02), and 1.09 among those receiving weekly docetaxel (P=0.29) (with an odds ratio >1 favoring the groups receiving experimental therapy). As compared with standard therapy, weekly paclitaxel was also associated with improved survival (odds ratio, 1.32; P=0.01). An exploratory analysis of a subgroup of patients whose tumors expressed no human epidermal growth factor receptor type 2 protein found similar improvements in disease-free and overall survival with weekly paclitaxel treatment, regardless of hormone-receptor expression. Grade 2, 3, or 4 neuropathy was more frequent with weekly paclitaxel than with paclitaxel every 3 weeks (27% vs. 20%). CONCLUSIONS: Weekly paclitaxel after standard adjuvant chemotherapy with doxorubicin and cyclophosphamide improves disease-free and overall survival in women with breast cancer. (ClinicalTrials.gov number, NCT00004125 [ClinicalTrials.gov].). PMID- 18420502 TI - Management and outcomes of very low birth weight. PMID- 18420501 TI - Clinical practice. Irritable bowel syndrome. PMID- 18420500 TI - Intensive care for extreme prematurity--moving beyond gestational age. AB - BACKGROUND: Decisions regarding whether to administer intensive care to extremely premature infants are often based on gestational age alone. However, other factors also affect the prognosis for these patients. METHODS: We prospectively studied a cohort of 4446 infants born at 22 to 25 weeks' gestation (determined on the basis of the best obstetrical estimate) in the Neonatal Research Network of the National Institute of Child Health and Human Development to relate risk factors assessable at or before birth to the likelihood of survival, survival without profound neurodevelopmental impairment, and survival without neurodevelopmental impairment at a corrected age of 18 to 22 months. RESULTS: Among study infants, 3702 (83%) received intensive care in the form of mechanical ventilation. Among the 4192 study infants (94%) for whom outcomes were determined at 18 to 22 months, 49% died, 61% died or had profound impairment, and 73% died or had impairment. In multivariable analyses of infants who received intensive care, exposure to antenatal corticosteroids, female sex, singleton birth, and higher birth weight (per each 100-g increment) were each associated with reductions in the risk of death and the risk of death or profound or any neurodevelopmental impairment; these reductions were similar to those associated with a 1-week increase in gestational age. At the same estimated likelihood of a favorable outcome, girls were less likely than boys to receive intensive care. The outcomes for infants who underwent ventilation were better predicted with the use of the above factors than with use of gestational age alone. CONCLUSIONS: The likelihood of a favorable outcome with intensive care can be better estimated by consideration of four factors in addition to gestational age: sex, exposure or nonexposure to antenatal corticosteroids, whether single or multiple birth, and birth weight. (ClinicalTrials.gov numbers, NCT00063063 [ClinicalTrials.gov] and NCT00009633 [ClinicalTrials.gov].). PMID- 18420503 TI - Images in clinical medicine. Lingual thyroid. PMID- 18420504 TI - Case records of the Massachusetts General Hospital. Case 12-2008. A newborn infant with intermittent apnea and seizures. PMID- 18420505 TI - Pharmaceutical promotion to physicians and First Amendment rights. PMID- 18420506 TI - Tectonic shifts in the health information economy. PMID- 18420507 TI - Electronic health records, medical research, and the Tower of Babel. PMID- 18420508 TI - Grassroots activism and the pursuit of an expanded physician supply. PMID- 18420509 TI - Outcome after language mapping for glioma resection. PMID- 18420510 TI - Primary PCI in ST-segment elevation myocardial infarction. PMID- 18420511 TI - Lung transplantation and survival in children with cystic fibrosis. PMID- 18420512 TI - Outcomes of care by hospitalists. PMID- 18420513 TI - Central venous catheterization--subclavian vein. PMID- 18420514 TI - A pivotal medical-device case. PMID- 18420515 TI - Eosinophiluria and acute interstitial nephritis. PMID- 18420516 TI - On continuing education. PMID- 18420517 TI - Noun phrase elaboration in children's spoken stories. AB - PURPOSE: One feature of literate language, noun phrase elaboration, was examined in the oral fictional narratives of school-aged children. METHOD: Two narratives were elicited from 5-, 8- and 11-year-old children, 1 in response to a picture sequence and 1 in response to a single picture. Noun phrases were categorized into 4 types. RESULTS: Simple designating noun phrases were produced by all children at age 5, simple descriptive noun phrases by all children at age 8, and noun phrases with postmodification by all children at age 11. All noun phrase types were produced by more children in object than in subject position and in the single picture than in the picture sequence context. CONCLUSION: There are important developmental changes in noun phrase elaboration in the elementary school years as children learn to manage narrative contexts. Even within picture tasks, variations in visual depiction can affect the use of elaborated noun phrases (ENPs), with more descriptive language more likely to occur in narratives based on a single picture fantasy context than on a realistic picture sequence context. Performance expectations for types of ENPs within these contexts are provided. These findings will be useful to school clinicians in evaluating and working on narrative language within the elementary school period. PMID- 18420518 TI - Prologue: Managing dysphagia in the schools. AB - PURPOSE: This prologue introduces the clinical forum, "Managing Dysphagia in the Schools," and informs the reader of the scope of the included articles. METHOD: The contributing authors are introduced and a brief summary is provided for each of the 10 articles that make up the forum. CONCLUSION: The articles address a broad range of issues, with specific reference to children with dysphagia in school settings. There are comprehensive reviews of legislation, ethical issues, cultural issues, motor learning, medically fragile conditions, severe neuromuscular impairments, and autism spectrum disorder. In addition, there are systematic observations and research that address a model public school dysphagia program, school-based speech-language pathologists' perceptions of competency, family perceptions of school programs, and a program for implementing transition from tube to oral feeding. PMID- 18420519 TI - Legal and financial issues associated with providing services in schools to children with swallowing and feeding disorders. AB - PURPOSE: This review of federal statutes and regulations, state and federal administrative and case law, and policy documents is designed to provide information regarding the treatment of children with swallowing and feeding disorders in the law for the purpose of identifying best practice procedures for school personnel serving children with swallowing and feeding disorders. METHOD: A narrative review of federal statutes and regulations, state and federal administrative and case law, and policy documents was conducted. The review identified prevailing and emerging policies related to serving children with swallowing and feeding issues in schools. In addition, federal standards regarding funding were reviewed for relevance to serving children with dysphagia. IMPLICATIONS: Statutes, regulations, and case law establish the rights of students with health impairments, including swallowing and feeding disorders, to receive such related services as are necessary to attend school safely. Recent administrative hearings have addressed dysphagia, identifying the importance of ensuring the child's feeding safety through diet and services. School districts can receive reimbursement for services that are provided to children with dysphagia through Medicaid when said services are provided in accordance with each state's Medicaid requirements. PMID- 18420520 TI - Ethical issues in providing services in schools to children with swallowing and feeding disorders. AB - PURPOSE: This article is a commentary and discussion of ethical issues in dysphagia services as related to school-based practice in speech-language pathology. METHOD: A review of the literature on ethical issues in the provision of speech-language pathology services to individuals with dysphagia was conducted, with particular emphasis on students receiving school-based services. RESULTS: Issues in dysphagia management that were identified in the literature review are discussed from the perspective of biomedical ethics, professional ethics, and professional practice issues pertinent to the school setting. CONCLUSION: Considerations, suggestions, and resources for ethically responsive action on the part of the school-based speech-language pathologist are provided. PMID- 18420521 TI - Establishing a public school dysphagia program: a model for administration and service provision. AB - PURPOSE: Many school-based speech-language pathologists (SLPs) are hampered in participating in managing children with dysphagia by their school systems' lack of supportive policies and procedures. A need exists to better define the dysphagia-trained SLP's role and clarify the district's responsibility. The purpose of this article is to address the critical administrative issues and administrative components surrounding dysphagia in the schools and to offer the SLP some solutions to these problems. METHOD: A narrative review of the relevant literature addresses the following questions that are important for administrative planning and implementation of programs for students with dysphagia in the schools: (a) Should school systems assume responsibility for working with students with dysphagia? (b) Why is a system-supported procedure recommended? (c) What service delivery models can a system use to serve students with dysphagia? (d) What components should be included in a dysphagia procedure? CONCLUSION: SLPs should be proactive in providing dysphagia services to students in the schools. A procedure that is adopted by a school system for all of its employees to follow will offer direction and guidance. This system-supported procedure may provide assurance that dysphagia services are being provided in a professionally acceptable manner. Ongoing staff development and training is essential. A system can implement dysphagia services using existing staff, when possible, and policies and procedures that have been approved by the system. PMID- 18420522 TI - Training and self-reported confidence for dysphagia management among speech language pathologists in the schools. AB - PURPOSE: The number of children requiring dysphagia management in the schools is increasing. This article reports survey findings relative to speech-language pathologists' (SLPs') training and self-rated confidence to treat children with swallowing and feeding disorders in the schools. METHOD: Surveys were completed by 222 SLPs representing Virginia and its contiguous states. Queries on dysphagia training targeted formal education, on-the-job experiences, and current caseload information. In addition, participants self-rated their confidence to treat dysphagia. RESULTS: Statistically significant relationships between training and self-confidence levels were demonstrated. Specifically, participation in continuing education and currency of educational activities revealed significant and moderately strong correlations to self-reported confidence to treat children with dysphagia in the school setting. CONCLUSION: Findings support continuing education as a correlate to self-reported confidence to treat dysphagia in the school setting among SLPs in Virginia and its contiguous states. Further research is merited to ascertain if these findings reflect national trends. Quantifiable, cost-effective, and evidenced-based dysphagia training, consultancy, and management models are needed if school-based SLPs are to meet the increasing challenges of their diverse caseloads. PMID- 18420523 TI - Family and cultural issues in a school swallowing and feeding program. AB - PURPOSE: This article presents a rationale for speech-language pathologists (SLPs) to provide culturally competent evaluation, diagnostic, and intervention services for children with oral motor, swallowing, and feeding disorders in school settings. There is also a discussion of how changing American public school demographics necessitate the consideration of cultural issues and family focused approaches to dysphagia services. METHOD: This article provides an overview of cultural, religious, and health beliefs of several ethnic groups, as well as information on the dynamics and requirements of cultural competence and family-focused intervention. Ethnographic interviewing is presented as a culturally sensitive diagnostic method. Information on structuring a culturally relevant individualized educational program process is provided. Guidance in the use of interpreters and translators is also offered. CONCLUSION: Attention to cultural and family issues in the diagnosis and treatment of dysphagia in a school setting optimizes the opportunities for successful outcomes and better meets the needs of children and families from culturally/linguistically diverse backgrounds. PMID- 18420524 TI - Family perceptions of facilitators and inhibitors of effective school-based dysphagia management. AB - PURPOSE: This qualitative study focused on the perceptions of family members of children with dysphagia by asking what the family-identified factors are that facilitate or inhibit effective school-based management of pediatric dysphagia. METHOD: Semistructured interviews of 7 family members of 6 children with dysphagia, ages 2 through 11 years, were conducted. Collective case study and cross-case analyses were conducted. The findings were confirmed through triangulation, respondent validation, and member checking. RESULTS: Interviewees identified 5 facilitating factors that enhance school-based dysphagia programs: (a) setting, (b) therapist and/or program, (c) capitalizing on positive child characteristics, (d) home-school interactions, and (e) external supports. Four factors that inhibit effectiveness also emerged: (a) setting, (b) therapist and/or program, (c) limiting child characteristics, and (d) home-school interactions. CONCLUSION: Practitioners may consider addressing the factors that were identified as facilitators of effective programs. More research is needed concerning (a) factors promoting effective parent-professional relationships, (b) methods for increasing professional preparation in pediatric dysphagia, and (c) the efficacy of school-based pediatric dysphagia management programs. PMID- 18420525 TI - Using motor learning approaches for treating swallowing and feeding disorders: a review. AB - PURPOSE: This article discusses children's development of mature swallowing and feeding as a process of skill acquisition and considers the applicability of motor learning concepts for advancing these capabilities in school-aged children. METHOD: The motor learning literature was reviewed, with concentration on (a) concepts that are relevant for the acquisition of skill and (b) structuring practice experiences to optimize learning. RESULTS: The discussion includes (a) swallowing physiology with a focus on motor task components, (b) normal development of eating skills, (c) factors that may slow or disrupt normal development, (d) motor learning concepts found to influence learning efficiency and performance adequacy, (e) applications to the assessment and treatment of pediatric swallowing and feeding disorders, and (f) an illustrative case history. CONCLUSION: Deficiencies in swallowing and feeding may encompass eating, saliva control, swallowing during oral hygiene, and swallowing medications. Motor learning literature provides a rich foundation of evidence-based theory and educational strategies for the development and remediation of motor-based skills such as swallowing and feeding. PMID- 18420526 TI - Schoolchildren with dysphagia associated with medically complex conditions. AB - PURPOSE: This article reviews population trends and general characteristics of children with dysphagia in schools, provides an overview of dysphagia teams and the roles of the speech-language pathologist (SLP) in school and hospital settings, and describes assessment and treatment of swallowing and feeding problems in children with complex medical histories. METHOD: A review of the literature of swallowing and feeding problems in young children and population trends for children who are at risk for dysphagia was completed. Two case studies are presented to illustrate the complexities that are common to this population and to provide practical information to maximize the health and education outcomes of children with dysphagia. RESULTS: SLPs in schools are increasingly evaluating and treating children with dysphagia associated with medically complex conditions. There is a rising incidence in preterm births and the survival of medically fragile children, suggesting that a greater number of children will be at risk for dysphagia in the future. Limited evidence is available to support most therapeutic interventions. CONCLUSION: School-based SLPs are uniquely positioned to identify swallowing and feeding problems, evaluate and treat children with dysphagia, participate on dysphagia teams in the school setting, and interact with the medical team. PMID- 18420527 TI - Transition from tube to oral feeding in the school setting. AB - PURPOSE: A school-based treatment program for tube-fed children with medically complex conditions and food refusal was implemented to facilitate the children's transition to oral feeding and advance their eating skills. METHOD: The program combined educational and therapeutic goals. It was implemented in a regional public school for children with hearing impairments. A team approach was used. Collaboration with the student's families, medical care providers, personal assistants, and classroom staff was maintained. RESULTS: Detailed case reviews are provided for 2 of the children who completed the program at age 8;1 (years;months) and 7;8 after 29 and 26 months of treatment, respectively. At the end of the program, tube feeding was discontinued; the children were feeding themselves and taking medications orally. One child was using mature eating skills to eat an unrestricted diet; the other was eating a modified diet consisting of a full nutrient drink and limited smooth and lightly textured solids. CONCLUSION: Successful transition to oral feeding and advancement of developmental eating skills were accomplished in a school setting for long-term tube-fed children with medically complex conditions. The program included collaboration with family and medical personnel as well as integrated therapeutic and educational goals. PMID- 18420528 TI - Addressing feeding disorders in children on the autism spectrum in school-based settings: physiological and behavioral issues. AB - PURPOSE: The purposes of this article are to define the nature of feeding difficulties in children with autism spectrum disorder (ASD), identify important components of the assessment and treatment of feeding disorders specific to this population, and delineate specific therapeutic techniques designed to improve assessment and treatment within the school setting. METHOD: Literature review and case example are used to define the predominant nature of the feeding difficulties that are experienced by some children on the autism spectrum. Characteristics of this complex disorder that can have an impact on feeding skill and behavior are also identified. These factors are then integrated to create assessment and intervention techniques that can be used in conjunction with traditional feeding approaches to facilitate improvements in eating in this unique population. IMPLICATIONS: The complex nature of ASD and its many influences on feeding skills and behavior create the need for modification to both assessment and treatment approaches. Additional research is needed to create therapeutic protocols that can be used by school-based speech-language pathologists to effectively assess and treat feeding difficulties that are commonly encountered in children with ASD. PMID- 18420529 TI - Epilogue: Managing dysphagia in the schools. AB - PURPOSE: This epilogue summarizes the contribution of the articles in the clinical forum, "Managing Dysphagia in the Schools" and discusses future trends in pediatric dysphagia and the need for clinical practice and research. METHOD: The contribution of each of the 10 articles that make up the forum is summarized briefly. CONCLUSION: This clinical forum provides a discussion of numerous issues that need to be considered by clinicians working with children who exhibit swallowing and feeding problems and their families in school settings. Current and future challenges exist for clinicians, children, and families dealing with dysphagia. Discussion of current issues and strategies for successful treatment outcomes are provided. PMID- 18420530 TI - Hyperventilation and cerebrospinal fluid acidosis caused by topiramate. AB - OBJECTIVE: To report a case of hyperventilation caused by topiramate therapy and propose a pathophysiologic mechanism for this disorder. CASE SUMMARY: A 52-year old woman with refractory seizure disorder was admitted to the burn care unit with burns over 10% of her body. Her seizure medications, unchanged and well tolerated for several months, included carbamazepine 1200 mg, lamotrigine 500 mg, phenobarbital 80 mg, and topiramate 150 mg per day. During hospitalization, despite a relatively normal arterial pH, the woman developed persistent hyperventilation, with respiratory rates up to 50 breaths/min. Alkalinization did not reduce the hyperventilation. Thoracic contrast-enhanced computed tomographic scan ruled out pulmonary embolism and persistent pneumonia. Salicylate and biguanide screening were negative; results of repeated thyroid and liver function tests were normal. Cerebral magnetic resonance imaging excluded a cerebral pathology. After cerebrospinal fluid (CSF) analysis showed acidosis (pH 7.14), topiramate was withdrawn and the patient's general condition rapidly improved. Forty-eight hours later, the CSF pH had increased to 7.26. The woman was discharged from the burn care unit on the 42nd hospital day. DISCUSSION: Hyperchloremic normal anion gap metabolic acidosis, which can lead to hyperventilation, has been reported as an adverse effect of topiramate treatment. However, our patient had respiratory alkalosis. Concurrent etiologies of peripheral hyperventilation were excluded, leaving central neurogenic hyperventilation as the remaining etiology. Such central neurogenic hyperventilation associated with topiramate has previously been reported in intensive care. Our case report demonstrates CSF acidosis. Withdrawing topiramate reduced both CSF acidosis and hyperventilation. The mechanism of topiramate induced CSF acidosis remains unclear. According to the Naranjo probability scale, the relationship of hyperventilation to administration of topiramate in our patient was probable. CONCLUSIONS: Normal doses of topiramate may provoke central neurogenic hyperventilation, as a result of CSF acidosis. The acid-base status of critically ill patients receiving topiramate should be monitored carefully. PMID- 18420531 TI - Population pharmacokinetic and pharmacodynamic analysis of pegloticase in subjects with hyperuricemia and treatment-failure gout. AB - Pegloticase is designed to convert urate into the easily excretable allantoin to treat hyperuricemia in gout. The aim of this analysis was to describe the pharmacokinetics and pharmacodynamics of pegloticase in 40 gout patients. Pegloticase was administered as intravenous infusions every 2 weeks at 4- and 8 mg doses or every 4 weeks at 8- or 12-mg doses for 12 weeks. Serum pegloticase concentrations, plasma urate, and serum antibody response were determined. Population pharmacokinetics and pharmacodynamics analyses were performed. Data were modeled simultaneously, and covariates were investigated (age, gender, race, body weight, ideal body weight, and antibody response). The dosing regimens to maintain uric acid levels below the therapeutic target of 6 mg/dL were then predicted by the model. The pharmacokinetics were best described by a 1 compartment linear model, while the pharmacodynamics model was fitted as a direct effect of pegloticase on uric acid concentrations with a suppressive maximum effect attributed to drug (E(max)) function. Pegloticase suppressed uric acid levels up to 83%. Weight only affected clearance and volume of distribution. No covariates affected pharmacodynamics. Simulation suggests pegloticase administered at 8 mg every 2 or 4 weeks as 2-hour intravenous infusions will maintain uric acid levels well under 6 mg/dL. PMID- 18420532 TI - Limitations of using a single postdose midazolam concentration to predict CYP3A mediated drug interactions. AB - Midazolam is a common probe used to predict CYP3A activity, but multiple blood samples are necessary to determine midazolam's area under the concentration-time curve (AUC). As such, single sampling strategies have been examined. The purpose of this study was to assess the ability of single midazolam concentrations to predict midazolam AUC in the presence and absence of CYP3A modulation by Ginkgo biloba extract (GBE). Subjects received oral midazolam 8 mg before and after 28 days of GBE administration. Postdose blood samples were collected during both study periods and midazolam AUC determined. Linear regression was used to generate measures of predictive performance for each midazolam concentration. The geometric mean ratio (90% confidence intervals) of midazolam AUC(0-infinity) post GBE/AUC(0-infinity) pre-GBE was 0.66 (0.49-0.84) (P = .03). Before and after GBE administration, optimal midazolam sampling times were identified at 3.5 to 5 hours and 2 to 3 hours, respectively. Single midazolam concentrations between 2 and 5 hours correctly predicted the reduction in midazolam AUC following GBE exposure, but confidence intervals were generally wide. Intersubject variability in CYP3A activity (either inherent or from drug administration) alters the prediction of optimal midazolam sampling times; therefore, midazolam AUC is preferred for assessing CYP3A activity in drug-drug interaction studies. PMID- 18420534 TI - Excluding qualitative inquiry: an open letter to the Canadian Urological Association. PMID- 18420533 TI - The effect of IL6-174C/G polymorphism on postprandial triglyceride metabolism in the GOLDN studyboxs. AB - Chronically elevated interleukin-6 (IL-6) affects lipid and lipoprotein metabolism. Individuals genetically predisposed to higher IL-6 secretion may be at risk of dyslipidemia, especially during the postprandial phase. We investigated the effect of genetic variants at the IL6 locus on postprandial lipemia in US Whites participating in the Genetics of Lipid Lowering Drugs and Diet Network study. Subjects were given a single fat load composed of 3% of calories as protein, 14% as carbohydrate, and 83% as fat. Blood was drawn at 0 h, 3.5 h, and 6 h to determine plasma triglyceride (TG), TG-rich lipoprotein (TRL) and lipoprotein particle size. Homozygotes (GG) and heterozygotes (CG) of the 174C/G variant displayed higher plasma IL-6 concentrations compared with major allele homozygotes (CC) (P = 0.029). GG and CG subjects showed higher fasting plasma TG (P = 0.025), VLDL (P = 0.04), and large VLDL (P = 0.02) concentrations than did CC subjects. Moreover, GG and CG subjects experienced greater postprandial response of TG (P = 0.006) and TRL, including chylomicrons (P = 0.005), total VLDL (P = 0.029), and large VLDL (P = 0.017) than did CC subjects. These results suggest that the functional polymorphism -174C>G at the IL6 locus determines the difference in both fasting and postprandial TG metabolism. This phenomenon could be responsible for the observed association of this genetic variant with cardiovascular disease risk. PMID- 18420535 TI - To discipline a "dog": the essential structure of mastering diabetes. AB - In this article we describe the findings of a study in which the essential structure of mastering diabetes was explored from the patient's viewpoint, emphasizing the lived experience of adhering and not adhering to a complex treatment regimen for diabetes. PMID- 18420536 TI - Long-term sickness absence due to burnout: absentees' experiences. AB - In the late 1990s, there was a marked increase in Sweden in long-term sickness absence with mental diagnoses. In the extensive research on burnout, little attention has been paid to the link with sickness absence. Our aim in this study was to discover what caused burnout, or such severe stress that it led to long term sickness absence (> 28 days). We interviewed 32 individuals on long-term sickness absence with a diagnosis of burnout and used grounded theory to analyze the data. We believe that events prior to the sickness absence can be understood as a process of emotional deprivation, and we described the process in terms of a flight of stairs with eight steps: "The Burnout Stairs." We argue that the emotion conveying the process of emotional deprivation is unacknowledged shame, which has its basis in poor social relations at work. We discuss the study's limitations and the need for further research in this field. PMID- 18420537 TI - Listening to rural Hispanic immigrants in the Midwest: a community-based participatory assessment of major barriers to health care access and use. AB - Hispanic immigrants are increasingly residing in rural communities, including in the midwestern United States. Limitations in the ability of rural Hispanics to access and utilize health care contribute to patterns of poor health and health disparity. A conceptual model of "vulnerability" guides this community-based participatory assessment project designed to explore rural Hispanics' perceived barriers to accessing and utilizing health care. Findings from a series of 19 focus groups with 181 participants from three communities in the upper Midwest identified perceived barriers at the individual and health care system levels. The most commonly perceived barriers were the lack of and limitations in health insurance coverage, high costs of health care services, communication issues involving patients and providers, legal status/discrimination, and transportation concerns. Findings imply that these barriers could be addressed using multiple educational and health service delivery policy-related strategies that consider the vulnerable nature of this growing population. PMID- 18420538 TI - I have mastered the challenge of living with a chronic disease: life stories of people with rheumatoid arthritis. AB - Our aim with this study was to explore the narrative life story of individuals diagnosed with rheumatoid arthritis. An open qualitative approach, namely narrative biographic methodology, was applied to include the life context of the participants and to deliberately exclude predefinitions of concepts. Ten people with rheumatoid arthritis who retired early because of the disease participated and were interviewed three times according to a narrative biographic interview style. The biographical data and the interview texts were analyzed both individually and in comparison to each other. Some participants regarded rheumatoid arthritis as a challenge for mastery in their lives, whereas others adapted to the disease and "made the best out of a bad situation." Especially in countries where the medical model predominates in health care, our findings can be used to broaden the current view that some health professionals have toward patients, and stress the importance of patients being self-responsible. PMID- 18420539 TI - Developing theory from complexity: reflections on a collaborative mixed method participatory action research study. AB - Research studies are increasingly complex: They draw on multiple methods to gather data, generate both qualitative and quantitative data, and frequently represent the perspectives of more than one stakeholder. The teams that generate them are increasingly multidisciplinary. A commitment to engaging community members in the research process often adds a further layer of complexity. How to approach a synthesizing analysis of these multiple and varied data sources with a large research team requires considerable reflection and dialogue. In this article, we outline the strategies used by one multidisciplinary team committed to a participatory action research (PAR) approach and engaged in a mixed method program of research to synthesize the findings from four subprojects into a conceptual framework that could guide practice in community mental health organizations. We also summarize factors that hold promise for increasing productivity when managing complex research projects. PMID- 18420540 TI - In search of respect for qualitative research. PMID- 18420541 TI - Making behavioral activation more behavioral. AB - Behavioral Activation, an efficacious treatment for depression, presents a behavioral theory of depression--emphasizing the need for clients to contact positive reinforcement--and a set of therapeutic techniques--emphasizing provision of instructions rather than therapeutic provision of reinforcement. An integration of Behavioral Activation with another behavioral treatment, Functional Analytic Psychotherapy, addresses this mismatch. Functional Analytic Psychotherapy provides a process for the therapeutic provision of immediate and natural reinforcement. This article presents this integration and offers theoretical and practical therapist guidelines on its application. Although the integration is largely theoretical, empirical data are presented in its support when available. The article ends with a discussion of future research directions. PMID- 18420542 TI - Positive behavior support and applied behavior analysis: a familial alliance. AB - Positive behavior support (PBS) emerged in the mid-1980s as an approach for understanding and addressing problem behaviors. PBS was derived primarily from applied behavior analysis (ABA). Over time, however, PBS research and practice has incorporated evaluative methods, assessment and intervention procedures, and conceptual perspectives associated with a number of additional disciplines. Recently, there has been some confusion regarding the definition of PBS and, in particular, its relationship to ABA. In this article, it was noted that the practice of PBS and ABA, in some instances, can be indistinguishable but that important differences in definitions and emphases mandate an explicit distinction. The purpose of this article is to address some of the key points of confusion, identify areas of overlap and distinction, and facilitate a constructive and collegial dialog between proponents of the PBS and ABA perspectives. PMID- 18420543 TI - Efficacy of an emotion-focused treatment for prolonged fatigue. AB - Previous research findings have suggested a relationship between less adaptive emotional functioning and fatigue. The present study used a research design involving multiple baselines across participants to evaluate the efficacy of a new emotion-focused treatment for prolonged fatigue delivered in a cognitive behavioral therapy framework. The 13 adults participating in the study met the criteria for prolonged fatigue and provided fatigue baselines of 2, 5, or 8 weeks. The results indicated that the treatment was effective, with fatigue severity levels after the initiation of treatment significantly lower than that predicted by baseline patterns, as determined by the split median method of trend estimation. At 3-4 months after treatment, 8 of 11 clients who completed the treatment no longer met the criteria for prolonged fatigue. PMID- 18420544 TI - Alcohol drinking and colorectal cancer in Japanese: a pooled analysis of results from five cohort studies. AB - Colorectal cancer is an alcohol-related malignancy; however, the association appears to be stronger among Asian populations with a relatively high prevalence of the slow-metabolizing aldehyde dehydrogenase variant. To examine the association between alcohol consumption and colorectal cancer in Japanese, the authors analyzed original data from five cohort studies that measured alcohol intake using validated questionnaires at baseline. Hazard ratios were calculated in the individual studies, with adjustment for a common set of variables, and then combined using a random-effects model. During 2,231,010 person-years of follow-up (ranging variously from 1988 to 2004), 2,802 colorectal cancer cases were identified. In men, multivariate-adjusted pooled hazard ratios for alcohol intakes of 23-45.9 g/day, 46-68.9 g/day, 69-91.9 g/day, and > or =92 g/day, compared with nondrinking, were 1.42 (95% confidence interval (CI): 1.21, 1.66), 1.95 (95% CI: 1.53, 2.49), 2.15 (95% CI: 1.74, 2.64), and 2.96 (95% CI: 2.27, 3.86), respectively (p for trend < 0.001). The association was evident for both the colon and the rectum. A significant positive association was also observed in women. One fourth of colorectal cancer cases in men were attributable to an alcohol intake of > or =23 g/day. An alcohol-colorectal cancer association seems to be more apparent in Japanese than in Western populations. Whether this difference can be ascribed to genetic or environmental factors needs to be clarified. PMID- 18420547 TI - Rethinking the conceptual and empirical foundations of clinical ethics. AB - The five papers in the 2008 "Clinical Ethics" number of the journal address the conceptual and empirical foundations of clinical ethics. Three articles take up the concept of professionalism in medicine, exploring its possibilities and implications. The fourth article provides a distinctive, phenomenological account of the "placebo effect," a vexing topic of surprising durability in the clinical setting. The final article, a systematic review of the qualitative literature on bedside rationing of resources, creates an empirical foundation for philosophical analysis and argument of a distinctive kind. PMID- 18420548 TI - Professionalism's facets: ambiguity, ambivalence, and nostalgia. AB - Medical educators invoke professionalism as a core competency in curricula. This paper criticizes classic definitions. It also identifies some negative traits of medicine as a profession. The call to professionalism is naive nostalgia. Straightforward didactics in professionalism cannot do the desired work in medical education. The most we can say is that students should adopt the good aspects of professionalism and the profession should stop being some of what it has been. This is a platitude. If the notion is to be more than shallow, each student and practitioner will have to engage in much dialogue, reflection and refinement over many years. PMID- 18420549 TI - Moral damage to health care professionals and trainees: legalism and other consequences for patients and colleagues. AB - Health care professionals' and trainees' conceptions of their responsibilities to patients can change over time for a number of reasons: evolving career goals, desires to serve different patient populations, and changing family obligations, for example. Some changes in conceptions of responsibility are healthy, but others express moral damage. Clinicians' changes in their conceptions of what they are responsible for express moral damage when their responses to others express a meager, rather than robust, sense of what they owe others. At least two important expressions of moral damage in the context of health care are these: callousness and divestiture. Callousness describes the poor condition of a clinician's capacity for moral perception; when her capacity to accurately appreciate features of moral relevance that configure others' needs, vulnerabilities, and desert of care diminishes, such that she fails to respond with care to those for whom she has duties to care, she is callous. Callousness has been explored in detail elsewhere,1 and so the focus of this paper is divestiture. A clinician divests when the value of responding with care to others becomes less centrally and importantly constitutive of his personal and professional identity. Divestiture has important consequences for patients and health professions education, which I will explore here. PMID- 18420550 TI - An analysis of candidate ethical justifications for allowing inexperienced physicians-in-training to perform invasive procedures. AB - Allowing relatively inexperienced physicians-in-training to perform invasive medical procedures is a widely accepted practice, generally felt to be justified by the need to train future generations of physicians. The ethical justification of this practice, however, is rarely if ever explored in any depth. This essay examines the moral issues associated with this practice, in the setting of a specific clinical scenario involving the emergency intubation of a critically ill newborn. The practice is ultimately shown to be justified based not only on the needs of society and future patients but also on the best interests of the patient being treated. However, several important qualifications need to be satisfied in order for this practice to be ethically permissible. The arguments and qualifications presented can be extended to clinical situations beyond the specific scenario discussed and are relevant to a wide range of medical and surgical settings. PMID- 18420551 TI - A phenomenology of the 'placebo effect': taking meaning from the mind to the body. AB - Most mainstream attempts to understand the "placebo effect" invoke expectancy theory, arguing that expecting certain outcomes from a treatment or intervention can manifest those outcomes. Expectancy theory is incompatible with the phenomena of placebo responses, more appropriately named "meaning responses." The expectancy account utilizes reflexive consciousness to connect a world of conceptual representations to mechanical physiology. An alternative account based upon Merleau-Ponty's motor intentionality argues that the body understands and is capable of responding to meanings without the need for any conceptual or linguistic content. A motor intentional framework of meaning poses dramatic implications for the interpretation of clinical trials and in the clinical practice of medicine. Most strongly, it argues that the empathic physician can facilitate the physiologic effects of treatments through skillful participation and manipulation of the meaning response. PMID- 18420552 TI - How physicians allocate scarce resources at the bedside: a systematic review of qualitative studies. AB - Although rationing of scarce health-care resources is inevitable in clinical practice, there is still limited and scattered information about how physicians perceive and execute this bedside rationing (BSR) and how it can be performed in an ethically fair way. This review gives a systematic overview on physicians' perspectives on influences, strategies, and consequences of health-care rationing. Relevant references as identified by systematically screening major electronic databases and manuscript references were synthesized by thematic analysis. Retrieved studies focused on themes that fell under three major headings: (i) conditions and influences of BSR, (ii) strategies of BSR, and (iii) consequences of BSR. The range of themes indicates that physicians' rationing behavior is highly variable, strongly influenced by context-related factors, and consists mainly of implicit rationing strategies. Torn between patient advocacy and the obligation to contain costs, physicians experience various role conflicts. The development of explicit rationing strategies seems necessary to avoid arbitrary BSR and allow a fair allocation of health-care resources. PMID- 18420554 TI - "I thought . . . maybe this is my chance": sexual abuse against girls and women with disabilities in Malawi. AB - This article is based on a pilot study conducted in Malawi in 2006 that intended to uncover episodes of violence and abuse against women and girls with disabilities and furthermore to explore the mechanisms behind such acts. The stories of 23 women with disabilities were obtained through in-depth semistructured interviews that covered, among other topics, aspects of sexual abuse experienced by or known to these women. None of the informants reported having been sexually abused during childhood. In adulthood, several had experienced what they themselves defined as sexual abuse: Men came and wanted to "marry" them. When the woman became pregnant, the man disappeared and left her alone. The interviewees were very opinionated regarding this theme and sought both social and political action in this matter, especially a means to enforce men to take economic responsibility for their biological children. The informants stressed that adapted education for women with disabilities would allow them to become more economically independent and be better able to refuse the advances of unserious suitors. PMID- 18420555 TI - The Rockwood Preparatory Program for sexual offenders: description and preliminary appraisal. AB - This article describes the Rockwood Preparatory Program for sexual offenders. This program operates in the Millhaven induction center of the Correctional Service of Canada. Clients remain in the program for 6 to 8 weeks, depending on when they are placed in their home prison, where they will receive a full treatment program. The preparatory program takes a motivational approach, integrating several theoretical views and employing the therapeutic approaches that have been shown to maximize effectiveness with sexual offenders. The content and approach of the program are described and the observed benefits are outlined. PMID- 18420556 TI - The relationship between victim age and gender crossover among sex offenders. AB - Some child molesters abuse children of both genders. The purpose of this study was to explore the relationship between gender crossover and victim age. The authors hypothesized that offenders who molested children of both genders would be more likely to have very young victims and that offenders who molested very young children would be more likely to abuse children of both sexes than offenders with older victims. The sample consisted of 362 sex offenders being considered for civil commitment as sexually violent predators in Florida. Data were collected from file reviews. The relationship between victim age and gender crossover was examined using chi-square and multivariate logistic regression analyses. The proportion of offenders with victims of both genders significantly increased as the victims' ages decreased, and sex offenders with preschool-age victims were most likely to have abused both boys and girls. A sex offender with a victim 6 years of age or younger had more than 3 times the odds of having perpetrated sex crimes against both genders than a sex offender with only older victims. Sex offenders with victims of both genders had more than 3 times the odds of having preschool victims. The presence of a major mental illness increased the odds of gender crossover more than threefold. Pedophilia increased the odds of having victims younger than 6 more than 11-fold. Implications for practice are discussed. PMID- 18420557 TI - Social desirability and sexual offenders: a review. AB - Social desirability--the desire to make a favorable impression on others-poses a significant threat to the validity of self-reports. This review examines research on social desirability in both forensic and nonforensic populations with the goal of identifying how best to minimize threats to the validity of research with sexual offenders. Although social desirability has long been a major research topic in personality, consensus has not been reached on key questions such as its dimensional structure and whether social desirability constitutes a trait or a response bias. Research with offenders has shown that social desirability is negatively related to recidivism and that different offender subtypes vary in the degree to which social desirability appears to influence self-reports, with child molesters exhibiting the strongest tendency to "fake good." Several methods of controlling for social desirability have been proposed, but the effectiveness of these methods in increasing validity of offender self reports is questionable. Given the lack of consensus in the personality literature, a fresh start is needed in which basic questions regarding social desirability are revisited with respect to offender populations. PMID- 18420558 TI - Improving the measurement of criminal sexual behavior: the application of randomized responding technique. AB - Randomized responding technique (RRT), a method for maintaining anonymity, was used with a sample of sexual offenders recruited into a treatment outcome study. Prior to release from incarceration, three groups--those randomly assigned to treatment, those randomly assigned to no-treatment control, and those who refused participation in the treatment study but consented to a prerelease interview- participated in an alternate-questions RRT procedure. This procedure pairs a nonsensitive question with the sensitive question of interest, in this case, the number of prior sex offenses. Respondents answered either the sensitive or nonsensitive question, depending on the results of a randomizing device (roll of dice). The distributions for the nonsensitive questions were highly skewed; therefore, the outliers were removed and RRT estimates calculated. RRT estimates of prior offending (2.20 prior offenses) were significantly higher than officially recorded prior offenses (0.51 prior offenses). The pattern of differences between treatment and control groups were similar in RRT estimates and officially recorded priors, as were the correlations between RRT estimates and other self-report scales. These results provide preliminary evidence that RRT is a useful method for generating reoffending data that are more sensitive than officially recorded offenses and that contain less bias than other self-reports. PMID- 18420559 TI - An exploratory evaluation of the contribution of personality and childhood sexual victimization to the development of sexually abusive behavior. AB - Scores on the Millon Adolescent Clinical Inventory (MACI) and the Childhood Trauma Questionnaire (CTQ) were collected from two samples of incarcerated delinquent males: 74 adjudicated sexual abusers and 53 nonsexual abusers. In comparison to non-sexually abusive youths, a larger percentage of the sexually abusive youths indicated a history of childhood sexual victimization. The scores were then entered into a logistic regression to assess the contribution of personality characteristics to a social learning "victim-to-victimizer" etiological model. After controlling for time in treatment, the model correctly classified 76.6% of the youths. The variables that contributed significantly to the social learning model were the CTQ Sexual Abuse and Physical Neglect scales and the Submissive and Forceful scales on the MACI. Research and treatment implications of these findings are discussed. PMID- 18420560 TI - Investigation of mean glandular dose versus compressed breast thickness relationship for mammography. AB - The relationship between the mean glandular dose (MGD) and the compressed breast thickness (CBT) is commonly used for the presentation of mammographic dose survey results and could also be useful for the assessment of individual breast doses retrospectively in case of lack of necessary dosimetric instrumentation. The high data scattering from the best fit reduces the reliability of this technique. The aim of this study was to investigate the accuracy of this relationship using the data collected from a patient survey and phantom experiment. Patients were divided into three different groups according to their breast glandularities, which were predicted from the inspection of previous mammograms. X-ray beam qualities that will be used in patient examinations were determined according to breast thickness and predicted glandularities. The MGD versus CBT relationship for all the examined patients resulted in a poor correlation (R2 = 0.28). This relationship was separately obtained for each glandularity group and also for sub groups of specific beam qualities. The best correlation (R(2) = 0.73) was obtained for the fatty breast group and Mo/Mo combination. A low correlation (R2 = 0.34) was observed in the mid-glandularity group due to inclusion of a wide range of glandularities in this group. In the case of the dense breast group, although the glandularity range was narrow, there were e still high data scattering (R2 = 0.25). This was probably due to the use of Mo/Rh and Mo/Mo combinations. This is validated by obtaining the MGD-CBT relationship specific to Mo/Mo combination (R2 = 0.61). PMID- 18420561 TI - Dose rate dependency of electronic personal dosemeters measuring X- and gamma-ray radiation. AB - Three models of electronic personal dosemeters (EPDs)-Siemens Mk 2.3, Rados RAD 60S and Vertec Bleeper Sv-were irradiated with seven photon beam qualities: 60Co, 137Cs and the ISO narrow spectrum series X-ray qualities N-250, N-200, N-150, N 60 and N-20. The personal dose equivalent rates delivered to the devices varied between 0.002 and 0.25 mSv s(-1). Measurements were made with the EPDs mounted free-in-air as well as against Lucite and water phantoms. Results for all models of EPDs showed differences in personal dose equivalent energy response for different energies covered by this range of radiation qualities, with different models showing variations from 15 to 65%. In some cases, the personal dose equivalent rate response of these devices varied by a factor of 3 between irradiations at typical calibration dose rates and those normally encountered by nuclear energy workers. PMID- 18420562 TI - Radon carcinogenesis: risk data and cellular hits. AB - Abundant epidemiological data are now available (2008) on the human lung cancer response for lifetime radon gas exposure to residential concentrations of 100 Bq m(-3), equal to 22 working level months over 40 y. We combined published pooled epidemiological data and dosimetric calculations of alpha particle hits to target basal or mucous cell nuclei in bronchial epithelium. This yields an estimate that about 10,000 basal nuclei (target) cell hits per cm2 per person over a lifetime are involved in radon-related lung cancer. The DNA target cell area (cross section) for a hit is about 2 bp. The present epidemiology indicates that 1000 persons need to be exposed to this hit rate for observable cancers to be detected. The mechanism proposed is that the extensive prior DNA damage in smokers, followed by alpha particle damage to a critical site in checkpoint genes, accounts for the greater lung cancer response in smokers. PMID- 18420563 TI - Cost-effectiveness of countermeasures against radon in the Czech Republic: preliminary report. AB - The cost-effectiveness of remedial and preventive countermeasures against indoor radon in the frame of the Czech Radon Programme is preliminarily estimated, resulting in 72 700 CZK and 194 000 CZK per y of life gained by remediation and prevention, respectively, (exchange rate approximately 28 CZK/EUR). PMID- 18420564 TI - Radon in context of natural radiation exposure: the Czech experience. AB - Radon issue should be seen in the context of other human exposures and harms. The range of doses from the internal (inhalation and ingestion) and external exposure pathways is presented. Indoor radon regulation is compared with regulation of other natural exposures. PMID- 18420565 TI - A preliminary radon map for Canada according to health region. AB - The recent publications of the combined analyses of residential radon studies in Europe and North America have shown that there is a significant risk of lung cancer at residential radon levels. In order to assess the population risk due to radon, the knowledge of the spatial distribution of indoor radon levels is essential. Here a preliminary radon map for Canada is presented, based on historical radon measurements collected in 6016 locations across Canada with the health region as the basic geographic units. PMID- 18420566 TI - Radiation dose from a linear slit scanning X-ray machine with full-body imaging capabilities. AB - Doses for a range of examinations and views using digital X-ray equipment with full-body linear slit scanning capabilities (Statscan) have been compared with those from other published studies. Entrance doses (free-in-air) were measured using a dosimeter, and effective doses were generated using a Monte Carlo simulator. Doses delivered by the linear slit scanning system were significantly lower than those from conventional X-ray equipment. Effective doses were between 9 and 75% of the United Nations Scientific Committee Report on the Effects of Ionising Radiation doses for standard examinations. This dose reduction can be explained by the properties of linear slit scanning technology, including low scatter, beam geometry, the use of a digital detector and the use of higher than usual tube voltages. PMID- 18420567 TI - Proposed preliminary diagnostic reference levels for three common interventional cardiology procedures in Ireland. AB - This study has gathered data across Ireland to determine the range of radiation doses received during interventional cardiology (IC) investigations. Radiation doses for three common types of IC examinations where investigated: coronary angiography (CA), percutaneous coronary intervention (PCI) and pacemaker insertions (PPI). A total of 22 cardiac imaging suites participated in the study. Radiation dose was monitored for 1804 adult patients using dose area product (DAP) meters. Individual patient DAP values ranged from 136-23,101 cGy cm2, 475 41,038 cGy cm2 and 45-17,192 cGy cm2 for CA, PCI and PPI respectively, with third quartile values of 4654 cGy cm2, 10,650 cGy cm2 and 1686 cGy cm2. The importance of optimising radiation dose, while not compromising diagnostic efficacy is clear. Although setting reference levels for these complex procedures has some difficulties, it is important that some guideline values are available as a benchmark to guide the operators during these potentially high dose procedures. The third quartile values as described by this paper may offer such guidance. PMID- 18420568 TI - Applying the Ionising Radiation Regulations to radon in the UK workplace. AB - As a response to the identification of a health risk from workplace radon in the UK, the Ionising Radiations Regulations include the protection of workers from excessive levels of radon. Employers are required to make risk assessments, and the interpretation of the Health and Safety Executive is that the regulations apply to workplace premises in locations already designated as Radon Affected Areas for domestic purposes, with the difference that in workplaces, it is the maximum winter radon concentration rather than the annual average which is the parameter of interest. This paper discusses the rationale behind the current regulatory environment, outlines the role and duties of Accredited Radiation Protection Advisers and summarises the strategies necessary to conform to the regulations. PMID- 18420569 TI - Factors affecting the transfer of radionuclides from the environment to plants. AB - Much of our food directly or indirectly originates from plant material; thus, detailed studies on plant contamination processes are an essential part of international environmental research. This overview attempts to identify and describe the most important parameters and processes affecting the behaviour of radionuclide transfer to plants. Many parameters influence these processes. These parameters are related to: (1) plant, (2) soil, (3) radionuclide, (4) climate and (5) time. Often there is no boundary between the factors and they are linked to each other. Knowledge of important factors in radionuclide transfer to plants can help to assess and prevent radiological exposure of humans. This knowledge can also help to guide researches and modelling related to transfer of radionuclides to food chain. PMID- 18420571 TI - Comments on 'The thermoluminescence dose-response and other characteristics of the high-temperature TL in LiF: Mg.Ti (TLD-100)' by Y.S. Horowitz, L. Oster and H. Datz. PMID- 18420572 TI - New concept of IEC standards for radiation protection dosemeters. AB - The International Electrotechnical Commission (IEC) develops new standards for radiation protection dosemeters which follow a new concept. They are much more flexible in detail, but still ensure the same measurement quality. They are, for example, no longer specific for the detector type, but rather specific for the measurement task, e.g. for individual monitoring with active direct-reading instruments. Another example is that they are flexible with respect to the ranges of influence quantities. The conceptual changes are described in this paper, together with the advantages this new concept provides for manufacturers, users and legislators. PMID- 18420573 TI - An empirical multivariate log-normal distribution representing uncertainty of biokinetic parameters for 137Cs. AB - A simplified biokinetic model for (137)Cs has six parameters representing transfer of material to and from various compartments. Using a Bayesian analysis, the joint probability distribution of these six parameters is determined empirically for two cases with quite a lot of bioassay data. The distribution is found to be a multivariate log-normal. Correlations between different parameters are obtained. The method utilises a fairly large number of pre-determined forward biokinetic calculations, whose results are stored in interpolation tables. Four different methods to sample the multidimensional parameter space with a limited number of samples are investigated: random, stratified, Latin Hypercube sampling with a uniform distribution of parameters and importance sampling using a lognormal distribution that approximates the posterior distribution. The importance sampling method gives much smaller sampling uncertainty. No sampling method-dependent differences are perceptible for the uniform distribution methods. PMID- 18420574 TI - Radon, thoron and their decay products in natural caves with nuclear track detectors. AB - Alpha and beta activities per unit volume of air due to radon (222Rn), thoron (220Rn) and their progenies were measured in the air of natural caves by using CR 39 and LR-115 type II solid-state nuclear track detectors. Equilibrium factors between radon and its daughters and between thoron and its progeny were evaluated in the atmospheres studied. PMID- 18420575 TI - Radon programme: presence and future. AB - An overview of radon programme experience is presented. The paper summarises broad topics concerning radon issue: philosophy of radon policy, radon measurements strategies with respect to indoor radon variation, progress in radon measurement of an individual house (radon diagnosis), national programmes, the role of preventive measures and interventions with respect to existing and future exposure and knowledge of radon risk, problems of remediation strategies, radon mapping process and sense of delineation of radon prone areas, public awareness on radon issue and publicity campaign. Some research activities are proposed aiming at effective solutions for radon issues in the future. PMID- 18420576 TI - Biological roles for the NOX family NADPH oxidases. PMID- 18420577 TI - O-GlcNAc regulates FoxO activation in response to glucose. AB - FoxO proteins are key transcriptional regulators of nutrient homeostasis and stress response. The transcription factor FoxO1 activates expression of gluconeogenic, including phosphoenolpyruvate carboxykinase and glucose-6 phosphatase, and also activates the expression of the oxidative stress response enzymes catalase and manganese superoxide dismutase. Hormonal and stress dependent regulation of FoxO1 via acetylation, ubiquitination, and phosphorylation, are well established, but FoxOs have not been studied in the context of the glucose-derived O-linked beta-N-acetylglucosamine (O-GlcNAc) modification. Here we show that O-GlcNAc on hepatic FoxO1 is increased in diabetes. Furthermore, O-GlcNAc regulates FoxO1 activation in response to glucose, resulting in the paradoxically increased expression of gluconeogenic genes while concomitantly inducing expression of genes encoding enzymes that detoxify reactive oxygen species. GlcNAcylation of FoxO provides a new mechanism for direct nutrient control of transcription to regulate metabolism and stress response through control of FoxO1 activity. PMID- 18420578 TI - Leishmania trypanothione synthetase-amidase structure reveals a basis for regulation of conflicting synthetic and hydrolytic activities. AB - The bifunctional trypanothione synthetase-amidase catalyzes biosynthesis and hydrolysis of the glutathione-spermidine adduct trypanothione, the principal intracellular thiol-redox metabolite in parasitic trypanosomatids. These parasites are unique with regard to their reliance on trypanothione to determine intracellular thiol-redox balance in defense against oxidative and chemical stress and to regulate polyamine levels. Enzymes involved in trypanothione biosynthesis provide essential biological activities, and those absent from humans or for which orthologues are sufficiently distinct are attractive targets to underpin anti-parasitic drug discovery. The structure of Leishmania major trypanothione synthetase-amidase, determined in three crystal forms, reveals two catalytic domains. The N-terminal domain, a cysteine, histidine-dependent amidohydrolase/peptidase amidase, is a papain-like cysteine protease, and the C terminal synthetase domain displays an ATP-grasp family fold common to C:N ligases. Modeling of substrates into each active site provides insight into the specificity and reactivity of this unusual enzyme, which is able to catalyze four reactions. The domain orientation is distinct from that observed in a related bacterial glutathionylspermidine synthetase. In trypanothione synthetase-amidase, the interactions formed by the C terminus, binding in and restricting access to the amidase active site, suggest that the balance of ligation and hydrolytic activity is directly influenced by the alignment of the domains with respect to each other and implicate conformational changes with amidase activity. The potential inhibitory role of the C terminus provides a mechanism to control relative levels of the critical metabolites, trypanothione, glutathionylspermidine, and spermidine in Leishmania. PMID- 18420579 TI - Store-dependent and -independent modes regulating Ca2+ release-activated Ca2+ channel activity of human Orai1 and Orai3. AB - We evaluated currents induced by expression of human homologs of Orai together with STIM1 in human embryonic kidney cells. When co-expressed with STIM1, Orai1 induced a large inwardly rectifying Ca(2+)-selective current with Ca(2+)-induced slow inactivation. A point mutation of Orai1 (E106D) altered the ion selectivity of the induced Ca(2+) release-activated Ca(2+) (CRAC)-like current while retaining an inwardly rectifying I-V characteristic. Expression of the C-terminal portion of STIM1 with Orai1 was sufficient to generate CRAC current without store depletion. 2-APB activated a large relatively nonselective current in STIM1 and Orai3 co-expressing cells. 2-APB also induced Ca(2+) influx in Orai3-expressing cells without store depletion or co-expression of STIM1. The Orai3 current induced by 2-APB exhibited outward rectification and an inward component representing a mixed calcium and monovalent current. A pore mutant of Orai3 inhibited store-operated Ca(2+) entry and did not carry significant current in response to either store depletion or addition of 2-APB. Analysis of a series of Orai1-3 chimeras revealed the structural determinant responsible for 2-APB induced current within the sequence from the second to third transmembrane segment of Orai3. The Orai3 current induced by 2-APB may reflect a store independent mode of CRAC channel activation that opens a relatively nonselective cation pore. PMID- 18420580 TI - Targeted ablation of glucose-dependent insulinotropic polypeptide-producing cells in transgenic mice reduces obesity and insulin resistance induced by a high fat diet. AB - The K cell is a specific sub-type of enteroendocrine cell located in the proximal small intestine that produces glucose-dependent insulinotropic polypeptide (GIP), xenin, and potentially other unknown hormones. Because GIP promotes weight gain and insulin resistance, reducing hormone release from K cells could lead to weight loss and increased insulin sensitivity. However, the consequences of coordinately reducing circulating levels of all K cell-derived hormones are unknown. To reduce the number of functioning K cells, regulatory elements from the rat GIP promoter/gene were used to express an attenuated diphtheria toxin A chain in transgenic mice. K cell number, GIP transcripts, and plasma GIP levels were profoundly reduced in the GIP/DT transgenic mice. Other enteroendocrine cell types were not ablated. Food intake, body weight, and blood glucose levels in response to insulin or intraperitoneal glucose were similar in control and GIP/DT mice fed standard chow. In contrast to single or double incretin receptor knock out mice, the incretin response was absent in GIP/DT animals suggesting K cells produce GIP plus an additional incretin hormone. Following high fat feeding for 21-35 weeks, the incretin response was partially restored in GIP/DT mice. Transgenic versus wild-type mice demonstrated significantly reduced body weight (25%), plasma leptin levels (77%), and daily food intake (16%) plus enhanced energy expenditure (10%) and insulin sensitivity. Regardless of diet, long term glucose homeostasis was not grossly perturbed in the transgenic animals. In conclusion, studies using GIP/DT mice demonstrate an important role for K cells in the regulation of body weight and insulin sensitivity. PMID- 18420581 TI - N-linked glycosylation selectively regulates the generic folding of HLA-Cw1. AB - To resolve primary (glycosylation-assisted) from secondary (glycosylation independent) quality control steps in the biosynthesis of HLA (human leukocyte antigen) class I glycoproteins, the unique N-linked glycosylation site of the HLA Cw1 heavy chain was deleted by site-directed mutagenesis. The non-glycosylated Cw1S88G mutant was characterized by flow cytometry, pulse-chase, co immunoprecipitation, and in vitro assembly assays with synthetic peptide ligands upon transfection in 721.221 and 721.220 cells. The former provide a full set of primary as well as secondary chaperoning interactions, whereas the latter are unable to perform secondary quality control (e.g. proper class I assembly with peptide antigens) as a result of a functional defect of the HLA-dedicated chaperone tapasin. In both transfectants, Cw1S88G displayed a loss/weakening in its generic chaperoning interaction with calreticulin and/or ERp57 and became redistributed toward calnexin, known to bind the most unfolded class I conformers. Despite this, and quite unexpectedly, a weak interaction with the HLA dedicated chaperone TAP was selectively retained in 721.221. In addition, the ordered, stepwise acquisition of thermal stability/peptide binding was disrupted, resulting in a heterogeneous ensemble of Cw1S88G conformers with unorthodox and unprecedented peptide assembly features. Because a lack of glycosylation and a lack of tapasin-assisted peptide loading have distinct, complementary, and additive effects, the former is separable from (and upstream of) the latter, e.g. primary quality control is suggested to supervise a crucial, generic folding step preliminary to the acquisition of peptide receptivity. PMID- 18420582 TI - The hairless phenotype of the Hirosaki hairless rat is due to the deletion of an 80-kb genomic DNA containing five basic keratin genes. AB - Most models of hereditary hypotrichosis are due to alterations in growth factors and transcription factors, and the examples of causative mutations in hair keratin genes are limited. The Hirosaki hairless rat (HHR) is a mutant strain spontaneously derived from Sprague-Dawley rats (SDRs). In this study, the locus of the responsible gene was examined by linkage analysis and mapped on chromosome 7q36. Because many basic keratin genes are clustered on 7q36, their expression was examined. Reverse transcription-PCR and genomic PCR indicated that the Kb21 (Krt81), -23 (Krt83), and -26 (Krt86) genes encoding basic hair keratins were not expressed and were deleted. Furthermore, 80-kb genomic DNA ranging from exon 9 of Kb25 (Krt85) to exon 9 of Krt2-25 was deleted. The breakpoints of these genes were within a 95-bp portion shared by the two genes, suggesting that deletion due to non-allelic homologous recombination occurred. Proteins identified as Kb21, Kb23, and Krt2-25 in SDR hairs by mass spectrometry were not detected in HHR. Instead, the product of a fusion gene became dominant in HHR. Because fusion occurred between the exons of the two genes with the same sequences, the product was identical to the wild-type Kb25 protein. By using immunohistochemistry, Kb21 was not detected in HHR hair follicles. Kb25 was expressed in the cortex in HHRs, whereas it was in the medulla in SDRs. This study clearly illustrates the importance of hair keratin genes in hair growth. PMID- 18420583 TI - Insulin-like growth factor-I stimulates Shc-dependent phosphatidylinositol 3 kinase activation via Grb2-associated p85 in vascular smooth muscle cells. AB - Insulin-like growth factor-I (IGF-I) stimulates vascular smooth muscle cell proliferation and migration by activating both MAPK and phosphatidylinositol 3 kinase (PI3K). Vascular smooth muscle cells (VSMCs) maintained in 25 mm glucose sustain MAPK activation via increased Shc phosphorylation and Grb2 association resulting in an enhanced mitogenic response compared with cells grown in 5 mm glucose. PI3K plays a major role in IGF-I-stimulated VSMC migration, and hyperglycemia augments this response. In contrast to MAPK activation the role of Shc in modulating PI3K in response to IGF-I has not been determined. In this study we show that impaired Shc association with Grb2 results in decreased Grb2 p85 association, SHPS-1-p85 recruitment, and PI3K activation in response to IGF I. Exposure of VSMCs to cell-permeable peptides, which contained polyproline sequences from p85 proposed to mediate Grb2 association, resulted in inhibition of Grb2-p85 binding and AKT phosphorylation. Transfected cells that expressed p85 mutant that had specific prolines mutated to alanines resulted in less Grb2-p85 association, and a Grb2 mutant (W36A/W193A) that attenuated p85 binding showed decreased association of p85 with SHPS-1, PI3K activation, AKT phosphorylation, cell proliferation, and migration in response to IGF-I. Cellular exposure to 25 mm glucose, which is required for Shc phosphorylation in response to IGF-I, resulted in enhanced Grb2 binding to p85, activation of PI3K activity, and increased AKT phosphorylation as compared with cells exposed to 5 mm glucose. We conclude that in VSMCs exposed to hyperglycemia, IGF-I stimulation of Shc facilitates the transfer of Grb2 to p85 resulting in enhanced PI3K activation and AKT phosphorylation leading to enhanced cell proliferation and migration. PMID- 18420584 TI - Translation regulation by eukaryotic initiation factor-2 kinases in the development of latent cysts in Toxoplasma gondii. AB - A key problem in the treatment of numerous pathogenic eukaryotes centers on their development into latent forms during stress. For example, the opportunistic protist Toxoplasma gondii converts to latent cysts (bradyzoites) responsible for recrudescence of disease. We report that Toxoplasma eukaryotic initiation factor 2alpha (TgIF2alpha) is phosphorylated during stress and establish that protozoan parasites utilize translation control to modulate gene expression during development. Importantly, TgIF2alpha remains phosphorylated in bradyzoites, explaining how these cells maintain their quiescent state. Furthermore, we have characterized novel eIF2 kinases; one in the endoplasmic reticulum and a likely regulator of the unfolded protein response (TgIF2K-A) and another that is a probable responder to cytoplasmic stresses (TgIF2K-B). Significantly, our data suggest that 1) the regulation of protein translation through eIF2 kinases is associated with development, 2) eIF2alpha phosphorylation is employed by cells to maintain a latent state, and 3) endoplasmic reticulum and cytoplasmic stress responses evolved in eukaryotic cells before the early diverging Apicomplexa. Given its importance to pathogenesis, eIF2 kinase-mediated stress responses may provide opportunities for novel therapeutics. PMID- 18420585 TI - RACK1 and CIS mediate the degradation of BimEL in cancer cells. AB - RACK1 is a 7-WD motif-containing protein with numerous downstream effectors regulating various cellular functions. Using a yeast two-hybrid screen, we identified dynein light chain 1 as a novel interacting partner of RACK1. Additionally, we demonstrated that RACK1 formed a complex with DLC1 and Bim, specifically BimEL, in the presence of apoptotic agents. Upon paclitaxel treatment, RACK1, DLC1, and CIS mediated the degradation of BimEL through the ElonginB/C-Cullin2-CIS ubiquitin-protein isopeptide ligase complex. We further showed that RACK1 conferred paclitaxel resistance to breast cancer cells in vitro and in vivo. Finally, we observed an inverse correlation between CIS and BimEL levels in both ovarian and breast cancer cell lines and specimens. Our study suggests a role of RACK1 in protecting cancer cells from apoptosis by regulating the degradation of BimEL, which together with CIS could play an important role of drug resistance in chemotherapy. PMID- 18420586 TI - Caspase cleavage of HER-2 releases a Bad-like cell death effector. AB - Human epidermal growth factor receptor-2 (HER-2/ErbB2/neu), a receptor tyrosine kinase that is amplified/overexpressed in poor prognosis breast carcinomas, confers resistance to apoptosis by activating cell survival pathways. Here we demonstrate that the cytoplasmic tail of HER-2 is cleaved by caspases at Asp(1016)/Asp(1019) to release a approximately 47-kDa product, which is subsequently proteolyzed by caspases at Asp(1125) into an unstable 22-kDa fragment that is degraded by the proteasome and a predicted 25-kDa product. Both the 47- and 25-kDa products translocate to mitochondria, release cytochrome c by a Bcl-x(L)-suppressible mechanism, and induce caspase-dependent apoptosis. The 47 and 25-kDa HER-2 cleavage products share a functional BH3-like domain, which is required for cytochrome c release in cells and isolated mitochondria and for apoptosis induction. Caspase-cleaved HER-2 binds Bcl-x(L) and acts synergistically with truncated Bid to induce apoptosis, mimicking the actions of the BH3-only protein Bad. Moreover, the HER-2 cleavage products cooperate with Noxa to induce apoptosis in cells expressing both Bcl-x(L) and Mcl-1, confirming their Bad-like function. Collectively, our results indicate that caspases activate a previously unrecognized proapoptotic function of HER-2 by releasing a Bad-like cell death effector. PMID- 18420587 TI - Ribosomal protein S9 is a novel B23/NPM-binding protein required for normal cell proliferation. AB - B23 (NPM/nucleophosmin) is a multifunctional nucleolar protein and a member of the nucleoplasmin superfamily of acidic histone chaperones. B23 is essential for normal embryonic development and plays an important role in genomic stability, ribosome biogenesis, and anti-apoptotic signaling. Altered protein expression or genomic mutation of B23 is encountered in many different forms of cancer. Although described as multifunctional, a genuine molecular function of B23 is not fully understood. Here we show that B23 is associated with a protein complex consisting of ribosomal proteins and ribosome-associated RNA helicases. A novel, RNA-independent interaction between ribosomal protein S9 (RPS9) and B23 was further investigated. We found that S9 binding requires an intact B23 oligomerization domain. Depletion of S9 by small interfering RNA resulted in decreased protein synthesis and G(1) cell cycle arrest, in association with induction of p53 target genes. We determined that S9 is a short-lived protein in the absence of ribosome biogenesis, and proteasomal inhibition significantly increased S9 protein level. Overexpression of B23 facilitated nucleolar storage of S9, whereas knockdown of B23 led to diminished levels of nucleolar S9. Our results suggest that B23 selectively stores, and protects ribosomal protein S9 in nucleoli and therefore could facilitate ribosome biogenesis. PMID- 18420588 TI - The role of CDC48 in the retro-translocation of non-ubiquitinated toxin substrates in plant cells. AB - When the catalytic A subunits of the castor bean toxins ricin and Ricinus communis agglutinin (denoted as RTA and RCA A, respectively) are delivered into the endoplasmic reticulum (ER) of tobacco protoplasts, they become substrates for ER-associated protein degradation (ERAD). As such, these orphan polypeptides are retro-translocated to the cytosol, where a significant proportion of each protein is degraded by proteasomes. Here we begin to characterize the ERAD pathway in plant cells, showing that retro-translocation of these lysine-deficient glycoproteins requires the ATPase activity of cytosolic CDC48. Lysine polyubiquitination is not obligatory for this step. We also show that although RCA A is found in a mannose-untrimmed form prior to its retro-translocation, a significant proportion of newly synthesized RTA cycles via the Golgi and becomes modified by downstream glycosylation enzymes. Despite these differences, both proteins are similarly retro-translocated. PMID- 18420589 TI - G-protein-coupled receptor-mediated traffic of Na,K-ATPase to the plasma membrane requires the binding of adaptor protein 1 to a Tyr-255-based sequence in the alpha-subunit. AB - Motion of integral membrane proteins to the plasma membrane in response to G protein-coupled receptor signals requires selective cargo recognition motifs that bind adaptor protein 1 and clathrin. Angiotensin II, through the activation of AT1 receptors, promotes the recruitment to the plasma membrane of Na,K-ATPase molecules from intracellular compartments. We present evidence to demonstrate that a tyrosine-based sequence (IVVY-255) present within the Na,K-ATPase alpha1 subunit is involved in the binding of adaptor protein 1. Mutation of Tyr-255 to a phenylalanine residue in the Na,K-ATPase alpha1-subunit greatly reduces the angiotensin II-dependent activation of Na,K-ATPase, recruitment of Na,K-ATPase molecules to the plasma membrane, and association of adaptor protein 1 with Na,K ATPase alpha1-subunit molecules. To determine protein-protein interaction, we used fluorescence resonance energy transfer between fluorophores attached to the Na,K-ATPase alpha1-subunit and adaptor protein 1. Although angiotensin II activation of AT1 receptors induces a significant increase in the level of fluorescence resonance energy transfer between the two molecules, this effect was blunted in cells expressing the Tyr-255 mutant. Thus, results from different methods and techniques suggest that the Tyr-255-based sequence within the NKA alpha1-subunit is the site of adaptor protein 1 binding in response to the G protein-coupled receptor signals produced by angiotensin II binding to AT1 receptors. PMID- 18420590 TI - Assistance for a chaperone: Chlamydomonas HEP2 activates plastidic HSP70B for cochaperone binding. AB - Previous efforts aimed at the biochemical characterization of chloroplast HSP70B were hampered by the observation that recombinant HSP70B was inactive, i.e. incompetent of interacting with its nucleotide exchange factor CGE1. In addition, because heterologously expressed mitochondrial Hsp70 was inactive unless coexpressed with the escort protein Hep1, we wondered whether homologs of Hep1 existed in the chloroplast. Data base searches revealed that algae and higher plants indeed encode at least two HEP homologs, one predicted to be targeted to mitochondria, the others to chloroplasts. Using Chlamydomonas reinhardtii as plant model organism we demonstrate that this alga encodes an HEP homolog (termed HEP2) that is localized to the stroma. HEP2 is expressed constitutively as a low abundance protein with an apparent molecular mass of approximately 21 kDa. In cell extracts HEP2 interacts with HSP70B in an ATP-dependent fashion. Coexpression of HSP70B with HEP2 in Escherichia coli yielded high levels of CGE1 binding competent HSP70B, which also displayed ATPase activity. Inactive HSP70B was more prone to proteolysis than active HSP70B. Although inactive HSP70B interacted with HEP2, it could not be activated. Active HSP70B remained active for 48 h in the absence of HEP2, suggesting that HEP2 was not involved in maintaining HSP70B in an active state. However, some HSP70B expressed as a fusion protein with an N-terminal extension was activated when HEP2 was present during cleavage of the fusion protein, suggesting that in vivo HEP2 might be required for de novo folding of HSP70B after transit peptide cleavage. PMID- 18420591 TI - Root hydraulic conductivity measured by pressure clamp is substantially affected by internal unstirred layers. AB - Using the root pressure probe in the pressure clamping (PC) mode, the impact of internal unstirred layers (USLs) was quantified for young corn roots, both in experiments and in computer simulations applying the convection/diffusion model of Knipfer et al. In the experiments, water flows (J(Vr)s) during PC were analysed in great detail, showing that J(Vr)s (and the apparent root hydraulic conductivity) were high during early stages of PC and declined rapidly during the first 80 s of clamping to a steady-state value of 40-30% of the original. The comparison of experimental results with simulations showed that, during PC, internal USLs at the inner surface of the endodermis substantially modify the overall force driving the water. As a consequence, J(Vr) and Lp(r) were inhibited. Effects of internal USLs were minimized when using the pressure relaxation mode, when internal USLs had not yet developed. Additional stop-clamp experiments and experiments where the endodermis was punctured to reduce the effect of internal USLs verified the existence of internal USLs during PC. Data indicated that the role of pressure propagation along the root xylem for both PC and pressure relaxation modes should be small, as should the effects of filling of the capacities during root pressure probe experiments, which are discussed as an alternative model. The results supported the idea that concentration polarization effects at the endodermis (internal USLs) cause a serious problem whenever relatively large amounts of water (xylem sap) are radially moved across the root, such as during PC or when using the high-pressure flow meter technique. PMID- 18420592 TI - Molecular cloning and characterization of a moss (Ceratodon purpureus) nonsymbiotic hemoglobin provides insight into the early evolution of plant nonsymbiotic hemoglobins. AB - Nonsymbiotic hemoglobins (nsHbs) are widespread in plants including bryophytes. Bryophytes (such as mosses) are among the oldest land plants, thus an analysis of a bryophyte nsHb is of interest from an evolutionary perspective. However, very little is known about bryophyte nsHbs. Here, we report the cloning and characterization of an nshb gene (cerhb) from the moss Ceratodon purpureus. Sequence analysis showed that cerhb is interrupted by 3 introns in identical position as all known plant nshb genes, which suggests that the ancestral nshb gene was interrupted by 3 introns. Expression analysis showed that cerhb expresses in protonemas and gametophytes growing in normal conditions and that it overexpresses in protonemas subjected to osmotic (sucrose), heat-shock, cold-, and nitrate-stress conditions. Also, modeling of the Ceratodon nsHb (CerHb) tertiary structure suggests that CerHb is hexacoordinate and that it binds O(2) with high affinity. Comparative analysis of the predicted CerHb with native rice Hb1 and soybean leghemoglobin a structures revealed that the major evolutionary changes that probably occurred during the evolution of plant Hbs were 1) a hexacoordinate to pentacoordinate transition at the heme prosthetic group, 2) a length decrease at the CD-loop and N- and C-termini regions, and 3) the compaction of the protein into a globular structure. PMID- 18420593 TI - Bayesian inference of errors in ancient DNA caused by postmortem degradation. AB - Methods for extracting and amplifying sequences using ancient DNA (aDNA) can be prone to errors caused by postmortem modifications of the DNA strand. A new statistical method is developed for predicting errors in aDNA sequences caused by such processes. In addition to the canonical DNA substitution model parameters, a discrete Markov chain is used to describe nucleotide substitutions occurring via postmortem degradation of the aDNA sequences. A computer program, BYPASSR-degr, was developed implementing the method and was used in subsequent analyses of simulated data sets under the new model. Simulation studies show that the new method can be powerful and accurate in identifying damaged sites. The method is applied to analyze aDNA sequences of Etruscans, Adelie penguins, and horses. No significant signals of degradation were observed at any sites of the aDNA sequences we analyzed. PMID- 18420594 TI - The gravity-regulated growth of axillary buds is mediated by a mechanism different from decapitation-induced release. AB - When the upper part of the main shoot of the Japanese morning glory (Pharbitis nil or Ipomoea nil) is bent down, the axillary bud situated on the uppermost node of the bending region is released from apical dominance and elongates. Here, we demonstrate that this release of axillary buds from apical dominance is gravity regulated. We utilized two agravitropic mutants of morning glory defective in gravisensing cell differentiation, weeping (we) and weeping2 (we2). Bending the main shoots of either we or we2 plants resulted in minimal elongation of their axillary buds. This aberration was genetically linked to the agravitropism phenotype of the mutants, which implied that shoot bending-induced release from apical dominance required gravisensing cells. Previous studies have shown that basipetal translocation of auxin from the apical bud inhibits axillary bud growth, whereas cytokinin promotes axillary bud outgrowth. We therefore compared the roles of auxin and cytokinin in bending- or decapitation-induced axillary bud growth. In the wild-type and we plants, decapitation increased cytokinin levels and reduced auxin response. In contrast, shoot bending did not cause significant changes in either cytokinin level or auxin response, suggesting that the mechanisms underlying gravity- and decapitation-regulated release from apical dominance are distinct and unique. PMID- 18420595 TI - Overexpression of a new rice vacuolar antiporter regulating protein OsARP improves salt tolerance in tobacco. AB - We examined the function of the rice (Oryza sativa L.) antiporter-regulating protein OsARP by overexpressing it in tobacco (Nicotiana tabacum L.). In public databases, this protein was annotated as a putative Os02g0465900 protein of rice. The OsARP gene was introduced into tobacco under the control of the cauliflower mosaic virus 35S promoter. The transformants were selected for their ability to grow on medium containing kanamycin. Incorporation of the transgene in the genome of tobacco was confirmed by PCR, and its expression was confirmed by Western blot analysis. Transgenic plants had better growth and vigor than non-transgenic plants under salt stress in vitro. Overexpression of OsARP in transgenic tobacco plants resulted in salt tolerance, and the plants had a higher rate of photosynthesis and effective PSII photon yield when compared with the wild type. The OsARP protein was localized in the tonoplast of rice plants. Transgenic plants accumulated more Na+ in their leaf tissue than did wild-type plants. It is conceivable that the toxic effect of Na+ in the cytosol might be reduced by sequestration into vacuoles. The rate of water loss was higher in the wild type than in transgenic plants under salt stress. Increased vacuolar solute accumulation and water retention could confer salt tolerance in transgenic plants. Tonoplast vesicles isolated from OsARP transgenic plants showed Na+/H+ exchange rates 3-fold higher than those of wild-type plants. These results suggest that OsARP on the tonoplasts plays an important role in compartmentation of Na+ into vacuoles. We suggest that OsARP is a new type of protein participating in Na+ uptake in vacuoles. PMID- 18420596 TI - Cytokinin inhibits the proteasome-mediated degradation of carbonylated proteins in Arabidopsis leaves. AB - Under normal conditions, plants contain numerous carbonylated proteins, which are thought to be indicative of oxidative stress damage. Conditions that promote formation of reactive oxygen species (ROS) enhance protein carbonylation, and protein degradation is required to reverse the damage. However, it is not clear how the degradation of carbonylated proteins is controlled in planta. In this report, we show that detached Arabidopsis leaves rapidly and selectively degrade carbonylated proteins when kept in the dark. The loss of carbonylated proteins corresponded to a loss of soluble protein and accumulation of free amino acids. Degradation of carbonylated proteins and the loss of soluble protein was blocked by MG132 but not 3-methyladenine, suggesting that the 26S proteasome pathway rather than the autophagic pathway was involved. Consistent with this, rpn10 and rpn12 mutants, which are defective in proteasome function, had increased (rather than decreased) levels of carbonylated proteins when detached in the dark. Feeding metabolites (amino acids and sucrose) to detached leaves of wild-type Arabidopsis in the dark had little or no effect on the loss of carbonylated proteins, whereas providing soybean xylem sap via the transpiration stream effectively prevented degradation. The effect of xylem sap was mimicked by feeding 10 muM kinetin. We postulate that disruption of cytokinin flux to detached leaves triggers the selective degradation of carbonylated proteins via the proteasome pathway. The results may have implications for the control of protein mobilization in response to changes in N availability. PMID- 18420597 TI - Requirement of Ala residues at g position in heptad sequence of alpha-helix forming peptide for formation of fibrous structure. AB - One feature of the alpha3-peptide, which has the amino acid sequence of (Leu-Glu Thr-Leu-Ala-Lys-Ala)(3), that distinguishes it from many other alpha-helix forming peptides is its ability to form fibrous assemblies that can be observed by transmission electron microscopy. In this study, the effects of Ala-->Gln substitution at the e (5th) or g (7th) position in the above heptad sequence of the alpha3-peptide on the formation of alpha-helix and fibrous assemblies were investigated by circular dichroism spectral measurement and atomic force microscopy. The 5Qalpha3-peptide obtained by Ala-->Gln substitution at the e position of the alpha3-peptide was found to form very short fibrils with long elliptical shape, whereas the 7Qalpha3-peptide with Gln residues at the g position lost its ability to form such assemblies, in spite of alpha-helix formation in both peptides; the stabilities of both peptides decreased. These results indicate that Ala residues at the g position in the heptad sequence of the alpha3-peptide are key residues for the formation of fibrous assemblies, which may be due to hydrophobic interactions between alpha-helical bundle surfaces. PMID- 18420598 TI - Chemical and apoptotic properties of hydroxy-ceramides containing long-chain bases with unusual alkyl chain lengths. AB - We analysed four types of free ceramides (Cer 1, Cer 2, Cer 3 and Cer 4) from equine kidneys by electrospray ionization mass spectrometry. Cer 1 was composed of dihydroxy long-chain bases (dLCBs) of (4E)-sphingenine (d18:1), sphinganine and non-hydroxy fatty acids (NFAs); Cer 2 was composed of trihydroxy LCBs (tLCBs) of 4-hydroxysphinganine, t16:0, t18:0, t19:0 and t20:0, and NFAs; Cer 3 was composed of dLCBs, d16:1, d17:1, d18:1, d19:1 and d20:1, and hydroxy FAs (HFAs); and Cer 4 was composed of tLCBs, t16:0, t17:0, t18:0, t19:0 and t20:0, and HFAs. The results indicate all ceramide species containing LCBs with non-octadeca lengths (NOD-LCBs) can be classified into hydroxy-ceramides since these species always consist of tLCBs, and/or HFAs. Furthermore, such species tend to contain FAs with longer acyl chains but contain neither palmitate (C16:0) nor its hydroxylated form (C16:0h). The apoptosis-inducing activities of these hydroxyl ceramides towards tumour cell lines were compared with that of non-hydroxy ceramides, dLCB-NFA (Cer 1). Monohydroxy-ceramides, tLCB-NFA (Cer 2) and dLCB-HFA (Cer 3), exhibited stronger activities, whereas dihydroxy-ceramides, tLCB-HFA (Cer 4), exhibited similar or weaker activity than dLCB-NFA (Cer 1), depending on cell lines. PMID- 18420599 TI - Antipsychotic-like properties of phosphodiesterase 4 inhibitors: evaluation of 4 (3-butoxy-4-methoxybenzyl)-2-imidazolidinone (RO-20-1724) with auditory event related potentials and prepulse inhibition of startle. AB - Antipsychotic medications function through antagonism of D2 dopamine receptors. Blockade of D2 receptors causes an increase in intracellular cAMP, a ubiquitous second messenger. Inhibition of phosphodiesterase (PDE) activity, a family of enzymes that degrade cyclic nucleotides, causes the same effect. The conceptual linkage between dopamine D2 receptors and PDE activity via cAMP suggests a possible therapeutic potential for PDE inhibitors in schizophrenia. The limited number of studies in support of this hypothesis used rolipram, a specific inhibitor of the PDE4 family. In this study, we investigated the impact of 4-(3 butoxy-4-methoxybenzyl)-2-imidazolidinone (RO-20-1724), another PDE4-specific inhibitor, on auditory event-related potentials (ERPs), prepulse inhibition (PPI) of the startle reflex, and locomotor activity in mice. The ability to reverse amphetamine-induced alterations in ERPs and PPI was used as a model for psychosis. ERPs after RO-20-1724 revealed increased amplitude for the P20 and N40 ERP components. RO-20-1724 reversed the disruptive effect of amphetamines on ERPs and restored gating at a dose that did not impair locomotor activity. However, RO 20-1724 failed to reverse a amphetamine-induced decrease of PPI. Inconsistent results between these two psychosis models suggest that pure sensory processing, as measured with auditory ERPs, may be more sensitive to the effects of intracellular cAMP than sensorimotor effects as assessed with PPI. It remains unclear whether antipsychotic-like properties are a common feature of PDE4 inhibition, or if they are restricted to the pharmacological profile of rolipram. Future studies should examine how PDE4 subtype specificity might contribute to differences between rolipram and RO-20-1724 in sensorimotor gating. PMID- 18420601 TI - Supramolecular structure of the casein micelle. AB - The supramolecular structure of colloidal casein micelles in milk was investigated by using a sample preparation protocol based on adsorption of proteins onto a poly-l-lysine and parlodion-coated copper grid, staining of proteins and calcium phosphate by uranyl oxalate, instantaneous freezing, and drying under a high vacuum. High-resolution transmission electron microscopy stereo-images were obtained showing the interior structure of casein micelles. On the basis of our interpretation of these images, an interlocked lattice model was developed in which both casein-calcium phosphate aggregates and casein polymer chains act together to maintain casein micelle integrity. The caseins form linear and branched chains (2 to 5 proteins long) interlocked by the casein-stabilized calcium phosphate nanoclusters. This model suggests that stabilization of calcium phosphate nanoclusters by phosphoserine domains of alpha(s1)-, alpha(s2)-, or beta-casein, or their combination, would orient their hydrophobic domains outward, allowing interaction and binding to other casein molecules. Other interactions between the caseins, such as calcium bridging, could also occur and further stabilize the supramolecule. The combination of having an interlocked lattice structure and multiple interactions results in an open, sponge-like colloidal supramolecule that is resistant to spatial changes and disintegration. Hydrophobic interactions between caseins surrounding a calcium phosphate nanocluster would prevent complete dissociation of casein micelles when the calcium phosphate nanoclusters are solubilized. Likewise, calcium bridging and other electrostatic interactions between caseins would prevent dissociation of the casein micelles into casein-calcium phosphate nanocluster aggregates when milk is cooled or urea is added to milk, and hydrophobic interactions are reduced. The appearance of both polymer chains and small aggregate particles during milk synthesis would also be expected based on this interlocked lattice model of casein micelles, and its supramolecule structure thus exhibits the principles of self-aggregation, interdependence, and diversity observed in nature. PMID- 18420600 TI - Antihyperalgesic effects of (R,E)-N-(2-hydroxy-2,3-dihydro-1H-inden-4-yl)-3-(2 (piperidin-1-yl)-4-(trifluoromethyl)phenyl)-acrylamide (AMG8562), a novel transient receptor potential vanilloid type 1 modulator that does not cause hyperthermia in rats. AB - Antagonists of the vanilloid receptor TRPV1 (transient receptor potential vanilloid type 1) have been reported to produce antihyperalgesic effects in animal models of pain. These antagonists, however, also caused concomitant hyperthermia in rodents, dogs, monkeys, and humans. Antagonist-induced hyperthermia was not observed in TRPV1 knockout mice, suggesting that the hyperthermic effect is exclusively mediated through TRPV1. Since antagonist induced hyperthermia is considered a hurdle for developing TRPV1 antagonists as therapeutics, we investigated the possibility of eliminating hyperthermia while maintaining antihyperalgesia. Here, we report four potent and selective TRPV1 modulators with unique in vitro pharmacology profiles (profiles A through D) and their respective effects on body temperature. We found that profile C modulator, (R,E)-N-(2-hydroxy-2,3-dihydro-1H-inden-4-yl)-3-(2-(piperidin-1-yl)-4 (trifluoromethyl)phenyl)acrylamide (AMG8562), blocks capsaicin activation of TRPV1, does not affect heat activation of TRPV1, potentiates pH 5 activation of TRPV1 in vitro, and does not cause hyperthermia in vivo in rats. We further profiled AMG8562 in an on-target (agonist) challenge model, rodent pain models, and tested for its side effects. We show that AMG8562 significantly blocks capsaicin-induced flinching behavior, produces statistically significant efficacy in complete Freund's adjuvant- and skin incision-induced thermal hyperalgesia, and acetic acid-induced writhing models, with no profound effects on locomotor activity. Based on the data shown here, we conclude that it is feasible to modulate TRPV1 in a manner that does not cause hyperthermia while maintaining efficacy in rodent pain models. PMID- 18420602 TI - Modeling of the effect of freezer conditions on the principal constituent parameters of ice cream by using response surface methodology. AB - A systematic analysis was carried out by using response surface methodology to create a quantitative model of the synergistic effects of conditions in a continuous freezer [mix flow rate (L/h), overrun (%), cylinder pressure (kPa), drawing temperature ( degrees C), and dasher speed (rpm)] on the principal constituent parameters of ice cream [rate of fat destabilization (%), mean air cell diameter (mum), and mean ice crystal diameter (mum)]. A central composite face-centered design was used for this study. Thirty-one combinations of the 5 above-mentioned freezer conditions were designed (including replicates at the center point), and ice cream samples were manufactured and examined in a continuous freezer under the selected conditions. The responses were the 3 variables given above. A quadratic model was constructed, with the freezer conditions as the independent variables and the ice cream characteristics as the dependent variables. The coefficients of determination (R(2)) were greater than 0.9 for all 3 responses, but Q(2), the index used here for the capability of the model for predicting future observed values of the responses, was negative for both the mean ice crystal diameter and the mean air cell diameter. Therefore, pruned models were constructed by removing terms that had contributed little to the prediction in the original model and by refitting the regression model. It was demonstrated that these pruned models provided good fits to the data in terms of R(2), Q(2), and ANOVA. The effects of freezer conditions were expressed quantitatively in terms of the 3 responses. The drawing temperature ( degrees C) was found to have a greater effect on ice cream characteristics than any of the other factors. PMID- 18420603 TI - Influence of lamb rennet paste containing probiotic on proteolysis and rheological properties of pecorino cheese. AB - Pecorino cheeses made from heat-treated ewes' milk using traditional lamb rennet paste (RP), lamb rennet paste containing Lactobacillus acidophilus (LA-5; RPL), and lamb rennet paste containing a mix of Bifidobacterium lactis (BB-12) and Bifidobacterium longum (BB-46; RPB) were characterized for proteolytic and rheological features during ripening. Consumer acceptance of cheeses at 60 d of ripening was evaluated. Lactobacillus acidophilus and Bifidobacterium mix displayed counts of 8 log(10) cfu/g and 9 log(10) cfu/g, respectively, in cheese during ripening. The RPB cheese displayed a greater degradation of casein (CN) matrix carried out by the enzymes associated to both Bifidobacterium mix and endogenous lactic acid microflora, resulting in the highest values of non-CN N and water-soluble N and the highest amount of alpha(s)-CN degradation products in cheese at 60 d of ripening. The RPL cheese displayed intermediate levels of lactic acid bacteria and of N fractions. The percentage of gamma-CN in RP and RPL cheeses at 60 d was 2-fold higher than in the cheese curd of the same groups, whereas the mentioned parameter was 3-fold higher in RPB cheese than in the corresponding fresh curd according to its highest plasmin content. The lower hardness in RPB at the end of ripening could be ascribed to the greater proteolysis observed in cheese harboring the Bifidobacterium mix. Although differences in proteolytic patterns were found among treatments, there were no differences in smell and taste scores. PMID- 18420604 TI - Microbial and sensory changes throughout the ripening of Prato cheese made from milk with different levels of somatic cells. AB - The objective of this research was to evaluate the effects of 2 levels of raw milk somatic cell count (SCC) on the composition of Prato cheese and on the microbiological and sensory changes of Prato cheese throughout ripening. Two groups of dairy cows were selected to obtain low-SCC (<200,000 cells/mL) and high SCC (>700,000 cells/mL) milks, which were used to manufacture 2 vats of cheese. The pasteurized milk was evaluated according to the pH, total solids, fat, total protein, lactose, standard plate count, coliforms at 45 degrees C, and Salmonella spp. The cheese composition was evaluated 2 d after manufacture. Lactic acid bacteria, psychrotrophic bacteria, and yeast and mold counts were carried out after 3, 9, 16, 32, and 51 d of storage. Salmonella spp., Listeria monocytogenes, and coagulase-positive Staphylococcus counts were carried out after 3, 32, and 51 d of storage. A 2 x 5 factorial design with 4 replications was performed. Sensory evaluation of the cheeses from low- and high-SCC milks was carried out for overall acceptance by using a 9-point hedonic scale after 8, 22, 35, 50, and 63 d of storage. The somatic cell levels used did not affect the total protein and salt:moisture contents of the cheeses. The pH and moisture content were higher and the clotting time was longer for cheeses from high-SCC milk. Both cheeses presented the absence of Salmonella spp. and L. monocytogenes, and the coagulase positive Staphylococcus count was below 1 x 10(2) cfu/g throughout the storage time. The lactic acid bacteria count decreased significantly during the storage time for the cheeses from both low- and high-SCC milks, but at a faster rate for the cheese from high-SCC milk. Cheeses from high-SCC milk presented lower psychrotrophic bacteria counts and higher yeast and mold counts than cheeses from low-SCC milk. Cheeses from low-SCC milk showed better overall acceptance by the consumers. The lower overall acceptance of the cheeses from high-SCC milk may be associated with texture and flavor defects, probably caused by the higher proteolysis of these cheeses. PMID- 18420605 TI - Production, purification, and characterization of a potential thermostable galactosidase for milk lactose hydrolysis from Bacillus stearothermophilus. AB - Beta-galactosidase, commonly named lactase, is one of the most important enzymes used in dairy processing; it catalyzes the hydrolysis of lactose to its constituent monosaccharides glucose and galactose. Here, a thermostable beta galactosidase gene bgaB from Bacillus stearothermophilus was cloned and expressed in B. subtilis WB600. The recombinant enzyme was purified by a combination of heat treatment, ammonium sulfate fractionation, ion exchange, and gel filtration chromatography techniques. The purified beta-galactosidase appeared as a single protein band in sodium dodecyl sulfate-PAGE gel with a molecular mass of approximately 70 kDa. Its isoelectric point, determined by polyacryl-amide gel isoelectric focusing, was close to 5.1. The optimum temperature and pH for this beta-galactosidase activity were 70 degrees C and pH 7.0, respectively. Kinetics of thermal inactivation and half-life times for this thermostable enzyme at 65 and 70 degrees C were 50 and 9 h, respectively, and the K(m) and V(max) values were 2.96 mM and 6.62 micromol/min per mg. Metal cations and EDTA could not activate this thermostable enzyme, and some divalent metal ions, namely, Fe(2+), Zn(2+), Cu(2+), Pb(2+), and Sn(2+), inhibited its activity. Thiol reagents had no effect on the enzyme activity, and sulfhydryl group blocking reagents inactivated the enzyme. This enzyme possessed a high level of transgalactosylation activity in hydrolysis of lactose in milk. The results suggest that this recombinant thermostable enzyme may be suitable for both the hydrolysis of lactose and the production of galactooligosaccharides in milk processing. PMID- 18420606 TI - Enhancing the lethal effect of high-intensity pulsed electric field in milk by antimicrobial compounds as combined hurdles. AB - High-intensity pulsed electric field (HIPEF) is a nonthermal treatment studied for its wide antimicrobial spectrum on liquid food, including milk and dairy products. Moreover, the antimicrobial effect of HIPEF may be enhanced by combining HIPEF with other treatments as hurdles. Nisin and lysozyme are natural antimicrobial compounds that could be used in combination with HIPEF. Therefore, the purpose of this study was to determine the effect of combining HIPEF with the addition of nisin and lysozyme to milk inoculated with Staphylococcus aureus with regard to different process variables. The individual addition of nisin and lysozyme did not produce any reduction in cell population within the proposed range of concentrations, whereas their combination resulted in a pH-dependent microbial death of Staph. aureus. The addition of nisin and lysozyme to milk combined with HIPEF treatment resulted in a synergistic effect. Applying a 1,200 micros HIPEF treatment time to milk at pH 6.8 containing 1 IU/mL of nisin and 300 IU/mL of lysozyme resulted in a reduction of more than 6.2 log units of Staph. aureus. Final counts resulting from the addition of nisin and lysozyme and applying HIPEF strongly depended on both the sequence of application and the milk pH. Thus, more research is needed to elucidate the mode of action of synergism as well as the role of different process variables, although the use of HIPEF in combination with antimicrobial compounds such as nisin and lysozyme is shown to be potentially useful in processing milk and dairy products. PMID- 18420607 TI - The effect of pasteurization temperature on consumer acceptability, sensory characteristics, volatile compound composition, and shelf-life of fluid milk. AB - The relationship among consumer acceptability, descriptive sensory attributes, and shelf-life was determined for 2% milk pasteurized at 77, 79, 82, and 85 degrees C. Sensory descriptive attributes and volatile compound composition were monitored over the shelf-life of the products to determine if treatments could be differentiated at various times through out the shelf-life of the product. Consumers preferred 79 degrees C milk over other treatments on d 0; however, at d 6 postpasteurization, 79 and 82 degrees C milks were preferred over the 77 degrees C treatment. Consumers were grouped into 8 clusters based on product liking for both d 0 and d 6 evaluations. The largest cluster liked all pasteurization treatments, and 79 degrees C milk was highly acceptable to all consumers who liked milk. Similar sensory descriptors indicated the end of shelf life for all pasteurization treatments even though treatments could be differentiated by descriptors on d 0. This research reveals that altering the pasteurization temperature from 79 degrees C may cause a decrease in consumer acceptability to some consumers. Also, altering pasteurization temperature did not affect shelf-life or sensory descriptors and volatile compound composition at the end of shelf-life. PMID- 18420608 TI - Effect of sex-sorted sperm dosage on conception rates in Holstein heifers and lactating cows. AB - Ejaculates were collected by artificial vagina from 3 Holstein sires and sorted to 90% purity for X-chromosome-bearing spermatozoa (range 88 to 93%) using flow cytometry. Sorted sperm were diluted to 2.1, 3.5, or 5.0 x 10(6) sperm per dose in an egg yolk (20%), Tris, glycerol (7%) extender. Collections were repeated until >600 straws per sperm dose per sire were obtained. Each sperm dose was loaded into color-coded 0.25-mL French straws, with alternate colors used to define treatments across sires. Within sires, straws were packaged at 9 per cane (3 of each color) and strategically allocated to 75 Holstein herds with targets for 50% use in heifers and 50% in lactating cows. Straw color was recorded in the on-farm record-keeping system at the time of insemination. Data were analyzed separately for cows and heifers. Among heifers, a total of 2,125 usable records were retrieved from 51 herds (238 +/- 5.5 services/ sperm dose per sire, range: 218 to 263). Conception rates in heifers were influenced by the sire x sperm dosage interaction. Within sire A, conception rates of heifers were greater for the 5 x 10(6) (59.5%) than for the 2.1 x 10(6) (46.4%) sperm dose and intermediate for the 3.5 x 10(6) sperm dose (52.2%). However, across sires, sperm dosage had no effect on heifer conception rates (46.7, 51.2, and 52.5% for the 2.1, 3.5, and 5.0 x 10(6) sperm dosages, respectively). Among cows, a total of 2,369 services were retrieved from 56 herds (263 +/- 8.8 services/sperm dose per sire, range: 233 to 303). Conception rates of cows (29.4%) were not affected by sire or sperm dosage (27.0, 29.1, and 30.3% for the 2.1, 3.5, and 5.0 x 10(6) sperm dosages, respectively). In conclusion, these data indicate that an increased sperm dosage may enhance virgin heifer conception rates for some (but not all) sires, whereas neither sire nor sexed-sperm dosage affected conception rates of lactating cows. Additional studies of sexed-sperm dosage across a larger sampling of bulls are warranted to determine whether and how such a practice can be implemented cost effectively for the benefit of the dairy industry. PMID- 18420609 TI - Quality of embryos produced from dairy cows fed whole flaxseed and the success of embryo transfer. AB - The objective of the experiment was to determine the effects of fat supplementation on embryo quality of dairy cows and the subsequent success of embryo transfer into recipient heifers fed the same sources of fat. A total of 30 lactating Holstein cows were allotted on d 18 postpartum to 2 groups of 15 donor cows blocked for similar calving dates. Total mixed diets based on silage and fat supplements were fed for ad libitum intake. On a dry matter basis, diets fed to donor cows contained 7.9% whole flaxseed or 2.8% calcium salts of palm oil and those fed to recipient heifers contained 11.4% whole flaxseed or 4.2% calcium salts of palm oil. The experiment with donor cows was carried out between d 18 and 109 of lactation. The experimental diets were fed to 121 recipient heifers from wk 8 before estrus synchronization and superovulation to d 50 of gestation. Dietary fat fed to donor cows had no effect on the number of viable embryos per cow (3.7 +/- 0.5), the number of degenerated embryos per cow (1.8 +/- 0.4), or the number of unfertilized oocytes per cow (2.1 +/- 0.8). But feeding flaxseed decreased fertilization rate (64.3 vs. 78.4%) and the percentage of grade 1 to 2 embryos (56.5 vs. 74.1%) and increased the embryo degeneration percentage (27.4 vs. 18.2%) compared with feeding calcium salts of palm oil. There was no effect of diets fed to donor cows and those fed to recipient heifers for pregnancy rate of heifers. Supplementation with a rich source of n-3 fatty acids decreased quality of embryos from donor lactating dairy cows compared with feeding calcium salts of palm oil, but had no effect on the subsequent pregnancy rate of heifers receiving frozen grade-1 embryos. PMID- 18420610 TI - Continuous lactation in dairy cows: effect on milk production and mammary nutrient supply and extraction. AB - Reports over the past decade have indicated that normal lactational performance can be achieved in genetically superior and high-producing dairy cows, even when the dry period between 2 lactations is omitted. The hypothesis tested in this experiment was that normal lactogenesis I and metabolic function may be achievable in continuously milked high-yielding dairy cows as a result of the genetic selection for lactation performance and hence longevity of mammary epithelial cells. The milk production and mammary nutrient uptake in response to omission of the dry period for cows with an expected peak milk yield higher than 45 kg/d were studied in 28 Holstein dairy cows managed without bovine somatotropin. Performance and metabolic parameters were followed in late gestation and in the following early lactation. Fourteen cows were milked continuously throughout late gestation, and another 14 dairy cows experienced a 7 wk prepartum dry period. Continuous milking during the prepartum period reduced milk production in the following early lactation period by >20%. The reduced milk production could not be readily ascribed to inefficiency of the mechanisms responsible for nutrient uptake by the lactating mammary epithelial cells, nor to systemic endocrine changes. This suggests that lowered mammary nutrient uptake must have been associated with reduced mammary blood flow, metabolic activity, or both, most likely as a result of disturbed lactogenesis I prepartum or lactogenesis II postpartum triggered by as yet unknown local mechanisms. Milk protein content was elevated by 0.4 percentage units in the continuously milked cows. The underlying reason is unknown, but given the current pricing system for milk, it deserves to be further investigated. PMID- 18420612 TI - Variation in plasma concentrations of insulin-like growth factor-I in pasture-fed Holstein cows. AB - The objective of this 5-wk study was to determine dietary effects on plasma concentrations of insulin-like growth factor-I (IGF-I), as well as milk production and milk components in pasture-fed dairy cows. Thirty-two Holstein cows 4 to 5 wk postpartum were randomly assigned to 4 dietary subgroups. Feed was provided twice daily ad libitum at 0900 and 1600 h composed of fresh-cut pasture, meadow hay, and pelleted cereal grain to achieve differing levels of DMI and ME density (LL: 16.6 kg of DMI and 174 MJ of ME; HL: 17.3 kg of DMI and 181.1 MJ of ME; LH: 15.4 kg of DMI and 183.1 MJ of ME; HH: 17.9 kg of DMI and 213.3 MJ of ME, with the first letter indicating DMI and the second ME, and with H indicating high and L indicating low, respectively). The first day cows were placed on their diets was designated d 0. Concentrations of IGF-I were measured in frozen-thawed samples of plasma using a verified ELISA. Dietary treatment had affected plasma concentrations of IGF-I by d 7 with cows on high ME diets having greater IGF-I concentrations at d 14 (83.7 vs. 45.6 ng/mL) than cows on the low ME diets. The level of DMI had less effect on plasma concentrations of IGF-I at d 14 (72.2 vs. 57.1 ng/mL). Dietary treatment effects on these concentrations had stabilized by d 21. Day-to-day variation in mean plasma concentrations of IGF-I within each dietary treatment was low during an intensive period of daily sampling for 14 d (from d 22 to 35). Within-cow day-to-day variation was also low compared with that among cows within the same dietary group and was associated with a high repeatability in the day-to-day concentration of IGF-I in individual cows. Intraclass correlation coefficients for IGF-I ranged from 0.56 (+/- 0.14) to 0.88 (+/- 0.06) with a combined (pooled) value for the 4 subgroups of 0.77 (+/- 0.05). The ME and DMI effects (H vs. L) at d 35 were 79.3 vs. 41.4 and 62.0 vs. 55.7 ng/mL, respectively. Although the ME and DMI differences also affected milk yield and compositional parameters, the effects were not as proportionately great as those measured for IGF-I. Altering the ME or DMI components of the pasture-based diets produced changes in plasma IGF-I concentrations that did not become stabilized for 3 wk, but were then highly repeatable for individual cows within each dietary group. Both observations have relevance to interpreting data related to plasma concentrations of IGF-I in lactating Holstein cows. PMID- 18420611 TI - Growth hormone receptor, insulin-like growth factor (IGF)-1, and IGF-binding protein-2 expression in the reproductive tissues of early postpartum dairy cows. AB - The growth hormone/insulin-like growth factor (IGF) system plays a critical endocrine role controlling nutrient metabolism in dairy cattle. In liver, growth hormone receptor (GHR) and IGF-1 are dynamically regulated by lactation and energy balance. Less is known about the regulation of GHR, IGF-1, and IGF-binding protein mRNA in reproductive tissues (uterus, ovarian follicle, and corpus luteum). The objective was to determine expression patterns for GHR, IGF-1, and IGF-binding protein (IGFBP)-2 mRNA in the liver, uterus, dominant follicle, and corpus luteum in Holstein cows (n = 21) sampled at 3 times during early lactation. The first postpartum ovulation was induced with an injection of GnRH within 15 d of calving. Nine days after ovulation [23 +/- 1 d postpartum; 20 d in milk (DIM)], the liver, uterus, dominant follicle, and corpus luteum were biopsied. Prostaglandin F(2alpha) and GnRH were injected 7 and 9 d after each biopsy to synchronize the second (41 +/- 1 d postpartum; 40 DIM) and third (60 +/ 1 d postpartum; 60 DIM) tissue collections. Total RNA was isolated and used for mRNA analysis by real-time quantitative reverse transcription PCR. Liver had more GHR, IGF-1, and IGFBP-2 mRNA than the reproductive tissues that were tested. Gene expression for GHR, IGF-1, and IGFPB-2 within tissues did not change across the sampling interval (20 to 60 DIM). The only detected change in gene expression across days was for cyclophilin in uterus (increased after 20 DIM). Parity had an effect on gene expression for GHR in corpus luteum. Neither level of milk production nor body condition score affected the amount of GHR, IGF-1, or IGFBP-2 mRNA in the respective tissues. The repeatability of gene expression within a tissue was 0.25 to 0.5 for most genes. In most instances, expression of a single gene within a tissue was correlated with other genes in the same tissue but was not correlated with the same gene in a different tissue. We did not find evidence for major changes in gene expression within reproductive tissues in postpartum cows. Differences between cows (independent of their BCS and milk production) accounted for a major portion of the variation that we observed. PMID- 18420613 TI - Effect of restricted feeding and monopropylene glycol postpartum on metabolic hormones and postpartum anestrus in grazing dairy heifers. AB - This study was designed to determine the effects of feed restriction and monopropylene glycol (MPG) supplementation on the reproductive, milk production, and somatotropic axes in dairy heifers postpartum. At calving, 49 Holstein Friesian heifers were allowed either unrestricted (UNR; n = 18) or restricted access to pasture with (RES+MPG; n = 13) or without (RES; n = 18) MPG supplementation (250 mL drenched twice daily for 150 d). The average body condition score (BCS) of the heifers was 5.3 +/- 0.2 on a scale from 1 to 10 (where 1 = emaciated and 10 = obese). Body condition score and body weight were similar among the groups at calving and decreased after calving for all groups. However, body weight loss was around 10% greater for the RES and RES+MPG groups from wk 3 to 12 compared with UNR group. The length of the postpartum anestrous interval was similar for all groups (47, 51, and 45 +/- 5 d for the UNR, RES, and RES+MPG, respectively). Average milk production, protein, fat, and lactose yields during the first 12 wk postpartum were greater in the UNR group than in the RES and RES+MPG groups. Feed restriction affected plasma concentrations of insulin, with lower concentrations in the RES group compared with the UNR group. There were no differences in plasma concentrations of insulin between the RES+MPG group and the UNR or RES groups. An effect of feed restriction was observed on insulin like growth factor-I concentrations and also a treatment by time interaction with a changing pattern through time as concentrations in the UNR group increased relative to the RES and RES+MPG groups. There were no differences in growth hormone concentrations among the groups. Glucose concentrations were lower in the RES group when compared with RES+MPG and UNR groups and this difference lessened over time. Plasma concentrations of nonesterified fatty acids were greater in the RES group compared with the RES+MPG and UNR groups. Leptin concentrations in the UNR group were greater than in the RES and RES+MPG groups. Hepatic growth hormone receptor 1A, total growth hormone receptor, and insulin-like growth factor-I relative mRNA expressions decreased postpartum with no effect of feed restriction, MPG supplementation, or interaction between time and treatment. During a challenge with MPG, insulin secretion was stimulated but no effect on postpartum anestrous interval in the treatment groups was observed. It was concluded that restricted pasture availability postpartum in dairy heifers calving in optimal BCS had no effect on the postpartum anestrous interval. It did however decrease milk production; thus, we can infer that monopropylene glycol supplementation does not act to prevent loss of milk yield. PMID- 18420614 TI - Evaluation of serotonin as a feedback inhibitor of lactation in the bovine. AB - Serotonin (5-HT), a neurotransmitter synthesized from tryptophan, has been proposed as a feedback inhibitor of lactation. We determined that the gene coding for tryptophan hydroxylase 1, the rate-limiting enzyme for 5-HT synthesis, is expressed in bovine mammary epithelial cells in vitro and is upregulated by prolactin. In addition, 5-HT reduced the expression of alpha-lactalbu-min and casein genes in vitro. Furthermore, inhibiting 5-HT synthesis with p chlorophenylalanine or blocking the 5-HT receptor with methysergide (METH) increased milk protein gene expression. We then evaluated effects of intramammary 5-HT or METH infusion on production and milk composition in 6 multiparous Holstein cows. Cows were assigned to a repeated measures design of contralateral intramammary infusions of METH (20 mg/quarter per d) or saline for 3 d followed by a 7-d washout period before administering 5-HT (50 mg/quarter/d) or SAL for 3 d. For each udder half, milk yield was recorded twice and composition was determined once per day. Blood samples were harvested each day for plasma to determine glucose and nonesterified fatty acid concentrations. Evaporative heat loss, respiration rate, left and right udder temperatures, and rectal temperatures were obtained after each milking to evaluate possible systemic effects of infusions. During METH and saline infusions milk yield increased 10.9%. During 5-HT and saline infusion milk yield decreased 11.1%. Milk yield and physiological responses suggested intramammary 5-HT and METH doses were high enough to cause systemic effects. Infusing saline, METH, and 5-HT increased milk SCC. Infusing 5-HT tended to reduce mean lactose concentration (4.3 vs. 4.6%) relative to saline. Milk protein content was decreased by METH and SAL (2.0%) and was increased (5.8%) by 5-HT followed by a 33% decrease postinfusion. Infusion of METH increased evaporative heat loss 11%, which decreased 11% postinfusion. Infusions of 5-HT or METH did not affect plasma nonesterified fatty acid or glucose concentrations, respiration rate, or milk fat content. We conclude 5-HT infusion reduced milk synthesis, whereas blocking the 5-HT receptor with METH increased milk synthesis. Doses of 5-HT and METH used in this study likely resulted in systemic effects. These data support the concept that 5-HT is a feedback inhibitor of lactation in the bovine. PMID- 18420615 TI - Production and health of cows given monensin prepartum and a high-energy diet postpartum. AB - The object of this study was to evaluate the impact of monensin administration on the early lactation performance of cows maintained on a high-energy diet, and on health traits during the transition period. Cows (n = 168; parity 3.3 +/- 1.4, initial body condition score 3.1 +/- 0.08, and milk yield of 34.3 kg/d +/- 0.9 for multiparous cows in the preceding lactation) were divided into control and monensin treatment groups. A controlled-release capsule supplying 335 mg of monensin/d for 95 d was inserted into the rumen of monensin-treated cows 30 d before the expected calving. Blood samples were obtained 2 h after feeding on d 14 prepartum and on d 7, 14, and 50 postpartum. Plasma glucose concentration was 3% higher (58 +/- 0.5 vs. 56.4 +/- 0.5 mg/dL) and beta-hydroxybutyric acid was 17% (6.7 +/- 0.3 vs. 8.0 +/- 0.3 mg/dL) lower in monensin-treated than control cows. Plasma glucose was 10% higher (60.0 +/- 0.6 vs. 54.5 +/- 0.3 mg/dL) and beta-hydroxybutyric acid was 16% lower (6.8 +/- 0.3 vs. 7.9 +/- 0.2 mg/dL) in primiparous than multiparous cows. Plasma nonesterified fatty acid concentration (measured only in primiparous cows) was 17% lower (287 +/- 15 vs. 336 +/- 17 muEq/L) in treated than in control cows. Rate of ketosis incidence was 60% lower (8 vs. 21%) in monensin-treated than in control cows, and the proportion of control cows that required a supply of glucogenic precursors was 3-fold higher than in monensin-treated cows. The body condition score was 3.1 +/- 0.05, 2.7 +/- 0.05, and 2.4 +/- 0.05 on d 60 prepartum and d 7 and 50 postpartum, respectively, and was not affected by treatment. During the first 5 mo of lactation, milk yield was 7% higher (37.6 +/- 0.6 vs. 35.2 +/- 0.6 kg/d) in monensin-treated cows than in control cows. Our results showed that monensin administration, as a controlled release capsule in prepartum cows, can be beneficial, even if these cows are maintained on a high-energy diet during the subsequent lactation. PMID- 18420616 TI - Activation of immune cells in bovine mammary gland secretions by zymosan-treated bovine serum. AB - Mastitis, caused by bacterial infection of the mammary gland, is a major disease of dairy cattle. The greatest risks of intramammary infection occur at the end of lactation and at the initiation of the next lactation when the cow calves. Treating serum with zymosan (yeast cell wall preparation) causes the complement to cleave, allowing this serum to serve as a source of complement fragment 5a (C5a), a potent chemoattractant and activator of the immune system. Our hypothesis was that intramammary infusion of zymosan-treated serum (ZTS) would recruit polymorphonuclear neutrophils (PMN) and generate prolonged activity in lymphocytes within the mammary gland. Ultimately this could help prevent bacterial infections in cows at dry-off and at the initiation of lactation. Two ipsilateral quarters of the mammary gland of each cow were infused with ZTS (12.5 mL/quarter), and 2 contralateral quarters were infused with saline in 8 cows shortly after lactation ended. Mammary secretions were collected periodically throughout the dry period and the first 2 wk of the next lactation. Activation status of lymphocytes and PMN in those secretions was assessed based on the intracellular presence or absence of IFN-gamma and IL-8 as determined by flow cytometry. The ZTS infusion greatly increased PMN numbers in mammary secretions for the first week only. The percentage of IFN-gamma positive lymphocytes and PMN, and the percentage of IL-8 positive PMN, exhibited a sustained increase in secretions from ZTS-treated quarters through the first 2 wk of lactation. The ZTS can stimulate PMN and lymphocyte-mediated immune defense mechanisms in the mammary gland, which may provide a useful means of preventing new intramammary infections during the dry period as well as at the initiation of lactation. PMID- 18420617 TI - Relationship between milk lactoferrin and etiological agent in the mastitic bovine mammary gland. AB - Bovine mastitis is one of the most deleterious diseases for dairy herds and is mainly caused by contagious and environmental bacterial pathogens. Among contagious bacteria, Staphylococcus aureus is the most prevalent, whereas the main environmental mastitis pathogens are Streptococcus uberis and Escherichia coli. Bovine lactoferrin (bLF) is an approximately 80-kDa glycoprotein present in milk that participates in the innate response of the mammary gland against bacterial infection. The objectives of the current study were to analyze potential changes in bLF milk concentration, which would constitute a response of the mammary gland toward mastitis induced by different etiologic agents, and to evaluate a possible relation between this response and pathogen susceptibility to bLF. Microbiology analysis and bLF quantification in milk from different bovine mammary gland quarters were performed. Infected quarters presented greater concentrations of bLF compared with those from microbiologically negative quarters. Analysis of individual pathogen contributions showed that most of this increase was attributable to Strep. uberis intra-mammary infection. The ability of mammary gland cells to synthesize bLF in response to Strep. uberis challenge was demonstrated by immunodetection of the protein in in vitro infection experiments. Susceptibility of Strep. uberis, E. coli, and Staph. aureus to the antimicrobial activity of bLF was determined by growth inhibition assays conducted with 4 different isolates of each species. Whereas Staph. aureus and E. coli were shown to be susceptible to this protein, Strep. uberis appeared to be resistant to the antimicrobial activity of bLF. Molecular typing of the 4 Strep. uberis isolates used throughout this study showed that this result was representative of the species and not exclusive of a particular strain. Results presented herein suggest that different bacteria species may elicit different mammary gland responses mediated by bLF secretion and that Strep. uberis has probably adapted to this immune reaction by developing resistance to bLF inhibitory action. PMID- 18420618 TI - Effect of different flooring systems on weight and pressure distribution on claws of dairy cows. AB - Weight and pressure distribution on the claw were studied in Swedish Holsteins housed in different flooring systems. A total of 127 cows housed in different sections of the experimental barn were used. Each section had different flooring in the walking and standing areas. There were rubber mats or abrasive mastic asphalt flooring on the alleys or a low-abrasive slatted concrete floor. Some sections had feed-stalls equipped with rubber mats; other sections did not. The vertical ground reaction force, contact area, and average contact pressure were determined on the left hind foot using the I-Scan system and analyzed with the F scan system. These determinations were made in each of the following 3 zones of the claw: bulb, wall, and sole. Most of the weight on claws exposed to concrete floors was carried by the bulb (37.4 +/- 3.5 and 18.3 +/- 2.9% of weight exerted on the foot in the lateral and medial claw, respectively) and the wall zone (20.0 +/- 2.6 and 13.4 +/- 2.4% on lateral and medial claw, respectively). The weight and pressure distribution in cows kept on sections with rubber covered alleys but passing daily over the asphalt floor on their way to the milking parlor did not differ in any zones, except for lateral bulbs, compared with those exposed to slatted concrete alone. Still, the weight bearing of the sole zone in cows kept on rubber mats without access to asphalt was less than that of cows kept on concrete slatted floors (5.1 +/- 0.7 vs. 12.7 +/- 1.1% and 1.1 +/- 0.5 vs. 8.7 +/ 0.7% in lateral and medial claws, respectively). In cows kept on asphalt flooring without feed-stalls, most weight was exerted to the sole zone (36.2 +/- 2.9 and 22.2 +/- 1.8% in lateral and medial claws, respectively). Feed-stalls in combination with asphalt flooring yielded a decreased total contact area (30.1 +/ 1.2 cm(2)) compared with asphalt floors without feed-stalls (35.7 +/- 1.2 cm(2)). The largest total contact area was obtained on the asphalt floor without feed-stalls, resulting in a lower contact pressure (39.8 +/- 2.3 N/cm(2)) than in claws exposed to concrete (66.0 +/- 2.7 N/ cm(2)) or rubber mats (56.7 +/- 1.7 N/cm(2)). In conclusion, housing with abrasive floors resulted in claws with increased contact area at the sole surface and therefore, decreased contact pressure, but reduced the weight-bearing role of the strongest part of the claw capsule, the claw wall. PMID- 18420619 TI - Evaluation of free-stall mattress bedding treatments to reduce mastitis bacterial growth. AB - Bacterial counts were compared in free-stall mattresses and teat ends exposed to 5 treatments in a factorial study design on 1 dairy farm. Mattresses in five 30 cow groups were subjected to 1 of 5 bedding treatments every other day: 0.5 kg of hydrated limestone, 120 mL of commercial acidic conditioner, 1 kg of coal fly ash, 1 kg of kiln-dried wood shavings, and control (no bedding). Counts of coliforms, Klebsiella spp., Escherichia coli, and Streptococcus spp. were lowest on mattresses bedded with lime. Mattresses bedded with the commercial acidic conditioner had the next lowest counts for coliforms, Klebsiella spp., and Streptococcus spp. Wood shavings and the no-bedding control had the highest counts for coliform and Klebsiella spp. Compared with wood shavings or control, fly ash reduced the counts of coliforms, whereas for the other 3 bacterial groups, the reduction was not always significant. Streptococcus spp. counts were greatest in the control group and did not differ among the shavings and fly ash groups. Teat swab results indicated that hydrated lime was the only bedding treatment that significantly decreased the counts of both coliforms and Klebsiella spp. There were no differences in Streptococcus spp. numbers on the teats between any of the bedding treatments. Bacterial populations grew steadily on mattresses and were generally higher at 36 to 48 h than at 12 to 24 h, whereas bacterial populations on teats grew rapidly by 12 h and then remained constant. Hydrated lime was the only treatment that significantly reduced bacterial counts on both mattresses and teat ends, but it caused some skin irritation. PMID- 18420620 TI - A longitudinal field study to evaluate the diagnostic properties of a quantitative real-time polymerase chain reaction-based assay to detect Staphylococcus aureus in milk. AB - Bacteriological culture as a diagnostic tool for chronic infections with Staphylococcus aureus intramammary infection is not completely satisfactory. The cyclical shedding pattern of Staph. aureus with intervals of low excretion is considered to be the main reason. We recently developed a novel assay for Staph. aureus in milk, based on real-time quantitative PCR (QPCR). In a longitudinal study of chronically infected cows we evaluated the diagnostic properties of this test under field conditions. Diagnostic sensitivity of the novel test proved to be very high with a value of 99.4%; diagnostic specificity was 97.1%. In addition, the shedding patterns of Staph. aureus for the sampling period were analyzed. Using log(10)-transformed QPCR data and plotting them across sampling time revealed a sinusoidal shedding pattern in 6 of 11 naturally infected quarters. Shedding patterns obtained by QPCR and by bacteriological culture were synchronous. In conclusion, the novel test has a very high diagnostic sensitivity and specificity so that quarters chronically infected with Staph. aureus are reliably detected, independent from their actual shedding quantity. PMID- 18420621 TI - Short communication: effect of stocking density on indices of cow comfort. AB - The objective of this study was to evaluate the effects of increasing stocking density on indices of cow comfort measured over a 24-h period, during peak lying time (0000 to 0400 h), and 1 h after the afternoon milking. Holstein cows (n = 136) were assigned to 1 of 4 pens, and stocking densities of 100, 113, 131, and 142% were applied in a 4 x 4 Latin square. Video data were recorded continuously for 2 d after 5 d of acclimation to stocking density and analyzed with 10-min scan samples for the percentage of cows lying in a stall, standing in a stall, standing in the alley, and eating at the manger. Percentage of cows standing idly in the alley increased as stocking density increased above 113% (10.9, 12.0, 14.4, and 16.5% for 100, 113, 131, and 142% stocking density, respectively). Cow comfort index (CCI; cows lying down/cows contacting stalls) and stall standing index (SSI; cows standing in stall/cows contacting stalls) differed little as stocking density increased, but stall use index (SUI; cows lying in stall/cows in pen not eating) decreased beyond 113% stocking density (70.1, 70.2, 68.6, and 66.3 for 100, 113, 131, and 142% stocking density, respectively). During peak lying time, SUI decreased with increasing (80.3, 79.5, 74.8, and 69.6 for 100, 113, 131, and 142% stocking density, respectively) stocking density above 113%, whereas CCI and SSI showed little response. None of the indices varied by stocking density when they were assessed at 1 h after milking. These results suggest that more than 1 index of cow comfort may be needed at higher stocking densities to assess both stall usage and cows standing idly in an alley. At higher stocking densities, SUI was reduced, because it reflected not only stall usage but the number of cows standing idly in an alley and not actively feeding and unable to access a stall. The CCI and SSI appear to assess actual stall usage (% cows lying or standing) across the range of stocking densities evaluated in this study. PMID- 18420622 TI - Technical note: immunomagnetic procedure for positive selection of macrophages in ovine milk. AB - A simple immunomagnetic procedure was developed to select macrophages from ovine milk by using a non-specific magnetic positive separation technique. Samples of ewe bulk milk were collected during early, mid, and late lactation; milk samples were centrifuged at 2,000 x g for 30 min at 4 degrees C; the fatty fraction and supernatant were removed, and each pellet was dissolved in 500 microL of pH 7.4 phosphate buffered saline + 0.02% NaN(3). Cells were targeted for selection by using mouse-IgG anti-ovine macrophages. Several trials, testing 2 different fluorochrome-conjugated antibodies [i.e., mouse anti-human CD14:R-Phycoerythrin (RPE) (MCA1568PE, Serotec) and F(ab')2 rabbit anti-mouse IgG:RPE (STAR12A, Serotec)] and 3 different labeling procedures, were performed to evaluate the purity of samples by flow cytometry. A morphological test was carried out by direct microscopic count in enriched fraction smears stained with May-Grunwald Giemsa stain to confirm the presence of macrophages. The method described in the present technical note can be considered an innovative application to obtain a single-cell population of high purity selected from all the somatic cells in milk. PMID- 18420623 TI - Effects of feeding prepubertal heifers a high-energy diet for three, six, or twelve weeks on feed intake, body growth, and fat deposition. AB - The objective was to determine the effects of feeding prepubertal dairy heifers a high-energy diet for a duration of 0, 3, 6, or 12 wk on feed intake, growth, and fat deposition. We also used feed composition, daily intake, and body growth data to evaluate the nutritional model of the 2001 National Research Council (NRC) Nutrient Requirements of Dairy Cattle. Holstein heifers (age = 11 wk; body weight = 107 +/- 1 kg) were assigned to 1 of 4 treatments (n = 16/treatment) designated H0, H3, H6, and H12 and fed a low-energy diet for 12, 9, 6, or 0 wk, followed by a high-energy diet for 0, 3, 6, or 12 wk, respectively. Four heifers were killed initially (11 wk of age) and 64 heifers were killed at the end of the treatment period (23 wk of age). The low-energy diet was formulated to achieve 0.6 kg of average daily gain and contained 16% crude protein, and 45% neutral detergent fiber. The high-energy diet was formulated to achieve an average daily gain of 1.2 kg and contained 18% crude protein and 23% neutral detergent fiber. Actual daily gains averaged over the 12-wk treatment period were 0.64, 0.65, 0.83, and 1.09 kg for the H0, H3, H6, and H12 groups, respectively. Body weight, withers height, hip width, carcass weight, liver weight, and perirenal fat increased in heifers fed a high-energy diet for a longer duration. In addition, percentage of fat increased and percentage of protein decreased in rib sections with a longer duration on the high-energy diet. Uterine and ovarian weights adjusted for body weight decreased when heifers were fed the high-energy diet for a longer duration. The 2001 NRC underestimated dry matter intake of the high-energy diet and overestimated dry matter intake of the low-energy diet. On the basis of actual intakes of each diet, the NRC slightly underestimated gain for the low energy diet and overestimated gain by 40% for the high-energy diet. The likely explanation for this is that the NRC underestimated the proportion of gain that was fat in the heifers fed the high-energy diet and therefore predicted more body gain per unit of energy intake. We concluded that feeding a high-energy diet for a short duration altered body growth and fat deposition in a time-dependent, linear manner consistent with feeding a high-energy diet for a long duration. PMID- 18420624 TI - Effects of feeding prepubertal heifers a high-energy diet for three, six, or twelve weeks on mammary growth and composition. AB - The experimental objective was to determine the effects of feeding prepubertal dairy heifers a high-energy diet for 3, 6, or 12 wk on mammary growth and composition. Holstein heifers (age = 11 wk; body weight = 107 +/- 1 kg) were assigned to 1 of 4 treatments (n = 16/ treatment). The treatment period lasted 12 wk and treatments were H0 (low-energy diet fed for 12 wk, with no weeks on the high-energy diet); H3 (low-energy diet fed for 9 wk, followed by the high-energy diet for 3 wk); H6 (low-energy diet fed for 6 wk, followed by the high-energy diet for 6 wk); and H12 (high-energy diet for all 12 wk). The low- and high energy diets were formulated to achieve 0.6 and 1.2 kg of average daily gain, respectively. Heifers were slaughtered at 23 wk of age and mammary tissue was collected. A longer duration of feeding the high-energy diet increased total mass of the mammary gland, extraparenchymal fat, and intraparenchymal fat, but did not alter the mass of fat-free parenchymal tissue. When adjusted for carcass weight to reflect differences in physical maturity, the mass of fat-free parenchymal tissue decreased in a linear fashion with a longer duration on the high-energy diet. Total masses of mammary parenchymal DNA and RNA were not different. However, after adjustment for carcass weight, the masses of DNA and RNA decreased as heifers were fed the high-energy diet for a longer duration. The percentages of epithelium, stroma, and lumen, the number of epithelial structures, and the developmental scores of mammary parenchymal tissue were not different among treatments. However, the percentage of proliferating epithelial cells in the terminal ductal units, as indicated by Ki-67 labeling, decreased as heifers were fed the high-energy diet for a longer duration. We concluded that feeding prepubertal heifers a high-energy diet for a longer duration resulted in a linear decrease in both the percentage of mammary epithelial cells that were proliferating and in the mass of fat-free mammary parenchyma per unit of carcass. High-energy feeding hastens puberty and, in this study, decreased mammary epithelial cell proliferation in areas of active ductal expansion. These data are consistent with the idea that feeding heifers a high-energy diet will reduce mammary parenchymal mass at puberty. PMID- 18420625 TI - Effect of feeding corn, hull-less or hulled barley on fermentation by mixed cultures of ruminal microorganisms. AB - Increased demands for corn grain warrant the evaluation of alternative grain types for ruminant production systems. This study was conducted to determine the effects of hulled and hull-less barley (Hordeum vulgare L.) cultivars compared with corn (Zea mays L.) as an alternative grain type on fermentation in cultures of mixed ruminal microorganisms. Three continuous fermentors were fed 14 g of dry feed per day (divided equally between 2 feedings) consisting of alfalfa (Medicago sativa L.) hay pellets (40% of dry matter) and 1) ground corn, 2) hulled barley, or 3) hull-less barley concentrate (60% of dry matter) in each fermentor. Following an adaptation period of 5 d, culture samples were taken at 2 h after the morning feeding on d 6, 7, and 8 of each period for analysis. A second run of the fermentors followed the same treatment sequence to provide replication. Culture pH was reduced with corn (5.55) and did not differ between barley cultivars (average pH 5.89). Total volatile fatty acid concentration and acetate to propionate ratio were not different across grain type or barley cultivar with the exception of greater total volatile fatty acid concentrations with hull-less barley. Corn produced less methane (14.6 mmol/d) and ammonia-N (7.3 mg/100 mL) compared with barley (33.1 mmol/d and 22 mg/100 mL, respectively); methane was greater with hull-less barley but ammonia-N concentration was similar between the 2 barley cultivars. Hull-less barley had greater digestibility compared with hulled barley, and corn had reduced digestibility compared with barley. Concentrations of C18:0 were greater and those of C18:1 and C18:2 lesser in cultures fed hulled and hull-less barley compared with corn. Our data indicate that grain type and barley cultivar have an impact on ruminal fermentation. The lesser starch concentration of barley minimized the drop in culture pH and improved digestibility. PMID- 18420626 TI - Effects of the chop lengths of alfalfa silage and oat silage on feed intake, milk production, feeding behavior, and rumen fermentation of dairy cows. AB - Effects of chop length (shorter: 6 mm, or longer: 19 mm) of alfalfa silage and oat silage were determined in 16 mid-lactation Holstein cows, 4 of which were rumen cannulated, using a replicated 4 x 4 Latin square design with a 2 x 2 arrangement of treatments. Experimental periods were 21 d long and consisted of 14 d of adaptation and 7 d of sampling. Cows received a total mixed ration containing [dry matter (DM) basis] 42.0% barley grain-based energy supplement, 10% protein supplement, and 24% of DM longer chop or shorter chop alfalfa silage and 24% of DM longer chop or shorter chop oat silage. Rumen pH was measured continuously, and rumen liquid flow rates were determined in rumen-cannulated cows. Feeding behavior was determined by videotaping, and meal patterns were determined by continuously weighing the feed in the bunk of 8 cows. Reducing the chop length of alfalfa silage and oat silage reduced the average geometric particle length from 14.2 to 10.9 mm and from 13.4 to 10.4 mm, respectively. Reducing the alfalfa chop length did not affect feed intake, whereas reducing the oat silage chop length increased DM intake from to 19.4 to 21.2 kg/d. Reducing the chop lengths of alfalfa silage and oat silage chop length did not affect milk production, rumen fermentation, feeding behavior, meal patterns, and blood metabolites. Daily milk yield, milk fat percentage, and milk protein percentage averaged 36.1 kg/d, 3.00%, and 3.16%, respectively, across diets. The low milk fat percentages suggest that the diets induced subacute ruminal acidosis. This was also substantiated by the rumen pH, which was below 5.6 for more than 122 min/d for all diets. The onset of subacute ruminal acidosis despite apparently adequate dietary neutral detergent fiber content and particle size distribution as well as the long duration of chewing might be attributed to sorting against long feed particles. PMID- 18420627 TI - Timothy hay with a low dietary cation-anion difference improves calcium homeostasis in periparturient Holstein cows. AB - The current study was undertaken to evaluate the effects of feeding timothy (Phleum pratense L.) hay differing in dietary cation-anion difference (DCAD) on the capability of cows to maintain calcium homeostasis around parturition. We hypothesized that feeding low-DCAD timothy hay during the prepartum period would induce a mild metabolic acidosis prepartum and improve calcium homeostasis postpartum with no effect on dry matter intake. Forty-one dry pregnant Holstein cows entering their second lactation or greater were used in a randomized complete block design. Timothy hay was obtained from an established timothy stand under a pivot irrigation system. Low-DCAD timothy hay was produced by fertilizing the area between the second and third pivot towers at a rate of 224 kg of CaCl(2)/ha, and control timothy hay (high DCAD) was grown on the area between the fourth and fifth pivot towers of the same field. The chloride concentration was 1.07 and 0.15% on a dry matter (DM) basis, and the DCAD was 1.2 and 21.6 mEq/100 g of DM for the low- and high-DCAD timothy hay, respectively. Experimental diets, containing timothy hay at 63% of dietary DM, were fed ad libitum starting 30 d before the expected calving date. The DCAD values were 1.6 vs. 14.5 mEq/100 g of DM for the low- and high-DCAD timothy-based diets, respectively. At the beginning of the study, urine pH and blood bicarbonate concentration averaged 8.22 +/- 0.06 and 28.5 +/- 0.3 mM, respectively. The low-DCAD timothy diet decreased urine pH compared with the high-DCAD timothy diet on d 21 (7.75 vs. 8.31), d 14 (7.69 vs. 8.22), and d 7 (7.50 vs. 8.19) before calving, and it also decreased the prepartum blood bicarbonate concentration by 2 mM. In addition, cows fed the low DCAD timothy diet had greater blood ionized calcium concentration prepartum (1.22 vs. 1.19 mM), greater blood ionized calcium concentration at 0 and 8 h after calving, and similar prepartum dry matter intake. These results indicate that timothy hay differing in DCAD affects the acid-base balance of periparturient dairy cows, and that low-DCAD timothy hay improves calcium homeostasis postpartum with no negative effect on dry matter intake. PMID- 18420628 TI - Efficacy of liquid feeds varying in concentration and composition of fat, nonprotein nitrogen, and nonfiber carbohydrates for lactating dairy cows. AB - In trial 1, we evaluated the efficacy of a liquid feed (LF) containing cane molasses and corn steep liquor as carriers of suspended white grease (WG) without or with urea (U) or with soybean lipid (SL; a byproduct of soybean processing) compared with roasted soybeans plus tallow blended into respective concentrates in a 16-wk lactation study. The dry matter intake (DMI) and milk production for LF diets were either similar to or greater than respective controls, although SL decreased milk fat percentage. In trial 2, we compared LF without fat to LF plus WG or SL and also evaluated the dose response to increasing amount of LF + WG in a 16-wk lactation trial in which the LF products were added to respective total mixed rations. The DMI was increased and then decreased (quadratic response) with increasing LF + WG without a linear response. However, production of milk, protein, and fat increased linearly with corresponding quadratic responses, which we interpret to be a result of a limiting returns response from DMI and density of net energy for lactation. When LF plus SL was fed, milk fat percentage and yield decreased compared with the comparable amount of LF + WG. In a 12-wk lactation study (trial 3), we added 3.25 or 6.5% of the dry matter as LF (a different but generally similar product than the previous trials and without fat) to diets formulated to maintain comparable ruminal nonstructural carbohydrate digestibility by adding soybean hulls to decrease nonfiber carbohydrates (NFC) concentration; the 6.5% LF diet was without or with Rumensin (11.5 g/909 kg of dry matter). When 3.25% LF was added but NFC was decreased from 40 to 37%, cows increased DMI and production of milk fat. Adding Rumensin decreased DMI but maintained milk fat yield compared with its 6.25% LF control without Rumensin. In trials 1 and 3, apparent total tract nutrient digestibility was not affected by treatment. In conclusion, feeding LF at about 5% (trial 2, which contained WG, 1.6% added sugar) or 3.25% (trial 3, 1.7% added sugar) generally increased DMI and maintained or increased production of milk, protein, and fat. PMID- 18420629 TI - Effect of trace mineral source on lactation performance, claw integrity, and fertility of dairy cattle. AB - Two hundred fifty multiparous and primiparous cows were assigned to a study at approximately 70 d prepartum to determine the effect of trace mineral source on lactation performance, claw integrity, and fertility. Cows received treatments from 3 wk prepartum through wk 35 postpartum. Treatments consisted of 1) all supplemental Zn, Mn, Cu, and Co provided in sulfate form (Sulfate) and 2) 360 mg of Zn, 200 mg of Mn, 125 mg of Cu, and 12 mg of Co supplied daily by Sulfate minerals replaced with similar amounts of minerals supplied by Availa-4 (CTM). Individuals involved with daily animal care or data recording, or both, were blinded to treatment assignments. Cows from all treatments were housed in common pens, and treatments were dispensed to cows via a computerized feeder. All claws of cows were examined before treatment administration and at 16 and 36 wk postpartum by personnel trained in identifying claw lesions. Cows fed the CTM diet tended to produce more milk and energy-corrected milk than cows fed the Sulfate diet. Cows fed the CTM diet also produced more milk protein and solids (fat + protein) than cows fed the Sulfate diet. Replacing Sulfate minerals with those supplied by CTM decreased incidence of sole ulcers at wk 36 postpartum and tended to decrease incidence of interdigital dermatitis at wk 16 and 36 postpartum. Severity of heel erosion tended to be less for cows fed CTM than cows receiving the Sulfate diet. Despite first service conception rates tending to be greater for cows fed the Sulfate diet, there was no effect of treatment on rate of conception. A greater percentage of cows fed the Sulfate diet tended to be culled from the herd before wk 36 postpartum than cows fed the CTM diet. Replacing Sulfate minerals with CTM resulted in improved lactation performance and claw integrity. PMID- 18420630 TI - Extent of methionine limitation in peak-, early-, and mid-lactation dairy cows. AB - Five multiparous, ruminally and duodenally cannulated Holstein cows were assigned to 5 x 5 Latin squares at wk 2 (experiment 1), wk 11 to 13 (experiment 2), and wk 17 to 19 postpartum (experiment 3) to determine extent of Met limitation. Treatments were duodenally infused and consisted of 10 g/d of l-Lys plus 0, 3.5, 7.0, 10.5, or 16.0 g/d of dl-Met in experiments 1 and 2 and 8 g/d of l-Lys plus 0, 5, 10, 15, or 20 g/d of dl-Met in experiment 3. Calculated Lys contributions to total AA (TAA) in duodenal digesta for control treatments were 8.6, 7.5, and 9.0% for experiments 1, 2, and 3, respectively. Methionine contributions to TAA for the 5 infusion treatments were 1.9, 2.1, 2.2, 2.4, and 2.7% for experiment 1; 2.1, 2.3, 2.4, 2.5, and 2.7% for experiment 2; and 1.8, 2.0, 2.2, 2.4, and 2.5% for experiment 3, respectively. Milk protein yield increased linearly in experiments 1 and 2, indicating that Met contribution to TAA in duodenal digesta for maximal milk protein synthesis exceeded 2.7 for early-lactation cows. In experiment 2, a quadratic relationship was found between level of infused Met and milk protein content, with the response reaching a plateau when 12.2 g of Met was infused, corresponding with a Met contribution to TAA in duodenal digesta of 2.4%. In experiment 3, milk protein content increased quadratically, but milk yield declined linearly with increasing levels of infused Met; hence, milk protein yield was unaffected by treatment. The calculated plateau point of the milk protein content response curve was determined to be 12.4 g of infused Met, which corresponds to a Met contribution to TAA in duodenal digesta of 2.3%. Experiment 3 results indicate that the required level of Met in duodenal digesta for maximizing milk protein yield is lower than that required for maximizing milk protein content. PMID- 18420631 TI - Metabolism of dairy cows as affected by prepartum dietary carbohydrate source and supplementation with chromium throughout the periparturient period. AB - Holstein cows (n = 72) entering second or later lactation were used to determine whether metabolic indices and hepatic capacities for oxidation and gluconeogenesis from propionate are affected by source of carbohydrate in the prepartum diet and chromium-l-methionine (Cr-Met) supplementation throughout the periparturient period. Cows were fed prepartum diets as total mixed rations with the concentrate portion based either on starch-based cereals [high nonfiber carbohydrate (NFC); 1.59 Mcal/kg of net energy for lactation (NE(L)), 14.4% crude protein (CP), 40.3% NFC] or nonforage fiber sources (low NFC; 1.54 Mcal/kg of NE(L), 14.5% CP, 33.6% NFC) from 21 d before expected parturition until parturition. After parturition all cows were fed a common lactation total mixed ration (1.74 Mcal/kg of NE(L), 16.5% CP, 40.0% NFC). The Cr-Met was supplemented once daily via gelatin capsule at dosages of 0, 0.03, or 0.06 mg of Cr/kg of BW(0.75). Thus, treatments were in a 2 (carbohydrate source) x 3 (Cr-Met) factorial arrangement. There was no effect of prepartum carbohydrate source on pre- and postpartum plasma concentrations of glucose, nonesterified fatty acids (NEFA), beta-hydroxybutyrate (BHBA), insulin, glucagon, or insulin to glucagon ratio. However, cows fed the low NFC diet during the prepartum period tended to have greater plasma NEFA and lower BHBA concentrations postpartum. Liver glycogen concentrations tended to be greater on d 1 postpartum for cows fed low NFC prepartum. Supplementing 0.03 mg/kg of BW(0.75) of Cr as Cr-Met increased prepartum plasma glucose and glucagon concentrations and tended to decrease prepartum plasma NEFA concentrations compared with either 0 or 0.06 mg of Cr/kg of BW(0.75). Postpartum plasma glucose concentrations decreased linearly and glucagon concentrations were increased quadratically by administering increasing amounts of Cr-Met. Supplementing Cr-Met did not affect prepartum plasma concentrations of insulin or BHBA, postpartum NEFA or BHBA, or liver composition. There was an interaction of prepartum carbohydrate source and Cr-Met supplementation such that in vitro hepatic conversion of [1-(14)C]propionate to both CO(2) and glucose was similar or increased when Cr-Met was supplemented to cows fed the low NFC diet but decreased when Cr-Met was supplemented to cows fed the high NFC diet. Insulin addition in vitro did not affect hepatic metabolism of propionate on d 1 postpartum. Overall, both the NFC content of the prepartum diet and Cr-Met had only modest effects on metabolic indices in this experiment. PMID- 18420632 TI - A model of phosphorus digestion and metabolism in the lactating dairy cow. AB - A dynamic, mechanistic, compartmental model of phosphorus (P) digestion and metabolism was constructed in the Advanced Continuous Simulation Language using conservation of mass principles and mass action kinetics. Phosphorus was assumed to exist in 3 forms: inorganic (Pi), phytic acid (Pp), and organic (excluding phytic acid; Po). All 3 forms were assumed to be present in the digestive tract with absorption of Pi into blood. Inputs to the model were total P intake; Pp, Po, and Pi as proportions of total P; milk yield; rate of salivation (fixed at 239 L/d); and rate of liquid passage from the rumen (fixed at 198 L/d). The model was fitted to 2 experiments from the literature. Derived parameters were well defined by the data. With a mean observed P intake of 75 g/d, total tract P digestibility was 38%. Phytic acid P digestibility in the rumen was 74%, with no additional Pp digestion in the lower tract. Inorganic P and Po digestibility in the lower tract were 48 and 89%, respectively. Flows of Po and Pi from the rumen were 2.4 and 3.0 times greater than intake, respectively. The increase in Po was apparently due to microbial growth. The increase in Pi arose primarily from secretion of Pi into the rumen via salivation where 65% of absorbed P was recycled to the rumen. Milk synthesis used 30% of absorbed Pi, and 1% was excreted in urine. This research suggested that the primary regulation points for maintaining blood P were bone deposition and resorption and absorption from the intestine. However, because bone P balance was related to both dietary P intake and ruminal phytase activity, it is critical to achieve a better understanding of phytate digestibility across several feeds if dietary P is to be reduced below current requirements. PMID- 18420633 TI - Frequent allocation of rotationally grazed dairy cows changes grazing behavior and improves productivity. AB - Twenty Holstein cows were blocked in 2 groups according to milk yield to evaluate the effect of frequency of allocation to new grazing plots on pasture intake, grazing behavior, rumen characteristics, and milk yield. The 2 treatments, daily allocation to 0.125-ha plots (1D) or allocation every 4 d to 0.5-ha plots (4D) of Lolium perenne L., were tested in a randomized block design (2 rotations with 3 or 4 measuring periods of 4 d each) with mixed model analysis accounting for repeated measures. There were no differences in the chemical composition of offered pasture and in pasture dry matter intake (DMI) between 1D and 4D. However, an interaction between treatment and rotation indicated a difference in pasture DMI between treatments during the first rotation (4D, 16.5 vs. 1D, 18.3 kg/d) but not during the second rotation (4D, 15.0 vs. 1D, 14.7 kg/d), possibly a result of a greater pasture mass in the first rotation. Grazing time (average 562 min/d) and ruminating time (average 468 min/d), observed using IGER graze recorders, were similar between treatments, but grazing time increased numerically (549 to 568 min/d), and ruminating time decreased linearly (471 to 450 min/d) within periods in the 4D treatment. Mean rumen pH (6.16 vs. 6.05) and rumen NH(3)-N concentration (113.7 vs. 90.1 mg/L) were higher in 4D than in 1D, and total volatile fatty acid (VFA) concentrations did not differ. Molar proportions of VFA, except butyrate, differed between treatments, causing the nonglucogenic to glucogenic VFA ratio to be greater in 4D than in 1D. Within days in the 4D treatment, the molar proportion of acetate increased and those of all other VFA decreased linearly. Rumen NH(3)-N concentration within the 4D treatment declined quadratically from 170.3 mg/L on d 1 to 80.7 mg/L on d 4. In contrast to rumen NH(3)-N concentration, milk urea content did not differ between treatments, but decreased quadratically from d 1 to 4 in the 4D treatment (from 26.7 to 20.7 mg/dL). Mean fat- and protein-corrected milk was greater in 1D than in 4D (23.5 vs. 22.8 kg/d), mainly due to a difference in milk yield (24.5 vs. 23.7 kg/d). Fat and protein content were slightly lower in the 1D than in the 4D treatment (3.66 vs. 3.76% and 3.28 vs. 3.34%, respectively). This study confirmed that increasing pasture allocation frequency from once every 4 d to every day improved milk production in grazing dairy cows, especially when offered pasture was high. PMID- 18420634 TI - Modeling the adequacy of dietary fiber in dairy cows based on the responses of ruminal pH and milk fat production to composition of the diet. AB - The main objective of this study was to develop practical models to assess and predict the adequacy of dietary fiber in high-yielding dairy cows. We used quantitative methods to analyze relevant research data and critically evaluate and determine the responses of ruminal pH and production performance to different variables including physical, chemical, and starch-degrading characteristics of the diet. Further, extensive data were used to model the magnitude of ruminal pH fluctuations and determine the threshold for the development of subacute ruminal acidosis (SARA). Results of this study showed that to minimize the risk of SARA, the following events should be avoided: 1) a daily mean ruminal pH lower than 6.16, and 2) a time period in which ruminal pH is <5.8 for more than 5.24 h/d. As the content of physically effective neutral detergent fiber (peNDF) or the ratio between peNDF and rumen-degradable starch from grains in the diet increased up to 31.2 +/- 1.6% [dry matter (DM) basis] or 1.45 +/- 0.22, respectively, so did the daily mean ruminal pH, for which a asymptotic plateau was reached at a pH of 6.20 to 6.27. This study also showed that digestibility of fiber in the total tract depends on ruminal pH and outflow rate of digesta from reticulorumen; thereby both variables explained 62% of the variation of fiber digestibility. Feeding diets with peNDF content up to 31.9 +/- 1.97% (DM basis) slightly decreased DM intake and actual milk yield; however, 3.5% fat-corrected milk and milk fat yield were increased, resulting in greater milk energy efficiency. In conclusion, a level of about 30 to 33% peNDF in the diet may be considered generally optimal for minimizing the risk of SARA without impairing important production responses in high-yielding dairy cows. In terms of improvement of the accuracy to assessing dietary fiber adequacy, it is suggested that the content of peNDF required to stabilize ruminal pH and maintain milk fat content without compromising milk energy efficiency can be arranged based on grain or starch sources included in the diet, on feed intake level, and on days in milk of the cows. PMID- 18420635 TI - Chitotriosidase activity in goat blood and colostrum. AB - Chitotriosidase (ChT) activity has not been investigated in ruminants, and therefore, we studied this activity in blood and colostrum of 25 pregnant goats and 60 goat kids. Blood samples were taken from pregnant goats at 3, 2, and 1 d prepartum; at partum; and at 1, 2, 3, and 4 d postpartum. Colostrum samples were obtained by machine-milking at partum and 1, 2, 3, and 4 d postpartum. Goat kid blood was collected at birth and every 7 d thereafter until goats kids were 56 d old. The ChT activity ranged from 2,368 to 3,350 nmol/ mL per hour in goat blood serum, and no statistical differences were detected through time. However, activity tended to decrease from 3 d prepartum to 2 d post-partum. Colostrum ChT activity was 3,912 nmol/mL per hour and 465 nmol/mL per hour on the day of delivery and 4 d postpartum, respectively. Colostrum ChT activity was significantly higher at partum than at any other time. The ChT activity in colostrum was significantly greater at 1 d postpartum than at 2, 3, and 4 d postpartum. Chitotriosidase activity did not differ in colostrum collected on d 2, 3, and 4 postpartum. Chitotriosidase activity in goat kid blood serum ranged from 2,664 to 9,231 nmol/mL per hour at birth and 49 d of life, respectively. Chitotriosidase activity in the blood serum increased with age: at birth, activity was significantly less than at 28, 35, 42, 49, and 56 d postpartum. The maximum ChT activity in blood serum was observed at 49 d postpartum. Activity in 49-d-old kids was significantly greater than that observed in kids at 0, 7, and 14 d postpartum. PMID- 18420636 TI - Technical note: a near-infrared reflectance spectroscopy technique to predict particle size of starch within corn silage. AB - Starch particle size characteristics of 81 diverse corn silage samples, which included 27 combinations of hybrid, planting dates, and harvest dates subjected to 3 different degrees of kernel processing, were determined via vertical shaking through 9 screens with nominal square apertures of 19.0, 13.2, 9.5, 6.7, 4.75, 3.35, 2.36, 1.18, and 0.6 mm and a pan. Starch content of dry matter remaining on each screen and on the pan for each corn silage was determined, and geometric mean particle size (GMPS, mum), starch particles(SP)/g, starch surface area (SSA, cm(2)/g), kernel processing score (KPS), % starch < 4.75 mm, and the percentage of total starch remaining on each screen of the vertical shakers were calculated. Near-infrared reflectance spectra were obtained by scanning 3 types of samples: 1 mm of dried ground corn silage; whole undried, unground corn silage; and undried, unground corn silage that passed through a 19-mm screen. Calibrations to predict GMPS, SP, SSA, and KPS characteristics of corn silage starch were attempted from each spectral origin. Calibrations to predict GMPS, SP, SSA, KPS, and the percentage of total starch retained on screens of the vertical shaker was unattainable (R(2) < 0.45) using spectra obtained from 1 mm of dried ground corn silage or whole undried, unground corn silage. However, reasonable near-infrared reflectance spectra equations (R(2) > 0.81) for GMPS, SSA, and KPS were attained using spectra from undried, unground corn silage that passed through a 19-mm screen. This technique holds promise as a rapid and efficient method to determine particle size characteristics of starch within corn silage. PMID- 18420637 TI - Short communication: salivary secretion during meals in lactating dairy cattle. AB - Four multiparous Holstein cows in midlactation were used in a 4 x 4 Latin square to evaluate whether source of forage influenced salivary secretion during eating in lactating dairy cows. The forages were allocated separately from the pelleted concentrates. Cows were offered 1 of 4 forages each period: barley silage, alfalfa silage, long-stemmed alfalfa hay, or chopped barley straw. Saliva secretion was measured during the morning meal by collecting masticates through the rumen cannula at the cardia of each cow. Rate of salivation (213 g/min) was not affected by forage source. However, the forage sources differed in eating rate (g of DM/min), which led to differences in ensalivation of forages (g of saliva/g of DM and g of saliva/g of NDF). On the basis of DM, ensalivation (g of saliva/g of DM) was greatest for straw (7.23) and similar for barley silage, alfalfa silage, and alfalfa hay (4.15, 3.40, and 4.34 g/g of DM, respectively). Higher ensalivation of straw could be accounted for by its higher neutral detergent fiber (NDF) content; ensalivation of NDF (g of saliva/g of NDF) was actually greatest for long-stemmed alfalfa hay (12.4) and similar for the other chopped forages (8.9). Cows consumed concentrate about 3 to 12 times faster than the various forages (DM basis), and ensalivation of concentrate was much lower (1.12 g of saliva/g of DM) than for forages. Feed characteristics such as particle size, DM, and NDF content affect salivary output during eating by affecting the eating rate. Slower eating rate and greater time spent eating may help prevent ruminal acidosis by increasing the total daily salivary secretion in dairy cows. PMID- 18420638 TI - Short communication: turbidity as an indicator of Escherichia coli presence in water troughs on cattle farms. AB - Studies have shown that water-drinking troughs are an important source of Escherichia coli infection on cattle farms, and a study was designed to provide farmers with an easy-to-use tool to monitor trough contamination and help determine when to empty and clean water troughs. A total of 164 water troughs were sampled on 33 cattle farms, and the on-farm turbidity tester results were found to be significantly correlated with laboratory turbidity results. Turbidity was associated with E. coli concentration, although the association was not linear. Emptying the troughs within a week of sampling was shown to reduce the turbidity score, but no linear association between time of emptying and E. coli concentration was discovered. A turbidity score of 4 was set as a cutoff point for when to clean a trough, yielding a sensitivity of 0.94 and a specificity of 0.03 for identifying a level of E. coli concentration that was more likely to contain E. coli O157 (>5,800 cfu/100 mL). PMID- 18420639 TI - Short communication: apoptosis regulates passive immune transfer in newborn kids. AB - Ten newborn kids were used to evaluate the relationship between apoptosis and passive immune transfer in neonatal enterocytes. Kids were slaughtered in groups of 2 at birth, 1, 2, 3, or 60 d postpartum, and samples of duodenal epithelium collected from each animal. Samples were fixed, dehydrated, and embedded in paraffin wax. Sections were assessed for apoptotic cells and immunostained for IgG. Our results suggest that IgG absorption is mediated by apoptotic enterocytes. Thus, delaying apoptosis may improve the success of passive immune transfer. PMID- 18420640 TI - Conformation of hind legs and lameness in Danish Holstein heifers. AB - Lameness causes major financial losses and welfare problems in dairy herds. Prevention of foot lesions may suffice in the majority of lameness cases. The objectives of this longitudinal study were to describe the dynamics and associations between abnormal hind leg conformation, asymmetric claws, lameness, and foot lesions in 122 Danish Holstein heifers from an average of 41 d before first calving until dry off or culling. The cattle were housed either in a free stall system with cubicles or in deep-bedded straw yard. The claws of all cattle were examined on up to 5 occasions. The associations between foot lesions, lameness, symmetry of the claws, shape of the dorsal toe-wall, and the conformation of the hind legs were examined statistically using mixed models. A large proportion (81%) of the heifers had cow-hocked conformation, with wide based stance, hocks together, and lateral rotation of the foot before calving, and 25% of the heifers had locomotion scores above 2, indicating signs of lameness, before calving. Our results indicate that lameness, abnormal conformation, and lesions acquired precalving persist throughout the first lactation. Thinner cows were more cow-hocked, and cow-hocked cows had a higher frequency of sole hemorrhages. Increased severity of white line lesion was associated with greater claw asymmetry. More severe lameness and sole hemorrhages were found in symmetric claws. PMID- 18420641 TI - Quantitative trait loci affecting calving traits in Danish Holstein cattle. AB - The objectives of this study were 1) to detect quantitative trait loci (QTL) affecting direct and maternal calving traits at first calving in the Danish Holstein population, 2) to distinguish between pleiotropic and linked QTL for chromosome regions affecting more than one trait, and 3) to detect QTL affecting stillbirth and calving difficulties but not calf size that could be used in selection to improve calving performance. Progeny-tested sons (2,297) were genotyped for 356 microsatellites in 34 grandsire families on all 29 autosomes. A total of 27 significant QTL on 17 chromosomes were detected using a between families linear regression model. For the direct calving traits, 4 QTL significantly affected calving difficulty, 5 QTL affected stillbirth, and 7 QTL affected calf size subjectively assessed by the farmer as a categorical trait. When the maternal components of the same traits were tested, there were significant effects of 3 QTL on calving difficulty, 6 QTL on stillbirth, and 2 QTL on calf size. The variance component mapping approach was used to estimate the relative posterior probability of linkage and pleiotropic models. The most probable model indicated a pleiotropic QTL on chromosome 12 and 25 and a linked QTL on chromosome 7 and 26. Chromosome 18 seemed to harbor a QTL with a pleiotropic effect on the direct calving traits and linked to maternal stillbirth. Markers on chromosomes 3, 4, 7, 10, 12, 18, 21, 24, 26, and 28 can be used to select new breeding candidates to produce daughters with more efficient calving performance. PMID- 18420642 TI - Extent of linkage disequilibrium in Holstein cattle in North America. AB - The success of fine-scale mapping and genomic selection depends mainly on the strength of linkage disequilibrium (LD) between markers and causal mutations. With Lewontin's measure of LD (known as D'), high levels of LD that extend over several million base pairs have been reported in livestock. However, this measure of LD can be strongly biased upward by small samples and by low allele frequencies. The aim of this study was to characterize the level and extent of LD in Holstein cattle in North America (Canada and the United States for purposes of this study) by using the squared correlation of the alleles at 2 loci (r(2)). The Affymetrix MegAllele GeneChip Bovine Mapping 10K single nucleotide polymorphism (SNP) array was used to genotype 821 bulls, from which 497 were used in the analysis of the extent of LD. A total of 5,564 SNP were used after filtering out SNP with more than 5% of Mendelian inconsistencies, with more than 20% missing genotypes, or with a minor allele frequency of less than 10%. Analysis of syntenic pairs revealed that useful LD (measured as r(2) > 0.3) occurred at distances shorter than 100 kb. Linkage disequilibrium decayed very rapidly, within a few hundred kilobase pairs. In addition, no substantial LD between unlinked loci was found. Using a sliding window analysis, we observed an irregular pattern of LD across the genome. These findings suggest that to capture useful LD, which is required for whole-genome fine mapping and genomic selection, a denser SNP map would be needed. PMID- 18420643 TI - Genetic parameters for tunisian holsteins using a test-day random regression model. AB - Genetic parameters of milk, fat, and protein yields were estimated in the first 3 lactations for registered Tunisian Holsteins. Data included 140,187; 97,404; and 62,221 test-day production records collected on 22,538; 15,257; and 9,722 first-, second-, and third-parity cows, respectively. Records were of cows calving from 1992 to 2004 in 96 herds. (Co)variance components were estimated by Bayesian methods and a 3-trait-3-lactation random regression model. Gibbs sampling was used to obtain posterior distributions. The model included herd x test date, age x season of calving x stage of lactation [classes of 25 days in milk (DIM)], production sector x stage of lactation (classes of 5 DIM) as fixed effects, and random regression coefficients for additive genetic, permanent environmental, and herd-year of calving effects, which were defined as modified constant, linear, and quadratic Legendre coefficients. Heritability estimates for 305-d milk, fat and protein yields were moderate (0.12 to 0.18) and in the same range of parameters estimated in management systems with low to medium production levels. Heritabilities of test-day milk and protein yields for selected DIM were higher in the middle than at the beginning or the end of lactation. Inversely, heritabilities of fat yield were high at the peripheries of lactation. Genetic correlations among 305-d yield traits ranged from 0.50 to 0.86. The largest genetic correlation was observed between the first and second lactation, potentially due to the limited expression of genetic potential of superior cows in later lactations. Results suggested a lack of adaptation under the local management and climatic conditions. Results should be useful to implement a BLUP evaluation for the Tunisian cow population; however, results also indicated that further research focused on data quality might be needed. PMID- 18420644 TI - Frequency and effect of the bovine acyl-CoA:diacylglycerol acyltransferase 1 (DGAT1) K232A polymorphism in Swedish dairy cattle. AB - Acyl-CoA:diacylglycerol acyltransferase 1 (DGAT1) is a key enzyme in triacylglycerol synthesis in the mammary gland, and the corresponding gene has emerged as a strong candidate for the variation in milk fat percentage. In this study, the allele frequencies and effects of the DGAT1 K232A variants in the Swedish dairy breeds Swedish Red and Swedish Holstein were investigated. A total of 239 cows, 143 of the Swedish Red breed and 96 of the Swedish Holstein breed, in the experimental herd at the Swedish University of Agricultural Sciences were genotyped for the DGAT1 polymorphism. The Swedish Red cows in the herd belonged to 1 of 2 selection lines with high or low milk fat percentage, respectively, but with similar high total milk energy production. The frequency of the K variant was found to be significantly greater in the high-fat line than in the low-fat line. The average frequency of the K variant in the 2 lines of the Swedish Red cows was 0.09 compared with 0.12 among the Swedish Holstein cows. Mixed model analysis was used to estimate the effect of the DGAT1 K232A polymorphism based on 16,866 test-day records for milk production traits. In accordance with previous studies, the most pronounced effects were found for fat and protein percentages and milk yield; and the K variant was associated with an increase in milk fat and protein percentages but less milk yield compared with the A variant. Less pronounced effects were found for yields of fat and protein for which the K variant was associated with greater fat yield but less protein yield. PMID- 18420645 TI - Milk fatty acid unsaturation: genetic parameters and effects of stearoyl-CoA desaturase (SCD1) and acyl CoA: diacylglycerol acyltransferase 1 (DGAT1). AB - With regard to human health aspects of milk fat, increasing the amount of unsaturated fatty acids in milk is an important selection objective. The cow's diet has an influence on the degree of unsaturation, but literature suggests that genetics also plays a role. To estimate genetic variation in milk fatty acid unsaturation indices, milk fatty acid composition of 1,933 Dutch Holstein Friesian heifers was measured and unsaturation indices were calculated. An unsaturation index represents the concentration of the unsaturated product proportional to the sum of the unsaturated product and the saturated substrate. Intraherd heritabilities were moderate, ranging from 0.23 +/- 0.07 for conjugated linoleic acid (CLA) index to 0.46 +/- 0.09 for C16 index. We genotyped the cows for the SCD1 A293V and DGAT1 K232A polymorphisms, which are known to alter milk fatty acid composition. Both genes explain part of the genetic variation in unsaturation indices. The SCD1 V allele is associated with lower C10, C12, and C14 indices, and with higher C16, C18, and CLA indices in comparison to the SCD1 A allele, with no differences in total unsaturation index. In comparison to the DGAT1 K allele, the DGAT1 A allele is associated with lower C10, C12, C14, and C16 indices and with higher C18, CLA, and total indices. We conclude that selective breeding can contribute to higher unsaturation indices, and that selective breeding can capitalize on genotypic information of both the SCD1 A293V and the DGAT1 K232A polymorphism. PMID- 18420646 TI - Economic impact of milk production in the State of New Mexico. AB - The goal of this study was to quantify the economic role of dairy farming in New Mexico and to identify its linkages with allied industries in terms of income, value added, and employment impacts. An input-output model was used to estimate the direct, indirect, and induced impacts of the dairy farm industry on the economy of New Mexico. The results showed that in 2005, New Mexico's dairy farm industry had a total economic impact of $1.98 billion and accounted for 14,313 jobs. Therefore, dairy farming in New Mexico had an output multiplier (income) of 1.92, a labor income multiplier of $248 thousand/$ million of gross sales, and an employment multiplier of 13.91 jobs/$ million of gross sales. Furthermore, the New Mexico dairy farms accounted for 13.1% of the total agricultural outputs, 20.5% of the agricultural jobs, 1.5% of total state economic activity, and $80 million in tax revenue. With the exception of Lea, Eddy, and Bernalillo counties, which are diversified, the dairy farms accounted for more than two-thirds of the agricultural outputs and for more than two-fifths of the agricultural employment in counties where dairy farms are concentrated. PMID- 18420647 TI - Effect of stocking rate on pasture production, milk production, and reproduction of dairy cows in pasture-based systems. AB - Ninety-four cows were randomly allocated to 1 of 5 stocking rates (2.2, 2.7, 3.1, 3.7, and 4.3 cows/ha) in a completely randomized design for 3 years. Herds were seasonal calving, with only minor differences in grazing management to optimize the profitability of each stocking rate (SR). Pasture production and quality data, milk and milk component data, and reproduction data were collected, averaged for SR treatment, and linear and quadratic contrasts on SR were evaluated. In addition, the Wilmink exponential model (y(t) = a + b x e((-0.05t) )+ c x t) was fitted to milk yield within lactation, and the parameters were averaged by SR treatment and analyzed as above. The median variation explained by the function for individual lactations was 84%. The amount of pasture grown tended to increase, and the quality of the pasture on offer increased linearly with increasing SR, reducing some of the negative impact of SR on the availability of pasture per cow. Milk production per cow declined linearly with increasing SR, although there was a tendency for most production variables to decline quadratically, with the negative effect of SR declining with increasing SR. The effect on milk production per cow was primarily because of a lower peak milk yield and a greater post-peak decline (less persistent milk profile), although a decline in lactation length with increasing SR was responsible for 24% of the effect of SR on milk yield. Milk production per hectare increased linearly with increasing SR, and there was only a small difference (approximately 3%/cow per ha) in the efficiency of converting feed dry matter into milk energy. Stocking rate did not affect reproductive success. The data are consistent with the need for a more robust measure of SR than cows per hectare because farms will differ in the genetic merit of their cows and in the potential to produce pasture. We introduce the concept of a comparative SR, whereby the carrying capacity of the farm is defined by the BW of the cows, the potential of the land to produce pasture, and the amount of supplement purchased (kg of BW/t of feed dry matter). The adoption of such a measure would facilitate the extrapolation and transfer of research findings among systems. PMID- 18420648 TI - Fertility and pregnancy outcome in women with congenital adrenal hyperplasia due to 21-hydroxylase deficiency. AB - BACKGROUND: Low pregnancy rate has been reported in women with congenital adrenal hyperplasia (CAH) and little information on pregnancy and children is known. METHODS: In a Swedish study, 62 adult women with CAH, aged 18-63 years, and 62 age-matched controls were followed-up. Medical records, including those concerning pregnancies and deliveries, were examined and the 21-hydroxylase genotype of patients was noted. All women answered a questionnaire concerning sexual and reproductive health including health of the children. RESULTS: Pregnancy and delivery rates were significantly lower in women with CAH (P < 0.001, P < 0.0056, respectively), and the severity of the 21-hydroxylase-mutation correlated with the reduced number of children born. More women with salt-wasting CAH were single and had not attempted pregnancy. Pregnancies were normal except for a significantly increased incidence of gestational diabetes in CAH patients (P < 0.0024). The children had normal birthweight and no malformations were observed. A later follow-up of the children showed a normal intellectual and social development. The sex ratio of the offspring differed significantly, with 25% boys in the CAH group compared with 56% among controls (P < 0.016). CAH women had more gynaecological morbidity during menopause. CONCLUSIONS: Pregnancy and delivery rates are reduced in women with CAH mainly due to psychosocial reasons. The outcome of children did not differ from controls. The unexpected sex ratio in children born to mothers with CAH warrants further research. PMID- 18420649 TI - Temporary pacing wire in the coronary sinus: a novel treatment of acute heart failure? AB - Treatment of acute heart failure relies initially on medical therapy. Patients can be considered for cardiac resynchronization therapy once they are able to lie flat for several hours. However, placement of a temporary pacing wire (TPW) into the coronary sinus may allow the patient to receive resynchronization therapy in the acute phase. We report a case of a patient who had a dramatic improvement of symptoms and blood pressure after a TPW was placed in the coronary sinus. PMID- 18420650 TI - Routine follow-up after pacemaker implantation: frequency, pacemaker programming and professionals in charge. AB - AIMS: To describe current evidence of the frequency, contents, and involved professionals of the routine follow-up visits in patients who have received a pacemaker (PM). METHODS AND RESULTS: The multicentre FOLLOWPACE study prospectively collected data during implantation and follow-up of 1526 patients who received a PM for the first time. A total of 4914 follow-up visits were studied. Mean follow-up was 394 days with a mean of 3.2 visits per patient. At all follow-up visits, the battery condition was tested in >93%, the stimulation threshold in >91%, and sensing in >87%. The pacemaker parameters as stimulation and sensing thresholds, lead impedances, and percentages of pacing remained stable over time, but these values did depend on the lead location, lead fixation, and pulse duration. The majority of PM (re-)programming was performed during implantation and/or shortly before hospital discharge (50%). PM re programming during follow-up was most frequently performed by the PM technician alone (95%). CONCLUSION: Crucial PM parameters are regularly checked. Re programming of PM parameters declined during the first year after PM implantation. The majority of PM checks were carried out by the PM technician, indicating the major influence of the allied professional on the quality and safety of the pacing therapy. PMID- 18420651 TI - Exploring the potential of pulmonary vein recordings: can they help elucidate mechanisms of paroxysmal atrial fibrillation? PMID- 18420652 TI - Pulmonary vein potentials in patients with and without atrial fibrillation. AB - BACKGROUND: Pulmonary vein (PV) potentials are invariably recordable at the PV ostia in patients with atrial fibrillation (AF) and delayed conduction around the PV ostia may play a role in the initiation and maintenance of AF. AIMS: To investigate the presence and extent of PV potentials in patients with and without AF. METHODS AND RESULTS: Circumferential catheter recordings at the PV ostia were obtained from 10 patients with paroxysmal AF and 9 with concealed Wolff-Parkinson White (WPW) syndrome without history of AF. Typical PV potential was defined as either rapid deflections that separated from atrial deflection with a time delay in-between, or multiphasic, continuous or fractionated potentials. The presence of PV potentials was verified during sinus rhythm and during atrial pacing at the distal coronary sinus for the left PVs or at the right atrial appendage for the right PVs. To quantify the extent in which the PV potentials were recordable, the number of PVs with typical PV potentials recordable was counted. The time interval from the onset to the end of the electrograms recordable at the PV ostium (A-PV interval) was measured, and the maximal and mean of this interval were obtained. Typical PV potentials were recorded in 31 of 34 PVs (91%) in patients with AF, but in 4 of 36 PVs (11%) in patients with concealed WPW. A narrow, biphasic or triphasic, potential was recorded in 3 of 34 PVs (9%) in patients with AF, but in 29 of 36 (81%) PVs in patients with concealed WPW. The maximal and mean A-PV intervals were significantly longer in patients with AF (71 +/- 24 and 49 +/- 13 ms) than in patients with concealed WPW syndrome (33 +/- 14 and 25 +/- 6 ms). CONCLUSION: In patients with AF, typical PV potentials with marked conduction time delay were almost invariably recordable at the PV ostium, but in patients without a history of AF, merely simple, narrow potentials were found. These findings support the involvement of conduction delay and re-entrant activities around the PV ostia in the genesis and/or perpetuation of AF. PMID- 18420653 TI - webPIPSA: a web server for the comparison of protein interaction properties. AB - Protein molecular interaction fields are key determinants of protein functionality. PIPSA (Protein Interaction Property Similarity Analysis) is a procedure to compare and analyze protein molecular interaction fields, such as the electrostatic potential. PIPSA may assist in protein functional assignment, classification of proteins, the comparison of binding properties and the estimation of enzyme kinetic parameters. webPIPSA is a web server that enables the use of PIPSA to compare and analyze protein electrostatic potentials. While PIPSA can be run with downloadable software (see http://projects.eml.org/mcm/software/pipsa), webPIPSA extends and simplifies a PIPSA run. This allows non-expert users to perform PIPSA for their protein datasets. With input protein coordinates, the superposition of protein structures, as well as the computation and analysis of electrostatic potentials, is automated. The results are provided as electrostatic similarity matrices from an all-pairwise comparison of the proteins which can be subjected to clustering and visualized as epograms (tree-like diagrams showing electrostatic potential differences) or heat maps. webPIPSA is freely available at: http://pipsa.eml.org. PMID- 18420654 TI - R-Coffee: a method for multiple alignment of non-coding RNA. AB - R-Coffee is a multiple RNA alignment package, derived from T-Coffee, designed to align RNA sequences while exploiting secondary structure information. R-Coffee uses an alignment-scoring scheme that incorporates secondary structure information within the alignment. It works particularly well as an alignment improver and can be combined with any existing sequence alignment method. In this work, we used R-Coffee to compute multiple sequence alignments combining the pairwise output of sequence aligners and structural aligners. We show that R Coffee can improve the accuracy of all the sequence aligners. We also show that the consistency-based component of T-Coffee can improve the accuracy of several structural aligners. R-Coffee was tested on 388 BRAliBase reference datasets and on 11 longer Cmfinder datasets. Altogether our results suggest that the best protocol for aligning short sequences (less than 200 nt) is the combination of R Coffee with the RNA pairwise structural aligner Consan. We also show that the simultaneous combination of the four best sequence alignment programs with R Coffee produces alignments almost as accurate as those obtained with R Coffee/Consan. Finally, we show that R-Coffee can also be used to align longer datasets beyond the usual scope of structural aligners. R-Coffee is freely available for download, along with documentation, from the T-Coffee web site (www.tcoffee.org). PMID- 18420655 TI - The YqfN protein of Bacillus subtilis is the tRNA: m1A22 methyltransferase (TrmK). AB - N(1)-methylation of adenosine to m(1)A occurs in several different positions in tRNAs from various organisms. A methyl group at position N(1) prevents Watson Crick-type base pairing by adenosine and is therefore important for regulation of structure and stability of tRNA molecules. Thus far, only one family of genes encoding enzymes responsible for m(1)A methylation at position 58 has been identified, while other m(1)A methyltransferases (MTases) remain elusive. Here, we show that Bacillus subtilis open reading frame yqfN is necessary and sufficient for N(1)-adenosine methylation at position 22 of bacterial tRNA. Thus, we propose to rename YqfN as TrmK, according to the traditional nomenclature for bacterial tRNA MTases, or TrMet(m(1)A22) according to the nomenclature from the MODOMICS database of RNA modification enzymes. tRNAs purified from a DeltatrmK strain are a good substrate in vitro for the recombinant TrmK protein, which is sufficient for m(1)A methylation at position 22 as are tRNAs from Escherichia coli, which natively lacks m(1)A22. TrmK is conserved in Gram-positive bacteria and present in some Gram-negative bacteria, but its orthologs are apparently absent from archaea and eukaryota. Protein structure prediction indicates that the active site of TrmK does not resemble the active site of the m(1)A58 MTase TrmI, suggesting that these two enzymatic activities evolved independently. PMID- 18420656 TI - Analysis of siRNA specificity on targets with double-nucleotide mismatches. AB - Although RNA interference as a tool for gene knockdown is a great promise for future applications, the specificity of small interfering RNA (siRNA)-mediated gene silencing needs to be thoroughly investigated. Most research regarding siRNA specificity has involved analysis of affected off-target genes instead of exploring the specificity of the siRNA itself. In this study we have developed an efficient method for generating a siRNA target library by combining a siRNA target validation vector with a nucleotide oligomix. We have used this library to perform an analysis of the silencing effects of a functional siRNA towards its target site with double-nucleotide mismatches. The results indicated that not only the positions of the mismatched base pair have an impact on silencing efficiency but also the identity of the mismatched nucleotide. Our data strengthen earlier observations of widespread siRNA off-target effects and shows that approximately 35% of the double-mutated target sites still causes knockdown efficiency of >50%. We also provide evidence that there may be substantial differences in knockdown efficiency depending on whether the mutations are positioned within the siRNA itself or in the corresponding target site. PMID- 18420657 TI - Developmentally programmed DNA splicing in Paramecium reveals short-distance crosstalk between DNA cleavage sites. AB - Somatic genome assembly in the ciliate Paramecium involves the precise excision of thousands of short internal eliminated sequences (IESs) that are scattered throughout the germline genome and often interrupt open reading frames. Excision is initiated by double-strand breaks centered on the TA dinucleotides that are conserved at each IES boundary, but the factors that drive cleavage site recognition remain unknown. A degenerate consensus was identified previously at IES ends and genetic analyses confirmed the participation of their nucleotide sequence in efficient excision. Even for wild-type IESs, however, variant excision patterns (excised or nonexcised) may be inherited maternally through sexual events, in a homology-dependent manner. We show here that this maternal epigenetic control interferes with the targeting of DNA breaks at IES ends. Furthermore, we demonstrate that a mutation in the TA at one end of an IES impairs DNA cleavage not only at the mutant end but also at the wild-type end. We conclude that crosstalk between both ends takes place prior to their cleavage and propose that the ability of an IES to adopt an excision-prone conformation depends on the combination of its nucleotide sequence and of additional determinants. PMID- 18420658 TI - Identification of soluble protein fragments by gene fragmentation and genetic selection. AB - We describe a new method, which identifies protein fragments for soluble expression in Escherichia coli from a randomly fragmented gene library. Inhibition of E. coli dihydrofolate reductase (DHFR) by trimethoprim (TMP) prevents growth, but this can be relieved by murine DHFR (mDHFR). Bacterial strains expressing mDHFR fusions with the soluble proteins green fluroscent protein (GFP) or EphB2 (SAM domain) displayed markedly increased growth rates with TMP compared to strains expressing insoluble EphB2 (TK domain) or ketosteroid isomerase (KSI). Therefore, mDHFR is affected by the solubility of fusion partners and can act as a reporter of soluble protein expression. Random fragment libraries of the transcription factor Fli1 were generated by deoxyuridine incorporation and endonuclease V cleavage. The fragments were cloned upstream of mDHFR and TMP resistant clones expressing soluble protein were identified. These were found to cluster around the DNA binding ETS domain. A selected Fli1 fragment was expressed independently of mDHFR and was judged to be correctly folded by various biophysical methods including NMR. Soluble fragments of the cell-surface receptor Pecam1 were also identified. This genetic selection method was shown to generate expression clones useful for both structural studies and antibody generation and does not require a priori knowledge of domain architecture. PMID- 18420659 TI - Recent-onset childhood arthritis--association with Streptococcus pyogenes in a population-based study. AB - OBJECTIVES: To assess the frequency of Streptococcus pyogenes in children with early arthritis, compare the characteristics in patients with post-streptococcal ReA (PSReA) with those in patients with other types of arthritis, and describe the occurrence of carditis in PSRA. PATIENTS: In a population-based Norwegian study, the physicians were asked to refer all children with suspected arthritis. The arthritis patients were followed up at 6 weeks, 6 months and 18 months. The presence of S. pyogenes was based on throat smear or antibodies. Echocardiography was performed in the patients with ARF or PSRA. RESULTS: Thirty-two (18%) of the 173 children with arthritis tested positive for S. pyogenes. The percentage of positive tests rose steadily with age and peaked at ages 8-11 (35%). Six weeks after admission arthritis was present in 33% of the PSRA patients, which was less frequent than in the juvenile idiopathic arthritis (JIA) patients (P < 0.001), but more frequent than in the transient arthritis patients (P = 0.012). Hip arthritis was more frequent and knee/ankle arthritis, ANA and HLA-B27 were less frequent in PSRA than in JIA (P < 0.001, P = 0.009 and P = 0.029, respectively). The PSRA patients were older than those with transient arthritis (P = 0.007). One child with ARF had carditis. CONCLUSIONS: Streptococcus pyogenes was present in 18% of children with arthritis. The patient characteristics, clinical presentation and early disease course in PSRA was different from that of JIA and transient arthritis. PMID- 18420660 TI - Effects of switching between anti-TNF therapies on HAQ response in patients who do not respond to their first anti-TNF drug. AB - OBJECTIVES: Small studies have shown an improvement in disease activity in patients with RA who have switched between anti-TNF therapies for reasons of inefficacy. However, it is not clear whether switching improves longer term outcomes, such as disability. This analysis compares changes in HAQ scores 1 yr following lack of response to a first anti-TNF based on subsequent treatment during that year. METHODS: Analysis was limited to RA patients with inefficacy to a first anti-TNF based on (i) clinician opinion and/or (ii) disease activity score in 28 joints and had an HAQ measured at time of non-response and 12 months later. Patients were classified into three groups based on treatment during the next 12 months: (i) continued anti-TNF despite non-response; (ii) stopped anti TNF with no further biologics; and (iii) switched to a second anti-TNF. Mean improvement in HAQ was compared among the groups using multivariable linear regression models. RESULTS: As of July 2006, 868 patients met the inclusion for this analysis. Four hundred and seventy-nine patients stopped anti-TNF of whom 331 switched to a second anti-TNF. Three hundred and eighty-nine continued treatment. Patients who continued and those who switched had improvements in HAQ over the 12 months, unlike patients who discontinued all biologic therapy. The best improvement was seen in those who switched [adjusted mean improvement in HAQ 0.15 (95% CI 0.26, 0.05)]. CONCLUSION: There is a significant improvement in HAQ in patients who switch to a second anti-TNF, providing an effective next choice of therapy for some patients who fail to respond to their first anti-TNF. PMID- 18420661 TI - Self monitoring of blood glucose in type 2 diabetes. PMID- 18420662 TI - Efficacy of self monitoring of blood glucose in patients with newly diagnosed type 2 diabetes (ESMON study): randomised controlled trial. AB - OBJECTIVES: To assess the effect of self monitoring of blood glucose concentrations on glycaemic control and psychological indices in patients with newly diagnosed type 2 diabetes mellitus. DESIGN: Prospective randomised controlled trial of self monitoring versus no monitoring (control). SETTING: Hospital diabetes clinics. PARTICIPANTS: 184 (111 men) people aged <70 with newly diagnosed type 2 diabetes referred to the participating diabetes clinics. Major exclusion criteria were secondary diabetes, insulin treatment, previous self monitoring of blood glucose. INTERVENTIONS: Participants were randomised to self monitoring or no monitoring (control) groups for one year with follow-up at three monthly intervals. Both groups underwent an identical structured core education programme. The self monitoring group received additional education on monitoring. MAIN OUTCOME MEASURES: Between group differences in HbA(1c), psychological indices, use of oral hypoglycaemic drugs, body mass index (BMI), and reported hypoglycaemia rates. RESULTS: 96 patients (55 men) were randomised to monitoring and 88 (56 men) to control. There were no baseline differences in mean (SD) age (57.7 (11.0) in monitoring group v 60.9 (11.5) in control group) or HbA(1c) (8.8 (2.1)% v 8.6 (2.3)%, respectively). Those in the monitoring group had a higher baseline BMI (34 (7) v 32 (6.2)). There were no significant differences between groups at any time point (12 months values given) in HbA(1c) (6.9 (0.8)% v 6.9 (1.2)%, P=0.69; 95% confidence interval for difference -0.25% to 0.38%), BMI (33.1 (6.4) v 31.8 (6.0); adjusted for baseline BMI, P=0.32), use of oral hypoglycaemic drugs, or reported incidence of hypoglycaemia. Monitoring was associated with a 6% higher score on the depression subscale of the well-being questionnaire (P=0.01). CONCLUSIONS: In patients with newly diagnosed type 2 diabetes self monitoring of blood glucose concentration has no effect on glycaemic control but is associated with higher scores on a depression subscale. TRIAL REGISTRATION: ISRCTN 49814766. PMID- 18420663 TI - Cost effectiveness of self monitoring of blood glucose in patients with non insulin treated type 2 diabetes: economic evaluation of data from the DiGEM trial. AB - OBJECTIVE: To assess the cost effectiveness of self monitoring of blood glucose alone or with additional training in incorporating the results into self care, in addition to standardised usual care for patients with non-insulin treated type 2 diabetes. DESIGN: Incremental cost utility analysis from a healthcare perspective. Data on resource use from the randomised controlled diabetes glycaemic education and monitoring (DiGEM) trial covered 12 months before baseline and 12 months of trial follow-up. Quality of life was measured at baseline and 12 months using the EuroQol EQ-5D questionnaire. SETTING: Primary care in the United Kingdom. PARTICIPANTS: 453 patients with non-insulin treated type 2 diabetes. INTERVENTIONS: Standardised usual care (control) compared with additional self monitoring of blood glucose alone (less intensive self monitoring) or with training in self interpretation of the results (more intensive self monitoring). MAIN OUTCOME MEASURES: Quality adjusted life years and healthcare costs (sterling in 2005-6 prices). RESULTS: The average costs of intervention were pound89 (euro113; $179) for standardised usual care, pound181 for less intensive self monitoring, and pound173 for more intensive self monitoring, showing an additional cost per patient of pound92 (95% confidence interval pound80 to pound103) in the less intensive group and pound84 ( pound73 to pound96) in the more intensive group. No other significant cost difference was detected between the groups. An initial negative impact of self monitoring on quality of life occurred, averaging -0.027 (95% confidence interval-0.069 to 0.015) for the less intensive self monitoring group and -0.075 (-0.119 to -0.031) for the more intensive group. CONCLUSIONS: Self monitoring of blood glucose with or without additional training in incorporating the results into self care was associated with higher costs and lower quality of life in patients with non insulin treated type 2 diabetes. In light of this, and no clinically significant differences in other outcomes, self monitoring of blood glucose is unlikely to be cost effective in addition to standardised usual care. TRIAL REGISTRATION: Current Controlled Trials ISRCTN47464659. PMID- 18420664 TI - Complaints against doctors. PMID- 18420665 TI - Darzi's review of quality of care in the NHS. PMID- 18420666 TI - Deaths in custody. PMID- 18420667 TI - Leprosy chemoprophylaxis: What's the need? PMID- 18420668 TI - Preventing falls among older people in hospital: Results are not surprising. PMID- 18420669 TI - Preventing falls among older people in hospital: Interpreting the null result. PMID- 18420670 TI - The woman question: Change in attitude is required. PMID- 18420671 TI - The woman question: Sweden is a good model. PMID- 18420672 TI - The woman question: Why is this news? PMID- 18420673 TI - Reducing NHS carbon footprint: Time for a culture change. PMID- 18420674 TI - HFEA will defend challenge from Christian group on hybrid embryo research. PMID- 18420676 TI - Wakefield admits fabricating events when he took children's blood samples. PMID- 18420677 TI - Germany liberalises law on stem cell research. PMID- 18420679 TI - US hospitals pass on most of the costs of errors. PMID- 18420680 TI - More than 10% of hospital deaths could be avoided, conference is told. PMID- 18420681 TI - Health authority knew its cancer testing was inaccurate two years before publicly revealing it, inquiry is told. PMID- 18420682 TI - Two doctors in Holland are to be prosecuted over use of complementary therapy. PMID- 18420683 TI - UK psychiatrists offer help to colleagues in Iraq. PMID- 18420684 TI - New combined pill to treat malaria is launched. PMID- 18420686 TI - US is urged to expand its healthcare workforce to cope with surge in number of elderly people. PMID- 18420687 TI - More than 26,000 Americans die each year because of lack of health insurance. PMID- 18420688 TI - Egyptian doctors who took part in forced HIV testing "violated medical ethics". PMID- 18420689 TI - Knowing me, knowing you. PMID- 18420690 TI - A general practitioner for every person in the world. PMID- 18420691 TI - Should we screen for abdominal aortic aneurysm? Yes. PMID- 18420692 TI - Should we screen for aortic aneurysm? No. PMID- 18420693 TI - Development of palliative care and legalisation of euthanasia: antagonism or synergy? PMID- 18420694 TI - Breast feeding. PMID- 18420695 TI - Confidentiality. PMID- 18420696 TI - The tale of an anorexic male medical student. PMID- 18420700 TI - Isolating the kinetics of facilitation in the mossy fibre pathway. PMID- 18420701 TI - Monitoring changes in membrane phosphatidylinositol 4,5-bisphosphate in living cells using a domain from the transcription factor tubby. AB - Phosphatidylinositol 4,5-bisphosphate (PtdIns(4,5)P(2)) is a key component in signal transduction, being a precursor to other signalling molecules and itself associated with roles in signal transduction and cell biology. Tubby is a membrane bound transcription factor whose dysfunction results in obesity in mice. It contains a domain that selectively binds PtdIns(4,5)P(2). We have investigated the use of a fluorescently tagged version of this domain to monitor changes in PtdIns(4,5)P(2) concentration in living cells and compared it to the pleckstrin homology domain of PLCdelta1. Our results show that selected mutants of this domain report receptor-mediated changes in cellular PtdIns(4,5)P(2). In contrast to the pleckstrin homology domain of PLCdelta1 it does not have a significant affinity for inositol 1,4,5-trisphosphate (IP(3)). Using a selected mutant, we examine the regulation of ATP-sensitive K(+) channels via a G(q/11)-coupled receptor. These experiments reveal a correlation between reporter translocation and the onset of current inhibition whilst the recovery of current after agonist removal is delayed when compared to the reporter. Furthermore our studies reveal the importance of Ca(2+) in determining the overall activity of phospholipase C in living cells. This probe may be valuable in examining changes in PtdIns(4,5)P(2) distinct from those of IP(3) in intact cells in a variety of physiological settings. PMID- 18420702 TI - Defective regulation of contractile function in muscle fibres carrying an E41K beta-tropomyosin mutation. AB - A novel E41K beta-tropomyosin (beta-Tm) mutation, associated with congenital myopathy and muscle weakness, was recently identified in a woman and her daughter. In both patients, muscle weakness was coupled with muscle fibre atrophy. It remains unknown, however, whether the E41K beta-Tm mutation directly affects regulation of muscle contraction, contributing to the muscle weakness. To address this question, we studied a broad range of contractile characteristics in skinned muscle fibres from the two patients and eight healthy controls. Results showed decreases (i) in speed of contraction at saturated Ca(2+) concentration (apparent rate constant of force redevelopment (k(tr)) and unloaded shortening speed (V(0))); and (ii) in contraction sensitivity to Ca(2+) concentration, in fibres from patients compared with controls, suggesting that the mutation has a negative effect on contractile function, contributing to the muscle weakness. To investigate whether these negative impacts are reversible, we exposed skinned muscle fibres to the Ca(2+) sensitizer EMD 57033. In fibres from patients, 30 mum of EMD 57033 (i) had no effect on speed of contraction (k(tr) and V(0)) at saturated Ca(2+) concentration but (ii) increased Ca(2+) sensitivity of contraction, suggesting a potential therapeutic approach in patients carrying the E41K beta-Tm mutation. PMID- 18420703 TI - Cyclothiazide: a subunit-specific inhibitor of GABAC receptors. AB - We tested the effects of cyclothiazide (CTZ), an agent used to block desensitization of AMPA-type glutamate receptors, on heterologously expressed GABA(C) receptors formed by homomeric rho subunits. CTZ inhibition of GABA(C) receptors was subunit specific; it produced a dose-dependent reduction of the GABA-elicited current on homomeric rho2 receptors with an IC(50) of about 12 microm, but had no significant effect on homomeric rho1 receptors. This differential sensitivity was attributable to a single amino acid located on the second transmembrane domain of the rho subunits. Mutating the residue at this position from serine to proline on the rho2 subunit eliminated CTZ sensitivity, whereas switching proline to serine on the rho1 subunit made the receptor CTZ sensitive. The inhibitory properties of CTZ were consistent with its action as a channel blocker on the receptors formed by rho2 subunits. The effect showed a small degree of voltage dependence, and was due mainly to a non-competitive mechanism that reduced the maximum response elicited by GABA. In addition, the prominent membrane current rebound when co-application of GABA and CTZ was terminated suggests that the binding site for CTZ on the GABA(C) receptor is distinct from that for GABA, and that CTZ acts as a non-competitive antagonist on the GABA(C) receptor. CTZ inhibited the open channel of the GABA(C) receptor with a time constant of about 0.4 s, but the kinetics were approximately 10-fold slower when GABA is absent. The ability of CTZ to interact with various types of neurotransmitter receptors indicates that the drug has multiple actions in the CNS. PMID- 18420704 TI - Adaptive changes of the locomotor pattern and cutaneous reflexes during locomotion studied in the same cats before and after spinalization. AB - Descending supraspinal inputs exert powerful influences on spinal reflex pathways in the legs. Removing these inputs by completely transecting the spinal cord changes the state (i.e. the configuration of the spinal circuitry) of the locomotor network and undoubtedly generates a reorganization of reflex pathways. To study changes in reflex pathways after a complete spinalization, we recorded spinal reflexes during locomotion before and after a complete transection of the spinal cord at the 13th thoracic segment in cats. We chronically implanted electrodes in three cats, to record electromyography (EMG) in several hindlimb muscles and around the left tibial (Tib) nerve at the ankle to elicit reflexes during locomotion before and after spinalization in the same cat. Control values of kinematics, EMGs and reflexes were obtained during intact locomotion for 33-60 days before spinalization. After spinalization, cats were trained 3-5 times a week on a motorized treadmill. Recordings resumed once a stable spinal locomotion was achieved (26-43 days), with consistent plantar foot placement and full hindquarter weight support without perineal stimulation. Changes in Tib nerve reflex responses after spinalization in the same cat during locomotion were found in all muscles studied and were often confined to specific phases of the step cycle. The most remarkable change was the appearance of short-latency excitatory responses in some ipsilateral ankle extensors during stance. Short-latency excitatory responses in the ipsilateral tibialis anterior were increased during stance, whereas in other flexors such as semitendinosus and sartorius, increases were mostly confined to swing. Longer-latency excitatory responses in ipsilateral flexors were absent or reduced. Responses evoked in limb muscles contralateral to stimulation were generally increased throughout the step cycle. These reflex changes after spinalization provide important clues regarding the functional reorganization of reflex pathways during spinal locomotion. PMID- 18420705 TI - Presynaptic (Type III) cells in mouse taste buds sense sour (acid) taste. AB - Taste buds contain two types of cells that directly participate in taste transduction - receptor (Type II) cells and presynaptic (Type III) cells. Receptor cells respond to sweet, bitter and umami taste stimulation but until recently the identity of cells that respond directly to sour (acid) tastants has only been inferred from recordings in situ, from behavioural studies, and from immunostaining for putative sour transduction molecules. Using calcium imaging on single isolated taste cells and with biosensor cells to identify neurotransmitter release, we show that presynaptic (Type III) cells specifically respond to acid taste stimulation and release serotonin. By recording responses in cells isolated from taste buds and in taste cells in lingual slices to acetic acid titrated to different acid levels (pH), we also show that the active stimulus for acid taste is the membrane-permeant, uncharged acetic acid moiety (CH(3)COOH), not free protons (H(+)). That observation is consistent with the proximate stimulus for acid taste being intracellular acidification, not extracellular protons per se. These findings may also have implications for other sensory receptors that respond to acids, such as nociceptors. PMID- 18420706 TI - How do tonic glutamatergic synapses evade receptor desensitization? AB - Photoreceptor output synapses are the best known tonic chemical synapses in the nervous system, in which glutamate is continuously released in darkness, activating AMPA/kainate receptors in postsynaptic neurons. It has been shown that glutamate receptors in certain types of second-order retinal cells are largely desensitized in darkness, leading to small postsynaptic currents and reduced response dynamic ranges. Here we show that the tonic glutamatergic synapses between photoreceptors and rod-dominated hyperpolarizing bipolar cells (HBC(R)s) in the salamander retina evade postsynaptic receptor desensitization by using (1) multiple invaginating ribbon junctions as releasing sites for low-frequency, synchronized multiquantal release at each site; and (2) the GluR4 AMPA receptors as the postsynaptic receptors. The multiquantal events exhibit faster decay time than the GluR4 receptor desensitization time constant and therefore self desensitization is minimized, and the average inter-event duration in darkness is much longer than the GluR4 desensitization recovery time and thus mutual desensitization is avoided. Consequently, the HBC(R)s are not desensitized in darkness, allowing light signals to be encoded by the full operating range of the glutamate-gated postsynaptic currents. Our study illustrates for the first time how a tonic glutamatergic synapse avoids postsynaptic receptor desensitization, a strategy that may be shared by many other synapses in the nervous system that need extended operation capacity. PMID- 18420707 TI - Phosphocreatine as an energy source for actin cytoskeletal rearrangements during myoblast fusion. AB - Myoblast fusion is essential for muscle development, postnatal growth and muscle repair after injury. Recent studies have demonstrated roles for actin polymerization during myoblast fusion. Dynamic cytoskeletal assemblies directing cell-cell contact, membrane coalescence and ultimately fusion require substantial cellular energy demands. Various energy generating systems exist in cells but the partitioning of energy sources during myoblast fusion is unknown. Here, we demonstrate a novel role for phosphocreatine (PCr) as a spatiotemporal energy buffer during primary mouse myoblast fusion with nascent myotubes. Creatine treatment enhanced cell fusion in a creatine kinase (CK)-dependent manner suggesting that ATP-consuming reactions are replenished through the PCr/CK system. Furthermore, selective inhibition of actin polymerization prevented myonuclear addition following creatine treatment. As myotube formation is dependent on cytoskeletal reorganization, our findings suggest that PCr hydrolysis is coupled to actin dynamics during myoblast fusion. We conclude that myoblast fusion is a high-energy process, and can be enhanced by PCr buffering of energy demands during actin cytoskeletal rearrangements in myoblast fusion. These findings implicate roles for PCr as a high-energy phosphate buffer in the fusion of multiple cell types including sperm/oocyte, trophoblasts and macrophages. Furthermore, our results suggest the observed beneficial effects of oral creatine supplementation in humans may result in part from enhanced myoblast fusion. PMID- 18420708 TI - Role of sarcolemmal ATP-sensitive K+ channels in the regulation of sinoatrial node automaticity: an evaluation using Kir6.2-deficient mice. AB - The role of cardiac sarcolemmal ATP-sensitive K+ (K(ATP)) channels in the regulation of sinoatrial node (SAN) automaticity is not well defined. Using mice with homozygous knockout (KO) of the Kir6.2 (a pore-forming subunit of cardiac K(ATP) channel) gene, we investigated the pathophysiological role of K(ATP) channels in SAN cells during hypoxia. Langendorff-perfused mouse hearts were exposed to hypoxic and glucose-free conditions (hypoxia). After 5 min of hypoxia, sinus cycle length (CL) was prolonged from 207 +/- 10 to 613 +/- 84 ms (P < 0.001) in wild-type (WT) hearts. In Kir6.2 KO hearts, CL was slightly prolonged from 198 +/- 17 to 265 +/- 32 ms. The CL of spontaneous action potentials of WT SAN cells, recorded in the current-clamp mode, was markedly prolonged from 410 +/ 56 to 605 +/- 108 ms (n = 6, P < 0.05) with a decrease of the slope of the diastolic depolarization (SDD) after the application of the K+ channel opener pinacidil (100 microm). Pinacidil induced a glibenclamide (1 microm)-sensitive outward current, which was recorded in the voltage-clamp mode, only in WT SAN cells. During metabolic inhibition by 2,4-dinitrophenol, CL was prolonged from 292 +/- 38 to 585 +/- 91 ms (P < 0.05) with a decrease of SDD in WT SAN cells but not in Kir6.2 KO SAN cells. Diastolic Ca2+ concentration, measured by fluo-3 fluorescence, was decreased in WT SAN cells but increased in Kir6.2 KO SAN cells after short-term metabolic inhibition. In conclusion, the present study using Kir6.2 KO mice indicates that, during hypoxia, activation of sarcolemmal K(ATP) channels in SAN cells inhibits SAN automaticity, which is important for the protection of SAN cells. PMID- 18420709 TI - Ethinyl oestradiol administration in women suppresses synthesis of collagen in tendon in response to exercise. AB - Women are at greater risk than men of sustaining certain kinds of injury and diseases of collagen-rich tissues. To determine whether a high level of oestradiol has an acute influence on collagen synthesis in tendons at rest and in response to exercise, one-legged kicking exercise was performed for 60 min at 67% of maximum power by healthy, young oral contraceptive (OC) users when circulating synthetic (ethinyl) oestradiol was high (n = 11, HE-OC) and compared to similar women who had never used OCs when circulating endogenous oestrogen was low (n = 12, LE-NOC). Interstitial fluid was collected 24 h post-exercise through microdialysis catheters placed anterior to the patellar tendon in both legs and subsequently analysed for the amino-terminal propeptide of type I collagen (PINP), a marker of tendon collagen synthesis. To determine the long-term effect of OC usage, patellar tendon cross-sectional area (CSA) was measured by magnetic resonance imaging (MRI). A lower exercise-induced increase in tendon collagen synthesis was observed in HE-OC than in LE-NOC (DeltaPINP (mean +/- s.e.m.) 1.5 +/- 5.3 versus 24.2 +/- 9.4 ng ml(-1), P < 0.05). Furthermore, serum and the interstitial peritendinous tissue concentrations of insulin-like growth factor I (IGF-I) and IGF-binding proteins showed a reduced bioavailability in HE-OC compared with results in LE-NOC. No difference in patellar tendon CSA was observed between groups. In conclusion, the selective increase in tendon collagen synthesis in LE-NOC but not HE-OC 24 h post-exercise is consistent with the hypothesis that oestradiol inhibits exercise-induced collagen synthesis in human tendon. The mechanism behind this is either a direct effect of oestradiol, or an indirect effect via a reduction in levels of free IGF-I. However, the data did not indicate any long-term effect on tendon size associated with chronic OC use. PMID- 18420710 TI - Developmental changes in the human GH receptor and its signal transduction pathways. AB - We previously reported the presence of functional human GH receptors (hGHRs) in the human fetal hepatocyte (FH) as early as the first trimester. Interestingly, fetal serum levels of hGH are in the acromegalic range, yet certain hGH-dependent factors are expressed at very low levels (IGF-I, IGF-binding protein-3), suggesting that fetal liver has limited responsiveness to hGH. To determine whether this is due to the fetal tissue levels of hGHR or factors in the hGH/hGHR axis that might influence hGHR function, we compared hGHR isoforms and downstream signaling proteins in FH versus human adult liver (HAL). Immunoprecipitation/immunoblotting (IB) analyses found similar precursor and mature hGHR forms while RT-PCR assays of truncated (T) hGHR(1-279), dominant negative for the full-length (FL) receptor, showed similar T/FL mRNA ratios in FH and HAL. IB demonstrated that Janus kinase (JAK) 2, signal transducers and activators of transcription (STAT(1, 3, 5A/B)), and suppressors of cytokine signaling (SOCS(1, 2, 3, cytokine-inducible SH2-containing protein (CIS))) proteins were detectable in all FH and HAL tested (12 weeks of fetal age to 60 years); the levels were similar (STAT5B) or lower (JAK2/STAT1/STAT3/STAT5A: 38 53%, SOCS/CIS: 58-76%) in FH compared with HAL. Our studies to date demonstrate that, during hepatocyte development, hGHR levels are lower in the fetal cells but the hGHR isoforms, including the relative amount of truncated versus FL, remain unchanged. The JAK2/STAT/SOCS signaling molecules are present in the FH as early as the first trimester. However, they are generally at <50% level in postnatal liver. These data suggest that low expression of both hGHR and major hGHR signaling components may explain the limited responsiveness of the fetal cells to the high circulating levels of hGH. PMID- 18420711 TI - The relative value of operon predictions. AB - For most organisms, computational operon predictions are the only source of genome-wide operon information. Operon prediction methods described in literature are based on (a combination of) the following five criteria: (i) intergenic distance, (ii) conserved gene clusters, (iii) functional relation, (iv) sequence elements and (v) experimental evidence. The performance estimates of operon predictions reported in literature cannot directly be compared due to differences in methods and data used in these studies. Here, we survey the current status of operon prediction methods. Based on a comparison of the performance of operon predictions on Escherichia coli and Bacillus subtilis we conclude that there is still room for improvement. We expect that existing and newly generated genomics and transcriptomics data will further improve accuracy of operon prediction methods. PMID- 18420712 TI - Interrelation of inflammation and APP in sIBM: IL-1 beta induces accumulation of beta-amyloid in skeletal muscle. AB - Distinct interrelationships between inflammation and beta-amyloid-associated degeneration, the two major hallmarks of the skeletal muscle pathology in sporadic inclusion body myositis (sIBM), have remained elusive. Expression of markers relevant for these pathomechanisms were analysed in biopsies of sIBM, polymyositis (PM), dermatomyositis (DM), dystrophic and non-myopathic muscle as controls, and cultured human myotubes. By quantitative PCR, a higher upregulation was noted for the mRNA-expression of CXCL-9, CCL-3, CCL-4, IFN-gamma, TNF-alpha and IL-1 beta in sIBM muscle compared to PM, DM and controls. All inflammatory myopathies displayed overexpression of degeneration-associated markers, yet only in sIBM, expression of the mRNA of amyloid precursor protein (APP) significantly and consistently correlated with inflammation in the muscle and mRNA-levels of chemokines and IFN-gamma. Only in sIBM, immunohistochemical analysis revealed that inflammatory mediators including IL-1 beta co-localized to beta-amyloid depositions within myofibres. In human myotubes, exposure to IL-1 beta caused upregulation of APP with subsequent intracellular aggregation of beta-amyloid. Our data suggest that, in sIBM muscle, production of high amounts of pro inflammatory mediators specifically induces beta-amyloid-associated degeneration. The observations may help to design targeted treatment strategies for chronic inflammatory disorders of the skeletal muscle. PMID- 18420713 TI - Are muscles mechanically independent? PMID- 18420714 TI - Resistance training induces qualitative changes in muscle morphology, muscle architecture, and muscle function in elderly postoperative patients. AB - Although the negative effects of bed rest on muscle strength and muscle mass are well established, it still remains a challenge to identify effective methods to restore physical capacity of elderly patients recovering from hospitalization. The present study compared different training regimes with respect to muscle strength, muscle fiber size, muscle architecture, and stair walking power in elderly postoperative patients. Thirty-six patients (60-86 yr) scheduled for unilateral hip replacement surgery due to hip osteoarthritis were randomized to either 1) resistance training (RT: 3/wk x 12 wk), 2) electrical stimulation (ES: 1 h/day x 12 wk), or 3) standard rehabilitation (SR: 1 h/day x 12 wk). All measurements were performed at baseline, at 5 wk and 12 wk postsurgery. After 12 wk of resistance training, maximal dynamic muscle strength increased by 30% at 60 degrees /s (P < 0.05) and by 29% at 180 degrees /s (P < 0.05); muscle fiber area increased for type I (+17%, P < 0.05), type IIa (+37%, P < 0.05), and type IIx muscle fibers (+51%, P < 0.05); and muscle fiber pennation angle increased by 22% and muscle thickness increased by 15% (P < 0.05). Furthermore, stair walking power increased by 35% (P < 0.05) and was related to the increase in type II fiber area (r = 0.729, P < 0.05). In contrast, there was no increase in any measurement outcomes with electrical stimulation and standard rehabilitation. The present study is the first to demonstrate the effectiveness of resistance training to induce beneficial qualitative changes in muscle fiber morphology and muscle architecture in elderly postoperative patients. In contrast, rehabilitation regimes based on functional exercises and neuromuscular electrical stimulation had no effect. The present data emphasize the importance of resistance training in future rehabilitation programs for elderly individuals. PMID- 18420715 TI - Single bouts of exercise affect albumin redox state and carbonyl groups on plasma protein of trained men in a workload-dependent manner. AB - The purpose of this study was to investigate the effect of single bouts of exercise at three different intensities on the redox state of human serum albumin (HSA) and on carbonyl groups on protein (CP) concentrations in plasma. Trained men [n = 44, maximal oxygen consumption (Vo(2max)): 55 +/- 5 ml.kg(-1).min(-1), nonsmokers, 34 +/- 5 years of age] from a homogenous population, volunteers from a police special forces unit, were randomly assigned to perform on a cycle ergometer either at 70% (n = 14), 75% (n = 14), or 80% (n = 16) of Vo(2max) for 40 min. Blood was collected before exercise, immediately after the exercise test (IE), and 30 min after each test (30M) and 30 h after each test (30H). The reduced fraction of HSA, human mercaptalbumin (HMA), decreased at all three exercise intensities IE and 30M, returning to preexercise values by 30H (P < 0.05). HMA was primarily oxidized to its reversible fraction human nonmercaptalbumin 1 (HNA1). CP concentrations increased at 75% of Vo(2max) IE and 30M with a tendency (P < 0.1) and at 80% Vo(2max) IE and 30M significantly, returning to preexercise concentrations by 30H (P < 0.01). These results indicate that the HSA redox system in plasma is activated after a single bout of cycle ergometer exercise at 70% Vo(2max) and 40 min duration. The extent of the HSA modification increased with exercise intensity. Oxidative protein damage, as indicated by CP, was only significantly increased at 80% Vo(2max) intensity in this homogenous cohort of trained men. PMID- 18420716 TI - Some aspects of clinical relevance in the maturation of respiratory control in infants. AB - Two reflex mechanisms important for survival are discussed. Brain stem and cardiovascular mechanisms that are responsible for recovery from severe hypoxia (autoresuscitation) are important for survival in acutely hypoxic infants and adults. Failure of this mechanism may be important in sudden infant death syndrome (SIDS), because brain stem-mediated hypoxic gasping is essential for successful autoresuscitation and because SIDS infants appear to attempt to autoresuscitate just before death. A major function of another mechanism is to protect the airway from fluid aspiration. The various components of the laryngeal chemoreflex (LCR) change during maturation. The LCR is an important cause of prolonged apneic spells in infants. Consequently, it also may have a role in causing SIDS. Maturational changes and/or inadequacy of this reflex may be responsible for pulmonary aspiration and infectious pneumonia in both children and adults. PMID- 18420717 TI - Computational assessment of airway wall stiffness in vivo in allergically inflamed mouse models of asthma. AB - Allergic inflammation is known to cause airway hyperresponsiveness in mice. However, it is not known whether inflammation affects the stiffness of the airway wall, which would alter the load against which the circumscribing smooth muscle shortens when activated. Accordingly, we measured the time course of airway resistance immediately following intravenous methacholine injection in acutely and chronically allergically inflamed mice. We estimated the effective stiffness of the airway wall in these animals by fitting to the airway resistance profiles a computational model of a dynamically narrowing airway embedded in elastic parenchyma. Effective airway wall stiffness was estimated from the model fit and was found not to change from control in either the acute or chronic inflammatory groups. However, the acutely inflamed mice were hyperresponsive compared with controls, which we interpret as reflecting increased delivery of methacholine to the airway smooth muscle through a leaky pulmonary endothelium. These results support the notion that acutely inflamed BALB/c mice represent an animal model of functionally normal airway smooth muscle in a transiently abnormal lung. PMID- 18420718 TI - Adenosine induces a cholinergic tracheal reflex contraction in guinea pigs in vivo via an adenosine A1 receptor-dependent mechanism. AB - Adenosine induces dyspnea, cough, and airways obstruction in asthma, a phenomenon that also occurs in various sensitized animal models in which a neuronal involvement has been implicated. Although adenosine has been suggested to activate cholinergic nerves, the precise mechanism has not been established. In the present study, the adenosine A(1) receptor agonist N(6)-cyclopentyladenosine (CPA) induced a cholinergic reflex, causing tracheal smooth muscle contraction that was significantly inhibited by the adenosine A(1) receptor antagonist 8 cyclopentyl-1,3-dipropylxanthine (DPCPX; 100 microg/kg) (P < 0.05) in anesthetized animals. Furthermore, the adenosine A(2) agonist 2-p-(2 carboxyethyl) phenethylamino-5'-N-ethylcarboxamidoadenosine (CGS-21680) induced a small reflex, whereas the A(3) selective agonist N(6)-(3-iodobenzyl)-5'-N methylcarbamoyladenosine (IB-MECA) was without effect. The tracheal reflex induced by CPA was also inhibited by recurrent nerve ligation or muscarinic receptor blockade (P < 0.001), indicating that a cholinergic neuronal mechanism of action accounted for this response. The cholinergic reflex in response to aerosolized CPA was significantly greater in passively sensitized compared with naive guinea pigs (P < 0.01). Chronic capsaicin treatment, which inhibited sensory nerve function, failed to inhibit CPA-induced reflex tracheal contractions in passively sensitized guinea pigs, although the local anesthetic lidocaine inhibited CPA-induced tracheal contractions. The effects of CPA on the reflex response was not dependent on the release of histamine from tissue mast cells or endogenous prostaglandins as shown by the lack of effect of the histamine H(1) receptor antagonist pyrilamine (1 mg/kg) or the cyclooxygenase inhibitor meclofenamic acid (3 mg/kg), respectively. In conclusion, activation of pulmonary adenosine A(1) receptors can stimulate cholinergic reflexes, and these reflexes are increased in allergic guinea pigs. PMID- 18420719 TI - Jump exercise during remobilization restores integrity of the trabecular architecture after tail suspension in young rats. AB - Three-dimensional trabecular architecture was investigated in the femora of tail suspended young growing rats, and the effects of jump exercise during remobilization were examined. Five-week-old male Wistar rats (n = 35) were randomly assigned to five body weight-matched groups: tail-suspended group (SUS; n = 7); sedentary control group for SUS (S(CON); n = 7); spontaneous recovery group after tail suspension (S+R(CON), n = 7); jump exercise group after tail suspension (S+R(JUM); n = 7); and age-matched control group for S+R(CON) and S+R(JUM) without tail suspension and exercise (S(CON)+R(CON); n = 7). Rats in SUS and S(CON) were killed immediately after tail suspension for 14 days. The jump exercise protocol consisted of 10 jumps/day, 5 days/wk, and jump height was 40 cm. Bone mineral density (BMD) of the femur and three-dimensional trabecular bone architecture at the distal femoral metaphysis were measured. Tail suspension induced a 13.6% decrease in total femoral BMD (P < 0.001) and marked deterioration of trabecular architecture. After 5 wk of free remobilization, femoral BMD, calf muscle weight, and body weight returned to age-matched control levels, but trabeculae remained thinner and less connected. On the other hand, S+R(JUM) rats showed significant increases in trabecular thickness, number, and connectivity compared with S+R(CON) rats (62.8, 31.6, and 24.7%, respectively; P < 0.05), and these parameters of trabecular architecture returned to the levels of S(CON)+R(CON). These results indicate that suspension-induced trabecular deterioration persists after remobilization, but jump exercise during remobilization can restore the integrity of trabecular architecture and bone mass in the femur in young growing rats. PMID- 18420720 TI - Peripheral oxygen transport and utilization in rats following continued selective breeding for endurance running capacity. AB - Untrained rats selectively bred for either high (HCR) or low (LCR) treadmill running capacity previously demonstrated divergent physiological traits as early as the seventh generation (G7). We asked whether continued selective breeding to generation 15 (G15) would further increase the divergence in skeletal muscle capillarity, morphometry, and oxidative capacity seen previously at G7. At G15, mean body weight was significantly lower (P < 0.001) in the HCR rats (n = 11; 194 +/- 3 g) than in LCR (n = 12; 259 +/- 9 g) while relative medial gastrocnemius muscle mass was not different (0.23 +/- 0.01 vs. 0.22 +/- 0.01% total body weight). Normoxic (Fi(O(2)) = 0.21) Vo(2max) was 50% greater (P < 0.001) in HCR despite the lower absolute muscle mass, and skeletal muscle O(2) conductance (measured in hypoxia; Fi(O(2)) = 0.10) was 49% higher in HCR (P < 0.001). Muscle oxidative enzyme activities were significantly higher in HCR (citrate synthase: 16.4 +/- 0.4 vs. 14.0 +/- 0.6; beta-hydroxyacyl-CoA dehydrogenase: 5.2 +/- 0.2 vs. 4.2 +/- 0.2 mmol.kg(-1).min(-1)). HCR rats had approximately 36% more total muscle fibers and also 36% more capillaries in the medial gastrocnemius. Because average muscle fiber area was 35% smaller, capillary density was 36% higher in HCR, but capillary-to-fiber ratio was the same. Compared with G7, G15 HCR animals showed 38% greater total fiber number with an additional 25% decrease in mean fiber area. These data suggest that many of the skeletal muscle structural and functional adaptations enabling greater O(2) utilization in HCR at G7 continue to progress following additional selective breeding for endurance capacity. However, the largest changes at G15 relate to O(2) delivery to skeletal muscle and not to the capacity of skeletal muscle to use O(2). PMID- 18420721 TI - Postnatal changes in the expression of serotonin 2A receptors in various brain stem nuclei of the rat. AB - Previously, we reported a critical period [around postnatal day (P) 12-13 in the rat] in respiratory network development when distinct neurochemical, metabolic, and physiological changes occur. Since serotonin 2A (5-HT(2A)) receptors play an important role in respiratory modulation, we hypothesized that they may undergo developmental adjustments during the critical period. Semi-quantitative immunohistochemical analyses were conducted in labeled neurons in a number of brain stem nuclei with or without known respiratory functions from P2 to P21 in rats. Our data indicate that the expressions of 5-HT(2A) receptors in neurons of the pre-Botzinger complex, the nucleus ambiguus, and the hypoglossal nucleus were maintained within a relatively narrow range between P2 and P21, with a dip at P3 P4 and a significant reduction only at P12. This change was not observed in the nonrespiratory cuneate nucleus. These results suggest that reduced expressions of 5-HT(2A) receptors at P12 contributes to neurochemical imbalance within brain stem respiratory nuclei at that time and may be involved in decreased hypoxic ventilatory response at this critical period of development. PMID- 18420722 TI - Contribution of male sex, age, and obesity to mechanical instability of the upper airway during sleep. AB - Male sex, obesity, and age are risk factors for obstructive sleep apnea, although the mechanisms by which these factors increase sleep apnea susceptibility are not entirely understood. This study examined the interrelationships between sleep apnea risk factors, upper airway mechanics, and sleep apnea susceptibility. In 164 (86 men, 78 women) participants with and without sleep apnea, upper airway pressure-flow relationships were characterized to determine their mechanical properties [pharyngeal critical pressure under hypotonic conditions (passive Pcrit)] during non-rapid eye movement sleep. In multiple linear regression analyses, the effects of body mass index and age on passive Pcrit were determined in each sex. A subset of men and women matched by body mass index, age, and disease severity was used to determine the sex effect on passive Pcrit. The passive Pcrit was 1.9 cmH(2)O [95% confidence interval (CI): 0.1-3.6 cmH(2)O] lower in women than men after matching for body mass index, age, and disease severity. The relationship between passive Pcrit and sleep apnea status and severity was examined. Sleep apnea was largely absent in those individuals with a passive Pcrit less than -5 cmH(2)O and increased markedly in severity when passive Pcrit rose above -5 cmH(2)O. Passive Pcrit had a predictive power of 0.73 (95% CI: 0.65-0.82) in predicting sleep apnea status. Upper airway mechanics are differentially controlled by sex, obesity, and age, and partly mediate the relationship between these sleep apnea risk factors and obstructive sleep apnea. PMID- 18420723 TI - Physical activity and bone development during childhood: insights from animal models. AB - Animal studies illustrate greater structural and material adaptations of growing bone to exercise than in adult bones but do not define effective training regimes to optimize bone strength in children. Controlled loading studies in turkey, rat, or mouse bones have revealed mechanisms of mechanotransduction and loading characteristics that optimize the modeling response to applied strains. Insights from these models reveal that static loads do not play a role in mechanotransduction and that bone formation is threshold driven and dependent on strain rate, amplitude, and partitioning of the load. That is, only a few cycles of loading are required at any time to elicit an adaptive response, and distributed bouts of loading, incorporating rest periods, are more osteogenic than single sessions of long duration. These parameters of loading have been translated into feasible public health interventions that exploit the insights gained from animal experiments to achieve adaptive responses in children and adolescents. Studies manipulating estrogen receptors (ER) in mice also demonstrate that skeletal sensitivity to loading during the peripubertal period is due to a direct regulation of mechanotransduction pathways by ER, and not just a simple enhancement of cell activity already marshaled by the hypothalamic pituitary axis. Unfortunately, because the rate and timing of growth in small animals are completely different from those in humans, these models can be poor tools to elucidate periods during growth in youths, during which the skeleton is more sensitive to loading. However, there are insights from studies of human growth that can improve the interpretation of data from such studies of growth and development in animals. PMID- 18420724 TI - The endocrine response and substrate utilization during exercise in children and adolescents. AB - Adolescence is a time of rapid growth caused by significant changes in hormone levels. For many, it is also a time of increased physical activity and sport that places a large demand on energy reserves. Exercise is known to cause perturbations in endocrine and metabolic systems in children and adolescents, yet careful characterization of these responses is only now being conducted. It does not appear that prepubertal youth have a different muscle composition than adults. However, these youth do have a lower anaerobic capacity and a greater reliance on aerobic metabolism during activity. Prepubertal adolescents may have an immature glucose regulatory system that influences glycemic regulation at the onset of moderate exercise. During heavy exercise, muscle and blood lactate levels are lower in children than in adults and there is a greater reliance on fat as fuel. The exercise intensity that causes maximal fat oxidation rate and the relative rate of fat oxidation decreases as adolescents develop through puberty. The mechanism for the attenuated lipid utilization with the advancement of puberty, and the impact that this may have on body composition, are unknown. Surprisingly, prepubertal adolescents have relatively high rates of exogenous glucose oxidation, perhaps because of their smaller endogenous carbohydrate reserves. Further study is needed to determine the optimal exogenous carbohydrate feeding regimen for peak performance in adolescence. Studies are also needed to determine whether physical activity, at an intensity targeted to maximize fat oxidation, help to lower body adiposity in overweight youth. PMID- 18420725 TI - Recurrent trigeminal neuralgia: long term outcome of repeat gamma knife radiosurgery. AB - OBJECTIVE: To date, the efficacy and safety of repeat radiosurgery (RS) for trigeminal neuralgia (TN) is based mainly on short term results. METHODS: Between 1994 and 2006, 93 patients were treated by RS for TN at the Department of Neurosurgery, Graz, Austria. 22 patients underwent repeat gamma knife radiosurgery (GKRS) a mean of 18.8 months after the initial treatment. The mean dose for repeat treatment was 74.3 Gy. Pain outcome was rated using the Barrow Neurological Institute (BNI) Pain Intensity Scale and facial numbness according to the BNI Facial Numbness Scale. RESULTS: Mean follow-up after repeat RS was 5.4 years. Pain relief was noted in 72.7% (16/22) of patients; six patients had a second pain recurrence after a mean of 9.3 months and underwent medical, alternative and/or further RS. One patient was lost to follow-up. BNI pain scale evaluation for 21 patients indicated improvement in 76.2% (16/21) of cases without medication (BNI I and II). Facial numbness was recorded in 73.7% (14/19) but in only one was it classified as bothersome. CONCLUSIONS: Long term observation of repeat GKRS for TN showed good pain relief in more than two-thirds of patients. Despite a high percentage of facial numbness, most likely attributable to the higher delivered dose, repeat RS can still be regarded as safe. However, further studies are needed to determine an optimised treatment protocol. PMID- 18420726 TI - Utility of the stroke-thrombolytic predictive instrument. AB - OBJECTIVE: The goal of this study was to evaluate the utility of the stroke thrombolytic predictive instrument (s-TPI) in predicting clinical outcome in patients with acute ischaemic stroke treated with intravenous tissue plasminogen activator (t-PA). METHODS: The study assessed the external validity of the s-TPI in 301 consecutive stroke patients treated with intravenous t-PA. Clinical outcome was measured with the modified Rankin scale (mRs) at 3 months. The study used the s-TPI to calculate probabilities of a good outcome (mRs 0-1) and a poor outcome (mRs 5-6). We compared these probabilities with the observed outcome using receiver-operator characteristics (ROC) curves and calibration curves. Subgroup analyses for different onset-to-treatment time windows were performed. RESULTS: According to the s-TPI, the mean predicted probability of a good and a poor outcome in the validation cohort were 0.45 and 0.17. The area under the ROC curves were 0.80 (4.5-hour time window), 0.82 (3-hour time window) and 0.77 (3 4.5 hours time window) for predicting good outcome, and 0.78 (4.5 hours), 0.80 (3 hours) and 0.74 (3-4.5 hours) for predicting poor outcome. Calibration curves revealed a slight overestimation of probabilities of a good outcome and underestimation of probabilities of a poor outcome. CONCLUSIONS: The s-TPI appears to be reasonably valid for predicting outcome after t-PA treatment in daily practice, although a slight overestimation of a good and underestimation of a poor outcome was observed. PMID- 18420727 TI - Intractable hiccup and nausea in neuromyelitis optica with anti-aquaporin-4 antibody: a herald of acute exacerbations. AB - BACKGROUND: Intractable hiccup and nausea (IHN) are unique symptoms in neuromyelitis optica (NMO). Recent studies have strongly suggested that the pathogenesis of NMO is closely associated with anti-aquaporin-4 (AQP4) antibody. However, clinical implications of IHN and the relationship with anti-AQP4 antibody remain unknown. METHODS: The past medical records of 35 patients with seropositivity for anti-AQP4 antibody were reviewed. We also followed the titres of anti-AQP4 antibody in a patient with NMO, who had newly developed IHN. RESULTS: Of the 35 patients, 15 patients (43%) had episodes of IHN. There was a total of 35 episodes of IHN in these 15 patients and, of the 35 episodes, hiccup was seen in 23 episodes (66%) and nausea was seen in 28 episodes (80%). The IHN frequently preceded (54%) or accompanied (29%) myelitis or optic neuritis. The IHN was often preceded by an episode of viral infection. The titres of anti-AQP4 antibody were remarkably increased when the intractable hiccup appeared in a case. CONCLUSIONS: IHN could be a clinical marker for the early phase of an exacerbation. Careful observation may be needed when INH is seen in patients with NMO, and the early initiation of the treatment could prevent subsequent neurological damage. PMID- 18420728 TI - Quantitative assessment of hand disability by the Nine-Hole-Peg test (9-HPT) in cervical spondylotic myelopathy. PMID- 18420729 TI - A novel NGFB point mutation: a phenotype study of heterozygous patients. AB - OBJECTIVE: A family with neurological findings similar to hereditary sensory and autonomic neuropathy type V having a point mutation in the nerve growth factor beta (NGFB) gene was recently described. The homozygous genotype gives disabling symptoms. The purpose of the present study was to evaluate the symptoms in heterozygous patients. METHODS: 26 patients heterozygous for the NGFB mutation (12 men, mean age 50 (13-90) years) were examined clinically and answered a health status questionnaire, including the Michigan Neuropathy Screening Instrument (MNSI). 28 relatives (15 men, mean age 44 (15-86) years) without the mutation served as controls in the clinical examination part. 23 of the heterozygotes were examined neurophysiologically and six heterozygous patients underwent a sural nerve biopsy. RESULTS: The heterozygous phenotype ranged from eight patients with Charcot arthropathy starting in adult age and associated with variable symptoms of neuropathy but without complete insensitivity to pain, anhidrosis or mental retardation, to 10 symptom free patients. There was no difference in MNSI between the young heterozygous cases (<55 years old) and the controls. Six of 23 heterozygous patients had impaired cutaneous thermal perception and 11 of 23 had signs of carpal tunnel syndrome. Sural nerve biopsies showed a moderate reduction of both small myelinated (Adelta) and unmyelinated (C) fibres. No apparent correlation of small fibre reduction to symptoms was found. CONCLUSIONS: The NGFB mutation in its heterozygous form results in a milder disease than in homozygotes, with a variable clinical picture, ranging from asymptomatic cases to those with Charcot arthropathy appearing in adult age. Particularly age, but perhaps lifestyle factors also, may influence the development of clinical polyneuropathy. PMID- 18420730 TI - Standardization of C-peptide measurements. AB - BACKGROUND: C-peptide is a marker of insulin secretion in diabetic patients. We assessed within- and between-laboratory imprecision of C-peptide assays and determined whether serum calibrators with values assigned by mass spectrometry could be used to harmonize C-peptide results. METHODS: We sent 40 different serum samples to 15 laboratories, which used 9 different routine C-peptide assay methods. We also sent matched plasma samples to another laboratory for C-peptide analysis with a reference mass spectrometry method. Each laboratory analyzed 8 of these samples in duplicate on each of 4 days to evaluate within- and between-day imprecision. The same 8 samples were also used to normalize the results for the remaining samples to the mass spectrometry reference method. RESULTS: Within- and between-run CVs ranged from <2% to >10% and from <2% to >18%, respectively. Normalizing the results with serum samples significantly improved the comparability among laboratories and methods. After normalization, the differences among laboratories in mean response were no longer statistically significant (P = 0.24), with least-squares means of 0.93-1.02. CONCLUSIONS: C peptide results generated by different methods and laboratories do not always agree, especially at higher C-peptide concentrations. Within-laboratory imprecision also varied, with some methods giving much more consistent results than others. These data show that calibrating C-peptide measurement to a reference method can increase comparability between laboratories. PMID- 18420731 TI - Cell-free plasma DNA as a predictor of outcome in severe sepsis and septic shock. AB - BACKGROUND: Increased concentrations of cell-free DNA have been found in plasma of septic and critically ill patients. We investigated the value of plasma DNA for the prediction of intensive care unit (ICU) and hospital mortality and its association with the degree of organ dysfunction and disease severity in patients with severe sepsis. METHODS: We studied 255 patients with severe sepsis or septic shock. We obtained blood samples on the day of study inclusion and 72 h later and measured cell-free plasma DNA by real-time quantitative PCR assay for the beta globin gene. RESULTS: Cell-free plasma DNA concentrations were higher at admission in ICU nonsurvivors than in survivors (median 15 904 vs 7522 genome equivalents [GE]/mL, P < 0.001) and 72 h later (median 15 176 GE/mL vs 6758 GE/mL, P = 0.004). Plasma DNA values were also higher in hospital nonsurvivors than in survivors (P = 0.008 to 0.009). By ROC analysis, plasma DNA concentrations had moderate discriminative power for ICU mortality (AUC 0.70 0.71). In multiple regression analysis, first-day plasma DNA was an independent predictor for ICU mortality (P = 0.005) but not for hospital mortality. Maximum lactate value and Sequential Organ Failure Assessment score correlated independently with the first-day plasma DNA in linear regression analysis. CONCLUSIONS: Cell-free plasma DNA concentrations were significantly higher in ICU and hospital nonsurvivors than in survivors and showed a moderate discriminative power regarding ICU mortality. Plasma DNA concentration was an independent predictor for ICU mortality, but not for hospital mortality, a finding that decreases its clinical value in severe sepsis and septic shock. PMID- 18420732 TI - Aspirin responsiveness in healthy volunteers measured with multiple assay platforms. AB - BACKGROUND: We evaluated the sensitivity, precision, and concordance of 4 assays designed to detect aspirin responsiveness or resistance. METHODS: Twenty-nine healthy laboratory volunteers took 80 mg aspirin for 7 days, and a subset of volunteers took 325 mg aspirin for an additional 7 days. We measured platelet function by light transmission aggregometry with arachidonic acid, PFA-100, and VerifyNow. PFA-100 and VerifyNow assays were performed in duplicate to assess method imprecision. Some volunteers had samples taken within 2-4 h of the final dose of aspirin and again within 20-24 h of the final dose. We measured urinary 11-dehydro-thromboxane B(2) at baseline and after 80 or 325 mg aspirin. RESULTS: No volunteers were nonresponsive to aspirin therapy as measured by the PFA-100. One of 29 participants demonstrated lack of response to aspirin as measured by VerifyNow and urinary 11-dehydro-thromboxane B(2); 2 of 29 demonstrated lack of response as measured by light transmission aggregometry. Imprecision was <10% for the PFA-100 and VerifyNow. Concordance was high (>90%) between all assays. Neither aspirin dose (80 vs 325 mg) nor timing between final dose of aspirin and blood draw (2-4 vs 20-24 h) affected any of the assays. CONCLUSIONS: Light transmission aggregometry, PFA-100, VerifyNow, and urinary 11-dehydro-thromboxane B(2) are all sensitive to the effects of aspirin in healthy individuals. Variables such as aspirin dose, timing between final dose of aspirin and blood collection, and imprecision do not affect the ability of the assays to detect aspirin effect on platelet function. PMID- 18420733 TI - Detection of a thalassemic alpha-chain variant (Hemoglobin Groene Hart) by reversed-phase liquid chromatography. AB - BACKGROUND: Hemoglobin (Hb) Groene Hart [alpha119 (H2)Pro-->Ser (alpha1)], also known as Hb Bernalda, is a nondeletional alpha-thalassemic Hb variant that is frequent in southern Italy and North Africa. This variant is not supposed to be produced in the erythrocytes of carriers. The alpha-thalassemic behavior of this variant has been explained as an impaired interaction between the alpha-globin chain and the alpha-Hb-stabilizing protein. METHODS: To separate globin chains, we developed a modified reversed-phase liquid chromatography (RPLC) procedure that uses acetonitrile-water solvents containing up to 3 mL/L trifluoroacetic acid. After RPLC, we characterized the isolated globin chains by electrospray ionization (ESI) mass spectrometry (MS) and analyzed their tryptic peptides with matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) MS and nano-LC-ESI-MS/MS. RESULTS: RPLC detected an abnormal peak with a retention time substantially greater than that of the wild-type alpha(A)-globin chain. We identified this variant as Hb Groene Hart and found it in the hemolysates of 11 unrelated patients (1 homozygote, 9 heterozygotes, and 1 heterozygote associated with the -alpha(3.7) deletion). These patients possessed abnormal hematologic features suggesting an alpha-thalassemia phenotype. Molecular modeling suggested that the increase in hydrophobicity was due to opening of the GH interhelical segment following replacement of amino acid residue 119 with a nonhelix breaker residue. CONCLUSIONS: This method allows the detection of Hb variants at low concentrations, and adjusting the composition of the organic solvents enables the method to identify Hb variants with large changes in hydrophobicity. PMID- 18420734 TI - Presence of arylsulfatase A and sulfogalactosylglycerolipid in mouse ovaries: localization to the corpus luteum. AB - Arylsulfatase A (AS-A) is a lysosomal enzyme, which catalyzes the desulfation of certain sulfogalactolipids, including sulfogalactosylglycerolipid (SGG), a molecule implicated in cell adhesion. In this report, immunocytochemistry revealed the selective presence of AS-A in the corpus luteum of mouse ovaries. Immunoblotting indicated that mouse corpus luteum AS-A had a molecular mass of 66 kDa, similar to AS-A of other tissues. Corpus luteum AS-A was active, capable of desulfating the artificial substrate, p-nitrocatechol sulfate, at the optimum pH of five. To understand further the role of AS-A in female reproduction, levels of AS-A were determined during corpus luteum development in pseudopregnant mice and during luteolysis after cessation of pseudopregnancy. Immunocytochemistry, immunoblotting and desulfation activity showed that AS-A expression was evident at the onset of pseudopregnancy in the newly formed corpora lutea, and its level increased steadily during gland development. The increase in the expression and activity of AS-A continued throughout luteolysis after the decrease in serum progesterone levels. We also observed the selective presence of SGG on the luteal cell surface in developed corpora lutea, as shown by immunofluorescence of mouse ovary sections as well as high-performance thin-layer chromatography of lipids isolated from mouse and pig corpora lutea. The identity of the "SGG" band on the thin layer silica plate was further validated by electrospray ionization mass spectrometry. Significantly, SGG disappeared in regressing corpora lutea. Therefore, lysosomal AS-A may be involved in cell-surface remodeling during luteolysis by desulfating SGG after its endocytosis and targeting to the lysosome. PMID- 18420735 TI - Prolactin promotes the secretion of active cathepsin D at the basal side of rat mammary acini. AB - Cathepsin D (CD), a lysosomal aspartic protease present in mammary tissue and milk in various molecular forms, is also found in the incubation medium of mammary acini in molecular forms that are proteolytically active on prolactin at a physiological pH. Because prolactin controls the vesicular traffic in mammary cells, we studied, in vivo and in vitro, its effects on the polarized transport and secretion of various forms of CD in the rat mammary gland. CD accumulated in vesicles not involved in endocytosis in the basal region of cells. Prolactin increased this accumulation and the release of endosomal active single-chain CD at the basal side of acini. The CD-mediated proteolysis of prolactin, leading to the antiangiogenic 16-kDa form, at a physiological pH, was observed only in conditioned medium but not milk. These data support the novel concept that an active molecular form of CD, secreted at the basal side of the mammary epithelium, participates in processing blood-borne prolactin outside the cell, this polarized secretion being controlled by prolactin itself. PMID- 18420736 TI - Tissue-specific actions of the Ept1, Ept2, Ept6, and Ept9 genetic determinants of responsiveness to estrogens in the female rat. AB - Ept1, Ept2, Ept6, and Ept9 are quantitative trait loci mapped in crosses between the ACI and Copenhagen (COP) rat strains as genetic determinants of responsiveness of the pituitary gland to estrogens. We have developed four congenic rat strains, each of which carries, on the genetic background of the ACI rat strain, alleles from the COP rat strain that span one of these quantitative trait loci. Relative to the female ACI rats, female ACI.COP-Ept1 rats exhibited reduced responsiveness to 17beta-estradiol (E2) in the pituitary gland, as evidenced by quantification of pituitary mass and circulating prolactin, and in the mammary gland, as evidenced by reduced susceptibility to E2-induced mammary cancer. The ACI.COP-Ept2 rat strain exhibited reduced responsiveness to E2 in the pituitary gland but did not differ from the ACI strain in regard to susceptibility to E2-induced mammary cancer. Interestingly, female Ept2 congenic rats exhibited increased responsiveness to E2 in the thymus, as evidenced by enhanced thymic atrophy. The ACI.COP-Ept6 rat strain exhibited increased responsiveness to E2 in the pituitary gland, which was associated with a qualitative phenotype suggestive of enhanced pituitary vascularization. The ACI.COP-Ept9 rat strain exhibited reduced responsiveness to E2 in the anterior pituitary gland, relative to the ACI rat strain. Neither Ept6 nor Ept9 impacted responsiveness to E2 in the mammary gland or thymus. These data indicate that each of these Ept genetic determinants of estrogen action is unique in regard to the tissues in which it exerts its effects and/or the direction of its effect on estrogen responsiveness. PMID- 18420737 TI - Notch inhibits osteoblast differentiation and causes osteopenia. AB - Notch receptors are determinants of cell fate decisions. To define the role of Notch in the adult skeleton, we created transgenic mice overexpressing the Notch intracellular domain (NICD) under the control of the type I collagen promoter. First-generation transgenics were small and osteopenic. Bone histomorphometry revealed that NICD caused a decrease in bone volume, secondary to a reduction in trabecular number; osteoblast and osteoclast number were decreased. Low fertility of founder mice and lethality of young pups did not allow the complete establishment of transgenic lines. To characterize the effect of Notch overexpression in vitro, NICD was induced in osteoblasts and stromal cells from Rosa(notch) mice, in which a STOP cassette flanked by lox(P) sites is upstream of NICD, by transduction with an adenoviral vector expressing Cre recombinase (Cre) under the control of the cytomegalovirus (CMV) promoter (Ad-CMV-Cre). NICD impaired osteoblastogenesis and inhibited Wnt/beta-catenin signaling. To determine the effects of notch1 deletion in vivo, mice in which notch1 was flanked by lox(P) sequences (notch1(loxP/loxP)) were mated with mice expressing Cre recombinase under the control of the osteocalcin promoter. Conditional null notch1 mice had no obvious skeletal phenotype, possibly because of rescue by notch2; however, 1-month-old females exhibited a modest increase in osteoclast surface and eroded surface. Osteoblasts from notch1(loxP/loxP) mice, transduced with Ad-CMV-Cre and transfected with Notch2 small interfering RNA, displayed increased alkaline phosphatase activity. In conclusion, Notch signaling in osteoblasts causes osteopenia and impairs osteo-blastogenesis by inhibiting the Wnt/beta-catenin pathway. PMID- 18420738 TI - Characterization of nuclear import of the domain-specific androgen receptor in association with the importin alpha/beta and Ran-guanosine 5'-triphosphate systems. AB - Androgen induces androgen receptor (AR) nuclear import, which allows AR to act as a transcriptional factor and ultimately leads to biological activity. However, the mechanism of AR translocation to the nucleus is still unclear. In the present study, we assessed the nuclear import abilities of each domain of AR and their mechanisms related to Ran and importin alpha/beta using green fluorescent protein real-time imaging. The localization of AR to the nucleus in the absence and presence of ligands was dependent upon a complex interplay of the amino terminal transactivation domain (NTD), the DNA binding domain (DBD), and the ligand binding domain (LBD). NTD and DBD showed ligand-independent nuclear import ability, whereas LBD had ligand-dependent transport. In addition, AR deletion mutant lacking DBD was distributed in the cytoplasm regardless of ligand existence, suggesting that the remaining domains, NTD and LBD, are responsible for AR cytoplasmic localization. Cotransfection with a dominant negative form of Ran dramatically inhibited the nuclear import of all AR domains, and a dominant negative form of importin alpha prevented AR and DBD import. Importin beta knockdown strongly blocked DBD import. These results indicate that there are two additional nuclear localization signals (NLSs) in the NTD and LBD, and there are distinct pathways used to attain domain-specific AR nuclear import: the NLS of DBD is Ran and importin alpha/beta-dependent, whereas the NLSs of NTD and LBD are Ran dependent but importin alpha/beta-independent. Our data suggest that the nuclear import of AR is regulated by the interplay between each domain of the AR. PMID- 18420739 TI - Polychlorinated biphenyls (Aroclor 1254) do not uniformly produce agonist actions on thyroid hormone responses in the developing rat brain. AB - Thyroid hormone (TH) is essential for normal brain development, and polychlorinated biphenyls (PCBs) are known to interfere with TH action in the developing brain. Thus, it is possible that the observed neurotoxic effects of PCB exposure in experimental animals and humans are mediated in part by their ability to interfere with TH signaling. PCBs may interfere with TH signaling by reducing circulating levels of TH, acting as TH receptor analogs, or both. If PCBs act primarily by reducing serum TH levels, then their effects should mimic those of low TH. In contrast, if PCBs act primarily as TH agonists in the developing brain, then they should mimic the effect of T(4) in hypothyroid animals. We used a two-factor design to test these predictions. Both hypothyroidism (Htx) and/or PCB treatment reduced serum free and total T(4) on postnatal d 15. However, only Htx increased pituitary TSHbeta expression. RC3/neurogranin expression was decreased by Htx and increased by PCB treatment. In contrast, Purkinje cell protein-2 expression was reduced in hypothyroid animals and restored by PCB treatment. Finally, PCB treatment partially ameliorated the effect of Htx on the thickness of the external granule layer of the cerebellum. These studies demonstrate clearly that PCB exposure does not mimic the effect of low TH on several important TH-sensitive measures in the developing brain. However, neither did PCBs mimic T(4) in hypothyroid animals on all end points measured. Thus, PCBs exert a complex action on TH signaling in the developing brain. PMID- 18420740 TI - Caudal brainstem processing is sufficient for behavioral, sympathetic, and parasympathetic responses driven by peripheral and hindbrain glucagon-like peptide-1 receptor stimulation. AB - The effects of peripheral glucagon like peptide-1 receptor (GLP-1R) stimulation on feeding, gastric emptying, and energetic responses involve vagal transmission and central nervous system processing. Despite a lack of studies aimed at determining which central nervous system regions are critical for the GLP-1R response production, hypothalamic/forebrain processing is regarded as essential for these effects. Here the contribution of the caudal brainstem to the control of food intake, core temperature, heart rate, and gastric emptying responses generated by peripheral delivery of the GLP-1R agonist, exendin-4 (Ex-4), was assessed by comparing responses of chronic supracollicular decerebrate (CD) rats to those of pair-fed intact control rats. Responses driven by hindbrain intracerebroventricular (fourth i.c.v) delivery of Ex-4 were also evaluated. Intraperitoneal Ex-4 (1.2 and 3.0 microg/kg) suppressed glucose intake in both CD rats (5.0+/-1.2 and 4.4+/-1.1 ml ingested) and controls (9.4+/-1.5 and 7.7+/-0.8 ml ingested), compared with intakes after vehicle injections (13.1+/-2.5 and 13.2+/-1.7 ml ingested, respectively). Hindbrain ventricular Ex-4 (0.3 microg) also suppressed food intake in CD rats (4.7+/-0.6 ml ingested) and controls (11.0+/-2.9 ml ingested), compared with vehicle intakes (9.3+/-2.1 and 19.3+/-4.3 ml ingested, respectively). Intraperitoneal Ex-4 (0.12, 1.2, 2.4 microg/kg) reduced gastric emptying rates in a dose-related manner similarly for both CD and control rats. Hypothermia followed ip and fourth i.c.v Ex-4 in awake, behaving controls (0.6 and 1.0 C average suppression) and CD rats (1.5 and 2.5 C average suppression). Intraperitoneal Ex-4 triggered tachycardia in both control and CD rats. Results demonstrate that caudal brainstem processing is sufficient for mediating the suppression of intake, core temperature, and gastric emptying rates as well as tachycardia triggered by peripheral GLP-1R activation and also hindbrain-delivered ligand. Contrary to the literature, hypothalamic/forebrain processing and forebrain-caudal brainstem communication is not required for the observed responses. PMID- 18420741 TI - Comparative analysis of the uterine and mammary gland effects of drospirenone and medroxyprogesterone acetate. AB - The role of progestins in combined hormone therapy is the inhibition of uterine epithelial cell proliferation. The Women's Health Initiative study provided evidence for an increased risk of breast cancer in women treated with conjugated equine estrogens plus the synthetic progestin medroxyprogesterone acetate (MPA), compared with conjugated equine estrogens-only treatment. These findings continue to be discussed, and it remains to be clarified whether the results obtained for MPA in the Women's Health Initiative study are directly applicable to other progestins used in hormone therapy. In this study we compared in a mouse model the effects of the synthetic progestins, MPA, and drospirenone in two major target organs: the uterus and mammary gland. As quantitative measures of progestin activity, we analyzed maintenance of pregnancy, ductal side branching in the mammary gland, and proliferation of mammary and uterine epithelial cells as well as target gene induction in both organs. The outcome of this study is that not all synthetic progestins exhibit the same effects. MPA demonstrated uterine activity and mitogenic activity in the mammary gland at the same doses. In contrast, drospirenone behaved similarly to the natural hormone, progesterone, and exhibited uterine activity at doses lower than those leading to considerable proliferative effects in the mammary gland. We hypothesize that the safety of combined hormone therapy in postmenopausal women may be associated with a dissociation between the uterine and mammary gland activities of the progestin component. PMID- 18420742 TI - Estradiol-induced synaptic remodeling of tyrosine hydroxylase immunopositive neurons in the rat arcuate nucleus. AB - Gonadal steroids induce synaptic plasticity in several areas of the adult nervous system. In the arcuate nucleus of adult female rats, 17beta-estradiol triggers synaptic remodeling, resulting in a decrease in the number of inhibitory synaptic inputs, an increase in the number of excitatory synapses, and an enhancement of the frequency of neuronal firing. In the present paper, we studied the specificity of hormonal effects by determining the changes in synaptic connectivity of tyrosine hydroxylase (TH) immunoreactive (IR) neurons in the arcuate nucleus. We combined pre-embedding TH and post-embedding gamma aminobutyric acid (GABA) immunostaining, and performed unbiased stereological measurements in gonadectomized and 17beta-estradiol-treated rats. We conclude that the synaptic connectivity of the TH-IR neurons is different from the other, nonlabeled population, and the response to estradiol is not uniform. TH-IR (dopaminergic) arcuate neurons of both male and female rats have more GABAergic (inhibitory) axosomatic inputs than the nondopaminergic population. Our study shows that the effect of 17beta-estradiol is sex and cell specific in the sense that not all arcuate neurons are affected by the structural synaptic remodeling. In ovariectomized females hormone treatment decreased the numerical density of GABAergic axosomatic synapses on TH-IR, but not on nondopaminergic, neurons, whereas in orchidectomized males, 17beta-estradiol treatment increased inhibitory synapses onto nondopaminergic neurons but did not affect the number of inhibitory terminals onto TH-IR neurons. The hormone-induced plastic changes in synaptic connectivity of TH-IR neurons may serve as the morphological basis for the cyclical regulation of the anterior pituitary. PMID- 18420743 TI - Adrenal hypersensitivity precedes chronic hypercorticism in streptozotocin induced diabetes mice. AB - Previous studies have demonstrated that type 1 diabetes is characterized by hypercorticism and lack of periodicity in adrenal hormone secretion. In the present study, we tested the hypothesis that hypercorticism is initiated by an enhanced release of ACTH leading subsequently to adrenocortical growth and increased output of adrenocortical hormones. To test this hypothesis, we used the streptozotocin (STZ)-induced diabetes mouse model and measured hypothalamic pituitary-adrenal axis activity at different time points. The results showed that the expected rise in blood glucose levels induced by STZ treatment preceded the surge in corticosterone secretion, which took place 1 d after diabetes onset. Surprisingly, circulating ACTH levels were not increased and even below control levels until 1 d after diabetes onset and remained low until d 11 during hypercorticism. In response to ACTH (but not vasopressin), cultures of adrenal gland cells from 11-d diabetic mice secreted higher amounts of corticosterone than control cells. Real-time quantitative PCR revealed increased expression of melanocortin 2 and melanocortin 5 receptors in the adrenal glands at 2 and 11 d of STZ-induced diabetes. AVP mRNA expression in the paraventricular nucleus of the hypothalamus was increased, whereas hippocampal MR mRNA was decreased in 11-d diabetic animals. GR and CRH mRNAs remained unchanged in hippocampus and paraventricular nucleus of diabetic mice at all time points studied. These results suggest that sensitization of the adrenal glands to ACTH rather than an increase in circulating ACTH level is the primary event leading to hypercorticism in the STZ-induced diabetes mouse model. PMID- 18420745 TI - Oxidative stress regulates type 3 deiodinase and type 2 deiodinase in cultured rat astrocytes. AB - Type 2 deiodinase (D2) and type 3 deiodinase (D3) locally achieve the determination of the concentration of T3, which binds to the thyroid hormone receptor with high affinity. D2 converts T4 into T3, and D3 degrades T4 and T3. Neurons take up T3 released by astrocytes, the main cerebral site for the D2 expression. Because oxidative stress is believed to be involved in several neurological disorders, we explored the effects of oxidative stress on D3 and D2 in primary culture of rat astrocytes. H2O2 (250 microm) increased D3 activity with maximal effects around 8 h. Stimulation of D3 activity by H2O2 was synergistic with T4, phorbol ester, and also cAMP. H2O2 (250 microm) did not affect basal D2 activity but inhibited the stimulation of D2 activity by cAMP and factors implicating cAMP-independent pathways in astrocytes, TSH, and phorbol ester. N-Acetyl cysteine and selenium repletion, which respectively increase intracellular glutathione and glutathione peroxidase, inhibited D2 and D3 regulation by H2O2, whereas L-buthionine sulfoximine, which decreases intracellular glutathione, mimicked H2O2 effects. Oxidative stress up-regulated D3 and inhibited cAMP-stimulated D2 by transcriptional mechanisms. A decrease in cAMP by oxidative stress could contribute to the inhibition of cAMP-stimulated D2. Using specific inhibitors of signaling pathways, we show that the ERK pathway was required in D2 and D3 regulation by oxidative stress and that the p38 MAPK pathway was implicated in H2O2-induced D3. We suggest that the expected decrease in T3 might modulate the cellular injury of oxidative stress in some pathological brain conditions. PMID- 18420744 TI - Estrogen signaling characteristics of Atlantic croaker G protein-coupled receptor 30 (GPR30) and evidence it is involved in maintenance of oocyte meiotic arrest. AB - Human G protein-coupled receptor 30 (GPR30) mediates estradiol-17beta (E2) activation of adenylyl cyclase in breast cancer cells and displays E2 binding typical of membrane estrogen receptors (mERs). We identified a mER in Atlantic croaker ovaries with characteristics similar to those of human GPR30. To confirm the proposed role of GPR30 as a mER in this distantly related vertebrate group, we cloned GPR30 from croaker ovaries and examined its distribution, steroid binding, and signaling characteristics. Western blot analysis showed the GPR30 protein (approximately 40 kDa) is expressed on the plasma membranes of croaker oocytes and HEK293 cells stably transfected with GPR30 cDNA. Plasma membranes prepared from croaker GPR30-transfected cells displayed high-affinity, limited capacity, and displaceable binding specific for estrogens, characteristic of mERs. Consistent with previous findings with human GPR30, estrogen treatment of plasma membranes from both croaker ovaries and GPR30-transfected cells caused activation of a stimulatory G protein (Gs) resulting in increased cAMP production. Treatment with E2 as well as G-1, a specific GPR30 ligand, significantly reduced both spontaneous and progestin-induced maturation of both croaker and zebrafish oocytes in vitro, suggesting a possible involvement of GPR30 in maintaining oocyte meiotic arrest in these species. Injection of antisense oligonucleotides to GPR30 into zebrafish oocytes blocked the inhibitory effects of estrogen on oocyte maturation, confirming a role for GPR30 in the control of meiotic arrest. These findings further support our previous suggestion that GPR30 is a vertebrate mER. In addition, the results suggest GRP30 may play a critical role in regulating reentry into the meiotic cell cycle in fish oocytes. PMID- 18420746 TI - The Attitudes Toward Prostitution Scale: preliminary report on its development and use. AB - The Attitudes Toward Prostitution Scale (ATPS) assesses the beliefs of men who purchase sex from prostitutes. Items for the ATPS were administered to a U.S. multisite sample of 1,001 men who had been arrested for soliciting an undercover police officer and subsequently referred to a psychoeducational program. Principal axis factor analysis was conducted, and it yielded three valid factors: Social/Legal Support of Prostitution, Beliefs About Prostitutes, and Family Values Related to Prostitution. Scale-item reliability was assessed with Cronbach's alpha on a second independent sample of 74 men, which offered data to support ATPS internal consistency and reliability. Based on these results, scale scores were standardized for practical use in research and clinical applications, and interpretation norms were established for a population of men who buy sex. A brief case example illustrates the practical use of the ATPS for assessment and intervention planning for customers of sex workers. PMID- 18420747 TI - The effect of comorbidities upon ocular and systemic health-related quality of life. AB - AIM: The purpose of the study was to assess whether, and to what degree, comorbidities affect patient quality of life. METHODS: A cross-sectional, quality of-life study of 170 consecutive vitreoretinal patients compared the utility associated with a participant's primary (most incapacitating) disease and the utility associated with a grouping of all of the participants' diseases. The ocular diseases present included diabetic retinopathy (44%), macular degeneration (30%), lattice degeneration/retinal tear (14%), retinal vascular obstruction (5%), uveitis, macular oedema, macular pucker (5%) and others (2%). Participants underwent interviewer-administered, time trade-off utility questions for each disease, then for a compilation of all diseases. Their primary disease was defined by the lowest utility reported for a single disease, while other health conditions were considered comorbidities. A two-tailed, paired t test was used to compare the means of the primary disease utilities and compilation utilities. The study was powered to have a 90% chance of detecting an 8% difference in mean utility between the two utility groups RESULTS: The mean lowest utility for the most disabling single health condition (primary disease) was 0.82 (SD 0.22; 95% CI 0.79 to 0.85. The mean utility for the grouping together of all diseases was 0.80 (SD 0.24, 95% CI 0.76 to 0.84). No significant difference was found between the mean utilities of the two groups (p = 0.56). CONCLUSIONS: The overall health related quality of life of a patient in an ophthalmic population with serious diseases appears to be primarily determined by the single disease that most adversely affects the individual's quality of life. This conclusion has significant implications in clinical care and when considering the use of comorbidities in cost-utility analyses. PMID- 18420748 TI - Comparison of 4 mg versus 20 mg intravitreal triamcinolone acetonide injections. AB - AIMS: To compare the non-decanted (standard) 4 mg versus the decanted 20 mg intravitreal triamcinolone acetonide (IVTA) injections and to assess their effect on intraocular pressure (IOP). METHODS: We retrospectively reviewed the records of 92 consecutive eyes, which received an intravitreal injection of either dose of triamcinolone acetonide, at a single retina centre. The change in IOP (elevation of at least 5 mm Hg from baseline or above 21 mm Hg) was analysed with a multivariate logistic analysis. The mean follow-up period in both groups was 27 weeks. A subgroup analysis comparing vitrectomised to non-vitrectomised eyes in both groups was also performed. RESULTS: Of the 92 eyes, 46% (23 of 51) in the 4 mg group versus 30% (12 of 41) in the 20 mg group had an IOP >21 mm of Hg (p = 0.14) after a mean follow-up period of 27 weeks. The vitrectomised eyes (3 of 24) in the 20 mg group had a significantly lower rate of IVTA induced IOP elevation than the non-vitrectomised eyes (9 of 17) (p = 0.013). The IOP elevation occurred significantly earlier in the 4 mg group (vitrectomised eyes 27 (SD 43) days and non-vitrectomised eyes 61 (52) days) than in the 20 mg group (vitrectomised eyes 104 (56) days and non-vitrectomised eyes 119 (82) days), independent of the vitreous status (vitrectomised p = 0.05 and non-vitrectomised p = 0.04). The mean value of initial high IOP in the non-vitrectomised eyes was higher in the 4 mg group than in the corresponding 20 mg group (p = 0.048). CONCLUSION: Decanted 20 mg IVTA may not pose a significantly greater risk of IOP elevation than the 4 mg non-decanted IVTA. PMID- 18420749 TI - Intravitreal triamcinolone versus laser photocoagulation for persistent diabetic macular oedema. AB - AIM: To determine if repeated intravitreal triamcinolone improves best corrected visual acuity at 1 year compared with conventional laser therapy for persistent diabetic macular oedema. METHODS: 88 eyes with persistent clinically significant macular oedema, after at least one prior laser photocoagulation, were included in this prospective randomised controlled trial. 43 patients were randomised to 4 mg of intravitreal triamcinolone (TA) and 45 to laser photocoagulation. The primary endpoint was the proportion of patients who improved by 15 Early Treatment of Diabetic Retinopathy Study (ETDRS) letters at 12 months in TA versus laser groups. Secondary endpoints were the change in mean best corrected visual acuity, difference in macular thickness and macular volume and adverse event reporting in particular elevated intraocular pressure at 12 months. RESULTS: Improvement in > or =15 ETDRS letters occurred in two of 42 patients in the TA group (4.8%) and in five of 41 (12.2%) patients in the laser group (p = 0.265). At baseline, the mean ETDRS scores at baseline were 54.4 letters in the TA group and 53.0 letters in the laser group. At 12 months, these were 54.5 and 54.6, respectively. Optical coherence tomography showed a reduction in central macular thickness of 82.0 microm with TA and 62.3 microm with laser at 12 months. There was one case of sterile endophthalmitis. 22 out of 43 patients in the triamcinolone group required ocular antihypertensives. CONCLUSION: This study did not show a benefit from intravitreal triamcinolone over conventional laser therapy for patients with chronic diabetic macular oedema. PMID- 18420750 TI - Sources of patient knowledge and financing of cataract surgery in rural China: the Sanrao Study of Cataract Outcomes and Up-Take of Services (SCOUTS), report 6. AB - AIM: To study patient sources of knowledge about cataract surgical services, and strategies for financing surgery in rural China. DESIGN: Cross-sectional case series. METHODS: Patients undergoing cataract surgery by local surgeons in a government, village-level facility in Sanrao, Guangdong between 8 August and 31 December 2005 were examined and had standardised interviews an average of 12 months after surgery. RESULTS: Of 313 eligible patients, 239 (76%) completed the questionnaire. Subjects had a mean (SD) age of 69.9 (10.2) years, 36.4% (87/239) were male, and 87.0% (208/239) had been blind (presenting visual acuity < or = 6/60) before surgery. Word-of-mouth advertising was particularly important: 198 (85.0%) of the subjects knew a person who had undergone cataract surgery, of whom 191 (96.5%) had had cataract surgery at Sanrao itself. Over 70% of subjects (166/239) watched TV daily, whereas 80.0% (188/239) "never" read the newspaper. Nearly two-thirds of suggestions from participants (n = 211, 59.6%) favoured either TV advertisements or word-of-mouth to publicise the programme. While the son or daughter had paid for surgery in over 70% of cases (164/233), the patient's having paid without help was the sole predictor of undergoing second eye surgery (OR 2.27 (95% CI 1.01 to 5.0, p = 0.04)). DISCUSSION: Strategies to increase uptake of cataract surgery in rural China may benefit from enhancing word-of-mouth advertising (such as with pseudophakic motivators), using television advertising where affordable, and micro-credit or other programmes to enable patients to pay their own fees, thus increasing uptake of second-eye surgery. PMID- 18420751 TI - Performance of the compensation comparison method for retinal straylight measurement: effect of patient's age on repeatability. AB - AIM: The assessment of repeatability and reproducibility of retinal straylight measurements with the C-Quant straylight meter (Oculus AG, Germany) and the effect of patient's age on the instrument performance are tested with a series of experiments. METHODS: First, 20 eyes from 20 subjects (mean age 26.9 (SD 2.7) years, mean refractive error -1.34 (2.72) D) were examined with the C-Quant straylightmeter, taking 10 consecutive readings. Five subjects were also examined on five consecutive days to assess reproducibility. Additionally, repeated measures of straylight from 84 subjects of ages ranging from 19 to 86 years (mean (SD): 42.4 (24.0) years) were retrospectively analysed to assess the effect of patient's age on repeatability. RESULTS: The results failed to show significant differences between the readings taken within the same session (mean (0.07), p>0.05) or between sessions (mean (0.05), p>0.05). Variability of intrasession measurements was not significant for subjects of different age (p = 0.094). CONCLUSION: It may be concluded that the C-Quant straylightmeter is repeatable and reliable for the assessment of retinal straylight in human eyes. Age of the patient does not decrease repeatability, even though they feel more insecure about their ability to perform the test. PMID- 18420752 TI - Physical exercise and psychological well-being among people with chronic illness and disability: a grounded approach. AB - This qualitative study describes the effect of exercise on psychological well being among individuals with chronic illnesses and disabilities such as stroke, cancer, diabetes and arthritis. Eleven users of a physical disability daycentre completed a novel, six-month, group-based programme of seated exercise provided by the first author. Results from grounded analyses of in-depth interviews provide evidence that exercise offers a powerful means to actively manage mood problems associated with disability, to preserve and cultivate personal identity in a positive manner and to connect with others and one's own body in ways which limit the development of illness-related psychological distress. Implications for the development of further research and future provision of similar services are discussed. PMID- 18420753 TI - Gender differences in smoking and meanings of smoking in Asian-American college students. AB - This study examined the meanings of smoking among Asian-America college students and the associations between those meanings and their smoking behaviors. Data were collected from two postsecondary institutions in southern California. A total of 460 students with 205 males and 255 females completed the survey. The odds ratio of each meaning to students' smoking behaviors was calculated. Spearman's correlational analyses were run to make comparisons between genders. Males who endorsed social meanings were more likely to smoke while females who endorsed personal meanings were more likely to smoke. Gender-tailored smoking prevention programs may be needed for Asian-American college students. PMID- 18420754 TI - The affective impact of exercise intensity that slightly exceeds the preferred level: 'pain' for no additional 'gain'. AB - We examined the affective consequences of an exercise intensity that slightly exceeded the preferred level. Twenty-five middle-age sedentary women participated in two 20-min treadmill exercise bouts, one during which they could select the speed and one during which the speed was 10 per cent higher than the self selected. During the bout at self-selected intensity, ratings of pleasure remained stable, whereas during the imposed-intensity bout pleasure decreased. Therefore, even a minor increase in exercise intensity beyond the level that a new exerciser would have self-selected can bring about a decrease in pleasure. Over time, such experiences could lower adherence. PMID- 18420755 TI - Sequestering of suffering: critical discourse analysis of natural disaster media coverage. AB - This article is a critical discourse analysis of the local print-news media coverage of the recovery process in two rural communities following a devastating forest fire. Two hundred and fifty fire-related articles from the North Thompson Star Journal (2003) were analyzed. Results revealed a neoliberal discursive framing of recovery, emphasizing the economic-material aspects of the process and a reliance on experts. A sequestering of suffering discourse promoted psychological functionalism and focused attention on a return to normalcy through the compartmentalization of distress. The dominant 'voice' was male, authoritative, and institutionalized. Implications for disaster recovery and potential health consequences are discussed. PMID- 18420756 TI - Psychological and behavioural impact of genetic testing smokers for lung cancer risk: a phase II exploratory trial. AB - The behavioural and psychological impact of genetic testing for lung cancer susceptibility was examined among smokers (N = 61) who were randomly allocated to a GSTM1 genetic testing group (with GSTM1-missing or GSTM1-present result) or no test control group. The GSTM1-missing (higher risk) group reported greater motivation to quit smoking, and both genetic testing groups reported lower depression than the control group at one-week follow-up (p < .05 for all). Differences were not significant at two months follow-up. This study indicates the feasibility of much-needed research into the risks and benefits for individuals of emerging lifestyle-related genetic susceptibility tests. PMID- 18420757 TI - The role of exercise dependence for the relationship between exercise behavior and eating pathology: mediator or moderator? AB - Our study examined the potential mediating or moderating effect of exercise dependence on the exercise-eating pathology relationship. Female university students (N = 330) completed Internet-based self-report measures of exercise behavior, exercise dependence, and eating pathology. Exercise dependence served as a mediator for the relationship between exercise and eating pathology. This unidirectional causal model suggests that an individual's pathological motivation or compulsion to exercise is the critical mediating component in the exercise eating pathology relationship. The best target for removing the link between exercise behavior and eating pathology may be reformulating exercise dependence symptoms. PMID- 18420758 TI - How well do women adapt to changes in their body size and shape across the course of pregnancy? AB - This study examined body image across pregnancy. Pregnant women ( N = 158) completed measures of general attractiveness, feeling fat, fitness and strength, salience of weight and shape, and ideal and current body size at pre-pregnancy (retrospective), and in early, middle and late pregnancy. Body image was found to be fairly stable across pregnancy such that women who started with greater body concerns maintained them over time. Although women were least satisfied with their stomach size at late pregnancy, women's ideal body shape increased in parallel with increases in body size. Women with the most body concerns reported more depressive symptoms, tendency towards dieting, and smoking during pregnancy suggesting they were at greater risk in terms of health and well-being during pregnancy. PMID- 18420759 TI - Dispositional predictors of complementary medicine and vitamin use in students. AB - The Rational-Experiential Inventory (REI) and Tellegen Absorption questionnaire were completed by 131 undergraduate students who also completed a scale measuring complementary medicine use (with separate entries for practitioner and self delivered use) and vitamin/mineral use. Less rationality and greater absorption predicted greater use of practitioner-delivered and self-delivered complementary medicine, and vitamin/mineral use. Logistic regression showed that rationality and absorption were independent predictors of these behaviours. In addition to confirming previous research, this is the first demonstration that complementary medicine and vitamin/mineral use are predicted by the same variables, including less rationality. PMID- 18420760 TI - The effects of unmitigated communion and life events among women with fibromyalgia syndrome. AB - This study examined the effects of the personality trait of unmitigated communion (UC) within the context of positive and negative relationship events. Ninety-one women diagnosed with fibromyalgia syndrome completed initial measures of UC as well as weekly measures of positive and negative relationship events and positive and negative affect. Positive events were related to positive affect while negative events were related to negative affect. Negative and positive affect were also predicted from the interaction of UC and negative events. These results suggest the interrelationships among UC and relationship events can combine in meaningful ways to predict psychological well-being. PMID- 18420761 TI - Chronic fatigue syndrome in male Gulf war veterans and civilians: a further test of the single syndrome hypothesis. AB - Different modes of fatigue onset in male Gulf War veterans versus male civilians raise the possibility that chronic fatigue syndrome (CFS) may not be a single disease entity. We addressed this issue by comparing 45 male veterans with CFS to 84 male civilians who satisfied identical case criteria. All were evaluated for fibromyalgia (FM), multiple chemical sensitivity and psychiatric comorbidity. CFS was more likely to present in a sudden flu-like manner in civilians than veterans (p < .01) and comorbid FM was more prevalent in civilians (p < .01). These findings question the assumption that all patients with CFS suffer from the same underlying disorder. PMID- 18420762 TI - Emotions and their cognitive precursors: responses to spontaneously identified stressful events among hospital nurses. AB - This study used Smith and Lazarus's appraisal model to explore nurses' emotional reactions to self-identified distressing work situations. Key situations reported typically involved time or interpersonal pressure, coping with errors at work, difficulties with patients, and managerial conflict. The strongest emotions experienced were anger and frustration. Anxiety and sadness were less endorsed. Most nurses felt able to cope emotionally and problem-solve solutions to the problems they faced. The findings were generally supportive of Smith and Lazarus's theory, with the exception of the emotion of sadness. The implications of the results are discussed. PMID- 18420763 TI - Symptoms of depression and anxiety among a sample of South African patients living with a chronic illness. AB - The present study reports on a survey conducted among 119 patients receiving treatment for diabetes or hypertension at semi-rural community health care clinics in South Africa. Participants completed the Hopkins Symptom Checklist (HSCL) and the Beck Depression Inventory (BDI). On the HSCL, 38.5 per cent of the sample scored in the elevated range. On the BDI, 19.8 per cent of the sample fell in or above the moderate range for depression. The results suggest that a considerable proportion of the sample may be experiencing significant psychiatric difficulty, for which they may not be receiving treatment. PMID- 18420764 TI - Postpartum smoking cessation and relapse prevention intervention: a structural equation modeling application to behavioral and non-behavioral outcomes of a randomized controlled trial. AB - The aim of the study was to test the effectiveness of a postpartum smoking cessation and relapse prevention intervention. Structural equation modeling techniques were applied to evaluate the impact of the intervention on smoking behavior and on non-behavioral variables derived from the Transtheoretical Model (TTM). Women were randomized to an intervention (I) and control group (C). Smoking status, TTM-variables, and control variables were assessed four weeks, six and 12 months postpartum. Membership in the intervention group significantly predicted non-smoking and higher self-efficacy six months, but not one year postpartum, after controlling for demographic, smoking, and postpartum risk variables. PMID- 18420765 TI - The impact of breast scarring on perceptions of attractiveness: an experimental study. AB - This study explored whether breast scarring following a lumpectomy generated negative judgements using an experimental between subjects factorial design. Eight images of women varying in terms of breast scarring (present vs. absent), beauty (average vs. high) and celebrity (as a proxy for familiarity; present vs. absent) were rated by 232 participants. The results showed that scarring resulted in lower ratings of attractiveness and self esteem and higher ratings of femininity. Scarred celebrities were judged as particularly unattractive. Women's assumption that they will be judged more harshly following breast cancer surgery maybe reflected in the actual ratings of others. PMID- 18420766 TI - What do coronary artery disease patients think about their treatments? An assessment of patients' treatment representations. AB - This article investigates patients' beliefs about the intervention offered to manage their illness. Coronary artery disease (CAD) patients, 70 of whom were undergoing medication, 71 to undergo angioplasty and 73 to undergo surgery, completed a 58-item questionnaire regarding their treatment beliefs. Responses were subject to principal components analysis, which indicated four factors accounting for 36.7 per cent of the variance. After excluding extraneous items, the final questionnaire consisted of 27 items, clustered around four components: treatment-value, treatment-concerns, decision-satisfaction and cure. A coherent set of subscale inter-correlations and ANCOVAs examining treatment group differences on these sub-scales showed a logical, explicable pattern of group differences reflecting the distinctive natures of each treatment and demonstrated discriminant validity. Correlations with other scales provided evidence of construct validity. PMID- 18420767 TI - Development and validation of a religious health fatalism measure for the African American faith community. AB - Health researchers struggle to understand barriers to improving health in the African-American community. The African-American church is one of the most promising venues for health promotion, disease prevention, and disparities reduction. Religious fatalism, the belief that health outcomes are inevitable and/or determined by God, may inhibit healthy behaviors for a subset of religious persons. This study reports the development and validation of the Religious Health Fatalism Questionnaire, a measurement tool for studying faith-related health beliefs in African-Americans. Participants included 276 members of seven predominantly African-American churches. Factor analysis indicated three dimensions: (1) Divine Provision; (2) Destined Plan; and (3) Helpless Inevitability. Evidence is presented for the reliability, convergent and predictive validity of the Religious Health Fatalism Questionnaire. PMID- 18420768 TI - The relationship between negative communication and body image dissatisfaction in adolescent females with Type 1 Diabetes Mellitus. AB - Maladaptive eating attitudes and behaviors are common in adolescent females with Type 1 Diabetes Mellitus (T1DM). This research assessed potential pathways through which individual and familial factors relate to treatment adherence and glycemic control. Seventy-five females with T1DM (aged 11-17 years) and their mothers completed questionnaires regarding communication, diabetes management, and eating attitudes and behaviors. Hierarchical regression analyses found that body image dissatisfaction moderates the relationship between negative communication and maladaptive eating attitudes and behaviors. Treatment adherence mediates the relationship between maladaptive eating attitudes and behaviors and glycemic control. None of the participants endorsed omitting insulin for the purposes of weight management. This study highlights the need to address familial and individual factors along with treatment adherence within this population. PMID- 18420769 TI - Ways of coping and perceived stress in women with spinal cord injury. AB - Using a cross-sectional design, this research aimed to assess whether a three factor model of Positive Reappraisal, Escape-Avoidance, and Seeking Social Support based on the Ways of Coping Questionnaire (WOCQ) appropriately depicts coping within a sample of women with spinal cord injury (SCI). Forty-four community-dwelling women with spinal cord injury were interviewed from two urban rehabilitation facilities in the Midwestern United States. The main outcome measures used were the Perceived Stress Scale (PSS) and the WOCQ. The Positive Reappraisal, Escape-Avoidance, and Seeking Social Support scales of the WOCQ significantly accounted for variance in perceived stress. These three scales appear to be most relevant to perceived stress in women with SCI. Implications for coping research in this population are discussed. PMID- 18420770 TI - An alternative framework for understanding women's caregiving stress: a qualitative application of the ways of coping model. AB - In this article we present an alternative to inventory checklist approaches to measuring coping. We utilized a qualitative approach to explore themes of problem focused, emotion-focused and meaning-focused coping in a sample of 98 women from diverse backgrounds and caregiving situations. Their narratives reflected the ;goodness of fit' coping framework; that different strategies are more effective for stress reduction in relation to the amount of perceived control. Using a narrative approach allowed us to situate coping within the context of the caregiving experiences that the women discussed. This permits a broader understanding of how coping strategies can help women to navigate caregiving and reduce stress. PMID- 18420771 TI - Self-efficacy for exercise in cardiac rehabilitation: review and recommendations. AB - Despite the numerous physical and psychosocial benefits of exercise for coronary heart disease survivors, non-adherence to cardiac rehabilitation (CR) exercise is a major problem. Adherence to the lifestyle behavior change associated with CR involves both physical and self-regulatory skills. While self-regulatory efficacy is clearly linked to exercise adherence and adjustment, the literature on the relationship between self-efficacy and exercise among CR participants has not been systematically reviewed. A search of relevant databases identified 41 CR studies. Few studies measured self-regulatory efficacy for actions that facilitate adherence. Most studies examined self-efficacy during the intensive center-based phase of CR, with little attention to long-term maintenance. The CR literature could benefit by examining (a) self-efficacy as a major rehabilitation outcome, (b) measurement of self-regulatory efficacy for behavior change, (c) suspected moderators of self-efficacy (i.e. gender, age), and (d) self-efficacy relative to maintenance. PMID- 18420772 TI - Patient satisfaction and sustained outcomes of drug abuse treatment. AB - The authors investigated the relationship between patients' self-rated satisfaction with treatment services during and shortly after treatment with their drug use outcomes at one year follow-up, using a US national panel survey of patients in 62 methadone, outpatient, short-term residential, and long-term residential programs. A favorable evaluation of treatment near the time of discharge had a significant positive relationship with drug use improvement outcomes approximately one year later, independent of the separately measured effects of treatment duration, counseling intensity, patient adherence to treatment protocols, pre-treatment drug use patterns, and other characteristics of patients and treatment programs. PMID- 18420773 TI - Understanding hope in the face of an HIV diagnosis and high-risk behaviors. AB - The purpose of this article is to: (a) explore and describe experiences of hope in the context of high-risk behaviors; and (b) identify sources of hope for participants dealing with an HIV diagnosis. Following qualitative case study methodology along with philosophical hermeneutics, 12 people diagnosed with HIV or AIDS were interviewed to understand their experiences of hope. Participants were selected based on self-report for continuing or increasing high-risk behaviors following diagnosis. Results include participant descriptions of hope as well as five higher order categories that highlight sources of hope following an HIV diagnosis. Implications for treatment of those newly diagnosed and engaged in high-risk behaviors are discussed. PMID- 18420774 TI - Correlates of self-efficacy among rural smokers. AB - Self-efficacy has been related to intent to stop smoking, abstinence success, and risk for relapse. Because limited research exists regarding self-efficacy among rural smokers, the current study examined correlates of self-efficacy among rural primary care patients smoking > or =10 cigarettes per day. Participants completed a telephone survey assessing demographics, smoking history, and psychosocial variables (e.g. motivation, depression). Among the 750 participants, lower self efficacy was correlated with high depression scores, shorter previous abstinence, lower autonomous motivation, younger age, higher nicotine dependence, readiness to quit, and being female. Future studies should examine the potential to improve self-efficacy by addressing depression and autonomous motivation. PMID- 18420775 TI - Romantic relationships and eating regulation: an investigation of partners' attempts to control each others' eating behaviors. AB - The current study examines eating regulation in the context of romantic relationships. One hundred and four heterosexual couples provided information about their weight, satisfaction with their partners' bodies, weight concerns, healthy dieting behaviors, and attempts to regulate their partners' eating behaviors. Results indicated that women were more likely to monitor their partners' eating behaviors than were men. Both men and women who attempted to regulate their partners' eating behaviors tended to have relatively heavy partners and tended to be dissatisfied with their partners' bodies. However, only women's attempts to regulate men's eating behaviors were associated with men's weight concerns and healthy dieting behaviors. PMID- 18420776 TI - The effects of idealized fitness images on anxiety, depression and global mood states in college age males and females. AB - The primary purpose of this investigation was to quantify the effects of fitness media exposure on psychological affect utilizing the Solomon 4-group design. Volunteers were counterbalanced and randomly assigned to 30 minutes of viewing either a fitness or control magazine. State anxiety (STAI), tension, depression, anger vigor, fatigue, confusion, and global mood (POMS) were measured. Results indicate that pre-test sensitization was not a significant factor when testing the psychological factors in this investigation. Additionally, the outcomes associated with viewing ultra-fit images in popular media are not a benign experience and merit further examination. PMID- 18420777 TI - Defining the natural history of MS: the need for complete data and rigorous definitions. PMID- 18420778 TI - Cancer risk and impact of disease-modifying treatments in patients with multiple sclerosis. AB - BACKGROUND: Prior to the era of immunomodulating or immunosuppressive (IS) treatments Multiple Sclerosis (MS) was linked to reduced rates of cancer. Method A descriptive study of MS patients with a documented oncological event was performed. From 1 January 1995 to 30 June 2006, we collected and studied the profile of 7,418 MS patients gathered from nine French MS centers. We evaluated the incidence of cancer in a Cancer Risk In MS Cohort. RESULTS: Thirty one patients (1.75%) with confirmed MS had a history of cancer: mean age at MS diagnosis of 37.9 years and a mean age at cancer diagnosis of 46.4 years. The most frequent cancers were breast (34.5%), gynecological (12.5%), skin (10.2%), acute leukemia and lymphoma (5.9%), digestive (8.8%), kidney and bladder (5.1%), lung (3.4%) and central nervous system (3%). Calculated standardized incidence rates were 0.29 (0.17-0.45) for men and 0.53 (0.42-0.66) for women. The incidence of cancer in this MS population was lower than that expected for the general population. Matched to age, gender and histology, cancers in MS were associated with a young age and exposure to IS treatments. When considering all patients, treated patients had a 3-fold higher risk of developing cancer, if they had a history of IS (P = 0.0035). For treated patients, the cancer sites were more likely the breast, the urinary tract, the digestive system and the skin. CONCLUSION: Our data suggest that MS patients do not have an increased risk of cancer. Rather for several types of cancer a significantly reduced risk was observed, except for breast cancer in women treated with IS. The relative increased risk of breast cancer in MS women under IS treatment warrants further attention. PMID- 18420779 TI - Comparative analysis of CYP3A heteroactivation by steroid hormones and flavonoids in different in vitro systems and potential in vivo implications. AB - A systematic analysis of the heteroactivation of CYP3A-mediated carbamazepine 10,11-epoxidation has been investigated in three different in vitro systems, namely recombinant CYP3A4 and CYP3A5, human liver microsomes (HLMs) and cryopreserved human hepatocytes. The effect of 10 endogenous steroids and flavonoids was studied over a range of substrate and effector concentrations. A novel heteroactivation model was used to obtain the parameters EC(200) (concentration of effector required to produce 200% control) and heteroactivation ratio (the ratio of maximum observed reaction velocity to control). The EC(200) values obtained in HLMs and human hepatocytes were corrected for nonspecific binding. Heteroactivation of CYP3A5 has been demonstrated with mean heteroactivation ratios in CYP3A5, HLMs and hepatocytes on average 2-fold greater than in recombinant CYP3A4 for most of the effectors investigated. In recombinant CYP3A4, heteroactivation was greatest at substrate concentrations below K(m). Heteroactivation increased with effector concentration in a nonlinear manner and differed between effectors (mean heteroactivation ratios varied up to 12-fold). A greater extent of heteroactivation was observed in HLMs than in human hepatocytes for steroid effectors, but the opposite was true for flavonoid effectors. The observed heteroactivation of CYP3A in intact cells supports an in vivo relevance. From the in vitro heteroactivation data, a significant increase in clearance in vivo was predicted for substrates with a high dependence on CYP3A4 to the overall elimination, indicating that heteroactivation of CYP3A may be a potential source of interindividual variability. PMID- 18420780 TI - Functional expression and comparative characterization of nine murine cytochromes P450 by fluorescent inhibition screening. AB - The increasing number of transgenic or gene knockout mouse models generated for use in drug metabolism studies has meant that a greater understanding of the function and substrate specificities of murine cytochromes P450 (P450s) has become essential, particularly with the recent advances in "humanized" mouse models. In this study, we have heterologously expressed nine murine P450s- Cyp1a1, Cyp1a2, Cyp1b1, Cyp2a4, Cyp2b20, Cyp2c29, Cyp2d22, Cyp2e1, and Cyp3a11- individually with human P450 oxidoreductase to generate functional monooxygenase systems in Escherichia coli. We have identified a suitable fluorogenic probe for each P450 and determined the apparent kinetic parameters. These probes have enabled the screening of a panel of 31 test compounds classified as "drugs," "natural compounds," "endogenous compounds," and "pesticides" by measurement of IC(50), thus allowing the comparison of binding affinities. Human P450s CYP2C9, CYP2D6, and CYP3A4 were also included in the study to enable direct comparisons to be made with the mouse enzymes. Although there were general similarities between human and mouse P450s, perhaps the most significant finding in this study was the observation that, despite 77% amino acid identity, Cyp2d22 and CYP2D6 were remarkably dissimilar in a range of enzymatic properties, with potentially serious implications for pharmacokinetic studies using CYP2D substrates. The data presented in this study provide a solid foundation with which to assess the degree of similarity (or difference) between mouse and human P450s involved in xenobiotic metabolism and can be used as a basis for further studies. PMID- 18420781 TI - An in vitro mechanistic study to elucidate the desipramine/bupropion clinical drug-drug interaction. AB - There are documented clinical drug-drug interactions between bupropion and the CYP2D6-metabolized drug desipramine resulting in marked (5-fold) increases in desipramine exposure. This finding was unexpected as CYP2D6 does not play a significant role in bupropion clearance, and bupropion and its major active metabolite, hydroxybupropion, are not strong CYP2D6 inhibitors in vitro. The aims of this study were to investigate whether bupropion's reductive metabolites, threohydrobupropion and erythrohydrobupropion, contribute to the drug interaction with desipramine. In human liver microsomes using the CYP2D6 probe substrate bufuralol, erythrohydrobupropion and threohydrobupropion were more potent inhibitors of CYP2D6 activity (K(i) = 1.7 and 5.4 microM, respectively) than hydroxybupropion (K(i) = 13 microM) or bupropion (K(i) = 21 microM). Furthermore, neither bupropion nor its metabolites were metabolism-dependent CYP2D6 inhibitors. Using the in vitro kinetic constants and estimated liver concentrations of bupropion and its metabolites, modeling was able to predict within 2-fold the increase in desipramine exposure observed when coadministered with bupropion. This work indicates that the reductive metabolites of bupropion are potent competitive CYP2D6 inhibitors in vivo and provides a mechanistic explanation for the clinical drug-drug interaction between bupropion and desipramine. PMID- 18420782 TI - Use of online microdialysis sampling to determine the in vivo rate of phenol glucuronidation in rainbow trout. AB - A quantitative microdialysis (MD) sampling method was used to study phenol (PH) glucuronidation in vivo in rainbow trout. The method employs internal calibrators to account for changes in MD probe performance (in vitro-to-in vivo and sample-to sample) and yields data of high temporal resolution that are well suited for developing kinetic models. Initially, trout were dosed with phenyl glucuronide (PG) by intravascular infusion for 24 h and then depurated for 48 h. Measured concentrations of PG in blood were well described by a one-compartment clearance volume model. Massbalance calculations showed that 93% of infused PG was eliminated in urine during the depuration period. Peak concentrations of PG in urine averaged 3.4 times higher than those in blood, and the fitted PG clearance constant (15.7 ml/kg/h) was about 2.6 times higher than the reported glomerular filtration rate for trout. These findings confirm earlier work suggesting that PG is actively secreted by the trout kidney. In a second set of experiments, trout were exposed continuously to PH in water. In vivo rate constants for PH glucuronidation were estimated using a pair of linked (PH and PG) one-compartment clearance-volume models. Expressed on a whole-fish basis, the glucuronidation rate averaged 0.049/h, which was about 7% of the total rate of PH elimination. This study demonstrates the utility of quantitative MD sampling for kinetic studies of xenobiotic metabolism in fish. PMID- 18420783 TI - Cellular accumulation of cholyl-glycylamido-fluorescein in sandwich-cultured rat hepatocytes: kinetic characterization, transport mechanisms, and effect of human immunodeficiency virus protease inhibitors. AB - The present study was aimed at characterizing the in vitro cellular uptake mechanism and kinetics of the bile salt analog cholylglycylamido-fluorescein (CGamF) in sandwich-cultured rat hepatocytes (SCRHs). Concentration-dependent inhibition of active CGamF accumulation by seven human immunodeficiency virus (HIV) protease inhibitors (PIs) was also determined and compared with inhibition data obtained with taurocholate (TC) as a substrate. A K(m) value of 9.3 +/- 2.6 microM was obtained for saturable CGamF accumulation in SCRHs. The organic anion transporting polypeptide (Oatp) inhibitor rifampicin (100 microM) inhibited CGamF (1 microM) accumulation in SCRHs by 72%; sodium depletion did not further reduce CGamF accumulation. In contrast, TC accumulation was reduced by only 25% in the presence of rifampicin, whereas additional sodium depletion resulted in a complete loss of TC accumulation. These data imply that Oatp(s) and sodium taurocholate-cotransporting polypeptide preferentially mediate hepatic uptake of CGamF and TC, respectively. Coincubation of CGamF with HIV PIs (amprenavir, atazanavir, darunavir, indinavir, nelfinavir, ritonavir, saquinavir) revealed that five of them had a concentration-dependent inhibitory effect on CGamF accumulation in SCRHs, with IC(50) values between 0.25 +/- 0.07 and 43 +/- 12 microM. The rank order for inhibition of CGamF accumulation in SCRHs was: ritonavir >> saquinavir > atazanavir > darunavir > amprenavir. Indinavir (up to 100 microM) did not alter CGamF accumulation, whereas nelfinavir solubility was limited to 10 microM. Taken together, these findings illustrate the utility of CGamF as a suitable probe (complementary to TC) for rapid in vitro determination of interaction potential with sodium-independent uptake mechanisms (likely Oatps) in rat liver. PMID- 18420784 TI - Metabolism and disposition of dasatinib after oral administration to humans. AB - SPRYCEL (dasatinib, BMS-354825; Bristol-Myers Squibb, Princeton, NJ), a multiple kinase inhibitor, is currently approved to treat chronic myelogenous leukemia and Philadelphia chromosome-positive acute lymphoblastic leukemia tumors in patients who are resistant or intolerant to imatinib mesylate (Gleevec; Novartis, Basel, Switzerland). After a 100-mg single p.o. dose of [(14)C]dasatinib to healthy volunteers, the radioactivity was rapidly absorbed (T(max) approximately 0.5 h). Both dasatinib and total radioactivity (TRA) plasma concentrations decreased rapidly with elimination half-life values of <4 h. Dasatinib was the major drug related component in human plasma. At 2 h, dasatinib accounted for 25% of the TRA in plasma, suggesting that metabolites contributed significantly to the total drug-related component. There were many circulating metabolites detected that included hydroxylated metabolites (M20 and M24), an N-dealkylated metabolite (M4), an N-oxide (M5), an acid metabolite (M6), glucuronide conjugates (M8a,b), and products of further metabolism of these primary metabolites. Most of the administered radioactivity was eliminated in the feces (85%). Urine recovery accounted for <4% of the dose. Dasatinib accounted for <1 and 19% of the dose in urine and feces, respectively, suggesting that dasatinib was well absorbed after p.o. administration and extensively metabolized before being eliminated from the body. The exposures of pharmacologically active metabolites M4, M5, M6, M20, and M24 in patients, along with their cell-based IC(50) for Src and Bcr-Abl kinase inhibition, suggested that these metabolites were not expected to contribute significantly toward in vivo activity. PMID- 18420785 TI - Biotransformation of [14C]dasatinib: in vitro studies in rat, monkey, and human and disposition after administration to rats and monkeys. AB - This study describes the in vitro metabolism of [(14)C]dasatinib in liver tissue incubations from rat, monkey, and human and the in vivo metabolism in rat and monkey. Across species, dasatinib underwent in vitro oxidative metabolism to form five primary oxidative metabolites. In addition to the primary metabolites, secondary metabolites formed from combinations of the oxidative pathways and conjugated metabolites of dasatinib and its oxidative metabolites were also observed in hepatocytes incubations. In in vivo studies in rats and monkeys, the majority of the radioactive dose was excreted in the bile and feces. In bile duct cannulated monkeys after an i.v. dose, 13.7% of the radioactive dose was excreted in the feces through direct secretion. Dasatinib comprised 56 and 26% of the area under the curve (AUC) (0-8 h) of total radioactivity (TRA) in plasma, whereas multiple metabolites accounted for the remaining 44 and 74% of the AUC (0-8 h) of TRA for rats and monkeys, respectively. In rat and monkey bile, dasatinib accounted for < 12% of the excreted dose, suggesting that dasatinib was extensively metabolized before elimination. The metabolic profiles in bile were similar to the hepatocyte profiles. In both species, a large portion of the radioactivity excreted in bile (> or = 29% of the dose) was attributed to N oxides and conjugated metabolites. In rat and monkey feces, only the oxidative metabolites and their further oxidation products were identified. The absence of conjugative or N-oxide metabolites in the feces suggests hydrolysis or reduction, respectively, in the gastrointestinal tract before elimination. PMID- 18420786 TI - Polyinosinic acid enhances delivery of adenovirus vectors in vivo by preventing sequestration in liver macrophages. AB - Adenovirus is among the preferred vectors for gene therapy because of its superior in vivo gene-transfer efficiency. However, upon systemic administration, adenovirus is preferentially sequestered by the liver, resulting in reduced adenovirus-mediated transgene expression in targeted tissues. In the liver, Kupffer cells are responsible for adenovirus degradation and contribute to the inflammatory response. As scavenger receptors present on Kupffer cells are responsible for the elimination of blood-borne pathogens, we investigated the possible implication of these receptors in the clearance of the adenovirus vector. Polyinosinic acid [poly(I)], a scavenger receptor A ligand, was analysed for its capability to inhibit adenovirus uptake specifically in macrophages. In in vitro studies, the addition of poly(I) before virus infection resulted in a specific inhibition of adenovirus-induced gene expression in a J774 macrophage cell line and in primary Kupffer cells. In in vivo experiments, pre administration of poly(I) caused a 10-fold transient increase in the number of adenovirus particles circulating in the blood. As a consequence, transgene expression levels measured in different tissues were enhanced (by 5- to 15-fold) compared with those in animals that did not receive poly(I). Finally, necrosis of Kupffer cells, which normally occurs as a consequence of systemic adenovirus administration, was prevented by the use of poly(I). No toxicity, as measured by liver-enzyme levels, was observed after poly(I) treatment. From our data, we conclude that poly(I) can prevent adenovirus sequestration by liver macrophages. These results imply that, by inhibiting adenovirus uptake by Kupffer cells, it is possible to reduce the dose of the viral vector to diminish the liver-toxicity effect and to improve the level of transgene expression in target tissues. In systemic gene-therapy applications, this will have great impact on the development of targeted adenoviral vectors. PMID- 18420787 TI - Restriction of human herpesvirus 6B replication by p53. AB - Human herpesvirus 6B (HHV-6B) induces significant accumulation of p53 in both the nucleus and cytoplasm during infection. Activation of p53 by DNA damage is known to induce either growth arrest or apoptosis; nevertheless, HHV-6B-infected cells are arrested in their cell cycle independently of p53, and only a minor fraction of the infected cells undergoes apoptosis. Using pifithrin-alpha, a p53 inhibitor, and p53-null cells, this study showed that infected epithelial cells accumulated viral transcripts and proteins to a significantly higher degree in the absence of active p53. Moreover, HHV-6B-induced cytopathic effects were greatly enhanced in the absence of p53. This suggests that, in epithelial cells, some of the functions of p53 leading to cell-cycle arrest and apoptosis are restrained by HHV-6B infection, whereas other cellular defences, causing inhibition of virus transcription, are partially retained. PMID- 18420788 TI - Infection of neonates with murine gammaherpesvirus 68 results in enhanced viral persistence in lungs and absence of infectious mononucleosis syndrome. AB - We used the murine gammaherpesvirus 68 (gammaHV-68), which serves as a model for human gammaherpesvirus infection, to determine whether age at infection altered the pattern of gammaherpesvirus pathogenesis. We infected mice intranasally at 8 days old (pups) and 6 weeks old (adults) to investigate differences in gammaHV-68 pathogenesis. There was no difference between adults or pups in acute infection in the lungs at 6 days post-infection (p.i.). However, mice infected as pups exhibited a more disseminated viral infection with viral DNA detected in the spleen, liver and heart as measured by quantitative PCR (Q-PCR). In addition, viral DNA was detected in the lungs of mice infected as pups until 60 days p.i. Three viral transcripts (M2, M3 and M9) were expressed at both 30 and 60 days p.i. In contrast, no viral DNA or mRNA expression was detected in lungs of mice infected as adults at 30 or 60 days p.i. Mice infected as adults experienced a peak in latent infection in the spleen at 16 days p.i., corresponding with an increase in splenic weight and expansion of the Vbeta4+ CD8+ T-cell population, similar to infectious mononucleosis observed following infection of young adults with Epstein-Barr virus. However, the increase in splenic weight of infected pups was not as pronounced and no significant increase in Vbeta4+ CD8+ T-cell expansion was observed in infected pups. Together, these data suggest that the pathogenesis of murine gammaherpesvirus gammaHV-68 is age-dependent. PMID- 18420790 TI - Mouse cytomegalovirus inhibits beta interferon (IFN-beta) gene expression and controls activation pathways of the IFN-beta enhanceosome. AB - We have investigated beta interferon (IFN-beta) and IFN-alpha4 gene expression and activation of related transcription factors in mouse cytomegalovirus (MCMV) infected fibroblasts. mRNA analysis demonstrated an initial phase of IFN gene induction upon MCMV infection, which was followed by a sustained MCMV-mediated simultaneous downregulation of IFN-beta and IFN-alpha4 gene expression. The induction of IFN transcription resulted from the activation of the components of the IFN-beta enhanceosome, i.e. IFN regulatory factor (IRF) 3, nuclear factor (NF)-kappaB, activating transcription factor (ATF)-2 and c-Jun. Activation of the transcription factors occurred rapidly and in a sequential order upon infection, but only lasted a while. As a consequence, IFN-alpha/beta gene expression became undetectable 6 h post-infection and throughout the MCMV replication cycle. This effect is based on an active interference since restimulation of IFN gene induction by further external stimuli (e.g. Sendai virus infection) was completely abolished. This inhibition required MCMV gene expression and was not observed in cells infected with UV-inactivated MCMV virions. The efficiency of inhibition is achieved by a concerted blockade of IkappaBalpha degradation and a lack of nuclear accumulation of IRF3 and ATF-2/c-Jun. Using an MCMV mutant lacking pM27, a signal transducer and activator of transcription (STAT) 2 specific inhibitor of Jak/STAT signalling, we found that the initial phase of IFN induction and the subsequent inhibition does not depend on the positive-IFN feedback loop. Our findings indicate that the MCMV-mediated downregulation of IFN transcription in fibroblasts relies on a large arsenal of inhibitory mechanisms targeting each pathway that contributes to the multiprotein enhanceosome complex. PMID- 18420789 TI - Endoplasmic reticulum chaperones participate in human cytomegalovirus US2 mediated degradation of class I major histocompatibility complex molecules. AB - Inhibition of cell-surface expression of major histocompatibility complex class I molecules by human cytomegalovirus (HCMV, a beta-herpesvirus) promotes escape from recognition by CD8+ cytotoxic T cells. The HCMV US2 and US11 gene products induce class I downregulation during the early phase of HCMV infection by facilitating the degradation of class I heavy chains. The HCMV proteins promote the transport of the class I heavy chains across the endoplasmic reticulum (ER) membrane into the cytosol by a process referred to as 'dislocation', which is then followed by proteasome degradation. This process has striking similarities to the degradation of misfolded ER proteins mediated by ER quality control. Even though the major steps of the dislocation reaction have been characterized, the cellular proteins, specifically the ER chaperones involved in targeting class I for dislocation, have not been fully delineated. To elucidate the chaperones involved in HCMV-mediated class I dislocation, we utilized a chimeric class I heavy chain with an affinity tag at its carboxy terminus. Interestingly, US2 but not US11 continued to target the class I chimera for destruction, suggesting a structural limitation for US11-mediated degradation. Association studies in US2 cells and in cells that express a US2 mutant, US2-186HA, revealed that class I specifically interacts with calnexin, BiP and calreticulin. These findings demonstrate that US2-mediated class I destruction utilizes specific chaperones to facilitate class I dislocation. The data suggest a more general model in which the chaperones that mediate protein folding may also function during ER quality control to eliminate aberrant ER proteins. PMID- 18420791 TI - The conserved N-terminal domain of herpes simplex virus 1 UL24 protein is sufficient to induce the spatial redistribution of nucleolin. AB - UL24 is widely conserved among herpesviruses but its function during infection is poorly understood. Previously, we discovered a genetic link between UL24 and the herpes simplex virus 1-induced dispersal of the nucleolar protein nucleolin. Here, we report that in the absence of viral infection, transiently expressed UL24 accumulated in both the nucleus and the Golgi apparatus. In the majority of transfected cells, nuclear staining for UL24 was diffuse, but a minor staining pattern, whereby UL24 was present in nuclear foci corresponding to nucleoli, was also observed. Expression of UL24 correlated with the dispersal of nucleolin. This dispersal did not appear to be a consequence of a general disaggregation of nucleoli, as foci of fibrillarin staining persisted in cells expressing UL24. The conserved N-terminal region of UL24 was sufficient to cause this change in subcellular distribution of nucleolin. Interestingly, a bipartite nuclear localization signal predicted within the C terminus of UL24 was dispensable for nuclear localization. None of the five individual UL24 homology domains was required for nuclear or Golgi localization, but deletion of these domains resulted in the loss of nucleolin-dispersal activity. We determined that a nucleolar-targeting signal was contained within the first 60 aa of UL24. Our results show that the conserved N-terminal domain of UL24 is sufficient to specifically induce dispersal of nucleolin in the absence of other viral proteins or virus-induced cellular modifications. These results suggest that UL24 directly targets cellular factors that affect the composition of nucleoli. PMID- 18420792 TI - Antibodies against Epstein-Barr virus gp78 antigen: a novel marker for serological diagnosis of nasopharyngeal carcinoma detected by xMAP technology. AB - Immunoglobulin (Ig) A and/or IgG reactivities to several Epstein-Barr virus (EBV) antigens have been used to facilitate diagnosis of nasopharyngeal carcinoma (NPC). However, antibodies against gp78, an EBV membrane glycoprotein, have not been examined to this day. In this study, we utilized Luminex multi-analyte profiling (xMAP) technology to analyse antibody responses to a synthetic peptide of gp78 in sera samples from 95 NPC patients and 91 healthy controls. Our results showed the sensitivity and specificity of IgA-gp78 for NPC diagnosis were 79 and 71 %, respectively, while those of IgG-gp78 were 74 and 73 %, respectively. The IgA and IgG responses to different EBV antigens were not identical within an individual and IgA-gp78 and IgG-gp78 could be complementary to antibodies against viral capsid antigen (VCA), the diffused early antigen (EA-D) and the nuclear antigen EBNA1 for NPC diagnosis. When the six EBV parameters for NPC prediction, i.e. IgA-gp78, IgG-gp78, IgA-VCA, IgA-EBNA1, IgA-EA-D and IgG-EA-D, are combined, the combined predictors were able to reach overall sensitivity and specificity of 91 and 95 %, respectively. Thus, simultaneous detection of these EBV serological markers could improve the predictive values of NPC using xMAP technology. PMID- 18420793 TI - Molecular epidemiology of the African horse sickness virus S10 gene. AB - Between 2004 and 2006, 145 African horse sickness viruses (AHSV) were isolated from blood and organ samples submitted from South Africa to the Faculty of Veterinary Science, University of Pretoria. All nine serotypes were represented, with a range of 3-60 isolates per serotype. The RNA small segment 10 (S10) nucleotide sequences of these isolates were determined and the phylogeny investigated. AHSV, bluetongue virus (BTV) and equine encephalosis virus (EEV) all formed monophyletic groups and BTV was genetically closer to AHSV than EEV. This study confirmed the presence of three distinct S10 phylogenetic clades (alpha, beta and gamma). Some serotypes (6, 8 and 9 in alpha; 3 and 7 in beta; 2 in gamma) were restricted to a single clade, while other serotypes (1, 4 and 5) clustered into both the alpha and gamma clades. Strong purifying selection was evident and a constant molecular clock was inappropriate. The S10 gene is the second most variable gene of the AHSV genome and the use of S10 in molecular epidemiology was illustrated by an AHS outbreak in the Western Cape in 2004. It was shown that two separate AHSV were circulating in the area, even though AHSV serotype 1 was the only isolate from the outbreak. The small size of the gene (755-764 bp) and conserved terminal regions facilitate easy and quick sequencing. The establishment of an S10 sequence database is important for characterizing outbreaks of AHS. It will be an essential resource for elucidating the epidemiology of AHS. PMID- 18420794 TI - ORF018R, a highly abundant virion protein from Singapore grouper iridovirus, is involved in serine/threonine phosphorylation and virion assembly. AB - Iridovirus is an important pathogen causing serious diseases among wild, cultured and ornamental fish. Previous studies have shown that Singapore grouper iridovirus (SGIV) contains 162 open reading frames (ORFs) from which 51 viral proteins have been confirmed by proteomics studies. ORF018R, which is conserved among vertebrate iridoviruses, is an abundant virion protein identified from SGIV. Here, immunofluorescence staining showed that ORF018R occurred at high abundance throughout SGIV-infected cells. The function of ORF018R was explored using antisense morpholino oligonucleotides (asMOs). Knockdown of ORF018R expression resulted in a reduction in the expression of viral late genes, distortion of viral particle assembly and inhibition of SGIV infection in grouper embryonic cells. Western blotting with phosphoserine-specific antibody indicated that serine phosphorylation was significantly enhanced for proteins of molecular masss 17-32 kDa by SDS-PAGE when ORF018R expression was eliminated. These proteins were analysed further by two-dimensional gel electrophoresis, and numerous protein spots were found to shift to a lower pI and higher molecular mass as a result of the loss of ORF018R function. Five proteins with enhanced phosphorylation were identified by matrix-assisted laser desorption/ionization time-of-flight (TOF)-TOF mass spectrometry, including three viral proteins: ORF049L (dUTPase), ORF075R and ORF086R, and two host proteins: subunit 12 of eukaryotic translation factor 3 and natural killer enhancing factor. These findings suggest that ORF018R is involved in serine/threonine phosphorylation in SGIV-infected late-stage cells and plays an important role in expression of viral late genes and virion assembly. PMID- 18420795 TI - Supragenotypic groups of the hepatitis B virus genome. AB - Phylogenetic relationships among hepatitis B virus (HBV) genotypes were investigated using different regions across the genome. The phylogenetic analysis in conjunction with graphical examination of phylogenetic distance matrices and distance frequency distribution plotting suggest the clustering of HBV genotypes into three higher-order hierarchical groups: group I, comprising genotypes A-E and G; group II, comprising genotypes F and H; and a hypothetical group III. Present-day genotype G is postulated to be a recombinant with the non-polymerase region of group III virus and the polymerase gene of an ancestral virus belonging to group I. PMID- 18420796 TI - Hypermutation of hepatitis B virus genomes by APOBEC3G, APOBEC3C and APOBEC3H. AB - Hepatitis B virus (HBV) is a DNA virus that causes liver disease and replicates by reverse transcription of an RNA template. Previous studies have reported that HBV genomes bearing G-->A hypermutation are present at low frequency in human serum. These mutations are most likely due to the activity of apolipoprotein B mRNA-editing enzyme-catalytic polypeptide-like (APOBEC) cytosine deaminases, cellular proteins known to confer innate immunity against retroviruses by generating lethal hypermutations in viral genomes. This study assessed APOBEC3G, APOBEC3C and APOBEC3H, three members of this protein family present in human liver, for their ability to edit HBV genomes. Transfection of human HepG2 hepatoma cells with a plasmid encoding the APOBEC3C protein resulted in abundant G-->A mutations in the majority of newly formed HBV genomes. By contrast, transfection of APOBEC3G- and APOBEC3H-encoding plasmids only marginally increased hypermutation rates above the level caused by the cytosine deaminases naturally present in HepG2 cells. APOBEC3G- and APOBEC3H-mediated hypermutation, however, was clearly revealed by transfection of chicken LMH hepatoma cells, which lack endogenous cytosine deaminases. These results indicate that APOBEC3G, APOBEC3C and APOBEC3H have the ability to edit HBV DNA and that each protein is likely to contribute to various degrees to the generation of modified genomes in human liver cells. PMID- 18420797 TI - DNA vaccination in combination or not with lamivudine treatment breaks humoral immune tolerance and enhances cccDNA clearance in the duck model of chronic hepatitis B virus infection. AB - This study used a duck hepatitis B virus (DHBV) model to evaluate whether a novel DNA vaccination protocol alone or associated with antiviral (lamivudine) treatment was able to clear the intrahepatic covalently closed, circular viral DNA (cccDNA) pool responsible for persistence of infection. DHBV carriers received DNA vaccine (on weeks 6, 10, 13, 14, 28 and 35) targeting the large envelope and/or core proteins alone or combined with lamivudine treatment (on weeks 1-8) or lamivudine monotherapy. After 10 months of follow-up, a dramatic decrease in viraemia and liver DHBV cccDNA (below 0.08 cccDNA copies per cell) was observed in 9/30 ducks (30 %) receiving DNA mono- or combination therapy, compared with 0/12 (0 %) from lamivudine monotherapy or the control groups, suggesting a significant antiviral effect of DNA immunization. However, association with the drug did not significantly improve DHBV DNA vaccine efficacy (33 % cccDNA clearance for the combination vs 27 % for DNA monotherapy), probably due to the low antiviral potency of lamivudine in the duck model. Seroconversion to anti-preS was observed in 6/9 (67 %) ducks showing cccDNA clearance, compared with 1/28 (3.6 %) without clearance, suggesting a significant correlation (P<0.001) between humoral response restoration and cccDNA elimination. Importantly, an early (weeks 10-12) drop in viraemia was observed in seroconverted animals, and virus replication did not rebound following the cessation of immunotherapy, indicating a sustained effect. This study provides the first evidence that therapeutic DNA vaccination is able to enhance hepadnaviral cccDNA clearance, which is tightly associated with a break in humoral immune tolerance. These results also highlight the importance of antiviral drug potency and an effective DNA immunization protocol for the design of therapeutic vaccines against chronic hepatitis B. PMID- 18420798 TI - Analysis of the genome of Spodoptera frugiperda nucleopolyhedrovirus (SfMNPV-19) and of the high genomic heterogeneity in group II nucleopolyhedroviruses. AB - The genome of the most virulent among 22 Brazilian geographical isolates of Spodoptera frugiperda nucleopolyhedrovirus, isolate 19 (SfMNPV-19), was completely sequenced and shown to comprise 132,565 bp and 141 open reading frames (ORFs). A total of 11 ORFs with no homology to genes in the GenBank database were found. Of those, four had typical baculovirus promoter motifs and polyadenylation sites. Computer-simulated restriction enzyme cleavage patterns of SfMNPV-19 were compared with published physical maps of other SfMNPV isolates. Differences were observed in terms of the restriction profiles and genome size. Comparison of SfMNPV-19 with the sequence of the SfMNPV isolate 3AP2 indicated that they differed due to a 1427 bp deletion, as well as by a series of smaller deletions and point mutations. The majority of genes of SfMNPV-19 were conserved in the closely related Spodoptera exigua NPV (SeMNPV) and Agrotis segetum NPV (AgseMNPV A), but a few regions experienced major changes and rearrangements. Synthenic maps for the genomes of group II NPVs revealed that gene collinearity was observed only within certain clusters. Analysis of the dynamics of gene gain and loss along the phylogenetic tree of the NPVs showed that group II had only five defining genes and supported the hypothesis that these viruses form ten highly divergent ancient lineages. Crucially, more than 60 % of the gene gain events followed a power-law relation to genetic distance among baculoviruses, indicative of temporal organization in the gene accretion process. PMID- 18420799 TI - Bombyx mori nucleopolyhedrovirus ORF56 encodes an occlusion-derived virus protein and is not essential for budded virus production. AB - Bombyx mori nucleopolyhedrovirus ORF56 (Bm56) is a baculovirus core gene that is highly conserved in all baculoviruses that have had their genomes sequenced to date. Its transcripts in BmNPV-infected cells could be detected from 12 h post infection (p.i.) and the encoded protein could be detected at 16 h p.i. by using a polyclonal antibody against glutathione S-transferase-Bm56 fusion protein. Western blot analysis showed that Bm56 is a structural component of the occlusion derived virus nucleocapsid. Subsequent confocal microscopy revealed that Bm56 was distributed in the outer nuclear membrane and the intranuclear region of infected cells. To investigate the role of Bm56 in virus replication, a Bm56-knockout bacmid of BmNPV was constructed via homologous recombination in Escherichia coli. The Bm56 deletion had no effect on budded virus (BV) production in cultured cells; however, the deletion affected occlusion-body morphogenesis. A larval bioassay demonstrated that the Bm56 deletion did not reduce infectivity, whereas it resulted in a 50 % lethal time that was 16-18 h longer than that of the wild type bacmid at every dose used in this study. These results indicate that Bm56 facilitates efficient virus production in vivo; however, it is not essential for BV production in vitro. PMID- 18420800 TI - Low multiplicity of infection in vivo results in purifying selection against baculovirus deletion mutants. AB - The in vivo fate of Autographa californica multiple nucleopolyhedrovirus deletion mutants originating from serial passage in cell culture was investigated by passaging a population enriched in these mutants in insect larvae. The infectivity of polyhedra and occlusion-derived virion content per polyhedron were restored within two passages in vivo. The frequency of occurrence of deletion mutants was determined by real-time PCR. The frequency of the non-homologous region origin (non-HR ori) of DNA replication was reduced to wild-type levels within two passages. The frequency of the polyhedrin gene did not increase and remained below wild-type levels. A low m.o.i. during the initial infection in insect larvae, causing strong purifying selection for autonomously replicating viruses, could explain these observations. The same virus population used in vivo was also passaged once at a different m.o.i. in cell culture. A similar effect (i.e. lower non-HR ori frequency) was observed at low m.o.i. only, indicating that m.o.i. was the key selective condition. PMID- 18420801 TI - Activation of sterol regulatory element-binding protein 1c and fatty acid synthase transcription by hepatitis C virus non-structural protein 2. AB - Transcriptional factor sterol regulatory element-binding protein 1c (SREBP-1c) activates the transcription of lipogenic genes, including fatty acid synthase (FAS). Hepatitis C virus (HCV) infection is often associated with lipid accumulation within the liver, known as steatosis in the clinic. The molecular mechanisms of HCV-associated steatosis are not well characterized. Here, we showed that HCV non-structural protein 2 (NS2) activated SREBP-1c transcription in human hepatic Huh-7 cells as measured by using a human SREBP-1c promoter luciferase reporter plasmid. We further showed that sterol regulatory element (SRE) and liver X receptor element (LXRE) in the SREBP-1c promoter were involved in SREBP-1c activation by HCV NS2. Furthermore, expression of HCV NS2 resulted in the upregulation of FAS transcription. We also showed that FAS upregulation by HCV NS2 was SREBP-1-dependent since deleting the SRE sequence in a FAS promoter and expressing a dominant-negative SREBP-1 abrogated FAS promoter upregulation by HCV NS2. Taken together, our results suggest that HCV NS2 can upregulate the transcription of SREBP-1c and FAS, and thus is probably a contributing factor for HCV-associated steatosis. PMID- 18420802 TI - Hepatitis C virus NS5A protein interacts with and negatively regulates the non receptor protein tyrosine kinase Syk. AB - Hepatitis C virus (HCV) is the major causative agent of hepatocellular carcinoma. However, the precise mechanism underlying the carcinogenesis is yet to be elucidated. It has recently been reported that Syk, a non-receptor protein tyrosine kinase, functions as a potent tumour suppressor in human breast carcinoma. This study first examined the possible effect of HCV infection on expression of Syk in vivo. Immunohistochemical analysis revealed that endogenous Syk, which otherwise was expressed diffusely in the cytoplasm of normal hepatocytes, was localized near the cell membrane with a patchy pattern in HCV infected hepatocytes. The possible interaction between HCV proteins and Syk in human hepatoma-derived Huh-7 cells was then examined. Immunoprecipitation analysis revealed that NS5A interacted strongly with Syk. Deletion-mutation analysis revealed that an N-terminal portion of NS5A (aa 1-175) was involved in the physical interaction with Syk. An in vitro kinase assay demonstrated that NS5A inhibited the enzymic activity of Syk and that, in addition to the N terminal 175 residues, a central portion of NS5A (aa 237-302) was required for inhibition of Syk. Moreover, Syk-mediated phosphorylation of phospholipase C gamma1 was downregulated by NS5A. An interaction of NS5A with Syk was also detected in Huh-7.5 cells harbouring an HCV RNA replicon or infected with HCV. In conclusion, these results demonstrated that NS5A interacts with Syk resulting in negative regulation of its kinase activity. The results indicate that NS5A may be involved in the carcinogenesis of hepatocytes through the suppression of Syk kinase activities. PMID- 18420803 TI - Altered chemotactic response of myeloid and plasmacytoid dendritic cells from patients with chronic hepatitis C: role of alpha interferon. AB - Dendritic cell (DC) frequencies in the blood of patients with chronic hepatitis C virus (HCV) infection have been shown to be reduced significantly compared with those in healthy individuals. There is a further reduction of circulating myeloid DCs (MDCs) and plasmacytoid DCs (PDCs) in HCV patients receiving alpha interferon (IFN-alpha)-based antiviral therapy. Altered homing behaviour of DCs may be a possible mechanism for their 'loss' in peripheral blood in these clinical conditions. Systemic chemokine levels were measured by ELISA. Phenotypes and migratory properties of MDCs and PDCs from HCV patients were analysed by flow cytometry and chemotaxis assay. Compared with healthy controls, HCV patients had increased serum levels of inflammatory and constitutively expressed chemokines. Spontaneously generated MDCs from HCV patients were less mature, and both MDCs and PDCs showed intrinsic activation of receptors for inflammatory chemokines, thus suggesting an increased propensity to migrate towards inflammatory sites. IFN-alpha treatment in vitro induced MDC maturation and skewed the migratory response of both MDCs and PDCs towards chemokines expressed constitutively in secondary lymphoid organs. In conclusion, our results hint at altered homing behaviour of DCs during chronic HCV infection. IFN-alpha therapy may redirect DC migration from inflamed hepatic portal areas towards secondary lymphoid tissue. PMID- 18420804 TI - Multiple regions in dengue virus capsid protein contribute to nuclear localization during virus infection. AB - During infection, the capsid (C) protein of many flaviviruses localizes to the nuclei and nucleoli of several infected cell lines; the underlying basis and significance of C protein nuclear localization remain poorly understood. In this study, double alanine-substitution mutations were introduced into three previously proposed nuclear-localization signals (at positions 6-9, 73-76 and 85 100) of dengue virus C protein, and four viable mutants, c(K6A,K7A), c(K73A,K74A), c(R85A,K86A) and c(R97A,R98A), were generated in a mosquito cell line in which C protein nuclear localization was rarely observed. Indirect immunofluorescence analysis revealed that, whilst C protein was present in the nuclei of PS and Vero cells throughout infection with a dengue serotype 2 parent virus, the substitution mutations in c(K73A,K74A) and c(R85A,K86A) resulted in an elimination of nuclear localization in PS cells and marked reduction in Vero cells. Mutants c(K6A,K7A) and c(R97A,R98A) exhibited reduced nuclear localization at the late period of infection in PS cells only. All four mutants displayed reduced replication in PS, Vero and C6/36 cells, but there was a lack of correlation between nuclear localization and viral growth properties. Distinct dibasic residues within dengue virus C protein, many of which were located on the solvent-exposed side of the C protein homodimer, contribute to its ability to localize to nuclei during virus infection. PMID- 18420805 TI - Complete genome sequence analysis of Seneca Valley virus-001, a novel oncolytic picornavirus. AB - The complete genome sequence of Seneca Valley virus-001 (SVV-001), a small RNA virus, was determined and was shown to have typical picornavirus features. The 7280 nt long genome was predicted to contain a 5' untranslated region (UTR) of 666 nt, followed by a single long open reading frame consisting of 6543 nt, which encodes a 2181 aa polyprotein. This polyprotein could potentially be cleaved into 12 polypeptides in the standard picornavirus L-4-3-4 layout. A 3' UTR of 71 nt was followed by a poly(A) tail of unknown length. Comparisons with other picornaviruses showed that the P1, 2C, 3C and 3D polypeptides of SVV-001 were related most closely to those of the cardioviruses, although they were not related as closely to those of encephalomyocarditis virus and Theiler's murine encephalomyelitis virus as the latter were to each other. Most other regions of the polyprotein differed considerably from those of all other known picornaviruses. SVV-001 contains elements of an internal ribosome entry site reminiscent of that found in hepatitis C virus and a number of genetically diverse picornaviruses. SVV-001 is a novel picornavirus and it is proposed that it be classified as the prototype species in a novel genus named 'Senecavirus'. PMID- 18420806 TI - Complete genome analysis of hepatitis C virus subtypes 6t and 6u. AB - Hepatitis C virus (HCV) genomes exhibit high nucleotide sequence diversity. In this study, we performed complete genome sequence analysis of 11 HCV genotype 6 samples from Vietnam and Thailand. We identified nine HCV complete genomes belonging to subtypes 6a (D9), 6e (D42 and D88), 6f (TH52), 6i (TH24), 6l (D33), 6n (TH22 and TH31) and 6o (D85). Phylogenetic analysis of the core/E1 and NS5B regions from unclassified genotype 6 isolates from Asian immigrants in Canada revealed that two other viruses (D49 and D83) could be classified as novel candidates of HCV subtypes 6t and 6u. PMID- 18420807 TI - Genomic characterizations of bat coronaviruses (1A, 1B and HKU8) and evidence for co-infections in Miniopterus bats. AB - We previously reported the detection of bat coronaviruses (bat CoVs 1A, 1B, HKU7, HKU8 and bat-severe acute respiratory syndrome coronavirus) in Miniopterus spp. that cohabit a cave in Hong Kong. Here, we report the full genomic sequences of bat CoVs 1A, 1B and HKU8. Bat CoVs 1A and 1B, which are commonly found in the Miniopterus, are phylogenetically closely related. Using species-specific RT-PCR assays, bat CoVs 1A and 1B were confirmed to have distinct host specificities to Miniopterus magnater and Miniopterus pusillus, respectively. Interestingly, co infections of bat CoVs 1B and HKU8 in M. pusillus are detected in seven of 38 virus-positive specimens collected from 2004 to 2006. These findings highlight that co-infections of some coronaviruses might be common events in nature. The biological basis for the host restriction of bat coronaviruses, however, is yet to be determined. PMID- 18420808 TI - Modulation of the immune responses in chickens by low-pathogenicity avian influenza virus H9N2. AB - Most low-pathogenicity avian influenza (LPAI) viruses cause no or mild disease in avian species. Little is known about the mechanisms of host defence and the immune responses of avian influenza-infected birds. This study showed that chicken macrophages are susceptible to infection with LPAI H9N2 and H6N2 viruses and that infection led to apoptosis. In H9N2 virus-infected chicken macrophages, Toll-like receptor 7 responded to infection and mediated the cytokine responses. Whilst pro-inflammatory cytokines were largely upregulated, the interferon (IFN) response was fairly weak and IFN-inducible genes were differentially regulated. Among the regulated genes, major histocompatibility complex (MHC) antigens II were downregulated, which also occurred in the lungs of H9N2-infected chickens. Additionally, interleukin (IL)-4, IL-4 receptor and CD74 (MHC class II invariable chain) were also downregulated, all of which are pivotal in the activation of CD4+ helper T cells and humoral immunity. Remarkably, in H9N2 virus-infected chickens, the antibody response was severely suppressed. This was in contrast to the robust antibody response in chickens infected with H6N2 virus, in which expression of MHC class II antigens was upregulated. These data suggest that neutralizing antibodies and humoral immunity may not be developed efficiently in H9N2-infected chickens. These findings raise questions about how some LPAI viruses differentially regulate avian immune responses and whether they have similar effects on mammalian immune function. PMID- 18420809 TI - The C, V and W proteins of Nipah virus inhibit minigenome replication. AB - Nipah virus (NiV) is a recently emergent, highly pathogenic, zoonotic paramyxovirus of the genus Henipavirus. Like the phosphoprotein (P) gene of other paramyxoviruses, the P gene of NiV is predicted to encode three additional proteins, C, V and W. When the C, V and W proteins of NiV were tested for their ability to inhibit expression of the chloramphenicol acetyltransferase (CAT) reporter gene in plasmid-based, minigenome replication assays, each protein inhibited CAT expression in a dose-dependent manner. The C, V and W proteins of NiV also inhibited expression of CAT from a measles virus (MV) minigenome, but not from a human parainfluenzavirus 3 (hPIV3) minigenome. Interestingly, the C and V proteins of MV, which have previously been shown to inhibit MV minigenome replication, also inhibited NiV minigenome replication; however, they were not able to inhibit hPIV3 minigenome replication. In contrast, the C protein of hPIV3 inhibited minigenome replication of hPIV3, NiV and MV. Although there is very limited amino acid sequence similarity between the C, V and W proteins within the paramyxoviruses, the heterotypic inhibition of replication suggests that these proteins may share functional properties. PMID- 18420810 TI - Quasispecies dynamics and fixation of a synonymous mutation in hantavirus transmission. AB - RNA-dependent RNA polymerases, the key enzymes in replication of RNA viruses, have a low fidelity; thus, these viruses replicate as a swarm of mutants termed viral quasispecies. Constant generation of new mutations allows RNA viruses to adapt swiftly to a novel environment through selection of both pre-existing and de novo-generated genetic variants. Here, quasispecies dynamics were studied in vivo in controlled hantavirus transmission from experimentally infected to naive rodents through infested cage bedding. An elementary step of virus microevolution was apparent, as one synonymous mutation (A759G) repeatedly became fixed in the viral RNA quasispecies populations in the recipient animals. PMID- 18420811 TI - Identification of amino acids of the beet necrotic yellow vein virus p25 protein required for induction of the resistance response in leaves of Beta vulgaris plants. AB - The RNA3-encoded p25 protein of beet necrotic yellow vein virus (BNYVV) is responsible for the production of rhizomania symptoms of sugar beet roots (Beta vulgaris subsp. vulgaris). Here, it was found that the presence of the p25 protein is also associated with the resistance response in rub-inoculated leaves of sugar beet and wild beet (Beta vulgaris subsp. maritima) plants. The resistance phenotype displayed a range of symptoms from no visible lesions to necrotic or greyish lesions at the inoculation site, and only very low levels of virus and viral RNA accumulated. The susceptible phenotype showed large, bright yellow lesions and developed high levels of virus accumulation. In roots after Polymyxa betae vector inoculation, however, no drastic differences in virus and viral RNA accumulation levels were found between plants with susceptible and resistant phenotypes, except at an early stage of infection. There was a genotype specific interaction between BNYVV strains and two selected wild beet lines (MR1 and MR2) and sugar beet cultivars. Sequence analysis of natural BNYVV isolates and site-directed mutagenesis of the p25 protein revealed that 3 aa residues at positions 68, 70 and 179 are important in determining the resistance phenotype, and that host-genotype specificity is controlled by single amino acid changes at position 68. The mechanism of the occurrence of resistance-breaking BNYVV strains is discussed. PMID- 18420812 TI - PrP genotypes of free-ranging wapiti (Cervus elaphus nelsoni) with chronic wasting disease. AB - Variation in PrP prion gene sequence appears to modulate susceptibility to chronic wasting disease (CWD), a naturally occurring prion disease affecting four North American species of the family Cervidae. Wapiti (Cervus elaphus nelsoni) PrP is polymorphic at codon 132 [methionine (M) or leucine (L)]. We genotyped 171 samples, collected between 2002 and 2005 from CWD-infected and uninfected wapiti from three free-ranging populations in Colorado, USA, to study influences of PrP polymorphisms on CWD susceptibility further. Overall genotype frequencies for 124 apparently uninfected animals were 65.3 % MM132, 32.3 % ML132 and 2.4 % LL132; for 47 CWD-infected animals, these frequencies were 70.2 % MM132, 27.7 % ML132 and 2.1 % LL132. Surprisingly, our data revealed that, among recent (approx. 2002 -2005) CWD cases detected in free-ranging Colorado wapiti, the three PrP codon 132 genotypes were represented in proportion to their abundance in sampled populations (P> or =0.24) and all three genotypes showed equivalent susceptibility to infection. PMID- 18420813 TI - Effect of antimicrobial therapy on mortality in 377 episodes of Enterobacter spp. bacteraemia. AB - OBJECTIVES: The impact of appropriate antimicrobial therapy and antimicrobial resistance on the outcome of bacteraemia due to Enterobacter spp. remains unclear. The aim of our study was to evaluate the effect of antimicrobial therapy in 377 consecutive episodes of Enterobacter bacteraemia. PATIENTS AND METHODS: This includes retrospective analysis of a prospectively collected cohort. Clinical variables recorded were age, underlying diseases, use of corticosteroids, prognosis of underlying disease according to the McCabe and Jackson criteria, source of bacteraemia, need for mechanical ventilation, empirical antibiotic treatment, definitive treatment, antimicrobial susceptibility, presentation with septic shock and 30 day mortality rate. Univariate and multivariable analyses were performed to analyse the influence of antibiotic treatment and cephalosporin resistance on mortality. RESULTS: Between 1991 and 2006, 377 episodes of bacteraemia due to Enterobacter spp. (2.2%) were recorded. The frequency of Enterobacter bacteraemia significantly increased over these years. The overall mortality rate was 12.5% (47 of 377). Independent factors associated with 30 day mortality in patients with monomicrobial bacteraemia were rapidly fatal prognosis when compared with non-fatal prognosis, presentation with septic shock, patient under mechanical ventilation and unknown source of infection. The only factor independently associated with lower 30 day mortality was the empirical use of piperacillin/tazobactam. CONCLUSIONS: Enterobacter spp. are an increasing cause of bacteraemia. The empirical use of piperacillin/tazobactam was independently associated with a lower 30 day mortality rate. PMID- 18420814 TI - Misdiagnosis of serotonin syndrome as fibromyalgia and the role of physical therapists. AB - BACKGROUND AND PURPOSE: With increased use of serotonergic medications, a condition triggered by serotonin excess within the brain and spinal cord has emerged and may be gaining prevalence. The purposes of this case report are to describe how to identify serotonin syndrome in a patient who is taking citalopram (a selective serotonin reuptake inhibitor) on the basis of signs and symptoms and to promote the ability of physical therapists to recognize such signs and symptoms. CASE DESCRIPTION: The patient was a 42-year-old woman referred for physical therapy with a diagnosis of fibromyalgia. The physical therapist recognized that the patient's symptoms did not resemble those of fibromyalgia and recommended referral to a neurologist for further diagnostic testing. OUTCOMES: The patient was referred to a neurologist, who diagnosed serotonin syndrome related to the use of citalopram. The patient was weaned off citalopram and made a successful recovery, with scores on the Oswestry Disability Index decreasing from 70% to 28% at discharge from the physical therapy treatment and to 0% at the 6-month follow-up. The patient has since returned to her prior activity level, which includes skiing, motorcycle riding, and working at her consulting firm. DISCUSSION: This case report demonstrates how careful evaluation by the physical therapist indicated that signs and symptoms were not consistent with fibromyalgia, and further medical evaluation revealed the actual diagnosis of serotonin syndrome. PMID- 18420815 TI - TRIM21 is an IgG receptor that is structurally, thermodynamically, and kinetically conserved. AB - The newly identified tripartite motif (TRIM) family of proteins mediate innate immunity and other critical cellular functions. Here we show that TRIM21, which mediates the autoimmune diseases rheumatoid arthritis, systemic lupus erythematosus, and Sjogren's syndrome, is a previously undescribed IgG receptor with a binding mechanism unlike known mammalian Fcgamma receptors. TRIM21 simultaneously targets conserved hot-spot residues on both Ig domains of the Fc fragment using a PRYSPRY domain with a preformed multisite interface. The binding sites on both TRIM21 and Fc are highly conserved to the extent that the proteins are functionally interchangeable through murine, canine, primate, and human species. Pre-steady-state analysis exposes mechanistic conservation at the level of individual residues, which make the same energetic and kinetic contributions to binding despite varying in sequence. Together, our results reveal that TRIM21 is a previously undescribed type of IgG receptor based on a non-Ig scaffold whose interaction at the fundamental level-structural, thermodynamic, and kinetic-is evolutionarily conserved. PMID- 18420816 TI - Partitioned expression of duplicated genes during development and evolution of a single cell in a polyploid plant. AB - Polyploidy is an important driver of eukaryotic evolution, evident in many animals, fungi, and plants. One consequence of polyploidy is subfunctionalization, in which the ancestral expression profile becomes partitioned among duplicated genes (termed homoeologs). Subfunctionalization appears to be a common phenomenon insofar as it has been studied, at the scale of organs. Here, we use a high-resolution methodology to investigate the expression of thousands of pairs of homoeologs during the development of a single plant cell, using as a model the seed trichomes ("cotton fiber") of allopolyploid (containing "A" and "D" genomes) cotton (Gossypium). We demonstrate that approximately 30% of the homoeologs are significantly A- or D-biased at each of three time points studied during fiber development. Genes differentially biased toward the A or D genome belong to different biological processes, illustrating the functional partitioning of genomic contributions during cellular development. Interestingly, expression of the biased genes was shifted strongly toward the agronomically inferior D genome. Analyses of homoeologous gene expression during development of this cell showed that one-fifth of the genes exhibit changes in A/D ratios, indicating that significant alteration in duplicated gene expression is fairly frequent even at the level of development and maturation of a single cell. Comparing changes in homoeolog expression in cultivated versus wild cotton showed that most homoeolog expression bias reflects polyploidy rather than domestication. Evidence suggests, however, that domestication may increase expression bias in fibers toward the D genome, potentially implicating D-genome recruitment under human selection during domestication. PMID- 18420817 TI - Cardiogenic small molecules that enhance myocardial repair by stem cells. AB - The clinical success of stem cell therapy for myocardial repair hinges on a better understanding of cardiac fate mechanisms. We have identified small molecules involved in cardiac fate by screening a chemical library for activators of the signature gene Nkx2.5, using a luciferase knockin bacterial artificial chromosome (BAC) in mouse P19CL6 pluripotent stem cells. We describe a family of sulfonyl-hydrazone (Shz) small molecules that can trigger cardiac mRNA and protein expression in a variety of embryonic and adult stem/progenitor cells, including human mobilized peripheral blood mononuclear cells (M-PBMCs). Small molecule-enhanced M-PBMCs engrafted into the rat heart in proximity to an experimental injury improved cardiac function better than control cells. Recovery of cardiac function correlated with persistence of viable human cells, expressing human-specific cardiac mRNAs and proteins. Shz small molecules are promising starting points for drugs to promote myocardial repair/regeneration by activating cardiac differentiation in M-PBMCs. PMID- 18420818 TI - An assessment of fixed-capacity models of visual working memory. AB - Visual working memory is often modeled as having a fixed number of slots. We test this model by assessing the receiver operating characteristics (ROC) of participants in a visual-working-memory change-detection task. ROC plots yielded straight lines with a slope of 1.0, a tell-tale characteristic of all-or-none mnemonic representations. Formal model assessment yielded evidence highly consistent with a discrete fixed-capacity model of working memory for this task. PMID- 18420819 TI - From biosilicification to tailored materials: optimizing hydrophobic domains and resistance to protonation of polyamines. AB - Considerable research has been directed toward identifying the mechanisms involved in biosilicification to understand and possibly mimic the process for the production of superior silica-based materials while simultaneously minimizing pollution and energy costs. Molecules isolated from diatoms and, most recently sponges, thought to be key to this process contain polyamines with a propylamine backbone and variable levels of methylation. In a chemical approach to understanding the role of amine (especially propylamine) structures in silicification we have explored three key structural features: (i) the degree of polymerization, (ii) the level of amine methylation, and (iii) the size of the amine chain spacers. In this article, we show that there are two factors critical to their function: the ability of the amines to produce microemulsions and the presence of charged and uncharged amine groups within a molecule, with the latter feature helping to catalyze silicic acid condensation by a proton donor/acceptor mechanism. The understanding of amine-silicate interactions obtained from this study has enabled the controlled preparation of hollow and nonporous siliceous materials under mild conditions (circumneutral pH, room temperature, and in all aqueous systems) possibly compatible with the conditions used by biosystems. The "rules" identified from our study were further used predictively to modulate the activity of a given amine. We believe that the outcomes of the present contribution will form the basis for an approach to controlling the growth of inorganic materials by using tailor-made organic molecules. PMID- 18420820 TI - Sparse production but preferential incorporation of recently produced naive T cells in the human peripheral pool. AB - In mice, recent thymic emigrants (RTEs) make up a large part of the naive T cell pool and have been suggested to be a distinct short-lived pool. In humans, however, the life span and number of RTEs are unknown. Although (2)H(2)O labeling in young mice showed high thymic-dependent daily naive T cell production, long term up- and down-labeling with (2)H(2)O in human adults revealed a low daily production of naive T cells. Using mathematical modeling, we estimated human naive CD4 and CD8 T cell half-lives of 4.2 and 6.5 years, respectively, whereas memory CD4 and CD8 T cells had half-lives of 0.4 and 0.7 year. The estimated half life of recently produced naive T cells was much longer than these average half lives. Thus, our data are incompatible with a substantial short-lived RTE population in human adults and suggest that the few naive T cells that are newly produced are preferentially incorporated in the peripheral pool. PMID- 18420821 TI - Different phosphorylation states of the anaphase promoting complex in response to antimitotic drugs: a quantitative proteomic analysis. AB - The anaphase promoting complex (APC) controls the degradation of proteins during exit from mitosis and entry into S-phase. The activity of the APC is regulated by phosphorylation during mitosis. Because the phosphorylation pattern provides insights into the complexity of regulation of the APC, we studied in detail the phosphorylation patterns at a single mitotic state of arrest generated by various antimitotic drugs. We examined the phosphorylation patterns of the APC in HeLa S3 cells after they were arrested in prometaphase with taxol, nocodazole, vincristine, or monastrol. There were 71 phosphorylation sites on nine of the APC subunits. Despite the common state of arrest, the various antimitotic drug treatments resulted in differences in the phosphorylation patterns and phosphorylation stoichiometries. The relative phosphorylation stoichiometries were determined by using a method adapted from the isotope-free quantitation of the extent of modification (iQEM). We could show that during drug arrest the phosphorylation state of the APC changes, indicating that the mitotic arrest is not a static condition. We discuss these findings in terms of the variable efficacy of antimitotic drugs in cancer chemotherapy. PMID- 18420822 TI - Intrinsic noise, dissipation cost, and robustness of cellular networks: the underlying energy landscape of MAPK signal transduction. AB - We develop a probabilistic method for analyzing global features of a cellular network under intrinsic statistical fluctuations, which is important when there are finite numbers of molecules. By making a self-consistent mean field approximation of splitting the variables in order to reduce the large number of degrees of freedom, which is reasonable for a not very strongly interacting network, we discovered that the underlying energy landscape of the mitogen activated protein kinases (MAPKs) signal transduction network (with experimentally measured or inferred parameters such as chemical reaction rate coefficients in the network) is funneled toward a global minimum characterized by the nonequilibrium steady-state fixed point of the system at the end of the signal transduction process. For this system, we also show that the energy landscape is robust against intrinsic fluctuations and random perturbation to the inherent chemical reaction rates. The ratio of the slope versus the roughness of the energy landscape becomes a quantitative measure of robustness and stability of the network. Furthermore, we quantify the dissipation cost of this nonequilibrium system through entropy production, caused by the nonequilibrium flux in the system. We found that a lower dissipation cost corresponds to a more robust network. This least dissipation property might provide a design principle for robust and functional networks. Finally, we find the possibility of bistable and oscillatory-like solutions, which are important for cell fate decisions, upon perturbations. The method described here can be used in a variety of biological networks. PMID- 18420823 TI - The MUT9p kinase phosphorylates histone H3 threonine 3 and is necessary for heritable epigenetic silencing in Chlamydomonas. AB - Changes in chromatin organization are emerging as key regulators in nearly every aspect of DNA-templated metabolism in eukaryotes. Histones undergo many, largely reversible, posttranslational modifications that affect chromatin structure. Some modifications, such as trimethylation of histone H3 on Lys 4 (H3K4me3), correlate with transcriptional activation, whereas others, such as methylation of histone H3 on Lys 27 (H3K27me), are associated with silent chromatin. Posttranslational histone modifications may also be involved in the inheritance of chromatin states. Histone phosphorylation has been implicated in a variety of cellular processes but, because of the dynamic nature of this modification, its potential role in long-term gene silencing has remained relatively unexplored. We report here that a Chlamydomonas reinhardtii mutant defective in a Ser/Thr protein kinase (MUT9p), which phosphorylates histones H3 and H2A, shows deficiencies in the heritable repression of transgenes and transposons. Moreover, based on chromatin immunoprecipitation analyses, phosphorylated H3T3 (H3T3ph) and monomethylated H3K4 (H3K4me1) are inversely correlated with di/trimethylated H3K4 and associate preferentially with silenced transcription units. Conversely, the loss of those marks in mutant strains correlates with the transcriptional reactivation of transgenes and transposons. Our results suggest that H3T3ph and H3K4me1 function as reinforcing epigenetic marks for the silencing of euchromatic loci in Chlamydomonas. PMID- 18420824 TI - Selective disruption of the mammalian secretory apparatus enhances or eliminates calcium current modulation in nerve endings. AB - Modulation of secretion via G protein-coupled receptors (GPCRs) serves an important regulatory function in neuronal and nonneuronal secretory cells. Most secretory cells possess voltage-gated calcium channels, share homologues of the core complex of three proteins (the SNAREs) that constitute the secretory apparatus, and are modulated by GPCR activation. Activators of GPCRs generally inhibit the release of neurotransmitter substances to a maximum of only 50-60% of the control level, suggesting that complex protein-protein interactions may govern the efficacy of this form of modulation. In this article, molecular genetic approaches are used in combination with botulinum toxins (selective molecular scalpels that cleave the SNAREs at highly restricted loci) to address this issue. The results suggest that the cleavage of either of the plasma membrane SNAREs (syntaxin or SNAP-25) prevents modulation of calcium currents by A(1) adenosine receptors at mammalian motor nerve endings. In contrast, cleavage of the synaptic vesicle SNARE (synaptobrevin) in conjunction with deletion of the vesicle-docking protein Rab3A greatly enhances the efficacy of calcium current modulation. PMID- 18420825 TI - Subdiaphragmatic vagal afferent nerves modulate visceral pain. AB - Activation of the vagal afferents by noxious gastrointestinal stimuli suggests that vagal afferents may play a complex role in visceral pain processes. The contribution of the vagus nerve to visceral pain remains unresolved. Previous studies reported that patients following chronic vagotomy have lower pain thresholds. The patient with irritable bowel syndrome has been shown alteration of vagal function. We hypothesize that vagal afferent nerves modulate visceral pain. Visceromotor responses (VMR) to graded colorectal distension (CRD) were recorded from the abdominal muscles in conscious rats. Chronic subdiaphragmatic vagus nerve sections induced 470, 106, 51, and 54% increases in VMR to CRD at 20, 40, 60 and 80 mmHg, respectively. Similarly, at light level of anesthesia, topical application of lidocaine to the subdiaphragmatic vagus nerve in rats increased VMR to CRD. Vagal afferent neuronal responses to low or high-intensity electrical vagal stimulation (EVS) of vagal afferent Adelta or C fibers were distinguished by calculating their conduction velocity. Low-intensity EVS of Adelta fibers (40 microA, 20 Hz, 0.5 ms for 30 s) reduced VMR to CRD at 40, 60, and 80 mmHg by 41, 52, and 58%, respectively. In contrast, high-intensity EVS of C fibers (400 microA, 1 Hz, 0.5 ms for 30 s) had no effect on VMR to CRD. In conclusion, we demonstrated that vagal afferent nerves modulate visceral pain. Low-intensity EVS that activates vagal afferent Adelta fibers reduced visceral pain. Thus EVS may potentially have a role in the treatment of chronic visceral pain. PMID- 18420826 TI - CFTR inhibition augments NHE3 activity during luminal high CO2 exposure in rat duodenal mucosa. AB - We hypothesized that the function of duodenocyte apical membrane acid-base transporters are essential for H(+) absorption from the lumen. We thus examined the effect of inhibition of Na(+)/H(+) exchanger-3 (NHE3), cystic fibrosis transmembrane regulator (CFTR), or apical anion exchangers on transmucosal CO(2) diffusion and HCO(3)(-) secretion in rat duodenum. Duodena were perfused with a pH 6.4 high CO(2) solution or pH 2.2 low CO(2) solution with the NHE3 inhibitor, S3226, the anion transport inhibitor, DIDS, or pretreatment with the potent CFTR inhibitor, CFTR(inh)-172, with simultaneous measurements of luminal and portal venous (PV) pH and carbon dioxide concentration ([CO(2)]). Luminal high CO(2) solution increased CO(2) absorption and HCO(3)(-) secretion, accompanied by PV acidification and PV Pco(2) increase. During CO(2) challenge, CFTR(inh)-172 induced HCO(3)(-) absorption, while inhibiting PV acidification. S3226 reversed CFTR(inh)-associated HCO(3)(-) absorption. Luminal pH 2.2 challenge increased H(+) and CO(2) absorption and acidified the PV, inhibited by CFTR(inh)-172 and DIDS, but not by S3226. CFTR inhibition and DIDS reversed HCO(3)(-) secretion to absorption and inhibited PV acidification during CO(2) challenge, suggesting that HCO(3)(-) secretion helps facilitate CO(2)/H(+) absorption. Furthermore, CFTR inhibition prevented CO(2)-induced cellular acidification reversed by S3226. Reversal of increased HCO(3)(-) loss by NHE3 inhibition and reduced intracellular acidification during CFTR inhibition is consistent with activation or unmasking of NHE3 activity by CFTR inhibition, increasing cell surface H(+) available to neutralize luminal HCO(3)(-) with consequent CO(2) absorption. NHE3, by secreting H(+) into the luminal microclimate, facilitates net transmucosal HCO(3)(-) absorption with a mechanism similar to proximal tubular HCO(3)(-) absorption. PMID- 18420827 TI - Defective circulating CD25 regulatory T cells in patients with chronic immune thrombocytopenic purpura. AB - Immune thrombocytopenic purpura (ITP) is characterized by the presence of antiplatelet autoantibodies as a result of loss of tolerance. CD4+CD25+ regulatory T cells (Tregs) are important for maintenance of peripheral tolerance. Decreased levels of peripheral Tregs in patients with ITP have been reported. To test whether inefficient production or reduced immunosuppressive activity of Tregs contributes to loss of tolerance in patients with chronic ITP, we investigated the frequency and function of their circulating CD4+CD25(hi) Tregs. We found a com-parable frequency of circulating CD4+CD25(hi)Foxp3+ Tregs in patients and controls (n = 16, P > .05). However, sorted CD4+CD25(hi) cells from patients with chronic ITP (n = 13) had a 2-fold reduction of in vitro immunosuppressive activity compared with controls (n = 10, P < .05). The impaired suppression was specific to Tregs as shown by cross-mixing experiments with T cells from controls. These data suggest that functional defects in Tregs contribute to breakdown of self-tolerance in patients with chronic ITP. PMID- 18420828 TI - Neutrophil stress and apoptosis underlie myeloid dysfunction in glycogen storage disease type Ib. AB - Glycogen storage disease type Ib (GSD-Ib) is caused by a deficiency in the glucose-6-phosphate (G6P) transporter (G6PT) that works with a liver/kidney/intestine-restricted glucose-6-phosphatase-alpha (G6Pase-alpha) to maintain glucose homeostasis between meals. Clinically, GSD-Ib patients manifest disturbed glucose homeostasis and neutrophil dysfunctions but the cause of the latter is unclear. Neutrophils express the ubiquitously expressed G6PT and G6Pase beta that together transport G6P into the endoplasmic reticulum (ER) lumen and hydrolyze it to glucose. Because we expected G6PT-deficient neutrophils to be unable to produce endogenous glucose, we hypothesized this would lead to ER stress and increased apoptosis. Using GSD-Ib mice, we showed that GSD-Ib neutrophils exhibited increased production of ER chaperones and oxidative stress, consistent with ER stress, increased annexin V binding and caspase-3 activation, consistent with an increased rate of apoptosis. Bax activation, mitochondrial release of proapoptotic effectors, and caspase-9 activation demonstrated the involvement of the intrinsic mitochondrial pathway in these processes. The results demonstrate that G6P translocation and hydrolysis are required for normal neutrophil functions and support the hypothesis that neutrophil dysfunction in GSD-Ib is due, at least in part, to ER stress and increased apoptosis. PMID- 18420829 TI - Preliminary reliability and validity of the grandparent version of the Grandparent Support Scale for Teenage Mothers (GSSTM-G). AB - This study demonstrates the preliminary reliability and validity of the grandparent version of the Grandparent Support Scale for Teenage Mothers (GSSTM G), a measure designed to assess grandparents' perceptions of triadic relationships that support or imperil the teen mother's care of her baby. Grandparents (N = 61) who participated in this study were parents and parent surrogates of teen mothers who completed a teen version of the scale (GSSTM-T). Principal axis factoring with oblique rotation resulted in a reduction of the GSSTM-G scale from 19 to 14 items. A two-factor solution was supported by item factor loadings and conceptual clarity. The GSSTM-G factors revealed two different patterns of family relationships, Responsive Family Relationships (alpha = .82) and Adversarial Family Relationships (alpha = .81), that correlated with other measures. Construct validity and internal consistency support further exploration of the GSSTM-G as a measure of family relationships, as related to the care of the infant, from grandparents' perspectives. PMID- 18420830 TI - Lineage selection of functional and cryopreservable human embryonic stem cell derived neurons. AB - A major prerequisite for the biomedical application of human embryonic stem cells (hESC) is the derivation of defined and homogeneous somatic cell types. Here we present a human doublecortin (DCX) promoter-based lineage-selection strategy for the generation of purified hESC-derived immature neurons. After transfection of hESC-derived neural precursors with a DCX-enhanced green fluorescent protein construct, fluorescence-activated cell sorting enables the enrichment of immature human neurons at purities of up to 95%. Selected neurons undergo functional maturation and are able to establish synaptic connections. Considering that the applicability of purified hESC-derived neurons would largely benefit from an efficient cryopreservation technique, we set out to devise defined freezing conditions involving caspase inhibition, which yield post-thaw recovery rates of up to 83%. Combined with our lineage-selection procedure this cryopreservation technique enables the generation of human neurons in a ready-to-use format for a large variety of biomedical applications. PMID- 18420831 TI - Novel cardiac precursor-like cells from human menstrual blood-derived mesenchymal cells. AB - Stem cell therapy can help repair damaged heart tissue. Yet many of the suitable cells currently identified for human use are difficult to obtain and involve invasive procedures. In our search for novel stem cells with a higher cardiomyogenic potential than those available from bone marrow, we discovered that potent cardiac precursor-like cells can be harvested from human menstrual blood. This represents a new, noninvasive, and potent source of cardiac stem cell therapeutic material. We demonstrate that menstrual blood-derived mesenchymal cells (MMCs) began beating spontaneously after induction, exhibiting cardiomyocyte-specific action potentials. Cardiac troponin-I-positive cardiomyocytes accounted for 27%-32% of the MMCs in vitro. The MMCs proliferated, on average, 28 generations without affecting cardiomyogenic transdifferentiation ability, and expressed mRNA of GATA-4 before cardiomyogenic induction. Hypothesizing that the majority of cardiomyogenic cells in MMCs originated from detached uterine endometrial glands, we established monoclonal endometrial gland derived mesenchymal cells (EMCs), 76%-97% of which transdifferentiated into cardiac cells in vitro. Both EMCs and MMCs were positive for CD29, CD105 and negative for CD34, CD45. EMCs engrafted onto a recipient's heart using a novel 3 dimensional EMC cell sheet manipulation transdifferentiated into cardiac tissue layer in vivo. Transplanted MMCs also significantly restored impaired cardiac function, decreasing the myocardial infarction (MI) area in the nude rat model, with tissue of MMC-derived cardiomyocytes observed in the MI area in vivo. Thus, MMCs appear to be a potential novel, easily accessible source of material for cardiac stem cell-based therapy. PMID- 18420832 TI - Transforming growth factor beta cooperates with persephin for dopaminergic phenotype induction. AB - The aim of the present study was to investigate the putative cooperative effects of transforming growth factor beta (TGF-beta) and glial cell line-derived neurotrophic factor (GDNF) family ligands in the differentiation of midbrain progenitors toward a dopaminergic phenotype. Therefore, a mouse midbrain embryonic day (E) 12 neurospheres culture was used as an experimental model. We show that neurturin and persephin (PSPN), but not GDNF, are capable of transient induction of dopaminergic neurons in vitro. This process, however, requires the presence of endogenous TGF-beta. In contrast, after 8 days in vitro GDNF rescued the TGF-beta neutralization-dependent loss of the TH-positive cells. In vivo, at E14.5, no apparent phenotype concerning dopaminergic neurons was observed in Tgf beta2(-/-)/gdnf(-/-) double mutant mice. In vitro, combined TGF-beta/PSPN treatment achieved a yield of approximately 20% TH-positive cells that were less vulnerable against 1-methyl-4-phenyl pyridinium ion toxicity. The underlying TGF beta/PSPN differentiation signaling is receptor-mediated, involving p38 mitogen activated protein kinase and phosphatidylinositol 3-kinase pathways. These results indicate that phenotype induction and survival of fully differentiated neurons are accomplished through distinct pathways and individual factor requirement. TGF-beta is required for the induction of dopaminergic neurons, whereas GDNF is required for regulating and/or maintaining a differentiated neuronal phenotype. Moreover, this study suggests that the combination of TGF beta with PSPN is a potent inductive cocktail for the generation of dopaminergic neurons that should be considered in tissue engineering and cell replacement therapies for Parkinson's disease. PMID- 18420833 TI - Bone marrow-derived mesenchymal stem cells ameliorate autoimmune enteropathy independently of regulatory T cells. AB - Cell-based tolerogenic therapy is a relatively new approach for the treatment of autoimmune diseases. Mesenchymal stem cells (MSCs) have been shown to be potent immunomodulatory agents in a number of experimental and clinical scenarios; however, their use in various autoimmune diseases is undefined. Herein, we report the efficacy of MSC transplantation in a multiorgan autoimmunity model. Mice with defective peripheral tolerance caused by a deficiency in regulatory T cells were used as a testbed for therapy. After screening multiple target tissues of autoimmune attack, we observed an MSC-specific improvement in the histopathology of the distal ileum of treated mice. We then showed that MSCs can reduce mesenteric lymph node (MLN) cellularity in autoimmune mice during active disease and decrease activated T-cell populations in the MLN. Trafficking studies using enhanced green fluorescent protein (eGFP)-reporter MSCs revealed no appreciable engraftment in the intestine, but it did reveal the presence of eGFP+ cells organized in clusters within the MLN, as well as ancillary nodes. Semiquantitative analysis showed no difference in the number of clusters; however, eGFP+ cells in MLNs compared with ancillary nodes had distinct fibroblastoid morphology and formed a network with neighboring eGFP+ cells. Finally, we show evidence that transplantation of MSCs caused global immunosuppression, as measured by increased CD4+ CD8+ thymocyte production and serum interleukin-10 and decreased serum interferon-gamma. These data implicate the intestine as a new site of MSC tolerance induction and should motivate additional studies evaluating the use of MSCs as a treatment for autoimmune enteropathies. PMID- 18420834 TI - Transplantation of bone marrow-derived very small embryonic-like stem cells attenuates left ventricular dysfunction and remodeling after myocardial infarction. AB - Adult bone marrow (BM) contains Sca-1+/Lin-/CD45- very small embryonic-like stem cells (VSELs) that express markers of several lineages, including cardiac markers, and differentiate into cardiomyocytes in vitro. We examined whether BM derived VSELs promote myocardial repair after a reperfused myocardial infarction (MI). Mice underwent a 30-minute coronary occlusion followed by reperfusion and received intramyocardial injection of vehicle (n= 11), 1 x 10(5) Sca-1+/Lin /CD45+ enhanced green fluorescent protein (EGFP)-labeled hematopoietic stem cells (n= 13 [cell control group]), or 1 x 10(4) Sca-1+/Lin-/CD45- EGFP-labeled cells (n= 14 [VSEL-treated group]) at 48 hours after MI. At 35 days after MI, VSEL treated mice exhibited improved global and regional left ventricular (LV) systolic function (echocardiography) and attenuated myocyte hypertrophy in surviving tissue (histology and echocardiography) compared with vehicle-treated controls. In contrast, transplantation of Sca-1+/Lin-/CD45+ cells failed to confer any functional or structural benefits. Scattered EGFP+ myocytes and capillaries were present in the infarct region in VSEL-treated mice, but their numbers were very small. These results indicate that transplantation of a relatively small number of CD45- VSELs is sufficient to improve LV function and alleviate myocyte hypertrophy after MI, supporting the potential therapeutic utility of these cells for cardiac repair. Disclosure of potential conflicts of interest is found at the end of this article. PMID- 18420835 TI - Feasibility and reliability of the annual profile method for deriving QALYs for short-term health conditions. AB - INTRODUCTION: When health varies over time, the standard quality-adjusted life year model operates under the assumptions of time utility independence within each health state and additive independence between health states. These assumptions can be relaxed by an integral assessment of disease severity over time. The authors present the annual profile method (APM), which values health profiles on a 1-year base, and test the APM for feasibility, consistency, and test-retest reliability. METHODS: A population panel, general practitioners, medical advisers, and a panel of the Dutch Consumers Association valued vignettes for 46 disease stages using the visual analog scale (VAS) and time tradeoff (TTO) methods. Vignettes contained disease-specific information, a generic description (EQ-6D5L), a description of the disease course over time, and a visual representation of the disease. Feasibility was tested by missing and inconsistent responses. Consistency between and within panels was tested with a generalizability study, analysis of variance, and standard correlation coefficients. Test-retest reliability was tested with a generalizability study and intra-class correlation coefficients. RESULTS: Missing and inconsistent responses were < 2.6%. The valuations were consistent across panels, with generalizability coefficients of 0.78 (VAS) and 0.64 (TTO). Within the main population panel, internal consistency was satisfactory and the influence of background characteristics negligible. Test-retest reliability was high, with generalizability coefficients of 0.90 (VAS) and 0.72 (TTO). CONCLUSION: Feasibility and reliability of the APM for realistic health profiles are good to excellent. The APM is a promising step to bridge the gap between the quality adjusted life year methodology and clinical reality. PMID- 18420836 TI - Charting plasticity in the regenerating maps of the mammalian olfactory bulb. AB - The anatomical organization of a neural system can offer a glimpse into its functional logic. The basic premise is that by understanding how something is put together one can figure out how it works. Unfortunately, organization is not always represented purely at an anatomical level and is sometimes best revealed through molecular or functional studies. The mammalian olfactory system exhibits organizational features at all these levels including 1) anatomically distinct structural layers in the olfactory bulb, 2) molecular maps based upon odorant receptor expression, and 3) functional local circuits giving rise to odor columns that provide a contextual logic for an intrabulbar map. In addition, various forms of cellular plasticity have been shown to play an integral role in shaping the structural properties of most neural systems and must be considered when assessing each system's anatomical organization. Interestingly, the olfactory system invokes an added level of complexity for understanding organization in that it regenerates both at the peripheral and the central levels. Thus, olfaction offers a rare opportunity to study both the structural and the functional properties of a regenerating sensory system in direct response to environmental stimuli. In this review, we discuss neural organization in the form of maps and explore the relationship between regeneration and plasticity. PMID- 18420837 TI - Changes in Clara cell 10 kDa protein (CC10)-positive cell distribution in acute lung injury following repeated lipopolysaccharide challenge in the rat. AB - Clara cell 10 kDa protein (CC10) is the major secretory protein of Clara cells and is thought to play a protective role in the lung owing to its anti inflammatory properties. There is little information on the anatomical distribution of CC10-positive cells in rat lung following lipopolysaccharide (LPS) challenge. We have determined the expression of CC10 along the tracheobronchial tree in saline-treated and LPS-treated rats. Saline-treated rats showed sporadic CC10 staining in central airways and abundant staining in bronchioles. In transitional airways, most cells were positive except for squamous cells. Following LPS challenge, there was a reduction in staining in the upper airways but little change within bronchioles. Squamous epithelia within the transitional airways now showed positive staining. These cells also co-stained for pancytokeratin and appeared to co-localize with surfactant D- and Ki67 positive cells, indicating the presence of a dedifferentiated cell type with both epithelial and pneumocyte phenotypes. These data show that diffuse inflammatory injury results in generalized loss of CC10 in central airways. Conversely, the transitional airways showed evidence of a dedifferentiated population of squamous cells that now stained for CC10. We hypothesize that this is an attempt by peripheral lung to maintain alveolar sac integrity during an inflammatory episode. PMID- 18420838 TI - Family caregivers' strains: comparative analysis of cancer caregiving with dementia, diabetes, and frail elderly caregiving. AB - OBJECTIVE: This study aimed to investigate the impact of cancer from family caregivers' perspective, based on a comparative analysis of caregiving burden and distress, among four types of caregivers. METHODS: The sample included caregivers of persons with cancer, dementia, diabetes, or frail elderly from a nationally representative sample of 606 caregivers. RESULTS: Although the four different types of caregivers were comparable in most sociodemographic characteristics, caregiving involvement and caregiving outcomes differed among the caregiving groups. Both cancer and dementia caregivers reported greater levels of physical burden and psychological distress than other caregivers, after controlling for sociodemographic and caregiving involvement (i.e., level of burden and caregiving duration) factors. DISCUSSION: The comparative analysis provided a systematic review of cancer caregiving in the context of three other types of caregiving. Given the high levels of burden and distress, greater emphasis should be placed on developing social service policy and practice for cancer caregivers. PMID- 18420839 TI - Lipid rescue: a step forward in patient safety? Likely so! PMID- 18420840 TI - Lipid emulsion for the treatment of local anesthetic toxicity: patient safety implications. PMID- 18420841 TI - Lipid infusion therapy: translation to clinical practice. PMID- 18420842 TI - Haloperidol for postoperative nausea and vomiting: are we reinventing the wheel? PMID- 18420843 TI - Emulsified intravenous versus evaporated inhaled isoflurane for heart protection: old wine in a new bottle or true innovation? PMID- 18420844 TI - Emulsified isoflurane produces cardiac protection after ischemia-reperfusion injury in rabbits. AB - BACKGROUND: In this study, we examined the cardioprotective effects of parental emulsified isoflurane compared with inhaled isoflurane. METHODS: Thirty-two rabbits were subjected to 30 min of myocardial ischemia induced by temporary ligation of the left anterior descending coronary artery followed by 3 h of reperfusion. Before left anterior descending coronary artery occlusion, the rabbits were randomly allocated into one of four groups (eight for each group): group C, no ischemia preconditioning treatment; group IS, inhaled isoflurane 1.1% end-tidal; group EI, a continuous infusion of 8% emulsified isoflurane to an end tidal concentration of 0.64%; and group IN, a continuous infusion of 30% Intralipid started 30 min. Treatments were started 30 min before ischemia followed by a 15 min washout period for isoflurane groups. Myocardial infarct volume, lactate dehydrogenase, and creatine kinase levels were measured and changes in mitochondrial ultrastructure assessed after 3 h myocardial reperfusion. RESULTS: Myocardial infarct size 3 h after reperfusion was lower in groups IS and EI compared with groups C and IN (20% +/- 8%, 18% +/- 8%, 39% +/- 6%, and 34% +/- 9%, respectively, P < 0.01). There were no differences in myocardial infarct size between groups IS and EI or between groups C and IN. Plasma lactate dehydrogenase and creatine kinase levels were lower in group IS (456 +/- 58 U/L and 1725 +/- 230 U/L) and group EI (451 +/- 54 U/L and 1686 +/- 444 U/L) 3 h after myocardial reperfusion compared with groups C (676 +/- 82 U/L and 2373 +/- 529 U/L; P < 0.01). Mitochondrial ultrastructure changes were less pronounced in groups IS and EI compared with group C. CONCLUSIONS: Our results indicate that, in rabbits, i.v. emulsified isoflurane provides similar myocardial protection against ischemia-reperfusion injury as inhaled isoflurane. PMID- 18420845 TI - The in vitro effects of fibrinogen concentrate, factor XIII and fresh frozen plasma on impaired clot formation after 60% dilution. AB - BACKGROUND: Previous investigations have shown that increasing fibrinogen concentration improves dilution-dependent impairment of clot formation. We conducted an in vitro study to explore whether substitution with fibrin stabilizing factor XIII (FXIII) combined with fibrinogen promotes further improvement of clot formation, and whether fibrinogen administration as concentrate or fresh frozen plasma (FFP) results in comparable effects. METHODS: Blood from six healthy donors was diluted by 60% using lactated Ringer's solution. Aliquots of diluted blood samples were incubated with two different doses of fibrinogen concentrate, FXIII concentrate, the combination of both, or with two different doses of FFP. Using thrombelastometry (ROTEM) blood samples were analyzed at baseline (undiluted), after dilution and after supplementation. Variables were analyzed for changes from baseline, and effects of fibrinogen concentrate alone or combined with FXIII were compared with effects observed with corresponding FFP doses. RESULTS: After 60% in vitro dilution of blood all ROTEM parameters and global coagulation tests changed significantly. Among the substitutes tested FXIII alone had no effect, the combination with fibrinogen improved coagulation time, alpha angle and fibrinogen/fibrin polymerization significantly more than did small-dose fibrinogen alone. After substituting fibrinogen, median values of all ROTEM variables were within the normal range, thereby showing dose dependency but also significant differences (P = 0.027) from corresponding FFP doses (EXTEM MCF FFP small dose [38 (35, 40.3) mm)], which enabled only coagulation time to be shortened to baseline levels. CONCLUSIONS: Supplementation of fibrinogen restored all ROTEM parameters after dilution. This effect was partially enhanced by adding FXIII and was significantly stronger than for FFP substitution. PMID- 18420846 TI - Coagulation monitoring: current techniques and clinical use of viscoelastic point of-care coagulation devices. AB - Perioperative monitoring of blood coagulation is critical to better understand causes of hemorrhage, to guide hemostatic therapies, and to predict the risk of bleeding during the consecutive anesthetic or surgical procedures. Point-of-care (POC) coagulation monitoring devices assessing the viscoelastic properties of whole blood, i.e., thrombelastography, rotation thrombelastometry, and Sonoclot analysis, may overcome several limitations of routine coagulation tests in the perioperative setting. The advantage of these techniques is that they have the potential to measure the clotting process, starting with fibrin formation and continue through to clot retraction and fibrinolysis at the bedside, with minimal delays. Furthermore, the coagulation status of patients is assessed in whole blood, allowing the plasmatic coagulation system to interact with platelets and red cells, and thereby providing useful additional information on platelet function. Viscoelastic POC coagulation devices are increasingly being used in clinical practice, especially in the management of patients undergoing cardiac and liver surgery. Furthermore, they provide useful information in a large variety of clinical scenarios, e.g., massive hemorrhage, assessment of hypo- and hypercoagulable states, guiding pro- and anticoagulant therapies, and in diagnosing of a surgical bleeding. A surgical etiology of bleeding has to be considered when viscoelastic test results are normal. In summary, viscoelastic POC coagulation devices may help identify the cause of bleeding and guide pro- and anticoagulant therapies. To ensure optimal accuracy and performance, standardized procedures for blood sampling and handling, strict quality controls and trained personnel are required. PMID- 18420847 TI - Guidelines for the performance of a comprehensive intraoperative epiaortic ultrasonographic examination: recommendations of the American Society of Echocardiography and the Society of Cardiovascular Anesthesiologists; endorsed by the Society of Thoracic Surgeons. PMID- 18420848 TI - Ventriculoseptal defect detected after surgical repair of double-chamber right ventricle. PMID- 18420849 TI - Imaging a large unsuspected sinus of Valsalva aneurysm dissecting into the interventricular septum. PMID- 18420850 TI - Do new anesthesia ventilators deliver small tidal volumes accurately during volume-controlled ventilation? AB - BACKGROUND: During mechanical ventilation of infants and neonates, small changes in tidal volume may lead to hypo- or hyperventilation, barotrauma, or volutrauma. Partly because breathing circuit compliance and fresh gas flow affect tidal volume delivery by traditional anesthesia ventilators in volume-controlled ventilation (VCV) mode, pressure-controlled ventilation (PCV) using a circle breathing system has become a common approach to minimizing the risk of mechanical ventilation for small patients, although delivered tidal volume is not assured during PCV. A new generation of anesthesia machine ventilators addresses the problems of VCV by adjusting for fresh gas flow and for the compliance of the breathing circuit. In this study, we evaluated the accuracy of new anesthesia ventilators to deliver small tidal volumes. METHODS: Four anesthesia ventilator systems were evaluated to determine the accuracy of volume delivery to the airway during VCV at tidal volume settings of 100, 200, and 500 mL under different conditions of breathing circuit compliance (fully extended and fully contracted circuits) and lung compliance. A mechanical test lung (adult and infant) was used to simulate lung compliances ranging from 0.0025 to 0.03 L/cm H(2)O. Volumes and pressures were measured using a calibrated screen pneumotachograph and custom software. We tested the Smartvent 7900, Avance, and Aisys anesthesia ventilator systems (GE Healthcare, Madison, WI) and the Apollo anesthesia ventilator (Draeger Medical, Telford, PA). The Smartvent 7900 and Avance ventilators use inspiratory flow sensors to control the volume delivered, whereas the Aisys and Apollo ventilators compensate for the compliance of the circuit. RESULTS: We found that the anesthesia ventilators that use compliance compensation (Aisys and Apollo) accurately delivered both large and small tidal volumes to the airway of the test lung under conditions of normal and low lung compliance during VCV (ranging from 95.5% to 106.2% of the set tidal volume). However, the anesthesia ventilators without compliance compensation were less accurate in delivering the set tidal volume during VCV, particularly at lower volumes and lower lung compliances (ranging from 45.6% to 100.3% of the set tidal volume). CONCLUSIONS: Newer generation anesthesia machine ventilators that compensate for breathing circuit compliance and for fresh gas flow are able to deliver small tidal volumes accurately to the airway under conditions of normal and low lung compliance during volume-controlled ventilation. Accurate VCV may be a useful alternative to PCV, as volume is guaranteed when lung compliance changes, and new strategies such as small volume/lung protective ventilation become possible in the operating room. PMID- 18420851 TI - The prophylactic effect of haloperidol plus dexamethasone on postoperative nausea and vomiting in patients undergoing laparoscopically assisted vaginal hysterectomy. AB - BACKGROUND: Haloperidol, a major tranquilizer, has been found to have a potent antiemetic effect on postoperative nausea and vomiting (PONV), but the prophylactic effect of haloperidol plus dexamethasone on PONV has not been evaluated. We evaluated the prophylactic effect of haloperidol plus dexamethasone to either given alone, placebo or droperidol on PONV in patients undergoing a laparoscopic-assisted vaginal hysterectomy. METHODS: Four hundred adult women (n = 80 in each of five groups) scheduled for a laparoscopic-assisted vaginal hysterectomy were enrolled in a randomized, double-blind, placebo, and positive control study. Fifteen minutes after the induction of anesthesia, patients received an i.v. injection of either saline (group S), droperidol 1.25 mg (group D), haloperidol 2 mg (group H), dexamethasone 5 mg (group Dx), or haloperidol 2 mg plus dexamethasone 5 mg (group H + Dx) to prevent PONV. The occurrence of PONV and medication-related side effects were recorded. RESULTS: The incidences of PONV (0-24 h) in the D (36%), H (37%), Dx (38%), and H + Dx (19%) groups were significantly lower than in the S group (65%; P < 0.05 for each comparison). The H + Dx group had the lowest incidence of PONV (19%; P < 0.05 for each comparison) of the five study groups. No differences were found between the D, H, and Dx groups. Also, no differences were found among the five groups in the side effects of QT prolongation, intensity of postoperative pain, level of sedation, and occurrence of extra-pyramidal symptoms. CONCLUSION: Prophylactic haloperidol 2 mg plus dexamethasone 5 mg produced a greater reduction in the incidence of PONV than did either drug used alone, placebo or droperidol without increasing perioperative adverse outcomes. PMID- 18420852 TI - Haloperidol versus ondansetron for prophylaxis of postoperative nausea and vomiting. AB - BACKGROUND: Haloperidol is effective for postoperative nausea and vomiting prophylaxis, but there are almost no data comparing it to 5-HT(3) antagonists. METHODS: Two hundred forty-four adults were randomized to receive i.v. haloperidol 1 mg or ondansetron 4 mg, during general anesthesia. Nausea, vomiting, need for rescue, sedation, extrapyramidal effects, QTc intervals, and time to postanesthesia care unit discharge were evaluated with a third-party blind design. RESULTS: There was no intergroup difference in any measure of efficacy or toxicity. Haloperidol and ondansetron subjects (78.2% and 76.8%) had complete response. Postoperatively, prolonged QTc occurred in 28.9% and 22.1% (N.S.). CONCLUSIONS: In a mixed surgical population, the efficacy and toxicity of postoperative nausea and vomiting prophylaxis with haloperidol 1 mg was not significantly different from ondansetron 4 mg. PMID- 18420853 TI - Haloperidol plus ondansetron versus ondansetron alone for prophylaxis of postoperative nausea and vomiting. AB - INTRODUCTION: Haloperidol 1 mg and ondansetron 4 mg are equally safe and effective for postoperative nausea and vomiting prophylaxis. We compared the combination to ondansetron alone in a mixed surgical population. METHODS: Two hundred and sixty-eight adults undergoing general anesthesia received 4 mg ondansetron plus 1 mg haloperidol or saline in this randomized, double-blind protocol. Efficacy and safety data were obtained until 480 min after postanesthesia care unit entry. RESULTS: The combination had more complete responders (76.2% vs 59.2%), less nausea, less rescue, and longer time to rescue. Sedation, time to postanesthesia care unit discharge, and QTc prolongation were not different. No subject had dystonia, akathisia, or serious dysrhythmias. CONCLUSIONS: Postoperative nausea and vomiting prophylaxis with both drugs is significantly more effective and longer lasting than ondansetron alone. There is no detectable increase in side effects. PMID- 18420854 TI - Pro: The Food and Drug Administration Black box warning on droperidol is not justified. PMID- 18420855 TI - Con: The black box warning on droperidol should not be removed (but should be clarified!). PMID- 18420856 TI - Risk factors associated with fast-track ineligibility after monitored anesthesia care in ambulatory surgery patients. AB - BACKGROUND: Fast-tracking after ambulatory anesthesia has been advocated as a pathway to improve efficiency and maximize resources without compromising patient safety and satisfaction. Studies reporting successful fast-tracking focus primarily on anesthesia techniques and not on specific patient factors, surgical procedure, or process variables associated with unsuccessful fast-tracking. We performed this retrospective study to implement a process for improving fast tracking, measure change over time, and identify variables associated with patients unable to fast-track successfully after monitored anesthesia care. METHODS: A fast-track protocol for all patients receiving monitored anesthesia care based on the Modified Aldrete Score was instituted. It consisted of written policy changes and weekly review at physician and nursing department meetings for the first month, followed by monthly feedback during a 6-mo intervention period. Data collected for a 3-mo baseline and the consecutive 6-mo intervention period included fast-track status, surgical service and procedure, surgeon and anesthesiology provider, age, gender, ASA status, total time in operating room, and total postoperative time (end of surgery to actual discharge). RESULTS: Three hundred and thirty-two cases were completed during the 3-mo baseline period, and 641 cases were completed during the 6-mo intervention period. Fast-track success rate improved from 23% to 56%, P < 0.001. Independent risk factors for fast-track ineligibility identified by multivariate regression analysis were significant for patients <60 yr-old, ASA III versus I, general surgery versus orthopedics and ophthalmology, month after implementation, and total postoperative time. Total postoperative time was significantly shorter by 64 min in the fast-track group, P < 0.001. CONCLUSION: Fast-track success rate can be improved and sustained over time by education and personnel feedback. We identified risk factors that were significantly associated with fast-track ineligibility. If those factors are found to be associated with fast-track ineligibility in a prospective investigation, they should enable development of multidisciplinary patient and procedure-specific guidelines for fast-tracking. PMID- 18420857 TI - The effects of general anesthesia on the central nervous and cardiovascular system toxicity of local anesthetics. AB - BACKGROUND: Local anesthetic toxicity is often studied experimentally in acutely prepared, anesthetized laboratory animals. We determined the influence of halothane/O(2) anesthesia on cardiovascular and central nervous system (CNS) toxic responses to six amide-type local anesthetics administered i.v.. METHODS: Behavioral, cardiovascular, and pharmacokinetic responses were determined in previously instrumented ewes (approximately 45-50 kg, n = 18), on separate occasions when conscious and anesthetized, to bupivacaine (100 mg), levobupivacaine (125 mg), ropivacaine (150 mg), lidocaine (350 mg), mepivacaine (350 mg), prilocaine (350 mg), and saline (control) infused i.v. over 3 min. RESULTS: The local anesthetics caused convulsions in conscious sheep, but no overt CNS effects in anesthetized sheep. Negative inotropy and slight bradycardia without changes in arterial blood pressure occurred initially in conscious sheep, followed by positive inotropy, tachycardia, and hypertension at the abrupt onset of CNS excitotoxicity, along with widening of QRS complexes. Fatal cardiac arrhythmias occurred in, respectively, 3 of 11, 2 of 12, and 2 of 13 conscious sheep infused with bupivacaine, levobupivacaine, and ropivacaine; in 1 of 9 with prilocaine, electromechanical dissociation (followed by polymorphic ventricular tachycardia) caused death. In anesthetized sheep, cardiovascular depression, preexisting from the general anesthesia, was exacerbated by all local anesthetics, and increased QRS width was prolonged; concurrent blood local anesthetic concentrations were doubled. Nevertheless, all anesthetized animals survived. CONCLUSIONS: General anesthesia produced physiological perturbations, exacerbated local anesthetic-induced cardiovascular depression, and changed the pharmacokinetics of toxic doses of local anesthetics. However, cardiovascular fatalities from local anesthetics occurred only in conscious animals. PMID- 18420858 TI - The effects of general anesthesia on whole body and regional pharmacokinetics of local anesthetics at toxic doses. AB - BACKGROUND: Local anesthetic toxicity is often studied experimentally in anesthetized subjects, but clinical toxicity usually occurs in conscious patients. In this study, we determined the influence of general anesthesia on the pharmacokinetics of six local anesthetics administered i.v. at approximately the highest recommended doses. METHODS: Chronically instrumented ewes (approximately 45-50 kg, n = 18) were infused over 3 min with (base doses as HCl salts) bupivacaine (100 mg), levobupivacaine (125 mg), ropivacaine (150 mg), lidocaine (350 mg), mepivacaine (350 mg), or prilocaine (350 mg), on separate occasions when conscious and halothane anesthetized. Serial arterial, heart, and brain venous blood drug concentrations were measured by achiral/chiral high-performance liquid chromatography, as relevant. Whole body pharmacokinetics were assessed by noncompartmental analysis; heart and brain pharmacokinetics were assessed by mass balance. Drug blood binding, in the absence and presence of halothane, was assessed by equilibrium dialysis in vitro. RESULTS: Blood local anesthetic concentrations were doubled with anesthesia because of decreased whole body distribution and clearance (respectively, to 33% and 52% of values when conscious). Heart and brain net drug uptake were greater under anesthesia, reflecting slower efflux from both regions. Clearances of R-bupivacaine > S bupivacaine and R-prilocaine > S-prilocaine, but, mepivacaine clearance was not enantioselective. Halothane did not influence blood binding of the local anesthetics. CONCLUSIONS: General anesthesia significantly changed whole body and regional pharmacokinetics of each local anesthetic as well as the systemic effects. General anesthesia is thus an important but frequently overlooked factor in studies of local anesthetic toxicity. PMID- 18420859 TI - Propranolol increases the threshold for lidocaine-induced convulsions in awake rats: a direct effect on the brain. AB - BACKGROUND: Propranolol is a beta-adrenoceptor antagonist used clinically. Local anesthetics are used for controlling pain, whereas propranolol is concomitantly given to treat hypertension and tachycardia. However, there are few studies examining the effects of propranolol on the toxicity of local anesthetics. We investigated the effect of propranolol on lidocaine-induced convulsions in awake, spontaneously breathing rats. METHODS: Male Sprague-Dawley rats were randomly divided into six groups (n = 8, each group). Rats were pretreated with intracerebroventricular saline (cerebroventricle-control: CV-C group), 10 or 30 microg/kg of (S)-(-)-propranolol (propranolol) (cerebroventricle-small dose: CV-S and cerebroventricle-large dose: CV-L groups, respectively) or i.v. saline (IV control: IV-C group), 1 or 3 mg/kg of propranolol (IV-small dose: IV-S and IV large dose: IV-L groups, respectively). Three minutes later, lidocaine was administered i.v. at 4 mg x kg(-1) x min(-1) until tonic-clonic convulsions occurred. RESULTS: The convulsive dose of lidocaine in the CV-L group was significantly larger than that in the CV-C group (30.6 +/- 5.1 vs 23.5 +/- 2.2 mg/kg, respectively, P = 0.008). Plasma concentrations of total and protein unbound lidocaine, concentrations of lidocaine in the brain at the onset of convulsions were also significantly higher in the CV-L group than those in the CV C group (36.1 +/- 4.8 vs 26.0 +/- 3.8 microg/mL, 22.5 +/- 3.5 vs 13.7 +/- 2.6 microg/mL, 82.7 +/- 7.1 vs 57.3 +/- 5.7 microg/g, P < 0.001 for all). The convulsive dose, plasma concentrations of total and protein-unbound lidocaine, and brain lidocaine in the IV-L group were also significantly larger than those in IV-C group and comparable with those in the CV-L group. The plasma concentration of propranolol before starting an infusion of lidocaine in the IV-L group was approximately 60-fold higher than that in the CV-L group (554.7 +/- 104.6 and 9.3 +/- 6.7 ng/mL, respectively). CONCLUSIONS: Propranolol increased the threshold for lidocaine-induced convulsions by directly acting on the brain. PMID- 18420860 TI - In vitro, inhibition of mitogen-activated protein kinase pathways protects against bupivacaine- and ropivacaine-induced neurotoxicity. AB - BACKGROUND: Animal models show us that specific activation of the p38 mitogen activated protein kinase (MAPK) may be a pivotal step in lidocaine neurotoxicity, but this has not been investigated in the case of two very widely used local anesthetics, bupivacaine and ropivacaine. We investigated the hypotheses that these drugs (A) are less neurotoxic than the prototype local anesthetic, lidocaine (B) are selectively toxic for subcategories of dorsal root ganglion neurons and (C) induce activation of either p38 MAPK or related enzymes, such as the c-jun terminal N-kinase (JNK) and extracellular signal-regulated kinase (ERK). METHODS: We incubated primary sensory neuron cultures with doses of lidocaine, bupivacaine, and ropivacaine equipotent at blocking sodium currents. Next, we sought to determine potential selectivity of bupivacaine and ropivacaine toxicity on neuron categories defined by immunohistochemical staining, or size. Subsequently, the involvement of p38 MAPK, JNK, and ERK was tested using enzyme linked immunosorbent assays. Finally, the relevance of MAPK pathways in bupivacaine- and ropivacaine-induced neurotoxicity was determined by selectively inhibiting activity of p38 MAPK, JNK, and ERK. RESULTS: We found that the neurotoxic potency of bupivacaine and ropivacaine is dose-dependent and similar in vitro, but is not selective for any of the investigated subgroups of neurons. Neurotoxicity of bupivacaine and ropivacaine was mediated, at least in part, by MAPKs. Specifically, we demonstrated the relevance of both p38 MAPK and JNK pathways for the neurotoxicity of bupivacaine and characterized the involvement of the p38 MAPK pathway in the neurotoxicity of ropivacaine. CONCLUSIONS: Given equipotent doses, the neurotoxic potential of lidocaine does not appear to be significantly different from that of bupivacaine and ropivacaine in vitro. Moreover, bupivacaine and ropivacaine do not exert their neurotoxicity differently on specific subsets of dorsal root ganglion neurons. Their neurotoxic effects are brought about through the activation of specific MAPKs; the specific pharmacologic inhibition of these kinases attenuates toxicity in vitro. PMID- 18420861 TI - Lidocaine increases intracellular sodium concentration through a Na+-H+ exchanger in an identified Lymnaea neuron. AB - BACKGROUND: The intracellular sodium concentration ([Na(+)]in) is related to neuron excitability. For [Na(+)]in, a Na(+)-H(+) exchanger plays an important role, which is affected by intracellular pH ([pH]in). However, the effect of lidocaine on [pH]in and a Na(+)-H(+) exchanger is unclear. We used neuron from Lymnaea stagnalis to determine how lidocaine affects [pH]in, Na(+)-H(+) exchanger, and [Na(+)]in. METHODS: Intracellular sodium imaging by sodium-binding benzofuran isophthalate and intracellular pH imaging by 2',7'-bis(2-carboxyethyl) 5(6)-carboxyfluorescein were used to measure [Na(+)]in and [pH]in. Measurements for [Na(+)]in were made in normal, Na(+) free saline, with modified extracellular pH, and a Na(+)-H(+) exchanger antagonist [(5-N-ethyl-N-isopropyl amiloride, N methylisopropylamiloride, and 5-(N,N-hexamethylene)-amiloride) pretreatment trials. Furthermore, [Na(+)]in and [pH]in were recorded simultaneously. From 0.1 to 10 mM, lidocaine, mepivacaine, bupivacaine, prilocaine, and QX-314 were evaluated. RESULTS: Lidocaine, mepivacaine, and prilocaine increased the [Na(+)]in in a dose-dependent manner. In contrast, QX-314 did not change the [Na(+)]in at each dose. In the Na(+) free saline or in the presence of each Na(+) H(+) exchanger antagonist, lidocaine failed to increase [Na(+)]in. Lidocaine, mepivacaine, and prilocaine induced a significant decrease in [pH]in below baseline with an increase in [Na(+)]in. In contrast, QX-314 did not change the [pH]in. These results demonstrated that lidocaine increases [Na(+)]in through Na(+)-H(+) exchanger activated by intracellular acidification, which is induced by the proton trapping of lidocaine. This [Na(+)]in increase and [pH]in change induces cell toxicity. CONCLUSION: Lidocaine increases the [Na(+)] through a Na(+)-H(+) exchanger by proton trapping. PMID- 18420862 TI - Desflurane preconditioning inhibits endothelial nuclear factor-kappa-B activation by targeting the proximal end of tumor necrosis factor-alpha signaling. AB - BACKGROUND: Volatile anesthetics interfere with inflammatory cytokine production and expression of adhesion molecules which are critical for ischemia reperfusion induced injury. Nuclear factor (NF)-kappaB has been reported to be suppressed in this process, but the detailed molecular mechanism is still unclear. METHODS: In this study, ECV304 (a human umbilical vein endothelial cell line) was preconditioned with 30 min desflurane (1 minimal alveolar concentration), after 15 min washout, 30 min anoxia, and 60 min reoxygenation was performed. ECV304 was finally stimulated with tumor necrosis factor (TNF)-alpha (10 ng/mL). Control groups, which were not preconditioned and/or not stimulated, were also included in the protocol. IkappaB-alpha, phospho-IkappaB-alpha, phospho-IkappaB kinase (IKKalpha)/IKKbeta, and phospho-p38 were detected by Western blotting. The nuclear NF-kappaB p65 subunit was measured by subcellular fractionation and Western blotting. The surface expression of TNF-R1 was measured by flow cytometry. Receptor-associated signaling adaptors, e.g., TNF receptor-associated factor 2 (TRAF2) and IKK-alpha, were evaluated by immunoprecipitation by TNF-R1 antibody and subsequent Western blotting. RESULTS: Desflurane preconditioning inhibits IkappaB-alpha phosphorylation, degradation, and p65 nuclear localization. Desflurane also affects p38 phosphorylation, which is needed for optimal inflammatory response. The phosphorylation of IKKalpha/IKKbeta was suppressed by preconditioning while the surface abundance of TNF-R1 was not affected. The association of TRAF2 and IKK-alpha with TNF-R1 was compromised by desflurane. CONCLUSIONS: Our results suggest that the molecular target of desflurane in the NF-kappaB pathway is upstream of IKK activation. The abundance of TNF-R1 on the cell membrane is not affected by anesthetic preconditioning. We suggest that desflurane preconditioning targets the proximal end of TNF-alpha signaling. PMID- 18420863 TI - Cardiac output measurement in patients undergoing liver transplantation: pulmonary artery catheter versus uncalibrated arterial pressure waveform analysis. AB - BACKGROUND: Cardiac output (CO) and invasive hemodynamic measurements are useful during liver transplantation. The pulmonary artery catheter (PAC) is commonly used for these patients, despite the potential complications. Recently, a less invasive device (Vigileo/FloTrac) became available, which estimates CO using arterial pressure waveform analysis without external calibration. In this study, we compared CO obtained with a PAC using automatic thermodilution, instantaneous CO stat-mode (ICO(SM)), and CO obtained with the new device, arterial pressure waveform analysis (APCO) in patients undergoing liver transplantation. METHODS: Twenty sets of simultaneous measurements of APCO and ICO(SM) were determined in sedated and mechanically ventilated patients undergoing liver transplantation. Time points were as follows: after PAC insertion (T1-3), after portal clamping (T4-6), during anhepathy (T7-9), after graft reperfusion (T10-15), and in the postoperative period in the intensive care unit (T15-20). RESULTS: We enrolled 20 patients and 400 measurements were obtained. No data were rejected. Bias between ICO(SM) and APCO was 0.8 L/min, 95% limits of agreement were -1.8 to 3.5 L/min. The percentage error was 43%. Bias between ICO(SM) and APCO was correlated with systemic vascular resistance [r(2) = 0.55, P < 0.0001, y = 15.8-2.2 ln(x)] and subgroup analysis revealed an increase in the bias and in the percentage error in patients with low systemic vascular resistance (Child-Pugh grade B and C patients). There was no difference between the different surgical periods. CONCLUSIONS: Our results suggest that Vigileo/FloTrac CO monitoring data do not agree well with those of automatic thermodilution in patients undergoing liver transplantation, especially in Child-Pugh grade B and C patients with low systemic vascular resistance. PMID- 18420864 TI - Changing patterns in anesthetic fresh gas flow rates over 5 years in a teaching hospital. AB - BACKGROUND: Reducing anesthetic fresh gas flows can reduce volatile anesthetic consumption without affecting drug delivery to the patient. Delivery systems with electronic flow transducers permit the simple and accurate collection of fresh gas flow information. In a 2001 audit of fresh gas flow, we found little response to interventions designed to foster more efficient use of fresh gas. We compared current practice with our earlier results. METHODS: Flow data were collected in areas with a mix of general and acute surgery in March and November 2001, and again during 2006, by recording directly from the Datex ADU to a computer every 10 s. We extracted the distribution of flow rates when a volatile anesthetic was being administered. Data collection in March 2001 and 2006 was not advertised. RESULTS: In 2001, the mean flow rates were 1.95 and 2.1 L/min with a median flow of 1.5 L/min. In 2006, the mean was 1.27 and the median in the range 0.5-1.0 L/min. Isoflurane use decreased from 47% in 2001 to 4% in 2006. CONCLUSIONS: Fresh gas flows used in our department have decreased by 35% over 4 years. Although the absolute change in flow rate is not large, this represents potential annual savings of more than $US130,000. This occurred without specific initiatives, suggesting an evolution in practice towards lower fresh gas flow. Improvements in equipment and monitoring, including a locally developed system, which displays forward predictions of end-tidal and effect-site vapor concentrations, may be factors in this change. PMID- 18420865 TI - The impact of different step changes of inspiratory fraction of oxygen on functional residual capacity measurements using the oxygen washout technique in ventilated patients. AB - BACKGROUND: Functional residual capacity (FRC) measurements may help to guide respiratory therapy. Using the oxygen washout technique, FRC can be assessed at bedside during spontaneous breathing. High repeatability, crucial for monitoring, has not been shown in ventilated patients. A large step change of inspiratory fraction of oxygen (FiO(2)) (DeltaFiO(2)) may impede the clinical use in patients ventilated with high FiO(2). We investigated the repeatability of FRC measurements and the impact of different DeltaFiO(2) on this repeatability. METHODS: The LUFU system (Draeger Medical, Luebeck, Germany) estimates FRC by oxygen washout, a variant of multiple-breath-nitrogen-washout during a fast DeltaFiO(2). In 20 postoperative cardiac surgery patients, FRC was measured in duplicate using DeltaFiO(2) of 0.1, 0.2, and 0.6. RESULTS: There were no differences between repeated measurements of FRC, neither using a DeltaFiO(2) of 0.1, 0.2 nor 0.6(Delta0.1: 2.62 L +/- 0.58, 2.62 L +/- 0.59, P = 0.995; Delta0.2: 2.70 L +/- 0.59, 2.66 L +/- 0.56, P = 0.258; Delta0.6: 2.61 L +/- 0.58, 2.59 L +/ 0.58, P = 0,639). Coefficients of variation were 6.6%, 5.6%, and 6.6%, respectively. CONCLUSIONS: FRC can be measured in ventilated patients using the oxygen washout technique with a clinically acceptable repeatability. Repeatability is not significantly influenced whether using a DeltaFiO(2) of 0.1, 0.2, or 0.6. PMID- 18420866 TI - Poor visualization during direct laryngoscopy and high upper lip bite test score are predictors of difficult intubation with the GlideScope videolaryngoscope. AB - BACKGROUND: The GlideScope videolaryngoscope allows equal or superior glottic visualization compared with direct laryngoscopy, but predictive features for difficult GlideScope intubation have not been identified. We undertook this prospective study to identify patient characteristics associated with difficult GlideScope intubation. METHODS: Demographic and morphometric factors were recorded preoperatively for 400 patients undergoing anesthesia with endotracheal intubation. After induction, direct laryngoscopy was performed in all patients to assess the Cormack and Lehane grade of glottic visualization followed by GlideScope intubation. The number of attempts and time needed for intubation were recorded. Univariate and multivariate analyses were performed to identify the characteristics associated with difficult GlideScope intubation. RESULTS: Intubation required 1, 2, and 3 attempts in 342, 48, and 9 participants, respectively, with one failure. Mean time for intubation was 21 +/- 14 s. After univariate analysis, the following characteristics were significantly correlated (P < 0.05) with longer time to intubate and/or multiple attempts: older age, male sex, history of snoring, high Mallampati class, small mouth opening, short sternothyroid and manubriomental distances, large neck circumference, high upper lip bite test score, and high Cormack and Lehane grade during direct laryngoscopy. However, after introducing these variables in nominal logistic and proportional hazard multiple regression models, only high Cormack and Lehane grade during direct laryngoscopy, high upper lip bite test score, and short sternothyroid distance were significantly associated with multiple attempts or lengthier intubations. CONCLUSION: Despite a high success rate, intubation with the GlideScope is likely to be more challenging in patients with high Cormack and Lehane grade during direct laryngoscopy, high upper lip bite test score, or short sternothyroid distance. PMID- 18420867 TI - Fiberoptic intubation through an I-gel supraglottic airway in two patients with predicted difficult airway and intellectual disability. AB - We describe successful fiberoptic-guided tracheal intubation through the novel supraglottic "I-gel" airway in two uncooperative adult patients with genetic syndromes, learning disability, and predicted difficult airway, scheduled for complex dental treatment under general anesthesia. The I-gel maintained the airway immediately after induction, allowing oxygenation and ventilation. Location of the laryngeal inlet was successful on the first attempt with a fiberscope, and the tracheal tube was inserted into the trachea over the endoscope without complication in both patients. This report suggests another option for management of predicted difficult airways. PMID- 18420868 TI - Effects of a suction laryngoscope in a model with simulated severe airway hemorrhage. AB - In severe airway hemorrhage, simultaneous suction and laryngoscopy may render intubation difficult. We built a suction laryngoscope that consists of an adjustable stainless steel-guide tube fixed at the lingual surface of a standard Macintosh laryngoscope blade. Via this steel-guide tube, a large suction catheter can be inserted and positioned exactly to suction pharyngeal blood or vomitus, rendering simultaneous suctioning and laryngoscopy possible. In contrast to previous suction laryngoscopes, our suction catheter has a large lumen, which enables fast suctioning and exact placement by adjusting the steel-guide tube. To assess whether our suction laryngoscope could provide better intubation conditions in comparison to a standard Macintosh laryngoscope in a bleeding airway scenario, 44 medical students intubated a manikin with severe simulated airway hemorrhage using our suction laryngoscope and a standard Macintosh laryngoscope in random order. There was no significant difference in time needed for intubation when using the suction versus the Macintosh laryngoscope (mean +/- SD: 43 +/- 13 vs 52 +/- 31 s; P = 0.07), but the number of esophageal intubations was significantly lower when using the suction laryngoscope [6 of 44 (13.6%) vs 19 of 44 (43.2%); P = 0.004]. In conclusion, when compared with a standard Macintosh laryngoscope, using a suction laryngoscope did not result in more rapid intubation, but significantly decreased the likelihood of esophageal intubations. PMID- 18420869 TI - The development and validation of a dynamic model to account for the progress of labor in the assessment of pain. AB - BACKGROUND: Labor pain is often described as the worst pain in a woman's life, but the experience is highly variable. Although many factors have been linked to labor pain, it has been difficult to assess the individual effects of these factors because labor is a dynamic process and pain intensity changes over the course of labor. Previous studies have used average pain scores. The aim of this study was to develop and validate a model that would allow for the statistical analysis of factors that affect pain throughout labor. METHODS: We conducted this study with a retrospective database drawn from the medical records of 200 consecutive nulliparous parturients who delivered at New York Presbyterian Hospital between October 2006 and January 2007. Numerical rating scale scores for pain with contractions (0-10 scale), cervical dilation, and oxytocin use before analgesia request were recorded. Nonlinear effects modeling with a sigmoid equation was used to describe the relationship between reported pain and cervical dilation. The modeling technique was developed with data from 91 parturients and validated with an independent set of 95 parturients (all parturients with pain scores more than zero). The resulting model was used to analyze the effect of a sample covariate, oxytocin administration, on reported pain in the entire data set. RESULTS: The model derived from our training set was predictive of the data from our validation set (P < 0.001). Predicted pain scores were on average two numerical rating scale points above or below measured pain scores. Analyzing oxytocin as a covariant showed that women treated with oxytocin reported 48% more pain at the start of labor but did not have a significantly more rapid increase in pain or higher maximal pain when compared with women not treated with oxytocin. Women treated with oxytocin had slower early labor and more rapid late labor. CONCLUSION: We have developed and validated a model for describing pain over the course of labor. Our model is suited to the statistical analysis of covariance and could potentially be used to compare the effects of covariants on labor pain and the rate of change of pain. PMID- 18420870 TI - The time-dependent effects of midazolam on regional cerebral glucose metabolism in rats. AB - BACKGROUND: Midazolam has hypnotic and sedative activities, which may be mediated by different neuronal structures. We investigated the time course effect of a hypnotic dose of midazolam on conscious motor behavior and on patterns of brain metabolism. METHODS: Loss of nociceptive reflexes and impairment of spontaneous locomotor activity were used as indices for the hypnotic and sedative effects of midazolam, and the regional cerebral metabolic rates for glucose (rCMRglc) were used as indices of neuronal effects of midazolam. Locomotor activity was measured with a monitor and rCMRglc were measured with the quantitative autoradiographic [(14)C]2-deoxyglucose procedure in 62 brain regions of Fischer-344 rats at 2, 30, 60, 120, and 180 min after i.v. administration of saline or midazolam 5 mg/kg. RESULTS: After midazolam administration, rats were anesthetized at 2 min, awake but severely impaired at 30 min and slowly recovering motor activity thereafter. Anesthesia was associated with widespread rCMRglc decreases (59 areas affected, 38% mean decrease). Recovery of consciousness was associated with normalizing rCMRglc in visual, auditory, and somatosensory cortices and in the locus coeruleus (47 regions affected, 31% decrease). Recovery of motor activity was paralleled by slow rCMRglc normalization in the frontal motor, limbic, and thalamic regions (at 60, 120, and 180 min 31, 17, 4 areas affected, 26, 20, and 15% decreases from control values). CONCLUSIONS: Whereas the hypnotic effects of midazolam may result from inhibition of brain structures involved in arousal and sensory processing, its sedative effects may result from inhibition of subcortical motor and limbic regions. PMID- 18420871 TI - Active research fields in anesthesia: a document co-citation analysis of the anesthetic literature. AB - BACKGROUND: The expansion of science has resulted in an increased information flow and in an exponentially growing number of connections between knowledge in different research fields. In this study, we used methods of scientometric analysis to obtain a conceptual network that forms the structure of active scientific research fields in anesthesia. METHODS: We extracted from the Web of Science (Institute for Scientific Information) all original articles (n = 3275) including their references (n = 79,972) that appeared in 2003 in all 23 journals listed in the Institute for Scientific Information Journal Citation Reports under the subject heading "Anesthesiology." After identification of highly cited references (> or = 5), pairs of co-cited references were created and grouped into uniformly structured clusters of documents using a single linkage and variable level clustering method. In addition, for each such cluster of documents, we identified corresponding front papers published in 2003, each of which co-cited at least two documents of the cluster core. Active anesthetic research fields were then named by examining the titles of the documents in both the established clusters and in their corresponding front papers. These research fields were sorted according to the proportion of recent documents in their cluster core (immediacy index) and were further analyzed. RESULTS: Forty-six current anesthetic research fields were identified. The research field named "ProSeal laryngeal mask airway" showed the highest immediacy index (100%) whereas the research fields "Experimental models of neuropathic pain" and "Volatile anesthetic-induced cardioprotection" exhibited the highest level of co-citation strength (level 9). The research field with the largest cluster core, containing 12 homogeneous papers, was "Postoperative nausea and vomiting." The journal Anesthesia & Analgesia published most front papers while Anesthesiology published most of the fundamental documents used as references in the front papers. CONCLUSIONS: Using co-citation analysis, we identified distinct homogenous clusters of highly cited documents representing 46 active current anesthetic research fields and determined multiple nets of knowledge among them. PMID- 18420872 TI - Predictive factors of severe postoperative pain in the postanesthesia care unit. AB - BACKGROUND: I.v. morphine titration (IMT) is widely used in the postanesthesia care unit to achieve pain relief. Numerous factors contribute to variability in postoperative pain or morphine consumption. We analyzed prospectively the pre- and intraoperative predictive factors of severe postoperative pain defined as a dose of IMT >0.15 mg/kg or a failure of IMT. METHODS: We assessed the role of preoperative information about pain, medical treatments, and intraoperative events and their role on postoperative pain. After IMT, patients were divided into two groups: severe pain (SP) and nonsevere pain. Data are expressed as mean +/- SD. RESULTS: Three hundred forty-two patients were included in the study: 200 (58%) in the nonsevere pain group and 142 (42%) in the SP group. Using a univariate analysis, there was no significant difference between groups related to medical or surgical history except for more frequent preoperative treatments in the SP group (P < 0.05). Duration of the surgical procedure and anesthesia were longer in the SP group (P < 0.001). The dose of sufentanil and visual analog scale scores before and at the end of IMT were higher in the SP group (P < 0.001). Using a multivariate analysis, a high dose of intraoperative opioid (sufentanil dose >0.6 microg/kg) (Odds ratio = 2.68, P < 0.001), a general anesthetic procedure (Odd ratio = 3.96, P = 0.03), and the use of preoperative analgesic drugs (Odds ratio = 1.91, P < 0.01) were independent factors associated with severe postoperative pain. CONCLUSION: A higher intraoperative dose of sufentanil, general anesthesia, and preoperative treatment with analgesics were significantly associated with severe postoperative pain. PMID- 18420873 TI - Sonographic estimation of needle depth for cervical epidural blocks. AB - BACKGROUND: Cervical epidural steroid injections are often used to treat acute and chronic pain syndromes involving the face, neck, and upper extremities. Ultrasound has evolved as a valuable tool for performing neuraxial blocks, providing useful prepuncture information on the structure. Our goal was to evaluate the accuracy and precision of ultrasound by comparing skin to dura distance from ultrasound with the actual skin to epidural depth. METHODS: We enrolled 50 patients undergoing cervical epidural blocks at the pain clinic. Ultrasound images with transverse and longitudinal median views of the C6/7 area were taken. The epidural needle was inserted, reproducing the direction of the ultrasound beam on the longitudinal median view. Measured distances from skin to dura on each ultrasound view were compared with the actual needle depth. Additionally, we examined ultrasound visibility, the number of puncture attempts, and any complications related to the procedure. RESULTS: Concordance correlation coefficients between the measured distances on ultrasound and actual needle depth were 0.9272 and 0.9268 on transverse and longitudinal median view, respectively. The cervical epidural block was successfully performed on 48 patients (96%). There were two incidents (4%) of dural puncture. No bloody taps, postprocedure complications, or hemodynamic instability related to cervical epidural blocks occurred. CONCLUSIONS: Ultrasound provides very accurate information on the skin to dura distance for epidural blocks in the cervical spine. Knowledge of skin to dura distance and a preview of spinal anatomy before puncture can more safely identify the epidural space. PMID- 18420874 TI - Intraarticular injection of magnesium sulphate and/or bupivacaine for postoperative analgesia after arthroscopic knee surgery. AB - BACKGROUND: Intraarticular bupivacaine is often used for prevention of pain after arthroscopic knee surgery. Intraarticular magnesium, a N-methyl-D-aspartate receptor blocker, would be of particular interest in either producing postoperative analgesia or enhancing the analgesic effect of intraarticular bupivacaine. We designed this study to determine whether intraarticular magnesium sulfate or bupivacaine results in a decrease in visual analog scale (VAS) score followed by a decrease in analgesic requirement and whether their combination would provide more reduction in VAS, and subsequently less analgesic requirement, than either drug alone. METHODS: One-hundred and eight patients undergoing arthroscopic meniscectomy were randomized blindly into one of four parallel groups. The saline placebo group (group S) received 20 mL of isotonic saline, and the magnesium group (group M) received 20 mL of isotonic saline containing 1 g magnesium sulfate. The bupivacaine group (group B) received 0.25% (20 mL) bupivacaine, whereas the magnesium with bupivacaine group (group MB) received bupivacaine 0.25% and 1 g of magnesium sulfate in 20 mL. The postoperative analgesia was assessed using VAS recorded after surgery at 1, 2, 4, 6, 8, 12, and 24 h. Patients were evaluated at rest and with movement. RESULTS: Group MB had a significantly reduced VAS both at rest and on movement, a significantly increased time to first postoperative analgesic request, as well as significantly reduced total analgesic requirement than other groups. CONCLUSION: Magnesium combined with bupivacaine produces a reduction in postoperative pain when given intraarticularly in comparison to either bupivacaine or magnesium alone, or to saline placebo. PMID- 18420875 TI - Combined ultrasound and neurostimulation guidance for popliteal sciatic nerve block: a prospective, randomized comparison with neurostimulation alone. AB - BACKGROUND: Ultrasound imaging, an effective tool to localize peripheral nerves, may facilitate block performance. However, its usefulness during popliteal sciatic nerve block has not been assessed. METHODS: In this prospective, randomized, patient-blinded study, we compared the block time (as the primary end point) of a popliteal sciatic nerve block with double-injection performed using anatomical landmarks and neurostimulation (NS group; n = 30) versus combined ultrasound and neurostimulation guidance (US-NS group; n = 30). Each block procedure was performed by a single operator. Correct needle placement was defined by a minimal stimulating current < or = 0.5 mA, or, in the US-NS group, by mobilization of the nerve by the needle shaft even if the minimal stimulating current > 0.5 mA. Ten milliliter levobupivacaine 0.5% was administered separately on the tibial and common peroneal nerves without needle adjustment to improve the spread of anesthetic in the US-NS group. All procedures were video-recorded, and a maximum of 7 min was allowed to perform the block. Successful block was defined as complete loss of cold sensation in the sciatic distribution and an inability to perform a plantar and dorsal flexion of the foot at 30 min. RESULTS: Five patients in the NS group and three in US-NS group were excluded from the study for prolonged procedure. Block time was not significantly different between groups. The number of needle passes was lower only for the detection of the first nerve in the US-NS group (1 [1-2] vs 2 [1-6]; P < 0.01). A greater success rate was observed at 30 min in the US-NS group (65% vs 16%; P < 0.001). CONCLUSIONS: Combined ultrasound and neurostimulation guidance does not decrease block time but increases the success rate of popliteal sciatic nerve block observed at 30 min. PMID- 18420876 TI - Postoperative analgesic efficacy of peripheral levobupivacaine and ropivacaine: a prospective, randomized double-blind trial in patients after total knee arthroplasty. AB - BACKGROUND: Several previous trials have characterized the intra- and postoperative effects of the recently introduced local anesthetics, levobupivacaine and ropivacaine, for a variety of continuous peripheral nerve blocks. METHODS: We compared the analgesic efficacy of levobupivacaine 0.125% versus ropivacaine 0.2% via patient-controlled femoral nerve analgesia after total knee arthroplasty. In a double-blind, randomized, prospective design, 60 patients received femoral infusion with either substance. We analyzed postoperative local anesthetic consumption, pain scores, motor block, and opioid requirements over 72 h. RESULTS: Pain scores, motor block incidence, and opioid requirements were low and not different between the groups. Ropivacaine consumption in milligrams was 67% higher than that of levobupivacaine. CONCLUSIONS: Both levobupivacaine 0.125% and ropivacaine 0.2% provide similar analgesia after total knee arthroplasty with the latter being less potent. PMID- 18420877 TI - Combined general/epidural anesthesia (ropivacaine 0.375%) versus general anesthesia for upper abdominal surgery. AB - BACKGROUND: We designed this study to investigate the effect of 0.375% ropivacaine on stress hormone responses and anesthetic requirements in combined epidural/general anesthesia for nephrectomy. METHODS: Thirty-two adults, ASA physical status I-II, undergoing nephrectomy lasting about 2 h for renal carcinoma were randomly assigned to one of two groups: epidural (saline)/general anesthesia (group C) and epidural (0.375% ropivacaine)/general (group R) anesthesia. Induction of anesthesia was performed with target-controlled infusion sufentanil and propofol titrated to achieve bispectral index score (define) between 40 and 50. Endotracheal intubation was facilitated by the target controlled infusion administration of vecuronium. After intubation, anesthesia was maintained with propofol infusion titrated to maintain bispectral index between 40 and 50. The target concentration of sufentanil was titrated according to arterial blood pressure and heart rate. Plasma samples were taken immediately before, 2 min after tracheal intubation, immediately after incision, immediately after the initiation of celiac exploration, 1 h after operation, and immediately after tracheal extubation for measurements of epinephrine, glucose, and cortisol in both groups. RESULTS: The relative changes of plasma concentrations of glucoses and cortisol were higher at 1 h after operation and immediately after extubation in group C when compared with group R. The plasma concentrations of epinephrine in both groups were similar. The requirements of propofol for induction and maintenance of anesthesia were not statistically different in the groups, but the requirement of sufentanil during maintenance was significant reduced in group R (P < 0.05). CONCLUSIONS: Epidural ropivacaine 0.375% suppressed stress hormone responses and sufentanil requirements perioperatively during maintenance of anesthesia for nephrectomy. PMID- 18420878 TI - A comparison of the combination of epinephrine and vasopressin with lipid emulsion in a porcine model of asphyxial cardiac arrest after intravenous injection of bupivacaine. AB - BACKGROUND: In a porcine model, we compared the effect of the combination of vasopressin/epinephrine with that of a lipid emulsion on survival after bupivacaine-induced cardiac arrest. METHODS: After administration of 5 mg/kg of a 0.5% bupivacaine solution i.v., ventilation was interrupted for 2 +/- 0.5 (mean +/- SD) min until asystole occurred. Cardiopulmonary resuscitation (CPR) was initiated after 1 min of untreated cardiac arrest. After 2 min of CPR, 10 animals received, every 5 min, either vasopressin combined with epinephrine or 4 mL/kg of a 20% lipid emulsion. Three minutes after each drug administration, up to three countershocks (4, 4, and 6 J/kg) were administered; all subsequent shocks with 6 J/kg. Blood for determination of the plasma bupivacaine concentration was drawn throughout the experiment. RESULTS: In the vasopressor group, all five pigs survived, whereas none of five pigs in the lipid group had restoration of spontaneous circulation (P < 0.01). There was no significant difference between groups in the plasma concentration of total bupivacaine. CONCLUSION: In this model of a bupivacaine-induced cardiac arrest, the vasopressor combination of vasopressin and epinephrine compared with lipid emulsion resulted in higher coronary perfusion pressure during CPR and survival rates. PMID- 18420879 TI - Successful resuscitation after ropivacaine and lidocaine-induced ventricular arrhythmia following posterior lumbar plexus block in a child. AB - We report the case of a 13-yr-old girl scheduled for knee surgery under general anesthesia and posterior lumbar plexus block. A ventricular arrhythmia developed 15 min after local anesthetic injection. A 20% lipid emulsion was successful in converting the ventricular arrhythmia to a sinus rhythm. This is consistent with previous reports suggesting that lipid emulsion is an effective emergency treatment of local anesthetic toxicity. We recommend the immediate availability of lipid emulsion along with other emergency therapeutics in operating rooms where local anesthetics are used. PMID- 18420880 TI - Reversal of central nervous system and cardiac toxicity after local anesthetic intoxication by lipid emulsion injection. AB - A 91-yr-old man (57 kg, 156 cm, ASA III) received an infraclavicular brachial plexus block for surgery of bursitis of the olecranon. Twenty minutes after infraclavicular injection of 30 mL of mepivacaine 1% (Scandicain) and 5 min after supplementation of 10 mL of prilocaine 1% (Xylonest) using an axillary approach, the patient complained of agitation and dizziness and became unresponsive to verbal commands. In addition, supraventricular extrasystole with bigeminy occurred. Local anesthetic toxicity was suspected and a dose of 200 mL of a 20% lipid emulsion was infused. Symptoms of central nervous system and cardiac toxicity disappeared within 5 and 15 min after the first lipid injection, respectively. Plasma concentrations of local anesthetics were determined before, 20, and 40 min after lipid infusion and were 4.08, 2.30, and 1.73 microg/mL for mepivacaine and 0.92, 0.35, and 0.24 microg/mL for prilocaine. These concentrations are below previously reported thresholds of toxicity above 5 microg/mL for both local anesthetics. Signs of toxicity resolved and the patient underwent the scheduled surgical procedure uneventfully under brachial plexus blockade. PMID- 18420881 TI - Intravenous lipid infusion in the successful resuscitation of local anesthetic induced cardiovascular collapse after supraclavicular brachial plexus block. AB - We describe a case of successful resuscitation with an i.v. lipid infusion of local anesthetic-induced cardiovascular toxicity after supraclavicular brachial plexus block with mepivacaine and bupivacaine. Lipid therapy was initiated after 10 min of unsuccessful resuscitation and resulted in restoration of cardiovascular activity and hemodynamic stability. This case illustrates the utility of i.v. lipid therapy in the treatment of local anesthetic toxicity. PMID- 18420882 TI - Simulation education in anesthesia training: a case report of successful resuscitation of bupivacaine-induced cardiac arrest linked to recent simulation training. AB - Simulation training is rapidly becoming an integral element of the education curriculum of anesthesia residency programs. We report a case of successful resuscitation of bupivacaine-induced cardiac arrest treated with i.v. lipid emulsion by providers who had recently participated in simulation training involving a scenario nearly identical to this case. Upon debriefing, it was determined that the previous training influenced execution of the following steps: rapid problem recognition, prompt initiation of specific therapy in the setting of supportive advanced cardiac life support measures, and coordinated team efforts. Although the true cause of efficient resuscitation and ultimate recovery cannot be proven, the efficiency of the resuscitation process, including timely administration of lipid emulsion, is evidence that simulation may be useful for training providers to manage rare emergencies. PMID- 18420884 TI - Electroencephalogram-based indexes indicate the drug effect on the brain, not the (un)consciousness itself. PMID- 18420885 TI - Awareness despite low spectral entropy values. PMID- 18420886 TI - FDA response to droperidol black box warning editorials. PMID- 18420887 TI - The use of lidocaine for spinal anesthesia. PMID- 18420888 TI - Education in anesthesiology should be evidenced-based. PMID- 18420889 TI - Which hemostatic changes determine clinical outcome? PMID- 18420890 TI - The assessment of publication quality. PMID- 18420891 TI - Tension pneumothorax during one-lung ventilation. PMID- 18420892 TI - Early treatment of tetanus-induced trismus with botulinum toxin A. PMID- 18420893 TI - Use of droperidol for ambulatory eye surgery in a patient with Huntington's chorea. PMID- 18420894 TI - Gastric rupture and pneumoperitoneum caused by oxygen insufflation via a fiberoptic bronchoscope. PMID- 18420895 TI - Aprotinin and urinary neutrophil gelatinase-associated lipocalin after cardiac surgery. PMID- 18420897 TI - Quasi-particle properties from tunneling in the v = 5/2 fractional quantum Hall state. AB - Quasi-particles with fractional charge and statistics, as well as modified Coulomb interactions, exist in a two-dimensional electron system in the fractional quantum Hall (FQH) regime. Theoretical models of the FQH state at filling fraction v = 5/2 make the further prediction that the wave function can encode the interchange of two quasi-particles, making this state relevant for topological quantum computing. We show that bias-dependent tunneling across a narrow constriction at v = 5/2 exhibits temperature scaling and, from fits to the theoretical scaling form, extract values for the effective charge and the interaction parameter of the quasi-particles. Ranges of values obtained are consistent with those predicted by certain models of the 5/2 state. PMID- 18420898 TI - Ocean science. Marine calcifiers in a high-CO2 ocean. PMID- 18420899 TI - Reconstitution of contractile FtsZ rings in liposomes. AB - FtsZ is a tubulin homolog and the major cytoskeletal protein in bacterial cell division. It assembles into the Z ring, which contains FtsZ and a dozen other division proteins, and constricts to divide the cell. We have constructed a membrane-targeted FtsZ (FtsZ-mts) by splicing an amphipathic helix to its C terminus. When mixed with lipid vesicles, FtsZ-mts was incorporated into the interior of some tubular vesicles. There it formed multiple Z rings that could move laterally in both directions along the length of the liposome and coalesce into brighter Z rings. Brighter Z rings produced visible constrictions in the liposome, suggesting that FtsZ itself can assemble the Z ring and generate a force. No other proteins were needed for assembly and force generation. PMID- 18420900 TI - Fracture propagation to the base of the Greenland Ice Sheet during supraglacial lake drainage. AB - Surface meltwater that reaches the base of an ice sheet creates a mechanism for the rapid response of ice flow to climate change. The process whereby such a pathway is created through thick, cold ice has not, however, been previously observed. We describe the rapid (<2 hours) drainage of a large supraglacial lake down 980 meters through to the bed of the Greenland Ice Sheet initiated by water driven fracture propagation evolving into moulin flow. Drainage coincided with increased seismicity, transient acceleration, ice-sheet uplift, and horizontal displacement. Subsidence and deceleration occurred over the subsequent 24 hours. The short-lived dynamic response suggests that an efficient drainage system dispersed the meltwater subglacially. The integrated effect of multiple lake drainages could explain the observed net regional summer ice speedup. PMID- 18420901 TI - Seasonal speedup along the western flank of the Greenland Ice Sheet. AB - It has been widely hypothesized that a warmer climate in Greenland would increase the volume of lubricating surface meltwater reaching the ice-bedrock interface, accelerating ice flow and increasing mass loss. We have assembled a data set that provides a synoptic-scale view, spanning ice-sheet to outlet-glacier flow, with which to evaluate this hypothesis. On the ice sheet, these data reveal summer speedups (50 to 100%) consistent with, but somewhat larger than, earlier observations. The relative speedup of outlet glaciers, however, is far smaller (<15%). Furthermore, the dominant seasonal influence on Jakobshavn Isbrae's flow is the calving front's annual advance and retreat. With other effects producing outlet-glacier speedups an order of magnitude larger, seasonal melt's influence on ice flow is likely confined to those regions dominated by ice-sheet flow. PMID- 18420903 TI - Climate change. IPCC tunes up for its next report aiming for better, timely results. PMID- 18420902 TI - New career paths for scientists. PMID- 18420904 TI - Glaciology. Greenland ice slipping away but not all that quickly. PMID- 18420905 TI - Archaeology. Team unveils Mideast archaeology peace plan. PMID- 18420906 TI - International aid. As food prices rise, U.S. support for agricultural centers wilts. PMID- 18420907 TI - Scientific publishing. Croatian editors fight with medical school over journal's fate. PMID- 18420908 TI - Regulatory science. Changes to EPA toxicology--speed or delay? PMID- 18420909 TI - Energy. The greening of synfuels. PMID- 18420910 TI - Oceanography. Watery echoes give clues to the past and future of the seas. PMID- 18420911 TI - Virology. Mapmaker for the world of influenza. PMID- 18420913 TI - Condensed-matter physics. The mad dash to make light crystals. PMID- 18420912 TI - Virology. Coming out of Asia--year in, year out. PMID- 18420914 TI - The Gonzo Scientist. Scientists, we need your swords! PMID- 18420915 TI - Coral adaptation in the face of climate change. PMID- 18420916 TI - Freshwater forcing: will history repeat itself? PMID- 18420918 TI - Mathematics. Frustration in complexity. PMID- 18420917 TI - Ecology. Agriculture at a crossroads. PMID- 18420919 TI - Geoscience. Natural complexity. PMID- 18420920 TI - Applied physics. Graphene nanoelectronics. PMID- 18420921 TI - Medicine. The ultimate model organism. PMID- 18420922 TI - Cell signaling. Wnt moves beyond the canon. PMID- 18420923 TI - Music theory. Geometrical music theory. PMID- 18420924 TI - Qinghaosu (artemisinin): the price of success. AB - Artemisinin and its derivatives have become essential components of antimalarial treatment. These plant-derived peroxides are unique among antimalarial drugs in killing the young intraerythrocytic malaria parasites, thereby preventing their development to more pathological mature stages. This results in rapid clinical and parasitological responses to treatment and life-saving benefit in severe malaria. Artemisinin combination treatments (ACTs) are now first-line drugs for uncomplicated falciparum malaria, but access to ACTs is still limited in most malaria-endemic countries. Improved agricultural practices, selection of high yielding hybrids, microbial production, and the development of synthetic peroxides will lower prices. A global subsidy would make these drugs more affordable and available. ACTs are central to current malaria elimination initiatives, but there are concerns that tolerance to artemisinins may be emerging in Cambodia. PMID- 18420925 TI - The movement of aquatic mercury through terrestrial food webs. AB - Mercury has contaminated rivers worldwide, with health consequences for aquatic organisms and humans who consume them. Researchers have focused on aquatic birds as sentinels for mercury. However, trophic transfer between adjacent ecosystems could lead to the export of aquatic mercury to terrestrial habitats. Along a mercury-contaminated river in Virginia, United States, terrestrial birds had significantly elevated levels of mercury in their blood, similar to their aquatic feeding counterparts. Diet analysis revealed that spiders delivered much of the dietary mercury. We conclude that aquatic mercury pollution can move into terrestrial habitats, where it biomagnifies to levels in songbirds that may cause adverse effects. Rivers contaminated with mercury may pose a threat to the many bird species that feed on predatory invertebrates in adjacent riparian habitats. PMID- 18420926 TI - Phytoplankton calcification in a high-CO2 world. AB - Ocean acidification in response to rising atmospheric CO2 partial pressures is widely expected to reduce calcification by marine organisms. From the mid Mesozoic, coccolithophores have been major calcium carbonate producers in the world's oceans, today accounting for about a third of the total marine CaCO3 production. Here, we present laboratory evidence that calcification and net primary production in the coccolithophore species Emiliania huxleyi are significantly increased by high CO2 partial pressures. Field evidence from the deep ocean is consistent with these laboratory conclusions, indicating that over the past 220 years there has been a 40% increase in average coccolith mass. Our findings show that coccolithophores are already responding and will probably continue to respond to rising atmospheric CO2 partial pressures, which has important implications for biogeochemical modeling of future oceans and climate. PMID- 18420927 TI - The global circulation of seasonal influenza A (H3N2) viruses. AB - Antigenic and genetic analysis of the hemagglutinin of approximately 13,000 human influenza A (H3N2) viruses from six continents during 2002-2007 revealed that there was continuous circulation in east and Southeast Asia (E-SE Asia) via a region-wide network of temporally overlapping epidemics and that epidemics in the temperate regions were seeded from this network each year. Seed strains generally first reached Oceania, North America, and Europe, and later South America. This evidence suggests that once A (H3N2) viruses leave E-SE Asia, they are unlikely to contribute to long-term viral evolution. If the trends observed during this period are an accurate representation of overall patterns of spread, then the antigenic characteristics of A (H3N2) viruses outside E-SE Asia may be forecast each year based on surveillance within E-SE Asia, with consequent improvements to vaccine strain selection. PMID- 18420928 TI - Generalized voice-leading spaces. AB - Western musicians traditionally classify pitch sequences by disregarding the effects of five musical transformations: octave shift, permutation, transposition, inversion, and cardinality change. We model this process mathematically, showing that it produces 32 equivalence relations on chords, 243 equivalence relations on chord sequences, and 32 families of geometrical quotient spaces, in which both chords and chord sequences are represented. This model reveals connections between music-theoretical concepts, yields new analytical tools, unifies existing geometrical representations, and suggests a way to understand similarity between chord types. PMID- 18420929 TI - Picosecond coherent optical manipulation of a single electron spin in a quantum dot. AB - Most schemes for quantum information processing require fast single-qubit operations. For spin-based qubits, this involves performing arbitrary coherent rotations of the spin state on time scales much faster than the spin coherence time. By applying off-resonant, picosecond-scale optical pulses, we demonstrated the coherent rotation of a single electron spin through arbitrary angles up to pi radians. We directly observed this spin manipulation using time-resolved Kerr rotation spectroscopy and found that the results are well described by a model that includes the electronnuclear spin interaction. Measurements of the spin rotation as a function of laser detuning and intensity confirmed that the optical Stark effect is the operative mechanism. PMID- 18420930 TI - Chaotic Dirac billiard in graphene quantum dots. AB - The exceptional electronic properties of graphene, with its charge carriers mimicking relativistic quantum particles and its formidable potential in various applications, have ensured a rapid growth of interest in this new material. We report on electron transport in quantum dot devices carved entirely from graphene. At large sizes (>100 nanometers), they behave as conventional single electron transistors, exhibiting periodic Coulomb blockade peaks. For quantum dots smaller than 100 nanometers, the peaks become strongly nonperiodic, indicating a major contribution of quantum confinement. Random peak spacing and its statistics are well described by the theory of chaotic neutrino billiards. Short constrictions of only a few nanometers in width remain conductive and reveal a confinement gap of up to 0.5 electron volt, demonstrating the possibility of molecular-scale electronics based on graphene. PMID- 18420931 TI - Atomlike, hollow-core-bound molecular orbitals of C60. AB - The atomic electron orbitals that underlie molecular bonding originate from the central Coulomb potential of the atomic core. We used scanning tunneling microscopy and density functional theory to explore the relation between the nearly spherical shape and unoccupied electronic structure of buckminsterfullerene (C60) molecules adsorbed on copper surfaces. Besides the known pi* antibonding molecular orbitals of the carbon-atom framework, above 3.5 electron volts we found atomlike orbitals bound to the core of the hollow C60 cage. These "superatom" states hybridize like the s and p orbitals of hydrogen and alkali atoms into diatomic molecule-like dimers and free-electron bands of one-dimensional wires and two-dimensional quantum wells in C60 aggregates. We attribute the superatom states to the central potential binding an electron to its screening charge, a property expected for hollow-shell molecules derived from layered materials. PMID- 18420932 TI - The chemical genomic portrait of yeast: uncovering a phenotype for all genes. AB - Genetics aims to understand the relation between genotype and phenotype. However, because complete deletion of most yeast genes ( approximately 80%) has no obvious phenotypic consequence in rich medium, it is difficult to study their functions. To uncover phenotypes for this nonessential fraction of the genome, we performed 1144 chemical genomic assays on the yeast whole-genome heterozygous and homozygous deletion collections and quantified the growth fitness of each deletion strain in the presence of chemical or environmental stress conditions. We found that 97% of gene deletions exhibited a measurable growth phenotype, suggesting that nearly all genes are essential for optimal growth in at least one condition. PMID- 18420933 TI - Wnt5a control of cell polarity and directional movement by polarized redistribution of adhesion receptors. AB - Mechanisms by which Wnt pathways integrate the organization of receptors, organelles, and cytoskeletal proteins to confer cell polarity and directional cell movement are incompletely understood. We show that acute responses to Wnt5a involve recruitment of actin, myosin IIB, Frizzled 3, and melanoma cell adhesion molecule into an intracellular structure in a melanoma cell line. In the presence of a chemokine gradient, this Wnt-mediated receptor-actin-myosin polarity (W RAMP) structure accumulates asymmetrically at the cell periphery, where it triggers membrane contractility and nuclear movement in the direction of membrane retraction. The process requires endosome trafficking, is associated with multivesicular bodies, and is regulated by Wnt5a through the small guanosine triphosphatases Rab4 and RhoB. Thus, cell-autonomous mechanisms allow Wnt5a to control cell orientation, polarity, and directional movement in response to positional cues from chemokine gradients. PMID- 18420934 TI - Recapitulation of IVIG anti-inflammatory activity with a recombinant IgG Fc. AB - It is well established that high doses of monomeric immunoglobulin G (IgG) purified from pooled human plasma [intravenous immunoglobulin (IVIG)] confer anti inflammatory activity in a variety of autoimmune settings. However, exactly how those effects are mediated is not clear because of the heterogeneity of IVIG. Recent studies have demonstrated that the anti-inflammatory activity of IgG is completely dependent on sialylation of the N-linked glycan of the IgG Fc fragment. Here we determine the precise glycan requirements for this anti inflammatory activity, allowing us to engineer an appropriate IgG1 Fc fragment, and thus generate a fully recombinant, sialylated IgG1 Fc with greatly enhanced potency. This therapeutic molecule precisely defines the biologically active component of IVIG and helps guide development of an IVIG replacement with improved activity and availability. PMID- 18420935 TI - Structural basis of toll-like receptor 3 signaling with double-stranded RNA. AB - Toll-like receptor 3 (TLR3) recognizes double-stranded RNA (dsRNA), a molecular signature of most viruses, and triggers inflammatory responses that prevent viral spread. TLR3 ectodomains (ECDs) dimerize on oligonucleotides of at least 40 to 50 base pairs in length, the minimal length required for signal transduction. To establish the molecular basis for ligand binding and signaling, we determined the crystal structure of a complex between two mouse TLR3-ECDs and dsRNA at 3.4 angstrom resolution. Each TLR3-ECD binds dsRNA at two sites located at opposite ends of the TLR3 horseshoe, and an intermolecular contact between the two TLR3 ECD C-terminal domains coordinates and stabilizes the dimer. This juxtaposition could mediate downstream signaling by dimerizing the cytoplasmic Toll interleukin 1 receptor (TIR) domains. The overall shape of the TLR3-ECD does not change upon binding to dsRNA. PMID- 18420936 TI - Divergence of quaternary structures among bacterial flagellar filaments. AB - It has been widely assumed that the atomic structure of the flagellar filament from Salmonella typhimurium serves as a model for all bacterial flagellar filaments given the sequence conservation in the coiled-coil regions responsible for polymerization. On the basis of electron microscopic images, we show that the flagellar filaments from Campylobacter jejuni have seven protofilaments rather than the 11 in S. typhimurium. The vertebrate Toll-like receptor 5 (TLR5) recognizes a region of bacterial flagellin that is involved in subunit-subunit assembly in Salmonella and many other pathogenic bacteria, and this short region has diverged in Campylobacter and related bacteria, such as Helicobacter pylori, which are not recognized by TLR5. The driving force in the change of quaternary structure between Salmonella and Campylobacter may have been the evasion of TLR5. PMID- 18420937 TI - The antidepressant fluoxetine restores plasticity in the adult visual cortex. AB - We investigated whether fluoxetine, a widely prescribed medication for treatment of depression, restores neuronal plasticity in the adult visual system of the rat. We found that chronic administration of fluoxetine reinstates ocular dominance plasticity in adulthood and promotes the recovery of visual functions in adult amblyopic animals, as tested electrophysiologically and behaviorally. These effects were accompanied by reduced intracortical inhibition and increased expression of brain-derived neurotrophic factor in the visual cortex. Cortical administration of diazepam prevented the effects induced by fluoxetine, indicating that the reduction of intracortical inhibition promotes visual cortical plasticity in the adult. Our results suggest a potential clinical application for fluoxetine in amblyopia as well as new mechanisms for the therapeutic effects of antidepressants and for the pathophysiology of mood disorders. PMID- 18420938 TI - Local and systemic glucocorticoid metabolism in inflammatory arthritis. AB - BACKGROUND: Isolated, primary synovial fibroblasts generate active glucocorticoids through expression of 11beta-hydroxysteroid dehydrogenase type 1 (11beta-HSD1). This enzyme produces cortisol from inactive cortisone (and prednisolone from prednisone). OBJECTIVE: To determine how intact synovial tissue metabolises glucocorticoids and to identify the local and systemic consequences of this activity by examination of glucocorticoid metabolism in patients with rheumatoid arthritis (RA). METHODS: Synovial tissue was taken from patients with RA during joint replacement surgery. Glucocorticoid metabolism in explants was assessed by thin-layer chromatography and specific enzyme inhibitors. RT-PCR and immunohistochemistry were used to determine expression and distribution of 11beta HSD enzymes. Systemic glucocorticoid metabolism was examined in patients with RA using gas chromatography/mass spectrometry. RESULTS: Synovial tissue synthesised cortisol from cortisone, confirming functional 11beta-HSD1 expression. In patients with RA, enzyme activity correlated with donor erythrocyte sedimentation rate (ESR). Synovial tissues could also convert cortisol back to cortisone. Inhibitor studies and immunohistochemistry suggested this was owing to 11beta HSD2 expression in synovial macrophages, whereas 11beta-HSD1 expression occurred primarily in fibroblasts. Synovial fluids exhibited lower cortisone levels than matched serum samples, indicating net local steroid activation. Urinary analyses indicated high 11beta-HSD1 activity in untreated patients with RA compared with controls and a significant correlation between total body 11beta-HSD1 activity and ESR. CONCLUSIONS: Synovial tissue metabolises glucocorticoids, the predominant effect being glucocorticoid activation, and this increases with inflammation. Endogenous glucocorticoid production in the joint is likely to have an impact on local inflammation and bone integrity. PMID- 18420939 TI - Remission achieved after 2 years treatment with low-dose prednisolone in addition to disease-modifying anti-rheumatic drugs in early rheumatoid arthritis is associated with reduced joint destruction still present after 4 years: an open 2 year continuation study. AB - OBJECTIVE: To evaluate if remission induced by low-dose prednisolone during the first 2 years of rheumatoid arthritis (RA) in the BARFOT glucocorticoid (GC) study had a sustained effect on radiological damage for a total of 4 years. METHODS: A total of 150 of 211 eligible patients with RA who had been randomised to the 7.5 mg prednisolone group (P) or no prednisolone group (NoP) in addition to the initial disease-modifying antirheumatic drugs were included. Radiographs of hands and feet were scored using the Sharp-van der Heijde scoring method. A patient was considered to be in remission if the 28-joint count disease activity score was <2.6. RESULTS: Mean (SD) age was 53 (14) and 57 (12) years for the patients in the P and NoP groups, respectively. 64% were female, 64% rheumatoid factor positive, and disease duration at baseline was 6 months. At 2 years the proportion of patients in remission in the P and NoP groups was 55 vs 30%, p = 0.003. Longitudinal analysis showed that over the entire course of the disease, patients on prednisolone had a higher probability of being in remission. Patients in remission at 2 years, compared with those not in remission, had significantly lower total Sharp score, erosion score and joint space narrowing score at 2 and 4 years. The changes in bone mineral density during the 4 years did not differ between those in remission and those with active disease, and were similar in the two treatment groups. CONCLUSIONS: Prednisolone 7.5 mg daily in addition to disease-modifying anti-rheumatic drugs increases the rate of remission in patients with early RA, which has a beneficial and sustained effect on radiological damage. PMID- 18420940 TI - Management of Behcet disease: a systematic literature review for the European League Against Rheumatism evidence-based recommendations for the management of Behcet disease. AB - OBJECTIVES: To present and analyse the literature sources regarding the management of Behcet disease (BD) identified during the systematic literature research, which formed the basis for the European League Against Rheumatism (EULAR) evidence-based recommendations for the management of BD. METHODS: Problem areas and related keywords regarding the management of BD were determined by the multidisciplinary expert committee commissioned by EULAR for developing the recommendations. A systematic literature research was performed using MedLine and Cochrane Library resources through to December 2006. Meta-analyses, systematic reviews, randomised controlled trials (RCTs), open studies, observational studies, case control studies and case series' involving > or = 5 patients were included. For each intervention the effect size and number needed to treat were calculated for efficacy. Odds ratios and numbers needed to harm were calculated for safety issues of different treatment modalities where possible. RESULTS: The literature research yielded 137 articles that met the inclusion criteria; 20 of these were RCTs. There was good evidence supporting the use of azathioprine and cyclosporin A in eye involvement and interferon (IFN)alpha in mucocutaneous involvement. There were no RCTs with IFNalpha or tumour necrosis factor (TNF)alpha antagonists in eye involvement. Similarly controlled data for the management of vascular, gastrointestinal and neurological involvement is lacking. CONCLUSION: Properly designed, controlled studies (new and confirmatory) are still needed to guide us in managing BD. PMID- 18420941 TI - Interferon-beta signaling is enhanced in patients with insufficient coronary collateral artery development and inhibits arteriogenesis in mice. AB - Stimulation of collateral artery growth in patients has been hitherto unsuccessful, despite promising experimental approaches. Circulating monocytes are involved in the growth of collateral arteries, a process also referred to as arteriogenesis. Patients show a large heterogeneity in their natural arteriogenic response on arterial obstruction. We hypothesized that circulating cell transcriptomes would provide mechanistic insights and new therapeutic strategies to stimulate arteriogenesis. Collateral flow index was measured in 45 patients with single-vessel coronary artery disease, separating collateral responders (collateral flow index, >0.21) and nonresponders (collateral flow index, < or 1). Isolated monocytes were stimulated with lipopolysaccharide or taken into macrophage culture for 20 hours to mimic their phenotype during arteriogenesis. Genome-wide mRNA expression analysis revealed 244 differentially expressed genes (adjusted P, <0.05) in stimulated monocytes. Interferon (IFN)-beta and several IFN-related genes showed increased mRNA levels in 3 of 4 cellular phenotypes from nonresponders. Macrophage gene expression correlated with stimulated monocytes, whereas resting monocytes and progenitor cells did not display differential gene regulation. In vitro, IFN-beta dose-dependently inhibited smooth muscle cell proliferation. In a murine hindlimb model, perfusion measured 7 days after femoral artery ligation showed attenuated arteriogenesis in IFN-beta-treated mice compared with controls (treatment versus control: 31.5+/-1.2% versus 41.9+/-1.9% perfusion restoration, P<0.01). In conclusion, patients with differing arteriogenic response as measured with collateral flow index display differential transcriptomes of stimulated monocytes. Nonresponders show increased expression of IFN-beta and its downstream targets, and IFN-beta attenuates proliferation of smooth muscle cells in vitro and hampers arteriogenesis in mice. Inhibition of IFN-beta signaling may serve as a novel approach for the stimulation of collateral artery growth. PMID- 18420942 TI - Intramural foci during long duration fibrillation in the pig ventricle. AB - For more than 50 years, it has been assumed that ventricular fibrillation (VF) is maintained solely by reentry in the working myocardium. This hypothesis has never been tested by recording VF with electrodes spaced sufficiently close to map activation sequences in 3D. We recorded the first 10 minutes of electrically induced VF from the anterior left ventricular (LV) free wall near the insertion of the anterior papillary muscle in 6 pigs. A 3D transmural unipolar electrode array consisting of a 9x9 array of needles with 2-mm spacing and 6 electrodes 2 mm apart on each needle was used for recordings. Automatic analyses were performed to recognize 3D reentry and foci. Our results showed that intramural reentry is present early but not late during VF in the mapped region. The incidence of reentry in working myocardium decreases almost to 0 after 3 minutes of VF. In contrast, intramural foci are present during early VF and, as VF continues, increase in incidence, so that by 10 minutes of VF, 27% of wavefronts arise from intramural foci. These results suggest that, particularly after the first 3 minutes of VF, mechanisms other than local reentry in the working myocardium maintain VF in the anterior LV free wall near the root of the anterior papillary muscle. Intramural foci may play an important role in later VF maintenance. It remains to be determined if these foci arise from Purkinje fibers attributable to abnormal automaticity, afterdepolarizations, or reentry. PMID- 18420943 TI - ADAM10 regulates endothelial permeability and T-Cell transmigration by proteolysis of vascular endothelial cadherin. AB - Vascular endothelial (VE)-cadherin is the major adhesion molecule of endothelial adherens junctions. It plays an essential role in controlling endothelial permeability, vascular integrity, leukocyte transmigration, and angiogenesis. Elevated levels of soluble VE-cadherin are associated with diseases like coronary atherosclerosis. Previous data showed that the extracellular domain of VE cadherin is released by an unknown metalloprotease activity during apoptosis. In this study, we used gain-of-function analyses, inhibitor studies, and RNA interference experiments to analyze the proteolytic release of VE-cadherin in human umbilical vein endothelial cells (HUVECs). We found that VE-cadherin is specifically cleaved by the disintegrin and metalloprotease ADAM10 in its ectodomain, releasing a soluble fragment and generating a carboxyl-terminal membrane-bound stub, which is a substrate for a subsequent gamma-secretase cleavage. This ADAM10-mediated proteolysis could be induced by Ca(2+) influx and staurosporine treatment, indicating that ADAM10-mediated VE-cadherin cleavage contributes to the dissolution of adherens junctions during endothelial cell activation and apoptosis, respectively. In contrast, protein kinase C activation or inhibition did not modulate VE-cadherin processing. Increased ADAM10 expression was functionally associated with an increase in endothelial permeability. Remarkably, our data indicate that ADAM10 activity also contributes to the thrombin-induced decrease of endothelial cell-cell adhesion. Moreover, knockdown of ADAM10 in HUVECs as well as in T cells by small interfering RNA impaired T-cell transmigration. Taken together, our data identify ADAM10 as a novel regulator of vascular permeability and demonstrate a hitherto unknown function of ADAM10 in the regulation of VE-cadherin-dependent endothelial cell functions and leukocyte transendothelial migration. PMID- 18420944 TI - Chromogranin B regulates calcium signaling, nuclear factor kappaB activity, and brain natriuretic peptide production in cardiomyocytes. AB - Altered regulation of signaling pathways can lead to pathologies including cardiac hypertrophy and heart failure. We report that neonatal and adult cardiomyocytes express chromogranin B (CGB), a Ca(2+) binding protein that modulates Ca(2+) release by the inositol 1,4,5-trisphosphate receptor (InsP(3)R). Using fluorescent Ca(2+) indicator dyes, we found that CGB regulates InsP(3) dependent Ca(2+) release in response to angiotensin II, an octapeptide hormone that promotes cardiac hypertrophy. ELISA experiments and luciferase reporter assays identified angiotensin II as a potent inducer of brain natriuretic peptide (BNP), a hormone that recently emerged as an important biomarker in cardiovascular disease. CGB was found to regulate angiotensin II-stimulated and basal secretion, expression and promoter activity of BNP that depend on the InsP(3)R. Moreover, we provide evidence that CGB acts via the transcription activity of nuclear factor kappaB in an InsP(3)/Ca(2+)-dependent manner but independent of nuclear factor of activated T cells. In vivo experiments further showed that cardiac hypertrophy induced by angiotensin II, a condition characterized by increased ventricular BNP production, is associated with upregulation of ventricular CGB expression. Overexpression of CGB in cardiomyocytes, in turn, induced the BNP promoter. The evidence presented in this study identifies CGB as a novel regulator of cardiomyocyte InsP(3)/Ca(2+) dependent signaling, nuclear factor kappaB activity, and BNP production. PMID- 18420945 TI - Ste20-related kinase SLK phosphorylates Ser188 of RhoA to induce vasodilation in response to angiotensin II Type 2 receptor activation. AB - The small G protein Rho signaling pathways are recognized as major regulators of cardiovascular functions, and activation of Rho proteins appears to be a common component for the pathogenesis of hypertension and vascular proliferative disorders. Recent evidence suggests that modulation of Rho protein signaling by phosphorylation of Rho proteins provides an additional simple mechanism for coordinating Rho protein functions. Phosphorylation of RhoA by cAMP- or cGMP activated kinase on Ser188 induces cytosolic sequestration of RhoA through increased interaction with guanine dissociation inhibitor, thereby resulting in inhibition of RhoA-dependent functions. Here we show that stimulation of angiotensin II (Ang II) type 2 receptor (AT(2)R) in vascular smooth muscle cells induces Ser188 phosphorylation of RhoA independently of cAMP- or cGMP-activated kinase. We identify the Ser/Thr kinase Ste20-related kinase SLK as a new kinase phosphorylating RhoA on Ser188. Activation of the signaling cascade involving Src homology 2 domain-containing protein-tyrosine phosphatase 1, casein kinase II and SLK is responsible for RhoA phosphorylation and inhibition of RhoA-mediated arterial contraction induced by AT(2)R activation. These results thus identify the molecular mechanism linking AT(2)R to RhoA inhibition and vasodilation. PMID- 18420947 TI - A dual role of the NF-kappaB pathway in neonatal hypoxic-ischemic brain damage. AB - BACKGROUND AND PURPOSE: NF-kappaB is a transcription factor that regulates inflammatory and apoptotic pathways. We described previously that intraperitoneal administration of the NF-kappaB inhibitor TAT-NBD at 0 and 3 hours after neonatal hypoxia-ischemia (HI) markedly reduced brain damage. We hypothesize that timing and duration of NF-kappaB inhibition will be a major factor in determining outcome. METHODS: HI was induced in P7 rats by unilateral carotid artery occlusion and hypoxia. In vivo TAT-NBD effects were determined on cerebral damage, NF-kappaB activity, cytokine expression, and pro- and antiapoptotic molecules. In vitro effects of TAT-NBD were determined using primary neurons and cell lines. RESULTS: HI induced 2 peaks of cerebral NF-kappaB activity at 3 to 6 and 24 hours after HI. Neuroprotective 0/3-hour TAT-NBD treatment only inhibited early NF-kappaB activity. However, inhibition of both early and late NF-kappaB activity by 0/6/12-hour TAT-NBD or only late NF-kappaB activity by 18/21-hour TAT NBD aggravated damage. 0/6/12-hour TAT-NBD did not prevent HI-induced upregulation of cytokines at 24 hours after HI. Protective 0/3-hour TAT-NBD treatment prevented nuclear accumulation of p53 at 24 hours after HI. Nuclear p53 was not reduced after 0/6/12-hour TAT-NBD. Prolonged TAT-NBD increased the proapoptotic factor PUMA and reduced the antiapoptotic factors Bcl-2 and Bcl-xL. Also in neuronal cultures prolonged TAT-NBD exposure overruled protective short term TAT-NBD treatment. CONCLUSIONS: Early NF-kappaB activation contributes to neonatal HI brain damage. Late NF-kappaB provides endogenous neuroprotection and upregulates antiapoptotic molecules. Inhibition of early NF-kappaB activity is neuroprotective only when late NF-kappaB activity is maintained. Moreover, cerebral cytokine production can occur independently of NF-kappaB. PMID- 18420946 TI - A cyclin D2-Rb pathway regulates cardiac myocyte size and RNA polymerase III after biomechanical stress in adult myocardium. AB - Normally, cell cycle progression is tightly coupled to the accumulation of cell mass; however, the mechanisms whereby proliferation and cell growth are linked are poorly understood. We have identified cyclin (Cyc)D2, a G(1) cyclin implicated in mediating S phase entry, as a potential regulator of hypertrophic growth in adult post mitotic myocardium. To examine the role of CycD2 and its downstream targets, we subjected CycD2-null mice to mechanical stress. Hypertrophic growth in response to transverse aortic constriction was attenuated in CycD2-null compared with wild-type mice. Blocking the increase in CycD2 in response to hypertrophic agonists prevented phosphorylation of CycD2-target Rb (retinoblastoma gene product) in vitro, and mice deficient for Rb had potentiated hypertrophic growth. Hypertrophic growth requires new protein synthesis and transcription of tRNA genes by RNA polymerase (pol) III, which increases with hypertrophic signals. This load-induced increase in RNA pol III activity is augmented in Rb-deficient hearts. Rb binds and represses Brf-1 and TATA box binding protein (TBP), subunits of RNA pol III-specific transcription factor B, in adult myocardium under basal conditions. However, this association is disrupted in response to transverse aortic constriction. RNA pol III activity is unchanged in CycD2(-/-) myocardium after transverse aortic constriction, and there is no dissociation of TBP from Rb. These investigations identify an essential role for the CycD2-Rb pathway as a governor of cardiac myocyte enlargement in response to biomechanical stress and, more fundamentally, as a regulator of the load-induced activation of RNA pol III. PMID- 18420948 TI - Variation in the PDE4D gene and ischemic stroke risk: a systematic review and meta-analysis on 5200 cases and 6600 controls. AB - BACKGROUND AND PURPOSE: PDE4D was identified as the first novel gene associated with ischemic stroke risk. Replication studies have produced conflicting results, but many have been small and underpowered. Meta-analysis provides a method to combine this data and determine in a larger sample size whether the association with PDE4D can be replicated. METHODS: A meta-analysis of all PDE4D variants investigated in relation to ischemic stroke has been undertaken. Analysis of any variant appearing in 2 or more replication studies was included; this comprised 6 single nucleotide polymorphisms together with allele 0 of minisatellite AC008818 and the G0 haplotype. A total of 16 studies were identified, allowing examination of up to 5216 cases and 6615 controls for a single variant. Analyses were performed including all data, excluding data from the original report (providing true replication data), and for individual stroke subtypes and limited to white ethnicity. RESULTS: No individual single nucleotide polymorphism was associated with all ischemic stroke cases. Allele 0 of AC008818 and haplotype G0 carriers was associated with increased risk (relative risk, 1.12; 95 CI, 1.01 to 1.25; P=0.03 and relative risk, 1.18; 95% CI, 1.05 to 1.33; P=0.007), but these associations became nonsignificant after exclusion of the original study from the analysis (relative risk, 1.06; 95% CI, 0.94 to 1.20; P=0.34 and relative risk, 1.16; 95% CI, 1.00 to 1.34; P=0.06, respectively). Analyzing only whites, the majority of cases studied, did not result in a significant association for any analysis. Few robust associations were found with individual stroke subtypes. CONCLUSIONS: No genetic variant examined in PDE4D showed a robust and reproducible association to ischemic stroke. Any association that may exist is likely to be weak and potentially restricted to specific populations. PMID- 18420949 TI - Symptomatic intracranial atherosclerotic disease: what is the best treatment option? PMID- 18420950 TI - Early diffusion-weighted MRI as a predictor of caspase-3 activation after hypoxic ischemic insult in neonatal rodents. AB - BACKGROUND AND PURPOSE: Neonatal encephalopathy in human babies is a serious condition associated with permanent neurological deficits. Diffusion-weighted MRI (DWI) is increasingly used for early diagnosis of brain injury in human babies. The relationship between the presence of DWI abnormalities and cellular injury, including apoptosis, during the neonatal period are not well understood. We asked whether the extent of injury depicted on DWI can predict the presence of caspase 3 activation, a quantitative marker of apoptotic injury, after hypoxia-ischemia (H-I) in postnatal day 7 rats. METHODS: Injury volume was determined by DWI at 2 hours, 24 hours, and 7 days after H-I and compared with histology. Caspase-3 activation and microgliosis were determined at 24 hours post-H-I. RESULTS: DWI defined lesions (eg, decreased apparent diffusion coefficient) at 24 hours post-H I correlated with a major increase in caspase-3 activity in the injured hemisphere and predicted injury. A modest but significant increase in caspase-3 activity occurred in the cortex of rats that had no apparent diffusion coefficient decrease in the injured hemisphere but had unilaterally enlarged regions of high apparent diffusion coefficient at the ipsilateral ventricle/white matter interface. Caspase-3 activity was similar in both hemispheres in pups with unchanged DWI. CONCLUSIONS: Abnormal DWI signal at 24 hours post-H-I is predictive of caspase-3 activation and can be used as an indicator that injury involving an apoptotic-like mechanism is present. Our data also suggest that the presence of an enlarged unilateral region with high apparent diffusion coefficient at the ventricle/white matter interface without significant apparent diffusion coefficient decrease in the cortex is a sign of modest caspase-3 activation after H-I. PMID- 18420951 TI - Local intraarterial fibrinolysis administered in aliquots for the treatment of central retinal artery occlusion: the Johns Hopkins Hospital experience. AB - BACKGROUND AND PURPOSE: Central retinal artery occlusion results in acute visual loss with poor spontaneous recovery. Current standard therapies do not alter the natural history of disease. Several open-label clinical studies using continuous infusion of thrombolytic agents have suggested that local intraarterial fibrinolysis (LIF) is efficacious in the treatment of central retinal artery occlusion. The aim is to compare the visual outcome in patients with acute central retinal artery occlusion of presumed thromboembolic etiology treated with LIF administered in aliquots with that of patients treated with standard therapy. METHODS: We conducted a single-center, nonrandomized interventional study of consecutive patients with acute central retinal artery occlusion from July 1999 to July 2006. RESULTS: Twenty-one patients received LIF and 21 received standard therapy. Seventy-six percent of subjects in the LIF group had a visual acuity improvement of one line or more compared with 33% in the standard therapy group (P=0.012, Fisher exact). Multivariate logistic regression controlling for gender, history of prior stroke/transient ischemic attack, and history of hypercholesterolemia showed that patients who received tissue plasminogen activator were 36 times more likely to have improvement in visual acuity (P=0.0001) after adjusting for these covariates. Post hoc analysis showed that patients who received tissue plasminogen activator were 13 times more likely to have improvement in visual acuity of 3 lines or more (P=0.03) and 4.9 times more likely to have a final visual acuity of 20/200 or better (P=0.04). Two groin hematomas were documented in the LIF group. No ischemic strokes, retinal or intracerebral hemorrhages were documented. CONCLUSIONS: LIF administered in aliquots is associated with an improvement in visual acuity compared with standard therapy and has few side effects. PMID- 18420952 TI - Strong neuroprotection by inhibition of NF-kappaB after neonatal hypoxia-ischemia involves apoptotic mechanisms but is independent of cytokines. AB - BACKGROUND AND PURPOSE: Interactions between excitotoxic, inflammatory, and apoptotic pathways determine outcome in hypoxic-ischemic brain damage. The transcription factor NF-kappaB has been suggested to enhance brain damage via stimulation of cytokine production. There is also evidence that NF-kappaB activity is required for neuronal survival. We used the NF-kappaB inhibitor NBD, coupled to TAT to facilitate cerebral uptake, to determine the neuroprotective capacity of NF-kappaB inhibition in neonatal hypoxia-ischemia (HI) and to identify its contribution to cerebral inflammation and damage. METHODS: Brain damage was induced in neonatal rats by unilateral carotid artery occlusion and hypoxia and analyzed immunohistochemically; NF-kappaB activity was analyzed by EMSA. We analyzed cytokine mRNA levels and activation of apoptotic pathways by Western blotting. In vitro effects of TAT-NBD were determined in a neuronal cell line. RESULTS: Inhibition of cerebral NF-kappaB activity by TAT-NBD had a significant neuroprotective effect; brain damage was reduced by more than 80% with a therapeutic window of at least 6 hours. In contrast to earlier suggestions, the protective effect of TAT-NBD did not involve suppression of early cytokine upregulation after HI. Moreover, NF-kappaB inhibition prevented HI induced upregulation and nuclear as well as mitochondrial accumulation of p53, prevented mitochondrial cytochrome-c release and activation of caspase-3. Finally, TAT-NBD could directly increase neuronal survival because TAT-NBD was sufficient to inhibit death in a neuronal cell line. A nonactive mutant peptide did not have any effect. CONCLUSIONS: Inhibition of NF-kappaB has strong neuroprotective effects that involve downregulation of apoptotic molecules but are independent of inhibition of cytokine production. PMID- 18420954 TI - Comparison of 12 risk stratification schemes to predict stroke in patients with nonvalvular atrial fibrillation. AB - BACKGROUND AND PURPOSE: More than a dozen schemes for stratifying stroke risk in patients with nonvalvular atrial fibrillation have been published. Differences among these schemes lead to inconsistent stroke risk estimates for many atrial fibrillation patients, resulting in confusion among clinicians and nonuniform use of anticoagulation. METHODS: Twelve published schemes stratifying stroke risk in patients with nonvalvular atrial fibrillation are analyzed, and observed stroke rates in independent test cohorts are compared with predicted risk status. RESULTS: Seven schemes were based directly on event-rate analyses, whereas 5 resulted from expert consensus. Four considered only clinical features, whereas 7 schemes included echocardiographic variables. The number of variables per scheme ranged from 4 to 8 (median, 6). The most frequently included features were previous stroke/TIA (100% of schemes), patient age (83%), hypertension (83%), and diabetes (83%), and 8 additional variables were included in >/=1 schemes. Based on published test cohorts, all 8 tested schemes stratified stroke risk, but the absolute stroke rates varied widely. Observed rates for those categorized as low risk ranged from 0% to 2.3% per year and those categorized as high risk ranged from 2.5% to 7.9% per year. When applied to the same cohorts, the fractions of patients categorized by the different schemes as low risk varied from 9% to 49% and those categorized by the different schemes as high-risk varied from 11% to 77%. CONCLUSIONS: There are substantial, clinically relevant differences among published schemes designed to stratify stroke risk in patients with atrial fibrillation. Additional research to identify an optimum scheme for primary prevention and subsequent standardization of recommendations may lead to more uniform selection of patients for anticoagulant prophylaxsis. PMID- 18420953 TI - Effect of prophylactic transluminal balloon angioplasty on cerebral vasospasm and outcome in patients with Fisher grade III subarachnoid hemorrhage: results of a phase II multicenter, randomized, clinical trial. AB - BACKGROUND AND PURPOSE: Cerebral vasospasm continues to be a major cause of poor outcome in patients with ruptured aneurysms. Prophylactic Transluminal Balloon Angioplasty (pTBA) appeared to prevent delayed ischemic neurological deficit in a pilot study. A phase II multicenter randomized clinical trial was subsequently designed. METHODS: One hundred and seventy patients with Fisher Grade III subarachnoid hemorrhage were enrolled in the study. Of these, 85 patients were randomized to the treatment group and underwent pTBA within 96 hours after subarachnoid hemorrhage. Main end points of the study included the 3-month dichotomized Glasgow Outcome Score (GOS), development of delayed ischemic neurological deficit (DIND), occurrence of Transcranial Doppler (TCD) vasospasm, and length of stay in the ICU and hospital. RESULTS: The incidence of DIND was lower in the pTBA group (P=0.30) and fewer patients required therapeutic angioplasty to treat DIND (P=0.03). Overall pTBA resulted in an absolute risk reduction of 5.9% and a relative risk reduction of 10.4% unfavorable outcome (P=0.54). Good grade patients had absolute and relative risk reductions of respectively 9.5 and 29.4% (P=0.73). Length of stay in ICU and hospital was similar in both groups. Four patients had a procedure-related vessel perforation, of which three patients died. CONCLUSIONS: While the trial is unsuccessful as defined by the primary end point (GOS), proof of concept is confirmed by these results. Fewer patients tend to develop vasospasm after treatment with pTBA and there is a statistically significantly decreased need for therapeutic angioplasty. pTBA does not improve the poor outcome of patients with Fisher grade III subarachnoid hemorrhage. PMID- 18420955 TI - Stroke in rural areas and small communities. AB - The management of stroke in rural and regional areas is variable in both the developed and developing world. Informed by best-practice guidelines and recommendations for systems of stroke care, adaptable models of care that are appropriate for local needs should be devised for rural and regional settings. This review addresses the issue of the provision of appropriate services in rural and regional settings, with particular attention to the barriers involved, according to the classification of Low Human Development Country (LHDC), Medium Human Development Country (MHDC) and High Human Development Country (HHDC). We discuss the need and feasibility of developing implementing stroke care in rural settings according to best-practice recommendations, within models of care adapted to local conditions. PMID- 18420956 TI - Durability of endovascular therapy for symptomatic intracranial atherosclerosis. AB - BACKGROUND AND PURPOSE: Intracranial angioplasty and stenting are therapeutic options for patients with symptomatic intracranial arterial stenoses intractable to medical therapy. However, the long-term safety and clinical efficacy of these techniques are unknown. We sought to assess the long-term outcome and efficacy of these techniques. METHODS: Procedural data and 30-day outcomes were collected from patients treated with coronary balloons and stents for >/=70% atherostenoses. Clinical and radiographic follow-up data were obtained at 30 days, 6 months, 12 months, and yearly thereafter. RESULTS: Fifty-three patients (median age, 67 years; interquartile range [IQR], 58.75 to 75 years) with 69 arterial lesions were treated during a 7-year period. The technical success rate was 98.6% (68/69), with a reduction of the median percent stenosis from 85% (IQR, 70% to 95%) to 0% (IQR, 0% to 26%). In 76.8% (53/69) of the procedures, a stent was implanted. The 30-day death/stroke rate was 10.1% (7/69) with 1 death, and within a median follow-up of 24 months (IQR, 10.25 to 36.5 months), the transient ischemic attack or stroke rate reached 5.8% (4/69). Restenosis rate at 1 year was 15.9% (11/69) and was symptomatic in 18.2% (2/11). The restenosis rate was 50% for angioplasty (8/16) and 7.5% (4/53) for stenting (hazard ratio=5.02; 95% CI, 1.22 to 20.68). Factors associated with restenosis were vessel size <2.5 mm (hazard ratio=4.78; 95% CI, 1.35 to 16.93) and interventions performed in the setting of an acute stroke (hazard ratio=6.36; 95% CI, 1.78 to 22.56). CONCLUSIONS: Intracranial stenting may reduce the rate of recurrent ischemia in patients in whom medical therapy is unsuccessful and is probably more durable than angioplasty alone. PMID- 18420957 TI - When to expect negative diffusion-weighted images in stroke and transient ischemic attack. AB - BACKGROUND AND PURPOSE: The frequency of DWI negative cerebral ischemia and clinical factors associated with such a circumstance is not well understood. METHODS: We performed MRI including diffusion-weighted imaging (DWI) in patients with stroke and transient ischemic attack (TIA) within 24 hours of symptom onset and again at 30 days. RESULTS: Of 401 patients, 103 (25.6%) had an initial negative DWI study. In the DWI negative group, among the stroke patients, 6/26 (23.1%) had infarcts on follow-up MRI (4 lacunar and 2 posterior circulation syndromes) and 1 had a rMTT deficit. Among the TIA patients, 4/63 (6.3%) showed rMTT deficits and 2/63 (3.2%) had infarcts on follow-up MRI. CONCLUSIONS: Baseline perfusion weighted imaging sequences may detect ischemia in a small proportion of DWI negative cases. Only those with brain stem location or lacunar syndrome were DWI negative initially and yet had a follow-up imaging confirmation of infarct or a final clinical diagnosis of stroke. PMID- 18420958 TI - Unilateral ablation of pre-Botzinger complex disrupts breathing during sleep but not wakefulness. AB - RATIONALE: In adult rats, bilateral ablation of pre-Botzinger complex (preBotC) neurokinin 1-expressing (NK1R) neurons leads to a progressive and irreversible disruption in breathing pattern, initially during sleep, eventually resulting in an ataxic breathing pattern during wakefulness. OBJECTIVES: Here we determine whether ablation of fewer preBotC NK1R neurons leads to a persistent pattern of disordered breathing during sleep but not during wakefulness. METHODS: Adult male Sprague-Dawley rats (n = 12) were instrumented to record diaphragmatic, abdominal, and neck EMG, and EEG. Fourteen days later, a second surgery was performed to stereotaxically microinject into the preBotC on one side the toxin saporin conjugated to substance P (SP-SAP), which selectively ablates NK1R neurons. MEASUREMENTS AND MAIN RESULTS: Postinjection, rats were monitored within a plethysmograph until they were killed (Days 21-51). At Days 6-9 post-unilateral SP-SAP injection, respiratory pattern during sleep, particularly REM sleep, became increasingly disordered, characterized by an increase in frequency of central sleep apnea and hypopneas (36.8 +/- 7.4 episodes/h of REM vs. 6 +/- 2.0 episodes/h in preinjection controls; P < 0.05), whereas breathing during resting wakefulness remained stable. Unlike bilateral SP-SAP-injected rats, an ataxic breathing pattern did not develop during wakefulness. Rats that were monitored up to 51 days post-SP-SAP injection continued to have sleep-disordered breathing; breathing during wakefulness remained relatively stable. Histologic analysis of the ventrolateral medulla confirmed that NK1R neurons within the preBotC on the injected but not on the contralateral side of the medulla were ablated. CONCLUSIONS: Gradual loss of preBotC NK1R neurons may be an underlying factor of sleep-disordered breathing, in particular of central sleep apnea. PMID- 18420959 TI - Lung function loss, smoking, vitamin C intake, and polymorphisms of the glutamate cysteine ligase genes. AB - RATIONALE: Smoking-induced oxidative stress contributes to chronic obstructive pulmonary disease, a lung disease characterized by low lung function and increasing mortality worldwide. The counterbalance for this effect may be provided by, for example, increased intake of the antioxidant vitamin C or endogenously acting antioxidant enzymes like glutamate-cysteine ligase (GCL), which is responsible for glutathione biosynthesis. OBJECTIVES: To investigate associations of functional polymorphisms in GCL subunits (GCLM and GCLC) with lung function level and its longitudinal course, with vitamin C and smoking habits as potential interactive factors. METHODS: Two independent general population samples (Doetinchem, n = 1,152, and Vlagtwedde-Vlaardingen, n = 1,390) with multiple lung function (FEV(1), VC) measurements were genotyped for three polymorphisms (C[-129]T, C[-588]T, and a trinucleotide GAG repeat [TNR]) in the subunits of GCL. Genetic effects on lung function level and decline were estimated using linear regression and linear mixed effect models adjusted for confounders. Findings were further investigated for interactions with vitamin C intake in the Doetinchem cohort. MEASUREMENTS AND MAIN RESULTS: GCLC polymorphisms were significantly associated with lower lung function levels in interaction with pack-years smoked in both cohorts. TNR variants in GCLC were associated with accelerated FEV(1) decline in both cohorts in interaction with pack-years. All significant effects were specifically present in subjects within the lowest tertile of vitamin C intake. CONCLUSIONS: GCLC is a novel susceptibility gene for low level of lung function in two independent populations. We provide suggestive evidence that this occurs due to an interaction between GCLC polymorphisms, smoking, and low vitamin C intake, which all contribute to the oxidative burden. PMID- 18420960 TI - Azithromycin improves macrophage phagocytic function and expression of mannose receptor in chronic obstructive pulmonary disease. AB - RATIONALE: Defective efferocytosis (phagocytic clearance of apoptotic cells) in the airway may perpetuate inflammation via secondary necrosis in chronic obstructive pulmonary disease (COPD). We have previously reported that low-dose azithromycin improved alveolar macrophage (AM) phagocytic function in vitro. OBJECTIVES: We investigated collectins (mannose-binding lectin [MBL] and surfactant protein [SP]-D) and mannose receptor (MR) in COPD and their possible role in the azithromycin-mediated improvement in phagocytosis. METHODS: In vitro effects of azithromycin on AM expression of MR were investigated. MBL, SP-D, and MR were measured in patients with COPD and control subjects. Azithromycin (250 mg orally daily for 5 d then twice weekly for 12 wk) was administered to 11 patients with COPD. Assessments included AM phagocytic ability and expression of MR, MBL, SP-D, bronchial epithelial cell apoptosis, pulmonary function, C-reactive protein, blood/BAL leukocyte counts, cytokine production, and T-cell markers of activation and phenotype. MEASUREMENTS AND MAIN RESULTS: Azithomycin (500 ng/ml) increased MR expression by 50% in vitro. AM MR expression and levels of MBL and SP-D were significantly reduced in patients with COPD compared with control subjects. In patients with COPD, after azithromycin therapy, we observed significantly improved AM phagocytic ability (pre: 9.9%; post: 15.1%), reduced bronchial epithelial cell apoptosis (pre: 30.0%; post: 19.7%), and increased MR and reduced inflammatory markers in the peripheral blood. These findings implicate the MR in the defective phagocytic function of AMs in COPD and as a target for the azithromycin-mediated improvement in phagocytic ability. CONCLUSIONS: Our findings indicate a novel approach to supplement existing therapies in COPD. PMID- 18420961 TI - Surfactant proteins A and D suppress alveolar macrophage phagocytosis via interaction with SIRP alpha. AB - RATIONALE: Efficient removal of apoptotic cells is essential for the resolution of acute pulmonary inflammation. Alveolar macrophages ingest apoptotic cells less avidly than other professional phagocytes at rest but overcome this defect during acute inflammation. Surfactant protein (SP)-A and SP-D are potent modulators of macrophage function and may suppress clearance of apoptotic cells through activation of the transmembrane receptor signal inhibitory regulatory protein alpha (SIRP alpha). OBJECTIVES: To investigate whether binding of SP-A and SP-D to SIRP alpha on alveolar macrophages suppresses apoptotic cell clearance. METHODS: Phagocytosis of apoptotic cells was assessed using macrophages pretreated with SP-A, SP-D, or the collectin-like molecule C1q. Binding of SP-A and SP-D to SIRP alpha was confirmed in vitro using blocking antibodies and fibroblasts transfected with active and mutant SIRP alpha. The effects of downstream molecules SHP-1 and RhoA on phagocytosis were studied using SHP-1 deficient mice, sodium stibogluconate, and a Rho kinase inhibitor. Lipopolysaccharide was given to chimeric mice to study the effects of SP-A and SP D binding on inflammatory macrophages. MEASUREMENTS AND MAIN RESULTS: Preincubation of macrophages with SP-A or SP-D suppressed apoptotic cell clearance. Surfactant suppression of macrophage phagocytosis was reversed by blocking SIRP alpha and inhibiting downstream molecules SHP-1 and RhoA. Macrophages from inflamed lungs ingested apoptotic cells more efficiently than resting alveolar macrophages. Recruited mononuclear phagocytes with low levels of SP-A and SP-D mediated this effect. CONCLUSIONS: SP-A and SP-D tonically inhibit alveolar macrophage phagocytosis by binding SIRP alpha. During acute pulmonary inflammation, defects in apoptotic cell clearance are overcome by recruited mononuclear phagocytes. PMID- 18420962 TI - Anemia, blood loss, and blood transfusions in North American children in the intensive care unit. AB - RATIONALE: Minimizing exposure of children to blood products is desirable. OBJECTIVES: We aimed to understand anemia development, blood loss, and red blood cell (RBC) transfusions in the pediatric intensive care unit (PICU). METHODS: Prospective, multicenter, 6-month observational study in 30 PICUs. Data were collected on consecutive children (<18 yr old) in the PICU for 48 hours or more. MEASUREMENTS AND MAIN RESULTS: Anemia development, blood loss, and RBC transfusions were measured. A total of 977 children were enrolled. Most (74%) children were anemic in the PICU (33% on admission, 41% developed anemia). Blood draws accounted for 73% of daily blood loss; median loss was 5.0 ml/day. Forty nine percent of children received transfusions; 74% of first transfusions were on Days 1-2. After adjusting for age and illness severity, compared with nontransfused children, children who underwent transfusion had significantly longer days of mechanical ventilation (2.1 d, P < 0.001) and PICU stay (1.8 d, P = 0.03), and had increased mortality (odds ratio [OR], 11.6; 95% confidence interval [CI], 1.43-90.9; P = 0.02), nosocomial infections (OR, 1.9; 95% CI, 1.2 3.0; P = 0.004), and cardiorespiratory dysfunction (OR, 2.1; 95% CI, 1.5-3.0; P < 0.001). High blood loss per kilogram body weight from blood draws (OR, 1.11; 95% CI, 1.03-1.2; P = 0.01) was associated with RBC transfusion more than 48 hours after admission. The most common indication for transfusion was low hemoglobin (42%). Pretransfusion hemoglobin values varied greatly (mean, 9.7 +/- 2.7 g/dl). CONCLUSIONS: Critically ill children are at significant risk for developing anemia and receiving blood transfusions. Transfusion in the PICU was associated with worse outcomes. It is imperative to minimize blood loss from blood draws and to set clear transfusion thresholds. PMID- 18420963 TI - Mapping of a novel susceptibility locus suggests a role for MC3R and CTSZ in human tuberculosis. AB - RATIONALE: Tuberculosis remains a major cause of morbidity and mortality in the developing world. A better understanding of the mechanisms of disease protection could allow novel strategies to disease management and control. OBJECTIVES: To identify human genomic loci with evidence of linkage to tuberculosis susceptibility and, within these loci, to identify individual genes influencing tuberculosis susceptibility. METHODS: Affected sibling pair analysis in South African and Malawian populations. Independent case-control study in West Africa. MEASUREMENTS AND MAIN RESULTS: Two novel putative loci for tuberculosis susceptibility are identified: chromosome 6p21-q23 and chromosome 20q13.31-33- the latter with the strongest evidence for any locus reported to date in human tuberculosis (single point LOD score of 3.1, P = 10(-4), with a maximum likelihood score [MLS] of 2.8). An independent, multistage genetic association study in West African populations mapped this latter region in detail, finding evidence that variation in the melanocortin 3 receptor (MC3R) and cathepsin Z (CTSZ) genes play a role in the pathogenesis of tuberculosis. CONCLUSIONS: These results demonstrate how a genomewide approach to the complex phenotype of human tuberculosis can identify novel targets for further research. PMID- 18420964 TI - Randomized controlled trial of dietary creatine as an adjunct therapy to physical training in chronic obstructive pulmonary disease. AB - RATIONALE: Skeletal muscle strength and bulk are reduced in patients with chronic obstructive pulmonary disease (COPD) and influence quality of life, survival, and utilization of health care resources. Exercise training during pulmonary rehabilitation (PR) can reverse some of these effects. In athletes and healthy elderly individuals, dietary creatine supplementation (CrS) has been shown to augment high-intensity exercise training, thereby increasing muscle mass. OBJECTIVES: This article examines the effect of CrS on functional exercise capacity and muscle performance in people with COPD. METHODS: One hundred subjects with COPD (mean [SD] age, 68.2 [8.2] yr; FEV(1), 44.0 [19.6] %predicted) were randomized to a double-blind, placebo-controlled, parallel group trial of CrS during 7 weeks of PR encompassing aerobic and resistance exercises. Subjects ingested creatine (22 g/d loading for 5 d; maintenance, 3.76 g/d throughout PR) or placebo. Baseline, postloading, and postrehabilitation measurements included pulmonary function, body composition, peripheral muscle strength, and functional performance (shuttle walking tests). A volunteer subgroup (n = 44) had pre- and postloading quadriceps muscle biopsies. MEASUREMENTS AND MAIN RESULTS: Eighty subjects completed the trial (38 creatine, 42 placebo). All outcome measures significantly improved after PR. There were no significant differences between groups post-PR (mean [SD] change in incremental shuttle walk distance, 84 [79] m in the creatine group vs. 83.8 [60] m in the placebo group; P = 1.0; knee extensor work, 19.2 [16] Nm [Newton meters] in the creatine group vs. 19.5 [17] Nm in the placebo group; P = 0.9). Muscle biopsies showed evidence of creatine uptake. CONCLUSIONS: This adequately powered, randomized, placebo-controlled trial shows that CrS does not augment the substantial training effect of multidisciplinary PR for patients with COPD. Clinical trial registered with https://portal.nihr.ac.uk/Pages/NRRArchiveSearch.aspx (NO123138126). PMID- 18420965 TI - Airway responsiveness in mild to moderate childhood asthma: sex influences on the natural history. AB - RATIONALE: Airway responsiveness is a prognostic marker for asthma symptoms in later life. OBJECTIVES: To evaluate characteristics responsible for persistence of airway responsiveness in children with asthma. METHODS: A total of 1,041 children, initially aged 5-12 years, with mild to moderate persistent asthma enrolled in the Childhood Asthma Management Program (CAMP) were studied prospectively for 8.6 +/- 1.8 years with methacholine challenges yearly. MEASUREMENTS AND MAIN RESULTS: Least squares geometric mean models were fit to determine effects of sex and age on airway responsiveness (provocative concentration producing 20% decrease in FEV(1) [PC(20)]). Multiple linear regression analysis was performed to determine factors at baseline and over time, which were associated with PC(20) at end of follow-up. A total of 7,748 methacholine challenges were analyzed. PC(20) increased with age, with boys having greater increase after age 11 years than girls (P < 0.001). The divergence coincided with the mean age for Tanner stage 2. Postpubertal girls had greater airway responsiveness, even after adjustment for FEV(1) and other potential confounders. Although multivariable regression analyses noted a variety of factors that influenced airway responsivness in both sexes, a history of hay fever (beta= -0.30, P = 0.005), respiratory allergy (beta= -0.32, P = 0.006), or recent inhaled corticosteroid usage (beta= -0.18, P = 0.02) were associated with decrements in final log PC(20) only in girls. CONCLUSIONS: Airway responsiveness (PC(20)) is more severe in the postpubertal female with asthma than in males. Although there are factors associated with airway responsiveness in both males and females, sex-specific factors may contribute to new insights into asthma pathogenesis. PMID- 18420966 TI - Platelet-derived growth factor expression and function in idiopathic pulmonary arterial hypertension. AB - RATIONALE: Platelet-derived growth factor (PDGF) promotes the proliferation and migration of pulmonary artery smooth muscle cells (PASMCs), and may play a role in the progression of pulmonary arterial hypertension (PAH), a condition characterized by proliferation of PASMCs resulting in the obstruction of small pulmonary arteries. OBJECTIVES: To analyze the expression and pathogenic role of PDGF in idiopathic PAH. METHODS: PDGF and PDGF receptor mRNA expression was studied by real-time reverse transcription-polymerase chain reaction performed on laser capture microdissected pulmonary arteries from patients undergoing lung transplantation for idiopathic PAH. Immunohistochemistry was used to localize PDGF, PDGF receptors, and phosphorylated PDGFR-beta. The effects of imatinib on PDGF-B-induced proliferation and chemotaxis were tested on human PASMCs. MEASUREMENTS AND MAIN RESULTS: PDGF-A, PDGF-B, PDGFR-alpha, and PDGFR-beta mRNA expression was increased in small pulmonary arteries from patients displaying idiopathic PAH, as compared with control subjects. Western blot analysis revealed a significant increase in protein expression of PDGFR-beta in PAH lungs, as compared with control lungs. In small remodeled pulmonary arteries, PDGF-A and PDGF-B mainly localized to PASMCs and endothelial cells (perivascular inflammatory infiltrates, when present, showed intensive staining), PDGFR-alpha and PDGFR-beta mainly stained PASMCs and to a lesser extent endothelial cells. Proliferating pulmonary vascular lesions stained phosphorylated PDGFR-beta. PDGF BB-induced proliferation and migration of PASMCs were inhibited by imatinib. This effect was not due to PASMC apoptosis. CONCLUSIONS: PDGF may play an important role in human PAH. Novel therapeutic strategies targeting the PDGF pathway should be tested in clinical trials. PMID- 18420968 TI - Dyspnea on exertion in obese women: association with an increased oxygen cost of breathing. AB - RATIONALE: Although exertional dyspnea in obesity is an important and prolific clinical concern, the underlying mechanism remains unclear. OBJECTIVES: To investigate whether dyspnea on exertion in otherwise healthy obese women was associated with an increase in the oxygen cost of breathing or cardiovascular deconditioning. METHODS: Obese women with and without dyspnea on exertion participated in two independent experiments (n = 16 and n = 14). All participants underwent pulmonary function testing, hydrostatic weighing, ratings of perceived breathlessness during cycling at 60 W, and determination of the oxygen cost of breathing during eucapnic voluntary hyperpnea at 40 and 60 L/min. Cardiovascular exercise capacity, fat distribution, and respiratory mechanics were determined in 14 women in experiment 2. Data were analyzed between groups by independent t test, and the relationship between the variables was determined by regression analysis. MEASUREMENTS AND MAIN RESULTS: In both experiments, breathlessness during 60 W cycling was markedly increased in over 37% of the obese women (P < 0.01). Age, height, weight, lung function, and %body fat were not different between the groups in either experiment. In contrast, the oxygen cost of breathing was significantly (P < 0.01) and markedly (38-70%) greater in the obese women with dyspnea on exertion. The oxygen cost of breathing was significantly (P < 0.001) correlated with the rating of perceived breathlessness obtained during the 60 W exercise in experiment 1 (r(2) = 0.57) and experiment 2 (r(2) = 0.72). Peak cardiovascular exercise capacity, fat distribution, and respiratory mechanics were not different between groups in experiment 2. CONCLUSIONS: Dyspnea on exertion is prevalent in otherwise healthy obese women, which seems to be strongly associated with an increased oxygen cost of breathing. Exercise capacity is not reduced in obese women with dyspnea on exertion. PMID- 18420967 TI - Endoglin (CD105) up-regulation in pulmonary microvasculature of ventilated preterm infants. AB - RATIONALE: Preterm infants exposed to mechanical ventilation and oxygen are at risk for bronchopulmonary dysplasia (BPD), a multifactorial chronic lung disorder characterized by arrested alveolar development. Studies have described disruption of microvascular development in BPD, characterized by primitive angioarchitectural patterns reminiscent of the canalicular/saccular stages of lung development. The molecular regulation of this BPD-associated dysangiogenesis remains undetermined. OBJECTIVES: Endoglin (CD105), a hypoxia-inducible transforming growth factor-beta coreceptor, has been implicated as an important regulator of angiogenesis in various neoplastic and nonneoplastic conditions. The aim of this study was to investigate the expression of endoglin and other angiogenesis-related factors in ventilated preterm human lungs. METHODS: We have studied endoglin protein and mRNA expression in postmortem lungs of short-term and long-term ventilated preterm infants. Control subjects were age-matched infants who had lived for less than 1 hour. MEASUREMENTS AND MAIN RESULTS: Lungs of short-term ventilated preterm infants showed significant upregulation of endoglin mRNA and protein levels, immunolocalized to the microvasculature. Similar but more variable endoglin upregulation was noted in lungs of long-term ventilated infants with BPD. The mRNA levels of vascular endothelial growth factor, angiopoietin-1, and their respective receptors were significantly lower in ventilated lungs than in age-matched nonventilated control lungs. CONCLUSIONS: BPD is associated with a shift from traditional angiogenic growth factors (vascular endothelial growth factor, angiopoietin-1) to alternative regulators such as endoglin, which may contribute to BPD-associated microvascular dysangiogenesis. PMID- 18420969 TI - Longitudinal evaluation of airway function 21 years after preterm birth. AB - RATIONALE: There are limited longitudinal data about respiratory morbidity and lung function after preterm birth into adulthood. OBJECTIVES: To determine the evolution of respiratory symptoms, spirometry, and airway hyperresponsiveness of ex-preterm subjects from childhood into adulthood. METHODS: Ex-preterm subjects (median birth weight, 1,440 g; median gestation, 31.5 wk), recruited at birth (not treated with surfactant), had excess respiratory symptoms, airway obstruction, and increased airway hyperresponsiveness in mid-childhood. At a median age of 21.7 years, 60 of these subjects (the index study group) and 50 healthy term control subjects were recruited to determine respiratory morbidity and spirometry. MEASUREMENTS AND MAIN RESULTS: Respiratory symptom questionnaire, spirometry, and methacholine challenge test. The index study group had significantly more respiratory symptoms (16 of 60) than did control subjects (4 of 50) (odds ratio, 4.2; 95% confidence interval, 1.3 to 13.5; P = 0.01), but no significant difference in measured spirometry. Specifically, in the index study group and control subjects, the mean z scores (95% confidence interval of the group difference) for the FEV(1) were -0.60 and -0.58 (-0.44 to 0.49), respectively (P = 0.92); for the forced mid-expiratory flow they were -1.02 and 0.86 (-0.33 to 0.64), respectively (P = 0.52); and for the FVC they were -0.29 and -0.33 (-0.46 to 0.38), respectively (P = 0.85). Ex-preterm adults did not show evidence of increased airway hyperresponsiveness compared with control subjects, 23 and 19%, respectively (P = 0.89). CONCLUSIONS: There are still excess respiratory symptoms 21 years after preterm birth. Reassuringly, this longitudinal study did not show evidence of persistent airway obstruction or airway hyperresponsiveness in ex-preterm adults. PMID- 18420970 TI - Behavior of laying hens on alfalfa crumble molt diets. AB - Several dietary alternatives to feed withdrawal have been proposed to induce a molt in laying hens. This study compared the behavior of laying hens on an alfalfa crumble diet (ALC) to hens that were either on a conventional layer diet (FF) or hens that had feed withdrawn (FW) during a 9-d trial. Each treatment consisted of 24 hens (3 hens per battery cage), and treatment began after a 2 week acclimation period. Video cameras connected to a digital multiplexer recorded the behavior of the hens. The percentages of observations performing nonnutritive pecking, feeder activity, drinking, walking, preening, head movement, and aggression were quantified for two 10-min periods at daily intervals. The FF hens spent significantly more (P < or = 0.05) time drinking than the other treatments, whereas FW hens displayed the most head movements. From d 1 through 7, FW hens walked less than ALC hens except on d 8 when FW hens walked more than ALC and FF hens. On d 4 and 6, the FW hens spent an increased amount of time preening compared with FF hens until the last few days of the molt period. For the most part, FW hens generally displayed more nonnutritive pecking than ALC and FF hens throughout the molt period. However, FW hen visits to the feeders declined as the trial proceeded, whereas ALC and FF hens generally spent more time at the feeder. In summary, the ALC diet showed potential as an alternative to FW for inducing a molt in laying hens based on reduced nonnutritive pecking behavior, head movements, and greater feeding activity. PMID- 18420971 TI - The effect of quicklime (CaO) on litter condition and broiler performance. AB - High levels of phosphorus and pathogens in runoff are 2 major concerns following manure applications to fields. Phosphorus losses from fields following manure applications have been linked to the solubility of phosphorus in manure; therefore, by decreasing manure phosphorus solubility, a decrease in phosphorus loss in runoff should be apparent. The objective of this research was to develop a process using quicklime that would result in reduced phosphorus solubility and bacteria counts in broiler litter. The 4 litter treatments evaluated were T1, new wood shavings without the addition of quicklime; T2, used, untreated broiler litter; T3, used litter with 10% quicklime (based on the weight of the litter); and T4, used litter with 15% quicklime (based on the weight of the litter). Body weight, cumulative feed consumption, and feed conversion (feed:BW) were determined on a weekly basis through 42 d of age. Mortality was recorded daily. Carcass weights and percentages of carcass yield without giblets were determined prechill. Litter pH, total phosphorus, nitrogen, soluble phosphorus, litter moisture (%), and total plate counts were measured for each litter treatment on d 7 and 42 after bird placement. No significant differences were found for BW, feed consumption, feed conversion, mortality, carcass weight, or carcass yield. No breast or footpad blisters were observed. On d 7, 15% quicklime had higher (P < 0.001) pH (11.2) when compared with the other treatments. Percentages of phosphorus and nitrogen were lower (P < 0.001) for new wood shavings in comparison with the used litter treatments. Soluble phosphorus (ppm) was lower (P < 0.001) for 15% quicklime (2.75) when compared with new wood shavings (42.2), untreated broiler litter (439.2), and 10% quicklime (35.0). Although not significant, 15% quicklime had lower total plate counts (cfu/g) in comparison with the other treatments on d 1 and 10 postmixing and at 7 d after bird placement. Litter conditions on d 42 after bird placement were similar. We concluded that the use of quicklime as a treatment for broiler litter would initially reduce nitrogen and soluble phosphorus and bacteria counts without negatively affecting bird productivity. PMID- 18420972 TI - Bone mineral density and breaking strength of White Leghorns housed in conventional, modified, and commercially available colony battery cages. AB - Limited opportunity for movement and load-bearing exercise for conventionally caged laying hens leads to bone loss and increased susceptibility to osteoporosis, bone fractures, and cage layer fatigue, all of which compromise hen welfare and have negative consequences for production. The objective of this study was to compare bone mineral density (BMD) and strength measures of White Leghorns housed in conventional battery cages (CONV), cages modified to incorporate a nest box and perch (MOD), and commercially available, furnished colony cages with (CWDB) or without (CWODB) a raised dust bath. Hens reared on floor litter were randomly allocated to 1 of 4 cage systems at 19 wk of age. Hen day production and egg quality were measured between 20 and 64 wk. At 65 wk, hens were killed, and right femur, tibia, and humerus were excised. Bone mineral density was assessed using quantitative computed tomography, and breaking strength was measured with an Instron Materials Tester. In the femur and tibia, CONV hens exhibited lower total BMD, bone mass, cortical bone area, cortical bone mass, and bone-breaking strength than CWDB, CWODB, and MOD hens. Density and cross-sectional area of bone in the trabecular space was highest in CONV. In the humerus, total and cortical BMD and mass and breaking strength values were higher for colony-housed birds than hens in CONV and MOD. The MOD birds did not exhibit increased humeral BMD or strength measures over CONV hens. These findings provide evidence that hens housed in modified and colony cages, furnished systems that promote load-bearing movement, are better able to preserve cortical structural bone than conventionally caged hens and simultaneously have stronger bones. Furthermore, inclusion of raised amenities that encourage wing loading is necessary to reduce humeral cortical bone loss. The overall absence of correlation between egg production or quality and bone quality measures also suggests that improved bone quality in CWDB, CWODB, and MOD furnished cages is not the result of lowered egg production or quality. PMID- 18420973 TI - Development of methods for detection and quantification of avian influenza and Newcastle disease viruses in compost by real-time reverse transcription polymerase chain reaction and virus isolation. AB - Composting has been used for disposal of poultry carcasses and manure following outbreaks caused by avian influenza virus (AIV) and Newcastle disease virus (NDV), but methods are needed to test for survival of the viruses in compost to ensure biosecurity. Methods developed in the present study include extracting viruses from compost and purifying viral RNA. The extracted viruses were detected by virus isolation using embryonated chicken eggs, and the purified RNA was detected by real-time reverse transcription PCR (RRT-PCR). The virus isolation and the RRT-PCR methods were evaluated with 3 compost preparations that were produced from chicken manure mixed with corn silage, wood shavings, or wheat straw. The detection limits of both methods were 1,700 and 1,000 embryo lethal doses of AIV and NDV per gram of compost, respectively. The copy number of viral RNA quantified by RRT-PCR was highly correlated with the amount of virus in compost. The results suggested that the RRT-PCR method may be used as an alternative to the virus isolation method for rapid detection and accurate quantification of AIV and NDV in compost. PMID- 18420974 TI - Effects of dietary energy content on the performance of laying hens in furnished and conventional cages. AB - This study examined the effects of dietary energy content on the egg production and egg quality of hens kept in 3-hen conventional cages or 8-hen furnished cages. A total of 1,088 Lohmann Selected Leghorn hens were housed in either furnished or conventional cages and offered low- or high-energy diets (from 2,342 to 2,414 kcal/kg or from 2,581 to 2,629 kcal/kg) during 3 consecutive feeding phases of 20, 16, and 16 wk, respectively. The same dietary energy effects were observed in both cage systems. The hens that received the low-energy diet consumed more feed (P < 0.01) and produced fewer eggs per day (P < 0.05) than the birds fed the high-energy diet. Over the entire experiment, housing had no significant effects on production parameters, but during the third feeding phase, the production rate was smaller in furnished cages than in conventional cages (P < 0.01). Because of the greater live weight of the hens in furnished cages at the beginning of the experiment, these hens consumed more feed during the first feeding phase than the hens in conventional cages. During the third feeding phase, the hens in furnished cages consumed less feed than those in conventional cages (P < 0.01), probably because of their better feather cover. No differences in feed conversion ratio were found between the cage types. The results of this study confirm the results of previous studies providing evidence of equal production rates and feed conversion ratios in furnished and conventional cages. PMID- 18420975 TI - Effect of light intensity on live performance and processing characteristics of broilers. AB - This study investigated the effects of different light intensities provided via an increasing photoperiod program on broiler live performance and processing characteristics. A total of 1,080 male broilers were evenly distributed in 12 rooms. Six rooms were subjected to intensities of either 15 footcandles (FC) from 1 to 51 d (Bright), or 0.5 FC from 1 to 9 d and 0.1 FC from 9 to 51 d (Dim). Both intensity treatments were provided in an increasing photoperiod program (23L:1D, 1 to 9 d; 12L:12D, 9 to 16 d; 14L:10D, 16 to 23 d; 17L:7D, 23 to 30 d; 20L:4D, 30 to 37 d; and 23L:1D, 37 to 51 d). Feed consumption and BW were determined, and feed conversions were calculated approximately weekly. Mortalities were necropsied and recorded daily. At 51 d, 30 birds from each room were processed and cut up to determine weights and yields. Beginning at 23 and 30 d, respectively, BW and feed consumption were greater in the Dim treatment. At 51 d, Dim treatment BW was 4.7% greater and feed consumption was 3.9% greater. Feed conversion, metabolic and total mortality, and BW uniformity were not influenced by light intensity. Weights of lean carcass, total breast, fillets, tenders, and legs were from 4.9 to 6.2% greater in the Dim treatment, which was proportional to the BW difference and resulted in similar yields of these parts. However, wings were 9.9% heavier in the Dim treatment, which resulted in greater wing yield. Equal fat pad weights resulted in reduced fat pad yield in the Dim treatment. These results indicate that BW, feed consumption, and most parts weights were increased proportionally by providing 0.1 vs. 15 FC of light intensity via an increasing photoperiod program, and that only the yields of minor parts were affected by intensity. PMID- 18420976 TI - Genetic control of embryonic cardiac growth and functional maturation in turkeys. AB - Turkey experimental lines E (selected 44 yr for increased total egg production) and F (selected 38 yr for increased 16-wk BW) were mated reciprocally with the randombred control lines from which they were derived (RBC1 and RBC2, respectively), and the pureline and reciprocal cross poults were compared for their BW, heart weight, heart rates, myocardial glycogen and lactate concentrations, and plasma creatine kinase (CK) and lactate dehydrogenase (LDH) activities. The CK and LDH were used as indicators of cardiac insufficiency. Orthogonal contrasts of the data from the pureline and reciprocal cross data were used to estimate additive genetic effects, reciprocal effects (confounded maternal and sex-linked effects), and heterosis for each of the traits measured. Long-term selection for increased egg production in the E line has reduced embryo heart weight and has altered the energy metabolism of the myocardium. The differences in energy metabolism may be due to the more rapid heart rates. Conversely, long-term selection for increased 16-wk BW has significantly decreased the heart rate of F line embryos and has not changed the weight of the heart relative to the BW until the embryo has passed through the plateau stage. The F line embryos show a different energy metabolism that relies much more on gluconeogenesis. Embryo deaths occur more frequently in turkey embryos when the energy metabolism of the myocardium shows elevated glycogen to lactate ratios as it did in the pure E and F lines. PMID- 18420977 TI - Mixed model studies on inheritance of reproductive traits in laying hens - a Bayesian approach. AB - Segregation analyses performed for many livestock species indicate a mixed inheritance model of reproductive traits. Additionally, depending on the population, a given trait can be determined by a number of genes with large effects. Genetic backgrounds of hatch-ability and fertility in poultry are still not known sufficiently. The objectives of this study are to verify the hypothesis on segregation of single genes (1 vs. 2) affecting fertility and hatchability and to estimate a heritability of these traits. Records from 2,040 and 2,015 dams from full-pedigreed strains of Rhode Island Red (R33) and New Hampshire (N88) from a pedigree farm were analyzed. The percentage of fertilized eggs and the percentage of the eggs hatched of fertilized eggs were registered for dams only. Fertility was checked by candling on the eighth day of incubation. To obtain a binomial phenotypic scale, 10 eggs per dam were included into the analysis. Animal single-trait threshold models were used for the analysis of data. The first model included the effects of 2 single genes, 2 fixed effects of year and season, additive polygenic effects, and permanent environmental effects. In the second model, only 1 single gene effect was included. Additionally, the analysis based on the polygenic threshold model was also performed. The Gibbs sampling procedure was used. The significance of single gene effects was verified by highest posterior density regions. The obtained results clearly gave evidence for the segregation of 1 major gene for hatchability in strain R33. Furthermore, the mixed inheritance model can also be suggested for fertility in this strain. After the analysis, the polygenic heritabilities were very low (<0.11), whereas major polygenic heritability ranged from 0.05 to 0.12. PMID- 18420978 TI - Different behavioral and physiological responses in two genetic lines of laying hens after transportation. AB - Physiological and behavioral responses to transportation stress were examined in a chicken strain selected for high group productivity and survivability (HGPS) resulting from reduced cannibalism and flightiness in colony cages and in chickens from Dekalb XL (DXL) commercial strain. At 13 wk of age, 96 pullets per strain were randomly assigned to 4-bird cages within the same strain. At 17 wk of age, half of the cages of each strain (n = 12) were randomly used as experimental group. The birds of the experimental group were crated by line, with a caution to ensure all the birds in the same cage were unfamiliar to each other, and then transported for 2 h on a country road. After transportation, the birds were recaged in groups of 4 within the same line. Behavioral data were collected immediately after the recaging of the birds. Physical parameters (BW and organ weight), plasma corticosterone levels, blood and brain serotonin (5-HT) concentrations, and 5-HT1A receptor mRNA expressions were measured at 1 d posttransportation. Results showed that, compared with the control birds of each strain, transportation stress-induced behavioral changes in eating, drinking, and preening were found in the birds from both strains, but the HGPS birds showed a greater increase in drinking and preening (P < 0.01). In addition, after transportation stress, the HGPS birds had heavier adrenal glands (P < 0.01) with higher concentrations of plasma corticosterone (P < 0.01) and a trend to higher central 5-HT levels (P = 0.09) with a downregulated 5-HT1A receptor gene expression (P < 0.05), whereas the DXL birds had a higher H:L ratio (P < 0.05). The data indicate that there are a genetic basis of variations in chickens in response to transportation stress. The HGPS birds may have a better coping capability to transportation stress than DXL birds. PMID- 18420979 TI - Polymorphisms of vasoactive intestinal peptide receptor-1 gene and their genetic effects on broodiness in chickens. AB - Broodiness is a polygenic trait controlled by a small number of autosomal genes. Vasoactive intestinal peptide receptor-1 (VIPR-1) gene could be a candidate of chicken broodiness, and its genomic variations and genetic effects on chicken broodiness traits were analyzed in this study. The partial cloning and sequencing of the VIPR-1 gene showed that the average nucleotide diversity was 0.00669 +/- 0.00093 in Red Jungle Fowls (RJF), and 0.00582 +/- 0.00026 in domestic chickens. One hundred twenty-eight variation sites were identified in the 11,136-bp region of the chicken VIPR-1 gene. Twenty variation sites were genotyped using PCR-RFLP or PCR method to analyze average diversity, linkage-disequilibrium pattern, and haplotype structure in RJF, Xinghua chickens, Ningdu Sanhuang chickens, Baier Huang chickens, and Leghorn Layers. The RJF, Xinghua, Ningdu Sanhuang, and Baier Huang exhibited distinct characteristic of decreasing r(2) value over physical distance. Haplotype analyses showed that some variation sites of the 27-kb region from exon 6 to exon 11 could be associated with broodiness. The distribution of genotypic and allelic frequencies, and heterozygosities in the above 5 populations showed that A-284G, A+457G, C+598T, D+19820I, C+37454T, C+42913T, and C+53327T might be associated with broodiness. The 7 sites and the other 4 sites were genotyped in 644 NDH individuals under cage condition and were used for association analyses between each site and chicken broodiness traits. A significant association (P < 0.05) was found between C+598T in intron 2 and broody frequency (%). Another significant association (P < 0.05) was found between C+53327T and duration of broodiness, in which allele C was positive for DB. PMID- 18420980 TI - Growth rate of ascites-resistant versus ascites-susceptible broilers in commercial and experimental lines. AB - The high growth rate (GR) of contemporary broilers is driven by high rate of feed intake and metabolism. Because of the consequent high oxygen demand, especially when coupled with exposure to high altitude or low temperatures, some broilers fail to regulate oxygen supply and develop the ascites syndrome (AS), which leads to mortality and economic losses. Because of the association between high GR, oxygen demand, and AS, it has been suggested that AS is induced by high GR. If true, further GR enhancement should be avoided because it will increase the proportion of AS-susceptible individuals in contemporary stocks. An alternative hypothesis claims that AS is associated with high actual GR only because the latter increases oxygen demand and that there are genetically AS-resistant broilers that do not develop AS, even when exhibiting high GR. These hypotheses were tested in trials in the years 2002 and 2006, with broilers differing in potential GR: contemporary fast-growing commercial lines and an experimental line derived from commercial broilers in 1986, and (in 2002 only) divergently selected AS-susceptible and AS-resistant lines. A protocol of high-challenge ascites inducing conditions (AIC) from d 19 was used to distinguish between AS susceptible and AS-resistant individuals and to determine their GR up to this age. The difference in AS incidence between the divergent lines (93.9 vs. 9.5%) was not explained by the 5% difference in their GR, thus indicating a lack of genetic correlation. In the broiler lines, AS incidence was 31 and 47% in 2002 and 2006, respectively, and 32% in the 1986 slow-growing line. Most broilers that remained healthy under the high-challenge AIC exhibited the same early GR and BW as those that later developed AS. These results, and the relatively high incidence of AS in the slow-growing line, indicate that there is very little, if any, direct genetic association between AS and genetic differences in potential GR, and suggest that AS-resistant broilers can be selected for higher GR and remain healthy even under AIC. PMID- 18420981 TI - Additions of killed whole cell bacteria preparations to Freund complete adjuvant alter laying hen antibody response to soluble protein antigen. AB - Passive transfer of antibodies from hen to egg has value to both the producer of commercial polyclonal egg antibody and the producer of hatching eggs. Water-in oil emulsions are commonly amended with immune stimulants such as Mycobacteria (e.g., Freund complete adjuvant; FCA) to increase antibody production to soluble protein antigens (SPA). Recent discoveries of the mechanisms by which microbial products act as adjuvants led us to hypothesize that additions of killed whole cell bacteria (bacterins) to FCA could improve antibody responses to SPA. All injections used in each experiment were water-in-oil emulsions (50:50) containing 3 mg/mL of phospholipase A(2) (PLA(2)) immunogen. Additionally, all primary control and treatment injections contained heat-killed Mycobacterium butyricum immunogens from FCA. In addition to PLA(2) and FCA, primary treatment injections contained various microbial bacterin immunogens. Hence, the experimental treatment of all experiments was addition of a commercial source of microbial bacterin to FCA for the primary injection only. Booster injections were the same as the primary control injections except Freund incomplete adjuvant replaced FCA. Anti-body titers to PLA(2) in yolk were determined by ELISA. Bacterins tested as additives to FCA were Escherichia coli, Staphylococcus aureus, Streptococcus suis, and Corynebacterium pseudotuberculosis. Escherichia coli bacterin added to FCA decreased egg yolk antibody titer to SPA by 23% in hens of different ages and strains (P < 0.0001). In a second experiment, a 51% decrease in antibody production associated with E. coli bacterin was sustained for several weeks after the primary immunization (P = 0.003). Staphylococcus aureus or Streptococcus suis combined with FCA increased egg yolk antibody 62 and 51%, respectively (P < 0.05), and Corynebacterium pseudotuberculosis had no effect. In conclusion, the addition of bacterin to FCA can influence hen antibody response to SPA as measured in egg yolks. It is hypothesized that the difference in antibody production may be related to the composition of various pathogen associated molecular patterns in the primary injection. PMID- 18420982 TI - Decreased specific antibody responses to alpha-Gal-conjugated antigen in animals with preexisting high levels of natural antibodies binding alpha-Gal residues. AB - High levels of natural antibodies (NAb) binding the alpha-Gal residue (Galalpha1 3Galbeta1-4GlcNAc) are found in poultry (and humans), which is probably reflected by high levels of natural agglutinating antibodies (Ab) to rabbit red blood cells (RRBC) in plasma from chickens (and humans). Recently, it was shown that alpha Gal conjugation of proteins induced higher antiprotein Ab responses in alpha-Gal knockout mice, suggesting immune-enhancing features of preexisting Ab binding carbohydrate-protein conjugates. We challenged chickens s.c. with either alpha Gal-conjugated human serum albumin (HuSA), beta-Gal-conjugated HuSA, or unconjugated ("native") HuSA, respectively, and measured primary and secondary Ab responses to HuSA, including isotype IgM and IgG responses, and cellular immune responses in vitro (lymphoproliferation) to HuSA or concanavalin A. alpha-Gal conjugation, but not beta-Gal conjugation, of HuSA resulted in significantly decreased primary and secondary Ab responses to HuSA, especially IgG isotype responses, as compared with Ab responses to native HuSA. Lymphoproliferation in vitro was also decreased, although not significantly, in birds challenged with alpha-Gal-conjugated HuSA. High levels of agglutinating Ab levels to RRBC and NAb binding porcine thyroglobulin were detected in all birds, as was true for (natural) Ab levels binding alpha-Gal-conjugated HuSA before immunization, whereas low levels of preexisting (natural) antibodies directed to native HuSA were present in plasma before immunization. Levels of RRBC agglutinins and Ab binding thyroglobulin were not affected by immunization with HuSA, alpha-Gal conjugated HuSA, or beta-Gal-conjugated HuSA. Our data confirm the presence of high levels of (preexisting) NAb in the plasma of chickens directed to the alpha Gal residue. The decreased responsiveness to alpha-Gal-bearing antigens in the current study shows that, in addition to immune-enhancing features, NAb may also have suppressive effects on specific immune responses, which substantiates the regulatory role of innate immunity (NAb) in mounting specific immune responses. PMID- 18420983 TI - Detection of T lymphocytes in intestine of broiler chicks treated with Lactobacillus spp. and challenged with Salmonella enterica serovar Enteritidis. AB - The expression of immune response in the form of leukocytic infiltrate by CD3+, CD4+, and CD8+ cells in the epithelium and in the intestinal lamina propria of chicks was studied in the present work by means of immunohistochemical reaction. The chicks were treated with Lactobacillus spp. or cecal microflora (CM) and experimentally challenged or not with Salmonella enterica serovar Enteritidis. The 320 birds utilized were divided into 4 groups containing 80 chicks each and submitted to treatments with Lactobacillus reuteri, Lactobacillus salivarius, Lactobacillus acidophilus, and CM. Each group was subdivided into 4 subgroups of 20 birds each and classified into a subgroup that did not receive treatment (negative control), subgroup treated, subgroup treated and challenged with Salmonella Enteritidis, and subgroup only challenged with Salmonella Enteritidis (positive control). The results obtained show that the treatment with L. reuteri, L. salivarius, L. acidophilus, or CM and challenged or not with Salmonella Enteritidis determine immune response in the form of leukocytic infiltrate by CD3+ and CD8+ lymphocytes followed by CD4+ in the epithelium and in the lamina propria of the duodenum, jejunum, and cecum of chicks up to 12 d of age. The quantity of CD3+ lymphocytes was significantly higher (P < 0.05) in the intestine of chicks treated with L. acidophilus or CM and challenged or not with Salmonella Enteritidis; however, the higher quantity of CD8+ lymphocytes was in the intestine of chicks treated with CM and challenged with Salmonella Enteritidis. The duodenum was the segment in which the immune response by T cells (CD3+, CD4+, and CD8+) was stimulated with the greatest intensity, followed by, respectively, the jejunum and cecum. The quantity of CD3+ lymphocytes present in the duodenum, jejunum, and cecum increases with the age of chicks, independent of the stimulus determined by treatments or challenge. PMID- 18420984 TI - The effect of carrier for vitamin E on liver concentrations of vitamin E and vitamin E excretion in broilers. AB - Two experiments were conducted to determine the effect of 2 carrier sources of vitamin E on growth performance and excreta and liver vitamin E concentrations of broilers. Chicks were pretested from d 0 to 7 posthatching on a corn-soybean meal diet without vitamin E supplementation, and the experiments lasted from d 7 to 19 posthatching. Each treatment in both experiments was replicated with 6 pens with 5 chicks each. Initial and final BW were 155 and 684 g in experiment 1 and 155 and 691 g in experiment 2. In experiment 1, the dietary treatments were the corn soybean meal diet with varying concentrations (0, 30, 100, or 300 IU/kg) of supplemental vitamin E either absorbed to verxite (VE) or adsorbed to silica (SE). In experiment 2, the dietary treatments were the 0 and 30 IU/kg of vitamin E as VE or SE. In experiment 1, feed intake decreased and gain:feed increased as concentration of VE increased, but feed intake increased and gain:feed decreased as concentration of SE increased (source x concentration, P < 0.03). Gain, feed intake, and gain:feed were not affected (P > 0.10) by the vitamin E carrier source in experiment 2. The mean excreta vitamin E concentration at d 7 posthatching was 17.2 IU/kg (DM basis). The percentages of vitamin E excreted were based on analyzed vitamin E concentrations in the diet. At 100 and 300 IU/kg of supplemental vitamin E, an average of 94 and 44% of vitamin E intake from broilers fed vitamin E from VE and SE, respectively, was excreted (vitamin E source, P < 0.01; source x concentration, P < 0.08), but at 30 IU/kg of vitamin E, 49 and 45% of vitamin E intake from broilers fed vitamin E from VE and SE was excreted. In experiment 2, 52 and 43% of vitamin E intake from VE and SE was excreted (source, P < 0.02). Liver alpha-tocopherol concentration was not different (P > 0.10) between the sources of vitamin E in either experiment. Increased concentrations of vitamin E increased liver alpha-tocopherol concentrations (P < 0.01). On the basis of the results of excreted vitamin E, vitamin E adsorbed to SE was more available than vitamin E absorbed to VE, but on the basis of liver vitamin E concentration, their availabilities were similar. PMID- 18420986 TI - The effect of dietary phosphorus level and phytase supplementation on growth performance, bone-breaking strength, and litter phosphorus concentration in broilers. AB - This research was conducted to determine the effects of feeding different P levels with and without phytase supplementation on broiler growth performance, bone-breaking strength (BBS), and litter P concentration. An experiment with 4 trials was conducted with 7,840 Ross x Ross straight-run broilers. For each trial, 1,960 broilers were allotted on d 0 to treatments, with 7 replications each and with 70 broilers per replication. The broilers were fed a 4-period feeding program consisting of starter (0 to 14 d), grower (14 to 32 d), finisher (32 to 41 d), and withdrawal (41 to 50 d) periods. For each trial, the same pen was used continuously for each treatment-replication combination, and the litter was not removed between trials. Broilers were fed a control diet [0.43, 0.40, 0.36, or 0.32% nonphytate P (nPP)] in the starter, grower, finisher, and withdrawal periods, respectively, a low Ca and P (LCaP) diet with a 0.05% reduction in nPP in each period, and these 2 diets supplemented with phytase at 600 phytase units/kg (nPP and Ca were reduced by 0.094% in diets with phytase). Diet did not affect (P > 0.10) broiler performance in the starter or withdrawal periods. Generally, both phytase addition and the LCaP diet decreased some aspects of growth performance during the grower and finisher periods. There was no main effect of phytase on BBS, but BBS was decreased in the broilers fed the LCaP diet with phytase addition (nPP x phytase, P < 0.01) in the grower period, and BBS was decreased in the finisher (P < 0.02) and withdrawal (P < 0.01) periods for broilers fed the LCaP diet. Total P, soluble P (SP), and reactive soluble P (RSP) were decreased (P < 0.04) in the litter of broilers fed the LCaP diets. Total P, SP, and RSP were decreased (P < 0.01) in the litter of broilers fed phytase. These data indicate that phytase supplementation at 600 phytase units/kg reduces growth in some periods, has no effect on BBS, and reduces total P, SP, and RSP in the litter. PMID- 18420985 TI - Influence of enzyme supplementation and heat processing of barley on digestive traits and productive performance of broilers. AB - We studied the influence of enzyme supplementation (ES) of the diet and heat processing (HP) of barley on digestive traits and productive performance of broilers from 1 to 42 d of age. There were 6 diets arranged factorially with 2 doses (0 and 500 ppm) of a fungal enzyme complex with beta-glucanase and xylanase activity and 3 HP of barley (raw, micronized, and expanded). In addition, a control diet based on raw corn without ES was also included from 1 to 21 d of age. Enzymes reduced intestinal viscosity (IV) at all ages (P < or = 0.001) and water intake at 21 d of age (P < or = 0.01) and increased DM of the ileal contents at 28 d (P < or = 0.001). Also, ES increased total tract apparent retention of nutrients and BW gain and feed conversion ratio from 1 to 42 d of age (P < or = 0.001). Heat processing of barley increased IV at 7 and at 28 d of age, and DM of ileal contents (P < or = 0.05) at 28 d of age. In addition, HP improved feed intake (P < or = 0.01) and BW gain (P < or = 0.001) from 1 to 7 d of age, but the effects disappeared after 21 d of age. From 1 to 7 d of age, chicks fed micronized barley had higher IV, gained less weight, and had poorer feed conversion ratio than chicks fed expanded barley (P < or = 0.05). It is concluded that barley with enzymes can substitute for all of the corn in diets fed to broilers from 1 to 21 d of age. Enzymes improved digestive traits, retention of nutrients, and broiler performance from 1 to 42 d of age, and HP of barley improved performance from 1 to 7 d of age. The effects of HP of barley on broiler performance were more evident with expansion than with micronization. PMID- 18420987 TI - The effect of dietary sinapic acid (4-hydroxy-3, 5-dimethoxy-cinnamic acid) on gastrointestinal tract microbial fermentation, nutrient utilization, and egg quality in laying hens. AB - Plant-based simple phenolics are natural antimicrobial and antioxidant compounds that may possess biological activity when included in animal diets. Previous work in our lab has shown that sinapic acid (SA) found in canola meal improves nutrient retention and alters gut microbiota when fed at low levels (0 to 0.10%) to broiler chickens. It was of interest to investigate the effect of dietary SA in laying hens that possess a more mature intestinal microbiota. Therefore, 2 experiments were conducted to determine the effects of dietary SA on performance, nutrient digestibility, and egg quality in laying hens. Sinapic acid (Sigma Aldrich, 98% purity) was included in diets at 0, 0.025, 0.050, and 0.075% in experiment 1, and 0 and 0.5% in experiment 2. Both experiments were conducted for 21 d. Dietary treatment did not affect production traits and gastrointestinal tract volatile fatty acid production in either experiment. Apparent metabolizable energy and protein digestibility were also unaffected by treatment in experiment 1, but in experiment 2, feeding 0.5% SA increased both AME(n) and protein digestibility in comparison to the unsupplemented control. Egg weight, albumen weight, yolk weight, egg specific gravity, shell thickness, and Haugh units were not significantly affected by dietary treatments in either of the experiments. Sinapic acid was not detected in eggs from experiment 1 but in experiment 2, low levels of SA were detected in both the yolk and albumen of eggs from hens fed 0.5% SA. Further research is needed to more completely define the relationship between dietary SA level and nutrient utilization as well as the accumulation of SA in egg components. PMID- 18420988 TI - The effect of gender on the bacterial community in the gastrointestinal tract of broilers. AB - The effect of gender on the growth, carcass yield, and nutritional requirements of chickens has been well documented, but little is known about how the sex of a chicken affects the bacterial population of the gastrointestinal tract. Therefore, our objective was to evaluate the biodiversity of the bacterial community in the gastrointestinal tract of male and female broilers. An experiment was conducted with Cobb 500 broiler chicks that were vent sexed at 0 d of age and allocated to 8 pens of 25 chicks per gender. All birds were fed a nonmedicated corn-soybean meal starter diet from 0 to 21 d of age. At 3, 7, 14, and 21 d of age, chicks were randomly selected and ileums were taken for bacterial sampling. Bacterial DNA was isolated from the digesta of the ileum, and denaturing gradient gel electrophoresis was used to examine PCR-amplified fragments of 16S ribosomal DNA. Denaturing gradient gel electrophoresis analyses revealed that the bacterial communities separated into 2 gender-specific groups, with less than 30% similarity between populations. Furthermore, as the birds aged, the similarity of the intestinal bacterial community decreased within each gender. Although ileal bacterial population differences within and between genders were noted as early as d 3, differences in growth rate between males and females were not noted until d 21 (data not shown). This suggested that non growth-related factors influenced the composition of intestinal bacterial communities. PMID- 18420989 TI - Temporal distribution and genetic fingerprinting of Salmonella in broiler flocks from southern Japan. AB - During the 1998 to 2003 period, cecal contents of 4,024 broiler chickens from 252 flocks raised in 63 holdings were examined for Salmonella. The aims were to establish the actual status of the infection, its temporal distribution, prevalent serotype, and common genotype among broiler flocks brought at the slaughterhouse. Collected samples were preenriched in Hajna tetrathionate broth, and after 24 h of incubation, 10 microL of the broth was streaked on selective Rambach agar plate. Suspected scarlet color colonies of Salmonella were cloned on nutrient agar, confirmed through biochemical tests and sero-typed using O and H antigens. Pulsed field gel electrophoresis technique generated DNA fragments banding patterns and established their clonal relatedness. Salmonella was isolated from 563 (14%) samples in 179 (71%) flocks. The flock situation varied from Salmonella-negative holdings (n = 9), positive-flocks from persistently infected holdings (n = 21), and holdings (n = 19) that showed fluctuations with alternating negative and positive flocks for variable time periods. Fourteen holdings (negative, n = 5 and positive, n = 9) were sampled once throughout the study period. Seasonality component was not observed, and salmonellae were found colonizing broiler ceca in warm and cold months. Predominant serovar was Salmonella Infantis (93.3%; n = 525). Macrorestriction fingerprints of Salmonella Infantis using XbaI presumed the isolates to be derived from a common parent. Enhanced discrimination by BlnI digestion produced 3 banding patterns that were closely related genetically and hence epidemiologically related. Such epidemiological information may enable producers to formulate effective control action plan tailored for individual holdings with special emphasis on biosecurity, hygiene, and pest control. PMID- 18420990 TI - Effects of cold stress on the messenger ribonucleic acid levels of corticotrophin releasing hormone and thyrotropin-releasing hormone in hypothalami of broilers. AB - In this study, seventy 1-d-old male broiler chicks were randomly allocated to 10 groups to investigate the effect of cold stress on the messenger RNA (mRNA) levels of corticotrophin-releasing hormone (CRH) and thyrotropin-releasing hormone (TRH) in hypothalami. The chicks were maintained at 30 +/- 2 degrees C temperature and given free access to standard chow and water. Until 15 d old, the 6 treatment groups were maintained at 12 +/- 1 degrees C. Hypothalami were collected for the assessment of the mRNA levels by semiquantitative reverse transcription-PCR after stress termination. Cold stress significantly decreased the mRNA levels of CRH in 6 and 12 h treatment groups and significantly increased the mRNA levels of TRH in 1, 6, and 12 h treatment groups during acute cold stress. There were no significant differences in the mRNA levels of CRH and TRH among different control groups during chronic cold stress. However, chronic cold stress resulted in a significant increase of the mRNA levels of CRH and a significant decrease of the mRNA levels of TRH compared with the corresponding control groups. The results indicate that the 2 genes show different response to cold stress at the mRNA levels, and on the other hand, the different degree of cold stress also produces different effects on the identical gene. PMID- 18420991 TI - Expression of avian beta-defensins in the oviduct and effects of lipopolysaccharide on their expression in the vagina of hens. AB - The aims of this study were to (i) determine the types of avian beta-defensin genes (AvbetaD) expressed in the hen oviduct and (ii) to examine the effects of lipopolysaccharide (LPS) treatment in vivo on their expression in the vagina. Birds were i.v. treated with LPS (1 mg/kg of BW), and subsequently the oviducts were analyzed 0, 3, 6, 12, or 24 h after LPS administration. The mRNA expression for AvbetaD was examined by reverse transcription-PCR using RNA preparations from the mucosal tissues of all the oviductal segments. Furthermore, changes in their mRNA expression profiles in the vagina were analyzed by semiquantitative reverse transcription-PCR. The AvbetaD-1, -2, -3, -4, -5, -7, -8, -9, -10, -11, and -12 were identified in each oviductal segment from infundibulum to vagina. Among these AvbetaD, the expression of AvbetaD-3, -5, -10, -11, and -12 in the vagina were significantly increased in response to LPS treatment, whereas the others did not show significant changes. These results suggest that all 11 types of AvbetaD are expressed in the hen oviduct and at least 5 of them in the vagina show increased expression in response to LPS. PMID- 18420992 TI - Reduction of Salmonella Enteritidis in shell eggs using directional microwave technology. AB - Microwaves have been shown to cause thermal as well as nonthermal destruction of pathogens such as Salmonella Enteritidis, which is commonly found in shell eggs. The objective of this study was to examine the use of new directional microwave technology to reduce Salmonella Enteritidis without causing any detrimental effects on quality in white and brown eggs. Treatments were control and microwaved white and brown eggs. Applying directional microwave technology resulted in a 2-log reduction of Salmonella Enteritidis in both the high (10(5) cfu/g) and low (10(2) cfu/g) inoculum. At d 0, there were no differences in water activities, albumen pH, and combined pH between treatments; however, there were significant changes in yolk pH. Collectively, these results indicate that applying directional microwave technology can reduce Salmonella Enteritidis in shell eggs without causing any detrimental effects to quality. PMID- 18420993 TI - The effect of chilling in cold air or ice water on the microbiological quality of broiler carcasses and the population of Campylobacter. AB - Cold air or ice water can be used to chill poultry carcasses after slaughter. The objective of this study was to compare the effect of 2 chill methods on broiler carcass bacteria. Broiler carcasses were cut in half along the dorsal-ventral midline; one half was subjected to an ice-water immersion chill in an agitated bath for 50 min, whereas the reciprocal half was subjected to an air chill in a 1 degrees C cold room for 150 min. Total aerobic bacteria, coliforms, Escherichia coli, and Campylobacter were enumerated from half-carcass rinses. Species of Campylobacter isolates was determined by a commercial PCR method, which was followed by molecular subtyping with pulsed-field gel electrophoresis and determination of antimicrobial susceptibility to 9 drugs. Although significantly fewer of each bacterial type were detected per milliliter from immersion-chilled carcasses than from air-chilled carcasses, in each case the difference was less than 1 log(10) cfu/mL. Chilling method did not affect species; both Campylobacter jejuni and Campylobacter coli were recovered. Results of pulsed-field gel electrophoresis subtyping did not suggest that either chilling method selected for any specific subtypes; most subtypes were found on carcass halves used for both the air chill and water immersion chill. Resistance to 2 antimicrobial drugs was noted in 9 C. coli isolates, 6 from air-chilled carcass halves and 3 from immersion-chilled carcass halves. These data showed that immersion-chilled carcasses had lower numbers of bacteria; however, the difference was not large and may have been due to simple dilution. Both methods were effective for lowering carcass temperature, and neither chilling method seemed to select for specific species, subtypes, or antimicrobial-resistant Campylobacter. PMID- 18420994 TI - SIRT1, a longevity gene, downregulates angiotensin II type 1 receptor expression in vascular smooth muscle cells. AB - OBJECTIVE: Resveratrol (3,5,4'-trihydroxystilbene), a polyphenol found in red wine, is known to activate sirtuin1 (SIRT1), a longevity gene. Previous studies have demonstrated that resveratrol extends the life span of diverse species through activation of SIRT1. It was also reported that inhibition of angiotensin II function by angiotensin II type I receptor (AT1R) antagonist prolonged rat life span. We, therefore, hypothesized that resveratrol may inhibit the renin angiontein system and examined whether resveratrol affects AT1R expression in vascular smooth muscle cells (VSMCs). METHODS AND RESULTS: Northern and Western blot analysis revealed that resveratrol significantly decreased the expression of AT1R at mRNA and protein levels in a dose- and time-dependent manner. Overexpression of SIRT1 reduced AT1R expression whereas nicotinamide, an inhibitor of SIRT1, increased AT1R expression and reversed the resveratrol induced AT1R downregulation. AT1R gene promoter activity was decreased by resveratrol, but resveratrol did not affect the AT1R mRNA stability. Deletion analysis showed that the most proximal region of AT1R gene promoter containing Sp1 site is responsible for downregulation. Administration of resveratrol suppressed AT1R expression in the mouse aorta and blunted angiotensin II-induced hypertension. CONCLUSIONS: Resveratrol suppressed AT1R expression through SIRT1 activation both in vivo and in vitro. The inhibition of the renin-angiotensin system may contribute, at least in part, to the resveratrol-induced longevity and antiatherogenic effect of resveratrol. PMID- 18420995 TI - Temporally expressed PDGF and FGF-2 regulate embryonic coronary artery formation and growth. AB - OBJECTIVE: PDGF and FGF-2 are important regulators of vascular wall assembly. We tested the hypothesis that their embryonic temporal expression facilitates 2 specific events: (1) the endothelial invasion of the aortic root to form the coronary artery stems and (2) the subsequent growth and development of the arterial tree. METHODS AND RESULTS: Addition of FGF-2 and PDGF-BB proteins to embryonic quail heart explants stimulated a 3- and 7-fold increase, respectively, in tubulogenesis, whereas neutralizing antibodies to these growth factors attenuated tubulogenesis by 40%. Anti-FGF-2 and anti-PDGF neutralizing antibodies were then introduced in ovo via the vitelline vein at various embryonic (E) days. When injections occurred before coronary ostial formation, the embryos usually developed only 1 coronary artery or lacked coronary arteries. When 1 or both major coronary arteries formed: (1) their branches had a thinner tunica media, and (2) smooth muscle investment did not progress as far distally as in shams. Other anomalies included smaller diameter coronary artery stems in some hearts. Inhibition of VEGF via injections of aflibercept (VEGF-Trap, a VEGFR-1 and -2 chimera), previously shown to be essential for coronary stem formation, limited development of the coronary arteries even though introduced after formation of coronary ostia (at E9 or EI0). CONCLUSIONS: Our data (1) document a role for FGF 2 and PDGF in the temporal regulation of coronary artery stem formation and growth of the coronary arterial tree and (2) reveal that VEGF expression is required for normal artery/arterial formation, even after coronary artery stem formation. PMID- 18420997 TI - Cholesterol absorption from the intestine is a major determinant of reverse cholesterol transport from peripheral tissue macrophages. AB - OBJECTIVE: We examined the effect of ezetimibe, a cholesterol absorption (CA) inhibitor, and genetic determinants of CA on reverse cholesterol transport (RCT) from subcutaneously injected macrophages using a new dual isotope label technique. METHODS AND RESULTS: Treatment of C57BL/6J mice with ezetimibe decreased dietary CA by 86% and increased RCT from peripheral tissue macrophages (PTM) by 6-fold (P<0.0001). Moreover, congenic 14DKK mice with a modest 41% decrease in dietary CA displayed a 67% increase in RCT from PTM (P<0.007). CONCLUSIONS: These findings indicate that pharmacological and genetic modifiers of cholesterol absorption are major determinants of reverse cholesterol transport from peripheral tissue macrophages. PMID- 18420996 TI - Role of smooth muscle cGMP/cGKI signaling in murine vascular restenosis. AB - BACKGROUND: Nitric oxide (NO) is of crucial importance for smooth muscle cell (SMC) function and exerts numerous, and sometimes opposing, effects on vascular restenosis. Although cGMP-dependent protein kinase type I (cGKI) is a principal effector of NO, the molecular pathway of vascular NO signaling in restenosis is unclear. The purpose of this study was to examine the functional role of the smooth muscle cGMP/cGKI signaling cascade in restenosis of vessels. METHODS AND RESULTS: Tissue-specific mouse mutants were generated in which the cGKI protein was ablated in SMCs. We investigated whether the absence of cGKI in SMCs would affect vascular remodeling after carotid ligation or removal of the endothelium. No differences were detected between the tissue-specific cGKI mutants and control mice at different time points after vascular injury on a normolipidemic or apoE deficient background. In line with these results, chronic drug treatment of injured control mice with the phosphodiesterase-5 inhibitor sildenafil elevated cGMP levels but had no influence on the ligation-induced remodeling. CONCLUSIONS: The genetic and pharmacological manipulation of the cGMP/cGKI signaling indicates that this pathway is not involved in the protective effects of NO, suggesting that NO affects vascular remodeling during restenosis via alternative mechanisms. PMID- 18420998 TI - CD44 expressed on both bone marrow-derived and non-bone marrow-derived cells promotes atherogenesis in ApoE-deficient mice. AB - OBJECTIVE: The purpose of this study was to distinguish the contributions of CD44 expressed on bone marrow-derived and non-bone marrow-derived cells to atherosclerosis. METHODS AND RESULTS: Using bone marrow chimeras, we compared the contributions of CD44 expressed on bone marrow-derived cells versus non-bone marrow-derived cells to the vascular inflammation underlying atherosclerosis. We show that CD44 in both bone marrow-derived and non-bone marrow-derived compartments promotes atherosclerosis in apoE-/- mice and mediates macrophage and T cell recruitment to lesions in vivo. We also demonstrate that CD44 on endothelial cells (ECs) as well as on macrophages and T cells enhances leukocyte endothelial cell adhesion and transendothelial migration in vitro. Furthermore, CD44 on vascular smooth muscle cells (VSMCs) regulates their hyaluronan (HA) dependent migration. Interestingly, in mice lacking CD44 in both compartments, where we observed the least inflammation, we also observed enhanced fibrous cap formation. CONCLUSIONS: CD44 expressed on bone marrow-derived and non-bone marrow derived cells both promote atherosclerosis in apoE-deficient mice. Furthermore, CD44 plays a pivotal role in determining the balance between inflammation and fibrosis in atherosclerotic lesions which can impact clinical outcome in humans. PMID- 18420999 TI - T-lymphocyte infiltration in visceral adipose tissue: a primary event in adipose tissue inflammation and the development of obesity-mediated insulin resistance. AB - BACKGROUND: Adipose tissue inflammation may play a critical role in the pathogenesis of insulin resistance (IR). The present study examined the role of lymphocytes in adipose tissue inflammation and IR. METHODS AND RESULTS: In a mouse model of obesity-mediated IR, high-fat diet (HFD) induced IR already after 5 weeks, which was associated with a marked T-lymphocyte infiltration in visceral adipose tissue. In contrast, recruitment of macrophages was delayed with an increase of MAC3-positive staining and F4/80 mRNA expression after 10 weeks of HFD, suggesting a dissociation of macrophage invasion into adipose tissue and IR initiation. In patients with type 2 diabetes, lymphocyte content in adipose tissue biopsies significantly correlated with waist circumference, a marker of IR. Immunohistochemical staining of human adipose tissue revealed the presence of mainly CD4-positive lymphocytes as well as macrophage infiltration. Most macrophages were HLA-DR-positive, reflecting activation through IFNgamma, a cytokine released from CD4-positive lymphocytes. CONCLUSIONS: Proinflammatory T lymphocytes are present in visceral adipose tissue and may contribute to local inflammatory cell activation before the appearance of macrophages, suggesting that these cells could play an important role in the initiation and perpetuation of adipose tissue inflammation as well as the development of IR. PMID- 18421000 TI - Acute elevation of plasma PLTP activity strongly increases pre-existing atherosclerosis. AB - OBJECTIVE: A transgenic mouse model was generated that allows conditional expression of human PLTP, based on the tetracycline-responsive gene system, to study the effects of an acute increase in plasma PLTP activity as may occur in inflammation. METHODS AND RESULTS: The effects of an acute elevation of plasma PLTP activity on the metabolism of apolipoprotein B-containing lipoproteins and on diet-induced pre-existing atherosclerosis were determined in mice displaying a humanized lipoprotein profile (low-density lipoprotein receptor knockout background). Induced expression of PLTP strongly increases plasma VLDL levels in LDL receptor knockout mice, whereas VLDL secretion is not affected. The elevation in plasma triglyceride levels is explained by a PLTP-dependent inhibition of VLDL catabolism, which is caused, at least partly, by a decreased lipoprotein lipase activity. Together with the decreased plasma HDL levels, the acutely increased PLTP expression results in a highly atherogenic lipoprotein profile. Induction of PLTP expression leads to a further increase in size of pre-existing atherosclerotic lesions, even on a chow diet. In addition, the lesions contain more macrophages and less collagen relative to controls, suggesting a less stable lesion phenotype. CONCLUSIONS: In conclusion, acute elevation of PLTP activity destabilizes atherosclerotic lesions and aggravates pre-existing atherosclerosis. PMID- 18421001 TI - Distension of the carotid artery and risk of coronary events: the three-city study. AB - OBJECTIVE: Arterial mechanical properties are of growing interest in the understanding of cardiovascular disease development. We aimed to determine the predictive value of carotid wall mechanics on coronary heart disease (CHD) in the Three-City study. METHODS AND RESULTS: At baseline, 3337 participants aged > or =65 years underwent a carotid B-mode ultrasonography. During a median follow-up of 43.4 months, 128 CHD occurred. Increased carotid distension (relative stroke change in lumen diameter) was significantly associated with CHD risk. Comparison of subjects in tertile 3 versus those in tertile 1 (reference) showed a hazard ratio (HR) of 1.80 (95% CI, 1.17 to 2.75). Controlling for various confounders including age, heart rate, brachial (or carotid) pulse pressure, and common carotid intima-media thickness did not alter the association between carotid distension and CHD with a HR of 1.79 (95% CI, 1.12 to 2.86; tertile 3 versus tertile 1). Brachial and carotid pulse pressures were also independently associated with CHD. No association was found between CHD and carotid distensibility coefficient, cross-sectional compliance coefficient, Young's elastic modulus, or beta stiffness index. CONCLUSIONS: In the elderly, increased carotid distension was independently predictive of CHD. This simple and noninvasive parameter might be of particular interest for cardiovascular risk assessment. PMID- 18421003 TI - Nanomedicine captures cardiovascular disease. PMID- 18421004 TI - The adult epicardium: realizing the potential for neovascular therapy. PMID- 18421005 TI - Cysteinyl-leukotrienes in cerebrovascular disease: angels and demons? PMID- 18421006 TI - Expanding the concept of telomere dysfunction in cardiovascular disease. PMID- 18421007 TI - HDL, PTX3, and vascular protection. PMID- 18421008 TI - Prokineticin receptor-1 induces neovascularization and epicardial-derived progenitor cell differentiation. AB - OBJECTIVE: Identification of novel factors that contribute to myocardial repair and collateral vessel growth hold promise for treatment of heart diseases. We have shown that transient prokineticin receptor-1 (PKR1) gene transfer protects the heart against myocardial infarction in a mouse model. Here, we investigated the role of excessive PKR1 signaling in heart. METHODS AND RESULTS: Transgenic mice overexpressing PKR1 in cardiomyocytes displayed no spontaneous abnormalities in cardiomyocytes but showed an increased number of epicardial-derived progenitor cells (EPDCs), capillary density, and coronary arterioles. Coculturing EPDCs with H9c2 cardiomyoblasts overexpressing PKR1 promotes EPDC differentiation into endothelial and smooth muscle cells, mimicking our transgenic model. Overexpressing PKR1 in H9c2 cardiomyoblasts or in transgenic hearts upregulated prokineticin-2 levels. Exogenous prokineticin-2 induces significant outgrowth from neonatal and adult epicardial explants, promoting EPDC differentiation. These prokineticin-2 effects were abolished in cardiac explants from mice with PKR1-null mutation. Reduced capillary density and prokineticin-2 levels in PKR1 null mutant hearts supports the hypothesis of an autocrine/paracrine loop between PKR1 and prokineticin-2. CONCLUSIONS: Cardiomyocyte-PKR1 signaling upregulates its own ligand prokineticin-2 that acts as a paracrine factor, triggering EPDCs proliferation/differentiation. This study provides a novel insight for possible therapeutic strategies aiming at restoring pluripotency of adult EPDCs to promote neovasculogenesis by induction of cardiomyocyte PKR1 signaling. PMID- 18421009 TI - Proteomics identification of cyclophilin a as a potential prognostic factor and therapeutic target in endometrial carcinoma. AB - Endometrial carcinoma is one of the most common malignancies of the female genital tract, and there is an urgent need for discovery of novel factors for prognostic assessment and therapeutic targets to endometrial carcinoma. Herein a two-dimensional gel electrophoresis and MALDI-Q-TOF MS/MS-based proteomics approach was used to identify differentially expressed proteins in endometrial carcinoma. Of the 99 proteins identified, cyclophilin A was one of the most significantly altered proteins, and its overexpression was confirmed using RT-PCR and Western blot analyses. Immunohistochemistry suggested a link between cyclophilin A expression and poor differentiation and decreased survival (p < 0.01). Knockdown of cyclophilin A expression by RNA interference led to the significant suppression of the cell growth and the induction of apoptosis in endometrial carcinoma HEC-1-B cells in vitro (p < 0.01) and the inhibition of tumor growth in vivo (p < 0.01). These data suggest that cyclophilin A may serve as a novel prognostic factor and possibly an attractive therapeutic target for endometrial carcinoma. PMID- 18421010 TI - Patient care and management of frontotemporal lobar degeneration. AB - Frontotemporal lobar degeneration (FTLD) is a neurodegenerative disease that affects frontal and temporal regions of the brain. Two proteins indicated in the pathology are tau and the recently discovered TDP-43. Major manifestations include progressive aphasia and a disorder of social comportment. The diagnosis of a patient includes a detailed cognitive exam, clinical testing, and neuroimaging techniques. The current goal of therapy for FTLD is symptomatic management with medications borrowed from other conditions. Nonpharmacologic management such as behavioral interventions and environmental engineering are also efficacious in optimizing quality of life. PMID- 18421011 TI - Strategies to support recruitment and retention of First Nations youth in baccalaureate nursing programs in Saskatchewan, Canada. AB - Aboriginal youth is one of the fastest growing of all populations in Saskatchewan today. This is a prime group to target for training in the health care professions. The need for strategies to support recruitment and retention in these programs is critical for maintaining our present standard and increasing demands of quality health care. Program initiatives and supports need to be implemented to encourage this population to enroll in and complete health care programs. Although only 5 years old, the University of Saskatchewan, First Nations University of Canada, and Saskatchewan Institute of Applied Science and Technology (SIAST) have created a viable northern nursing program with a retention rate of Aboriginal postsecondary students 13% greater than the provincial norm. They graduated their first class of nursing students from and for the North, May 2006. PMID- 18421012 TI - Pulmonary mastocytosis and enhanced lung inflammation in mice heterozygous null for the Foxf1 gene. AB - The Forkhead Box f1 (Foxf1) transcriptional factor (previously known as HFH-8 or Freac-1) is expressed in endothelial and smooth muscle cells in the embryonic and adult lung. To assess effects of Foxf1 during lung injury, we used CCl(4) and butylated hydroxytoluene (BHT) injury models. Foxf1(+/-) mice developed severe airway obstruction and bronchial edema, associated with increased numbers of pulmonary mast cells and increased mast cell degranulation after injury. Pulmonary inflammation in Foxf1(+/-) mice was associated with diminished expression of Foxf1, increased mast cell tryptase, and increased expression of CXCL12, the latter being essential for mast cell migration and chemotaxis. After ovalbumin (OVA) sensitization and OVA challenge, increased lung inflammation, airway hyperresponsiveness to methacholine, and elevated expression of CXCL12 were observed in Foxf1(+/-) mice. During lung development, Foxf1(+/-) embryos displayed a marked increase in pulmonary mast cells before birth, and this was associated with increased CXCL12 levels in the lung. Expression of a doxycycline inducible Foxf1 dominant-negative transgene in primary cultures of lung endothelial cells increased CXCL12 expression in vitro. Foxf1 haploinsufficiency caused pulmonary mastocytosis and enhanced pulmonary inflammation after chemically induced or allergen-mediated lung injury, indicating an important role for Foxf1 in the pathogenesis of pulmonary inflammatory responses. PMID- 18421013 TI - Prostaglandin E2 inhibits specific lung fibroblast functions via selective actions of PKA and Epac-1. AB - Via their capacities for proliferation and synthesis of matrix proteins such as collagen, fibroblasts are key effectors in the pathogenesis of fibrotic disorders such as idiopathic pulmonary fibrosis. Prostaglandin E(2) (PGE(2)) potently inhibits these functions in lung fibroblasts through receptor ligation and production of the second messenger cAMP, but the downstream pathways mediating such actions have not been fully characterized. We sought to investigate the roles of the cAMP effectors protein kinase A (PKA) and exchange protein activated by cAMP-1 (Epac-1) in modulating these two functions in primary human fetal lung IMR-90 fibroblasts. The specific roles of these two effector pathways were examined by treating cells with PKA-specific (6-bnz-cAMP) and Epac-specific (8 pCPT-2'-O-Me-cAMP) agonists, inhibiting PKA with the inhibitor KT 5720, overexpressing the PKA catalytic subunit, and silencing Epac-1 using short hairpin RNA. PGE(2) inhibition of collagen I expression was mediated exclusively by activation of PKA, while inhibition of fibroblast proliferation was mediated exclusively by activation of Epac-1. PGE(2) and Epac-1 inhibited cell proliferation through activation of the small GTPase Rap1, since decreasing Rap1 activity by transfection with Rap1GAP or the dominant-negative Rap1N17 prevented, and transfection with the constitutively active Rap1V12 mimicked, the anti proliferative effects of PGE(2). On the other hand, PKA inhibition of collagen was dependent on inhibition of protein kinase C-delta. The selective use of PKA and Epac-1 pathways to inhibit distinct aspects of fibroblast activation illustrate the pleiotropic ability of PGE(2) to inhibit diverse fibroblast functions. PMID- 18421015 TI - Endogenous beta-adrenergic receptors inhibit lipopolysaccharide-induced pulmonary cytokine release and coagulation. AB - Beta2-adrenergic receptors are expressed on different cell types in the lung, including respiratory epithelial cells, smooth muscle cells, and macrophages. The aim of the current study was to determine the role of beta-adrenergic receptors in the regulation of lung inflammation induced by instillation via the airways of lipopolysaccharide (LPS) (a constituent of the gram-negative bacterial cell wall) or lipoteichoic acid (LTA) (a component of the gram-positive bacterial cell wall). Mice inhaled the beta-adrenergic antagonist propranolol or saline 30 minutes before and 3 hours after intranasal LPS or LTA administration. LPS and LTA induced a profound inflammatory response in the lungs as reflected by an influx of neutrophils and the release of proinflammatory cytokines and chemokines into bronchoalveolar lavage fluid (BALF). Propranolol inhalation resulted in enhanced LPS-induced lung inflammation, which was reflected by a stronger secretion of TNF-alpha, IL-6, and monocyte chemoattractant protein-1 into BALF and by enhanced coagulation activation (thrombin-antithrombin complexes). In LTA induced lung inflammation, propranolol did not influence cytokine release but potentiated activation of coagulation. Propranolol did not alter neutrophil recruitment in either model. This study suggests that beta-adrenergic receptors, which are widely expressed in the lungs, serve as negative regulators of pulmonary cytokine release and coagulation induced by LPS and less so during LTA induced pulmonary inflammation. PMID- 18421014 TI - Curcumin restores corticosteroid function in monocytes exposed to oxidants by maintaining HDAC2. AB - Oxidative stress as a result of cigarette smoking is an important etiologic factor in the pathogenesis of chronic obstructive pulmonary disease (COPD), a chronic steroid-insensitive inflammatory disease of the airways. Histone deacetylase-2 (HDAC2), a critical component of the corticosteroid anti inflammatory action, is impaired in lungs of patients with COPD and correlates with disease severity. We demonstrate here that curcumin (diferuloylmethane), a dietary polyphenol, at nanomolar concentrations specifically restores cigarette smoke extract (CSE)- or oxidative stress-impaired HDAC2 activity and corticosteroid efficacy in vitro with an EC(50) of approximately 30 nM and 200 nM, respectively. CSE caused a reduction in HDAC2 protein expression that was restored by curcumin. This decrease in HDAC2 protein expression was reversed by curcumin even in the presence of cycloheximide, a protein synthesis inhibitor. The proteasomal inhibitor, MG132, also blocked CSE-induced HDAC2 degradation, increasing the levels of ubiquitinated HDAC2. Biochemical and gene chip analysis indicated that curcumin at concentrations up to 1 muM propagates its effect via antioxidant-independent mechanisms associated with the phosphorylation-ubiquitin proteasome pathway. Thus curcumin acts at a post-translational level by maintaining both HDAC2 activity and expression, thereby reversing steroid insensitivity induced by either CSE or oxidative stress in monocytes. Curcumin may therefore have potential to reverse steroid resistance, which is common in patients with COPD and asthma. PMID- 18421016 TI - Cystic adenomatoid malformations are induced by localized FGF10 overexpression in fetal rat lung. AB - Fibroblast growth factor-10 (FGF10) is a mesenchymal growth factor, involved in epithelial and mesenchymal interactions during lung branching morphogenesis. In the present work, FGF10 overexpression was transiently induced in a temporally and spatially restricted manner, during the pseudoglandular or canalicular stages of rat lung development, by trans-uterine ultrasound-guided intraparenchymal microinjections of adenoviral vector encoding the rfgf10 transgene. The morphologic and histologic classification of the resulting malformations were dependent upon developmental stage and location. Overexpression of FGF10 restricted to the proximal tracheobronchial tree during the pseudoglandular phase resulted in large cysts lined by tall columnar epithelium composed primarily of Clara cells with a paucity of Type II pneumocytes, resembling bronchiolar type epithelium. In contrast, FGF10 overexpression in the distal lung parenchyma during the canalicular phase resulted in small cysts lined by cuboidal epithelial cells composed of primarily Type II pneumocytes resembling acinar epithelial differentiation. The cystic malformations induced by FGF10 overexpression appear to closely recapitulate the morphology and histology of the spectrum of human congenital cystic adenomatoid malformation (CCAM). These findings support a role for FGF10 in the induction of human CCAM and provide further mechanistic insight into the role of FGF10 in normal and abnormal lung development. PMID- 18421017 TI - Loss of RAGE in pulmonary fibrosis: molecular relations to functional changes in pulmonary cell types. AB - The receptor for advanced glycation end products (RAGE) is a transmembrane receptor of the Ig superfamily. While vascular RAGE expression is associated with kidney and liver fibrosis, high expression levels of RAGE are found under physiological conditions in the lung. In this study, RAGE expression in idiopathic pulmonary fibrosis was assessed, and the relationship of the receptor to functional changes of epithelial cells and pulmonary fibroblasts in the pathogenesis of the disease was investigated. Significant down-regulation of RAGE was observed in lung homogenate and alveolar epithelial type II cells from patients with idiopathic pulmonary fibrosis, as well as in bleomycin-treated mice, demonstrated by RT-PCR, Western blotting, and immunohistochemistry. In vitro, RAGE down-regulation was provoked by stimulation of primary human lung fibroblasts and A549 epithelial cells with the proinflammatory cytokines, transforming growth factor-beta1 or TNF-alpha. Blockade of RAGE resulted in impaired cell adhesion, and small interfering RNA-induced knockdown of RAGE increased cell proliferation and migration of A549 cells and human primary fibroblast in vitro. These results indicate that RAGE serves a protective role in the lung, and that loss of the receptor is related to functional changes of pulmonary cell types, with the consequences of fibrotic disease. PMID- 18421018 TI - The effects of leptin on airway smooth muscle responses. AB - Obesity is associated with asthma and airway hyperresponsiveness. Leptin modulates some of the proinflammatory effects observed in obesity. The objective of this study was to determine the effects of leptin on airway smooth muscle responses. The effect of leptin (0.1-100 ng/ml) on migration (toward platelet derived growth factor [PDGF], 10 ng/ml, across collagen-coated membrane in Transwell culture plates), proliferation (by BrDU incorporation), and cytokine production (by Bioplex bead assay) of cultured human airway smooth muscle cells from nine nonasthmatic donors was assessed. Effects of leptin on the contractile responses were studied in bovine tracheal smooth muscle rings. Leptin receptor expression and activation of STAT-3, Src kinase, Suppressor of Cytokine Signaling 3 (SOCS-3), and COX were evaluated by Western blotting and PCR. PGE(2) levels in supernatant were assessed by enzyme immunoassay. Human airway smooth muscle cells express leptin receptor, which, when engaged, phosphorylated STAT-3. Leptin inhibited PDGF-induced human airway smooth muscle migration and proliferation and IL-13-induced eotaxin production. Leptin did not stimulate cytokine synthesis and did not evoke contractile responses or inhibit isoproterenol-induced relaxation of carbachol-induced contraction of bovine tracheal rings. The inhibitory effects on migration and eotaxin production are not due to activation of SOCS-3 but are partly due to increased production of PGE(2) because they were attenuated by indomethacin. In conclusion, leptin inhibited human airway smooth muscle proliferation, migration toward PDGF, and IL-13-induced eotaxin production. This is partly mediated by PGE(2) secretion from smooth muscle cells induced by leptin. The association between obesity and asthma is unlikely to be due to a direct effect of leptin on airway smooth muscle. PMID- 18421019 TI - Expression and function of toll-like receptors at the maternal-fetal interface. AB - Toll-like receptors (TLRs) form the major family of pattern recognition receptors that are involved in innate immunity. Innate immune responses against microorganisms at the maternal-fetal interface may have a significant impact on the success of pregnancy because intrauterine infections have been shown to be strongly associated with certain complications of pregnancy. At the maternal fetal interface, TLRs are expressed not only in the immune cells but also in nonimmune cells such as trophoblasts and decidual cells. Moreover, their expression patterns vary according to the stage of pregnancy. Here we will describe potential functions of TLRs in these cells, their recognition and response to microorganisms, and their involvement in the innate immunity. The impact of TLR-mediated innate immune response will be discussed via animal model studies, as well as clinical observations. PMID- 18421021 TI - Shotgun proteomic analysis of vaginal fluid from women in late pregnancy. AB - Liquid chromatography and tandem mass spectrometry without prior fractionation (shotgun proteomics) were used to analyze vaginal fluid from patients admitted for signs and symptoms of preterm labor. The patients had an average age of 26.3 +/- 5.9 years, a gestational age of 30.5 +/- 2.5 weeks, and a median cervical dilation of 1 cm (range, 0-6 cm). None of the patients exhibited signs of vaginal infection at the time of enrollment. Shotgun proteomics yielded reproducible identifications (R = 0.973) of more than 40 proteins in vaginal fluid samples, such as plasma proteins, epithelial structural proteins, and several immunoregulatory proteins, including some that were previously linked to intra amniotic infection. This initial characterization of the vaginal fluid proteome using a nonbiased, high-throughput technique yields reproducible results in late pregnancy. The presence of host defense proteins in vaginal fluid suggests that this technique may be useful for future study of inflammation-related preterm birth. PMID- 18421020 TI - The impact of ACTH receptor knockdown on fetal and adult ovine adrenocortical cell function. AB - Preparing the mammalian fetus for birth requires an increase in fetal plasma glucocorticoid levels. The mechanisms facilitating this increase are not fully known. It has been shown in sheep that the prepartum elevation in fetal plasma cortisol is accompanied by increases in adrenocorticotropin receptor (ACTH-R) expression in the fetal adrenal and in the adrenal responsiveness to stimulation. To determine the significance of the upregulation in ACTH-R expression on fetal adrenal function, the authors used small interfering RNA targeted to the ovine ACTH-R to reduce receptor expression and studied responses to stimulation in ovine adrenal cells. They studied fetal cells from late gestation after responsiveness had increased. They also studied adult cells to determine if maturation would influence the impact of receptor expression suppression on responsiveness. Fetal and adult cells were obtained, dispersed, transfected with receptor-targeted small interfering RNA or scrambled small interfering RNA, and subsequently stimulated with ACTH. Cells and media were harvested for measurements of gene and protein expression and cyclic adenosine monophosphate (cAMP) and cortisol levels. The ability of ACTH to upregulate its receptor or steroid acute regulatory protein was attenuated in fetal (P < .01) and adult cells (P < .01) by small interfering RNA treatment; the blockade was more pronounced in the adult cells (P < .01). The small interfering RNA treatment also blocked the cAMP response to ACTH in fetal (P < .001) and adult (P < .05) cells. This was accompanied by marked reductions in cortisol responses in both (P < .001 and P < .01, respectively). These data suggest that upregulation of the ACTH-R expression in late gestation is essential for the increase in adrenal steroidogenic capacity occurring then. The data also indicate that a reduction in the ACTH-R expression blocks the ability of the peptide to stimulate early steps in the steroidogenic pathway event after maturation is complete. PMID- 18421022 TI - Peripubertal hyperinsulinemia upregulates phosphatidylinositol 3-kinase/akt pathway in rat ovaries. AB - OBJECTIVE: Polycystic ovary syndrome (PCOS) usually develops during peripuberty and is often associated with hyperinsulinemia. Paradoxically, hyperinsulinemic patients with PCOS develop peripheral insulin resistance but retain ovarian insulin sensitivity. We investigated the effect of peripubertal hyperinsulinemia on insulin signaling pathways in rat ovaries. METHODS: Hyperinsulinemia was induced in peripubertal female rats by infusing insulin (0.14 IU/day) for 4 weeks. Control animals received normal saline. At autopsy, trunk blood was collected for insulin assay; ovaries were collected for examining the effect of hyperinsulinemia on phosphatidylinositol 3-kinase/Akt (PI3-K/Akt) and mitogen activated protein kinase (MAPK/ERK1/2) pathways. RESULTS: Compared with control, ovarian protein levels of total Akt, phospho-Akt, and phospho-glycogen synthase kinase-3beta were upregulated and phospho-phosphatase and tensin homolog was downregulated in hyperinsulinemic animals. The MAPK/ERK1/2 pathway was unaffected. CONCLUSIONS: Peripubertal hyperinsulinemia upregulates the PI3-K/Akt pathway in rat ovaries. Ovarian insulin sensitivity in hyperinsulinemic patients with PCOS is potentially retained by this mechanism. PMID- 18421023 TI - Differential gene expression profiling in HELLP syndrome placentas. AB - The aim of this study was to identify differentially expressed genes by suppression subtractive hybridization (SSH) in HELLP placentas. Two cDNA libraries were constructed; HSI (HELLP subtracted induced or upregulated) and HSS (HELLP subtracted suppressed or downregulated). Two hundred eighty-eight cDNA clones were sequenced; 37 were matched to GenBank entries and included genes in cell communication and organization, cellular processes, genetic information processing, and metabolic processes. A subgroup of 11 genes of interest was further selected for real-time quantitative polymerase chain reaction confirmation. Results showed no differences in expression of chosen upregulated genes between HELLP and non-HELLP placentas; 6 HELLP downregulated genes were significantly suppressed. Two genes related to production of secreted proteins, CTHRC1 and SERPINE2. SERPINE2 (PAI-1) is a soluble protease inhibitor and is a potential biomarker by Western blot analysis, and the protein is significantly decreased in HELLP placentas. SERPINE2 might be tested clinically in patients for early diagnosis of HELLP syndrome. PMID- 18421024 TI - Jugular lymphatic maldevelopment in Turner syndrome and trisomy 21: different anomalies leading to nuchal edema. AB - Increased nuchal translucency (NT), morphologically known as nuchal edema, is an ultrasound marker for aneuploidy. Turner syndrome presents with massive NT, called cystic hygroma. Conflicting data exist as to whether cystic hygroma and increased NT are different entities. Both are associated with jugular lymphatic distension. The authors investigated jugular lymphatics of trisomy 21, Turner syndrome, and normal karyotype fetuses. Fetuses were investigated using immunohistochemistry for blood vascular, lymphatic, and smooth muscle cell markers. Trisomy 21 fetuses showed nuchal cavities within the mesenchymal edema negative for endothelial markers. These were extremely large in Turner fetuses, showing similar characteristics. The skin showed numerous dilated lymphatics in the case of trisomy 21 and scanty small lymphatics in Turner fetuses. A jugular lymphatic sac was present in control and trisomy 21 fetuses and was enlarged in trisomy 21 cases. In Turner fetuses, no jugular lymphatic sac was observed. Nuchal edema in trisomy 21 and Turner syndrome appears to be a similar entity caused by different lymphatic abnormalities. PMID- 18421025 TI - A digenic combination of polymorphisms within ESR1 and ESR2 genes are associated with age at menarche in the Spanish population. AB - In the present study, the authors look at an association of genetic variants within estrogen synthesis and signaling pathways and age at menarche (AAM) in Spanish women. They analyzed 9 polymorphisms in 6 different genes in 714 well characterized postmenopausal women from Spain. They performed a quantitative trait locus study of these markers individually or in digenic combinations in relation to AAM. None of the studied markers, with the exception of the follicle stimulating hormone receptor (P = .013), were significantly associated with AAM in the Spanish population, and no marker demonstrated an association of statistical significance after multiple testing corrections (P > .0055). In contrast, linear regression analysis suggests epistatic interactions including ESR1 and ESR2 loci in relation to AAM in the series (P = .003). The results suggest that epistatic interactions of ESR1 and ESR2 alleles could be associated with advancing AAM among Spanish women. PMID- 18421026 TI - Effect of relaxin on TGF-beta1 expression in cultured vaginal fibroblasts from women with stress urinary incontinence. AB - The objective of this study is to compare relaxin's effect on transforming growth factor (TGF)- beta1 and latent TGF-beta1-binding protein (LTBP-1) in vaginal fibroblasts from women with stress urinary incontinence (SUI) to continent women (controls) in both phases of the menstrual cycle. Fibroblasts were treated with relaxin. TGF-beta1 levels were measured by enzyme-linked immunosorbent assay. LTBP-1 expression was evaluated by Western blot. In the proliferative phase, total TGF-beta1 level in the supernatant, cells, and extracellular matrix (ECM) of SUI fibroblasts decreased with increasing relaxin concentration (P < .05). Active TGF-beta1 levels increased at a low concentration of relaxin (P < .05) in the supernatant but decreased in the ECM of SUI fibroblasts at high concentration (P < .05). In the secretory phase, total TGF-beta1 levels decreased with relaxin treatment (P < .05) in the supernatant, cells and ECM of both women with SUI and controls. Relaxin decreased the levels of total and active TGF-beta1 in the ECM isolated from SUI vaginal fibroblasts. PMID- 18421027 TI - Lipoxygenase pathway receptor expression in ovarian cancer. AB - OBJECTIVE: To determine the expression of lipoxygenase (LOX) pathway receptors in ovarian cancer as a potential target for anti-LOX-based therapy. STUDY DESIGN: Paraffin-embedded tumor samples from epithelial ovarian cancer patients were used to construct tissue microarrays to stain for the proposed sites of inhibition of a LOX inhibitor (5-LOX, LTB4-BLT1, and LTB4-BLT2). RESULTS: 245 samples were available for interpretation. Strong expression was demonstrated in 45%, 34%, and 6% of ovarian cancer for LTB4-BLT2, LTB4-BLT1, and 5-LOX, respectively. Expression of LTB4-BLT2 correlated with advanced stage III/IV disease (P = .05), suboptimal debulking (P = .07), and platinum resistance (P = .03). No correlation was seen with regard to disease-free survival. CONCLUSIONS: LOX pathway receptor expression was found in the majority of cancers evaluated. Additionally, LTB4 BLT2 expression portends worse clinical parameters for ovarian cancer. Thus, further investigation on the role of LOX pathway in ovarian cancer is warranted. PMID- 18421029 TI - Markers of inflammation in patients with symptomatic and asymptomatic carotid artery stenosis: a case-control study. AB - OBJECTIVES: There is evidence that inflammation plays a role in the pathogenesis of atherosclerosis. We compared levels of inflammatory markers between patients undergoing carotid endarterectomy (CEA) and controls, and between patients with symptomatic and asymptomatic internal carotid artery (ICA) stenosis. MATERIALS AND METHODS: A total of 180 patients with ICA stenosis were compared with 180 age matched and sex-matched controls. The biomarkers evaluated were high-sensitivity C-reactive protein (hs-CRP), soluble intercellular adhesion molecule 1, soluble vascular cell adhesion molecule 1 (sVCAM-1), and interleukin-6 (IL-6). RESULTS: The levels of hs-CRP, sVCAM-1, and IL-6 in the CEA group were significantly higher than in the control group (1.87 mg/mL vs 1.44 mg/mL, P = .011; 1408 ng/dL vs 672 ng/dL, P < .001; 11.9 pg/mL vs 6.3 pg/mL, P < .001). Multivariate linear regression analysis, adjusted for all clinical and physiologic parameters, showed a significant association between ICA stenosis and hs-CRP, sVCAM-1, and IL-6 concentrations. Analysis of symptomatic (n = 101) and asymptomatic (n = 79) ICA stenosis did not detect a difference in levels of these markers. CONCLUSIONS: Our study suggests that inflammatory markers could serve as markers for ICA atherosclerosis but are not useful to identify carotid plaque at risk for symptomatic conversion. PMID- 18421030 TI - Are statins an option in the management of abdominal aortic aneurysms? AB - Medline was searched for studies investigating the perioperative and long-term results that derive from statin use in patients with abdominal aortic aneurysm and the clinical and experimental evidence dealing with aneurysm expansion. Data suggest that statins improve the perioperative and long-term outcomes of aneurysm operations and may also reduce expansion rates. International guidelines recommend the use of statins in these patients because abdominal aortic aneurysms are considered as a coronary heart disease equivalent. These guidelines do not appear to have been widely implemented. Preliminary results suggest that statins might play a role in the management of abdominal aortic aneurysms. Verification of these results in large-scale trials may hold implications for a more comprehensive approach of patients with abdominal aortic aneurysms as well as population-wide aneurysm screening programs. Irrespective of the role of statins on aneurysm expansion rates, it is important to ensure that all abdominal aortic aneurysm patients receive statin therapy to improve perioperative and long-term morbidity and mortality. PMID- 18421031 TI - Popliteal artery aneurysm surgery: the role of emergency setting. AB - The aim of this study was to compare the outcomes of emergent surgery caused by acute complications versus elective surgery of popliteal artery aneurysms (PAAs) and to evaluate the advantages of elective repair related to limb salvage and bypass patency rates. Fifty PAAs were operated on in 40 patients from January 2000 to December 2004. Surgery was elective in 34 cases (68%) and emergent in 16 (32%). Emergent repair was performed because of acute complications. All patients were treated surgically. Early results in terms of limb salvage, primary patency, and assisted patency were assessed. Follow-up consisted of clinical and/or ultrasonographic examinations at 10 days and 1, 3, 6, and 12 months and yearly thereafter. Long-term limb salvage and bypass patency rates were analyzed. The mean follow-up was 17 months. The 1-year limb salvage rate in the elective group was 97.1% versus 56.3% in the emergent group (P = .0007). The bypass patency rate at 1 year was 94.1% in the elective group versus 66.7% in the emergent group (P = .03). In this study, the outcomes of the PAA repair were significantly better in the group operated electively and without acute symptoms at presentation compared with the group with acute complications, operated on emergently, especially related to limb salvage and bypass patency rates. PMID- 18421032 TI - Endovascular retrieval of an intraaortic greenfield vena cava filter. AB - Aberrant placement of vena cava filters has been documented. Only one case of intraaortic deployment, in which the filter was left at the aortic bifurcation with no adverse effects over a 4-year follow-up period, has been reported. We describe the endovascular retrieval of an intraaortic Greenfield filter using a snare and large sheath to protect the aortic intima from injury during removal of the device. PMID- 18421033 TI - Surgical complications in a hemophilia patient with factor VIII inhibitor and their endovascular management. AB - Hemophilia, an X-linked disorder due to clotting factors VIII or IX deficiency, is associated with recurrent hemorrhage into joints, soft tissue, and organs. Recombinant factor VIII concentrates are available for the treatment of bleeding complications. However, inhibitors to factor VIII develop in 25% to 35% of patients with severe hemophilia A, making the management of bleeding episodes more complex. The treatment of unusual metachronous ruptured aneurysms in a patient with hemophilia A with factor VIII inhibitor is presented, and the outcomes of endovascular and open operations are discussed. PMID- 18421034 TI - Endovascular repair of femoral artery pseudoaneurysm after orthopedic surgery with balloon-expandable covered stents. AB - Arterial injury after orthopedic procedures is an uncommon complication that can present clinically in a variety of forms and has conventionally been repaired by open vascular surgery. The case and discussion in this article highlights the usefulness of endovascular repair following a delayed presentation of vascular injury from an orthopedic procedure. PMID- 18421035 TI - Surgical repair of a gunshot injury to the left carotid artery: case report and review of literature. AB - We present a case of a 37-year-old man who sustained a single gunshot wound (penetrating zones I and II) and internal carotid artery injury. Optimal evaluation and management of vascular injury remains controversial. Literature on the operative techniques of carotid artery injuries is reviewed. PMID- 18421036 TI - Endovascular exclusion of superior mesenteric artery pseudoaneurysms: an alternative to open laparotomy in high-risk patients. AB - Superior mesenteric artery (SMA) aneurysms are rare. However, patients are frequently symptomatic on presentation and require urgent repair. SMA aneurysms have the potential for severe complications, including thrombosis or rupture, resulting in acute mesenteric ischemia and death. In patients with adhesions secondary to prior abdominal surgery, traditional open exposure of the aneurysm neck via dissection at the base of the transverse colon mesentery may be technically difficult. Endovascular exclusion of visceral artery aneurysms using covered stent grafts presents a reasonable alternative to the morbidity associated with laparotomy. We report an interesting case of a patient with a large SMA aneurysm and multiple prior laparotomies treated percutaneously with a combined expanded polytetrafluoroethylene/nitinol self-expanding stent-graft allowing maintenance of end-organ perfusion and bowel viability. PMID- 18421037 TI - Prolonged success with a covered endovascular stent after emergent use in radiation-induced subclavian artery blowout: a case report. AB - Radiation-induced damage to the bone, soft tissues, and vasculature represents the unfortunate consequences of radiation therapy for the treatment of malignant tumors. Complications arising from irradiation are frequently challenging to manage and may be life threatening. A case is presented of a patient with a longstanding clavicular osteoradionecrosis with an acute massive hemorrhage after rupture of the subclavian artery and subsequent management with endovascular stent placement. With over 2 years' follow-up, vascular patency was maintained with no further bleeding episodes in this surgically high-risk patient. PMID- 18421038 TI - Interposition grafting of popliteal artery cystic adventitial disease: case report. AB - Cystic adventitial disease of the popliteal artery is a rare cause of leg claudication occurring primarily in young adults. We report a case of a 41-year old athletic man who presented with rapidly progressive left leg claudication. Using duplex ultrasound and magnetic resonance angiography, a diagnosis of popliteal artery cystic adventitial disease was made. The cystic popliteal artery was resected and replaced using a prosthetic interposition graft. At the 1.5-year follow-up, the cysts have recurred; however, the related symptoms have not. This is in contrast to a previously reported case of interposition saphenous vein grafting requiring resection due to invasion by recurrent cysts within 6 months. PMID- 18421039 TI - Platelet activation in bypass surgery for critical limb ischemia. PMID- 18421041 TI - What constitutes "improved prognosis"? PMID- 18421042 TI - The multi-institutional phase I study: disadvantages without advantages? PMID- 18421043 TI - All dex'ed out with nowhere to go? PMID- 18421044 TI - Assessing the measure of a new drug: is survival the only thing that matters? PMID- 18421045 TI - From silos to a critical path of new agent development: a paradigm to revolutionize clinical research. PMID- 18421046 TI - Multi-institutional phase I trials of anticancer agents. AB - PURPOSE: Physicians involved in the conduct of phase I studies of novel anticancer agents have raised concerns about the emergence of multi-institutional phase I trials and about using the optimal biologic dose (OBD) as an alternative to the maximum-tolerated dose (MTD) as the primary end point in early drug development. We sought to determine the factors associated with multi institutional phase I studies and OBD determination. PATIENTS AND METHODS: We reviewed all published phase I trials between January 1998 and June 2006 from two major clinical cancer journals. The following components from each trial were determined: number of participating sites, sponsor, nation where study was conducted, MTD or OBD established, number of patients accrued, mechanism of action of the studied agent, accrual time, and tumor type. RESULTS: We identified 463 trials. Fifty-six percent were performed in single institutions. Only 30% reported accrual time. The number of patients enrolled on single institution studies was significantly lower than on multi-institutional studies (P < .05), but there was no difference in accrual time. There was no association between the number of institutions and the sponsor or the mechanism of drug action. National Institutes of Health-sponsored trials enrolled fewer patients per trial than pharmaceutical-sponsored trials (P < .05). Although 99% of trials with cytotoxic agents determined an MTD, only 64% of trials with targeted agents did. CONCLUSION: Multi-institutional phase I studies do not decrease the time to study completion and result in an increase in number of patients per trial. One third of trials with targeted agents failed to determine an MTD. PMID- 18421047 TI - Asparaginase may influence dexamethasone pharmacokinetics in acute lymphoblastic leukemia. AB - PURPOSE: Dexamethasone is used widely in oncology, but pharmacokinetic studies are lacking. We evaluated dexamethasone pharmacokinetics in children with acute lymphoblastic leukemia. PATIENTS AND METHODS: We assessed 214 children with acute lymphoblastic leukemia who received 418 courses of oral dexamethasone (8 mg/m(2)/d) on days 1 and 8 of reinduction. Extensive asparaginase use preceded reinduction in the 101 children in the standard/high-risk treatment arm but not in the 113 children in the low-risk treatment arm. A one-compartment model with first-order absorption and disposition was fit to dexamethasone plasma concentrations by using maximum a posteriori probability estimation; we evaluated covariates by using linear mixed models. RESULTS: Interpatient and intrapatient variabilities in apparent clearance were substantial; they were 46% and 53%, respectively. Variability was explained by the serum albumin concentration (P < .0001), concomitant use of fentanyl (P = .008) and ketoconazole (P = .03), and age (P = .006). Apparent clearance was higher in the low-risk arm (P < .001) and was related to a greater serum albumin concentration (P < .001) and to a lower exposure to asparaginase than in the standard/high-risk arm. Hypoalbuminemia, a biomarker of asparaginase activity, was associated with a lower dexamethasone apparent clearance (P = .04) in patients in the standard/high-risk arm that was more pronounced in those not allergic to asparaginase. Ethnicity or gender did not explain apparent clearance variability. CONCLUSION: Dexamethasone pharmacokinetics are highly variable and are related to the concurrent use of particular drugs, age, and treatment intensity. Patients allergic to asparaginase may be doubly disadvantaged: they not only suffer from diminished exposure to asparaginase but also, by maintaining high clearance of dexamethasone, may experience fewer antileukemic effects of dexamethasone. PMID- 18421048 TI - Phase I study of MGCD0103 given as a three-times-per-week oral dose in patients with advanced solid tumors. AB - PURPOSE: MGCD0103 is a novel isotype-selective inhibitor of human histone deaceylases (HDACs) with the potential to regulate aberrant gene expression and restore normal growth control in malignancies. PATIENTS AND METHODS: A phase I trial of MGCD0103, given as a three-times-per-week oral dose for 2 of every 3 weeks, was performed in patients with advanced solid tumors. Primary end points were safety, tolerability, pharmacokinetics (PK), pharmacodynamic (PD) assessments of HDAC activity, and histone acetylation status in peripheral WBCs. RESULTS: Six dose levels ranging from 12.5 to 56 mg/m(2)/d were evaluated in 38 patients over 99 cycles (median, 2; range, 1 to 11). The recommended phase II dose was 45 mg/m(2)/d. Dose-limiting toxicities consisting of fatigue, nausea, vomiting, anorexia, and dehydration were observed in three (27%) of 11 and two (67%) of three patients treated at the 45 and 56 mg/m(2)/d dose levels, respectively. Disease stabilization for four or more cycles was observed in five (16%) of 32 patients assessable for efficacy. PK analyses demonstrated interpatient variability which was improved by coadministration with low pH beverages. Elimination half-life ranged from 6.7 to 12.2 hours, and no accumulation was observed with repeated dosing. PD evaluations confirmed inhibition of HDAC activity and induction of acetylation of H3 histones in peripheral WBCs from patients by MGCD0103. CONCLUSION: At doses evaluated, MGCD0103 appears tolerable and exhibits favorable PK and PD profiles with evidence of target inhibition in surrogate tissues. PMID- 18421049 TI - Taxanes alone or in combination with anthracyclines as first-line therapy of patients with metastatic breast cancer. AB - PURPOSE: Taxanes (paclitaxel or docetaxel) have been sequenced or combined with anthracyclines (doxorubicin or epirubicin) for the first-line treatment of advanced breast cancer. This meta-analysis uses data from all relevant trials to detect any advantages of taxanes in terms of tumor response, progression-free survival (PFS), and survival. PATIENTS AND METHODS: Individual patient data were collected on eight randomized combination trials comparing anthracyclines + taxanes (+ cyclophosphamide in one trial) with anthracyclines + cyclophosphamide (+ fluorouracil in four trials), and on three single-agent trials comparing taxanes with anthracyclines. Combination trials included 3,034 patients; single agent trials included 919 patients. RESULTS: Median follow-up of living patients was 43 months, median survival was 19.3 months, and median PFS was 7.1 months. In single-agent trials, response rates were similar in the taxanes (38%) and in the anthracyclines (33%) arms (P = .08). The hazard ratios for taxanes compared with anthracyclines were 1.19 (95% CI, 1.04 to 1.36; P = .011) for PFS and 1.01 (95% CI, 0.88 to 1.16; P = .90) for survival. In combination trials, response rates were 57% (10% complete) in taxane-based combinations and 46% (6% complete) in control arms (P < .001). The hazard ratios for taxane-based combinations compared with control arms were 0.92 (95% CI, 0.85 to 0.99; P = .031) for PFS and 0.95 (95% CI, 0.88 to 1.03; P = .24) for survival. CONCLUSION: Taxanes were significantly worse than single-agent anthracyclines in terms of PFS, but not in terms of response rates or survival. Taxane-based combinations were significantly better than anthracycline-based combinations in terms of response rates and PFS, but not in terms of survival. PMID- 18421050 TI - Evaluation of tumor response, disease control, progression-free survival, and time to progression as potential surrogate end points in metastatic breast cancer. AB - PURPOSE: Overall survival (OS) can be observed only after prolonged follow-up, and any potential effect of first-line therapies on OS may be confounded by the effects of subsequent therapy. We investigated whether tumor response, disease control, progression-free survival (PFS), or time to progression (TTP) could be considered a valid surrogate for OS to assess the benefits of first-line therapies for patients with metastatic breast cancer. PATIENTS AND METHODS: Individual patient data were collected on 3,953 patients in 11 randomized trials that compared an anthracycline (alone or in combination) with a taxane (alone or in combination with an anthracycline). Surrogacy was assessed through the correlation between the end points as well as through the correlation between the treatment effects on the end points. RESULTS: Tumor response (survival odds ratio [OR], 6.2; 95% CI, 5.3 to 7.0) and disease control (survival OR, 5.5; 95% CI, 4.8 to 6.3) were strongly associated with OS. PFS (rank correlation coefficient, 0.688; 95% CI, 0.686 to 0.690) and TTP (rank correlation coefficient, 0.682; 95% CI, 0.680 to 0.684) were moderately associated with OS. Response log ORs were strongly correlated with PFS log hazard ratios (linear coefficient [rho], 0.96; 95% CI, 0.73 to 1.19). Response and disease control log ORs and PFS and TTP log hazard ratios were poorly correlated with log hazard ratios for OS, but the confidence limits of rho were too wide to be informative. CONCLUSION: No end point could be demonstrated as a good surrogate for OS in these trials. Tumor response may be an acceptable surrogate for PFS. PMID- 18421051 TI - Phase II trial of lapatinib for brain metastases in patients with human epidermal growth factor receptor 2-positive breast cancer. AB - PURPOSE: One third of women with advanced human epidermal growth factor receptor 2 (HER-2)-positive breast cancer develop brain metastases; a subset progress in the CNS despite standard approaches. Medical therapies for refractory brain metastases are neither well-studied nor established. We evaluated the safety and efficacy of lapatinib, an oral inhibitor of epidermal growth factor receptor (EGFR) and HER-2, in patients with HER-2-positive brain metastases. PATIENTS AND METHODS: Patients had HER-2-positive breast cancer, progressive brain metastases, prior trastuzumab treatment, and at least one measurable metastatic brain lesion. Patients received lapatinib 750 mg orally twice a day. Tumor response was assessed by magnetic resonance imaging every 8 weeks. The primary end point was objective response (complete response [CR] plus partial response [PR]) in the CNS by Response Evaluation Criteria in Solid Tumors (RECIST). Secondary end points included objective response in non-CNS sites, time to progression, overall survival, and toxicity. RESULTS: Thirty-nine patients were enrolled. All patients had developed brain metastases while receiving trastuzumab; 37 had progressed after prior radiation. One patient achieved a PR in the brain by RECIST (objective response rate 2.6%, 95% conditional CI, 0.21% to 26%). Seven patients (18%) were progression free in both CNS and non-CNS sites at 16 weeks. Exploratory analyses identified additional patients with some degree of volumetric reduction in brain tumor burden. The most common adverse events (AEs) were diarrhea (grade 3, 21%) and fatigue (grade 3, 15%). CONCLUSION: The study did not meet the predefined criteria for antitumor activity in highly refractory patients with HER-2-positive brain metastases. Because of the volumetric changes observed in our exploratory analysis, further studies are underway utilizing volumetric changes as a primary end point. PMID- 18421052 TI - Systematic survey of therapeutic trials for metastatic colorectal cancer: room for improvement in the critical pathway. AB - PURPOSE: The current strategy of drug development has been criticized as being highly inefficient. In 2004, the US Food and Drug Administration (FDA) released recommendations to improve this process, including a push for increased use of enrichment trials. It is unclear to what extent aspects of this "Critical Path Initiative" have been adopted in trial designs in metastatic colorectal cancer. METHODS: A systematic review was conducted of actively enrolling treatment trials in metastatic colorectal cancer. Trials were identified from the National Cancer Institute's clinicaltrials.gov and Investigative Drug Branch databases. Trials were categorized based on the number of prior treatments allowed, phase of the trial, agent mechanism of action, and FDA approval status of agents under investigation. RESULTS: One hundred two trials are enrolling, with a combined enrollment goal of more than 20,000 patients. Thirteen percent of trials investigated an agent not yet FDA-approved for any oncology indication. The most common study design was a phase II study limited to previously untreated patients; compared with the remaining trials, these phase II trials were more than 10 times more likely to only use agents FDA-approved for colorectal cancer. Three percent of patients were enrolled onto trials enriched for tumor characteristics that were hypothesized to improve clinical benefit. CONCLUSION: Current clinical trials for metastatic colorectal cancer are deficient in the investigation of agents directed at a novel therapeutic target, overuse phase II studies of FDA-approved agents, and fail to incorporate enrichment trial designs as encouraged by the FDA initiative. PMID- 18421053 TI - Randomized phase III study of capecitabine plus oxaliplatin compared with fluorouracil/folinic acid plus oxaliplatin as first-line therapy for metastatic colorectal cancer. AB - PURPOSE: To evaluate whether capecitabine plus oxaliplatin (XELOX) is noninferior to fluorouracil. folinic acid, and oxaliplatin (FOLFOX-4) as first-line therapy in metastatic colorectal cancer (MCRC). PATIENTS AND METHODS: The initial design of this trial was a randomized, two-arm, noninferiority, phase III comparison of XELOX versus FOLFOX-4. After patient accrual had begun, the trial design was amended in 2003 after bevacizumab phase III data became available. The resulting 2 x 2 factorial design randomly assigned patients to XELOX versus FOLFOX-4, and then to also receive either bevacizumab or placebo. We report here the results of the analysis of the XELOX versus FOLFOX-4 arms. The analysis of bevacizumab versus placebo with oxaliplatin-based chemotherapy is reported separately. The prespecified primary end point for the noninferiority analysis was progression free survival. RESULTS: The intent-to-treat population comprised 634 patients from the original two-arm portion of the study, plus an additional 1,400 patients after the start of the amended 2 x 2 design, for a total of 2,034 patients. The median PFS was 8.0 months in the pooled XELOX-containing arms versus 8.5 months in the FOLFOX-4-containing arms (hazard ratio [HR], 1.04; 97.5% CI, 0.93 to 1.16). The median overall survival was 19.8 months with XELOX versus 19.6 months with FOLFOX-4 (HR, 0.99; 97.5% CI, 0.88 to 1.12). FOLFOX-4 was associated with more grade 3/4 neutropenia/granulocytopenia and febrile neutropenia than XELOX, and XELOX with more grade 3 diarrhea and grade 3 hand-foot syndrome than FOLFOX 4. CONCLUSION: XELOX is noninferior to FOLFOX-4 as a first-line treatment for MCRC, and may be considered as a routine treatment option for appropriate patients. PMID- 18421054 TI - Bevacizumab in combination with oxaliplatin-based chemotherapy as first-line therapy in metastatic colorectal cancer: a randomized phase III study. AB - PURPOSE: To evaluate the efficacy and safety of bevacizumab when added to first line oxaliplatin-based chemotherapy (either capecitabine plus oxaliplatin [XELOX] or fluorouracil/folinic acid plus oxaliplatin [FOLFOX-4]) in patients with metastatic colorectal cancer (MCRC). PATIENTS AND METHODS: Patients with MCRC were randomly assigned, in a 2 x 2 factorial design, to XELOX versus FOLFOX-4, and then to bevacizumab versus placebo. The primary end point was progression free survival (PFS). RESULTS: A total of 1,401 patients were randomly assigned in this 2 x 2 analysis. Median progression-free survival (PFS) was 9.4 months in the bevacizumab group and 8.0 months in the placebo group (hazard ratio [HR], 0.83; 97.5% CI, 0.72 to 0.95; P = .0023). Median overall survival was 21.3 months in the bevacizumab group and 19.9 months in the placebo group (HR, 0.89; 97.5% CI, 0.76 to 1.03; P = .077). Response rates were similar in both arms. Analysis of treatment withdrawals showed that, despite protocol allowance of treatment continuation until disease progression, only 29% and 47% of bevacizumab and placebo recipients, respectively, were treated until progression. The toxicity profile of bevacizumab was consistent with that documented in previous trials. CONCLUSION: The addition of bevacizumab to oxaliplatin-based chemotherapy significantly improved PFS in this first-line trial in patients with MCRC. Overall survival differences did not reach statistical significance, and response rate was not improved by the addition of bevacizumab. Treatment continuation until disease progression may be necessary in order to optimize the contribution of bevacizumab to therapy. PMID- 18421055 TI - Validation of patient's self-reported social functioning as an independent prognostic factor for survival in metastatic colorectal cancer patients: results of an international study by the Chronotherapy Group of the European Organisation for Research and Treatment of Cancer. AB - PURPOSE: A recent study identified a prognostic model for survival in metastatic colorectal cancer patients which included WBC count, alkaline phosphatase (AP), number of metastatic sites, and patients' self-reported social functioning. The aim of this research is to validate this model on data from an independent sample. PATIENTS AND METHODS: This validation study is based on a prospective randomized controlled trial in patients with metastatic colorectal cancer conducted by the European Organisation for Research and Treatment of Cancer (EORTC) Chronotherapy Group. Overall, 564 patients in 10 countries were enrolled. For the purpose of this independent validation, patients with health-related quality of life (HRQOL) baseline data were analyzed. HRQOL was assessed using the EORTC Quality of Life Questionnaire C30 (QLQ-C30). The Cox proportional hazards regression model was used for both univariate and multivariate analyses of survival. RESULTS: The previous model with an additional adjustment, by stratification for sex, was replicated and its parameters were confirmed to independently predict survival: WBC count with an hazard ratio (HR) of 1.31 (95% CI, 1.021 to 1.698; P = .034); AP with an HR of 1.53 (95% CI, 1.188 to 1.979; P = .001); number of sites involved with an HR of 1.90 (95% CI, 1.531 to 2.364; P < .0001); and patients' self-reported social functioning with an HR of 0.94 (95% CI, 0.905 to 0.976; P = .001). The latter translates into a 6% increase in the likelihood of an earlier death for every 10-point decrease in the social functioning scale of the EORTC QLQ-C30. CONCLUSION: This study provides confirmatory evidence of the independent prognostic value of patients' self reported social functioning in patients with advanced colorectal cancer. PMID- 18421056 TI - Cognitive and academic consequences of stem-cell transplantation in children. AB - PURPOSE: To describe cognitive and academic outcomes in survivors after pediatric stem-cell transplantation (SCT) through 5-years post-SCT. PATIENTS AND METHODS: A battery of neurocognitive measures were administered before admission and at 1, 3, and 5 years post-SCT for 268 patients who underwent SCT; the study sample is comprised of 158 patients who survived and were evaluated at 1-year post-SCT. Random coefficient models were generated to depict change over time, and to test differences in slope and intercept for medical and demographic predictor variables. RESULTS: In the cohort as a whole, no significant changes were seen in global intelligence quotient and academic achievement. Despite the overall stability, some significant differences in slopes were found based on diagnosis, type of transplantation, use of total-body irradiation (TBI), and presence of graft-versus-host disease (GVHD). However, these differences were small, and of limited clinical significance. In comparison, differences as a function of socioeconomic status (SES) were much larger. SES was a significant determinant of all cognitive and academic outcomes, and the effect size generally dwarfed that of other significant predictor variables. Age, which had previously been identified as an important determinant of outcome, was not significantly predictive of outcome in this cohort. CONCLUSION: The procedure of SCT entails minimal risk of late cognitive and academic sequelae. Subgroups of patients are at relatively higher risk: patients undergoing unrelated donor transplantation, receiving TBI, and those who experience GVHD. However, these differences are small relative to differences in premorbid functioning, particularly those associated with SES. PMID- 18421057 TI - Eosinophilia with FIP1L1-PDGFRA fusion in a patient with chronic myelomonocytic leukemia. PMID- 18421058 TI - Intracranial collision metastases of small-cell lung cancer and malignant melanoma. PMID- 18421059 TI - Major response to imatinib mesylate in KIT-mutated melanoma. PMID- 18421060 TI - Trastuzumab-related cardiotoxicity in the herceptin adjuvant trial. PMID- 18421061 TI - Targeting pelvic lymph nodes in men with intermediate- and high-risk prostate cancer despite two negative randomized trials. PMID- 18421062 TI - Reaction to American Society of Clinical Oncology 2007 update of recommendations for the use of tumor markers in breast cancer. PMID- 18421063 TI - Inconsistent criteria used in American Society of Clinical Oncology 2007 update of recommendations for the use of tumor markers in breast cancer. PMID- 18421064 TI - Temporal changes in quality of life (QoL) of patients with primary brain tumors: raw QoL data from a randomized trial as reference for future studies. PMID- 18421065 TI - Reporting of subset analysis: where do we draw the line? PMID- 18421066 TI - Improved survival with addition of bevacizumab to paclitaxel and carboplatin in elderly patients with non-small-cell lung cancer. PMID- 18421068 TI - The effect of 5 alpha-reductase inhibitors on erectile function. AB - The 5 alpha-reductase inhibitors, which inhibit conversion of testosterone to dihydrotestosterone, are used for miscellaneous clinical applications, including the treatment of benign prostatic hyperplasia and male pattern hair loss, and for possible reduction of the risk of prostate cancer. Erectile dysfunction has been associated with 5 alpha-reductase inhibitors. Overall, reports in the literature suggest rates of erectile dysfunction to be between 0.8%-33% in men using these medications. However, randomized controlled studies report the rates of erectile dysfunction to be between 0.8%-15.8%. The possible risk association is that these medications impact androgen function, which is understood to contribute to normal erectile physiology. The 5 alpha-reductase inhibitors result in a drop in median serum dihydrotestosterone levels by 60%-93% within 2 years, but there is no major change in testosterone levels. In this review, we surveyed studies on erectile dysfunction in patients treated with 5 alpha-reductase inhibitors and critically examined the evidence that associates 5 alpha-reductase inhibitors and erectile dysfunction. We conclude that 5 alpha-reductase inhibitors do not lead to erectile dysfunction to a significant degree, and we support the position that dihydrotestosterone is less relevant than testosterone in erectile function. PMID- 18421069 TI - Leptin, ghrelin, and adiponectin evaluation in transsexual subjects during hormonal treatments. AB - Gender differences in leptin, ghrelin, and adiponectin levels have been described in a normal population. This is important for understanding differences between males and females in the regulation of food intake, weight gain, body fat distribution, and cardiovascular risk. It is unclear how endogenous and exogenous sex hormones may regulate circulating levels of these factors. Transsexuals during hormonal treatment may represent an ideal model to ascertain the role of exogenous sex hormones on these parameters. In this study, our objective was to evaluate adiponectin, ghrelin, and leptin levels in transsexual subjects during hormone therapy and to compare the results of males and females. Subjects were 26 nondiabetic transsexuals, which included 15 male-to-female (M-to-F, group 3) and 11 female-to-male (F-to-M, group 4) individuals, and 29 age- and BMI-matched controls, which included 15 males (group 1) and 14 females (group 2). Results showed that leptin levels were significantly lower in group 1 compared with group 2 (P = .04) and group 3 (P = .01); no differences were recorded between the other groups. Adiponectin levels were significantly higher in group 3 compared with group 4 (P = .03). No differences were found between the 4 groups for ghrelin levels. In conclusion, our data confirm the sexual dimorphism in serum leptin levels in normal subjects and demonstrate an increase in M-to-F transsexuals. While ghrelin does not show any sexual differences and seems not to be influenced by exogenous sex hormone administration, the lower adiponectin levels in F-to-M transsexuals during treatment confirm that androgens may decrease plasma adiponectin levels. This latter observation suggests that F-to-M transsexual patients could have a higher cardiovascular risk. PMID- 18421070 TI - Effect of caffeine on erectile function via up-regulating cavernous cyclic guanosine monophosphate in diabetic rats. AB - Erectile dysfunction (ED) is a common complication of diabetes mellitus. Phosphodiesterase-5 (PDE5) inhibitors, which inhibit the breakdown of intracellular cyclic guanosine monophosphate (cGMP), are used to treat diabetic ED. Caffeine, a nonselective PDE inhibitor used in our daily diet, is controversial regarding its effect on erectile function. To investigate the effect of caffeine on erectile function in diabetic rat models and explore the mechanism, male Sprague-Dawley rats were injected with streptozotocin to induce diabetes mellitus. The rats with blood glucose levels above 300 mg/dL were selected for the study. The rats were divided into 4 groups: group A (normal control rats), group B (diabetic rats treated with normal saline), group C (diabetic rats treated with caffeine, 10 mg/kg per day), and group D (diabetic rats treated with caffeine, 20 mg/kg per day). After 8 weeks of treatment, intracavernous pressure (ICP) was measured to assess erectile function. The radioimmunoassay was used to evaluate the level of cGMP in the cavernosum. The ICP and the cavernous cGMP decreased significantly in the diabetic rats compared with normal controls. An 8-week administration of caffeine at the given dosages increased the ICP and cavernous cGMP in diabetic rats. Caffeine consumption improved the erectile function of diabetic rats by up-regulating cavernous cGMP. PMID- 18421071 TI - Metabolism of apical versus basolateral sn-2-monoacylglycerol and fatty acids in rodent small intestine. AB - The metabolic fates of radiolabeled sn-2-monoacylglycerol (MG) and oleate (FA) in rat and mouse intestine, added in vivo to the apical (AP) surface in bile salt micelles, or to the basolateral (BL) surface via albumin-bound solution, were examined. Mucosal lipid products were quantified, and the results demonstrate a dramatic difference in the esterification patterns for both MG and FA, depending upon their site of entry into the enterocyte. For both lipids, the ratio of triacylglycerol to phospholipid (TG:PL) formed was approximately 10-fold higher for delivery at the AP relative to the BL surface. Further, a 3-fold higher level of FA oxidation was found for BL compared with AP substrate delivery. Incorporation of FA into individual PL species was also significantly different, with >2-fold greater incorporation into phosphatidylethanolamine (PE) and a 3 fold decrease in the phosphatidylcholine:PE ratio for AP- compared with BL-added lipid. Overnight fasting increased the TG:PL incorporation ratio for both AP and BL lipid addition, suggesting that metabolic compartmentation is a physiologically regulated phenomenon. These results support the existence of separate pools of TG and glycerolipid intermediates in the intestinal epithelial cell, and underscore the importance of substrate trafficking in the regulation of enterocyte lipid metabolism. PMID- 18421072 TI - Plasma fatty acid binding protein 4 is associated with atherogenic dyslipidemia in diabetes. AB - The aim of this study was to evaluate the impact of adipocyte fatty acid binding protein 4 (FABP4) on the lipid profile in type 2 diabetic subjects. Plasma levels of FABP4 and adiponectin and an extensive lipid profile were analyzed in 169 type 2 diabetic subjects and 105 controls. Type 2 diabetic subjects were categorized according the presence of atherogenic dyslipidemia. Univariate statistical analyses, partial correlation tests, and binary logistic regression models were applied. In type 2 diabetic subjects, FABP4 was positively correlated with plasma triglycerides (P = 0.007), apolipoprotein C-III (apoC-III) (P = 0.009), and all the components of triglyceride-rich lipoproteins, including VLDL triglycerides (P = 0.002), VLDL-cholesterol (P = 0.001), and VLDL apoB (P = 0.001). FABP4 was inversely correlated with apoA-I (P = 0.038), HDL-cholesterol (P = 0.002), and HDL apoA-I (P = 0.010) in type 2 diabetic subjects. These correlations are not significantly affected by age, gender, body mass index, adiponectin, insulin, or any pharmacological treatment. The associations are even stronger when the FABP4/adiponectin ratio is considered. None of these associations were observed in controls. High FABP4 and low adiponectin levels are independent predictors of atherogenic dyslipidemia. In conclusion, FABP4 plasma concentrations hold strong potential for development as a clinical biomarker for atherogenic dyslipidemia, independent of obesity and insulin resistance, in type 2 diabetic subjects. PMID- 18421073 TI - The anti-inflammatory effects of Curcuma longa and Berberis aristata in endotoxin induced uveitis in rabbits. AB - PURPOSE: To investigate the anti-inflammatory effect of topical application of Curcuma longa (C. longa) and Berberis aristata (B. aristata) aqueous extracts on experimental uveitis in the rabbit. METHODS: Anterior uveitis was induced in rabbits by intravitreal injection of lipopolysaccharide from Escherichia coli after pretreatment with C. longa and B. aristata aqueous extracts. Subsequently, the anti-inflammatory activity of C. longa and B. aristata was evaluated by grading the clinical signs and histopathologic changes and estimating the inflammatory cell count, protein, and TNF-alpha levels in the aqueous humor. RESULTS: The anterior segment inflammation in the control group was significantly higher than in both the extract-treated groups, as observed by clinical and histopathologic grading. The inflammatory cell count in the control group was 30.75 +/- 7.33 x 10(5) cells/mL, whereas it was 2.39 +/- 0.59 x 10(5) (P < 0.001 vs. control) and 11.56 +/- 2.44 x 10(5) (P = 0.001 vs. control) cells/mL in the C. longa- and B. aristata-treated groups, respectively. The protein content of the aqueous humor was 18.14 +/- 4.98, 3.16 +/- 0.55 (P < 0.001 vs. control), and 8.24 +/- 1.42 (P < 0.01 vs. control) mg/mL in the control, C. longa-, and B. aristata-treated groups, respectively. The aqueous TNF-alpha level in the control group was 976.29 +/- 66.38 pg/mL and was 311.96 +/- 28.50 (P < 0.0001 vs. control) and 654.09 +/- 47.66 (P < 0.001vs. control) pg/mL in the C. longa- and B. aristata-treated groups, respectively. CONCLUSIONS: Topical instillation of aqueous extracts of C. longa and B. aristata showed potent anti-inflammatory activity against endotoxin-induced uveitis in rabbits. PMID- 18421075 TI - Epithelial thickness changes from the induction of myopia with CRTH RGP contact lenses. AB - PURPOSE: To investigate changes in epithelial thickness after overnight wear of CRTH rigid gas-permeable (RGP) lenses (Paragon Vision Sciences, Mesa, AZ) for the correction of hyperopia. METHODS: Twenty subjects wore a +3.50 D hyperopia correcting CRTH lens on one eye for a single night in an attempt to induce myopia (first study). The untreated eye served as the control. Corneal and epithelial thickness was measured at nine points across the horizontal meridian by OCT. Measurements were obtained the night before lens wear, immediately after lens removal the next morning, and 1, 3, 6, and 12 hours after removal. Measurements were obtained 28 hours later, to observe recovery. Then, the attempted hyperopic corrections of +1.50 and +3.50 D were evaluated, using CRTH lenses in both eyes of 20 subjects for a single night (second study). RESULTS: All values were compared to baseline unless otherwise stated. In the first study, the treated eye's central and midperipheral epithelial thickness increased by 21.5% +/- 8.6% and 13.3% +/- 7.6%, respectively, after lens removal (P < 0.001). The control eye's central epithelial thickness (CET) increased by 7.1% +/- 6.0% (P < 0.05). In the second study, CET increased by 17.6% +/- 8.5% (P < 0.001) in the +3.50 D treated eye and by 13.3% +/- 4.8% (P < 0.001) in the +1.50 D-treated eye. Midperipheral epithelial thickening was 5.9% +/- 4.7% (P < 0.05) in the +3.50 D treated eye and 6.0% +/- 6.3% (P < 0.05) in the +1.50 D-treated eye. CONCLUSIONS: CRTH lenses, designed to correct hyperopia, when worn overnight, caused an increase in CET. The amount of epithelial change seemed to differ with modified lens design. PMID- 18421074 TI - Lack of association between LOXL1 variants and primary open-angle glaucoma in three different populations. AB - PURPOSE: Significant association has recently been reported between pseudoexfoliation glaucoma (XFG) and two single-nucleotide polymorphisms (SNPs), rs3825942, and rs1048661, in the lysyl oxidase-like 1 gene (LOXL1). The purpose of this study was to investigate whether XFG-associated variants of LOXL1 play a significant role in primary open-angle glaucoma in the Caucasian, African American, and Ghanaian (West-African) populations. METHODS: POAG was defined as the presence of glaucomatous optic nerve damage, associated visual field loss, and elevated intraocular pressure (>22 mm Hg in both eyes). Thirteen tagging SNPs were genotyped by allelic discrimination assays in the Caucasian (279 cases and 227 controls), African-American (193 cases and 97 controls), and Ghanaian (170 cases and 138 controls) populations. Allele and genotype frequencies were compared between the cases and controls from each population. RESULTS: None of the SNPs associated with XFG in LOXL1 were significantly associated with POAG in these populations. The risk allele frequencies for rs2165241 and rs3825942 were significantly lower in the African-American and Ghanaian populations, compared with Caucasian individuals. CONCLUSIONS: There was no association between SNPs in the LOXL1 gene and POAG. This is the first analysis of the LOXL1 gene in African American and West-African populations. LOXL1 gene variants do not appear to play a significant role in the pathogenesis of POAG in populations of either Caucasian or West-African ancestry. PMID- 18421076 TI - A genome-wide scan maps a novel high myopia locus to 5p15. AB - PURPOSE: This study was conducted to investigate the genetic component of three Chinese pedigrees originating from Hong Kong with autosomal dominant high myopia. METHODS: A whole-genome scan was performed by using microsatellite markers spanning the whole genome with an average spacing of 10 cM. Regions containing markers that yielded LOD scores >1.0 were further analyzed by fine mapping with additional microsatellite markers. Fine-scale mapping of the linkage region was performed by genotyping a set of gene-based SNP markers on a cohort of 94 high myopia cases and 94 control subjects. RESULTS: Two-point LOD scores >1 were observed at markers D5S630, D5S416, D7S510, D11S908, and D17S944. Additional microsatellite markers flanking D5S630 revealed a maximum two-point LOD score of 4.81 at D5S2505 at theta = 0.00. Haplotype analysis narrowed the linkage region to 5p15.33-p15.2 with a 17.45-cM interval. The coding sequences of five genes located within this region, IRX2, IRX1, POLS, CCT5, and CTNND2, were screened. No segregation of polymorphism with high myopia was found. Genotyping of 41 SNPs within this region in a Chinese cohort of 94 high myopia cases and 94 control subjects showed that the allele and genotype distributions of one SNP, rs370010, was different between cases and controls (genotype P = 0.01176, allele P = 0.00271 and trend P = 0.00375), but such association did not remain significant after false discovery rate (FDR) correction. This SNP is located within a hypothetical gene LOC442129. CONCLUSIONS: A novel autosomal dominant high myopia locus was mapped on chromosome 5p15.33-p15.2 with an interval of 17.45 cM. PMID- 18421077 TI - Early inner retinal dysfunction in streptozotocin-induced diabetic rats. AB - PURPOSE: Diabetes is known to alter retinal function, as measured with the electroretinogram (ERG), which shows a propensity toward inner retinal oscillatory potential (OPs) abnormalities. However, the effect that diabetes has on other ganglion cell-related responses is not known. This study was a systematic evaluation of streptozotocin (STZ) diabetes-related ERG changes in rats for the first 11 weeks after diabetogenesis. METHODS: Thirty Sprague-Dawley rats were randomly assigned to treated (50 mg/kg STZ (n = 16) and control groups (1 mL/kg citrate buffer, n = 14) at 6 weeks of age. Two control animals and four STZ animals were excluded because of blood glucose criteria or systemic complications. Diabetic animals were given daily SC injections of 1 to 2 units of long-acting insulin. ERGs were measured at 4, 8, and 11 weeks after treatment. The a-wave was used as an index of outer retinal function, whereas the b-wave, OPs, and the scotopic threshold response (STR) were used as indices of inner retinal function. RESULTS: Photoreceptoral (a-wave) and bipolar cell (b-wave) responses were not significantly reduced by STZ treatment. OPs were significantly reduced by 8 weeks (-25% +/- 7%, P < 0.05). The most severely affected component was the ganglion cell-dominated positive STR, which was significantly decreased from the first time point (-51% +/- 11% at 4 weeks, P < 0.05), but the negative component was unaffected over the 11-week period. CONCLUSIONS: The ganglion cell dominated pSTR showed large losses in STZ treated rats. PMID- 18421078 TI - ERK/p44p42 mitogen-activated protein kinase mediates EGF-stimulated proliferation of conjunctival goblet cells in culture. AB - PURPOSE: To determine whether activation of the ERK pathway by EGF leads to rat and human goblet cell proliferation. METHODS: The conjunctiva was removed from male Sprague-Dawley rats. Human conjunctiva was removed during ocular surgery. The tissue was minced and goblet cells were grown. The cells were stimulated with EGF (10(-7) M) for 1 and 5 minutes and Western blot analysis was performed with an antibody against phosphorylated EGFR, to measure the activation of the EGF receptor (EGFR). The cells were incubated with EGF (10(-7) M) for 24 hours, and cell proliferation was measured by WST-8. Inhibitors were added either 20 minutes before EGF or 2 hours after. The cells were stimulated with EGF (10(-7) M) for 1 minute to 24 hours. The number of cells expressing phosphorylated ERK (pERK) in the nucleus and Ki-67 was determined by immunofluorescence. RESULTS: EGF increased the activation of EGFR in rat conjunctival goblet cells. EGF-stimulated proliferation was inhibited by the EGFR inhibitor AG1478 and the MEK inhibitor U0126 in rat and human cultured goblet cells. EGF caused the translocation of pERK to the nucleus in a biphasic manner. Inhibition of the second peak with U0126 prevented proliferation. EGF-stimulated goblet cells progressed through the cell cycle expressing pERK in the nucleus. CONCLUSIONS: EGF stimulated human and rat conjunctival goblet cell proliferation by activating the EGFR. EGFR stimulated ERK causing its biphasic translocation to the nucleus. The second peak response is responsible for cell proliferation, but the role of the first peak is not known. PMID- 18421079 TI - Brightness, contrast, and color balance of digital versus film retinal images in the age-related eye disease study 2. AB - PURPOSE: To analyze brightness, contrast, and color balance of digital versus film retinal images in a multicenter clinical trial, to propose a model image from exemplars, and to optimize both image types for evaluation of age-related macular degeneration (AMD). METHODS: The Age-Related Eye Disease Study 2 (AREDS2) is enrolling subjects from 90 clinics, with three quarters of them using digital and one quarter using film cameras. Image brightness (B), contrast (C), and color balance (CB) were measured with three-color luminance histograms. First, the exemplars (film and digital) from expert groups were analyzed, and an AMD oriented model was constructed. Second, the impact of B/C/CB on the appearance of typical AMD abnormalities was analyzed. Third, B/C/CB in AREDS2 images were compared between film (156 eyes) and digital (605 eyes), and against the model. Fourth, suboptimal images were enhanced by adjusting B/C/CB to bring them into accord with model parameters. RESULTS: Exemplar images had similar brightness, contrast, and color balance, supporting an image model. Varying a specimen image through a wide range of B/C/CB revealed greatest contrast of drusen and pigment abnormalities against normal retinal pigment epithelium with the model parameters. AREDS2 digital images were more variable than film, with lower correspondence to our model. Ten percent of digital were too dim and 19% too bright (oversaturated), versus 1% and 4% of film, respectively. On average, digital had lower green channel contrast (giving less retinal detail) than film. Overly red color balance (weaker green) was observed in 23% of digital versus 8% of film. About half of digital (but fewer film) images required enhancement before AMD grading. After optimization of both image types, AREDS2 image quality was judged as good as that in AREDS (all film). CONCLUSIONS: A histogram-based model, derived from exemplars, provides a pragmatic guide for image analysis and enhancement. In AREDS2, the best digital images matched the best film. Overall, however, digital provided lower contrast of retinal detail. Digital images taken with higher G-to-R ratio showed better brightness and contrast management. Optimization of images in the multicenter study helps standardize documentation of AMD (ClinicalTrials.gov NCT00345176). PMID- 18421080 TI - Lovastatin inhibits TGF-beta-induced myofibroblast transdifferentiation in human tenon fibroblasts. AB - PURPOSE: The transdifferentiation of Tenon fibroblasts to myofibroblasts is a pivotal step in filtering bleb scarring. It is mediated by the cytokine TGF-beta, Rho-dependent contractility, and cell-matrix interactions in an interdependent fashion. HMG-CoA-reductase inhibitors (statins) have been shown to inhibit Rho GTPase signaling; therefore, the authors studied the influence of lovastatin on TGF-beta-mediated myofibroblast transdifferentiation to assess the potential use of statins in wound healing modulation. METHODS: Human Tenon fibroblasts were grown in culture, pretreated with lovastatin, lovastatin and mevalonate, or specific inhibitors of farnesyl transferase or geranylgeranyl transferase and were stimulated with TGF-beta1. alpha-Smooth muscle actin (alpha-SMA) and connective tissue growth factor (CTGF) transcription were assessed by real-time PCR. alpha-SMA protein expression and localization were studied by Western blot and confocal immunofluorescence microscopy. Cell contractility was determined in collagen gel contraction assays. Phosphorylation of the signaling proteins Smad 2/3 and p38 were detected by Western blot, and Smad-2/3 localization was determined by confocal immunofluorescence microscopy. RESULTS: Lovastatin inhibited TGF-beta-induced CTGF transcription, alpha-SMA expression and incorporation into actin stress fibers, and subsequent collagen gel contraction. These effects were reversed by mevalonate. The inhibition of geranylgeranyl transferase but not farnesyl transferase blocked TGF-beta-induced alpha-SMA expression. Lovastatin decreased TGF-beta-induced p38 activation, whereas Smad 2/3 phosphorylation and nuclear translocation were preserved. CONCLUSIONS: Lovastatin inhibits TGF-beta-induced myofibroblast transdifferentiation in human Tenon fibroblasts, most likely by interfering with Rho-signaling. Statins may, therefore, serve to inhibit scarring after filtering glaucoma surgery. PMID- 18421081 TI - Neuroprotective effect of intravitreal triamcinolone acetonide against photoreceptor apoptosis in a rabbit model of subretinal hemorrhage. AB - PURPOSE: To study photoreceptor apoptosis and iron migration as mechanisms of retinotoxicity in a rabbit model of subretinal hemorrhage (SRH) and to assess intravitreal triamcinolone acetonide (IVTA) for anti-apoptotic and neuroprotective effects. METHODS: In adult rabbits, eyes were studied histologically after subretinal injection of autologous blood. For comparisons of control eyes with eyes injected with 2 mg IVTA, morphometric analysis was performed with light microscopy, whereas apoptosis was quantified with terminal dUTP nick end labeling (TUNEL) and fluorescence microscopy. Localization of retinal iron was assessed with Perls' stain. RESULTS: Photoreceptor degeneration was initiated 48 hours after exposure to subretinal blood and progressed over 7 days. Increased TUNEL positivity demonstrating apoptotic cell death was associated with SRH and photoreceptor loss. VIP-Perls staining demonstrated iron in the photoreceptor layer and retinal pigment epithelium that correlated with photoreceptor degeneration. Treatment with IVTA enhanced photoreceptor cell survival by 11% at 48 hours and by 45% at 72 hours (P = 0.01) and reduced photoreceptor apoptosis ratios by 25% at 48 hours (P = 0.006). CONCLUSIONS: Photoreceptor toxicity caused by SRH occurs at least in part by apoptosis and is associated with iron migration to the photoreceptor layer. Treatment with IVTA reduced photoreceptor loss and apoptosis, indicating a neuroprotective action. Therapies to target SRH may augment anti-VEGF treatments in exudative age-related macular degeneration and other diseases of choroidal neovascularization. PMID- 18421082 TI - Visualization of in situ intracellular aggregation of two cataract-associated human gamma-crystallin mutants: lose a tail, lose transparency. AB - PURPOSE: To understand the molecular features underlying autosomal dominant congenital cataracts caused by the deletion mutations W156X in human gammaD crystallin and W157X in human gammaC-crystallin. METHODS: Normal and mutant cDNAs (with the enhanced green fluorescent protein [EGFP] tag in the front) were cloned into the pEGFP-C1 vector, transfected into various cell lines, and observed under a confocal microscope for EGFP fluorescence. Normal and W156X gammaD cDNAs were also cloned into the pET21a(+) vector, and the recombinant proteins were overexpressed in the BL-21(DE3)pLysS strain of Escherichia coli, purified, and isolated. The conformational features, structural stability, and solubility in aqueous solution of the mutant protein were compared with those of the wild type using spectroscopic methods. Comparative molecular modeling was performed to provide additional structural information. RESULTS: Transfection of the EGFP tagged mutant cDNAs into several cell lines led to the visualization of aggregates, whereas that of wild-type cDNAs did not. Turning to the properties of the expressed proteins, the mutant molecules show remarkable reduction in solubility. They also seem to have a greater degree of surface hydrophobicity than the wild-type molecules, most likely accounting for self-aggregation. Molecular modeling studies support these features. CONCLUSIONS: The deletion of C terminal 18 residues of human gammaC- and gammaD-crystallins exposes the side chains of several hydrophobic residues in the sequence to the solvent, causing the molecule to self-aggregate. This feature appears to be reflected in situ on the introduction of the mutants in human lens epithelial cells. PMID- 18421083 TI - Retinal nerve fiber layer analysis in RP patients using Fourier-domain OCT. AB - PURPOSE: To determine peripapillary retinal nerve fiber layer thickness (RNFL) abnormalities in patients with retinitis pigmentosa (RP) using Fourier-domain optical coherence tomography (Fd-OCT) and to evaluate the potential effect of cystoid macular edema (CME) or axial length on RNFL measurements in such patients. METHODS: Ninety-seven eyes of 52 patients with diagnoses of retinitis pigmentosa or Usher syndrome type II underwent complete ocular examination. Peripapillary RNFL thickness was measured using Fd-OCT in 16 segments from 4 quadrants--temporal (316 degrees -45 degrees ), superior (46 degrees -135 degrees ), nasal (136 degrees -225 degrees ), and inferior (226 degrees -315 degrees ). These measurements were compared with age- and disc size-adjusted control values. Further analyses were performed to determine the correlation of axial length or CME with RNFL thickness. RESULTS: Thinning of the RNFL was observed in 37 eyes (38.14%) of 23 patients (44.23%). A maximum number of eyes had thinning in the nasal quadrant followed by the inferior quadrant; the superior and temporal quadrants were abnormally thin in fewer eyes. No correlation was found between axial length and RNFL thickness in the total cohort (correlation coefficient, 0.039). An abnormal increase in RNFL thickness was observed in 21.65% eyes, but no association was found between the presence of CME and increased RNFL thickness. CONCLUSIONS: RP eyes may show abnormal thinning or increased thickness of RNFL measurements on testing with Fd-OCT. RNFL defects observed by OCT testing document the presence of anatomic defects in more anterior structures within the retina in a notable number of patients with RP. PMID- 18421084 TI - Factors influencing the experience of oscillopsia in infantile nystagmus syndrome. AB - PURPOSE: Perceptual instability in infantile nystagmus syndrome (INS) has been reported occasionally. This study was conducted to examine the factors that influence perceptual stability in 18 individuals with INS. METHODS: The subjects were instructed to look continuously at a fixation LED centered in an image (38 degrees x 32 degrees ) at two luminance levels (3.25 and 0.46 cd/m(2), with 21% and 96% contrast, respectively) throughout all trials. A trial consisted of the fixation LED on, followed by a peripheral LED on, and then both LEDs off. Subjects then reported what they perceived. Five trials were conducted per contrast image. Eye movements were recorded with a limbal tracker. After testing, each subject completed a questionnaire to determine whether they ever had or were presently experiencing oscillopsia. RESULTS: Sixteen of 18 subjects reported experiencing oscillopsia on the questionnaire. In the laboratory, the percentages of trials with perceptions of motion of the LED and background were as follows: neither, 45% to 60%; background only, 15% to 30%; both, approximately 15%; and LED only, approximately 10%. Over all trials, 14/18 and 13/17 subjects experienced oscillopsia for the low- and high-contrast images, respectively (i.e., four subjects never experienced oscillopsia). The background was frequently seen moving when both images were displayed, regardless of contrast and/or condition. Trials with and without oscillopsia did not differ between the foveation periods. CONCLUSIONS: Subjects with INS may experience spatially inhomogeneous oscillopsia under certain viewing conditions. The physical attributes of the stimulus, repeated trials, different conduction times, and the role of divided attention may influence a subject's perception differently. PMID- 18421085 TI - Photoreceptor protection after photodynamic therapy using dexamethasone in a rat model of choroidal neovascularization. AB - PURPOSE: To study whether corticosteroids protect photoreceptors when combined with photodynamic therapy (PDT) in a laser-induced model of choroidal neovascularization (CNV). METHODS: PDT was performed in 36 Brown-Norway rats 2 weeks after laser induction of CNV. The expressional change of several cytokines and chemokines in the CNV lesions after PDT was measured by real-time PCR in combination with laser-capture microdissection. Immunostaining for monocyte chemoattractant protein (MCP)-1, C-C chemokine receptor 2(CCR2), interleukin (IL) 1beta, and myeloperoxidase(MPO) were performed. To study the effect of corticosteroids in combination with PDT, either dexamethasone (100 mg/kg) or control was injected intraperitoneally 1 hour before PDT. Animals were killed 24 hours or 1 week after PDT. CNV was examined by fluorescein angiography and choroidal flatmount. Photoreceptor degeneration was evaluated by TUNEL assay. RESULTS: MCP-1 and IL-1beta was increased in CNV lesions 24 hours after PDT. CCR2 was also expressed in laser-induced CNV but did not increase after PDT. Twenty four hours after PDT, MPO-positive cells were noted in the CNV lesions. Dexamethasone-treated animals had significantly fewer TUNEL-positive cells in the photoreceptor layer than did the control animals (P < 0.05) after PDT. Fluorescein angiographic grading of CNV closure 6 days after PDT showed a closure rate in the dexamethasone-treated group of 31% (15/48 lesions) compared to 10% (4/42 lesions) in the control group (P < 0.05). CNV size was significantly smaller in the dexamethasone-treated group 1 week after PDT compared with the control (P < 0.05). CONCLUSIONS: Systemic administration of dexamethasone combined with PDT reduces photoreceptor apoptosis, increases angiographic closure, and reduces CNV size compared with PDT alone in a rat model. PMID- 18421086 TI - PDGF- and insulin/IGF-1-specific distinct modes of class IA PI 3-kinase activation in normal rat retinas and RGC-5 retinal ganglion cells. AB - PURPOSE: To compare PDGF- and insulin/IGF-1-induced class I(A) PI 3-kinase/Akt survival signaling in normal retinas and retinal ganglion cells (RGCs). METHODS: Normal rat retinas and RGC-5 cells were used for (1) immunohistochemical and immunoblot studies to detect PDGF receptor (PDGFR) subtypes and (2) immunoprecipitation, immunoblot, and in vitro lipid kinase assays to determine basal and PDGF-induced class I(A) PI 3-kinase/Akt survival signaling, in comparison with insulin or IGF-1 responses. Furthermore, RGC-5 cells were exposed to broad-spectrum (LY294002) or p110 isoform-selective (PI-103) PI 3-kinase inhibitors (versus Akt inhibitor) to assess the consequent effects on Akt phosphorylation, caspase-3/PARP cleavage, apoptotic phenotype, and cell viability, as a function of serum trophic factors. RESULTS: PDGFR-alpha and -beta immunoreactivity was observed in rat retinal Muller cells and in the RGC layer and blood vessels, respectively. In addition, PDGFR-alpha and -beta protein expression was observed in RGC-5 cells. Both retinas and RGC-5 cells exhibited a similar pattern of subunit-specific basal class I(A) PI 3-kinase activity, which was stimulated in a temporal and signal-specific manner by PDGF and insulin/IGF 1. Furthermore, RGC-5 cells showed PDGFR-alpha/beta tyrosine phosphorylation that induced the p85alpha regulatory subunit to activate p110alpha/beta-associated class I(A) PI 3-kinase, which in turn enhanced Akt phosphorylation. Exposure of serum-deprived RGC-5 cells to PI 3-kinase or Akt inhibitors increased susceptibility to apoptotic phenotype as revealed by caspase-3 and PARP cleavage. CONCLUSIONS: The present findings provide direct evidence of two distinct modes of retinal class I(A) PI 3-kinase activation that occurs in response to PDGF receptor and insulin/IGF-1 receptor stimulation. PDGF-induced PI 3 kinase/PIP3/Akt axis may provide new therapeutic approaches to ameliorate cell death in diabetic retinopathy and other retinal neurodegenerations. PMID- 18421087 TI - Multiple gene polymorphisms in the complement factor h gene are associated with exudative age-related macular degeneration in chinese. AB - PURPOSE: Variants in the complement factor H (CFH) gene have been shown to be strongly associated with age-related macular degeneration (AMD). In this study, sequence alterations in CFH were investigated in 163 Chinese patients with exudative AMD and 155 unrelated Chinese control subjects. METHODS: All the 22 CFH exons, intron-exon boundaries, and promoter sequences were screened by polymerase chain reaction and DNA sequencing. RESULTS: Fifty-eight sequence changes, 42 of them novel, were identified. Six SNPs with an allele frequency >30% were significantly associated with exudative AMD. SNP rs3753396 was novel; the rest had been reported: rs3753394, rs551397, rs800292, rs2274700, and rs1329428. Two haplotype blocks were constructed. The TG haplotype for rs551397 and rs800292 was the major haplotype that conferred a significantly increased susceptibility to exudative AMD (P(corr) = 0.0001, OR = 1.91, 95% CI = 1.36-2.68). CONCLUSIONS: The findings support prior evidence that the CFH gene is one of the AMD-associated genes. There is a different distribution pattern of CFH variants in the Chinese compared with other populations. Individual SNP and haplotype analyses revealed that the ancient alleles at the 5' end of CFH contribute to an increased susceptibility to exudative AMD. PMID- 18421089 TI - Diminished frequency and function of CD4+CD25high regulatory T cells associated with active uveitis in Vogt-Koyanagi-Harada syndrome. AB - PURPOSE: CD4(+)CD25(high) regulatory T (Treg) cells have been shown to be involved in the pathogenesis of autoimmune diseases. Vogt-Koyanagi-Harada (VKH) syndrome is an organ-specific autoimmune disease. This study was designed to phenotypically and functionally characterize peripheral blood CD4(+)CD25(high) Treg cells in VKH patients with active uveitis. METHODS: Blood samples were taken from 30 patients with active VKH, 19 patients with inactive VKH, and 26 healthy controls. Peripheral blood mononuclear cells were subjected to flow cytometry for analysis of phenotypes of the CD4(+)CD25(high) Treg cells. For functional analysis, CD4(+)CD25(high) Treg cells and CD4(+)CD25(-) T cells were separated by means of magnetic-assisted cell sorting and subsequently cocultured for 6 days. The proliferation of CD4(+)CD25(-) T cells was measured by [(3)H] thymidine incorporation assay. The levels of IFN-gamma, IL-17, and IL-13 in the supernatants were determined by enzyme-linked immunosorbent assay. RESULTS: Significantly decreased frequencies of CD4(+)CD25(high) Treg cells and percentages of FOXP3(+) cells in these Treg cells were shown in patients with active VKH. Treg cells from patients with active VKH showed a significant deficiency in suppressing the proliferation of CD4(+)CD25(-) T cells and inhibiting the production of IFN-gamma and IL-13 by CD4(+)CD25(-) T cells. CD4(+)CD25(high) Treg cells from VKH patients or healthy controls did not markedly inhibit or promote IL-17 production. CONCLUSIONS: A significantly decreased frequency and diminished function of CD4(+)CD25(high) Treg cells is associated with active uveitis in patients with VKH syndrome. These results suggest that these dysfunctional CD4(+)CD25(high) Treg cells may play a role in the pathogenesis of uveitis in VKH syndrome. PMID- 18421088 TI - Inhibition of crystallin ascorbylation by nucleophilic compounds in the hSVCT2 mouse model of lenticular aging. AB - PURPOSE: Senile cataracts are associated with oxidation, fragmentation, cross linking, insolubilization, and yellow pigmentation of lens crystallins. This process is partially explained by advanced glycation end products (AGEs) from ascorbic acid (ASA), as the authors unequivocally demonstrated in an hSVCT2 transgenic mouse. The authors present the first pharmacologic intervention study against ascorbylation in these mice. METHODS: Five groups of mice from 2 to 9 months of age (10 mice/group) were fed a diet containing 0.1% (wt/wt) aminoguanidine, pyridoxamine, penicillamine, and nucleophilic compounds NC-I and NC-II. AGEs were determined in crystallin digests using high-performance liquid chromatography, liquid chromatography-mass spectrometry, or gas chromatography mass spectrometry. Lens protein extract was incubated in vitro with ASA or dehydroascorbic acid. RESULTS: The ASA level increased approximately 10-fold in all groups and was unaffected by treatment. AGEs were increased several-fold in transgenic compared with control lenses. Body weight, food intake, lenticular glutathione, and glycated lysine level were unaltered. In vitro, all compounds inhibited AGE formation. In vivo, NC-I and NC-II significantly decreased protein fluorescence at lambda(ex)335/(em)385 (P = 0.045, P = 0.017, respectively) and lambda(ex)370/(em)440 (P = 0.029, P = 0.007, respectively). Other inhibitors had no effect. After 7 months, only NC-I and NC-II induced a 50% reduction in pentosidine (P = NS for NC-I; P = 0.035 for NC-II). NC-I also decreased carboxymethyllysine (P = 0.032) and carboxyethyllysine (P = NS). Fluorescent cross-link K2P was decreased by NC-I, NC-II, aminoguanidine, and pyridoxamine (P = NS). CONCLUSIONS: Pharmacologically blocking protein ascorbylation with absorbable guanidino compounds is feasible and may represent a new strategy for the delay of age-related nuclear sclerosis of the lens. PMID- 18421090 TI - Neural rim characteristics of healthy South Indians: the Chennai Glaucoma Study. AB - PURPOSE: To report neural rim dimensions for South Indians and examine rim shape with relevance to clinical evaluation. METHODS: Healthy phakic participants (n = 623) of the Chennai Glaucoma Study with normal frequency-doubling perimetry underwent complete eye examinations including optic disc digital stereophotography. Planimetry was performed under stereoviewing conditions using custom software. Rim area, shape, and associations were examined. Rim area asymmetry was studied in a subgroup of 565 subjects. RESULTS: Mean neuroretinal rim area was 2.29 +/- 0.39 mm(2). Disc area (P < 0.001) and type of cupping (P < 0.001) were associated with rim area. Mean rim area asymmetry was 0.18 +/- 0.15 mm(2); 95% of subjects had asymmetry < 0.5 mm(2). Disc area asymmetry (0.359, P < 0.0001) and intraocular pressure (IOP) asymmetry (P = 0.004) were related to rim area asymmetry. On average, the inferior rim was thickest and the temporal was thinnest. Mean inferior by superior rim width was 1.18 +/- 0.17; 2.5 percentile, 0.9. Thirty-eight (7.1%) subjects had the superior rim thicker than the inferior rim, the occurrence of which was associated with disc torsion (P = 0.002) and male sex (P = 0.04). Of the clinically relevant rim width measures in glaucoma (i.e., inferior, superior, and temporal) the temporal rim was thinnest in 469 (87.8%) eyes. Horizontally oval disc shape (P < 0.0001), type of cupping (P = 0.006), and astigmatism (P = 0.001) were associated with the presence of thicker temporal than superior/inferior rims. CONCLUSIONS: The report provides hitherto unreported neural rim measurements among healthy South Indians. The ISNT rule (inferior rim thicker than superior rim, thicker than nasal rim, thicker than temporal rim) was violated in a significant minority. Physiological associations with such violations are described. PMID- 18421091 TI - Characterization of a spontaneous mouse retinal pigment epithelial cell line B6 RPE07. AB - PURPOSE: A spontaneously arising retinal pigment epithelial (RPE) cell line (B6 RPE07) was cloned from a primary culture of mouse RPE cells and maintained in culture for more than 18 months. Morphologic and functional properties of this cell line have been characterized. METHODS: The morphology of the B6-RPE07 cells was examined by phase-contrast light microscopy, electron microscopy, and confocal microscopy. Barrier properties were measured by the flux of fluorescence from the apical to the basolateral compartment of culture chambers. The abilities of the cells to bind/phagocytose photoreceptor outer segments (POS) were determined by confocal microscopy, electron microscopy, and flow cytometry. Cytokine/chemokine secretion was measured by cytometric bead array. The expression of visual cycle proteins was determined by RT-PCR and Western blotting. RESULTS: In standard culture conditions, B6-RPE07 cells display cobblestone morphology. When cultured on three-dimensional (3D) collagen gel coated membranes, B6-RPE07 cells exhibit a monolayer epithelial polarization with apical surface microvilli. Immunohistochemistry of B6-RPE07 cultures revealed a high expression of pan-cytokeratin. B6-RPE07 cells also expressed the retinal pigment epithelium-specific marker CRALBP, but not RPE65. Cell junction proteins ZO-1 and beta-catenin, but not claudin-1/3 or occludin-1, were observed in B6 RPE07 cells. B6-RPE07 cells are able to bind, phagocytose, and digest POS. Finally, B6-RPE07 cells produce high levels of IL-6 and CCL2. CONCLUSIONS: This is the first report of a mouse RPE cell line with morphology, phenotype, and function similar to those of in vivo mouse RPE cells. This cell line will be a valuable resource for future RPE studies, in particular for in vivo gene modification and transplantation studies. PMID- 18421092 TI - Angiopoietin-like 7 secretion is induced by glaucoma stimuli and its concentration is elevated in glaucomatous aqueous humor. AB - PURPOSE: To investigate the possibility that Angiopoietin-like 7 (ANGPTL7) protein is involved in the pathogenesis of glaucoma. METHODS: Primary human trabecular meshwork (TM) cells and corneoscleral explants were stimulated with either dexamethasone (DEX) or transforming growth factor beta (TGFbeta), and ANGPTL7 protein secreted into culture medium was determined by Western blot analysis. The effect of stable overexpression of ANGPTL7 in transfected immortalized TM cell lines on collagen expression was investigated by immunocytochemistry. Localization of ANGPTL7 protein in human eyes was determined by immunohistochemistry. The concentration of ANGPTL7 protein in aqueous humor (AH) from patients with glaucoma and control patients was compared by Western blot analysis. The beagle model of primary open-angle glaucoma (POAG) was used to correlate ANGPTL7 protein levels in canine AH with disease progression. RESULTS: TGFbeta and DEX stimulated secretion of ANGPTL7 protein by TM cells and corneoscleral explants. Overexpression of ANGPTL7 by immortalized TM cell lines increased expression of type I collagen. Expression of ANGPTL7 protein was located in the corneal stroma, near the limbus, and throughout the sclera, with lower expression in the TM. In the lamina cribrosa, ANGPTL7 expression was associated with the cribriform plates. The concentration of ANGPTL7 protein was elevated in AH from patients with glaucoma and increased as disease progressed in POAG beagle dogs. CONCLUSIONS: Induction of ANGPTL7 secretion by glaucoma stimuli and increased concentration of ANGPTL7 in glaucomatous AH suggest that ANGPTL7 is overexpressed in glaucoma. Since overexpression of ANGPTL7 increases collagen expression, a potential disease mechanism, ANGPTL7 could have a pathogenic role in glaucoma, and may serve as a potential therapeutic target. PMID- 18421093 TI - Transcriptional regulation of the human alpha6 integrin gene by the transcription factor NFI during corneal wound healing. AB - PURPOSE: Wound healing of the corneal epithelium is highly influenced by regulation of integrin gene expression. A recent study demonstrated that laminin (LM), a major constituent of the extracellular matrix (ECM), reduces expression of the human alpha6 integrin subunit gene by altering the properties of the transcription factor (TF) Sp1. In this work, a target site was identified for the TF nuclear factor I (NFI) on the human alpha6 gene, and its regulatory influence was characterized in corneal epithelial cells. METHODS: Plasmids bearing the alpha6 promoter fused to the CAT gene were transfected into human (HCECs) and rabbit (RCECs) corneal epithelial cells grown on LM. The DNA-binding site for NFI in the alpha6 promoter was identified by DNase I footprinting. Expression and DNA binding of NFI was monitored by Western blot, RT-PCR, and electrophoretic mobility shift assays (EMSAs), and its function was investigated through RNAi and NFI overexpression assays. RESULTS: All NFI isoforms were found to be expressed in HCECs and RCECs. Transfection analyses revealed that NFI is a repressor of alpha6 expression in both types of cells. LM increases expression of NFI, whereas inhibition of each NFI isoform increases promoter activity suggesting that NFI is a key repressor of alpha6 transcription. In addition, the negative influence of NFI appears to be potentiated by the degradation of Sp1 when cells are grown on LM. CONCLUSIONS: Repression of alpha6 expression therefore contributes to the final steps of corneal wound healing by both reducing proliferation and allowing attachment of the epithelium to the basal membrane. PMID- 18421094 TI - Blood levels of vitamin C, carotenoids and retinol are inversely associated with cataract in a North Indian population. AB - PURPOSE: To examine the association of blood antioxidants with cataract. METHODS: Cross-sectional study of people aged >or=50 years identified from a household enumeration of 11 randomly sampled villages in North India. Participants were interviewed for putative risk factors (tobacco, alcohol, biomass fuel use, sunlight exposure, and socioeconomic status) and underwent lens photography and blood sampling. Lens photographs (nuclear, cortical, and posterior subcapsular) were graded according to the Lens Opacities Classification System (LOCS II). Cataract was defined as LOCS II grade >or=2 for any opacity or ungradable, because of dense opacification or history of cataract surgery. People without cataract were defined as LOCS II <2 on all three types of opacity, with absence of previous surgery. RESULTS: Of 1443 people aged >or=50 years, 94% were interviewed, 87% attended an eye examination, and 78% gave a blood sample; 1112 (77%) were included in the analyses. Compared with levels in Western populations, antioxidants were low, especially vitamin C. Vitamin C was inversely associated with cataract. Odds ratios (OR) for the highest (>or=15 micromol/L) compared with the lowest ( 1 for PBC reached 25% in DIBH and about 15% in FB. CC and AAA performed, in contrast, similarly and with fractions of lung voxels with gamma > 1 in average inferior to 2% in FB and 4-5% (AAA) or 6-8% (CC) in DIBH. In summary, PBC proved to be severely defective in calculations involving lungs and particularly for cases where specific respiratory phases (e.g. DIBH) are assumed for treatment. In contrast, CC and AAA manifested a high degree of consistency against the Monte Carlo method and provided stable results over the entire range of clinically relevant densities. PMID- 18421118 TI - Time-dependent cell disintegration kinetics in lung tumors after irradiation. AB - We study the time-dependent disintegration kinetics of tumor cells that did not survive radiotherapy treatment. To evaluate the cell disintegration rate after irradiation, we studied the volume changes of solitary lung tumors after stereotactic radiotherapy. The analysis is performed using two approximations: (1) tumor volume is a linear function of the total cell number in the tumor and (2) the cell disintegration rate is governed by the exponential decay with constant risk, which is defined by the initial cell number and a half-life T(1/2). The half-life T(1/2) is determined using the least-squares fit to the clinical data on lung tumor size variation with time after stereotactic radiotherapy. We show that the tumor volume variation after stereotactic radiotherapy of solitary lung tumors can be approximated by an exponential function. A small constant component in the volume variation does not change with time; however, this component may be the residual irregular density due to radiation fibrosis and was, therefore, subtracted from the total volume variation in our computations. Using computerized fitting of the exponent function to the clinical data for selected patients, we have determined that the average half life T(1/2) of cell disintegration is 28.2 days for squamous cell carcinoma and 72.4 days for adenocarcinoma. This model is needed for simulating the tumor volume variation during radiotherapy, which may be important for time-dependent treatment planning of proton therapy that is sensitive to density variations. PMID- 18421119 TI - Neutron-stimulated emission computed tomography of a multi-element phantom. AB - This paper describes the implementation of neutron-stimulated emission computed tomography (NSECT) for non-invasive imaging and reconstruction of a multi-element phantom. The experimental apparatus and process for acquisition of multi-spectral projection data are described along with the reconstruction algorithm and images of the two elements in the phantom. Independent tomographic reconstruction of each element of the multi-element phantom was performed successfully. This reconstruction result is the first of its kind and provides encouraging proof of concept for proposed subsequent spectroscopic tomography of biological samples using NSECT. PMID- 18421120 TI - Effect of inter-crystal scatter on estimation methods for random coincidences and subsequent correction. AB - Random coincidences can contribute substantially to the background in positron emission tomography (PET). Several estimation methods are being used for correcting them. The goal of this study was to investigate the validity of techniques for random coincidence estimation, with various low-energy thresholds (LETs). Simulated singles list-mode data of the MADPET-II small animal PET scanner were used as input. The simulations have been performed using the GATE simulation toolkit. Several sources with different geometries have been employed. We evaluated the number of random events using three methods: delayed window (DW), singles rate (SR) and time histogram fitting (TH). Since the GATE simulations allow random and true coincidences to be distinguished, a comparison between the number of random coincidences estimated using the standard methods and the number obtained using GATE was performed. An overestimation in the number of random events was observed using the DW and SR methods. This overestimation decreases for LETs higher than 255 keV. It is additionally reduced when the single events which have undergone a Compton interaction in crystals before being detected are removed from the data. These two observations lead us to infer that the overestimation is due to inter-crystal scatter. The effect of this mismatch in the reconstructed images is important for quantification because it leads to an underestimation of activity. This was shown using a hot-cold-background source with 3.7 MBq total activity in the background region and a 1.59 MBq total activity in the hot region. For both 200 keV and 400 keV LET, an overestimation of random coincidences for the DW and SR methods was observed, resulting in approximately 1.5% or more (at 200 keV LET: 1.7% for DW and 7% for SR) and less than 1% (at 400 keV LET: both methods) underestimation of activity within the background region. In almost all cases, images obtained by compensating for random events in the reconstruction algorithm were better in terms of quantification than the images made with precorrected data. PMID- 18421121 TI - Uncertainty and bias in contrast concentration measurements using spoiled gradient echo pulse sequences. AB - Dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) is a widely used technique for assessing tissue physiology. Spoiled gradient echo (SPGR) pulse sequences are one of the most common methods for acquisition of DCE-MRI data, providing high temporal and spatial resolution with strong T(1)-weighting. Conversion of SPGR signal to concentration is briefly reviewed, and a new closed form expression for concentration measurement uncertainty for finite signal-to noise ratio (SNR) and baseline scan time is derived. This result is applicable to arbitrary concentration-dependent relaxation rate and is valid over the same domain as the theoretical SPGR signal equation. Expressions for the lower and upper bounds on measurable concentration are also derived. The existence of a concentration- and tissue-dependent optimal flip angle that minimizes concentration uncertainty is demonstrated and it is shown that, for clinically relevant pulse sequence parameters, this optimal flip angle is significantly larger than the corresponding Ernst angle. Analysis of three pulse sequences from the DCE-MRI literature shows that optimization of flip angle using the methods discussed here leads to potential improvements of 10-1166% in effective SNR over the 0.5-5.0 mM concentration range with minimal or no loss of measurement accuracy down to 0.1 mM. In vivo data from three study patients provide further support for our theoretical expression for concentration measurement uncertainty, with predicted and experimental estimates agreeing to within +/- 30%. Equations for concentration bias resulting from biases in flip angle and from pre-contrast relaxation time and contrast relaxivity (both longitudinal and transverse) are also derived in closed-form. The resulting equations show the potential for significant contributions to bias in concentration measurement arising from even relatively small mis-specification of flip angle and/or pre-contrast longitudinal relaxation time, particularly at high contrast concentrations. PMID- 18421122 TI - Can megavoltage computed tomography reduce proton range uncertainties in treatment plans for patients with large metal implants? AB - Treatment planning calculations for proton therapy require an accurate knowledge of radiological path length, or range, to the distal edge of the target volume. In most cases, the range may be calculated with sufficient accuracy using kilovoltage (kV) computed tomography (CT) images. However, metal implants such as hip prostheses can cause severe streak artifacts that lead to large uncertainties in proton range. The purposes of this study were to quantify streak-related range errors and to determine if they could be avoided by using artifact-free megavoltage (MV) CT images in treatment planning. Proton treatment plans were prepared for a rigid, heterogeneous phantom and for a prostate cancer patient with a metal hip prosthesis using corrected and uncorrected kVCT images alone, uncorrected MVCT images and a combination of registered MVCT and kVCT images (the hybrid approach). Streak-induced range errors of 5-12 mm were present in the uncorrected kVCT-based patient plan. Correcting the streaks by manually assigning estimated true Hounsfield units improved the range accuracy. In a rigid heterogeneous phantom, the implant-related range uncertainty was estimated at <3 mm for both the corrected kVCT-based plan and the uncorrected MVCT-based plan. The hybrid planning approach yielded the best overall result. In this approach, the kVCT images provided good delineation of soft tissues due to high-contrast resolution, and the streak-free MVCT images provided smaller range uncertainties because they did not require artifact correction. PMID- 18421123 TI - Mechanical characterization of articular cartilage by combining magnetic resonance imaging and finite-element analysis: a potential functional imaging technique. AB - Magnetic resonance imaging (MRI) provides a method for non-invasive characterization of cartilage composition and structure. We aimed to see whether T(1) and T(2) relaxation times are related to proteoglycan (PG) and collagen specific mechanical properties of articular cartilage. Specifically, we analyzed whether variations in the depthwise collagen orientation, as assessed by the laminae obtained from T(2) profiles, affect the mechanical characteristics of cartilage. After MRI and unconfined compression tests of human and bovine patellar cartilage samples, fibril-reinforced poroviscoelastic finite-element models (FEM), with depthwise collagen orientations implemented from quantitative T(2) maps (3 laminae for human, 3-7 laminae for bovine), were constructed to analyze the non-fibrillar matrix modulus (PG specific), fibril modulus (collagen specific) and permeability of the samples. In bovine cartilage, the non-fibrillar matrix modulus (R = -0.64, p < 0.05) as well as the initial permeability (R = 0.70, p < 0.05) correlated with T(1). In bovine cartilage, T(2) correlated positively with the initial fibril modulus (R = 0.62, p = 0.05). In human cartilage, the initial fibril modulus correlated negatively (R = -0.61, p < 0.05) with T(2). Based on the simulations, cartilage with a complex collagen architecture (5 or 7 laminae), leading to high bulk T(2) due to magic angle effects, provided higher compressive stiffness than tissue with a simple collagen architecture (3 laminae). Our results suggest that T(1) reflects PG-specific mechanical properties of cartilage. High T(2) is characteristic to soft cartilage with a classical collagen architecture. Contradictorily, high bulk T(2) can also be found in stiff cartilage with a multilaminar collagen fibril network. By emerging MRI and FEM, the present study establishes a step toward functional imaging of articular cartilage. PMID- 18421124 TI - Dual-energy CT-based material extraction for tissue segmentation in Monte Carlo dose calculations. AB - Monte Carlo (MC) dose calculations are performed on patient geometries derived from computed tomography (CT) images. For most available MC codes, the Hounsfield units (HU) in each voxel of a CT image have to be converted into mass density (rho) and material type. This is typically done with a (HU; rho) calibration curve which may lead to mis-assignment of media. In this work, an improved material segmentation using dual-energy CT-based material extraction is presented. For this purpose, the differences in extracted effective atomic numbers Z and the relative electron densities rho(e) of each voxel are used. Dual energy CT material extraction based on parametrization of the linear attenuation coefficient for 17 tissue-equivalent inserts inside a solid water phantom was done. Scans of the phantom were acquired at 100 kVp and 140 kVp from which Z and rho(e) values of each insert were derived. The mean errors on Z and rho(e) extraction were 2.8% and 1.8%, respectively. Phantom dose calculations were performed for 250 kVp and 18 MV photon beams and an 18 MeV electron beam in the EGSnrc/DOSXYZnrc code. Two material assignments were used: the conventional (HU; rho) and the novel (HU; rho, Z) dual-energy CT tissue segmentation. The dose calculation errors using the conventional tissue segmentation were as high as 17% in a mis-assigned soft bone tissue-equivalent material for the 250 kVp photon beam. Similarly, the errors for the 18 MeV electron beam and the 18 MV photon beam were up to 6% and 3% in some mis-assigned media. The assignment of all tissue-equivalent inserts was accurate using the novel dual-energy CT material assignment. As a result, the dose calculation errors were below 1% in all beam arrangements. Comparable improvement in dose calculation accuracy is expected for human tissues. The dual-energy tissue segmentation offers a significantly higher accuracy compared to the conventional single-energy segmentation. PMID- 18421125 TI - An investigation into structural changes due to deuteration. AB - Perdeuteration of proteins is becoming more commonplace and the assumption is in general that deuteration does not affect protein structure. In this work, the effect of deuteration on structure is examined by data mining, largely of the Cambridge Structural Database but also of the Inorganic Crystal Structure Database, for deuterated and hydrogenated pairs of small-molecule structures analysed by neutron and X-ray crystallography. Differences between these small molecule structures have been calculated and the results thus far follow the initial assumption. However, functional changes are known, e.g. D(2)O is toxic to living systems but H(2)O is not, kinetics change, small pH to pD changes occur, proteins stiffen in D(2)O and ferroelectrics alter their properties. PMID- 18421126 TI - Phase stability in ferroelectric bismuth titanate: a first-principles study. AB - Experimental data on the structure of ferroelectric oxide bismuth titanate suggest two different kinds of structures, i.e. orthorhombic and monoclinic. Density-functional-theory-based (DFT) first-principles calculations have been performed to determine the most stable structure of bismuth titanate among experimentally observed structures. Orthorhombic and monoclinic phases are optimized to zero pressure and lattice parameters were determined as a=5.4370, b=5.4260, c=32.6833 A and Z=4 for the structure with space group B2cb, and a=5.4289, b=5.4077, c=32.8762 A, beta=90.08 degrees and Z=4 for the structure with space group B1a1. Static and relaxation calculations show that the monoclinic structure with space group B1a1 is the most stable structure. PMID- 18421127 TI - Quasiperiodic plane tilings based on stepped surfaces. AB - Static and dynamic characteristics of layerwise growth in two-dimensional quasiperiodic Ito-Ohtsuki tilings are studied. These tilings are the projections of three-dimensional stepped surfaces. It is proved that these tilings have hexagonal self-similar growth with bounded radius of neighborhood. A formula is given for the averaged coordination number. Deviations of coordination numbers from its average are quasiperiodic. Ito-Ohtsuki tiling can be decomposed into one dimensional sector layers. These sector layers are one-dimensional quasiperiodic tilings with properties like Ito-Ohtsuki tilings. PMID- 18421128 TI - X-ray structure refinement using aspherical atomic density functions obtained from quantum-mechanical calculations. AB - An approach is outlined for X-ray structure refinement using atomic density fragments obtained by Hirshfeld partitioning of quantum-mechanical density fragments. Results are presented for crystal structure refinements of urea and benzene using these 'Hirshfeld atoms'. Using this procedure, the quantum mechanical non-spherical electron density is taken into account in the structural model based on the conformation found in the crystal. Contrary to current consensus in structure refinement, the anisotropic displacement parameters of H atoms can be reproduced from neutron diffraction measurements simply from a least squares fit using the Hirshfeld atoms derived from the BLYP level of theory and including a simple point-charge model to treat the crystal environment. PMID- 18421129 TI - Dynamical calculation for X-ray 24-beam diffraction in a two-plate crystal cavity of silicon. AB - The X-ray back diffraction of (1240) in a monolithic two-plate silicon cavity occurs at photon energy 14.4388 keV, at which 24 beams are simultaneously excited. Based on the dynamical theory of X-ray diffraction, a theoretical approach has been developed for solving the fundamental equation of dynamical theory to investigate this back diffraction and the interference patterns generated by the Fabry-Perot-type resonance that produces intensity undulation in both transmitted and back-reflected beams. The section of dispersion surface and its associated linear absorption coefficients, wavefield intensities and excitation of mode are calculated. The calculated intensity distribution of the transmitted beam is in a good agreement with the observed one. Details about the interaction between the multiply diffracted X-rays and cavity resonant photons are also reported. Procedures of computer programming are also provided. PMID- 18421130 TI - Foundations of residual-density analysis. AB - New and concise descriptors of the residual density are presented, namely the gross residual electrons, the net residual electrons and the fractal dimension distribution. These descriptors indicate how much residual density is present and in what way it is distributed, i.e. the extent to which the distribution is featureless. The amount of residual density present accounts for noise in the experimental data as well as for modeling inadequacies. Therefore, the minimization of the gross residual electrons during refinement serves as a quality criterion. In the case where only Gaussian noise is present in the residual density, the fractal distribution is parabolic in shape. Deviations from this shape therefore serve as an indicator for systematic errors. The new measures have been applied to simulated and experimental data in order to study the effects of noise, model inadequacies and truncation in the experimental resolution. These measures, although designed and examined with particular regard to applications of space residual density, are very general and can in principle also be applied to space and momentum residual densities in a one-, two-, three- or higher-dimensional Euclidean space. PMID- 18421131 TI - Tables of crystallographic properties of magnetic space groups. AB - Tables of crystallographic properties of the reduced magnetic superfamilies of space groups, i.e. the 7 one-dimensional, 80 two-dimensional and 1651 three dimensional group types, commonly referred to as magnetic space groups, are presented. The content and format are similar to that of non-magnetic space groups and subperiodic groups given in International Tables for Crystallography. Additional content for each representative group of each magnetic space-group type includes a diagram of general positions with corresponding general magnetic moments, Seitz notation used as a second notation for symmetry operations, and general and special positions listed with the components of the corresponding magnetic moments allowed by symmetry. PMID- 18421132 TI - Three-periodic nets and tilings: regular and related infinite polyhedra. AB - The six infinite regular (flag-transitive) polyhedra with finite faces of Grunbaum and Dress are described as tilings of the P and D periodic minimal surfaces. The three polyhedra formed by analogous tiling of the G surface are also described. The nets of these polyhedra are identified. It is shown how these polyhedra, and the nets they carry, could be found by mining the EPINET database of structures. The nets of regular three-periodic polyhedra with infinite helical or zigzag faces are also identified. PMID- 18421133 TI - A new simple tiling, with unusual properties, by a polyhedron with 14 faces. AB - A monotypic simple tiling by a 14-face polyhedron that does not admit an isohedral tiling is described. The tiling is triclinic and contains four distinct, but combinatorially equivalent, kinds of tile. PMID- 18421137 TI - Protein disulfide bond generation in Escherichia coli DsbB-DsbA. AB - Protein disulfide bond formation is catalyzed by a series of Dsb enzymes present in the periplasm of Escherichia coli. The crystal structure of the DsbB-DsbA ubiquinone ternary complex provided important insights into mechanisms of the de novo disulfide bond generation cooperated by DsbB and ubiquinone and of the disulfide bond shuttle from DsbB to DsbA. The structural basis for prevention of the crosstalk between the DsbA-DsbB oxidative and the DsbC-DsbD reductive pathways has also been proposed. PMID- 18421138 TI - Ultrahigh-resolution crystallography and related electron density and electrostatic properties in proteins. AB - With an increasing number of biological macromolecular crystal structures measured at ultrahigh resolution (1 A or better), it is necessary to extend to large systems the experimental valence electron density modelling that is applied to small molecules. A database of average multipole populations has been built, describing the electron density of chemical groups in all 20 amino acids found in proteins. It allows calculation of atomic aspherical scattering factors, which are the starting point for refinement of the protein electron density, using the MoPro software. It is shown that the use of non-spherical scattering factors has a major impact on crystallographic statistics and results in a more accurate crystal structure, notably in terms of thermal displacement parameters and bond distances involving H atoms. It is also possible to obtain a realistic valence electron density model, which is used in the calculation of the electrostatic potential and energetic properties of proteins. PMID- 18421139 TI - Key steps in the structure-based optimization of the hepatitis C virus NS3/4A protease inhibitor SCH503034. AB - The structures of both native and S139A holo-HCV NS3/4A protease domain were solved to high resolution. Subsequently, structures were determined for a series of ketoamide inhibitors in complex with the protease. The changes in the inhibitor potency were correlated with changes in the buried surface area upon binding the inhibitor to the active site. The largest contributions to the binding energy arise from the hydrophobic interactions of the P1 and P2 groups as they bind to the S1 and S2 pockets. This correlation of the changes in potency with increased buried surface area contributed directly to the design of a potent tripeptide inhibitor of the HCV NS3/4A protease, which is currently in clinical trials. PMID- 18421140 TI - Structural studies on Helicobacter pyloriATP-dependent protease, FtsH. AB - The ATP-dependent protease, FtsH, degrades misassembled membrane proteins for quality control like SecY, subunit a of FoF1-ATPase, and YccA, and digests short lived soluble proteins in order to control their cellular regulation, including sigma32, LpxC and lambdacII. The FtsH protein has an N-terminal transmembrane segment and a large cytosolic region that consists of two domains, an ATPase and a protease domain. To provide a structural basis for the nucleotide-dependent domain motions and a better understanding of substrate translocation, the crystal structures of the Helicobacter pylori (Hp) FtsH ATPase domain in the nucleotide free state and complexed with ADP, were determined. Two different structures of HpFtsH ATPase were observed, with the nucleotide-free state in an asymmetric unit, and these structures reveal the new forms and show other conformational differences between the nucleotide-free and ADP-bound state compared with previous structures. In particular, one HpFtsH Apo structure has a considerable rotation difference compared with the HpFtsH ADP complex, and this large conformational change reveals that FtsH may have the mechanical force needed for substrate translocation. PMID- 18421141 TI - Structure of six-transmembrane cation channels revealed by single-particle analysis from electron microscopic images. AB - Six-transmembrane (6-TM) cation channels are plasma membrane-integral components of cellular signaling pathways conserved in almost all species, including animals, plants and some kinds of prokaryotes. These channels selectively permeate cations in response to various signals. In excitable and non-excitable mammalian cells, 6-TM cation channels play fundamental roles, including the generation of action potential and its transmission, the regulation of intracellular ion concentrations, and the activation of signaling cascades by humoral or mechanical pathways. Recently, the structures of three different 6-TM type cation channels have been determined using single-particle analysis from cryo-electron microscopy images: the voltage-sensitive sodium channel, the IP(3) receptor and the TRPC3 channel. The basic structure of the molecules is similar: a bell-like shape comprising a relatively small extracellular (or luminal) domain, a protein-dense transmembrane domain and an expanded cytoplasmic domain. However, in detail, the cytoplasmic architectures are different from one another and are diversely evolved to their specific physiological functions. PMID- 18421142 TI - Protein structures by spallation neutron crystallography. AB - The Protein Crystallography Station at Los Alamos Neutron Science Center is a high-performance beamline that forms the core of a capability for neutron macromolecular structure and function determination. This capability also includes the Macromolecular Neutron Crystallography (MNC) consortium between Los Alamos (LANL) and Lawrence Berkeley National Laboratories for developing computational tools for neutron protein crystallography, a biological deuteration laboratory, the National Stable Isotope Production Facility, and an MNC drug design consortium between LANL and Case Western Reserve University. PMID- 18421143 TI - Solution structures of RseA and its complex with RseB. AB - The bacterial envelope stress response, which is responsible for sensing stress signals in the envelope and for turning on the sigma(E)-dependent transcription, is modulated by the binding of RseB to RseA. In this study, the solution structures of RseA and its complex with RseB were analyzed using circular dichroism and small-angle X-ray scattering. The periplasmic domain of RseA is unstructured and flexible when it is not bound to RseB. However, upon the formation of the stable complex with RseB, RseA induces conformational changes in RseB and, at the same time, RseA becomes more structured. Furthermore, it appears that some other undefined region of RseA, as well as the previously identified minimum region (amino acid 169-186), is also involved in RseB binding. It is thought that these conformational changes are relevant to the proteolytic cleavage of RseA and the modulation of envelope stress response. PMID- 18421144 TI - An optimal exposure strategy for cryoprotected virus crystals with lattice constants greater than 1000 A. AB - Studies of icosahedral virus capsids provide insights into the function of supramolecular machines. Virus capsid crystals have exceptionally large unit cells; as a result, they diffract weakly compared with protein crystals. HK97 is a dsDNA lambda-like bacteriophage whose 13 MDa capsid expands from 550 A to 650 A with large subunit conformational changes during virus maturation. The HK97 penultimate maturation intermediate was crystallized in a tetragonal unit cell that has lattice constants of 1010 A x 1010 A x 730 A. The crystals could be cryoprotected, but diffracted to a modest resolution of 5 A at a bending-magnet beamline. When these crystals were optimally exposed with two orders-of-magnitude more photons from a new insertion-device beamline, data extending to better than 3.8 A resolution were obtained. Here, the strategies to collect and process such data are described. These strategies can be adapted for other crystals with large unit cells and for microcrystals. PMID- 18421145 TI - Fragment approaches in structure-based drug discovery. AB - There has been considerable interest recently in what is known as ;fragment-based lead discovery'. The novel feature of the approach is to begin with small low affinity compounds. The main advantage is that a larger potential chemical diversity can be sampled with fewer compounds, which is particularly important for new target classes. The approach relies on careful design of the fragment library, a method that can detect binding of the fragment to the protein target, determination of the structure of the fragment bound to the target, and the conventional use of structural information to guide compound optimization. In this article the methods are reviewed, and experiences in fragment-based discovery of lead series of compounds against kinases such as PDK1 and ATPases such as Hsp90 are discussed. The examples illustrate some of the key benefits and issues of the approach and also provide anecdotal examples of the patterns seen in selectivity and the binding mode of fragments across different protein targets. PMID- 18421146 TI - Highly bright X-ray generator using heat of fusion with a specially designed rotating anticathode. AB - A new type of rotating anticathode X-ray generator has been developed, in which the electron beam irradiates the inner surface of a U-shaped anticathode (Cu). A high-flux electron beam is focused on the inner surface by optimizing the shape of the bending magnet. The power of the electron beam can be increased to the point at which the irradiated part of the inner surface is melted, because a strong centrifugal force fixes the melted part on the inner surface. When the irradiated part is melted, a large amount of energy is stored as the heat of fusion, resulting in emission of X-rays 4.3 times more brilliant than can be attained by a conventional rotating anticathode. Oscillating translation of the irradiated position on the inner surface during use is expected to be very advantageous for extending the target life. A carbon film coating on the inner surface is considered to suppress evaporation of the target metal and will be an important technique in further realization of highly bright X-ray generation. PMID- 18421147 TI - High-resolution X-ray microdiffraction analysis of natural teeth. AB - The main component of natural teeth was determined many years ago as calcium phosphate, mostly in the form of hydroxyapatite with different crystallites. In the past, the method used in tooth crystal investigation has been mainly powder X ray diffraction analysis, but this method has its drawbacks, i.e. the destruction of the natural tooth structure and the difficulty in examining the preferred orientation in different layers of the tooth. During the last century, microzone X-ray diffraction on the tooth surface was carried out, but, as the technology was less sophisticated, the results obtained were not very detailed. The newly developed microdiffraction equipment permits analysis of the microzone of teeth in situ. To test this new microdiffraction equipment, microdiffraction analysis of one natural healthy deciduous molar tooth and one carious deciduous molar tooth has been performed, using a Bruker D8 instrument. Phase analysis of the two teeth was performed; the crystal size at six test points in the natural healthy tooth was calculated by reflection (211), and the crystal preferred orientation of reflection (300) and reflection (002) at six test points in the natural healthy tooth were compared. The results showed that the tooth was a kind of biological mixed crystal composed of several crystal phases, the main crystal phase being hydroxyapatite. The crystal size grew larger going from the dentin to the enamel. The crystal preferred orientation mainly existed in the enamel, especially in the reflection (002). From our experiment, layer orientation and continuous crystal variations in teeth could be conveniently studied using fast online measurements by high-resolution X-ray microdiffraction equipment. PMID- 18421148 TI - Interaction of HIV-1 aspartic protease with its inhibitor, by molecular dynamics and ab initio fragment molecular orbital method. AB - For the three complex crystal structures of HIV-1 aspartic protease (an enzyme of AIDS) with its inhibitor in the Protein Data Bank, molecular dynamics of the generalized Born surface area and the ab initio fragment molecular orbital of an ABINIT-MP calculation was performed to obtain the binding free energy, the molecular orbital energy, the interaction energy of residues with an inhibitor and the charge transfer at the active site. The inhibitors are five symmetric cyclic ureas, of which three were modelled, and an asymmetric dipeptide. The interaction energy of the inhibitor at the active sites of aspartic acid is as great as 50 kcal mol(-1), coinciding with a tetrahedral transition state. For the inhibitor with a higher affinity, charge was transferred to the inhibitor from the active site. The difference in symmetry of the inhibitor was not evident. Binding free energy corresponds to the experimental value of the binding constant, while molecular orbital energy does not always, which is considered to be an entropy effect. PMID- 18421149 TI - Trimeric structure and conformational equilibrium of M-ficolin fibrinogen-like domain. AB - Ficolins are pathogen-recognition molecules in innate immune systems. The crystal structure of the human M-ficolin recognition domain (FD1) has been determined at 1.9 A resolution, and compared with that of the human fibrinogen gamma fragment, tachylectin-5A, L-ficolin and H-ficolin. The overall structure of FD1 is similar to that of the other proteins, although the peptide bond between Asp282 and Cys283, which is in a predicted ligand-binding site, is a normal trans bond, unlike the cases of the other proteins. Analysis of the pH-dependent ligand binding activity of FD1 in solution suggested that a conformational equilibrium between active and non-active forms in the ligand-binding region, involving cis trans isomerization of the Asp282-Cys283 peptide bond, contributes to the discrimination between self and non-self, and that the pK(a) values of His284 are 6.1 and 6.3 in the active and non-active forms, respectively. PMID- 18421150 TI - A degradation signal recognition in prokaryotes. AB - The degradation of ssrA-tagged substrates in prokaryotes is conducted by a subset of ATP-dependent proteases, including ClpXP complex. More than 630 sequences of ssrA have been identified from 514 species, and are conserved in a wide range of prokaryotes. SspB protein markedly stimulates the degradation of these ssrA tagged substrates by the ClpXP proteolytic machine. The dimeric SspB protein is composed of a compact ssrA-binding domain, which has a dimerization surface and a flexible C-terminal tail with a ClpX-binding motif at its very end. Since SspB is an adaptor protein for the ClpXP complex, designed mutagenesis, fluorescence spectroscopy, biochemistry and X-ray crystallography have been used to investigate the mechanism of delivery of ssrA-tagged proteins. In this paper the structural basis of ssrA-tag recognition by ClpX and SspB, as well as SspB-tail recognition by ZBD, is described. PMID- 18421151 TI - Deduced catalytic mechanism of D-amino acid amidase from Ochrobactrum anthropi SV3. AB - D-Amino acid amidase (DAA) from Ochrobactrum anthropi SV3 catalyzes D stereospecific hydrolysis of amino acid amides. DAA has attracted attention as a catalyst for the stereospecific production of D-amino acids, although the mechanism that drives the reaction has not been clear. Previously, the structure of DAA was classified into two types, a substrate-bound state with an ordered Omega loop, and a ground state with a disordered Omega loop. Because the binding of the substrate facilitates ordering, this transition was regarded to be induced fit motion. The angles and distances of hydrogen bonds at Tyr149 Oeta, Ser60 Ogamma and Lys63 Nzeta revealed that Tyr149 Oeta donates an H atom to a water molecule in the substrate-bound state, and that Tyr149 Oeta donates an H atom to Ser60 Ogamma or Lys63 Nzeta in the ground state. Taking into consideration the locations of the H atoms of Tyr149 Oeta, Ser60 Ogamma and Lys63 Nzeta, a catalytic mechanism of DAA activity is presented, wherein a shift of an H atom at Tyr149 Oeta in the substrate-bound versus the ground state plays a significant role in the reaction. This mechanism explains well why acylation proceeds and deacylation does not proceed in the substrate-bound state. PMID- 18421152 TI - Novel dimer structure of a membrane-bound protease with a catalytic Ser-Lys dyad and its linkage to stomatin. AB - Membrane-bound proteases are involved in various regulatory functions. A previous report indicates that the N-terminal region of PH1510 (1510-N) from the hyperthermophilic archaeon Pyrococcus horikoshii is a serine protease with a catalytic Ser-Lys dyad (Ser97 and Lys138), and specifically cleaves the C terminal hydrophobic region of the p-stomatin PH1511. According to the crystal structure of the wild-type 1510-N in dimeric form, the active site around Ser97 is in a hydrophobic environment suitable for the hydrophobic substrates. This article reports the crystal structure of the K138A mutant of 1510-N at 2.3 A resolution. The determined structure contains one molecule per asymmetric unit, but 1510-N is active in dimeric form. Two possible sets of dimer were found from the symmetry-related molecules. One dimer is almost the same as the wild-type 1510-N. Another dimer is probably in an inactive form. The L2 loop, which is disordered in the wild-type structure, is significantly kinked at around A-138 in the K138A mutant. Thus Lys138 probably has an important role on the conformation of L2. PMID- 18421153 TI - Performance of an electron gun for a high-brightness X-ray generator. AB - A prototype thermionic electron gun for a high-brightness X-ray generator has been developed. Its extraction voltage and design current are 60 kV and 100 mA (DC), respectively. The X-ray generator aims towards a maximum brilliance of 60 kW mm(-2). The beam sizes at the rotating anticathode must therefore be within 1.0 mm x 0.1 mm and a small beam emittance is required. The fabricated electron gun optimizes an aperture grid and a Whenelt electrode. The performance of the prototype electron gun measured using pulsed-beam tests is as follows: maximum beam current, 85.7 mA; beam focus size at the rotating anticathode, 0.79 mm x 0.13 mm. In DC beam tests, FWHM beam sizes were measured to be 0.65 mm x 0.08 mm at the rotating anticathode with a beam current of 45 mA. The beam current recently reached approximately 60 mA with some thermal problems. PMID- 18421154 TI - Crystal twinning of human MD-2 recognizing endotoxin cores of lipopolysaccharide. AB - Twinning of crystals causes overlapping of two or more reciprocal lattice points, and hence structure amplitudes for a single crystalline domain are hardly obtained from X-ray diffraction intensities. MD-2 protein forms a stable complex with Toll-like receptor 4 and recognizes bacterial lipopolysaccharide (LPS). Excessive immune responses activated by LPS cause septic shocks. Saccharide trimmed human MD-2 crystallizes in the tetragonal form with apparent Laue symmetry of 4/mmm, and diffraction intensities from these crystals indicate crystal twinning. The crystal consists of two different domains, A and B. The c(A) axis of domain A coincides with the c(B) axis of domain B with a smaller lattice, and the a(A) axis corresponds to the (a(B) + b(B)) axis. This twinning severely imposes difficulty in structure determination. Through optimization of cryoprotectant, domain A was thoroughly transformed into domain B. The crystal containing only domain B is in space group P4(1)2(1)2 with one MD-2 molecule in the asymmetric unit. The structure of this form of MD-2 as well as its complex with antiendotoxic lipid IVa was successfully determined using the multiple isomorphous replacement method. PMID- 18421155 TI - Crystallization of the Atg12-Atg5 conjugate bound to Atg16 by the free-interface diffusion method. AB - Autophagy mediates the bulk degradation of cytoplasmic components in lysosomes/vacuoles. Five autophagy-related (Atg) proteins are involved in a ubiquitin-like protein conjugation system. Atg12 is conjugated to its sole target, Atg5, by two enzymes, Atg7 and Atg10. The Atg12-Atg5 conjugates form a multimeric complex with Atg16. Formation of the Atg12-Atg5-Atg16 ternary complex is crucial for the functions of these proteins on autophagy. Here, the expression, purification and crystallization of the Atg12-Atg5 conjugate bound to the N-terminal region of Atg16 (Atg16N) are reported. The Atg12-Atg5 conjugates were formed by co-expressing Atg5, Atg7, Atg10 and Atg12 in Eschericia coli. The Atg12-Atg5-Atg16N ternary complex was formed by mixing purified Atg12-Atg5 conjugates and Atg16N, and was further purified by gel-filtration chromatography. Crystallization screening was performed by the free-interface diffusion method. Using obtained microcrystals as seeds, large crystals for diffraction data collection were obtained by the sitting-drop vapour-diffusion method. The crystal contained one ternary complex per asymmetric unit, and diffracted to 2.6 A resolution. PMID- 18421156 TI - Protein crystallization in a 100 nl solution with new stirring equipment. AB - To investigate quantitatively the effects of stirring on protein crystallization, a new stirring system which can agitate a protein solution, approximately 100 nl, by providing Hagen-Poiseuille flow has been successfully developed. In addition, this new stirring system provides flow with a well defined pattern and velocity. Using this system, hen egg-white lysozyme was crystallized in 100-200 nl solutions while being stirred. The optimum stirring conditions for lysozyme crystals have been explored by evaluating the Reynolds (Re) number and the crystals obtained. Intermittent flow, as well as a low Re number, was found to contribute significantly to the growth of a smaller number of larger crystals. PMID- 18421157 TI - Structural insights into the similar modes of Nrf2 transcription factor recognition by the cytoplasmic repressor Keap1. AB - The cytoplasmic repressor Keap1 regulates the function of transcription factor Nrf2 which plays critical roles in oxidative and xenobiotic stresses. The Neh2 domain of Nrf2 interacts with Keap1 at the bottom region of the Kelch/beta propeller domain which is formed by double-glycine repeat and C-terminal region domains (Keap1-DC). The structure of Keap1-DC complexed with an Nrf2 peptide containing a conserved DLG motif has been determined at 1.9 A resolution. The Keap1-bound DLG peptide possesses a hairpin conformation, and it binds to the Keap1 protein at the bottom region of the beta-propeller domain. The intermolecular interaction occurs through their complementary electrostatic interactions. Comparison of the present structure with the recently reported Keap1-DC complex structure suggests that the DLG and ETGE motifs of Neh2 in Nrf2 bind to Keap1 in a similar manner but with different binding potencies. PMID- 18421158 TI - Hydrogen bonds of DsrD protein revealed by neutron crystallography. AB - The features of hydrogen bonds in DsrD protein from sulfate-reducing bacteria have been investigated by neutron protein crystallography. The function of DsrD has not yet been elucidated clearly, but its X-ray crystal structure revealed that it comprises a winged-helix motif and shows the highest structural homology to the DNA-binding proteins. Since any neutron structure of a DNA recognition protein has not yet been obtained, here detailed information on the hydrogen bonds in the winged-helix-motif protein is given and the following features found. (i) The number of hydrogen bonds per amino acid of DsrD is relatively fewer than for other proteins for which neutron structures were determined previously. (ii) Hydrogen bonds are localized between main-chain and main-chain atoms; there are few hydrogen bonds between main-chain and side-chain atoms and between side-chain and side-chain atoms. (iii) Hydrogen bonds inducted by protonation of specific amino acid residues (Glu50) seem to play an essential role in the dimerization of DsrD. The former two points are related to the function of the DNA-binding protein; the three-dimensional structure was mainly constructed by hydrogen bonds in main chains, while the side chains appeared to be used for another role. The latter point would be expected to contribute to the crystal growth of DsrD. PMID- 18421159 TI - Development of an X-ray HARP-FEA detector system for high-throughput protein crystallography. AB - A new detector system for protein crystallography is now being developed based on an X-ray HARP-FEA (high-gain avalanche rushing amorphous photoconductor-field emitter array), which consists of an amorphous selenium membrane and a matrix field emitter array. The combination of the membrane avalanche effect with a single driven FEA has several advantages over currently available area detectors, including higher sensitivity, higher spatial resolution and a higher frame rate. Preliminary evaluation of the detector has been carried out and its effectiveness has been confirmed. Next, diffraction images were measured with continuous rotation of a protein crystal, and the images were compared with those measured by the existing CCD detector; the system successfully obtained high-spatial resolution images. Using shutterless measurement, the total measurement time can be reduced significantly, making the method appropriate for high-throughput protein crystallography. The X-ray HARP-FEA detector is an attractive candidate for the next generation of X-ray area detectors. PMID- 18421160 TI - Mutagenesis of the crystal contact of acidic fibroblast growth factor. AB - An attempt has been made to improve a crystal contact of human acidic fibroblast growth factor (haFGF; 140 amino acids) to control the crystal growth, because haFGF crystallizes only as a thin-plate form, yielding crystals suitable for X ray but not neutron diffraction. X-ray crystal analysis of haFGF showed that the Glu81 side chain, located at a crystal contact between haFGF molecules, is in close proximity with an identical residue related by crystallographic symmetry, suggesting that charge repulsion may disrupt suitable crystal-packing interactions. To investigate whether the Glu residue affects the crystal-packing interactions, haFGF mutants in which Glu81 was replaced by Ala, Val, Leu, Ser and Thr were constructed. Although crystals of the Ala and Leu mutants were grown as a thin-plate form by the same precipitant (formate) as the wild type, crystals of the Ser and Thr mutants were grown with increased thickness, yielding a larger overall crystal volume. X-ray structural analysis of the Ser mutant determined at 1.35 A resolution revealed that the hydroxy groups of Ser are linked by hydrogen bonds mediated by the formate used as a precipitant. This approach to engineering crystal contacts may contribute to the development of large protein crystals for neutron crystallography. PMID- 18421161 TI - Mail-in data collection at SPring-8 protein crystallography beamlines. AB - A mail-in data collection system makes it possible for beamline users to collect diffraction data without visiting a synchrotron facility. In the mail-in data collection system at SPring-8, users pack crystals into sample trays and send the trays to SPring-8 via a courier service as the first step. Next, the user specifies measurement conditions and checks the diffraction images via the Internet. The user can also collect diffraction data using an automated sample changer robot and beamline control software. For distant users there is a newly developed data management system, D-Cha. D-Cha provides a graphical user interface that enables the user to specify the experimental conditions for samples and to check and download the diffraction images using a web browser. This system is now in routine operation and is contributing to high-throughput beamline operation. PMID- 18421162 TI - X-ray beam stabilization at BL-17A, the protein microcrystallography beamline of the Photon Factory. AB - BL-17A is a new structural biology beamline at the Photon Factory, Japan. The high-brilliance beam, derived from the new short-gap undulator (SGU#17), allows for unique protein crystallographic experiments such as data collection from microcrystals and structural determination using softer X-rays. However, microcrystal experiments require robust beam stability during data collection and minor fluctuations could not be ignored. Initially, significant beam instability was observed at BL-17A. The causes of the beam instability were investigated and its various sources identified. Subsequently, several effective countermeasures have been implemented, and the fluctuation of the beam intensity successfully suppressed to within 1%. Here the instability reduction techniques used at BL-17A are presented. PMID- 18421163 TI - Implementation of remote monitoring and diffraction evaluation systems at the Photon Factory macromolecular crystallography beamlines. AB - Owing to recent advances in high-throughput technology in macromolecular crystallography beamlines, such as high-brilliant X-ray sources, high-speed readout detectors and robotics, the number of samples that can be examined in a single visit to the beamline has increased dramatically. In order to make these experiments more efficient, two functions, remote monitoring and diffraction image evaluation, have been implemented in the macromolecular crystallography beamlines at the Photon Factory (PF). Remote monitoring allows scientists to participate in the experiment by watching from their laboratories, without having to come to the beamline. Diffraction image evaluation makes experiments easier, especially when using the sample exchange robot. To implement these two functions, two independent clients have been developed that work specifically for remote monitoring and diffraction image evaluation. In the macromolecular crystallography beamlines at PF, beamline control is performed using STARS (simple transmission and retrieval system). The system adopts a client-server style in which client programs communicate with each other through a server process using the STARS protocol. This is an advantage of the extension of the system; implementation of these new functions required few modifications of the existing system. PMID- 18421164 TI - High-throughput operation of sample-exchange robots with double tongs at the Photon Factory beamlines. AB - Sample-exchange robots that can exchange cryo-pins bearing protein crystals out of experimental hutches according to user instructions have been developed. The robots were designed based on the SAM (Stanford Synchrotron Research Laboratory automated mounting) system. In order to reduce the time required for the sample exchange, the single tongs of the SAM system were modified and a double-tongs system that can hold two cryo-pins at the same time was developed. Robots with double tongs can move to the goniometer head holding the next cryo-pin with one set of tongs, dismount the experimented cryo-pin with the other set, and then mount the next pin onto the goniometer head without leaving the diffractometer area. Two different types of tongs have been installed: single tongs at beamlines BL-5A and AR-NW12A, and a double-tongs system at beamline BL-17A of the Photon Factory. The same graphical user interface software for operation of the sample exchange robots is used at all beamlines, however, so that users do not need to consider differences between the systems. In a trial, the robot with double tongs could exchange samples within 10 s. PMID- 18421165 TI - X-ray crystallographic and biochemical characterizations of a mutant photosystem II complex from Thermosynechococcus vulcanus with the psbTc gene inactivated by an insertion mutation. AB - The crystal structure of a photosystem II (PSII) dimer from Thermosynechococcus vulcanus with its psbTc gene inactivated by insertion mutation of an antibiotic cassette in a site in the C-terminal region was analyzed at 3.8 A resolution. In the crystal structure of the mutant PSII, the transmembrane helix of PsbTc remains, whereas the C-terminal loop of PsbTc has disappeared. In addition, the PsbM subunit, which seemed to be lost in a PsbTc-deletion mutant PSII of T. elongatus, is still present. The deletion of the C-terminal loop of PsbTc in the mutant PSII was verified by mass spectrometry. Thus, the insertion mutation of psbTc eliminated only the C-terminal loop of this subunit. Nevertheless, some features of the mutant PSII, namely a destabilization of the dimeric form and a slight decrease of the oxygen-evolving activity, were observed in the mutant, indicating that the C-terminal loop of PsbTc functions to maintain the stability of the PSII dimer and the activity of oxygen evolution. PMID- 18421166 TI - Complex of human neutrophil elastase with 1/2SLPI. AB - SLPI (secretory leukocyte protease inhibitor) is a 107-residue non-glycosylated protease inhibitor, which inhibits a wide range of serine proteases, trypsin, chymotrypsin, neutrophil elastase, chymase and cathepsin G. X-ray crystallographic analyses have shown that SLPI comprises two separate domains of similar architecture [Grutter, Fendrich, Huber & Bode (1988), EMBO J. 7, 345-351] and the C-terminal domain interacts with bovine alpha-chymotrypsin. In order to understand SLPI's multiple functions against various serine proteases, the complex HNE (human neutrophil elastase) has been co-crystallized with 1/2SLPI (recombinant C-terminal domain of SLPI; Arg58-Ala107), which has a biological activity similar to full SLPI. The 1/2SLPI and HNE complex structure was solved at 1.7 A resolution, and compared with the interaction mechanism of elafin, which is a specific inhibitor of elastase. It was found that P1 Leu72i and six hydrogen bonds between the main chains in the primary contact region have sufficient ability to inhibit HNE and PPE (porcine pancreatic elastase), and P5 Tyr68i is important in increasing the selectivity of 1/2SLPI against HNE. The mechanisms of the functions of SLPI are relatively unknown, but the current study could help understand the selectivity of SLPI against HNE and PPE. PMID- 18421167 TI - Crystallization and evaluation of hen egg-white lysozyme crystals for protein pH titration in the crystalline state. AB - To observe the ionized status of the amino acid residues in proteins at different pH (protein pH titration in the crystalline state) by neutron diffraction, hen egg-white lysozyme was crystallized over a wide pH range (2.5-8.0). Crystallization phase diagrams at pH 2.5, 6.0 and 7.5 were determined. At pH < 4.5 the border between the metastable region and the nucleation region shifted to the left (lower precipitant concentration) in the phase diagram, and at pH > 4.5 the border shifted to the right (higher precipitant concentration). The qualities of these crystals were characterized using the Wilson plot method. The qualities of all crystals at different pH were more or less equivalent (B-factor values within 25-40). It is expected that neutron diffraction analysis of these crystals of different pH provides equivalent data in quality for discussions of protein pH titration in the crystalline state of hen egg-white lysozyme. PMID- 18421168 TI - Effect of amino acids and amino acid derivatives on crystallization of hemoglobin and ribonuclease A. AB - Determination of the appropriate conditions for protein crystallization remains a highly empirical process. Preventing protein aggregation is necessary for the formation of single crystals under aggregation-prone solution conditions. Because many amino acids and amino acid derivatives offer a unique combination of solubility and stabilizing properties, they open new avenues into the field of protein aggregation research. The use of amino acids and amino acid derivatives can potentially influence processes such as heat treatment and refolding reactions. The effect of the addition of several amino acids, such as lysine, and several amino acid derivatives, such as glycine ethyl ester and glycine amide, on the crystallization of equine hemoglobin and bovine pancreatic ribonuclease A has been examined. The addition of these amino acids and amino acid derivatives expanded the range of precipitant concentration in which crystals formed without aggregation. The addition of such additives appears to promote the crystallization of proteins. PMID- 18421169 TI - Proceedings of the 2nd International Symposium on Diffraction Structural Biology. September 10-13, 2007. Tokyo, Japan. PMID- 18421170 TI - Developmental abnormalities of the thymus in hea/hea mutant mice. AB - This study found that the thymus of hea/hea mutant mice (hea mice) became atrophic in early phase of life and that the differentiation of CD4-CD8- (Double Negative; DN) into CD4+CD8+ (Double Positive; DP) cells during the development of T cells in the thymus was abnormal. The thymus development of hea mice was different from that of normal littermates. After 6 days of age, the numbers of thymocytes in hea mice decreased. The total numbers of DP cells in the thymus of hea mice reached the maximum at 6-9 days of age and then decreased after 10 days. The total numbers of DN cells in the thymus were almost constant in hea mice and normal littermates. These results indicate abnormalities in the process of differentiation from DN to DP cells in the thymus of hea mice. Flow cytometoric analysis indicated the presence of a large number of apoptotic and necrotic cells in the thymus of 13-15-day-old hea mice. However, there were no significant differences in the amount of mRNAs of Fas, Fas ligand and IL-7 between hea mice and normal littermates. Splenocytes from hea mice produced the same amount of cytokine mRNAs as normal littermate mice and the hea mutation (Ttc7(fsn-hea)) did not affect serum levels of IgM immunoglobulin. However, activated T cells from hea mice showed more secretion of cytokines derived from Th2 cells than from Th1 cells, so they might be affected by abnormalities of the immune system. PMID- 18421171 TI - Inhomogeneity of fecal flora in separately reared laboratory mice, as detected by denaturing gradient gel electrophoresis (DGGE). AB - Laboratory mice were divided into 2 groups and introduced to different rooms immediately after being transferred from a mouse farm. Polymerase chain reaction followed by denaturing gradient gel electrophoresis were performed on the V6-V8 regions of bacterial 16S rDNA obtained from fecal samples at 0, 1, 2, 3, 4 and 8 weeks after the introduction. Binary data were obtained from banding patterns, and Euclidean distances for each week were calculated and analyzed by cluster analysis and non-metric multidimensional scaling. Euclidean distances were significantly higher at weeks 1 and 2 than at week 0 in both groups, although the distances between the 2 groups were significantly higher after week 1 than week 0. The distances between the 2 groups were significantly higher than those within each group at weeks 4 and 8. Mice in the 2 groups formed clusters at weeks 2 and 3 respectively, and mice were divided into 2 clusters by their respective groups at weeks 4 and 8. Mice in the 2 groups were distributed on opposite sides of the origin on the 2-dimensional plane after week 2. These results suggest that mouse fecal flora changed characteristically, according to the local environment after introduction. PMID- 18421172 TI - Clinical and histopathological evaluation of MDP/collagen induced arthritis rat model (MCIA) after treatment with Urtica dioica, plantago major and Hypericum perforatum L herbal mixture. AB - This study was done to assess the effects of Urtica dioica, Plantago major and Hypericum perforatum L herbal mixture in the MCIA rat model. In addition, a new pathological and clinical arthritis lesion assessment was developed. Sprague Dawley (SD) rats were immunized with bovine type II collagen and muramyl dipeptide (MDP). Commercial herbal extracts were administered daily to the rats after the immunization for the course of experiment (90 days). Rats were boosted with a second collagen-MDP emulsion 60 days after the first immunization. Paws were daily evaluated macroscopically for redness, swelling, distortion, or ankylosis of the joints. On the day of sacrifice, rat paws were assessed for histopathologic changes. Herbal mixture administration decreased the clinical lesion manifestation in the MCIA rat model and led to development of similar or slightly more severe histopathological lesions compared to rats that did not receive the treatment. The clinical arthritis signs appeared as early as 13 days after the first MDP/collagen injection and with peak incidence at 20 days post immunization. Histopathologically, animals showed changes ranging from mild to very severe. Administration of the herbal mixture used in this study had a clinical therapeutic effect on the course of the clinical manifestations in the MCIA model, but the herbal treatment had no such effect on the histopathological lesion development and even led to slightly more severe lesions. Rats in the MCIA model developed prominent clinical and histopathological changes that were comparable to rheumatoid arthritis (RA) lesions in humans. PMID- 18421173 TI - Diabetes-associated complications in Spontaneously Diabetic Torii fatty rats. AB - The Spontaneously Diabetic Torii (SDT) fatty rat, established by introducing the fa allele of the Zucker fatty rat into the SDT rat genome, is a new model of obese type 2 diabetes. The SDT-fa/fa (SDT fatty) rat shows overt obesity, and hyperglycemia and hyperlipidemia are observed at a young age as compared with the SDT-+/+ (SDT normal) rat. However, the features of the diabetic complications in the SDT fatty rat have not been reported. In the present study, the incidence and the progression of diabetic complications in the SDT fatty rat were examined, and compared with those of the SDT normal rat. Renal function parameters, such as blood urea nitrogen, urine volume and urinary protein, increased from 4 weeks of age in the SDT fatty rat, and pathological findings in the renal tubule were observed from 8 weeks. Furthermore, cataract was observed in the SDT fatty rat from 8 weeks of age, and prolongation of peak latencies on electroretinograms was observed at 16 and 24 weeks of age. On the other hand, in the SDT normal rat, renal or ocular changes were observed from 24 weeks of age. With early incidence of diabetes mellitus, diabetes-associated complications in the SDT fatty rat were seen at younger ages than those in the SDT normal rat. In conclusion, the SDT fatty rat is expected to be a useful model for the analysis of diabetic complications and the evaluation of drugs related to metabolic diseases. PMID- 18421174 TI - Developmental responses of 2-cell embryos to oxygen tension and bovine serum albumin in Wistar rats. AB - To improve rat embryo culture conditions, responses of Wistar 2-cell embryos from 2 breeders to oxygen tension (5 vs 20%) and bovine serum albumin (BSA) (0 vs 3 mg/ml) were examined using rat 1-cell embryo culture medium (mR1ECM). Supplementation of 3 mg/ml BSA significantly stimulated and accelerated development to the blastocyst and expanded blastocyst stages during 72 and 96 h culture, while reduced oxygen tension stimulated cell division. Fetus development after transfer of blastocysts obtained from 72 h culture under 5% O2 with BSA was significantly higher than those cultured under atmospheric oxygen without BSA. However, the nuclear numbers of in vitro cultured blastocysts and fetus development after embryo transfer were still significantly lower than in vivo developed blastocysts, indicating the current culture condition is still suboptimal. PMID- 18421175 TI - Genetic linkage map of the house musk shrew, Suncus murinus, constructed with PCR based and RFLP markers. AB - A genetic linkage map for Suncus murinus was previously constructed with 11 marker loci. In this study, we developed 172 new microsatellite and three RFLP markers, and re-constructed a new framework map by combining all markers. The new map comprises 42 markers that are distributed into 12 linkage groups, two of which are assigned to chromosomes, and spans 403.5 cM with an average inter marker distance of 13.5 cM. PMID- 18421176 TI - OLETF allele of hyperglycemic QTL Nidd3/of is dominant. AB - The OLETF rat is a well-established model for the study of type 2 diabetes associated with obesity and has been shown to possess multiple hyperglycemic alleles in its genome. Here we focused on and carefully characterized one of the previously reported congenic strains, F.O-Nidd3/of that carries the OLETF allele of the Nidd3/of locus (also known as Niddm21 in the Rat Genome Database) in the normoglycemic F344 genetic background. A prominent finding was that the F1 progeny between the congenic and the F344 stain, whose genotype is heterozygote at the Nidd3/of locus, showed mild hyperglycemia equal to the parental congenic rat, suggesting that the OLETF allele is dominant. To our knowledge, this is the first study in which a diabetic QTL has been directly demonstrated to be dominant by using congenic strains. PMID- 18421177 TI - Clinical pathology data from cynomolgus monkeys from China in which diarrhea was observed during quarantine. AB - Shin Nippon Biomedical Laboratories, Ltd. (SNBL) imported and quarantined 3,148 cynomolgus monkeys (aged 2.5 to 6.5 years) from China in 2002. The hematology and blood biochemistry data obtained from these monkeys on Day 32 of quarantine were analyzed separately by sex [2,890 animals in which no abnormalities were observed during the 35-day quarantine period (normal group), and 258 animals which exhibited diarrhea 1 to 12 times (diarrhea group)]. The values obtained for all parameters were within the normal range (mean +/- SD), and no significant abnormalities were noted in either sex. The clinical pathology data from 11 animals (6 males and 5 females) exhibiting diarrhea repeatedly (10 to 12 times) were statistically analyzed, and significant differences were noted in PLT and ALP in both sexes. The PLT values of these animals were within the normal group mean +/- 2 SD, and were considered within the normal range. A significant difference was noted in some individual ALP values (males: Nos. 2 and 3, females: Nos. 1, 3, and 4). The clinical pathology data obtained from the normal group in this study basically correspond to the widely reported results already obtained from healthy cynomolgus monkeys, from which it can be concluded that the cynomolgus monkeys from China were generally healthy and presenting no particular abnormality. The clinical pathology data from the normal group will serve as valuable baseline data for experimenters using cynomolgus monkeys. PMID- 18421178 TI - Direct comparison between ICSI-mediated DNA transfer and pronuclear DNA microinjection for producing transgenic rats. AB - Production efficiency of transgenic rats was compared directly between the routine pronuclear microinjection of exogenous DNA solution (PNMI-Tg method) and the ooplasmic injection of sperm cells exposed to exogenous DNA solution (ICSI-Tg method) using six DNA constructs. The overall production efficiency per treated oocyte in the ICSI-Tg method (mean 1.1%, range 0.2 to 3.1%) was similar to that in the PNMI-Tg method (mean 1.1%, range 0 to 2.4%). An advantage of the ICSI-Tg method in the production of transgenic rats is noted in cases in which a low yield of pronuclear zygotes is an inevitable fate of the rat strain. PMID- 18421179 TI - Pregnancy and parturition of mice latently infected with Pseudorabies virus. AB - In this study, the influence of pregnancy and parturition on mice in a mouse model of latent infection with Pseudorabies virus (PrV) was analyzed. Latently infected (LI) female mice were paired with mature uninfected males. The mating produced progeny without any clinical signs of Aujeszky's Disease. At weaning, both male and female progeny of LI mice showed significantly lower weight than control mice. PrV was not detected from nasal swab specimens of the female parent mice or the trigeminal ganglia of all mice, except 3 of 50 neonatal mice. These findings demonstrate that pregnancy and parturition induce little reactivation of latent PrV, but do affect the mother's body, as indicated by the decreased weight of progeny at weaning. PMID- 18421180 TI - Alteration of histone tail modifications in the Xist locus in wild-type and Tsix mutant male embryonic stem cells during differentiation. AB - The non-coding RNA Xist is indispensable for X chromosome inactivation. Transcriptional control of Xist gene depends on its antisense partner gene Tsix which prevents Xist up-regulation in cis. Previous studies proposed Tsix acts by regulating chromatin structure. Although histone modifications in the Xist locus during differentiation have been described in female embryonic stem (ES) cells, they remain unclear in males. Here we addressed histone modifications in the Xist locus in wild-type and Tsix-mutant male ES cells during differentiation. Their active and repressive modifications were attenuated upon differentiation, while the histone modification profile in males resembled that of females in an undifferentiated condition. These results provide implications in understanding the regulation of Xist gene, as well as other developmentally regulated genes, through chromatin structure. PMID- 18421181 TI - Pathology of spontaneous dermatitis in CBy.ALY-aly mice. AB - CBy.ALY-aly/aly (C-aly) mice develop progressive dermatitis in areas of the face and dorsal neck from around three months of age. Staphylococcus aureus was detected in the skin lesions of C-aly mice. However, even when the mice were raised under S. aureus free conditions, a similar, though less severe, dermatitis was still observed. The mice showed a marked increase in the number of eosinophils in the peripheral blood and skin lesions, with no changes in plasma IgE levels. These findings suggest that the dermatitis of these mice may be an atypical allergic condition with little or no involvement of IgE. C-aly mice may be a useful animal model of chronic dermatitis or pruritus without elevated IgE levels. PMID- 18421182 TI - Chronic kidney disease in Japan. AB - Chronic kidney disease (CKD) is defined as either kidney damage with urine, imaging, and histologic abnormalities, or a low estimated glomerular filtration rate (GFR) for more than 3 months. The GFR is calculated using either the Modification of Diet in Renal Disease (MDRD) Study equation or the Cockcroft Gault formula. CKD is a risk factor for end-stage renal disease (ESRD) and cardiovascular disease. In Japan, the prevalence of ESRD is increasing and is currently more than 2,000 per million population. More than 40% of incident ESRD is due to diabetes mellitus (DM). The prevalence of a low GFR (< 60 ml/min/1.73 m(2)) is estimated to be 20% of the adult population. Studies based on several community-based screening programs suggest that Japan has a higher prevalence of CKD than any other country. Early detection and treatment of CKD are necessary to decrease the incidence of ESRD and cardiovascular disease. PMID- 18421183 TI - Five-year follow-up study of patients with reflux symptoms and reflux esophagitis in annual medical check-up field. AB - AIM: To clarify changes in the prevalence of reflux symptoms and reflux esophagitis over a period of 5 years. SUBJECTS AND METHODS: Five hundred thirty nine study subjects (male: 408, mean age 47.3 years) were assessed using a reflux symptom questionnaire and endoscopic examinations conducted twice, 5 years apart. RESULTS: At enrollment, 88 patients had reflux symptoms and 44 had reflux esophagitis (RE). After 5 years, 58 and 53 patients were diagnosed as having reflux symptoms and RE, respectively. Only 26% of the patients with reflux symptoms at enrollment had the symptoms after 5 years. The presence of reflux symptoms and an increase of BMI were significant risk factors for the presence of reflux symptoms after 5 years. CONCLUSION: Reflux symptoms frequently disappear after 5 years. It is difficult to identify individuals who will still have symptoms after 5 years on the basis of clinical characteristics. PMID- 18421184 TI - Metformin and parameters of physical health. AB - BACKGROUND: The prevalence of excess weight, including overweight and obesity, is increasing with a high cost on health in society. METHODS: Consecutive cases with excess weight, aged between 50 and 70 years and desiring weight loss, were divided into two subgroups according to wishes of patients about whether they prefer medication or just a diet. Metformin at a daily dose of 2,550 mg was given to the medication group. RESULTS: As for the very high prevalences, 84.8% (313/369) of cases at or above the age of 50 years were overweight or obese, 67.2% (248/369) of them had white coat hypertension (WCH) or hypertension (HT), 52.5% (194/369) of them had impaired glucose tolerance (IGT) or diabetes mellitus (DM), and 68.8% (254/369) of them had dyslipidemia. Initially 143 cases with excess weight preferred the diet and 162 of them preferred the metformin therapy. But 42 cases (25.9%) stopped the drug because of excessive anorexia. At the end of the six-month period, there were highly significant differences between the two groups according to prevalences of resolved WCH, hyperbetalipoproteinemia, hypertriglyceridemia, dyslipidemia, overweight, and obesity and a decreased fasting plasma glucose below 110 mg/dL (p<0.001 for all). CONCLUSION: Due to the very high prevalences of excess weight and probably many associated disorders with the excess weight, including IGT or DM, WCH or HT, and dyslipidemia, above the age of 50 years, and the detected significant benefits of metformin on all of the above parameters, metformin treatment should be initiated in patients with excess weight in their fifties. PMID- 18421185 TI - Plasma gamma-glutamyl transferase activity predicts homocysteine concentration in a large cohort of unselected outpatients. AB - OBJECTIVE: To evaluate the association between gamma-glutamyl transferase (GGT) and homocysteine in plasma. METHODS: Combined results of GGT, total homocysteine, vitamin B12, folate and creatinine levels were analyzed in outpatients referred for laboratory testing over the past 5 years. RESULTS: Cumulative results for homocysteine, GGT, B12, creatinine, folate could be retrieved for 449 outpatients > 35 years old over the study period. The concentration of homocysteine significantly increased among tertiles of GGT activity. In multivariable linear regression analysis, GGT activity was independently associated with homocysteine concentration, independent of age, gender, folate, vitamin B12 and serum creatinine levels. DISCUSSION: Plasma GGT activity might be a useful means to predict homocysteine concentration in the general population. PMID- 18421186 TI - Relationship between metabolic syndrome and sleep-disordered breathing in patients with cardiovascular disease--metabolic syndrome as a strong factor of nocturnal desaturation. AB - OBJECTIVE: Metabolic syndrome (MetS) is one of the coronary risk factors for cardiovascular disease and is closely related with sleep-disordered breathing (SDB). Our aim in this study was to estimate the relationship between sleep related breathing events and coronary risk factors, including MetS. METHODS: We determined the prevalence of MetS in 195 patients with cardiovascular disease. Based on Japanese MetS criteria, 56 patients had MetS (Group A), whereas 139 patients did not (Group B). We assessed SDB and sleep-related breathing events, including nocturnal desaturation, in both groups using a type 3 apparatus (Morpheus); Teijin Pharma Limited, Tokyo, Japan). RESULTS: Seventy-seven percent of the patients with MetS (43/56) met the criteria for SDB based on apnea hypopnea index (AHI). The AHI value was significantly greater in Group A than in Group B (30.1 +/- 19.0/hr vs. 17.7 +/- 14.7/hr; p < 0.001). Nocturnal oximetry showed that Group A spent a greater percentage of time at pulse-oximetric oxygen saturation below 90% (CT 90) than did Group B (10.6 +/- 13.2% vs. 5.0 +/- 12.5%; p < 0.01). On multivariate logistic regression analysis for CT 90, MetS showed that the odds ratio was 2.629 (95% confidence interval: 1.259-5.592; p = 0.011). CONCLUSION: These results suggest that SDB is common in cardiovascular patients with MetS. Patients with MetS frequently experience a sleep-related breathing event. Compared with the incidence of apnea hypopnea, MetS is an equivalently strong factor of nocturnal desaturation in patients with cardiovascular disease. PMID- 18421187 TI - Efficacy of risedronate in Japanese male patients with primary osteoporosis. AB - OBJECTIVE: Although osteoporosis in men previously was relatively neglected, bisphosphonates have been strongly suggested as potent therapeutic agents. However, there are few studies on the effects of risedronate in male osteoporosis, especially in Japanese with primary osteoporosis. The aim of our study was to prospectively evaluate the effects of risedronate on bone mineral density (BMD) and bone turnover in Japanese male patients. METHODS: According to the therapeutic regimen, the subjects were divided into two groups (group A, 22 with risedronate; group B, 10 without risedronate). During a one-year study duration, we measured bone-specific alkaline phosphatase (BAP) and serum N terminal telopeptide of type I collagen (NTx) every 3 months, and BMD at 7 sites by dual-energy X-ray absorptiometry every 6 months. PATIENTS: The subjects were 32 Japanese male patients with untreated primary osteoporosis. RESULTS: In group A, but not in group B, BMD was significantly increased at the lumbar spine both at 6 months and 12 months, and at the femoral neck at 12 months, compared with baseline. Likewise, in group A, but not in group B, both BAP and NTx were significantly decreased at all time points measured (3 months, 6 months, and 12 months), compared with baseline. CONCLUSION: These results confirmed the beneficial effects of risedronate upon increasing BMD and reducing bone turnover markers in Japanese male patients with primary osteoporosis, comparable to those previously reported in postmenopausal patients with osteoporosis. PMID- 18421188 TI - Clinical characteristics of 852 patients with subacute thyroiditis before treatment. AB - OBJECTIVE: Subacute thyroiditis (SAT) is a transient inflammatory disease of the thyroid. We evaluated the clinical characteristics based on laboratory and imaging studies in patients with SAT before treatment. PATIENTS: We reviewed the medical records of 852 patients (107 men and 745 women) with SAT who visited our thyroid clinic at Kuma Hospital from 1996 through 2004. RESULTS: SAT developed most often in female patients aged 40 to 50 years, with significant seasonal clusters during summer to early autumn. While the rates of any virus infections and diseases did not differ from those in the general population, recurrent episodes of SAT at intervals of 13.6+/-5.6 years accounted for 1.6% of all cases. At the onset of SAT, 28.2% of patients had temperatures greater than 38 degrees C and typical symptoms associated with thyrotoxicosis developed in more than 60% of patients. Before treatment, most of the abnormal laboratory findings associated with thyrotoxicosis, inflammation, and liver dysfunction reached peak levels within 1 week after onset. Ultrasound examination showed that half of the patients with unilateral thyroid pain presented with bilateral hypoechogenic area in the thyroid and the rate of bilateral hypoechogenic area tended to increase 2 months after onset. CONCLUSION: Laboratory studies of thyroid dysfunction and inflammation related to SAT presented peak levels within 1 week after onset. PMID- 18421189 TI - Long-term treatment of steroid-dependent myasthenia gravis patients with low-dose tacrolimus. AB - OBJECTIVE: To examine the long-term effects of tacrolimus in steroid-dependent myasthenia gravis (MG) patients. PATIENTS AND METHODS: We administered tacrolimus at 3 mg/day to 10 generalized MG patients presented with clinical worsening by a reduction in dose of prednisolone. The effects of tacrolimus were assessed by using the MG activities of daily living (MG-ADL) profile and the post intervention status criteria provided by the Myasthenia Gravis Foundation of America (PSC-MGFA). RESULTS: Seven patients were able to use tacrolimus without serious adverse effects for 1.0-5.1 years (mean 3.1 years). Further, its administration improved myasthenic symptoms to the level of pharmacologic remission or minimal manifestations of PSC-MGFA in 5 patients and made it possible to discontinue prednisolone administration in 4 of those 5. However, despite improvements caused by tacrolimus, the reduction in dose of prednisolone caused worsening of symptoms in another 2 patients. In addition, blood trough levels of tacrolimus lower than the recommended range were effective to maintain long-term improvements in 2 patients. CONCLUSIONS: Administration of tacrolimus induced long-term improvements and enabled replacement of prednisolone in patients with intractable steroid-dependent MG. PMID- 18421190 TI - Reversible bone marrow dysplasia in patients with systemic lupus erythematosus. AB - OBJECTIVE: Several reports of bone marrow dysplasia in patients with systemic lupus erythematosus (SLE) have been published. However, the reports are restricted primarily to descriptions of the erythroid lineage; no follow-up studies have been reported, and the clinical significance of the dysplasias is unknown. Therefore, in the present study, the dysplasias noted in bone marrow aspirates obtained from SLE patients were characterized. PATIENTS AND METHODS: The smears of bone marrow aspirates obtained from 17 SLE patients who had bone marrow aspiration due to cytopenia (WBC < 1,500/microl, or Hb < 10.5 g/dl, or platelet count < 10 x 10(4)/microl) were examined retrospectively. Of the 17 patients, 4 had a repeat bone marrow aspiration during follow-up. Clinical and laboratory data were obtained from the medical records. RESULTS: Of the 17 SLE patients, 12 had dysplasias, including: erythroid cell multinuclearity (trinuclear or more) (5 patients), megaloblastoid changes (4), pseudo-Pelger abnormalities (6), annular nuclear myeloid cells (2), separated nuclear megakaryocytes (4), and micromegakaryocytes (5). In the 4 patients who had follow up bone marrow aspiration, these dysplasias were correlated with disease activity; some abnormalities disappeared with remission of SLE. Diffuse proliferative glomerulonephritis (3 patients) and cerebral lupus/neuropsychiatric lupus (4 patients) were seen only in patients with dysplasia. CONCLUSION: This study found that bone marrow dysplasia can be observed in all lineage cells of SLE patients, and that the dysplasia is reversible during the course of the disease. The presence of dysplasias appears to be associated with disease severity. PMID- 18421191 TI - Intrahepatic splenosis with severe iron deposition presenting with atypical magnetic resonance images. AB - An intrahepatic mass was incidentally found in a 41-year-old man with a history of a traffic accident injury which resulted in removal of a ruptured spleen. Hepatic splenosis was considered in the differential diagnosis but magnetic resonance imaging showed hypointensity on T2-weighted images, atypical for normal spleen. Histologically, the mass showed sinusoidal structures and lymphoid follicular aggregates. Immunohistochemical study showed that the phenotype of the vascular lining cells was CD8-positive, CD31-positive, and CD34 negative, the pattern diagnostic for ectopic spleen. In addition, severe iron deposition was histologically demonstrated, which was considered as the cause of the hypointense T2-weighted images. PMID- 18421192 TI - Type 1 diabetes mellitus provoked by peginterferon alpha-2b plus ribavirin treatment for chronic hepatitis C. AB - A 51-year-old man developed type 1 diabetes mellitus following 24 weeks of treatment with recombinant alpha-2b peginterferon plus ribavirin for chronic hepatitis C. Pancreatic autoantibody tests were negative before the start of therapy, but a significant increase in glutamic acid decarboxylase (GAD) antibody titer was seen after 24 weeks of treatment. Six months after the onset of type 1 diabetes mellitus, the patient continues to receive 40 units of insulin daily. The clinical course suggested that recombinant alpha-2b peginterferon plus ribavirin provoked type 1 diabetes mellitus, therefore, in patients who are candidates for interferon therapy the presence of pancreatic autoantibodies and the fasting plasma glucose level should be investigated before and during treatment. PMID- 18421193 TI - Symptomatic hypercalcemia due to painless thyroiditis after unilateral adrenalectomy in a patient with Cushing's syndrome. AB - We report a 53-year-old woman with Cushing's syndrome due to an adrenocortical adenoma, who underwent unilateral adrenalectomy and developed symptomatic hypercalcemia during the thyrotoxic period of painless thyroiditis, while tapering off a daily supplemented dose of cortisol. A study of patients with thyrotoxicosis and hypoadrenalism at our institute revealed that mild hypercalcemia was present in 9.9% of those with thyrotoxicosis and 5.0% of those with hypoadrenalism. The present case suggests that the simultaneous occurrence of thyrotoxicosis and hypoadrenalism may lead to overt hypercalcemia due to a synergistic increase in bone resorption and impaired urinary excretion of calcium. PMID- 18421194 TI - Differential diagnosis and appropriate treatment of four thyrotoxic patients with Graves' disease required to take amiodarone due to life-threatening arrhythmia. AB - We report the treatment of four thyrotoxic patients. Two were cases of type I amiodarone-induced thyrotoxicosis (AIT) treated with methimazole. The third Graves' disease patient, who became hypothyroid 25 years after subtotal thyroidectomy, developed type II AIT. Furthermore, one case with heart failure and ventricular tachycardia, who developed an adverse reaction to antithyroid agents and was prescribed amiodarone, underwent total thyroidectomy. The clinical course was uneventful, and the patient is doing well. Since amiodarone contains a large amount of iodine, it is frequently difficult to make a differential diagnosis. Surgical treatment of Graves' disease patients is recommended when immediate control of hyperthyroidism and heart failure is required. PMID- 18421195 TI - Lymphoid interstitial pneumonia associated with common variable immunoglobulin deficiency. AB - The patient was a 30-year-old woman with a disease of common variable immunodeficiency. She was admitted to our hospital because of abnormal findings on her chest radiographs in an annual health screening. Chest computed tomography showed multiple reticulonodular infiltrates in both lower lung fields. The surgical lung biopsy specimen demonstrated involvement of mature small lymphocytes with a mixture of other mononuclear cells in the lung parenchyma and bronchiolar walls. The patient was diagnosed to have lymphoid interstitial pneumonia. The patient was treated with prednisolone and intravenous supplement of immunoglobulin, resulting in radiographically and clinically stable disease. PMID- 18421196 TI - Insidious pulmonary fibrosis occurring at the hypoperfusion area in a patient with chronic pulmonary thromboembolism. AB - A 69-year-old woman who had been diagnosed with chronic pulmonary thromboembolism (CPTE) developed ground glass opacities in the right lung where perfusion scintigraphy showed defects of the bloodstream. Bronchoalveolar lavage fluid (BALF) showed that some macrophages had phagocytosed hemosiderin. Video-assisted thoracoscopic surgical biopsy revealed lung fibrosis, narrowing of the pulmonary artery and organization of many cholesterol granulomas. We hypothesized that hyperperfusion of the bronchial artery, which occurred to compensate for hypoperfusion of the pulmonary artery, induced alveolar hemorrhage following thrombolytic and anticoagulation treatments, and that the degraded products from blood cells induced the formation of lung fibrosis and cholesterol granulomas. PMID- 18421197 TI - Severe Legionnaires' disease successfully treated using a combination of fluoroquinolone, erythromycin, corticosteroid, and sivelestat. AB - The patient was a 67-year-old man with diabetes mellitus who had been to a hot spring spa a few days before his admission. The diagnosis of Legionella pneumonia was made using a urinary antigen assay. Intravenous pazufloxacin and oral clarithromycin were started. However, despite these treatments, he developed acute respiratory distress syndrome (ARDS). He was administered the combination of intravenous pazufloxacin and erythromycin, corticosteroid, and sivelestat for two weeks. Then he was successfully recovered. The outcome suggests that treatment with corticosteroid and sivelestat, in addition to a combination of appropriate anti-Legionella antibiotics, should be considered for patients with severe Legionella pneumonia with ARDS. PMID- 18421198 TI - Desquamative interstitial pneumonia with a remarkable increase in the number of BAL eosinophils. AB - A 57-year old man with desquamative interstitial pneumonia (DIP) showed a marked increase in eosinophils in the bronchoalveolar lavage (BAL) fluid. The patient was referred to our hospital for abnormal shadows on his chest X-ray with no symptoms in May 2007. Computed tomography (CT) showed patchy, peripheral predominate ground-glass opacity. The BAL fluid revealed an increase of the total number of cells, including markedly elevated levels of eosinophils (62.1%), in contrast with only a slight increase of peripheral blood eosinophils, or minimal eosinophils in the alveolar spaces and interstitium of the thoracoscopic lung biopsy specimen. Since the specimens showed findings compatible with a DIP pattern, we diagnosed the patient with DIP. Although it is a rare entity, we should therefore consider DIP in the differential diagnosis when we encounter patients with a marked increase in the number of BAL eosinophils. PMID- 18421199 TI - Rapidly progressive interstitial pneumonia associated with clinically amyopathic dermatomyositis successfully treated with polymyxin B-immobilized fiber column hemoperfusion. AB - Amyopathic dermatomyositis (ADM) is a clinical subtype of dermatomyositis, characterized by the absence of motor weakness and the presence of normal muscle enzyme levels. ADM is sometimes accompanied by interstitial pneumonia that shows a rapid progressive course associated with a poor prognosis. We describe a 70 year-old man who presented rapidly progressive interstitial pneumonia associated with clinically ADM (C-ADM); he was successfully treated with polymyxin B immobilized fiber column (PMX) hemoperfusion. PMID- 18421200 TI - Successful treatment of chronic myeloproliferative disease-unclassifiable (CMPD U) with no chromosomal abnormalities by imatinib mesylate. AB - We report a chronic myeloproliferative disease-unclassifiable (CMPD-U) patient who achieved hematological remission following imatinib mesylate (imatinib). Chromosomal and molecular analyses demonstrated no genetic abnormalities of c abl, bcr-abl, c-kit or platelet-derived growth factor receptor (PDGFR) genes from hematopoietic cells. Although there has been one report of CMPD-U patient with chromosomal abnormalities of the PDGFR gene having complete hematologic responses upon treatment with imatinib, there have not been similar reports of patients without chromosomal abnormalities. This is the first case report of a CMPD-U patient with no chromosomal abnormalities who completely responded to treatment with imatinib. PMID- 18421201 TI - Unilateral hearing disturbance could be an isolated manifestation prior to ipsilateral anterior inferior cerebellar artery infarction. AB - A 50-year-old man presented with a sudden onset of right-sided hearing disturbance. His hearing disturbance improved gradually, however, dysarthria, right-sided facial weakness and dysesthesia, and gait disturbance was developed 11 days after the onset of hearing disturbance. MR imaging revealed fresh infarctions of the right dorsolateral pons and middle cerebellar peduncle localized in the territory of anterior inferior cerebellar artery. Unilateral hearing disturbance could be an isolated manifestation prior to ipsilateral anterior inferior cerebellar artery infarction. PMID- 18421202 TI - Intradural extramedullary tuberculoma of the thoracic spine: paradoxical response to antituberculous therapy. AB - Intradural extramedullary tuberculoma of the spinal cord (IETSC) is a rare complication of tuberculosis, which can occur as a paradoxical response to antituberculous therapy. A 46-year-old woman with tuberculosis meningitis developed an acute sensory disturbance and paraplegia eight weeks after the antituberculous treatment was started. MRI revealed a cystic lesion at the Th 2 and 3 vertebrae levels, and continuous dural thickening. Laminectomy was performed; soft granulomas were unexpectedly observed inside the dura matter. After the operation, the patient experienced progressive improvement in motor strength. IETSC should be known as rare but possible complication of tuberculous meningitis. PMID- 18421203 TI - Nosocomial acinetobacter genomic species 13 TU endocarditis following an endoscopic procedure. AB - We report a rare case of prosthetic valve endocarditis caused by Acinetobacter genomic species 13 TU. This patient had rheumatic heart disease and received prosthetic mitral valve replacement eleven years previously. He was admitted due to tarry stool. Endoscopic procedure showed two gastric ulcers and some mucous breaks at the distal esophagus. He had a fever on the eleventh hospital day. Persistent Acinetobacter bacteremia was noted with conjunctiva hemorrhage. The pathogen was identified as Acinetobacter genomic species 13 TU by PCR-based method. According to his whole course of disease, the most possible portal of entry was via the endoscopic procedure. PMID- 18421204 TI - Hepatic cystic echinococcosis with specific CT findings. AB - A hepatic central bisegmentectomy was performed on a 36-year-old Iranian man with suspected cystic echinococcosis. Hepatic computed tomography (CT) scan findings showed a large cystic lesion, which included many small round shaped cystic lesions. The diagnosis of hepatic cystic echinococcosis was confirmed during surgery. The aforementioned CT scan findings may be specific findings for cystic echinococcosis, in spite of a low appearance rate. PMID- 18421205 TI - Usher's syndrome. PMID- 18421206 TI - Venous thrombosis associated with a large uterine myoma. PMID- 18421207 TI - Giant coronary aneurysm fistulous connection to the right ventricle. PMID- 18421208 TI - CARTO images after heart transplantation. PMID- 18421209 TI - Hypopituitarism caused by bilateral internal carotid artery aneurysms with a carotid-cavernous fistula. PMID- 18421210 TI - Magnetic resonance images in rhombencephalitis due to Listeria monocytogenes mimicking Wernicke encephalopathy. PMID- 18421211 TI - Gene expression profiling distinguishes between spontaneous and radiation-induced rat mammary carcinomas. AB - The ability to distinguish between spontaneous and radiation-induced cancers in humans is expected to improve the resolution of estimated risk from low dose radiation. Mammary carcinomas were obtained from Sprague-Dawley rats that were either untreated (n = 45) or acutely gamma-irradiated (1 Gy; n = 20) at seven weeks of age. Gene expression profiles of three spontaneous and four radiation induced carcinomas, as well as those of normal mammary glands, were analyzed by microarrays. Differential expression of identified genes of interest was then verified by quantitative polymerase chain reaction (qPCR). Cluster analysis of global gene expression suggested that spontaneous carcinomas were distinguished from a heterogeneous population of radiation-induced carcinomas, though most gene expressions were common. We identified 50 genes that had different expression levels between spontaneous and radiogenic carcinomas. We then selected 18 genes for confirmation of the microarray data by qPCR analysis and obtained the following results: high expression of Plg, Pgr and Wnt4 was characteristic to all spontaneous carcinomas; Tnfsf11, Fgf10, Agtr1a, S100A9 and Pou3f3 showed high expression in a subset of radiation-induced carcinomas; and increased Gp2, Areg and Igf2 expression, as well as decreased expression of Ca3 and non-coding RNA Mg1, were common to all carcinomas. Thus, gene expression analysis distinguished between spontaneous and radiogenic carcinomas, suggesting possible differences in their carcinogenic mechanism. PMID- 18421212 TI - The utility of two daytime icodextrin exchanges to reduce dextrose exposure in automated peritoneal dialysis patients: a pilot study of nine patients. AB - BACKGROUND: The use of dextrose-containing solutions in peritoneal dialysis (PD) is thought to be associated with glucose-related toxicity both to the peritoneal membrane and systemically. There has, therefore, been considerable interest in minimizing the use of dextrose exposure during PD. The present study was designed to explore the use of icodextrin in patients with high/high-average transporter characteristics for two exchanges per day to minimize glucose exposure. METHODS: We performed a 6-month prospective cohort study using two icodextrin exchanges per day in a group of high/high-average transporters maintained on automated PD. Icodextrin levels, serum sodium levels, and glucose exposure were measured at baseline, 3 and 6 months. RESULTS: Nine patients completed the study protocol. While the total volume of PD solution remained the same, there was a reduction in mean glucose exposure from a baseline mean value of 410 +/- 75 to 275 +/- 57 g/day at 3 months and 300 +/- 75 g/day at 6 months. Serum icodextrin levels rose from a baseline mean of 345 +/- 145 to 615 +/- 120 mg/dl at 3 months and 620 +/- 108 mg/dl at 6 months. Serum sodium levels remained stable. CONCLUSION: The use of two (double) icodextrin exchanges in high/high-average transporters on PD can contribute to reduction in glucose exposure for patients maintained on automated PD and appears to be safe. PMID- 18421213 TI - Bone mineral density in end-stage renal disease patients: association with wasting, cardiovascular disease and mortality. AB - BACKGROUND: Bone and mineral disorders may contribute to extraosseous ossifications and cardiovascular disease (CVD) in end-stage renal disease (ESRD) patients. We have investigated the relationship between bone mineral density (BMD) and inflammation, wasting, CVD and mortality in ESRD patients. METHODS: BMD (dual energy X-ray absorptiometry) and biochemical, nutritional and inflammatory markers were assessed in 277 incident ESRD patients (GFR 7.1 +/- 0.2 ml/min) who were then followed prospectively for a mean of 27 (range 1-60) months. Carotid plaques were determined in 103 patients. RESULTS: Patients with carotid plaques, clinical manifestation of CVD and wasting (assessed by subjective global assessment) had significantly lower BMD than their counterparts. Low BMD was associated with high all-cause and cardiovascular mortality. Even after adjustment for several confounders and risk factors, all-cause (HR = 2.1, CI: 1.1 3.9, p = 0.02) and cardiovascular (HR = 2.8, CI: 1.2-6.3, p = 0.02) mortality remained significantly associated with low BMD. CONCLUSIONS: Low BMD is associated with wasting and CVD, and is an independent predictor of all-cause and cardiovascular mortality in ESRD patients. PMID- 18421214 TI - Cytokine dysregulation in chronic kidney disease: how can we treat it? AB - As the kidney is the major site for elimination of many cytokines, the delicate equilibrium of pro-inflammatory cytokines and their inhibitors is clearly dysregulated in chronic kidney disease (CKD) patients. The consequences of the altered immune response in uremia lead to a state of persistent inflammation which is highly prevalent among CKD patients and is linked to complications such as the development of protein-energy wasting and atherosclerotic vascular disease. The present review aims at reviewing this complex orchestration of uremic cytokines beyond the well-studied interleukin-6 and tumor necrosis factor alpha. Finally, we update our current understanding on anti-inflammatory treatment strategies in CKD patients, including nutritional and lifestyle measurements, pharmacological intervention and specific anticytokine strategies targeting the dialytic procedure. PMID- 18421216 TI - Leave 'em alone - why continuous variables should be analyzed as such. PMID- 18421217 TI - Fasting insulin levels and cognitive decline in older women without diabetes. AB - BACKGROUND: Type 2 diabetes has been associated with an increased risk of dementia. To assess possible independent effects of insulin, we investigated the relation of insulin levels to cognitive decline in nondiabetic women. METHODS: Fasting plasma insulin levels were measured in mid-life in 1,416 nondiabetic Nurses' Health Study participants, who also completed cognitive testing that began 10 years later (current age: 70-75 years). Over 4 years, 3 assessments of general cognition, verbal memory, category fluency and attention were administered. Primary outcomes were the Telephone Interview for Cognitive Status (TICS) performance, the global score (average of all tests) and verbal memory (average of verbal recall tests). Linear mixed-effects models were used to calculate the association between insulin and cognitive decline. RESULTS: Higher insulin levels were associated with a faster decline on the TICS and verbal memory. For analysis, batch-specific quartiles of insulin levels were constructed. Compared to the lowest quartile, adjusted differences in the annual rates of decline (with 95% CI values in parentheses) for the second, third and fourth quartiles were: TICS, -0.06 (-0.16, 0.03), -0.14 (-0.24, -0.04), and -0.09 (-0.19, 0.01) points (p trend = 0.04); verbal memory, -0.01 (-0.04, 0.02), -0.05 (-0.08, -0.02), and -0.02 (-0.05, 0.01) units (p trend = 0.02). These associations remained after multivariable adjustment. CONCLUSIONS: Our study provides evidence for a potential role of higher fasting insulin levels in cognitive decline, possibly independent of diabetes. PMID- 18421218 TI - The National Registry of Veterans with amyotrophic lateral sclerosis. AB - BACKGROUND: The Department of Veterans Affairs (VA) Cooperative Studies Program has established a National Registry of Veterans with Amyotrophic Lateral Sclerosis (ALS). This article describes the objectives, methods, and sample involved in the registry. METHODS: United States military veterans with ALS were identified through national VA electronic medical record databases and nationwide publicity efforts for an enrollment period of 4 1/2 years. Diagnoses were confirmed by medical record reviews. Registrants were asked to participate in a DNA bank. Follow-up telephone interviews are conducted every 6 months to track participants' health status. RESULTS: As of September 30, 2007, 2,400 veterans had consented to participate in the registry, 2,068 were included after medical record review, 995 were still living and actively participating, and 1,573 consented to participate in the DNA bank. 979 participants had been enrolled in the registry for at least 1 year, 497 for at least 2 years, and 205 for at least 3 years. Fourteen studies have been approved to use registry data for epidemiological, observational, and interventional protocols. CONCLUSION: This registry has proven to be a successful model for identifying large numbers of patients with a relatively rare disease and enrolling them into multiple studies, including genetic protocols. PMID- 18421220 TI - New insights into the biology of peritoneal mesothelial cells: the roles of epithelial-to-mesenchymal transition and cellular senescence. AB - Ultrafiltration failure due to dysfunction of the peritoneum as a dialyzing organ is a major clinical limitation of peritoneal dialysis. It is increasingly clear that mesothelial cells play an important role in fibrogenesis and vasculopathy that underlie peritoneal membrane dysfunction. New and extensively studied aspects of peritoneal mesothelial cell biology include epithelial-to-mesenchymal transition and cellular senescence. We discuss the potential significance of these processes for the peritoneal membrane function. PMID- 18421219 TI - Genes and Environmental Exposures in Veterans with Amyotrophic Lateral Sclerosis: the GENEVA study. Rationale, study design and demographic characteristics. AB - Recent reports of a potentially increased risk of amyotrophic lateral sclerosis (ALS) for veterans deployed to the 1990-1991 Persian Gulf War prompted the Department of Veterans Affairs to establish a National Registry of Veterans with ALS, charged with the goal of enrolling all US veterans with a neurologist confirmed diagnosis of ALS. The Genes and Environmental Exposures in Veterans with ALS study (GENEVA) is a case-control study presently enrolling cases from the Department of Veterans Affairs registry and a representative sample of veteran controls to evaluate the joint contributions of genetic susceptibility and environmental exposures to the risk of sporadic ALS. The GENEVA study design, recruitment strategies, methods of collecting DNA samples and environmental risk factor information are described here, along with a summary of demographic characteristics of the participants (537 cases, 292 controls) enrolled to date. PMID- 18421221 TI - Serum proteomic profiling associated with immune system impaired by stress using ProteinChip technology. AB - We set out to use proteomic profiling as a means of defining serum peptides that are indicative of a stress response related to an impact on the immune system. Two types of mouse models with the immune response impaired under stress (restraint stress and thermal stress) comprised the stressed groups (n = 10 in each group), while mice without stress represented the control group (n = 10). ProteinChip technology (surface-enhanced laser desorption/ionization time-of flight mass spectrometry) was used to investigate the serum proteomic profiles. Histopathological examination of spleens from the stressed and control groups was performed to select appropriate individuals. Compared with the spleens of normal mice, those of mice subjected to restraint and thermal stress had decreased white pulp and lymphoid nodules, and their distribution and structure changed irregularly. Pathological changes occurred in all individuals in both stressed groups. The m/z values of the protein peaks ranged from 1,500 to 50,000 daltons and were mainly in the 2,000- to 20,000-dalton range. One hundred and fifty protein peaks were detected in the three groups. Four downregulated universal stress protein peaks with m/z of 4,389, 5,341, 5,526, and 6,252 were finally selected. Unexpectedly, no upregulated universal stress protein peaks were detected. These results suggested that impairment of the immune system results from inhibition of growth-promoting factors associated with the immune system. Identified protein peaks may be biomarkers of the impaired immune system under stress. PMID- 18421222 TI - Editorial. PMID- 18421223 TI - Lipid-bloated subretinal microglial cells are at the origin of drusen appearance in CX3CR1-deficient mice. AB - Drusen, the white yellowish deposits that can be seen in funduscopy, are a hallmark of age-related macular degeneration. Histologically, drusen are believed to be dome-shaped or more confluent lipid accumulations between the retinal pigment epithelium and the choriocapillaries. Recent advances in mouse funduscopy have revealed the presence of drusen-like structures in chemokine knockout animals in the absence of sizeable dome-shaped material below the retinal pigment epithelium. We show that aged CX3CR1-/- mice present with drusen-like appearance in funduscopy that is associated with a progressive age-related microglial cell accumulation in the subretinal space. We demonstrate that the anatomical equivalent of the drusen-like appearance in these mice are lipid-bloated subretinal microglial cells rather than subretinal pigment epithelium deposits [Combadiere C, et al: J Clin Invest 2007;117:2920-2928]. PMID- 18421224 TI - Lack of alphavbeta5 integrin receptor or its ligand MFG-E8: distinct effects on retinal function. AB - BACKGROUND/AIMS: Diurnal phagocytosis of spent photoreceptor outer segment fragments by the retinal pigment epithelium (RPE) is critical for vision. We recently identified an important role for alphavbeta5 integrin receptors and their ligand Milk fat globule-EGF factor 8 (MFG-E8) in RPE phagocytosis. METHODS: We compared RPE phagocytosis and retinal function between mice deficient in alphavbeta5 integrin receptors and mice deficient in the secreted integrin ligand MFG-E8. RESULTS: Both beta5-/- and MFG-E8-/- mice exhibit the same phagocytic defect: RPE cells retain basal uptake activity but completely lack the burst of phagocytic activity as well as the rhythmic activation of Mer tyrosine kinase that follow circadian photoreceptor shedding in wild-type RPE. Strikingly, electroretinogram photoresponses decline with age only in beta5 -/- but not in MFG-E8-/- retina. CONCLUSION: These results identify a critical role of alphavbeta5 integrin receptors and their ligand MFG-E8 in synchronizing retinal phagocytosis. Additionally, we show that lack of alphavbeta5 receptors and MFG-E8 ligand have distinct consequences for retinal function. These intriguing results suggest that loss of phagocytic rhythm is not solely responsible for the age related blindness of beta5-/- mice. PMID- 18421225 TI - Ccl2/Cx3cr1-deficient mice: an animal model for age-related macular degeneration. AB - BACKGROUND/AIMS: Senescent Ccl2-/- mice develop cardinal features of human age related macular degeneration (AMD). Loss-of-function single-nucleotide polymorphisms within CX3CR1 are associated with AMD. METHODS: We generated Ccl2-/ /Cx3cr1-/- [double-knockout (DKO)] mice and evaluated the eyes using fundoscopy routine histology, immunochemistry, biochemistry and proteomics. RESULTS: At 6 weeks old, all DKO mice developed AMD-like retinal lesions such as abnormal retinal pigment epithelium cells, drusen, photoreceptor atrophy and choroidal neovascularization, which progressed with age and reversed with high omega-3 long chain polyunsaturated fatty acid diet. N-retinylidene-N-retinylethanolamine (A2E), a major lipofuscin fluorophore, illustrated by an emission peak at approximately 600 nm, was significantly higher in DKO retinal pigment epithelium. Decreased ERp29 was found in the retina of DKO mice. CONCLUSION: A broad spectrum of AMD pathologies with early onset and high penetrance in these mice implicate certain chemokines, A2E and endoplasmic reticulum proteins in AMD pathogenesis. PMID- 18421226 TI - Animal models of Vogt-Koyanagi-Harada disease (sympathetic ophthalmia). AB - BACKGROUND/AIMS: We aimed to establish the animal model of Vogt-Koyanagi-Harada disease (VKH). METHODS: Rats, Akita dogs and monkeys were immunized with the peptides derived from tyrosinase-related protein 1. Each experimental animal was examined clinically and histologically. RESULTS AND CONCLUSION: The rats developed ocular and extraocular inflammation homologous to human VKH by immunization of tyrosinase-related protein 1. The Akita dogs also developed the autoimmune disease almost similar to the spontaneously emerging dog VKH equivalent to human VKH. The monkeys developed similar clinical findings as human VKH. These models may serve as human VKH models. PMID- 18421227 TI - Experimental melanin-induced uveitis: experimental model of human acute anterior uveitis. AB - Experimental melanin-protein-induced uveitis (EMIU), which is also known as experimental autoimmune anterior uveitis, was first described in 1993 by Broekhuyse et al. This experimental uveitis may be induced in certain inbred and outbred rat strains by immunization with bovine ocular melanin. The inflammation shares clinical features with human acute anterior uveitis. The duration of the first episode is approximately 1 month. Spontaneous recovery to a near normal clinical state is the rule, but multiple recurrences are common. Slit-lamp biomicroscopic examination reveals a florid anterior-chamber reaction, with formation of a retro-iridal empyema, fibrin clots and posterior synechiae. At a microscopic level, leukocytic infiltration is first observed in the anterior uvea. Although this tissue remains the site of maximum inflammation throughout an attack, in severe cases, limbitis, vitritis and choroiditis are also observed. Abrogation of EMIU occurs after treatment with anti-CD4 antibody, indicating that the uveitis is controlled by CD4-positive T cells. Several research groups have used EMIU to investigate various aspects of the pathogenesis of acute anterior uveal inflammation, including the participation of different leukocyte subsets, the expression of cell adhesion molecules, cytokines, chemokines and nitric oxide, the role of complement and the impact of apoptosis. In addition, EMIU has also been used to evaluate various biologic interventions with potential implications for the treatment of human disease. PMID- 18421228 TI - Rat models of autoimmune uveitis. AB - Experimental autoimmune uveitis (EAU) in Lewis rats is a well-established model for human uveitis. During the last years we used this model to demonstrate extraocular induction of uveitis by antigenic mimicry of environmental antigens with retinal autoantigen and investigated the migration and intraocular reactivation of autoreactive green fluorescent protein (GFP)+ T cells. We could also elaborate several differences between EAU induced with S-antigen peptide PDSAg or R14, a peptide derived from interphotoreceptor retinoid-binding protein, suggesting two differently regulated diseases in the same rat strain. R14 mediated EAU in Lewis rats has been shown to relapse, thus we have a new model to test therapeutic approaches in an ongoing immune response instead of just preventing disease. Finally, we show antigenic mimicry of PDSAg and an HLA-B peptide for oral tolerance induction. After the successful first therapeutic trial this approach will now proceed with international multicenter clinical trials. PMID- 18421229 TI - Drainage of fluorescent liposomes from the vitreous to cervical lymph nodes via conjunctival lymphatics. AB - The use of liposomes as carriers for the delivery of biologically active molecules into the eye is of major interest. Indeed, encapsulation of biologically active molecules in liposomes may increase their bioavailability and may induce a sustained release, thus avoiding repeated intraocular injections and reducing side effects. We describe here the fate of rhodamine-conjugated liposomes (Rh-Lip) injected into the vitreous of normal Lewis rats. Twenty-four hours after intravitreal injection fluorescent liposomes were detected in the vitreous, the inner layer of the retina and to a lesser extent in the anterior segment of the eye. In addition, numerous Rh-Lip were also observed in the episclera and conjunctival stroma, in conjunctival lymphatic vessels and cervical lymph nodes (LN) draining the conjunctiva and the eye. In the LN, Rh-Lip were taken up by resident macrophages adjacent to CD4+ and CD8+ T cells. Thus, intravitreal injection of anti-inflammatory drugs loaded in liposomes could modulate the ocular immune microenvironment. In addition the passage of drugs into the cervical LN could alter the immune status of these LN and contribute to the regulation of intraocular inflammation. Our results suggest that this phenomenon should be taken into account to design new therapies based on intraocular drug administration. PMID- 18421230 TI - Equine recurrent uveitis--a spontaneous horse model of uveitis. AB - Equine recurrent uveitis (ERU) is an autoimmune disease that occurs with a high prevalence (10%) in horses. ERU represents the only reliable spontaneous model for human autoimmune uveitis. We already identified and characterized novel autoantigens (malate dehydrogenase, recoverin, CRALBP) by analyzing the autoantibody-binding pattern of horses affected by spontaneous recurrent uveitis (ERU) to the retinal proteome. CRALBP also seems to be relevant to human autoimmune uveitis. Proteomic screening of vitreous and retinal samples from ERU diseased cases in comparison to healthy controls has led to the identification of a series of differentially regulated proteins, which are functionally linked to the immune system and the maintenance of the blood-retinal barrier. PMID- 18421231 TI - Evidence for extrathymic generation of regulatory T cells specific for a retinal antigen. AB - BACKGROUND: Thymic expression of a photoreceptor cell antigen, interphotoreceptor retinoid-binding protein, is known to generate regulatory T cells (T(reg)) that prevent spontaneous autoimmune disease of the retina. However, the contribution of other endogenous, tissue-specific antigens (Ags) expressed in the retina to the generation of T(reg) is uncertain. METHODS: Transgenic mice that express beta galactosidase (beta-gal) in photoreceptor cells, together with beta-gal-specific T cell receptor transgenic mice, were used to study the induction of T(reg) in vivo. RESULTS: Transgenic expression of beta-gal on the arrestin promoter led to a spontaneous immunoregulatory response that inhibited the development of immune responses to beta-gal. The regulation was transferred by CD3+4+25+ T(reg). Several strategies were then used to show that beta-gal expressed in the retina supported spontaneous, thymus-independent T(reg) development. The endogenous T(reg) also differed from the T(reg) induced by Ag inoculation into the anterior chamber of the eye. CONCLUSION: These results demonstrate that retinal expression of very small amounts of a tissue-specific Ag can generate T(reg) in the periphery. PMID- 18421232 TI - Photoreceptor mitochondrial oxidative stress in experimental autoimmune uveitis. AB - In experimental autoimmune uveitis (EAU), the macrophages infiltrate the retina during the late phase, 10-14 days after immunization with uveitogenic antigen, causing photoreceptor damage. However, prior to inflammatory cell infiltration, during the early phase (5-7 days after immunization), increased generation of reactive oxygen and nitric oxide species was observed in the photoreceptor mitochondria indicating oxidative stress. The oxidative-stress-induced nitration of photoreceptor mitochondrial proteins and peroxidation of membrane lipids led to activation and migration of microglia toward the photoreceptors. These observations suggest oxidative stress could be an initial pathologic event leading to amplification of inflammation inducing photoreceptor damage, thereby causing clinical and histologic expression of uveitis in the form of inflammatory cell infiltration. PMID- 18421233 TI - Uveitis secondary to bacterial products. AB - Bacteria are suspected contributors to several forms of immune-mediated, noninfectious forms of uveitis including that associated with ankylosing spondylitis, sarcoidosis, Behcet's disease and inflammatory bowel disease. Endotoxin (lipopolysaccharide)-induced uveitis has been a widely used model for more than 2 decades. Both rats and mice develop a transient, bilateral anterior uveitis after a systemic injection of endotoxin. Inflammation posterior to the lens is generally milder than anterior segment inflammation. The uveitis is severer if the lipopolysaccharides are injected intraocularly. The model has been invaluable in helping to identify mediators induced in the inflamed eye and in testing pharmacologic approaches to reduce eye inflammation. Muramyl dipeptide is another bacterial cell component that can induce uveitis in laboratory animals. Muramyl dipeptide is especially intriguing as a cause of uveitis because it activates the intracellular protein, Nod2, and mutations in the NOD2 gene are the cause of the autosomal dominant form of uveitis that is characteristic of Blau syndrome. Since a mutation in a gene that codes for a protein which senses a bacterial product consistently results in uveitis, it is critical to understand more fully the mechanisms by which bacterial products cause uveitis in laboratory animals. PMID- 18421234 TI - Mouse models of experimental autoimmune uveitis. AB - The mouse model of experimental autoimmune uveitis, induced by immunization of mice with the retinal protein IRBP, was developed in our laboratory 20 years ago and published in 1988. Since that time it has been adopted by many investigators and has given rise to many studies that helped elucidate genetic influences, dissect the basic mechanisms of pathogenesis and test novel immunotherapeutic paradigms. The current overview will summarize the salient features of the experimental autoimmune uveitis model and discuss its mechanisms. PMID- 18421235 TI - Pathological aspects of spontaneous uveitis and retinopathy in HLA-A29 transgenic mice and in animal models of retinal autoimmunity: relevance to human pathologies. AB - PURPOSE: A major increased risk of developing birdshot chorioretinopathy is reported in humans who are HLA-A29-positive. To better characterize this disease, an animal model of HLA-A29-associated disease was developed and the pathology arising spontaneously in these transgenic mice was compared to animal models of autoimmune uveoretinitis and to human pathology. MATERIALS AND METHODS: HLA-A2902 cDNA (A29c) was obtained from a patient suffering from birdshot retinochoroidopathy and used for transgene construct to generate HLA-A29 transgenic mice. Histopathological examination of the animal cohort was performed up to 15 months of age. It was compared with the ocular pathology developed in C57BL/6 mice and in Lewis rats immunized with retinal autoantigens. RESULTS: Aging HLA-A29 transgenic mice spontaneously developed an ocular disease with resemblance to experimental retinal-Ag-induced autoimmune ocular disease and to human pathologies shown in birdshot retinochoroidopathy, Vogt-Koyanagi-Harada and sympathetic ophthalmia. Pathogenic mechanisms could possibly be shared by these conditions. CONCLUSION: Humanized models of ocular inflammation developed in HLA class I and class II transgenic mice will help better understand the mechanisms responsible for ocular inflammation. Local control of autoimmunity in HLA-A29 positive individuals would be an important option for new therapeutic strategies. PMID- 18421236 TI - Primate model of uveoretinitis and vasculitis/experimental autoimmune uveoretinitis induced in cynomolgus monkeys by retinal s antigen. AB - PURPOSE: We aimed to describe the clinical and angiographic changes in an experimental model of autoimmune uveoretinitis and vasculitis in primates. METHODS: Six cynomolgus monkeys received a single subcutaneous immunization with 100 microg of human S antigen with complete Freund's adjuvant. RESULTS: All the animals had a bilateral long-term disease occurring usually in 1 eye approximately 4 weeks after immunization, the second eye being involved 1-5 weeks later. A cyclic course of the disease could be demonstrated by repeated fundus fluorescein angiograms. The initial and principal manifestation consisted in retinal vascular sheathing affecting veins and venules. The more severe forms showed areas of posterior uveoretinitis, dense vitritis and anterior uveitis. CONCLUSION: A single systemic injection of pure human retinal S antigen could induce a chronic and recurrent ocular disease similar to human retinal vasculitis. PMID- 18421237 TI - Anterior uveitis accompanies joint disease in a murine model resembling ankylosing spondylitis. AB - BACKGROUND: Uveitis is often associated with a systemic inflammatory disease such as ankylosing spondylitis. Our understanding of the eye's susceptibility to immune-mediated uveitis as in the apparent absence of infection has been limited by a relative lack of experimental models. Here we sought to assess whether ocular inflammation occurs in a previously described murine model of proteoglycan induced spondylitis, wherein mice develop progressive spondylitis, sacroiliitis and peripheral arthritis--features common to the clinical presentations of ankylosing spondylitis. METHODS: Using intravital microscopy we examined the ocular inflammatory response after the onset of arthritis in mice that overexpressed the T cell receptor (TCR) specific for a dominant arthritogenic epitope of cartilage proteoglycan [TCR-Tg (transgenic) mice] or BALB/c controls. RESULTS: Immunized TCR-Tg mice showed a significant increase in the number of rolling and adhering cells within the iris vasculature compared to adjuvant control mice. Cellular infiltration within the iris tissue, as assessed by intravital microscopy and histology, was also increased. Our initial temporal analysis has revealed that immunized TCR-Tg mice show a significant increase in intravascular inflammation by 2 weeks after immunization, but it diminishes at 4 weeks after immunization. CONCLUSIONS: Although these data are preliminary, this model has the potential to clarify the mechanisms accounting for the coexistence of eye and sacroiliac inflammation as occurs in patients with ankylosing spondylitis. PMID- 18421239 TI - Relevance of animal models to human uveitis. AB - Uveitis represents a wide spectrum of intraocular inflammatory conditions and includes various autoimmune and infectious etiologies. The relevance of animal models of uveitis to human diseases remains a key issue with major implications for the translational research and development of therapeutic strategies. Histopathological findings in patients with Vogt-Koyanagi-Harada disease, birdshot retinochoroidopathy and anterior uveitis are correlated to those observed in different animal models. Even though evidence based on histopathology is usually irrefutable, similar features may be due to different disease mechanisms. Analysis of triggering factors, determination of cellular populations and immune microenvironment should prevail over clinical phenotype evaluation. There is a controversy in correlating the clinical finding of nummular chorioretinal scars, commonly referred to as Dalen-Fuchs nodules, seen in the periphery of fundus in patients with chronic Vogt-Koyanagi-Harada disease with histologic observations made on such enucleated eyes. Although histopathology of the lesions consisted of focal chorioretinal scars with loss of RPE, there was no consensus about the histologic nature of the nummular chorioretinal scars, particularly whether they represent Dalen-Fuchs nodules. Based on the immunogenetic background, there may be different forms of one specific disease with variable phenotypic expression. This review discusses the importance of experimental models in the light of immunologic alterations and histopathological features in human uveitic entities. PMID- 18421238 TI - Microbial products trigger autoimmune ocular inflammation. AB - PURPOSE: Microbial products stimulate the immune system by interacting with Toll like receptors (TLR) on antigen-presenting cells. This study examined the hypothesis that microbial products, which function as TLR ligands, are playing a major role in triggering pathogenic autoimmunity. METHODS: An experimental system was developed in which microbial TLR ligands were tested in vivo for their capacity to stimulate naive CD4 cells specific against hen egg lysozyme (HEL) to become effector cells capable of inducing inflammation in eyes in which HEL is expressed. The ligands' mode of action was analyzed by determining their effects on the proliferation, acquisition of tissue-invading capacity, i.e. elevated CD49d and decreased CD62L expression, and production of interferon-gamma by the HEL-specific cells. RESULTS: All the 7 tested TLR ligands triggered ocular inflammation in the experimental system used here, with pertussis toxin surpassing all other ligands in its activities. A correlation was found between the capacity of the ligands to trigger pathogenic immunity and to stimulate the proliferation, modification of cell surface and interferon-gamma production by T cells. CONCLUSIONS: This study provides direct evidence to support the notion that microbial products are capable of triggering pathogenic autoimmunity. PMID- 18421240 TI - PlGF-1 and VEGFR-1 pathway regulation of the external epithelial hemato-ocular barrier. A model for retinal edema. AB - VEGF is considered as an important factor in the pathogenesis of macular edema. VEGF induces the rupture of the blood retinal barrier and may also influence the retinal pigment epithelial (RPE) outer retinal barrier. The aim of this work was to analyze the influence of the VEGF receptor pathways in the modulation of the RPE barrier breakdown in vitro and in vivo. The ARPE19 human junctions in culture are modulated by VEGF through VEGFR-1 but not through VEGFR-2. PlGF-1, that is a pure agonist of VEGFR-1, is produced in ARPE-19 cells under hypoxic conditions and mimics VEGF effects on the external retinal barrier as measured by TER and inulin flux. In vivo, the intravitreous injection of PlGF-1 induces a rupture of the external retinal barrier together with a retinal edema. This effect is reversible within 4 days. VEGF-E, that is a pure agonist of VEGFR-2, does not induce any acute effect on the RPE barrier. These results demonstrate that PlGF-1 can reproduce alterations of the RPE barrier occurring during diabetic retinopathy. PMID- 18421241 TI - Animal models of intraocular lymphomas. AB - Primary intraocular lymphoma is a high-grade non-Hodgkin lymphoma whose pathogenesis is still unclear. Few animal models exist in order to study this condition. Although intraocular lymphomas in humans are usually B cell lymphomas, most of these models are T cell lymphomas. Recently, a major step forward has been realized with the development of new models of intraocular B cell lymphoma. New therapeutic tools are being evaluated in these models of B cell lymphoma. We evaluate the contribution of the different animal models available to study intraocular lymphomas, and we discuss the new therapeutic strategies and their various targets in the tumor as well as in the environment, which are currently investigated through the development of these models. PMID- 18421242 TI - Optic nerve degeneration in experimental autoimmune encephalomyelitis. AB - The mechanisms of axonal and neuronal degeneration causing disability in optic neuritis and multiple sclerosis are poorly understood. Here we describe the role of mitochondria, oxidative stress and the effects of modulating antioxidant gene expression in the optic nerves of mice induced with experimental autoimmune encephalomyelitis, with a focus on long-term neuroprotection. Oxidative injury to the mitochondrion began prior to inflammatory cell infiltration and continued. It affected subunits of the respiratory chain, glycolysis and a chaperone critical to the stabilization and import of proteins. Oxidative products were associated with loss of membrane potential, mitochondrial degeneration and severe axonal loss. Reductions in ATP synthesis were even greater than those associated with mitochondrial diseases. Increasing SOD2 levels by viral mediated gene transfer rescued ATP synthesis, suppressed myelin fiber injury and increased retinal ganglion cell survival 1 year later. PMID- 18421243 TI - First and second language tongue movements in Spanish and Korean bilingual speakers. AB - A number of previous studies have relied on perceptual judgments or acoustic analysis to examine second language (L2) production. However, few researchers have studied L2 performance by directly tracking the physical movements of the articulators. The purpose of the present study was to investigate intraspeaker differences in native (Korean or Spanish) and L2 (English) production through kinematic indices of tongue activity. This involved measurement of the speed, duration, and distance of tongue movements or strokes during speech. Findings indicated that the speakers had significantly slower stroke speeds and longer movement durations for L2 when compared to their native language (L1), yet no significant differences in stroke distance. The bilingual speakers were found to pause more and speak proportionally less of the time in their L2. Interestingly, those speakers who exhibited greater relative kinematic changes from L1 to L2 were also rated as having a stronger perceived accent. PMID- 18421244 TI - 'Frame dominance' and the serial organization of babbling, and first words in Korean-Learning infants. AB - Korean-learning infant patterns in babbling and single words were compared with those of English-learning infants and with Korean adult-directed and infantdirected speech to evaluate the roles of intrinsic production constraints proposed by the frame dominance hypothesis versus early learning mechanisms based on input regularities. Intrasyllabic patterns in babbling of Korean-learning infants were like those of English-learning infants. These patterns were not present in Korean infant-directed speech, providing evidence for the assertion that they are intrinsic to infants, and not triggered by input. Unlike English learning infants, however, Korean-learning infants did not show the expected intrasyllabic patterns in their first words, suggesting that the intrinsic constraints can be overcome in first words if they conflict with ambient speech patterns as they do in Korean. Intersyllabic patterns of Korean-learning infants were mostly similar to those of English-learning infants, showing preferences for consonant manner and vowel height variegation in babbling though only a vowel variegation preference in words. Some implications of the results for generative phonology are considered. PMID- 18421245 TI - Phonetic applications of the time-corrected instantaneous frequency spectrogram. AB - A reassigned or time-corrected instantaneous frequency spectrogram has been developed in the work of a number of practitioners. Here we present a general description of this imaging technique and explore its manifold applications to acoustic phonetics. The TCIF spectrogram shows the locations of signal components with unrivalled precision, eliminating the blurring and smearing of components that hamper the readability of the conventional spectrogram. Formants of vowels and other resonants are shown with great accuracy by observing glottal pulsations at very short time scales with a wideband analysis. A further post-processing technique is also described, by which signal components such as formants, as well as impulsive events, can be effectively isolated to the exclusion of other signal information. When the phonation process is examined this closely, a variety of evidence surfaces which supports recent developments in the theory and computational simulation of aeroacoustic phenomena in speech. Narrowband analysis is also demonstrated to permit pitch tracking with relative ease. PMID- 18421247 TI - Does dopamine still have a leading role in advanced Parkinson's disease after subthalamic stimulation? AB - The role of endogenous dopamine in severe Parkinson's disease is often underestimated. We report on a case of acute general motor worsening induced by the ingestion of fluphenazine in a parkinsonian patient successfully treated with STN DBS. Other etiologies were ruled out. Clinical improvement was gradual and fully reversible 4 days after discontinuation of the antidopaminergic drug. We suggest that residual striatal and extrastriatal dopaminergic pathways still play a paramount role in mediating central neurotrasmissions that may take part in STN DBS's mechanism of action. PMID- 18421248 TI - Check the 'active ingredients' of your medications. Comment on: "Does dopamine still have a leading role in advanced Parkinson's disease after subthalamic stimulation?" (Stereotact Funct Neurosurg 2008;86: 184-186). PMID- 18421249 TI - DBS and dopamine. Comment on "Does dopamine still have a leading role in advanced Parkinson's disease after subthalamic stimulation?" (Stereotact Funct Neurosurg 2008;86: 184-186). PMID- 18421250 TI - A comprehensive review of radiosurgery for cerebral arteriovenous malformations: outcomes, predictive factors, and grading scales. AB - The management of cerebral arteriovenous malformations (AVMs) continues to present a challenge to neurosurgeons. The natural history of this condition, as well as the morbidity and mortality of therapeutic interventions, remains incompletely elucidated. Predictive factors and grading scales in AVM management allow risk-benefit analysis of treatment options and comparison of outcomes. Stereotactic radiosurgery is one of the established treatment modalities for AVMs and is generally used to treat lesions that are high risk for surgical resection. Radiosurgery aims to obliterate AVMs and thus prevent hemorrhage or seizure without any new or worsening of existing symptoms. Lesion characteristics and postsurgical complications differ markedly in patientstreated by radiosurgery versus microsurgery. Radiosurgery-based grading systems account for factors that have been associated with various aspects of radiosurgical outcomes including obliteration, hemorrhage, and postoperative complications, particularly those induced by radiation. The purpose of this paper is to describe the most current predictive factors and grading systems for radiosurgical treatment of cerebral AVMs. PMID- 18421251 TI - Cancer stem cells and survival pathways. AB - Gliomas and medulloblastomas are the most frequent malignant brain tumors in adult and children respectively. Although both tumors arise in the CNS, there is a significant difference in their therapeutic response. Medulloblastomas are relatively curable, while glioblastomas are basically incurable. During the last decade several reports have demonstrated the existence of cancer stem cells in brain tumors, their location and their response to treatment. We have recently described the therapeutic response of medulloblastomas to radiation in their native microenvironment, illustrating how p53 and Pi3K signaling pathways lead to the evasion of cell death by the nestin-expressing cells in the perivascular stem cell niche, even while the bulk of tumor succumbs to apoptosis.(1) It remains to be determined whether this mechanism of tumor resistance applies to the more complex stem-cell niche and tumor bulk of gliomas. PMID- 18421252 TI - Vitamin D for the treatment and prevention of pancreatic cancer. PMID- 18421253 TI - Progress towards the use of HIV protease inhibitors in cancer therapy. AB - Approaches to targeting the signaling pathways responsible for tumor survival are currently under intense investigation and there is ongoing rapid development of novel agents to target various signaling components that mediate tumor growth and survival. The development of agents as viable clinical interventions is, however, a relatively slow process when starting from novel compounds not previously tested in humans. An approach that could circumvent this delay in the development of molecular targeting agents for cancer therapy is to adopt drugs currently in clinical use for other diseases that have the desired effects on cell signaling and to test these for anti-cancer effects. This process is currently being followed with anti-retroviral drugs that inhibit AKT activation as candidates for the treatment of cancer in general, and is being pursued in particular for developing these agent as new means of sensitizing tumors to radiation.(1,2) Although such drugs have been approved for patient use, it is unclear whether or not these drugs can be safely combined with cytotoxic modalities such as radiation. It is this point that Plasteras et al. address in the current issue of Cancer Biology & Therapy, by examining the effects of combined treatment with radiation and HIV protease inhibitors.(3). PMID- 18421254 TI - Role of reactive oxygen species (ROS) and JNKs in selenite-induced apoptosis in HepG2 cells. AB - Dietary selenium (Se) supplementation has been shown to be effective against reducing the risk of incidence of different human cancers. Selenium exists in both organic and inorganic forms. Different chemical forms of selenium metabolize differently in vivo, activate distinct molecular mechanisms and exhibit varying degree of anti-carcinogenicity in different cancer types. The effectiveness of a Se compound could also vary depending on the genetic background of the tumor cells. Therefore, understanding the molecular mechanism(s) by which different Se compounds exert their anti-tumorigenic effects is necessary for their use in cancer chemoprevention. PMID- 18421255 TI - Effects of conserved D/T loop substitutions in the pre-tRNA substrate on tRNase Z catalysis. AB - tRNAs are transcribed as precursors with a 5' end leader and a 3' end trailer. In the course of tRNA maturation, RNase P removes the 5' end leader and tRNase Z can endonucleolytically remove the 3' end trailer. A domain remote from the active site of tRNase Z recognizes and binds substrate, principally through contacts with the elbow (D/T loops) of the tRNA. To evaluate possible contacts, processing kinetics was performed using human nuclear encoded pre-tRNA(Arg) with substitutions in conserved D and T loop nucleotides. Changes in K(M) observed with some of the substitutions suggest contacts between tRNase Z and substrate tRNA in this region, and changes in tRNA structure provide an additional basis for interpretation of the kinetic effects. PMID- 18421256 TI - Interactions between the double-stranded RNA-binding proteins TRBP and PACT define the Medipal domain that mediates protein-protein interactions. AB - The double-stranded (ds) RNA binding proteins, TRBP and PACT bind the interferon induced protein kinase PKR and dsRNA. TRBP inhibits, whereas PACT activates PKR. They have two dsRNA binding domains (dsRBDs) and a C-terminal domain that does not bind RNA. All three domains show a strong homology between the two proteins. Interaction assays by in vitro binding, yeast two-hybrid, and immunoprecipitations show that TRBP and PACT form heterodimers in the absence of dsRNA. In cells, TRBP and PACT colocalize in specific dots of the perinuclear space. Analysis of the individual domains shows that the two dsRBDs of each protein interact with each other. In contrast, the C-terminal domain of PACT homodimerizes and interacts with its homologous region in TRBP, but the same domain in TRBP does not homodimerize. Because the C-terminal domain in TRBP binds to the tumor suppressor Merlin, the RNase III Dicer and PACT, we name it the Merlin Dicer PACT liaison (Medipal) domain. Based on known interactions Medipal is defined as aminoacids 228-366 in TRBP and 195-313 in PACT. TRBP-PACT interaction correlates with an absence of eIF2alpha activation by PACT, suggesting that the heterodimer does not activate PKR. We propose that the Medipal domain mediates specialized functions through protein-protein interactions and contributes to the RNA interference pathway and to PKR activation. PMID- 18421257 TI - Non-invasive ventilation. AB - Non-invasive ventilation (NIV) is an effective technique that can avert side effects and complications associated with endotracheal intubation. NIV is primarily used to avert the need for endotracheal intubation in patients with early stage acute respiratory failure (ARF), and to prevent postextubation respiratory failure in patients considered to be at risk. It can also be used as an alternative to invasive ventilation at a more advanced stage of ARF or to facilitate the process of weaning from mechanical ventilation. The success of NIV relies on several factors including the type and severity of ARF, the underlying disease, the timing, the location of treatment, and the experience of the team. In this review article, we analyze, compare, and discuss the results of studies in which NIV was applied in different pathologies and with different timing during the evolution of ARF. PMID- 18421258 TI - Selection of bacterial populations in the mycosphere of Laccaria proxima: is type III secretion involved? AB - The bacterial communities in the Laccaria proxima mycosphere (soil from beneath fruiting bodies) and the corresponding bulk soil were compared by cultivation dependent and cultivation-independent methods. To assess the distribution of type III secretion systems (TTSS), a PCR-based system for the broad detection of a highly conserved gene involved in TTSS, that is hrcR, was developed and used to assay the cultured bacteria from the L. proxima mycosphere and surrounding bulk soil. PCR-DGGE based on the 16S ribosomal RNA gene showed the selection of presumably mycosphere-specific bacterial groups in the mycosphere of L. proxima compared to the bulk soil in 3 sampling years. Moreover, plate counts revealed that the numbers of culturable heterotrophic bacteria were increased in the mycosphere as compared to the bulk soil. Strikingly, the percentage of randomly picked isolates that carried the hrcR gene showed a significant increase, from 2.8 in the bulk to 13.4 in the mycosphere soil. The increase could be mainly attributed to the emergence of a hrcR positive Pseudomonas fluorescens, denoted BS053, which constituted the most dominant species in the culturable mycosphere communities. This organism was, together with a hrcR-positive Burkholderia terrae BS110, exclusively found in mycosphere soil. Direct detection of hrcR genes using a cultivation-independent approach showed the selection of several hrcR gene types uniquely in the mycosphere, indicating the selection of several TTSS harboring bacterial species. Thus, different bacteria were found to be enriched in the L. proxima mycosphere and TTSS can be involved in some of the interactions with the fungal host. PMID- 18421259 TI - Humoral immunity to commensal oral bacteria in human infants: evidence that Streptococcus mitis biovar 1 colonization induces strain-specific salivary immunoglobulin A antibodies. AB - To define the relationship between salivary SIgA antibodies and commensal oral bacteria, we examined the reactivity of SIgA antibodies from the saliva of four infants with their own colonizing Streptococcus mitis biovar 1 (S. mitis bv 1) clones (ribotypes). Immunoblot analysis was used to examine reactivity of these antibodies with persistent ribotypes isolated from the mouths of the infants over the first year postpartum. Results showed that the pattern of SIgA antibody reactivity with the majority of clones increased in complexity after colonization but that most additional bands were common to other clones, indicating that they represented shared antigens. However, unique bands were identified in 75% of the selected persistent clones. We hypothesized that if strain-specific SIgA antibody was induced in response to colonization of a particular clone and contributed to its elimination from the mouth, then the appearance of unique bands would immediately precede the disappearance of the strain. Seventy-three percent of all unique bands identified in the study fulfilled this criterion. Because the mouth is an open, dynamic environment and multiple factors are believed to play a role in the immune response at mucosal surfaces, it may not be possible to conclusively define the relationship between SIgA antibody and commensal bacteria. However, our data provide evidence that SIgA antibody, reactive with unique antigens of their own colonizing strains, is produced in infants and may point to a role of this antibody in regulating colonization by S. mitis bv 1. PMID- 18421260 TI - BMI compared with central obesity indicators in relation to diabetes and hypertension in Asians. AB - OBJECTIVE: To compare BMI with waist circumference (WC), waist-to-hip ratio (WHR), and waist-to-stature ratio (WSR) in association with diabetes or hypertension. METHODS AND PROCEDURES: Cross-sectional data from 16 cohorts from the DECODA (Diabetes Epidemiology: Collaborative Analysis of Diagnostic criteria in Asia) study, comprising 9,095 men and 11,732 women, aged 35-74 years, of different ethnicities were included in this meta-analysis. RESULTS: Age-adjusted odds ratios (ORs) for diabetes in men (women) for 1 s.d. increase in BMI, WC, WHR, and WSR were 1.52 (1.59), 1.54 (1.70), 1.53 (1.50), and 1.62 (1.70), respectively; and the corresponding ORs for hypertension were 1.68 (1.55), 1.66 (1.51), 1.45 (1.28), and 1.63 (1.50). Paired homogeneity tests (BMI with each of the three) adjusted for age and cohort showed that diabetes had stronger association with WSR than BMI (P=0.001) in men but with WC and WSR than BMI (both P<0.05) in women. Hypertension had stronger association with BMI than WHR in men (P<0.001) and had the strongest with BMI than the others (WHR P<0.001; WSR P<0.01; and WC P<0.05) in women. Areas under the receiver operating characteristic (ROC) curves adjusted for age and cohort were slightly larger for diabetes for WSR 0.735 (0.748) in men (women) and WC 0.749 (women only) than BMI 0.725 (0.742) while for hypertension larger for BMI 0.760 (0.766) than WHR 0.748 (0.751), but their 95% CIs were all overlapped. DISCUSSION: WSR was stronger than BMI in association with diabetes, but these indicators were equally strongly associated with hypertension in Asians. PMID- 18421261 TI - Estradiol and the estradiol metabolite, 2-hydroxyestradiol, activate AMP activated protein kinase in C2C12 myotubes. AB - OBJECTIVE: Systemic loss of estradiol (E2) during menopause is associated with increased adiposity which can be prevented with E2 replacement. Rodent studies suggest that E2, or lack of, is a key mediator in menopause-related metabolic changes. We have previously demonstrated that E2 treatment produces a rapid, dose dependent activation of AMP-activated protein kinase (AMPK) in murine skeletal muscle. Activation of AMPK is implicated in the therapeutic benefits of many insulin sensitizing agents including metformin and thiazolidinediones. Here, we expand our observations and provide novel data which demonstrate that in addition to E2, its metabolite 2-hydroxyestradiol (2-HE2), activate AMPK in C2C12 myotubes. METHODS AND PROCEDURES: C2C12 myotubes were used to examine the effects on E2 and the by-products of its metabolism on AMPK activation. RESULTS: Low concentrations of E2 (10 and 100 nmol/l) were found to increase AMPK phosphorylation by approximately 1.6-fold, while a higher concentration (10 micromol/l) resulted in a approximately 3.0-fold increase. In comparison to E2 treatment alone, incubation of myotubes with E2 and 1-aminobenzotriazole (ABT) (a CYP450 inhibitor that blocks metabolism of E2) caused AMPK activation to be enhanced at low E2 concentrations, but attenuated at higher concentrations. The effects of ABT suggested that one or more E2 metabolites contribute to the maximal activation of AMPK at high E2 concentrations. Indeed, the estrogen metabolite 2-HE2, but not 2-methoxyestradiol (2-ME2), directly activated AMPK in C2C12 myotubes. DISCUSSION: We propose a model where E2, acting through its metabolite 2-HE2 and the estrogen receptors (ERs), activates AMPK in myotubes. Finally, activation is abolished when all E2 is metabolized to 2-ME2. PMID- 18421262 TI - Childhood obesity: genetic and environmental overlap with normal-range BMI. AB - OBJECTIVE: To understand the overlap between the etiology of obesity and normal variation in BMI in children. METHODS AND PROCEDURES: Height and weight data were available from a large UK representative sample of twins: 2,342 same-sex pairs at 7 years and 3,526 same-sex pairs at 10 years. The twin method and model-fitting techniques were used to estimate genetic and environmental contributions to BMI. DeFries-Fulker (DF) extremes analysis was used to investigate genetic and environmental influences on the mean difference between obese and normal-weight children. Obesity was classified using the International Obesity Task Force (IOTF) criteria. RESULTS: At both ages, BMI and obesity were highly heritable (0.60-0.74) and only modestly influenced by shared environmental factors (0.12 0.22). Extremes analyses indicated that genetic and environmental influences on obesity are quantitatively and qualitatively similar to those operating across the range of BMI. DISCUSSION: Obesity is the extreme of the same genetic and environmental factors responsible for variation throughout the distribution of BMI. This finding implies that genes that influence obesity will also be associated with BMI in the normal range, and similar environmental influences will affect BMI in the clinical and normal range. Knowing that obesity is influenced by the same genetic and environmental factors that affect weight at all levels has implications for investigating the mechanisms for weight gain and developing interventions for weight control. PMID- 18421263 TI - Maternal and paternal controlling feeding practices: reliability and relationships with BMI. AB - OBJECTIVE: This study aimed to examine the interrelationships between mothers' and fathers' reports on the child-feeding questionnaire (CFQ), the BMI of parents and their children, and observations of parents' controlling feeding practices at mealtimes. METHODS AND PROCEDURES: Twenty-three mothers and twenty-three fathers of children aged between 18 and 67 months reported on their child-feeding practices, on their child's height and weight, and were observed during a normal family mealtime at home. RESULTS: No associations were found between mothers' reported and observed feeding practices. Fathers' reported pressure to eat and restriction were associated with more controlling observed mealtime feeding practices. Mothers and fathers did not significantly differ in their reported or observed child-feeding practices. Children's BMI was not related to maternal or paternal reported or observed feeding practices. More mealtime pressure was observed in parents with a higher BMI. DISCUSSION: Fathers' self-reports of their mealtime practices are reliable. Mothers' feeding practices may differ when fathers are present and further work should examine mothers at mealtimes with and without fathers. Although children's BMI was not related to parents' use of reported or observed control, parents with a higher BMI were more controlling, highlighting the importance of considering parents' own weight in future studies. PMID- 18421264 TI - Relationship between BMI and all-cause mortality in Japan: NIPPON DATA80. AB - As body composition in Asian populations is largely different from Western populations, a healthy BMI could also differ between the two populations. Thus, further study is needed to determine whether a healthy BMI in Asians should be lower than Western populations, as recommended by the World Health Organization (WHO). We investigated the relationship between BMI and mortality in a sample of 8,924 Japanese men and women without stroke or heart disease. During 19 years of follow-up, 1,718 deaths were observed. We found a U-shaped relationship between BMI and fatal events. Risk of total mortality was highest in participants with BMI <18.5 kg/m(2) and lowest in participants with BMI 23.0-24.9 kg/m(2). These findings persisted even after excluding the first 5 years of follow-up with a focus on healthy participants who never smoked, were aged <70 years, and had total cholesterol (TC) levels >or=4.1 mmol/l (N=3712). For both the full sample and healthy participants, all-cause mortality risk did not differ between BMI ranges 21.0-22.9 and 23.0-24.9 kg/m(2). Our findings do not support the recent WHO implications that BMIs <23.0 kg/m(2) is healthy for Asians. Therefore, further studies are needed to identify an optimal BMI range for Asia. PMID- 18421265 TI - Genome-wide linkage scan for the metabolic syndrome: the GENNID study. AB - In the United States, the metabolic syndrome (MetS) constitutes a major public health problem with over 47 million persons meeting clinical criteria for MetS. Numerous studies have suggested genetic susceptibility to MetS. The goals of this study were (i) to identify susceptibility loci for MetS in well-characterized families with type 2 diabetes (T2D) in four ethnic groups and (ii) to determine whether evidence for linkage varies across the four groups. The GENNID study (Genetics of NIDDM) is a multicenter study established by the American Diabetes Association in 1993 and comprises a comprehensive, well-characterized resource of T2D families from four ethnic groups (whites, Mexican Americans, African Americans, and Japanese Americans). Principal component factor analysis (PCFA) was used to define quantitative phenotypes of the MetS. Variance components linkage analysis was conducted using microsatellite markers from a 10-cM genome wide linkage scan, separately in each of the four ethnic groups. Three quantitative MetS factors were identified by PCFA and used as phenotypes for MetS: (i) a weight/waist factor, (ii) a blood pressure factor, and (iii) a lipid factor. Evidence for linkage to each of these factors was observed. For each ethnic group, our results suggest that several regions harbor susceptibility genes for the MetS. The strongest evidence for linkage for MetS phenotypes was observed on chromosome 2 (2q12.1-2q13) in the white sample and on chromosome 3 (3q26.1-3q29) in the Mexican-American sample. In conclusion, the results suggest that several regions harbor MetS susceptibility genes and that heterogeneity may exist across groups. PMID- 18421266 TI - Body fat and mobility are explained by common genetic and environmental influences in older women. AB - In older adults, mobility limitations often coexist with overweight or obesity, suggesting that similar factors may underlie both traits. This study examined the extent to which genetic and environmental influences explain the association between adiposity and mobility in older women. Body fat percentage (bioimpedance test), walking speed over 10 m, and distance walked in a 6-min test were evaluated in 92 monozygotic (MZ) and 104 dizygotic (DZ) pairs of twin sisters reared together, aged 63-76 years. Genetic and environmental influences on each trait were estimated using age-adjusted multivariate genetic modeling. The analyses showed that the means (and s.d.) for body fat percentage, walking speed, and walking endurance were 33.2+/-7.3%, 1.7+/-0.3 m/s and 529.7+/-75.4 m, respectively. The phenotypic correlation between adiposity and walking speed was 0.32 and between adiposity and endurance it was -0.33. Genetic influences explained 80% of the association between adiposity and speed, and 65% of adiposity and walking endurance. Cross-trait genetic influences accounted for 12% of the variability in adiposity, 56% in walking speed, and 34% in endurance. Trait-specific genetic influences were also detected for adiposity (54%) and walking endurance (13%), but not speed. In conclusion, among community-living older women, an inverse association was found between adiposity and mobility that was mostly due to the effect of shared genes. This result suggests that the identification of genetic variants for body fat metabolism may also provide understanding of the development of mobility limitations in older women. PMID- 18421267 TI - Does high BMI influence hospital charges in children undergoing adenotonsillectomy? AB - BACKGROUND: Obesity is a highly prevalent chronic problem with health and fiscal consequences. Data from adults and nonsurgical pediatric patients suggest that obesity has serious implications for the US economy. OBJECTIVE: Our goal was to describe the impact of BMI on hospital charges in children undergoing adenotonsillectomy (AT). METHODS AND PROCEDURES: We carried out a retrospective comparative analysis of the electronic anesthesia record and the charges from billing data from a large tertiary institution on children aged 3-18 years who had AT during the year 2005-2007. The main outcome measures were mean total hospital charges, likelihood of admission, and length of hospital stay (LOS). RESULTS: Of 1,643 children, 68.9% were aged <10 years, 76% were whites, and 74.1% had private commercial insurance. Most (75.3%) children were discharged on the day of surgery. Obese and overweight children were more likely to be admitted than their normal-weight peers (X(2)=26.3, P<0.001). Among those admitted, BMI showed a positive correlation with LOS (r=0.20, P<0.001). Obese and overweight patients had significantly higher total hospital charges than their healthy weight counterparts (P=0.001). Anesthesia, postanesthesia care unit (PACU), and pharmacy and laboratory charges were also higher for obese than normal-weight children (P<0.05). DISCUSSION: Overweight and obese children undergoing AT accrued higher hospital charges and had longer postoperative LOS than their healthy-weight peers. If these findings are extendable to other surgical procedures, they could have far-reaching implications for the US economy. PMID- 18421268 TI - Determinants of insulin-resistant phenotypes in normal-weight and obese Black African women. AB - OBJECTIVE: Subsets of metabolically "healthy obese" and "at-risk" normal-weight individuals have been previously identified. The aim of this study was to explore the determinants of these phenotypes in black South African (SA) women. METHODS AND PROCEDURES: From a total of 103 normal-weight (BMI or= 30 kg/m(2)) black SA women, body composition, fat distribution, blood pressure, fasting glucose levels, insulin resistance, and lipid profiles were measured. Questionnaires relating to family history, physical activity energy expenditure (PAEE), and socio-demographic variables were administered. The subjects were classified as insulin sensitive or insulin resistant according to the homeostasis model assessment of insulin resistance (HOMA-IR) (>or=1.95 insulin resistant). RESULTS: Our study showed that 22% of the normal-weight women were insulin resistant and 38% of the obese women were insulin sensitive. Increased visceral adipose tissue (VAT) (P=0.001) and decreased VAT/leg fat mass (P 30 kg/m(2)) male and female subject volunteers with normal oral glucose tolerance test. Saline or GLP-1m were infused from 0 to 60 min in each study. Additionally, seven lean and six obese subjects underwent a third clamp in which the GLP-1 receptor (GLP-1R) antagonist, exendin (9-39) amide was infused from -60 to 60 min with GLP-1m from 0 to 60 min. RESULTS: No glucose infusion was required in lean subjects to sustain euglycemia (glucose clamp) during saline or GLP-1m infusions. However, in obese subjects glucose infusion was necessary during GLP 1m infusion alone in order to compensate for a marked (>50%) inhibition of hepatic glucose production (HGP). Plasma insulin levels remained constant in lean subjects but rose significantly in obese subjects after termination of the peptide infusions. During GLP-1R blockade, infusion of glucose was immediately required upon starting GLP-1m infusions in all subjects due to a more dramatic reduction in HGP, as well as a delayed and modest insulinotropic response. DISCUSSION: We conclude that GLP-1m potently inhibits HGP and is a weak insulinotropic agent. These properties are particularly apparent and pronounced in obese but only become apparent in lean subjects during GLP-1 (7-36) receptor blockade. These previously unrecognized antidiabetogenic actions of GLP-1m may have therapeutic usefulness. PMID- 18421271 TI - State ranks of incident cancer burden due to overweight and obesity in the United States, 2003. AB - OBJECTIVE: Given links between obesity and cancer, we estimated incident cancer burden due to overweight and obesity at the state level in the United States. METHODS AND PROCEDURES: Using state rankings by per capita burden of incident cancer cases diagnosed in 2003 that were related to overweight and obesity, we examined the frequency with which states ranked in the highest and lowest quintiles of weight-related burden for cancers of the postmenopausal breast, endometrium, kidney, colon, and prostate. In this study, data from the Behavioral Risk Factor Surveillance System (BRFSS), US Census, US Mortality Public Use Data Tapes, and National Cancer Institute Surveillance, Epidemiology, and End Results (SEER) Program were used. RESULTS: Western states had the lowest weight-related cancer burden for both sexes. Iowa, South Dakota, and West Virginia had the highest burden for all three types of male cancers. West Virginia is the only state that ranked in the quintile of highest weight-related burden for all four cancers considered in women. DISCUSSION: For certain cancers, including endometrial, postmenopausal breast, and colon cancers, states with high burdens clustered in geographic regions, warranting further inquiry. Although state ranks for the total cancer burden and the prevalence of overweight and obesity correlated with state ranks for weight-related incident cancer burden, they often served poorly as its proxy. Such a finding cautions against simply targeting states with high overweight and obesity or high total burdens of cancers for which overweight and obesity are risk factors, as this approach may not reach areas of unrecognized burden. PMID- 18421272 TI - Ability to taste 6-n-propylthiouracil and BMI in low-income preschool-aged children. AB - BACKGROUND: Sensitivity to the bitter compound 6-n-propylthiouracil (PROP) is genetically mediated. Sensitivity to PROP has been associated with weight status in both adults and children. OBJECTIVE: To determine whether there is an association between PROP sensitivity and BMI in low-income children of diverse race/ethnicity, among whom there is a high prevalence of obesity. METHODS AND PROCEDURES: Eighty-one preschool-aged children attending Head Start tasted a solution of 560 micromol/l PROP and reported whether it tasted "like water" or "like something else". Mothers reported child's race, age, maternal education, maternal weight and height, child's reluctance to sample new foods via the Food Neophobia Scale (FNS), and child's dietary intake using a food frequency questionnaire. Child weight and height were measured. BMI was calculated and for children, expressed in z-scores. Regression analyses were used to evaluate the relationship between child's PROP taster status and BMI z-score, testing covariates child's age, gender, race, maternal education and BMI, and child's FNS score. Children's dietary intake was compared by PROP taster status. RESULTS: PROP tasters, compared with nontasters, had significantly higher BMI z-scores (0.99 (s.d. 1.24) vs. 0.03 (1.12), P=0.004) and had a significantly higher prevalence of overweight (31.8% vs. 5.6%, P=0.025), but demonstrated no differences in reported dietary intake. The most parsimonious model predicting the child's BMI z-score included only maternal BMI and the child's PROP taster status (R(2)=22.3%). DISCUSSION: A genetically mediated ability to taste bitter may contribute to obesity risk in low-income, preschool-aged children. PMID- 18421273 TI - Weight loss of black, white, and Hispanic men and women in the Diabetes Prevention Program. AB - OBJECTIVE: To provide the specific weight loss outcomes for African-American, Hispanic, and white men and women in the lifestyle and metformin treatment arms of the Diabetes Prevention Program (DPP) by race-gender group to facilitate researchers translating similar interventions to minority populations, as well as provide realistic weight loss expectations for clinicians. METHODS AND PROCEDURES: Secondary analyses of weight loss of 2,921 overweight participants (22% black; 17% Hispanic; 61% white; and 68% women) with impaired glucose tolerance randomized in the DPP to intensive lifestyle modification, metformin or placebo. Data over a 30-month period are examined for comparability across treatment arms by race and gender. RESULTS: Within lifestyle treatment, all race gender groups lost comparable amounts of weight with the exception of black women who exhibited significantly smaller weight losses (P < 0.01). For example, at 12 months, weight losses for white men (-8.4%), white women (-8.1%), Hispanic men ( 7.8%), Hispanic women (-7.1%), and black men (-7.1%) were similar and significantly higher than black women (-4.5%). In contrast, within metformin treatment, all race-gender groups including black women lost similar amounts of weight. Race-gender specific mean weight loss data are provided by treatment arm for each follow-up period. DISCUSSION: Diminished weight losses were apparent among black women in comparison with other race-gender groups in a lifestyle intervention but not metformin, underscoring the critical nature of examining sociocultural and environmental contributors to successful lifestyle intervention for black women. PMID- 18421274 TI - Effects of a novel Y5 antagonist in obese mice: combination with food restriction or sibutramine. AB - OBJECTIVE: To further address the function of the Y5 receptor in energy homeostasis, we investigated the effects of a novel spironolactone Y5 antagonist in diet-induced obese (DIO) mice. METHODS AND PROCEDURES: Male C57BL/6 or Npy5r( /-) mice were adapted to high-fat (HF) diet for 6-10 months and were submitted to three experimental treatments. First, the Y5 antagonist at a dose of 10 or 30 mg/kg was administered for 1 month to DIO C57BL/6 or Npy5r(-/-) mice. Second, the Y5 antagonist at 30 mg/kg was administered for 1.5 months to DIO C57BL/6 mice, and insulin sensitivity was evaluated using an insulin tolerance test. After a recovery period, nuclear magnetic resonance measurement was performed to evaluate body composition. Third, DIO mice were treated with the Y5 antagonist alone, or in combination with 10% food restriction, or with another anorectic agent, sibutramine at 10 mg/kg, for 1.5 months. Plasma glucose, insulin, and leptin levels, and adipose tissue weights were quantified. RESULTS: The spironolactone Y5 antagonist significantly reduced body weight in C57BL DIO mice, but not in Npy5r(-/-) DIO mice. The Y5 antagonist produced a fat-selective loss of body weight, and ameliorated obesity-associated insulin resistance in DIO mice. In addition, the Y5 antagonist combined with either food restriction or sibutramine tended to produce greater body weight loss, as compared with single treatment. DISCUSSION: These findings demonstrate that the Y5 receptor is an important mediator of energy homeostasis in rodents. PMID- 18421275 TI - Stress and sex influences on food-seeking behaviors. AB - OBJECTIVE: Stress increases the drive to consume calorically dense preferred foods suggesting an exogenous factor that may induce caloric overconsumption and weight gain. As females show heightened stress sensitivity and present with increased rates of obesity, we hypothesized that stress-induced increases in the motivation for preferred foods may be a sex-specific predisposing factor for weight gain. METHODS AND PROCEDURES: To investigate this hypothesis, we have developed a buried food paradigm that permits the measurement of sex differences and effects of chronic variable stress (CVS) on the latency to uncover and the consumption of a preferred food pellet without the requisite caloric restriction required in traditional operant conditioning tasks. RESULTS: In our studies, females consistently showed latencies that were twice as fast as males to locate the buried pellet in limited access tests. Interestingly, during stress exposure, male latencies decreased to that of control female levels. Male and female mice showed a significant effect of stress, three- and fourfold, respectively, on increased consumption of the preferred food during testing. DISCUSSION: These results support a basal sex difference in behaviors toward a preferred food, and a possible role of stress sensitivity in the drive and intake of such foods. Sex differences in the role stress plays in these behaviors may provide insight into underlying mechanisms related to an increased obesity risk. PMID- 18421276 TI - 11beta-hydroxysteroid dehydrogenase 2 activity is elevated in severe obesity and negatively associated with insulin sensitivity. AB - Alterations in glucocorticoid (GC) metabolism may contribute to the development of obesity and insulin resistance. We aimed to study the role of 11beta hydroxysteroid dehydrogenase type 2 (11beta-HSD2) in human adiposity, paying special attention to the association between altered GC metabolism and insulin sensitivity. In 24-h urine samples of 72 extremely obese (mean BMI 45.5 +/- 1.1 kg/m(2)), but otherwise healthy patients urinary free cortisol (UFF), urinary free cortisone (UFE), tetrahydrocortisol (THF), 5alpha-tetrahydrocortisol (5alpha THF), and tetrahydrocortisone (THE) were quantified by radioimmunoassay. The sum of the three major tetrahydrometabolites is an estimate for daily GC secretion, and the sum of UFF and UFE represents potentially bioactive-free-GCs. Thirty healthy lean subjects (BMI 22.3 +/- 0.3 kg/m(2)) served as controls. In obese subjects, absolute daily GC secretion and the potentially bioactive-free-GCs were significantly (P < 0.005) higher than in lean controls (11.8 +/- 0.7 vs. 8.0 +/- 0.6 mg/d; and 171.8 +/- 11.2 vs. 117.6 +/- 9.2 mug/d, respectively). However, when these values were corrected for body surface area (BSA), significant differences were no longer detectable. While enzyme activity indices for 5alpha reductase and 11beta-hydroxysteroid dehydrogenase type 1 (11beta-HSD1) were similar in lean and obese subjects, 11beta-HSD2 was markedly elevated in adiposity (3.7 +/- 0.2 vs. 2.1 +/- 0.1; P < 0.0001). This increase was accompanied by a significant reduction in UFF excretion corrected for BSA (16.5 +/- 1.2 vs. 21.7 +/- 2.0 mug/d/m(2); P = 0.0222). Besides, 11beta-HSD2 activity was significantly correlated with insulin sensitivity (P = 0.0262). When body size is accounted for, both adrenal GC secretion and potentially bioactive-free GCs are indistinguishable between lean and extremely obese subjects. However in obesity, the kidney appears to intensify its supply of the direct substrate cortisone for extrarenal 11beta-HSD1, which may fuel visceral adiposity and insulin resistance. PMID- 18421277 TI - Physical activity and healthy weight maintenance from childhood to adulthood. AB - The objective of this study was to determine whether change in physical activity was associated with maintaining a healthy weight from childhood to adulthood. This prospective cohort study examined 1,594 young Australian adults (48.9% female) aged 27-36 years who were first examined at age 9-15 years as part of a national health and fitness survey. BMI was calculated from measured height and weight, and physical activity was self-reported at both time points; pedometers were also used at follow-up. Change in physical activity was characterized by calculating the difference between baseline and follow-up z-scores. Change scores were categorized as decreasing (large, moderate), stable, or increasing (large, moderate). Healthy weight was defined in childhood as a BMI less than international overweight cutoff points, and in adulthood as BMI<25 kg/m(2). Healthy weight maintainers were healthy weight at both time points. Compared with those who demonstrated large relative decreases in physical activity, females in all other groups were 25-37% more likely to be healthy weight maintainers, although associations differed according to the physical activity measure used at follow-up and few reached statistical significance. Although younger males whose relative physical activity moderately or largely increased were 27-34% more likely to be healthy weight maintainers than those whose relative physical activity largely decreased, differences were not statistically significant. In conclusion, relatively increasing and stable physical activity from childhood to adulthood was only weakly associated with healthy weight maintenance. Examining personal, social, and environmental factors associated with healthy weight maintenance will be an important next step in understanding why some groups avoid becoming overweight. PMID- 18421278 TI - Contribution of Midparental BMI and other determinants of obesity in adult offspring. AB - The aim of this study was to evaluate midparental BMI among intergenerational factors associated with obesity in adult offspring. The data was from an unusual two-generational observational design of 1,477 married couples from Renfrew and Paisley in Scotland who were aged 45-64 years when screened in 1972-1976, and 1,040 sons and 1,298 daughters aged 30-59 years when screened in 1996. BMI was categorized as normal (< 25 kg/m(2)), overweight (25-29.9 kg/m(2)), and obese (> or = 30 kg/m(2)) in offspring and parents. Midparental BMI was defined as the mean of the mother's and father's BMI. Low physical activity, nonsmoking status, higher cholesterol level, and manual social class were all associated with increased BMI in offspring. The effect of reported dietary intake was less clear. Offspring of obese parents (defined by midparental BMI) were over four times more likely to be obese than offspring of normal weight parents. Midparental BMI had a strong effect on offspring BMI, independent of social class, smoking habit, physical activity, and reported dietary intake. Adding midparental BMI to the regression model more than doubled the explained variation of offspring BMI from 7.7 to 17%. Every 1 kg/m(2) increment in midparental BMI was associated with a BMI greater by 0.51 kg/m(2) in offspring. We conclude that midparental BMI is a useful simple tool to predict offspring BMI. Whether it represents genetic or environmental family effects, it is easily ascertained by the individual and could be used in health promotion and clinical settings to target individuals who are at increased risk of becoming obese. PMID- 18421279 TI - Tlr-4 deficiency selectively protects against obesity induced by diets high in saturated fat. AB - Toll-like receptor-4 (Tlr-4), a key pattern recognition receptor involved in innate immune response, is activated by saturated fatty acids (SFAs). To investigate the involvement of this receptor in obesity caused by consumption of diets high in fat, we utilized male Tlr-4-deficient 10ScN mice and 10J controls. Mice were fed either low fat (low-fat control (LFC)), high unsaturated fat (high fat control (HFC)), or high saturated fat + palmitate (HFP) diets ad libitum for 16 weeks. Relative to the LFC diet, the HFC diet resulted in greater epididymal fat pad weights and adipocyte hypertrophy in both Tlr-4-deficient and normal mice. However, the 10ScN mice were completely protected against the obesigenic effects of the HFP diet. Moreover, macrophage infiltration and monocyte chemotactic protein-1 (MCP-1) transcript abundance were lower in adipose tissue of 10ScN mice fed the HFP diet, and the hyperinsulinemic response was negated. Tlr-4-deficient mice also had markedly lower circulating concentrations of MCP-1 and much less nuclear factor-kappaB (NFkappaB) protein in nuclear extracts prepared from adipose tissue, irrespective of diet. In contrast, Tlr-4 deficiency did not attenuate the induction of tumor necrosis factor-alpha (TNF-alpha) or interleukin-6 (IL-6) expression in adipose tissue. These data indicate that Tlr-4 deficiency selectively protects against the obesigenic effects of SFA and alters obesity-related inflammatory responses in adipose tissue. PMID- 18421280 TI - Effects of the new C1q/TNF-related protein (CTRP-3) "cartonectin" on the adipocytic secretion of adipokines. AB - BACKGROUND: Cartonectin (collagenous repeat-containing sequence of 26-kDa protein; CORS-26) was described as a new adipokine of the C1q/TNF molecular superfamily C1q/TNF-related protein-3 (CTRP-3), secreted by the adipocytes of mice and humans. The receptor and function of cartonectin are unknown and the recombinant protein is not commercially available. OBJECTIVE: To investigate the effects of recombinant cartonectin on the secretion of adipokines such as adiponectin, leptin, and resistin from adipocytes of human and murine origin. The effect of the BMI of the adipocyte donor was also investigated. METHODS AND PROCEDURES: Human adipocytes from pooled lean and preobese healthy individuals and murine 3T3-L1 adipocytes were used for stimulation experiments. Recombinant cartonectin was expressed in insect H5 cells. Adipokine secretion was measured using enzyme-linked immunosorbent assay. In addition, western blot analysis and luciferase reporter gene assays were employed. RESULTS: Cartonectin (1, 10, 50, and 250 ng/ml) in higher doses stimulates the secretion of adiponectin and resistin from murine adipocytes. This effect is not caused by an induction of peroxisome proliferator-activated receptor-gamma (PPAR-gamma) protein expression, as confirmed by western blot analysis. Also, luciferase reporter gene assay revealed that cartonectin failed to induce luciferase activity at the peroxisome proliferator-activated receptor responsive element site containing the adiponectin/luciferase promoter fragment. Human adipocytes from lean individuals secrete higher amounts of adiponectin and leptin when compared with adipocytes of individuals with a preobesity BMI (25-30 kg/m(2)). Cartonectin failed to stimulate adiponectin or leptin secretion from human adipocytes, irrespective of the BMI value. DISCUSSION: Cartonectin is a new adipokine that differentially regulates the secretion of classical adipokines, with marked differences between the human and the murine systems. These effects are species-dependent, while basal adipokine secretion is influenced by the BMI. PMID- 18421282 TI - Mannoside hydrolysis: it's a boat! PMID- 18421283 TI - Connecting the dots. PMID- 18421281 TI - Effect of 6-month calorie restriction and exercise on serum and liver lipids and markers of liver function. AB - OBJECTIVE: Nonalcoholic fatty liver disease (NAFLD) and its association with insulin resistance are increasingly recognized as major health burdens. The main objectives of this study were to assess the relation between liver lipid content and serum lipids, markers of liver function and inflammation in healthy overweight subjects, and to determine whether caloric restriction (CR) (which improves insulin resistance) reduces liver lipids in association with these same measures. METHODS AND PROCEDURES: Forty-six white and black overweight men and women (BMI = 24.7-31.3 kg/m(2)) were randomized to "control (CO)" = 100% energy requirements; "CR" = 25%; "caloric restriction and increased structured exercise (CR+EX)"= 12.5% CR + 12.5% increase in energy expenditure through exercise; or "low-calorie diet (LCD)" = 15% weight loss by liquid diet followed by weight maintenance, for 6 months. Liver lipid content was assessed by magnetic resonance spectroscopy (MRS) and computed tomography (CT). Lipid concentrations, markers of liver function (alanine aminotransferase (ALT), alkaline phosphatase (ALK)), and whole-body inflammation (tumor necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6), high-sensitivity C-reactive protein (hsCRP)) were measured in fasting blood. RESULTS: At baseline, increased liver lipid content (by MRS) correlated (P < 0.05) with elevated fasting triglyceride (r = 0.52), ALT (r = 0.42), and hsCRP (r = 0.33) concentrations after adjusting for sex, race, and alcohol consumption. With CR, liver lipid content was significantly lowered by CR, CR+EX, and LCD (detected by MRS only). The reduction in liver lipid content, however, was not significantly correlated with the reduction in triglycerides (r = 0.26; P = 0.11) or with the changes in ALT, high-density lipoprotein (HDL)-cholesterol, or markers of whole-body inflammation. DISCUSSION: CR may be beneficial for reducing liver lipid and lowering triglycerides in overweight subjects without known NAFLD. PMID- 18421285 TI - Beta-mannoside hydrolysis goes by boat. PMID- 18421286 TI - Engineering protease specificity made simple, but not simpler. PMID- 18421287 TI - Encoding synthetic polymers. PMID- 18421288 TI - Do-it-yourself enzymes. PMID- 18421289 TI - Breaking the barriers of translation. PMID- 18421291 TI - Reconciling the chemistry and biology of reactive oxygen species. AB - There is a vast literature on the generation and effects of reactive oxygen species in biological systems, both in relation to damage they cause and their involvement in cell regulatory and signaling pathways. The biological chemistry of different oxidants is becoming well understood, but it is often unclear how this translates into cellular mechanisms where redox changes have been demonstrated. This review addresses this gap. It examines how target selectivity and antioxidant effectiveness vary for different oxidants. Kinetic considerations of reactivity are used to assess likely targets in cells and how reactions might be influenced by restricted diffusion and compartmentalization. It also highlights areas where greater understanding is required on the fate of oxidants generated by cellular NADPH oxidases and on the identification of oxidant sensors in cell signaling. PMID- 18421293 TI - Meta-analysis of two genome-wide association studies of bipolar disorder reveals important points of agreement. PMID- 18421294 TI - Decrease in hippocampal neurogenesis during pregnancy: a link to immunity. PMID- 18421295 TI - IARC evaluation of ELF magnetic fields: public understanding of the 0.4-microT exposure metric. PMID- 18421298 TI - Assembly of the three small Tim proteins precedes docking to the mitochondrial carrier translocase. AB - The mitochondrial intermembrane space contains a family of small Tim proteins that function as essential chaperones for protein import. The soluble Tim9-Tim10 complex transfers hydrophobic precursor proteins through the aqueous intermembrane space to the carrier translocase of the inner membrane (TIM22 complex). Tim12, a peripheral membrane subunit of the TIM22 complex, is thought to recruit a portion of Tim9-Tim10 to the inner membrane. It is not known, however, how Tim12 is assembled. We have identified a new intermediate in the biogenesis pathway of Tim12. A soluble form of Tim12 first assembles with Tim9 and Tim10 to form a Tim12-core complex. Tim12-core then docks onto the membrane integrated subunits of the TIM22 complex to form the holo-translocase. Thus, the function of Tim12 in linking soluble and membrane-integrated subunits of the import machinery involves a sequential assembly mechanism of the translocase through a soluble intermediate complex of the three essential small Tim proteins. PMID- 18421299 TI - Inhibition of SHP2 leads to mesenchymal to epithelial transition in breast cancer cells. AB - The Src homology phosphotyrosyl phosphatase 2 (SHP2) is an essential transducer of mitogenic and cell survival signaling in the epidermal growth factor receptor (EGFR) signaling pathway. However, the role of SHP2 in aberrant EGFR and human EGFR2 (HER2) signaling and cancer, particularly in breast cancer, has not been investigated. Here, we report that SHP2 is required for mitogenic and cell survival signaling and for sustaining the transformation phenotypes of breast cancer cell lines that overexpress EGFR and HER2. Inhibition of SHP2 suppressed EGF-induced activation of the Ras-ERK and the phosphatidylinositol 3 kinase-Akt signaling pathways, abolished anchorage-independent growth, induced epithelial cell morphology and led to reversion to a normal breast epithelial phenotype. Furthermore, inhibition of SHP2 led to upregulation of E-cadherin (epithelial marker) and downregulation of fibronectin and vimentin (mesenchymal markers). These results indicate that SHP2 promotes breast cancer cell phenotypes by positively modulating mitogenic and cell survival signaling, by suppressing E cadherin expression which is known to play a tumor suppressor role and by sustaining the mesenchymal state as evidenced by the positive impact on fibronectin and vimentin expression. Therefore, SHP2 promotes epithelial to mesenchymal transition, whereas its inhibition leads to mesenchymal to epithelial transition. On the basis of these premises, we propose that interference with SHP2 function might help treat breast cancer. PMID- 18421300 TI - Direct activation of caspase 8 by the proapoptotic E2 protein of HPV18 independent of adaptor proteins. AB - The self-activation of initiator caspases is dependent on their oligomerization driven by interaction with the death fold domains (DFD) of adaptor proteins. Here, we show that the E2 protein of human papillomavirus type 18 triggers apoptosis by assembling cytoplasmic filaments together with caspase 8, in which its efficient self-activation occurs. The E2 protein binds directly to the death effector domains (DED) of caspase 8 through non-DFD interaction. This interaction is independent of FADD, but it can cooperate with FADD homotypic binding to caspase 8 to induce its oligomerization; hence cell death, while it is antagonized by competitive binding of MC159 FLICE inhibitory protein. The amino terminal domain of E2 contains a 27 amino-acid alpha-helix, which is necessary and sufficient to induce caspase oligomerization and cell death. Our results provide evidence for adaptor-independent oligomerization of caspase 8, mediated by non-DFD direct interactions with the HPV18 E2 protein, thus deciphering a new pathway for caspase 8 activation. PMID- 18421301 TI - Role of non-canonical Beclin 1-independent autophagy in cell death induced by resveratrol in human breast cancer cells. AB - Resveratrol, a polyphenol found in grapes and other fruit and vegetables, is a powerful chemopreventive and chemotherapeutic molecule potentially of interest for the treatment of breast cancer. The human breast cancer cell line MCF-7, which is devoid of caspase-3 activity, is refractory to apoptotic cell death after incubation with resveratrol. Here we show that resveratrol arrests cell proliferation, triggers death and decreases the number of colonies of cells that are sensitive to caspase-3-dependent apoptosis (MCF-7 casp-3) and also those that are unresponsive to it (MCF-7vc). We demonstrate that resveratrol (i) acts via multiple pathways to trigger cell death, (ii) induces caspase-dependent and caspase-independent cell death in MCF-7 casp-3 cells, (iii) induces only caspase independent cell death in MCF-7vc cells and (iv) stimulates macroautophagy. Using BECN1 and hVPS34 (human vacuolar protein sorting 34) small interfering RNAs, we demonstrate that resveratrol activates Beclin 1-independent autophagy in both cell lines, whereas cell death via this uncommon form of autophagy occurs only in MCF-7vc cells. We also show that this variant form of autophagic cell death is blocked by the expression of caspase-3, but not by its enzymatic activity. In conclusion, this study reveals that non-canonical autophagy induced by resveratrol can act as a caspase-independent cell death mechanism in breast cancer cells. PMID- 18421302 TI - DNA damage-inducing agents elicit gamma-secretase activation mediated by oxidative stress. AB - According to the amyloid cascade hypothesis, Alzheimer's disease is the consequence of neuronal cell death induced by beta-amyloid (Abeta), which accumulates by abnormal clearance or production. On the other hand, recent studies have shown cell death-induced alteration in amyloid precursor protein (APP) processing, suggesting potential mutual interactions between APP processing and cell death. We have shown previously that the cell death caused by DNA damage inducing agents (DDIAs) facilitated gamma-secretase activity and Abeta generation in a Bax/Bcl-2-dependent, but caspase-independent manner. Here, we attempted to elucidate the downstream mechanism that modulates gamma-secretase activity in DDIA-treated cells. N-acetyl cysteine, a potent antioxidant, attenuated DDIA induced enhancement of gamma-secretase activity but failed to rescue cell death. Overexpression of heat shock protein 70, which blocks cytochrome c release from mitochondria, also reduced gamma-secretase activity. Moreover, glutathione depletion significantly facilitated gamma-secretase activity and Abeta generation by enhancing the formation of higher molecular weight gamma-secretase complex before signs of cell death developed. Finally, Abeta treatment, a known inducer of oxidative stress, also increased gamma-secretase activity. Taken together, these results indicate that DDIA-induced gamma-secretase activation is dependent on augmented oxidative stress, and that Abeta and gamma-secretase may activate each other. On the basis of these results, we propose a feed-back loop between oxidative stress and Abeta generation mediated by gamma-secretase activation. PMID- 18421303 TI - Brn-3a/POU4F1 interacts with and differentially affects p73-mediated transcription. AB - The Brn-3a/POU4F1 POU transcription factor is critical for the survival and differentiation of specific sensory neurons during development or upon injury; by regulating expression of target genes, either directly or indirectly upon interaction with other proteins. In this study, we demonstrated the physical interaction of Brn-3a with different p73 isoforms and showed co-localization in sensory neurons arising from the neural crest. The biological effects of p73/ Brn 3a interaction depend on the particular p73 isoform, because co-expression of Brn 3a with TAp73 enhanced cell cycle arrest, whereas Brn-3a and DeltaNp73 cooperated to increase protection from apoptosis. Brn-3a antagonized TAp73 transactivation of pro-apoptotic Bax, but co-operated to increase transcription of the cell cycle regulator p21 CIP1/Waf1. The region 425-494 amino acids within the TAp73 C terminus were critical for Brn-3a to repress Bax transactivation, but not for cooperation on the p21 CIP1/Waf1 promoter. Our results suggest that co-factors binding to the p73 C terminus facilitate maximal activation on the Bax but not p21 CIP1/Waf1 promoter and that Brn-3a modulates this interaction. Thus, the physical interaction of Brn-3a with specific p73 isoforms will be critical for determining cell fate during neuronal development or in injured neurons expressing both factors. PMID- 18421304 TI - Molecular pathogenesis of T-cell leukaemia and lymphoma. AB - T-cell acute lymphoblastic leukaemia (T-ALL) is induced by the transformation of T-cell progenitors and mainly occurs in children and adolescents. Although treatment outcome in patients with T-ALL has improved in recent years, patients with relapsed disease continue to have a poor prognosis. It is therefore important to understand the molecular pathways that control both the induction of transformation and the treatment of relapsed disease. In this Review, we focus on the molecular mechanisms responsible for disease induction and maintenance. We also compare the physiological progression of T-cell differentiation with T-cell transformation, highlighting the close relationship between these two processes. Finally, we discuss potential new therapies that target oncogenic pathways in T ALL. PMID- 18421306 TI - Targeting sodium/iodide symporter gene expression for estrogen-regulated imaging and therapy in breast cancer. AB - Expression of the sodium iodide symporter (hNIS) has been detected in breast cancer tissue, but frequently, not at the levels necessary to mediate (131)I accumulation. Transducing the hNIS gene into breast cancer cells with adenovirus could be a tractable strategy to render breast cancer susceptible to radioiodide therapy. We constructed the replication-incompetent virus, AdSERE, in which an estrogen-responsive promoter directs the expression of hNIS. In vitro, we demonstrate that AdSERE mediates hNIS expression and iodide uptake in ER+ breast cancer cells. In vivo, we show that AdSERE-infected ER+ tumors can be imaged due to tracer accumulation; in addition, AdSERE in combination with therapeutic doses of (131)I suppresses tumor growth. PMID- 18421307 TI - Effects of lentivirus-mediated HIF-1alpha knockdown on hypoxia-related cisplatin resistance and their dependence on p53 status in fibrosarcoma cells. AB - Therapy targeting hypoxia-inducible factor-1 (HIF-1) to reverse the hypoxia related drug resistance has received much interest. Despite a close interaction between HIF-1 and p53 and that p53 mutation is seen in >50% of tumors, whether HIF-1 silencing by targeted therapy depends on tumor p53 status remains unknown. Two isogenic fibrosarcoma cells HT1080 (wild-type p53) and HT1080-6TG (mutant p53) were transduced with HIF-1alpha-specific RNAi lentiviral vectors and selected with blasticidin. Real-time PCR and western blot analysis of HIF-1alpha mRNA and protein respectively validated the silencing effects. Cells were first preconditioned under hypoxia (0.5% O(2)) for 4 h and then co-treated with cisplatin for another 24 h. MTT was used for assessment of chemosensitivity to cisplatin. Moreover, annexin V and propidium iodide staining was detected on flow cytometry for analysis of cisplatin-induced apoptosis. Furthermore, changes of some Bcl-2 family members were detected on western blotting. Exposure to hypoxia significantly increased resistance to cisplatin than exposure to normoxia. HIF 1alpha knockdown could reverse hypoxia-related resistance to cisplatin and apoptotic resistance only in HT1080 cells, but had little effect on HT1080-6TG cells. With HIF-1alpha knockdown, Bid expression was higher in HT1080 than in HT1080-6TG under hypoxia. In summary, HIF-1 targeted therapy to reverse hypoxia related cisplatin resistance depends on normal p53 status. Changes of Bid expression levels under hypoxia might contribute in part to the differential response to HIF-1alpha silencing in cells with different p53 status. PMID- 18421305 TI - Immunobiology of the TAM receptors. AB - Recent studies have revealed that the TAM receptor protein tyrosine kinases- TYRO3, AXL and MER--have pivotal roles in innate immunity. They inhibit inflammation in dendritic cells and macrophages, promote the phagocytosis of apoptotic cells and membranous organelles, and stimulate the maturation of natural killer cells. Each of these phenomena may depend on a cooperative interaction between TAM receptor and cytokine receptor signalling systems. Although its importance was previously unrecognized, TAM signalling promises to have an increasingly prominent role in studies of innate immune regulation. PMID- 18421308 TI - A chimeric fusion of the hASH1 and EZH2 promoters mediates high and specific reporter and suicide gene expression and cytotoxicity in small cell lung cancer cells. AB - Transcriptionally targeted gene therapy is a promising experimental modality for treatment of systemic malignancies such as small cell lung cancer (SCLC). We have identified the human achaete-scute homolog 1 (hASH1) and enhancer of zeste homolog 2 (EZH2) genes as highly upregulated in SCLC compared to a panel of representative normal tissues. Here, we evaluate the use of regulatory regions from the hASH1- and EZH2-promoter regions alone and in combination for suicide gene therapy of SCLC. Two hASH1-promoter regions comprising 0.3 and 0.7 kb immediately upstream of (and including) the transcription start site were tested. Both constructs induced reporter gene activity (up to sevenfold SV40-promoter activity) in all tested classic (hASH1 positive) SCLC and in two hASH1-negative SCLC cell lines, whereas gene activity was low or absent (<4% of SV40 activity) in one hASH1-negative SCLC and in all control cell lines tested. To evaluate its therapeutic potential, the 0.7 kb hASH1 proximal-promoter region was evaluated for cytotoxicity in a suicide gene assay. The construct induced SCLC cytotoxicity at levels equivalent to those observed with the SV40 promoter, while control cells remained unaffected by the treatment. Analogously, a 1.1 kb EZH2-promoter region was evaluated by reporter and suicide gene assays. The EZH2 promoter potently induced reporter gene activity in SCLC (up to 25-fold of SV40 activity) while moderate reporter activity (up to 12% of SV40 activity), was detected in the control cells. However, in the suicide gene assay both control and SCLC cells demonstrated sensitivity indicating lack of promoter specificity. Finally, we fused the 0.7 kb hASH1 promoter to the EZH2 promoter generating a chimeric hASH1EZH2 regulatory construct. The chimeric promoter demonstrated increased activity in SCLC cells compared to the hASH1 promoter alone while retaining specificity in control cells. The hASH1EZH2 promoter thus constitutes a promising transcriptional regulator for SCLC gene therapy. PMID- 18421310 TI - Antiangiogenic systemic gene therapy combined with doxorubicin administration induced caspase 8 and 9-mediated apoptosis in endothelial cells and an anti metastasis effect. AB - Ad-PPE-Fas-c is an adenovector that expresses Fas-c under the control of the modified pre-proendothelin-1 (PPE-1) promoter. Fas-c is a chimeric death receptor containing the extracellular portion of tumour necrosis factor 1 receptor (TNFR1) and the transmembrane and intracellular portion of Fas. We recently demonstrated that Ad-PPE-Fas-c induced Fas-receptor-mediated endothelial cell apoptosis. Previously, doxorubicin was shown to enhance Fas-receptor clustering and the induction of its cascade. Therefore, the goal of this work was to test whether doxorubicin augments the capacity of Ad-PPE-Fas-c to induce endothelial cell apoptosis and to elucidate whether either the death-receptor-mediated apoptotic cascade or the mitochondria-associated apoptotic cascade is involved in the combined treatment effect. We found that a combined treatment of Ad-PPE-Fas-c and doxorubicin synergistically induced a reduction in endothelial cell viability and apoptosis. z-IETD-FMK, a caspase-8 inhibitor, and z-LEHD-FMK, a caspase-9 inhibitor, significantly decreased apoptosis induced by the combined treatment. Systemically administered combined therapy significantly reduced the lung metastases burden (70%) in mice as compared to each treatment alone. Thus, a combined treatment of Ad-PPE-Fas-c gene therapy and chemotherapy may be effective in the treatment of metastatic diseases and both the Fas cascade and the mitochondria-associated cascade are essential for this effect. PMID- 18421309 TI - Primary neural stem/progenitor cells expressing endostatin or cytochrome P450 for gene therapy of glioblastoma. AB - Despite recent technical improvements in surgical excision techniques and adjuvant radio- and chemotherapy, the clinical outcome of patients with grade IV astrocytoma (glioblastoma) remains very poor, with a median survival of less than 12 months. A promising approach to therapy employs gene-engineered neural stem/progenitor cells (NSCs) as a cellular therapeutic delivery system, to track glioblastoma cells and deliver anticancer molecules. However, most results on their tumor tropism have been derived by immortalized NSCs. We now report that primary murine gene-engineered NSCs displayed in vivo tropism for glioblastoma cells. Ten days after injection into the brain, many NSCs continued to express the transgene and the NSC marker, nestin. NSCs transduced with a retroviral vector co-expressing a secretable form of human endostatin and eGFP (NSC/endo eGFP) released potentially antiangiogenetic concentrations of endostatin into the culture medium. Conditioned medium of NSC/endo-eGFP cells inhibited the growth of mouse pulmonary microvascular endothelial cells (PMVECs). A good correlation between endostatin levels and PMVEC growth inhibition was observed. In NSCs co expressing cytochrome P450 2B6 (CYP2B6) and eGFP (NSC/CYP2B6-eGFP), the forced expression of CYP2B6 resulted in intracellular activation of CPA and subsequent cell death. In the presence of NSC/CYP2B6-eGFP, we observed CPA cytotoxicity to DsRed-expressing U87Mg glioma cells. In vivo treatment of intracranial GL-261 glioblastoma with NSC/endo-eGFP caused a 65% reduction in tumor size, compared to untreated control mice or mice treated with NSC/eGFP cells. Our data suggest that primary NSCs transduced with retroviral vectors expressing endostatin and/or CYP2B6 have a potential role in glioblastoma therapy. PMID- 18421311 TI - Vaccination of fiber-modified adenovirus-transfected dendritic cells to express HER-2/neu stimulates efficient HER-2/neu-specific humoral and CTL responses and reduces breast carcinogenesis in transgenic mice. AB - HER-2/neu transgene-modified dendritic cell (DC)-based vaccines are potent at eliciting HER-2/neu-specific antitumor immunity. In this study, we constructed a recombinant adenovirus (RGD)AdVneu with fiber gene modified by RGD insertion into the viral knob's H1 loop. We transfected DCs with (RGD)AdVneu, and assessed/compared HER-2/neu-specific humoral and cytotoxic T lymphocyte (CTL) responses and antitumor immunity derived from the original AdVneu-transfected DCs (DCneu1) and (RGD)AdVneu-transfected DCs (DCneu2). We demonstrated that DCneu2 displayed increased HER-2/neu expression by 8.3-fold compared to DCneu1. We also demonstrated that DCneu2 vaccination induced stronger HER-2/neu-specific humoral and CTL immune responses than DCneu1 vaccination. DCneu2 vaccination protected all the mice from HER-2/neu-expressing Tg1-1 tumor cell challenge in wild-type FVB/NJ mice, compared to a partial protection in DCneu1-immunized mice. In addition, DCneu2 vaccination also significantly delayed tumor growth than DCneu1 immunization (P<0.05) in Tg FVBneuN mice. Three immunizations of DCneu2 starting at the mouse age of 2 months also significantly delayed breast cancer development in Tg mice compared to DCneu2 vaccine (P<0.05). Importantly, DCneu2 vaccine reduced breast carcinogenesis by 9% in Tg mice with self HER-2/neu tolerance. Therefore, vaccination of fiber-modified adenovirus-transfected DCs to enhance expression of tumor antigens such as HER-2/neu is likely representative of a new direction in DC-based vaccine of breast cancer. PMID- 18421312 TI - A universal classification of eukaryotic transposable elements implemented in Repbase. PMID- 18421317 TI - A ghost of battles past. PMID- 18421318 TI - All eyes north. PMID- 18421319 TI - Broken promises. PMID- 18421322 TI - Novel analysis identifies highly biodiverse hotspots. PMID- 18421324 TI - Tibetan wildlife is getting used to the railway. PMID- 18421325 TI - Malaria: efforts starting to show widespread results. PMID- 18421326 TI - Malaria programmes need informed advocacy. PMID- 18421327 TI - NIH: researchers lose out to war, not to each other. PMID- 18421328 TI - NIH: grants revamp needs grounding in evidence. PMID- 18421329 TI - NIH funds support more than a researcher's own lab. PMID- 18421330 TI - Vaccine trial provided valuable information. PMID- 18421336 TI - Triumph of the medieval mind. PMID- 18421337 TI - Human genetics: Dr Watson's base pairs. PMID- 18421339 TI - Quantum physics: debut of the quarter electron. PMID- 18421338 TI - Planetary science: message from Mercury. PMID- 18421340 TI - Biophysics: the sweetest candy for the virus. PMID- 18421342 TI - Systems biology: genome rewired. PMID- 18421343 TI - Ecology: destabilized fish stocks. PMID- 18421344 TI - Astrophysics: blown away by cosmic rays. PMID- 18421345 TI - Observation of a quarter of an electron charge at the nu = 5/2 quantum Hall state. AB - The fractional quantum Hall effect, where plateaus in the Hall resistance at values of h/nue2 coexist with zeros in the longitudinal resistance, results from electron correlations in two dimensions under a strong magnetic field. (Here h is Planck's constant, nu the filling factor and e the electron charge.) Current flows along the sample edges and is carried by charged excitations (quasiparticles) whose charge is a fraction of the electron charge. Although earlier research concentrated on odd denominator fractional values of nu, the observation of the even denominator nu = 5/2 state sparked much interest. This state is conjectured to be characterized by quasiparticles of charge e/4, whose statistics are 'non-abelian'-in other words, interchanging two quasiparticles may modify the state of the system into a different one, rather than just adding a phase as is the case for fermions or bosons. As such, these quasiparticles may be useful for the construction of a topological quantum computer. Here we report data on shot noise generated by partitioning edge currents in the nu = 5/2 state, consistent with the charge of the quasiparticle being e/4, and inconsistent with other possible values, such as e/2 and e. Although this finding does not prove the non-abelian nature of the nu = 5/2 state, it is the first step towards a full understanding of these new fractional charges. PMID- 18421346 TI - Why fishing magnifies fluctuations in fish abundance. AB - It is now clear that fished populations can fluctuate more than unharvested stocks. However, it is not clear why. Here we distinguish among three major competing mechanisms for this phenomenon, by using the 50-year California Cooperative Oceanic Fisheries Investigations (CalCOFI) larval fish record. First, variable fishing pressure directly increases variability in exploited populations. Second, commercial fishing can decrease the average body size and age of a stock, causing the truncated population to track environmental fluctuations directly. Third, age-truncated or juvenescent populations have increasingly unstable population dynamics because of changing demographic parameters such as intrinsic growth rates. We find no evidence for the first hypothesis, limited evidence for the second and strong evidence for the third. Therefore, in California Current fisheries, increased temporal variability in the population does not arise from variable exploitation, nor does it reflect direct environmental tracking. More fundamentally, it arises from increased instability in dynamics. This finding has implications for resource management as an empirical example of how selective harvesting can alter the basic dynamics of exploited populations, and lead to unstable booms and busts that can precede systematic declines in stock levels. PMID- 18421348 TI - A massive binary black-hole system in OJ 287 and a test of general relativity. AB - Tests of Einstein's general theory of relativity have mostly been carried out in weak gravitational fields where the space-time curvature effects are first-order deviations from Newton's theory. Binary pulsars provide a means of probing the strong gravitational field around a neutron star, but strong-field effects may be best tested in systems containing black holes. Here we report such a test in a close binary system of two candidate black holes in the quasar OJ 287. This quasar shows quasi-periodic optical outbursts at 12-year intervals, with two outburst peaks per interval. The latest outburst occurred in September 2007, within a day of the time predicted by the binary black-hole model and general relativity. The observations confirm the binary nature of the system and also provide evidence for the loss of orbital energy in agreement (within 10 per cent) with the emission of gravitational waves from the system. In the absence of gravitational wave emission the outburst would have happened 20 days later. PMID- 18421347 TI - Evolvability and hierarchy in rewired bacterial gene networks. AB - Sequencing DNA from several organisms has revealed that duplication and drift of existing genes have primarily moulded the contents of a given genome. Though the effect of knocking out or overexpressing a particular gene has been studied in many organisms, no study has systematically explored the effect of adding new links in a biological network. To explore network evolvability, we constructed 598 recombinations of promoters (including regulatory regions) with different transcription or sigma-factor genes in Escherichia coli, added over a wild-type genetic background. Here we show that approximately 95% of new networks are tolerated by the bacteria, that very few alter growth, and that expression level correlates with factor position in the wild-type network hierarchy. Most importantly, we find that certain networks consistently survive over the wild type under various selection pressures. Therefore new links in the network are rarely a barrier for evolution and can even confer a fitness advantage. PMID- 18421349 TI - Thermalization and its mechanism for generic isolated quantum systems. AB - An understanding of the temporal evolution of isolated many-body quantum systems has long been elusive. Recently, meaningful experimental studies of the problem have become possible, stimulating theoretical interest. In generic isolated systems, non-equilibrium dynamics is expected to result in thermalization: a relaxation to states in which the values of macroscopic quantities are stationary, universal with respect to widely differing initial conditions, and predictable using statistical mechanics. However, it is not obvious what feature of many-body quantum mechanics makes quantum thermalization possible in a sense analogous to that in which dynamical chaos makes classical thermalization possible. For example, dynamical chaos itself cannot occur in an isolated quantum system, in which the time evolution is linear and the spectrum is discrete. Some recent studies even suggest that statistical mechanics may give incorrect predictions for the outcomes of relaxation in such systems. Here we demonstrate that a generic isolated quantum many-body system does relax to a state well described by the standard statistical-mechanical prescription. Moreover, we show that time evolution itself plays a merely auxiliary role in relaxation, and that thermalization instead happens at the level of individual eigenstates, as first proposed by Deutsch and Srednicki. A striking consequence of this eigenstate thermalization scenario, confirmed for our system, is that knowledge of a single many-body eigenstate is sufficient to compute thermal averages-any eigenstate in the microcanonical energy window will do, because they all give the same result. PMID- 18421350 TI - Electrical effects of spin density wave quantization and magnetic domain walls in chromium. AB - The role of magnetic domains (and the walls between domains) in determining the electrical properties of ferromagnetic materials has been investigated in great detail for many years, not least because control over domains offers a means of manipulating electron spin to control charge transport in 'spintronic' devices. In contrast, much less attention has been paid to the effects of domains and domain walls on the electrical properties of antiferromagnets: antiferromagnetic domains show no net external magnetic moment, and so are difficult to manipulate or probe. Here we describe electrical measurements on chromium--a simple metal and quintessential spin density wave antiferromagnet--that show behaviour directly related to spin density wave formation and the presence of antiferromagnetic domains. Two types of thermal hysteresis are seen in both longitudinal and Hall resistivity: the first can be explained by the quantization of spin density waves due to the finite film thickness (confirmed by X-ray diffraction measurements) and the second by domain-wall scattering of electrons. We also observe the striking influence of the electrical lead configuration (a mesoscopic effect) on the resistivity of macroscopic samples in the spin density wave state. Our results are potentially of practical importance, in that they reveal tunable electrical effects of film thickness and domain walls that are as large as the highest seen for ferromagnets. PMID- 18421351 TI - Changing boreal methane sources and constant biomass burning during the last termination. AB - Past atmospheric methane concentrations show strong fluctuations in parallel to rapid glacial climate changes in the Northern Hemisphere superimposed on a glacial-interglacial doubling of methane concentrations. The processes driving the observed fluctuations remain uncertain but can be constrained using methane isotopic information from ice cores. Here we present an ice core record of carbon isotopic ratios in methane over the entire last glacial-interglacial transition. Our data show that the carbon in atmospheric methane was isotopically much heavier in cold climate periods. With the help of a box model constrained by the present data and previously published results, we are able to estimate the magnitude of past individual methane emission sources and the atmospheric lifetime of methane. We find that methane emissions due to biomass burning were about 45 Tg methane per year, and that these remained roughly constant throughout the glacial termination. The atmospheric lifetime of methane is reduced during cold climate periods. We also show that boreal wetlands are an important source of methane during warm events, but their methane emissions are essentially shut down during cold climate conditions. PMID- 18421352 TI - The complete genome of an individual by massively parallel DNA sequencing. AB - The association of genetic variation with disease and drug response, and improvements in nucleic acid technologies, have given great optimism for the impact of 'genomic medicine'. However, the formidable size of the diploid human genome, approximately 6 gigabases, has prevented the routine application of sequencing methods to deciphering complete individual human genomes. To realize the full potential of genomics for human health, this limitation must be overcome. Here we report the DNA sequence of a diploid genome of a single individual, James D. Watson, sequenced to 7.4-fold redundancy in two months using massively parallel sequencing in picolitre-size reaction vessels. This sequence was completed in two months at approximately one-hundredth of the cost of traditional capillary electrophoresis methods. Comparison of the sequence to the reference genome led to the identification of 3.3 million single nucleotide polymorphisms, of which 10,654 cause amino-acid substitution within the coding sequence. In addition, we accurately identified small-scale (2-40,000 base pair (bp)) insertion and deletion polymorphism as well as copy number variation resulting in the large-scale gain and loss of chromosomal segments ranging from 26,000 to 1.5 million base pairs. Overall, these results agree well with recent results of sequencing of a single individual by traditional methods. However, in addition to being faster and significantly less expensive, this sequencing technology avoids the arbitrary loss of genomic sequences inherent in random shotgun sequencing by bacterial cloning because it amplifies DNA in a cell-free system. As a result, we further demonstrate the acquisition of novel human sequence, including novel genes not previously identified by traditional genomic sequencing. This is the first genome sequenced by next-generation technologies. Therefore it is a pilot for the future challenges of 'personalized genome sequencing'. PMID- 18421356 TI - Nanotechnology: could it be a small world after all? PMID- 18421364 TI - Analysis of chimpanzee history based on genome sequence alignments. AB - Population geneticists often study small numbers of carefully chosen loci, but it has become possible to obtain orders of magnitude for more data from overlaps of genome sequences. Here, we generate tens of millions of base pairs of multiple sequence alignments from combinations of three western chimpanzees, three central chimpanzees, an eastern chimpanzee, a bonobo, a human, an orangutan, and a macaque. Analysis provides a more precise understanding of demographic history than was previously available. We show that bonobos and common chimpanzees were separated approximately 1,290,000 years ago, western and other common chimpanzees approximately 510,000 years ago, and eastern and central chimpanzees at least 50,000 years ago. We infer that the central chimpanzee population size increased by at least a factor of 4 since its separation from western chimpanzees, while the western chimpanzee effective population size decreased. Surprisingly, in about one percent of the genome, the genetic relationships between humans, chimpanzees, and bonobos appear to be different from the species relationships. We used PCR-based resequencing to confirm 11 regions where chimpanzees and bonobos are not most closely related. Study of such loci should provide information about the period of time 5-7 million years ago when the ancestors of humans separated from those of the chimpanzees. PMID- 18421365 TI - Letter to the Editor. PMID- 18421366 TI - Response to the letter to the Editor. PMID- 18421367 TI - CME for child psychiatrists: recommendations for learners, planners and presenters. AB - INTRODUCTION: Medical school and residency are only the beginning of a child psychiatrist's education. For the rest of her/his career, a child psychiatrist will need to learn on an ongoing basis. There will always be new understandings, new treatments, new issues to master. Child psychiatrists will always need to further their knowledge, develop new skills, and improve existing skills. For these reasons at very least, all child psychiatrists will need to participate in Continuing Medical Education (CME) activities. Many child psychiatrists will also be involved in the design and delivery of these CME activities. In both cases, understanding more about the effectiveness of CME will be important to the decisions they make. METHOD: This article itself is not a systematic review of the literature, but it will highlight some of the important findings from existing systematic reviews of the CME literature. Based on these findings, the article will make recommendations for both child psychiatrists as learners and child psychiatrists as CME presenters. RESULTS: As learners, child psychiatrists need to be able to select CME activities that are most likely to lead to improvements in their practices. As planners and presenters, child psychiatrists need to design and deliver CME activities that are most likely to improve the practices of their target audiences. However, not all child psychiatrists have the time to review the CME literature in addition to reviewing the other bodies of literature relevant to their practices. CONCLUSION: Thus, the purpose of this article is to provide an overview of the key findings in the CME literature, focusing on the effectiveness of CME. PMID- 18421368 TI - Empirical testing of an algorithm for defining somatization in children. AB - INTRODUCTION: A previous article proposed an algorithm for defining somatization in children by classifying them into three categories: well, medically ill, and somatizer; the authors suggested further empirical validation of the algorithm (Postilnik et al., 2006). We use the Child Behavior Checklist (CBCL) to provide this empirical validation. METHOD: Parents of children seen in pediatric clinics completed the CBCL (n=126). The physicians of these children completed specially designed questionnaires. The sample comprised of 62 boys and 64 girls (age range 2 to 15 years). Classification categories included: well (n=53), medically ill (n=55), and somatizer (n=18). Analysis of variance (ANOVA) was used for statistical comparisons. Discriminant function analysis was conducted with the CBCL subscales. RESULTS: There were significant differences between the classification categories for the somatic complaints (p=<0.001), social problems (p=0.004), thought problems (p=0.01), attention problems (0.006), and internalizing (p=0.003) subscales and also total (p=0.001), and total-t (p=0.001) scales of the CBCL. Discriminant function analysis showed that 78% of somatizers and 66% of well were accurately classified, while only 35% of medically ill were accurately classified. CONCLUSION: The somatization classification algorithm proposed by Postilnik et al. (2006) shows promise for classification of children and adolescents with somatic symptoms. PMID- 18421369 TI - Certain eating disorders may be a neuropsychiatric manifestation of PANDAS: case report. PMID- 18421370 TI - Commentary on clinical case rounds in child and adolescent psychiatry. PMID- 18421371 TI - Predicting co-complexed protein pairs from heterogeneous data. AB - Proteins do not carry out their functions alone. Instead, they often act by participating in macromolecular complexes and play different functional roles depending on the other members of the complex. It is therefore interesting to identify co-complex relationships. Although protein complexes can be identified in a high-throughput manner by experimental technologies such as affinity purification coupled with mass spectrometry (APMS), these large-scale datasets often suffer from high false positive and false negative rates. Here, we present a computational method that predicts co-complexed protein pair (CCPP) relationships using kernel methods from heterogeneous data sources. We show that a diffusion kernel based on random walks on the full network topology yields good performance in predicting CCPPs from protein interaction networks. In the setting of direct ranking, a diffusion kernel performs much better than the mutual clustering coefficient. In the setting of SVM classifiers, a diffusion kernel performs much better than a linear kernel. We also show that combination of complementary information improves the performance of our CCPP recognizer. A summation of three diffusion kernels based on two-hybrid, APMS, and genetic interaction networks and three sequence kernels achieves better performance than the sequence kernels or diffusion kernels alone. Inclusion of additional features achieves a still better ROC(50) of 0.937. Assuming a negative-to-positive ratio of 600ratio1, the final classifier achieves 89.3% coverage at an estimated false discovery rate of 10%. Finally, we applied our prediction method to two recently described APMS datasets. We find that our predicted positives are highly enriched with CCPPs that are identified by both datasets, suggesting that our method successfully identifies true CCPPs. An SVM classifier trained from heterogeneous data sources provides accurate predictions of CCPPs in yeast. This computational method thereby provides an inexpensive method for identifying protein complexes that extends and complements high-throughput experimental data. PMID- 18421372 TI - Investigations of oligonucleotide usage variance within and between prokaryotes. AB - Oligonucleotide usage in archaeal and bacterial genomes can be linked to a number of properties, including codon usage (trinucleotides), DNA base-stacking energy (dinucleotides), and DNA structural conformation (di- to tetranucleotides). We wanted to assess the statistical information potential of different DNA 'word sizes' and explore how oligonucleotide frequencies differ in coding and non coding regions. In addition, we used oligonucleotide frequencies to investigate DNA composition and how DNA sequence patterns change within and between prokaryotic organisms. Among the results found was that prokaryotic chromosomes can be described by hexanucleotide frequencies, suggesting that prokaryotic DNA is predominantly short range correlated, i.e., information in prokaryotic genomes is encoded in short oligonucleotides. Oligonucleotide usage varied more within AT rich and host-associated genomes than in GC-rich and free-living genomes, and this variation was mainly located in non-coding regions. Bias (selectional pressure) in tetranucleotide usage correlated with GC content, and coding regions were more biased than non-coding regions. Non-coding regions were also found to be approximately 5.5% more AT-rich than coding regions, on average, in the 402 chromosomes examined. Pronounced DNA compositional differences were found both within and between AT-rich and GC-rich genomes. GC-rich genomes were more similar and biased in terms of tetranucleotide usage in non-coding regions than AT-rich genomes. The differences found between AT-rich and GC-rich genomes may possibly be attributed to lifestyle, since tetranucleotide usage within host-associated bacteria was, on average, more dissimilar and less biased than free-living archaea and bacteria. PMID- 18421373 TI - Synaptic learning rules and sparse coding in a model sensory system. AB - Neural circuits exploit numerous strategies for encoding information. Although the functional significance of individual coding mechanisms has been investigated, ways in which multiple mechanisms interact and integrate are not well understood. The locust olfactory system, in which dense, transiently synchronized spike trains across ensembles of antenna lobe (AL) neurons are transformed into a sparse representation in the mushroom body (MB; a region associated with memory), provides a well-studied preparation for investigating the interaction of multiple coding mechanisms. Recordings made in vivo from the insect MB demonstrated highly specific responses to odors in Kenyon cells (KCs). Typically, only a few KCs from the recorded population of neurons responded reliably when a specific odor was presented. Different odors induced responses in different KCs. Here, we explored with a biologically plausible model the possibility that a form of plasticity may control and tune synaptic weights of inputs to the mushroom body to ensure the specificity of KCs' responses to familiar or meaningful odors. We found that plasticity at the synapses between the AL and the MB efficiently regulated the delicate tuning necessary to selectively filter the intense AL oscillatory output and condense it to a sparse representation in the MB. Activity-dependent plasticity drove the observed specificity, reliability, and expected persistence of odor representations, suggesting a role for plasticity in information processing and making a testable prediction about synaptic plasticity at AL-MB synapses. PMID- 18421374 TI - Functional maps of protein complexes from quantitative genetic interaction data. AB - Recently, a number of advanced screening technologies have allowed for the comprehensive quantification of aggravating and alleviating genetic interactions among gene pairs. In parallel, TAP-MS studies (tandem affinity purification followed by mass spectroscopy) have been successful at identifying physical protein interactions that can indicate proteins participating in the same molecular complex. Here, we propose a method for the joint learning of protein complexes and their functional relationships by integration of quantitative genetic interactions and TAP-MS data. Using 3 independent benchmark datasets, we demonstrate that this method is >50% more accurate at identifying functionally related protein pairs than previous approaches. Application to genes involved in yeast chromosome organization identifies a functional map of 91 multimeric complexes, a number of which are novel or have been substantially expanded by addition of new subunits. Interestingly, we find that complexes that are enriched for aggravating genetic interactions (i.e., synthetic lethality) are more likely to contain essential genes, linking each of these interactions to an underlying mechanism. These results demonstrate the importance of both large-scale genetic and physical interaction data in mapping pathway architecture and function. PMID- 18421375 TI - Performance and scalability of discriminative metrics for comparative gene identification in 12 Drosophila genomes. AB - Comparative genomics of multiple related species is a powerful methodology for the discovery of functional genomic elements, and its power should increase with the number of species compared. Here, we use 12 Drosophila genomes to study the power of comparative genomics metrics to distinguish between protein-coding and non-coding regions. First, we study the relative power of different comparative metrics and their relationship to single-species metrics. We find that even relatively simple multi-species metrics robustly outperform advanced single species metrics, especially for shorter exons (< or =240 nt), which are common in animal genomes. Moreover, the two capture largely independent features of protein coding genes, with different sensitivity/specificity trade-offs, such that their combinations lead to even greater discriminatory power. In addition, we study how discovery power scales with the number and phylogenetic distance of the genomes compared. We find that species at a broad range of distances are comparably effective informants for pairwise comparative gene identification, but that these are surpassed by multi-species comparisons at similar evolutionary divergence. In particular, while pairwise discovery power plateaued at larger distances and never outperformed the most advanced single-species metrics, multi-species comparisons continued to benefit even from the most distant species with no apparent saturation. Last, we find that genes in functional categories typically considered fast-evolving can nonetheless be recovered at very high rates using comparative methods. Our results have implications for comparative genomics analyses in any species, including the human. PMID- 18421376 TI - An unbiased systems genetics approach to mapping genetic loci modulating susceptibility to severe streptococcal sepsis. AB - Striking individual differences in severity of group A streptococcal (GAS) sepsis have been noted, even among patients infected with the same bacterial strain. We had provided evidence that HLA class II allelic variation contributes significantly to differences in systemic disease severity by modulating host responses to streptococcal superantigens. Inasmuch as the bacteria produce additional virulence factors that participate in the pathogenesis of this complex disease, we sought to identify additional gene networks modulating GAS sepsis. Accordingly, we applied a systems genetics approach using a panel of advanced recombinant inbred mice. By analyzing disease phenotypes in the context of mice genotypes we identified a highly significant quantitative trait locus (QTL) on Chromosome 2 between 22 and 34 Mb that strongly predicts disease severity, accounting for 25%-30% of variance. This QTL harbors several polymorphic genes known to regulate immune responses to bacterial infections. We evaluated candidate genes within this QTL using multiple parameters that included linkage, gene ontology, variation in gene expression, cocitation networks, and biological relevance, and identified interleukin1 alpha and prostaglandin E synthases pathways as key networks involved in modulating GAS sepsis severity. The association of GAS sepsis with multiple pathways underscores the complexity of traits modulating GAS sepsis and provides a powerful approach for analyzing interactive traits affecting outcomes of other infectious diseases. PMID- 18421377 TI - Chapare virus, a newly discovered arenavirus isolated from a fatal hemorrhagic fever case in Bolivia. AB - A small focus of hemorrhagic fever (HF) cases occurred near Cochabamba, Bolivia, in December 2003 and January 2004. Specimens were available from only one fatal case, which had a clinical course that included fever, headache, arthralgia, myalgia, and vomiting with subsequent deterioration and multiple hemorrhagic signs. A non-cytopathic virus was isolated from two of the patient serum samples, and identified as an arenavirus by IFA staining with a rabbit polyvalent antiserum raised against South American arenaviruses known to be associated with HF (Guanarito, Machupo, and Sabia). RT-PCR analysis and subsequent analysis of the complete virus S and L RNA segment sequences identified the virus as a member of the New World Clade B arenaviruses, which includes all the pathogenic South American arenaviruses. The virus was shown to be most closely related to Sabia virus, but with 26% and 30% nucleotide difference in the S and L segments, and 26%, 28%, 15% and 22% amino acid differences for the L, Z, N, and GP proteins, respectively, indicating the virus represents a newly discovered arenavirus, for which we propose the name Chapare virus. In conclusion, two different arenaviruses, Machupo and Chapare, can be associated with severe HF cases in Bolivia. PMID- 18421378 TI - Identification of a bacterial-like HslVU protease in the mitochondria of Trypanosoma brucei and its role in mitochondrial DNA replication. AB - ATP-dependent protease complexes are present in all living organisms, including the 26S proteasome in eukaryotes, Archaea, and Actinomycetales, and the HslVU protease in eubacteria. The structure of HslVU protease resembles that of the 26S proteasome, and the simultaneous presence of both proteases in one organism was deemed unlikely. However, HslVU homologs have been identified recently in some primordial eukaryotes, though their potential function remains elusive. We characterized the HslVU homolog from Trypanosoma brucei, a eukaryotic protozoan parasite and the causative agent of human sleeping sickness. TbHslVU has ATP dependent peptidase activity and, like its bacterial counterpart, has essential lysine and N-terminal threonines in the catalytic subunit. By epitope tagging, TbHslVU localizes to mitochondria and is associated with the mitochondrial genome, kinetoplast DNA (kDNA). RNAi of TbHslVU dramatically affects the kDNA by causing over-replication of the minicircle DNA. This leads to defects in kDNA segregation and, subsequently, to continuous network growth to an enormous size. Multiple discrete foci of nicked/gapped minicircles are formed on the periphery of kDNA disc, suggesting a failure in repairing the gaps in the minicircles for kDNA segregation. TbHslVU is a eubacterial protease identified in the mitochondria of a eukaryote. It has a novel function in regulating mitochondrial DNA replication that has never been observed in other organisms. PMID- 18421379 TI - DC-SIGN and CD150 have distinct roles in transmission of measles virus from dendritic cells to T-lymphocytes. AB - Measles virus (MV) is among the most infectious viruses that affect humans and is transmitted via the respiratory route. In macaques, MV primarily infects lymphocytes and dendritic cells (DCs). Little is known about the initial target cell for MV infection. Since DCs bridge the peripheral mucosal tissues with lymphoid tissues, we hypothesize that DCs are the initial target cells that capture MV in the respiratory tract and transport the virus to the lymphoid tissues where MV is transmitted to lymphocytes. Recently, we have demonstrated that the C-type lectin DC-SIGN interacts with MV and enhances infection of DCs in cis. Using immunofluorescence microscopy, we demonstrate that DC-SIGN+ DCs are abundantly present just below the epithelia of the respiratory tract. DC-SIGN+ DCs efficiently present MV-derived antigens to CD4+ T-lymphocytes after antigen uptake via either CD150 or DC-SIGN in vitro. However, DC-SIGN+ DCs also mediate transmission of MV to CD4+ and CD8+ T-lymphocytes. We distinguished two different transmission routes that were either dependent or independent on direct DC infection. DC-SIGN and CD150 are both involved in direct DC infection and subsequent transmission of de novo synthesized virus. However, DC-SIGN, but not CD150, mediates trans-infection of MV to T-lymphocytes independent of DC infection. Together these data suggest a prominent role for DCs during the initiation, dissemination, and clearance of MV infection. PMID- 18421380 TI - Transmission pathways of foot-and-mouth disease virus in the United Kingdom in 2007. AB - Foot-and-mouth disease (FMD) virus causes an acute vesicular disease of domesticated and wild ruminants and pigs. Identifying sources of FMD outbreaks is often confounded by incomplete epidemiological evidence and the numerous routes by which virus can spread (movements of infected animals or their products, contaminated persons, objects, and aerosols). Here, we show that the outbreaks of FMD in the United Kingdom in August 2007 were caused by a derivative of FMDV O(1) BFS 1860, a virus strain handled at two FMD laboratories located on a single site at Pirbright in Surrey. Genetic analysis of complete viral genomes generated in real-time reveals a probable chain of transmission events, predicting undisclosed infected premises, and connecting the second cluster of outbreaks in September to those in August. Complete genome sequence analysis of FMD viruses conducted in real-time have identified the initial and intermediate sources of these outbreaks and demonstrate the value of such techniques in providing information useful to contemporary disease control programmes. PMID- 18421381 TI - The synthesis of enantiomerically pure [2.2]paracyclophane derivatives. AB - [2.2]Paracyclophane is a fascinating molecule that offers great potential in a wide range of chemical disciplines. Currently, the synthesis of the majority of enantiomerically pure [2.2]paracyclophane derivatives is based on the resolution of a small number of starting materials or individual resolution procedures are developed for each new compound. The development of more general routes to these valuable compounds via the resolution of a common intermediate is discussed. Ultimately, it would be preferable to synthesise these valuable compounds without recourse to resolution and ideas for this rewarding goal are postulated. PMID- 18421382 TI - "Frustrated Lewis pairs": a concept for new reactivity and catalysis. AB - The concept of "frustrated Lewis pairs" is described and shown to result in molecular systems capable of unique reactivity as well as applications in catalysis. PMID- 18421383 TI - Enzymatic resolution and evaluation of enantiomers of cis-5'-hydroxythalidomide. AB - The straightforward synthesis of both enantiomers of cis-5'-hydroxythalidomide, a major metabolite of thalidomide, has been accomplished by enzymatic kinetic resolution of a racemic substrate catalyzed by Pseudomonas stutzeri lipase TL. cis-5'-Hydroxythalidomide shows resistance to racemization (and epimerization) at physiological pH. A tube formation assay to assess the ability to inhibit angiogenesis revealed that cis-5'-hydroxythalidomides are inactive. PMID- 18421384 TI - Photoresponsive dithienylethene-urea-based organogels with "reversed" behavior. AB - Dithienylperhydrocyclopentene-bisurea-based low molecular weight gelators are described that function as photoresponsive organogels that show a remarkable gel to-liquid transition upon irradiation. The two series of derivatives, with and without alkyl spacers between the urea hydrogen bonding groups and the photochromic unit, show different gelation behavior. Upon UV irradiation of the gels, a gel liquified at only 1.4% conversion of the photochromic unit. Transmission electron microscopy (TEM) shows that the gel fibres consist of thin ribbons. Semi-empirical (PM3) calculations indicate that the hydrogen bonding between the open-ring isomer (o) molecules is weak, and that formation of the closed-ring isomer (c) destabilises the hydrogen bonding further. The results indicate that a small amount of the closed-ring isomer will disrupt the intermolecular hydrogen-bonding, leading to disintegration of the gel fibre ribbons and hence reversible liquification. PMID- 18421385 TI - Porphyrin-bile acid conjugates: from saccharide recognition in the solution to the selective cancer cell fluorescence detection. AB - This paper describes the preparation and use of conjugates of porphyrins and bile acids as ligands to bind to tumor expressed saccharides. Bile acid-porphyrin conjugates were tested for recognition of saccharides that are typically present on malignant tumor cells. Fluorescence microscopy, in vitro PDT cell killing, and PDT of subcutaneous 4T1 mouse tumors is reported. High selectivity for saccharide cancer markers and cancer cells was observed. This in vivo and in vitro study demonstrated high potential use for these compounds in targeted photodynamic therapy. PMID- 18421386 TI - Control over binding stoichiometry and specificity in the supramolecular immobilization of cytochrome c on a molecular printboard. AB - Here, the stepwise assembly of an electroactive bionanostructure on a molecular printboard is described. The system consists of a cyclodextrin receptor monolayer (molecular printboard) on glass, a divalent linker, streptavidin (SAv), and biotinylated cytochrome c (cyt c). The divalent linker consists of a biotin moiety for binding to SAv and two adamantyl moieties for supramolecular host guest interaction at the cyclodextrin molecular printboard. The binding of biotinylated cyt c onto a SAv layer bound to preadsorbed linker appeared to be highly specific. The coverages of cyt c as assessed by UV-vis spectroscopy and scanning electrochemical microscopy (SECM) appeared to be identical indicating that all cyt c units remained active. Moreover, the coverage values corresponded well with an estimate based on steric requirements, and the binding stoichiometry was therefore found to be by two biotin moieties of cyt c per one SAv molecule. PMID- 18421387 TI - Highly effective receptors showing di- vs. monosaccharide preference. AB - Receptors and , incorporating two heterocyclic recognition units as well as oxime or hydroxymethyl-based hydrogen-bonding sites, were prepared, and their binding properties toward neutral sugars were determined. The design of these receptors was inspired by the binding motifs observed in the crystal structure of protein carbohydrate complexes. The receptors and are able to recognize both mono- and disaccharides, with a strong preference for the disaccharides. Both hydrogen bonding and interactions of the sugar CH's with the phenyl rings of the receptor contribute to the stabilisation of the receptor-sugar complexes. Molecular modeling calculations, synthesis and binding studies are described. PMID- 18421388 TI - Synthesis and structure-property relationships of donor/acceptor-functionalized bis(dehydrobenzo[18]annuleno)benzenes. AB - Seven new bis(dehydrobenzo[18]annuleno)benzenes (bis[18]DBAs) functionalized with electron-donating dibutylamino groups and/or accepting nitro groups at various positions along the peripheries of the chromophores have been prepared. The effects of varying the donor/acceptor charge transfer pathways, chromophore lengths and molecular symmetries upon the optical band gaps are studied using UV visible spectroscopy, and structure-property correlations are identified. It is found that bis[18]DBAs possessing donor-pi-donor and acceptor-pi-acceptor pathways exhibit the smallest band gaps, especially when an acceptor-pi-acceptor pathway is situated along the longest chromophore length in the molecule. The all donor species is also found to exhibit efficient fluorescence with dramatic solvatochromism. The results may have value to the rational design of future NLO/TPA device components. PMID- 18421389 TI - A modular flow reactor for performing Curtius rearrangements as a continuous flow process. AB - The use of a mesofluidic flow reactor is described for performing Curtius rearrangement reactions of carboxylic acids in the presence of diphenylphosphoryl azide and trapping of the intermediate isocyanates with various nucleophiles. PMID- 18421390 TI - Azide monoliths as convenient flow reactors for efficient Curtius rearrangement reactions. AB - The preparation and use of an azide-containing monolithic reactor is described for use in a flow chemistry device and in particular for conducting Curtius rearrangement reactions via acid chloride inputs. PMID- 18421391 TI - Proline-rich proteins--deriving a basis for residue-based selectivity in polyphenolic binding. AB - (1)H NMR titration experiments have been used to establish that minimal proline based models show enhanced binding selectivity towards phenol in CDCl(3), relative to other similarly protected amino acid residues. Cooperative binding effects appear to play a role, with sarcosine models affording binding constants to phenol intermediate to those obtained from proline models and other amino acid models. The mechanism for binding, based on DFT calculations and the application of Hunter's molecular recognition toolbox model, cannot be solely attributed to hydrogen bond strength, and appears to be mediated through C-H-pi bonds and the rotational freedom of the amide substrate. PMID- 18421392 TI - Efficient and flexible synthesis of chiral gamma- and delta-lactones. AB - An efficient and highly flexible synthesis for chiral gamma- and delta-lactones with high enantiomeric purity is described (>99% ee and 57-87% overall yield). The protocol involves alkylation of chiral 1,2-oxiranes with terminally unsaturated Grignard reagents. Subsequent oxidative degradation (OsO(4)-Oxone) of the terminal double bond from chiral alk-1-en-5-ols and alk-1-en-6-ols affords 4- or 5-hydroxy acids and gamma- and delta-lactones after acidic workup. The flexibility and efficiency of the protocol is illustrated by the synthesis of several alkanolides and alkenolides, hydroxy fatty acids and dihydroisocoumarins. PMID- 18421393 TI - On the effect of donor and acceptor substituents on the behaviour of light-driven rotary molecular motors. AB - Light-driven rotary molecular motors based on overcrowded alkenes can be substituted with electron-donating and electron-withdrawing substituents (R = OMe, Cl and CN) in direct conjugation with the central double bond (the axis of rotation) without having a significant influence on the rate-limiting, thermal isomerisation step of their rotary cycle. This indicates that in this system, it is predominantly steric factors that determine the barrier to the thermal helix inversion. In contrast, the quantum yield and photoequilibria in the photochemical step were found to be quite sensitive to the combination of substituent and solvent employed. PMID- 18421394 TI - Photoelectrochemical evaluation of pH effect on hole transport through triplex forming DNA immobilized on a gold electrode. AB - We characterized pH effect on hole transport through DNA duplexes possessing a partial triplex-forming region. Direct electrochemical measurement of the current response of photosensitizer-tethered DNA immobilized on a gold electrode revealed that the partial triplex formation under acidic conditions suppressed photocurrent due to hole transport, while dissociation of the triplex into the duplex as occurred upon increasing pH values recovered the photocurrent efficiency. Reversible conversion between duplex and triplex induced upon cyclic alternation of pH values resulted in a rise and fall of photocurrent responses, indicating that pH change may feature in the switching function of hole transport in DNA. These electrochemical behaviors could be correlated to the results obtained in long-range photo-oxidative DNA cleavage experiments, in which DNA cleavage at the hole trapping site beyond the triplex region was significantly suppressed under triplex-forming acidic conditions. PMID- 18421395 TI - Aminolysis of 4-nitrophenyl phenyl carbonate and thionocarbonate: effects of amine nature and modification of electrophilic center from C[double bond]O to C[double bond]S on reactivity and mechanism. AB - A kinetic study is reported for the reactions of 4-nitrophenyl phenyl carbonate (5) and thionocarbonate (6) with a series of alicyclic secondary amines in 80 mol% H(2)O-20 mol% DMSO at 25.0 +/- 0.1 degrees C. The plots of k(obsd) vs. amine concentration are linear for the reactions of 5. On the contrary, the plots for the corresponding reactions of 6 curve upward as a function of increasing amine concentration, indicating that the reactions proceed through two intermediates (i.e., a zwitterionic tetrahedral intermediate T(+/-) and its deprotonated form T(-)). The Bronsted-type plot for 5 the reactions of with secondary amines exhibits a downward curvature, i.e., the slope decreases from 0.98 to 0.26 as the pK(a) of the conjugate acid of amines increases, implying that the reactions proceed through T(+/-) with a change in the rate-determining step (RDS). The k(N) values are larger for the reactions of with secondary amines than for those with primary amines of similar basicity. Dissection of k(N) values for the reactions of 5 into the microscopic rate constants (i.e., k(1) and k(2)/k(-1) ratio) has revealed that k(1) is larger for the reactions with secondary amines than for those with isobasic primary amines, while the k(2)/k(-1) ratio is nearly identical. On the other hand, for reactions of 6, secondary amines exhibit larger k(1) values but smaller k(2)/k(-1) ratios than primary amines. The current study has shown that the reactivity and reaction mechanism are strongly influenced by the nature of amines (primary vs. secondary amines) and electrophilic centers (C[double bond]O vs. C[double bond]S). PMID- 18421396 TI - An oxidatively-activated safety catch linker for solid phase synthesis. AB - A N-benzyl-4-amino-2,2-dimethylbutanoic acid-based system has been developed as a new oxidatively activated safety catch linker for reaction monitoring and optimisation on solid support. The CAN promoted oxidative debenzylation of the tertiary N-benzylamine moiety, followed by concomitant cyclisation and release of alcohols and amines has been demonstrated both in solution phase model studies and on the solid phase. The linker system has been applied to the solid phase synthesis of a collection of phenol derivatives, and to the demonstration of the attachment and release of a chiral auxiliary from a solid support. PMID- 18421397 TI - Stereoselective NaN3-catalyzed halonitroaldol-type reaction of azetidine-2,3 diones in aqueous media. AB - Azetidine-2,3-diones (alpha-oxo-beta-lactams) and bromonitromethane undergo coupling in aqueous media in the presence of catalytic amounts of sodium azide. The stereoselectivity of the process was generally good, proceeding with reasonable anti : syn ratios under substrate control. On this basis, a simple and fast protocol for the synthesis of the potentially bioactive 3-substituted 3 hydroxy-beta-lactam moiety has been developed. Besides, 2-azetidinone-tethered 1 halo-1-nitroalkan-2-ols are quite useful building blocks; for example, reactions of the above nitrobromohydrins provided spiranic and fused bicyclic-beta-lactams. PMID- 18421398 TI - Stereochemistry of 1,2-elimination reactions at the E2-E1cB interface--tert-butyl 3-tosyloxybutanoate and its thioester. AB - Experimental data on the stereoselectivity of base-catalyzed 1,2-elimination reactions that produce conjugated carbonyl compounds are scarce in spite of the importance of these reactions in organic and biochemistry. As part of a comprehensive study in this area, we have synthesized stereospecifically deuterated beta-tosyloxybutanoate esters and thioesters and studied the stereoselectivity of their elimination reactions under non-ion pairing conditions. With the availability of both the (2R*,3R*) and (2R*,3S*) diastereomers the innate stereoselectivity could be determined unambiguously. (1)H and (2)H NMR data show that these substrates produce 5-6% syn elimination, the usual amount for acyclic substrates undergoing E2 reactions. Contrary to earlier suggestions, activation by a carbonyl group has virtually no influence upon the stereoselectivity. Elimination of the (2R*,3R*) diastereomer of the beta tosyloxyester and thioester produces 21-25% of the (Z)-alkene, much more than observed with a poorer beta-nucleofuge. A relatively large amount of (Z)-alkene product seems to be a good marker for an E2 pathway, in which the transition state is E1cB-like, rather than an E1cB(irrev) mechanism. Syn KIE values were higher than those for anti elimination for the esters as well as the thioesters. Experimental challenges to the synthesis of stereospecifically-deuterated beta tosyloxyesters are discussed. PMID- 18421399 TI - Synthesis and evaluation of novel pyridine based PLG tripeptidomimetics. AB - Analogues of the pyridine based PLG (Pro-Leu-Gly-NH(2)) peptidomimetic were synthesized and evaluated as dopamine modulating agents. Modifications in the position corresponding to the leucine side chain in PLG afforded derivatives , and , substituted with H, Me and Bn instead of the isobutyl group, respectively. Changes in the proline residue produced derivative , substituted with a symmetrical piperidine ring instead of the pyrrolidine ring and , in which the pyrrolidine ring is connected to the pyridine ring via a hydroxymethyl group instead of a keto function. The peptidomimetics were tested for their ability to enhance the maximal effect of N-propylapomorphine (NPA) at dopamine D2 receptors in the functional cell-based R-SAT assay. Compounds , , and , produced a statistically significant increase in the maximal NPA response at 10 nM (117 +/- 6%, 118 +/- 6%, and 116 +/- 3%, respectively), which is similar to the effect of PLG in this assay, whereas was able to potentiate the response to a similar extent at 1 nM concentration (115 +/- 5%). All derivatives produced a bell-shaped dose-response curve and none of the compounds were active at the D2 receptor alone, which indicates that the mechanism behind the activity of both the pyridine based mimetics and PLG is the same. Interestingly, l-Pro-d-Leu-Gly-NH(2) was found to be more potent than PLG and produced a 119 +/- 1% increase in the NPA response at 1 nM. PMID- 18421400 TI - Asymmetric synthesis of vicinal amino alcohols: xestoaminol C, sphinganine and sphingosine. AB - The highly diastereoselective anti-aminohydroxylation of alpha,beta-unsaturated esters, via conjugate addition of lithium (S)-N-benzyl-N-(alpha methylbenzyl)amide and subsequent in situ enolate oxidation with (+) (camphorsulfonyl)oxaziridine, has been used as the key step in the asymmetric synthesis of N,O-diacetyl xestoaminol C (41% yield over 8 steps), N,O,O-triacetyl sphinganine (30% yield over 8 steps) and N,O,O-triacetyl sphingosine (30% yield over 7 steps). PMID- 18421401 TI - Asymmetric synthesis of N,O,O,O-tetra-acetyl d-lyxo-phytosphingosine, jaspine B (pachastrissamine), 2-epi-jaspine B, and deoxoprosophylline via lithium amide conjugate addition. AB - The highly diastereoselective anti-aminohydroxylation of (E)-gamma-tri-iso propylsilyloxy-alpha,beta-unsaturated esters, via conjugate addition of lithium (S)-N-benzyl-N-(alpha-methylbenzyl)amide and subsequent in situ enolate oxidation with (+)-(camphorsulfonyl)oxaziridine, has been used as the key step in the asymmetric synthesis of N,O,O,O-tetra-acetyl d-lyxo-phytosphingosine (20% yield over 7 steps), the anhydrophytosphingosine jaspine B (10% yield over 9 steps), 2 epi-jaspine B (14% yield over 9 steps), and the Prosopis alkaloid deoxoprosophylline (26% yield over 7 steps). PMID- 18421402 TI - Pyrrolo-dC oligonucleotides bearing alkynyl side chains with terminal triple bonds: synthesis, base pairing and fluorescent dye conjugates prepared by the azide-alkyne "click" reaction. AB - 5-(Octa-1,7-diynyl)-2'-deoxyuridine was converted into the furano-dU derivative 7 by copper-catalyzed cyclization; the pyrolodC-derivative 3 was formed upon ammonolysis. The bicyclic nucleosides 3 and 7 as well as the corresponding non cyclic precursors 4 and 6 all containing terminal C[triple bond]C bonds were conjugated with the non-fluorescent 3-azido-7-hydroxycoumarin 5 employing the copper(I)-catalyzed Huisgen-Sharpless-Meldal cycloaddition "click reaction". Strongly fluorescent 1H-1,2,3-triazole conjugates (30-33) are formed incorporating two fluorescent reporters-the pyrdC nucleoside and the coumarin moiety. Oligonucleotides incorporating 6-alkynyl and 6-alkyl 7H-pyrrolo[2,3 d]pyrimidin-2(3H)-one nucleosides (3 and 2f) have been prepared by solid-phase synthesis using the phosphoramidite building blocks 10 and 13 ; the pyrrolo-dC oligonucleotides are formed during ammonia treatment. The duplex stability of oligonucleotides containing 3 and related derivatives was studied. Oligonucleotides with terminal triple bonded nucleosides such as 3 are more stabilizing than those lacking a side chain with terminal unsaturation; open chain derivatives (4) are even more efficient. The click reaction was also performed on oligonucleotides containing the pyrdC-derivative and the fluorescence properties of nucleosides, oligonucleotides and their coumarin conjugates were studied. PMID- 18421403 TI - Using blood plasma for monitoring organochlorine contaminants in juvenile white sturgeon, Acipenser transmontanus, from the lower Columbia River. AB - Organochlorine (OC) pesticide concentrations in blood plasma samples from 88 juvenile white sturgeon collected from the lower Columbia River were measured and compared to plasma sex steroid and OC tissue levels previously measured in corresponding fish. Significant squared correlation coefficients between summation operator DDT concentrations in sturgeon plasma and gonads and livers were 0.37 and 0.32, respectively. Significant negative correlations between plasma testosterone concentration and plasma Sigma DDT concentration in male fish (r(2)=0.26), plasma 17beta estradiol concentration and plasma Sigma DDT concentration in female fish (r(2)=0.38) and condition factor and plasma Sigma DDT concentration in all fish were found (r(2)=0.17). These results suggest that blood plasma may be a suitable nondestructive method for monitoring adult sturgeon population for persistent OC contaminants. PMID- 18421404 TI - Phytofiltration of copper from contaminated water: growth response, copper uptake and lignin content in Elsholtzia splendens and Elsholtzia argyi. AB - Phytofiltration of Cu from water by Elsholtzia plants were investigated hydroponically. Both E. splendens and E. argyi could effectively clean up lower Cu contaminated water, probably attributed to their extremely high surface area of roots. The removal rate of Cu was concentration-dependent and showed a monotonic decline with time. At higher external Cu concentration, the growth of E. argyi has been considerably inhibited while E. splendens still grew normally. E. splendens has the greater capacity to absorb Cu to roots from water and translocate Cu from roots to shoots. Lignin in roots probably has no significant effect on immobilizing Cu. PMID- 18421405 TI - The Fellows' Pelvic Research Network (FPRN). PMID- 18421406 TI - Moderate weight loss in obese women with urinary incontinence: a prospective longitudinal study. AB - This study assessed the effect of moderate weight loss in obese women with urodynamically proven urinary incontinence using the International Consultation on Incontinence recommended outcome measures. Sixty-four incontinent women were offered a weight reduction programme with a target loss of 5-10%. This included a low-calorie diet and exercise. An anti-obesity drug (Orlistat) was offered to those who failed to achieve their target. Forty-two (65%) achieved the target weight loss and had significant reduction in body mass index and girth. Weight loss was associated with significant reduction in pad test loss (median difference, 19 g; 95% confidence interval, 13-28 g; p < 0.001). There was also a clinical and statistically significant improvement in quality of life measures. These results suggest that weight reduction of 5% of initial body weight can improve urinary incontinence severity and its effects on quality of life in obese women. PMID- 18421407 TI - Physiological effects of human muscle-derived stem cell implantation on urethral smooth muscle function. AB - The physiological effects of human muscle-derived stem cell (MDSC) implantation on urethral smooth muscle function were investigated in pudendal nerve-transected nude rats with human MDSC (TM) or saline (TS) injection into the proximal urethra compared with sham-operated, saline-injected nude rats (SS). Leak point pressure (LPP) before and after hexamethonium application, which can block autonomic efferent nerves, and proximal urethral contractile responses to carbachol and phenylephrine in muscle strip study were examined 6 weeks after the implantation. There was no significant difference between the LPPs in SS and TM. Following hexamethonium application, the LPP in TM was, however, significantly decreased compared with SS. The contractile responses to phenylephrine, but not to carbachol, in TM were significantly increased compared with SS and TS. These results suggest that the restorative effects of MDSCs are mediated by autonomic nerves and that increased sensitivity of alpha(1)-adrenoceptors may be related to restore the deficient urethral function. PMID- 18421408 TI - Metabolic failure precedes intracranial pressure rises in traumatic brain injury: a microdialysis study. AB - BACKGROUND: Cerebral microdialysis (MD) is able to detect markers of tissue damage and cerebral ischaemia and can be used to monitor the biochemical changes subsequent to head injury. In this prospective, observational study we analysed the correlation between microdialysis markers of metabolic impairment and intracranial pressure (ICP) and investigated whether changes in biomarker concentration precede rises in ICP. METHODS: MD and ICP monitoring was carried out in twenty-five patients with severe TBI in Neurointensive care. MD samples were analysed hourly for lactate:pyruvate (LP) ratio, glutamate and glycerol. Abnormal values of microdialysis variables in presence of normal ICP were used to calculate the risk of intracranial hypertension developing within the next 3 h. FINDINGS: An LP ratio >25 and glycerol >100 micromol/L, but not glutamate >12 micromol/L, were associated with significantly higher risk of imminent intracranial hypertension (odds ratio: 9.8, CI 5.8-16.1; 2.2, CI 1.6-3.8; 1.7, CI 0.6-3, respectively). An abnormal LP ratio could predict an ICP rise above normal levels in 89% of cases, whereas glycerol and glutamate had a poorer predictive value. CONCLUSIONS: Changes in the compound concentrations in microdialysate are a useful tool to describe molecular events triggered by TBI. These changes can occur before the onset of intracranial hypertension, suggesting that biochemical impairment can be present before low cerebral perfusion pressure is detectable. This early warning could be exploited to expand the window for therapeutic intervention. PMID- 18421409 TI - Inflation-deflation method for endoscopic evacuation of intracerebral haematoma. AB - BACKGROUND: Endoscopic evacuation of intracerebral haematoma (ICH) has the advantage of being less invasive than craniotomy, but limited visualisation and difficulties in haemostasis are still a concern. The collapse of the haematoma cavity limits the visualisation of the surgical field. Inflation of the haematoma cavity with saline irrigation improves visualisation and facilitates accurate intra-operative orientation. METHOD: A unique technique, the cavity inflation deflation method can help in exploring the residual haematoma and accessing the bleeding points. We also developed a combined irrigation-coagulation suction tube that concentrates the capabilities of suction, irrigation and monopolar coagulation. FINDINGS: The use of this multifunctional dedicated instrument and its application in the cavity inflation-deflation method allows for easy identification of residual haematoma and bleeding vessels. Secure haemostasis can also be accomplished under clear visualisation. No surgical complications and rebleeding occurred in any patient following the procedure. Our results show that the median haematoma evacuation rate was 99% and the surgical outcome was satisfactory. CONCLUSIONS: The inflation-deflation method using a combined irrigation-coagulation suction tube can facilitate optimal evacuation of ICH with secure haemostasis. Although further accumulation of patients and careful analyses are needed to be known whether this procedure improves the clinical outcomes in the patients, the preliminary results of its application have been promising. PMID- 18421410 TI - Surgical treatment of spinal dural arterio-venous fistulae: long-term results and analysis of prognostic factors. AB - BACKGROUND: Spinal dural arterio-venous fistulae (SDAVF) are slow-flow extramedullary vascular lesions which account for 75-80% of all spinal vascular malformations. At present there is no agreed view with regard to the best therapeutic option being surgical or endovascular, and several reports favour one or other form of management. This is so because of lack of consistent literature, as well as knowledge, concerning the long-term clinical outcome of the patients. The objective of this study is to retrospectively analyse the results obtained with patients operated for a SDAVF at the Department of Neurosurgery of Verona during a 15-year period and to evaluate possible prognostic factors related to neurological outcome. PATIENTS AND METHODS: Between January 1987 and May 2002, 29 patients with SDAVF were operated at the Department of Neurosurgery of Verona. For 25 of these patients we were able to obtain a clinical follow-up using telephone interviews. The patients were evaluated with the Aminoff and Logue's scale and subsequently stratified into three classes of disability. An overall score (gait and micturition, G + M) of 0-3 indicates a mild disability, a score between 4 and 5 indicates a moderate disability and a score between 6 and 8 a severe disability. All patients underwent surgical treatment which was mainly the first therapeutic option. Following surgery, the patients were re-evaluated with the same neurological scale. We also investigated with statistical analysis the possible impact on clinical outcome of the major clinical, neuroradiological and surgical variables. RESULTS: The epidemiological, clinical, radiological and pathological features of our group of patients are very similar to those previously described in the literature. For 10 patients surgery consisted simply of the interruption of the intradural arterialized draining vein (with or without closure of the small extradural arterial afferents), whereas in the remaining 15 patients coagulation or excision of the fistulous dura was also accomplished. At the last follow-up (mean 7.3 years; in 19 patients longer than 5 years), 10 patients had improved (40%), 11 were stable (44%) and 4 had deteriorated (16%). We determined that only the pre-operative neurological status, described by the G value in the Aminoff and Logue's scale and the class of disability, had an impact on clinical outcome. CONCLUSIONS: This retrospective study confirms that the surgical treatment results of SDAVF are satisfactory even if evaluated after many years. Given these results, and in accordance with the majority of the literature, we concur that surgery should be the first choice treatment for these spinal vascular lesions in order to avoid a dangerous delay and consequently further neurological deterioration. In our group of patients the only prognostic factor statistically related to clinical outcome was the pre-treatment neurological status, particularly the grade of paraparesis and the class of disability. PMID- 18421411 TI - Visual outcome after surgery for cavernous sinus meningioma. AB - BACKGROUND: Extension of cavernous sinus meningiomas can compromise vision by compressing the optic nerves and chiasm. Surgical tumour removal aims to protect vision in the long-term. However, the risks of surgery include transient or permanent damage to the anterior visual pathways. This study aims to 1) analyse the visual status in unilateral cavernous sinus meningioma with extra-cavernous extension, before and after removal of the extra-cavernous portion, without any adjuvant treatment with radiotherapy, 2) identify pre-surgical and early post surgical prognostic factors for long-term visual outcome and 3) compare these results to previous studies. METHODS: This is a retrospective study of 30 consecutive patients who underwent surgery between 1989 and 2004. Visual acuity, visual fields, and fundi were evaluated before surgery and during the mean follow up period of 2 years. FINDINGS: Total visual loss occurred in the ipsilateral eye following surgery in 10% of patients. There was improvement in vision in 23%, no significant change in 27% and worsening in 50% of patients. Pre-surgical visual acuity was not predictive of final visual outcome, but initial optic disc pallor was a poor prognostic factor. In pre-operatively normal contralateral eyes, 10% developed a mild visual deficit (visual acuity = 20/32 or better, and visual field defect 0.05), at the same times of study. Mg(2+)-ATPase activities remained unaltered in both groups of mothers and newborns. Na(+), K(+)-ATPase activity was similarly lower in the CB of neonates than those of their mothers, pre-delivery. Our results suggest that: (a) during a normal vaginal delivery process, the low TAS and the increased levels of catecholamines may increase Na(+), K(+)-ATPase activity, post-delivery; (b) the low enzyme activities evaluated in mothers pre delivery may be due to the high estrogen levels and those in newborns due to perinatal immaturity. PMID- 18421472 TI - Bladder cooling reflex and external urethral sphincter activity in the anesthetized and awake guinea pig. AB - A spinal bladder cooling reflex, triggered by cold receptors of transient receptor potential melastatin type in the bladder wall, has been identified in several mammals, including man. This reflex and its influence on the external urethral sphincter were further characterized in the urethane anesthetized and awake guinea pigs. A total of 214 bladder infusions were performed in the 12 animals. Compared to controls, cold fluid induced a significant decrease in the threshold volume for reflex bladder contractions (median 82%, p < 0.01). Menthol induced a further decrease (median 50%), signifying a bladder cooling reflex. Detrusor-sphincter activities were dyssynergic during voidings triggered by cold or menthol infusions but were coordinated during control infusions. The bladder cooling reflex was suppressed and the sphincter activity synergic following cold infusions in the awake state. Thus, the bladder cooling reflex is under the active descending inhibitory control in intact, awake animals. PMID- 18421471 TI - Cardiac thin filament regulation. AB - Myocardial contraction is initiated upon the release of calcium into the cytosol from the sarcoplasmic reticulum following membrane depolarization. The fundamental physiological role of the heart is to pump an amount blood that is determined by the prevailing requirements of the body. The physiological control systems employed to accomplish this task include regulation of heart rate, the amount of calcium release, and the response of the cardiac myofilaments to activator calcium ions. Thin filament activation and relaxation dynamics has emerged as a pivotal regulatory system tuning myofilament function to the beat-to beat regulation of cardiac output. Maladaptation of thin filament dynamics, in addition to dysfunctional calcium cycling, is now recognized as an important cellular mechanism causing reduced cardiac pump function in a variety of cardiac diseases. Here, we review current knowledge regarding protein-protein interactions involved in the dynamics of thin filament activation and relaxation and the regulation of these processes by protein kinase-mediated phosphorylation. PMID- 18421473 TI - Functional consequences of leucine and tyrosine mutations in the dual pore motifs of the yeast K(+) channel, Tok1p. AB - Tandem pore-loop potassium channels differ from the majority of K(+) channels in that a single polypeptide chain carries two K(+)-specific segments (P) each sandwiched between two transmembrane helices (M) to form an MP(1)M-MP(2)M series. Two of these peptide molecules assemble to form one functional potassium channel, which is expected to have biaxial symmetry (commonly described as asymmetric) due to independent mutation in the two MPM units. The resulting intrinsic asymmetry is exaggerated in fungal 2P channels, especially in Tok1p of Saccharomyces, by the N-terminal presence of four more transmembrane helices. Functional implications of such structural asymmetry have been investigated via mutagenesis of residues (L290 in P(1) and Y424 in P(2)) that are believed to provide the outermost ring of carbonyl oxygen atoms for coordination with potassium ions. Both complementary mutations (L290Y and Y424L) yield functional potassium channels having quasi-normal conductance when expressed in Saccharomyces itself, but the P(1) mutation (only) accelerates channel opening about threefold in response to depolarizing voltage shifts. The more pronounced effect at P(1) than at P(2) appears paradoxical in relation to evolution, because a comparison of fungal Tok1p sequences (from 28 ascomycetes) shows the filter sequence of P(2) (overwhelmingly TIGYGD) to be much stabler than that of P(1) (mostly TIGLGD). Profound functional asymmetry is revealed by the fact that combining mutations (L290Y + Y424L)-which inverts the order of residues from the wild-type channel reduces the expressed channel conductance by a large factor (20-fold, cf. 95th percentile. A history of asthma was reported by 31.4% (74/236); no subject was symptomatic on the night of the study. We found no increased risk for polysomnographically diagnosed OSA for asthmatics. To the contrary, by logistic regression analysis, a parent/guardian report of asthma decreased the odds of having OSA by 34% (p = 0.027), controlling for individual and socioeconomic factors and assessment results. Polysomnographic (PSG) differences between asthmatic and non-asthmatic children were found in only the arousal index (11.0 vs.9.3 +/- 6.5/h, p = 0.099) and total sleep time (337.1 +/- 64.3 vs. 347 +/- 65.2 min, p = 0.1) In a referral-based group of predominantly AA inner-city snoring children, asymptomatic asthma decreased the likelihood of OSA. PMID- 18421492 TI - Metagenomics and the niche concept. AB - The metagenomics approach has revolutionised the fields of bacterial diversity, ecology and evolution, as well as derived applications like bioremediation and obtaining bioproducts. A further associated conceptual change has also occurred since in the metagenomics methodology the species is no longer the unit of study, but rather partial genome arrangements or even isolated genes. In spite of this, concepts coming from ecological and evolutionary fields traditionally centred on the species, like the concept of niche, are still being applied without further revision. A reformulation of the niche concept is necessary to deal with the new operative and epistemological challenges posed by the metagenomics approach. To contribute to this end, I review past and present uses of the niche concept in ecology and in microbiological studies, showing that a new, updated definition need to be used in the context of the metagenomics. Finally, I give some insights into a more adequate conceptual background for the utilisation of the niche concept in metagenomic studies. In particular, I raise the necessity of including the microbial genetic background as another variable into the niche space. PMID- 18421493 TI - Serum levels of TRAP5b, a new bone resorption marker unaffected by renal dysfunction, as a useful marker of cortical bone loss in hemodialysis patients. AB - Tartrate-resistant acid phosphatase (TRAP) 5b is a new marker of bone resorption that is unaffected by renal dysfunction. The significance of TRAP5b was assessed in hemodialysis (HD) patients. Serum concentrations of TRAP5b and cross-linked N telopeptide of type I collagen (NTX) were determined as bone resorption markers, and those of bone alkaline phosphatase (BAP) and intact osteocalcin (OC) were measured as bone formation markers in 58 HD patients. Bone mineral density (BMD) was measured by dual-energy X-ray absorptiometry twice in the distal third of the radius, with a 2-year interval between measurements. Serum TRAP5b correlated significantly with BAP, intact OC, intact parathyroid hormone (PTH), and especially serum NTX. TRAP5b, NTX, BAP, and intact OC all correlated significantly with BMD at the time of the second measurement; and TRAP5b, NTX, and intact OC, but not BAP and intact PTH, correlated significantly with the annual change in BMD during the 2-year period. Among the bone markers, patients in the highest tertile for serum TRAP5b and intact OC showed the fastest rate of cortical bone loss. The sensitivity and specificity for detection of rapid bone loss were 57.9% and 76.9%, respectively, for serum TRAP5b. Measurement of serum TRAP5b, as well as intact OC, may be a clinically relevant assay for estimation of bone metabolic status in HD patients, although serum intact OC accumulates in uremic serum. PMID- 18421495 TI - Differential toxicity to Cd, Pb, and Cu in dragonfly larvae (Insecta: Odonata). AB - Odonate larvae are important organisms in aquatic ecosystems but have been rarely studied in laboratory toxicity tests. Only a few previous studies have been conducted on odonates and their responses to heavy metals. We exposed two species of libellulid larvae (Anisoptera: Libellulidae) to equimolar concentrations of cadmium, lead, or copper in 7-day survival tests. Larvae were tolerant of high concentrations of cadmium and lead, as no significant decrease in survival was observed at exposures as high as 0.893 and 2.232 mM, respectively. In contrast, larvae were more sensitive to copper exposure, demonstrating significantly decreased survival to exposures as low as 2.360 microM. In whole animal samples, larvae accumulated very high concentrations (>1000 microg/g dry weight) of all three metals in an exposure-related manner. Much of this accumulation could probably be attributed to adsorption or accumulation of metal within the exoskeleton, because odonate larvae are known to sequester metals into this material. Our results were generally consistent with previous observations indicating that odonates are tolerant to metal exposures, even in comparison with other aquatic invertebrates. However, there are few studies that have used odonates in toxicity tests and compared these organisms to other aquatic life. Based on their abundance and their simple requirements in the laboratory, we believe that odonate larvae can be useful toxicological model organisms. PMID- 18421494 TI - Mutations at arginine 352 alter the pore architecture of CFTR. AB - Arginine 352 (R352) in the sixth transmembrane domain of the cystic fibrosis transmembrane conductance regulator (CFTR) previously was reported to form an anion/cation selectivity filter and to provide positive charge in the intracellular vestibule. However, mutations at this site have nonspecific effects, such as inducing susceptibility of endogenous cysteines to chemical modification. We hypothesized that R352 stabilizes channel structure and that charge-destroying mutations at this site disrupt pore architecture, with multiple consequences. We tested the effects of mutations at R352 on conductance, anion selectivity and block by the sulfonylurea drug glipizide, using recordings of wild-type and mutant channels. Charge-altering mutations at R352 destabilized the open state and altered both selectivity and block. In contrast, R352K-CFTR was similar to wild-type. Full conductance state amplitude was similar to that of wild-type CFTR in all mutants except R352E, suggesting that R352 does not itself form an anion coordination site. In an attempt to identify an acidic residue that may interact with R352, we found that permeation properties were similarly affected by charge-reversing mutations at D993. Wild-type-like properties were rescued in R352E/D993R-CFTR, suggesting that R352 and D993 in the wild-type channel may interact to stabilize pore architecture. Finally, R352A-CFTR was sensitive to modification by externally applied MTSEA+, while wild-type and R352E/D993R-CFTR were not. These data suggest that R352 plays an important structural role in CFTR, perhaps reflecting its involvement in forming a salt bridge with residue D993. PMID- 18421496 TI - Species differences in the sensitivity of avian embryos to methylmercury. AB - We injected doses of methylmercury into the air cells of eggs of 26 species of birds and examined the dose-response curves of embryo survival. For 23 species we had adequate data to calculate the median lethal concentration (LC(50)). Based on the dose-response curves and LC(50)s, we ranked species according to their sensitivity to injected methylmercury. Although the previously published embryotoxic threshold of mercury in game farm mallards (Anas platyrhynchos) has been used as a default value to protect wild species of birds, we found that, relative to other species, mallard embryos are not very sensitive to injected methylmercury; their LC(50 )was 1.79 microg/g mercury on a wet-weight basis. Other species we categorized as also exhibiting relatively low sensitivity to injected methylmercury (their LC(50)s were 1 microg/g mercury or higher) were the hooded merganser (Lophodytes cucullatus), lesser scaup (Aythya affinis), Canada goose (Branta canadensis), double-crested cormorant (Phalacrocorax auritus), and laughing gull (Larus atricilla). Species we categorized as having medium sensitivity (their LC(50)s were greater than 0.25 microg/g mercury but less than 1 microg/g mercury) were the clapper rail (Rallus longirostris), sandhill crane (Grus canadensis), ring-necked pheasant (Phasianus colchicus), chicken (Gallus gallus), common grackle (Quiscalus quiscula), tree swallow (Tachycineta bicolor), herring gull (Larus argentatus), common tern (Sterna hirundo), royal tern (Sterna maxima), Caspian tern (Sterna caspia), great egret (Ardea alba), brown pelican (Pelecanus occidentalis), and anhinga (Anhinga anhinga). Species we categorized as exhibiting high sensitivity (their LC(50)s were less than 0.25 microg/g mercury) were the American kestrel (Falco sparverius), osprey (Pandion haliaetus), white ibis (Eudocimus albus), snowy egret (Egretta thula), and tri colored heron (Egretta tricolor). For mallards, chickens, and ring-necked pheasants (all species for which we could compare the toxicity of our injected methylmercury with that of published reports where methylmercury was fed to breeding adults and was deposited into the egg by the mother), we found the injected mercury to be more toxic than the same amount of mercury deposited naturally by the mother. The rank order of sensitivity of these same three species to methylmercury was, however, the same whether the methylmercury was injected or maternally deposited in the egg (i.e., the ring-necked pheasant was more sensitive than the chicken, which was more sensitive than the mallard). It is important to note that the dose-response curves and LC(50)s derived from our egg injections are useful for ranking the sensitivities of various species but are not identical to the LC(50)s that would be observed if the mother bird had put the same concentrations of mercury into her eggs; the LC(50)s of maternally deposited methylmercury would be higher. PMID- 18421497 TI - Left unilateral breast autoinflation and intraprosthetic collection of sterile pus: an unusual operative finding of silicone gel bleed with silicone lymphadenitis. AB - Implant rupture is a known complication after augmentation mammoplasty. Saline implant failure may present with few diagnosable problems due to the loss of volume. On the other hand, intracapsular silicone gel implant rupture or failure can be silent, asymptomatic, and difficult to diagnose in the absence of loss of breast volume. Silicone leak and migration to regional nodes has been reported but is uncommon and lymph node enlargement can be nonspecific in patients with intact prostheses. Causes, timing, and diagnosis of implant failure sites and mechanisms have been extensively studied and reported in the literature. Implant failure resulting in lymphadenopathy is uncommon. In this article autoinflation of the left breast with regional lymphadenopathy is presented. Upon exploration, the prosthesis was found to be filled with sterile pus, showing an inward movement of a thick creamy material in an otherwise macroscopically intact prosthesis. PMID- 18421498 TI - Context and systems: thinking more broadly about effectiveness in strategic environmental assessment in China. AB - China is an illustrative--and extreme--case of the difficulties of balancing the pursuit of economic, social, and environmental objectives. In 2003 it adopted a form of Strategic Environmental Assessment (SEA) for its plans and programs (referred to here as PEIA) with the aim of moving towards greater environmental sustainability. The literature has explored primarily the issue of methods and legal procedures. This research contributes to the analysis of PEIA through a different set of interpretative lens. Drawing on recent developments in the theory and practice of SEA, I propose a conceptualization of SEA effectiveness that combines direct and incremental impacts, and a need for context-specific systems as a way to focus on the relationship between assessment, planning, and their context, and thus maximize effectiveness. This framework underpins the analysis of China's experience, which I explore with the help of interview material and the literature. The result is an evaluation of the strengths and weaknesses of PEIA in terms of its purpose, assessment concept, process, and methods. The detailed analysis of six aspects of the context helps explain the origin of such shortcomings, and identify opportunities for its improvement. I conclude defining elements of a context-specific system for SEA that seeks to maximize the opportunity for incremental, as well as direct, effectiveness in China. PMID- 18421499 TI - Imaging-guided percutaneous radiofrequency ablation of adrenal metastases: preliminary results at a single institution with a single device. AB - The aim of this study was to show the feasibility, safety, imaging appearance, and short-term efficacy of image-guided percutaneous radiofrequency ablation (RFA) of adrenal metastases (AM). Seven imaging-guided percutaneous RFA treatments were performed in six patients (two men and four women; mean age, 67.2 years; range, 55-74 years) with six AM who were referred to our institution from 2003 to 2006. One patient was treated twice for recurrence after first treatment. The average diameter of the treated AM was 29 mm (range, 15-40 mm). In all patients, the diagnosis was obtained with CT current protocols in use at our institution and confirmed by pathology with an image-guided biopsy. No major complications occurred. In one patient shortly after initiation of the procedure, severe hypertension was noted; another patient developed post-RFA syndrome. In five of six lesions, there was no residual enhancement of the treated tumor. In one patient CT examination showed areas of residual enhancement of the tumor after treatment. Our preliminary results suggest that imaging-guided percutaneous RFA is effective for local control of AM, without major complications and with a low morbidity rate related to the procedure. Long-term follow-up will need to be performed and appropriate patient selection criteria will need to be determined in future randomized trials. PMID- 18421502 TI - Subcutaneous fissurotomy included a disguised and unappreciated anal spincter dilatation. PMID- 18421501 TI - Three-dimensional architecture of hair-bundle linkages revealed by electron microscopic tomography. AB - The senses of hearing and balance rest upon mechanoelectrical transduction by the hair bundles of hair cells in the inner ear. Located at the apical cellular surface, each hair bundle comprises several tens of stereocilia and a single kinocilium that are interconnected by extracellular proteinaceous links. Using electron-microscopic tomography of bullfrog saccular sensory epithelia, we examined the three-dimensional structures of basal links, kinociliary links, and tip links. We observed significant differences in the appearances and dimensions of these three structures and found two distinct populations of tip links suggestive of the involvement of different proteins, splice variants, or protein protein interactions. We noted auxiliary links connecting the upper portions of tip links to the taller stereocilia. Tip links and auxiliary links show a tendency to adopt a globular conformation when disconnected from the membrane surface. PMID- 18421503 TI - Be cautious with administrative data. PMID- 18421505 TI - Studying the metabolic activity of red bone marrow by means of FDG-PET: the need for a standardization. PMID- 18421506 TI - Establishing the diagnosis of Libman-Sacks endocarditis in systemic lupus erythematosus. AB - CASE: A 43-year-old female with systemic lupus erythematosus (SLE) was admitted with fever and shortness of breath 1 month after aortic valve replacement. A diagnostic workup including chemistries, complete blood count, blood cultures, chest x-ray, and 2-D echocardiogram was performed to determine the etiology of her symptoms and differentiate between acute bacterial endocarditis and Libman Sacks endocarditis. DISCUSSION: By utilizing Duke's criteria, antiphospholipid antibodies, and serial echocardiography, we were able to make a diagnosis of Libman-Sacks endocarditis. The patient was successfully treated for Libman-Sacks endocarditis and recovered uneventfully. CONCLUSION: This case highlights the challenges of making the correct diagnosis when 2 disease processes present with similar findings. PMID- 18421507 TI - Stop orders to reduce inappropriate urinary catheterization in hospitalized patients: a randomized controlled trial. AB - BACKGROUND: Hospitalized patients frequently have urinary catheters inserted for inappropriate reasons. This can lead to urinary tract infections and other complications. OBJECTIVE: To assess whether stop orders for indwelling urinary catheters reduces the duration of inappropriate urinary catheterization and the incidence of urinary tract infections. DESIGN: A randomized controlled trial was conducted in three tertiary-care hospitals in Ontario, Canada. Patients with indwelling urinary catheters were randomized to prewritten orders for the removal of urinary catheters if specified criteria were not present or to usual care. PARTICIPANTS: Six hundred ninety-two hospitalized patients admitted to hospital with indwelling urinary catheters inserted for < or = 48 h. MEASUREMENTS: The main outcomes included days of inappropriate indwelling catheter use, total days of catheter use, frequency of urinary tract infection, and catheter reinsertions. RESULTS: There were fewer days of inappropriate and total urinary catheter use in the stop-order group than in the usual care group (difference -1.69 [95% CI -1.23 to -2.15], P < 0.001 and -1.34 days, [95% CI, -0.64 to -2.05 days], P < 0.001, respectively). Urinary tract infections occurred in 19.0% of the stop-order group and 20.2% of the usual care group, relative risk 0.94 (95% CI, 0.66 to 1.33), P = 0.71. Catheter reinsertion occurred in 8.6% of the stop-order group and 7.0% in the usual care group, relative risk 1.23 (95% CI, 0.72 to 2.11), P = 0.45. CONCLUSIONS: Stop orders for urinary catheterization safely reduced duration of inappropriate urinary catheterization in hospitalized patients but did not reduce urinary tract infections. PMID- 18421509 TI - The ethics of information: absolute risk reduction and patient understanding of screening. AB - Some experts have argued that patients should routinely be told the specific magnitude and absolute probability of potential risks and benefits of screening tests. This position is motivated by the idea that framing risk information in ways that are less precise violates the ethical principle of respect for autonomy and its application in informed consent or shared decision-making. In this Perspective, we consider a number of problems with this view that have not been adequately addressed. The most important challenges stem from the danger that patients will misunderstand the information or have irrational responses to it. Any initiative in this area should take such factors into account and should consider carefully how to apply the ethical principles of respect for autonomy and beneficence. PMID- 18421508 TI - Improving HIV screening and receipt of results by nurse-initiated streamlined counseling and rapid testing. AB - BACKGROUND: HIV testing is cost-effective in unselected general medical populations, yet testing rates among those at risk remain low, even among those with regular primary care. HIV rapid testing is effective in many healthcare settings, but scant research has been done within primary care settings or within the US Department of Veteran's Affairs Healthcare System. OBJECTIVES: We evaluated three methods proven effective in other diseases/settings: nurse standing orders for testing, streamlined counseling, and HIV rapid testing. DESIGN: Randomized, controlled trial with three intervention models: model A (traditional counseling/testing); model B (nurse-initiated screening, traditional counseling/testing); model C (nurse-initiated screening, streamlined counseling/rapid testing). PARTICIPANTS: Two hundred fifty-one patients with primary/urgent care appointments in two VA clinics in the same city (one large urban hospital, one freestanding outpatient clinic in a high HIV prevalence area). MEASUREMENTS: Rates of HIV testing and receipt of results; sexual risk reduction; HIV knowledge improvement. RESULTS: Testing rates were 40.2% (model A), 84.5% (model B), and 89.3% (model C; p = <.01). Test result receipt rates were 14.6% (model A), 31.0% (model B), 79.8% (model C; all p = <.01). Sexual risk reduction and knowledge improvement did not differ significantly between counseling methods. CONCLUSIONS: Streamlined counseling with rapid testing significantly increased testing and receipt rates over current practice without changes in risk behavior or posttest knowledge. Increased testing and receipt of results could lead to earlier disease identification, increased treatment, and reduced morbidity/mortality. Policymakers should consider streamlined counseling/rapid testing when implementing routine HIV testing into primary/urgent care. PMID- 18421510 TI - Isolated renal relapse of sarcoidosis under low-dose glucocorticoid therapy. AB - Sarcoidosis is a multisystem disease of unknown etiology. Renal manifestation is rare and usually caused by hypercalcemia and nephrocalcinosis. Moreover, renal disease can occur as granulomatous interstitial nephritis (GIN), which is a histological diagnosis. We describe a case of sarcoidosis first presenting with multiple organ involvement including renal failure caused by severe GIN and subsequent remission on glucocorticoid therapy. After 18 months under low-dose prednisolone, the patient was readmitted with acute renal failure, histologically confirmed to be a relapse of renal sarcoidosis. Extrarenal manifestations of sarcoidosis were not present. Glucocorticoid dose was raised and kidney function again recovered significantly. Usual serologic markers of disease activity were not appropriate to indicate disease activity. Renal manifestation of sarcoidosis should be diagnosed by renal biopsy to guide therapy and probably requires larger glucocorticoid doses and prolonged treatment to prevent relapse. PMID- 18421511 TI - Effectiveness of the AUDIT-C as a screening test for alcohol misuse in three race/ethnic groups. AB - BACKGROUND: The Alcohol Use Disorders Identification Test-Consumption (AUDIT-C) is a brief validated screen for risky drinking and alcohol abuse and dependence (alcohol misuse). However, the AUDIT-C was validated in predominantly White populations, and its performance in different racial/ethnic groups is unclear. OBJECTIVE: To evaluate the validity of the AUDIT-C among primary care patients from the predominant racial/ethnic subgroups within the United States: White, African American, and Hispanic. DESIGN: Cross-sectional interview validation study. PARTICIPANTS: 1,292 outpatients from an academic family practice clinic in Texas (90% of randomly sampled eligible). MEASUREMENTS AND MAIN RESULTS: Race/ethnicity was self-reported. Areas under the receiver operating curve (AuROCs) evaluated overall AUDIT-C performance in the 3 racial/ethnic groups compared to diagnostic interviews for alcohol misuse. AUDIT-C sensitivities and specificities at recommended screening thresholds were compared across racial/ethnic groups. AuROCs were greater than 0.85 in all 3 groups, with no significant differences across racial/ethnic groups in men (P = .43) or women (P = .12). At previously recommended cut points, there were statistically significant differences by race in AUDIT-C sensitivities but not specificities. In women, the sensitivity was higher in Hispanic (85%) than in African-American (67%; P = .03) or White (70%; P = .04) women. In men, the sensitivity was higher in White (95%) than in African-American men (76%; P = .01), with no significant difference from Hispanic men (85%; P = .11). CONCLUSIONS: The overall performance of the AUDIT-C was excellent in all 3 racial/ethnic groups as reflected by high AuROCs. At recommended cut points, there were significant differences in the AUDIT-C's sensitivity but not in specificity across the 3 racial/ethnic groups. PMID- 18421530 TI - Serum S100B but not NSE levels are increased in morbidly obese individuals affected by obstructive sleep apnea-hypopnea syndrome. AB - BACKGROUND: Obstructive sleep apnea-hypopnea syndrome (OSAHS) is considered a comorbidity associated with morbid obesity, mainly because of the large neck circumference. Depending on its severity, OSAHS can interfere in many homeostasis systems, for example, the central nervous system (CNS). Neuron-specific enolase (NSE) and S100B protein derived from astrocytes are considered sensitive biochemical markers of cerebral injury. We evaluated serum S100B and NSE levels in this study with the aim of detecting possible cerebral injury as a consequence of OSAHS. METHODS: This was a transverse study with data from 25 morbidly obese patients with OSAHS. Blood samples were collected before and after polysomnography (PSG) to determine S100B and NSE protein levels. We also analyzed data evaluating depression and excessive daytime sleepiness. RESULTS: S100B levels were higher after [0.029 (0.010-0.199) mg/l] compared to before [0.010 (0.010-0.025) mg/l] on PSG (P = 0.002). S100B levels were expressed as means and IQ25-IQ75. NSE levels did not show significant differences before and after PSG. CONCLUSION: Our study shows a significant increase in S100B level after PSG compared to before. This suggests that there is a CNS astrocyte reaction because of possible cerebral hypoxemia in morbidly obese patients with OSAHS. PMID- 18421531 TI - Mechanical load and BMP signaling during tendon repair: a role for follistatin? AB - Healing of the rat Achilles tendon is sensitive to mechanical loading, and the callus strength is reduced by 3/4 after 14 days, if loading is prevented. Exogenous GDFs stimulate tendon healing. This response is influenced by loading: without loading, cartilage and bone formation is initiated. This implies BMP signaling is crucial during tendon healing and influenced by mechanical loading. We therefore asked if mechanical loading influences the gene expression of the BMP signaling system in intact and healing tendons, and how the BMP signaling system changes during healing. The genes were four BMPs (OP-1/BMP-7, GDF-5/CDMP 1/BMP-14, GDF-6/CDMP-2/BMP-13, and GDF-7/CDMP-3/BMP-12), two receptors (BMPR1b and BMPR2), and the antagonists follistatin and noggin. The Achilles tendon was transected in rats and left to heal. Half of the rats had one Achilles tendon unloaded by injection of Botox in the calf muscles. Ten tendons were analyzed before transection and for each of four time points. All genes except noggin were expressed at all time points, but followed different patterns during healing. Loading strongly decreased the expression of follistatin, which could lead to increased signaling. The BMP system appears involved in tendon maintenance and healing, and may respond to mechanical loading. PMID- 18421532 TI - Surgery can reduce the nonoperative care associated with an equinovarus foot deformity. AB - Equinovarus is the most common lower extremity deformity seen after a stroke. Despite its frequency, there are no specific guidelines in determining when surgery should be considered and for which patients it is appropriate. We evaluated the charges of nonsurgical and surgical treatments for equinovarus foot in 29 consecutive patients who underwent surgery for a unilateral equinovarus deformity after stroke. Twenty-six patients (seven males, 19 females) were available for followup. Mean patient age at the time of stroke was 48.2 years (range, 3-66 years). The average age at surgery was 54.7 years (range, 23-72 years), with a mean duration of nonsurgical treatment of 74.7 months. The minimum followup was 6 months following surgery (mean, 18.2 months; range, 6-48 months). Physical therapy accounted for 88% of nonoperative charges, with chemodenervation and orthotics accounting for 10% and 2%, respectively. Postoperatively, 19 patients were able to discontinue physical therapy compared with none preoperatively, and 17 discontinued orthotic use. Surgical correction of the equinovarus foot, in the appropriate patient, can decrease the use of nonoperative care for a patient who has had a stroke. We recommend surgery be considered earlier when an equinovarus deformity persists after the period of spontaneous neurologic recovery. LEVEL OF EVIDENCE: Level III, therapeutic study. PMID- 18421533 TI - Selected papers presented at the 2007 Meeting of the Musculoskeletal Tumor Society: editorial comment. PMID- 18421534 TI - 50 years ago in CORR: the iliac apophysis: an invaluable sign in the management of scoliosis Joseph C. Risser MD CORR 1958;11:111-119. PMID- 18421535 TI - Does the second-generation intercalary humeral spacer improve on the first? AB - Since publication of the results of a first-generation intercalary humeral spacer, a newer design has been available that addressed the weaknesses of the first. This study evaluated the hypothesis that the second-generation lap joint junction intercalary humeral spacer reduced complications compared with the original male-female taper design. We retrospectively reviewed the charts of 32 consecutive patients who had undergone placement of an intercalary humeral spacer. Twenty-one with the male-female taper (minimum followup 0 months, mean 19.2 months) were compared with 11 with the lap joint configuration (minimum followup 0 months, mean 20.3 months). Demographic, tumor, treatment, and radiographic variables were similar between groups. We observed a lower complication rate in the lap joint group (three of 11 versus 11 of 21). The most common complications in the male-female group, neuropraxia, periprosthetic fracture, and disengagement, were not seen in the lap joint group. Aseptic loosening was more frequent in the lap joint group. There were no differences in blood loss, operative time, or Musculoskeletal Tumor Society scores between groups. We noted improvement in Musculoskeletal Tumor Society scores from preoperatively to postoperatively in both groups. Use of these implants should be reserved for patients with limited life expectancy. LEVEL OF EVIDENCE: Level IV, therapeutic study. See the Guidelines for Authors for a complete description of levels of evidence. PMID- 18421536 TI - Attenuated Salmonella typhimurium with IL-2 gene reduces pulmonary metastases in murine osteosarcoma. AB - Historically, osteosarcoma has been a problematic metastatic disease, with 40-80% of patients developing pulmonary metastasis after primary tumor resection. Recent treatment advancements have reduced the occurrence of metastatic lesions to less than 30%. Using attenuated Salmonella typhimurium, we previously demonstrated regression in tumor burden in murine solid tumor and metastatic models. We established a murine model for metastatic osteosarcoma to determine the effect of treatment with a single oral dose of attenuated S. typhimurium with (SalpIL2) and without (Sal-NG) a gene for a truncated human interleukin-2. Female balb/c mice were administered 2 x 10(5) (ATCC K7M2) osteosarcoma cells via tail vein injection from culture and treated by oral gavage of Salmonella species 3 days later. Mice were harvested for splenic lymphocytes and tumor enumeration by intratracheal injection with India ink 21 days after injection. Treatment with attenuated SalpIL2 reduced pulmonary metastases in number and volume compared to saline controls. Furthermore, splenic natural killer cell populations were increased 93% with SalpIL2 and 114% with Sal-NG compared to nontreated groups. This pulmonary metastasis model demonstrates attenuated Salmonella typhimurium with human interleukin-2 reduced metastatic osteosarcoma in mice and confirm the need for further investigation into the immunologic properties of SalpIL2 as a possible treatment for metastatic osteosarcoma. PMID- 18421537 TI - FDG-PET imaging can diagnose periprosthetic infection of the hip. AB - A battery of diagnostic tests is often required to differentiate aseptic loosening from periprosthetic infection since the gold standard remains elusive. We designed a prospective study to determine the accuracy of fluorodeoxyglucose positron emission tomography (FDG-PET) imaging in diagnosing periprosthetic infection in a large multicenter setting. One hundred and thirteen patients with 127 painful hip prostheses were evaluated by FDG-PET. Images were considered positive for infection if PET demonstrated increased FDG activity at the bone prosthesis interface of the femoral component. A combination of preoperative tests, intraoperative findings, histopathology, and clinical followup constituted the gold standard for diagnosing infection. Among the 35 positive PET scans, 28 hips were confirmed infected according to our criteria for diagnosing periprosthetic infection. Of the 92 hip prostheses with negative FDG-PET findings, 87 were considered aseptic. The sensitivity, specificity, positive and negative predictive values for FDG-PET were 0.85 (28 of 33), 0.93 (87 of 94), 0.80 (28 of 35), and 0.95 (87 of 92), respectively. The overall accuracy of this novel noninvasive imaging modality reached 0.91 (115 of 127). Based on our results, FDG-PET appears a promising and accurate diagnostic tool for distinguishing septic from aseptic painful hip prostheses. LEVEL OF EVIDENCE: Level II, diagnostic study. See Guidelines for Authors for a complete description of levels of evidence. PMID- 18421538 TI - Polymicrobial prosthetic joint infections: risk factors and outcome. AB - Limited data are available regarding the risk factors and outcome of polymicrobial prosthetic joint infection (PJIs) when compared with monomicrobial PJI. Between January 1998 and November 2006, we retrospectively identified 34 of 174 prosthetic joint infections (19%) were polymicrobial. The 2-year cumulative probability of success of treating polymicrobial and monomicrobial PJIs was 63.8% and 72.8%, respectively. Twenty-six percent, 38%, and 29% of PJIs were treated with two-stage exchange, debridement and retention, or resection arthroplasty, respectively, and the 2-year survival rate free of treatment failure in each group was 77.7% (95% confidence interval, 42.8%-94.2%), 52.7% (95% confidence interval, 28.4%-75.9%), and 64.2% (95% confidence interval, 28.7%-88.9%). Methicillin-resistant Staphylococcus aureus (26.4% versus 7.1%) and anaerobes (11.7% versus 2.8%) were more common in polymicrobial PJIs. Polymicrobial PJIs occurred in patients with a soft tissue defect/dehiscence (23.5% versus 2.8%), drainage (79.4% versus 39.2%), or prior local irradiation (8.8% versus 0.71%). We found the following factors associated with polymicrobial prosthetic joint infections: the presence of a soft tissue defect/wound dehiscence (odds ratio, 5.9), drainage (odds ratio, 5.0), and age 65 years or older (odds ratio, 2.8). LEVEL OF EVIDENCE: Level III, prognostic study. See the Guidelines for Authors for a complete description of levels of evidence. PMID- 18421539 TI - Hematogenous septic ankle arthritis. AB - Ankle infection is a serious problem with limited published information on microbiology and associated morbidities. We describe the laboratory findings, microbiology, and occurrence of adjacent osteomyelitis in patients with hematogenous septic ankle arthritis. We retrospectively reviewed 30 patients with hematogenous septic arthritis of the ankle admitted over a 10-year period. Twenty two patients were male and eight female. The mean age was 46 years (range, 23-67 years). C-reactive protein and erythrocyte sedimentation rate were elevated in all patients, but the peripheral white blood cell count was elevated in only 47% of patients. Staphylococcus aureus (S. aureus) was the most common pathogen, isolated in 13 (54%) of the 24 patients with positive cultures; four of these isolates (four of 24; 17% of positive cultures) were oxacillin-resistant. Four (17%) of the 24 patients with positive cultures had a mycobacterial infection. We identified adjacent osteomyelitis in 30% of patients, which was considerably associated with the presence of patient comorbidities. S. aureus is the most common pathogen in septic ankle arthritis and empiric antibiotic therapy is recommended. Adjacent osteomyelitis may be present and a high index of suspicion is necessary in patients with comorbidities. LEVEL OF EVIDENCE: Level III, diagnostic study. See the Guidelines for Authors for a complete description of levels of evidence. PMID- 18421540 TI - Chitosan films: a potential local drug delivery system for antibiotics. AB - Local antibiotic delivery is an emerging area of study designed to provide alternative methods of treatment to clinicians for compromised wound sites where avascular zones can prevent the delivery of antibiotics to the infected tissue. Antibiotic-loaded bone cement is the gold standard for drug-eluting local delivery devices but is not ideal because it requires a removal surgery. Chitosan is a biocompatible, biodegradable polymer that has been used in several different drug delivery applications. We evaluated chitosan as a potential localized drug delivery device. We specifically determined if chitosan could elute antibiotics in an active form that would be efficacious in inhibiting S. aureus growth. Elution of amikacin was 24.67 +/- 2.35 microg/mL (85.68%) after 1 hour with a final cumulative release of 27.31 +/- 2.86 microg/mL (96.23%) after 72 hours. Elution of daptomycin was 10.17 +/- 3.83 microg/mL after 1 hour (31.61% release) and 28.72 +/- 6.80 microg/mL after 72 hours (88.55%). The data from the elution study suggested effective release of amikacin and daptomycin. The activity studies indicated the eluants inhibited the growth of S. aureus. Incorporating antibiotics in chitosan could provide alternative methods of treating musculoskeletal infections. PMID- 18421541 TI - Infection associated with hematoma formation after shoulder arthroplasty. AB - Hematoma formation requiring operative treatment after shoulder arthroplasty may be associated with higher patient morbidity. We therefore determined whether there was an association of hematoma formation requiring operative treatment with deep infection after shoulder arthroplasty. Between 1978 and 2006, we performed 4147 shoulder arthroplasties in 3643 patients. Of these, 12 shoulders (0.3%) underwent reoperation for hematoma formation. The mean time interval from arthroplasty to surgery for the hematoma was 7 days (range, 0.5-31 days). Among nine cases in which cultures were taken, six had positive cultures; the organisms included Propionibacterium acnes in three, Staphylococcus epidermidis in one, Streptococcus species in one, and Staphylococcus epidermidis with Peptostreptococcus in one. The minimum followup was 12 months (mean, 68 months; range, 12 to 294 months). Two of the 12 patients eventually underwent resection arthroplasty for deep infection. The Neer score was excellent in one, satisfactory in six, and unsatisfactory in five patients. The data suggest hematoma formation after shoulder arthroplasty is often accompanied by positive intraoperative cultures. The surgeon should be aware of the high rate of unsatisfactory results associated with this complication as well as the possibility of developing a deep infection requiring additional surgery. LEVEL OF EVIDENCE: Level IV, prognostic study. See Guidelines for Authors for a complete description of levels of evidence. PMID- 18421542 TI - Periprosthetic joint infection: the incidence, timing, and predisposing factors. AB - Periprosthetic joint infection is one of the most challenging complications of joint arthroplasty. We identified current risk factors of periprosthetic joint infection after modern joint arthroplasty, and determined the incidence and timing of periprosthetic joint infection. We reviewed prospectively collected data from our database on 9245 patients undergoing primary hip or knee arthroplasty between January 2001 and April 2006. Periprosthetic joint infections developed in 63 patients (0.7%). Sixty-five percent of periprosthetic joint infections developed within the first year of the index arthroplasty. The infecting organism was isolated in 57 of 63 cases (91%). The most common organisms identified were Staphylococcus aureus and Staphylococcus epidermidis. We identified the following independent predictors for periprosthetic joint infection: higher American Society of Anesthesiologists score, morbid obesity, bilateral arthroplasty, knee arthroplasty, allogenic transfusion, postoperative atrial fibrillation, myocardial infarction, urinary tract infection, and longer hospitalization. This study confirmed some previously implicated factors and identified new variables that predispose patients to periprosthetic joint infection. LEVEL OF EVIDENCE: Level II, prognostic study. PMID- 18421543 TI - Locally applied simvastatin promotes fracture healing in ovariectomized rat: a novel molecular mechanism. PMID- 18421544 TI - Rapid correction of low vitamin D status in nursing home residents. AB - This prospective study finds that ergocalciferol 50,000 IU three times weekly for four weeks effectively and safely corrects vitamin D inadequacy in nursing home residents. INTRODUCTION: Low vitamin D status is common among nursing home residents and contributes to bone loss, falls and fractures. The objective of this study was to evaluate the efficacy and safety of short course, high dose, oral vitamin D(2) (ergocalciferol) treatment. METHODS: This prospective study included 63 nursing home residents. The 25 with low vitamin D status (serum 25(OH)D < or = 25 ng/ml) received oral ergocalciferol 50,000 IU three times weekly for four weeks; the others received no change to their routine care. Serum total 25(OH)D, 25(OH)D(2), 25(OH)D(3), calcium, parathyroid hormone (PTH), bone turnover markers and neuro-cognitive assessments were obtained at baseline and four weeks. RESULTS: Mean total 25(OH)D concentration increased (p < 0.0001) from 17.3 to 63.8 ng/ml in the treated group and remained unchanged in the comparison group. Serum 25(OH)D(3) remained stable in the comparison group, but declined (p < 0.0001) with D(2) treatment from 15.4 to 9.1 ng/ml. Serum PTH trended down in the treatment group (p = 0.06). No treatment-induced improvement in ambulation, cognition or behavior was observed. No hypercalcemia or other adverse effects were observed with ergocalciferol treatment. CONCLUSION: Four weeks of oral vitamin D(2) supplementation effectively and safely normalizes serum 25(OH)D in nursing home residents. PMID- 18421545 TI - Lipoprotein subclasses and particle size determined by nuclear magnetic resonance spectroscopy in systemic lupus erythematosus. AB - Patients with systemic lupus erythematosus (SLE) have accelerated atherosclerosis, but the underlying mechanisms are unclear. The size and number of lipoprotein particles may be better predictors of atherosclerosis than conventional cholesterol measurements. We measured lipoprotein subclasses by nuclear magnetic resonance spectroscopy (NMR), coronary artery calcification by electron beam computed tomography, and insulin resistance by homeostasis model assessment in 105 patients with SLE and 77 control subjects. VLDL particles were larger (50.0+/-8.5 versus 47.7+/-8.5 nm, P=0.01) and concentrations of large high density lipoprotein (HDL) particles lower (10.1+/-5.3 versus 11.3+/-5.1 nmol/L, P=0.03) in patients with SLE than controls. In patients with SLE, small LDL concentration was associated with body mass index (rho=0.27), insulin resistance (rho=0.34), C-reactive protein (CRP; rho=0.30), and erythrocyte sedimentation rate (ESR; rho=0.20); all P<0.05. Large HDL concentration was inversely associated with insulin resistance (rho=-0.29), disease activity (rho=-0.23), and ESR (rho=-0.39); all P<0.05. VLDL concentrations correlated with CRP (rho=0.22), ESR (rho=0.24), disease damage (rho=0.20), and corticosteroid exposure (rho=0.29); all P<0.05. Neither the concentration of lipoprotein subclasses nor particle size was associated with coronary artery atherosclerosis. There were only minor differences in the NMR lipid profiles of patients with SLE and controls. Lipoprotein subclasses were associated with metabolic variables, inflammatory markers, and corticosteroid use but not with coronary artery atherosclerosis in SLE. PMID- 18421546 TI - Influence of gender and age of onset on the outcome in children with systemic lupus erythematosus. AB - The objective of our study was to determine the influence of gender and age of onset on the outcome in Saudi children with systemic lupus erythematosus (SLE). Medical records of children with SLE treated at King Faisal Specialist Hospital and Research Center were reviewed. Outcome measures included Systemic Lupus International Collaborating Clinics/American College of Rheumatology Damage Index score (SLICC/ACR), renal disease requiring dialysis, or transplant and death related to SLE. Patients were classified based on age at disease onset into early onset (<5 years) and late onset (>5 years). Data were analyzed, and comparison was made according to the gender and age groups. Eighty-nine patients (76 female and 13 male) were included. The median disease duration was 5 years. Twelve patients had early-onset disease. There was no difference in the mean age, age at diagnosis, disease duration, and follow-up between the different groups. Logistic regression analysis showed significant association of high SLICC/ACR score with early-onset disease and male gender, while renal disease requiring dialysis and renal transplant was associated significantly with male gender independently of age of disease onset. In contrast, death related to SLE was influenced by early onset disease. Male children and early onset disease of this cohort had poorer outcome. This finding indicates that gender and early-onset disease influence the long-term outcome of SLE in children. PMID- 18421547 TI - Regulation of the plasma membrane during exposure to low temperatures in suspension-cultured cells from a cryophyte (Chorispora bungeana). AB - As the outermost boundary of the cell, the plasma membrane plays an important role in determining the stress resistance of organisms. To test this concept in a cryophyte, we analyzed alterations of several components in plasma membranes isolated from suspension-cultured cells of Chorispora bungeana Fisch. & C.A. Mey in response to treatment at 0 and -4 degrees C for 192 h. When compared with the controls growing at 25 degrees C, both the membrane permeability and fluidity showed recovery after the initial impairment. Linolenic acid and membrane lipid unsaturation increased by about 0.8-fold following cold treatments, although the kinetics of the increase varied with the temperatures examined. During the treatments, the plasma membrane H(+)-ATPase (EC 3.6.1.3) activity increased by 78.06% at 0 degrees C and 100.47% at -4 degrees C. However, the plasma membrane NADH oxidase (EC 1.6.99.3) activity only decreased when exposed to a lower temperature (-4 degrees C), and remained at 63.93% after being treated for 192 h. After the treatments, the physical properties of the plasma membranes of suspension-cultured cells, especially the -4 degrees C treated cells, were similar to those in the wild plants. These findings indicate that the specific mechanism of cold resistance of C. bungeana is tightly linked with the rapid and flexible regulation of membrane lipids and membrane-associated enzymes, which ensure the structural and functional integrity of the plasma membrane that is essential for withstanding low temperature. PMID- 18421549 TI - Structural and immunocytochemical characterization of the adhesive tendril of Virginia creeper (Parthenocissus quinquefolia [L.] Planch.). AB - The tendrils of Virginia creeper (Parthenocissus quinquefolia) do not coil around their supports. Rather, they adhere to supporting objects by flattening against the support surface and secreting an adhesive compound which firmly glues the tendril to the support. In this study, microscopic and immunocytochemical techniques were utilized to determine the nature of this adhesive. Following touch stimulation, epidermal cells of the tendril elongate toward the support substrate, becoming papillate in morphology. Following contact with the support surface, an adhesive is produced at the base of the papillate cells. The adhesive appears as a highly heterogeneous, raftlike structure and consists of pectinaceous, rhamnogalacturonan (RG) I-reactive components surrounding a callosic core. In addition, more mobile components, composed of arabinogalactans and mucilaginous pectins, intercalate both the support and the tendril, penetrating the tendril to the proximal ends of the papillate cells. Following adherence to the support, the anticlinal walls of the papillate cells are devoid of RG I side-chain reactivity, indicating that extensive debranching of RG I molecules has taken place. Furthermore, a large amount of RG I backbone reactivity was observed in the contact area. These results may indicate that the debranched RG I molecules diffuse into and permeate the contact region, forming an integral part of the adhesive compound. These results indicate that Virginia creeper adheres to objects by a composite adhesive structure consisting of debranched RG I, callose, and other, less-well characterized mucilaginous pectins and that this structure subsequently becomes lignified and very weather-resistant upon the ultimate senescence of the tendril. PMID- 18421548 TI - An F-actin-depleted zone is present at the hyphal tip of invasive hyphae of Neurospora crassa. AB - The distribution of filamentous actin (F-actin) in invasive and noninvasive hyphae of the ascomycete Neurospora crassa was investigated. Eighty six percent of noninvasive hyphae had F-actin in the tip region compared to only 9% of invasive hyphae. The remaining 91% of the invasive hyphae had no obvious tip high concentration of F-actin staining; instead they had an F-actin-depleted zone in this region, although some F-actin, possibly associated with the Spitzenkorper, remained at the tip. The size of the F-actin-depleted zone in invasive hyphae increased with an increase in agar concentration. The membrane stain FM 4-64 reveals a slightly larger accumulation of vesicles at the tips of invasive hyphae relative to noninvasive hyphae, although this difference is unlikely to be sufficient to account for the exclusion of F-actin from the depleted zone. Antibodies raised against the actin filament-severing protein cofilin from both yeast and human cells localize to the tips of invasive hyphae. The human cofilin antibody shows a more random distribution in noninvasive hyphae locating primarily at the hyphal periphery but with some diffuse cytoplasmic staining. This antibody also identifies a single band at 21 kDa in immunoblots of whole hyphal fractions. These data suggest that a protein with epitopic similarity to cofilin may function in F-actin dynamics that underlie invasive growth. The F actin-depleted zone may play a role in the regulation of tip yielding to turgor pressure, thus increasing the protrusive force necessary for invasive growth. PMID- 18421550 TI - What generates flux of tubulin in kinetochore microtubules? AB - We discuss models for production of tubulin flux in kinetochore microtubules. Current models concentrate solely on microtubules and their associated motors and enzymes. For example, in some models the driving force for flux is enzymes at the poles and the kinetochores; in others the driving force is motor molecules that are associated with a stationary spindle matrix. We present a different viewpoint, that microtubules are propelled poleward by forces arising from the spindle matrix, that the forces on the microtubules "activate" polymerising and depolymerising enzymes at kinetochores and poles, that matrix forces utilise actin, myosin, and microtubule motors, and that the matrix itself may not necessarily be static. PMID- 18421551 TI - CDKA and CDKB kinases from Chlamydomonas reinhardtii are able to complement cdc28 temperature-sensitive mutants of Saccharomyces cerevisiae. AB - Cyclin-dependent kinases (CDK) play a key role in coordinating cell division in all eukaryotes. We investigated the capability of cyclin-dependent kinases CDKA and CDKB from the green alga Chlamydomonas reinhardtii to complement a Saccharomyces cerevisiae cdc28 temperature-sensitive mutant. The full-length coding regions of algal CDKA and CDKB cDNA were amplified by RT-PCR and cloned into the yeast expression vector pYES-DEST52, yielding pYD52-CDKA and pYD52-CDKB. The S. cerevisiae cdc28-1N strain transformed with these constructs exhibited growth at 36 degrees C in inducing (galactose) medium, but not in repressing (glucose) medium. Microscopic observation showed that the complemented cells had the irregular cylindrical shape typical for G2 phase-arrested cells when grown on glucose at 36 degrees C, but appeared as normal budded cells when grown on galactose at 36 degrees C. Sequence analysis and complementation tests proved that both CDKA and CDKB are functional CDC28/cdc2 homologs in C. reinhardtii. The complementation of the mitotic phenotype of the S. cerevisiae cdc28-1N mutant suggests a mitotic role for both of the kinases. PMID- 18421552 TI - Cellular expansion and gene expression in the developing grape (Vitis vinifera L.). AB - Expression profiles of genes involved in cell wall metabolism and water transport were compared with changes in grape (Vitis vinifera L.) berry growth, basic chemical composition, and the shape, size, and wall thickness of cells within tissues of the berry pericarp. Expression of cell wall-modifying and aquaporin genes in berry pericarp tissues generally followed a bimodal expression profile with high levels of expression coinciding with the two periods of rapid berry growth, stages I and III, and low levels of expression corresponding to the slow growth period, stage II. Cellular expansion was observed throughout all tissues during stage I, and only mesocarp cellular expansion was observed during stage III. Expansion of only exocarp cells was evident during transition between stages II and III. Cell wall-modifying and aquaporin gene expression profiles followed similar trends in exocarp and mesocarp tissues throughout berry development, with the exception of the up-regulation of pectin methylesterase, pectate lyase, two aquaporin genes (AQ1 and AQ2), and two expansin genes (EXP3 and EXPL) during stage II, which was delayed in the exocarp tissue compared with mesocarp tissue. Exocarp endo-(1-->3)-beta-glucanase and expansin-like gene expression was concurrent with increases in epidermal and hypodermal cell wall thickness. These results indicate a potential role of the grape berry skin in modulating grape berry growth. PMID- 18421553 TI - [Cardiac Imaging 2008]. PMID- 18421554 TI - [Usefulness of cardiovascular magnetic resonance imaging for the detection of coronary artery disease]. AB - Currently, the main focus of non-invasive cardiovascular magnetic resonance imaging for diagnosis of coronary artery disease is twofold: first, functional imaging with assessment of the hemodynamic consequences of obstructive coronary artery disease and second, anatomical imaging with direct visualization of the coronary arterial tree. This manuscript will provide an overview of the cardiovascular magnetic resonance imaging modules used to detect coronary artery disease and will update the reader on its current clinical indications with a special focus on functional versus anatomical imaging. PMID- 18421555 TI - Ablation of atrial fibrillation using CT image integration. AB - In our case report we describe a catheter ablation in a patient with symptomatic, drug-refractory atrial fibrillation using the new CT image integrating mapping technology. Using image integration an arrhythmogenic right inferior pulmonary vein was revealed which was not found during conventional 3-dimensional mapping. Thereby we want to demonstrate potential impacts on safety and effectiveness of ablation strategies using an image integration technology, especially in cases of difficult anatomic variations. PMID- 18421556 TI - Aortic valve endocarditis due to abiotrophia defectiva: a rare etiology. AB - Abiotrophia defectiva is a rare cause of infective endocarditis. Besides an association with often negative blood cultures and difficult treatment, high rates of relapse and higher mortality than endocarditis caused by other viridans streptococci are known features of this entity. We report on the surgical and medical management of the aortic valve endocarditis caused by Abiotrophia defectiva in a 19-year-old patient. To the best of our knowledge, this is the first case of a patient to have Down syndrome and Abiotrophia defectiva endocarditis requiring aortic valve replacement. PMID- 18421557 TI - [Application of CPAP improves oxygenation during normobaric and hypobaric hypoxia]. AB - This pilot project was performed to investigate the possible application of a portable CPAP helmet (continuous positive airway pressure) to treat high altitude diseases. Arterial oxygen saturation increased from 80.6 +/- 3.4 % to 90.6 +/- 3.8 % (p < 0.01, n = 14) when using the CPAP helmet (PEEP (positive endexpiratory pressure): 10-15 cm H2O) in normobaric hypoxia (FiO2 12.9 +/- 0.4 %, corresponding to about 4300 m). Furthermore arterial oxygen saturation was enhanced (88.1 +/- 1.9 % to 93.5 +/- 3.0 %; p < 0.01, n = 11) at real altitude (3150 m) after a 20 minute application of the CPAP helmet. There was a correlation between differences in PaCO2 and arterial oxygen saturation during CPAP (R = -0.97, p < 0.01, n = 6). Further studies are needed to determine the effectiveness, its mechanisms in context with altitude diseases, and the applicability of this treatment under extreme environmental conditions. PMID- 18421558 TI - [Characteristics of case management programs and their potential for patient empowerment]. AB - BACKGROUND AND AIM: Different types of case management programs have been increasingly developed for health care systems. This study, therefore, aimed to identify similarities and differences of case management programs, to estimate their shaping with special emphasis on patient empowerment, and to develop an instrument for their differentiation. METHODS: During an interdisciplinary seminar for students, eight different case management programs were selected and analyzed. RESULTS: Five characteristics of case management programs were identified: type of case, extent of functions, players involved, limits of availability and need for resources. The programs were categorized into information-, provider-, disease- or patient centred. With these characteristics the selected programs could be exactly described and their potential for patient empowerment could be assessed. CONCLUSIONS: This newly developed tool for describing case management programs allows for their sufficient differentiation. The potential for patient empowerment apparently is most prominent in patient centred programs. PMID- 18421559 TI - Early Austrian multicenter experience with palonosetron as antiemetic treatment for patients undergoing highly or moderately emetogenic chemotherapy. AB - BACKGROUND: Palonosetron is a new generation 5-HT3-receptor antagonist with a significantly prolonged half-life and a once-a-day administration compared to the conventional setrons. To evaluate the antiemetic efficacy of palonosetron in the daily hospital practice setting, a postmarketing study was carried out in Austria. METHODS: Palonosetron was administered at 0.25 mg on day 1 of each cycle to 135 cancer patients who received moderately or highly emetogenic chemotherapy either as an IV bolus or as a short-term infusion. Two thirds of these patients were females (n = 90), the majority had breast cancer (n = 38) and the majority received cisplatin, carboplatin, anthracyclines, 5-fluorouracil or cyclophosphamide. RESULTS: The complete antiemetic response rate was 68 % overall with 87 % efficacy on day 1 and 72 % efficacy on days 2 to 5. Higher antiemetic response was achieved in male patients, in patients being aged > or = 50 years, and in chemonaive patients. Twenty-four percent of patients needed rescue medication. Only 1.5 % of patients reported mild adverse events. CONCLUSIONS: Palonosetron resulted in high antiemetic efficacy in this study. Female gender and age < or = 50 years should be particularly considered when the antiemetic regimen is selected. PMID- 18421561 TI - [Curing aging?]. AB - The growing rate of high aged patients confronts medical care and nursing with diverse challenges which need interdisciplinary solutions. Experts in Medicine, Nursing Science, Palliative Care, Philosophy, Jurisprudence, Theology, Gerontology and Sociology joined the Conference "Curing of Aging?", Oct. 30, 2007 in Vienna and discussed central questions: The cooperation of Cure and Care and relevant research, principles concerning living wills and the relationship between society and people with dementia. PMID- 18421560 TI - [Cohort studies]. AB - This article about cohort studies is part of a methods series about study designs and their critical evaluation by the Ludwig Boltzmann Institute for Health Technology assessment. This article aims to describe the theoretical concept of cohort studies and their typical characteristics. Furthermore, it strives to highlight advantages and disadvantages of this study type and to make suggestions for the critical evaluation of the significance and validity of cohort studies. The article gives an account about characteristics due to the observational design and ways of acquiring control groups. Problems of blurring results by selection bias and confounding are also discussed. Cohort studies are applied in situations where the effects of environmental exposures are measured and rare side effects are identified but randomised controlled studies did not show significant results because of limitations. They are also used to assess the incidence of a disease or a condition. PMID- 18421562 TI - Bridging protein local structures and protein functions. AB - One of the major goals of molecular and evolutionary biology is to understand the functions of proteins by extracting functional information from protein sequences, structures and interactions. In this review, we summarize the repertoire of methods currently being applied and report recent progress in the field of in silico annotation of protein function based on the accumulation of vast amounts of sequence and structure data. In particular, we emphasize the newly developed structure-based methods, which are able to identify locally structural motifs and reveal their relationship with protein functions. These methods include computational tools to identify the structural motifs and reveal the strong relationship between these pre-computed local structures and protein functions. We also discuss remaining problems and possible directions for this exciting and challenging area. PMID- 18421563 TI - Solution structure of NPr, a bacterial signal-transducing protein that controls the phosphorylation state of the potassium transporter-regulating protein IIA Ntr. AB - A nitrogen-related signal transduction pathway, consisting of the three phosphotransfer proteins EI(Ntr), NPr, and IIA(Ntr), was discovered recently to regulate the uptake of K(+) in Escherichia coli. In particular, dephosphorylated IIA(Ntr) inhibits the activity of the K(+) transporter TrkA. Since the phosphorylation state of IIA(Ntr) is partially determined by its reversible phosphorylation by NPr, we have determined the three-dimensional structure of NPr by solution NMR spectroscopy. In total, we obtained 973 NOE-derived distance restraints, 112 chemical shift-derived backbone angle restraints, and 35 hydrogen bond restraints derived from temperature coefficients (wave). We propose that temperature wave is useful for identifying exposed beta-strands and assists in establishing protein folds based on chemical shifts. The deduced structure of NPr contains three alpha-helices and four beta-strands with the three helices all packed on the same face of the beta-sheet. The active site residue His16 of NPr for phosphoryl transfer was found to be neutral and in the N epsilon 2-H tautomeric state. There appears to be increased motion in the active site region of NPr compared to HPr, a homologous protein involved in the uptake and regulation of carbohydrate utilization. PMID- 18421564 TI - Cryptic species in the fern Ceratopteris thalictroides (Parkeriaceae). III. Referential diagnostic characters of three cryptic species. AB - Three cryptic species of Ceratopteris thalictroides, named the south type, the north type and the third type, were examined for their morphological characteristics, using sporophytes cultivated under common conditions. The discriminant analysis for leaf characters followed by one-way layout ANOVA or Kruskal-Wallis test for selected combinations of characters revealed that the following characters may be effective for identifying the three types: the relative lengths of stipe to blade and to pinna, the degree of dissection, the segment densities on rachis and pinna rachis, and the elongation degree of ultimate segments. The number of annulus cells on sporangia also proved to be a possible distinguishing character. As morphological data were obtained from a limited number of cultivated sporophytes, they are regarded as not definitive, but only referential diagnostic characters of the types and should be utilized not solely, but collectively, to avoid identification errors of the types. An identification trial using herbarium specimens proved these diagnostic characters to be useful to a considerable degree. PMID- 18421565 TI - Acaricidal effects of natural six-carbon and nine-carbon aldehydes on stored product mites. AB - The toxicities of three plant volatiles, (2E)-hexenal, (2E, 6Z)-nonadienal and (2E)-nonenal, intermediate products of the oxylipin biosynthesis pathway, were tested on three mites of importance for medical purposes and as pests. The aldehydes were diluted in hexane separately and incorporated into diets in ranges of 4-143 mg g(-1). The final density of mites in control and aldehyde-enriched diets was compared after 21 days. The aldehydes were toxic to the mites, whose final density showed an inverse correlation with aldehyde concentration. In addition to the effects of aldehyde concentration, the final density of mites was also influenced by the different aldehydes tested and the interaction among aldehyde concentration and chemical structure. In a functional combination of aldehydes and species, the doses calculated for growth inhibition and eradication of mites ranged from 4 to 35 mg g(-1) and from 36 to 314 mg g(-1), respectively. Due to the protective role displayed by natural six-carbon and nine-carbon aldehydes, these compounds are potential candidates for controlling stored product mites in stored food and feed products. PMID- 18421566 TI - Bone resorption increases tumour growth in a mouse model of osteosclerotic breast cancer metastasis. AB - Osteosclerotic metastases account for 20% of breast cancer metastases with the remainder osteolytic or mixed. In mouse models, osteolytic metastases are dependent on bone resorption for their growth. However, whether the growth of osteosclerotic bone metastases depends on osteoclast or osteoblast actions is uncertain. In this study, we investigate the effects of high and low bone resorption on tumour growth in a mouse model of osteosclerotic metastasis. We implanted human breast cancer, MCF-7, cells into the tibiae of mice. Low and high levels of bone resorption were induced by osteoprotegerin (OPG) treatment or calcium deficient diet respectively. We demonstrate that OPG treatment significantly reduces tumour area compared to vehicle (0.42 +/- 0.06 vs. 1.27 +/- 0.16 mm2, P < 0.01) in association with complete inhibition of osteoclast differentiation. In contrast, low calcium diet increases tumour area compared to normal diet (0.90 +/- 0.30 vs. 0.58 +/- 0.20 mm2, P < 0.05) in association with increased osteoclast numbers (84.44 +/- 5.18 vs. 71.11 +/- 3.56 per mm2 bone lesion area, P < 0.05). Osteoblast surfaces and new woven bone formation were similarly increased within the tumour boundaries in all treatment groups. Tumour growth in this model of osteosclerotic metastasis is dependent on ongoing bone resorption, as has been observed in osteolytic models. Bone resorption, rather than bone formation, apparently mediates this effect as osteoblast surfaces in the tumour mass were unchanged by treatments. Treatment of breast cancer patients through correction of calcium deficiency and/or with anti-resorptive agents such as OPG, may improve patient outcomes in the adjuvant as well as palliative settings. PMID- 18421567 TI - Structural characterization and functional properties of a novel lipomannan variant isolated from a Corynebacterium glutamicum pimB' mutant. AB - The genus Corynebacterium is part of the phylogenetic group nocardioform actinomycetes, which also includes the genus Mycobacterium. Members of this phylogenetic group have a characteristic cell envelope structure, which is dominated by complex lipids and amongst these, lipoglycans are of particular interest. The disruption of NCgl2106 in C. glutamicum resulted in a mutant devoid of monoacylated phosphatidyl-myo-inositol dimannoside (Ac(1)PIM(2)) resulting in the accumulation of Ac(1)PIM(1) and cessation of phosphatidyl-myo-inositol (PI) based lipomannan (Cg-LM, now also termed 'Cg-LM-A') and lipoarabinomannan (Cg LAM) biosynthesis. Interestingly, SDS-analysis of the lipoglycan fraction from the mutant revealed the synthesis of a single novel lipoglycan, now termed 'Cg-LM B'. Further chemical analyses established the lipoglycan possessed an alpha-D: glucopyranosyluronic acid-(1 --> 3)-glycerol (GlcAGroAc(2)) based anchor which was then further glycosylated by 8-22 mannose residues, with Man(12 20)GlcAGroAC(2) molecular species being the most abundant, to form a novel lipomannan structure (Cg-LM-B). The deletion of NCgl2106 in C. glutamicum has now provided a useful strain, in addition with a deletion mutant of NCgl0452 in C. glutamicum for the purification of Cg-LM-A and Cg-LM-B. Interestingly, both Cg-LM species induced a similar production of TNF-alpha by a human macrophage cell line suggesting that the phospho-myo-inositol residue of the PI-anchor does not play a key role in lipoglycan pro-inflammatory activity. PMID- 18421568 TI - In vivo analysis of chlorophyll a fluorescence induction. AB - Quantitative characteristics of photosynthetic electron transport were evaluated in vivo on the basis of the multi-exponential analysis of OJIP fluorescence transients induced by saturating actinic light. The OJIP fluorescence curve F(t), measured in Chlamydomonas reinhardtii cells, was transformed into the (1 - F(O)/F(t)) x (F(V)/F(M))(-1) transient, which is shown to relate to PS 2 closure. We assumed that kinetics of PS 2 closure during OJIP rise reflects time-separated processes related to the establishment of redox equilibrium at the PS 2 acceptor side (OJ), PQ pool (JI), and beyond Cyt b/f (IP). Three-exponential fitting was applied to (1 - F(O)/F(t)) x (F(V)/F(M))(-1) transient to obtain lifetimes and amplitudes of the OJ, JI, and IP components of PS 2 closure, which were used to calculate overall rates of reduction and re-oxidation of the PS 2 acceptor side, PQ pool, and intermediates beyond Cyt b/f complex. The results, obtained in the presence of inhibitors, oxidative reagents, and under different stress conditions prove the suggested model and characterize the introduced parameters as useful indicators of photosynthetic function. PMID- 18421569 TI - Lymphocutaneous sporotrichosis in an immunocompetent patient: a case report from extreme southern Italy. AB - Sporotrichosis is a mycotic infection rarely encountered in European countries, including Italy. We describe lymphocutaneous sporotrichosis in an immunocompetent 64-year-old male Italian mason. The patient had linear nodules along the right upper arm with severe lymphadenopathy in the same arm. Sporothrix schenckii was identified by phenotypic and molecular methods. Treatment with itraconazole and methylprednisolone followed by administration of itraconazole alone successfully cured the infection. To our knowledge, this is the first documented report of sporotrichosis from Calabria and Sicily, extreme southern Italy. PMID- 18421570 TI - Pseudocercospora cryptostegiae-madagascariensis sp. nov. on Cryptostegia madagascariensis, an exotic vine involved in major biological invasions in northeast Brazil. AB - Cryptostegia madagascariensis is a plant native from Madagascar, belonging to the Apocynaceae that is invading the native vegetation in Northeast Brazil and threatening the unique riverine formations dominated by the carnauba palm. Individuals of C. madagascariensis cultivated in the campus of the Universidade Federal de Vicosa, state of Minas Gerais, Brazil, showing leaf spot symptoms of unknown etiology were observed. Two fungal species were found associated to such leaf spots: Colletotrichum gloeosporioides and a new species of Pseudocercospora. The latter was named Pseudocercospora cryptostegiae-madagascariensis and described herein. The discovery of those two fungal pathogens on C. madagascariensis coincide with the recognition that the weedy vine that is involved in the infestations in the Northeast of Brazil is not Cryptostegia grandiflora as formerly reported. These mycological findings are of greater importance now since C. madagascariensis is likely to become a target for biological control together with its former status in Brazil of an ornamental plant of limited relevance. PMID- 18421571 TI - Competitiveness of a genetically engineered strain of Trichoderma virens. AB - The intraspecific competitiveness of a genetically engineered strain of Trichoderma virens was assessed relative to the non-transformed, progenitor strain and an isogenic, auxotrophic strain using a replacement series design. The transformed strain was less fit, but appeared more competitive than the wild type or the auxotroph in this assay. To validate this finding and to evaluate the ability of a strain to establish in an environment occupied by a competitor another experimental approach was developed. In various treatments the transformed strain was unaffected or only slightly inhibited by a competing strain. In contrast, the wild type and the auxotrophic strain were strongly inhibited by the presence of the transformed strain. These findings support the conclusion that this genetically engineered strain is more competitive than the wild-type strain and thus may be persistent in the environment. PMID- 18421572 TI - Different pathways in social support and quality of life between Korean American and Korean breast and gynecological cancer survivors. AB - PURPOSE: The purpose of this study was to investigate: (1) how social support influences quality of life (QOL) and (2) whether or not the pathways for predicting QOL outcomes differ between Korean American and Korean survivors of breast and gynecological cancer. METHOD: To identify multiple dimensions of health status and psychosocial outcomes, two standardized QOL and psychological distress measures were administered to 51 Korean Americans and 110 Koreans. Social support was measured by items assessing functional social support and social network structures. Hierarchical multiple regression and path analyses were employed to investigate the effects of social support on outcomes and to compare between Korean Americans and Koreans. RESULTS: The findings indicate that functional social support directly influences QOL and psychological distress, and social network structures are directly and/or indirectly associated with outcomes. Moreover, the two groups showed different pathways from social support to outcomes. CONCLUSION: This study provides practical research information by illuminating the relationships between social support components influencing QOL for Korean American and Korean survivors. This study may increase the utility and efficacy of the research and interventions to enhance cancer survivors' social participation and to diminish barriers to follow-up care, considering the different living conditions between the United States and Korea. PMID- 18421573 TI - Low level laser therapy (LLLT) decreases pulmonary microvascular leakage, neutrophil influx and IL-1beta levels in airway and lung from rat subjected to LPS-induced inflammation. AB - BACKGROUND AND OBJECTIVE: Low level laser therapy (LLLT) is a known anti inflammatory therapy. Herein we studied the effect of LLLT on lung permeability and the IL-1beta level in LPS-induced pulmonary inflammation. STUDY DESIGN/METHODOLOGY: Rats were divided into 12 groups (n = 7 for each group). Lung permeability was measured by quantifying extravasated albumin concentration in lung homogenate, inflammatory cells influx was determined by myeloperoxidase activity, IL-1beta in BAL was determined by ELISA and IL-1beta mRNA expression in trachea was evaluated by RT-PCR. The rats were irradiated on the skin over the upper bronchus at the site of tracheotomy after LPS. RESULTS: LLLT attenuated lung permeability. In addition, there was reduced neutrophil influx, myeloperoxidase activity and both IL-1beta in BAL and IL-1beta mRNA expression in trachea obtained from animals subjected to LPS-induced inflammation. CONCLUSION: LLLT reduced the lung permeability by a mechanism in which the IL-1beta seems to have an important role. PMID- 18421574 TI - 3D and 4D echo--applications in EP laboratory procedures. AB - INTRODUCTION: 3D echocardiography allows imaging and analysis of cardiovascular structures as they move in time and space, thus creating possibility for creation of 4D datasets (3D + time). Intracardiac echocardiography (ICE) further broadens the spectrum of echocardiographic techniques by allowing detailed imaging of intracardiac anatomy with 3D reconstructions. The paper reviews the current status of development of 3D and 4D echocardiography in electrophysiology. In ablation area, 3D echocardiography can enhance the performance of catheter ablation for complex arrhythmias such as atrial fibrillation. Currently, several strategies to obtain 3D reconstructions from ICE are available. One involves combination with electroanatomical mapping system; others create reconstruction from standard phased-array or single-element ICE catheter using special rotational or pull-back devices. Secondly, 3D echocardiography may be used for precise assessment of cardiac dyssynchrony before cardiac resynchronization therapy. Its reliable detection is expected to minimize number of non-responders to this treatment and optimize left ventricular lead positioning to get maximum hemodynamic benefit. CONCLUSION: The main potential benefit of 3D and 4D echocardiography in electrophysiology lie in real-time guidance of complex ablation procedures and precise assessment of cardiac dyssynchrony. PMID- 18421575 TI - Collective familial decision-making in times of trouble: intergenerational solidarity in Ghana. AB - In post-colonial populations the retention of the nuclear family concept of collectivism as the basis for decision-making is associated with familial and cultural survival. Collectivism within familial contexts provides intergenerational access to experience, information, resources, and sound decision making during times of conflict or dealing with the consequences of poverty. In contemporary Ghana inter-ethnic wars and conflict have marginalized minority groups, causing substantial internal displacement of people. Poverty and cultural issues have caused a persistently high level of child malnutrition and mortality in all areas of Ghana, including urban centers where appropriate food is in abundance. In each of these circumstances the senior women, particularly the grandmothers, have been found to be essential for child survival, protecting the family from dissolution, and ensuring intergenerational cultural transmission. This paper discusses the place of collective decision-making processes of older African women in two studies in Ghana that assessed the: (1) indigenous methods used for understanding and arresting violent ethnic conflict by internally displaced women who survived the 1994/95 Guinea Fowl War; and (2) parenting and grandparenting strategies from families who insured the survival of children suffering from kwashiorkor, a severe form of childhood malnutrition. In this paper, findings from qualitative interview data will include narratives and illustrative cases. PMID- 18421576 TI - Impact on survival of early detection of isolated breast recurrences after the primary treatment for breast cancer: a meta-analysis. AB - PURPOSE: The purpose was to establish the impact on survival of early detection of a local recurrence of breast cancer as compared to late detection. DESIGN: A meta-analysis was carried out using Cochrane review manager software (RevMan version 4.2). Studies were included if women were treated for primary breast cancer without evidence of distant metastasis at primary diagnosis and if these concerned routine follow-up strategies focusing on the early detection of curable recurrences. Data regarding the risk for death were derived from each study. Multi level models were used to study heterogeneity by using MLWin. RESULTS: Thirteen studies concerning 2,263 patients were included. Early detection of breast cancer recurrences during follow-up gave a significantly better survival as compared to late detected recurrences (HR: 1.68 (95% CI: 1.48-1.91)). Survival was better when the recurrence was found by mammography instead of physical examination or in patients without symptoms as compared to those with symptoms (HR: 2.44 (95% CI: 1.78-3.35); HR: 1.56 (95% CI: 1.36-1.79), respectively). If all breast cancer recurrences would be detected earlier, that 5-8 deaths (i.e. an absolute reduction in mortality of 17-28%) would be avoided by performing routine follow-up during a 10 year-period for 1,000 breast cancer patients. CONCLUSION: These data support the hypothesis that detection of isolated loco-regional or contra-lateral breast cancer recurrences in patients without symptoms has beneficial impact on survival of breast cancer patients when compared to late symptomatic detection. PMID- 18421577 TI - Therapeutic metformin/AMPK activation promotes the angiogenic phenotype in the ERalpha negative MDA-MB-435 breast cancer model. PMID- 18421578 TI - Death receptor-4 (DR4) expression is regulated by transcription factor NF-kappaB in response to etoposide treatment. AB - Tumour necrosis factor related apoptosis inducing ligand (TRAIL) binds to death receptor 4 (DR4) activating the apoptotic signalling pathway. DNA damaging agents (genotoxins) such as etoposide increase DR4 expression and when combined with TRAIL induce a synergistic apoptotic response. The mechanism for up-regulation of DR4 expression following genotoxin treatment is not well understood. Herein, we determined that transcription factor NF-kappaB plays a role in genotoxin induced DR4 expression. Increased expression of DR4 following etoposide treatment is blocked by inhibition of the NF-kappaB pathway. Moreover, expression of the p65 subunit of NF-kappaB is sufficient to increase DR4 protein levels. Indeed, knockdown of p65 by RNA interference blocked etoposide up-regulation of DR4. We further identified a functional NF-kappaB binding site located in the DR4 promoter. Mutation of this site abrogates the induction of luciferase activity after p65 over-expression. Furthermore, electromobility shift assays and chromatin immunoprecipitaton suggest that NF-kappaB binds to this site upon etoposide treatment. MEK kinase 1 (MEKK1) is a serine threonine kinase that is activated following etoposide treatment and activates NF-kappaB. Expression of the kinase inactive MEKK1 (MEKK1-KM) abrogates the up-regulation of DR4 after etoposide treatment. Taken together, NF-kappaB plays a role in up-regulation of DR4 following etoposide treatment. PMID- 18421581 TI - Transcranial surgery for pituitary adenomas. AB - Following a century of evolution and refinements in standard surgical techniques, the vast majority of operations for pituitary adenoma to date are performed utilizing transsphenoidal approaches. From current large series one obtains the impression that certainly less than 10% of these tumors require craniotomies. However, still several tumors, which's volume is mainly localized outside of the sella require transcranial approaches, of which the pterional and subfrontal routes are the most widely used. The goal of surgical treatment is rapid eradication of the tumor mass, decompression of visual pathways and elimination of hormonal oversecretion whilst preserving the normal gland and avoiding potential surgical complications. Even with microsurgical techniques and standardized approaches, there is still some mortality associated with transcranial approaches and morbidity is undoubtly higher than with transsphenoidal operations. However, a selection bias must be considered, which shifts tumors with a larger size, less favourable prognosis and higher complication rate into the transcranial series. Moreover, with extended transsphenoidal approaches, lesions have become accessible for transsphenoidal surgery, which previously have been considered as contraindications. In this article current indications and limitations for transcranial surgery of pituitary adenomas, the preoperative workup, surgical techniques, results, and complications are briefly reviewed. PMID- 18421580 TI - Androgen deficiency: effects on body composition. AB - Published data on the effects of androgen deficiency and testosterone administration on body composition in men and women are reviewed. In experimental paradigms, androgen deprivation decreases lean body mass and increases fat mass in men, and physiologic replacement reverses these abnormalities. The anabolic effects of testosterone administration on muscle in men are well-established, and current understanding of the underlying mechanisms are discussed. Randomized, placebo-controlled studies have been performed to investigate the effects of testosterone administration on body composition in a number of male hypogonadal states, including HIV-associated weight loss, supraphysiologic glucocorticoid administration, aging and obesity, with variable outcomes, and the results are reviewed. There are few data investigating the effects of hypoandrogenemia or androgen replacement on body composition in women, in whom endogenous testosterone levels are a fraction of those in men. A recent randomized, placebo controlled study of physiologic testosterone replacement therapy in women with profound hypoandrogenemia due to hypopituitarism demonstrated an increase in skeletal muscle mass but no change in body fat. Further research is needed to establish the effects of endogenous androgens on the regulation of body composition in women. PMID- 18421582 TI - Production of cyclodextrins by CGTase from Bacillus clausii using different starches as substrates. AB - Cyclodextrins (CDs) are cyclic oligasaccharides composed by D-glucose monomers joined by alpha-1,4-D glicosidic linkages. The main types of CDs are alpha-, beta and gamma-CDs consisting of cycles of six, seven, and eight glucose monomers, respectively. Their ability to form inclusion complexes is the most important characteristic, allowing their wide industrial application. The physical property of the CD-complexed compound can be altered to improve stability, volatility, solubility, or bio-availability. The cyclomaltodextrin glucanotransferase (CGTase, EC 2.4.1.19) is an enzyme capable of converting starch into CD molecules. In this work, the CGTase produced by Bacillus clausii strain E16 was used to produce CD from maltodextrin and different starches (commercial soluble starch, corn, cassava, sweet potato, and waxy corn starches) as substrates. It was observed that the substrate sources influence the kind of CD obtained and that this CGTase displays a beta-CGTase action, presenting a better conversion of soluble starch at 1.0%, of which 80% was converted in CDs. The ratio of total CD produced was 0:0.89:0.11 for alpha/beta/gamma. It was also observed that root and tuber starches were more accessible to CGTase action than seed starch under the studied conditions. PMID- 18421579 TI - HTLV-1 and apoptosis: role in cellular transformation and recent advances in therapeutic approaches. AB - A universal cellular defense mechanism against viral invasion is the elimination of infected cells through apoptotic cell death. To counteract host defenses many viruses have evolved complex apoptosis evasion strategies. The oncogenic human retrovirus HTLV-1 is the etiological agent of adult-T-cell leukemia/lymphoma (ATLL) and the neurodegenerative disease known as HTLV-associated myelopathy/tropical spastic paraparesis (HAM/TSP). The poor prognosis in HTLV-1 induced ATLL is linked to the resistance of neoplastic T cells against conventional therapies and the immuno-compromised state of patients. Nevertheless, several studies have shown that the apoptotic pathway is largely intact and can be reactivated in ATLL tumor cells to induce specific killing. A better understanding of the molecular mechanisms employed by HTLV-1 to counteract cellular death pathways remains an important challenge for future therapies and the treatment of HTLV-1-associated diseases. PMID- 18421583 TI - Effects of pH and temperature on recombinant manganese peroxidase production and stability. AB - The enzyme manganese peroxidase (MnP) is produced by numerous white-rot fungi to overcome biomass recalcitrance caused by lignin. MnP acts directly on lignin and increases access of the woody structure to synergistic wood-degrading enzymes such as cellulases and xylanases. Recombinant MnP (rMnP) can be produced in the yeast Pichia pastoris alphaMnP1-1 in fed-batch fermentations. The effects of pH and temperature on recombinant manganese peroxidase (rMnP) production by P. pastoris alphaMnP1-1 were investigated in shake flask and fed-batch fermentations. The optimum pH and temperature for a standardized fed-batch fermentation process for rMnP production in P. pastoris alphaMnP1-1 were determined to be pH 6 and 30 degrees C, respectively. P. pastoris alphaMnP1-1 constitutively expresses the manganese peroxidase (mnp1) complementary DNA from Phanerochaete chrysosporium, and the rMnP has similar kinetic characteristics and pH activity and stability ranges as the wild-type MnP (wtMnP). Cultivation of P. chrysosporium mycelia in stationary flasks for production of heme peroxidases is commonly conducted at low pH (pH 4.2). However, shake flask and fed-batch fermentation experiments with P. pastoris alphaMnP1-1 demonstrated that rMnP production is highest at pH 6, with rMnP concentrations in the medium declining rapidly at pH less than 5.5, although cell growth rates were similar from pH 4-7. Investigations of the cause of low rMnP production at low pH were consistent with the hypothesis that intracellular proteases are released from dead and lysed yeast cells during the fermentation that are active against rMnP at pH less than 5.5. PMID- 18421584 TI - Xylanase production by Bacillus circulans D1 using maltose as carbon source. AB - Bacillus circulans D1 is a good producer of extracellular thermostable xylanase. Xylanase production in different carbon sources was evaluated and the enzyme synthesis was induced by various carbon sources. It was found that D-maltose is the best inducer of the enzyme synthesis (7.05 U/mg dry biomass at 48 h), while D glucose and D-arabinose lead to the production of basal levels of xylanase. The crude enzyme solution is free of cellulases, even when the microorganism was cultivated in a medium with D-cellobiose. When oat spelt xylan was supplemented with D-glucose, the repressive effect of this sugar on xylanase production was observed at 24 h, only when used at 5.0 g/L, leading to a reduction of 60% on the enzyme production. On the other hand, when the xylan medium was supplemented with D-xylose (3.0 or 5.0 g/L), this effect was more evident (80 and 90% of reduction on the enzyme production, respectively). Unlike that observed in the xylan medium, glucose repressed xylanase production in the maltose medium, leading to a reduction of 55% on the enzyme production at 24 h of cultivation. Xylose, at 1.0 g/L, induced xylanase production on the maltose medium. On this medium, the repressive effect of xylose, at 3.0 or 5.0 g/L, was less expressive when compared to its effect on the xylan medium. PMID- 18421585 TI - Immobilization of fungal beta-glucosidase on silica gel and kaolin carriers. AB - Beta-glucosidase is a key enzyme in the hydrolysis of cellulose for producing feedstock glucose for various industrial processes. Reuse of enzyme through immobilization can significantly improve the economic characteristics of the process. Immobilization of the fungal beta-glucosidase by covalent binding and physical adsorption on silica gel and kaolin was conducted for consequent application of these procedures in large-scale industrial processes. Different immobilization parameters (incubation time, ionic strength, pH, enzyme/support ratio, glutaric aldehyde concentration, etc.) were evaluated for their effect on the thermal stability of the immobilized enzyme. It was shown that the immobilized enzyme activity is stable at 50 degrees C over 8 days. It has also been shown that in the case of immobilization on kaolin, approximately 95% of the initial enzyme was immobilized onto support, and loss of activity was not observed. However, covalent binding of the enzyme to silica gel brings significant loss of enzyme activity, and only 35% of activity was preserved. In the case of physical adsorption on kaolin, gradual desorption of enzyme takes place. To prevent this process, we have carried out chemical modification of the protein. As a result, after repeated washings, enzyme desorption from kaolin has been reduced from 75 to 20-25% loss. PMID- 18421586 TI - Immobilization of Yarrowia lipolytica lipase--a comparison of stability of physical adsorption and covalent attachment techniques. AB - Lipase immobilization offers unique advantages in terms of better process control, enhanced stability, predictable decay rates and improved economics. This work evaluated the immobilization of a highly active Yarrowia lipolytica lipase (YLL) by physical adsorption and covalent attachment. The enzyme was adsorbed on octyl-agarose and octadecyl-sepabeads supports by hydrophobic adsorption at low ionic strength and on MANAE-agarose support by ionic adsorption. CNBr-agarose was used as support for the covalent attachment immobilization. Immobilization yields of 71, 90 and 97% were obtained when Y. lipolytica lipase was immobilized into octyl-agarose, octadecyl-sepabeads and MANAE-agarose, respectively. However, the activity retention was lower (34% for octyl-agarose, 50% for octadecyl-sepabeads and 61% for MANAE-agarose), indicating that the immobilized lipase lost activity during immobilization procedures. Furthermore, immobilization by covalent attachment led to complete enzyme inactivation. Thermal deactivation was studied at a temperature range from 25 to 45 degrees C and pH varying from 5.0 to 9.0 and revealed that the hydrophobic adsorption on octadecyl-sepabeads produced an appreciable stabilization of the biocatalyst. The octadecyl-sepabeads biocatalyst was almost tenfold more stable than free lipase, and its thermal deactivation profile was also modified. On the other hand, the Y. lipolytica lipase immobilized on octyl-agarose and MANAE-agarose supports presented low stability, even less than the free enzyme. PMID- 18421587 TI - Heterologous expression of Aspergillus niger beta-D-xylosidase (XlnD): characterization on lignocellulosic substrates. AB - The gene encoding a glycosyl hydrolase family 3 xylan 1,4-beta-xylosidase, xlnD, was successfully cloned from Aspergillus niger strain ATCC 10864. The recombinant product was expressed in Aspergillus awamori, purified by column chromatography, and verified by matrix-assisted laser desorption ionization, tandem time of flight (MALDI-TOF/TOF) mass spectroscopy of tryptic digests. The T (max) was determined using differential scanning microcalorimetry (DSC) to be 78.2 degrees C; the K (m) and k (cat) were found to be 255 microM and 13.7 s(-1), respectively, using pNP-beta-D-xylopyranoside as substrate. End-product inhibition by D-xylose was also verified and shown to be competitive; the K (i) for this inhibition was estimated to be 3.3 mM. XlnD was shown to efficiently hydrolyze small xylo-oligomers to monomeric xylose, making it a critical hydrolytic activity in cases where xylose is to be recovered from biomass conversion processes. In addition, the presence of the XlnD was shown to synergistically enhance the ability of an endoxylanase, XynA from Thermomyces lanuginosus, to convert xylan present in selected pretreated lignocellulosic substrates. Furthermore, the addition of the XynA/XlnD complex was effective in enhancing the ability of a simplified cellulase complex to convert glucan present in the substrates. PMID- 18421588 TI - Cloning, expression and characterization of a glycoside hydrolase family 39 xylosidase from Bacillus halodurans C-125. AB - The gene encoding a glycoside hydrolase family 39 xylosidase (BH1068) from the alkaliphile Bacillus halodurans strain C-125 was cloned with a C-terminal His tag, and the recombinant gene product termed BH1068(His)(6) was expressed in Escherichia coli. Of the artificial substrates tested, BH1068(His)(6) hydrolyzed nitrophenyl derivatives of beta-D-xylopyranose, alpha-L-arabinofuranose, and alpha-L-arabinopyranose. Deviation from Michaelis-Menten kinetics at higher substrate concentrations indicative of transglycosylation was observed, and k (cat) and K (m) values were measured at both low and high substrate concentrations to illuminate the relative propensities to proceed along this alternate reaction pathway. The pH maximum was 6.5, and under the conditions tested, maximal activity was at 47 degrees C, and thermal instability occurred above 45 degrees C. BH1068(His)(6) was inactive on arabinan, hydrolyzed xylooligosaccharides, and released only xylose from oat, wheat, rye, beech, and birch arabinoxylan, and thus, can be classified as a xylosidase with respect to natural substrate specificity. The enzyme was not inhibited by up to 200 mM xylose. The oligomerization state was tetrameric under the size-exclusion chromatography conditions employed. PMID- 18421589 TI - Heterologous expression of two ferulic acid esterases from Penicillium funiculosum. AB - Two recombinant ferulic acid esterases from Penicillium funiculosum produced in Aspergillus awamori were evaluated for their ability to improve the digestibility of pretreated corn stover. The genes, faeA and faeB, were cloned from P. funiculosum and expressed in A. awamori using their native signal sequences. Both enzymes contain a catalytic domain connected to a family 1 carbohydrate-binding module by a threonine-rich linker peptide. Interestingly, the carbohydrate binding-module is N-terminal in FaeA and C-terminal in FaeB. The enzymes were purified to homogeneity using column chromatography, and their thermal stability was characterized by differential scanning microcalorimetry. We evaluated both enzymes for their potential to enhance the cellulolytic activity of purified Trichoderma reesei Cel7A on pretreated corn stover. PMID- 18421590 TI - Evaluation of a Hypocrea jecorina enzyme preparation for hydrolysis of Tifton 85 bermudagrass. AB - Tifton 85 bermudagrass, developed at the ARS-USDA in Tifton, GA, is grown on over ten million acres in the USA for hay and forage. Of the bermudagrass cultivars, Tifton 85 exhibits improved digestibility because the ratio of ether- to ester linked phenolic acids has been lowered using traditional plant breeding techniques. A previously developed pressurized batch hot water (PBHW) method was used to treat Tifton 85 bermudagrass for enzymatic hydrolysis. Native grass (untreated) and PBHW-pretreated material were compared as substrates for fungal cultivation to produce enzymes. Cellulase activity, measured via the filter paper assay, was higher for fungi cultivated on PBHW-pretreated grass, whereas the other nine enzyme assays produced higher activities for the untreated grass. Ferulic acid and vanillin levels increased significantly for the enzyme preparations produced using PBHW-pretreated grass and the release of these phenolic compounds may have contributed to the observed reduction in enzyme activities. Culture supernatant from Tifton 85 bermudagrass-grown fungi were combined with two commercial enzyme preparations and the enzyme activity profiles are reported. The amount of reducing sugar liberated by the enzyme mixture from Hypocrea jecorina (after 192 h incubation with untreated bermudagrass) individually or in combination with feruloyl esterase was 72.1 and 84.8%, respectively, of the commercial cellulase preparation analyzed under the same conditions. PMID- 18421591 TI - A novel technique that enables efficient conduct of simultaneous isomerization and fermentation (SIF) of xylose. AB - Of the sugars recovered from lignocellulose, D-glucose can be readily converted into ethanol by baker's or brewer's yeast (Saccharomyces cerevisiae). However, xylose that is obtained by the hydrolysis of the hemicellulosic portion is not fermentable by the same species of yeasts. Xylose fermentation by native yeasts can be achieved via isomerization of xylose to its ketose isomer, xylulose. Isomerization with exogenous xylose isomerase (XI) occurs optimally at a pH of 7 8, whereas subsequent fermentation of xylulose to ethanol occurs at a pH of 4-5. We present a novel scheme for efficient isomerization of xylose to xylulose at conditions suitable for the fermentation by using an immobilized enzyme system capable of sustaining two different pH microenvironments in a single vessel. The proof-of-concept of the two-enzyme pellet is presented, showing conversion of xylose to xylulose even when the immobilized enzyme pellets are suspended in a bulk solution whose pH is sub-optimal for XI activity. The co-immobilized enzyme pellets may prove extremely valuable in effectively conducting "simultaneous isomerization and fermentation" (SIF) of xylose. To help further shift the equilibrium in favor of xylulose formation, sodium tetraborate (borax) was added to the isomerization solution. Binding of tetrahydroxyborate ions to xylulose effectively reduces the concentration of xylulose and leads to increased xylose isomerization. The formation of tetrahydroxyborate ions and the enhancement in xylulose production resulting from the complexation was studied at two different bulk pH values. The addition of 0.05 M borax to the isomerization solution containing our co-immobilized enzyme pellets resulted in xylose to xylulose conversion as high as 86% under pH conditions that are suboptimal for XI activity. These initial findings, which can be optimized for industrial conditions, have significant potential for increasing the yield of ethanol from xylose in an SIF approach. PMID- 18421592 TI - The effects of wheat bran composition on the production of biomass-hydrolyzing enzymes by Penicillium decumbens. AB - The effects of the starch, protein, and soluble oligosaccharides contents in wheat bran on the extracellular biomass-hydrolyzing enzymes activities released by Penicillium decumbens mycelia grown in batch fermentations have been examined. The results showed increased starch content correlated directly with an increase in released amylase activity but inversely with the levels of secreted cellulase and xylanase. High amounts of protein in wheat bran also reduced the activities of cellulase, xylanase and protease in the culture medium. The effects of the soluble and insoluble components of wheat bran and cello-oligosaccharides supplements on production of extracellular cellulase and xylanase were compared. The soluble cello-oligosaccharides compositions in wheat bran were proved to be one of the most significant factors for cellulase production. According to the results of this research, determining and regulating the composition of wheat bran used as a fermentation supplement may allow for improved induction of cellulase and xylanase production. PMID- 18421594 TI - Beta-D-xylosidase from Selenomonas ruminantium: catalyzed reactions with natural and artificial substrates. AB - Catalytically efficient beta-D-xylosidase from Selenomonas ruminantium (SXA) exhibits pK (a)s 5 and 7 (assigned to catalytic base, D14, and catalytic acid, E186) for k (cat)/K (m) with substrates 1,4-beta-D-xylobiose (X2) and 1,4-beta-D xylotriose (X3). Catalytically inactive, dianionic SXA (D14(-)E186(-)) has threefold lower affinity than catalytically active, monoanionic SXA (D14( )E186(H)) for X2 and X3, whereas D14(-)E186(-) has twofold higher affinity than D14(-)E186(H) for 4-nitrophenyl-beta-D-xylopyranoside (4NPX), and D14(-)E186(-) has no affinity for 4-nitrophenyl-alpha-L-arabinofuranoside. Anomeric isomers, alpha-D-xylose and beta-D-xylose, have similar affinity for SXA. 4-Nitrophenol competitively inhibits SXA-catalyzed hydrolysis of 4NPX. SXA steady-state kinetic parameters account for complete progress curves of SXA-catalyzed hydrolysis reactions. PMID- 18421593 TI - Integrated biosensor systems for ethanol analysis. AB - Different integrated systems with a bi-enzymatic biosensor, working with two different methods for ethanol detection--flow injection analysis (FIA) or sequential injection analysis (SIA)--were developed and applied for ethanol extracted from gasohol mixtures, as well as for samples of alcoholic beverages and fermentation medium. A detection range of 0.05-1.5 g ethanol/l, with a correlation coefficient of 0.9909, has been reached when using FIA system, working with only one microreactor packed with immobilized alcohol oxidase and injecting free horseradish peroxidase. When using both enzymes, immobilized separately in two microreactors, the detection ranges obtained varied from 0.001 to 0.066 g ethanol/l, without on-line dilution to 0.010-0.047 g ethanol/l when a 1:7,000 dilution ratio was employed, reaching correlation coefficients of 0.9897 and 0.9992, respectively. For the integrated biosensor SIA system with the stop flow technique, the linear range was 0.005-0.04 g/l, with a correlation coefficient of 0.9922. PMID- 18421595 TI - Hydrolysis of ammonia-pretreated sugar cane bagasse with cellulase, beta glucosidase, and hemicellulase preparations. AB - Sugar cane bagasse consists of hemicellulose (24%) and cellulose (38%), and bioconversion of both fractions to ethanol should be considered for a viable process. We have evaluated the hydrolysis of pretreated bagasse with combinations of cellulase, beta-glucosidase, and hemicellulase. Ground bagasse was pretreated either by the AFEX process (2NH(3): 1 biomass, 100 degrees C, 30 min) or with NH(4)OH (0.5 g NH(4)OH of a 28% [v/v] per gram dry biomass; 160 degrees C, 60 min), and composition analysis showed that the glucan and xylan fractions remained largely intact. The enzyme activities of four commercial xylanase preparations and supernatants of four laboratory-grown fungi were determined and evaluated for their ability to boost xylan hydrolysis when added to cellulase and beta-glucosidase (10 filter paper units [FPU]: 20 cellobiase units [CBU]/g glucan). At 1% glucan loading, the commercial enzyme preparations (added at 10% or 50% levels of total protein in the enzyme preparations) boosted xylan and glucan hydrolysis in both pretreated bagasse samples. Xylanase addition at 10% protein level also improved hydrolysis of xylan and glucan fractions up to 10% glucan loading (28% solids loading). Significant xylanase activity in enzyme cocktails appears to be required for improving hydrolysis of both glucan and xylan fractions of ammonia pretreated sugar cane bagasse. PMID- 18421596 TI - Monoglycerides and diglycerides synthesis in a solvent-free system by lipase catalyzed glycerolysis. AB - Five lipases were screened (Thermomyces lanuginosus free and immobilized forms, Candida antarctica B, Candida rugosa, Aspergillus niger, and Rhizomucor miehei) to study their ability to produce monoglycerides (MG) and diglycerides (DG) through enzymatic glycerolysis of soybean oil. Lipase from C. antarctica was further studied to verify the enzyme load (wt% of oil mass), the molar ratio glycerol/oil, and the water content (wt% of glycerol) on the glycerolysis reaction. The best DG and MG productions were in the range 45-48% and 28-30% (w/w, based on the total oil), respectively. Using immobilized lipases, the amount of free fatty acids (FFA) produced was about 5%. However, the amount of FFA produced when using free lipases, with 3.5% extra water in the system, is equivalent to the MG yield, about 23%. The extra water content provides a competition between hydrolysis and glycerolysis reactions, increasing the FFA production. PMID- 18421597 TI - Immobilization of Candida antarctica lipase B by adsorption to green coconut fiber. AB - An agroindustrial residue, green coconut fiber, was evaluated as support for immobilization of Candida antarctica type B (CALB) lipase by physical adsorption. The influence of several parameters, such as contact time, amount of enzyme offered to immobilization, and pH of lipase solution was analyzed to select a suitable immobilization protocol. Kinetic constants of soluble and immobilized lipases were assayed. Thermal and operational stability of the immobilized enzyme, obtained after 2 h of contact between coconut fiber and enzyme solution, containing 40 U/ml in 25 mM sodium phosphate buffer pH 7, were determined. CALB immobilization by adsorption on coconut fiber promoted an increase in thermal stability at 50 and 60 degrees C, as half-lives (t (1/2)) of the immobilized enzyme were, respectively, 2- and 92-fold higher than the ones for soluble enzyme. Furthermore, operational stabilities of methyl butyrate hydrolysis and butyl butyrate synthesis were evaluated. After the third cycle of methyl butyrate hydrolysis, it retained less than 50% of the initial activity, while Novozyme 435 retained more than 70% after the tenth cycle. However, in the synthesis of butyl butyrate, CALB immobilized on coconut fiber showed a good operational stability when compared to Novozyme 435, retaining 80% of its initial activity after the sixth cycle of reaction. PMID- 18421598 TI - Methods and supports for immobilization and stabilization of cyclomaltodextrin glucanotransferase from Thermoanaerobacter. AB - Thermoanaerobacter cyclomaltodextrin glucanotransferase (CGTase) was immobilized using different supports and immobilization methods to study the effect on activity recovery. The enzyme covalently attached into glyoxyl-silica showed low activity recovery of 1.5%. The hydrophobic adsorption of the enzyme on Octadecyl Sepabeads yielded also low activity recovery, 3.83%, and the enzyme could easily leak from the support at low ionic strength, although the immobilization yield was satisfactory, approximately 76%. The CGTase encapsulated in a sol-gel matrix gave an activity recovery of 6.94% and maximum cyclization activity at 60 degrees C, at pH 6.0. The half-time life at 60 degrees C, pH 6.0, in the presence of substrate was 100 min, which was lower than that of the free enzyme. The best activity recovery in this work (6.94%) is approximately five times smaller than that obtained previously using glyoxyl-agarose as support and covalent immobilization. Thus, the best support and method we tested so far for immobilization of CGTase is covalent attachment on glyoxyl-agarose. PMID- 18421599 TI - Improving activity of salt-lyophilized enzymes in organic media. AB - Lyophilization with salts has been identified as an important method of activating enzymes in organic media. Using salt-activated enzymes to transform molecules tethered to solid surfaces in organic phase requires solubilization of enzymes in the solvents. Methods of improving performance of salt-lyophilized enzymes, further, via chemical modification, and use of surfactants and surfactants to create fine emulsions prior to lyophilization are investigated. The reaction system used is transesterification of N-acetyl phenylalanine ethyl ester with methanol or propanol. Initial rate of formation of amino acid esters by subtilisin Carlsberg (SC) was studied and found to increase two to sevenfold by either chemical modification or addition of surfactants in certain solvents, relative to the salt (only)-lyophilized enzyme. The method to prepare highly dispersed enzymes in a salt-surfactant milieu also improved activity by two to threefold. To test the effect of chemical modification on derivatization of drug molecules, acylation of bergenin was investigated using chemically modified SC. PMID- 18421600 TI - Protease production by different thermophilic fungi. AB - A comparative study was carried out to evaluate protease production in solid state fermentation (SSF) and submerged fermentation (SmF) by nine different thermophilic fungi--Thermoascus aurantiacus Miehe, Thermomyces lanuginosus, T. lanuginosus TO.03, Aspergillus flavus 1.2, Aspergillus sp. 13.33, Aspergillus sp. 13.34, Aspergillus sp. 13.35, Rhizomucor pusillus 13.36 and Rhizomucor sp. 13.37- using substrates containing proteins to induce enzyme secretion. Soybean extract (soybean milk), soybean flour, milk powder, rice, and wheat bran were tested. The most satisfactory results were obtained when using wheat bran in SSF. The fungi that stood out in SSF were T. lanuginosus, T. lanuginosus TO.03, Aspergillus sp. 13.34, Aspergillus sp. 13.35, and Rhizomucor sp. 13.37, and those in SmF were T. aurantiacus, T. lanuginosus TO.03, and 13.37. In both fermentation systems, A. flavus 1.2 and R. pusillus 13.36 presented the lowest levels of proteolytic activity. PMID- 18421601 TI - Non-ionic surfactants and non-catalytic protein treatment on enzymatic hydrolysis of pretreated Creeping Wild Ryegrass. AB - Our previous research has shown that saline Creeping Wild Ryegrass (CWR), Leymus triticoides, has a great potential to be used for bioethanol production because of its high fermentable sugar yield, up to 85% cellulose conversion of pretreated CWR. However, the high cost of enzyme is still one of the obstacles making large scale lignocellulosic bioethanol production economically difficult. It is desirable to use reduced enzyme loading to produce fermentable sugars with high yield and low cost. To reduce the enzyme loading, the effect of addition of non ionic surfactants and non-catalytic protein on the enzymatic hydrolysis of pretreated CWR was investigated in this study. Tween 20, Tween 80, and bovine serum albumin (BSA) were used as additives to improve the enzymatic hydrolysis of dilute sulfuric-acid-pretreated CWR. Under the loading of 0.1 g additives/g dry solid, Tween 20 was the most effective additive, followed by Tween 80 and BSA. With the addition of Tween 20 mixed with cellulase loading of 15 FPU/g cellulose, the cellulose conversion increased 14% (from 75 to 89%), which was similar to that with cellulase loading of 30 FPU/g cellulose and without additive addition. The results of cellulase and BSA adsorption on the Avicel PH101, pretreated CWR, and lignaceous residue of pretreated CWR support the theory that the primary mechanism behind the additives is prevention of non-productive adsorption of enzymes on lignaceous material of pretreated CWR. The addition of additives could be a promising technology to improve the enzymatic hydrolysis by reducing the enzyme activity loss caused by non-productive adsorption. PMID- 18421602 TI - Cisplatin-induced hearing loss does not correlate with intracellular platinum concentration. AB - CONCLUSION: Inductively coupled plasma mass spectrometry (ICP-MS) can be applied to organic tissues obtained from experimental animals. Hearing loss does not correlate with the platinum (Pt) concentration found in the inner ear. Drug structure and affinity to inner ear proteins could explain ototoxicity caused by cisplatin. OBJECTIVES: To analyse Pt affinity for brain and ear tissues (of similar embryologic origin) in the Wistar rat and clearance gradient after a single dose, and to correlate these findings with hearing changes. MATERIALS AND METHODS: Thirty-two Wistar rats were intraperitoneally injected with cisplatin at a dose of 5 mg/kg. Animals were sacrificed after obtaining auditory brain responses (ABRs) at 3, 7, 30 and 90 days (nine, seven, seven and nine animals, respectively). Brain and both temporal bones were extracted from each animal and analysed by ICP-MS to determine the absolute concentrations of the metal. Eight non-treated animals were employed as a control group. RESULTS: The ABR thresholds were significantly elevated in animals from all groups after cisplatin treatment. A maximum accumulation of Pt for inner ear and brain was revealed around the first week: 3.175 (57%) and 0.342 (72%), respectively. Pt significantly accumulated in greater quantities in ear than in brain (p<0.01) and was cleared at a higher rate in brain than in ear (p<0.01) following cochlea/brain ratio analysis. No statistically significant correlation was found between amounts of Pt and hearing loss in the study animals. PMID- 18421603 TI - Magnetic resonance imaging of the eustachian tube cartilage. AB - CONCLUSION: No useful examination was available for morphological assessment of the eustachian tube cartilage (ETC) in living subjects, but evaluation of magnetic resonance (MR) imaging enables useful assessment of the eustachian tube (ET) and ETC during preoperative examination before ET surgery. OBJECTIVES: The objective of this study was to investigate the usefulness of MR imaging for analysis and assessment of the ETC. SUBJECTS AND METHODS: Analysis of MR images. RESULTS: The ETC could be demonstrated by oblique parasagittal MR imaging in most subjects, but was poorly defined in others, particularly in aged subjects. The ETC appeared as a hook-shaped structure with wide variability. PMID- 18421604 TI - Involvement of the incudostapedial joint anomaly in conductive deafness. AB - CONCLUSION: The outcome of surgery depends on complexities of middle ear anomalies and definite diagnosis can only be achieved during exploratory tympanotomy. We must be aware that the pathology of the congenital ossicular anomalies is variable and careful surgery is needed for hearing improvement. OBJECTIVE: This study aimed to investigate congenital ossicular malformation. SUBJECTS AND METHODS: Fifteen cases of ossicular anomalies without external ear malformation were studied. The anomaly of the incus-stapes complex was the most frequent. There were two cases of fused incudostapedial (IS) joint, which is an extremely rare occurrence. Case 1 was a 33-year-old man, in whom otosclerosis was suspected and exploratory tympanotomy was performed. The IS joint was fused, and the stapes was immobile. Small fenestra stapedectomy was performed. In case 2, a 52-year-old woman, otosclerosis was also suspected and exploratory tympanotomy was performed. The IS joint was fused and the incus was fixed. After the long process of the incus was cut, the stapes became mobile. However, since the incus remained immobile, it was removed and was placed on the stapes. In both cases, the hearing improved after surgery. The averaged hearing gain of 15 cases was 28.8 dB. PMID- 18421605 TI - Long-term prophylactic treatment of attacks of vertigo in Meniere's disease- comparison of a high with a low dosage of betahistine in an open trial. AB - CONCLUSION: Despite the considerable limitations of an open, non-masked trial, particularly in Meniere's disease (MD), a higher dosage of betahistine dihydrochloride and a long-term treatment seems to be more effective than a low dosage and short-term treatment. OBJECTIVE: To evaluate the prophylactic effects of a low versus high dosage long-term treatment with betahistine-dihydrochloride on the number of attacks in MD. PATIENTS AND METHODS: We performed an open, non masked trial, in which patients with MD received either a low dosage of betahistine-dihydrochloride (16 or 24 mg tid) or a higher dosage of 48 mg tid for at least 12 months. The outcome measure was the number of attacks per month during a 3-month period. Non-parametric tests and a random effects model were used for statistical analysis. RESULTS: A total of 112 patients were included in the analysis: 50 received betahistine-dihydrochloride in a low dosage (16 mg tid, n=21, 24 mg, n=29) and 62 received 48 mg tid. Follow-up examination every 3 months showed that the number of attacks per month decreased in both groups over time. For instance, after 12 months the mean (median) number of attacks dropped from 7.6 (4.5) to 4.4 (2.0) (p<0.0001) in the low-dosage group, and from 8.8 (5.5) to 1.0 (0.0) (p<0.0001) in the high dosage group. The number of attacks after 12 months was significantly lower in the high dosage group than in the low dosage group (p(12M)=0.0002). The treatment was well tolerated in both groups. PMID- 18421606 TI - Tinnitus and brain MRI findings in Japanese elderly. AB - CONCLUSION: There is evidence of an inverse association between cerebral infarction and tinnitus in this study. The effects of cerebral infarction on tinnitus could be explained by a neurophysiological model of tinnitus. OBJECTIVES: We examined the relationship between tinnitus and brain MRI findings including cerebral infarction, brain atrophy, ventricular dilatation, and white matter lesions. SUBJECTS AND METHODS: This was a cross-sectional population-based study of 2193 subjects aged 41-82 years living in Aichi prefecture, Japan. Detailed questionnaires, pure tone audiometry, and brain MRI were performed. RESULTS: After adjusting for potential confounders in a multiple logistic analysis, cerebral infarction was inversely associated with tinnitus (odds ratio (OR)=0.649, 95% confidence interval (CI)=0.477-0.884). Cerebral infarctions of the basal ganglia (OR=0.542), thalamus (OR=0.441), and pons (OR=0.319) were especially associated with tinnitus. Brain atrophy, ventricular dilatation, and white matter lesions had no significant effects on the prevalence of tinnitus. PMID- 18421607 TI - A novel approach to facial nerve decompression: use of Piezosurgery. AB - CONCLUSIONS: The safety of Piezosurgery as regards soft tissues in facial nerve decompression was confirmed. No side effects were detected during unintentional contact with the nerve. OBJECTIVES: The aim of this work was to test Piezosurgery as a new and alternative method to conventional bone tissue management in facial nerve decompression by posterior tympanotomy. PATIENTS AND METHODS: We used Piezosurgery under general anaesthesia on 10 patients affected by facial paralysis following temporal bone fractures. The equipment consists of two hand pieces, two inserts and two peristaltic pumps connected to the control unit. The device uses low frequency ultrasonic waves (24.7-29.5 kHz), the applied power can be modulated between 2.8 and 16 W, and the machine is programmed in accordance with the density of the bone. The micro-vibrations that are created in the piezoelectric hand-piece cause the inserts to vibrate linearly between 60 and 210 microm. RESULTS: Piezosurgery proved effective in sclerotic and pneumatic mastoid. This approach results in significantly less operative blood loss and better visibility in the surgical field compared with conventional methods. Its safety as regards soft tissues was confirmed: no side effects were detected during unintentional contact with the nerve. All patients presented a complete recovery from facial paralysis at the last follow-up. PMID- 18421609 TI - Magnetoencephalography for research of auditory cortex. AB - CONCLUSION: The results could indicate that, during phylogeny and human ontogeny, the central nervous system has enhanced the speech activity from any other activity even though other frequencies could be relevant for survival. OBJECTIVE: People of all ages can experience alterations of auditory perception that progressively increase with aging. The whole approach to these alterations needs not only peripheral (cochlear) or brainstem studies but also an analysis of the auditory cortex. In fact, auditory evoked fields (AEF) may contribute to the understanding of the neural correlate of sound awareness. SUBJECTS AND METHODS: The M100 response after pure tone stimulation (five frequencies ranging from 500 to 8000 Hz) was analyzed in a group of nine adult subjects with normal hearing, older than 25 years of age and under 40 years old. Average M100 field intensity was calculated for all magnetoencephalography (MEG) channels in a 60 ms window around the M100 waveform. RESULTS: The results indicate a more intense cortical response to main speech frequencies (0.5 to 2 kHz) as compared with other frequencies not involved in human conversation. PMID- 18421608 TI - Hearing profile and MRI myelination of auditory pathway in Pelizaeus-Merzbacher disease. AB - CONCLUSIONS: This study showed that delayed auditory pathway myelination is common in Pelizaeus-Merzbacher disease (PMD), but this delay does not necessarily indicate poor hearing function. OBJECTIVE: PMD is a rare recessively inherited X linked leukodystrophy characterized by defective central nervous system myelination owing to a mutation in the proteolipid protein gene (PLP). The aims of this study were to evaluate the hearing function and auditory brain response (ABR) findings of patients with PMD and relate these findings to MRI-assessed myelination in the central auditory pathway. PATIENTS AND METHODS: We retrospectively studied eight male pediatric patients with PMD. Serial auditory examinations included audiometry, behavior audiometry, distortion product otoacoustic emission (DPOAE), and ABR. MRI-assessed myelination in the auditory pathway was evaluated in the PMD patients and in 23 normal young children as a control group. RESULTS: Audiometry showed normal to moderate hearing impairment and the hearing threshold improved with age and became almost normal over time. DPOAEs positivity and only ABR wave I or waves I and II were found in all the patients. MRI showed delayed myelination in all the patients and the auditory pathway was myelinated up to the inferior colliculus in four cases and up to the medial geniculate body in four cases. Serial MRIs showed no progression in myelination. No clear relation was found between hearing threshold and MRI assessed myelination in the auditory pathway. PMID- 18421610 TI - The effect of maxillary sinus surgery on its development. AB - CONCLUSION: The study showed that surgery in the maxillary sinus can affect its development. OBJECTIVE: To quantitatively evaluate the long-term impact of sinus surgery on its development in the rabbit. MATERIALS AND METHODS: This was an experimental study performed at an academic tertiary medical center using 20 4 week-old New Zealand white rabbits. The rabbits underwent unilateral right maxillary sinus surgery. The contralateral maxillary sinus used as a control did not undergo the operation. The maxillary sinus ostium was enlarged on the operated side. Volumetric analysis of the maxillary sinus was performed 1 year post-surgery. The maxillary sinus volumes of both sides were calculated using Multidedector CT and the volumetric measurements of the operated side were compared with the non-operated side. RESULTS: Maxillary sinus development was significantly reduced on the surgical side. Maxillary sinus growth on the surgical side was determined as 87% compared with the non-surgical side. PMID- 18421611 TI - Superselective angiographic embolization for intractable epistaxis. AB - CONCLUSIONS: Routine embolization of the ipsilateral facial artery (FA) is effective because of the high success rate. The use of different embolic materials for the internal maxillary artery (IMA) and the FA was considered safe because of the absence of major complications. OBJECTIVE: To evaluate outcomes of routine embolization of the FA as well as the IMA ipsilateral to the bleeding site for intractable epistaxis, and outcomes using different embolic materials for the FA and the IMA. PATIENTS AND METHODS: Twenty-two patients with intractable epistaxis who underwent superselective embolization were retrospectively analyzed with a mean follow-up of 7 months. The FA and the IMA ipsilateral to the bleeding site were embolized. Two embolic materials, gelatin sponge and microcoils, were used for the IMA and the FA, respectively. RESULTS: The short-term success rate within the first 7 days was 77.3% (17/22). The long term success rate was 95.5% (21/22). There were no major complications in 22 cases. Minor complications occurred in 13 cases (59%). These minor complications usually did not last more than a week and most resolved within a day. PMID- 18421612 TI - Endoscopic management of the inverted papilloma with attachment to the frontal sinus drainage pathway. AB - CONCLUSION: Extensive inverted papilloma (Krouse T3 lesions) with attachment to the frontal sinus drainage pathway can be treated successfully using an endoscopic approach. OBJECTIVES: The endoscopic management of inverted papilloma (IP) has gained in popularity over the last 15 years. However, the appropriate management of lesions involving the frontal sinus and its drainage pathway still has to be determined. PATIENTS AND METHODS: We performed a retrospective review of the results for patients with IP in the Otolaryngology, Head and Neck Surgery Department, Beijing Tongren Hospital from 2004 to 2007 to identify those patients with lesions involving the frontal sinus and its drainage pathway. By their appearance on nasal endoscopic examination and by computed tomography scanning, the tumors were defined using the Krouse staging system. Sinus endoscopy was used to screen for disease after endoscopic resection and the clinical outcomes were analyzed. RESULTS: A total of nine patients (eight males and one female) were identified in which IP involved the frontal sinus and its drainage pathway. Preoperative and postoperative pathological examinations revealed IP as the diagnosis. All tumors were defined as T3 lesions. Four cases with lateral wall of frontal recess tumor attachment underwent an endoscopic Draf IIA frontal sinusotomy. Three cases with either lateral and posterior walls of frontal recess and frontal infundibulum attachment, or medial and posterior walls of frontal recess and frontal infundibulum attachment underwent a Draf IIB procedure. Two cases with posterior wall of frontal recess and frontal infundibulum attachment, or medial, lateral, and posterior walls of frontal recess and frontal infundibulum attachment underwent a Draf III procedure. All patients remain disease-free after an average follow-up period of 15 months. PMID- 18421613 TI - Combination of Bernouilli effect producing maneuver-induced pharyngeal narrowing rate with body mass index as predictive tool for obstructive sleep apnea syndrome. AB - CONCLUSIONS: It is suggested that the combination of the pharyngeal narrowing rate during the Bernouilli effect producing maneuver (BEPM) with the body mass index (BMI) might be a promising predictive method for obstructive sleep apnea syndrome (OSAS). OBJECTIVES: An attempt was made to clarify the possibility that the dynamic narrowing of the pharynx evaluated by nasopharyngoscopy with BEPM, a forced inspiration through the nose with the mouth closed, might be a reliable clinical daytime predictor for identifying patients at risk of developing OSAS. SUBJECTS AND METHODS: Subjects were 57 patients complaining of snoring and sleepiness during daytime. Endoscopic images of the retropalatal pharynx were obtained and their area was measured by NIH imaging. The pharyngeal narrowing rate during quiet nasal breathing and that during BEPM were compared to assess pharyngeal dynamics. To increase the sensitivity of this method, BMI was added to the pharyngeal narrowing rate during BEPM. RESULTS: The criterion of BEPM at a cut-off value of 50% with BMI at a cut-off value of 25 kg/m(2) achieved a sensitivity of 93%, while the specificity was 67% in differentiating simple snorers from OSAS patients (apnea index >5). In addition, the likelihood ratio of the method was found to be 2.81. PMID- 18421614 TI - Analysis of curative effects on laryngeal carcinoma patients in the northeast region of China. AB - CONCLUSIONS: In the past 20 years, the level of laryngeal carcinoma treatment in our country has been significantly improved. Early diagnosis is the key for increasing the ratio of larynx preservation surgeries and improving survival rates. The main causes of death within 5 years are local recurrence and metastasis. OBJECTIVE: To describe the main treatment methods for laryngeal carcinoma in China in the 1980s and 1990s and their prospective effects and investigate the prognostic factors. PATIENTS AND METHODS: A retrospective investigation was performed on the 1115 laryngeal carcinoma patients receiving treatment in the department of ENT of the First Affiliated Hospital of China Medical University during 1983-1996 and the survival rates and causes of death were analyzed statistically. RESULTS: There were 780 patients surviving for more than 5 years, 260 dead patients, and 75 patients lost to follow-up. According to the cumulative survival rate curve, the 5-year survival rate was 77% (94% for stage I, 89% for stage II, 82% for stage III, and 66% for stage IV). Glottic cancer has the highest 5-year survival rate, followed by supraglottic cancer, subglottic cancer, and transglottic cancer. The 5-year survival rate of patients receiving partial laryngectomy was 85%, while the 5-year survival rate of those receiving total laryngectomy was 68%. The leading causes of death within 5 years were local recurrence and metastasis (70%), and the causes of death were unknown in 14% of cases. PMID- 18421615 TI - Useful combination of intra-arterial chemotherapy and radiation therapy for lateral oropharyngeal wall cancer. AB - CONCLUSION: A concomitant treatment of intra-arterial chemotherapy and radiation therapy is a promising therapeutic option for oropharyngeal cancers. OBJECTIVES: Treatment for oropharyngeal cancer has been far from standardized because of its pathophysiologic complexity and its low incidence. In our department, T1 stage tumors with N0 or N1 status are primarily treated surgically, while T1 tumors with N2 or more advanced lymph node involvement are additionally treated with concomitant chemoradiotherapy (CRT). Treatment for T2, T3, and T4 tumors is based on CRT, but surgery is also performed if necessary. PATIENTS AND METHODS: The study included 73 patients with squamous cell carcinomas of the lateral oropharyngeal wall who received first-line therapy at our department between May 1993 and October 2003. RESULTS: The 5-year disease-specific survival by disease stage was 100% for stage I, 90.9% for stage II, 88.2% for stage III, 69.8% for stage Iva, and 22.2% for stage IVb. The overall 5-year disease-specific survival was 71.8%, and the overall 5-year crude survival was 54.1%. PMID- 18421616 TI - Increased viral load correlates with improved survival in HPV-16-associated tonsil carcinoma patients. AB - CONCLUSION: Our results indicate that viral load may serve as an independent prognostic indicator for patients with HPV-16-associated squamous cell carcinoma of the tonsil. OBJECTIVE: HPV-16 has gained increasing attention as a possible causative agent for squamous cell carcinoma of the tonsil. Recent reports have indicated that the viral load within the tumor, along with other factors, may be correlated to the patient survival. In this study, we sought to examine HPV-16 viral load as an independent prognostic indicator. PATIENTS AND METHODS: DNA was extracted from 35 tonsil carcinoma samples and the viral load was determined by real-time PCR. The patients were divided into four groups according to HPV-16 viral load. The correlation between viral load and recurrence, disease-free survival, and overall survival was assessed. RESULTS: We found that HPV-positive patients with the highest viral loads had improved overall and disease-free survival. Recurrences of squamous cell carcinoma were significantly less likely to occur with increasing viral load. PMID- 18421617 TI - Concurrent chemoradiotherapy with cisplatin and fluorouracil for locally advanced hypopharyngeal carcinoma. AB - CONCLUSIONS: The concurrent administration of cisplatin and fluorouracil (CCRT) during radiotherapy after induction chemotherapy had better results than induction chemotherapy followed by radiotherapy alone (CT+RT) for overall survival, laryngeal preservation, and locoregional control in patients with locally advanced hyopharyngeal cancer. OBJECTIVES: To report treatment results comparing CCRT with CT+RT in locally advanced hypopharyngeal cancer. PATIENTS AND METHODS: Sixty-six consecutive patients treated with curative intent were analyzed retrospectively. Thirty-eight patients were treated with induction chemotherapy followed by definitive RT, and 28 patients with induction chemotherapy followed by concurrent chemoradiotherapy. All patients had more than three cycles of continuous infusion of cisplatin and 5-fluorouracil every 3 weeks. The median dose of irradiation was 70 Gy in the CT+RT group and 68.4 Gy in the CCRT group, respectively. RESULTS: The overall 5-year survival rates were 24% for the CT+RT group and 42% for the CCRT group (p=0.031). The 3-year pharyngolaryngectomy-free survival rates were 32% for the CT+RT group and 63% for the CCRT group (p=0.032). The 3-year locoregional control rate for CCRT was significantly better than that for the CT+RT (52% vs 23%, p=0.004). Acute hematologic toxicity appeared in 39% of the CT+RT patients and 75% of the CCRT patients. PMID- 18421618 TI - Sonography-guided OK-432 sclerotherapy for benign thyroid cysts. AB - CONCLUSION: Sonography-guided OK-432 sclerotherapy is effective and safe, and offers a useful alternative treatment for benign thyroid cysts in selected patients. OBJECTIVE: To review the results of sonography guided OK-432 sclerotherapy for benign thyroid cysts. PATIENTS AND METHODS: The details of 20 patients who registered at our institute from December 2002 through August 2006, with recurrent cystic thyroid nodules after two or more aspirations, and who were cytologically negative for malignancy, were reviewed. All cases received OK-432 injections at a concentration of 0.05 mg/ml with sonography. When treatment results were unsatisfactory, the procedure was repeated up to three times at approximately 4 week intervals (mean number, 2.05). RESULTS: Five of the 20 cases were excluded for various reasons. Eleven of the remaining 15 cases (73.3%) showed good responses (<10 mm in diameter), whereas the other 4 patients showed poor responses. All patients tolerated the therapy well, and no significant morbidity or complication occurred among patients treated by OK-432 sclerotherapy. PMID- 18421619 TI - Frontline providers harbor misconceptions about the bedside evaluation of dizzy patients. PMID- 18421620 TI - Identification of human cytochrome P450 enzymes involved in the metabolism of IN 1130, a novel activin receptor-like kinase-5 (ALK5) inhibitor. AB - 1. The in vitro metabolism of 3-((5-(6-methylpyridin-2-yl)-4-(quinoxalin-6-yl)-1H imidazol-2-yl)methyl)benzamide (IN-1,130), a selective activin receptor-like kinase-5 (ALK5) inhibitor and a candidate drug for fibrotic disease, was studied. 2. The cytochrome P450s (CYPs) responsible for metabolism of IN-1,130 in liver microsomes of rat, mouse, dog, monkey and human, and in human CYP supersomestrade mark, were identified using specific CYP inhibitors. The order of disappearance of IN-1,130 in various liver microsomal systems studied was as follows: monkey, mouse, rat, human, and dog. 3. Five distinct metabolites (M1-M5) were identified in all the above microsomes and their production was substantially inhibited by CYP inhibitors such as SKF-525A and ketoconazole. Among nine human CYP supersomestrade mark examined, CYP3A4, CYP2C8, CYP2D6 1, and CYP2C19 were involved in the metabolism of IN-1,130, and the production of metabolites were significantly inhibited by specific CYP inhibitors. IN-1,130 disappeared fastest in CYP2C8 supersomes. CYP3A4 produced four metabolites of IN-1,130 (M1-M4), whereas supersomes expressing human FMO cDNAs, such as FMO1, FMO3, and FMO5, produced no metabolites. 4. Hence, it is concluded that metabolism of IN-1,130 is mediated by CYP3A4, CYP2C8, CYP2D6 1, and CYP2C19. PMID- 18421621 TI - Induction of cytochrome P450s by terpene trilactones and flavonoids of the Ginkgo biloba extract EGb 761 in rats. AB - 1. Ginkgo biloba is one of the most popular herbal medicines worldwide due to its memory-enhancing and cognition-improving effects. The current study was designed to investigate the effects of five major constituents (bilobalide, ginkgolide A, B, quercetin, and kaempferol) in the standardized G. biloba extract EGb 761 on various cytochrome P450s (CYPs) in rats. 2. The activity of CYP450 was measured by the quantification of six metabolites from multiple cytochrome P450 probe substrates using a validated liquid chromatography coupled with tandem mass spectrometry detection (LC-MS/MS) method. The levels of messenger RNA (mRNA) and protein of various CYPs were determined by reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blotting analysis, respectively. 3. Bilobalide significantly induced the activity, protein, and mRNA expression of CYP3A1 and 1A2, and increased CYP2E1 activity and CYP2B1/2 protein expression in a dose dependent manner. 4. Ginkgolide A, B, quercetin, and kaempferol did not affect CYP3A1, but induced CYP1A2 in a dose-dependent manner. EGb 761 and the five individual constituents had no effects on rat CYP2D2, 2C11 and 2C7. 5. The results indicate that bilobalide, and to a lesser extent ginkgolide A, B, quercetin, and kaempferol, play a key role in the effects of EGb 761 on CYP induction. Further study is needed to elucidate the mechanism of CYP3A induction by EGb 761 and bilobalide. PMID- 18421622 TI - Prediction of metabolic clearance of diclofenac in adjuvant-induced arthritis rats using a substrate depletion assay. AB - 1. The purpose of this study was to evaluate drug clearance measured by the metabolic intrinsic clearance (CL(int)) in a substrate depletion assay in comparison with the in vivo clearance (CL(tot)) observed in adjuvant-induced arthritis (AA) rats. 2. After intravenous administration of diclofenac as a model drug, CL(tot) was 2.8-fold higher in AA rats than in control rats. In two different substrate depletion assays with liver microsomes for glucuronidation and hydroxylation, the CL(int) values for glucuronidation was significantly decreased in AA rats to 60% of the value in control rats, whereas the CL(int) values for hydroxylation were similar. The unbound fraction of diclofenac in plasma (f(u, plasma)) was significantly higher (2.8-fold) in AA rats than in control rats. 3. Hepatic clearance predicted from the CL(int) values for both biotransformation pathways and f(u, plasma) was higher in AA rats than in control rats, with good consistency between predicted and observed values. The same results were obtained for experiments using hepatocytes. 4. The plasma protein binding activities, rather than metabolic clearance, in both types of rats would be a determining factor in the pharmacokinetic behaviour differences between control and AA rats. 5. In summary, substrate depletion assays with liver microsomes and hepatocytes in combination with protein binding assessment can help to predict changes in pharmacokinetics under AA conditions. PMID- 18421623 TI - In vitro drug-drug interaction studies with febuxostat, a novel non-purine selective inhibitor of xanthine oxidase: plasma protein binding, identification of metabolic enzymes and cytochrome P450 inhibition. AB - 1. The potential for drug-drug interactions with febuxostat was examined in the following three in vitro systems: the characteristics of the binding of febuxostat to human plasma proteins; identification of the cytochrome P450 (CYP) and UDP-glucuronosyltransferase (UGT) enzymes participating in the metabolism of febuxostat; and the potential inhibitory effects of febuxostat on typical CYP reactions. 2. The results have shown that the presence of ibuprofen or warfarin did not change the plasma protein binding of febuxostat, and that febuxostat did not influence the plasma protein binding of ibuprofen or warfarin. These results indicate that there is little possibility that febuxostat causes a drug-drug interaction by binding to albumin. 3. The UGT 1 and 2 families were involved in the glucuronidation, and several CYPs participated in the metabolism of febuxostat, suggesting that there is little possibility that the blood concentration of febuxostat varies widely even if febuxostat is concomitantly administered with drugs that inhibit CYP or UGT enzyme. Examination of the inhibitory effect of febuxostat on CYP enzymes suggests that febuxostat minimally inhibits the activities of any CYP. 4. The results demonstrate that febuxostat is a novel anti-hyperuricaemia drug with low drug-drug interaction potential in clinical use. PMID- 18421624 TI - Pharmacokinetics and metabolism of KW-4490, a selective phosphodiesterase 4 inhibitor: difference in excretion of KW-4490 and acylglucuronide metabolites between rats and cynomolgus monkeys. AB - 1. The pharmacokinetics and metabolism of KW-4490, a selective phosphodiesterase 4 inhibitor, were investigated in rats and monkeys. After oral administration, KW 4490 was rapidly absorbed, and then its plasma concentrations apparently declined with half-lives of approximately 5 h in rats and 3.5 h in cynomolgus monkeys; however, a number of secondary peaks were apparent in the profiles for both species. The plasma pharmacokinetics of KW-4490 were comparable between rats and monkeys. 2. After oral administration, KW-4490 was mainly eliminated by metabolism to acylglucuronides and renal excretion in the unchanged form. KW-4490 acylglucuronides were found in monkey but not rat urine. In rats, KW-4490 acylglucuronides were excreted only in bile. Although the pathway of excretion of acylglucuronides differed between rats and monkeys, cumulative excretion in the two animals was very similar, as expected from comparable hepatic clearance for glucuronidation in rat and monkey liver microsomes. 3. The glomerular filtration rate of unbound KW-4490 indicated that renal tubular secretion was significant in monkeys, whereas reabsorption was significant in rats. These species differences in urinary excretion of KW-4490 and its acylglucuronide metabolites are most likely due to substrate specificity of active transporters in rat and monkey kidney. PMID- 18421625 TI - Comparative evaluation of absorption, distribution, and excretion of YM758, a novel If channel inhibitor, between albino and non-albino rats. AB - 1. YM758 is a novel If channel inhibitor for the treatment of stable angina and atrial fibrillation. The absorption, distribution, and excretion of YM758 have been investigated in albino and non-albino rats after a single oral administration of (14)C-YM758 monophosphate. 2. YM758 was well absorbed from all segments of the gastrointestinal tract except for the stomach. After oral administration, the ratio of AUC(0-1 h) between the plasma concentrations of radioactivity and the unchanged drug was estimated to be 17.7%, which suggests metabolism. 3. The distribution of the radioactivity derived from (14)C-YM758 in tissues was evaluated both in albino and non-albino rats. The radioactivity concentrations in most tissues were higher than those in plasma, which indicates that the radioactivity is well distributed to tissues. Extensive accumulation and slower elimination of radioactivity were noted in the thoracic aorta of albino and non-albino rats as well as in the eyeballs of non-albino rats. The recovery rates of radioactivity in urine and bile after oral dosing to bile duct cannulated albino rats were 17.8% and 57.3%, respectively. 4. These results suggest that YM758 was extensively absorbed, subjected to metabolism, and excreted mainly into the bile after oral administration to rats, and extensive accumulation of the unchanged drug and/or metabolites into tissues such as the thoracic aorta and eyeballs was observed. PMID- 18421626 TI - Metabolism of prazosin in rat and characterization of metabolites in plasma, urine, faeces, brain and bile using liquid chromatography/mass spectrometry (LC/MS). AB - 1. Prazosin, 2-[4-(2-furanoyl)-piperazin-1-yl]-4-amino-6,7-dimethoxyquinazoline, is an antihypertensive agent that has been used safely since 1976 and is currently being investigated for the treatment of post-traumatic stress disorder. The in vivo metabolism of prazosin in rat was first reported in 1977, although at the time analytical techniques were not as sophisticated, nor were the mass spectrometers as sensitive, as today. Recently, the in vitro metabolism of prazosin in rat liver microsomes and cryopreserved hepatocytes was investigated using liquid chromatography/mass spectrometry (LC/MS), which revealed new metabolic pathways. 2. In the present work, rat in vivo metabolism was reinvestigated using a quadrupole time-of-flight mass spectrometer coupled with ultra-performance liquid chromatography, or chip-based nanoflow electrospray ionization, with the aim of identifying metabolites revealed by the in vitro studies and any new metabolites. 3. It is reported that prazosin was metabolized in rats to produce the metabolites observed in vitro. In addition, new phase I metabolites, M18, M20 and M21, were formed and conjugation with glucose or taurine formed the new phase II metabolites, M16 and M19, respectively. 4. Evidence for bioactivation of prazosin included detection of ring-opened metabolites (M4 and M7) and a cysteinyl-glycine conjugate (M17). Further support to the structure of the ring-opened metabolite M7 was obtained by nuclear magnetic resonance (NMR) experiments on M7 isolated from urine. PMID- 18421627 TI - Expanded vs. equal interval spaced retrieval practice: exploring different schedules of spacing and retention interval in younger and older adults. AB - The present study was designed to help answer several questions regarding the impact of spacing and expanded retrieval on memory performance in younger and older adults. Three expanded/equal interval schedule pairings, matched in average spacing (1-2-3/2-2-2; 1-3-5/3-3-3; and 1-3-8/4-4-4), were compared, and the effect of retention interval on spaced retrieval benefits was examined by comparing performance on a same day test to a test delayed by 24 h. Both age groups showed a learning phase retrieval success advantage for expanded items compared to equal interval items. Only older adults in the same day test condition showed a significant expansion effect in final recall. After a 24-h delay, the final recall advantage for items in the expanded condition was lost in both groups, and in fact these items were at a significant recall disadvantage for younger adults. Results indicate that younger and older adults benefit from a rehearsal technique that incorporated any type of spaced retrieval whether it is distributed as an expanding schedule or not. Although we did not find robust advantages for expanded retrieval compared to equal interval practice, there could be certain advantages (such as reinforcement due to high success rates) to using expanded retrieval depending on the ultimate goals of an individual memory training program. PMID- 18421628 TI - Effects of item difficulty on the retrieval of solutions during cognitive skill acquisition: age differences. AB - In cognitive skill learning, shifts to better strategies for obtaining solutions often occur while associations between problems and solutions are being strengthened. In two skill learning experiments, we examined the effects of item difficulty on the retrieval of solutions and the learning of problem-solution associations in younger and older adults. The results of both experiments demonstrated an 'easy effect' in both younger and older adults, such that the retrieval of solutions as well recognition memory for problems was best for the easier items. In addition, a 'hard effect' was found in younger adults, but not in older adults, whereby the retrieval of solutions as well as recognition memory for problems was better for harder items than for medium-difficulty items. The finding that increased computational demands at the item level delayed item memorization and the retrieval of solutions in older adults but not younger adults is consistent with a general-resources account of age-related differences in skill learning. PMID- 18421629 TI - White matter hyperintensities and working memory: an explorative study. AB - White matter hyperintensities (WMH) are commonly observed in elderly people and may have the most profound effect on executive functions, including working memory. Surprisingly, the Digit Span backward, a frequently employed working memory task, reveals no association with WMH. In the present study, it was investigated whether more detailed analyses of WMH variables and study sample selection are important when establishing a possible relationship between the Digit Span backward and WMH. To accomplish this, the Digit Span backward and additional working memory tests, WMH subscores, and cardiovascular risk factors were examined. The results revealed that performance on the Digit Span backward test is unrelated to WMH, whereas a relationship between other working memory tests and WMH was confirmed. Furthermore, a division between several white matter regions seems important; hyperintensities in the frontal deep white matter regions were the strongest predictor of working memory performance. PMID- 18421631 TI - The pearls and pitfalls of skull trephination as described in the Hippocratic treatise "On Head Wounds". AB - The purpose of our current study is to describe the guidelines regarding trephination as suggested in the Hippocratic writer's book, "Peri ton en cephali traumaton" ("On Head Wounds"). The ancient Greek text was reviewed as well as two English translations. The Hippocratic author described the indications, timing, and techniques of trephination for patients with head injury. He emphasized that attention should be paid to the details of the employed technique. He also commented on the difference of skull thickness at different anatomical sites and also between different age groups. The Hippocratic recommendations provided details for the performance of trephination by the ancient Greek physicians that are still considered important in modern neurosurgery. PMID- 18421632 TI - Warts and the kings of Parthia: an ancient representation of hereditary Neurofibromatosis depicted in coins. AB - Parthian coins depict a nodule on the face of many of their kings over succeeding generations. Loosely described as a wart in the literature, the nature of these lesions has been the subject of speculation. The accepted view is that they were unlikely to be simply a cosmetic or symbolic feature. Evidence suggests that they may represent the cutaneous tumors of Neurofibromatosis. The hereditary nature and physical appearance of these round nodules are consistent with this diagnosis. Although final proof may be lacking, these prominent facial features are worthy of discussion even though the matter may not be settled with certainty. PMID- 18421633 TI - Migraine in Gilbertus Anglicus' Compendium medicinae. The cases of MS Sloane 3486 and Wellcome MS 537. AB - Migraine is a common neurological disorder that has deserved attention since Antiquity. The English physician Gilbertus Anglicus dedicated a separate chapter of his thirteenth-century Compendium medicinae to it, which was then reproduced in some medical manuscripts from the later Middle Ages. MS Sloane 3486 and Wellcome MS 537 are two extant Middle English translations of the Latin exemplar composed by the end of the fifteenth century. This work will analyze how their copyists reduced Anglicus' long theoretical dissertations on the aetiology and semiology of migraine, in order to emphasize the real therapeutic needs of a nonuniversity-trained audience. PMID- 18421634 TI - The experiments of Ramon M. Termeyer SJ on the electric eel in the River Plate region (c. 1760) and other early accounts of Electrophorus electricus. AB - This paper focuses on Ramon M. Termeyer SJ (1737-1814?), a naturalist who experimented with the electric eel in the River Plate region during the 1760s. After going through an enumeration of the chroniclers that since the sixteenth century noticed the benumbing discharge of Electrophorus electricus, the article summarizes the work that immediately preceded Termeyer's and considers as a term of comparison the experiments on the electric eel performed by Bertrand Bajon (fl. 1751-1778) in the French Guyanne. It ends by discussing the meaning of Termeyer's 1781 and 1810 articles in the light of contemporary ideas of animal electricity. PMID- 18421635 TI - Cerebral localization in the Netherlands in the nineteenth century: emphasizing the work of Aletta Jacobs. AB - This paper gives an overview of the ideas on cerebral localization as they were formulated in the Netherlands in the nineteenth century. In general, it appears that, given the discussions in France, Germany, and Britain, surprisingly little was written and hardly any serious research was performed. Special attention is given to the dissertation on the localization of sensory-motor functions by Aletta Jacobs (1854-1929), known for her social and political activities to improve the position of women. Although it was not a particularly thorough study, it was the only one that seriously and empirically addressed the topic of localization. PMID- 18421636 TI - Science and fate: Lina Stern (1878-1968), a neurophysiologist and biochemist. AB - Lina Stern (1878-1968), a neurophysiologist and biochemist, was born in Russia. She studied at the University of Geneva, Switzerland, where, after graduating, she conducted original research in physiology and biochemistry. In 1918, Stern was the first woman to be awarded a professional title at the University of Geneva and headed the department of Physiological Chemistry. She is deservedly considered to be one of the first scientists to entertain the concept of a blood brain barrier. In 1929, Stern founded the Institute of Physiology in Moscow, of which she was director until 1948, when it was discontinued. Under her leadership, multidisciplinary groups of colleagues worked on the problems of the blood-brain and tissue-brain barriers and homeostasis of the brain. In 1939, Stern was elected full member of the Academy of Sciences and became its first female member ever. Most scientists manage to conduct their research by adjusting to the political and social situations surrounding them. Lina Stern did not follow this path. This small woman of complete devotion to science took the drastic decisions that altered her life. Though destiny was not kind to her, Lina Stern did not compromise. Despite a threat of execution, prolonged imprisonment, and exile she was never broken as a scientist and always maintained her dignity. PMID- 18421637 TI - Varieties of publication patterns in neuroscience at the cognitive turn. AB - The quantification of publication activity and impact has become a key element in the evaluation of scientific excellence. However, it is unclear to what extent this grasps the diversity of research communication that accompanies the transition of scientific fields. This contribution investigates number, categorization, and impact of publications (i.e., publication patterns) of six scientists active at the cognitive turn, which promoted the information processing perspective on neuronal processes in different communities: Horace Barlow, Theodore Bullock, Ralph Gerard, Donald MacKay, Warren McCulloch, and Werner Reichardt. The large variety of publication patterns revealed indicates the limits of standardized evaluation procedures based on publication activity. PMID- 18421638 TI - The evolution of the terminology of the basal ganglia, or are they nuclei? AB - Basal ganglia have been generally used to refer to some subcortical nuclei. However, it is a misnomer since ganglion is a group of nerve cells especially located outside of the brain or spinal cord. We evaluated the terminology of the basal ganglia from historical and terminological points of view. PMID- 18421639 TI - Twelfth annual meeting of the International Society for the History of the Neurosciences. June 19-23, 2007. AB - Christian Baumann (Justus-Liebig-University, Giessen, Germany): Wilbrand's ideas of the visual cortex. Hermann Wilbrand (1851-1935) is considered one of the founders of neuro-opthalmology. He is best known for the monumental handbook, Die Neurologie des Auges (Wilbrand &Saenger, 1898-1922). Prior to this encyclopedic work, Wilbrand published three clinical monographs on the diagnosis of brain diseases with the help of ophthalmological examinations(Wilbrand 1881, 1884, 1890). But Wilbrand not only treated clinical aspects but also supplied evidence for the localization of the optical center in the calcarine fissure of the occipital cortex. Moreover, he worked out theories of the organization of the visual cortex that, as he postulated, must contain subdivisions corresponding to the qualities of visual sensation such as light, form, and color. Wilbrand also considered the binocular input of the visual cortex and put forward a detailed scheme of the projection of the two retinae to the occipital cortex that anticipated modern concepts of ocular dominance columns. His ideas are critically reviewed in the light of current opinions about his topics. PMID- 18421640 TI - Dermo-optical perception: the non-synesthetic "palpability of colors" a comment on Larner (2006). AB - We comment on Larner's (2006) recent description of the seventeenth-century case of a blind man who could differentiate the color of objects by touch. This ability is generally known as "dermo-optical perception" and is due to the cutaneous temperature sense rather than to synesthetic processing. Although devoid of references to the phenomenon of dermo-optical perception, Larner's communication is highly valuable because it raises several issues relevant to present-day neurosciences. These comprise functional reorganization after sensory loss, handedness effects, and differences between single fingers in the sensitivity to thermal changes. PMID- 18421641 TI - Can the irrelevant speech effect turn into a stimulus suffix effect? AB - The irrelevant sound effect (ISE) and the stimulus suffix effect (SSE) are two qualitatively different phenomena, although in both paradigms irrelevant auditory material is played while a verbal serial recall task is being performed. Jones, Macken, and Nicholls (2004) have proposed the effect of irrelevant speech on auditory serial recall to switch from an ISE to an SSE mechanism, if the auditory perceptive similarity of relevant and irrelevant material is maximized. The experiment reported here (n = 36) tested this hypothesis by exploring auditory serial recall performance both under irrelevant speech and under speech suffix conditions. These speech materials were spoken either by the same voice as the auditory items to be recalled or by a different voice. The experimental conditions were such that the likelihood of obtaining an SSE was maximized. The results, however, show that irrelevant speech - in contrast to speech suffixes - affects auditory serial recall independently of its perceptive similarity to the items to be recalled and thus in terms of an ISE mechanism that crucially extends to recency. The ISE thus cannot turn into an SSE. PMID- 18421642 TI - A comparison of backward and forward spatial spans. AB - The standard Corsi blocks task is frequently used to measure the capacity of visuospatial working memory, but the implications of the use of both forward and backward recall are still unclear. In the present study, we showed that the backward Corsi task is particularly powerful in discriminating between low- and high-spatial-ability individuals and involves different processes from those involved in the forward task. From a sample of 425 participants we selected one group of 20 high-spatial-ability participants and one of 20 low-spatial-ability participants. The results demonstrated that a backward spatial span offers specific information not available from a forward spatial span, and that there was no facilitation due to a descending format. In particular, in the low-spatial ability group, performance was generally poorer, but backward Corsi recall was lower than forward recall, and participants did not show any advantage following the descending presentation format - which in some contexts is considered to reduce proactive interference. We conclude that the backward Corsi task has specific value and that the assumption of fully parallel verbal and visuospatial working-memory systems can lead to a variety of misunderstandings. PMID- 18421643 TI - Sex differences in mental rotation with polygons of different complexity: Do men utilize holistic processes whereas women prefer piecemeal ones? AB - Sex differences in mental rotation were investigated as a function of stimulus complexity with a sample size of N = 72. Replicating earlier findings with polygons, mental rotation was faster for males than for females, and reaction time increased with more complex polygons. Additionally, sex differences increased for complex polygons. Most importantly, however, mental rotation speed decreased with increasing complexity for women but did not change for men. Thus, the sex effects reflect a difference in strategy, with women mentally rotating the polygons in an analytic, piecemeal fashion and men using a holistic mode of mental rotation. PMID- 18421644 TI - [Results in otosurgically treated patients with acoustic neuroma. Part 2: Hearing results after middle fossa approach]. AB - BACKGROUND: The surgical therapy of acoustic neuromas has been seen critically lately. Alternative methods like the gamma-knife treatment open a wide field of discussion. PATIENTS: The results of 392 patients are presented operated on via a middle fossa approach between 1989 and 2004. RESULTS: Hearing preservation according to the 50:50 rule (hearing loss less than 50 dB in pure tone audiogram 500-1000-2000 Hz, speech discrimination>50%) was possible in 59.7% of patients. A hearing impression up to 90 dB was recognized in 83.1%, 16.9% of cases were deaf. There was no difference between tumors confined to the inner ear canal and tumors extending into the cerebello pontine angle. There was also no difference between tumors up to a volume of 100 mm3 and tumors between 100 mm3 and 500 mm3. CONCLUSION: Acoustic neuroma confined to the internal meatus or with little extend into the cerebello pontine angle (i. e. no contact to structures of the brain stem or vessels of the posterior fossa) are feasible for resection via the middle fossa approach. The possibility of hearing preservation and low morbidity makes it the treatment of choice for this group of tumors. PMID- 18421645 TI - [Imaging of Cochlear Structures by Optical Coherence Tomography (OCT). Temporal bone experiments for an OCT-guided cochleostomy technique]. AB - BACKGROUND: Optical Coherence Tomography (OCT) is a relatively new imaging technique, which provides scans similar to sonography on an optical base. We questioned whether OCT may be helpful in optimizing anatomical orientation in cochlear implant (CI) surgery. In a study on temporal bone specimens we tested, whether OCT provides information about the cochlear topography, particularly in situations, when for cochleostomy the bony otic capsule is already opened but the membranous endosteal layer is still intact. MATERIAL AND METHODS: OCT was performed on five human temporal bone preparations, in which the cochleostomy was carried out still leaving the endosteum covering the fluid-filled inner ear intact. A prototype of operating microscope was used, in which a spectral-domain OCT (SD-OCT) with a central wavelength of 840 nm was integrated. RESULTS: On all scans, OCT supplies information about inner ear structures, such as the lateral attachment of the basilar membrane dividing the scalae. Even delicate structures like the Reissner's membrane could be identified in one case. CONCLUSIONS: This pilot study clearly documents the possibility to identify inner ear structures, especially the site of the scala tympani while its enveloping membranes are still intact. These findings may have an impact on cochlear implant surgery, especially as an orientation guide to localize the scala tympani precisely before opening the fluid-filled inner ear. PMID- 18421646 TI - [Articulation deficits in children with hearing impairment and children with specific language impairment - a matched-pairs-study]. AB - BACKGROUND: Children with hearing impairment commonly present with articulation deficits. This is considered one main characteristic. METHODS AND PATIENTS: 24 pairs of children (5;00 - 6;11 yrs) with articulation deficits in the context of a language disorder were matched in the severity of the language disorder, age, gender and nonverbal intelligence. Of each matched pair one child had a hearing impairment, the other a specific language impairment. Both groups were compared for their articulation. In children with hearing impairment the influence of the severity of the hearing loss was evaluated. RESULTS: In both groups the same type of articulation deficit for consonants was found. Both groups of children had most frequently problems with sibilants. Children with specific language impairment had slightly more severe articulation deficits. In unvoiced plosives children with specific language impairment demonstrated more often substitutions. The severity of the language disorder, but not the severity of the articulation deficit, was correlated with the degree of hearing loss. CONCLUSION: Articulation deficits in children with hearing impairment are slightly less than in children with specific language impairment, but follow the same pattern. PMID- 18421647 TI - [Results in otosurgically treated patients with acoustic neuroma. Part 1: Facial nerve function after translabyrinthine and middle fossa resection]. AB - BACKGROUND: Regardless of all efforts to preserve a residual hearing, facial nerve function is still the primary issue for patients with acoustic neuromas. Since alternative methods like the gamma-knife treatment are of increasing importance, results after surgery have to be compared and discussed critically. PATIENTS: The results of 538 patients operated at the Dept. of Otolaryngology, Head and Neck Surgery, University of Wuerzburg between 1989 and 2004 are presented. 392 had surgery via a middle fossa, 146 via a translabyrinthine approach. RESULTS: Shortly after surgery 82.2 % of patients showed no facial palsy. 74 patients had an incomplete, 7 a complete paresis. After 12 months 144 of the patients operated on via middle fossa approach showed no palsy, 13 had an incomplete and one a complete paresis. Out of 76 patients operated on translabyrinthine approach 75 had no paresis, one had an incomplete, and no patient had a complete paresis. There was no difference between intrameatal tumors and tumors extending beyond the porus. Regular facial nerve function was seen in 93 %, postoperatively. Permanent paresis was seen in 0.4 % of cases. CONCLUSION: Acoustic neuroma confined to the internal meatus or with minor extension into the cerebello-pontine angle (i. e. no contact to structures of the brain stem or vessels of the posterior fossa) are feasible for resection via a middle fossa or translabyrinthine approach. The possibility of hearing preservation combined with low morbidity and good results of facial nerve function makes these approaches the treatment of choice for this group of tumors. PMID- 18421648 TI - [Hybrid operation for a distal aortic arch aneurysm causing left recurrent nerve palsy - Ortner's syndrome]. AB - BACKGROUND: A left recurrent nerve palsy caused by left atrium hypertrophy, dilatation of the left pulmonary artery or a thoracic aortic arch aneurysma is named Ortner's syndrome. CASE REPORT: We report the case of a 84-year old male patient with a posttraumatic aneurysm of the distal aortic arch known for 20 years, which became symptomatic by new presentation of an acute hoarseness caused by a left sided recurrent nerve palsy due to rapid aneurysma expansion. Because of the aneurysma localization, the advanced age and the patient?s morbidity he was treated by a hybrid procedure with an extrathoracic reconstruction of the supraaortic arteries by a right to left carotid-carotid bypass and transposition of the left subclavian artery followed by endovascular implantation of aortic stent grafts for aneurysma exclusion. The left recurrent nerve palsy remained unchanged until now. CONCLUSIONS: Aneursms of the distal aortic have to be considered as a rare cause of a left-sided recurrent nerve palsy. Recent advances in endovascular therapy combined with supraaotic vascular reconstruction offers a less invasive therapeutic option for some of these patients. PMID- 18421649 TI - ["Migration and health"--a subject of high priority]. PMID- 18421650 TI - [Depression, anxiety and posttraumatic stress disorders in labor migrants, asylum seekers and refugees. A systematic overview]. AB - BACKGROUND: In 2006 there were about 200 millions of transnational "voluntary" migrants like labor migrants and "involuntary migrants" like refugees and asylum seekers worldwide. Depression, anxiety and posttraumatic stress disorder (PTSD) is the most prevalent psychiatric disorders in general populations and is reported to be highly prevalent among migrants. AIMS: We aimed to assess and compare syndromes and symptoms of depression and anxiety in labor migrants and refugees; and to examine whether the prevalence rates are associated with study methods' and study quality. METHODS: We systematically searched in the databases MEDLINE and EMBASE for studies published from 1994 - 2007. Studies fulfilling the inclusion criteria are 1) systematically described: 2) and evaluated with 15 quality criteria. RESULTS: The literature search generated 348 results; and 37 fulfilled our inclusion criteria (35 populations) with n = 24 681 migrants (labor migrants: n = 16 971; refugees: n = 7710). Size of studies varies from a minimum of n = 55 participants to a maximum of n = 4558 participants (Median: n = 338). Prevalence rates for depression vary between 3 % and 47 % (labor migrants) and between 3 % and 81 % (refugees); for anxiety between 6 % and 44 % (labor migrants) and between 5 % and 90 % (refugees) and for PTSD between 4 % and 86 %. No study fulfilled all 15 quality criteria. CONCLUSION AND OUTLOOK: Migrants are a heterogeneous group and prevalence rates vary widely between studies. There is a need of high-quality representative studies on migrants' mental health to adequately plan health and social care. PMID- 18421651 TI - [Mental health care for migrants]. AB - BACKGROUND: Global migration and the increasing number of migrants to Europe and Germany diversify the needs in the psychosocial and health care system. Migrants are a heterogeneous group as regards their country of emigration, reasons for migration and legal status. AIMS: We aim to give an overview on 1) mental health of migrants in Germany, 2) cultural associated explanatory of addictive behaviour, 3) utilisation and help-seeking behaviour migrants with particular regard to addicted migrants, and on 4) barriers within the psychosocial care system. RESULTS: Studies on migration, mental health and utilisation of psychosocial institutions especially of institutions for addicted persons show inconsistent results. The results may be conflicting because of the methods used (e. g. small sample size, variety of methods, studies on clinical populations, studies without control-groups, mono-ethnic studies) or because of differences between populations. Therefore, the comparability of results is limited. Migrants use health and psychosocial care institutions differently from non-migrants. Barriers within the psychosocial care system may be caused by uncertainty of learned behaviour of members of staff how to treat migrants or by institutional barriers. CONCLUSION: Our findings show that empirical studies on mental health of migrants are still rare. Further specific investigations are needed to get an in-depth understanding of migrants' mental health and their pattern of psychosocial and health care utilisation to modify responsiveness of services. PMID- 18421652 TI - [Epidemiological perspectives of migration research: the example of cancer]. AB - Epidemiologic research on migration and health in Germany is increasingly focusing on chronic diseases. Migrant studies can help to identify causes of diseases. They also help to identify health inequalities and can thus contribute to improvements in health care. With regard to cancer, descriptive studies among Turkish migrants and ethnic German immigrants from the Former Soviet Union have shown overall low, but slowly increasing cancer rates as compared to autochthonous Germans. For individual cancer sites such as stomach cancer, migrants appear to experience higher risks than the comparison population. Epidemiologic approaches studying cancer care and services for migrants, as well as analytic studies that allow assessing the particular temporal dynamics of cancer risks among migrant groups, are scarce in Germany. Thus, major challenges for cancer epidemiology among migrants exist in Germany. PMID- 18421653 TI - [Intercultural aspects of medical care for undocumented migrants]. AB - In view of the cultural diversity in German society today, the time has long since come when medical care must adjust to its new clientele. This article provides an overview for doctors, medical personnel and psychologists of approaches, backgrounds and networks of migration to Germany, in particular over the little known undocumented migration. This migration has steadily increased in recent years. The author deals with the circumstances which create psychological problems for migrants and what happens when migrants living in this shadow world fall ill. In addition, the article offers an agenda for interculturally competent action in caring for documented and undocumented migrants. Dimensions of cultural differences such as collectivism versus individualism (most of the countries of origin of these migrants in Germany with or without documents are collectivistic) are explained along with differences in styles of communication. The following styles with their impact in actual practice are analyzed: indirect versus direct communication; emotional control versus expressiveness; functionalism versus relationship orientation. PMID- 18421654 TI - [Anxiety and depression by Polish and Vietnamese migrants in Leipzig depending on their adaptation process]. AB - Although the fact that migrants show a lower psychological health status than non migrants is clearly demonstrated by a lot of studies, the connection between migration conditions and psychological health status is seldom analyzed. To close this lack of research 140 Polish and 82 Vietnamese migrants in Leipzig were examined by the anxiety- and depression scale of the HADS and more over asked questions about their cognitive, social, occupational, identificative assimilation status and their perceived discrimination. Both migrant groups showed a higher level of depressive and anxious symptoms than Germans. In regression analyzes only social assimilation and perceived discrimination had a significant influence, language skills and structural (occupational) assimilation could not explain depression or anxiety, but there were differences in regression models between both groups. Beside problems in operationalization, different migrant biographies and structural conditions have to be discussed as reason for these results. PMID- 18421655 TI - [Traumatic events and posttraumatic stress disorder in Turkish-speaking patients in primary care]. AB - In a consecutive cross-sectional study 195 Turkish/Kurdish patients from seven general practices (40.5 +/- 13.3 years, 63.6 % female) were examined with regard to traumatisation and psychological distress with the Essen Trauma Inventory (ETI) and Hospital Anxiety and Depression Scale (HADS). In mean the average duration of living in Germany was 25,3 years (SD = 8.9). 53.3 % reported at least one traumatic event in their life. Of those who experienced a traumatic event, 19.6 % met testpsychometric a positive diagnosis of PTSD. In general, patients with PTSD were more likely to report more about doctor visits (p 0.993), and performed better than decision rules based on the signal-to-noise-ratio (AUC<0.934) or the estimated Cramer-Rao-bound on the errors of a spectral fitting (AUC<0.952). This probabilistic assessment of the data quality provides comprehensible confidence images and allows filtering the input of any subsequent data processing, i.e., quantitation or pattern recognition, in an automated fashion. It thus can increase robustness and reliability of the final diagnostic evaluation and allows for the automation of a tedious part of MRSI data analysis. PMID- 18421693 TI - Oxygen-enhanced MR imaging of mice lungs. AB - Inhaled molecular oxygen has been widely used in humans to evaluate pulmonary ventilation using MRI. MR imaging has recently played a greater role in examining the morphologic and physiologic characteristics of mouse models of lung disease where structural changes are highly correlated to abnormalities in respiratory function. The motivation of this work is to develop oxygen-enhanced MR imaging for mice. Conventional human MR techniques cannot be directly applied to mouse imaging due to smaller dimensions and faster cardiac and respiratory physiology. This study examines the development of oxygen-enhanced MR as a noninvasive tool to assess regional ventilation in spontaneously breathing mice. An optimized cardiac-triggered, respiratory-gated fast spin-echo imaging sequence was developed to address demands of attaining adequate signal from the parenchyma, maintaining practical acquisition times, and compensating for rapid physiological motion. On average, a 20% T1-shortening effect was observed in mice breathing 100% oxygen as compared to air. The effect of ventilation was shown as a significant signal intensity increase of 11% to 16% in the mouse parenchyma with 100% oxygen inhalation. This work demonstrates that adequate contrast and resolution can be achieved using oxygen-enhanced MR to visualize ventilation, providing an effective technique to study ventilation defects in mice. PMID- 18421694 TI - Manganese-enhanced MRI detection of neurodegeneration in neonatal hypoxic ischemic cerebral injury. AB - In this study, we investigated the Mn-enhanced MRI (MEMRI) for detecting neurodegenerative processes in neonatal hypoxic-ischemic (H-I) cerebral injury. Seven-day-old rats were induced with H-I injury, and scanned for T1-weighted image (T1WI) and T2-weighted image (T2WI) with and without systemic MnCl2 administration. Serial histological analysis was performed for Mn-superoxide dismutase (Mn-SOD) and glutamine synthetase (GS), which are Mn-binding enzymes against the oxidative stress and glutamate excitotoxicity in neurodegeneration. In the acute phase (first 2 days), the ipsilateral lesion exhibited no Mn enhancement in T1WIs, with histology showing no Mn-SOD and GS production. In the mid-phase (from day 3), Mn enhancement was found in the cortex, basal ganglia, and hippocampus, correlating with local Mn-SOD and GS increase. In the late phase, the enhancement became more localized to the pericyst basal ganglia and cortex, and then gradually diminished. In T2WIs, a signal decrease was observed from day 3 in the corresponding regions. Hypointense voids gradually formed in the late phase, correlating with the local iron accumulation. H-I rats without Mn2+ administration exhibited similar but weak changes in T1WIs and T2WIs from days 14 and 7, respectively. These results indicate that Mn2+ may be a useful in vivo probe for monitoring Mn-SOD and GS enzymatic activities. PMID- 18421695 TI - Quantification of viable tumor microvascular characteristics by multispectral analysis. AB - Tumor heterogeneity complicates the quantification of tumor microvascular characteristics assessed by dynamic contrast-enhanced MRI (DCE-MRI). To address this issue a novel approach was developed that combines DCE-MRI with diffusion based multispectral (MS) analysis to quantify the microvascular characteristics of specific tumor tissue populations. Diffusion-based MS segmentation (feature space: apparent diffusion coefficient, T(2) and proton density) was performed to identify tumor tissue populations and the DCE-MRI characteristics were determined for each tissue class. The ability of this MS DCE-MRI technique to detect microvascular changes due to treatment with an antibody (G6-31) to vascular endothelial growth factor-A (VEGF) was evaluated in a tumor xenograft mouse model. Anti-VEGF treatment resulted in a significant reduction in K(trans) for the MS viable tumor tissue class (-0.0034 +/- 0.0022 min(-1), P < 0.01) at 24 hr posttreatment that differ significantly from the change observed in the control group (0.0002 +/- 0.0025 min(-1)). Viable tumor K(trans) for the anti-VEGF group was also reduced 62% relative to the pretreatment values (P < 0.01). Necrotic tissue classes were found to add only noise to DCE-MRI estimates. This approach provides a means to measure physiological parameters within the viable tumor and address the issue of tumor heterogeneity that complicates DCE-MRI analysis. PMID- 18421696 TI - Performance of a 200-MHz cryogenic RF probe designed for MRI and MRS of the murine brain. AB - A cryogenic 200-MHz RF transmit/receive probe (CryoProbe) made of copper and designed for murine brain studies is described. The probe operates at 30K and its performance was compared to a home-made surface coil of equal dimensions and a mouse head volume resonator, both operating at room temperature (RT). Since for small volumes of tissue interrogated (<5 mL) sample noise and thermal noise of the receiver are of comparable magnitude, considerable noise reduction can be achieved by cooling of the receiver system, e.g., using cold helium gas. The effect of temperature lowering on the signal-to-noise ratio (SNR) was assessed in phantom and in vivo murine brain studies using conventional MRI sequences (spin and gradient echo) and single voxel MR spectroscopy. Consistent sensitivity increases were observed for the CryoProbe. SNR increases by a factor>or=2 for the phantom experiments and by factors of 1.8 to 1.9 for in vivo experiments when compared to the RT surface coil for both MRI and MRS experiments. These results are in good agreement with estimated SNR gains for a circular surface coil probe cooled to 30K. The use of a cryogenic probe represents an economically attractive option to enhance the sensitivity in small animal MRI/MRS. PMID- 18421697 TI - High-resolution NMR imaging of paramagnetic liposomes targeted to a functionalized surface. AB - A major application of molecular MR imaging is receptor mapping of cells lining blood vessels with targeted contrast agents. Since these agents accumulate at interfaces, knowledge of their influence on the relaxation process in this specific configuration is a prerequisite for understanding their working principle. A methodology is presented to study the influence of targeted contrast agents on surface relaxation in vitro. Paramagnetic liposomes attached to a functionalized surface were studied with high-resolution NMR imaging. The surface was prepared by covering a solid substrate with a layer of collagen. Paramagnetic liposomes were targeted to this surface by functionalizing the liposomes with collagen adhesion protein CNA-35. With a saturation-recovery sequence, 1D magnetization profiles with a resolution of 5 microm were measured in water in contact with the surface. Analytical predictions, obtained with the Bloch-Torrey equation, perfectly agreed with the experimental data. Therefore, the magnitude of the surface relaxation rate could be determined from the measurements without any assumption. By using the relaxivity of liposome solutions the surface coverage by liposomes could be estimated. With the presented methodology the behavior of Gd-based targeted contrast agents at biological interfaces can be studied in vitro. Their influence on relaxation processes can be characterized and quantified. PMID- 18421698 TI - Chromosome instability in resection margins predicts recurrence of oral squamous cell carcinoma. PMID- 18421699 TI - Exploring the high-mass components of humic acid by laser desorption ionization mass spectrometry. AB - Leonardite and Elliot soil humic acids have been analyzed by laser desorption ionization mass spectrometry (LDI MS) in the m/z 4000-200,000 range. Positive ion mass spectra for each humic acid obtained under optimum conditions showed a broad high-mass distribution between m/z 20,000 and 80,000. The dependence of the mass distribution on instrumental parameters and solution conditions was used to investigate the nature of the high-mass peaks from humic acid spectra. Our data suggests that macromolecular ions and humic acid aggregates have the same probability of occurrence while cluster ion formation has a low probability of occurrence. PMID- 18421700 TI - Formation of metal-nicotianamine complexes as affected by pH, ligand exchange with citrate and metal exchange. A study by electrospray ionization time-of flight mass spectrometry. AB - Nicotianamine (NA) is considered as a key element in plant metal homeostasis. This non-proteinogenic amino acid has an optimal structure for chelation of metal ions, with six functional groups that allow octahedral coordination. The ability to chelate metals by NA is largely dependent on the pK of the resulting complex and the pH of the solution, with most metals being chelated at neutral or basic pH values. In silico calculations using pKa and pK values have predicted the occurrence of metal-NA complexes in plant fluids, but the use of soft ionization techniques (e.g. electrospray), together with high-resolution mass spectrometers (e.g. time-of-flight mass detector), can offer direct and metal-specific information on the speciation of NA in solution. We have used direct infusion electrospray ionization mass spectrometry (time-of-flight) ESI-MS(TOF) to study the complexation of Mn, Fe(II), Fe(III), Ni, Cu by NA. The pH dependence of the metal-NA complexes in ESI-MS was compared to that predicted in silico. Possible exchange reactions that may occur between Fe-NA and other metal micronutrients as Zn and Cu, as well as between Fe-NA and citrate, another possible Fe ligand candidate in plants, were studied at pH 5.5 and 7.5, values typical of the plant xylem and phloem saps. Metal-NA complexes were generally observed in the ESI-MS experiments at a pH value approximately 1-2 units lower than that predicted in silico, and this difference could be only partially explained by the estimated error, approximately 0.3 pH units, associated with measuring pH in organic solvent-containing solutions. Iron-NA complexes are less likely to participate in ligand- and metal-exchange reactions at pH 7.5 than at pH 5.5. Results support that NA may be the ligand chelating Fe at pH values usually found in phloem sap, whereas in the xylem sap NA is not likely to be involved in Fe transport, conversely to what occurs with other metals such as Cu and Ni. Some considerations that need to be addressed when studying metal complexes in plant compartments by ESI-MS are also discussed. PMID- 18421701 TI - Genetic polymorphisms for the study of multifactorial stroke. AB - Single-gene disorders explain only a minority of stroke cases. Stroke represents a complex trait, which is usually assumed to be polygenic. On this topic, the role of a wide number of candidate genes has been investigated in stroke through association studies, with controversial results. Therefore, it is difficult for the clinician to establish the validity and the level of clinical applicability of the previously reported associations between genetic factors and stroke. This review is an update and an extensive analysis of the more recent association studies conducted in stroke. We evaluated a number of studies on several candidate genes (including F5, F2, FGA/FGB/FGG, F7, F13A1, vWF, F12, SERPINE1, ITGB3/PLA1/PLA2/ITGA2B, ITGA2, GP1BA, ACE, AGT, NOS3, APOE, LPL, PON1, PDE4D, ALOX5AP, MTHFR, MTR, and CBS), providing a final panel of genes and molecular variants. We categorized this panel in relation to the degree of association with stroke, supported by the results of meta-analyses and case-control studies. Our findings could represent a useful tool to address further molecular investigations and to realize more detailed meta-analyses. PMID- 18421702 TI - Characterization of intermediate compounds formed upon photoinduced degradation of quinolones by high-performance liquid chromatography/high-resolution multiple stage mass spectrometry. AB - The paper deals with the photocatalytic transformation of two antibacterial agents, ofloxacin and ciprofloxacin, under simulated solar irradiation using titanium dioxide as photocatalyst. The investigation involved monitoring decomposition of the drugs, identifying intermediate compounds, assessing mineralization, and evaluating the toxicity of drug derivatives. High-resolution mass spectrometry was employed to assess evolution of the photocatalyzed process over time. Respectively 15 and 8 main species were identified after transformation of ofloxacin and ciprofloxacin. Through the full analysis of MS and MSn spectra and a comparison with parent drug fragmentation pathways, the different isomers were characterized. In the ofloxacin molecule, the initial transformation attacks are confined to the piperazine moiety and to the methyl groups, while the fluoroquinolone core is unmodified. Conversely, ciprofloxacin degradation involves two parts of the molecule: the piperazinic moiety and the quinolone moiety. All these intermediates are easily degraded and by 4 h mineralization is complete. Toxicity assays using Vibrio fischeri prove that neither ciprofloxacin nor its intermediates exhibit acute toxicity. In ofloxacin the secondary degradation products exhibit toxicity; a correlation exists between the evolution of the intermediate compounds and the toxicity connected to them. PMID- 18421703 TI - Protein kinase A RII-like (R2D2) proteins exhibit differential localization and AKAP interaction. AB - A-kinase anchoring proteins (AKAPs) bind to protein kinase A (PKA) via an amphipathic helix domain that interacts with a dimerization/docking domain on the regulatory (R) subunit of PKA. Four other mammalian proteins (ROPN1, ASP, SP17, and CABYR) also contain a highly conserved RII dimerization/docking (R2D2) domain, suggesting all four proteins may interact with all AKAPs in a manner similar to RII. All four of these proteins were originally detected in the flagellum of mammalian sperm. In this report, we demonstrate that all four R2D2 proteins are expressed in a wide variety of tissues and three of the proteins SP17, CABYR, and ASP are located in motile cilia of human bronchus and fallopian tubes. In addition, we detect SP17 in primary cilia. We also provide evidence that ROPN1 and ASP bind to a variety of AKAPs and this interaction can be disrupted with anchoring inhibitor peptides. The interaction of SP17 and CABYR with AKAPs appears to be much more limited. None of the R2D2 proteins appears to bind cAMP, a fundamental characteristic of the regulatory subunits of PKA. These observations suggest that R2D2 proteins utilize docking interactions with AKAPs to accomplish their function of regulating cilia and flagella. Based on location, affinity for AKAPs and lack of affinity for cAMP, it appears that each R2D2 protein has a unique role in this process. PMID- 18421704 TI - Origins of endomorphin-immunoreactive fibers and terminals in different columns of the periaqueductal gray in the rat. AB - Endomorphin 1 (EM1) and endomorphin 2 (EM2) are endogenous ligands for mu-opioid receptors (MOR). In the central nervous system, EM-immunoreactive (IR) neuronal cell bodies are located mainly in the hypothalamus and the nucleus tractus solitarius (NTS). EM-IR fibers and terminals are found widely distributed in many brain areas, including the different columns of the periaqueductal gray (PAG). The hypothalamus, NTS, and PAG are closely involved in modulation of vocalization, autonomic and neuroendocrine functions, pain, and defensive behavior through endogenous opioid peptides that bind to the MOR in these regions. Projections exist from both the hypothalamus and the NTS to the PAG. In order to examine whether there are EM1- and/or EM2-ergic projections from the hypothalamus and NTS to the PAG, immunofluorescence histochemistry for EM1 and/or EM2 was combined with fluorescent retrograde tracing. In rats that had Fluoro Gold (FG) injected into different columns of the PAG, some of the EM1- or EM2-IR neurons in the hypothalamus, but none in the NTS, were labeled retrogradely with FG. The majority of the EM1/FG and EM2/FG double-labeled neurons in the hypothalamus were distributed in the dorsomedial nucleus, areas between the dorsomedial and ventromedial nucleus, and arcuate nucleus; a few were also seen in the ventromedial, periventricular, and posterior nucleus. The present results indicate that the EM-IR fibers and terminals in the PAG originate principally from the hypothalamus. They also suggest that EMs released from hypothalamus-PAG projecting neurons might mediate or modulate various functions of the PAG through binding to the MOR. PMID- 18421705 TI - Sunlight ultraviolet irradiation and BRAF V600 mutagenesis in human melanoma. AB - The incidence of melanoma, the most lethal form of skin cancer, continues to increase in the Western world. In addition to genetic alterations in high- and low-susceptibility genes identified for melanoma, somatic mutations in BRAF gene occur frequently in human melanoma and are distinctively linked to sun exposure. Of significance is a single hotspot codon, i.e., BRAF V600, wherein up to 92% of all mutations arise. Recent work in our laboratory has demonstrated that solar ultraviolet (UV) irradiation triggers mutagenesis through induction of various DNA lesions, many of which capable of producing similar types of mutations, as those seen in BRAF V600 variants in human melanoma. In this review article, we have discussed application of "DNA damage-targeted mutagenicity" of solar UV irradiation for determining the mechanistic involvement of sunlight UV in BRAF V600 mutagenesis in human melanoma. We envision that establishing "DNA-damage derived mutagenesis" in this exceptionally unique target gene may resolve the underlying mechanism(s) of melanoma-genesis, thus helping define preventive and therapeutic measures against this malignant disease. PMID- 18421706 TI - Pigmented and nonpigmented ocelli in the brain vesicle of the ascidian larva. AB - The vertebrate-type opsin, Ci-opsin1, is localized in the outer segments of the photoreceptor cells of larvae of the ascidian Ciona intestinalis. The absorption spectrum of the photopigment reconstituted from Ci-opsin1 and 11-cis-retinal suggested that the photopigment is responsible for photic behavior of the larvae. The structure and function of Ci-opsin1-positive photoreceptor cells were examined by immunohistochemistry, confocal microscopy, electron microscopy, laser ablation, and behavioral analysis. Ciona larvae have three morphologically distinct groups of photoreceptor cells in the brain vesicle. Group I and group II photoreceptor cells are associated with the ocellus pigment cell on the right side of the brain vesicle. The outer segments of the group I photoreceptor cells are regularly arranged inside the small cavity encircled by the cup-shaped pigment cell. The outer segments of the group II photoreceptor cells are located outside the pigment cavity and exposed to the lumen of the brain vesicle. The outer segments of the group III photoreceptor cells are located near the otolith on the left ventral side of the brain vesicle. Thus, the brain vesicle of the ascidian larva has two ocelli: a 'conventional' pigmented ocellus containing the group I and group II photoreceptor cells and a novel nonpigmented ocellus solely consisting of the group III photoreceptor cells. Laser ablation experiments suggest that the pigmented ocellus is responsible for the photic swimming behavior. The nonpigmented ocellus might relate to later developmental or physiological events, such as metamorphosis, because Ci-opsin1 immunoreactivity appears in the late larval stage and becomes intense just before the onset of metamorphosis. PMID- 18421707 TI - The motor activity of mammalian axonemal dynein studied in situ on doublet microtubules. AB - Flagellar dynein generates forces that produce relative shearing between doublet microtubules in the axoneme; this drives propagated bending of flagella and cilia. To better understand dynein's role in coordinated flagellar and ciliary motion, we have developed an in situ assay in which polymerized single microtubules glide along doublet microtubules extruded from disintegrated bovine sperm flagella at a pH of 7.8. The exposed, active dynein remain attached to their respective doublet microtubules, allowing gliding of individual microtubules to be observed in an environment that allows direct control of chemical conditions. In the presence of ATP, translocation of microtubules by dynein exhibits Michaelis-Menten type kinetics, with V(max) = 4.7 +/- 0.2 microm/s and K(m) = 124 +/- 11 microM. The character of microtubule translocation is variable, including smooth gliding, stuttered motility, oscillations, buckling, complete dissociation from the doublet microtubule, and occasionally movements reversed from the physiologic direction. The gliding velocity is independent of the number of dynein motors present along the doublet microtubule, and shows no indication of increased activity due to ADP regulation. These results reveal fundamental properties underlying cooperative dynein activity in flagella, differences between mammalian and non-mammalian flagellar dynein, and establish the use of natural tracks of dynein arranged in situ on the doublet microtubules of bovine sperm as a system to explore the mechanics of the dynein microtubule interactions in mammalian flagella. PMID- 18421708 TI - Creatine monohydrate attenuates body fat accumulation in children with acute lymphoblastic leukemia during maintenance chemotherapy. AB - BACKGROUND: Corticosteroids are an important component of the treatment of acute lymphoblastic leukemia (ALL), with known significantly negative effects on bone and muscle. Creatine monohydrate (CrM) supplementation may be an adjunctive therapeutic strategy to attenuate some of these adverse effects. PROCEDURE: Nine children with ALL in the maintenance phase of treatment on the Dana-Farber Cancer Institute (DFCI) protocol 2000-2001 were treated with CrM (0.1 g/kg/day) for two sequential periods of 16 weeks (16 weeks treat > 6 weeks wash-out > 16 weeks treat). A cohort of children (N = 50) who were receiving the same chemotherapy at the same time served as natural history controls. Measurements included height, weight, body mass index (BMI), and lumbar spine bone mineral density (LS-BMD), whole body bone mineral content (WB-BMC), fat-free mass (FFM), and percent body fat (%BF) using dual-energy X-ray absorptiometry. RESULTS: Despite the long course of corticosteroid treatment for ALL, children showed significant increases in height, LS-BMD, WB-BMC and FFM over approximately 38 weeks (P < 0.05) during the study. There was an increase in BMI over time, but children taking CrM had a reduction, while the natural history group showed an increase in % BF (P < 0.05 for interaction). CONCLUSIONS: Children with ALL treated with corticosteroids as part of a maintenance protocol of chemotherapy showed an increase in % BF that was attenuated by CrM supplementation. PMID- 18421709 TI - C-erbB-2 expression and prognostic significance in osteosarcoma. AB - BACKGROUND: C-erbB-2 is postulated as a prognostic biological marker for osteosarcoma (OST). We investigated c-erbB-2 expression status in osteosarcomas and its prognostic significance. PROCEDURE: Archival pre-treatment tumor biopsies of 79 cases were stained by immunohistochemical (IHC) methods with CB11 antibody for cell-membrane specific c-erbB-2 expression. Relation with different variables was examined and survival rates were calculated. RESULTS: Median age was 13 years (7-17). Distant metastases were detected in 8 (10.1%) cases at initial diagnosis. 36/79 (45.6%) patients had positive c-erbB-2 staining by IHC. There was no significant difference between c-erbB-2 expression and presence of distant metastases at initial diagnosis or during follow-up, gender, age groups, tumor size, tumor sites, serum alkaline phosphatase, and lactate dehydrogenase levels. At a median follow-up of 67.4 months (9.2-246.1), 36 cases died, 20 were alive, and 23 abandoned treatment. For 58 cases without initial metastases who underwent definitive surgery, 5-year event-free (EFS) and overall survival (OS) rates were 50% and 62.1% in cases negative for c-erbB-2 expression, and 22.4% and 32.3% in cases positive for c-erbB-2 expression, respectively (EFS, P = 0.044; OS, P = 0.029). Five-year EFS and OS rates were significantly higher in cases with a tumor size <10 cm. Cox regression analyses revealed that only size persisted as a prognostic factor for EFS. C-erbB-2 expression status didn't appear as an independent prognostic factor. CONCLUSIONS: Overexpression of c-erbB-2 in OST was not correlated with metastatic potential. No independent prognostic significance was detected. The results do not support directed therapies for c-erbB-2 in OST. PMID- 18421710 TI - Fournier's gangrene and scrotal ulcerations during all-trans-retinoic acid therapy for acute promyelocytic leukemia. AB - Scrotal ulcers are a rare manifestation in patients with acute promyelocytic leukemia. Fournier's gangrene (FG) is even rarer. We describe three adolescents and young adults who developed scrotal ulcerations during induction with all trans-retinoic acid. One patient developed FG. These lesions are predominantly seen in Asian population. A good outcome with supportive management occurred in all the cases. PMID- 18421711 TI - Statistical analysis of personal radiofrequency electromagnetic field measurements with nondetects. AB - Exposimeters are increasingly applied in bioelectromagnetic research to determine personal radiofrequency electromagnetic field (RF-EMF) exposure. The main advantages of exposimeter measurements are their convenient handling for study participants and the large amount of personal exposure data, which can be obtained for several RF-EMF sources. However, the large proportion of measurements below the detection limit is a challenge for data analysis. With the robust ROS (regression on order statistics) method, summary statistics can be calculated by fitting an assumed distribution to the observed data. We used a preliminary sample of 109 weekly exposimeter measurements from the QUALIFEX study to compare summary statistics computed by robust ROS with a naive approach, where values below the detection limit were replaced by the value of the detection limit. For the total RF-EMF exposure, differences between the naive approach and the robust ROS were moderate for the 90th percentile and the arithmetic mean. However, exposure contributions from minor RF-EMF sources were considerably overestimated with the naive approach. This results in an underestimation of the exposure range in the population, which may bias the evaluation of potential exposure-response associations. We conclude from our analyses that summary statistics of exposimeter data calculated by robust ROS are more reliable and more informative than estimates based on a naive approach. Nevertheless, estimates of source-specific medians or even lower percentiles depend on the assumed data distribution and should be considered with caution. PMID- 18421712 TI - Effects of weak mobile phone - electromagnetic fields (GSM, UMTS) on event related potentials and cognitive functions. AB - Modern mobile phones emit electromagnetic fields (EMF) ranging from 900 to 2000 MHz which are suggested to have an influence on well-being, attention and neurological parameters in mobile phone users. Until now most studies have investigated Global System for Mobile Communications (GSM)-EMF and only very few studies have focused on Universal Mobile Telecommunications System (UMTS)-EMF. Therefore, we tested the effects of both types of unilaterally presented EMF, 1950 UMTS (0.1 and 1 W/kg) and pulsed 900 MHz GSM (1 W/kg), on visually evoked occipital P100, the P300 of a continuous performance test, auditory evoked central N100 and the P300 during an oddball task as well as on the respective behavioral parameters, reaction time and false reactions, in 15 healthy, right handed subjects. A double-blind, randomized, crossover application of the test procedure was used. Neither the UMTS- nor the GSM-EMF produced any significant changes in the measured parameters compared to sham exposure. The results do not give any evidence for a deleterious effect of the EMF on normal healthy mobile phone users. PMID- 18421713 TI - Biloma and pneumobilia in sickle cell disease. AB - Spontaneous biloma and pneumobilia developed in a patient with sickle cell disease (SCD) who presented with acute abdominal pain, jaundice, and fever. Bilomas are fluid-filled cysts that develop from a bile leak and are located outside of the biliary system. Pneumobilia is characterized by gas within the biliary system due to an abnormal connection between the biliary tree and adjacent organs, particularly the gastrointestinal tract. Although hepatobiliary complications are common in SCD, we report on biloma and pneumobilia in a young patient with this condition. PMID- 18421714 TI - 5,10-methylenetetrahydrofolate reductase (MTHFR) polymorphisms and the risk of acute lymphoblastic leukemia (ALL) in Filipino children. AB - BACKGROUND: 5,10-Methylenetetrahydrofolate reductase (MTHFR) is a critical enzyme in folate metabolism. Polymorphisms at the C677T and A1298C loci are associated with reduced activity; consequently more folate substrates are shunted toward thymidylate and DNA synthesis. Several studies have reported a reduced risk of developing ALL in children with MTHFR polymorphisms. The objective of this study was to determine the association between MTHFR polymorphisms and ALL in Filipino children. PROCEDURE: We conducted a case control study in children diagnosed with ALL at the Philippine General Hospital from 1/2001 through 12/2005. Bone marrow aspirate slides were reviewed by two expert hematologists to verify the morphologic diagnosis of ALL. DNA was isolated from the slides and MTHFR polymorphisms, C677T and A1298C, were determined using Taqman real-time PCR. Cord blood of healthy Filipino newborns served as control. RESULTS: There were a total of 191 ALL and 394 controls genotyped. The distribution of C677T polymorphisms was similar in the two groups (P = 1.0). However, for A1298C, there was significantly more AC and CC genotypes in the ALL compared to controls (P = 0.02; OR 1.57; CI: 1.08-2.28). The 1298C allele frequency for the control group was 36.8% and 677T allele frequency was 9.9%. CONCLUSION: A1298C polymorphisms is associated with an increased risk for ALL in Filipino children. This may be due to a difference in leukemia biology or to a high prevalence of folate deficiency in Filipinos. Our study reiterates the gene and environment interaction in leukemogenesis. PMID- 18421715 TI - Serum protein expression profiling in pediatric Hodgkin lymphoma: a report from the Children's Oncology Group. AB - BACKGROUND: The prognosis for children with Hodgkin lymphoma (HL) treated with a risk adjusted combination of radiation therapy and multi-drug chemotherapy has markedly improved. There remains a group of patients whose disease either recurs or does not respond to therapy. Protein expression profiling has been used to define protein characteristics of serum from adult patients in order to improve screening for early diagnosis. However, profiling for the purpose of staging and defining prognostic characteristics of childhood diseases is not well studied. The current stage-based risk assignment of HL cannot predict the patients within a risk group that are destined to recur or do not respond to therapy. Thus, a need exists to develop new methodologies to better stratify the risk classification of pediatric HL. PROCEDURE: We have completed a preliminary project to identify characteristic serum protein peaks determined by protein expression profiling in serum of 22 subjects with HL, 13 with stage II HL and 9 with stage III or IV. RESULTS: Protein profiling successfully discriminated between high grade (III/IV) HL and low grade (II) HL. CONCLUSION: These data lay the basis for prospective studies to identify protein expression profiles useful for diagnosis, prognosis, treatment stratification, and the follow-up of minimal residual disease. PMID- 18421716 TI - Neuropsychological outcome following intensity-modulated radiation therapy for pediatric medulloblastoma. AB - BACKGROUND: Combined cisplatin chemotherapy and cranial irradiation for treatment of medulloblastoma in children can cause significant ototoxicity and impair cognitive function and quality of life. We have previously demonstrated the conformal technique of intensity-modulated radiation therapy (IMRT) to reduce ototoxicity, however, it has been suggested that IMRT may increase risk of cognitive deficits compared to conventional radiation therapy (CRT). This study compared the impact of the two treatments on measures of neurocognitive functioning. PROCEDURE: Twenty-five pediatric patients with medulloblastoma were treated either with CRT or IMRT. In addition they received neurocognitive assessments to evaluate long-term functional outcome. Statistical analyses between the two groups were conducted to compare levels and profiles of performance on tests not confounded with hearing loss. RESULTS: When compared to CRT, children treated with IMRT did not perform more poorly on any of the measures. Both groups' mean performance was significantly lower than published norms on several of the measures employed. CONCLUSION: The benefit of reduced ototoxicity with IMRT does not appear to be at the cost of a decline in nonverbal intellectual abilities, visual-spatial skills, processing speed, or fine motor dexterity when compared to CRT in children with medulloblastoma. PMID- 18421717 TI - Familial infantile pyknocytosis in association with pulmonary hypertension. AB - Infantile pyknocytosis is a rare condition characterized by transient neonatal hemolytic anemia associated with increased pyknocyte count on blood smear. We describe three siblings with infantile pyknocytosis, born to consanguineous parents. The first and third siblings had neonatal hemolytic anemia that resolved spontaneously at 6 months. The second sibling presented at 11 days with severe hemolytic anemia along with pulmonary hypertension. He died at 39 days from sepsis. The findings support a possible autosomal recessive inheritance. We hypothesize that pulmonary hypertension may be secondary to or aggravated by neonatal hemolysis. PMID- 18421718 TI - Peripherally inserted central venous catheters are a good option for prolonged venous access in children with cancer. AB - BACKGROUND: A long term venous access device is essential in children with malignancies for the safe administration of medication and to avoid repeated painful venipunctures. The advantage of peripherally inserted central venous catheters (PICC) over conventional central venous catheter (CVC) is easy bedside insertion without need for general anesthesia and theatre time. The purpose of this study was to evaluate our experience with PICCs particularly with regard to catheter life, reason for removal and complications in children suffering from various malignancies. PROCEDURE: A retrospective analysis of all PICCs inserted in children with cancer was done with regard to the demographic data, catheter life, reason for removal, and complications. The latter two were evaluated in association with patient age, catheter days, and year of insertion. RESULTS: Of 127 catheters inserted in 127 children, median catheter life was 161 days with a total of 18,955 catheter days (for 124 patients, 3 lost to follow-up). Elective removal occurred in 63/101 (62.4%) PICCs and removal due to complications resulted in a complication rate of 2.41 per 1,000 catheter days. The common reasons for catheter removal were suspected infection, breakage/leakage, dislodgement, phlebitis, and occlusion with rates of 1.27, 0.57, 0.31, 0.06, and 0.06 per 1,000 catheter days, respectively. CONCLUSION: We found PICC to be a convenient, cheap, safe, and reliable device for long term intravenous access in children with malignancies. This was possible with the help of dedicated catheter care nurses. PMID- 18421719 TI - Recurrent idiopathic thrombocytopenic purpura in childhood. AB - BACKGROUND: Idiopathic thrombocytopenic purpura (ITP) is a common haematological disease during childhood, that usually has a benign course; however, literature on the recurrent form of the disease (rITP) is limited. PROCEDURE: rITP was characterized by intermittent episodes of thrombocytopenia (TP) followed by periods of recovery, unrelated to therapeutic intervention. We retrospectively reviewed features of patients with rITP, diagnosed and systematically followed up at our center, during the period 1975-2004. RESULTS: Forty-eight of 795 children with ITP (6.0 %) presented with rITP. The majority of patients (68.8%) had only one recurrence, whereas only one patient had four. A time interval between two episodes longer than 3 months (up to 96) was identified in 2/3 of episodes and <3 months in 1/3. The initial episode and the first recurrence mostly shared features of acute ITP; however, 22.9% of the episodes appeared with a chronic self-limited course. Bleeding manifestations were rare (18.6% of episodes) and mild, and they tended to occur in severely thrombocytopenic patients, mainly at the onset of the initial episode; intracranial hemorrhage (ICH) occurred in a toddler with short duration thrombocytopenia. Intravenous gamma globulin (IVIG) or corticosteroids were administered in 24.5% of episodes. None of the patients needed splenectomy. CONCLUSION: rITP is a rare, mild, self-limited type of ITP, although ICH may occur in a profoundly TP child. Recurrence may occur close or far apart to a previous isolated TP episode. The duration of episodes varies considerably from patient to patient and from episode to episode in the same patient. The pathogenesis of rITP still remains unclear. PMID- 18421720 TI - What do epidemiologists mean by 'population mixing'? AB - There is growing evidence that some chronic diseases are caused, or promoted, by infectious disease. 'Population mixing' has been used as a proxy for the range and dose of infectious agents circulating in a community. Given the speculation over the role of population mixing in many chronic diseases, we review the various methods used for measuring population mixing, and provide a classification of these. We recommend that authors fulfill two criteria in publications: measures are demonstrably associated with the putative risk factors for which population-mixing is acting as a proxy and fundamental characteristics of the chosen measures are clearly defined. PMID- 18421721 TI - Familial hemophagocytic lymphohistiocytosis in two brothers with X-linked agammaglobulinemia. AB - Hemophagocytic lymphohistiocytosis (HLH) is often familial and is associated with high mortality. Primary (familial) HLH is known to occur in children with mutations in perforin, Munc13-4, or syntaxin 11. We describe a case series of two brothers who developed HLH in the setting of X-linked agammaglobulinemia (XLA, Bruton's disease) and adenovirus infection. Further studies revealed absence of Bruton's tyrosine kinase (BTK) protein expression and a novel BTK mutation. PMID- 18421723 TI - Memory applications and electrical bistability of semiconducting nanoparticles: do the phenomena depend on bandgap? PMID- 18421722 TI - The management of patients with primary central nervous system (CNS) germinoma: current controversies requiring resolution. PMID- 18421724 TI - Electrogeneration of polypyrrole/alginate films for immobilization of glucose oxidase. AB - Electrogenerated PPy doped with pSA was used as a substrate for immobilization of GOD. This was achieved via covalent bonding of carboxyl groups of the main chain of alginate with amino groups of the enzyme. The pH-induced aggregation behavior of SA in aqueous solution was employed to provide optimum conditions for electrochemical preparation of PPy by galvanostatic methods. GOD was attached to the electrode surface by reaction between the carboxyl groups in the main chain of pSA with amino groups of GOD after treatment with EDC and NHS. The linkage of GOD enzyme to the conductive surface was characterized by ATR spectroscopy and SEM CV was used to demonstrate the bioactivity of the enzyme electrode toward glucose. PMID- 18421725 TI - Anthracene carboxyimides and their dimers. AB - Soluble anthracenedicarboxyimides have been prepared and undergo a photodimerisation of the anthracene skeleton, which is important for their application as antitumour agents, such as azonafides. Reaction under strongly alkaline conditions causes C-C coupling to form soluble dimeric fluorescent dyes with bathochromic absorption and fluorescence in the NIR region. These dyes are of special interest because of their absorption at longer wavelengths. PMID- 18421726 TI - Ruthenium-catalyzed synthesis of allylic alcohols: boronic acid as a hydroxide source. AB - Secondary allylic alcohols were synthesized from linear allylic halides or carbonates using a catalytic amount of a ruthenium complex in the presence of boronic acid. The effects of solvent, base, ruthenium precursor, and boronic acid were fully explored, and the scope of the reaction was extended to various substrates. We also describe a preliminary investigation towards an enantioselective process. PMID- 18421727 TI - Potential-energy and free-energy surfaces of glycyl-phenylalanyl-alanine (GFA) tripeptide: experiment and theory. AB - The free-energy surface (FES) of glycyl-phenylalanyl-alanine (GFA) tripeptide was explored by molecular dynamics (MD) simulations in combination with high-level correlated ab initio quantum chemical calculations and metadynamics. Both the MD and metadynamics employed the tight-binding DFT-D method instead of the AMBER force field, which yielded inaccurate results. We classified the minima localised in the FESs as follows: a) the backbone-conformational arrangement; and b) the existence of a COOH...OC intramolecular H-bond (families CO(2)H(free) and CO(2)H(bonded)). Comparison with experimental results showed that the most stable minima in the FES correspond to the experimentally observed structures. Remarkably, however, we did not observe experimentally the CO(2)H(bonded) family (also predicted by metadynamics), although its stability is comparable to that of the CO(2)H(free) structures. This fact was explained by the former's short excited-state lifetime. We also carried out ab initio calculations using DFT-D and the M06-2X functional. The importance of the dispersion energy in stabilising peptide conformers is well reflected by our pioneer analysis using the DFT-SAPT method to explore the nature of the backbone/side-chain interactions. PMID- 18421728 TI - Porphyrin dimers bridged by an electrochemically switchable unit. PMID- 18421729 TI - Growth of larger hydrocarbons in the ionosphere of titan. PMID- 18421730 TI - Direct allylation of aldimines catalyzed by C2-symmetric N,N'-dioxide-Sc(III) complexes: highly enantioselective synthesis of homoallylic amines. PMID- 18421731 TI - Non-innocence of N-heterocyclic carbene ligands: intermolecular C-H activation in allyl palladium NHC complexes. PMID- 18421732 TI - Pd(II) and Pt(II) complexes with amphiphilic ligands: formation of micelles and [5]rotaxanes with alpha-cyclodextrin in aqueous solution. AB - [3]Pseudorotaxanes [1(alpha-CD)(2)][X] (X=Cl, NO(3)), prepared from reaction of an N-alkylbipyridinium [4,4'-bpy-N-(CH(2))(10)OC(6)H(3)-3,5-(OMe)(2)][X] ([1][X]) and alpha-CD, react with M(NO(3))(2)(en) (M=Pd, Pt; en=1,2-ethylenediamine) in a 2:1 molar ratio to afford [5]rotaxanes [M{(4,4'-bpy-N-(CH(2))(10)OC(6)H(3)-3,5 (OMe)(2))(alpha-CD)(2)}(2) (en)][NO(3)](4) ([2(alpha-CD)(4)][NO(3)](4), M=Pd; [3(alpha-CD)(4)][NO(3)](4), M=Pt). A similar reaction of [1][Cl] with [M(NO(3))(2)(en)] (M=Pd, Pt) produces amphiphilic Pd and Pt complexes, [2][NO(3)](4) and [3][NO(3)](4). Complexes [2][NO(3)](4) and [3][NO(3)](4) form micelles in the presence of small amounts of dyes (Nile red and pyrene) in water. The critical micelle concentration (CMC) was determined by the absorption peak of the dye, which is encapsulated in the micelles in solution. Micelle formation is confirmed by dynamic light scattering measurement of the solution and TEM (transmission electron microscopy) images of the micelles deposited from the solution. Addition of alpha-CD to the aqueous solution containing these amphiphilic complexes results in degradation of the micelle structure and the formation of [5]rotaxanes, [2(alpha-CD)(4)][NO(3)](4) and [3(alpha CD)(4)][NO(3)](4). PMID- 18421733 TI - Asymmetric organocatalysis: from infancy to adolescence. AB - After an initial period of validating asymmetric organocatalysis by using a wide range of important model reactions that constitute the essential tools of organic synthesis, the time has now been reached when organocatalysis can be used to address specific issues and solve pending problems of stereochemical relevance. This Review deals with selected studies reported in 2006 and the first half of 2007, and is intended to highlight four main aspects that may be taken as testimony of the present status and prospective of organocatalysis: a) chemical efficiency; b) discovery of new substrate combinations to give new asymmetric syntheses; c) development of new catalysts for specific purposes by using mechanistic findings; and d) applications of organocatalytic reactions in the asymmetric total synthesis of target natural products and known compounds of biological and pharmaceutical relevance. PMID- 18421734 TI - Fluorous tags catching on microarrays. PMID- 18421735 TI - Janus colloids formed by biphasic grafting at a Pickering emulsion interface. PMID- 18421736 TI - Enantioselective activation of aldehydes by chiral phosphoric acid catalysts in an aza-ene-type reaction between glyoxylate and enecarbamate. PMID- 18421737 TI - Structural complexity in intermetallic alloys: long-periodic order beyond 10 nm in the system BaSn3/BaBi3. PMID- 18421738 TI - Schiff base macrocycles containing pyrroles and pyrazoles. AB - A double nucleophilic substitution reaction of 3,5-bis(chloromethyl)pyrazole with pyrroles generates a novel pyrrole-pyrazole hybrid building block, the pyrazole analogue to tripyrrane. Vilsmeier-Haack formylation produces the corresponding dialdehyde, which was used in the formation of a series of nonaromatic Schiff base macrocycles. NMR and UV/Vis spectroscopy and single-crystal diffractometry were used to characterize the novel macrocycles. The solid-state structures of select free bases and protonated members of this class of macrocycles display a range of intra- and intermolecular hydrogen-bonding patterns that suggest their use in molecular-recognition systems. They also contain an acid-sensitive chromophore. Their acid-base and anion-recognition properties were ascertained; alas, only modest anion-selective spectroscopic signatures could be detected by using UV/Vis and (1)H NMR spectroscopy. The macrocycles proved resistant toward oxidation to their aromatic congeners. The pyrrole-pyrazole building blocks presented are potentially useful for the synthesis of a range of pyrazole analogues of all-pyrrole macrocycles. PMID- 18421739 TI - High-resolution separation of peptides by sodium dodecyl sulfate-polyacrylamide gel "focusing". AB - As a follow-up of our previous report (Anal. Chem. 2007, 79, 821-827) on analytical SDS-PAGE focusing, a refinement of the method for separation of peptides in the small to medium M(r) range (0.5-10 kDa) is here reported, based on a shallow gradient of immobilized positive charges (0-10 mM) onto a minimally sieving polyacrylamide gel matrix (4%T, 2.5%C). Unlike conventional SDS-PAGE, which rarely can achieve the separations of polypeptide chains below a critical value of 10 kDa, the present method can be fine-tuned to perform such separations even down to a size of only 500 Da. In the case of larger fragments, the major peptide zones are shown, under microscope observation, to be composed by envelopes of bands as narrow as 20-100 microm, spaced at regular intervals of 100 150 microm. It is hypothesized that such larger peptides could form complexes with rather small SDS micelles and that such peptide-SDS complexes could differ in charge by just a single negative charge. PMID- 18421742 TI - Celebrating the 80th birthday of professor Stellan Hjerten. PMID- 18421743 TI - The role of cyclodextrins in chiral capillary electrophoresis. AB - The members of the enantiomeric pairs frequently show rather different biological effects, so their chiral selective synthesis, pharmacological studies and analysis are necessary. CE has unique advantages in chiral analysis. The most frequently used chiral selectors are CDs in this field. This paper gives a short view on the advantages on CE in direct chiral separations, emphasizing the role of CDs. The reason for the broad selectivity spectra of CDs is discussed in detail. The physical background of chiral selective separations is briefly shown in CE. Their interaction mechanisms are shortly defined. The general trend of their use is statistically evaluated. Most frequently used CDs and CD derivatives are characterized. Advantages of ionizable CDs and single-isomer derivatives are shown. The general trend of their use is established. PMID- 18421744 TI - A highly stereoselective hydrogen-bond-mediated Michael-Michael cascade process through dynamic kinetic resolution. PMID- 18421746 TI - Bioactive constituents of myrrh and frankincense, two simultaneously prescribed gum resins in chinese traditional medicine. PMID- 18421747 TI - Cobinamides as structural probes in B12-biosynthesis: synthesis and spectroscopic analysis of isomers of Co alpha,Co beta-dicyanocobinamide. AB - Vitamin B(12) and its coenzyme forms are cobalamins (i.e., cobamides, 'complete' with a 5,6-dimethylbenzimidazole nucleotide base), in which the particular corrinoid moiety of the cobinamides is conjugated to alpha-ribazole-3'-phosphate via a phosphate-diester group. Aside of being provided with their particular reactivity, required for their functions as organometallic cofactors in B(12) dependent enzymes, the cobalamins also depend upon their specific three dimensional buildup, to be able to adapt the unique constitution of 'base-on' corrinoids by intramolecular Co-coordination of the nucleotide base. We report rational partial syntheses and detailed spectral analyses of three close cobinamide isomers in their Co(alpha),Co(beta)-dicyano forms: of 13-epicobinamide (also called neocobinamide), of 176(S)-epicobinamide, and of 176-isocobinamide. Neocobinamide was obtained under acidic conditions as a degradation product of vitamin B(12). 176(S)-Epicobinamide and 176-isocobinamide were prepared by condensation of cobyric acid with (2S)-1-aminopropan-2-ol and with 3-aminopropan 1-ol, respectively. Natural cobinamide represents the corrinoid nucleus produced by proper microbial biosynthesis (as intermediate for the further assembly of the 'complete' corrinoid cofactors) or is required in some microorganisms, such as Escherichia coli, as an exogenously supplied unit for further biosynthetic buildup. The three compounds may thus be of use as structural probes for the biosynthetic capacity and tolerance in microorganisms, and (some of them) may serve as substrates as well, for further biosynthetic 'completion' of corrinoid cofactors or their analogues. PMID- 18421748 TI - Cytotoxic triterpenoids from the root bark of Helicteres angustifolia. AB - Three new triterpenoids, 3beta-acetoxy-27-[(E)-cinnamoyloxy]lup-20(29)-en-28-oic acid methyl ester (1), 3beta-acetoxy-27-[(4-hydroxybenzoyl)oxy]lup-20(29)-en-28 oic acid (2), and 3beta-acetoxy-27-[(4-hydroxybenzoyl)oxy]olean-12-en-28-oic acid methyl ester (3), together with nine known triterpenoids, 4-12, were isolated from the root bark of Helicteres angustifolia. The structures of these compounds were established on the basis of spectroscopic methods including 2D-NMR experiments. All twelve compounds were tested for their cytotoxic activities against human colorectal cancer (COLO 205), human hepatoma (Hep G2), and human gastric cancer (AGS) cell lines in vitro. Among them, compounds 2, 3, 3beta-O [(E)-coumaroyl]betulinic acid (6), and pyracrenic acid (7) showed significant cytotoxic activities against human cancer cells COLO 205 and AGS. PMID- 18421749 TI - Interaction of isoquinoline alkaloid palmatine with deoxyribonucleic acids: binding heterogeneity, and conformational and thermodynamic aspects. AB - The binding heterogeneity, conformational aspects, and energetics of the interaction of the cytotoxic plant alkaloid palmatine have been studied with various natural and synthetic DNAs. The alkaloid binds to calf thymus and Escherichia coli DNA that have mixed AT and GC sequences in almost equal proportions with positive cooperativity, while, with Clostridium perfringens and Micrococcus lysodeikticus DNA with predominantly high AT and GC sequences, respectively, noncooperative binding was observed. On further investigation with synthetic DNAs, the binding was observed to be cooperative with polymers like poly(dA).poly(dT) and poly(dG).poly(dC) having poly(purine)poly(pyrimidine) sequences, while with polymers poly(dA-dT).poly(dA-dT), poly(dA-dC).poly(dG-dT) and poly(dG-dC).poly(dG-dC), which have alternating purine-pyrimidine sequences, a non-cooperative binding phenomenon was observed. This suggests the binding heterogeneity of palmatine to the two types of sequences of base pairs. Circular dichroism (CD) studies revealed that the binding induced conformational changes in all the DNAs, but more importantly, the bound alkaloid molecules acquired induced optical activity, and the extent was dependent on the AT content and showed AT base-pair specificity. Energetics of the interaction of the alkaloid studied by highly sensitive isothermal titration calorimetry revealed that the binding was in most cases exothermic and favored by both enthalpy and entropy changes, while, in the case of the homo and hetero AT polymers, the same was predominantly entropy-driven. This study defines base-pair-dependent heterogeneity, conformational aspects, and energetics of palmatine binding to DNA. PMID- 18421750 TI - New hepatoprotective coumarinolignoids from Mallotus apelta. AB - Three new coumarinolignoids, malloapelins A-C (1-3, resp.), together with three known coumarinolignoids, cleomiscosin A (4), cleomiscosin B (5), and 5' demethylaquillochin (6), were isolated from the roots of Mallotus apelta MUELL. ARG. Compounds 1-6 are three pairs of regioisomeric coumarinolignoids. Their structures were elucidated on the basis of spectral evidence. Compounds 3 showed promising hepatoprotective activity against D-galactosamine-induced toxicity in WB-F344 rat hepatic epithelial stem-like cells. PMID- 18421751 TI - New stemona alkaloids from the roots of Stemona sessilifolia. AB - From the roots of Stemona sessilifolia, three new stemona-type alkaloids, namely stemosessifoine (1), isooxymaistemonine (2), and isomaistemonine (3), along with eight known alkaloids (bisdehydrostemoninine, isobisdehydrostemoninine, tuberostemonine, bisdehydrotuberostemonine, bisdehydrostemoninine, isobisdehydrostemoninine, stemoninine, and protostemonine), were isolated. Their structures were determined on the basis of extensive 2D-NMR spectroscopic-data analysis and by comparison with reported values in the literature. Compound 1 is a structurally unprecedented alkaloid, and it is depicted to be bioconverted from tuberostemonine as the precursor. Isooxymaistemonine (2) showed a positive effect on the human high-density lipoprotein (HDL) receptor gene CD36 and LIMP II analogous-1 (CLA-1) at the dosage of 10 microg/ml. PMID- 18421752 TI - Determination of trace alkaline phosphatase by solid-substrate room-temperature phosphorimetry based on Triticum vulgare lectin labeled with fullerenol. AB - Fullerenol (F) shows a strong and stable room-temperature phosphorescence (RTP) signal on the surface of nitrocellulose membrane (NCM) at lambda ex max/ lambda em max =542.0/709.4 nm. When modified by dodecylbenzenesulfonic acid sodium salt (DBS), fullerenol emits a stronger signal. It was also found that quantitative specific affinity-adsorption reaction can be carried out between Triticum vulgare lectin (WGA) labeled with DBS-F and alkaline phosphatase (ALP) on the surface of NCM, and the product obtained (WGA-ALP-WGA-F-DBS) emits a strong and stable RTP signal. Furthermore, the content of ALP was proportional to the DeltaI(p) value. Based on the facts above, a new method for the determination of trace amounts of ALP by affinity-adsorption solid-substrate room-temperature phosphorimetry (AA-SS RTP) was established, using fullerenol modified with DBS to label WGA. The detection limit was 0.011 fg spot(-1) (corresponding concentration: 2.8x10(-14) g ml(-1), namely 2.8x10(-16) mol l(-1)). This method with high sensitivity, accuracy, and precision has been successfully applied to the determination of the content of ALP in human serum survey and forecast human disease, and the results are tallied with those using alkaline phosphatase kits. The mechanism for the determination of ALP using AA-SS-RTP was also discussed. PMID- 18421753 TI - With the great complexity unveiling, can we still decipher the interaction between gut flora and the host in inflammatory bowel disease to find out the mechanism and cause? How? PMID- 18421754 TI - Mass measurement accuracy comparisons between a double-focusing magnetic sector and a time-of-flight mass analyzer. AB - We report a direct comparison of the mass measurement accuracies (MMAs) obtained on different mass spectrometry instrument types; a magnetic sector as the 'gold standard' and an electrospray ionization time-of-flight (ESI-TOF) instrument. Sixty samples, obtained from the Department of Chemistry at North Carolina State University, were analyzed on each instrument. Data are presented and compared between the different instruments. The average absolute MMAs achieved for the magnetic sector and Agilent ESI-TOF mass spectrometers were 3.0 and 1.1 ppm, respectively. PMID- 18421755 TI - Carboxamides combining favorable olfactory properties with insect repellency. AB - By a simple methodology, we have synthesized 68 carboxamides, with structural analogy to the most efficient mosquitoes repellent DEET (N,N-diethyl-m-toluamide, 9). Eight of them have very promising olfactory profiles. In addition, 34 carboxamides have been tested for their repellency properties against three breeds of cockroach; six of them are excellent in this domain. Two of these, the 2-ethyl-N-isopropyl-N-methylbutanamide and N,N-diallyl-2-ethylbutanamide combine both the positive aspects, and are thus very good candidates for use in functional perfumery, and more particularly in home-care formulations. PMID- 18421756 TI - Synthesis of resveratrol analogues, and evaluation of their cytotoxic and xanthine oxidase inhibitory activities. AB - Thirteen resveratrol (=5-[(E)-2-(4-hydroxyphenyl)ethenyl]benzene-1,3-diol) analogues with a CHO group have been prepared by partial synthesis from resveratrol. The synthesized compounds have been evaluated for their cytotoxic activity against a human nasopharyngeal epidermoid tumor cell line KB, as well as for their xanthine oxidase inhibitory activity. Compounds 2, 3, and 6a showed the most significant cytotoxic activities against the cell line KB, and compound 2 also exhibited strong xanthine oxidase inhibitory activity. PMID- 18421757 TI - Towards new antimalarial drugs: synthesis of non-hydrolyzable phosphate mimics as feed for a predictive QSAR study on 1-deoxy-D-xylulose-5-phosphate reductoisomerase inhibitors. AB - The conversion of 1-deoxy-D-xylulose-5-phosphate (DOXP) to 2-C-methyl-D erythritol-4-phosphate (MEP) is effectively blocked by 1-deoxy-D-xylulose-5 phosphate reductoisomerase (Dxr) inhibitors such as the natural antibiotic fosmidomycin. Prediction of binding affinities for closely related Dxr ligands as well as estimation of the affinities of structurally more distinct inhibitors within this class of non-hydrolyzable phosphate mimics relies on the synthesis of fosmidomycin derivatives with a broad range of target affinity. Maintaining the phosphonic acid moiety, linear modifications of the lead structure were carried out in an effort to expand the SAR of this physicochemically challenging class of compounds. Synthetic access to a set of phosphonic acids with inhibitory activity (IC(50)) in the range from 1 to >30 microM vs. E. coli Dxr and 0.4 to 20 microM against P. falciparum Dxr is reported. PMID- 18421758 TI - Synthesis of novel pyrrolo[3,4-d]pyrazole-dicarboxylic acids and evaluation of their interaction with glutamate receptors. AB - Chiral pyrazoline amino acids (3aR,4S,6aR)-1a and (3aR,4S,6aR)-1b, and (3aS,6S,6aS)-2a and (3aS,6S,6aS)-2b, which are conformationally constrained analogues of glutamic and homoglutamic acid, respectively, were prepared via a strategy based on the 1,3-dipolar cycloaddition of a nitrile imine to methyl N Boc-3,4-didehydro-(S)-prolinate. The new 'amino acids' were tested for activity at ionotropic glutamate receptors. Solely the derivative (3aR,4S,6aR)-1a, which is structurally related to the previously described 4,5-dihydroisoxazole analogue (S)-CIP-A, turned out to be a potent and selective agonist for the AMPA receptors. The biological activity is due to the interaction with the orthosteric glutamate binding site. PMID- 18421759 TI - N-glucosyl-1H-indole derivatives from Cortinarius brunneus (Basidiomycetes). AB - Two new N-glucosylated indole alkaloids were isolated from fruiting bodies of the basidiomycete Cortinarius brunneus (Pers.) Fr. The structures were elucidated by means of the spectroscopic data. Additionally, the very recently reported compounds N-1-beta-glucopyranosyl-3-(carboxymethyl)-1H-indole (3) and N-1-beta glucopyranosyl-3-(2-methoxy-2-oxoethyl)-1H-indole (4) could be detected. Compound 3 is the N-glucoside of the plant-growth regulator 1H-indole-3-acetic acid (IAA), but, in contrast, it does not exhibit auxin-like activity in an Arabidopsis thaliana tap root elongation assay. PMID- 18421760 TI - Abnormal crosstalk between pancreatic acini and macrophages during the clearance of apoptotic cells in chronic pancreatitis. AB - In chronic pancreatitis (CP), both the progressive loss of acinar parenchyma and aggressive fibro-inflammatory reactions ultimately lead to irreversible organ destruction. Dying cells are normally removed by macrophages and elimination is associated with anti-inflammatory cytokine switch. We investigated whether defective clearance of damaged acini by macrophages such as compromised phagocytosis or altered cytokine reaction occurs in CP and thus represents a causative link between acinar loss and fibro-inflammation. In a checkerboard-like co-culture system, we assessed normal and CP macrophages for their phagocytic and cytokine responses to dying pancreatic acinar cells of normal or CP origin by FACS, confocal microscopy, QRT-PCR, and ELISA. In CP, phagocytosis of apoptotic acini by macrophages was not impaired; however, the associated cytokine responses were gradually perturbed. Most interestingly, only normal acini suppressed TGFbeta1 expression and accumulation specifically in normal macrophage cultures, while CP acini lost this ability. Both types of apoptotic acini induced pro inflammatory cytokine bursts of varying strength in both types of macrophages; however, the most significant difference (more than 50-fold higher expression of IL-1beta, IL-6, and IL-8) was evident between CP/CP and normal/normal combinations, indicating that acinar and macrophage alterations synergistically lead to the ultimate CP-specific bias. In combination with in situ data comparing circulating inflammatory cells to pancreatic resident ones, our results indicate that cytokine expression in inflammatory cells undergoes spatiotemporal modulation, most likely through a successive interplay of acinar, stromal, and circulating factors. Thus, clearance of injured pancreatic acini by macrophages is associated with a unique cytokine reaction which may constitute a basis for progression of SAPE (sentinel acute pancreatitis event) to the irreversible fibro inflammation in CP. PMID- 18421761 TI - Phagocyte dysfunction and inflammatory bowel disease. AB - Inflammatory bowel diseases are common chronic inflammatory disorders. The majority are idiopathic and can be broadly divided into Crohn's disease and ulcerative colitis. Their cause is unknown, but most hypotheses focus on a primary role for T-cell dysfunction. Conversely, there is a collection of congenital disorders of phagocyte function that result not only in immunodeficiency but also in noninfectious inflammatory bowel disease. In all cases, the latter is strikingly reminiscent of the clinical and pathological features of Crohn's disease. This coincides with recent work demonstrating that despite previous emphasis on adaptive immune dysfunction, patients with Crohn's disease actually possess an unusually weak acute innate inflammatory response. This review consolidates the literature on inflammatory bowel disease in congenital immunodeficiencies and considers the role of phagocyte dysfunction in Crohn's disease. Concepts about pathogenesis and treatment that can be carried across these disorders are also discussed. PMID- 18421762 TI - Role of probiotic therapy in IBD. AB - There is mounting evidence that probiotic therapy may alter disease expression in both animal models of IBD and in patients with IBD. The effects appear to be modest at best and may reflect the choice of probiotic organism, the variability in concentrations of organisms administered, and the variability of the diseases being treated. This review examines the data of all fully published articles currently available for the role of probiotics in the treatment of IBD. PMID- 18421763 TI - Matrix-assisted laser desorption/ionisation mass spectrometry imaging of lipids in rat brain tissue with integrated unsupervised and supervised multivariant statistical analysis. AB - To date matrix-assisted laser desorption/ionisation mass spectrometry imaging (MALDI-MSI) analysis has been largely concerned with mapping the distribution of known analytes in tissues. An important step in the progression of its applications is the determination of unknown variants for metabolite and protein profiling in both clinical studies and studies of disease. Principal component analysis (PCA) is a statistical approach which can be used as a means of determining latent variables in multivariate data sets. In the work reported here, PCA, in both unsupervised and supervised modes, has been used to differentiate brain regions based on their lipid composition determined by MALDI MSI. PCA has been shown to be useful in the determination of hidden variables between spectra taken from six regions of brain tissue. It is possible to identify ions of interest from the loadings plot which are likely to be more prominent in the different regions of the brain and thus differentiating between white and grey matter. It is also possible to distinguish between the grey Cerebellar Cortex and the Hippocampal formation, due to the grey Cerebellar Cortex having a positive PC2 and the Hippocampal formation having a negative PC2 score; this is only possible in supervised PCA with this data set because with unsupervised PCA the two regions overlap. PMID- 18421764 TI - Leukocytapheresis in a girl with severe ulcerative colitis refractory to corticosteroids, infliximab, and cyclosporine A. PMID- 18421765 TI - Mass spectra of tetrahydroisoquinoline-fused 1,3,2-O,N,P- and 1,2,3-O,S,N heterocycles: influence of ring size and fusion, of present heteroatoms, substituent effects and of the stereochemistry on fragmentation. AB - The electron ionization (EI) mass spectra of a variety of stereoisomeric tricyclic 1,3,2-oxazaphosphino[4,3-a]isoquinolines (1-4), 1,2,3-oxathiazino[4,3 a]isoquinoline-4-oxides (5-7) and the -4,4-dioxides (8-10) of oxazaphospholo- and oxathiazolo[4,3-a]- (11, 12, 15 and 16) and -[3,4-b]isoquinolines (13, 14 and 17) were recorded. Ring size and fusion, the different heteroatoms (P and S) and substituents on the ring systems strongly influence the mass spectra. In addition, mass spectra of the stereoisomers of compounds 1, 2 and 13, 14 revealed stereochemically relevant differences which are not observed for the other pairs of isomers. PMID- 18421766 TI - Computational view of surface based organic mass spectrometry. AB - Surface based mass spectrometric approaches fill an important niche in the mass analysis portfolio of tools. The particular niche depends on both the underlying physics and chemistry of molecule ejection as well as experimental characteristics. In this article, we use molecular dynamics computer simulations to elucidate the fundamental processes giving rise to ejection of organic molecules in atomic and cluster secondary ion mass spectrometry (SIMS), massive cluster impact (MCI) mass spectrometry, and matrix-assisted laser desorption ionization (MALDI) mass spectrometry. This review is aimed at graduate students and experimental researchers. PMID- 18421767 TI - Proteomics in gram-negative bacterial outer membrane vesicles. AB - Gram-negative bacteria constitutively secrete outer membrane vesicles (OMVs) into the extracellular milieu. Recent research in this area has revealed that OMVs may act as intercellular communicasomes in polyspecies communities by enhancing bacterial survival and pathogenesis in hosts. However, the mechanisms of vesicle formation and the pathophysiological roles of OMVs have not been clearly defined. While it is obvious that mass spectrometry-based proteomics offers great opportunities for improving our knowledge of bacterial OMVs, limited proteomic data are available for OMVs. The present review aims to give an overview of the previous biochemical, biological, and proteomic studies in the emerging field of bacterial OMVs, and to give future directions for high-throughput and comparative proteomic studies of OMVs that originate from diverse Gram-negative bacteria under various environmental conditions. This article will hopefully stimulate further efforts to construct a comprehensive proteome database of bacterial OMVs that will help us not only to elucidate the biogenesis and functions of OMVs but also to develop diagnostic tools, vaccines, and antibiotics effective against pathogenic bacteria. PMID- 18421768 TI - Mass spectrometric analysis of illicit drugs in wastewater and surface water. AB - Residues of illicit drugs have been recently found in urban wastewater and surface water. Their levels reflect the amount of drugs collectively excreted by consumers and can therefore be used to estimate drug abuse. An overview of the most widely used illicit drugs and of the analytical methods used for their detection in wastewater and surface water is presented here. Solid-phase extraction and high performance liquid chromatography-tandem mass spectrometry are the techniques that have been used for these investigations. Instrumental conditions and fragmentation patterns of illicit drugs and their metabolites are described. PMID- 18421769 TI - Male reproductive skew, paternal relatedness, and female social relationships. AB - Female social relationships among primates are thought to be shaped by socio ecological factors and phylogenetic constraints. We suggest that patterns of paternal relatedness among females influence measures of social tolerance that have been used to classify species into different social relationship categories. As kin support and kin preference have only been measured for matrilineal kin and related individuals exchange less aggression and have a higher conciliatory tendency, the observed low nepotism levels and high tolerance levels may be an artifact of hidden paternal relatedness among the nonkin category. Using comparative data on macaques, we investigate this hypothesis using male reproductive skew as a proxy for paternal relatedness. Within the limitations of the study we show that populations classified as being less nepotistic, and more tolerant exhibit higher levels of reproductive skew. This first result and the reasoning behind may motivate future students of social relationships to take paternal relatedness into consideration. Potential implications of this finding if repeated with larger samples include that variation in aspects of macaque social relationships may be explained without considering phylogeny or the strength of between-group contest competition for food. PMID- 18421770 TI - Impact of ecologically relevant heat shocks on Hsp developmental function in the vetigastropod Haliotis asinina. AB - Heat shock proteins (Hsps) are essential for cellular maintenance, normal differentiation and morphogenesis, and protection against a range of environmental stresses. It is unknown which of these roles takes precedence when they are required simultaneously. Here we examined the impact of thermal stress on the complex developmental expression patterns of HasHsp70 and HasHsp90A in the vetigastropod Haliotis asinina. We find that near-lethal heat shocks do not alter the spatial demarcation of Hsp expression despite such treatments impacting on the external character of the embryos. Using a suite of molecular markers that are both coexpressed with the Hsps (i.e. in ventrolateral ectoderm and prototroch) and expressed in tissues that have lower (basal) Hsp expression (e.g. serotonergic nervous system and shell gland), we determined that Hsp-expressing tissues do not incur markedly less thermal damage than adjacent tissues. To explore the relationship of Hsp expression with sensitivity of specific cell territories to heat shock, we focused on the formation of the prototroch, a tissue where HasHsp70 and HasHsp90A are coexpressed. By heat shocking at specific developmental stages, we determined that the most sensitive period of prototroch development is during its early specification and differentiation, which overlaps with the time the Hsps are expressed at their highest levels in these cells. This correlation is consistent with heat shock impairing the function of Hsps in regions of the H. asinina embryo undergoing morphogenesis. PMID- 18421771 TI - Structure-function studies of glutamate synthases: a class of self-regulated iron sulfur flavoenzymes essential for nitrogen assimilation. AB - Glutamate synthases play with glutamine synthetase an essential role in nitrogen assimilation processes in microorganisms, plants, and lower animals by catalyzing the net synthesis of one molecule of L-glutamate from L-glutamine and 2 oxoglutarate. They exhibit a modular architecture with a common subunit or region, which is responsible for the L-glutamine-dependent glutamate synthesis from 2-oxoglutarate. Here, a PurF- (Type II- or Ntn-) type amidotransferase domain is coupled to the synthase domain, a (beta/alpha)8 barrel containing FMN and one [3Fe-4S]0,+1 cluster, through a approximately 30 angstroms-long intramolecular tunnel for the transfer of ammonia between the sites. In bacterial and eukaryotic GltS, reducing equivalents are provided by reduced pyridine nucleotides thanks to the stable association with a second subunit or region, which acts as a FAD-dependent NAD(P)H oxidoreductase and is responsible for the formation of the two low potential [4Fe-4S]+1,+2 clusters of the enzyme. In photosynthetic cells, reduced ferredoxin is the physiological reductant. This review focus on the mechanism of cross-activation of the synthase and glutaminase reactions in response to the bound substrates and the redox state of the enzyme cofactors, as well as on recent information on the structure of the alphabeta protomer of the NADPH-dependent enzyme, which sheds light on the intramolecular electron transfer pathway between the flavin cofactors. PMID- 18421772 TI - From anchorage dependent proliferation to survival: lessons from redox signalling. AB - Anchorage to extracellular matrix (ECM) is essential for the execution of the mitotic program of nontransformed cells as they need simultaneous signals starting from mitogenic molecules, as growth factors (GFs), and adhesive agents belonging to ECM. Reactive oxygen species play a key function during both GF and integrin receptor signalling and are therefore recognised to have a synergistic function with several others transducers for anchorage-dependent growth (ADG). Indeed, redox-regulated proteins include protein tyrosine phosphatases, protein tyrosine kinases, small GTPases, cytoskeleton proteins, as well as several transcription factors. In this review, we focus on the role of reactive oxygen species (ROS) as key second messengers granting a proper executed mitosis for anchorage-dependent cells through redox regulation of several downstream targets. Besides, redox signals elicited by ECM contact assure a protection from anoikis, a specific apoptosis induced by lack of anchorage. Cancer cells frequently show a deregulation of ROS production and a constitutive oxidative stress has been associated to the achievement of an invasive phenotype. Hence, in cancer cells, the constitutive deregulation of both mitogenic and survival pathways, likely mimicking autocrine/adhesive signals, helps to guide the transformed cells to escape the innate apoptotic response to abolish the signals started by cell/ECM contact, thus sustaining the spreading of anchorage-independent cancer cells and the metastases growth. PMID- 18421773 TI - Brain mitochondrial dysfunction in aging. AB - Aging of mammalian brain is associated with a continuous decrease of the capacity to produce ATP by oxidative phosphorylation. The impairment of mitochondrial function is mainly due to diminished electron transfer by complexes I and IV, whereas inner membrane H+ impermeability and F1-ATP synthase activity are only slightly affected. Dysfunctional mitochondria in aged rodents show decreased rates of respiration and of electron transfer, decreased membrane potential, increased content of the oxidation products of phospholipids and proteins, and increased size and fragility. In aging mice, the activities of brain mitochondrial enzymes (complexes I and IV and mtNOS) are linearly correlated with neurological performance (tightrope and T-maze tests) and with median life span and negatively correlated with the mitochondrial content of lipid and protein oxidation products. Conditions that increased mice median life span, such as moderate exercise, vitamin E supplementation, caloric restriction, and high spontaneous neurological activity; also improved neurological performance and mitochondrial function in aged brain. The diffusion of mitochondrial NO and H2O2 to the cytosol is decreased in the aged brain and may be a factor for reduced mitochondrial biogenesis. PMID- 18421774 TI - Mechanisms of aging and energy metabolism in Caenorhabditis elegans. AB - Aging studies on diverse species ranging from yeast to man have culminated in the delineation of several signaling pathways that influence the process of senescent decline and aging. While understanding these interlinked signal-transduction cascades is becoming even more detailed and comprehensive, the cellular and biochemical processes they impinge upon to modulate the rate of senescent decline and aging have lagged considerably behind. This fundamental question is one of the most important challenges of modern aging research and has been the focus of recent research efforts. Emerging findings provide insight into the facets of cellular metabolism which can be fine-tuned by upstream signaling events to ultimately promote longevity. Here, we survey the mechanisms regulating aging in the simple nematode worm Caenorhabditis elegans, aiming to highlight recent discoveries that shed light into the interface between aging signaling pathways and cellular energy metabolism. Our objective is to review the current understanding of the processes involved and discuss mechanisms that are likely conserved in higher organisms. PMID- 18421775 TI - Carnitine palmitoyltransferase 1: central to cell function. PMID- 18421776 TI - How does Nutrigenomics impact human health? PMID- 18421777 TI - IUBMB and the Mediterranean. PMID- 18421778 TI - Biochemistry and molecular biology in Greece: history and perspectives. PMID- 18421779 TI - Resveratrol: one molecule, many targets. AB - Resveratrol is one of the numerous polyphenolic compounds found in several vegetal sources. In recent years, the interest in this molecule has increased exponentially following the major findings that resveratrol (i) is shown to be chemopreventive in some cancer models, (ii) is cardioprotective, and (iii) has positive effects on several aspects of metabolism, leading to increased lifespan in all the metazoan models tested thus far, including small mammals. Such remarkable properties have elicited a vast interest towards the identification of target proteins of resveratrol and have led to the identification of enzymes inhibited by resveratrol and others whose activation is enhanced. In the vast majority of cases, resveratrol displays inhibitory/activatory effects in the micromolar range, which is potentially attainable pharmacologically, although targets with affinities in the nanomolar range have also been reported. Here, we provide an overview of the various classes of enzymes known to be inhibited (or activated) by resveratrol. It appears that resveratrol, as a pharmacological agent, has a wide spectrum of targets. The biological activities of resveratrol may thus be dependent on its simultaneous activity on multiple molecular targets. PMID- 18421781 TI - Biochemistry and molecular biology research achievements in Greece. PMID- 18421780 TI - Heparin--a unique stimulator of human colon cancer cells' growth. AB - The cancer microenvironment and the interactions between cancer and surrounding tissue cells are thought to play a pivotal role in tumor development and progression. Glycosaminoglycans (GAGs)/proteoglycans (PGs) are major constituents of the extracellular matrix, the composition of which may affect various cellular functions. In the present study, the effects of GAGs on the proliferation of HT29, SW1116, and HCT116 human colon cancer cell lines were examined using exogenously added GAGs, an inhibitor of endogenous GAG sulfation and specific glycosidase digestions. Our results demonstrate that colon cancer cell growth was exclusively stimulated by exogenously added heparin and insensitive to endogenous GAGs/PGs production, in a sulfation pattern-related manner. Treatment of the tested cell lines with the FGF-2 neutralizing antibody showed that the stimulatory effect of heparin on the cells' growth was not FGF-2-dependent. Responsiveness of colon cancer cell lines to exogenous heparin/heparan sulfate may play a role in their growth and metastasis. PMID- 18421782 TI - The history of biochemistry in Turkey and the Turkish Biochemical Society. PMID- 18421783 TI - Development of biochemistry and molecular biology in Malta. PMID- 18421784 TI - The Mediterranean Institute for Life Sciences--an attempt at scientific excellence in Croatia. PMID- 18421785 TI - Biochemistry and molecular biology in Portugal: an overview of past and current contributions. PMID- 18421786 TI - The Spanish Society of Biochemistry and Molecular Biology, the development of biochemistry in Spain, and IUBMB. PMID- 18421787 TI - Addressing PAH biodegradation in Greece: biochemical and molecular approaches. PMID- 18421788 TI - Bridging the research gap between the Northern and Southern shores of the Mediterranean. PMID- 18421789 TI - Biochemistry and molecular biology in Morocco: overview of scientific research and education. PMID- 18421790 TI - The 4-1BB ligand and 4-1BB expressed on osteoclast precursors enhance RANKL induced osteoclastogenesis via bi-directional signaling. AB - The 4-1BB is a costimulatory molecule similar to the receptor activator of NF kappaB ligand (RANKL), both of which are key factors for the differentiation of osteoclasts and are expressed mainly by activated T cells. The 4-1BB shares common signaling pathways with RANK, suggesting a potential role in osteoclastogenesis. In this study, the role of 4-1BB and 4-1BB ligand (4-1BBL) in osteoclastogenesis was investigated using 4-1BB(-/-) and 4-1BB(+/+) mice. Osteoclast precursors normally express 4-1BB and 4-1BBL after exposure to RANKL, which was confirmed by semi-quantitative RT-PCR and flow cytometry. The 4-1BB(-/- )mice had a slightly increased bone mass accompanied by a reduced osteoclastogenic ability of 4-1BB(-/-) bone marrow-derived macrophages (BMM) ex vivo. In addition, 4-1BB(-/-) BMM demonstrated hypophosphorylation of JNK and p38 and decreased induction of c-Fos in response to RANKL stimulation. Retroviral transduction of wild-type as well as partial-length 4-1BB, which lacks TNF receptor-associated factor 2-binding sites for signaling, restored the osteoclastogenic ability of 4-1BB(-/-) BMM. Furthermore, both recombinant 4-1BB and 4-1BBL enhanced RANKL-induced osteoclastogenesis by 4-1BB(+/+) BMM and the induction of c-Fos and NFATc1.Together, these results indicate that 4-1BBL and 4 1BB expressed on osteoclast precursors enhance RANKL-induced osteoclastogenesis via bi-directional signaling, findings that may delineate the complex nature of the 4-1BBL and 4-1BB interaction. PMID- 18421791 TI - Airway epithelial cells modify immune responses by inducing an anti-inflammatory microenvironment. AB - The upper airways are prone to contact with pathogenic as well as non-pathogenic microbes, therefore immune recognition principles have to be tightly controlled. Here we show that human BEAS-2B bronchial epithelial cells inhibited secretion of the pro-inflammatory cytokines TNF-alpha and IL-12 by monocytes, macrophages and dendritic cells. This inhibitory effect could be transferred by supernatant of resting BEAS-2B cells and was also observed when primary murine tracheal epithelial cells were prepared. In contrast to inhibition of pro-inflammatory cytokine secretion epithelial cell-conditioned dendritic cells showed increased expression of IL-10 and arginase-1, thus displaying properties of alternative activation. Accordingly, Toll-like receptor-mediated up-regulation of CD40, CD86 and PD-L2 (CD273) on murine dendritic cells was reduced in the presence of bronchial epithelial cell supernatant. However, expression of negative regulatory PD-L1 (CD274) was increased and dendritic cell induced proliferation of T lymphocytes was diminished. Epithelial cells also showed a direct inhibitory effect on T lymphocyte proliferation and this was due to the constitutive secretion of TGF-beta by bronchial epithelial cells. Moreover, epithelial cell conditioned T lymphocytes showed increased differentiation towards IL-10 producing Tr1 cells. The results indicate that bronchial epithelial cells induce a non-inflammatory microenvironment that regulates local immune homeostasis. PMID- 18421792 TI - Suppression of viral replication with highly active antiretroviral therapy has no impact on the functional profile of HIV-specific CD8(+) T cells. AB - A better control of viral replication in long-term non-progressors has been associated with polyfunctional CD8(+) T cell responses. However, low levels of HIV replication could be the cause rather than the consequence of enhanced immune responses in long-term non-progressors. The functional profile and the expansion ability of HIV-Gag- and HIV-Nef-specific CD8 responses were analysed measuring the production of MIP-1beta, IL-2, TNF-alpha and expression of CD107, using polychromatic flow cytometry, in 36 HIV-infected patients at baseline and after 12 months of highly active antiretroviral therapy (HAART) and complete viral suppression. Most patients presented detectable Gag and Nef responses both at baseline and after 1 year of HAART, with a significant decline after achieving viral suppression. At baseline, the majority of CD8(+) response was due to cells producing only MIP-1beta or simultaneously MIP-1beta and CD107. The functional profile did not significantly change after achieving complete viral suppression with HAART. Therefore, control of HIV-1 replication after 1 year of HAART had no significant impact on the quality of HIV-1-specific CD8 response, but the effects of treatment in long-term, or of early HAART are not known. Thus, it is still uncertain whether multifunctional CD8 responses are the cause or consequence of low plasma viremia. PMID- 18421794 TI - Executive function deficits in short-term abstinent cannabis users. AB - OBJECTIVES: Few cognitive tasks are adequately sensitive to show the small decrements in performance in abstinent chronic cannabis users. In this series of three experiments we set out to demonstrate a variety of tasks that are sufficiently sensitive to show differences in visual memory, verbal memory, everyday memory and executive function between controls and cannabis users. METHODS: A series of three studies explored cognitive function deficits in cannabis users (phonemic verbal fluency, visual recognition and immediate and delayed recall, and prospective memory) in short-term abstinent cannabis users. Participants were selected using snowball sampling, with cannabis users being compared to a standard control group and a tobacco-use control group. RESULTS: The cannabis users, compared to both control groups, had deficits on verbal fluency, visual recognition, delayed visual recall, and short- and long-interval prospective memory. There were no differences for immediate visual recall. CONCLUSIONS: These findings suggest that cannabis use leads to impaired executive function. Further research needs to explore the longer term impact of cannabis use. PMID- 18421793 TI - B7-H1 restricts neuroantigen-specific T cell responses and confines inflammatory CNS damage: implications for the lesion pathogenesis of multiple sclerosis. AB - The co-inhibitory B7-homologue 1 (B7-H1/PD-L1) influences adaptive immune responses and has been proposed to contribute to the mechanisms maintaining peripheral tolerance and limiting inflammatory damage in parenchymal organs. To understand the B7-H1/PD1 pathway in CNS inflammation, we analyzed adaptive immune responses in myelin oligodendrocyte glycoprotein (MOG)(35-55)-induced EAE and assessed the expression of B7-H1 in human CNS tissue. B7-H1(-/-) mice exhibited an accelerated disease onset and significantly exacerbated EAE severity, although absence of B7-H1 had no influence on MOG antibody production. Peripheral MOG specific IFN-gamma/IL-17 T cell responses occurred earlier and enhanced in B7-H1( /-) mice, but ceased more rapidly. In the CNS, however, significantly higher numbers of activated neuroantigen-specific T cells persisted during all stages of EAE. Experiments showing a direct inhibitory role of APC-derived B7-H1 on the activation of MOG-specific effector cells support the assumption that parenchymal B7-H1 is pivotal for delineating T cell fate in the target organ. Compatible with this concept, our data investigating human brain tissue specimens show a strong up-regulation of B7-H1 in lesions of multiple sclerosis. Our findings demonstrate the critical importance of B7-H1 as an immune-inhibitory molecule capable of down regulating T cell responses thus contributing to the confinement of immunopathological damage. PMID- 18421795 TI - Accuracy of classification of breast ultrasound findings based on criteria used for BI-RADS. AB - OBJECTIVE: To assess the accuracy of categorization of breast ultrasound findings based on scoring for malignancy using the sonographic breast imaging-reporting and data system (BI-RADS). METHODS: Breast ultrasound was performed in 2462 patients between 2001 and 2004 at our unit. Sonographic findings were scored using analog criteria as in BI-RADS for breast ultrasound (mass shape, margin, orientation, posterior acoustic features, lesion boundary, echo pattern). Each lesion was described using these features and classified into categories 1 to 5 according to the BI-RADS for breast ultrasound. Categorization and biopsy results were compared. RESULTS: In twenty-two (0.9%) patients breast ultrasound could not be evaluated because of extreme density of tissue. Normal breast ultrasound belonging to Category 1 was found in 871 (35.4%) patients. Simple cysts classified as Category 2 were observed in 712 (28.9%) women. In 491 (19.9%) patients, apparently benign solid masses (Category 3) were found. Suspicious masses were observed in 225 (9.1%) women and masses highly suggestive of malignancy were found in 141 (5.7%) patients (Categories 4 and 5, respectively). Histological examinations were available from 84 (17.1%) masses that had been classified by BI-RADS as Category 3, in 97 (43.1%) from Category 4 and 106 (75.2%) from Category 5. Accordingly, the rate of malignant findings was 1.2% (n = 1) in Category 3, 17% (n = 16) in Category 4 and 94% (n = 100) in Category 5. CONCLUSION: Scoring breast ultrasound findings for malignancy based on criteria used for BI-RADS breast ultrasound has a high accuracy, comparable to that obtained by BI-RADS for mammography. PMID- 18421796 TI - The bioconversion of mountain pine beetle-killed lodgepole pine to fuel ethanol using the organosolv process. AB - Lodgepole pine (Pinus contorta) killed by mountain pine beetle (Dendroctonus ponderosae) (BLP) was compared with healthy lodgepole pine (HLP) for bioconversion to ethanol and high-value co-products. The BLP and HLP chips were pretreated using an ethanol organosolv process at a variety of severities. It was shown that the BLP was easier to pretreat and delignify than were the HLP chips. The resulting pretreated BLP substrate had a lower residual lignin, lower degree of polymerization of cellulose, lower cellulose crystallinity, smaller fiber size and thereby a better enzymatic hydrolysability than did the HLP substrates. However, under the same conditions, the BLP showed lower substrate yield and cellulose recovery than did the HLP, which likely resulted from the excessive hydrolysis and subsequent decomposition of the cellulose and hemicellulose during the pretreatment. The BLP wood yielded more ethanol organosolv lignin than was obtained with the HLP material. The HLP lignin had a lower molecular weight and narrower distribution than did the BLP lignin. It appears that the beetle killed LP is more receptive to organosolv pretreatment other than a slightly lower recovery of carbohydrates. PMID- 18421797 TI - The roles of galactitol, galactose-1-phosphate, and phosphoglucomutase in galactose-induced toxicity in Saccharomyces cerevisiae. AB - The uptake and catabolism of galactose by the yeast Saccharomyces cerevisiae is much lower than for glucose and fructose, and in applications of this yeast for utilization of complex substrates that contain galactose, for example, lignocellulose and raffinose, this causes prolonged fermentations. Galactose is metabolized via the Leloir pathway, and besides the industrial interest in improving the flux through this pathway it is also of medical relevance to study the Leloir pathway. Thus, genetic disorders in the genes encoding galactose-1 phosphate uridylyltransferase or galactokinase result in galactose toxicity both in patients with galactosemia and in yeast. In order to elucidate galactose related toxicity, which may explain the low uptake and catabolic rates of S. cerevisiae, we have studied the physiological characteristics and intracellular metabolite profiles of recombinant S. cerevisiae strains with improved or impaired growth on galactose. Aerobic batch cultivations on galactose of strains with different combinations of overexpression of the genes GAL1, GAL2, GAL7, and GAL10, which encode proteins that together convert extracellular galactose into glucose-1-phosphate, revealed a decrease in the maximum specific growth rate when compared to the reference strain. The hypothesized toxic intermediate galactose-1 phosphate cannot be the sole cause of galactose related toxicity, but indications were found that galactose-1-phosphate might cause a negative effect through inhibition of phosphoglucomutase. Furthermore, we show that galactitol is formed in S. cerevisiae, and that the combination of elevated intracellular galactitol concentration, and the ratio between galactose-1-phosphate concentration and phosphoglucomutase activity seems to be important for galactose related toxicity causing decreased growth rates. PMID- 18421798 TI - Magnetic cell separation using nano-sized bacterial magnetic particles with reconstructed magnetosome membrane. AB - Magnetic nanoparticles produced by magnetotactic bacterium, bacterial magnetic particles (BacMPs), covered with a lipid bilayer membrane (magnetosome membrane) can be used to separate specific target cells from heterogeneous mixtures because they are easily manipulated by magnets and it is easy to display functional proteins on their surface via genetic engineering. Despite possessing unique and valuable characteristics, the potential toxicity of BacMPs to the separated cells has not been characterized in detail. Here, a novel technique was developed for the reconstruction of magnetosome membrane of BacMPs expressing protein A (protein A-BacMPs) to reduce cytotoxicity and the newly developed nanomaterial was then used for magnetic cell separation. The development of the magnetosome membrane-reconstructed protein A-BacMP was based on the characteristics of the Mms13 anchor protein, which strongly binds to the magnetite surface of BacMPs. Treatment of protein A-BacMPs with detergents removed contaminating proteins but did not affect retention of Mms13-protein A fusion proteins. The particle surfaces were then reconstructed with phosphatidylcholine. The protein A-BacMPs containing reconstructed magnetosome membranes remained dispersible and retained the ability to immobilize antibody. In addition, they contained few membrane surface proteins and endotoxins, which were observed on non-treated protein A BacMPs. Magnetic separation of monocytes and B-lymphocytes from the peripheral blood was achieved with high purity using magnetosome membrane-reconstructed protein A-BacMPs. PMID- 18421799 TI - The membrane bioreactor: a novel tool to grow anammox bacteria as free cells. AB - In a membrane bioreactor (MBR), fast growth of anammox bacteria was achieved with a sludge residence time (SRT) of 12 days. This relatively short SRT resulted in a -for anammox bacteria--unprecedented purity of the enrichment of 97.6%. The absence of a selective pressure for settling, and dedicated cultivation conditions led to growth in suspension as free cells and the complete absence of flocs or granules. Fast growth, low levels of calcium and magnesium, and possibly the presence of yeast extract and a low shear stress are critical for the obtainment of a completely suspended culture consisting of free anammox cells. During cultivation, a population shift was observed from Candidatus "Brocadia" to Candidatus "Kuenenia stuttgartiensis." It is hypothesized that the reason for this shift is the higher affinity for nitrite of "Kuenenia." The production of anammox bacteria in suspension with high purity and productivity makes the MBR a promising tool for the cultivation and study of anammox bacteria. PMID- 18421800 TI - The loudness dependence of the auditory evoked potential (LDAEP) as an in vivo biomarker of central serotonergic function in humans: rationale, evaluation and review of findings. AB - OBJECTIVES: The loudness dependence of the auditory evoked potential (LDAEP) has been proposed as a valid means of non-invasively assessing in vivo central serotonin (5-hydroxytryptamine, 5-HT) function in humans. The specificity and sensitivity of the LDAEP to changes in 5-HT neurotransmission have recently been explored directly in a number of pharmacological and genetic studies. Subsequently, this review was undertaken in an attempt to critically evaluate the potential role of the LDAEP as a marker of the central 5-HT function. DESIGN: Findings from clinical, experimental animal and human studies examining the relationship between the LDAEP and the 5-HT system as well as other neurochemical systems including dopaminergic, glutamatergic and the cholinergic systems were reviewed. RESULTS: The majority of evidence for an association between the LDAEP and 5-HT has come from animal studies. Indirect studies in clinical disorders of presumed serotonergic dysfunction have been circumstantial and inconsistent with more recent investigations utilising direct genetic association studies also providing conflicting reports. Pharmacological studies in humans provide overwhelming evidence that the LDAEP is insensitive to acute changes in 5-HT function, with additional evidence outlining sensitivity to other neurotransmitter systems including the glutamatergic system. CONCLUSIONS: The available evidence suggests that the LDAEP lacks sensitivity and specificity to acute changes in serotonergic neurotransmission. Overall the findings do not provide strong support for its utility as a marker of central 5-HT function. However the LDAEP shows more promise as a potential predictor of antidepressant treatment response and this predictive ability may provide the basis for future research involving the LDAEP. PMID- 18421801 TI - The use of effect sizes to characterize the nature of cognitive change in psychopharmacological studies: an example with scopolamine. AB - Drug induced cognitive change is generally investigated using small sample sizes. In terms of null hypothesis significance testing (NHST) this can render a meaningful change non-significant, as a result of insufficient power in the statistical model. NHST leads to 'all or none' thinking, where a non-significant result is interpreted as an absence of change. An effect size calculation indicates the magnitude of change which has occurred post-intervention, and therefore whether a significant result is meaningful. We used a scopolamine challenge to demonstrate the usefulness of effect sizes. The aim of the study was to determine how effect sizes could describe the cognitive changes that occur following administration of subcutaneous scopolamine (s.c. scopolamine). Twenty four healthy young males (M = 32.6, sd = 4.5 years) were administered placebo and 0.2 mg, 0.4 mg & 0.6 mg of s.c. scopolamine using a 4-way crossover design. Memory, learning, psychomotor function, attention and executive function were assessed. Scopolamine significantly impaired performance on all tasks in a dose and time related manner. These results demonstrate the functionality of change scores to draw comparisons between different times and doses. This methodology overcomes the limitations of comparisons between studies using different tasks, doses and time at which cognitive functions are measured. PMID- 18421803 TI - Resistance monitoring in codling moth: a need for standardization. AB - BACKGROUND: A wide range of methods has been used for monitoring resistance in Cydia pomonella L. However, the results from specific tests are not always concordant and they generate doubts over the extent to which the data represent the field situation. Therefore, the variability in the expression of resistance mechanisms and in resistance ratios between developmental stages and their reliability in a field situation were studied. RESULTS: Neonate larvae showed the highest insecticide resistance, except to spinosad. Resistance ratios were from 2 to 9.5, depending on the insecticide and the strain tested, when compared with the susceptible strain. Spinosad exhibited the highest resistance ratio for diapausing larvae (4.4, 12.2 and 4.4 for Raz, Rdfb and RDelta strains, respectively). Enzymatic ratios were higher in adults than in neonates or diapausing larvae, and the highest enzymatic ratios were observed for mixed function oxidases (7.3-36.7 for adults of resistant strains). CONCLUSIONS: The sensitivity of different instars depends on the insecticides used, and there is a differential expression of resistance mechanisms between stages. For the population considered in this study, tests on neonates provided the most consistent results to the field situation, but adult response to chlorpyrifos ethyl and azinphos-methyl in the laboratory was also consistent with the field results. PMID- 18421802 TI - Diminished 5-HT functioning in CBCL pediatric bipolar disorder-profiled ADHD patients versus normal ADHD: susceptibility to rapid tryptophan depletion influences reaction time performance. AB - OBJECTIVE: There is a current debate on characterizing children with pediatric bipolar disorder (PBD) through a profile within the child behaviour checklist (CBCL), and on the involvement of the 5-HT system in the underlying neurobiological processes of PBD. The aim of the present paper was to investigate reaction time performance in patients with CBCL-PBD and to discriminate ADHD from ADHD with CBCL-PBD with respect to diminished 5-HT functioning and reaction time. METHODS: Twenty-two patients with ADHD received the rapid tryptophan depletion test (RTD) thus lowering the central-nervous 5-HT synthesis rate within a placebo controlled double-blind within-subject crossover design. Reaction time was assessed using a competitive reaction time game with low and high provocation after both depletion and placebo intake. The study sample was divided into high and low scorers according to their CBCL-PBD scores. RESULTS: Comparing those six patients with the highest and clinically significant CBCL-PBD scores versus those six patients with the lowest, patients with a high CBCL-PBD score showed a slower reaction time under RTD compared to patients with low CBCL-PBD scores after high provocation. CBCL-'aggression' discriminated between the two groups. CONCLUSIONS: The results suggest alterations in 5-HT functioning in CBCL-PBD-spectrum patients, and 'aggression' as a potential moderator variable to ADHD. PMID- 18421804 TI - Why can't I drink grapefruit juice if I take a statin? PMID- 18421805 TI - Neurobehavioral Teratology Society membership list (as of 1/15/2008). PMID- 18421806 TI - Power lines. PMID- 18421808 TI - An interview with Nicholas Swindale. PMID- 18421807 TI - Not in our backyard. PMID- 18421809 TI - [Patients with mental retardation in a psychiatric hospital]. AB - OBJECTIVE: Because only few data are available, inpatient care of patients with mental retardation was evaluated to get a valid data base concerning quality and quantity of care and to reveal their deficits. METHODS: All patients with first admission to a psychiatric hospital between 1996 and 2002 were included in te cohort study (n=9727). The analyses were based on the psychiatric basis documentation system (DGPPN-BADO). 192 patients (2%) had the diagnosis of a mental retardation according to ICD-10. RESULTS: 62% of patients suffered from a mild (F70), 26% from a moderate (F71) and 9% from a severe mental retardation (F72). The most frequent comorbid disorders were disruptive behaviour disorders, adjustment disorders, alcohol-related disorders, Schizophrenia and mood disorders. Mean length of stay was 34.8 days, rate of rehospitalisation within one year was 0.3. GAF improved on 11.8, CGI (improvement scale) was 3.4. Some predictors for these outcome criteria could be revealed. CONCLUSIONS: The data allow a depth insight into inpatient care of patients with mental retardation. They deliver important clues for quality improvement in psychiatric hospital and for collaboration with outpatient services. PMID- 18421810 TI - Rodeo doctor. PMID- 18421811 TI - No smoke without fire? The cannabis debate. PMID- 18421812 TI - Reefer madness? PMID- 18421813 TI - All together now. PMID- 18421814 TI - Life peers. Interview by Catherine Jackson. PMID- 18421815 TI - Partnerships work. Interview by Harriet Pearce-Willis. PMID- 18421816 TI - Spreading the word. Interview by Catherine Jackson. PMID- 18421817 TI - No more wasted lives. Interview by Catherine Jackson. PMID- 18421818 TI - The bigger picture. If racism exists in society, then surely it must influence mental health services. PMID- 18421819 TI - Welfare writes. The DWP is sometimes right, sometimes wrong, and sometimes downright cheeky. PMID- 18421821 TI - Sustaining a recovery-orientated workforce. PMID- 18421822 TI - You don't know like I know. PMID- 18421823 TI - Psychosocial interventions during crisis resolution. PMID- 18421824 TI - One foot in the door. PMID- 18421825 TI - Recovery and beyond. PMID- 18421826 TI - Network. Centre for Advocacy in Mental Health in India. PMID- 18421827 TI - View point. Mental health stereotypes in defence of mental health services. PMID- 18421828 TI - Two new staining procedures for quantitative estimation of proteins on electrophoretic strips. AB - Two new procedures are described for the estimation of protein by direct photometry on electrophoretic strips. The protein complexes of procion brilliant blue RS and coomassie brilliant blue R250 are shown to follow Beer's law up to 50 and 20 microg/cm, respectively. The lower limits of detection are 2 and 0.5 microg/cm. Within these ranges the absolute amount of protein can be estimated within a single test with an accuracy of about +/-10%. The major contribution to the error arises from uneven application of the samples. Relative concentrations within a mixture of proteins can be evaluated to an accuracy better than +/-3%. Technical details of the procedures and of the equipment required are given in full, and their areas of usefulness discussed. PMID- 18421829 TI - Symposium: thalamocortical relationships. Introductory remarks. PMID- 18421830 TI - Organization of the mammalian thalamus and its relationships to the cerebral cortex. AB - The available data concordantly suggest that the mammalian thalamus consists of three divisions different from each other in their phylogenetic and ontogenetic development and in their relations to the cortex. The epithalamus (paraventricular complex, habenular complex, and the pretectal group of nuclei) is entirely independent of the endbrain in all mammals and undergoes a strong reduction in higher forms. The dorsal thalamus is entirely dependent on the endbrain. Each nucleus of this division has a restricted projection upon the endbrain without which it cannot survive. The dorsal thalamic nuclei are classified as extrinsic or intrinsic depending on whether or not they receive a substantial portion of their afferents from extra-thalamic sources. It can be shown that the neocortex of primitive mammals consists largely of projection areas of extrinsic thalamic nuclei (primary cortical areas) whereas in the neocortex of higher forms the projection areas (secondary cortical areas) of the intrinsic thalamic nuclei become dominant. The intrinsic thalamic nuclei are separable into two groups. Those projecting upon the neocortex become dominant in primates, whereas the intrinsic nuclei projecting upon the rhinencephalic structures are on the whole best developed in macrosmatic mammals. The ventral thalamus consists of one subdivision (ventral lateral geniculate body) entirely independent of the endbrain and of a second subdivision (reticular complex) which projects upon a large number of cortical fields. The sparse and generalized - though spatially well organized - projection of the reticular complex provides a system apparently independent of the dorsal thalamic projections and capable presumably, of evoking generalized cortical activity. PMID- 18421831 TI - Diffuse projection systems: the integrative action of the thalamic reticular system. PMID- 18421832 TI - Potential phenomena in thalamus and cortex. AB - 1. The cortical response to stimulation of an afferent path consists of two parts, a specific response indicating projection of the afferent and a response of the alpha mechanism. The former of these under the influence of strychnine can be identified with the cortical spikes of paroxysmal activity. 2. The response of the geniculate and other simply arranged nuclei are monophasic spikes. The diphasic spikes of the cortex and elsewhere are inferred to be assignable to two different groups of cells related by synaptic passage. 3. The unit cell potential may be interpreted as the resultant of unequal depolarization during activity of the two ends of the cell dendritic and axonal. If this is so, polarization by external currents should modify or even reverse the potentials and this proves to be the case. The phenomena are comparable to the effects of polarization on nerve axons. 4. Three types of potential fields can be visualized of which different cell structures of the brain may be considered as variants. Plots of the potentials of these fields indicate what relation of potential form to structure may be expected to obtain. 5. The character of the relation between thalamus and cortex appears to be different for the specific response apparatus which is not spontaneously active expect in paroxysmal states and the regulatory apparatus which is normally spontaneously active but suppressed in paroxysm. It is not necessary to infer reverberating closed circuits to account for the activity in either system. PMID- 18421833 TI - Electrical activity of subcortical areas in epilepsy. AB - The electrical activity of the cortex and subcortex were studied in 22 patients with epilepsy by means of scalp electrodes and a multi-electrode needle which was placed in the depths of the brain with the aid of a stereotaxic instrument. The instrument was designed for man and coordinate measurements were obtained by studies on brains fixed in situ. In the present operative series the position of the needle was checked by pneumoencephalography at the end of the recording. Analysis of records from the scalp and depths of the brain lead to the following conclusions: 1. The cortex and sub-cortex in epileptic patients display comparable normal and abnormal activity. 2. Like the cortex, however, various areas of the sub-cortex may show entirely independent abnormal activity. 3. During the induction of pentathol anesthesia 20 to 30 per second activity appeared first and remained most prominent in outer subcortical leads and showed least in leads from the central grey mass. 4. Isolated seizure discharges from the cortex are common, whereas they are rare in the subcortex. The fact that they do occur is important, because they may account for therapeutic failure in some cases where a cortical focus is ablated. 5. No case was found in which seizure discharges in or around the medial thalamus could be interpreted as initiating 3 per second wave-and-spike discharges of the petit mal type. 6. Primary and secondary cortical discharges have negative sign when referred to a relatively indifferent area. An incoming volley from a distant area is registered as a positive disturbance if referred to an indifferent region. 7. Electrical sign has localizing value in electroencephalography: Negativity indicates a local disturbance; positivity indicates a distant disturbance. PMID- 18421834 TI - An analysis of the sensory responses of the acoustic cortex. PMID- 18421835 TI - Brain stem reticular formation and activation of the EEG. AB - 1. Stimulation of the reticular formation of the brain stem evokes changes in the EEG, consisting of abolition of synchronized discharge and introduction of low voltage fast activity in its place, which are not mediated by any of the known ascending or descending paths that traverse the brain stem. The alteration is a generalized one but is most pronounced in the ipsilateral hemisphere and, sometimes, in its anterior part. 2. This response can elicited by stimulating the medical bulbar reticular formation, pontile and midbrain tegmentum, and dorsal hypothalamus and subthalamus. The bulbar effect is due to ascending impulses relayed through these more cephalic structures. The excitable substrate possesses a low threshold and responds best to high frequencies of stimulation. 3. Some background synchrony of electrocortical activity is requisite for manifestation of the response. In the "encephale isole", reticular stimulation has no additional effect upon the fully activated EEG. With synchrony, in spontaneous drowsiness or light chloralosane anesthesia, the effect of reticular stimulation is strikingly like Berger's alpha wave blockade, or any arousal reaction. In full chloralosane anesthesia, high voltage slow waves are blocked but no increase in lower amplitude, fast activity occurs. With barbiturate anesthesia, the reticular response is difficult to elicit or is abolished. 4. In the chloralosane preparation, the secondary cortical response evoked by a sensory volley is generally unaffected by reticular stimulation. Consequent sensory after-discharge is abolished, however, as is pyramidal tract discharge and jerky movements referable to it. Outside the sensory receiving area, secondary responses themselves may be reduced or prevented. 5. The convulsive spikes produced by local strychnine and those of a fit following supramaximal cortical excitation, are not decreased by stimulating the reticular formation. 6. The cortical recruiting response induced by low frequency stimulation of the diffuse thalamic projection system is reduced or abolished by reticular stimulation. 7. There is some indication that the cortical effect of reticular stimulation may be mediated by this diffuse thalamic projection system, for synchronized activity within it is similarly prevented by reticular excitation, and direct high frequency stimulation of this system, within the thalamus, reproduces the reticular response. It is possible, however, that other mechanisms may be involved in its mediation. 8. The reticular response and the arousal reaction to natural stimuli have been compared in the "encephale isole", in which EEG synchrony was present during spontaneous relaxation or was produced by recruiting mechanisms, and the two appear identical. 9. The possibility that the cortical arousal reaction to natural stimuli is mediated by collaterals of afferent pathways to the brain stem reticular formation, and thence through the ascending reticular activating system, rather than by intra-cortical spread following the arrival of afferent impulses at the sensory receiving areas of the cortex, is under investigation. 10. The possibility is considered that a background of maintained activity within this ascending brain stem activating system may account for wakefulness, while reduction of its activity either naturally, by barbiturates, or by experimental injury and disease, may respectively precipitate normal sleep, contribute to anesthesia or produce pathological somnolence. PMID- 18421837 TI - Sensory stimulation during sleep. Observations on the EEG responses to auditory stimulation during sleep in patients with brain pathology (Preliminary report). AB - 1. Potentials evoked by auditory stimulation during sleep were studied on a group of patients with unilateral brain damage resulting from external trauma cerebral vascular accident and tumor. 2. In a normal control group the response patterns from homologous areas in the two hemispheres were almost identical (except for the temporal leads). 3. In contrast with this, responses from damaged areas show distinct distortion of the pattern. 4. Records of typical cases are presented. PMID- 18421836 TI - Effect upon the EEG of acute injury to the brain stem activating system. AB - The effect upon the EEG in the unanesthetized "ecephale isole" of acute brain stem lesions in a position to involve the ascending reticular activating system has been studied. Elimination of the bulbar segment was without marked effect. Some synchronization followed elimination of the pons, but the most pronounced and prolonged changes occurred as a result of mesencephalic transection, or of discrete injury to the midbrain tegmentum or basal diencephalon, following which the EEG activation pattern of low voltage fast activity was reduced or abolished and the cortical record became dominated by recurring bursts or spindles of high voltage slow waves like those of normal sleep or barbiturate anesthesia. Bursts could be recorded from the intralaminar and other nuclei of the thalamus and these thalamic bursts were abolished by acute decortication. Conversely, cortical bursts were abolished by acute thalamic lesions. Possible interrelations of these regions in this activity is discussed. These results offer an explanation for the clinical observation of somnolence following basal injury to the brain, and suggest that a maintained influence of the ascending brain stem activating system underlies wakefulness, while absence of this influence precipitates sleep. PMID- 18421838 TI - Electroencephalographic study of experimental cerebro-vascular occlusion. AB - It was possible to follow the changes in the electroencephalogram during a protracted period in a group of 29 dogs in whom the middle cerebral artery had been clipped to its origin. A slight variation in the location of the clip was unavoidable but useful in that the size of the resulting infarcts differed. Massive lesions included the cortex in the fronto-temporo-parietal region, the striatum, the internal capsule and the anterior portions of the thalamus and, rarely, the hypothalamus. The majority of lesions were confined to the internal capsule and to the caudate nucleus. The clinical syndrome consisted of a contralateral spastic hemiparesis with the loss of the placing reaction and of position sense. Forced circling movements occurred towards the side fo the lesion, persisting even after blindfolding the animal. The changes in the electroencephalogram varied with the size of the lesion. In the scalp to ear records, slow frequencies were most conspicuous over the involved hemisphere in all types of recordings and were most prominent in recording to the contralateral ear. In the scalp to scalp records, flattening was evident over the frontal and temporal regions and slow frequencies were conspicuous posteriorly. Flattening was found over the entire hemisphere only in the most massive lesions and then in both types of recordings. The scalp to scalp records tended to become more symmetrical after two to three weeks, the flat tracings being replaced at times by high voltage slow frequencies. The abnormalities in the scalp to ear records tended to persist for the duration of the experiment. Flat records were obtained in the electrocorticogram overlying the area of infarction, and high voltage slow waves were found in the peripheral zone. Using the multipolar needle electrodes, an unusually low voltage to flat records was obtained from the depths of the lesion. A fairly constant change was the loss of the high voltage 10-12 per sec. spindles from the fronto-central scalp electrodes and from electrodes placed on the sigmoid gyri. The sleep records were far superior to the waking records as a diagnostic aid, the latter being of value only during the first post-operative week. PMID- 18421839 TI - A method for the rapid determination of the degree of forced breathing. PMID- 18421840 TI - EEG observations in a case with thalamic syndrome. AB - A case of thalamic syndrome of the Dejerine-Roussy type is presented. EEG studies reveal that although bilaterally symmetrical rhythmic activity may be present when the patient is awake, during drowsiness and deeper sleep a marked asymmetry often appears with early disappearance of alpha activity and reduction or abolition of spindles on the side involved. Thalamic integration in the sleep mechanism is briefly discussed. PMID- 18421841 TI - Brief summary of analyses of the 2007 survey on the capacity of National Influenza Centres. PMID- 18421842 TI - Methods molecular biology. Preface. PMID- 18421843 TI - [Diversity in mechanism of angiotensin II production and application of the suppressive agents]. PMID- 18421844 TI - [Clinical trials of ACE inhibitors and ARB for treatment of patients with hypertension, heart failure, and diabetic nephropathy in Japan--special reference to the dosage schedule and adverse effects]. PMID- 18421845 TI - [Clinical trials of renin-angiotensin antagonists in Japan]. PMID- 18421846 TI - [Clinical pharmacology of renin-angiotensin system]. PMID- 18421847 TI - [AMPA receptor]. PMID- 18421848 TI - [Therapeutic effect of clioquinol for Alzheimer disease]. PMID- 18421849 TI - [Therapeutic effect of S1P receptor modifying drugs for autoimmune diseases]. PMID- 18421850 TI - [Study on A-922G genetic polymorphism of endothelial nitric synthase associated ischemic stroke]. AB - OBJECTIVE: To explore the relationship between A-922G genetic polymorphism in the 5' flanking region of the endothelial nitric oxide synthase (eNOS) gene and ischemic stroke. METHODS: A 1:1 matched case-control study was conducted. 309 patients suffered from ischemic stroke were selected from two general hospitals in Shenzhen. The controls were selected by the same gender and ethnic group, and each pair's ages were permitted of differ within 5 years. eNOS A-922G genotypes were determined by using the real-time PCR technology for SNP genotyping with Taqman MGB probes. The cases and controls were interviewed by using the same questionnaire. Results The genotype frequencies (AA, AG and GG) of eNOS A-922G were 78.96%, 17.80, and 3.24% in patients, and 85.11%, 13.59, and 1.29% in controls. The allele frequencies of eNOS--922G in case group (12.14%) were higher than those in the controls (8.09%) (P = 0.018). OR value of ischemic stroke for subjects with at least one G allele was 1.523 (OR 95% CI 1.005-2.309). Results of conditional logistic regression demonstrated that eNOS A-922G genetic polymorphism also had significant effects on ischemic stroke (OR = 2.156, 95% CI 1.081-4.299) after adjusting smoking, alcohol drinking, waist-to-hipratio, body mass index and history of hypertension, et al. CONCLUSION: It was suggested that A-922G gene polymorphism of eNOS gene could associate with the higher risk of ischemic stroke susceptibility. PMID- 18421851 TI - [Application of double created restriction site PCR-RFLP to identify MGMT gene polymorphisms]. AB - OBJECTIVE: To develop a proper assay for identifying single nucleotide polymorphisms( SNPs) of the MGMT gene. METHODS: PCR primers were designed by create restriction site (CRS) method, then polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was adopted to identify four SNPs in MGMT gene. RESULTS: By PCR, one primer pair yielded target products containing MGMT84 SNP site, and the other primer pair yielded target products containing MGMT143, 160, 178 SNP sites. Four restriction enzymes were adopted to identify the four SNPs, respectively. The effects of PCR and RFLP were good. CONCLUSION: The methods for four SNPs of MGMT determinated by CRS-PCR-RFLP theory could be facility, economy, and rapidness. PMID- 18421852 TI - [Effect of dizocilpine maleate and taurine on the circuitry of glutamate and glutamine in rat striatum exposed by manganese]. AB - OBJECTIVE: To observe the effect of MK-801 and taurine on the activities of GS and PAG and the glutamatic neuron content in the rat striatum exposed by manganese. METHODS: 40 Wistar rats were random divided into four groups. The first group was the control group which was subcutaneously injected at the content of 0.9% NaCl. The second group was MnCl2 group which was subcutaneously injected at the content of 0.9% NaCl. The third and fourth groups were pretreatment groups which were subcutaneously injected of 0.3 micromol/kg MK-801 and 1 mmol/kg taurine. After 2h, the first group was peritoneally injected at the content of 0.9% NaCl, the 2nd-4th group were peritoneally injected at the dose of 200 micromol/kg MnCl2. All administration was given at the dose of 5 ml/kg for 25d, the pretreatment groups were given for every other day. At the 24h after the administration of MnCl2, the 4 rats brain tissue in every groups were removed after the rats were perfused from the left ventricles to with 4% ploymerisatum. The glutamatic neuron content were determined by immunohistochemical method (SABC). The activities of GS and PAG in the residual rat striatum were determined. RESULTS: In comparison with control group, the percentage of positive area and integral optical densities of glutamate immunocreative cell increased significantly in MnCl2 group (P < 0.01), the activity of GS decreased significantly in MnCl2 group, the activity of PAG increased significantly in MnCl2 group. In comparison with MnCl2 group, the percentage of positive area and integral optical density of glutamate immunocreative cell decreased significantly in MK-801 and taurine pretreatment group, the activity of GS increased significantly in MK-801 and taurine pretreatment group , the activity of PAG decreased significantly in MK-801 pretreatment group. CONCLUSION: MK-801 and taurine have a certain protective effect on "the circuitry of glutamate and glutamine" disrupted with Mn. PMID- 18421854 TI - [Comparative study of the effect on oxidative damage in rats inhaled by nano sized and micro-sized silicon dioxide]. AB - OBJECTIVE: To compare the changes of oxidative damage in rats inhaled by nano sized and micro-sized silicon dioxide. METHODS: 36 male rats were randomly divided into two control groups and four experimental groups which was inhaled by nano-sized and micro-sized silicon dioxide respectively at the concentration of 300 mg/m3 for 2 hours and the activities of SOD, CAT and GSH-Px and the contents of H2O2, GSH and MDA of the liver, kidney, brain were determinated 24h and 48h after inhalation. RESULTS: The contents of H2O2 in all organs in nano-sized groups increased significantly while increased in micro-size groups only in liver and kidney at 24h and in brain at 48h. On the contrary, the activities of CAT in nano-sized group were lower than those in micro-sized group. Superoxide anion contents increased only in the brain of nano-sized group. The activities of SOD decreased significantly in nano-sized groups in kidney at 24h and brain but not in micro-sized groups. The content of GSH decreased only in liver at 24h. The activities of GSH-PX decreased significantly in nano-sized compared with control group and were lower significantly in brain than those in micro-sized group. The contents of MDA increased in all nano-sized groups but only in liver and brain in micro-sized group. The total anti-oxygen activities decreased in all nano-sized groups, but only in kidney at 24h and brain in micro-sized group, especially more significantly in brain at 48h in nano-sized than micro-sized group. While the activity in kidney in nano-sized group increased at 48h comparing to at 24h. CONCLUSION: Nano-sized silicon dioxide could induce oxidative damage more easily than micro-sized silicon dioxide. Through comparing different interval, it was found that the degree of oxidative damage at 48h after inhalation inferior to that at 24h after inhalation, which could be associated with the repair of the body against the oxidative damage. PMID- 18421853 TI - [Study of C-ZnO composite nanoparticles on viability inhibition and DNA damage of mouse embryo fibroblasts]. AB - OBJECTIVE: To study the cytotoxicity and DNA damage induced by C-ZnO composite nanoparticles in Mouse embryo fibroblasts(MEF) cells. METHODS: MEF cells were treated with the suspension of carbon nanoparticles, zinc oxide nanoparticles and composite nanoparticles at the concentrations of 5, 10, 20, 50, 100 microg/ml. Cellular morphology and MTT assay were carried out to evaluate the viability of cells treated with particles for 24 h, 48 h and 72h respectively. Single cell gel electrophoresis (SCGE, comet assay) was performed to assess DNA damage induced by nanoparticles at the doses of 5 microg x ml(-1) and 10 microg x ml(-1). RESULTS: Three types of nanoparticles could induce morphologic change and viability inhibition with significant logarithmic time-effect and dose-effect relationship. Median lethal concentrations (IC50) of carbon nanoparticles, zinc oxide nanoparticles and composite nanoparticles were 21.85, 21.94 and 461.10 mcirog/ml respectively. The inhibitory effect on cellular viability of Zinc oxide nanoparticles increased significantly with the dose enhances. The most DNA damage was induced by composite nanoparticles at lower concentrations. CONCLUSION: Nanoparticles with different chemical composition induced distinct cytotoxicity and DNA damage in MEF cells. The metal content of composite nanoparticles played an important role in the toxic effect. PMID- 18421855 TI - [Study of the arsenate reducted by common reducing agents]. AB - OBJECTIVE: To study the role of glutathione (GSH), cysteine (Cys), vitamin C (VC) and ferrous ion (Fe2+) in the reduction of arsenate [As (V)] and the characteristic of reduction. METHODS: The reaction conditions [(37 degrees C, pH7.0 phosphate buffer) and the concentrations of As (V) (3 micromol/L), GSH (60 mmol/L), Cys (60 mmol/L), VC (0-30 mmol/L) and Fe2+ (0-40 mmol/L)] were simulated in vitro. Arsenite[As(III)] and [As(V)] were determined by the high performance liquid chromatography-hydride generation-atomic fluorescence spectrometry (HPLC HG-AFS). RESULTS: The reduction, of As(V) and As(III) formation, increased with the concentration, of GSH (0-60 mmol/L), and reached a maximum values at the concentration of 60 mmol/L GSH, then decreased with the concentration of (60-80 mmol/L) GSH. -SH in sulfhydryl compound, could play an important role in the reduction of As(V), and at the concentration of 60 mmol/L -SH (both GSH and Cys) could reduce 60%, of As(V) to As(III) for 30 min. VC or Fe2+ could not reduce As(V) alone, but when the concentration of VC or Fe2+ reached to 10 mmol/L or 20 mmol/L, they could have a significantly synergistic effect with 60 mmol/L -SH on the reduction of As(V) (P < 0.01). CONCLUSION: SH could reduce As(V) alone without any reductase. VC and Fe2+ could have a synergistic effect with -SH on the reduction of As(V). PMID- 18421856 TI - [Change of chart genes expression in small intestines of mouse induced by electromagnetic pulse irradiation]. AB - OBJECTIVE: To explore the bio-effects of electromagnetic pulse(EMP) on mouse small intestines induced by means of gene chip. METHODS: Twelve BALB/c mice were randomly assigned to the normal control group and the EMP group with 6 in each group. The EMP group was irradiated with 200 kV/m, 200 pulses EMP. 18 hours after the irradiation, the mice were sacrificed and their jejunum of small intestines were eviscerated. The fluorescent cDNA probes labeled with Cy3 and Cy5 were prepared from RNA extracted from the intestines of the two groups. Probes of the two groups were then hybridized against cDNA gene chip, the fluorescent signals were scanned with a scanner and the results were analyzed by computer. RESULTS: Compared with the control, 56 genes in gene expression profile were altered. The expression levels of 37 genes were up-regulated distinctly while 19 genes were down-regulated significantly. Among the 56 genes, 19 were reported with known or inferred functions, 12 up-regulated genes were catenin alpha 1 (alpha-catenin), ly-6 alloantigen(Ly-6E), fructose-6-phosphate transaminase (GF6P), ribosomal protein S17 (rpS17), small proline-rich protein 2A (Sprr2a), glandular kallikrein27 (GK27), lipoxygenase-3, aldo-keto reductase (Akr1c12), GSG1, amylase 2 (Amy2),elastase 2, p6-5 gene and 7 down-regulated genes were junctional adhesion molecule (Jam), protein arginine methyltransferase (Carm1),NNP-1, 2-5 A synthetase L2,Mlark gene, ATP synthase alpha subunit, uncoupling protein-2 (Ucp2) gene; the other 37 were reported with unknown functions. CONCLUSION: EMP irradiation could induce specific expressions of some genes in mouse small intestines and most of these genes were up-regulated ones. PMID- 18421857 TI - [Effects of microwave irradiation on NMDA receptor subunits mRNA expressions in rat hippocampus]. AB - OBJECTIVE: To study the role of NMDA receptor in learning and memory deficit caused by microwave irradiation. METHODS: At the time of Oh, 3h, 12h, 24h, 3d after microwave irradiation of 65mW/cm2 exposure for 20min (SAR 12.0 W/kg), rats were decapitated under deep anesthesia, the hippocampus was quickly removed, then expressions of NR1, NR(2A), NR(2B), NR(2C), NR(2D) subunits of NMDA receptor were measured by RT-PCR. RESULTS: Compared with control, the mRNA expressions of NR1 decreased in 21.1% (P < 0.05),and in 14.2% (P < 0.05), and in 30.3% (P < 0.01) respectively at timepoints of 3h, 24h, 3d after microwave exposure. At timepoints of Oh, 3h, 12h after microwave exposure, the mRNA expressions of NR(2A) decreased in 37.0% (P < 0.01), and in 35.9% (P < 0.05), and in 35.3% (P < 0.05) respectively. The mRNA expressions of NR(2B) did not statistically change after microwave exposure. The mRNA expressions of NR(2C) decreased in 24.3% (P < 0.05), and in 27.1% (P < 0.05) respectively at timepoints of Oh, 24h after microwave exposure. At timepoints of Oh, 12h, 24 h, 3 d after microwave exposure, the mRNA expressions of NR(2D) increased in 67.9% (P < 0.05), and in 45.7% (P < 0.05), and in 49.7% (P < 0.01), and in 75.4% (P < 0.01) respectively. CONCLUSION: The results indicated that the composition of subunits comprising NMDA receptor could be altered and autoregulation of NMDA receptor in the rat hippocampus exposed by microwave could decrease. These changes could be correlated with impaired LTP induced by microwave irradiation. PMID- 18421858 TI - [Effect of freeze-dried sea cucumber powder of eastern sea on tumor and immune index of S180-bearing mouse]. AB - OBJECTIVE: To study the effect of freeze-dried sea cucumber powder from eastern sea on tumor and immune of S180-bearing mouse. METHODS: S180 tumor cells were subcutaneouly inoculated to fifty health kunming mice and were randomly divided into five groups (model control group, positive control group, and low, medium, high dose groups). After the groups were orally treated with solution of freeze dried sea cucumber powder for ten days,the weights of tumors and the inhibition rates of tumor were counted, and the pathological changes of tumors were observed and the expressions of caspase-3 proteins were detected in the tumor tissues, and the thymus index and spleen index were observed , the contents of interleukin 2(IL-2) in the serum were detected. RESULTS: (1) In comparison with the control groups,the weights of tumor were declined and the inhibition rates of tumor were remarkably decreased in the treated groups. (2) The pathological changes of model control group were very obvious. The expressions of caspase-3 proteins in the treated group were more higher than those of model control groups, while were more lower than those of positive control groups. (3) The thymus index, spleen index in the treated group indicated an increased tendency. (4) The contents of IL-2 in the treated groups were more higher than those of other groups. CONCLUSION: Freeze-dried sea cucumber powder could be effective antitumor and could protect and recovery the immune system of S180-bearing mice. PMID- 18421859 TI - [Physical activity status of working time and its change over a ten-year period in Beijing and Guangzhou populations]. AB - OBJECTIVE: To describe the current status of physical activity of working time and its changes in the past ten years in certain Chinese populations. METHODS: 3304 participants were randomly selected in clusters (villages, residential households, or working organizations) from 4 approximately equally sized sub samples, an urban and a rural district in Beijing and an urban and a rural district in Guangzhou. Physical activities of working time and its changes in the past ten years were collected by a standard questionnaire. The energy expenditures per kilogram body weight (unit: kJ/kg) in one normal working day for each individual were calculated. RESULTS: The physical activity levels were relatively low and the rates of very mild and mild physical activities were 59.4% in these four Chinese populations. Intensities of physical activities at working time were greater in men than those in women, and in southern than those in northern, and in rural than those in urban in China. As a whole, 48.8%, 38.0% and 13.2% of our study participants reported that physical activities of their working time could decrease, could unchange, and could increase respectively over the ten years period before the survey. The rates of decreases were greater in rural than those of urban, and greater in men than those of women, increased with age, and decreased with educational levels. CONCLUSION: Low physical activity at working time and its declining trends in these Chinese populations should require public health investments, including strategic planning and public education. PMID- 18421860 TI - [Study on laboratory analysis and its variety for calcium in prepackaged foods]. AB - OBJECTIVE: To observe the variety of calcium analysis of prepackaged foods from different labs of China. METHODS: Fourteen prepackaged foods from seven kinds were selected as food samples including cereal products, bean products, meat products, dairy products, beverages, nuts, snacks, according to national prepackaged foods consumption survey results in 2000. The calcium of samples were parallelly analyzed for three times by national standard methods in seven labs from all the country. RESULTS: The 78.6% of results were up to grade by Grubbs, Cochran and Dixon screening methods. The accuracy of protein analysis was expressed by variety co efficiency (C.V%). The C.V% of inner labs and inter labs were from 2.21% to 8.13% and from 5.51% to 31.12%, the total and average C.V% level were 5.93%-31.24% and 22.10%. Besides fish tin (with low calcium content), the relative expanded uncertainty of protein detection was from 10.36% to 57.10% (average 39.78%). CONCLUSION: Calcium analysis level in labs from all the country was fit for national standards, the most of variety and uncertainty of analysis was less than 60%. PMID- 18421861 TI - [Study on SOCS-3, PPARgamma and ACO mRNA expression of adipose tissues in obese rats with leptin resistance]. AB - OBJECTIVE: To study the difference of socs-3, PPARgamma and ACO mRNA expression in obese rats with leptin resistance induced by high-fat diet. METHODS: 6 Wistar male rats were fed with basic diet as control, and 24 rats were fed with high-fat diet as high-fat diet group. At the end of the 8th week, 8 obese rats were selected by body weight from high-fat diet group as obese group. Serum leptin, socs-3, PPARgamma and ACO mRNA were detected. RESULTS: In compare with control group, body weight, serum leptin, socs-3 and PPARgamma mRNA expressions of epididymal adipose tissues were significantly increased in obese group, while ACO mRNA were markedly decreased. CONCLUSION: Obese rats induced by high-fat diet showed the suppression of leptin signaling transduction pathway, decreases in fatty acid oxidation and increases in fat synthesis. PMID- 18421862 TI - [Molecular mechanism by which green tea and tea extract inhibits left ventricle hypertrophy induced by renovascular hypertension in rats]. AB - OBJECTIVE: To investigate the preventive effects and the molecular mechanism of tea and tea extracts (polyphenols and EGCG) on left ventricle hypertrophy (LVH) induced by Renovascular hypertension in rats. METHODS: One hundred and sixty male Wistar rats were randomly divided into five groups: negative control group (SHAM), operation group (2K1C), green tea group (2K1C + GT), tea polyphenol group (2K1C + TP), EGCG group (2K1C + EGCG). After surgery operation, rats in different groups received tap water, tap water, 2% green tea, 0.1% tea polyphenol and 0.05% EGCG respectively, as the sole drinking source for 8 weeks until the end of the experiment. RESULTS: The results showed that the blood pressure, the ratio of left ventricle weight to body weight (LVW/BW) and the left ventricular wall thicknesses (LVWT) in 2K1C group significantly increased (P < 0.01), when compared to those in sham operation group. However, when compared to these parameters in operation group, the ratio of LVW/BW and HW/BW in green tea group, tea polyphenols group and EGCG group significantly decreased (P < 0.05), while the activities of GSH-Px and SOD increased, and the levels of ROS in heart were also significantly decreased (P < 0.05). Ras, P-ERK protein expressions in tea treatment groups were also decreased. CONCLUSION: Green tea, tea polyphenols and EGCG can attenuate the development of left ventricular hypertrophy induced by renal hypertension in rats. The possible mechanisms may be due to its antioxidant properties and the modulation of Ras-to-MAPKs mediated signal transduction. PMID- 18421863 TI - [Technic study on multilocus sequence typing system of Salmonella enteritidis]. AB - OBJECTIVE: To characterize different Salmonella enterica Enteritidis isolates through multilocus sequence typing (MLST) system, a molecular epidemiologic analysis method was established. METHODS: Internal fragments of six housekeeping genes, thrA, purE, sucA, aroC, hemD and dnaN and a unique gene of S. Enteritidis were amplified and sequenced to develop the MLST method. Eighteen S. Enteritidis food isolates with unique serotype were studied and the data were analyzed by the Sequener 4.0 software. The genetic relationship of the isolates was estimated, and the DNA sequence diversity was also studied between S. Enteritidis and S. Typhimurium LT2. RESULTS: A poor DNA sequence diversity among S. Enteritidis 50041 and 18 S. Enteritidis isolates in all six housekeeping genes were found. The number of variable sites per gene ranged from 1 to 6 nucleotides between S. Typhimurium LT2 and S. Enteritidis. Furthermore, S. Enteritidis 50041 and 18 S. Enteritidis isolates contained a mutation at C182T in SdfI in comparison with to the sequence in the GenBank. Our study showed that the MLST had poor discrimination power among S. Enteritidis isolates. CONCLUSION: The MLST techniques should only be used to differentiate Salmonella isolates with different serotypes, it could not study the molecular epidemiological relationship of the isolates with the same serotype. PMID- 18421864 TI - [Immunological analysis and mass spectrometry identification of the major allergen from Cladosporium cladosporioides]. AB - OBJECTIVE: To analyze and identify the major allergen from Cladosporium cladosporioides. METHODS: The Cladosporium cladosporioides were collected by means of glass dish exposed by air. After they were cultured, the protein of the Cladosporium cladosporioides was extracted with carbonate buffer solution. The major allergen in the Cladosporium cladosporioides was characterized by Western blotting after SDS-PAGE. After in-gel digestion, the peptide mass values of the major allergen which were obtained by matrix-assisted laxer desorption ionization time-of-flight mass spectrometry, (MALDI-TOF-MS) were used as inputs to search corresponding proteins with the Mascot (http://www. matrixscience. corn). RESULTS: There were fourteen protein bands visible on the SDS-PAGE gel in the crude extracts. The most abundant ten bands were 72 kD, 61 kD, 42 kD, 36 kD, 35 kD, 34 kD, 24 kD, 22 kD, 18 kD and 14 kD proteins, respectively. The protein whose molecular mass was 18 kD was identified by Western-blotting as major allergen. The major allergen was similar to the 18 kD ribosomal protein of Psychrobacter arcticum based on peptide mass fingerprint analysis (MALDI-TOF-MS). CONCLUSION: The major allergen from Cladosporium cladosporioides, a ribosomal protein about 18 kD, was analyzed and identified. PMID- 18421865 TI - [Evaluation of two-year follow-up of group intervention on smoking cessation based on social cognitive theory]. AB - OBJECTIVE: To assess the effect of a group intervention on smoking cessation based on social cognitive theory. METHODS: A total of 217 smokers were recruited through mobilization in the community. Six-month and 2-year follow-up surveys were engaged after intervention to evaluate the effectiveness of the training. RESULTS: Smoking quitting rate confirmed by biomarker level in the 2-year follow up survey could achieve 32.3% and the successful quitting rate could be 15.2%. The factors influencing smoking cessation at 2-year follow-up could be associated with diseases related to smoking (AOR = 4.09), the change in self-efficacy scores (AOR: 1.05-1.13) and home smoke-free policy (AOR= 5.07). CONCLUSION: The group intervention on smoking cessation based on social cognitive theory could be effective and can be used in community. PMID- 18421866 TI - [Structural equation model analysis of relationship between occupational stress and work ability]. AB - OBJECTIVE: To construct the structural equation models of occupational stress and work ability to analyze the factor influencing work ability. METHODS: A test of occupational stress and work ability was carried out with revised occupational stress inventory (OSI-R) and work ability index (WAI) for 2041 workers. And structural equation model was used to analyze their relationship. RESULTS: The main factors influencing the work ability included the role insufficiency, psychological strain, physical strain, vocational strain. Rational/cognitive, education, income, recreation and self-care had some influence on the work ability, and they correlated positively with the work ability. And age, work age, role ambiguity and physical environment correlated inversely with the work ability. CONCLUSION: The work ability of physical laborers can be improved with adjusting occupational stressor and improving the status of maladjustment, and the structural equation model can be used to study occupational medicine. PMID- 18421867 TI - [Influence of family income on dietary nutrients intake and dietary structure in China]. AB - OBJECTIVE: To analyze the influence of family income on food consumption. METHODS: The paper used the food information based on 24-hour dietary recall method for 3 consecutive days and weighting method and family income information and analyzed the data with SAS software. RESULTS: Protein intake, fat intake, percentage of energy provided by fat, percentage of energy and protein coming from animal foods and some micro-nutrients intake had increasing trends with the increase of family income for Chinese urban and rural residents. CONCLUSION: Family income was an important factor to affect dietary nutrients intake and dietary structure of Chinese residents. PMID- 18421868 TI - [Analysis of health selective survey result of children and pregnant/Lying-in women in China in 2006]. AB - OBJECTIVE: To gain a information of the current status of the health condition of children and pregnant/lying in women in China, and provide evidence to make the relative police of the state. METHODS: The samples of national representative value were drawn by means of random sampling in layered groups, and recommended standards of WHO were used as the evaluation standard for children' s growth and pregnant/lying-in women's health condition, of the health related information of sample population were got by questionnaire survey and health examination and laboratory test, and statistics and analysis were used with SAS software. RESULTS: As of time the rates of low body weight of mid and high levels and slow growths in children under the age of 5 year old were 5.9% and 9.9%. Vitamin A deficiency rate in children under the age of 5 year old were 9.1% and the brink deficiency rate of vitamin A were 41.8%. Knowledge coverage rate in parents of under the age of 3 year old were 65.8%. Occurrence rates of in news born were 4.59%, anemia in pregnant/lying-in women were 42% in which anemia in lying-in women and in pregnant women respectively were 49.7% and 37.7%. CONCLUSION: Comparing with the survey result in 2002. malnutrition rates in children under the age of 5 year old are getting on the fall, and occurrence rates of new-born are getting rising, and anemia rates in pregnant/lying-in women were high level. PMID- 18421869 TI - [Study on the formulas predicted for body fat of children and adolescents by using bioelectrical impedence analysis]. AB - OBJECTIVE: To modified the prediction formulas for bioelectrical impadance analysis on the fat free mass at different body mass index (BMI) levels of Chinese children and adolescents. METHODS: Data of body composition of 468 boys and 457 girls aged 7-18 years living in Beijing measured by under water weighing were used to develop the equations of FFM for bio-electrical impedance. RESULTS: The prediction formulas were:normal weight group: FFM(kg) = 0.406 weight + 2.918 sex + 0.315 H2/z + 0.843. over weight group: FFM(kg) = 0.358 weight + 1.571 sex + 0.358 H2/z + 0.603. Total population: FFM(kg) = 0.290 weight + 2.222 sex + 0.427 H2/z + 1.547. The regression equations with the corresponding explained variance (r2) and the standard error of estimate (SEE) in normal, over weight and total population group are 0.940, 2.36, 0.955, 2.46 and 0.942, 2.53 respectively. The significant of the regression were tested by the ANOVA (P < 0.001). CONCLUSION: The explained variance and the prediction error of the formuals were improved compared to the literature by using,the classification screening standard of age- and sex-specific BMIs for overweight and obesity of Chinese school children to divide groups. PMID- 18421870 TI - [Cross-sectional survey of children body fat and influence analysis in Beijing and Guangzhou]. AB - OBJECTIVE: Knowing the percent of the body fat in the Chinese young and doing some basic works of making the fat mass criteria of the Chinese children obesity and prevention of the obesity in the young children. METHODS: 2445 children (1257 males and 1188 females) from Beijing and Guangzhou were studied. All of them underwent anthropometrical measurements and bioelectrical impedance analysis (BIA). By applying Chinese children fat mass predict equation to estimate their FFM. The percent of fat mass was calculated by subtracting FFM from weight (Wt). RESULTS: The body fat percent increase with the age increase both in boys and girls. There is gender different between boys and girls. The boys' BF(%) is higher than in the girls'. The boys and girls' BF(%) in Beijing are higher than boys and girls' BF(%) in Guangzhou. At the same growth degree with the ages increase the BF(%) decrease; while at the same age, with growth increase, BF(%) decrease both in boys and girls. CONCLUSION: The BF(%) is different between the ages, area, and gender and growth degree. PMID- 18421871 TI - [Dietary supplement use in Beijing]. AB - OBJECTIVE: To assess prevalence of dietary supplement use among Beijing population. METHODS: By stratified cluster sampling method, 4561 subjects in 6 different regions in Beijing were investigated distributed. Questionnaire survey was used to collect the information on dietary supplements use and the demographic, health and lifestyle characteristics. RESULTS: The prevalence of dietary supplements use in Beijing were 11.9% (standardized rate: 10.1%), 9.0% in male persons(standardized rate: 7.7%), and 14.5% in female persons (standardized rate: 12.4%), respectively. Main types of dietary supplements used were compounds multivitamin and multiminerals. The distributions of dietary supplement use were different in population with different characteristics. CONCLUSION: The prevalence of dietary supplement use was more less of the population in Beijng than that of the population in developed country. Further investigation should be carried out to find the influence factors of dietary supplements use in China. PMID- 18421872 TI - [Detection of the expression level of S100C gene in lung cancer tissue by RT-PCR and construction and identification of prokaryotic expression vector]. AB - OBJECTIVE: To analyze the expression level of S100C gene in lung cancer tissue and to construct prokaryotic expression vector pET30a-S100C in order to select fusion antibody in further study. METHODS: mRNA expression level of S100C gene in lung cancer tissue and ambient tissue in 26 cases was detected by RT-PCR with p actin as references. The full-length cDNA of S100C was cloned by RT-PCR. The amplified DNA fragments were ligated into pET30a vector and then transformed into E. coli stain JM109. The positive clones were screened out and indentified. RESULTS: The expression level of S100C in lung cancer tissue was lower than that in ambient tissue. The difference between the two groups was statistically significant (P < 0.05). The prokaryotic expression victor of S100C was successfully constructed. The sequence homogenuity was up to 100%. CONCLUSION: The down-regulation expression of S100C gene probably takes important part in the carcinogenisis of lung cancer. PMID- 18421873 TI - [Primary research on detecting bacteria drug resistance by matrix-assisted laxer desorption ionization time-of-flight mass spectrometry]. AB - OBJECTIVE: To study the feasibility of bacteria drug resistance determinated by matrix-assisted laxer desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS). METHODS: 11 strains of 8 genera in drug resistant-bacteria separated from clinical specimens and the corresponding standard strains were detected by MALDI-TOF-MS, and the differences of the mass spectrums were analyzed. At the same time, 30 strains of Staphylococcus aureus separated from clinical specimens were detected by MALDI-TOF-MS and their antimicrobial susceptibility were tested. All bacteria were cultured under the same conditions. RESULTS: When drug-resistant bacteria determinated by MALDI-TOF-MS, it was efficient for Staphylococcus, especially the methicillin-resistant Staphylococcus aureus, but it was not efficient for other genera. CONCLUSION: MALDI-TOF-MS can be used to detect drug resistance of some bacteria. PMID- 18421874 TI - [Determination of bisphenol A in drinking water system components by liquid chromatography-tandem mass spectrometry]. AB - OBJECTIVE: To develope the determination methods for bisphenol A in drinking water system components. METHODS: 30 samples were quantitatively analyzed by liquid chromatography-tandem mass spectrometry (LC/MS/MS). RESULTS: Bisphenol A was linear in the range 0-100 ng/ml and the linear correlation coefficient reached 0.9994. The minimum detection limit was 5 microg/kg. The spiked recoveries were from 90% to 99%, and the RSD were between 1.8 -5.0%. Bisphenol A was measured in nine out of thirty samples. The detected ratio was 30% and the maximum was 1.82 mg/kg. CONCLUSION: The method was quick, sensitive and specific. PMID- 18421875 TI - [Preventive effects of 4 Se-enriched plants on rat stomach cancer induced by MNNG 2. Safety of different selenium resources for prevention of stomach cancer]. AB - OBJECTIVE: To guide the application of selenium (Se) in the prevention of human chronic diseases, the safety of different Se resources (selenite, Se-enriched garlic, Se-enriched broccoli, Se-enriched green kale and Se-enriched red kale) was investigated. METHODS: Ninety rats were fed the basal diet for a week, divided equally into the control, N-methyl- N'-nitro-N-nitrosoguanidine (MNNG) and seven Se treatment groups. Rats were daily given 15 mg/kg bw of MNNG for ten days except those in the control group, and rats in seven Se supplementation groups were daily given 75 microg/kg bw or 150 microg/kg bw sodium selenite solution, 150 or 300 microg/kg bw Se as of Se-enriched plant suspension by gavage for 17 weeks. Rats were sacrificed at the end of 18th week, the liver glutathione S-transferase (GST), serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities were detected. Rats were given with the basal diet and water freely during the experiment. The data was statistically analyzed by SPSS 12.0 software. RESULTS: All rats in Se 75 microg/kg bw of Se as sodium selenite group died within 6 weeks, the body weights of rats in 75 microg/kg bw of Se as sodium selenite group were significantly lower than those of rats in other groups, the serum AST and ALT activities of rats in 75 microg/kg bw of Se as sodium selenite group were significantly higher than those of rats in other groups, and only the Se supplementation of 75 microg/kg bw of Se as selenite led to rat liver pathological change. CONCLUSION: Selenite is at least 4 times as toxic as Se-enriched higher plants selected in this experiment. PMID- 18421876 TI - [Study on vitamin B1, vitamin B2 retention factors in vegetables]. AB - OBJECTIVE: To study the influence factors of vitamin B1, vitamin B2 losses in vegetables during cooking. METHODS: 12 types of fresh vegetables were selected and cooked by different cooking process, including roast, braise, fry, steam, scald and salt curing. vitamin B1 and vitaminB2 contents in raw and cooked samples were analysed respectively by GB/T 5009.84--2003 and GB/T 5009.85--2003 fluorometry. At the same time, weights of vegetables before and after cooking were recorded. Then CY and RF were calculated. RESULTS: The results showed that the retention rate in leguminous vegetables was 66%-75% for vitamin B1 and 85% 190% for vitamin B. Scalding could cause bigger loss of vitamin B1 in leafy vegetables and root vegetables, RF was about 50. The values of vitamin B2 RF in leafy vegetables for roasting was bettering than for scalding. For vitamin B2, the retention rate in root and cucurbitaceous vegetables was about 77-79. CONCLUSION: Vitamin B1 and vitamin B2 retention factors of roasting and steaming are more than that of frying and scalding. Temperature, time and variety of vegetable are the important influence factors of vitamin retention factors. PMID- 18421877 TI - [Application of carboxylated polystyrene latex particles in the detection of aflatoxin B1]. AB - OBJECTIVE: To study on rapied detection technique of aflatoxin B1. METHODS: Carboxylated polystyrene latex particles(CPLP) were prepared by soap-free polymerization using styrene and alpha-methyl crylic acid. The CPLP were sensitized with aflatoxin B1 oxime-BSA for the detection of aflatoxin B1. RESULTS: The minimum detect limits of sensitized latex were 5 ng/ml, CONCLUSION: The sensitized latex could applied to detect qualitative and half-quantitative aflatoxin B1 in fieldwork. The technique maybe minim, simpleness, rapidness and specificity. PMID- 18421878 TI - [A convenient way for autoregressive moving average model order determination: autoregressive moving average (n, n-1) modeling strategy]. AB - OBJECTIVE: To introduce the autoregressive moving average (ARMA)(n, n-1) modeling strategy as to a convenient way for the determination of ARMA orders. METHODS: ARMA(n, n-1) strategy was generally explained. A simulated ARMA(3,2) time series was generated and analized following this strategy. RESULTS: Combined with AIC information criteria and residue plot, the order of ARMA model was determined and complied well with the simulated data. CONCLUSION: ARMA(n, n-1) strategy may be convenient for the medical workers. PMID- 18421879 TI - [Analysis and evaluation of S-phenylmercapturic acid as a biomarker for benzene exposure]. AB - Since urinary S-phenylmercapturic acid concentration could be well related to benzene exposure, it could be a sensitive and specific biomarker of low-level benzene exposure. SPMA may be determined by HPLC, LC/MS, GC/MS and ELISA methods. The abroad methods of determination of urinary SPMA were reviewed in detail in recently years and the validation and application of urinary SPMA as a biomarker for occupation benzene exposure were evaluated. PMID- 18421880 TI - [Research progress on influencing factors and improvement measures of elderly anemia]. AB - Anemia in the elders is an important public health crisis which more attention should be paid to in the aging society. Anemia decreased physical performances, impaired cognitive functions, increased rates of hospitalization and mortality and affected quality of life. Causes of anemia in this population were complex as well as multiple. Chronic diseases maybe the most common reason of anemia in elders, but its mechanism was too complex; anemia caused by nutrition deficiency was clear and easily prevented and well treated. Improving nutrition status could benefit the reduction of anemia prevalence, and thus it was significant that the life quality of elderly persons could be improved and a healthy aging society could be promoted. PMID- 18421881 TI - [Application of stable isotopic technique in the study of tissue protein metabolism]. AB - Study of total protein metabolism can not explain the protein turnover of individual organ or tissue, so it may be necessary to study the tissue protein metabolism. In this paper, reviewed the principle, main methods and development of stable isotopic technique in the study of tissue protein metabolism were reviewed, and with the introduction of advantages and limitations of each method were introduced. PMID- 18421882 TI - [Protective role of lutein on light-damage of retina]. AB - Lutein may be a member of the xanthophy II of carotenoids. It could be a non provitamin A xanthophy II, which was different from other carotenoids. Lutein and zeaxanthin composed the macular pigments, which were found in the retina. The protective mechanism of lutein in the retina could possibly be in two important ways: the first function could relate to filtering blue light, and the second could consider to be the antioxidative activity that could quench and scavenge reactive oxygen species(ROS) induced by light. The structure and source of lutein, and association between lutein and retina macular pigment, and the role and mechanism of lutein in protecting the retina from light damage were reviewed. PMID- 18421883 TI - [Advances in the study of genetic engineering vaccine delivered by probiotics]. AB - Probiotics are defined as viable microorganisms that exhibit a beneficial effect on the health of the host when they are ingested. They have many beneficial effects for health, and are commonly recognized as safe. With the development of the protein expression technique for probiotics, the study of probiotics to be used as protein, antigen and antibody delivery vehicles becomes a new highlight. This paper used Lactobacillus as the delegate of probiotics, summarized the effect principle, relevant technique and advances of genetic engineering vaccine delivered by probiotics. PMID- 18421884 TI - [Advancement of the study of the mechanisms of cells invades by HCV]. AB - Hepatitis C virus may be an important human pathogen, it may infect human associated with hepatocytes invaded by HCV. The construction features of HCV and its receptor candidates and the mechanisms of cells invaded by HCV were reviewed. PMID- 18421885 TI - [Present research on disease burden of diarrhea illness]. AB - At present, many countries have developed researches on disease burden of diarrhea illness. The foreign surveys are comprehensively and continuously estimated from the prevalence aspect. The domestic surveys lay put stress on some enteric pathogen and lack systematization, which lead to reduce data utility. In order to provide a reference for acute disease burden in China, present researches at home and abroad are reviewed. PMID- 18421886 TI - The bus to Copan. PMID- 18421887 TI - Exploring moderators of the relationship between physical activity behaviors and television viewing in elementary school children. AB - PURPOSE: Previous research suggests that there is limited evidence to support a negative association between physical activity (PA) behaviors and television (TV) viewing time in children. The purpose of this study was to extend the research involving PA-TV viewing relationships and to explore potential moderators, including gender, ethnicity, weekday/ weekend behaviors, structured/unstructured activities, and seasonal variability. DESIGN: A 9-month longitudinal design, across one school year, with assessments every 3 months. SETTING: Elementary schools in the Vancouver and Richmond districts of British Columbia, Canada. Subjects. Subjects (N = 344; 47% female) were 9- to 11-year-old children who participated in a school-based PA initiative from September 2003 to June 2004. INTERVENTION: Not applicable. MEASURES: Assessments of PA were measured using the Physical Activity Questionnaire for Children. TV viewing time and structured PA were measured using a self-report questionnaire. ANALYSIS: Basic descriptives, Pearson r bivariate correlations and moderated multiple regressions with mean centered variables. RESULTS: No significant interaction effects were found for any of the proposed moderators. Null bivariate correlations are supportive of findings in previous literature. CONCLUSIONS: Our results did not find support for PA-TV viewing relations, regardless of gender, ethnicity, structured PA, and seasonal variability. PA interventions aimed at modifying sedentary behaviors, such as TV viewing, may not be warranted. PMID- 18421888 TI - A systematic review of interventions to increase awareness, knowledge, and folic acid consumption before and during pregnancy. AB - OBJECTIVE: We conducted a systematic review of studies designed to increase awareness of knowledge about, and consumption of folic acid before and during pregnancy. DATA SOURCES: Studies were identified from Cochrane Library, Medline, and the references of primary studies and reviews. STUDY INCLUSION AND EXCLUSION CRITERIA: Studies included randomized controlled trials, quasi-experimental interrupted time series studies, follow-up studies, case-control studies, and before-and-after studies, all of which were conducted between 1992 and 2005 on women ages 15 to 49 years and/or health professionals, evaluating awareness and/or knowledge and/or consumption of folic acid both before and after intervention. Studies were excluded if data were not presented both before and after intervention or were other outcomes than those mentioned here. DATA EXTRACTION: Data were extracted in relation to characteristics of studies, participants, interventions, and outcomes. DATA SYNTHESIS: Because of heterogeneity, we performed a narrative synthesis describing the direction and the size of effects. RESULTS: On average, women's awareness increased from 60% to 72%, knowledge from 21% to 45%, and consumption from 14% to 23%. CONCLUSIONS: Interventions had a positive effect on folic acid intakes before and during pregnancy, although the average usage reached less than 25%. PMID- 18421889 TI - Social support, network heterogeneity, and smoking behavior in women: the 10-town study. AB - PURPOSE: Examine the associations among social support, network heterogeneity, and smoking behavior in a large sample of Finnish female municipal employees. DESIGN: Cross-sectional observational study. SETTING: Workplaces in 10 towns in Finland. MEASURES: Smoking behavior and social support from one's partner, relatives, friends, supervisor, and coworkers was assessed by a questionnaire survey in 2000 to 2001. ANALYSIS: Logistic regression analysis. SUBJECTS: Total of 23,069 female employees (mean age = 45 years). Separate models for heavy smoking (> or = 20 cigarettes per day) were calculated for 4119 current smokers. The association between social support and ex-smoking was examined in 7352 ever smokers. RESULTS: After adjustment for various confounders, women who reported low heterogeneity of their support network or no support from their partners or relatives were approximately 1.2 times more likely to be smokers compared with their counterparts with high heterogeneity or high support. Low network heterogeneity was also significantly associated with an elevated prevalence of heavy smoking in current smokers (odds ratio [OR] = 1.52; 95% confidence intervals [CI] = 1.06, 2.19) and a lower likelihood of ex-smoking in ever-smokers (OR = 0.74; 95% CI = 0.63, 0.87). The associations were more pronounced among nonmanual vs. manual employees. CONCLUSION: Our findings suggest an association among network heterogeneity, sources of social support, and smoking in female employees. The preventive impact of these social resources on smoking behavior is stronger among nonmanual female employees. PMID- 18421890 TI - Faith-health collaboration in the United States: results from a nationally representative study. AB - PURPOSE: Estimate the prevalence of and identify characteristics associated with religious congregations' collaboration with health agencies. DESIGN: Cross sectional analyses of self-report data from the National Congregations Study, a random sample of religious congregations generated from the 1998 General Social Survey. SETTING: United States. SUBJECTS: Key informants from 1236 congregations. Each respondent described a single congregation. MEASURES: Respondents provided open-ended descriptions of congregational programs. Researchers coded program descriptions by content (e.g., domestic violence) and whether the program involved collaboration with a secular agency. Other congregational characteristics (e.g, denomination) were measured by validated measures and linked census tract data. RESULTS: Overall, 11.1% of congregations participated in faith-health collaboration (FHC). Logistic regression analyses found that FHC was more common among congregations with more members, with a small proportion of congregants under 35 years, and with a senior pastor with a graduate degree. Other effects were conditional; for instance, denominational differences varied depending on urban/suburban/rural location and the proportion of low-income members. CONCLUSION: This study provides the first national estimates of the prevalence of FHC. Such collaborative efforts may require different approaches in different areas. These results can help practitioners identify congregations that may be more willing to collaborate. PMID- 18421891 TI - Evaluation of a "nondieting" stress reduction program for overweight women: a randomized trial. AB - PURPOSE: Determine if a "nondieting" intervention focused on intensive training in eliciting the relaxation response enhances health outcomes compared with nondieting interventions without such training. DESIGN: Randomized trial with follow-up at 10 weeks, 4 months, and 12 months. SETTING: General community. SUBJECTS: Total of 225 overweight and obese women with at least one other cardiovascular risk factor. INTERVENTIONS: Three 10-week nondieting interventions: a group program (P1) focused on intensive training in techniques for eliciting the relaxation response (n = 60), a group program (P2) focused on healthy eating and physical activity (n = 61), and a self-guided, mail-delivered version of P2 (P3; n = 101). MEASURES: The Revised Symptom Checklist measured psychological distress, the Medical Symptoms Checklist measured the experience of medical symptoms, and the Health-Promoting Lifestyle Profile measured a range of lifestyle behaviors. Self-efficacy for low-fat eating intuitive eating, and body mass index were also assessed. ANALYSIS: An intention-to-treat analysis was used. RESULTS: At 12 months, P1 produced statistically greater improvements in stress management behaviors and medical symptom discomfort and was the only program to significantly improve self-efficacy for low-fat eating. In P1, the effect sizes for reductions in depression (0.75) and interpersonal sensitivity (0.85) were large. At 12 months, mean weight was unchanged. CONCLUSION: Inclusion of intensive relaxation response training in a nondieting program for overweight women enhanced stress management and medical symptoms outcomes but not weight outcomes. PMID- 18421892 TI - Evaluating a peer intervention strategy for the promotion of sexual health related knowledge and skills in 10- to 14-year-old girls. Findings from the "Entre amigas" project in Nicaragua. AB - PURPOSE: Report effects on knowledge of sexual health and gender from an intervention using peer methodology in Nicaragua. DESIGN: A prepost nonequivalent control group design. SETTING: Ciudad Sandino, Managua, Nicaragua. SUBJECTS: A total of 599 girls were surveyed, 60% nonintervened and 40% intervened. INTERVENTION: Peer methodology consisted of (1) meetings in which girls talked and worked with other girls, (2) mothers taking an active role in the peer groups, and/or (3) girls watching the soap opera "Sexto Sentido." MEASURES: Indices measuring changes in sexual knowledge and gender vision. RESULTS: Girls participating in the peer groups were twice as likely to have satisfactory sexual health-related self-esteem as those who did not participate. Eleven percent of the girls achieved satisfactory self-esteem as a result of the (peer groups x mothers) interaction and 15% due to the (peer groups x mothers x "Sexto Sentido") interaction. Girls participating in the peer groups were three times as likely to have satisfactory gender visions; if exposed to all three components, they were almost four times as likely to develop satisfactory gender visions. CONCLUSIONS: Peer methodology, participation of a female family member, and an educational soap opera seem beneficial in promoting sexual health-related knowledge and gender vision in young girls. PMID- 18421893 TI - Predictors of stage transitions in the precaution adoption process model. AB - PURPOSE: To explore psychosocial correlates and predictors of stage transitions in the precaution adoption process model (PAPM) for fruit intake. DESIGN: A cohort completed three electronic questionnaires, at baseline (time 0), 35 days later (time 1), and another 32 days later (time 2). SETTING: Secured Internet Web site. SUBJECTS: A cohort of 735 adults was formed from a random sample of an existing Internet panel. The mean age was 37.5 years, 51% were women, and 90% were of Dutch origin. Most respondents (48%) had a medium level of education. MEASURES: Precaution adoption process model stage, risk perception, perception of own fruit intake level, attitude, pros, cons, subjective norms, social support, modeling, self-efficacy, and fruit intake (assessed using a food frequency questionnaire). ANALYSIS: Cross-sectional differences in psychosocial variables and fruit intake across PAPM stages at baseline were analyzed using analysis of variance with Tukey multiple comparisons tests. Predictors of PAPM stage transitions between time 0 and time 1 and between time 1 and time 2 were analyzed using logistic regression analysis. RESULTS: Factors related to attitude and social influences may be important if one is to decide to act, whereas strong self-efficacy may also be required for acting on the decision to act. CONCLUSION: Although the results should be replicated in a larger and more representative sample, the PAPM seems a good framework for studying fruit intake. PMID- 18421894 TI - Using hypothetical data to assess the effect of numerical format and context on the perception of coronary heart disease risk. AB - PURPOSE: To test the hypothesis that responses to coronary heart disease (CHD) risk estimates are heightened by use of ratio formats, peer group risk information, and long time frames. DESIGN: Cross-sectional, experimental, between factors design. SETTING: Three regions in England. SUBJECTS: A total of 740 men and women ages 30 to 70 years. MEASURES: Risk perception, "emotional" response, intention to change lifestyle. ANALYSIS: Logistic regression was used to investigate the impact of numerical format (ratio vs. percentage), peer group risk (personal vs. peer group), and time frame (10-year vs. 30-year) on risk perception. Analysis of variance was used to investigate the impact of these factors on emotional response and intention to change lifestyle questions. RESULTS: Higher perceived risk was observed when risk was presented as a ratio (p < .001) and when it was supplemented with peer group risk estimates (p = .006). Emotional responses to risk information were heightened when risk was presented as a ratio (p = .0004) and supplemented with peer group risk estimates (p = .002). Presentation with ratios also increased intention to make lifestyle changes (p = .047). CONCLUSION: Perception of CHD risk information is affected by the presentation format. Where absolute risks may appear low, use of ratios and supplementation of personal risk estimates with peer group risk may increase risk perception. PMID- 18421895 TI - Working together to create supportive environments in worksite health promotion. AB - A conccptual model for addressing the creation of supportive environments for worksite health promotion settings is presented. The authors also discuss the variety of organizational and cultural interventions that are necessary in creating a supportive environment for healthy behavior using a social ecology perspective. The connection to next generation strategies in worksite health promotion is proposed and summaries of the relevant literatures are cited. PMID- 18421896 TI - Looking for disease being a model of human aging. AB - This paper is a part of an introduction to authors' study on systemic laminopathies and their role in human aging. Of special interest is progeria--a type of systemic laminopathy associated usually with mutation 1824 C > T and presenting phenotype of preliminary aging. The authors analyse the differences between the progeria and other syndrome of preliminary aging--Werner's syndrome. PMID- 18421897 TI - Tarui disease and distal glycogenoses: clinical and genetic update. AB - Phosphofructokinase deficiency (Tarui disease) was the first disorder recognized to directly affect glycolysis. Since the discovery of the disease, in 1965, a wide range of biochemical, physiological and molecular studies have greatly contributed to our knowledge concerning not only phosphofructokinase function in normal muscle but also on the general control of glycolysis and glycogen metabolism. Studies on phosphofructokinase deficiency vastly enriched the field of glycogen storage diseases, making a relevant improvement also in the molecular genetic area. So far, more than one hundred patients have been described with prominent clinical symptoms characterized by muscle cramps, exercise intolerance, rhabdomyolysis and myoglobinuria, often associated with haemolytic anaemia and hyperuricaemia. The muscle phosphofructokinase gene is located on chromosome 12 and about 20 mutations have been described. Other glycogenoses have been recognised in the distal part of the glycolytic pathway: these are infrequent but some may induce muscle cramps, exercise intolerance and rhabdomyolysis. Phosphoglycerate Kinase, Phosphoglycerate Mutase, Lactate Dehydrogenase, beta Enolase and Aldolase A deficiencies have been described as distal glycogenoses. From the molecular point of view, the majority of these enzyme deficiencies are sustained by "private" mutations. PMID- 18421899 TI - Cardiac autonomic nervous system in patients with myotonic dystrophy type 1. AB - The purpose of the present study was to evaluate cardiac autonomic nervous system (ANS) in patients with myotonic dystrophy type 1 (DM1). The function of ANS was studied in 20 patients with DM1 and 15 healthy controls. All subjects were investigated by a battery of six cardiovascular autonomic tests and power spectral analysis of heart rate variability (HRV). Only one patient had normal autonomic function. Two (10%) patients had mild, 10 (50%) moderate and 7 (35%) severe autonomic dysfunction. Thirteen (65%) patients had vagal and 4 (20%) sympathetic hyperactivity. Seven (35%) patients had vagal and 15 (75%) sympathetic dysfunction. Eighteen (90%) patients had orthostatic hypotension. The 24-hour time domain parameters of SDNN (SD of the NN interval) and total power were significantly lower in DM1 patients than in healthy controls (p < 0.05). However, other parameters of HRV, such as SDANN (SD of the mean NN, 5-minute interval), low frequency (LF), high frequency (HF) power and the LF/HF ratio were somewhat lower in patients with DM1 than in controls, but this was not statistically significant. There was no significant relationship between autonomic dysfunction and the severity of the disease or CTG repeat length. There was also no correlation between HRV and age. Our findings suggest that sympathetic dysfunction and vagal predominance may both occur in patients with DM1. PMID- 18421898 TI - Charcot-Marie-Tooth disease type 4C4 caused by a novel Pro153Leu substitution in the GDAP1 gene. AB - Charcot-Marie-Tooth type 4C4 disease (CMT4C4) is an early onset, autosomal recessive neuropathy with hoarseness caused by mutations in the GDAP1 gene which maps to the 8q13 region. To date, only 24 mutations in the GDAP1 gene have been reported. Neuropathological findings of sural nerve biopsies have been published for a limited number of CMT4C4 patients. Herein, a novel Pro153Leu mutation in the GDAP1 gene identified in a consanguineous Polish family is described and longitudinal clinical and electrophysiological studies as well as morphological findings are presented. PMID- 18421900 TI - Alpha vs. gamma sarcoglycanopathy: DNA tests solve a case from Argentina. AB - Immunohistochemical and DNA results are described in a patient with sarcoglycanopathy. Immunostaining was comparatively normal for alpha-, attenuated for beta- and delta-, and markedly attenuated for gamma-sarcoglycan, thus sarcoglycanopathy was diagnosed, presumably a gamma-sarcoglycanopathy. Unexpectedly, two alpha-SGP-related pathogenic mutations were identified in compound heterozygosity in the SGCA gene: c.229C > T (p.Arg77Cys) in exon 3 and c.850C > T (p.Arg284Cys) in exon 7. These are discussed together with six additional changes detected in SGCB, SGCG and SGCD. PMID- 18421901 TI - [Circulation and heart rate variability at three hemodynamic patterns of juvenile age]. AB - The study involved 135 apparently healthy students 17 - 21 years old. Bioimpedance tetrapolar rheopolygraphy based on Kentavr computer system was employed in studied subjects in supine position at rest. Examined subjects, regarding their percental volume of cardiac index (CI) - 25 and 75, were included into three groups with hypokinetic (CI was lower than 2.0 L/min/m2), eukinetic (CI ranged from 2.0 to 3.0 L/min/m2) and hyperkinetic (CI was more than 3.0 L/min/ m2) hemodynamic pattern. The evaluation of blood flow indices showed that heart rate, stroke volume, minute blood volume, cardiac index, rheogram amplitude were lower in hypokinetic type, but photoplethysmogram was higher when compared with both eukinetic and hyperkinetic types. Power spectral analysis of heart rate variability demonstrated that components of heart rate power frequencies, absolute values of the heart rate power oscillations at four frequencies ranges (UVLF, VLF, LF, HF) as well as relative values of heart rate power spectrum in LF and VLF were lower, but relative values of high-frequency components were higher compared to other two hemodynamic patterns. These data demonstrate that hypokinetic type of circulation was sparer in healthy subjects of juvenile age. PMID- 18421902 TI - [The method of the short term cardiorhytmogram analysis in estimation of estimation of fast change of human activity types]. AB - A new high accuracy method of evaluation of individual RR-intervals distribution is described. The twofold benefits of quantiles measurement of short term RR intervals distribution are high accuracy and reability of results as well as possibility to compare the results among different time spans. The method offered might be used to measure quick changes of human activity. The differences of individual RR-intervals distribution are revealed correlate to type and effectiveness of the activity during 60 sec periods. PMID- 18421903 TI - [Prospects of use coryneformic bacteria drug to prevent and treat respiratory infections]. AB - This work shows the Codivac efficiency in its aerosol form to treat children sick of influenza and acute respiratory viral infections. Course of therapy with the Codivac drug leads to improve clinical picture and rapid elimination virus from human organism as compared with children undergone traditional therapy. Recovery is accompanied with improving of indices of cell immunity. The article shows the prospects of use Codivac to prevent and treat respiratory infections. PMID- 18421904 TI - [Use of molecular-biological microchip TB-BIOCHIP-2 for detecting of Mycobacterium tuberculosis with multidrug resistance to fluoroquinolones in patients with new detected and chronic tuberculosis]. AB - At present the left-handed "respiratory" quinolones such as moxifloxacin and levofloxacin are the most promising drugs for therapy of multidrug resistant tuberculosis (MDR). Fast and specific diagnostics of sensitivity of M. tuberculosis (MBT) with MDR to this group of drugs is required for timely prescription of adequate chemotherapy and its correction in case of MBT resistance to fluoroquinolones. A new generation of biological microchips - TB BIOCHIP-2 makes possible to detect 9 mutation types in quinolones resistant determination region (QRDR) of gene. About 800 samples from 169 patients in Antituberculosis center were studied. In patients with new detected tuberculosis 23.5% MBT resistant to isoniazid and rifampicin (MDR) and sensitive to fluoroquinolones were revealed. In patients with chronic tuberculosis 65.5% MBT MDR were revealed. Our results were confirmed with detecting ofloxacin resistance on Lowenstein - Jensen. In addition efficiency of TB-BIOCHIP-2 to control drug testing sensitivity of MBT-MDR on fluoroquinolones was confirmed. PMID- 18421905 TI - [Teaching of foundation of intellectual property in the system of postdiploma education]. PMID- 18421906 TI - [The interconnections of molecular mechanisms of hormone actions and their role in pathogenesis of obesity, insulin resistance, and diabetes mellitus]. AB - The various hormones, proteins and other compounds related to developing obesity, insulin resistance and type 2 diabetes are analyzed in the paper. 1) Leptin, ciliary neurutrophic factor, adiponectin, glucagon-like peptide 1, peptide YY, neuromedin S, as well as the protein receptors of these hormones decrease the food consumption, increase the energy turnover, and prevent obesity, insulin resistance, and type 2 diabetes development. The mediators of these hormone and receptor actions are melanocyte stimulating hormone (MSH), corticotropin releasing hormone (CRH), and the others. 2) Ghrelin, endogenose cannabinoides, galanin-like peptide and the mediators of their actions: neuropeptide Y (NPY) and Agouti gene related protein (AGRP) increase the appetite and food consumption. Peroxisome proliferation-activated receptor (PPAR) performs the similar action on food intake. The activation of the first group compound functioning decreases the obesity, increases the energy turnover, facilitates the insulin action and prevents the insulin resistance and type 2 diabetes. Increasing the activities of the second group, as well as, decreasing the actions of the first one of substances induce the opposite effects and facilitate obesity, insulin resistance, and type 2 diabetes developments. The interconnections of the molecular mechanisms of so many hormone actions make the very complicated tusk to study the various endocrine disorders including diabetes mellitus as well. PMID- 18421907 TI - [General properties and functioning principles of lectins in biological systems]. AB - The authors give the literature review with analysis of main conceptions of lectins - their structure and functions according to state-of-the-art science and production requirements. Up-to-date definition of lectins based on last achievements of science and practice represented in the article. PMID- 18421908 TI - [Regenerative cell therapy of the type I diabetes mellitus and its complications]. AB - This review describes up-to-date concepts of the type I diabetes mellitus pathogenesis and its complications. The leading role of the immune system malfunction in pancreatic islet of Langerhans and in vessel wall regeneration impairment is emphasized. It is suggested, that cell therapy of the type I diabetes mellitus with pancreatic islet and bone marrow cell transplantation promote beta-cell regeneration due to normalization of intracellular interaction in these tissues and immune homeostasis in whole organism. PMID- 18421909 TI - [The Hirsch index is a new criterion for evaluation of scientific activity]. AB - A new criterion for evaluation of scientific activity by J. Hirsch has been considered. PMID- 18421910 TI - [Inclusion membrane proteins of Chlamydiaceae]. AB - Inclusion membrane proteins belong to the family of unique chlamydial proteins. Members of this family attract attention of scientists because of the following characteristics: Inc-proteins are localized in the inclusion membrane, these proteins have been found in all chlamydial species, expression of the most part of its genes begins during first hours from the infection of cell culture. Biological functions of Inc-proteins remain unknown, but these proteins are suggested to play a key role in process of the development the chlamydial infection. PMID- 18421911 TI - [To the problem of proteolytic enzyme evolution]. AB - Own and literature data on putative evolution of proteolytic enzymes have been reviewed. Modern principles of peptidase classification based on evaluation of homology of more than 66 thousand gene sequences and similarity of general structural organization of almost 2.5 thousands of known enzymes are considered. The review highlights not only evolutionary interrelationships inside related peptidase families, their possible evolutionary background, but also evolutionary determined differences in certain proteolytic pathway in organisms belonging to different taxons. PMID- 18421912 TI - [The state of cytoskeleton and its links "oxygen-peroxide" effects in some pathologies and apoptosis]. AB - The cytoskeleton elements, especially the system of the microtubules, are responsible for production of cell backbone system, creation the global spatial ordered organization for efficient transport processes. Microtubules are involved into transportation of mitochondria, peroxisomes, microsomes, lyzosomes, Golgi apparatus, vesicular structures, some enzymes, adhesion molecules and, possibly, O2-depoting compounds. During disorganization of microtubules, mitochondria (principal consumers of intracellular O2), lose uninterrupted "address" delivery oxidizing substrates and O2. This may be one of important factors underlying mitochondrial dysfunctions accompanied by a rise and/or intensification of cells hyperoxia and oxidative stress. These impairments are obviously responsible for oxygen-peroxide effects in aging, age-related pathologies, carcinogenesis and apoptosis. The agents, both stabilizing and disorganizating microtubules appear to be apoptogenic for the tumors cells. PMID- 18421913 TI - [Ribosomal repeat in the cell free DNA as a marker for cell death]. AB - We have developed a novel method for in vivo evaluation of cell death in patients with acute and/or chronic heart diseases, which are accompanied by apoptosis or cell necrosis. The method is based on the analysis of cell free DNA (cfDNA) in the blood serum (or plasma). The major parameters assessed in the method include total concentration of serum cfDNA, concentration of serum ribosomal repeat (rDNA), content of rDNA in total cfDNA, as well as factors of cfDNA elimination, such as nuclease activity and anti-DNA antibody. We demonstrated a fivefold increase in the serum cfDNA concentration and a 12-fold enhancement of serum rDNA concentration in patients with acute myocardial infarction compared with healthy individuals. In chronic coronary ischemia the serum cfDNA concentration was similar to that in the disease-free group. However, the content of rDNA in cfDNA was 4.8-fold higher, and the serum rDNA concentration was increased sevenfold. We hypothesize that one reason for accumulation of rDNA within cfDNA might be the previously reported resistance of rDNA to the ds-fragmentation by serum endonucleases. In both acute and chronic coronary disease the nuclease activity in the serum was substantially higher than that in the healthy cohort. Moreover, the titer of anti-DNA antibodies was elevated, with these antibodies being mostly bound to the cfDNA. Thus, the release of rDNA fragments into the blood not only reflects cellular death in the body but also determines the response of the organism to the disease-associated stress. PMID- 18421914 TI - [Breast cancer diagnostics based on extracellular DNA and RNA circulating in blood]. AB - Extracellular DNA and RNA were extracted from blood plasma and cell surface-bound fractions of patients with breast tumors and healthy controls. Frequency of RASSF1A, Cyclin D2 and RARbeta2 methylation was detected using methylation specific PCR in the extracellular DNA, extracted from plasma and cell-surface bound fractions of patient blood. Methylation of at least one of these genes was found in plasma of 13% patients with benign breast fibroadenoma and in 60% of breast cancer patients. Using cell-surface bound DNA as a substrate for PCR have lead to increase of gene methylation detection frequency up to 87% in fibroadenoma and 95% in breast cancer patients without false positive controls. GAPDH, RASSF8, Ki-67 RNA and 18S RNA were quantified using RT-qPCR of the extracellular RNA circulating in blood of patients with breast tumors and healthy controls. The main part of the extracellular RNA was shown to be cell-surface bound. Results show a higher amount of RASSF8, Ki-67 RNA and 18S RNA in plasma and cell-bound fraction of patients with breast cancer compared with patients with benign tumors and healthy controls. The data indicate that the specific RNA quantification in blood plasma is valuable for discrimination between cancer and benign tumors, which can be detected with high sensitivity using analysis of methylated RASSF1A, Cyclin D2 and RARbeta2 genes in extracellular circulating DNA. PMID- 18421915 TI - [Spreading and mechanisms of antibiotic resistance of microorganisms, producing beta-lactamases. Molecular mechanisms of resistance to beta-lactams of Klebsiella spp. strains, isolated in cases of nosocomial infections]. AB - Antibiotic sensivity of nosocomial Klebsiella spp. strains (n = 212), isolated from patients treated in 30 medical centers of 15 various regions of Russia was investigated. The Klebsiella genus was represented by the following species: Klebsiella pneumoniae ss. pneumoniae--182 (85.8%), Klebsiella pneumoniae ss. ozaenae--1 (0.5%), Klebsiella oxytoca--29 (13.7%) isolates. The most active antibacterial agents against the investigated strains were carbapenems (imipenem and meropenem). Among 3rd generation cephalosporine the lowest MICs were observed for ceftazidime/clavulanic acid (MIC50--0.25 microg/ml, MIC90--64 microg/ml) and cefoperazone/sulbactam (MIC50--16 microg/ml, MIC90--64 microg/ml). Beta-lactamase genes (TEM, SHV, CTX) were detected in 42 Klebsiella pneumoniae ss. pneumoniae strains by PCR. Alone or in various combinations TEM type beta-lactamases have been found in 16 (38.1%) isolates, SHV--in 29 (69%), and CTX--in 27 (64.3%). Combinations of 2 different determinants were detected in 23.8% of the isolates, 3--in 26.2%. There were not isolates producing MBL class B among resistant to carbapenems nosocomial Klebsiella spp. strains. PMID- 18421916 TI - [Melatonin influence on free radical homeostasis in rat tissues at thyrotoxicosis]. AB - Experimental thyrotoxicosis in rats is accompanied by the increase of serum alanine aminotransferase (AlA), aspartate aminotransferase (AsA), creatine kinase MB (CK-MB) activities and content of primary products of lipid peroxidation- conjugated dienes--in liver, heart and blood. This suggests impairments in these organs accompanying free radical processes intensification. Administration of melatonin decreased AlA, AsA and CK-MB activities and CD level decreased. Thyrotoxicosis increased catalase activity in liver, heart and blood. Exogenous melatonin decreased specific activity ofcatalase in blood and in heart in comparison with animals subjected to hyperthyroidism. However, some increase of catalase specific activity (approximately 15%) was observed in liver. alpha Tocopherol content, raising in rat tissues in thyrotoxicosis development conditions, decreased after melatonin treatment. Thus, exogenous melatonin is capable to reduce lipid peroxidation intensity at thyrotoxicosis and to act as an adoptogen, regulating free radical homeostasis. PMID- 18421917 TI - [Antidepressant discontinuation syndrome--a problem for the clinician and the patient]. AB - Antidepressant drugs are widely used in the treatment of many psychiatric disorders. The decision to discontinue medication after a successful course of treatment, as well as accidental or planned interruptions to treatment, may, in susceptible patients, result in troublesome symptoms which are generally termed "discontinuation". Discontinuation symptoms are now known to be associated with most classes of antidepressants if medication is discontinued without appropriate down-tapering of dose and/or dose frequency. Symptoms of discontinuation may be mistaken for physical illness, relapse into psychiatric disorder or "addictive" potential of antidepressants. Patients' and their families' psychoeducation is still considered a key factor in the management of antidepressant discontinuation syndrome. Prior to initiating treatment or early in its course, clinicians should educate patients about possibility and nature of discontinuation symptoms, the importance of taking medication consistenly. PMID- 18421918 TI - [Lipid peroxidation and copper-zinc superoxide dismutase activity in patients treated with fluoxetine during the first episode of depression]. AB - In recent years researchers have made a lot of studies to determine the molecular and neurochemical mechanisms which are the basis of depressive disorders. Apoptosis is a cause of the limbic system neuronal cells defect in patients suffering from depression. The antioxidant system is the best known molecular mechanism which protects the cells from apoptosis. This system exists inside and outside of the cells compartments. There is much evidence that antioxidant enzymes keep neuronal cells safe from apoptosis, which is a result of oxidative stress. It also limits the premature ageing of cells. AIM: We tried to give an answer to three questions. 1. Is the activity of cooper-zinc superoxide dismutase (CuZnSOD) and lipid peroxydation level (TBARS) different in patients and the healthy control group? 2. Does the activity of CuZnSOD change due to fluoxetine treatment? 3. What is the difference of TBARS concentration in platelets isolated from patients before and after treatment? METHOD: The study comprised of 32 patients diagnosed with depression. The activity of CuZnSOD in platelets was measured by Misra and Fridovich's method. The thrombocyte concentration of TBARS was measured by Placer and coop. method. RESULTS AND CONCLUSIONS: 1. The activity of CuZnSOD in platelets of depressive patients is lower than in the healthy control group, but the differences are not significant. 2. The activity of CuZnSOD rises after fluoxetine treatment. 3. The concentration of TBARS is higher in patients than in the healthy control group. 4. The intensity of lipid peroxydation is statistically lower after fluoxetine treatment. PMID- 18421919 TI - [Can short-term exposure to extremely low temperatures be used as an adjuvant therapy in the treatment of affective and anxiety disorders?]. AB - AIM: The aim of the research was to assess the effect of whole-body cryotherapy (WBCT) on the symptoms observed in a group of patients suffering from affective and anxiety disorders and their own subjective assessment of life satisfaction. METHOD: The study group was given short-term exposure (120-180 sec.) to temperatures between -110 degrees C and -160 degrees C on each working day for a period of 3 weeks (a total of 15 treatments). Both the study group (n=26) and control group (n=34) were observed at the beginning and the end of this 3 week interval. Standard psychopharmacological treatment was carried out in both groups, independently of whether cryotherapy was used or not. Hamilton's scales of depression and anxiety were used, together with the life satisfaction scale. RESULTS: A statistically significant larger improvement, together with a better mean state after 3 weeks, was observed with respect to 11 of the 14 components of the anxiety scale in the study group compared to the control group (except symptoms associated with the gastrointestinal and genitourinary symptoms and behaviour at interview). A larger improvement, together with a better mean state after 3 weeks, was observed with respect to 12 of the 16 components of the depression scale (except digestive, sexual life hypochondria, body weight and criticism) and 6 of the 11 components of the life satisfaction scale (physical well-being, physical condition, domestic activity, professional activity, personal interests and general satisfaction from life) in the study group. CONCLUSIONS: Cyclic short-term whole-body exposition to extremely low temperatures significantly reduced the severity of depressive and anxiety symptoms and increased the life satisfaction. PMID- 18421920 TI - [Evaluation of the activity of selected elements of the immune system in depression]. AB - AIM: The evaluation of the activity of selected elements of the immune system in depression. METHOD: Lymphocyte subsets evaluation (CD3+, CD4+, CD8+, CD16+, CD19+, CD4/CD8) was performed in 32 patients with depression (21 women and 11 men in the age from 21 to 66 years) using the flow cytometry method. The cytokine evaluation (sIL-2R, IL-4, IL-6) was performed in 39 patients with depression (23 women and 16 men in the age from 21 to 66 years) using the ELISA method. The evaluation was also performed in 32 healthy controls (16 women and 16 men in the age from 23 to 61). RESULTS: Statistically significant differences were observed in lymphocyte subsets between depressed patients and healthy controls (increase of CD16+, CD4/CD8, decrease of CD3+, CD8+ in depression, decrease of CD3+, CD8+, CD19+ and increase of CD4/CD8 in remission). There were no significant differences between depressed men and women in the above parameters during exacerbation and remission of depression. Statistically significant differences were observed in cytokine concentration between patients during acute episodes of depressions and healthy controls (higher sIL-2R level, lower IL-4 level), but not in remission. Also, there were no significant differences between men and women in the above parameters during exacerbation and remission of the illness. No correlation was found between age and immunological indicators. CONCLUSION: The results obtained confirm changes of immune system activity in depression, including both activation and supression in the same time. It may suggest an immunological inbalance during depression. PMID- 18421921 TI - [Social networks of depressed patients]. AB - AIM: The purpose of the research was to analyse the structure of social networks as well as types and sources of support received by patients with unipolar disease. METHOD: Assessment was made according to Bizon's method and Cohen's ISEL. RESULTS: In general, the networks of patients function worse than the networks of healthy people. Patients with unipolar disorder do not mention many people of their environment as significant or providing support. They devote less time to social life, are often isolated and lonely and restrict their interactions to family members. That is the group of people from which they obtain most of support, and all supportive functions often lie in one person. These patients have a sense of receiving a small amount of mainly emotional (self esteem) but also appraisal, belonging and tangible support. CONCLUSION: Stabilising supportive relations in different aspects of life should be one of the aims in the therapy both in hospitals and in outpatient departments. Surely that is one of the pathways which may at least to some degree alleviate the huge suffering caused by affective disorders. PMID- 18421922 TI - [Comparison of social networks of patients with unipolar and bipolar disease]. AB - AIM: The purpose of the research was to analyse the structure of social networks as well as types and sources of support received by patients with unipolar and bipolar disease. METHOD: Assessment was made according to Bizon's method and Cohen's ISEL. RESULTS: It has been shown that the functioning of social networks of patients diagnosed as having unipolar affective disorders differs from the ones of bipolar disease. Patients with unipolar disorder do not mention many people of their environment as significant or providing support. They often restrict their interactions to family members. That is the group of people from which they obtain most support. In both groups all supportive functions often lie with one person. Patients with bipolar disorder find it easier to make acquaintance. When compared to the group of bipolar patients, patients with unipolar disease receive less support and the emotional support is the most deficient. CONCLUSIONS: It seems that a combined therapy of affective disorders should include not only pharmacotherapy and various kinds of psychotherapy but also ought to take structure and functioning of patients' social networks into consideration. So stabilising supportive relations in different aspects of life should be one of the aims in the therapy both in hospitals and outpatient departments. PMID- 18421923 TI - [Depression after myocardial infarction and its psychosocial conditions]. AB - Depression after myocardial infarction has influence on patients quality of life, compliance and probably cardiological prognosis. The recognition of factors connected with post infarct depressive symptoms has a significant role for rehabilitation in coronary heart disease. AIM: The aim of the study was: 1) the assessment of depressive symptoms intensity after myocardial infarction (MI), 2) to estimate correlations between post infarct depressive symptoms intensity and present examination of cardiovascular system, past history of depression and coronary heart disease, some psychosocial factors and life events during the one year period before coronary infarct. METHOD: A structured interview--Mini International Neuropsychiatric Interview, Beck Depression Inventory, Social Readjustment Rating Scale and Recent Life Changes Scale were used in the analysis of 102 in-patients after myocardial infarction. RESULTS: Significant correlations between higher depression symptoms intensity after MI and: stressful life events before MI, unemployment, lower education level, history of treated coronary artery disease, depression in the last 6 months before MI, high BMI and benzodiazepine use in the last year were stated. The most often depressive symptoms were not very specific for depressive syndrome. CONCLUSIONS: Clinical (major) depression was diagnosed in 10.8% of the patients after heart infarct. However depressive symptoms (more than 10 Beck points) were present in 40%. The diagnosis of post-myocardial depression may be essentially difficult because of the non-specific character of complains. Significant correlations between intensity of depressive symptoms after myocardial infarct and psychosocial stressors was confirmed. It seems that the most important stressors are those connected with home and family. PMID- 18421924 TI - [Cognitive dysfunctions in patients with alcohol dependence]. AB - AIM: The assessment of cognitive functions connected with prefrontal cortex activity and the intensity of depressed symptoms in patients with alcohol dependence after discontinuation of alcohol use and after a least one year of abstinence. METHOD: 88 patients with alcohol dependence were evaluated. In 51 subjects assessment was performed directly after discontinuation of alcohol drinking and in 37 subjects, after at least one-year of abstinence. The control group consisted in 30 healthy persons matched with age, gender and education for the experimental group. The intensity of alcohol addiction was assessed using the SADD and MAST scale, while intensity of the depression symptoms was measured with the Beck Depression Inventory. Neuropsychological assessment included the Wisconsin Card Sorting Test (WCST) and N-back test for different aspects of working memory and executive functions estimation. RESULTS: Significant disturbances of working memory and executive functions in patients with alcohol dependence, both with short-term and long-term abstinence in relation to healthy subjects were noted. No significant differences between patients with short-term and long-term abstinence on the performance in neuropsychological tests was observed, except better results of nonperseverative errors obtained by patients with long-term abstinence. Longer duration of the alcohol addiction correlated with a worse, however higher level of education with a better performance in neuropsychological tests. The intensity of depression did not correlate with the level of cognitive impairment in the neuropsychological tests. CONCLUSIOS: The results obtained indicate core and enduring cognitive deficits connected with disturbances in prefrontal cortex activity in patients with alcohol addiction. PMID- 18421925 TI - [Detection of alcohol problems among elderly people]. AB - Although alcohol problems among elderly people are common, doctors hardly ever recognize them. There are numerous symptoms observed among people over 65 years old that may signal alcohol problems, but are incorrectly perceived as typical onsets of the ageing process. One of the reasons doctors avoid talking to their patients about alcohol use is the fact they feel incompetent as to the early detection of alcohol use disorders. AIM: The purpose of the study is to present standardized tests aimed at alcohol use disorders detection among elderly people, interpretation of their results and alcohol interview guidelines. DISCUSSION: The alcohol use disorders among elderly people may have serious health consequences. Polish doctors seem to ignore the issue of alcohol use when interviewing both young people and those over 65 years old. The majority of elderly patients see their GPs on regular basis, which is a great opportunity to detect the alcohol problem and then to give proper treatment. The most popular standardized tests aimed at alcohol use disorders detection are: S-MAST-G Test, Quantity/Frequency Screen, CHARM Questionnaire, CAGE Test and its modifications. CONCLUSIONS: Using the presented tests enables preliminary identification of at-risk drinkers, harmful drinkers and alcohol dependents as well as gives an opportunity to recognize people who need a thorough diagnosis or short intervention method. The usage of the presented tools has medical justification because of the fact that drinking alcohol causes serious health consequences and deteriorates the length and quality of life. It is also economical to apply such methods as the cost of screening is low. PMID- 18421926 TI - [Impact of alcohol dependence on the course and psychopathology of schizophrenia]. AB - AIM: The study was undertaken to investigate possible influence of alcoholism on the course and psychopathology of schizophrenia. METHOD: A representative sample of 61 subjects was selected from schizophrenic patients with a history of alcohol dependence (dual diagnosis) registered in the period of 1997-2000. They were compared with 60 control patients with a single diagnosis of schizophrenia. In two groups, with the interview and clinical scales, the onset, course and psychopathology of schizophrenia were assessed. RESULTS: Male patients with schizophrenia and alcohol dependence had a later mean age at onset of mental illness, women--a higher rate of alcohol abuse in the family compared to the control groups. In men with dual diagnosis, the negative symptoms were less prominent. Women with schizophrenia and alcohol dependence reported more depressive symptomatology. PMID- 18421927 TI - [Treatment program for dual-diagnosis substance abusers]. AB - Dual-diagnosis mentally ill patients, i.e. those characterized with substance abuse problems combined with mental health problems, are a challenge both for systems treating substance abusers and for mental health services. These patients are not easily integrated in either of these healthcare systems and/or are treated only for one aspect of their problem by each of these systems. For such patients it is necessary to create a separate treatment model, combining care of the problem of substance abuse and attention to the patient's mental pathology, according to his individual personality traits. For purposes of this programme a treatment setting operating on the model of a therapeutic community is proposed. This setting will open an affiliated treatment programme for dual-diagnosed patients in a separate treatment programme that is not part of the therapeutic community but will be affiliated with it and will accept dual-diagnosis patients. PMID- 18421928 TI - [A review of the effects of nicotine on schizophrenia]. AB - It is increasingly appreciated that amongst psychiatric cigarette smokers, those with schizophrenia have elevated rates of smoking compared to the general population. Nicotine seems to improve cognitive functions critically affected in schizophrenia. There is substantial evidence that nicotine could be used by patients with schizophrenia as a "self-medication" to improve deficits in attention, cognition, and information processing. Perhaps nicotine has influence on intensity of side effects of antipsychotic medication. Nicotine treatment modulates both dopaminergic and glutamatergic neurotransmission, and these effects are specific both to brain region and functional system. Understanding how and why schizophrenic individuals use nicotine may lead to the development of new treatments for both schizophrenia and nicotine dependence. PMID- 18421929 TI - [Occupational medicine as a science for the future]. AB - Paper presents future trends of occupational medicine development, main issues of research and medical education. In paper authors also indicate the crucial role of occupational medicine in the field of health policy, health promotion and prevention. PMID- 18421930 TI - [Risk of cadrdiovascular diseases in lead-exposed workers of crystal glassworks. Part I. Effect of lead on blood pressure and lipid metabolism]. AB - BACKGROUND: Chronic occupational or environmental exposure to small doses of lead results in cardiovascular disorders: arterial hypertension, lipid metabolism disturbances and exacerbation of free radical processes. Each of these changes determines an independent cardiovascular risk factor. The aim of this study was to estimate the cardiovascular risk among the employees exposed to lead in crystal glassworks during the coming 10 years. MATERIALS AND METHODS: A group of 153 persons convinced of their good health status were eligible for the study. A questionnaire was used as a study tool, physical, biochemical and toxicological (free erythrocyte protoporphyrins and lead level in whole blood) studies were performed and the cardiovascular risk factor (SCORE) was individually estimated. RESULTS: Compared to the general population, the increased rate of arterial hypertension, as well as numerous cases of lipid disturbances were revealed. CONCLUSIONS: Workers occupationally exposed to lead should have the cardiovascular risk factor (SCORE) individually estimated. In view of the higher incidence of hypertension in this population, a special attention should be paid to the detection of this pathology and the efficiency of this disease treatment should be controlled. It is necessary to initiate program of individualized actions to decrease so called hard end-points like stroke, myocardial infarct and sudden death. PMID- 18421931 TI - [Attitudes of occupational medicine nurses towards workers' health promotion]. AB - BACKGROUND: The paper outlines the outcomes of a survey aimed at identifying the attitudes of occupational medicine nurses towards health promotion. MATERIAL AND METHODS: The survey was carried out on a random sample of 277 nurses. RESULTS: Almost all respondents think that their occupational group should undertake health promotion activities. However, half of them is convinced that health promotion is only a new name for health education and medical prophylaxis. The vast majority of nurses think that under health promotion programs they should mostly deal with individual health education of patients and encourage them to adopt healthy lifestyles, and they usually undertake this kind of activities. A large number of respondents are not willing to be involved in the organization, marketing, and evaluation of health promotion projects. CONCLUSIONS: There is a great need to intensify measures to motivate nurses to play the roles that are neglected by them, such as looking for new professional groups to undertake activities stimulating health promotion in companies, and developing new institutional and systemic support conducive to making progress in such processes. PMID- 18421932 TI - [Applicability of Voice Handicap Index to the evaluation of voice therapy effectiveness in teachers]. AB - BACKGROUND: The aim of this study was to assess the applicability of Voice Handicap Index (VHI) to the evaluation of effectiveness of functional voice disorders treatment in teachers. MATERIAL AND METHODS: The subjects were 45 female teachers with functional dysphonia who evaluated their voice problems according to the subjective VHI scale before and after phoniatric management. Group I (29 patients) were subjected to vocal training, whereas group II (16 patients) received only voice hygiene instructions. RESULTS: The results demonstrated that differences in the mean VHI score before and after phoniatric treatment were significantly higher in group 1 than in group II (p < 0.05). Moreover, the improvement observed in group I applied to each of VHI subscales: functional, emotional and physical, while in group II it applied only to physical subscale. CONCLUSIONS: This study confirmed that the VHI test can be a useful method for monitoring phoniatric management efficacy in teacher's dysphonia. PMID- 18421933 TI - [Models of workers' health and safety insurance in the selected European Union member states: certain economic problems]. AB - BACKGROUND: The project entitled "An analysis of insurance models in the selected European Union (EU) member States" has recently been accomplished in the Nofer Institute of Occupational Medicine in the section concerning the health and safety of the working population. One of the aims of the project was to identify differences between EU and Polish models, which may provide the basis for a possible involvement of insurance providers (existing and/or created on purpose) in the implementation of tasks in the area of the workers' health protection in Poland. MATERIAL AND METHODS: Documents and publications issued in Poland and elaborated by international organizations were used in the analysis. Of the existing models, those which differ in the solutions concerning the limitation of growing costs of insurance systems, the level of centralization of insurance system management, and the range of cooperation between public and private insurance providers were selected for the analysis. RESULTS: The results of the analysis show that the functioning of insurance systems in the countries under study has been the subject of constant modifications and improvements. Their major aims are to limit the growth of costs of social insurance systems, to shape new qualitative relations between private and public insurance institutions, and to take account of new forms of work regarded as a factor contributing to changes in insurance systems. DISCUSSION AND CONCLUSIONS: The conclusions arising from the analysis of European insurance systems in the area of workplace accidents and occupational diseases, as well as a possible direction of insurance system transformation in Poland address the following issues: the scope of centralization of insurance system management and the role of the state, the degree of independence of insurance institutions and their priority actions for prevention, motivation mechanisms targeted at employers, participation of employers in the consequences of occupational diseases and workplace accidents, as well as the role of additional voluntary workers' health insurance. PMID- 18421934 TI - [Indoor moulds: results of the environmental study in office rooms]. AB - BACKGROUND: The aim of the study was to quantitatively and qualitatively identify moulds occurring in the air of selected office rooms. MATERIAL AND METHODS: The study was conducted in the wintertime inside four office rooms with moisture trace and one control room. To assess mycological contamination of the air, triple samples were directly collected on Petri dishes (Malt Extract Agar medium) using a Burkard air sampler. Qualitative evaluation of moulds was based on the microscopic view and morphological features of colonies. RESULTS: Total number of moulds determined in the air of office rooms did not exceed the level of 4 x 10(2) cfu/m3. The highest concentration of moulds (3.44 x 10(2) cfu/m3) was found in the room that was also used as an archive. It was over tenfold higher than in the control room (0.33 x 10(2) cfu/m3). The qualitative analysis of moulds showed the presence of 15 species. Some of them, including Aspergillus fumigatus, Aspergillus versicolor, Aspergillus ochraceus, Cladosporium spp., Penicillium spp. and Alternaria spp., are recognized as potential health hazards to workers due to the production of mycotoxins and allergic reactions. CONCLUSIONS: The archive was the only room, where concentration of moulds slightly exceeded the reference value. To protect the health of workers who perform their duties in this place, the documents, which are an ideal place of mould growth, should be moved to a separate room. Respiratory complaints reported by workers could be associated with concentrations and species of moulds found in the office rooms under study. PMID- 18421935 TI - [Assessment of carcinogenicity of formaldehyde based on the newest literature data]. AB - In 2004, the category of formaldehyde carcinogenicity was changed in Poland so that it is now coherent with the requirements of the European Union classification of carcinogenicity. Formaldehyde was categorized into group 3 as the substance not classifiable as to its carcinogenecity to humans because the collected information was not adequate for making a satisfactory assessment. There is some evidence from appropriate animal studies, but it is insufficient to categorize this substance in group 2: probably carcinogenic to humans. In 2006, the International Agency for Research on Cancer categorized formaldehyde in group 1 as the substance carcinogenic to humans. Three types of cancer were assessed: nasopharyngeal cancer, leukemia and sinonasal cancer. There was sufficient evidence that formaldehyde causes nasopharyngeal cancer, strong but not sufficient evidence that the substance induces leukemia and limited evidence for sinonasal cancer. Based on the irritation effect, the value of maximum admissible concentration of formaldehyde in the work environment has been established in Poland at 0.5 mg/m3 as average weighed concentration, and 1 mg/m3 as short-term concentration. In the European Union, the values of 0.2 ppm and 0.4 ppm, respectively are now being considered. It is assumed that the MAC value for formaldehyde in the work environment will be soon verified in Poland and adapted to European Union standards. PMID- 18421936 TI - [Medical certification for diabetic drivers in the selected European Union member states]. AB - There is in Poland an urgent need to set criteria and recommendations for medical certification of individuals working or taking up the job involving public safety, in which diabetes could have a particularly important and dangerous impact. In Europe, many countries have restrictions for diabetic drivers. Diabetes is identified as one of the medical conditions that may impair driving ability and thus drivers with diabetes must meet certain medical standards. The European Union legislation states that diabetic patients taking insulin are excluded from driving trucks, heavy goods vehicles, and buses, except for small trucks in "very exceptional cases". This directive is interpreted differently throughout the European Union. Several aspects should be taken into account when evaluating the current status of the medical certification: e.g., duration of diabetes, type of diabetes treatment, diabetes stability, hypoglycemias in the past, and diabetes complications. PMID- 18421937 TI - Laboratory tests for strength paramaters of brain aneurysms. AB - The presentation is focused on the analysis of biophysical properties of cerebral aneurysms, diagnosed and delineated in living human individuals. An aneurysm is a bulging out of a part of the wall of a blood vessel. The aim of our research was to delineate flow patterns inside the aneurysm and its parent artery, to estimate stresses at critical points of the aneurysm wall, to model the haemodynamic effect of different surgical and endovascular tools in order to define the optimal one in a particular case, and to estimate the likelihood of a later aneurysm rupture. For this reason we carried out a lot of different laboratory tests to analyse the mechanical parameters of the aneurysm wall. We made a comparative study of some material models reported in the literature to describe the mechanical response of arteries. These are models for incompressible materials. For this reason we perform uniaxial and biaxial measurements to have appropriate parameters for the models of underlying material. PMID- 18421938 TI - A polyethylene-high proportion hydroxyapatite implant and its investigation in vivo. AB - An implant from hydroxyapatite and polyethylene (HA+PE) composite was investigated for the usability in large bone defects. With this aim, the implants were manufactured in blocks by hot compacting the mixture of 80% HA and 20% PE weight ratio. Powders were machined in a lathe in the dimensions of diaphysis of the radius of the mongrel dogs. Then a defect, 1.5 cm in length, was made in the diaphysis of the radius with an operation performed under general anaesthesia in 16 healthy mongrel dogs. The defects were filled with implant as a block. The dogs were observed radiologically in 15-day intervals and examined clinically in certain intervals. The bone samples were taken out from four dogs for the histopatological examinations at the end of the 2nd, 4th, 6th and 12th months, respectively. Clinical examinations indicated the occurrence of slight lameness in all cases at the first month of experiment, but lameness completely disappeared in a further examination. Progressive resorption and new bone formation began in the implants from the first month, but complete resorption was not observed in any case at the end of 12-month period. SEM and optical microscope examinations revealed fibroblast cell with its clear cytoplasmic extensions and osteoblast cells in endosteum in the inner region. Bone formation increasing and extending to the pores of implant in time and blood vessels with lamellar structure and Haversian system were observed. As a result, it was indicated that HA+PE composite implants could be applied with confidence and are useful in treatment of large bone defects in long bone of dogs. PMID- 18421939 TI - Influence of surface modification on friction coefficient of the titanium elastomer couple. AB - This paper presents the results of a study of the friction coefficient of titanium-elastomer couple. The study was carried out with a view to potential future utilization of its results for constructing retentive elements of implanted prostheses. Changes in the friction force were recorded while removing titanium specimens placed between two silicone counter specimens made of Ufi Gel. The influence of the titanium specimen movement speed in relation that of to the counter specimens and the influence of clamping force on the friction force were assessed. Additionally, the surface roughness of titanium specimens differed; in one case, titanium was coated with polyethylene. The effect of introducing artificial saliva between the cooperating surfaces on the friction force and friction coefficient was analyzed as well. Based on the characteristics recorded, the possibilities of shaping the friction coefficient have been assessed, since it is the friction coefficient that determines effective operation of a friction couple through increasing the titanium specimen roughness. The artificial saliva being introduced between the specimens reduces considerably the friction coefficient through a change of the phenomenon model. An increase in the pressure force for the specimens of high roughness entails a reduction of the friction coefficient. The study carried out allows us to identify the roughness parameters, which in turn will enable obtaining the prescribed retention force for friction/membrane couplings. PMID- 18421940 TI - Biomechanical criterion for selecting cancellous bone screws: arthrodesis in the hindfoot. AB - The aim of the paper was to compare primary biomechanical stability of different arthrodesis screws (7.3 mm AO screw with and without washer, 6.5 mm Herbert screw and 6.5 mm Ideal Compression Screw (I.CO.S)). The work necessary to achieve an adequate compressive force with them was compared to the measurement with the AO screw with washer, because this method is for the time being the most commonly used one and is called the golden standard. Compressive force was measured indirectly, via screw tension measurement, with strain gauges method. From the measurements we calculated the work to reach a limit of 60 N and the ratios corresponding to the value of the golden standard: I.CO.S (35.2%), Herbert (89.0%), AO-screw without washer (116%). The I.CO.S showed superior results. Only in the case of extremely poor bone quality, a clear advantage of I.CO.S could be expected in practice. PMID- 18421941 TI - The application of the cell method in a clinical assessment of bone fracture risk. AB - The aim of this paper is to introduce a new technique for in vivo quantification of bone structure fracture risk. The elastic properties of the architecture obtained from bone digital radiographic images are determined using the cell method. Compression tests are simulated and the results of the numerical analysis are indexed. Such an index is considered to be indicative of the bone structure capabilities. The first clinical validation was carried on at CSMMO, Centro Studi Malattie Metaboliche dell'Osso (Gorizia, Italy), showing that this technique can improve the diagnosis and help a physician in the identification of an actual fracture risk. The examination is not expensive, uses instrumentation that is widely available and therefore could be easily introduced in clinical use as a complement to the current osteoporosis diagnosis methodologies. PMID- 18421942 TI - Musculotendon forces derived by different muscle models. AB - The accuracy, feasibility and sensitivity of several different methods for calculating muscle forces during functional activities in humans were investigated. The upper extremity dynamic system was chosen, where the flexion extension of elbow joint was studied. To counteract the redundant mechanisms we adopted optimization criteria with and without models of individual muscles according to their active and passive properties. Comparisons with known movements solved by inverse dynamics approach and optimization techniques provided similar results for all optimization criteria. Moreover, if muscle models with active and passive properties are included in these analyses, it is relatively easy to calculate muscle forces of both agonists and antagonists. These approaches may be used to provide input data for dynamic FEM stress analysis of bones and bone-implant systems. PMID- 18421943 TI - Experimental and numerical approach to chosen types of mandibular fractures cured by means of miniplate osteosynthesis. AB - The paper presents experimental and numerical analyses of two cases of mandibular corpus fractures cured by means of miniplate implantation. In the laboratory tests, strain gauges and electronic speckle pattern interferometry methods were used, while in the numerical simulations finite element analyses were applied. The aim of such a combined approach was to verify a correctness of the numerical model applied with regard to the assumptions and simplifications which had been done when creating FEM for human mandible: healthy, broken and stabilized with miniplate implants. PMID- 18421944 TI - The evaluation of energy cost of effort and changes of centre of mass (COM) during race walking at starting speed after improving the length of lower extremities. AB - The purpose of the study was to evaluate the influence of extremities length improvement in the form of special shoe orthoses on the walking energy cost of the leading Polish representative in race walking. Before a proper study, the aerobic capacity of the subject was evaluated. The test consisted of two walking efforts performed on a mechanical treadmill. The subject was walking for 12 minutes with shoe orthoses at constant speed (12 km/h) and then the rest phase allowed for the total covering of the oxygen debt. Then the trial was repeated without orthoses. Simultaneously with measuring physiological variables, there was made 3D recording of the athlete's movements on the treadmill applying the Vicon system. There were chosen vertical oscillations of the body center of gravity and work of the subject's system of motion connected with kinetic and potential energy changes regarding the movements of COM during gait. The energy cost of walking at speed related to anaerobic threshold (starting) using shoe orthoses was slightly lower compared to energy expenditure during gait without improvement. No significant differences were noticed in the range of summary vertical COM oscillations during walking in both variants of the measurement. However, considerable asymmetries appeared in the value of COM kinetic energy changes that were lower for the right leg. This testifies to a strongly fixed asymmetrical scheme of individual athlete's technique. PMID- 18421945 TI - Finger cooling test and psychometric analysis in thyroid auxiliary diagnostics. AB - Hypothyroidism is a fairly common disease. The signs and symptoms of thyroid hormone deficiency are non-specific and under certain conditions its diagnosis may be rather difficult. The aim of the study was to make an analytic comparison of several psychometric indices with rheological parameters of extremity microcirculation in patients with subclinical or overt hypothyroidism and in healthy controls. Serum levels of thyrotropin and of free thyroxine were measured. A psychometric analysis was carried out using the Mittenecker and Thoman test. The finger cooling test evaluated by eye verification and the systolic-diastolic index (SDI) was determined by means of planimetric photoplethysmography. An increase in SDI was found in the patients with subclinical and overt hypothyroidism, correlating with psychometric indices and hormonal levels. The results obtained indicate that an evaluation of theological parameters of the finger cooling test and psychometric parameters may be very useful in the auxiliary screening of hypothyroidism. PMID- 18421946 TI - [Response properties of the jaw-closing muscle spindle during decreased occlusal vertical dimension in rats]. AB - The masseter-muscle spindle is regarded as being highly adaptable to increases in the occlusal vertical dimension (iOVD), it is hypothesized that spindle function would adapt to a decrease in occlusal vertical dimension (dOVD) as well. Seventy five 5-week-old female Wistar rats were divided into Control (n = 25) and Experimental (n = 50) groups; those in the Experimental group received a 2.0-mm composite resin build-up to the maxillary molars. The Experimental group was divided into the resin-removal group (n = 25, build-up resin was removed) and the non resin-removal group (n = 25) 8 weeks later; i. e., when the animals were 13 weeks old. Electrophysiological recordings were obtained from masseter-muscle spindle afferents in 13, 14, 15, 19, 21-week-old rats (n = 5 rats each) under general anesthesia Masseter-muscle spindle sensitivity was significantly lower in the resin-removal group 1 week after resin-removal and for the rest of the observation period. The present results indicate that masseter-muscle spindles may not completely adapt to dOVD and may affect jaw function. PMID- 18421947 TI - [Influence of residual stress in wrought wire clasps on bending properties]. AB - The purpose of this study was to assess the influence of residual stress produced after the bending manipulation of a straight wire on flexibility and permanent deformation of wrought-wire clasps. A three-dimensional finite element model of a straight wire (12 mm in length and 1.0 mm in diameter) was created. Elastic and plastic properties of a commercial Co-Cr straight wire measured by means of tensile test were input into the program. We simulated the bending of the clasp shoulder and clasp arm to an angle of 90 degrees or 120 degrees and subsequent unloading processes with spring-back phenomenon. The maximum residual tensile stress appeared at the inside of the bending angle after unloading of the bending force. Thereafter, the clasp models were deflected 0.25, 0.50 mm, and 0.75 mm at the tip to simulate the removal and insertion of a denture. The maximum tensile stress at the inside corner of the bending angle under the deflection was considerably higher than the residual stress before deflection. The stress increase by the deflection was larger at the shoulder than the arm. The load required for deflection was larger in the models with the arm of 120 degrees angle than models with the arm of 90 degrees angle. The results suggest that the permanent deformation of wrought-wire clasps is likely to initiate at the clasp shoulder, while the clasp flexibility is dependent on the bending angle of the clasp arm. PMID- 18421948 TI - [Comparison of gene expression profile of cementoblasts with periodontal ligament cells in mouse mandible with laser capture microdissection]. AB - Cementum is an essential tissue to maintain tooth function and should be closely correlated to tooth root development and periodontal tissue regeneration. However, detailed features of the periodontium including cementum and specific markers for cementoblasts are unknown. Moreover, the molecular mechanism of periodontal tissue development, homeostasis and regeneration remains unknown. Previous studies have usually examined cementum or periodontalligament (PDL) tissue obtained by manual curettage, resulting in difficulties in isolating pure cementum or PDL. We employed laser capture microdissection (LCM) to isolate cementoblasts and PDL cells from undecalcified frozen sections of murine mandible and to obtain RNA of good quality for subsequent genetic analysis. Over 500 cementoblasts and PDL cells were separately laser captured under microscopy. A bioanalyzer detected peaks of 18S and 28S rRNA both in the laser-dissected cementoblasts and in PDL cells, suggesting that the RNA was of sufficient quality. The RNA samples were amplified due to their small amount and a comparative analysis of mRNA expression by GeneChip showed that about 2,000 genes were differentially expressed between cementoblasts and PDL cells. Both cementoblast-positive and PDL cell-negative genes were serially analyzed by quantitative RT-PCR using RNA samples obtained from mandibles and femurs. Several genes were expressed at higher levels in the mandible than in the femur, suggesting that some might be cementoblast-specific markers. We established a novel experimental system with which to isolate target tissues from single cells in undecalcified frozen sections and to obtain intact RNA. These methodologies could be useful for further investigation of mineralized tissues and to explore tissue-specific factors. PMID- 18421949 TI - [Tissue regeneration induced by a transgene for bone morphogenetic protein-12 (BMP-12) in a bone defect]. AB - BMP-12, other than BMP-2, 4, and 7, is known to induce the formation of tendon/ligament-like tissues. However, the role of the protein in the process of bone or cartilage healing has not been adequately examined. The purpose of the present study was to examine if a plasmid vector encoding BMP-12 would induce tendon/ ligament formation in a rat bone defect model. A lyophilized atelocollagen pellet with 100 microg of a plasmid vector encoding BMP-12 or BMP-2 was implanted in a bone defect of a rat femoral metaphysis. The samples were retrieved at 2, 4 and 8 weeks post-operation and subjected to histology and real time PCR analysis. BMP-12 transgene indicated prominent fibrogenesis at 2 weeks. Consequently, the tissue was substituted by bone at 8 weeks. The gene expression of collagen type III was up-regulated by BMP-12 gene transfer after 2 weeks, followed by an increase of elastin and Six1 genes after 4 weeks. These genes, which are intimately related to tendon/ligament formation, decreased in expression at 8 weeks in preparation for osteogenesis afterward. On the other hand, the BMP-2 transgene quickly induced the synthesis of bone-related genes such as collagen type I and ALP. It is concluded that BMP-12 can initiate the development of tendon/ligament-like tissue in a rat bone defect at early healing stages. Maintaining such generation of tissue on bone could be useful for modifying periodontal tissue engineering. PMID- 18421950 TI - [Effects of oral streptococci on biofilm formation by cariogenic bacteria in dual species cultures]. AB - The effects of oral commensal streptococci (Streptococcus sanguinis, Streptococcus gordonii, Streptococcus mitis, and Streptococcus salivarius) on biofilm formation by cariogenic mutans streptococci (Streptococcus mutans and Streptococcus sobrinus) were investigated. Cell suspensions were cultured on 96 well microtiter plates coated with or without salivary components (SC), and in flow cell systems coated with SC in tryptic soy broth including 0.25% sucrose without dextrose (TSB). The resultant biofilm formations were stained using safranin or a LIVE/DEAD BacLight Viability Kit, and examined with absorbance at 492 nm or using confocal laser scanning microscopy. Mutans streptococci and S. sanguinis biofilms were formed significantly on the polystyrene surfaces in TSB. Further, in combination cultures, S. sanguinis formed a sufficient biofilm when cultured with S. mutans. However, when S. sanguinis was cultured with S. sobrinus, biofilm formation was slightly inhibited. S. gordonii also inhibited biofilm formation in the culture with S. sobrinus, but not when cultured with S. mutans. S. mitis and S. salivarius collapsed the biofilm morphology and inhibited volume development in some conditions when cultured with S. mutans or S. sobrinus. Biofilm formation by mutans streptococci was challenged and collapsed on the whole by culturing with each of the other oral streptococci. These results indicate that co-culturing of multiple species of mutans streptococci and other oral streptococci has physical effects related to previous attachment and colonization on the surface, as well as biological effects to regulate biofilm formation. PMID- 18421951 TI - [Local application of simvastatin to rat incisor sockets augments bone]. AB - Decrease of the alveolar ridge after tooth extraction causes difficulties in prosthodontic treatment, especially in implant supported prostheses. Statins, which are inhibitors of cholesterol synthesis and therapeutic drugs for hypercholestemia, stimulate BMP-2 expression in osteoblasts. The purpose of the present study was to examine how local application of simvastatin, one of the statins, to the tooth socket after tooth extraction affects alveolar bone. A mixture of lactic acid/glycolic acid copolymer (PLGA), alpha-tricalcium phosphate (alpha-TCP), calcium carbonate was prepared as a carrier for simvastatin. Ninety six male Wistar rats, 10 weeks old, were used. After right mandibular incisor extraction, they were divided into eight groups and tooth sockets were treated differently: no treatment (control), PLGA carrier only, or PLGA containing 0.1, 0.25, 0.5, 1.0, 2.0, and 4.0 mg simvastatin, respectively. They were sacrificed at 4 and 8 weeks. Bone mineral content (BMC) of the alveolar ridge was measured with dual-energy X-ray absorptiometry and histological analyses were performed. All groups treated with simvastatin showed higher BMC compared to the control group and PLGA carrier group at 4 and 8 weeks. Histological analyses revealed that the thickness of cortical bone of the statin group increased compared to the control group at 4 and 8 weeks; however, newly-formed bone in the tooth socket of the statin group was less than in those of the control group and PLGA carrier group. The results of this study show that local application of simvastatin may enhance bone formation and preserve the alveolar ridge after tooth extraction. PMID- 18421952 TI - [Effects of hyperbaric oxygen treatment on healing of maxillary distraction osteogenesis in beagle dogs]. AB - Distraction osteogenesis has been widely used even in the craniofacial region. A long fixation time during the consolidation period, however, is a major clinical disadvantage. Hyperbaric oxygen (HBO) has been used to improve healing in ischemic wounds. We have recently started applying hyperbaric oxygen to cleft palate patients after maxillary distraction, but there is little basic evidence. We hypothesized that hyperbaric oxygen would enhance the healing of distraction osteogenesis in the cleft palate model in dogs. A bony segment including a canine was transported proximally into an artificial bone defect in the left palate. Three dogs were treated with hyperbaric oxygen for 20 days just after the distraction and three other dogs underwent only the distraction process (control group). Blood flow of the canine pulp in the bone segment was monitored using a laser Doppler flowmeter throughout the experiment. All the dogs were sacrificed on day 100, and radiological analysis using peripheral quantitative CT and histomorphometric evaluations were performed. Blood flow in the HBO-treated group recovered to the original level about 30 days faster than in the control group (p<0.05). Cortical bone mineral density was significantly higher at the distraction site in the HBO-treated group than in the control group (p<0.05). The histomorphometric analysis revealed that the newly formed bone area was also larger in the HBO-treated group than in the control group (p<0.05). These results suggest that hyperbaric oxygen treatment could be useful for early removal of the distraction device in distraction osteogenesis. PMID- 18421953 TI - [FoxO3a is essential for the maintenance of hematopoietic stem cell pool]. PMID- 18421954 TI - [Prognosis and molecular pathogenesis of adult T-cell leukemia/lymphoma]. PMID- 18421955 TI - [Role of positron emission tomography in the clinical practices and trials for malignant lymphoma]. PMID- 18421956 TI - [Clinical and molecular pathogenesis of follicular lymphoma]. PMID- 18421957 TI - [Array comparative genomic hybridilyzation analysis for unclassified peripheral T cell lymphoma revealed possible subgrouping]. PMID- 18421958 TI - [Risk-adapted therapeutic stratification of infantile acute lymphocytic leukemia based on MLL-gene rearrangement]. PMID- 18421959 TI - [Therapy-related infertility in hematological diseases]. PMID- 18421960 TI - [Prognosis and therapy of adult acute leukemia with 11q23 abnormalities]. PMID- 18421961 TI - [Comprehensive approach to water electrolyte disorders]. PMID- 18421962 TI - [How to manage disorders of water balance (dysnatremia)]. PMID- 18421963 TI - [Approach to disorders of potassium metabolism]. PMID- 18421964 TI - [Edema and dehydration, and concentration and dehydration]. PMID- 18421965 TI - [Selection of fluid treatment based on pathophysiology]. PMID- 18421966 TI - [Membrane transport proteins: update]. PMID- 18421967 TI - [Efficacy of steroid pulse therapy in patients with IgA nephropathy and impaired renal function]. AB - OBJECTIVE: In the current study, we evaluated the efficacy of methylprednisolone (MP) pulse therapy in IgA nephropathy (IgAN) patients with established renal function impairment. PATIENTS AND METHODS: We retrospectively analyzed the effect of MP pulse therapy in patients with histologically active IgAN (8 males, 12 females) whose estimated glomerular filtration rate was less than 60 mL/min/1.73 m2 (33.4 +/- 13.1, mean +/- SD). The efficacy of the MP pulse therapy was analyzed by calculating the regression coefficients (dL/mg/month) from the slopes of the 1/serum creatinine (Cr), urine protein/creatinine ratio (g/g x Cr), and the estimated interval from the pulse therapy to dialysis (that is, for Cr to reach 8 mg/dL, as calculated from the slope of 1/Cr) or the actual interval. RESULTS: All patients showed improved regression coefficients (-0.0214 +/- 0.00166 vs. 0.00236 +/- 0.00895, 1 year before vs. after treatment, p < 0.01). The severity of proteinuria decreased significantly from a mean urine protein/creatinine ratio of 2.9 +/- 1.7 before therapy to 1.1 +/- 0.8 (p < 0.01) at 6 months and 0.8 +/- 0.7(p < 0.01) at 12 months after therapy. Although 7 patients underwent dialysis, the average interval from pulse therapy to dialysis was prolonged from an estimated interval of 1.1 +/- 0.9 years to an actual interval of 4.1 +/- 3.5 years. Six patients showed positive regression coefficients at the last observation (4.1 +/- 3.0 years after therapy). The remaining 7 patients who had not undergone dialysis also showed prolongation of the estimated interval from pulse therapy to dialysis of 5.9 +/- 5.1 years before pulse therapy to 25.7 +/- 20.6 years at the final observation (4.9 +/- 3.5 years after therapy). No serious side effects were observed in any of the patients. CONCLUSION: MP pulse therapy can slow the progression of renal deterioration in patients with active IgAN, even in those patients in whom renal function impairment has set in. PMID- 18421968 TI - [Chronic renal failure in patients successfully treated with hematopoietic cell transplantation]. AB - Recent development of hematopoietic cell transplantation (HCT) has greatly improved the quality of life in critical patients with hematological malignancies. On the other hand, it is a fact that some HCT survivors suffer from chronic renal failure (CRF). We attempted to examine the clinical characteristics of CRF in patients who were successfully treated with HCT in Japan. A retrospective analysis of 158 long-term survivors receiving HCT at Komagome Hospital was undertaken. CRF was designated as less than 30 mL/min of estimated GFR (eGFR) calculated by the MDRD formula. We statistically analyzed the influences of total body irradiation (TBI), graft versus host diseases (GVHD), renal impairment during HCT, new incidence of hypertension after transplantation, age, and gender on CRF, using the multivariate logistic regression analysis. Twenty-seven patients (17.1%) had CRF. Their mean ages were 33.1 +/- 8.87 years and mean eGFR levels were 20.5 +/- 9.50 mL/min/1.73 m2. Fifteen patients were recipients of TBI (55.6 %). CRF became obvious within one year after BMT in 5 patients (18.5%) and later in 22 patients (81.5%). Seven patients(25.9%) finally reached end-stage renal disease (ESRD) at the time of over 10 years after HCT. Multivariate logistic regression analysis showed that TBI, renal impairment during HCT, and new incidence of hypertension after HCT were significantly associated with CRF. Considering that 12 patients without TBI (44.4%) developed CRF, "renal impairment during HCT", the odds ratio of which was the highest, might be the factor most closely associated with CRF. The clinical course of a representative patient who developed ESRD was described. An increase in ESRD patients receiving HCT should be anticipated and would constitute a new important issue in nephrology. PMID- 18421969 TI - [Changes in the prognosis for survival of patients with end-stage renal disease (ESRD) treated by hemodialysis]. AB - We started dialysis treatment in our institution in 1966, and have improved hemodialysis (HD) treatment through the induction of a biocompatible dialysis membrane, recombinant human erythropoietin, activated vitamin D and purification of the dialysate. We verified improvement of the prognosis for survival of patients with ESRD during this forty-year period, retrospectively. A total of 1,690 patients who began dialysis therapy in our hospital between January 1966 and December 2005 was studied (men: 1,047, women: 643, age: 58.6 +/- 17.4 years. They were divided into four groups (A: patients who started dialysis in the period from 1966 to 1975; n = 280, B: 1976-1985; n = 455, C: 1986-1995; n = 499, D: 1996-2005; n = 456). The mean follow-up period was 8.48 +/- 8.53 years. Of the patients 1,588 were treated with HD, 78 with peritoneal dialysis (PD), and 24 with PD or HD. Age at the initiation of dialysis increased gradually (A: 40.1 +/- 14.2 y-o, B: 53.2 +/- 15.8 y-o, C: 60.0 +/- 16.0 y-o, D: 66.4 +/- 13.8 y-o), and diabetics increased (A: 6.4%, B: 19.5%, C: 25.6%, D: 33.4%). A total of 1,180 patients died; 48.5% of these patients died of cardiovascular disease, 21.3% of infectious disease, and 6.4% of malignancy. Only 13 patients had kidney transplants. With the Cox proportional hazard model for HD cases, age at the initiation of dialysis, gender, cause of renal disease, and the periods were significant predictors of mortality. The relative risk of mortality compared with that in A was reduced progressively: 0.796 in period B (95% confidence interval [CI]: 0.659-0.961, p = 0.0178), 0.505 in period C (95% CI: 0.409-0.623, p < 0.0001), and 0.286 in period D (95% CI: 0.223-0.366, p < 0.0001). CONCLUSIONS: Although the number of high-aged patients or diabetics with ESRD increased in these 40 years, the survival of the patients with ESRD improved. PMID- 18421970 TI - [Case of rhabdomyolysis induced by the approved daily dose of a traditional Chinese medicine]. AB - We report a case of a 67-year-old woman with hypokalemic rhabdomyolysis induced by pseudohyperaldosteronism. The pseudohyperaldosteronism in this case was caused by the administration of a traditional Chinese medicine, which contained 2.0 g of licorice in the approved daily dose. She started to suffer from hypertension and general fatigue after taking the medication, but continued it for two years until admission after an episode of diarrhea and vomiting. On admission, severe hypokalemia (1.6 mEq/L) and increased serum creatinine kinase (8,778 IU/L) was noted. With the findings of a high transtubular potassium concentration gradient (TTKG) in spite of low plasma renin activity and a low plasma aldosterone concentration, we suspected licorice-induced pseudohyperaldosteronism as the cause of her hypokalemic rhabdomyolysis. The Chinese medicine was terminated, and she received appropriate hydration and potassium replacement therapy as judged by the value of TTKG with the result that her serum potassium and creatinine kinase levels were normalized without any more adverse events. Since it was only a low dose of licorice (2.0 g/day) that induced hypokalemic rhabdomyolysis in this case, serum electrolytes should be examined in all cases under the possible consumption of licorice. PMID- 18421971 TI - [Case of emphysematous pyelonephritis with sepsis and disseminated intravascular coagulation]. AB - We present a case of emphysematous pyelonephritis (EPN) with sepsis and disseminated intravascular coagulation (DIC). An 87-year-old man with a history of uncontrolled diabetes mellitus (DM) for more than 25 years was admitted to our hospital for altered mental status and high fever. The initial diagnosis was acute pyelonephritis based on the findings of pyuria and right costovertebral angle knock pain. DIC developed rapidly even though empirical antimicrobial therapy had been started immediately. The abdominal CT revealed the presence of gas in the right renal parenchyma; the definitive diagnosis was EPN. Escherichia coli (E. coli) was identified from both blood and urine cultures. We selected medical conservative therapy with antibiotics because of his advanced age and a history of myocardial infarction three months previously. With only noninvasive therapy and no surgical therapy, his condition improved and he was discharged four months after admission. EPN is a rare, severe gas-forming, necrotizing infection of the renal parenchyma and surrounding areas. Over 90% of the cases occur in DM patients and the most common causative organism is E. coli (60%). The mortality rate with only medical conservative therapy is approximately 20% and transurethral and/or percutaneous drainage or nephrectomy are generally reported to be necessary. To our knowledge, no case with EPN over the age of 84 years has been reported. Although his condition was very severe on admission and long-term antimicrobial therapy was necessary, he was ambulatory at the time of discharge. Herein, we report the pertinent EPN literature and discuss the management of EPN. PMID- 18421972 TI - [Molecular mechanism of brain regeneration and reconstruction of dopaminergic neural network in planarians]. AB - Recently, planarians have received much attention because of their contributions to research on the basic science of stem cell systems, neural regeneration, and regenerative medicine. Planarians can regenerate complete organs, including a well-organized central nervous system (CNS), within about 7 days. This high regenerative capacity is supported by pluripotent stem cells present in the mesenchymal space throughout the body. Interestingly, planarians can regenerate their brain via a molecular mechanism similar to that of mammalian brain development. The regeneration process of the planarian brain can be divided into five steps: (1) anterior blastema formation, (2) brain rudiment formation, (3) brain pattern formation, (4) neural network formation, and (5) functional recovery, with several kinds of genes and molecular cascades acting at each step. Recently, we have identified a planarian tyrosine hydroxylase (TH) gene, a rate limiting enzyme for dopamine (DA) biosynthesis, and produced TH-knockdown planarians by the RNA interference technique. Studies of TH-knockdown planarians showed that DA has an important role of the modification in behavioral movement in planarians. Using monoclonal anti-planarian TH antibody, we also found that dopaminergic neurons are mainly localized in the planarian brain. When the planarian body was amputated, newly generated TH-immunopositive neurons were detected in the anterior region at day 3 of regeneration (i.e., the period of neural network formation), and the TH-immunopositive axonal and dendritic neural network in the CNS was reconstructed during day 5-7 of regeneration. In this article, recent advances in elucidating the molecular mechanism of planarian brain regeneration and dopaminergic neurons are reviewed, and its future prospects for contribution of this system to basic science and medical science research are described. PMID- 18421973 TI - [Adult neurogenesis in physiological and pathological conditions]. AB - Generation of new neurons persists in the two restricted regions of adult brain, the dentate gyrus (DG) of the hippocampus and the subventricular zone (SVZ). Newly generated neurons are functionally integrated into the neuronal circuits, which is involved in regulation of brain plasticity. Endogenous neuronal production in the DG and SVZ is expected to provide a continuous source of new neurons that replace degenerated neurons in the injured brain. Recent studies indicate that adult neurogenesis is modified by various brain insults including stroke, epilepsy and neurodegenerative disorders. While up-regulation of neurogenesis in these situations may partially contributes to restoration and regeneration of damaged neural tissues, inadequate cell differentiation and/or excessive supply of new neurons should disturb existing neural circuits. For development of successful regenerative medicine for injured brain, we need to understand more precise and comprehensive mechanisms regulating adult neurogenesis. PMID- 18421974 TI - [Transcription factor network that regulates neural development]. AB - In the developing nervous system, neural stem cells initially proliferate extensively by symmetric cell division and then give rise to neurons by asymmetric cell division. After production of neurons, neural stem cells finally differentiate into glial cells. These processes are regulated by basic helix-loop helix (bHLH) transcription factors: the repressor-type bHLH factors promote maintenance of neural stem cells and differentiation of glial cells, whereas the activator-type bHLH factors induce production of neurons. There are many subtypes of neurons, and bHLH factors alone are not sufficient but other factors such as homeodomain factors are required for specification of these neuronal subtypes. These transcription factors will be useful for regeneration of the nervous system. PMID- 18421975 TI - [Mechanisms for making neocortical diversity--insights from evolutionary developmental biology]. AB - The cerebral cortex is one of the most intricate structures in the vertebrate brain, and enormous expansion in size and complexity are conspicuous features in the mammalian cortex. Although how the cerebral cortex evolved from ancestor brains still remains elusive, recent advances of developmental neurobiology shed light on the molecular mechanisms underlying cortical development, and also provide novel insights into the origin of the mammalian cerebral cortex. Here I introduce classical concepts and modern theories on the process for making morphological and functional diversity of the cerebral cortex. In this review, we especially focus on possible mechanism that provide 1) morphological diversity of the amniotes' cortex, 2) expansion of the mammalian neocortical areas, and 3) advancement of human cognitive abilities, based on accumulating knowledge of the comparative brain anatomy, molecular neurobiology, anthropology and cognitive archaeology. PMID- 18421976 TI - [Programmed cell death in the nervous system]. AB - Programmed cell death (PCD) or apoptosis has crucial roles in the development of the nervous system and in tissue homeostasis. It is generally assumed that about half of the neurons are lost before completion of the neural architecture during development. Neurotrophic cell death in innervating neurons matches the number of neurons to the size of their target field. Damaged and potentially harmful cells are induced to apoptosis. The loss of developing neural cells is involved in the morphogenetic sculpting of the early nervous system and the creation of sexually dimorphic structures. PCD also eliminates pioneer neurons that serve transient functions, such as those that navigate in an environment devoid of other axons. Dysregulation of cell death programmes can feature in developmental and neoplastic disorders of the nervous system, including various neurodegenerative diseases. PCD is ultimately difficult process to measure because of their rapidly degradation by neighbors in a few hours or less. The molecular basis underlying why only certain cells are selected to die at adequate stage and how PCD is integrated with other biological processes are largely unknown. However, recent genetic studies in Drosophila and mice provide new insights into the regulatory system of cell death and survival pathway in vivo. This review describes the current knowledge about the significance and mechanisms of selective cell death in the nervous system. PMID- 18421977 TI - [Molecular basis for establishment and maintenance of dendritic trees]. AB - The human brain receives, processes, stores and transmits complex information with great fidelity. The neuronal network that underlies these functions is comprised of an estimated 10(11) neurons linked by 10(14) synaptic connections between two structurally and functionally different neurites, axons and dendrites. Precise patterning of dendrites as well as axons is essential for correct wiring and function of neural circuits. Given that dendrites are a fundamental determinant of neural wiring, it is not surprising that dendrite defects are associated with a variety of human diseases such as Down's syndrome, Rett syndrome, and Fragile-X syndrome. In this review, I summarize our current understanding of cellular and molecular mechanisms that regulate dendritic field formation and influence the shaping of dendritic arbors. In particular, I attempt to address (1) how neurons take the polarity, (2) how the individual dendritic branches are developed, (3) how the size, shape, and complexity of dendritic arbors are regulated, and (4) following establishment of dendritic fields, how arbors are maintained and in some case how arbors change over time. PMID- 18421978 TI - [Role of Pax6 in the developing central nervous system]. AB - Brain is the most complex organ and believed to contain approximately 10 billion neurons supported by 100 billion glial cells. Due to the recent progress in molecular developmental biology and molecular genetics, the molecular mechanism of operation in controlling the development of central nervous system has been identified and responsible genes have been documented for various deficiencies. The Pax6, a member of the Pax family of transcription factors, is first identified as a master control gene for development of the eyes. We now know that the Pax6 has crucial roles in various developmental processes of the telencephalon including dorsal-ventral and anterior-posterior patterning, specification of neuronal subtypes, neuronal migration and axonal projection. Pax6 deficient mice lack the eyes and nose, and show major defects in cortical lamination and brain size. Recent studies have identified that Pax6 contributes to neurogenesis (production of nerve cells) in the postnatal brain. In this review, we introduce multiple functions of Pax6 in the developing central nervous system and in postnatal neurogenesis. PMID- 18421979 TI - [Molecular mechanism of lissencephaly--how LIS1 and NDEL1 regulate cytoplasmic dynein?]. AB - Lissencephaly is a devastating neurological disorder characterized by smooth cerebral surface, thick cortex and dilated lateral ventricules due to defective neuronal migration. Lis1 was identified as a mutated gene in classical lissencephaly patients, and turned out to be a beta-subunit of platelet activating factor acetylhydrolase. Studies in model organisms, particularly Aspergillus nidulans, as well as those in the mouse, have uncovered an evolutionarily conserved pathway that involves LIS1 and cytoplasmic dynein. LIS1 was subsequently found to associate physically with two NudE orthologs, Ndel and its isoform Ndell. These observations also strengthen that LIS1 gene has been implicated in regulating cytoplasmic dynein. Lis1+/- neurons displayed increased and more variable separation between the nucleus and the preceding centrosome during migration. Dynein inhibition resulted in similar defects in both nucleus centrosome (N-C) coupling and neuronal migration, suggesting that defects in this coupling may contribute to migration defects in lissencephaly. Recent report suggests that LIS1 also plays an important role on the determination cleavage plane of neuronal progenitor cells. Controlled gene deletion of Lis1 in vivo in neuroepithelial stem cells, where cleavage is uniformly vertical and symmetrical, provokes rapid apoptosis of those cells. Ndell is also involved in the regulation of microtubule organization, and becomes the target of various kinases and phosphatases, including CDK5/CDK1, Aurora-A and PP4. Coordinated regulation of cytoplasmic dynein and microtubule organization is vital for proper cell division and cell positioning, which is an important research problem for understanding of corticogenesis and promoting the development of new therapies for lissencephaly and related disorders. PMID- 18421980 TI - [Control of neural cell migration during the development of the central nervous system]. AB - During the embryonic development, neurons migrate from their origin to their final position. Control of neuronal migrations is critical for formation of the complex architecture of the central nervous system. In the developing central nervous system, there are two major patterns of neuronal migrations. One is radial migration and the other is tangential migration. In radial migration, migrating cells move along radial fibers that are processes of radial glial cells. Contact between radial glial cells and migrating cells has been supposed to control radial migration. In tangential migration, some tracts of neuronal migrations are controlled by chemokines. Neuronal migration utilizes its own migratory strategies, such as cell-cell contact with radial glial cells as well as common principles of cell migration like chemoattractants. As cell migrations reflect cytoskeletal changes in the cells, external cues like chemoattractants and cell adhesion finally influence the structure of cytoskeleton in migrating neurons. We summarize previous studies on cell migration and discuss specific mechanisms of neural migration in this review. PMID- 18421981 TI - [Mechanisms of cerebral cortical development]. AB - The development of cerebral cortex is regulated by complex mechanisms. Various classes of neurons migrate from the proliferative zones to the cortex according to a spatiotemporal manner. There are two distinct modes of neuronal migration: radial migration and tangential migration. In the benefit of the advancement of research techniques such as in utero electroporation and time-lapse observation of living tissues, more aspects of the cell and molecular mechanisms that control these two migration modes and their roles in the cortical development have been clarified. PMID- 18421982 TI - [Guidance mechanisms in neuronal and axonal migration]. AB - During development, neurons migrate from their birthplace to their appropriate positions and extend their axons toward their proper targets. These events are crucial for the establishment of CNS cytoarchitectures and neural cuircuits essential for brain functions. Recent genetic and biochemical studies have led to the notion that neurons and axons appear to be guided by similar guidance mechanisms and molecules. They appear to be guided by at least four distinct mechanisms: contact attraction, chemoattraction, contact repulsion, and chemorepulsion. These mechanisms are mediated by evolutionarily conserved ligand receptor system. Extracellualr guidance cues, including Netrins, Slits, Semaphorins and Ephrins, act as ligands, while membrane proteins which are expressed in migrating neurons or axons serve as receptors. A certain guidance cue can bind to several distinct receptors and vice versa, leading to elicit diverse responses in migrating neurons or axons with a relatively small number of guidance cues. PMID- 18421983 TI - [Molecular mechanisms of thalamocortical circuit formation]. AB - Thalamocortical (TC) projection is one of the major neural circuitries in the brain. TC projection has characteristic aspects of cortical area and laminar specificities, and provides a suitable model system to investigate the developmental mechanisms of neural circuit formation in the mammalian brain including human beings. Recent studies with genetic, molecular and cellular biological approaches reveal area and lamina-specific gene expressions in the developing cortex, regulation mechanisms of TC axon growth and branching by these molecules, and activity-dependence of the mechanisms. PMID- 18421984 TI - [Molecular mechanisms for the formation of topographic retinotectal projection]. AB - Topographic maps are a fundamental feature of neural networks in the nervous system. Understanding the molecular mechanisms by which topographically ordered neuronal connections are established during development has long been a major challenge in developmental neurobiology. The retinotectal projection of lower vertebrates including birds has been used as a readily accessible model system. In this projection, the temporal (posterior) retina is connected to the rostral (anterior) part of the contralateral optic tectum, the nasal (anterior) retina to the caudal (posterior) tectum, and likewise the dorsal and ventral retina to the ventral (lateral) and dorsal (medial) tectum, respectively. Thus, images received by the retina are precisely projected onto the tectum in a reversed manner. For the formation of topographic maps, molecular gradients in origin and targets are essential. To search for topographic molecules in the embryonic retina, we performed a large-scale screening and successfully identified a variety of molecules with various asymmetrical expression patterns along both axes in the developing retina. Included were many novel molecules with unknown functions, together with known molecules. Through analyses of these molecules, we can now present gene cascades for the retinal patterning and for the establishment of topographic retinotectal projection. In addition, we identified protein tyrosine phosphatase receptor type O (Ptpro) as a specific PTP that regulates Eph receptors. We show that Ptpro controls the sensitivity of retinal axons to ephrins, and thereby plays crucial roles in the topographic projection. PMID- 18421986 TI - [Neurodevelopmental disturbance in the pathogenesis of major mental disorders]. AB - Neurodevelopmental disturbance may underlie the pathogenesis of major mental disorders, including autism and schizophrenia, based on evidence in epidemiology, clinical psychiatry, brain imaging, and neuropathology. This notion is further supported by the fact that many of genetic susceptibility factors for these disorders have key roles in neurodevelopment. Majority of these genetic factors, such as Neuroligins, SHANK3, Neureglin-1, Dysbindin, and Disrupted-in Schizophrenia-1 (DISC1) are associated with "synapse." Therefore, "synapse" is one of the most promising sites of convergence in regard to molecular pathways for these mental conditions. In this review, we will summarize the updates of schizophrenia and autism research, with an emphasis on neurodevelopmental disturbances. PMID- 18421985 TI - [Molecular genetics of lissencephaly and microcephaly]. AB - Genetic malformations of the cerebral cortex are an important cause of neurological disability in children. The genes implicated in these disorders are essential for normal cerebral cortical development. Therefore, identifying these genes and studying their functions will help us further the understanding of the normal biological mechanisms of brain development. Lissencephaly and microcephaly are two groups of disorders that have been intensely studied and several causative genes within each group have been identified. Type I (classical) lissencephaly is characterized by a smooth-appearing brain with a lack or severe reduction of normal gyri. Three of its identified causative genes (LIS1, DCX and TUBA1A) are related to microtubules, which are essential for neuronal migration in the developing cerebral cortex. Microcephaly vera is a form of microcephaly with four responsible genes reported to date. Three of them (ASPM, CENPJ and CDK5RAP2) localize to the mitotic centrosome, and one (MCPH1) is implicated in cell cycle checkpoint regulation and DNA damage response. This suggests that abnormalities of neural progenitor cell division are fundamental to the pathogenesis of microcephaly vera. These genes for microcephaly vera are also suggested to have played a role in evolutionary volume expansion of the human cerebral cortex. These examples show that genetic studies of lissencephaly and microcephaly have been very fruitful in providing novel insights into various aspects of human cerebral cortical development. PMID- 18421987 TI - [Imaging of the brain malformations]. AB - Cortical dysplasia (CD) is a malformation predominantly affected cerebral neocortex, resulting in disorganized brain cytoarchitecture. Normal cortical lamination is disturbed and neurons are abnormally located. Adjacent white matter is often involved. Chronic seizures are a clinical feature of developmental disorders of the brain, including CD and agenesis of the corpus callosum. A morphologically distinct form of CD is focal cortical dysplasia (FCD) characterized by the presence of cytomegalic neurons, seen in specimens resected from patients with partial epilepsy. The abnormal area usually appears normal externally, but may on occasion be represented by a wilder than normal gyrus, while the cut surface shows blurred grey-and white-matter demarcation. MR images of FCD show blurred grey-and white-matter demarcation and T2 elongation in the white matter. In our cases of FCD, no calcifications or mass effects were observed. The lesions of FCD also bear some resemblance to those of a forme fruste of tuberous sclerosis (solitary cortical tuber). The cytoarchitectual abnormalities in cortical tubers are, however, much more extensive and a characteristic feature of cortical tubers is the presence of subpial clusters of giant astrocytes and sheaves of astrocytic processes. CT images of a solitary cortical tuber show abnormal cortex with high density and frequently associated with calcifications and mass effect. An abnormal dilatated and oriented cerebral fissure is often observed around the lesion. White matter bands and blurred grey and white-matter discrimination are occasionally seen in the white matter near the lesion. FCD is a different lesion from a solitary cortical tuber. PMID- 18421988 TI - [Application of psychophysics to neurology]. AB - Although psychophysics has already been used in many neurological evaluations including the visual and hearing tests, the use of psychophysics has been limited to the evaluation of sensory disorders. In this review paper, however, the author introduced recent attempts to apply psychophysics to the evaluation of higher cognitive functions such as perception of scenes and facial expressions. Psychophysics was also used to measure visual hypersensitivity in a patient with migraine. The benefits of the use of psychophysics in neurological and neuropsychological settings would be as follows. (1) We can evaluate higher cognitive functions quantitatively. (2) We can measure performance both above and below the normal range by the same method. (3) We can use the same stimulus and task as other research areas such as neuroscience and neuroimaging, and compare results between research areas. PMID- 18421989 TI - [Atypical intramedullary layer MRI lesion in patient with carcinomatous meningitis]. PMID- 18421990 TI - [One hundred books which built up neurology (16)--Pierre Gratiolet: De la physionomie et des mouvements d'expression (1865) and Duchenne de Boulogne: Mecanisme de la physionomie humaine (1862)]. PMID- 18421991 TI - Interview with Lucian Leape, MD, HFACHE, adjunct professor of health policy, Department of Health Policy and Management, Harvard School of Public Health. Interview by Kyle L Grazier. PMID- 18421992 TI - The uninsured in the U.S. healthcare system. PMID- 18421993 TI - What do consumers want? PMID- 18421994 TI - Chief nursing officer retention and turnover: a crisis brewing? Results of a national survey. AB - Anecdotal evidence suggests growing concerns about chief nursing officer (CNO) dissatisfaction, intent to leave, and turnover. However, little evidence documents the magnitude of the problem or whether CNO turnover requires direct action. This article reports the results from the first phase of a three-phase study examining CNO turnover and retention in U.S. hospitals. CNOs were invited to complete an online survey to gather data about their experiences with turnover and to identify CNO retention issues. Our sample includes responses from 622 CNOs employed in hospitals and healthcare systems across the United States. Approximately 38 percent of the respondents reported having left a CNO position 13 percent within two years before the survey and 25 percent within five years before the survey. Of these, approximately one-quarter had been asked to resign, had been terminated, or had lost their jobs involuntarily. When asked about the context of their departure, a high percentage reported leaving their position to pursue another CNO position (50 percent) or for career advancement (30 percent); approximately 26 percent reported leaving because of conflicts with the chief executive officer. Of great concern is the finding that approximately 62 percent of respondents anticipate making a job change in less than five years, slightly more than one-quarter for retirement. Respondents clearly indicated that CNO turnover is a problem that requires attention. The knowledge gained from this study can be used by healthcare leaders to develop strategies and policies aimed at recruiting and retaining CNOs and easing the transition for CNOs and others in the organization when CNO turnover does PMID- 18421995 TI - Rural hospitals and Spanish-speaking patients with limited English proficiency. AB - Between 1990 and 2000, the Latino population in the United States increased by 61 percent, becoming the largest minority group. Language differences contribute to patient safety and access to healthcare concerns for limited English proficiency (LEP) Latinos. The objectives of this research were to determine the techniques rural hospitals use to accommodate Spanish-speaking LEP patients, to identify strengths and barriers to providing language services, and to describe local approaches to language assistance services. Surveys were mailed to 841 hospitals in 544 rural counties with moderate to high Latino growth rates between 1990 and 2000. A total of 319 rural hospitals responded. Nearly all rural hospitals reported having tools to help patients communicate language needs. The most commonly used tools include brochures, language identification posters, and language identification cards. Strengths were institutional support for language assistance services, staff willingness to use interpreters, and access to telephone language lines. Principal barriers included lack of funding for interpreters, lack of local language training programs, and lack of state agency support. Hospitals that serve counties with higher Latino population growth rates reported higher demand for services compared with those counties with smaller Latino population growth rates. Several innovative approaches were also identified. Various language accommodation resources, tools, and strategies are available for hospitals to help them serve LEP clientele. Hospitals should routinely review their policies and procedures for language assistance services to ensure compliance with federal and Joint Commission standards. PMID- 18421996 TI - Board engagement in quality: findings of a survey of hospital and system leaders. AB - Hospital governing boards assume an important role in improving delivery of quality care in the hospital. More knowledge about the prevalence and impact of particular board activities can help them perform this role more effectively. This study draws from a survey of hospital and system leaders (presidents/chief executive officers [CEOs]) that was conducted in the first six months of 2006 with a total of 562 respondents. The survey contained 27 questions on various aspects of board engagement in quality. More than 80 percent of the responding CEOs indicated that their governing boards establish strategic goals for quality improvement, use quality dashboards to track performance, and follow up on corrective actions related to adverse events. The adoption of other practices was reported less frequently. Only 61 percent of the respondents indicated that their governing boards have a quality committee. The existence of a board quality committee was associated with higher likelihoods of adopting various oversight practices and lower mortality rates for six common medical conditions measured by the Agency for Healthcare Research and Quality's Inpatient Quality Indicators and the State Inpatient Databases. Hospital governing boards appear to be actively engaged in quality oversight, particularly through use of internal data and national benchmarks to monitor the quality performance of their organizations. Having a board quality committee can significantly enhance the board's oversight function. Other potentially useful activities-such as board involvement in setting the agenda for the discussion on quality, inclusion of the quality measures in the CEO's performance evaluation, and improvement of quality literacy of board members-are currently performed infrequently. PMID- 18421997 TI - Two challenges for here and now. Grappling with RAC and POA requirements. PMID- 18421998 TI - Why ICD-10 is worth the trouble. PMID- 18422000 TI - Bridging the translation gap from ICD-9. PMID- 18421999 TI - Brushing up on ICD-10-PCS. PMID- 18422001 TI - ICD-10 preparation checklist. PMID- 18422002 TI - ICD-10 postcards. Canadians, Australians share experiences with ICD-10 implementation. PMID- 18422003 TI - US organizations look ahead. PMID- 18422004 TI - Population health looks forward to the change. PMID- 18422005 TI - The Omaha System. Coded data that describe patient care. PMID- 18422006 TI - A new focus on process and measure. Raising data quality with a standard coding workflow and benchmarks. PMID- 18422007 TI - Managing source system content in the EHR. PMID- 18422008 TI - Teeth for HIPAA in 2008? CMS announces plans for security "assessments". PMID- 18422009 TI - Getting to know e-HIM coding systems. Settings and disciplines offer myriad of documentation systems for EHR use. PMID- 18422010 TI - The role of the cancer registrar. How cancer registrars ensure quality data for public reporting. PMID- 18422011 TI - Collecting root cause to improve coding quality measurement. PMID- 18422012 TI - Key issues in the 2008 OPPS final rule. PMID- 18422013 TI - Coding preventive care services. PMID- 18422014 TI - ICD-10 in the big picture. PMID- 18422015 TI - On loss aversion in capuchin monkeys. AB - Chen, Lakshminarayanan, and Santos (2006) claim to show in three choice experiments that monkeys react rationally to price and wealth shocks, but, when faced with gambles, display hallmark, human-like biases that include loss aversion. We present three experiments with monkeys and humans consistent with a reinterpretation of their data that attributes their results not to loss aversion, but to differences between choice alternatives in delay of reinforcement. PMID- 18422016 TI - Asymmetry of reinforcement and punishment in human choice. AB - The hypothesis that a penny lost is valued more highly than a penny earned was tested in human choice. Five participants clicked a computer mouse under concurrent variable-interval schedules of monetary reinforcement. In the no punishment condition, the schedules arranged monetary gain. In the punishment conditions, a schedule of monetary loss was superimposed on one response alternative. Deviations from generalized matching using the free parameters c (sensitivity to reinforcement) and log k (bias) were compared in the no punishment and punishment conditions. The no-punishment conditions yielded values of log k that approximated zero for all participants, indicating no bias. In the punishment condition, values of log k deviated substantially from zero, revealing a 3-fold bias toward the unpunished alternative. Moreover, the c parameters were substantially smaller in punished conditions. The values for bias and sensitivity under punishment did not change significantly when the measure of net reinforcers (gains minus losses) was applied to the analysis. These results mean that punishment reduced the sensitivity of behavior to reinforcement and biased performance toward the unpunished alternative. We concluded that a single punisher subtracted more value than a single reinforcer added, indicating an asymmetry in the law of effect. PMID- 18422017 TI - Avoidance of timeout from response-independent food: effects of delivery rate and quality. AB - In three experiments, a rat's lever presses could postpone timeouts from food pellets delivered on response-independent schedules. In Experiment 1, the pellets were delivered at variable-time (VT) rates ranging from VT 0.5 to VT 8 min. Experiment 2 replicated the VT 1 min and VT 8 min conditions of Experiment 1 with new subjects. Finally, subjects in Experiment 3 could postpone timeouts from delivery of pellets that differed in quality rather than quantity (unsweetened versus sweetened pellets). In general, response rates and success in avoiding increased as a function of the rate and quality of the pellets. Also, performance efficiency increased as the experiments progressed, that is, the avoidance response occurred later and later in the response-timeout interval. The results support the conclusion that timeout from reinforcement has functional properties similar to those of more commonly studied aversive stimuli (e.g., shock). PMID- 18422018 TI - Determinants of imitation of hand-to-body gestures in 2- and 3-year-old children. AB - Twenty children, ten 2-year-olds and ten 3-year-olds, participated in an AB procedure. In the baseline phase, each child was trained the same four matching relations to criterion under intermittent reinforcement. During the subsequent imitation test, the experimenter modeled a total of 20 target gestures (six trials each) interspersed with intermittently reinforced baseline trials. In each session, target gestures were selected in a pre-randomized sequence from: Set 1- ear touches; Set 2--shoulder touches; Set 3--midarm touches; and Set 4--wrist touches; subjects' responses to targets were not reinforced. In each target set, half the gestures featured in nursery matching games and were termed common targets whereas the remainder, which were topographically similar but did not feature in the games, served as uncommon targets. The children produced significantly more matching responses to common target models than to uncommon ones. Common responses were also produced as mismatches to uncommon target models more often than vice versa. Response accuracy did not improve over trials, suggesting that "parity" did not serve as a conditioned reinforcer. All children showed a strong bias for "mirroring"--responding in the same hemispace as the modeler. The 2-year-olds produced more matching errors than the 3-year-olds and most children showed a bias for responding with their right hands. The strong effects of training environment (nursery matching games) are consistent with a Skinnerian account, but not a cognitive goal theory account, of imitation in young children. PMID- 18422019 TI - Quantification of drug choice with the generalized matching law in rhesus monkeys. AB - The generalized matching law provides precise descriptions of choice, but has not been used to characterize choice between different doses of drugs or different classes of drugs. The current study examined rhesus monkeys' drug self administration choices between identical drug doses, different doses, different drugs (cocaine, remifentanil, and methohexital), and between drug and drug-paired stimuli. The bias parameter of the generalized matching law was used to quantify preference for one reinforcer over another. Choice between identical drug doses yielded undermatching. Choices between 0.3 microg/kg/injection remifentanil and either 0.1 microg/kg/injection remifentanil or saline plus drug-paired stimuli revealed bias for the 0.3 microg/kg/injection dose. Choice was relatively insensitive to differences in random interval schedule value when one reinforcer was replaced with drug-paired stimulus presentations. Bias for 0.3 microg/kg/injection remifentanil over 10 microg/kg/injection cocaine was seen in one subject, and indifference was generally observed between 0.1 microg/kg/injection remifentanil and 56 microg/kg/injection cocaine and between 30 microg/kg/injection cocaine and 320 microg/kg/injection methohexital. These findings suggest the bias parameter may be useful in quantitatively measuring level of preference, which would be an advantage over concurrent FR procedures that often result in exclusive choice. PMID- 18422020 TI - Differential antagonism of cocaine self-administration and cocaine-induced disruptions of learning by haloperidol in rhesus monkeys. AB - Six rhesus monkeys responding under a three-component multiple schedule were administered haloperidol to determine its effects on cocaine self-administration and on cocaine's disruptive effects on the repeated acquisition and performance of response chains. In the absence of haloperidol, 0.0032-0.032 mg/kg/infusion of cocaine increased response rate and the number of infusions in the self administration component when compared to saline administration, whereas 0.1-0.32 mg/kg/infusion decreased response rate and the number of infusions. When compared to saline administration, the two lowest infusion doses of cocaine had little or no effect on responding in the acquisition and performance components; however, higher infusion doses of cocaine dose-dependently decreased response rate in these components. In addition, the higher doses of cocaine also increased the percentage of errors in the acquisition and performance components. Pretreatment with haloperidol (0.0032 or 0.01 mg/kg, i.m.) antagonized the effects of low doses of cocaine on the number of infusions in the self-administration component, whereas only the 0.01-mg/kg dose antagonized the effects of high doses of cocaine on the number of infusions. Neither dose of haloperidol antagonized the rate decreasing effects of cocaine on responding in the acquisition and performance components significantly; the highest dose of haloperidol alone decreased rates of responding in each component. Antagonism of cocaine's error-increasing effects by haloperidol was only evident at one dose of cocaine (0.032 mg/kg/infusion), and was more complete in the performance components than in the acquisition components. Together, these data show the limited suitability of haloperidol for selectively antagonizing cocaine self-administration in the context of a multiple schedule involving transition behavior, and show the lack of uniform antagonism across operant behaviors. PMID- 18422021 TI - A review of domestic dogs' (Canis familiaris) human-like behaviors: or why behavior analysts should stop worrying and love their dogs. AB - Dogs likely were the first animals to be domesticated and as such have shared a common environment with humans for over ten thousand years. Only recently, however, has this species' behavior been subject to scientific scrutiny. Most of this work has been inspired by research in human cognitive psychology and suggests that in many ways dogs are more human-like than any other species, including nonhuman primates. Behavior analysts should add their expertise to the study of dog behavior, both to add objective behavioral analyses of experimental data and to effectively integrate this new knowledge into applied work with dogs. PMID- 18422022 TI - 'No child need have rickets' revisited. PMID- 18422023 TI - Continuous glucose monitoring system in the screening of early glucose derangements in children and adolescents with cystic fibrosis. AB - BACKGROUND: In cystic fibrosis (CF), diabetes mellitus (DM) is associated with progression of pulmonary disease and nutritional impairment. AIM: To compare oral glucose tolerance test (OGTT) and continuous glucose monitoring system (CGMS) in patients with CF with early glucose derangements. PATIENTS AND METHODS: Thirty two patients with CF (5-20 years) with intermediate glucose values > 7.7 mmol/l during OGTT received a CGMS registration. Patients were classified into those with normal glucose tolerance (NGT), impaired glucose tolerance (IGT) and DM, according to glucose values at 120 min of OGTT and during CGMS. Furthermore BMI z scores, forced expiratory volume in 1 second (FEV1%), number of respiratory infections/year, enzyme supplementation, and HbA1c were evaluated. RESULTS: OGTT and CGMS derangements were in agreement in 43.7% of the patients. BMI z-scores, FEV1%, number of respiratory infections/ year, enzyme supplementation, and HbA1c did not differ among the three groups. HbA1c, correlated positively with 120 min OGTT (r = 0.34; p = 0.059), CGMS area (r = 0.35; p = 0.048) and the number of respiratory infections, and negatively with FEV1%. CONCLUSIONS: Intermediate glucose values during OGTT should be considered as a screening test in patients with CF. CGMS can be useful in studying the early occurrence of glucose derangements in selected patients. PMID- 18422024 TI - Ala54Thr polymorphism of the fatty acid-binding protein 2 gene (intestinal-type FABP) is associated with changes in insulin sensitivity in SGA pubertal girls. AB - INTRODUCTION: Associations between FABP2 Ala54Thr polymorphism and increased fasting insulin concentration, fasting fatty acid oxidation and reduced glucose uptake have been identified in several populations. The aim of this study was to evaluate the association of Ala54 Thr polymorphism of the FABP2 gene with insulin sensitivity in pubertal girls born small for gestational age (SGA). RESULTS: The frequency of the Thr54 allele did not differ between AGA and SGA girls (0.52 vs 0.43). Girls born SGA positive for the Ala/Thr polymorphism were older at the beginning of puberty compared to girls born AGA with the Thr54 allele (p < 0.01). These girls had lower whole body insulin sensitivity index (WBISI) (4.1 +/- 1.7 vs 9.2+/-7.4, p < 0.05), higher leptin (17.3 +/- 5.9 vs 12.1 +/- 13.7, p < 0.02), insulin area under the curve (AUC) (64,272 +/- 9,209 vs 27,981 +/- 15,637, p < 0.001), proinsulin (17.3 +/- 5.4 vs 10.9 +/- 3.6, p < 0.01) and insulinogenic index (4.6 +/- 3.0 vs 2.9 +/- 5.9, p < 0.01). Conversely, girls born SGA positive for the Ala/Thr polymorphism were older at the beginning of puberty (ns) compared to girls born SGA positive for the Ala/Ala polymorphism. These girls had higher insulin AUC (64,272 +/- 9,209 vs 33,322 +/-7,533, p < 0.01), insulinogenic index (4.6 +/- 3.0 vs 2.5 +/- 3.6, p < 0.01) and lower WBISI (4.1 +/- 1.7 vs 6.3 +/- 1.8, p < 0.05). DISCUSSION: Our results suggest that the Thr54 variant of the FABP2 gene could be associated with a synergic effect in the SGA group regarding higher leptin levels (p < 0.05), lower insulin sensitivity by WBISI (p < 0.05) and higher insulin secretion determined by higher insulinogenic index (p < 0.01), insulin AUC (p < 0.01) and beta-cell stress measured by higher proinsulin (p < 0.05). Our data suggest an involvement of genetic factors in the insulin resistance associated with reduced fetal growth and strengthen the hypothesis that this association could be the consequence of interactions between detrimental factors during fetal life and genetic susceptibility. PMID- 18422025 TI - Body mass index and body composition in relation to sexual maturation. AB - AIM: To assess body mass index. (BMI) in relation to sexual maturation and verify how fat-free mass (FFM) and fat mass (FM) contribute to its increase. METHODS: 1,275 White adolescents (656 girls and 619 boys), aged 9 to 18 years, were evaluated according to Tanner stages. FFM and FM were calculated by triceps and subscapular skinfold measurements and leg-to-leg bioelectrical impedance. BMI-for maturation was distributed into percentiles. Fat-free mass index (FFMI) and body fat mass index (BFMI) were calculated based on BMI 50t" ph percentile values. RESULTS: During maturation, as BMI increases, girls gain FFMI (13.3 kg/m2 at B1 and 15.8 kg/m2 at B5) and BFMI (3.7 kg/m2 at B1 and 5.7 kg/m2 at B5)- proportionally more fat-- while boys gain FFMI (14.3 kg/m2 at G1 and 17.9 kg/m2 at G5) and lose BFMI (4.8 kg/m2 at G1 and 3.2 kg/m2 at G5). CONCLUSION: These data suggest that FFMI and BFMI-for-maturation should be assessed in adolescents monitored for overweight and obesity. PMID- 18422026 TI - Pubertal progression and serum lipid profile in obese children. AB - BACKGROUND: During puberty, sex hormones are associated with decrease in serum lipids in healthy individuals. AIM: To investigate the relationships between pubertal development, serum lipid profile and atherogenic indices in obese children and adolescents. METHODS: Body mass index (BMI), blood pressure, and serum lipids of 340 obese children (182 girls, 158 boys) aged 3.6-17.8 years were measured. Atherogenic index, total cholesterol (TC)/high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein-cholesterol (LDL-C)/HDL-C were calculated as indices of atherogenic risk. RESULTS: Weight (F = 62.078; p < 0.001), BMI (F = 21.083; p < 0.001), systolic blood pressure (SBP) (F = 11.956; p < 0.001) and diastolic blood pressure (DBP) (F = 3.947, p = 0.005) differed significantly between pubertal stages and increased with advance in stages in males. HDL-C (F = 4.586; p = 0.004) differed significantly between pubertal stages and decreased with advancing stages in males. Weight (F = 66.915; p <0.001), BMI (F = 23.101; p <0.001), SBP (F = 10.873; p <0.001), DBP (F = 2.839; p = 0.040) and LDL-C (F = 3.072; p = 0.029) differed significantly between pubertal stages and increased with advancing stages in girls. Triglyceride, TC, LDL-C and SBP were statistically significant determinants for HDL-C according to multivariate linear regression analysis. SBP was significantly affected by chronological age, BMI, glucose, TC and HDL-C according to stepwise linear regression analysis. CONCLUSION: The increasing rate of obesity is associated with an increase in cardiovascular risk factors in parallel with advance in pubertal stages, particularly in boys. PMID- 18422027 TI - Association of Trp64Arg polymorphism of beta3-adrenergic receptor with insulin resistance in Polish children with obesity. AB - AIM: To establish the influence of the Trp64Arg variant of the beta3-adrenergic receptor (Trp64Arg- beta3AR) on body mass index (BMI) and insulin resistance (IR) in obese children. METHODS: BMI, presence of the Trp64Arg mutation, plasma glucose and insulin concentrations during an oral glucose tolerance test (OGTT) and IR were determined in 60 obese and 33 normal weight children. RESULTS: The frequency of Trp64Arg was similar in normal weight and obese children. BMI, glucose and insulin concentrations during an OGTT in children with Trp64Argbeta3AR were not different from those with Trp64Trpbeta3AR. IR was confirmed in 42.8% of children with Trp64Argbeta3AR and in 45.6% of children with Trp64Trpbeta3AR (NS). CONCLUSIONS: 1. The similar frequency of the Trp64Argbeta3AR variant in normal weight and obese children suggests that it is not a susceptibility gene for obesity in Polish children. 2. The presence of the Trp64Argbeta3AR variant does not have an unfavourable influence on BMI, glucose or insulin concentrations during OGTT or on IR frequency in Polish obese children. PMID- 18422028 TI - Thyroid dysfunction in Down's syndrome and screening for hypothyroidism in children and adolescents using capillary TSH measurement. AB - Thyroid dysfunction is more common in individuals with Down's syndrome (DS) than in the general population, whose clinical features can mask the presenting signs and symptoms of hypothyroidism. Biochemical screening is necessary; however, venepuncture may be difficult. AIMS: To assess the prevalence of thyroid dysfunction in children and adolescents with DS and the feasibility of screening for hypothyroidism using capillary dried blood spot thyroid stimulating hormone (TSH) from infancy. METHODS: 394 children (217 boys, 177 girls) were clinically assessed for thyroid dysfunction and 305 children (aged 4 months to 18.9 years) were screened for hypothyroidism by capillary whole blood TSH sample. RESULTS: Thyroid dysfunction was detected in 4.6%, with 50% unscreened since neonatal screening. Parents reported minimal distress by fingerprick screening. CONCLUSION: DS is associated with an increased prevalence of thyroid dysfunction, particularly in preschool children. Biochemical screening is essential and capillary whole blood TSH sampling for hypothyroidism is feasible, less invasive and acceptable. PMID- 18422029 TI - Exaggerated thyroid stimulating hormone secretion in children exposed to the Chernobyl nuclear reactor catastrophe. AB - We present results of a long-term study of the morpho-functional state of the thyroid gland and of the functional capacities of the hypothalamic-hypophyseal system, as shown by thyrotropin releasing hormone stimulation, in different groups of children who suffered from the Chernobyl accident. It was shown that the thyroid gland of the children who were evacuated from the 30-km zone was damaged most severely due to the influence of radioactive iodine (131I). Living on radionuclide-polluted territories in conditions of iodine deficiency has been an additional contributory factor in the development of thyroid gland diseases. Latent functional deficiency of the hypothalamic-hypophyseal system can be one of the reasons leading to oncopathology of the thyroid gland. PMID- 18422030 TI - Gender identity of children and young adults with 5alpha-reductase deficiency. AB - Male pseudohermaphroditism (46,XY DSD) due to 5alpha-reductase deficiency has been recognized for the last few decades. There is scant literature on this entity in India. We compiled data on five patients with this disorder. Four of our five patients were reared as females. Our assessment of these children reveals that they had male gender identity from childhood. Three of the four reared as females chose to change gender role at adolescence, while the fourth is still prepubertal. We conclude that all these patients had male gender identity from early childhood. The parents took note of this only after the appearance of male secondary sexual characteristics at puberty, thereby giving an impression of change in gender identity and gender role. PMID- 18422031 TI - A very rare cause of virilization in childhood: ovarian Leydig cell tumor. AB - Virilization in childhood results from an adrenal and/or ovarian disorder. These children usually present with hyperandrogenism: acne, deepening voice, abnormal hair growth, cliteromegaly. Ovarian Leydig cell tumor is a very rare cause of hyperandrogenism in childhood. They are usually benign and surgical treatment is usually curative if the tumor is limited to the ovaries. We present the youngest case of ovarian Leydig cell tumor reported in the literature who presented with hyperandrogenism that was satisfactorily resolved after resective surgery. Rare causes of virilization should be investigated with special attention paid to tumoral disease in the differential diagnosis. PMID- 18422032 TI - Novel mutation in cytochrome P450c17 causes complete combined 17alpha hydroxylase/17,20-lyase deficiency. AB - BACKGROUND: Cytochrome P450c17 (CYP17) has two principal enzyme activities, 17alpha-hydroxylase and 17,20-lyase, which are required for cortisol and androgen biosynthesis, respectively. Mutations in the gene encoding for CYP17 result in 17alpha-hydroxylase deficiency (17OHD), a rare form of congenital adrenal hyperplasia, a disorder characterized by adrenal insufficiency, hypertension, primary amenorrhea and sexual infantilism. We describe a case of complete combined 17OHD caused by mutations in the CYP17 gene. PATIENT: This study evaluates a 19 year-old Korean female born from a non-consanguineous relationship who presented with primary amenorrhea, hypertension, hyperpigmentation, absent axillary hair and pubic hair, and Tanner I breasts. Laboratory evaluation showed markedly elevated adrenocorticotropin and 11-deoxycorticosterone with suppressed plasma renin, aldosterone, and cortisol, consistent with 17OHD. METHODS: Genomic DNA was isolated from peripheral blood leukocytes. The eight exons of the human CYP17 gene were amplified in four segments by polymerase chain reaction. Amplicons were gel-purified and directly sequenced. RESULTS: The patient was found to be compound heterozygous for mutations in exon 6: a novel mutation R358X (CGA--TGA) and Y329 del/ sub (TAC-->AA). Both alterations introduce premature stop codons prior to the hemebinding cysteine and are predicted to completely inactivate the encoded P450c17 proteins. CONCLUSION: This patient is a compound heterozygote for nonsense mutations in the CYP17 gene, which confirms the diagnosis of 17OHD. PMID- 18422033 TI - An infant with cytomegalovirus-induced subacute thyroiditis. PMID- 18422034 TI - Characterization of particulate matter and gaseous emissions of a C-130H aircraft. AB - The gaseous and nonvolatile particulate matter (PM) emissions of two T56-A-15 turboprop engines of a C-130H aircraft stationed at the 123rd Airlift Wing in the Kentucky Air National Guard were characterized. The emissions campaign supports the Strategic Environmental Research and Development Program (SERDP) project WP 1401 to determine emissions factors from military aircraft. The purpose of the project is to develop a comprehensive emissions measurement program using both conventional and advanced techniques to determine emissions factors of pollutants, and to investigate the spatial and temporal evolutions of the exhaust plumes from fixed and rotating wing military aircraft. Standard practices for the measurement of gaseous emissions from aircraft have been well established; however, there is no certified methodology for the measurement of aircraft PM emissions. In this study, several conventional instruments were used to physically characterize and quantify the PM emissions from the two turboprop engines. Emissions samples were extracted from the engine exit plane and transported to the analytical instrumentation via heated lines. Multiple sampling probes were used to assess the spatial variation and obtain a representative average of the engine emissions. Particle concentrations, size distributions, and mass emissions were measured using commercially available aerosol instruments. Engine smoke numbers were determined using established Society of Automotive Engineers (SAE) practices, and gaseous species were quantified via a Fourier transform infrared-based gas analyzer. The engines were tested at five power settings, from idle to take-off power, to cover a wide range of operating conditions. Average corrected particle numbers (PNs) of (6.4-14.3) x 10(7) particles per cm3 and PN emission indices (EI) from 3.5 x 10(15) to 10.0 x 10(15) particles per kg-fuel were observed. The highest PN EI were observed for the idle power conditions. The mean particle diameter varied between 50 nm at idle to 70 nm at maximum engine power. PM mass EI ranged from 1.6 to 3.5 g/kg-fuel for the conditions tested, which are in agreement with previous T56 engine measurements using other techniques. Additional PM data, smoke numbers, and gaseous emissions will be presented and discussed. PMID- 18422035 TI - Mercury oxidation promoted by a selective catalytic reduction catalyst under simulated Powder River Basin coal combustion conditions. AB - A bench-scale reactor consisting of a natural gas burner and an electrically heated reactor housing a selective catalytic reduction (SCR) catalyst was constructed for studying elemental mercury (Hg(o)) oxidation under SCR conditions. A low sulfur Powder River Basin (PRB) subbituminous coal combustion fly ash was injected into the entrained-flow reactor along with sulfur dioxide (SO2), nitrogen oxides (NOx), hydrogen chloride (HCl), and trace Hg(o). Concentrations of Hg(o) and total mercury (Hg) upstream and downstream of the SCR catalyst were measured using a Hg monitor. The effects of HCl concentration, SCR operating temperature, catalyst space velocity, and feed rate of PRB fly ash on Hg(o) oxidation were evaluated. It was observed that HCl provides the source of chlorine for Hg(o) oxidation under simulated PRB coal-fired SCR conditions. The decrease in Hg mass balance closure across the catalyst with decreasing HCl concentration suggests that transient Hg capture on the SCR catalyst occurred during the short test exposure periods and that the outlet speciation observed may not be representative of steady-state operation at longer exposure times. Increasing the space velocity and operating temperature of the SCR led to less Hg(o) oxidized. Introduction of PRB coal fly ash resulted in slightly decreased outlet oxidized mercury (Hg2+) as a percentage of total inlet Hg and correspondingly resulted in an incremental increase in Hg capture. The injection of ammonia (NH3) for NOx reduction by SCR was found to have a strong effect to decrease Hg oxidation. The observations suggest that Hg(o) oxidation may occur near the exit region of commercial SCR reactors. Passage of flue gas through SCR systems without NH3 injection, such as during the low-ozone season, may also impact Hg speciation and capture in the flue gas. PMID- 18422036 TI - Design approaches for a cycling adsorbent/photocatalyst system for indoor air purification: formaldehyde example. AB - A kinetic model for a cycling adsorbent/photocatalyst combination for formaldehyde removal in indoor air (Chin et al. J. Catalysis 2006, 237, 29-37) was previously developed in our lab, demonstrating agreement with lab-scale batch operation data of other researchers (Shiraishi et al. Chem. Engineer. Sci. 2003, 58, 929-934). Model parameters evaluated included adsorption equilibrium and rate constants for the adsorbent (activated carbon) honeycomb rotor, and catalytic rate constant for pseudo-first-order formaldehyde destruction in the titanium dioxide photoreactor. This paper explores design consequences for this novel system. In particular, the batch parameter values are used to model both adsorbent and photocatalyst behavior for continuous operation in typical residential home challenges. Design variables, including realistic make-up air fraction, adsorbent honeycomb rotation speed, and formaldehyde source emission rate, are considered to evaluate the ability of the system to achieve World Health Organization pollutant guidelines. In all circumstances, the size of the required rotating adsorbent bed and photoreactor for single-stage operation and the resultant formaldehyde concentration in the home are calculated. The ability of how well such a system might be accommodated within the typical dimensions of commercial ventilation ducts is also considered. PMID- 18422037 TI - Estimating the resuspension rate and residence time of indoor particles. AB - Resuspension experiments were performed in a single-family residence. Resuspension by human activity was found to elevate the mass concentration of indoor particulate matter with an aerodynamic diameter less than 10 microm (PM10) an average of 2.5 times as high as the background level. As summarized from 14 experiments, the average estimated PM10 resuspension rate by a person walking on a carpeted floor was (1.4 +/- 0.6) x 10(-4) hr(-1). The estimated residence time for PM in the indoor air following resuspension was less than 2 hr for PM10 and less than 3 hr for 2-microm tracer particles. However, experimental results show that the 2-microm tracer particles stayed in the combined indoor air and surface compartments much longer (>>19 days). Using a two-compartment model to simulate a regular deposition and resuspension cycle by normal human activity (e.g., walking and sitting on furniture), we estimated residence time for 2-microm conservative particulate pollutants to be more than 7 decades without vacuum cleaning, and months if vacuum cleaning was done once per week. This finding supports the observed long residence time of persistent organic pollutants in indoor environments. This study introduces a method to evaluate the particle resuspension rate from semicontinuous concentration data of particulate matter (PM). It reveals that resuspension and subsequent exfiltration does not strongly affect the overall residence time of PM pollutants when compared with surface cleaning. However, resuspension substantially increases PM concentration, and thus increases short-term inhalation exposure to indoor PM pollutants. PMID- 18422038 TI - Investigations of ash fouling with cattle wastes as reburn fuel in a small-scale boiler burner under transient conditions. AB - Fouling behavior under reburn conditions was investigated with cattle wastes (termed as feedlot biomass [FB]) and coal as reburn fuels under a transient condition and short-time operation. A small-scale (30 kW or 100,000 Btu/hr) boiler burner research facility was used for the reburn experiments. The fuels considered for these experiments were natural gas (NG) for the ashless case, pure coal, pure FB, and blends of coal and FB. Two parameters that were used to characterize the ash "fouling" were (1) the overall heat-transfer coefficient (OHTC) when burning NG and solid fuels as reburn fuels, and (2) the combustible loss through ash deposited on the surfaces of heat exchanger tubes and the bottom ash in the ash port. A new methodology is presented for determining ash-fouling behavior under transient conditions. Results on the OHTCs for solid reburn fuels are compared with the OHTCs for NG. It was found that the growth of the layer of ash depositions over longer periods typically lowers OHTC, and the increased concentration of ash in gas phase promotes radiation in high-temperature zones during initial periods while decreasing the heat transfer in low-temperature zones. The ash analyses indicated that the bottom ash in the ash port contained a smaller percentage of combustibles with a higher FB percentage in the fuels, indicating better performance compared with coal because small particles in FB burn faster and the FB has higher volatile matter on a dry ash-free basis promoting more burn out. PMID- 18422039 TI - Development of a regenerable system employing silica-titania composites for the recovery of mercury from end-box exhaust at a chlor-alkali facility. AB - The release of mercury to the environment is of particular concern because of its volatility, persistence, and tendency to bioaccumulate. The recovery of mercury from end-box exhaust at chlor-alkali facilities is important to prevent release into the environment and reduce emissions as required by NESHAP (National Emission Standards for Hazardous Air Pollutants). A pilot-scale photocatalytic reactor packed with silica-titania composite (STC) pellets was tested at a chloralkali facility over a 3-month period. This pilot reactor treated up to 10 ft3/min (ACFM) of end-box exhaust and achieved 95% removal. The pilot reactor was able to maintain excellent removal efficiency even with large fluctuations in influent mercury concentration (400-1600 microg/ft3). The STC pellets were regenerated ex situ by regeneration with hydrochloric acid and performed similarly to virgin STC pellets when returned to service. On the basis of these promising results, two full-scale reactors with in situ regeneration capabilities were installed and operated. After optimization, these reactors performed similarly to the pilot reactor. A cost analysis was performed comparing the treatment costs (i.e., cost per pound of mercury removed) for sulfur-impregnated activated carbon and the STC system. The STC proved to be both technologically and economically feasible for this installation. PMID- 18422040 TI - Emissions tradeoffs among alternative marine fuels: total fuel cycle analysis of residual oil, marine gas oil, and marine diesel oil. AB - Worldwide concerns about sulfur oxide (SOx) emissions from ships are motivating the replacement of marine residual oil (RO) with cleaner, lower-sulfur fuels, such as marine gas oil (MGO) and marine diesel oil (MDO). Vessel operators can use MGO and MDO directly or blended with RO to achieve environmental and economic objectives. Although expected to be much cleaner in terms of criteria pollutants, these fuels require additional energy in the upstream stages of the fuel cycle (i.e., fuel processing and refining), and thus raise questions about the net impacts on greenhouse gas emissions (primarily carbon dioxide [CO2]) because of production and use. This paper applies the Total Energy and Environmental Analysis for Marine Systems (TEAMS) model to conduct a total fuel cycle analysis of RO, MGO, MDO, and associated blends for a typical container ship. MGO and MDO blends achieve significant (70-85%) SOx emissions reductions compared with RO across a range of fuel quality and refining efficiency assumptions. We estimate CO2 increases of less than 1% using best estimates of fuel quality and refinery efficiency parameters and demonstrate how these results vary based on parameter assumptions. Our analysis suggests that product refining efficiency influences the CO2 tradeoff more than differences in the physical and energy parameters of the alternative fuels, suggesting that modest increases in CO2 could be offset by efficiency improvements at some refineries. Our results help resolve conflicting estimates of greenhouse gas tradeoffs associated with fuel switching and other emissions control policies. PMID- 18422041 TI - Effect of meteorological parameters on fine and coarse particulate matter mass concentration in a coal-mining area in Zonguldak, Turkey. AB - In this work, the effect of meteorological parameters and local topography on mass concentrations of fine (PM2.5) and coarse (PM2.5-10) particles and their seasonal behavior was investigated. A total of 236 pairs of samplers were collected using an Anderson Dichotomous sampler between December 2004 and October 2005. The average mass concentrations of PM2.5, PM2.5-10, and particulate matter less than 10 microm in aerodynamic diameter (PM10) were found to be 29.38, 23.85, and 53.23 microg/m3, respectively. The concentrations of PM2.5 and PM10 were found to be higher in heating seasons (December to May) than in summer. The increase of relative humidity, cloudiness, and lower temperature was found to be highly related to the increase of particulate matter (PM) episodic events. During non-rainy days, the episodic events for PM2.5 and PM10 were increased by 30 and 10.7%, respectively. This is a result of the extensive use of fuel during winter for heating purposes and also because of stagnant air masses formed because of low temperature and low wind speed over the study area. PMID- 18422042 TI - Hot filter/impinger and dilution sampling for fine particulate matter characterization from ferrous metal casting processes. AB - A study using two stack-sampling methodologies for collecting particulate matter (PM) emissions was conducted using a hot filter followed by a cold impinger sampling train and a dilution sampler. Samples were collected from ferrous iron metal casting processes that included pouring molten iron into a sand mold containing an organic binder, metal cooling, removal of the sand from the cooled casting (shakeout), and postshakeout cooling. The shakeout process contributed more to PM emissions than the metal pouring and cooling processes. Particulate matter less than 2.5 microm in aerodynamic diameter (PM2.5) mass emissions for the entire casting cycle ranged from 3.4 to 4.7 lb/t of metal for the hot filter/impinger method and from 0.8 to 1.8 lb/t of metal for the dilution method. Most of the difference was due to PM captured by the impingers, much of which was probably dissolved gases rather than condensable vapors. Of the PM fraction captured by the impingers, 96-98% was organic in nature. The impinger PM fraction contributed 32-38% to the total suspended particle mass and caused a factor of 2 4 positive bias for PM2.5 emissions. For the pouring and cooling processes only, the factor increased to over seven times. PMID- 18422043 TI - Impact of downward-mixing ozone on surface ozone accumulation in southern Taiwan. AB - The ozone that initially presents in the previous day's afternoon mixing layer can remain in the nighttime atmosphere and then be carried over to the next morning. Finally, this ozone can be brought to the ground by downward mixing as mixing depth increases during the daytime, thereby increasing surface ozone concentrations. Variation of ozone concentration during each of these periods is investigated in this work. First, ozone concentrations existing in the daily early morning atmosphere at the altitude range of the daily maximum mixing depth (residual ozone concentrations) were measured using tethered ozonesondes on 52 experimental days during 2004-2005 in southern Taiwan. Daily downward-mixing ozone concentrations were calculated by a box model coupling the measured daily residual ozone concentrations and daily mixing depth variations. The ozone concentrations upwind in the previous day's afternoon mixing layer were estimated by the combination of back air trajectory analysis and known previous day's surface ozone distributions. Additionally, the relationship between daily downward-mixing ozone concentration and daily photochemically produced ozone concentration was examined. The latter was calculated by removing the former from daily surface maximum ozone concentration. The measured daily residual ozone concentrations distributed at 12-74 parts per billion (ppb) with an average of 42 +/- 17 ppb are well correlated with the previous upwind ozone concentration (R2 = 0.54-0.65). Approximately 60% of the previous upwind ozone was estimated to be carried over to the next morning and became the observed residual ozone. The daily downward-mixing ozone contributes 48 +/- 18% of the daily surface maximum ozone concentration, indicating that the downward-mixing ozone is as important as daily photochemically produced ozone to daily surface maximum ozone accumulation. The daily downward-mixing ozone is poorly correlated with the daily photochemically produced ozone and contributes significantly to the daily variation of surface maximum ozone concentrations (R2 = 0.19). However, the contribution of downward-mixing ozone to daily ozone variation is not included in most existing statistical models developed for predicting daily ozone variation. Finally, daily surface maximum ozone concentration is positively correlated with daily afternoon mixing depth, attributable to the downward-mixing ozone. PMID- 18422044 TI - Modeling analyses of the effects of changes in nitrogen oxides emissions from the electric power sector on ozone levels in the eastern United States. AB - In this paper, we examine the changes in ambient ozone concentrations simulated by the Community Multiscale Air Quality (CMAQ) model for summer 2002 under three different nitrogen oxides (NOx) emission scenarios. Two emission scenarios represent best estimates of 2002 and 2004 emissions; they allow assessment of the impact of the NOx emissions reductions imposed on the utility sector by the NOx State Implementation Plan (SIP) Call. The third scenario represents a hypothetical rendering of what NOx emissions would have been in 2002 if no emission controls had been imposed on the utility sector. Examination of the modeled median and 95th percentile daily maximum 8-hr average ozone concentrations reveals that median ozone levels estimated for the 2004 emission scenario were less than those modeled for 2002 in the region most affected by the NOx SIP Call. Comparison of the "no-control" with the "2002" scenario revealed that ozone concentrations would have been much higher in much of the eastern United States if the utility sector had not implemented NOx emission controls; exceptions occurred in the immediate vicinity of major point sources where increased NO titration tends to lower ozone levels. PMID- 18422045 TI - Effectively addressing mental health issues in child welfare practice. Introduction. AB - This volume is dedicated to advances in policies, programs, and practices for effectively addressing the mental health issues in child welfare practice, and it reflects CWLA's and the Mental Health Advisory Board's commitment to ensuring children and their families receive effective mental health services that lead to their optimal well-being. PMID- 18422046 TI - Creating more trauma-informed services for children using assessment-focused tools. AB - This article promotes integrating assessment and evidence-based practice in the treatment of traumatized children through a review of two newly developed trauma assessment tools: (1) the Child Welfare Trauma Referral Tool (CWT), and (2) Assessment-Based Treatment for Traumatized Children: A Trauma Assessment Pathway Model (TAP). These tools use pathways and algorithms to increase understanding of individual child trauma victims, and assist professionals working with children to make appropriate referral and treatment decisions within both child welfare and mental health contexts. PMID- 18422047 TI - Mental health assessment of infants in foster care. AB - Infants placed in foster care are at high risk for emotional and behavioral problems. Assessment of their mental health must account for their often-adverse life experiences prior to placement and the involvement of multiple systems that shape their lives in lieu of parents' authority. This article presents practice guidelines for infant mental health evaluations with consideration of legal requirements and the unique issues conferred by foster care. PMID- 18422049 TI - Effectively addressing mental health issues in child welfare practice: the family connection. AB - Nonprofit family-run organizations, such as Tennessee Voices for Children (TVC), are providing leadership in advocating for and delivering services to children and families in need. Utilizing a family-driven approach and a staff partially comprised of parent-professionals, TVC's Nashville Connection and Family Connection programs have strengthened families by providing alternatives to state custody for children and families living with serious emotional or behavioral problems. TVC's Nashville Connection and Family Connection programs did this by coordinating support services, building community bridges, and providing comprehensive in-home services. PMID- 18422048 TI - The influence of family environment on mental health need and service use among vulnerable children. AB - Children in child welfare are especially likely to have unmet mental health needs. The role of family factors in children's use of mental health services was examined in a longitudinal sample of 1075 maltreated or at-risk children. Vulnerable family environment (poor family functioning, low social support, and caregiver psychological distress) is an important predictor of children's mental health needs. It also predicts them not having these needs met. PMID- 18422050 TI - Effectively addressing mental health issues in permanency-focused child welfare practice. AB - The purpose of this paper is two-fold: to highlight to the professional community the ongoing and shifting mental health needs of children exiting the child welfare system into permanent plans, dismissed from public caseloads yet still much in need of services, and, to present a model of one agency's integrated, outpatient intervention with promising preliminary data. The model offers a specialized; effective approach to the field to address the needs of families built by adoption and relative caregiving. The increasing number of adoptions, and relative and near-kin placements, mandate that competent, lifelong services to children and their permanent families correspondingly increase. This practice model poses one example of translating this responsibility into action, so that these children are discharged not just to one permanent family, but also with a plan for permanent competent support. PMID- 18422051 TI - Fostering healthy futures: an innovative preventive intervention for preadolescent youth in out-of-home care. AB - Although we have not yet demonstrated empirically that the FHF program is effective in addressing mental health, behavioral, and academic issues among preadolescent youth placed in foster care, we believe we have a very promising model. We have successfully negotiated many challenges of conducting a RCT within a child welfare setting, in large part because of the collaboration and support of our community partners. The ultimate goal of this research is to develop more efficacious interventions, thereby reducing adverse life-course outcomes and promoting healthy futures, not only for youth in out-of-home care, but for all youth at risk. PMID- 18422052 TI - Reducing transfers of children in family foster care through onsite mental health interventions. AB - This article describes a successful pilot project in New York City that effectively reduced the number of transfers or replacements of children in family foster care through the placement of mental health clinicians onsite at two foster care agencies. PMID- 18422053 TI - Positive behavioral interventions and supports: using strength-based approaches to enhance the culture of care in residential and day treatment education environments. AB - Positive Behavioral Interventions and Supports model, first introduced into public schools, has been extended to alternative settings. This article highlights applying PBIS to day treatment and residential treatment education programs increasingly challenged to serve seriously emotionally disturbed youth whose risk factors have become more complex. The results demonstrate a more positive environment enhancing children's treatment and education along with decreasing numbers of safety holds and need for out-of-classroom supports. PMID- 18422055 TI - [Complications during and after surgical removal of mandibular third molars. Impact of patient related and anatomical factors]. AB - The estimation of possible intra- and postoperative complications for surgical removal of third molars in the mandible poses a frequent dilemma in oral surgery. In the present study, the influence of the patient's age and gender, a reduced mouth opening, and the anatomical position of the tooth in the mandible on intra- and postoperative complications were evaluated. In a total of 120 surgically removed third molars, 9.2% intraoperative complications occurred, mainly bleeding. Factors influencing the risk for intraoperative complications were a male patient, a reduced mouth opening, and distally angulated teeth. Postoperative complications were encountered in 6.7%, mainly dry sockets. For this group, a female gender, a higher age, and distally angulated teeth were identified as risk parameters. As most of the patient- and anatomy-related factors are set parameters when evaluating possible risk factors for third molar surgery, only the timepoint of surgery can be influenced by the surgeon. Regarding the increase in intra- and postoperative complications for third molar removal in higher age groups, the prophylactic third molar surgery between the age of 18 and 25 seems justified. PMID- 18422054 TI - Psychotropic medication management for youth in state care: consent, oversight, and policy considerations. AB - The use of psychotropic medications in youth with emotional disturbances in state custody is increasing and presents unique challenges concerning consent and oversight. We examine various means that state child welfare agencies use to provide consent for and oversight of psychotropic medications for children in state custody and describe benefits of a consent process that provides for expert consultation to the child welfare agency and prescribing clinicians, case specific and systemic oversight of psychotropic medication use, and education for stakeholders. PMID- 18422056 TI - [Hydroxyapatite bone substitute (Ostim) in sinus floor elevation. Maxillary sinus floor augmentation: bone regeneration by means of a nanocrystalline in-phase hydroxyapatite (Ostim)]. AB - The range of bone regeneration materials suitable for maxillar bone augmentation has increased steadily in the past few years and there is now a wide variety of materials being used. In the present case report, we analyzed the state of bone regeneration after sinus floor augmentation using a nanocrystalline in-phase synthetic anorganic hydroxyapatite bone grafting material (Ostim). A 60-year-old female patient underwent maxillary sinus floor elevation and the cavity was filled with Ostim three years before. Actually, she presented herself with loosening of the dental implant at position 17, as a result of parafunction. At the time of the insertion of a second implant at position 17, bone samples were taken by using a trepan drilling device from the previously augmented area. These samples were analyzed histologically to determine the extent of bone remodeling around the deposits of Ostim. We found that the Ostim deposits were surrounded largely by woven bone and, in parts, by lamellar bone and had facilitated osteoconductive bone regeneration. The adjacent implant, at position 16, which beared a crown exposed to proper biting forces without parafunction, showed proper clinical and radiological characteristics of complete and firm integration into the area which was also filled with Ostim three years ago. We conclude that the use of the nanocrystalline hydroxyapatite Ostim with its stable volume properties appears to be suitable for maxillary sinus floor augmentation. Furthermore, we even found osteoconductive bone regeneration under Ostim near the site of the loosened implant. PMID- 18422057 TI - [Nutrition assessment during pregnancy. A new weight chart]. AB - Weight charts and patterns of weight gain for pregnant women in Argentina are scarce. The aims of the study were: 1) to design a new reference weight gain chart to asses the nutritional status of pregnant women using the body mass index (BMI); 2) to estimate weight gain patterns, and 3) to compare it with a proposed reference chart. In 326 pregnant women before 16th week gestation at enrollment during 2001-2002 at the Sarda' Maternity Hospital (Buenos Aires), weight, height, mid arm circumference and skinfold were measured and body mass index was calculated. Mean, standard deviation, coefficient of variation and polynomial percentiles 3rd through 97th were generated for each gestational age between 12th to 42nd weeks. Maximum variability was 18.4% after 28th week, mean preconcepcional BMI was 24.2 +/- 4.5 kg/m2 and 96% of newborns were at term with a mean Ponderal Index of 2.7 +/- 0.2 g/cm3. Trimesters weight increments were 0.600 kg +/- 0.473, 6.476 kg +/- 3.739 and 5.388kg +/- 4.233 for the 1st, 2nd and 3rd, respectively. Total weight gain achieved was 12.46 Kg +/- 3.13. Compared with the reference curve, the new one showed concordance at the 50th percentile with the upper limit of the normal range. The new reference charts of weight gain for pregnant women using maternal weight and BMI may be useful in prenatal care to asses nutritional status during pregnancy. PMID- 18422058 TI - [Accuracy of spirometry in the diagnosis of pulmonary restriction]. AB - A restrictive ventilatory defect is characterized by a decreased total lung capacity (TLC). The objective of this study was to determine the accuracy of spirometry to detect pulmonary restriction in patients with or without airflow obstruction in the spirometry. Five hundred and twenty patients were included. Normal values for lung function were determined by using the 95% confidence interval (CI) with Morris reference equation for spirometry and European Respiratory Society equation for lung volume. Spirometries were considered obstructive when FEV1/FVC ratio was <70% and FEV1 was below 95%CI. In patients without obstruction in the spirometry (n = 357) sensitivity and specificity were 42.2% and 94.3% respectively, negative predictive value (NPP) was 86.6% and positive PV (PPV) was 65.2%. In patients with an obstructive spirometry (n = 66) sensitivity increased to 75.8% but specificity decreased to 65.9%. PPV was only 57.8% and NPV 81.5%. Patients showing obstruction in the spirometry and false positives of a low FVC (n = 22) had similar values of FVC (57.36 +/- 13.45 vs. 58.82 +/- 8.71%, p = 0.6451), FEV1 (44.73 +/- 19.24 vs. 44.0 +/- 13.08%, p = 0.8745) and DLCO (67.50 +/-27.23 vs. 77.00 +/-16.00%, p = 0.1299) than true positives. Residual volume (RV) (125.72 +/- 64. vs. 77.96 +/- 29.98%, p = 0.0011) and RV/ TLC ratio (56.89 +/- 12.82 vs. 38.43 +/- 13.07%, p = <0.0001) were significantly higher. We conclude that a decreased FVC or VC cannot be considered evidence of ventilatory restriction in the presence of airflow obstruction on spirometry. Diagnosis of "mixed defects" by spirometry is inaccurate and should be avoided without the measurement of lung volumes. PMID- 18422059 TI - [Lung biopsy for the diagnosis of interstitial lung disease]. AB - The objective of this study was to determine the morbidity, mortality and diagnostic yield of video assisted thoracoscopy (VATS) and thoracotomy lung biopsy in interstitial lung disease (ILD). Clinical records of 71 patients were retrospectively analyzed. There was no difference in mean hospital stay, intensive care unit stay and duration of chest tube drainage in patients with VATS (n = 52) compared with those undergoing open thoracotomy (n = 17). Complications rate (22.2% vs. 21.0%, p = 1.0000) and operating mortality (9.2 vs. 15.7%, p = 0.2738) were also similar. Overall, complications occurred in 16 patients (22.5%). Thirty-day mortality rate was 11.2% (n = 8). Prevalence of immunosupression (4/8 vs. 9/63, p = 0.0325) was significantly higher in the group of patients who died. No surviving patients had higher values of plasmatic urea (50 +/- 20.1 mg/dl vs. 31.2 +/- 10.3 mg/dl, p = 0.0013) or lower values of preoperative oxygen saturation (SaO2): 82.7 +/- 14.8% vs. 92.8 +/- 3.4%, (p = 0.0009). Eleven patients had an acute illness. Those patients did not show a higher complication rate (4/11 vs. 10/45, p = 0.4390) but mortality was significantly higher (4/11, 36.3% vs. 3/45, 7.1%, p = 0.0223). Biopsy allowed a specific histologic diagnosis in 100% of patients and changed therapy in 66.7%. We conclude that surgical lung biopsy is a safe and useful procedure in patients with ILD. However the higher mortality rate in patients with acute symptoms, immunocompromise, or in respiratory failure must be balanced against potential benefits of altering treatment decisions. PMID- 18422060 TI - [Erythrocyte indexes in hereditary spherocytosis]. AB - Hereditary spherocytosis is a group of heterogenous disorders characterized by variability in its clinical manifestations, membrane protein defects and inheritance. We analysed the sensitivity and specificity of mean corpuscular hemoglobin concentration (MCHC) and red cell distribution width (RDW) in the diagnostic screening of hereditary spherocytosis. Ninety-four patients were compared to equal number of healthy, age-matched children. All indexes were derived from measurements obtained by aperture impedance (Coulter Counter Model JT). In patients with hereditary spherocytosis, MCHC (35.67+/-1.33 g/dl) and RDW (20.60+/-4.5%) were significantly higher than in normal control subjects (MCHC 33.48+/-0.68 g/dl, p: 0.000; RDW 13.22+/-0.9%, p: 0.000). By using a cutoff for the MCHC of 34.5 g/dl and for the RDW of 14.5%, both indexes showed a sensitivity of 81% and a specificity of 98.9%. The combination of the two test is an excellent predictor for the diagnosis of hereditary spherocytosis. PMID- 18422061 TI - [Acute ischemic stroke in patients aged 80 or older]. AB - Young and old age stroke groups have different vascular risk profiles for cerebral ischemic events. The objective of the study was to describe the risk factor profile and stroke subtype in this population of very elderly people. We included patients over 80 years old with diagnosis of ischemic stroke and transient ischemic attack registered between June 2003 and June 2006. We described the demographic data and subtype of ischemic stroke. Of 535 patients with cerebrovascular ischemic events, the final diagnosis was stroke in 366 cases and transient ischemic attack in 169. Of these patients 33.5% were over 80 years old (179). The mean age was 84.4 +/- 4.4 years. The most frequent risk factors were: hypertension 82.7%, dyslipemia 40.2% and atrial fibrillation 24.6%. Stroke subtype was: large artery disease 6%, cardioembolic stroke 19.7%, small artery disease 41.7%, and other causes 0.8%. Among traditional risk factors for stroke in our very elderly patients, the most significant were hypertension and dyslipemia. This agrees with previous epidemiological studies. The high incidence of small artery disease in our patients may be explained by the risk factor profile. PMID- 18422062 TI - [Survey of intestinal parasites among an aboriginal community in Salta]. AB - The prevalence of intestinal parasitoses by protozoans and helminths was determined in an aboriginal community located 6 km from Tartagal, province of Salta, Argentina. The age of the inhabitants studied ranged from 1 to 49 years old. A total of 112 stool samples were collected in sodium acetate-acetic acid- formalin solution (SAF). Each sample was obtained in three different days, alternatively. Ritchie biphasic concentration method and Faust flotation technique were applied. For survey of Enterobius vermicularis, 68 samples were collected during six consecutive days using anal swabs. For the diagnosis of Dientamoeba fragilis, the modified trichrome Gomori-Wheatley staining technique was used. One hundred and six (94.6%) subjects resulted positive for enteric parasites. Forty-four (41.5%) individuals were positive for four or more parasites. The parasites more frequently found were, among the protozoans Blastocystis hominis (58.9%), Entamoeba coli (51.8%), Giardia lamblia (27.7%) and Entamoeba histolytica/E. dispar (24.1%). Among the helminths, the most frequent were hookworms (58.0%), Hymenolepis nana (31.2%) and Strongyloides stercoralis (24.1%). This is the first time that Entamoeba histolytica/E. dispar (24.1%) and Dientamoeba fragilis (2.7%) have been registered in the suburbs of Tartagal. This study reveals how intestinal parasitoses constitute a critical problem of public health in aboriginal communities like the one examined, where the lack of adequate sanitation conditions and unsuitable supply of water coexist. PMID- 18422063 TI - [Adherence with national guidelines in hospitalized patients with community acquired pneumonia. Results of CAPO study in Argentina]. AB - A committee of six scientific organizations from Argentina developed guidelines for the management of patients with community-acquired pneumonia (CAP).The objective of this study was to evaluate the level of adherence with the recommended care suggested by the guidelines in patients hospitalized with CAP in Argentina. Using quality indicators the management of 436 patients who were hospitalized in 12 Argentinean institutions was evaluated. The care given among the following areas was reviewed: need for hospitalization, need for oxygen therapy, empiric antibiotic therapy, switch therapy, hospital discharge, and prevention. The level of adherence was classified as optimal (>90%), intermediate (60% to 90%), and low (<60%).The selection of the empiric antibiotic therapy and the administration of antibiotics between the first 8 hours after arrival had an adherence greater to 80%. A low level of adherence was found in the areas of switch therapy (53%), prevention of CAP with pneumococcal vaccine (51%) and smoking cessation offered (29%). Using quality indicators it is possible to identify specific areas of management in patients with CAP to a low level of adherence with national guidelines. In Argentina interventions to improve switch therapy and prevention of CAP should be developed. PMID- 18422065 TI - [Prolonged cough as initial symptom of temporal arteritis]. AB - Prolonged cough could be a diagnostic and therapeutic dilemma. We present here the case of an elder man who suffered from a dry cough and malaise during four weeks, followed by a fever of unknown origin. After many studies and review of symptoms, a biopsy of the temporal artery confirmed a giant cell arteritis. Cough and malaise could be an early symptom of temporal arteritis in elderly patients. Pain in the outer ear on the lateral decubitus could be another interesting symptom in this disease. PMID- 18422064 TI - [Spinal abscess in a patient with hereditary hemorrhagic telangiectasia]. AB - Rendu-Osler-Weber syndrome is an autosomal dominant disorder characterized by multiple skin and mucosal telangiectasis and multiorgan arteriovenous malformations. Neurological manifestations may occur because of cerebral arteriovenous malformations, intracranial hemorrhage, and most commonly by ischemic stroke and brain abscess secondary to paradoxical embolization in patients with pulmonary arteriovenous malformations. Intramedullary abscess is a rare, unusual condition, in Rendu-Osler-Weber syndrome. We report the case of a 56 years old woman, with a familial history of Rendu-Osler-Weber syndrome, admitted to intensive care with acute quadriplegia and hypoxemia. Our diagnosis was pulmonary arteriovenous malformations and intramedullary abscess. PMID- 18422066 TI - [BK virus nephropathy after renal transplantation. Diagnosis and prognosis by real time PCR]. AB - BK virus nephropathy may lead to kidney transplant failure. BK infection and acute rejection are clinically undistinguishable, therefore diagnosis of these entities is critical to establish the correct treatment. The new molecular methods using PCR and real time PCR have significantly contributed to the rapid and sensitive diagnosis of BK virus. Furthermore, viral load determination in plasma has significantly been associated with BK virus nephropathy. Definite diagnosis of nephropathy requires renal biopsy, although due to the multifocal nature of the disease sensitivity may be less than 100%. BK detection in blood and urine by PCR has contributed to the diagnosis of nephropathy in a more standardized and less invasive way. Recently, quantification of BK virus in plasma has been used for the diagnosis and monitoring of this disease. In the present study, we describe the validation of a real time PCR method for BK virus detection in plasma and urine and its application for diagnosis and monitoring in a renal transplant patient with nephropathy. PMID- 18422067 TI - [Rickettsial spotted fever in the Parana Delta. An emerging disease]. AB - We describe a case of rickettsial spotted fever in the Parana Delta region of Buenos Aires province in Argentina. The patient developed an acute febrile syndrome characterized by myalgias, headache, asthenia and moderate odynophagia, followed by a diffuse macular, papular, and purpuric exanthema. The patient had been bitten recently by a tick on the left preauricular region and an erosive papular lesion was evident at the bite site. An indirect immunofluorescence antibody assay identified antibodies reactive with spotted fever group rickettsiae in the patient's serum. The patient improved rapidly with doxycycline. Several considerations relating to the identity of the rickettsial species and tick vector are discussed, including the possibility that this patient's illness may have been caused by Rickettsia parkeri. PMID- 18422068 TI - [Neuronal isoform of nitric oxide syntase in dense macula]. PMID- 18422069 TI - [Annular image in a bacterial endocarditis]. PMID- 18422070 TI - [Treatment of hematologic neoplasms during pregnancy]. AB - The most common hematological malignancy in pregnant patients is Hodgkin's lymphoma, but other diseases such as chronic and acute leukemia or non Hodgkin's lymphoma have also been reported. In the last decade, new drugs have changed the prognostic of acute promyelocytic leukemia, chronic myeloid leukemia and non Hodgkin's lymphoma. Herein we present updated information on drugs and treatments, new developments, mechanism of action, clinical application, experience on treatment outcomes, adverse effects and teratogenesis, with the objective of orienting hematologists, oncologists and pediatricians. The medical team should offer the most efficient treatment available in order to achieve cure or remission of the disease, and also inform on possible risks for the mother and the fetus, as well as those derived from the delay in treatment application. PMID- 18422071 TI - [Macrophages and arginase induction as a mechanism for parasite escape]. AB - Although there are several immunological mechanisms to eliminate the intracellular pathogens, they have elaborated a variety of strategies to escape of the immune response and to make possible their survival and replication in the host. Some parasites modulate the production of several toxic molecules synthesized by the immune system. Several parasites are highly sensitive to nitric oxide (ON) and their derivatives. ON is produced in macrophages (Mphi) after stimulation with microbial products or cytokines. In the past, Mphi were defined as inflammatory cells (classically activated Mphi), able to produce inflammatory mediators, to act like antigens presenting cells and to kill intracellular pathogens. Nevertheless, activated Mphi involve a more heterogeneous group of cells with different biological markers that can carry out different immunological functions. Alternatively activated Mphi fail to produce ON due to the arginase induction and consequently they have diminished their capacity to kill intracellular pathogens. It has been reported the induction of arginase by different parasites; therefore this mechanism could favor their survival in the host. In our group, we studied the participation of arginase in a model of Trypanosoma cruzi infection and the intracellular signals involved in the replication of this parasite in Mphi. The data obtained from our works would allow the understanding of some mechanisms by which cells can be programmed to favor the establishment of chronic parasitic infections. PMID- 18422072 TI - [Type I DNA topoisomerase from protozoan pathogens as a potential target for anti tumoral drugs]. AB - The intensive use of antiparasitic drugs is the main cause of the emergence of multiresistant parasite strains on those regions where these parasites are endemic. The aetiological agents of the so-called tropical diseases viz. malaria, cryptosporidiosis, sleeping sickness, Chagas disease or leishmaniasis, among others, are unicellular protozoan parasites with no immune-prophylactic treatment and where the chemotherapeutical treatment is still under controversy. At present, the chemotherapeutic approach to these diseases is expensive, has side or toxic effects and it does not provide economic profits to the Pharmaceuticals which then have no or scarce enthusiasm in R & D investments in this field. The identification of type I DNA-topoisomerases as promising drug targets is based on the excellent results obtained with camptothecin derivatives in anticancer therapy. The recent finding of significant structural differences between human type I DNA-topoisomerase and their counterparts in trypanosomatids has open a new field in drug discovery, the aim is to find structural insights to be targeted by new drugs. This review is an update of DNA-topoisomerases as potential chemotherapeutic targets against the most important protozoan agents of medical interest. PMID- 18422073 TI - [The Instituto de Investigaciones Medicas Alfredo Lanari celebrates its 50 years of existence]. PMID- 18422074 TI - [Alphonse Laveran and malaria]. PMID- 18422075 TI - [Basic investigation according to Hans Selye]. PMID- 18422076 TI - [Seroconversion of hepatitis C in blood donors]. PMID- 18422077 TI - [An almost fatal asthma crisis induced by glucocorticoids. Sometimes it happens]. PMID- 18422078 TI - [Updates in pediatric neurology. Introduction]. PMID- 18422079 TI - [Rett syndrome: 50 years' history of a still not well known condition]. AB - Since it was first described by Andrea Rett 50 years ago, Rett syndrome (RS) has been the subject of further investigations, nonetheless it continues to be a not well known condition. Our own experience and an updated literature review on RS is presented. RS is a severe dominant X chromosome-linked neurodevelopmental disorder with a characteristic clinical picture that mostly occurs in girls, most of the cases are sporadic and genetically determined. The diagnosis of RS is made based on observation and clinical assessment. Main clinical features are mental retardation, behavioural changes, stereotypes, loss of speech and hand skills, gait apraxia, irregular breathing with hyperventilation while awake, and frequent seizures. The internationally established criteria are reviewed. RS is caused by mutations in MECP2 in the majority of cases, but a proportion of atypical cases may result from mutations in CDKL5, particularly the early onset seizure variant. However, the molecular pathogenesis of this disorder remains unclear, as well as the relation between the mutations in MECP2 and other neurodevelopmental disorders. Neuroimaging, neuropathological and biochemical findings in RS are reviewed. Besides symptomatic treatment, no therapeutic trials have shown effectiveness. Some perspectives in the treatment of RS have been provided by a recent work showing a phenotypic reversal by activation of MeCP2 expression in a mouse model. PMID- 18422080 TI - [Neuroprotection in perinatal hypoxic-ischemic encephalopathy. Effective treatment and future perspectives]. AB - The aim of this paper is to review the results of recent clinical studies of some therapies that have demonstrated a neuroprotective effect in perinatal hypoxic ischemic encephalopathy (HIE) and to present the future perspectives of other clinical and basic research investigations. THERAPIES WITH DEMONSTRATED CLINICAL EFFICACY: ALLOPURINOL: It blocks the production of free radicals following hypoxia-ischemia. In a recent study, infants with hypoplastic left heart syndrome treated with allopurinol, but not those with other congenital cardiopathies, had significantly less number of complications than controls, including death, seizures, coma or cardiac events. OPIOIDS: In another recent study, newborns with HIE treated with morphine or phentanyl, had less severe brain damage on MRI and a better neurological outcome. HYPOTHERMIA: Both local (head cooling) or systemic (whole body) hypothermia have a neuroprotective effect in selected newborns with HIE. FUTURE PERSPECTIVES: ANTIEPILEPTIC DRUGS: They have multiple mechanisms of action that can block the biochemical cascade of neuronal damage in HIE. OTHER THERAPEUTIC MODALITIES: Among them the following should be emphasized: combined neuroprotective treatments, growth factors, genetic therapies, stem cell transplant, and neuroprotective immunization. In conclusion, a better knowledge of the molecular mechanisms of HIE pathogenesis and better clinical studies of neuroprotective therapies will open new possibilities aplicable to clinical practice. These advances will undoubtedly improve the prognosis of newborns with HIE. PMID- 18422081 TI - [Sleep disorders in the newborn]. AB - Sleep disorders are classified in dyssomnias, parasomnias, sleep disorder associated with medical and psychiatric disorders and proposed sleep disorders. Only the parasomnias have been studied as such in the newborn period. The parasomnias that occur in this age group are infant sleep apnea, congenital central hypoventilation syndrome, sudden infant death syndrome, and benign neonatal sleep myoclonus. Infant sleep apnea includes three entities: (1) apnea of prematurity, (2), apparent life threatening episodes with apnea and (3) obstructive sleep apnea. Congenital central hypoventilation syndrome can be associated with other autonomic system illness, such as Hirschsprung disease (Haddad syndrome) and neuroblastoma. The implementation of the supine sleep position and smoking free homes has diminished the frequency of sudden infant death syndrome. Benign neonatal sleep myoclonus should be considered in all newborns with a normal exam between the episodes when they always occur during sleep. This entity may be mistaken for status epilepticus, because it is associated with increases in heart rate. Benzodiazepines prolongs the duration of the episodes. PMID- 18422082 TI - [Inborn errors of metabolism with neurological manifestations in the neonatal period]. AB - Congenital metabolic diseases are considered as rare diseases because of their low incidence and their clinical symptoms at onset. Sometimes they can just begin in the neonatal period. Their progressive knowledge and the availability of specific and sensitive biochemical procedures allow us to diagnose many congenital metabolic diseases, which were not recognized some years ago. We reviewed the 52 patients with congenital metabolic diseases diagnosed during the last 25 years in our centre, evaluating the clinical presentation, neurological symptoms, complementary exams and clinical evolution. The mean age at onset of symptoms was 5 days and the mean age at diagnosis was 88 days of age. We considered a first group of 36 patients with inborn errors of intermediary metabolism, in whom hypotonia, weight loss and seizures are the main symptoms. The second group was composed of 8 patients with defective energy metabolism, who showed abnormal respiratory rhythm and hypotonia. Finally, we considered 8 patients with diseases of the complex molecules, who presented with hypotonia and cataracts as common symptoms at onset. The more common neurological symptoms in this period were hypotonia (60%), sensorial deficit (35%) and refractory seizures (23%). The complementary laboratory tests in the first phases of the diseases allowed us to suspect a congenital metabolic disease especially among intermediary and energy defects. EEG and CSF samples were important to diagnose some inborn errors of intermediary metabolism. In the first steps, the neuroimaging was less orientative, even if it allow the exclusion of other diseases. More than half of the patients with inborn errors of metabolism with onset in the neonatal period died within the first two years of life. It is really important to suspect these diseases in the neonatal period so as to achieve an early diagnosis and prompt therapy, specially with cofactors, that can reduce the morbimortality. PMID- 18422083 TI - [Pervasive developmental disorders. Clinical and genetics aspects]. AB - Pervasive developmental disorders (PDD) encompass a heterogeneous group of children with deficits of verbal and non-verbal language, social communication, and with a restricted repertoire of activities or repetitive behaviours. The frequency in general population is considered 27.5/10,000. In this study, we analyzed the clinical and genetic aspects of Autism, Asperger Syndrome, PDD Not Otherwise Specified, Rett Syndrome and Childhood Disintegrative Disorder. We analyzed clinical, behavioural and neuropsychological features. We revised different medical genetics associated conditions and divided the genetics aspects of pervasive developmental disorders into two groups: Syndromic forms (around 20%) and non syndromic forms (currently proposed to be 80%). The early recognition of pervasive developmental disorders and the diagnosis of specific associated syndromes allow early therapy, correct genetic counselling, and follow up anticipating possible complications related to the entity. Finally, although the genetic bases of autism have not yet been identified, the following candidate genes have been proposed: 15q, 2q, 17q, 7q, 12q, and X related genes, among others; which are analyzed in this study and will allow a better understanding of these disorders in the future. PMID- 18422084 TI - [Cerebral palsy]. AB - The term cerebral palsy (CP), is used for a great number of clinical neurological syndromes. The syndromes are characterized by having a common cause, motor defects. It is important, because they can cause a brain damage by presenting motor defects and some associated deficiencies, such as mental deficiency, epilepsy, language and visual defects and pseudobulbar paralysis, with the non evolving fact. Some authors prefer using terms such as "non-evolving encephalopathies". In the treatment the utility of prevention programs of early stimulation and special rehabilitation methods, and treatment of associated deficiencies such as epilepsy, mental deficiency, language, audition and visual problems, and the attention deficit improve the prognosis in an important way. The prognosis depends on the severity of the disease and the associated manifestations. PMID- 18422085 TI - [Syndrome of cerebral non-dominant hemisphere]. AB - In this review the meaning of cerebral hemispheric function is discussed with special emphasis in non-dominant cerebral hemisphere and particularly in the lesion of commissural, association, projection of white matter fibers. Clinical characteristics depend on etiology, localization, extension and particularly the period of brain development. The following are common findings in these patients: left hemiplegia, abnormal prosody and non verbal communication, anomalies visiospatial perception, organization, and social interaction. Similar symptoms are also present in some developmental disorders such as attention deficit hyperactive disorders and Asperger syndrome. We discuss seven cases of isquemic cerebrovascular accidents with the objective of demonstrating the more frequent and persistent findings in the acquired lesion. All patients had complete neurological and neuropsychological evaluation, and were followed up for more than two years. We conclude that motor sequel is less severe than cognitive and affective consequences. PMID- 18422086 TI - [Genetics of epilepsy]. AB - In the last few years, the presence of specific genetic abnormalities leading to some of the classical epileptic syndromes has been clearly elucidated. This means that for the first time, it has become possible to create a strong relationship between the presence of specific genomic and/or proteomic abnormalities and epileptic syndromes previously considered to be "idiopathic". The majority of such genetic defects have been found in genes coding for either ion channels and/or membrane receptors, a fact that somehow seems to confirm the previously postulated importance of the latter structures in the electrochemical activity of neurons. This review will focus on the genetic and clinical aspects of such conditions. Some of the most relevant data suggesting the existence of additional genetic defects in many other epileptic syndromes will also be briefly reviewed, even though a definitive relationship to many of them has not yet been established in the form of specific gene defects. In addition, the worrisome fact that despite the importance of such advances, their application in routine clinical practice remains very limited will be emphasized, in particular in the pharmacological management of most patients. Finally, a brief discussion about the intriguing possibilities of such findings, including the development of neuro pharmacogenomics plus several ethical issues, will also be attempted. PMID- 18422087 TI - [Surgical indications in pediatric epilepsy]. AB - With the goal of presenting follow-up data from patients seen at the Refractory Epilepsy out-patient clinic of the Hospital de Clinicas de Porto Alegre (HCPA), Brazil, we reviewed the literature on childhood and adolescent epilepsy and epileptic syndromes with indication for surgical treatment. The International League Against Epilepsy's Subcommission for Pediatric Epilepsy Surgery recommends surgical evaluation for cortical dysplasia, tuberous sclerosis complex, polymicrogyria, hypothalamic hamartoma, hemispheric syndromes, Sturge-Weber, Rasmussen, Landau-Kleffner and other situations such as tumors or cerebrovascular injury. The Subcommission proposes the creation of referral centers with functional methods of evaluation, structural imaging, and a multidisciplinary team that includes neurologists, neurosurgeons, neurophysiologists, neuropediatricians, neuropsychologists, pathologists, neuroradiologists and eletroneurophysiology technicians. Creating these centers in developing countries is possible with the cooperation of other regions. Studies have shown improvement in quality of life with less frequent seizures. At HCPA, we provide evaluations and ongoing clinical management of patients with medically refractory epilepsy. Epilepsy surgery is potentially curative and is particularly useful for the pediatric population. PMID- 18422088 TI - [Migraine and other primary headaches in children and adolescents]. AB - Migraine diagnosis only relies on clinical characteristics of the episodes and therefore on the doctor's skill and experience. It is recognized that migraine inclusively in the pediatric group is underdiagnosed and inadequately treated. The International Headache Society recently reviewed the international headache classification and incorporated some clinical criteria according to the different age groups. Pediatricians and pediatric neurologists now have a new document and should become familiar with it. This paper discusses these new criteria for migraine and other primary headaches. PMID- 18422089 TI - [Vertigo in children]. AB - Vertigo is uncommon in children, but it is usually underecognized and it has not been well referenced in the medical literature until the 1980s. It is defined as a subjective sensation of movement of the body or the surrounding, and it is a symptom that may have multiple causes. This paper reviews the different types of vertigo in children, emphasizing their clinical features that may be useful in establishing the diagnosis. Of particular relevance is an accurate clinical history and a complete otologic and neurologic examination. More common causes of pediatric vertigo are otitis media and migraine variants, particularly benign paroxysmal vertigo. An appropriate approach to the child with vertigo begins with the clinical suspicion, based on a detailed clinical history and examination, and specific tests. Diagnosis should be followed by adequate symptomatic treatment and management of the underlying specific causes. PMID- 18422090 TI - [Syncope in infants and adolescents]. AB - AIM: To review the syncope in infants and adolescents as well as its etiology, clinical manifestations, physiopathology, diagnostic methods and treatment. DEVELOPMENT: The syncope is a clinical entity characterized by sudden and transient loss of consciousness and postural tone, with a quick and complete recovery. The syncope is frequent in children and adolescents. The most common cause is neurocardiogenic (vasovagal), which has a benign evolution. Other causes may be neurological, cardiac and metabolic diseases, with a variable prognosis, which may come to be severe and potentially lethal. Evaluation of the syncope is mainly based on present clinical history, and on a detailed clinical examination. Basic complementary exams may help on the diagnosis, as a glycemia test, determination of electrolytes (sodium, potassium, magnesium), a hemogram and an electrocardiogram. The Tilt Test is "the" test to confirm the neurocardiogenic syncope. Treatment is indicated in syncopes with recurrent episodes or with risk of physical lesions. Prevention and education are indicated for all patients. Specific treatment should be individually given to each patient. In the case of neurocardiogenic syncope, beta-blockers, alpha-adrenergic stimulants, serotonin reuptake inhibitors, and fludrocortisone are used. In the frequently recurrent neurocardiogenic syncope, when preventive actions are not enough or there is risk of lesions in consequence of falls, beta-blockers, alpha-adrenergic stimulants, serotonin reuptake inhibitors, and fludrocortisone are used, as well as a pacemaker for severe cases non-responsive to pharmacological treatment. CONCLUSION: The syncope is a frequent clinical entity in infancy and adolescence, with benign evolution but potentially lethal. Diagnosis and correct etiology are important for the implementation of specific and early treatment. PMID- 18422091 TI - [Medical and surgical indications of evoked potentials]. AB - Evoked potentials (EP) are more sensitive than specific, safe, objective and reproducible neurophysiologic techniques to assess integrity of peripheral and central sensory and motor pathways. The integrity of pathways are determined by measurement the latency of the recorded and digitally averaged electrical events elicited after adequate stimulation. EP are medically and surgically recommended in conditions in which the diagnoses, determination of neurological outcome in comatose and under anesthesia patients, and determination of the course and evaluation of treatment are uncertain with neurologic examination and other neurodiagnostic studies. PMID- 18422092 TI - [Functional magnetic resonance]. AB - Functional magnetic resonance imaging (fmri) is a non-invasive functional brain mapping technique ased on blood oxygenated level dependent signal (BOLD). It is obtained during T2* weighted imaging MRI studies. fmri aids in the localization of language and motor function for patients candidates for epilepsy surgery, and has up to a 90% concordance with WADA test. Even though synchronous recording of fmri and EEG is technically challenging, it shows a discordant pattern of activation when compared with EEG or electrocorticography. This finding could be due to complex epileptic networks, and overall this study technique has the potential to contribute to further research into epileptic network and epilepsy physiopathology. PMID- 18422093 TI - [Neuroendoscopy. Its usefulness in the hydrocephalus management of children in developing countries]. AB - Hydrocephalus is a huge burden, specially in the poor countries with a big proportion of sick children with this condition, against the countries with good sanitarian conditions. In Latinamerica the most common etiological factors of hydrocephalus are associated with bad prenatal controls, neonatal sepsis and consequent meningitis and ventriculitis. The hydrocephalus is a consequence of the damage in the circulation of the cerebrospinal fluid (CSF). Neuroendoscopy specially third ventriculostomy, associated in some patients with choroid plexus cauterization, represents an alternative method to manage this condition avoiding shunts and its complications. Endoscopic third ventriculostomy (ETV) alone or with choroid plexus cauterization (CPC) is an emerging and very useful technique to manage hydrocephalus specially in children with: ventriculitis as a consequence of meningitis, myelomeningocele, obstructive hydrocephalus idiopathic or seconday to posterior fossa tumors. One hundred consecutive children (100) underwent ventriculoscopy preceding EVT as an initial treatment of hydrocephalus since January 2001 to July 2006, median age was 15.5 months with a range between 2-48 months, 55% are males of our Institution in Valencia, Venezuela. We describe the normal and pathological ventricular anatomy with emphasis in the endoscopic technique and its challenges. We present how the EVT works to prevent the shunt implantation. In the future, neuroendoscopy prevents the use of shunts to treat hydrocephalus in children, avoiding its complications and cost. Neuroendoscopy is a neurosurgical technique everyday most common to manage hydrocephalus and intraventricular lesions in the world. We report an algorythm to use in children younger than 1 year of age with postinfectious hydrocephalus (PIHC) or associated with myelomeningocele. We stress the use of EVT as an alternative because of its low cost to treat hydrocephalus avoiding the complications and dependency of shunts. PMID- 18422094 TI - [The TRITON-TIMI 38 trial]. PMID- 18422095 TI - [Percutaneous heart valves: clinical role]. AB - Percutaneous valve therapy is said to be the last frontier in interventional cardiology. Over the past few years the work of decades has come to fruition. The first percutaneous implantation of a cardiac stent-valve in a human was reported by Bonhoeffer and colleagues in 2000. The first percutaneous aortic valve placement was performed by Cribier in 2002 using an antegrade approach in a patient with critical aortic stenosis. Attempts at percutaneous mitral valve repair are as varied as the pathophysiological mechanisms of mitral regurgitation. We are now embarking on a new era in the treatment of valvular heart disease with the introduction of percutaneous and minimally invasive devices and techniques to address valve dysfunction without conventional surgical repair/replacement. Pathology of all four cardiac valves has now been treated in early stage clinical feasibility (pilot) trials. There are at least 30 percutaneous valve programs currently being developed by 24 different companies. Two of the percutaneous aortic valve devices and five of the mitral valve devices are in various stages of clinical trials, ranging from feasibility to pivotal at the present time. Large-scale trials comparing percutaneous valve replacement and repair with surgery are just beginning. In this article we will look at the various technologies and advances in percutaneous valve replacement and repair, with an emphasis on those devices that are currently being utilized in the clinical realm. PMID- 18422096 TI - [Cardiac engineered valves: state of the art]. AB - Engineered heart valves promise to be the ideal heart valve replacement: they have the potential to grow and repair within the host, to minimize inflammatory and immunological responses and to limit thromboembolism. In particular, pediatric patients with congenital defects would greatly benefit from growing replacement materials by reducing the need for surgical reinterventions due to the lack of prosthetic material growth. In fact, viable cells included in engineered heart valves can theoretically adapt to a growing and changing environment exactly as a native biological structure. Engineered heart valves represent thus a novel experimental approach to create autologous, living replacements with the potential to grow, regenerate, and remodel. The aim of this paper is to present a review concerning the research efforts to create a viable engineered heart valve. PMID- 18422097 TI - [A database in the coronary care unit to assess quality of care of acute myocardial infarction]. AB - BACKGROUND: In the context of an operative network for acute coronary syndrome consisting in telemedicine equipment and 24-h catheter facility availability, a database was worked out to verify clinical pathways, timeline, prevalence of cardiovascular risk factors, diagnostic and therapeutic choices, and clinical results. The aim of this study was to evaluate the adherence to guidelines in the real world in order to assess quality of care. METHODS: We implemented a database in the preexisting hospital intranet due to open source interactive pages. From international guidelines we obtained performance measures for each step of the care process, in order to have a continuous and updated evaluation of our healthcare performance. After an accurate analysis of the results of the first 2 year data collection, we modified the performance measurement set to optimize our database. All data were analyzed with statistical program SPSS. RESULTS: A total of 1066 patient characteristics within a period of 5 semesters were analyzed: trends for each of the 11 performance measures were evaluated. In 852 patients clinical and angiographic characteristics were analyzed: cardiovascular risk factors and TIMI risk score for ST-elevation myocardial infarction patients, different pathways and first-aid timeline. Data about pharmacological therapy on admission, during intensive care unit stay and at discharge were also collected. CONCLUSIONS: The present study assesses the utility of continuous monitoring and of verifying quality of care with a dedicated database in order to evaluate adherence to guidelines and to promote optimization of clinical practice. PMID- 18422098 TI - [Permanent registries: just an unmet dream for the Italian cardiologist?]. PMID- 18422099 TI - [Myocardial revascularization and radial artery: different surgical strategies and short-term results]. AB - BACKGROUND: The radial artery is routinely used as a graft for surgical arterial myocardial revascularization. The proximal radial artery anastomosis site remains unknown. In this study, we analyzed the short-term results and the operative risk determinants after having used four different common techniques for radial artery implantation. METHODS: From January 2000 to December 2004, 571 patients underwent coronary artery bypass grafting with radial arteries. Data were analyzed for the entire population and for subgroups following the proximal radial artery anastomosis site: 140 T-graft with the mammary artery (group A), 316 free-grafts with the proximal anastomosis to the ascending aorta (group B), 55 mammary arteries in situ elongated with the radial artery (group C) and 60 radial arteries elongated with a piece of mammary artery and anastomosed to the ascending aorta (group D). RESULTS: The mean age was 53.8 +/- 7.7 years; 55.5% of patients had a previous myocardial infarction and 73% presented with a satisfactory left ventricular function. A complete arterial myocardial revascularization was achieved in 532 cases (93.2%) and 90.2% of the procedures were performed under cardiopulmonary bypass and cardioplegic arrest. The operative mortality rate was 0.9%, a postoperative myocardial infarction was diagnosed in 19 patients (3.3%), an intra-aortic balloon pump was used in 10 patients (1.7%) and a mechanical circulatory device was implanted in 2 patients. The radial artery harvesting site remained always free from complications. The proximal radial artery anastomosis site was not a determinant of early hospital mortality. Group C showed a higher risk of postoperative myocardial infarction (p = 0.09), together with female gender (p = 0.003), hypertension (p = 0.059) and a longer cardiopulmonary bypass time. CONCLUSIONS: The radial artery and the mammary artery can guarantee multiple arterial revascularization also for patients with contraindications to double mammary artery use. The four most common techniques for proximal radial artery anastomosis are not related to a higher operative risk and they can be used alternatively to reach the best surgical results. PMID- 18422100 TI - [Appropriateness of simultaneous carotid endarterectomy and coronary revascularization]. AB - BACKGROUND: The presence of significant carotid artery disease in patients undergoing coronary artery bypass grafting has been reported to be as high as 17%. The optimal management of patients with significant coronary and carotid artery disease remains controversial. In this study, we analyze our recent experience with patients who underwent synchronous carotid endarterectomy (CEA) and coronary artery bypass grafting. METHODS: We reviewed the early outcome of 68 patients (56 males, 12 females, mean age 71.1 years, range 53-88 years) who underwent simultaneous CEA and coronary artery revascularization between January 2005 and June 2007. The frequency of unstable or ulcerated plaques was determined in symptomatic and asymptomatic patients. RESULTS: Death for myocardial infarction occurred in 3 patients (4.4%). Stroke was found in 1 patient (1.4%). Combined 30-day stroke/mortality rate was 5.8%. The frequency of unstable or ulcerated plaques was 60.3% (41/68). An unstable stenosis was present in 23 out of 42 asymptomatic patients (54.7%). CONCLUSIONS: Patients suffering from a concomitant coronary and carotid artery occlusive disease represent a high-risk population whose management is still controversial. A modern approach to combined CEA and coronary artery bypass grafting may be safe. The high frequency of unstable carotid lesions in asymptomatic patients suggests to treat every stenosis > 75% in candidates to coronary artery bypass grafting. Carotid artery stenting should be avoided in the majority of cases, considering the possibility of unstable carotid stenosis and the atherosclerotic involvement of aortic arch. PMID- 18422101 TI - [Pitfalls of cardiologic consultation in internal medicine: an obvious case of alcoholic liver cirrhosis]. PMID- 18422102 TI - [A complicated case of purulent pericarditis secondary to pneumonia]. AB - Purulent pericarditis in the antibiotic era has become an uncommon entity. It is typically an acute illness with a high mortality (about 30%) related to cardiac tamponade, a complication that accounts for 42-77% of cases. Few cases of purulent pericarditis associated with tamponade have been published in the literature. In this paper we describe a complicated case of purulent pericarditis secondary to left pneumonia. In particular, we emphasize the importance of early recognition, prompt institution of appropriate antibiotic therapy and early drainage for survival. PMID- 18422105 TI - [Implications for clinical practice of the new definition of myocardial infarction]. PMID- 18422103 TI - [Pseudoaneurysm late after surgical repair of aortic coarctation]. PMID- 18422106 TI - [Analyses of spectral emissivity in radiation temperature measurement]. AB - The complexity of the spectral emissivity of actual surfaces is the key point in the research and applications of radiation temperature measurement, resulting in the difficulty in the achievement of the temperature measurement. In the present paper, based on the discussions of the Taylor expansion, the non-dimension wavelength and the exponent, the authors describe the mathematical expression of the spectral emissivity of actual surfaces, and establish the general spectral emissivity function. Through the fitting of experimental data of the spectral emissivities of different metals at different temperatures, the applicability of the spectral emissivity function is verified which especially becomes the fundamental in the research of primary spectrum pyrometry. PMID- 18422107 TI - [Temperature measurement of DC argon plasma jet]. AB - The electron temperature of DC arc plasma jet is an important parameter, which determines the characteristics of plasma jet. The measurement of emission spectrum was performed to obtain the spectral intensities of some Ar lines and the method of diagrammatic view of Boltzmann was adopted to calculate the electron temperature. The results indicated that the electron temperature dropped at different speed along with the axes of the plasma jet and rose rapidly when the current was increased, and it also rose when the flowrate of argon was increased. PMID- 18422108 TI - [Absorption and scattering characteristics of human benign prostatic hyperplasia tissue with Ti: sapphire laser irradiation in vitro]. AB - The optical properties and their differences of human benign prostatic hyperplasia (BPH) tissues removed using transurethral plasma kinetic resection of the prostate (PKRP) and transurethral vaporization of the prostate (TUVP) at 640, 660, 680, 700, 720, 740, 760, 780, 800, 820, 840, 860 and 880 nm of Ti: Sapphire laser were studied in vitro. The measurements were performed using a double integrating-sphere setup, and the absorption and scattering properties were assessed using the inverse adding-doubling method. The results of measurement showed that the absorption coefficients and reduced scattering coefficients of BPH tissues removed using PKRP and TURP obviously decreased with the increase in the wavelength for thirteen different laser wavelengths. The absorption coefficient and reduced scattering coefficient of BPH tissues removed using PKRP at a certain laser wavelength were obviously smaller than that of BPH tissues removed using TUVP at the same laser wavelength. The maximum absorption coefficient and maximum reduced scattering coefficient of BPH tissues removed using PKRP and TURP were respectively (0. 885 +/- 0. 022) and (0.955 +/- 0.024)mm(-1), and (1.564 +/- 0.039) and (1.658 +/- 0.042)mm(-1) at 640 nm, their differences were respectively 7.91% and 6.01%, and the minimum absorption coefficient and minimum reduced scattering coefficient of BPH tissues removed using PKRP and TURP were respectively (0.443 +/- 0.011) and (0.455 +/- 0.011) mm( 1), and (1.117 +/- 0.028) and (1.197 +/- 0.030)mm(-1) at 640 nm, their differences were respectively 2.71% and 9.13%. The maximum difference in the absorption coefficients of BPH tissues removed using PKRP and TURP is 8.95% at 660 nm, and the minimum difference is 1.75% at 860 nm. The maximum difference in the reduced scattering coefficients of BPH tissues removed using PKRP and TURP is 9.13% at 800 nm, and the minimum difference is 6.01% at 640 nm. PMID- 18422109 TI - [Measurements of density distribution of high energy electrons by emission spectroscopy in pulsed discharge reactor]. AB - Pulsed streamer discharge can generates high energy electrons with energy higher than 11. 03 eV and enough to excite N2 to its C3 pi(u) state from the ground state, so the density of the high energy electrons can be obtained by diagnosing the relative emission spectrum intensity of the N2 (C 3 pi(u) --> B 3 pi(g)) emitted from the pulsed streamer discharge. In the present paper, the density distributions of high energy electrons in the wire-plate DeSO2 reactor with pulsed streamer discharge were studied in air at room temperature and 1 atm, and the influences of pulsed voltage and wire-to-wire spacing on the density distributions of high energy electrons were also studied. The experimental results show that high-energy electrons mainly centralize around the discharge wire of the wire-plate reactor, and the electron density reduces with the increase of the distance from the discharge wire. The pulse voltage has a great influence on the high energy electron density, and the high energy electron density increases linearly with increasing the pulsed voltage. Under a fixed wire to-plate spacing, when the wire-to-wire spacing is 0. 6-1 times the wire-to-plate spacing, the density distributions of the high energy electrons are more uniformity. PMID- 18422110 TI - [Spectral study of the effects of glasses lenses' color depth on stereoscopic vision]. AB - Glasses with zero focal-length lens and different shades of red, yellow and blue were studied in terms of absorbance by UV-Vis spectral absorption method. The effect of lens with different shades of the same color on stereoscopic vision was comparatively studied from the medical view. The study has shown that, for yellow and blue lens, there is no difference in the absorbance between the deep-color and light-color lens in UV; at 550 nm, the absorbance of deep-color lens is higher than that of light-color lens. Whereas, for red lens, the absorbance of light-color lens in UV is higher, and at 550 nm, still higher than that of deep color. Those with light-color glasses of yellow or blue have much better stereoscopic vision than those with deep-color glasses, while the case is inversed as to those wearing red glasses. The conclusion can be drawn that the effect of lens with different shades on stereoscopic vision depends on the absorbance of lens at 550 nm, and the higher the absorbance, the stronger the effect. PMID- 18422111 TI - [Influence of pressure on plasma temperature in air/argon dielecteic barrier discharge]. AB - Electron excitation temperature and molecule vibrational temperature in argon/air dielectric barrier discharge (DBD) at different gas pressure with water electrodes were studied by using optical emission spectra. The spectral lines of Ar I 763. 51 nm(2P6 --> 1S5) and Ar I 772.42 nm(2P2 --> 1S3) were chosen to calculate electron excitation temperature by the relative intensity ratio method. The emission spectra of nitrogen band of second positive system ( C3 pi(u) --> B3 pi(g)) were measured at the same time. The molecule vibration temperature was estimated by the emission intensities of different bands with delta(nu) = -1, delta(nu) = -2, and delta(nU) = -3 in nitrogen band of second positive system, using Boltzmann's plot method. In addition, the relative line intensities of nitrogen (0-0) band of first negative system at 391.4 nm and (0-0) band of second positive system at 337.1 nm were also measured to study the variation of electron energy. It was found that the electron excitation temperature decreased from 4 700 to 3 300 K and the molecule vibrational temperature decreased from 3 200 to 2 900 K with increasing gas pressure from 20 to 60 kPa. Besides, the ratio of I(N2+)/I(N2) also decreased with pressure increasing from 20 to 60 kPa, indicating that the average electron energy decreases with the gas pressure increasing. These results are of great importance to the study of plasma dynamics of dielectric barrier discharge and also to the underlying industrial applications. PMID- 18422112 TI - [Synthesis and spectroscopy of composite materials made from lanthanide substituted polyoxotungstate and hemicyanine derivative]. AB - The organic composite materials formed by Keggin lanthanide-substituted polyoxotungstate anion [Ln(SiW11 O39)2 ]13- (in which Ln = La, Ce, Pr, Nd, Sm) and (E)-N-methyl-4-(2-(4'-dimethylaminophenyl)ethenyl)pyridinium cation with the stoichiometry of (C16 H19 N2 )10 K3 [Ln(SiW11 O39)2], have been synthesized and characterized by elemental analyses, thermogravimetric and differential thermal analyses, and IR, UV-Vis, and fluorescence spectroscopy. The results of spectra show that the title compounds have Keggin structure, and inorganic-organic composite materials are formed between the hemicyanine cations and the heteropoly anions, and the charge transfer occurs from the hemicyanine cations to the the heteropoly anions in title complexes. The compounds reported in this paper are interesting as optical materils. PMID- 18422113 TI - [Spectral characteristics of biological soil crusts in Gurbantonggut Desert, Xinjiang]. AB - Large areas of desert in arid and semi-arid regions are covered by biological soil crusts. Tracking the presence, distribution and disappearance of biological soil crusts is important for ecosystem management of desert regions and provides highly valuable information about desertification and climate change studies in arid and semiarid environments. The objective of the present paper is to analyze the unique reflectance spectral features of three types of biological soil crusts (moss, lichen and algae) relative to bare sand and dry plant. The measurements for the reflectance of bare sand, dry plant and biological soil crusts derived from the field survey were performed in the laboratory under dry condition. The reflectance of biological soil crusts after they were hydrated with 0.5 and 1.0 mm water respectively was also measured in the laboratory. It was found that the reflectance of crusts was much lower than that of bare sand throughout the Vis SWIR region. Moreover, the reflectance of crusts exhibited a slightly flattening plateau between 600 and 750 nm, attributed to the absorption of photosynthetic pigments. Based on the spectral features of biological soil crusts, it was proposed that continuum removal technique could be used to retrieve the coverage of biological soil crusts. A significant correlation between the minus logarithm of the absorption depth of the continuum removal spectra and the coverage of biological soil crusts was found (r2 = 0.990 7). The liquid-water hydration experiment demonstrated that the absorption of photosynthetic pigments was strengthened after hydration, making the detection of biological soil crusts much easier. PMID- 18422114 TI - [Visible light reflectance spectrum for measurement of cancerous tissue]. AB - A goal the authors always pursue is to realize diagnosis of precancer in vivo, real-time and non-invasive. In the present paper, results of diagnosis of certain cancer were obtained by pathological analysis of several samples, and then the visible light reflectance spectrum property of cancerous tissue was acquired by the comparison with the reflectance spectrum of normal tissue in the same organ. It was found that the main result of change in the reflectance spectrum was caused by the increases in hemoglobin and deoxygenated hemoglobin. The cancerous tissue has a lower reflectance in visible spectrum and has the strongest change in the absorption at 630 nm. Absorption peaks in the reflectance spectrum indicated that there are abundant of oxygenated hemoglobin and deoxygenated hemoglobin in the cancerous tissue. It is the same characteristics as in the cancerous tissue. It was fully indicated experimentally that visible light reflectance spectrum can distinguish normal and cancerous tissue. PMID- 18422115 TI - [Application of wavelet transform to the infrared diffusion reflectance spectrum of rocks]. AB - The infrared diffuse reflectance spectra of hydrocarbon source rocks with different particle sizes were measured. The result indicated that the absorbency of the raw spectrum decreased with the reduction of particle size, but the relationship turned to be reverse after we pretreated the original spectra by using wavelet transform to eliminate the background and calibrate the baseline drift, both of which were caused by scattering. The reversed relations showed that the spectral lines were influenced deeply by the scattering of the samples. So the particle size of the samples to be measured and the particle size of the model samples must be consistent to reduce the error. The low frequency part of the spectrum filtrated by wavelet transform corresponds to the scattering, and the authors used it to set up a model at the latent absorbance wave number (near 2 820 cm(-1)) to forecast the particle size. By comparing this model with the other model based on the original spectrum the authors found that the pertinence of the anterior model is higher than the latter one and the value reaches 0.999 7. So the authors can accurately forecast and control the distribution of the particle size by this model, which can be used to improve the accuracy in the quantitative analysis of the infrared reflectance spectrum. Also the study validated that both scattering and absorption coefficients are inversely correlated with the particle size. PMID- 18422116 TI - [Broad excitation band alkaline-earth silicate luminescent materials activated by rare earth and its applications]. AB - Series of novel broad excitation band phosphors M2 MgSis O7 : Eu, Dy(M = Ca, Sr) were prepared by a high temperature solid-state reaction method. The crystal structure of compound was characterized. And the effects of part substitution of alkaline-earth on crystal structure, photoluminescence spectra and luminescence properties were also investigated. It is found that the excitation band of silicate luminescent materials extend to visible region and they exhibit yellow, green and blue long after-glow luminescence after excited by ultraviolet or visible light. Ca MgSi O7 : Eu, Dy luminescent materials can be excited effectively under the 450-480 nm range and exhibit a strong emission at 536 nm, nicely combining with blue light emitted by InGaN chips to produce white light. This promises the silicate luminescent materials a potential yellow phosphor for white LED. PMID- 18422117 TI - [Ethanol electrooxidation on carbon supported PtSn catalyst: in situ TRFTIR study]. AB - Carbon supported PtSn catalyst (PtSn/C) was prepared by a modified polyol method and characterized by means of XRD. It was showed that the metal particle size was 2.2 nm and the unit cell parameter increased compared with Pt/C. In situ time resolved Fourier transform infrared spectroscopy (TRFTIRS) was used to study the electrooxidation of ethanol on PtSn/C catalyst. COL was the main poison species adsorbed on the active sites to inhibit the further reaction of ethanol electrooxidation. Acetaldehyde and acetic acid were found to be the products of ethanol electrooxidation as competing reactions with ethanol dissociation when the potential was up to 0.3 V, which reduced the poisoning effect. The selectivity of acetic acid among the products was improved with the increase in the potential and reaction time. CO2, which appeared at 0.4 V, was the final product and yielded from the oxidation of COL. The catalytic mechanism of PtSn/C towards ethanol electrooxidation was analyzed based on the results. PMID- 18422118 TI - [FTIR spectroscopic study on carcinoma cells]. AB - The aim of this research is to establish the cell basis for the carcinoma tissue diagnosis by exploring a method to obtain the FTIR (Fourier transform infrared) spectra of the cultured carcinoma cells with FTIR spectroscopy and investigating the special spectral features of the carcinoma cells compared with the carcinoma tissues. In the present paper, the gastric carcinoma tissues confirmed by histology were measured using a Nicolet Magna750-II FTIR spectrometer and the corresponding FTIR spectra were obtained. The cultured gastric carcinoma cells (SGC7901) were centrifuged to provide a small pellet of cells for FTIR analysis. The cell pellet was then placed on a specially designed salt plate made of BaF2. Then the infrared spectra were recorded by the same equipment. Based on the previously established criteria, a comparative study was subsequently carried out between the spectra of the cultured carcinoma cells (SGC7901) and that of the corresponding gastric tissues. Several infrared spectral features of the carcinoma cells were obtained: the different bands between cells and tissues locate in the range of 3 000-3 600 cm(-1) and 1 640 cm(-1) which are the range of the hydroxy stretching and blending bands of H2O. There are more H2O out of carcinoma cells in carcinoma tissues, so the strong bands of H2O cover the distinctive bands of carcinoma cells in carcinoma tissues. Although the carcinoma tissue is more complicated, which might originate from the intrinsic complexity of the tissue, the results suggest that the spectral features of the carcinoma cells can be well reflected by that of the carcinoma tissue. This study shows that the diagnosis of carcinoma tissue by FTIR method exhibits sufficient cell basis. PMID- 18422119 TI - [Study on the identification of some species of Herba Epimedii with FTIR]. AB - Herba Epimedii, the dried aerial parts of Epimedium brevicornum Maxim., E. sagittatum Maxim., E. pubescens Maxim., E. wushanense T. S. Ying, and E. koreanum Nakai, has been widely used in the field of nourishing the kidney and reinforcing 'Yang' for centuries. The descriptions of external appearance of Herba Epimedii are rather simple, so it is difficult to identify the raw materials of Herba Epimedii only with the traditional methods. In the present paper, the methods of FTIR and the second derivative IR spectra were used for the identification of Herba Epimedii, by which, E. brevicornum Maxim. could be identified from E. koreanum Nakai, and so E. sagittatum (Sieb. et Zucc.) Maxim. from E. acuminatum Franch., as well as E. pubescens Maxim. from E. stellulatum Stearn, E. baojingensis Q. L. Chen et B. M. Yang, E. zhushanense K. F. Wu & S X. Qian and E. myrianthum Stearn. According to the features of the IR spectra of each species, a method based on FTIR for the identification of Herba Epimedii was established, and it conduced to solve the confusion of species of Herba Epimedii. PMID- 18422120 TI - [FTIR study on the hydrocarbon-generation mode of Jurassic coals from Turpan-Hami Basin, Xinjiang]. AB - High temperature simulation experiment by quartz cuvette technique and Fourier transform infrared spectroscopy (FTIR) was used for the study of the chemical structures of kerogen from the Early Jurassic coal in the J(1b) coal bed in Kekeya, Turpan-Hami Basin, Xinjiang at different temperatures. Combining the microscopic statistical results of macerals with the change of IR peaks and corresponding IR parameters with increasing simulation temperature, the hydrocarbon-generation characteristic and mode of the algae coal from Kekeya were estimated. The results indicate that the coals in question are a satisfactory kind of source rock for oil and gas, since their main component is desmocollinite and also contains some algae and many small sporogonium. As the temperature becomes higher and higher, the offshoots of the aromatic structure keep falling off. The heteroatomic compounds and aliphatic structure which have lower activation energy are gradually separated out, inducing that the hydrocarbon generation potential of kerogen decreas. This process begins at 200 degrees C or so, bulges at 350 degrees C, and nearly comes to an end at 500 degrees C. What's more, kerogen tends to be graphitized after 500 degrees C-cores of aromatic structure aggregate and arrange more and more serriedly; this reaction looks quite obvious when it's heated to 600 degrees C. PMID- 18422121 TI - [An FTIR and XPS study of immobilization of chromium with fly ash based geopolymers]. AB - Immobilization of Cr3+ with fly ash geopolymers was investigated by Fourier transform infrared spectroscopy (FTIR) and X-ray photoelectron spectroscopic (XPS) techniques. The chromium sludge, as Cr(OH)3, was prepared with chemical precipitation method. The amounts of aluminum and silicon leached before and after the chromium sludge addition were measured using ICP-AES. The results suggested that the amounts of silicon and aluminum leached were reduced for the fly ash geopolymers after chromium sludge was incorporated. The decrease of silicon leaching was more pronounced than aluminum. FTIR results showed that the intensity of the main peak shifted into lower and the wave number of Si--O--Si and Al--O--Si became higher. The XPS results indicated that the O(1s) bind energy decreased, Si(2p) and Cr(2p) bind energy increased, while Al(2p) bind energy remained unchanged due to Cr3+ addition. It was also confirmed that the chromium is easily incorporated into the fly ash geopolymers paste, and polymerized with silicate units. The immobilization of Cr3+ using fly ash geopolymers is attributed not only to physical encapsulation, but also to chemical reaction. PMID- 18422122 TI - [Identification of Pericarpium Citri Reticulatae from different years using FTIR]. AB - The Pericarpium Citri Reticulatae from different years were analyzed and identified by Fourier Transform Infrared Spectroscopy (FTIR) and method of relative coefficience compare in this paper. The evidence indicated that the main constituent of the alcoholic extract of Pericarpium Citri Reticulatae was hesperidin, the FTIR spectra of the alcoholic extract of Pericarpium Citri Reticulatae ranged among 2 851, 1 716 and 1 516 cm(-1), and the peak had the obvious tendency to increase while the times prolonged, moreover its second derivative spectra also made corresponding changes among 1 734, 1 517 and 1 276 cm(-1). Compared to the alcoholic extract of Pericarpium Citri Reticulatae in 1 year, their relative coefficience decreased gradually when the years increased. According to the fingerprint character of the alcoholic extract of Pericarpium Citri Reticulatae, it meant that accomplished with the time extended, the hesperidin was increasing, meanwhile, some organic acid and ester were increasing too. Above all, it was proved that FTIR was a fast and simple method to identify the Pericarpium Citri Reticulatae in different years. PMID- 18422123 TI - [FTIR and thermal analysis studies of the effects of additives on thermal decomposition of AP/Al]. AB - The mechanisms for AP of the main compositions of composite propellants with additives such as ammonium oxalate (AO), strontium carbonate (SC) and AO/SC were studied by FTIR and DSC. The analysis of FTIR shows that AO leads to a delay in the temperature for the disappearance of absorption peaks of AP. SC reacts with HClO4 from AP decomposition and produces more stabilized Sr(ClO4)2 in the condensed phase. FTIR proves the production of Sr(ClO4)2. The analysis of DSC shows that the temperature for high decomposition exothermic peak of AP with AO is increased, but there is no effect on the temperature for low decomposition exothermic peak of AP. Both the temperature for the low and high decomposition exothermic peaks of AP are increased by the addition of SC. Although the temperature for decomposition exothermic peak of AP is increased with the addition of AO/SC, the experimental result shows that SC and AO didn't produce synergetic effects for the high temperature decomposition of AP at low pressure. Based on the above experimental results, the mechanisms of inhibiting the decomposition of AP for AO and SC are discussed. PMID- 18422124 TI - [Infrared spectroscopic study on storage substance mobilization of crop seeds in germination]. AB - The traditional method to study the storage substance mobilization of seeds during the process of germination is chemical extraction, which is troublesome and time consuming. In the present paper, Fourier transform infrared spectroscopy was used to study the storage substance mobilization of legume and cereal seeds in germination. The spectral results show that legume seeds (soybean, pea and broad bean) exhibit similar remarkable changes in the region from 1 000 to 1 200 cm(-1) in germination, which suggest that the storage carbohydrates in endosperm of legume seeds are mobilized during the process of germination. Notable changes were observed on amide bands of protein and lipid band in the spectra of germinating cereal seeds (rice, wheat and barley), indicating that storage proteins and lipids in cotyledon were utilized during the process of cereal seed germination. The results suggest that vibrational spectroscopy has the advantages of rapidress and convenience in the study of crop seeds and seed physiology. PMID- 18422125 TI - [Relationship between different milk processing and near infrared spectra]. AB - In the present paper, the effects of different milk processing (homogenizing process, pasteurization) on the milk NIR spectra were discussed. It was found that the raw milk and processed milk show significant difference in the 1 890 nm region, which can be used not only to identify the processed milk, but also to offer the basic theory for NIRS in the quality control researches of milk. The absorbance sharply reduced when the liquid milk was treated by a homogenizer, but the absorbance increased after pasteurization. Raw milks absorbance shows a downtrend in the whole region of spectra with increasing pressure. The changes in fat globules structure finally result in absorbance decline. The commercial milk including remade milk was taken for example to discuss the mechanism of detection. The discriminate analysis calibration was developed by SIMCA method and the accuracy of detection is 98.1% for identifying the reconstituted milk in pasteurized milk between 1 800 and 2 200 nm with the pretreatment method of second derivatived and Norris 5.5. PMID- 18422126 TI - [Progress in study of secondary structure of denaturized protein by FTIR]. AB - Protein is one of the most important living matters, and researches relatedto it have interested so many scientific workers. The secondary structure of protein plays a crucial role in maintaining protein's physiological activity. With the development of technology and the improvement of research technique, there are more and more in-depth studies on protein's microstructure. In recent years, many research works in this field have been done with different kinds of physical and chemical methods and Fourier transform infrared spectroscopy(FTIR)holds an unshaken position due to its unique superiority. The present paper is a general review of the related studies all over the world, and the assignments of the major infrared absorption bands of denaturized protein are introduced, and the spectroscopic features and the analytical methods are mainly reviewed. PMID- 18422127 TI - [Research on structure of MC-nylon 6/aramid composites by spectroanalysis method]. AB - The Kevlar fiber, treated with toluene-2,4-diisocyanate and caprolactam, was used to reinforce MC nylon 6. XPS showed the change in chemical components and spectra after fiber was treated. IR showed that the spectra of Kevlar fiber and MC-nylon 6 were incorporated purely when they were mixed, while the treated Kevlar fiber provided the activation in the ring-opening polymerization of caprolactam, and the amide groups of graft chains could form strong hydrogen bonds with matrix. These graft chains and hydrogen bonds would contribute to improving interfacial bonding between MC-nylon 6 and Kevlar fiber. XRD indicated that the induction of Kevlar fiber had no effect on the cryastalline style of MC-nylon 6 obviously, however, the crystals had better seasonal structure. The crystals of MC-nylon 6/Kevlar fiber composites were more perfect than that of MC-nylon 6/Kevlar untreated fiber composites with the same amount of Kevlar fiber. The increase of Kevlar fiber contributed to forming perfect a spherulite when the mass concentration of Kevlar fiber was less than 2%, however, the more the Kevlar fiber, the less the content of a spherulite when the mass concentration of Kevlar fiber was more than 2%. PMID- 18422128 TI - [Novel synthesis and spectral characterization of nelfinavir]. AB - An efficient and environmentally friendly procedure for the one-pot synthesis of (3S,4aS,8aS)-2-((2R,3R)-3-amino-2-hydroxy-4-(phenylthio)butyl)- N-tert -butyl decahydroisoquinoline-3-carboxamide(VII), the intermediate of nelfinavir, was described, and activating ester was applied to getting nelfinavir(IX). Under the catalysis of potassium hydroxide, benzyl (R)-1-((S)-oxiran-yl)-2-(phenylthio) ethyl carbamate was obtained(IV) in methanol. Then IV and (3S,4aS,8aS)- N-tert butyl-decahydroisoquinoline-3-carboxamide(V) were refluxed in methanol until the reaction was finished. Potassium hydroxide(w(KOH) = 40%) in water was added to remove benzyloxycarbonyl group in water bath with a yield of 89.0%. 3-acetoxy-2 methylbenzoic acid-succinimide ester(II) reacted with VII and acetyl group was removed by dense aqueous ammonia, giving nelfinavir(IX). The vibrations of functional groups of thiIIs compound corresponding to the main infrared absorption peaks were discussed. The molecular ion and quasi molecular ion peaks were obtained via MALDI-TOF MS, and 1H NMR and 13CNMR shifts of this compound were assigned by means of DEPT(distortionless enhancement by polarization transfer spectrum), HMBC(1H detected heteronuclearmultiple bond correlation spectrum), HSQC(1H detected heteronuclear single-quantum coherence spectrum) and DQF-COSY(double-quantum filtered-correlated spectroscopy). The results provided useful information for structure characterization and quality control of nelfinavir. PMID- 18422129 TI - [Synthesis and spectral properties of cuboid-shaped CdS particles]. AB - Cuboid-shaped CdS particles were synthesized with sol-gel method by using thiourea as surface modifier and characterized by X-ray diffraction, transmission electron microscopy, infrared absorption spectrum and fluorescence spectrum. The results show that the sulfur atoms in thiourea molecules coordinate with Cd2+ ions on the surface of the nanocrystals. The thiourea-modified CdS particles have a zinc blende crystal structure and good fluorescent characteristics. The cuboid shaped CdS particles are probably assembled by hydrogen bonding between primary CdS nanocrystals surface-capped with thiourea. The result will promote the researches into molecular self-assembly of nanocrystals and help develop novel luminescence material. PMID- 18422130 TI - [Comparative studies on the characteristics of the Fourier-transform infrared spectra between sturgeon and shark chondroitin sulfates]. AB - The characteristics of the Fourier-transform infrared spectra of sturgeon and shark chondroitin sulfates (CHSs) were comparatively studied. The results show that sturgeon CHS exhibits special vibrations at the wavenumbers of 1 376, 1 344, 1 310, 1 157, 883 and 856 cm(-1). Further analysis shows that shark CHS contains 6-sulfated-CHS, while sturgeon CHS contains 4, 6-disulfated CHS, indicating that sturgeon CHS could have higher biological activities in decreasing the toxicity of medicine and in killing cancer cells. In general, the two kinds of CHSs have similar infrared spectra and groups of acylamino, carboxyl, sulfate, and saccharide ring. But the N-H variable-angle vibration of acylamino group of sturgeon CHS occurs at the higher wavenumber, and the vibration intensity of carboxyl group at 1 415 cm(-1) is also stronger than that of shark CHS. Chemical analysis shows that sturgeon CHS has a higher content of glucuronic acid, suggesting that it probably could be a better kind of medical materials for the bone mineralization. PMID- 18422131 TI - [Progress in labeled immunoassay based on SERS]. AB - Labeled immunoassay utilizing surface-enhanced Raman spectroscopy (SERS) is a new style research technology. The discovery and confirmation of surface-enhanced Raman spectroscopy have made Raman spectroscopy a powerful tool in many research fields. SERS has the advantages such as high sensibility and selectivity, also it is suitable for the study of liquid substance. In recent years, it has shown potential application future in biomedical field, and also developed fast in labeled immunoassay. The principle, specialty, problem and recent advances of labeled immunoassay based on SERS were reviewed in terms of detection limits, non specific adsorption and multi-analyte immunoassay. The latest advancement in the improvement of detection limits was summarized, along with the introduction of some work in our lab, as well as expatiating on the effect of non-specific adsorption. Finally, the development trends and application perspectives were discussed. PMID- 18422132 TI - [AgBr colloids prepared by electrolysis and their SERS activity research]. AB - Ivory-white AgBr colloids were prepared by means of electrolysis. Two silver rods 1.0 cm in diameter and 10.0 cm long were respectively used as the negative and positive electrodes, the aqueous solution of hexadecyl trimethyl ammonium bromide was used as the electrolyte, and a 7 V direct current was applied on the silver rods for three hours. The obtained AgBr colloids were characterized by UV-Vis spectroscopy, transmission electron microscopy, and SERS using a 514. 5 nm laser line on Renishaw 2000 Raman spectrometer. These particles are about nanometer size and their shapes are as spherical or elliptic, with a slight degree of particle aggregation. The UV-Vis spectra exhibit a large plasmon resonance band at about 292.5 nm, similar to that reported in the literature. The AgBr colloids were very stable at room temperature for months. In order to test if these AgBr colloids can be used for SERS research, methyl orange, Sudan red and pyridine were used. It was found that AgBr colloids have SERS activity to these three molicules. For methyl orange, the intense Raman peaks are at 1 123, 1 146, 1 392, 1 448 and 1 594 cm(-1); for Sudan red, the intense Raman peaks are at 1 141, 1 179, 1 433 and 1 590 cm(-1); and for pyridine, the intense Raman peaks are at 1 003, 1 034 and 1 121 cm(-1). It is noticeable that SERS of methyl orange was observed on AgBr colloids, but not on the gray and yellow silver colloids prepared by traditional means. The possible reason was explained. One major advantage of this means is the absence of the spectral interference such as citrate, BH4- arising from reaction products of the colloids formation process. On AgBr colloids, one can get some molecular SERS impossible to get on the gray and yellow silver colloids. PMID- 18422133 TI - [Vibrational spectra of scheelite and its color genesis]. AB - In order to study the spectral characteristics and color-genetic mechanism of the scheelite with orange color found in scheelite-tin-beryl (W-Sn-Be) deposits in Xuebaoding, four groups of scheelite were investigated by means of EMPA, IR, LRM and XRD. The scheelite samples were tested by XRD and IR in order to reveal if there are other minerals intermingled with scheelite. Meanwhile, the structure, the structure water and the crystal water were studied through infrared spectra. The absorption bands and peaks of infrared and Raman spectra were assigned. Finally, it seems that there is no distinct color difference between scheelite from Xue baoding and those from other places. So, it is conceived that the color causing may correlate with the minor crystal structure, trace element or REE in the crystal. PMID- 18422134 TI - [Spectroscopic characterization of binding between human serum albumin and 3,3', 4',7-tetrahydroxyflavone]. AB - The binding mechanism of human serum albumin with 3,3', 4', 7-tetradroxyflavone (FIS) was characterized by fluorescence and UV absorption spectra. The intrinsic fluorescence of HSA was significantly quenched by FIS in the physiological condition (pH 7.4), and the quenching mechanism is mainly static quenching process in the drug to protein molar ratio ranging from 0.1 to 10. The results revealed that the drug and protein formed 1 : 1 complex with the binding constants of (1.05 +/- 0.18) x 10(5) L x mol(-1). The dissociation behaviors of FIS in different pH conditions were investigated by UV absorption spectra, and it was found that FIS existed in the mixtures of ionic and neutral species. The UV absorption band I of FIS has a significant red shift (above 40 nm) after combination with HSA, demonstrating that FIS was bound to protein in ionic species which was driven by electrostatic force. The second derivative spectra of FIS showed that the combination included specific and non-specific forms. The intrinsic fluorescence of FIS was conspicuously enhanced in the presence of HSA due to the excited-state proton transfer (ESPT) and this further confirmed the complex formation of HSA with FIS. PMID- 18422135 TI - [Relation between complexes iotramolecular transfer and structure of [M(SS) (NN)](M = Zn2+, Cd2+)]. AB - The title complexes of [M(SS)(NN)](M = Zn2+, Cd+; SS = 1,2-dicyano 1,2-ethylene dithiolate or maleonitriledithiolate; NN = 5-nitro-1,10 phenanthroline) were synthesized. The electronic absorption spectra of complexes [M(SS)(NN)] in DMSO, DMF, acetone and Py were reported. The spectra absorption bands 270-280 nm, 320 350 nm and 350-390 nm are the intraligand pi(b) --> pi* transfer of ligand mnt(2 ) and 5-NOz-phen. The spectra absorption band 400-500 nm is intramolecular charge transfer (LL'CT) from a ligand (mnt(2-)) to another ligand (5-NO2- phen) The relation between electronic spectra absorption band and correlative transition in the molecular orbital energy level diagram of the complexes was investigated. The LL'CT band of the title complexes is weaker by comparison with M(SS) (NN) (M = Co2+, Fe2+, Ni2+ and Cu2+). The LL'CT band of [M(SS) (NN)](M = Zn2+, Cd2+) (7b2 to 7b1) is symmetry forbidden. PMID- 18422136 TI - [Study on the mechanism of color changes of Azur A and sodium dodecyl sulphate before micelle formation]. AB - The mechanism of color changes of sodium dodecyl sulphate(SDS) and Azur A (AA) before micelle formation was studied by spectral probe. The changes in the absorption spectra of SDS-AA and chondroitin sulfate(CS)-AA complexes were compared. The influence of ethanol and NaCl on the absorption spectra of SDS-AA complex was investigated. It was found that SDS-AA system showed color changes from blue to amaranth to blue due to the difference in the spatial orientation aggregative degree of AA binding on SDS regular aggregate. The critical concentration of premicelle formation (CPC) was found to be 1.0 x 10(-4) mol x L( 1) by the relation between c(SDS) and deltaA498/deltac(SDS). PMID- 18422137 TI - [Measurement and analysis of absorption spectrum of human blood]. AB - The present paper puts forward a method of disease diagnosis by using the technology of spectrum analysis of human blood serum. The generation mechanism of absorption spectrum is explained and the absorption spectra of the normal blood serum and the sick blood serum are listed from the experiments of absorption spectrometry. Though the value of absorbency of the sick blood serum is almost equal to that of the normal blood serum in the most absorption spectra, there are some differences around 278 nm in the absorption spectrum. The absorbency of the blood serum with hyperglycemia is greater than that of the normal blood serum at 285 nm in the spectrum, and besides, there comes a peak shift of absorption with hyperglycemia. In the absorption spectrum of the blood serum with hypercholesterolemia, there is a clear absorption peak at 414 nm. However there is not any peak at that wavelength in the absorption spectrum of the normal blood serum. Through comparing the characters of the spectrum, we can judge if the blood sample is or not, and this blood analysis is a new method for the diagnosis of disease. Compared with other methods of blood measurements, the method of absorption spectrum analysis of blood serum presented in this paper, is more convenient for measurement, simpler for analysis, and easier to popularize. PMID- 18422138 TI - [Determination of chlorophyll a and b using absorption spectrum]. AB - Chlorophyll a and chlorophyll b play a significant role in the plant growth process. Precise determination of its content could provide the scientific basis for the corps growth state, the plant pathology diagnosis and so on, and is the key point of implementing accurate agriculture. The authors transfered the absorption spectrum experiment data of chlorophyll a and chlorophyll b determined by predecessor into a transmission spectrum. Observing the transmission spectrum using human eyes, the color of chlorophyll a and chlorophyll b was obtained. Thus, according to the predecessor's experiment curve of absorption spectrum of chlorophyll a and chlorophyll b, combining the three primary color principles of colorimetry, and choosing a standard light source with spectrum power distribution similar to D65 as the illumination, the authors determined their chromaticity coordinates in the 1931CIE-x, y chromaticity diagram: Chlorophyll a is (0.198 1, 0.334 1), which falls in the cyan color region; Chlorophyll b is (0. 270 4, 0. 566 3), which falls in the yellowish green region. The main point of above processing is: reducing the spectrum curves of chlorophyll a and the chlorophyll b to coordinates on the chromaticity diagram, and the result could offer the essential theoretical support for a new non-contact, long-distance and non-damage technology to determine the pigment content in single leaf or mass. PMID- 18422139 TI - [Optical limiting cycling performance and photostability of vanadyl phthalocyanines and naphthalocyanines]. AB - Optical limiting (OL) cycling performance and photostability of vanadyl phthalocyanines (VOPcs) and naphthalocyanines (VONcs) were studied. The results show that although vanadyl 2,11,20,29-tetra-tert-butyl-2,3-naphthalocyanine has the most pronounced OL effect originally, it is not a promising OL material due to its poor photostability. Meanwhile, vanadyl 2,9, 16,23-etraphenoxy-29H, 31H phthalocyanine is considered to be a potential OL material, owing to its considerable OL effect and good photostability. A possible photochemical mechanism behind the reverse saturable absorption (RSA) and photolysis process of the studied species is proposed. PMID- 18422140 TI - [Study on temperature measurement and control for microfluidic systems]. AB - A technique for temperature control and measurement in microfluidic systems was developed. The technique is easy to be implemented in an analytical lab and the experimental setup is relatively simple and inexpensive. The parts for temperature measurement include a CCD camera, a fluorescence microscope and an image acquisition card. The spatial distribution and the temporal variation of temperature in microfluids were measured. The method is non-invasive and the spatial and temporal resolutions were 0. 8 microm and 40 ms, respectively a thermal control device was built using indium-tin-oxide (ITO) coated glass as heater, taking advantage of its transparent quality. The temperature could be controlled with a precision of +/-0.1 degrees C. The image processing procedure was optimized and the efficacy of the method was demonstrated. PMID- 18422141 TI - [Study on mechanism of lanthanide sensitized fluorescence energy transfer between terbium(III)-ciprofloxacin and rhodamine B]. AB - A complex composed of ciprofloxacin and terbium (Tb3+) in the solution of acetic acid-sodium acetate buffer (pH 6.2) was studied by fluorescence spectroscopy and ultra-violet absorption spectroscopy. The emission peak of Tb3+ appeared at 490, 545, 590 nm, (the sensitized fluorescence was enforced mightily) and the intensity of 545 nm emission peak was increased obviously. In its acute emission spectrum, the strongest emission peak of Tb3+ was at 545 nm, which was close to the wavelength of the biggest absorption peak of RB, 552 nm. Therefore, as the basic dye rhodamine B(RB) was added, the fluorescence intensity of 545 nm emission peak decreased regularly, indicating that there was a great quenching process. The result showed that the course was statistic. Based on the mechanism of the Forster energy transfer, the efficiency of energy transfer and the distance between the acceptor RB and the complex were found. Thereby, it was indicated that the course of action was single static quenching and the mechanism of quenching was based on energy transfer. PMID- 18422142 TI - [Fluorescence polarization used to investigate the cell membrane fluidity of Saccharomyces cerevisiae treated by pulsed electric field]. AB - To know the lethal mechanism of microorganisms under pulsed electric field treatment, the relationship between the inactivation of Saccharomyces cerevisiae (CICC1308) cell and the permeability and fluidity changes of its cell membrane treated by pulsed electric field (0-25 kV x cm(-1), 0-266 ms) was investigated. With 1,6-diphenyl-1,3,5-hexatriene (DPH) used as a probe, the cell membrane fluidity of Saccharomyces cerevisiae treated by pulsed electric field was expressed by fluorescence polarization. Results showed that the cell membrane fluidity decreases when the electric flied strength is up to 5 kV x cm(-1), and decreases with the increase in electric field strength and treatment time. The plate counting method and ultraviolet spectrophotometer were used to determine the cell viability and to investigate the cell membrane permeability, respectively, treated by pulsed electric field. Results showed that the lethal ratio and the content of protein and nucleic acid leaked from intracellular plasma increased with the increase in the electric field strength and the extension of treatment time. Even in a quite lower electric field of 5 kV x cm( 1) with a tiny microorganism lethal level, the increase in UV absorption value and the decrease in fluidity were significant. It was demonstrated that the cell membrane fluidity decreases with the increase in lethal ratio and cell membrane permeability. The viscosity of cell membrane increases with the decrease in fluidity. These phenomena indicated that cell membrane is one of the most key sites during the pulsed electric field treatment, and the increased membrane permeability and the decreased cell membrane fluidity contribute to the cell death. PMID- 18422143 TI - [Study on the interaction of serum albumin and micromolecule compounds by modified fluorescence spectra]. AB - The fluorescence spectroscopy is widely used in the analysis of the interaction between serum albumin (SA) and micromolecule compound, in which SA is measured as the fluorescence source to provide interaction information. A modified method has been compared with the classical method in the present paper, where the micromolecule compounds were used as the interested analytes whose fluorescence spectra were measured. The binding constants obtained based on the classical and modified methods were found much different by studying the two systems, BSA-TX and BSA-SH. The result shows that the classical method has the drawback of taking the fluorescence source of tryptophan instead of all other amino acids groups in SA molecule. Two novel methods were then reported, namely the modified fluorescence spectrometry method and fluorescence background subtracting method. The former could reflect the whole information of the interaction system, while the latter effectively over-came the overlay of spectra in the system, where the fluorescence spectra of the two components interfere with each other terribly. The effectivity of the two novel methods has been proved by ultra-violet spectrometry. PMID- 18422144 TI - [Study on application of multi-wavelength LED array induced fluorescence spectrum in multicomponent analysis]. AB - The technique of fluorescence spectrum is one of the efficient methods for monitoring and identifying dissolved organic matter, oil pollutants and biomass of phytoplankton in water. A new kind of fluorescence spectrum system with a multi-wavelength LED array as the exciting light source is introduced. The principle of the system and the method for multi-component analysis are discussed. The excitation-emission matrix (EEM) fluorescence spectra of samples mixed with different concentration fluorescent dyes were measured with this system in the authors' laboratory, which were analyzed using parallel factor (PARAFAC) algorithm. The results indicate that the correlation coefficient of the resolved concentration and actual ones is up to 98%, which shows the potential application of the fluorescence spectrum system in multi-component analysis. PMID- 18422145 TI - [Fluorescence spectra and quantum yield of TiO2 nanocrystals synthesized by alcohothermal method]. AB - Fluorescence spectra and fluorescence quantum yield of TiO2 nanocrystals were studied. Using tetra n-butyl titanate as a starting material, a facile alcohothermal technique was used to synthesize TiO2 nanocrystals. As can be seen from the transmittance electron microscopy (TEM) image, TiO2 nanocrystals with a relatively uniform particle size distribution of < 10 nm are present in the transparent sol. The transparent sol presents a strong stable fluorescence emission with a maximum at 450 nm, which is greatly dependent on the size quantization effects, defect energy level and the surface state of TiO2 nanocrystals. The quantum yield (gamma) of TiO2 was determined by the relative comparison procedure, using freshly prepared analytical purity quinine sulfate in 0.05 mol x L(-1) H2SO4 as a relative quantum yield standard. The emission quantum yield of TiO2 nanocrystals prepared in alcoholic media was calculated to be about 0.20 at wavelengths ranging from 330 to 370 nm, which was much higher than the values reported in previous works. So, it is supposed that nano-TiO2 will be applied as a potential quantum dots fluorescence probe in biological analysis. PMID- 18422146 TI - [Three-dimensional excitation emission matrix fluorescence spectroscopic characterization of dissolved organic matter in water of coal-mining area]. AB - Three-dimensional excitation emission matrix was applied to characterize the fluorescence properties of dissolved organic matter in various waters of Shilong coal-mining area. Fluorescence peak I (fulvic-like) and peak II (humic-like) were strong, while peak IV and peak V (protein-like) were weak or even undetected in some samples. Fluorescence peaks in various waters and different zones showed great difference in intensities and the fluorescence peaks in underground water tended to be much lower than those of surface waters. Furthermore, the fluorescence peaks of rivers and lakes were higher than those of mine drainage, and also the fluorescence peaks in coking zone and coal mining zone were higher than those in sewage-irrigated zone, or even much higher than those in farming zone. The reason may be that coal mining activities and coal industry can bring plenty of organic matter from coal to surroundings. Meanwhile, surface water would accept mine drainage, waste water of coal-washing and sewage from daily life easier than underground water, so surface water can be polluted seriously. Fluorescence peaks in waters from coal mining area are little influenced by pH of the water but can be influenced by the content of Ca2+ to water in some extent. PMID- 18422147 TI - [Study of 1-butanol absorption and fluorescence spectra induced by UV-light]. AB - The absorption and fluorescence spectra of 1-butanol solution induced by UV-light and its characteristics were studied experimentally and theoretically. The authors found that 1-butanol solution shows a good absorption in 190-250 nm, and fluorescence was detected when excited by 210-250 nm. Fluorescence spectra of the mixed solution with 1-butanol and n-hexane were measured, and the fluorescence intensity shows a linear relationship with the volume ratio of the former. For the mixed solution of 1-butanol and alcohol, the authors found that at different wavelength, as the matching of 1-butanol and alcohol solution changes, the law for the intensity of fluorescence also changes. Meanwhile, the mathematical formula for the fluorescence position of the mixed solution was listed, and the error between calculation and experiment has been proved to be small. This may provide some reference for the further research of fluorescence spectrum of alcohol resolutions. PMID- 18422148 TI - [Enhancing effect of ethanol additive on the plasma emission in ICP source]. AB - In the present paper, the effects of the ethanol-water solution temperature (20, 40, and 60 degrees C) on the emission intensity of ICP source, plasma parameters (excitation temperature and electron density), physical properties (surface tension and viscosity) of sample solution, and nebulization (uptake rate, effective uptake rate and nebulization efficiency) were studied by using optical to-electrical detection method. The experimental results showed that the velocity of the sample entering the plasma increased, so that the line intensity of the elements Zn, Fe, Mg, Si, and Sr in water increased obviously with increasing the content of ethanol and the rise of the corresponding solution temperature. And the excitation capacity in the plasma was strengthened with the rise of the ethanol-water solution temperature. The maximal spectrum intensity of the elements Zn, Fe, Mg, Si, and Sr at 60 degrees C was higher by 55.8%, 45.4%, 48.9%, 17.7%, and 21.6% than that at 20 degrees C, respectively. It's valuable for the detection of the trace element. PMID- 18422149 TI - [Application of ICP-MS to detection of mineral elements and heavy metals in transgenic cotton seeds]. AB - With the rapid development of transgenic cotton, people begin to focus on its physiology and ecology. The present paper studied the contents of heavy metals and microelements in transgenic cotton seeds by ICP-MS/ICP-AES. The results showed that the content of most microelements detected in transgenic cotton seeds, B, Na, Si, P, K, Ca, Mn, Co, Ni, Zn, Se and Mo was lower than that in regular cotton seeds, except Mg, Fe and Cu. And the content of most heavy metals detected in transgenic cotton seeds, Al, As, Cd, Sb, Tl, Pb and Bi, was higher than that in regular cotton seeds, except Cr and Hg. All the data showed that the accumulation of heavy metals and microelements in transgenic cotton seeds is disadvantageous to itself. PMID- 18422150 TI - [Determination of lead, cadmium, mercury, chromium and arsenic in acrylonitrile butadiene-styrene copolymer using microwave digestion-ICP-MS]. AB - A method was studied for the analysis of Cr, As, Cd, Hg and Pb in acrylonitrile butadiene-styrene copolymer by using ICP-MS. The instrument parameters were optimized and the introduction system was developed systematically. The sample is decomposed by microwave digestion. The digestion condition was optimized concerning digestion system, proportion of acids and digestion procedure, which affords reference for the preparation of the same kinds of polymer samples. The detection limits of the method for all sample elements were 0.7-6.5 ng x g(-1), the recoveries were 89.8%-110.8%, and the RSDs were 2.8%-11.3%. The analytical method presented was characterized with good precision and accuracy, simplicity, rapidness, low limits of detection and no matrix matching requirements. PMID- 18422152 TI - [Determination of Al and ca in soil by laser micro-plasma spectroscopy]. AB - Laser micro-spectral analyzer, coupled with CCD spectrometer, was used to determine soil elements in argon atmosphere at reduced pressure in the authors' experiment. The accuracy and availability of this method were studied. With Al I 394.40 nm and Ca II 396.85 nm as spectral analysis lines, using "the three standard samples method", the contents of Al and Ca in soil were determined. The results show that the maximum of relative standard deviation (RSD) of quantitative analysis is 5.80%; the maximum of relative difference between the quantitative analysis result and the standard value is 7.65%, suggesting that the accuracy of determination of Al and Ca in soil meets the challenge of quantitative analysis. PMID- 18422151 TI - [Determination of Si, Al and other in biological samples by ICP-AES with fusion with LiBO2 after dry ashing]. AB - An alkaline sample digestion procedure using fusion with lithium metaborate (LiBO2) after dry ashing was developed. Silicon, aluminium and other, a total of 10 elements in different kinds of biological samples, were measured by ICP-AES. The detection limit of this method was 0.15 microg x mL(-1) for Si and 0.03 microg x mL(-1) for Al. The accuracy and precision of the method were demonstrated by analyzing certified reference materials poplar leaves (GBW07604). The precision was less than 3% and the accuracy was -0.03% for Si and -3% for Al. This method was applied to the certification of a series of ten biological reference materials. The results were satisfied. PMID- 18422153 TI - [Comparative study on trace elements in natural colored cotton and white cotton]. AB - A method was studied for simultaneous determination of multi-elements in natural colored cotton by microwave digestion and ICP-AES. The contents of Ca, Mg, Fe, Cu, Ba, Zn, Al, Sr, Mn and Si in colored and white cotton were determined by this method. The recovery ratio obtained by standard addition method ranged between 93% and 111%, and the relative standard deviation was below 4% (n = 5). The results showed that the contents of Ca, Cu, Zn, Al, Fe, Sr and Si in green and brown cotton are higher than those in white cotton. The data from the study gives some references for further researches on the colored cotton. PMID- 18422154 TI - [Application of IC-HG-AFS determination method for arsenic in gold mine tailings]. AB - A method for the determination of species (arsenite (As(III)), arsenate(As(V)), monomethylarsonic acid (MMA) and dimethylarsinic acid (DMA)) in solution samples of gold mine tailings has been applied. The analytical method used was ion chromatography coupled on-line to atomic fluorescence spectroscopy through hydride generation with the mobile phase (NH4)2 HPO4 (pH 6. 0). The recovery with the standard solution of arsenic added into the water samples was very good (recovery 95%-109%). The arsenic species were separated completely; arsenic speciation was stable and the detection limits were very low: 1.0 microg x L(-1) As(III) and 2.3 microg x I(-1) for As(V). The method is fit for the determination of arsenic species in the solution of gold mine, gold mine tailings and the polluted water around the gold mine. PMID- 18422155 TI - [Indirect study of zinc-cystine complex reaction by FAAS]. AB - Each state of cystine and zinc ion in aqueous solution under different pH conditions was calculated by computer, and the scatter diagram was given. Under the influence of solution pH, the mechanism of complex reaction in indirectly determining cystine by flame atomic absorption spectrometry with ZnS was studied. The soluble complex ion is composed of 0 valence cystine Cys-Cys+/-, -1 valence cystine Cys-Cys+/- and -2 valence cystine Cys-Cys(2-) with Zn(OH)2. The theoretical analysis from computing and scatter diagram dovetail very well with the data determined from the experiments. The structure of soluble zinc-cystine complex ion is, [(COO-) CH (NH3+) CH2S-SCH2 CH (NH3+) COO-] Zn (OH)2, [(COO-) CH (NH3+) CH2S-SCH2CH(NH2)COO-]Zn(OH)2 and [(COO-)CH(NH2)CH2S-SCH2CH(NH2)COO ]Zn(OH)2. PMID- 18422156 TI - [Novel ionic liquid as solvent for preconcentration of trace lead in dialysis fluids and its determination by graphite furnace atomic absorption spectrometry]. AB - A novel room temperature ionic liquid 1-butyl-3-trimethylsilylimidazolium hexafluorophosphate was designed and synthesized, and was applied to the preconcentration of ultra trace lead in a relatively large volume of dialysis fluids (such as 1 000 mL or bigger). In the present work, dithizone was employed as chelator to form a neutral lead- dithizone complex, and the ionic liquid 1 butyl-3-trimethylsilylimidazolium hexafluorophosphate was used to displace CCl4 as solvent for liquid/liquid extraction of lead complex. The ionic liquid was collected and the lead (II) was back-extracted into aqueous solution by adding nitric acid solution, and the above solution was directly applied to determine lead in the dialysis fluids by graphite furnace atomic absorption spectrometry. Experiments indicated that the proposed extraction system is superior over other systems in which organic solvent such as CCl4 or classical room temperature ionic liquid1-butyl-3-methylimidazolium hexafluorophosphate was selected as extraction medium, and its extraction efficiency of once extraction and enrichment factor of lead was more than 99% and 200 times, respectively. The preconcentration combined with graphite furnace atomic absorption spectrometry has been applied to the determination of trace lead in dialysis fluid samples with satisfactory results. PMID- 18422157 TI - [Study on adsorption behavior of nanosized barium-strontium titanate powder for lead ion in water using FAAS]. AB - Nanosized barium-stroutium titanate (BST) powder, a double salt sorbent, was prepared from BaCl2, SrCl2 and TiCl4 by using oxalate co-precipitate method, and characterized by using transmission electron microscope (TEM), X-ray diffraction (XRD) and Fourier transform infrared spectrophotometer (FTIR). By means of determination with flame atomic absorption spectrometry (FAAS), the adsorption characteristic of lead on the BST powder w as investigated. The results showed that theforms of the BST powders are clubbed, highly pure perovskite, and the average particle diameter is 36 nm. Lead can be adsorbed strongly on nanosized BST. The adsorption capacity of BST nanoparticle towards lead was found to be 13 mg x g(-1) when the pH is 6.0, and 0.5 mol x L(-1) HNO3 is sufficient for complete elution. A new method for the determination of trace lead based on BST nanoparticle separation/preconcentration and FAAS determination was proposed. The detection limit of this method for lead ion is 11 microg x L(-1), and the relative standard deviation is 2.6%. The method has been applied to the determination of trace lead in water samples with satisfactory results. PMID- 18422158 TI - [Digestion-flame atomic absorption spectroscopy]. AB - A microwave digestion-flame atomic absorption spectroscopy (FAAS) method was developed for the determination of metal elements Na, Zn, Cu, Fe, Mn, Ca and Mg in Mongolian patents. The instrument parameters for the determination were optimized, and the appropriate digestion solvent was selected. The recovery of the method was between 95.8% and 104.3%, and the RSD was between 1.6% and 4.2%. The accuracy and precision of the method was tested by comparing the values obtained from the determination of the standard sample, bush twigs and leaves (GSV-1) by this method with the reference values of GSV-1. The determination results were found to be basically consistent with the reference values. The microwave digestion technique was applied to process the samples, and the experimental results showed that compared to the traditional wet method, the present method has the merits of simplicity, saving agents, rapidness, and non polluting. The method was accurate and reliable, and could be used to determine the contents of seven kinds of metal elements in mongolian patents. PMID- 18422159 TI - [Analysis and study of total flavone and trace element in carthmus tinctorius L]. AB - The content of total flavone in carthmus tinctorius L in Xinjiang was determined by ultraviolet spectrophotometry with scan from 200 to 400 nm where the maximum absorption wavelength was 267.00 nm. The contents of ten trace elements and constant elements in carthmus tinctorius L including potassium, sodium, calcium, magnesium, iron, chromium, nickel, manganese, copper and zinc were determined by flame atomic absorption spectrophotometry. The results showed that there were comparatively rich trace elements and high total flavone, among which are comparatively high calcium, magnesium, chromium, copper and zinc in carthmus tinctorius L. It provided useful data for discussing the relationship between the content of total flavone and the trace elements in carthmus tinctorius L, and for studing the relationship between the trace elements in carthmus tinctorius L and the cure for cardio-cerebral vascular disease. PMID- 18422160 TI - [Determination of microelements in C. tangutica (Maxim.) korsh flower by FAAS]. AB - The powder was digested with HNO3-HClO4 mixed acid (4 : 1) before determination. The eleven microelements in C. tangutica (Maxim.) korsh flower were determined by FAAS, and the determination medium was 4% nitric acid. The work conditions, accuracy and precision of the method were studied. The experimental results show that the interaction of the elements were small under the conditions of determination, and no significant effect on the result. The linear correlations of standard curves are good (r = 0.9871-1.000 0). The recovery (n = 7) is 98.3% 105.1%, and the RSD (n = 7) is 0. 23%-1.07%. It can be used to determine the trace elements simultaneously in C. tangutica (Maxim.) korsh flower. The method has good precision and accuracy, so it is able to meet the requirements for analysis. The contents of Ca, Mg, K, Cu, Fe, Zn, Mn, Na and Co were 206.30, 284.50, 3415.20, 0.116 6, 62.171, 3.275, 67.826 5, 28.00, 0.133 3 mg (100 g)( 1)', respectively, while Cd and Ni were not found in the samples. The results provided theoretical basis for the effect of C. tangutica (Maxim.) korsh herbal medicine. PMID- 18422161 TI - [Study on applications of heteropoly acids (blue) spectrophotometry to determination of trace elements]. AB - Heteropoly acids can be reduced to the pigmented heteropoly blue. Therefore a reliable method for determination of trace elements can be founded by association with spectrophotometry. At present, heteropoly acids spectrophotometry has made great progress in trace analytical field for its high accuracy, simple instruments and rapidity in detection. In the present paper, the application and research progress of heteropoly acids in the determination of trace elements (P, As, Si and V) are reviewed. The methods include the molybdenum blue method, the heteropoly-basic dye approach, fluorophotometry, the flow injection analysis, atomic absorption spectrophotometry, laser thermal spectrophotometry, etc. Finally, analysis and comparison of these methods are studied. PMID- 18422162 TI - [Redshift estimation of galaxy spectra based on similarity measure]. AB - Automated spectra analysis is desirable and necessary for efficiency of large sky surveys such as SDSS (Sloan digital sky survey), 2DF (2 degree fields) and LAMOST (large sky area multi-object spectroscopic telescope). In the present paper, we present a method for redshift estimation of galaxy spectra based on similarity measure. Firstly, we extract the spectral lines of the observed spectrum using the feature constrains of spectral lines; secondly, the authors determine the redshift candidates of the observed spectrum by spectral line features; then, the similarity between the observed spectrum and the template spectra shifted by each redshift candidate is measured; finally, the candidate of the highest similarity is chosen as the estimated redshift. PCA (principal component analysis) is used to build the static galaxy template spectra. The authors perform PCA for the four template spectra E, S0, Sa and Sb of the normal galaxy and the seven template spectra Sc, Sb1, Sb2, Sb3, Sb4, Sb5 and Sb6 of the starburst galaxy respectively, where the eleven template spectra are presented by Kinney & Calzetti et al. Two eigen-spectra are produced with the variance contribution rate of 99%. The authors choose the two eigen-spectra as the galaxy templates. The similarity measure proposed, which is similar to the evidence accumulation, is defined as the weighted sum of several similarity evidences. It can reduce the influence caused by some error matching. The authors divide the observed spectrum and the template spectrum respectively into several parts, and measure the correlations of the corresponding parts of them, which is chosen as the similarity evidences in the proposed similarity measure. The principle of setting the weights is that the higher the correlation, the higher the corresponding weight. The proposed approach is compared with the method based on spectral line matching and the traditional cross correlation technique by experiments, the results show that the proposed method has a higher correct rate. PMID- 18422163 TI - [Choice of treatment plan and operation time on pediatric chronic sinusitis]. AB - OBJECTIVE: To explore the curative effect of pediatric chronic sinusitis and analyze its relative factors, so that the best treatment plan and operation time can be choose. METHOD: Two hundred and twenty cases of pediatric chronic sinusitis were divided into 4 groups: group A with only simple chronic sinusitis, group B accompanied adenoid hypertrophy. Among group B, group B1 with no nasal cavity structural abnormalities but group B2 on the contrary. Group C accompanied only nasal cavity structural abnormalities. According to the documents, all patients were administered with standard and systematical three-stage treatment. The curative effect of each group was calculated and analyzed in each treated stage. RESULT: After the first stage treated in 220 pediatric chronic sinusitis, a statistical significance of curative effect could be observed between group A and B, A and B1, A and B2, A and C (P < 0.01), but none between group B and C, B1 and B2, B1 and C, B2 and C (P > 0.01). Ninety-four patients in group B were accepted the second stage treated: adenoidectomy after first treatment failed. There was a statistical significance of curative effect between group B1 and B2 (P < 0.01). Once the above management both failed, endoscopic sinus surgery (FESS) was carried out. The total effective rate of FESS was 89.65%. CONCLUSION: The systemic, disciplinary medical management has some referred value in the treatment for children with chronic sinusitis. Adenoid hypertrophy and nasal cavity structural abnormalities are two relative factors of the curative effect in pediatric chronic sinusitis. The time and cost of therapy can be saved if adenoidectomy and FESS are done for cases with adenoid hypertrophy or (and) nasal cavity structural abnormalities with the systemic, disciplinary medical management simultaneously. PMID- 18422164 TI - [Effects of long term use of beclomethasone dipropionate nasal spray on bone density with perennial allergic rhinitis]. AB - OBJECTIVE: To study the effects of long term use of beclomethasone dipropionate (BDP) nasal spray on bone density with perennial allergic rhinitis (AR) in adults. METHOD: A 5-year randomized study was conducted on the effects of BDP nasal spray on serum calcium, phosphorus, alkaline phosphatase, and bone density determined before and after the treatment in 36 adult patients with perennial AR. 20-45 years of age, were randomly divided into 3 groups. That is group A (nasal spray 1 - <3 year), group B (nasal spray BDP 3 - <5 year) and group C (nasal spray BDP > or =5 year). The data were analyzed by paired t test. RESULT: The perennial AR were followed up for more than > or =1 year, > or =3 year and > or =5 year to observe the influences of nasal spray BDP. There were no significant difference between the data examined before and after the treatment (P > 0.05). Bone development is not influenced by nasal spray BDP < or =400 microg/d within 5 years. CONCLUSION: Long term use of BDP nasal spray in adult patients does not lead to osteoporosis if the lowest effective steroid dose is given. PMID- 18422165 TI - [Effects of intranasal budesonide on the expression of c-fos and c-myc in nasal polyps]. AB - OBJECTIVE: To investigate the effect of intranasal glucocorticoids treatment on the expression of c-fos and c-myc nasal polyps. METHOD: Immunohistochemistry method was used to determine c-fos and c-myc expression in nasal polyps from patients with topical steroids treatment for 10-12 weeks and untreated patients. RESULT: The rate of c-fos expressing cases was 15% and the rate of c-myc expressing cases was 20% in nasal polyps from topical steroid treated patients, but in untreated patients, the rate of c-fos expressing cases was 80% and the rate of c-myc expressing cases was 85%. There were significant differences between two groups (P < 0.01). The c-fos and c-myc expression was remarkably downregulated in nasal polyps from topical steroid treated patients compared to untreated patients. CONCLUSION: The results show that glucocorticoids may induce cell apoptosis in nasal polyps by depressing the c-fos and c-myc expression. PMID- 18422166 TI - [Expression and significance of nuclear factor-kappaB and ICAM-1 mRNA in nasal mucosa of allergic rhinitis]. AB - OBJECTIVE: To clarify the pathogenesis of allergic rhinitis (AR) by detecting the activation of NF-kappaB and the expression of ICAM-1 mRNA in nasal mucosa of the patients with AR. METHOD: AR patients were collected as experimental group and deflection of nasal septum patients as control group. Immunohistochemistry methods were used to examine the expression of NF-kappaB p50 and p65 subunits in the middle turbinate mucous tunic. ICAM-1 mRNA were detected by means of RT-PCR. RESULT: NF-kappaB was strongly expressed in AR group. NF-kappaB p50 existed in cytoplasm and nucleus. NF-kappaB p65 was mainly expressed in cytoplasm and less in nucleus. While NF-kappaB was expressed little in control group. The difference of NF-kappaB p50 and NF-kappaB p65 expression between two groups were significantly (P < 0.01). ICAM-1 mRNA was more strongly expressed in AR group (P < 0.01) while feebly expressed in deflection of nasal septum group. The expression of ICAM-1 mRNA were significantly correlated with that of NF-kappaB p50 and NF-kappaB p65 (r = 0.899 5, P < 0.01; r = 0.7601, P < 0.01). CONCLUSION: NF-kappaB plays a key role in AR. The excessively activated NF-kappaB promotes the transcription of ICAM-1 mRNA. ICAM-1 relates to the pathogenesis and development of AR. PMID- 18422167 TI - [Nasal sinus mucoceles after sinus surgery or trauma]. AB - OBJECTIVE: To analyse the causes of nasal sinus mucoceles after sinus surgery or trauma and explore operative therapy and methods of avoiding mucoceles recurrence. METHOD: Retrospective analysis were made in 22 cases of nasal sinus mucoceles after sinus surgery or trauma. RESULT: All the cases underwent endoscopic mucosectomy. There is one recurrent frontal mucocele in follow-up one year later after surgery and coherency of the lateral nasal cavity with the middle nasal concha in two cases. CONCLUSION: Sinus surgery or trauma can induce nasal sinus mucoceles. So it is necessary to fix the sinus fracture site and replace the mucosa as possible, keep the the middle nasal concha stability and healthy sinus mucosa in endoscopic surgery,and take long term follow-up for reducing mucoceles recurrence. PMID- 18422168 TI - [Effect of allergic factors on chronic rhinosinusitis]. AB - OBJECTIVE: To explore the correlation between allergic factors and chronic rhinosinusitis, and discuss its effects on the development of illness. METHOD: One hundred and one cases were selected to detect allergic serum relative index: the concentration of serum TIgE, specific IgE semi-quantitative test, the concentration of serum ECP as well as inquiring medical record, Lund-Mackay CT system scoring. RESULT: The concentration of serum TIgE and serum ECP in CRS was higher than those in control (P < 0.01). The increased proportion of serum TIgE and serum ECP was significantly different among clinical stages (P < 0.05) and between the cases with and without prior sinus surgery (P < 0.05). CT scores of the cases with serum TIgE and serum ECP increased were significantly higher than those without serum TIgE and serum ECP increased. The strong positive rate in CRS was significantly different among clinical stages (P < 0.05). The strong positive rate increased significantly in the cases with prior sinus surgery (P < 0.05). CT scores in the cases with strong positive allergen were significantly higher than that in the cases without positive allergen (P < 0.05). CONCLUSION: These results suggest that allergic factors make some negative effect on the severity of CRS and are the index for the severity and worse prognosis of illness. PMID- 18422169 TI - [Study on 34 cases of nasal button battery]. AB - OBJECTIVE: To prevent the perforation of nasal septum due to button battery lodging in nose. METHOD: In 34 cases of nasal button battery, 28 were removed under general anesthesia and 6 under local anesthesia. The general treatment of nasal douching and local dressing were done after operation. RESULT: Only 1 case of perforation were found in 25 cases undertaken general treatment after operation, and perforation were found in all 9 cases with no general treatment. CONCLUSION: Nasal douching and local dressing after battery removing can alleviate the mucosal erosion and decrease the sequels of nasal button battery. PMID- 18422170 TI - [Analysis the causes and treatment of postnasal drip syndrome]. AB - OBJECTIVE: To analysis the causes and treatment for postnasal drip syndrome. METHOD: One hundred patients were tested by routine examination of otolaryngology, nasal endoscopy, fiberoptic laryngoscopy and nasal sinuses CT. Choice allergen test and nasopharynx X-ray were taken based on the symptoms and signs. Patients were treated accordingly for different causes, such as flushing the nasal cavity by herbal liquid, using glucocorticoid locally, taking medicine of improving the sticky film cilium function, immunotherapeutic or operation. RESULT: The causes included chronic rhinitis 22 cases (22%), chronic sinusitis or nasal polyp 31 cases (31%), allergic rhinitis 28 cases (28%), adenoid vegetation 16 cases (16%), chronic nasopharyngitis 3 cases (3%). All patients were treated accordingly and total cure rate was 82% after 6 months follow up. CONCLUSION: The causes of postnasal drip syndrome are complicated. It is critical to check the cause and chose adaptable therapy for good result. PMID- 18422171 TI - [Clinical analysis of solitary sphenoid sinus disease]. AB - OBJECTIVE: To explore the clinical characters and treatment on solitary sphenoidal sinus disease thus improve the initial diagnosis accuracy and curative rate together with avoiding severe complications. METHOD: Forty-six cases were included in the study. The clinical manifestations, medical imaging characteristics and type of pathology and surgical treatment efficacy were analyzed retrospectively. RESULT: The main symptoms were headache and visual lesion. Medical image showed pace occupying lesion and partial destruction of bone. Pathological exams showed 8 cases with sphenoiditis, 19 cases with fungal infection, 10 cases with cysts and 9 cases with polyposis. All the patients underwent open sphenoidal surgery with nasal endoscope. After a follow-up of over 6 months, most of the patients' symptoms had been obviously relieved. And the entrance to sphenoid was patent with smooth sinus wall and no secretion in sinus cavity. CONCLUSION: There is no specificity with solitary sphenoidal sinus disease. Pre-operation diagnosis rate can be improved with the help of nasal endoscope and CT or MRI. Nasal sinus surgery with nasal endoscope may be the first choice for treatment. PMID- 18422172 TI - [Detection of ADAR1 mRNA expression in larynx carcinoma tissues]. AB - OBJECTIVE: To investigate the expressions of RNA-dependent adenosine deaminase 1(ADAR1) mRNA in larynx carcinoma tissues, and to discuss its value in the development of larynx carcinoma. METHOD: The expression of ADAR1 mRNA in 51 larynx carcinoma and peri-carcinoma tissues were detected by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR). RESULT: ADAR1 mRNA was expressed broadly; the relative intensities of its expression in larynx carcinoma, larynx peri-carcinoma samples and larynx non-carcinoma tissue samples were respectively 2.963 +/- 0.912, 0.791 +/- 0.197 and 0.910 +/- 0.311. There were remarkable difference between larynx carcinoma and larynx peri-carcinoma, larynx carcinoma and non-carcinoma tissues. CONCLUSION: ADAR1 mRNA is expressed broadly in larynx carcinoma and may be play an important role in the development of larynx carcinoma. PMID- 18422173 TI - [Endoscopic removal of ethmoid osteoma: report of 13 cases]. AB - OBJECTIVE: To report 13 cases of ethmoid osteoma removed through an endoscopic approach and discuss the operative technique and clinical value. METHOD: Thirteen cases with ethmoid osteoma were retrospectively reviewed in the study. All procedures were performed with CT image guidance-according to the site of attachment. The anterior ethmoid cell was resected, and the whitish osteoma found. A double blunt elevator was used to separate the osteoma from ethmoid cell, and to push the osteoma medially toward the nasal septum, and (or) inferiorly toward the middle meatus. This method was performed in 7 patients (8 sides), whose CT scan showed the osteoma bases were not attached to lamina papyracea, and (or) anterior skull base. The others were drilled out with a intranasal powered drill for its attached to lamina papyracea in 4 patients and to anterior skull base in 2 patients. RESULT: Thirteen cases of ethmoid osteoma were removed completely through an endoscopic approach and there were no intraorbital or intracranial complication. Endoscopy showed normal epithelization at around 6 to 8 weeks after surgery. CONCLUSION: Preoperative CT scans, including coronal and axial images, is very helpful to determine the sites of the osteoma attachment and plan the operation. Endoscopic removal of ethmoid osteoma is not only an ideal, less invasive surgical approach, but also no cosmetic alteration. PMID- 18422174 TI - [Draf's endonasal drainage for inflammatory frontal sinus disease]. AB - OBJECTIVE: To introduce endonasal drainage procedures to frontal sinus in inflammatory sinus disease and its indications, methods and efficacy. METHOD: One hundred thirty two patients undergoing Draf I-III frontal sinus drainage procedures with 1-12 years follow-up were reviewed retrospectively. RESULT: Forty two patients underwent type I frontal sinusotomy, 43 type II sinusotomy and 47 type III sinusotomy. A successful result was seen in these groups, 83.4%, 83.7%, and 89.4% respectively. Best effect was gained by type III sinusotomy. There was no significant difference in efficacy between the different Draf frontal sinus drainage procedures (P > 0.05). CONCLUSION: Endonasal microscopic-endoscopic frontal drainage treatment of refractory, polypoid and recurrent frontal sinusitis can yield successful results. PMID- 18422175 TI - [Study of the relationship between obstructive sleep apnea syndrome and obesity and serum resistin level]. AB - OBJECTIVE: To investigate the serum resistin level in healthy volunteers and patients with obstructive sleep apnea syndrome (OSAHS), and study the relationship between OSAHS and obesity, serum resistin level. METHOD: All body's height, weight and calculate body mass index (BMI) were precisely measured. According to the BMI, 35 OSAHS patient were divided into two groups: One group was normal weight patients (BMI:18.5-24.9, 14 cases)and the other group was overweight patients(BMI > or =25, 21 cases). There were 40 healthy volunteers in control group. The blood pressure, fasting levels of plasma glucose (FPG), total cholesterol (TC) and triglyceride (TG), and test resistin level in plasma were detected. The apnea-hypopnea index (AHI), the average oxygen saturation and the minimum oxygen saturation in 35 OSAHS patients were examined with Polysomnography. RESULT: (1) Compared with healthy volunteers, serum resistin level, BMI, TG, systolic blood pressure (SBP) and diastolic blood pressure (DBP) in OSAHS patients were all enhanced (P < 0.05); (2) In OSAHS patients, the indexes of BMI, SBP, DBP and AHI for overweight group were notably higher than those for normal weight group (P < 0.05). (3) In OSAHS patients, BMI was positively correlated with AHI and SBP, and negatively correlated with the average oxygen saturation and the minimum oxygen saturation. (4) Serum resistin level was positively correlated with TG, FPG and DBP. Multiple stepwise regression analysis revealed that serum resistin level was associated with TG and FPG. CONCLUSION: In OSAHS patients, serum resistin level is enhanced and accompany with the disorder of blood pressure and serum lipid. Obesity or abdomen obesity may aggravate the hypoxia in OSAHS patients. Serum resistin level is positively correlated with TG, FPG and DBP. TG and FPG are important factors in serum resistin level, which indicate that the resistin is highly associated with lipid metabolism and glycometabolism, and possibly connect with diastolic blood pressure (DBP). PMID- 18422176 TI - [Acute injury of inhalable particle matters on nasal mucous membrane of rat]. AB - OBJECTIVE: To study the mechanism of the nasal mucous membrane inflammation induced by the inhalable particle matter (PM10). METHOD: Three dosage PM10 were instilled in rat nasal cavity of different groups for one week. The morphology of nasal mucosa and the numbers of inflammatory cell were observed in each samples. RESULT: The total numbers of inflammatory cells in PM10-treated groups were increased in a dose-respondent manner and significantly different from that in control group. The results of histopathological and scanning electron microscope (SEM) analysis indicated that PM10 caused nasal mucosa injury and pathological changes, such as the damage of cilia and nasal mucosa epithelium in a dose dependent way. The infiltration of inflammatory cells in nasal mucosa epithelium matrix, especially eosinophilia were observed. CONCLUSION: PM10 can cause rat's nasal mucosa inflammation and epithelial injury. PMID- 18422177 TI - [Comparison of content of 2,3,5,4'-tetrahydroxystilbene-2-O-beta-D-glucoside and anthraquinon in different ploidy Radix Polygoni Multiflori]. AB - OBJECTIVE: To investigate the content of 2,3,5,4'-tetrahydroxystilbene-2-O-beta-D glucoside and anthraquinon in diploid and triploid Radix Polygoni Multiflori. METHODS: 4 batches of Radix Polygoni Multiflori were collected from different districts. The content of 2,3,5,4'-tetrahydroxystilbene-2-O-beta-D-glucoside and anthraquinon in these samples were determined at 320 nm and 254nm wave length by HPLC with Inertsil ODS-3 C18 pillar and acetomitrile: aqua (25:75), methanol: 0.1% phosphoric (85:15) respectively as the mobile phase. RESULTS: The maximum content of 2,3,5,4'-tetrahydroxystilbene-2-0-beta-D-glucoside was diploid Radix Polygoni Multiflori from Deqing Guangdong. The maximum ratio of total anthraquinon was triploid Radix Polygoni Multiflori from Jinxi Guangxi reached 85%. CONCLUSION: The content of anthraquinon varies greatly in the samples from the different producing areas. PMID- 18422178 TI - [Study of quality on rhubarb from different habitats in Qinghai Province]. AB - OBJECTIVE: To investigate the effect of different habitats on rhubarb quality. METHOD: The rhubarb samples from various parts of Qinghai province were analysed by fingerprint. RESULTS AND CONCLUSION: The distribution and quality of wild Tangute rhubarb is better than Palmate sorrel rhubarb. The quality of high altitude sampes are better than low altitudes. The quality which from the grassland of plateau is better than chestnut soil area's, and the wild is better than that of planting. PMID- 18422179 TI - [Comparison of pharmacokinetics of tetrahydropalmatine monomer and extractive of corydalis and corydalis processed with vinegar]. AB - OBJECTIVE: To investigate the pharmacokinetics of tetrahydropalmatine monomer and extractive of corydalis and corydalis processed with vinegar in rats. METHODS: The plasma concentrations of tetrahydropalmatine were determined by a reversed phase high performance liquid chromatographic. The plasma concentrations-time data were calculated with 3p97 program. RESULTS: The main pharmacokinetic parameters of tetrahydropalmatine monomer and extractive of corydalis and corydalis processesd with vinegar as follows: T(1/2) was (5.66 +/- 1.92), (4.24 +/- l1.54), (4.35 +/- 1.34) h, Tmax was (1.5 +/- 0.5), (1.0 +/- 0.55), (0.5 +/- 0.68) h, Cmax was (0.71 +/- 0.29), (0.37 +/- 0.11), (0.67 +/- 0.35) microg/ml,AUC(o-t), was (2.58 +/- 0.85), (1.96 +/- 0.69), (2.95 +/- 1.61) microg/ml. CONCLUSION: The T(1/2) of corydalis and corydalis processesd with vinegar is more shorter than tetrahydropalmatine monome, the effect of preventing pain is promoted in rats. PMID- 18422180 TI - [The quality control of carbonized cortex moutan]. AB - OBJECTIVE: To set up method for the quality control of carbonized Cortex Moutan. METHODS: The optimized processing technology of carbonized Cortex Moutan was selected by the time of blood coagulation. Besides, the contents of tannin, adsorbability, paeonol and peoniflorin were researched. RESULTS: To parch the pieces 5 minutes at the temperature of 22 degrees C was the optimized processing technology. CONCLUSION: This study provides a reference to the institution for the quality standard of carbonized Cortex Moutan. PMID- 18422181 TI - [New chemical constituents from Radix Polygoni Multiflori after processing]. AB - OBJECTIVE: To study the new chemical constituents from Radix Polygoni Multiflori after processing. METHODS: Various kinds of chromatographic methods were used to deparate the chemical constituents from Radix Polygoni Multiflori after processing. Their structures were determined by NMR and MS spectral data. RESULTS: The two new compounds were 2,3-dihydro-3,5-dihydroxy-6-meth-yl-4 (H) pyran-4-one(I) and 5-hydruoxymethyl-furfuran(II). CONCLUSION: It is the first time that compound I and 1I were isolated from Polygoni. PMID- 18422182 TI - [Identification on Geranium by X-ray diffraction fourier fingerprint patterns and similarity degree of different fingerprint patterns]. AB - OBJECTIVE: To provide a X-ray diffraction (XRD) new method to identify different of herbal drugs of Geranium. METHODS: Analyse X-ray diffraction Fourier pattern of 1 plants of Geranium, 7 kinds substitute, and degree of different fingerprint patterns was calculated and analyzed according to the position (2 theta value) of peaks searched. RESULTS: Obtain the different X-ray diffraction Fourier pattern of 1 kinds plants of Geranium and 7 kinds substitutes of Geranium. CONCLUSION: This method can be used for indentification on different medicinal material of Geranium. PMID- 18422183 TI - [Studies on identification of gypsum fibrosum by X-ray]. AB - OBJECTIVE: To establish a new identification and analysis method of gypsum fibrosum, gypsum fibrosum praeparatum, pulvis talci and gypsum fibrosum blended with pulvis talci. METHODS: Powder X-ray diffraction fingerprint spectra method. RESULTS: Experiments and analysis were carried out on four samples. The standard X-ray diffraction fingerprint spectra and characteristic diffraction peaks of each sample were obtained. CONCLUSION: The experimental result indicates that X ray diffraction fingerprint spectra can be used to identify and analyze gypsum fibrosum, gypsum fibrosum praeparatum, pulvis talci and gypsum fibrosum blended with pulvis talci. PMID- 18422184 TI - [Headspace solid-phase microextraction-gas chromatography-mass spectrometry for analysis of volatile components from pollen pini]. AB - Headspace solid-phase microextraction HS-SPME) technique was employed to extract the volatile compounds from Pollen Pini. The volatile components were isolated and identified successfully by gas chromatography-mass spectrometry (GC-MS). The volatiles were collected using several SPME parameters (including extraction time and temperature). The best response was obtained using a PA fiber by when the extraction temperature was 70 degrees C, extraction time was 30 minutes and desorption time was 10 time. Analysis was performed by GC-MS. Seventy components were identified by HS-SPME. The main components were fatty compounds. The results showed that the HS-SPME technique could be used to extract the volatile components from HS-SPME-GC-MS. PMID- 18422185 TI - [The metabolites of cyclic peptides from three endophytic mangrove fungi]. AB - Nine secondary metaboites of cyclic peptide were isolated from three mangrove endophytic fungi Paecilomyces sp. (treel-7), 4557,ZZF65. They were viscumamide(1),cyclo(Pro-Iso)(2),cyclo(Phe-Gly)(3),cyclo(Phe-Ana)(4),cyclo(Gly Pro) (5),cyclo(Gly-Leu)(6), cyclo(Trp-Ana)(7),neoechinulin A(8),cyclo(Pro Thr)(9). The compounds 1,7,8,9 were firstly isolated from marine fungus. PMID- 18422186 TI - [Protection of vascular endothelial cells from ox-LDL induced injury by protocatechualdehyde]. AB - OBJECTIVE: To observe the protective effects of protocatechualdehyde on the human umbilical vein endothelial cells (CRL-1730) induced injury by ox-LDL. METHODS: The CRL-1730 were induced injury by ox-LDL, and then treated with protocatechualdehydes for 24 hours. The cytoactive of CRL-1730 was assessed by colorimetry of MTr, the NO level and NOS activity in the cell culture fluid were observed by colorimetry, and the expression of CD40 protein was determined by flow cytometry. RESULTS: Compared with the ox-LDL group, protocatechualdehyde increased,the number of CRL-1730 and the level of NO and NOS in cell culture fluid. Besides, protocatechualdehyde decreased the expression of CD40 protein, which was increased by ox-LDL. CONCLUSION: Protocatechualdehyde has protective effect on the CRL-1730 induced injury by ox-LDL and its mechanism of action may be related to the CD40/CD40L pathway. PMID- 18422187 TI - [Experimental research about the effect of Rehmannia glutinosa on the drug effect fingerprints of the rabbit's blood serum of Yin deficiency and heat flourishing syndrome]. AB - OBJECTIVE: To study about the effect of Rehmannia glutinosa Libosch on the drug effect fingerprints of the rabbit's blood serum of yin deficiency and heat flourishing syndrome. METHODS: The infusion of furosemide and escherichia coli endotoxin were used to replicate rabbit' s yin deficiency and heat flourishing syndrome animal model; high performance liquid chromatography gradient elution was adopped to study the change of the rabbit serum drug effect fingerprint in copying between before and after model-making and administration. RESULTS: Rehmannia glutinosa Libosch mainly affected metabolism level of the inner substance with the function of nourishing yin clearing heat. CONCLUSION: The serum HPLC fingerprint established is capable to reflect the function of nourishing yin clearing heat of Rehmannia glutinosa Libosch; the further study on its drug effect fingerprint will help to clarify its activity and metabolism of action. PMID- 18422188 TI - [Study on depressant effects of THPS on S180 tumor-bearing-mice and its mechanisms]. AB - OBJECTIVE: to study depressant effect of total hedysarum polybotys saccharids (THPS) on S180 tumor-bearing-mice and its mechanisms. METHODS: THPS was extracted from Radix Xedysari with water and precipitated with ethanol, determining its molecular weight, purity, saccharide and aldonic acid content. 90 Kunming mice were divided into 9 groups randomly. One group was the normal group, the others were divided into 8 groups randomly after inculating S180 tumors and were treated with THPS and THPS combination cyclophosphamide (CY) in low, moderate and high dose, to put them to death after 14 days. To determine every tumor weight, the rate of depressant tumor, the contents of IL-2 and TNF-alpha with ELISA, and NF kappaB with immunochemistry. RESULTS: The moderate dose THPS conspicuously possessed a depressant effect on S180 tumor and joint action with combination CY and increased the contents of IL-2, TNF-alpha and NF-kappaB of mice. CONCLUSION: THPS of moderate dose possesses a depressant effect on S180 tumor through increasing the contents of IL-2, TNF-alpha and NF-kappaB of mice. PMID- 18422189 TI - [The protective effect of Astragalus membranaceus extraction on cryopreserved primary-cultured human fetal hepatocytes]. AB - OBJECTIVE: To observe the influence of Astragalus membranaceus Extraction (AE) on the primary-cultured human fetal hepatocytes stored in liquid nitrogen and explore a new method for the cryopreservation of human hepatocytes with improved function. METHODS: Human fetal hepatocytes were harvested by two-step collagenase perfusion, and then stored in a liquid nitrogen for one month with five different cryoprotectants (I: 10% DMSO, II: 5% DMSO + 2 mg/L AE, III: 5% DMSO + 20 mg/L AE, IV: 5% DMSO + 60 mg/L AE, V: 5% DMSO + 100 mg/L AE). One month later, the cells were thawed rapidly and the viability, morphology and basic function of them were tested. RESULTS: The human fetal hepatocytes in different groups showed various levels of viability, morphological manifestation and cell function respectively. After thawing, the viability rate and flash adhering rate in group IV and V had no significant difference with group I (P > 0.05), but were higher than group II and III (P < 0.05); the cell function analysis in the group IV, the results of ALB and AST level determination, NH4Cl transformation test, were the best among the groups (P < 0.05). CONCLUSION: AE can provide protection for human fetal hepatocytes in cryopreservation, and the best performance concentration level of its is 60 mg/L; the preservation dosage of DMSO can be reduced when combined with AE in the preservation solution, which shows that AE has a synergistic effect with DMSO. PMID- 18422190 TI - [Effects of grape seed proanthocyanidins on oxidative stress in primary rat hippocampal neurons]. AB - OBJECTIVE: To investigate the effects of grape seed proanthocyanidins (GSP) on the oxidative stress induced by hydrogen peroxide (H2O2) in primary rat hippocampal neurons. METHODS: Primary cultured hippocampal neurons injury model by H2O2 was established. Morphological changes of neuron cells were visualized. The viability of hippocampal neurons was detected by MTT. The contents of malonidaldelyde (MDA), reactive oxygen species (ROS), superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-Px) in the cells were measured by chemochromatometry respectively. RESULTS: The cell activity decreased remarkably induced by 1 mmol/L H2O2 for 24 hours (P < 0.01), the levels of MDA and ROS were increased, and SOD, CAT, GSH-Px level were decreased in cells. GSP could reduce MDA and ROS level obviously, and increase SOD, CAT and GSH-Px level in cells. CONCLUSION: GSP has protective effect on hippocampal neurons injury induced by H2O2. The protective effect may be related to protecting cell membrane, increasing the activity of clearance enzyme of free radical and inhibiting lipid peroxidation. PMID- 18422191 TI - [Tissue distribution on methyl nonyl ketone of the volatile oil from Houttyunia cordata in mice]. AB - OBJECTIVE: To study the tissue distribution on methyl nonyl ketone of the volatile oil from Houttuynia cordata in mice in order to provide a guidance for the clinical trial. METHODS: The concentrations of the volatile oil from Houttuynia cordata in biological samples were determined by GC method. RESULTS: After a single oral dose of 5.0 g/kg volatile oil from Houttuynia cordata in mice, parent drug was mainly distributed in windpipe, intesting, liver, kidney, heart, blood, spleen, lung, brain, muscle; after 7 hours, parent drug of every tissue decreased over 90%. CONCLUSION: Parent drug mianly distributes in windpipe, intestine, liver, kidney. The experiment provides pharmacokinetic evidence for the rational administration and the further development of Houttuynia cordata. PMID- 18422192 TI - [Comparison of activities of various species of Rhizoma Cimicifugae and their honey processed products]. AB - OBJECTIVE: To compare the analgesic and sedative effects of various species of Rhizoma Cimicifugae and their processed products with honey. METHODS: The three kinds of analgesic experiments of formalin, hot plate and writhing tests, and the number of spontaneous activities and raise forelimbs were adopted in mice. RESULTS: All samples showed antinociceptive activities in the two phases of formalin, tail immersion and writhing tests and sedative properties in the spontaneous locomotor activity test. The analgesic and sedative effects of processed Rhizoma Cimicifugae were higher than crude drugs. CONCLUSION: The present results demonstrate that the antinociceptive and sedative activity of samples processed with honey are more potent than crude drugs. PMID- 18422193 TI - [Enrichment process of gentiopicroside from the leaves of Gentiana macropylla with macroporous resin]. AB - OBJECTIVE: To select the best type of macroporous resin for enriching gentiopicroside from the leaves of Gentiana macropylla, and study its optimal conditions and parameters. METHODS: Adsorption and desorption characteristics of gentiopieroside were carried out on the resins. HPLC was used to determine the content of gentiopieroside. RESULTS: The better resin D-101 was chosen, and the optimal parameters were obtained. CONCLUSION: The process is feasible to enrich gentiopicroside from the leaves of Gentiana macropylla Pall. PMID- 18422194 TI - [Ultrasonic extraction for flavonoids from Perilla frutescens by response surface method]. AB - OBJECTIVE: To investigate the extraction process of Perilla frutescens flavonoids. METHODS: Response surface method (RSM) was applied to a fractional design. The Perillafrutescens flavonoids were extracted by ultrasonic method. The influence factors of concentration of alcohol, ratio of liquid to solid, extraction times were evaluated. RESULTS: The optimum extraction conditions were confirmed as follows: 60% alcohol extracted 4 times, 18 minutes every time and ratio of liquid to solid was 15:1. The extraction rate of flavonoids under this condition was 24. 65 mg/g. CONCLUSION: Applying ultrasonic method to extract Perilla frutescens flavonoids is feasible. PMID- 18422195 TI - [Application of fingerprint of active part on extraction techniques for Vernonia anthelmintica]. AB - OBJECTIVE: To probe into feasibility of extraction techniques for Vernonia anthelmintica using HPLC fingerprint of active part as model. METHODS: The extraction techniques were studied by multi-index test experiment formula evaluation and orthogonal design, which took change of peak area in HPLC fingerprint of active part for Vernonia anthelmintica as indices. RESULTS: The optimum extracting procedure was as follows: using 10 times volume of water for each time, soaking one hour and then boiling for three times sustaining one hour each. CONCLUSION: It is scientific, simple and applicable. PMID- 18422196 TI - [Extracting flavonoids from Smilax glabra by microwave-assisted method]. AB - OBJECTIVE: To optimize the process of extracting flavonoids from Smilax glabra. METHODS: Flavonoids were extracted from Smilax glabra by microwave-assisted method, and the extracting time, microwave power, ethanol concentration, solid solvent ratio and extracting temperature were optimized through single factor experiment and orthogonal test. RESULTS: The optimun process parameters of extracting flavonoids were as follows: the extracting time, microwave power, ethanol concentration, solid-liquid ratio and extracting temperature were 5 minutes, 600 W, 60%, 1:30 and 80 degrees C, respectively. CONCLUSION: The proposed method has been applied stably and reliably to the extraction of flavonoids from Smilax glabra which has the advantages of high recovery and short extraction time. The recovery, the yield and the content of falvonoids are 93.2%, 2.66% and 32.6%, respectively. PMID- 18422197 TI - [Study on the formulation and process of Da Chuan Xiong Fang pellet by extrusion spheronization]. AB - OBJECTIVE: To study on the formulation and process of Da Chuan Xiong Fang pellet by extrusion-spheronization. METHODS: Based on the physical characters of active parts, the fast-release pellets of Chuan Xiong and Tian Ma were prepared separately by extrusion-spheronization. The formulation and process were studied by direct comparison. Then their characters were evaluated. RESULTS: The active components could release more than 90% in 30 minutes and the pellets presented perfect sphericity and narrow diameter distribution. CONCLUSION: Pellets of TCM can be prepared by homemade extrusion-spheronization machine and the fast-release pellets show perfect sphericity and narrow diameter distribution. PMID- 18422198 TI - [Preparation of total matrine controlled release micropellet capsules]. AB - OBJECTIVE: To prepare total matrine controlled release micropellet capsules. METHODS: The capsules were prepared by the technology of film-coating with surelease as matrix, and judged by the release behavior. RESULTS: The drug had completely been coated into a homogeneous and continuous film, and the results repeated in three batches showed that the release behavior of the pellet capsules followed zero-equation. CONCLUSION: The capsules have good sustained release effect and the process of the preparation is available. PMID- 18422200 TI - Influenza. PMID- 18422199 TI - [Observation on the expression of PS2 and platelet activating factor affected by Jianwei Yuyang granules and study its potential mechanisim against peptic ulcer]. AB - OBJECTIVE: To observe the influences of Jianwei Yuyang (JWYY) granules on expressions of PS2 and platelet activating factor (PAF), and analyze its potential mechanism. METHODS: 76 gastric ulcer patients were final diagnosed by gastroscope. They were randomly devided into 3 groups, including JWYY group (36 cases), Ranitidine group (36 cases) and Normal group (12 cases). Detection on Biospy Specimens of gastric mucosa to obseve the change of PAF, PS2 and the contents of aninohexose and phosphatide in ulcerated gastric mucosa for Immunohischemical and Western blotting. RESULTS: The contents of aninohexose, phosphatide of PU patients increased in JWYY group. There was significant difference between JWYY group and Ranitidine group (P < 0.01). The Immunohischemical and Western blotting methods showed that there was a linear correlation between the the contents of phosphatide, anino-hexose and the expression of PAF, PS2 in PU patients (P < 0.01). CONCLUSION: JWYY granule can prevent the occurrence and relaps of ulcer and affect the hydrophobicity of gastic mucosa and strengthen the stability of myxo-gellayer by reducing the expression of PAF, elevating the expression of PS2 and effecting the contents of phosphatide and aninohexose, that may be one of the mechanisms of JWYY to heal ulcer quickly. PMID- 18422201 TI - The effect of intra-neural local anaesthetic infusion on pain following major lower limb amputation. AB - BACKGROUND: Critical lower limb ischaemia has an annual incidence of 500-1000 per million and around a quarter will undergo a major lower limb amputation. Post operative pain and phantom pain are recognised complications. AIM: The aim of this study was to assess the role of postoperative intraneural infusion of local anaesthetic in patients under-going major lower limb amputation. METHODS: Between May 1998 and July 2001, following amputation patients either had standard post operative analgesia or had an intra-neural infusion of 0.5% bupivicaine via an infant feeding catheter connected to a standard syringe pump (Alaris). This was a retrospective review and clinical notes of these patients were analysed to assess their post operative course, analgesic requirements and to ascertain phantom limb pain/sensation rates. All the data collected was from standard drug charts, case note entries and out-patient letters. RESULTS: During this time period 64 patients had a major lower limb amputation (31 patients treated routinely and 33 patients had an intra-neural anaesthetic (INA) catheter placed). In the INA group median post-operative opioid analgesia requirement was 10mg versus 74 mg (p=0.0002, Mann-Whitney U) and post-operative prescription of amitriptyline for phantom pain was less common (4 patients versus 11 patients; p=0.0281, Mann Whitney U). There was no obvious increase in complications or significant adverse events. CONCLUSIONS: Postoperative intra-neural local anaesthetic infusion is a safe and effective technique. It reduces post-operative opioid analgesia requirement and seems to reduce phantom pain development. PMID- 18422202 TI - Let's get physical! A study of general practitioner's referral letters to general adult psychiatry--are physical examination and investigation results included? AB - BACKGROUND/AIM: The authors previously conducted a survey of psychiatrists' attitudes to physical examination and investigations of out-patients. The most common reason for not performing such investigations was the expectation that they had already been undertaken by the general practitioner (GP). We decided to test this theory. METHOD: A series of GP out-patient referral letters to general psychiatry was examined to establish whether findings from physical examination and investigations had been included. RESULTS: One hundred and three letters were examined. None of the letters contained information relating to a physical examination. Only one in twenty had information on investigations despite 4 out of 10 patients in the sample presenting to the GP with somatic symptoms. CONCLUSION: Details of physical examination and blood tests are not routinely included in referral letters to general psychiatry. This may lead to missed diagnoses of primary or secondary physical illness in psychiatric presentations. Unless it is clearly stated in the GP referral letter, it is unwise to assume that necessary investigations to exclude physical causes of presenting symptoms have been performed. Suggestions are made to improve communication between GPs and psychiatrists. PMID- 18422203 TI - Encouraging attendance at outpatient appointments: can we do more? AB - BACKGROUND AND AIMS: Non-attendance at new outpatient appointments is a challenge for health services. Patients with treatable morbidity may fail to receive care from which they would benefit, and scarce resources are wasted. Therefore, a literature review was undertaken to assess the effectiveness of appointment reminders as a means of increasing attendance and reducing 'Did not Attend' rates at new outpatient appointments. METHODS: Electronic databases were interrogated using a combination of MeSH terms and truncated keywords. In addition, the reference lists of papers were studied to identify additional relevant material. RESULTS: Telephone and postal reminders appear effective, particularly if received within a day of scheduled appointments. Postal reminders offering a reward for attendance, threatening sanctions for non-attendance, and providing information about the clinic appear more effective than standard reminders. No studies assessing the impact of measures including e-mail or text messaging were identified. Many studies were conducted in psychiatric settings outwith the United Kingdom raising issues of generalisability. In addition, no assessment of how initiatives impact on inequalities in service access were found. CONCLUSIONS: There is scope to increase attendance and reduce non-attendance at outpatient appointments but initiatives should be piloted in conjunction with robust evaluative frameworks. PMID- 18422204 TI - An atypical case of neuralgic amyotrophy with respiratory muscle weakness: case report and review of literature. AB - This report is of an atypical case of neuralgic amyotrophy with a Horner's syndrome, bilateral brachial plexus, lumbar plexus and phrenic nerve involvement. The diagnosis isconfirmed based on a classical history and examination findings with typical neurophysiological investigations for this condition. This report also highlights the novel use of positional magnetic resonance imaging to investigate patients with respiratory muscle weakness. This case report expands the recognised clinical features of neuralgicamyotrophy and the literature concerning atypical features of this condition is reviewed. PMID- 18422205 TI - Drug-eluting stents: a review of current evidence on clinical effectiveness and late complications. AB - Drug-eluting stent (DES) use has increased greatly as a result of early trial evidence of a reduction in restenosis. However, thet are expensive and do not improve patient survival. Therefore their use has been rationed in some countries. There is a paucity of clinical evidence for some patient groups such as non-ST elevation myocardial infarction and multi-vessel disease. Recent studies suggest that the early benefits of drug-eluting stents may be offset by an increased risk in late stent thrombosis which is a potentially fatal complication. However, the absolute risk appears low and, as yet, there is no evidence of an increased risk of stent-thrombosis related myocardial infarction or death in patients studied in randomised clinical trials. Long-term use of anti platelet therapy may protect against the risk of late stent thrombosis but the optimal treatment strategy is currently unclear. The aim of this paper is to provide an up-to-date review of the current evidence on DES; including clinical effectiveness, the limitations of existing trials, the emerging evidence on late stent thrombosis and the potential role of clopidogrel. PMID- 18422206 TI - The management of renal failure in patients with liver disease. Experiences from a district general hospital. AB - AIM: To compare the pre-existing management of patients with Hepatorenal Syndrome (HRS) in the gastroenterology unit of the Royal Alexandra Hospital, Renfrewshire, with the published evidence based studies. METHOD: A retrospective, 6-month, case record review of patients diagnosed with HRS was performed. An evidence-based protocol for the diagnosis and management of HRS was introduced into the unit, to aid patient treatment prospectively. After 6 months, both compliance with the protocol, and patient outcomes were analysed. RESULTS: Eleven patients were identified in the first part of the audit cycle, all of whom died. Seven were identified in the second cycle. Two had their renal function successfully corrected and one was discharged from hospital. Renal impairment and staging of liver disease was equivalent in both groups. The second group received more appropriate and aggressive therapy. Alcohol was the causative aetiology of liver disease in all patients. CONCLUSIONS: Targetted therapy in patients with severe liver disease and HRS can improve renal parameters. Previous studies have shown this to be linked with improved patient outcomes. PMID- 18422207 TI - Knowledge about genital herpes amongst first time attenders at a department of genitourinary medicine. AB - BACKGROUND AND AIMS: Genital herpes is the most common cause of genital ulceration in the developed world, and its prevalence in the United Kingdom is rising. Public knowledge of the infection is often limited. We aimed to assess knowledge among patients attending an outpatient genitourinary clinic in Scotland. METHODS: A questionnaire was designed to collect information about educational qualifications and knowledge of genital herpes. In the knowledge section, each correct answer was given a score of 1; thus the maximum possible score was 12. Two hundred and ten individuals who attended as new patients at a walk-in genitourinary medicine clinic were invited to complete the questionnaire. RESULTS: Two hundred and seven patients completed the questionnaire. Overall 63% of responses were answered correctly. Knowledge of genital herpes was better in patients who worked in healthcare (8.3 v 6.9, p=0.019) and in patients with educational qualifications gained from college or university compared to those gained at school (7.6 v 6.5, p=0.009). Level of knowledge was not related to age or gender; receiving sex education at school did not appear to improve knowledge of genital herpes. CONCLUSION: Knowledge of genital herpes among patients attending an outpatient genitourinary clinic in Scotland is reasonable but needs to improve to combat the rising prevalence of the infection. The areas of misunderstanding identified in our study could be targeted for public education. PMID- 18422208 TI - Laparoscopic hernia repair. AB - The laparoscope was first invented in the early 20th century. The first laparoscopic hernia repair was in 1990. Despite almost 20 years of laparoscopic hernia repair experience, open hernia surgery remains the main stay of hernia surgery today. This review looks at current evidence and guidelines on the use of laparoscopic hernia repair surgery. PMID- 18422209 TI - A review of surgical nutrition. AB - Malnutrition remains a common problem in surgical patients and is associated with significant morbidity and mortality. It is imperative that all surgical patients undergo nutritional screening on admission to highlight malnourished or at risk patients and implement a nutritional plan. Nutrition can be delivered by oral supplements, enteral or parenteral feeding, the route depending on an individual's requirements and surgical condition. Enteral feeding has largely been regarded as superior to parenteral feeding, as it is cheaper, safer and "more physiological" but studies show this is not always the case. This article reviews the basics of surgical nutrition and assesses the evidence supporting enteral versus parenteral nutrition. PMID- 18422210 TI - Dr John Rankin; his life, legacy and the 50th anniversary of the Rankin Stroke Scale. AB - Dr John Rankin (1923-1981) is one of the many distinguished alumni of the former University Department of Materia Medica and Therapeutics, Stobhill Hospital Glasgow. While his varied international career encompassed pulmonary physiology, occupational medicine and public health, he remains best remembered in the United Kingdom for his early stroke publications. In a series of articles published 50 years ago in the Scottish Medical Journal he described early rehabilitative stroke medicine using a novel grading system. Half a century on Rankin's eponymous stroke scale has become the endpoint of choice in acute stroke trials. This paper describes Rankin's remarkable career and the legacy of his work, with a particular focus on his stroke research and grading system. PMID- 18422211 TI - Coventina's column. PMID- 18422212 TI - Say no to 'Martini' health care. PMID- 18422213 TI - Management of HIV: still a long way to go. PMID- 18422214 TI - Sequential vs combination colorectal cancer therapy sparks debate. PMID- 18422215 TI - HIV and respiratory medicine. AB - The end of the 20th century saw the start of an HIV pandemic that brought death and suffering to millions. Anti-HIV drug therapy has improved the lives of many, although HIV-related respiratory complications remain extremely common worldwide. PMID- 18422217 TI - The genitourinary complications of HIV infection in men. AB - This article discusses infective and malignant complications of HIV affecting the genitourinary tract in men. Immunosuppression increases both the frequency of infections, and the range of organisms that may be involved. Cancers are common and presentations may be atypical. PMID- 18422216 TI - HIV and the kidney. AB - Kidney disease is an important complication of HIV infection. Antiretroviral therapy has dramatically improved the life expectancy of HIV-infected patients with end-stage renal disease. Renal replacement therapy, including kidney transplantation, should be offered to HIV-positive patients. PMID- 18422218 TI - Restless legs syndrome: symptoms, management and new developments. AB - Restless legs syndrome is a common but misdiagnosed disorder. Symptoms include an intense desire to move the legs when a patient is sitting or lying which is relieved by leg movement or walking. The condition is associated with abnormalities of the dopaminergic system. Drug treatment can be effective but should be confined to more severe cases. PMID- 18422219 TI - Facial nerve palsy: assessment and management. PMID- 18422220 TI - Gynaecological ultrasound. PMID- 18422221 TI - Time management: a guide for the foundation year doctor. PMID- 18422222 TI - Laboratory medicine: a guide for foundation doctors. PMID- 18422223 TI - Advice for junior doctors working in a trauma unit. PMID- 18422224 TI - Non-invasive imaging of coronary artery disease. AB - Non-invasive imaging of the heart is an expanding field in cardiology and appropriate test selection depends on the clinical question and test availability. This article reviews the role of single photon emission computed tomography myocardial perfusion scintigraphy, positron emission tomography and multi-detector computed tomography. PMID- 18422225 TI - Bisphosphonate-related osteochemonecrosis of the jaws. AB - Osteochemonecrosis of the jaws is now a well-known complication of bisphosphonate use. It can present management difficulties for doctors and dentists alike. AIl doctors prescribing bisphosphonates have a responsibility to patients to inform them of the risks of this complication no matter how small. PMID- 18422226 TI - Modernising medical careers and factors influencing career choices of medical students. AB - This article details medical students' views towards future career choices and factors that may influence this choice. The role of gender in career choice and the importance of structured career advice and management is highlighted and discussed. PMID- 18422227 TI - A simple case of heart failure? PMID- 18422228 TI - Bilateral ankle pain and quinolone use: a case of tendon rupture secondary to quinolone use. PMID- 18422229 TI - Elevated alkaline phosphatase: a diagnostic pearl for Paget's disease. PMID- 18422230 TI - Cerebral venous sinus thrombosis in polycythaemia. PMID- 18422231 TI - Completing surgical management of morbid obesity. PMID- 18422232 TI - Investments and risk. PMID- 18422233 TI - Francis Joseph Gall (1758-1828): false prophet of cerebral localization. PMID- 18422234 TI - Is cricoid pressure needed during rapid sequence induction? PMID- 18422235 TI - [Oral contraceptives give long-lasting protection against ovarian cancer. Use of oral contraceptives can explain reduced incidence of ovarian cancer in Sweden]. PMID- 18422236 TI - [Long-lasting pain in childhood and adolescence can and should be treated. The problem is common and leads to great suffering]. PMID- 18422237 TI - [Few pain clinics for children. Specialist competence needed at all pediatric clinics in the country]. PMID- 18422238 TI - [Increasing workload in psychiatric emergency services. More visits, but the number of first time visitors is decreasing]. PMID- 18422239 TI - [Current Swedish renal function tests must be better. Uncertain diagnosis with routine formulas--great risk of wrong drug dosages]. PMID- 18422240 TI - [Discussed--but mini-incision hip arthroplasty here to stay]. PMID- 18422241 TI - [The Medical Products Agency is responsible for the control of medical technological products: we need more active medical professionals reporting shortages]. PMID- 18422242 TI - [Paternalism and prenatal diagnosis]. PMID- 18422243 TI - [Will private practitioners be allowed to have a look at proposal for a new national reimbursement system?]. PMID- 18422244 TI - [There are limits to altruism--clinical research must be profitable]. PMID- 18422245 TI - [Annual continuing education for specialists]. PMID- 18422246 TI - [The patent is "engine" in drug development]. PMID- 18422247 TI - [Medical angiology education for all specialists in internal medicine]. PMID- 18422248 TI - [Unfortunate if the interpretation of HSAN's decision should be that subclinical hypothyroidism always be treated]. PMID- 18422249 TI - [Billroth and Brahms. A problematic friendship between two icons]. PMID- 18422250 TI - Development and survival of Haemonchus contortus infective larvae derived from sheep faeces under sub-tropical conditions in the Potohar region of Pakistan. AB - Assessment on the development and survival of Haemonchus contortus larvae was made to evaluate the influence of microclimatic variables viz., relative humidity (%), temperature (degreesC), rainfall (mm) and intensity of sunlight (hrs). Pasture plots in a subtropical area, Pakistan were contaminated with H. contortus eggs at the start of every month. The plots were sampled on fortnightly basis after contamination for three consecutive months. The overall pattern indicated the most favorable conditions for survival, development and translation to herbage was during the wet season from July to October and the least favorable was during the dry season from April to June. Peak infective larvae (L3) recovery was during the 15-45 day interval for each plot and the lowest number was during the 75-90 day interval. Herbage was collected in the morning, afternoon and evening and greatest recovery of L3 was in the morning time and least in the afternoon. The number of L3 on pasture was directly related to the pattern of rainfall and relative humidity with a significant (P<0.05) positive correlation and temperature and intensity of sunshine were negatively correlated (P<0.05). The results suggest that pastures can remain infective for up to 2 months and become relatively clean by 90 days after contamination. Thus, long pasture rest periods, especially during the high risk wet season, may be a helpful strategy to reduce the infection levels. PMID- 18422251 TI - Acute coliform mastitis in buffaloes (Bubalus bubalis): clinical findings and treatment outcomes. AB - This report was delineated to study the clinical, bacteriological and therapeutic aspects concerned with acute coliform mastitis in buffaloes. Bacteriological examination of 80 quarter milk samples obtained aseptically from 56 buffaloes with acute mastitis revealed that coliform bacteria was the most common pathogen (45 cases) followed by Staphylococcus aureus (seven cases) then Streptococcus uberis (three cases), and Streptococcus agalactiae (one case). Clinically, hotness, swelling and painful reaction with serous excretion containing clots was recorded in buffaloes with coliform mastitis. The efficacy of ceftiofur was evaluated in the treatment of buffaloes with acute coliform mastitis. Parenteral ceftiofur neither improved clinical signs nor returned milk to pre-infection production level, whereas intramammary ceftiofur and combination of intramammary with parenteral ceftiofur improved the clinical signs in 10/15 and 12/15 buffaloes, respectively. On quarter level, 3/17, 12/17 and 15/21 quarters recovered in groups received parenteral, intramammary and combination therapy, respectively. This study demonstrates that systemic ceftofur is not effective in the treatment of clinical coliform mastitis in buffaloes. PMID- 18422252 TI - Prevalence of parasites of the local scavenging chickens in a selected semi-arid zone of Eastern Kenya. AB - A study to identify and estimate the prevalence of parasites of local chickens in a semi arid area of Kenya was conducted between March 2005 and August 2006. Three hundred and sixty (360) local chickens purchased from Yathui division of Machakos were examined. Of those, 93.3% had helminths. Nematodes were recovered in 268 (74.4%) chickens whereas 245 (68.1%) had cestodes. Tetrameres americana (37.7%), Ascaridia galli (33.3%) and Heterakis gallinarum (22.8%) were the most important nematode species identified. Raillietina echinobothrida (33.3%) and Davainea proglottina (19.4%) were the two most important cestode species identified. Two coccidia species, namely Eimeria necatrix (6.7%) and E. tenella (16.7%) were isolated and identified as per location in the digestive system. Important ectoparasites identified included Echidnophaga gallinacea (76.7%), Menacanthus stramineus (79.4%) and Dermanyssus gallinae (60.0%). Endo-parasites (helminths and coccidia) occurred in significantly (p<0.05) higher frequencies during the wet season than during the dry season. On the contrary, ecto-parasites were significantly (p<0.05) more fequent during the dry season. Male chickens generally exhibited increased odds for the occurrence of parasites than female birds. Further investigations are required to establish a plausible explanation for this. Overall, parasitism was a big constraint to chicken productivity in the study area. Urgent integrated parasite control approaches should be initiated to address parasitism in chickens in the Yathui cluster. PMID- 18422253 TI - Ameliorative effects of boron on serum profile in buffalo (Bubalus bubalis) fed high fluoride ration. AB - An experiment was undertaken to evaluate the protective role of boron on the serum profile of buffalo calves fed a high fluoride ration. Twelve male Murrah buffalo (Bubalus bubalis) calves of 6-8 months age, divided into three groups of four calves in each, were fed basal diets and supplemented with sodium fluoride (NaF, 60 ppm) alone or in combination with borax (Na2B4O7.10H2O, 140 ppm) for 90 days. Boron (B) was added in the ration as borax to make @140 ppm boron (elemental B) on DM basis in treatment II. Dietary F caused a significant (p<0.05) depressing effect on serum Ca and Zn on day 90 which was improved with B supplementation. However, serum Fe and Cu did not show any significant change on F or F+B supplementation. The serum ALP and phosphorus level were increased significantly (p<0.05) on F feeding but declined significantly (p<0.05) when B was fed. The findings suggested beneficial effect of boron on serum minerals and ALP in buffalo calves fed high fluoride ration. PMID- 18422254 TI - Sweet potato and Green panic as sole fodder for stall-fed lactating White Fulani cows and growing calves. AB - Three sweet potato varieties (TIS-87/ 0087; TIS-8164; TIS-2532.OP.1.13) and Green panic (Panicum maximum) were evaluated for forage quality in a completely randomized design (CRD) using 12 sole-fed lactating White Fulani cows for 21 days, and 12 growing White Fulani and White Fulani x Brown Swiss calves for 77 days. Dry matter intake of cows ranged (p<0.05) between 131 g/Wkg(0.734) in TIS 87/0087 and 152 g/Wkg(0.734) in TIS-8164. Daily milk yields increased (p<0.05) in TIS-87/0087 (2%) and TIS-8164 (5%), but decreased in TIS-2532.OP.1.13 (5%) and Green panic (19%). Mean (+/-SEM) milk total solids (13.16+/-0.05 g/100 g), ash (0.78+/-0.01 g/100 g), protein (3.79+/-0.07 g/100 g), fat (3.98+/-0.06 g/100 g), and lactose (4.62+/-0.14 g/100 g) were significantly different (p<0.05) among treatments. The White Fulani x Brown Swiss calves recorded about 10, 25 and 29% higher birthweight, milk intake and bodyweight gains respectively, but about 3% lower dry matter intake than the purebred White Fulani calves. The rumen degradation rates were similar (p>0.05), while the rest rumen dry matter degradation characteristics were lowest (p< 0.05) for Green panic. Sweet potato forage could be used as sole feed or supplement for dry-season Green panic. PMID- 18422255 TI - Herd dynamics and contribution of indigenous pigs to the livelihoods of rural farmers in a semi-arid area of Zimbabwe. AB - Herd dynamics of indigenous pigs were determined in a semi-arid farming area of Chirumanzu, Zimbabwe. Thirty-two pig herds were monitored once every month for 12 months. Consequently, pig production potential (PPP) and pig production efficiency (PPE) were computed for each month. Incidences of farrowing were higher in times of pig confinement (57%) than in times when the pigs were free ranging (43%). About 23% of the farmers interviewed had access to irrigation. The mean pig herd size per household was 3.3. The average litter size per farrowing was 7.7. Pigs were free ranging for most of the year and penned during the rainy season. Herd sizes changed with month (P<0.05). Farmers who had access to irrigation had higher (P<0.05) herd sizes, number of piglets and the number of adult pigs. Highest piglet numbers were observed during the rainy months. The number of mature pigs contributing to pig production efficiency was low. Piglet mortality was the major cause of exits from the herd and was higher in farmers with no access to irrigation facility (P<0.05). The PPP was highest in farmers who had access to irrigation (P<0.05), except in June and July. The PPE was low and not affected by either month, village or access to irrigation (P>0.05). It can be concluded that pig production efficiencies were low and intervention strategies to improve them need to be developed. PMID- 18422256 TI - Seroprevalence of egg drop syndrome--76 virus as cause of poor egg productivity of poultry in Nsukka, South East Nigeria. AB - To determine if egg drop syndrome 76 virus infection is among the causes of lowered egg productivity in commercial poultry farms in South Eastern Part of Nigeria and to know the prevalence of the infection, ten farms with history of lowered egg production in Nsukka local government area of Enugu State were randomly selected. Sera from ten hens in each of the selected farms were assayed for antibodies against EDS 76 virus by the haemagglutination-inhibition (HI) test. The mean HI titre of the ten hens in each of the farms was recorded as EDS 76 antibody titre for the farm. Nine out of the 10 farms tested were positive for EDS - 76 antibodies with HI titres ranging between 16 and 256. Out of 10 flocks with production of 65% and above 9 were EDS-76 HI negative. PMID- 18422258 TI - Factors affecting milk yield, milk fat, bacterial score, and bulk tank somatic cell count of dairy farms in the central region of Thailand. AB - A study was conducted to determine the effects of season, farm location, and farm size on farm milk yield (FMY), average milk yield per cow (AYC), milk fat, bacterial score, and bulk tank somatic cell count (BTSCC) of dairy farms in the central region of Thailand. Farms were located in the districts of Kaeng Khoi, Muaklek, Pak Chong, and Wang Muang. Collection of data was at the farm level; individual animal records were unavailable. A total of 967,110 daily farm milk yield, 58,575 milk fat and bacterial score, and 24,109 BTSCC records from 1,034 farms were collected from July of 2003 to June of 2006. There were three seasons: rainy, summer and winter. Farms were categorized into small, medium, and large according to the number of cows milked per day. Results showed that FMY and AYC were higher (p<0.05) in winter and lower in the summer and rainy seasons. In addition, the majority of small size farms had higher (p<0.05) AYC and milk fat values, and lower bacterial score and BTSCC values than medium and large size farms. PMID- 18422257 TI - Serum igG, blood profiles, growth and survival in goat kids supplemented with artificial colostrum on the first day of life. AB - The objective of this study was to compare serum IgG concentrations, blood metabolites indicative of nutritional status, weight gain and mortality rate in goat kids fed a commercial colostral supplement containing immunoglobulins against several pathogen microorganisms, prior to the ingestion of the mother colostrum, and goat kids ingesting natural colostrum only. There was no difference in serum IgG concentrations between 27 kids fed a colostrum supplement (20 g, derived from cow lacteal secretions) prior to the kids' first meal (658+/ 703 mg dl(-1)) and 21 kids ingesting maternal colostrum freely (1011+/-1140 mg dl(-1)) at 24 hours of birth. Hematocrit values, serum glucose and urea concentrations at 24 hours and 5 days of age were unaffected by treatment. Serum total proteins were 14% higher (P<0.05) in the unsuplemented group than in the supplemented group at 5 d of age. There was no significant difference between the supplemented and unsupplemented kids in daily weight gain from birth to 70 days of age (92+/-4.8 vs 102+/-5.1 g day(-1)). Mortality was 4% for kids receiving the colostrum supplement as compared with 0.0% for kids ingesting maternal colostrum only. Results suggest that, in intensively managed non-dairy goats with kiddings in summer, the supplementation of this commercial colostrum derived from cow lacteal secretions and containing antibodies against diverse pathogens organisms did not enhanced growth, survival or immunity under the farming conditions of this study. PMID- 18422259 TI - Intake and digestibility in sheep and chemical composition during different seasons of some West African browse species. AB - Foliage of Afzelia africana, Pterocarpus erinaceus and Khaya senegalensis, from 10 trees per species, was collected every two weeks during the late dry, rainy and cool season to determine the seasonal effects on chemical composition. Fifteen rams of the Djallonke breed, weighing on average 20.0 kg, were used to evaluate the voluntary intake and digestibility of hay of A. gayanus, foliage of A. africana (as a sole feed), and A. africana, P. erinaceus and K. senegalensis offered with 30% of the diet as A. gayanus hay. The crude protein (CP) content ofA. africana, and P. erinaceus decreased significantly from the late dry season to the cool season when that of K. senegalensis tended to increase. The mean CP of A. africana, P. erinaceus and K. senegalensis differed significantly (173 g, 139 g and 114 g/kg DM, respectively). The DM intake ofA. africana offered with hay (571 g/d) or as a sole feed (598 g/d) were not significantly different, but was higher than that of P. erinaceus (428 g/d) and K. senegalensis (298 g/d). The digestibility calculated by difference of DM and CP of A. africana (582 g/kg DM and 795 g/kg CP, respectively) did not differ significantly from A. africana as a sole feed, but were higher than for the other species. The nutritive value of A. africana seems to justify the high preference of herders for this species. PMID- 18422260 TI - E-nirvana: are we there yet? PMID- 18422261 TI - Do we need Medicare? PMID- 18422262 TI - Discussing end-of-life care with your patients. PMID- 18422263 TI - Stark III: refinement not revolution (part 1). PMID- 18422264 TI - Beyond EHRs: how technology can help you treat chronic illness. PMID- 18422265 TI - Have you really addressed your patient's concerns? PMID- 18422266 TI - Looking out for others. PMID- 18422267 TI - [Enhancing study on drug metabolism in pharmaceutical sciences]. PMID- 18422268 TI - [Alternatively spliced variants of constitutive androstane receptor in liver of mouse]. AB - OBJECTIVE: To investigate the existence of alternatively spliced variants of constitutive androstane receptor (CAR) in liver of mouse. METHODS: The nucleotide from liver of mouse was purified and the CAR cDNA was amplified by PCR. The fragments of CAR cDNA were cloned to T vector and sequence analysis was performed. RESULT: Various spliced variants of CAR in liver mouse were confirmed by DNA sequencing. CONCLUSION: There are alternatively spliced variants in CAR, which are located in the ligand binding sequence of CAR. PMID- 18422269 TI - [Expression of human CYP2E1 in insect cells using bac-to-bac expression system]. AB - OBJECTIVE: To obtain recombinant human CYP2E1 and to determine its activity by using the specific probe substrate. METHODS: CYP2E1 cDNA was obtained by RT-PCR using human liver RNA as template. The cloned CYP2E1 cDNA was ligated with pFastBac vector to generate recombinant pFastBac-CYP2E1, which was then transformed into E. coli DH 10 Bac. Recombinant Bacmid-CYP2E1 was generated by transposition. Then Spodoptera frugiperda (Sf9) insect cells was infected with Bacmid-CYP2E1 to generate recombinant baculoviruses carrying human CYP2E1 cDNA. Finally, Sf9 insect cells were triinfected with recombinant baculoviruses carrying human CYP2E1, CYPOR and CYPb5. The activity of the recombinant enzymes was determined using chlorzoxazone as the substrate. RESULT: The Kmand Vmaxof recombinant CYP2E1 to chlorzoxazone was (72.4 +/-8.7) micromol. L(-1) and (2.41 +/-0.10) micromol.min(-1)?g(-1)protein, respectively. CONCLUSION: Active recombinant CYP2E1 has been obtained by bac-to-bac expression system and its activity is similar to previous reports. PMID- 18422270 TI - [Construction, expression and characterization of recombinant fusion protein HSA PTH (1-34) in Pichia pastoris]. AB - OBJECTIVE: To obtain recombinant fusion protein HSA (human serum albumin)-PTH(1 34) in Pichia pastoris. METHODS: HSA and PTH(1-34) cDNA were obtained with PCR and the DNA segments were cloned into vector pPIC9 with linker. The linearized plasmids were transformed GS115 competent cells treated with LiCl, and mut+ transformants were screened on MD plate. With AOX promoter and alpha-MF signal sequences leading, fusion protein was expressed in GS115. PCR and SDS-PAGE were employed to confirm the integration and expression of HSA-PTH(1-34). The fusion protein was identified by Western blotting and classical adenylate cyclase assay. RESULT: The PCR results showed that the gene of HSA-PTH(1-34) was integrated into GS115 genome. Western bolt approved the existence of two domains of HSA and PTH(1 34). The bioactivity assay in rabbit cortical membranes indicated that HSA-PTH (1 34) activated adenylate cyclase, but the activity was lower than that of the synthetic PTH(1-34). CONCLUSION: Active fusion protein HSA-PTH (1-34) is successfully expressed in Pichia pastoris. PMID- 18422271 TI - [Inhomogeneous expression of fusion protein HSA/IL1ra in Pichia pastoris]. AB - OBJECTIVE: To investigate the causes and influencing factors of heterogeneity of HSA/IL1ra fusion protein expression in Pichia pastoris. METHODS: The heterogeneity of HSA/IL1ra fusion protein expressed in Pichia pastoris was studied by removing glycosylation and inhibiting glycosylation, as well as by different ways of fusion, different clones, and different expression host. RESULT: Glycosylation caused expression heterogeneity of fusion protein, but in SMD1168 and some GS115 clones there was no this phenomenon. CONCLUSION: The expression heterogeneity of HSA/IL1ra fusion protein in Pichia pastoris is due to the glycosylation, and different ways of fusion, different clones, different expression host also have some impact. PMID- 18422272 TI - [Luteolin excretion after oral administration of Elsholtzia blanda benth extracts in rats]. AB - OBJECTIVE: To observe the excretion of luteolin after oral administration of Elsholtzia blanda benth extracts in rats. METHODS: Samples of urine, feces and bile were collected after oral administration of Elsholtzia blanda benth extracts in rate. After deconjugation with beta-glucuronidase/sulfatase, the levels of luteolin in urine, feces and bile were measured by RP-HPLC. RESULT: The recovery rate of luteolin was 98.0 %-106.0 % and the extract recoveries were 85.0 % 108.0%. Relative standard deviation (RSD) of intra-and inter-day assay was less than 10.0 %. The total accumulative excretion was 37 % (11 % in urine, 26 % in feces and bile). CONCLUSION: The established RP-HPLC method is sensitive, specific, accurate, and is applicable for determination of luteolin in rat urine,feces and bile. PMID- 18422273 TI - [Determination of progesterone concentration in plasma after oral administration of progesterone-loaded lipid nanoparticles in rats]. AB - OBJECTIVE: To establish a RP-HPLC method for determination of plasma progesterone and to apply the method for pharmacokinetics study of progesterone-loaded lipid nanoparticles after oral administration in rats. METHODS: The plasma samples were collected from castrated rat after oral administration of progesterone-loaded lipid nanoparticles and extracted by acetic ether. The determination was performed on a Hypersil C18 column (150 mm X 3.9 mm , 5 microm) with a mobile phase consisting of methanol and water (60:40) at a flow-rate of 0.6 ml/min. The UV detector was at 240 nm and danazol was used as internal standard. RESULT: Good linearity was obtained over the range of 0.02-2 microg/ml progesterone in plasma(r=0.9999, n=3). The quantification limit was (0.02 +/-0.004) microg/ml(n=3) and the limit of detection was 0.005 microg.mL(-1)(S/N = or >3). The inter-and intra-day RSDs were all less than 10% for quality control samples at high-, medium- and low-concentrations. The average absolute recovery rate was 90.5 % and the average method recovery was in the range of 93.4 %-107.5%. The plasma concentration-time curves indicated that tmax was delayed after administration of progesterone-loaded lipid nanoparticles, and the bioavailability was increased significantly, compared with contrast solution. CONCLUSION: The method developed is stable, simple, rapid, accurate, sensitive and applicable for determining plasma concentrations of progesterone of progesterone-loaded lipid nanoparticles in pharmacokinetic studies. PMID- 18422274 TI - [Metabolism-based interaction of diphenytriazol and flavone compounds]. AB - OBJECTIVE: To observe the metabolism-based interaction of diphenytriazol and flavone compounds. METHODS: Flavone compounds kaempferol, isoharmnten and Elsholtzia blanda benth extract were chosen as the substrate of glucuronidation in the phase II metabolism. The metabolism was investigated in different rat liver microsome incubates pretreated with beta-naphthoflavone (BNF), diphenytriazol and tea oil (control). The concentrations of residual substrate were determined by HPLC. Quercetin and kaempferol were coincubated with diphenytriazol in control microsome to evaluate the inhibition for phase I metabolism. The concentration of diphenytriazol was determined by HPLC. RESULT: The phase II metabolic activity of kaempferol, isoharmnten and Elsholtzia blanda benth extract in diphenytriazol-treated microsome was more potent than that in BNF-treated microsome (P<0.01). The phase I metabolism of diphenytriazol was markedly inhibited by quercetin and kaempferol, with the inhibition constants (Ki) (12.41 +/-0.26)microg . ml(-1) and (7.97 +/-0.08)microg . ml(-1), respectively. CONCLUSION: Diphenytriazol demonstrates metabolism-based interaction with flavone compounds in vitro. PMID- 18422275 TI - [Determination of dimethyl sulphate residual in granisetron hydrochloride by headspace gas chromatography]. AB - OBJECTIVE: To develop a headspace gas chromatography method for determining dimethyl sulphate residual in granisetron hydrochloride. METHODS: An Angilent INNOWAX capillary column with nitrogen gas as carrier and FID as detector was applied in this study. Dimethyl sulphate was tested under a constant column temperature. RESULT: Dimethyl sulphate had different retention time from other organic solvents such as alcohol,acetoacetate, isopropanol, dichlormethane and chloroform, which might exist in granisetron hydrochloride. The detection limit of dimethyl sulphate;s was 0.0016%. CONCLUSION: The method can be used for the determination of dimethyl sulphate residual in granisetron hydrochloride. PMID- 18422276 TI - [Determination of bis (p-fluorobenzyl) trisulfide and bis (p-fluoro-benzyl) disulfide in lungs of rat by HPLC]. AB - OBJECTIVE: To establish an HPLC method for analysis of bis(p-fluorobenzyl) trisulfide(BFTS) and bis(p-fluorobenzyl)disulfide(BFDS) in the lungs of rat. METHODS: 5.0 ml extract solvent (n-hexane: isopropyl alcohol=95:5, v/v) and 20 microl of 11.50 microg/ml dibenzyl disulfide (internal standard) were added to 0.2 g lung sample followed by homogenization. After centrifugation, 4.0 ml of supernatant was separated and vaporized to dryness, and the residue was reconstituted in mobile phase for HPLC analysis. The HPLC analysis was performed on an SB C18 column using acetonitrile and water (65:35, v/v) as mobile phase with a flow rate of 1.0 ml/min with UV detection at 220 nm. RESULT: The calibration curves for BFTS and BFDS in sample were linear over the concentration ranges of 0.04712-14.78 microg/g(r=0.999) and 0.04831-23.96 microg/g(r=0.999), respectively. The limits of quantification were 0.04712 microg/g and 0.04831 microg/g for BFTS and BFDS, respectively. The assay recoveries for BFTS and BFDS ranged from 95.71%-107.2% and 90.00%-110.5%, respectively. The precisions were obtained with RSD of <10%. The developed method was successfully applied to study the content of BFTS and BFDS in the lungs of rats after intravenous injection of 12.5 mg/kg BFTS. CONCLUSION: The method developed is simple, selective, repeatable and accurate, which can be applied to study the tissue distribution of BFTS and BFDS. PMID- 18422277 TI - [Differential expression of genes related to transcription in cultured hepatoma cells with intact genome of hepatitis C virus]. AB - OBJECTIVE: To investigate the effects of hepatitis C virus (HCV) on transcription regulation genes of host cells by gene chip assays in cultured cells with intact HCV genome. METHODS: Huh-7 hepatoma cells were cultured and infected with in vitro constructed HCV. The total RNAs, proteins and cell culture supernatants of HCV infected cells and control cells were isolated. Proteins and cell culture supernatants were used to detect the HCV replication and protein expression in cell culture system. The HCV protein expression was detected with Western blotting. Released HCV from infected cells was analyzed by real-time fluorescence quantitative PCR. Total RNA was qualified using 10 g/L agarose gel electrophoresis. cRNA was synthesized, fluorescence labeled and purified, then hybridized with Agilent oligo microarray (20173 probes). Differential expression of genes related to transcription in cell culture system was analyzed. RESULT: HCV was positive in cell culture supernatants and HCV protein expression was also positive according to Western blotting results. Eleven up-regulated and 11 down regulated genes related to transcription were found after Agilent gene chip screening. CONCLUSION: Intact hepatitis C virus cell culture system provides an useful tool for study on the affects of HCV infection on transcription regulation genes in host cells. PMID- 18422278 TI - [Expression of telomere binding factor 2 (TRF2) on leukemia cell lines and primary leukemia cells]. AB - OBJECTIVE: To detect the expression levels of telomere binding factor 2 (TRF2) on leukemia cell lines and primary leukemia cells. METHODS: The expression of TRF2 mRNA was detected with quantitative real-time RT-PCR in leukemia cell lines and primary leukemia cells. The Western blot analysis was used for the detection of TRF2 protein expression. RESULT: TRF2 was overexpressed in T-cell leukemia cell lines but not in myelogenous leukemia cell lines. Significant higher expression levels of TRF2 were observed in primary leukemia cells from patients with M0 and M1 subtypes of acute myelogenous leukemia (AML) compared with normal control and other subtypes of AML. CONCLUSION: Increased TRF2 expression levels are found in T-cell leukemia cell lines and AML patients with poor prognosis, which suggests that TRF2 expression might be related to the prognosis of leukemia. PMID- 18422279 TI - [Expression of glucose transporter in non-small cell lung carcinoma and its clinical significance]. AB - OBJECTIVE: To investigate the expression of glucose transporter (Glut)1, Glut3, and hypoxia inducible factor (HIF)-1 alpha in human non-small cell lung carcinoma (NSCLC), and its clinical significance. METHODS: Specimens of cancer tissues and paracancerous lung tissues from 34 cases of NSCLC and 17 specimens of benign lung lesions were collected. The expressions of Glut1, Glut3, and HIF-1 alpha were detected with immunohistochemical staining, RT-PCR, and Western blot. RESULT: The relative mRNA expressions of Glut1 and HIF-1 alpha were 0.689 +/-0.245, 0.693 +/ 0.248 in cancer tissues; and 0.338 +/-0.157, 0.351 +/-0.184 in paracancerous lung tissues (P <0.001); while those of Glut3 were 0.506 +/-0.246 in cancer tissues and 0.482 +/-0.238 in paracancerous tissues (P >0.05). The relative protein expressions of Glut1 and HIF-1 alpha were 0.582 +/-0.247, 0.525 +/-0.246 in cancer tissues and 0.288 +/-0.151, 0.261 +/-0.135 in paracancerous lung tissues (P<0.001), but the protein expressions of Glut3 were 0.551 +/-0.251 and 0.436 +/ 0.224 respectively (P>0.05). Glut1 and HIF-1 alpha expressions were higher in poor differentiation group and in stage III group, than those in medium and well differentiation group and stage I and II group. Moreover, there was a significant correlation between the expression of Glut1 and HIF-1 alpha (r=0.854, P<0.01). CONCLUSION: Glut1 and HIF-1 alpha are highly expressed in NSCLC, and their expressions are associated with tumor differentiation and clinical stage. PMID- 18422280 TI - [Effect of Skp2 antisense oligodeoxynucleotide on growth and proliferation of gastric carcinoma SGC-7901 cells]. AB - OBJECTIVE: To investigate the effect of S-phase kinase-associated protein 2 antisense oligodeoxynucleotide (Skp2 ASODN) on the growth and proliferation of gastric carcinoma SGC-7901 cells and its mechanism. METHODS: The Skp2 oligodeoxynucleotides (ODNs) were embedded in cationic liposome Lipofectamine 2000 reagent and transfected into SGC-7901 cells. The cell growth and proliferation were observed with light microscopy and MTT assay. Cell cycle was measured by flow cytometry. The expression levels of Skp2 and p27 mRNA were detected by reverse transcription-polymerase chain reaction. The expression levels of Skp2 protein and its substrate p27 protein were detected by Western blot. RESULT: After treatment with Skp2 ASODN, the growth and proliferation of SGC-7901 cells were inhibited in a dose-dependent manner with a peak value at 48 h. The inhibition rate of 200 nmol/L group at 48 h was 42.4 % (P<0.01). In cell cycle study the percentage of S phase cells in 200 nmol/L group was significantly higher than that in normal control group (P<0.05). Both Skp2 mRNA and its protein levels in 200 nmol/L group were significantly lower than those in control group and in Skp2 nonsense oligodeoxynucleotide (Skp2 NSODN) group (P<0.05). However, p27 mRNA level remained unchanged although its protein level was significantly higher than that in control group and NSODN group (P<0.05). CONCLUSION: Skp2 ASODN can inhibit the growth and proliferation of SGC-7901 cells, which may be mediated by interfering with ubiquitin-proteosome pathway and cell cycle regulation. PMID- 18422281 TI - [Effect of vitamin C and E on antioxidative enzyme, NOS activity and NO contents in hippocampus of rats with lead poisoning]. AB - OBJECTIVE: To observe the effect of vitamin C and E on blood lead (Pb) levels and SOD, GSH-Px, NOS activity and NO, MDA content in hippocampus of rats with lead poisoning. METHODS: Rat lead poisoning model was established by oral administration of 0.615 mmol/L lead acetate in drinking water for 4 weeks; and animals were fed with vitamin C 100 mg/kg. bw and/or vitamin E 100 mg/kg. bw for 1 week. Then blood Pb levels and SOD, GSH-Px, NOS activity and MDA, NO contents in hippocampus of rats were determined by corresponding kits. RESULT: Compared with control group, blood Pb level was decreased significantly (P<0.05) after given vitamin C, vitamin E or combination of vitamin C and E. The concentrations of SOD, GSH-Px, NO and NOS were significantly higher in vitamin C and/or E groups than those in control group (P<0.05). The concentration of MDA in vitamin treatment groups was significantly lower than that in lead control group (P<0.05); furthermore concentration of MDA in combination of vitamin C and E group was significantly higher than that in vitamin C alone group (P<0.05). CONCLUSION: Administration of vitamin C and E can decrease blood lead level, alleviate damage of lipid peroxidation in hippocampus by lead toxicity and reverse NO, NOS levels in rats with lead poisoning. PMID- 18422282 TI - [Effect of pyrrolidine dithiocarbamate on antioxidation of canine myocardium during ichemia/reperfusion injury]. AB - OBJECTIVE: To assess the alterations in myocardial energy metabolism and lipid peroxidation during canine cardiopulmonary bypass (CPB), and to investigate the interventional effects of pyrrolidine dithiocarbamate (PDTC) pretreatment. METHODS: Twelve adult healthy dogs undergoing CPB were randomized into control group (Group C, n=6) and PDTC group(Group P, n=6). In Group P, 30 mg/kg PDTC was administered intravenously before CPB and in Group C animals were given physiological saline instead of PDTC. The contents of adenosine triphosphate (ATP), superoxide dismutase (SOD), glutathione peroxidase (GSH-PX), malondialdehyde (MDA) and mitochondrial swelling degree (MSD) of myocardium were determined before CPB, 60 min after aortic cross-clamping (AC) and 60 min after declamping (DC). Hemodynamics was monitored before CPB, 30 min and 60 min after DC. RESULT: Contents of ATP, SOD and GSH-PX in Group P at 60 min after AC and 60 min after DC were higher than those in Group C (P<0.01). MDA and MSD in Group P at 60 min after AC and 60 min after DC were significantly lower than those in Group C (P<0.01). Hemodynamics of Group P was recovered at 30 min and 60 min after DC. CONCLUSION: Pretreatment with PDTC is effective in improving antioxidation capacity of myocardium and ameliorates myocardial energy metabolism. PMID- 18422283 TI - [Application of parasymphyseal mandible osteotomy combined with set-back tongue flap in treatment of tongue base carcinoma]. AB - OBJECTIVE: To evaluate the efficacy of the approach of parasymphyseal mandible osteotomy combined with set-back tongue flap in treatment of tongue base carcinoma. METHODS: In total 18 patients with cancer of tongue base, 13 were males and 5 females aged of 30 to 65 y (average 52.9 y). Eighteen patients were treated by parasymphyseal mandible osteotomy combined with set-back tongue flap in our hospital from June 2001 to June 2006. RESULT: The range of follow-up period was 2 to 5 years. All patients had satisfactory speech, swallowing and taste function. CONCLUSION: Parasymphyseal mandible osteotomy provides good exposure, and set-back tongue flap to reconstruction of the base of tongue can partially restore the function of tongue. PMID- 18422284 TI - [Value of 3D-DCE MRA in diagnosis of cavernous transformation of portal vein]. AB - OBJECTIVE: To evaluate three dimensional dynamic contrast-enhanced magnetic resonance angiography (3D-DCE MRA) in diagnosis of cavernous transformation of portal vein (CTPV). METHODS: Twenty-four patients with CTPV underwent 3D-DCE MRA examinations and the reconstructed images were retrospectively analyzed. A series of clinical, laboratory and imaging studies were performed on all these cases. Among all cases 14 underwent operations and 2 with hepatocellular carcinoma complicated portal thrombosis received transhepatic artery chemoembolization. RESULT: The CTPA was located in the main trunk in 10 cases, in both the main trunk and left/right branches in 8, and in left or right branches of the portal vein in 4. In the remaining 2 cases CTPA was located at the level of superior mesenteric vein. MRA revealed multiple circuitous collateral veins striding over obstruction to extend into the liver in 9 cases,and in 7 it simultaneously showed streaky or dot-like low signal intensities representing thrombi in the extensively dilated network of portal system. MRA did not clearly demonstrate the structure of the portal vein but only showed multiple sinuous network of venous collaterals strangling together in 6 cases. In 15 cases it also showed the route and distribution of multiple hepatofugal venous collaterals. CONCLUSION: 3D-DCE MRA can provide adequate information about the site and severity of CTPA. PMID- 18422285 TI - [Porous polyethylene channel implants in reconstruction of orbital blowout fractures]. AB - OBJECTIVE: To evaluate the efficacy of porous polyethylene channel implants in reconstruction of orbital blowout fractures. METHODS: Twenty-three patients with orbital blowout fractures were admitted in our eye center from June 2003 to January 2006. Porous polyethylene channel implants were used to repair the defects of orbital wall and to correct the enophthalmos. The visual acuity, the degree of enophthalmos, diplopia and position were recorded preoperatively and postoperatively. RESULT: No patients presented loss or apparent descent of visual acuity after surgery. Postoperative mean enophthalmos (1.7+/-0.5)mm was smaller significantly than that of preoperation [(3.9 +/-1.2)mm (P<0.05)]. Diplopia disappeared in 10 patients 1 month after surgery. Diplopia disappeared in 11 patients in primary position, but it remained in other positions. Diplopia remained in primary position in 2 patients. Ocular movement in 22 patients was normal postoperatively. While primary position upward occurred in 1 patient postoperatively and 2 weeks later it became normal. CONCLUSION: Porous polyethylene channel implants are effective in reconstruction of orbital blowout fractures. PMID- 18422286 TI - [Interaction of cancer-associated fibroblasts and tumor cells in carcinogenesis]. AB - OBJECTIVE: Tumor microenvironment is composed of mesenchymal cells and extracellular matrix components, which plays an important role in the growth, invasion and metastasis of tumor cells. Characteristic changes of stroma are usually accompanied with the malignant transformation of epithelial cells even ahead. The key component in the process-activated fibroblasts, also known as myofibroblasts, or cancer-associated fibroblasts (CAF), is the most abundant cells in tumor stroma. They promote the malignant transformation of epithelial cells through cell-cell communication via various soluble factors. This article reviews cancer-associated fibroblasts and their role in tumor development. PMID- 18422287 TI - [Percutaneous transluminal angioplasty for a patient with subclavian steal syndrome]. PMID- 18422288 TI - Asymmetric transfer hydrogenation of beta,gamma-alkynyl alpha-imino esters by a Bronsted acid. AB - Asymmetric synthesis of trans-alkenyl alpha-amino esters was realized by chiral phosphoric acid catalyzed transfer hydrogenation of beta,gamma-alkynyl alpha imino esters. Utilizing Hantzsch esters as the hydrogen donor, both the alkyne and imine moieties of beta,gamma-alkynyl alpha-imino esters were reduced to afford trans-alkenyl alpha-amino esters with up to 96% ee. PMID- 18422289 TI - Swallowtail bacteriochlorins. Lipophilic absorbers for the near-infrared. AB - Bacteriochlorins absorb strongly in the near-infrared spectral region and hence are ideally suited for diverse photomedical applications, yet naturally occurring bacteriochlorins have limited stability and synthetic malleability. A de novo route has been exploited to prepare synthetic bacteriochlorins that bear a geminal dimethyl group in each pyrroline ring for stability and a symmetrically branched 1,5-dimethoxypentyl group attached to each pyrrole ring for solubility in lipophilic media. PMID- 18422290 TI - Challenges in distinguishing superexchange and hopping mechanisms of intramolecular charge transfer through fluorene oligomers. AB - The temperature dependence of intramolecular charge separation in a series of donor-bridge-acceptor molecules having phenothiazine (PTZ) donors, 2,7 oligofluorene FL(n) (n = 1-4) bridges, and perylene-3,4:9,10-bis(dicarboximide) (PDI) acceptors was studied. Photoexcitation of PDI to its lowest excited singlet state results in oxidation of PTZ via the FL(n) bridge. In toluene, the temperature dependence of the charge separation rate constants for PTZ-FL(n)-PDI, (n = 1-4) is relatively weak and is successfully described by the semiclassical Marcus equation. The activation energies for charge separation suggest that bridge charge carrier injection is not the rate limiting step. The difficulty of using temperature and length dependence to differentiate hopping and superexchange is discussed, with difficulties in the latter topic explored via an extension of a kinetic model proposed by Bixon and Jortner. PMID- 18422291 TI - Hydration of OCS with one to four water molecules in atmospheric and laboratory conditions. AB - Carbonyl sulfide is the most abundant sulfur gas in the atmosphere. We have used MP2 and CCSD(T) theory to study the structures and thermochemistries of carbonyl sulfide interacting with one to four water molecules. We have completed an extensive search for clusters of OCS(H2O)n , where n = 1-4. We located three dimers, two trimers, five tetramers, and four pentamers with the MP2/aug-cc-pVDZ method. In each of the complexes with two or more waters, OCS preferentially interacts with low-energy water clusters. Our results match current theoretical and experimental literature, showing correlation with available geometries and frequencies for the OCS(H2O) species. The CCSD(T)/aug-cc-pVTZ thermochemical values combined with the average amount of OCS and the saturated concentration of H2O in the troposphere, lead to the prediction of 10(6) OCS(H2O) clusters x cm( 3) and 10(2) OCS(H2O)2 clusters x cm(-3) at 298 K. We predict the structures of OCS(H2O)n , n = 1-4 that should predominate in a low-temperature molecular beam and identify specific infrared vibrations that can be used to identify these different clusters. PMID- 18422292 TI - An ab initio study of the low-lying electronic states of YO2 and Franck-Condon simulation of the first photodetachment band of YO2(-). AB - A variety of density functional theory and ab initio methods, including B3LYP, B98, BP86, CASSCF, CASSCF/RS2, CASSCF/MRCI, BD, BD(T), and CCSD(T), with ECP basis sets of up to the quintuple-zeta quality for Y, have been employed to study the X(2)B2 state of YO2 and the X(1)A1 state of YO2(-). Providing that the Y 4s(2)4p(6) outer-core electrons are included in the correlation treatment, the RCCSD(T) method gives the most consistent results and is concluded to be the most reliable and practical computational method for YO2 and YO2(-). In addition, RCCSD(T) potential energy functions (PEFs) of the X(2)B2 state of YO2 and the X(1)A1 state of YO2(-) were computed, employing the ECP28MDF_aug-cc-pwCVTZ and aug-cc-pVTZ basis sets for Y and O, respectively. Franck-Condon factors, which include allowance for Duschinsky rotation and anharmonicity, were calculated using the computed RCCSD(T) PEFs and were used to simulate the first photodetachment band of YO2(-). The simulated spectrum matches very well with the corresponding experimental 355 nm photodetachment spectrum of Wu, H.; Wang, L.-S. J. Phys. Chem. A 1998, 102, 9129, confirming the reliability of the RCCSD(T) PEFs used. Further calculations on low-lying electronic states of YO2 gave T(e)'s and T(vert)'s of the A(2)A1, B(2)B1, and C(2)A2 states of YO2, as well as EAs and VDEs to these states from the X(1)A1 state of YO2(-). On the basis of the ab initio VDEs obtained in the present study, previous assignments of the second and third photodetachment bands of YO2(-) have been revised. PMID- 18422293 TI - Theoretical study of the nitroalkane thermolysis. 1. Computation of the formation enthalpy of the nitroalkanes, their isomers and radical products. AB - The gas phase enthalpies of formation of mono-, di-, tri-, tetranitromethane and nitroethane, as well as of their nitrite and aci-form isomers were calculated using different multilevel (G2, G3, G2M(CC5)) and density functional theory (DFT) based (B3LYP, MPW1B95 and MPWB1K) techniques. The enthalpies of the C-N bond dissociation and isomerization of these nitroalkanes were also calculated. The calculated values of the formation and reaction enthalpies were compared with the experimental data when these data were available. It was found that only the G3 procedure gave accurate (within 1 kcal/mol) results for the formation enthalpy of nitroalkanes, their isomers, and radical products. The G3 procedure and two new hybrid meta DFT methods proposed by Truhlar's group (Zhao, Y.; Truhlar, D. J. Phys. Chem. A 2004, 108, 6908) showed good results for the reaction enthalpies of the nitromethane isomerization and the C-N bond dissociation. Our calculation results were used to analyze thermodynamics of the dissociation and isomerization reactions of the poly nitro-substituted methanes. PMID- 18422294 TI - Electronic and vibrational spectroscopic investigation of phenylacetylene-amine complexes. Evidence for the diversity in the intermolecular structures. AB - Shifts in the electronic transitions for the complexes of phenylacetylene with ammonia, methylamine, and triethylamine clearly indicate the variation in the intermolecular structures of the three complexes. The infrared spectrum of phenylacetylene in the acetylenic C-H stretching region shows Fermi resonance bands, which act as a sensitive tool to probe the intermolecular structures. The IR-UV double resonance spectra of the three complexes are disparate and signify the formation of distinct structures. The formation of C-H...N hydrogen-bonded complex with ammonia and two distinct types of pi complexes with methylamine and triethylamine can be inferred from the analysis of electronic and vibrational spectra in combination with ab initio calculations. These complexes clearly point out the fact that marginal changes in the interacting partner can significantly alter the intermolecular structure. PMID- 18422295 TI - Equilibrium structure and torsional barrier of BH3NH3. AB - Born-Oppenheimer equilibrium structures, r(e)(BO), of the electronic ground state of the borazane (BH3NH3) molecule of C3v point-group symmetry are computed ab initio using the CCSD(T) method with basis sets up to quintuple-zeta quality. Inclusion of the counterpoise correction and extrapolation of the structural parameters to the complete basis set limit yield a best estimate of r(e)(BO) of BH3NH3. The anharmonic force field of BH3NH3, computed at the CCSD(T) level of theory with a basis set of triple-zeta quality, allows the determination of semi experimental equilibrium rotational constants, which in turn result in a semi experimental equilibrium structure, r(e)(SE). The r(e)(BO) and r(e)(SE) structures are in excellent agreement, indicating the validity of the methods used for their determination. The empirical mass-dependent structure, r(m)(1), of BH3NH3 is also determined. Although it is inferior in quality to the previous two structures, it is much more accurate than the standard empirical r0 and r(s) structures reported earlier for BH3NH3. The semi-experimental r(e)(SE) as well as the empirical r(m)(1) structures determined are based on experimental ground state rotational constants available from the literature for nine isotopologues of borazane. The effective barrier to the internal rotation of BH3NH3, a molecule isoelectronic with CH3CH3, has been computed ab initio, employing the focal-point analysis (FPA) approach, to be 699 +/- 11 cm(-1). This compares favorably with an empirical redetermination of the effective barrier based on the above r(e)(SE) structure, V3 = 718(17) cm(-1). PMID- 18422296 TI - Instituting greener chemistry. PMID- 18422300 TI - Joshua Coon: retooling chemical biology. PMID- 18422301 TI - Ester bonds in prodrugs. AB - A recent study challenges the oft-held notion that ester bonds in prodrug molecules are cleaved rapidly and completely inside cells by endogenous, nonspecific esterases. Structure-activity relationship studies on acylated sugars reveal that regioisomeric compounds display disparate biological activity, suggesting that ester bonds can persist in a cellular context. PMID- 18422302 TI - Allostery: DNA does it, too. AB - Allostery is a central concept for understanding protein function and regulation. It is less well appreciated that DNA is allosteric, too, and that DNA conformational changes can by coupled to protein binding interactions on the DNA lattice. Allosteric DNA interactions are emerging as important features in the assembly of the molecular machines that regulate transcription. PMID- 18422303 TI - Glucose: a novel regulator of notch signaling. AB - Notch signaling regulates cell fate during the development of many tissues. A new Drosophila mutant, rumi, is defective in Notch signaling because it cannot add glucose to serine in epidermal growth factor repeats of Notch extracellular domain. This is the first biological role for glucose covalently attached to a cell surface signaling receptor. PMID- 18422305 TI - Proteomics based on peptide fractionation by SDS-free PAGE. AB - Here we demonstrate the usefulness of peptide fractionation by SDS-free polyacrylamide gel electrophoresis and its applicability to proteomics studies. In the absence of SDS, the driving force for the electrophoretic migration toward the anode is supplied by negatively charged acidic amino acid residues and other residues as phosphate, sulfate and sialic acid, while the resulting mobility depends on both the charge and the molecular mass of the peptides. A straightforward method was achieved for SDS-PAGE of proteins, enzyme digestion, peptide transfer and fractionation by SDS-free PAGE, which was named dual fractionation polyacrylamide gel electrophoresis (DF-PAGE). This method increases the number of identified proteins 2.5-fold with respect to the proteins identified after direct analysis, and more than 80% of assigned peptides were found in unique SDS-free gel slices. A vast majority of identified peptides (93%) have p I values below 7.0, and 7% have p I values between 7.0 and 7.35. Peptide digests that were derived from complex protein mixtures were in consequence simplified as peptides that are positively charged are not recovered in the present conditions. The analysis of a membrane protein extract from Neisseria meningitidis by this approach allowed the identification of 97 proteins, including low-abundance components. PMID- 18422304 TI - Targeted chemical wedges reveal the role of allosteric DNA modulation in protein DNA assembly. AB - The cooperative assembly of multiprotein complexes results from allosteric modulations of DNA structure as well as direct intermolecular contacts between proteins. Such cooperative binding plays a critical role in imparting exquisite sequence specificity on the homeobox transcription factor (Hox) family of developmental transcription factors. A well-characterized example includes the interaction of Hox proteins with extradenticle (Exd), a highly conserved DNA binding transcription factor. Although direct interactions are important, the contribution of indirect interactions toward cooperative assembly of Hox and Exd remains unresolved. Here we use minor groove binding polyamides as structural wedges to induce perturbations at specific base steps within the Exd binding site. We find that allosteric modulation of DNA structure contributes nearly 1.5 kcal/mol to the binding of Exd to DNA, even in the absence of direct Hox contacts. In contrast to previous studies, the sequence-targeted chemical wedges reveal the role of DNA geometry in cooperative assembly of Hox-Exd complexes. Programmable polyamides may well serve as general probes to investigate the role of DNA modulation in the cooperative and highly specific assembly of other protein-DNA complexes. PMID- 18422306 TI - Synthesis and biological evaluation of pyridazinone analogues as potential cardiac positron emission tomography tracers. AB - A series of fluorinated pyridazinone derivatives with IC50 values ranging from 8 to 4000 nM for the mitochondrial complex 1 (MC1) have been prepared. Structure activity relationship (SAR) assessment indicated preference of the fluorine label to be incorporated on an alkyl side chain rather than directly on the pyridazinone moiety. Tissue distribution studies of a series of analogues ([18F] 22-28) in Sprague-Dawley (SD) rats identified [18F]27 as the most promising radiotracer with high uptake in cardiac tissue (3.41%ID/g; 30 min post injection) in addition to favorable heart to nontarget organ distribution ratios. MicroPET images of SD rats and nonhuman primates after [18F]27 administration allowed easy assessment of the myocardium through 60 min with minimal lung or liver interference. PMID- 18422307 TI - MM-GB/SA rescoring of docking poses in structure-based lead optimization. AB - The critical issues in docking include the prediction of the correct binding pose and the accurate estimation of the corresponding binding affinity. Different docking methodologies have all been successful in reproducing the crystallographic binding modes but struggle when predicting the corresponding binding affinities. The aim of this work is to evaluate the performance of the MM GB/SA rescoring of docking poses in structure-based lead optimization. To accomplish that, a diverse set of pharmaceutically relevant targets, including CDK2, FactorXa, Thrombin, and HIV-RT were selected. The correlation between the MM-GB/SA results and experimental data in all cases is remarkable. It even qualifies this approach as a more attractive alternative for rank-ordering than the Free Energy Perturbation and Thermodynamic Integration methodologies because, while as accurate, it can handle more structurally dissimilar ligands and provides results at a fraction of the computational cost. On the technical side, the benefit of performing a conformational analysis and having an ensemble of conformers to represent each ligand in the unbound state during the MM-GB/SA rescoring procedure was investigated. In addition, the estimation of conformational entropy penalties for the ligands upon binding, computed from the Boltzmann distribution in water, was evaluated and compared to a commonly used approach employed by many docking scoring functions. PMID- 18422308 TI - Cyclic (amino)[bis(ylide)]carbene as an anionic bidentate ligand for transition metal complexes. AB - The 3-(triphenylphosphonio)-N-(2,6-diisopropylphenyl)pyrrole reacts with 2 equiv of methyllithium to afford a lithium adduct in which a cyclic (amino)[bis(ylide)]carbene, a novel type of NHC, acts as a 1,4-bidentate ligand via the carbene center and the exocyclic ylidic carbon. This species readily undergoes transmetalation reactions, which allows for the synthesis of a variety of transition-metal complexes. PMID- 18422309 TI - A new class of luminescent tricarbonyl rhenium(I) complexes containing bridging 1,2-diazine ligands: electrochemical, photophysical, and computational characterization. AB - A novel class of luminescent tricarbonyl rhenium(I) complexes of general formula [Re2(mu-X)2(CO)6(mu-diaz)] (X=halogen and diaz=1,2-diazine) was prepared by reacting [ReX(CO)5] with 0.5 equiv of diazine (seven different ligands were used). The bridging coordination of the diazine in these dinuclear complexes was confirmed by single-crystal X-ray analysis. Cyclic voltammetry in acetonitrile showed for all the complexes (but the phthalazine derivative) a chemically and electrochemically reversible ligand-centered reduction, as well as a reversible metal-centered bielectronic oxidation. With respect to the prototypical luminescent [ReCl(CO)3(bpy)] complex, the oxidation is more difficult and the reduction easier (about +0.3 V), so that a similar highest occupied molecular orbital-lowest unoccupied molecular orbital gap is observed. All of the complexes exhibit photoluminescence at room temperature in solution, with broad unstructured emission from metal-to-ligand charge-transfer states, at lambda in the range 579-620 nm. Lifetimes (tau=20-2200 ns) and quantum yields (Phi up to 0.12) dramatically change upon varying the bridging ligand X and the diazine substituents: in particular, quantum yields decrease in the series Cl, Br, and I and in the presence of substituents at the alpha positions of the pyridazine ring. A combined density functional and time-dependent density functional study of the geometry, relative stability, electronic structure, and photophysical properties of all the pyridazine derivatives was performed. The nature of the excited states involved in the electronic absorption spectra was ascertained, and trends in the energy of the highest occupied and lowest unoccupied molecular orbitals upon changing the pyridazine substituents and the bridging halogen ligands were discussed. The observed emission properties of these complexes were shown to be related to a combination of steric and electronic factors affecting their ground-state geometry and their stability. PMID- 18422310 TI - A two-dimensional iron(II) carboxylate linear chain polymer that exhibits a metamagnetic spin-canted antiferromagnetic to single-chain magnetic transition. AB - A two-dimensional iron(II) carboxylate coordination polymer, [Fe(pyoa)2]infinity, where pyoa is 2-(pyridin-3-yloxy)acetate, has been prepared by hydrothermal synthesis. Its crystal structure reveals a single iron(II) site with an elongated octahedral coordination environment containing four equatorial carboxylate oxygens and two axial pyridyl nitrogens; the iron(II) sites are linked by syn anti micro-carboxylates to form chains along the b axis that have an Fe...Fe separation of 4.910 A. The shortest interchain and interlayer Fe...Fe distances are 6.453 and 11.125 A, respectively. The 4.2-295 K Mossbauer spectra of [Fe(pyoa) 2] infinity consist of a single paramagnetic high-spin iron(II) quadrupole doublet. The axial Fe-N bond direction defines the Jahn-Teller axis at an iron(II) site and, consequently, the orientation of the single-ion magnetic anisotropy. Thus, along the b axis in a given chain, the spins are collinear and parallel to the Jahn-Teller axis. The Jahn-Teller axes of adjacent intralayer chains have different orientations with an angle of 79.2 degrees between the axes in adjacent chains in a bc layer. [Fe(pyoa)2]infinity exhibits field-induced metamagnetic behavior such that, in an applied field smaller than the critical field, the iron(II) spin-canted moments experience intrachain ferromagnetic interactions and weak interchain antiferromagnetic interactions; the spin canting yields weak ferromagnetism. In an applied field larger than the critical field, the weak antiferromagnetic interchain interactions are overwhelmed to yield superparamagnetic-like slow-magnetic relaxation with an energy barrier of 23(3) K. Single-crystal magnetic studies reveal a quasi-uniaxial magnetic anisotropy with the a axis as the easy-magnetic axis and the b axis as the hard-magnetic axis; the susceptibility measured along the easy a axis may be fit with the Glauber model to yield an effective intrachain exchange coupling constant of 2.06(8) K. A dynamic analysis of the susceptibility yields a 6.3(1) K energy barrier for intrachain domain wall creation. The observed field-assisted superparamagnet-like behavior is consistent with the dynamics of a single-chain magnet. Thus, [Fe(pyoa)2]infinity is best considered as a "metamagnetic-like" single-chain magnet. PMID- 18422311 TI - Ligand localized triplet excited states in platinum(II) bipyridyl and terpyridyl peryleneacetylides. AB - An investigation of the photophysics of two complexes, [Pt((t)Bu3tpy)(C triple bond C-perylene)]BF4 (1) and Pt((t)Bu2bpy)(C triple bond C-perylene)2 (2), where (t)Bu3tpy is 4,4',4''-tri( tert-butyl)-2,2':6',2''-terpyridine, (t)Bu2bpy is 4,4' di( tert-butyl)-2,2'-bipyridine, and C triple bond C-perylene is 3 ethynylperylene, reveals that they both exhibit perylene-centered ligand localized excited triplet states ((3)IL) upon excitation with visible light. These complexes do not display any significant photoluminescence at room temperature but readily sensitize (1)O2 in aerated CH2Cl2 solutions, as evidenced by its characteristic emission near 1270 nm. The transient absorption difference spectra were compared to bi- and tridentate phosphine peryleneacetylides intended to model the (3)IL peryleneacetylide excited states in addition to the related phenylacetylide-bearing polyimine analogues, with the latter model being the respective triplet charge-transfer ((3)CT) excited states. The transient difference spectra of the two title compounds display excited-state absorptions largely attributable to perylene localized (3)IL states yet exhibit somewhat attenuated excited-state lifetimes relative to those of the phosphine model chromophores. The abbreviated lifetimes in 1 and 2 may suggest the involvement of the energetically proximate (3)CT triplet state exerting an influence on excited state decay, and the effect appears to be stronger in 1 relative to 2, consistent with the energies of their respective (3)CT states. PMID- 18422312 TI - Interaction of molecular hydrogen with open transition metal centers for enhanced binding in metal-organic frameworks: a computational study. AB - Molecular hydrogen is known to form stable, "nonclassical" sigma complexes with transition metal centers that are stabilized by donor-acceptor interactions and electrostatics. In this computational study, we establish that strong H2 sorption sites can be obtained in metal-organic frameworks by incorporating open transition metal sites on the organic linkers. Using density functional theory and energy decomposition analysis, we investigate the nature and characteristics of the H2 interaction with models of exposed open metal binding sites {half sandwich piano-stool shaped complexes of the form (Arene)ML(3- n)(H2)n [M=Cr, Mo, V(-), Mn(+); Arene = C6H5X (X=H, F, Cl, OCH3, NH2, CH3, CF3) or C6H3Y2X (Y=COOH, X=CF3, Cl; L=CO; n=1-3]}. The metal-H2 bond dissociation energy of the studied complexes is calculated to be between 48 and 84 kJ/mol, based on the introduction of arene substituents, changes to the metal core, and of charge-balancing ligands. Thus, design of the binding site controls the H2 binding affinity and could be potentially used to control the magnitude of the H2 interaction energy to achieve reversible sorption characteristics at ambient conditions. Energy decomposition analysis illuminates both the possibilities and present challenges associated with rational materials design. PMID- 18422314 TI - Catalytic asymmetric epoxidation of cyclic enones. AB - A highly enantioselective epoxidation of cyclic enones with hydrogen peroxide has been developed that is catalyzed by chiral primary amine salts. PMID- 18422315 TI - The isolable cation radical of disilene: synthesis, characterization, and a reversible one-electron redox system. AB - The highly twisted tetrakis(di-tert-butylmethylsilyl)disilene 1 was treated with Ph3C+.BAr4- (BAr4-: TPFPB = tetrakis(pentafluorophenyl)borate) in toluene, producing disilene cation radical 3 upon one-electron oxidation. Cation radical 3 was isolated in the form of its borate salt as extremely air- and moisture sensitive red-brown crystals. The molecular structure of 3 was established by X ray crystallography, which showed a highly twisted structure (twisting angle of 64.9 degrees) along the central Si-Si bond with a bond length of 2.307(2) A, which is 2.1% elongated relative to that of 1. The cation radical is stabilized by sigma-pi hyperconjugation by the four tBu2MeSi groups attached to the two central sp2-Si atoms. An electron paramagnetic resonance (EPR) study of the hyperfine coupling constants (hfcc) of the 29Si nuclei indicates delocalization of the spin over the central two Si atoms. A reversible one-electron redox system between disilene, cation radical, and anion radical is also reported. PMID- 18422313 TI - Metal-mediated self-assembly of protein superstructures: influence of secondary interactions on protein oligomerization and aggregation. AB - We have previously demonstrated that non-self-associating protein building blocks can oligomerize to form discrete supramolecular assemblies under the control of metal coordination. We show here that secondary interactions (salt bridges and hydrogen bonds) can be critical in guiding the metal-induced self-assembly of proteins. Crystallographic and hydrodynamic measurements on appropriately engineered cytochrome cb562 variants pinpoint the importance of a single salt bridging arginine side chain in determining whether the protein monomers form a discrete Zn-induced tetrameric complex or heterogeneous aggregates. The combined ability to direct PPIs through metal coordination and secondary interactions should provide the specificity required for the construction of complex protein superstructures and the selective control of cellular processes that involve protein-protein association reactions. PMID- 18422316 TI - Identifying the minimal enzymes required for anhydrotetracycline biosynthesis. AB - The cyclohexenone ring A of tetracyclines exhibits unique structural features not observed among other aromatic polyketides. These substitutions include the C2 primary amide, C4 dimethylamine, and the C12a tertiary alcohol. Here we report the identification and reconstitution of the minimum set of enzymes required for the biosynthesis of anhydrotetracycline (ATC, 5), the first intermediate in the tetracycline biosynthetic pathway that contains the fully functionalized ring A. Using a combination of in vivo and in vitro approaches, we confirmed OxyL, OxyQ, and OxyT to be the only enzymes required to convert 6-methylpretetramid 1 into 5. OxyL is a NADPH-dependent dioxygenase that introduces two oxygen atoms into 1 to yield the unstable intermediate 4-keto-ATC 2. The aminotransferase OxyQ catalyzes the reductive amination of C4-keto of 2, yielding 4-amino-ATC 3. Furthermore, the N, N-dimethyltransferase OxyT catalyzes the formation of 5 from 3 in a (S) adenosylmethionine (SAM)-dependent manner. Finally, a "non-natural" anhydrotetracycline derivative was generated, demonstrating that our heterologous host/vector pair can be a useful platform toward the engineered biosynthesis of tetracycline analogues. PMID- 18422317 TI - Thiolate bridging and metal exchange in adducts of a zinc finger model and Pt(II) complexes: biomimetic studies of protein/Pt/DNA interactions. AB - To provide precedents for the possible interactions of platinum DNA adducts with zinc finger proteins, the complexes [Pt(dien)Cl]Cl (dien = diethylenetriamine) and [Pt(terpy)Cl]Cl (terpy = 2,2':6',2''-terpyridine) were exposed to the N,N' bis(2-mercaptoethyl)-1,4-diazacycloheptanezinc(II) dimer, [Zn(bme-dach)]2, and the products defined by electrospray ionization mass spectrometry (ESI-MS), X-ray crystallography and (195)Pt NMR spectroscopy. The presence of a leaving chloride in both platinum(II) complexes facilitates electrophilic substitution involving sulfur-containing zinc finger synthetic models or, as in previous studies, zinc finger peptidic sequences. Monitored via ESI-MS, both reactants yielded evidence for Zn-(mu-SR)-Pt bridges followed by zinc ejection from the N2S2 coordination sphere and subsequent formation of a trimetallic Zn-(mu-SR)2-Pt-(mu-SR)2-Zn bridged species. The isolation of Zn-(mu-SR)-Pt-bridged species [(Zn(bme dach)Cl)(Pt(dien))]Cl is, to our knowledge, the first Zn-Pt bimetallic thiolate bridged model demonstrating the interaction between Zn-bound thiolates and Pt(2+). In the case of the [Pt(terpy)Cl]Cl reaction with the [Zn(bme-dach)]2, ESI MS analysis further suggests metal exchange by formation of [Zn(terpy)Cl](+), whereas the [Pt(dien)Cl]Cl reaction does not yield the corresponding [Zn(dien)Cl](+) ion. Direct synthesis of the Zn-Pt thiolate-bridged species and the Pt(N2S2) chelate, where Pt has displaced the Zn from the chelate core, permitted the isolation of X-ray-quality crystals to confirm the bridging and metal-exchanged structures. The ESI-MS, (195)Pt NMR spectroscopy, and molecular structures of the di- and trinuclear complexes will be discussed, as they provide insight into the metal-exchange mechanism. PMID- 18422319 TI - Concerning the synthesis of the tedanolide C(13)-C(23) fragment via anti-aldol reaction. AB - Synthesis of C(13)-C(23) aldehyde 4, an important intermediate in a planned total synthesis of tedanolide, is described. The stereoselectivity of the key anti aldol reaction of aldehyde 5 and ketone 6 (en route to 4) perfectly tracks the enantiomeric purity of 5. It is demonstrated that aldehyde 24, a precursor of 5, undergoes facile epimerization during a Swern oxidation and stabilized ylide olefination sequence. PMID- 18422318 TI - Role of hydrogen bonding and helix-lipid interactions in transmembrane helix association. AB - To explore the role of hydrogen bonding and helix-lipid interactions in transmembrane helix association, we have calculated the potential of mean force (PMF) as a function of helix-helix distance between two pVNVV peptides, a transmembrane model peptide based on the GCN4 leucine-zipper, in a dimyristoylphosphatidylcholine (DMPC) membrane. The peptide name pVNVV represents the interfacial residues in the heptad repeat of the dimer. The free energy decomposition reveals that the total PMF consists of two competing contributions from helix-helix and helix-lipid interactions. The direct, favorable helix-helix interactions arise from the specific contribution from the helix-facing residues and the generic contribution from the lipid-facing residues. The Asn residues in the middle of the helices show the most significant per-residue contribution to the PMF with various hydrogen bonding patterns as a function of helix-helix distance. Release of lipid molecules between the helices into bulk lipid upon helix association makes the helix-lipid interaction enthalpically unfavorable but entropically favorable. Interestingly, the resulting unfavorable helix-lipid contribution to the PMF correlates well with the cavity volume between the helices. The calculated PMF with an Asn-to-Val mutant (pVNVV --> pVVVV) shows a dramatic free energy change upon the mutation, such that the mutant appears not to form a stable dimer below a certain peptide concentration, which is in good agreement with available experimental data of a peptide with the same heptad repeat. A transmembrane helix association mechanism and its implications in membrane protein folding are also discussed. PMID- 18422320 TI - Gassman's intramolecular [2 + 2] cationic cycloaddition. formal total syntheses of raikovenal and epi-raikovenal. AB - The first intramolecular version of Gassman's cationic [2 + 2] cycloaddition employing vinyl acetals tethered to an unactivated olefin and its application in the formal syntheses of raikovenal and epi-raikovenal are described. PMID- 18422321 TI - Tandem regio- and stereoselective organocuprate-mediated bis-allylic substitutions. AB - anti-5-Acetoxy-4-halo-alpha,beta-enoates undergo sequential or tandem reactions with two different magnesium cuprate reagents to afford anti-2,3-dialkyl-4,5 enoates in high chemical yield and with excellent diastereoselectivity. The one pot tandem procedure can be achieved with 30 mol % of CuCN and affords a rapid stereoselective combinatorial approach to vicinal disubstituted gamma,delta enoates containing functionality at either end of the carbon chain for subsequent functional group elaboration. The methodology should provide a powerful practical strategy for acyclic stereoselection. PMID- 18422322 TI - Biphenylene-substituted ruthenocenylphosphine for Suzuki-Miyaura coupling of aryl chlorides. AB - High activity in the palladium-catalyzed Suzuki-Miyaura reactions of aryl chlorides with arylboronic acids was furnished using biphenylene-substituted di- tert-butylruthenocenylphosphine (R-Phos) as a supporting ligand. Substrate combinations even for the construction of highly hindered tetra- ortho substituted biaryls can be achieved in good to excellent yields with low catalyst loadings in short reaction times. PMID- 18422323 TI - Michael addition of ortho-lithiated aryloxiranes to alpha,beta-unsaturated malonates: synthesis of tetrahydroindenofuranones. AB - A short and efficient synthesis of tetrahydroindenofuranones based on the Michael addition of ortho-lithiated aryloxiranes to alkylidene malonates followed by the nucleophilic oxirane ring-opening and subsequent lactonization is described. The methodology has been applied to the synthesis of a structural analogue of epipodophyllotoxins. PMID- 18422324 TI - Copper-catalyzed synthesis of esters from ketones. Alkyl group as a leaving group. AB - The conversion of ketones to esters has been achieved through the use of Cu catalyst and tetrabutylammonium nitrite. This reaction involves the activation of the less activated C-C bond, and the alkyl group is removed as a leaving group. Various isopropyl ketones are found to be good substrates for this reaction. PMID- 18422325 TI - A highly effective cobalt catalyst for olefin aziridination with azides: hydrogen bonding guided catalyst design. AB - [Co(P1)], which was designed on the basis of potential hydrogen-bonding interactions in the metal-nitrene intermediate, is a highly active aziridination catalyst with azides. [Co(P1)] can effectively aziridinate various aromatic olefins with arylsulfonyl azides under mild conditions, forming sulfonylated aziridines in excellent yields. The Co-based system enjoys several attributes associated with the relatively low cost of cobalt and the wide accessibility of arylsulfonyl azides. Furthermore, it generates stable dinitrogen as the only byproduct. PMID- 18422326 TI - Formation of protein-oligosaccharide conjugates by laccase and tyrosinase. AB - Proteins and certain carbohydrates contain phenolic moieties, which are potential sites for modification of the function of the biopolymers. In this study, the capability of two different fungal oxidative enzymes, laccase from Trametes hirsuta (ThL) and tyrosinase from Trichoderma reesei (TrT), to catalyze formation of hetero-cross-linking between tyrosine side chains of alpha-casein and phenolic acids of hydrolyzed oat spelt xylan (hOSX) was studied. Formation of reaction products was followed by size exclusion chromatography (SEC), fluorescence spectroscopy, and SDS-PAGE, using specific staining methods for proteins and protein-carbohydrate conjugates. ThL and TrT were observed to differ significantly in their ability to catalyze the formation of protein-carbohydrate conjugates or the linking of the small molecular weight phenolic compounds to alpha-casein. The efficiency of these enzymes to directly cross-link protein also differed notably. TrT was able to cross-link alpha-casein more efficiently than ThL. ThL-catalyzed casein cross-linking was significantly enhanced by ferulic acid, p-coumaric acid, and also hOSX. The main reaction products by ThL appeared to be phenolic acid-bridged alpha-caseins. Indications of hetero-cross-link formation between alpha-casein and hOSX by both oxidative enzymes could be visualized by glycoprotein-specific staining in the SDS-PAGE analysis, although ThL was observed to be more effective in the heteroconjugate formation than TrT. PMID- 18422327 TI - Changes in volatile compounds and overall aroma profile during storage of coffee brews at 4 and 25 degrees C. AB - In this work, the chemical changes occurring in the volatile fraction of Arabica coffee brews during storage at 4 and 25 degrees C for 30 days have been characterized for the first time by means of HS-GC-MS. A total of 47 compounds were identified and quantified: 2 sulfur compounds, 7 aldehydes, 3 esters, 15 furans, 5 ketones, 1 alcohol, 2 thiophenes, 4 pyrroles, 1 pyridine, 5 pyrazines, 1 alkene, and 1 acid. No new volatile compounds were detected at the end of the storage time. The changes observed are, in general, slower and less pronounced at refrigeration temperature. Storage also affects the sensory characteristics of the stored coffee brews, which lose part of their aroma intensity and freshness, acquiring some nondesirable notes such as rancid aroma, mainly during storage at 25 degrees C. Furthermore, seven aroma indices have been proposed as indicators of coffee brew staling, which show a good correlation with some sensory attributes, not only for aroma but also overall sensory quality. Consequently, they could be considered useful to monitor both the "age" and the sensory quality of stored coffee brews. PMID- 18422328 TI - Cytotoxic phenylpropanoids from carrot. AB - Carrot is widely used as a foodstuff. The active components such as beta-carotene and panaxynol have been studied by many researchers. In this investigation of nonpolar active components from carrot, a new phenylpropanoid, epilaserine oxide ( 3), was isolated along with six known compounds, laserine ( 1), 2-epilaserine ( 2), panaxynol ( 4), ginsenoyne K ( 5), (8 E)-1,8-heptadecadiene-4,6-diyne-3,10 diol ( 6), and vaginatin ( 7). Their structures were deduced on the basis of spectroscopic methods. Significant cytotoxicity of 2-epilaserine against HL-60 cells was observed, which implied that phenylpropanoids were cytotoxic compounds in carrot. Laserine and 2-epilaserine in carrots from diverse locations in China were quantified by HPLC. PMID- 18422329 TI - Extremely low temperature fermentations of grape must by potato-supported yeast, strain AXAZ-1. A contribution is performed for catalysis of alcoholic fermentation. AB - This investigation announces the use of potato pieces as a suitable support for cell immobilization resulting in extremely low temperature wine making. The results showed an increase of the total esters by immobilized cells and reduction of higher alcohols. Likewise, percentages of total esters on total volatiles were increased by the drop in temperature, while percentages of higher alcohols were reduced in wines. Kinetics experiments at different temperatures allowed the calculation of activation energy (Ea) and showed reduction in the case of immobilized cells as compared with free cells. These results may lead to the conclusion that the increased productivities that are obtained by immobilized cells, can be attributed to the catalytic activity by the support to enzymes, which are involved in the process. Biocatalysts were prepared by immobilization of Saccharomyces cerevisiae, strain AXAZ-1, on whole potatoes and potato pieces, and their efficiency for alcoholic repeated batch fermentations of glucose and grape must in the range 2-30 degrees C was examined. To study the operational stability of biocatalyst, 35 repeated batch fermentations of grape must were performed without any significant reduction of the fermentation activity. Wines were analyzed for volatile byproducts determination by GC and GC-MS. PMID- 18422330 TI - Biofuels: thinking clearly about the issues. AB - Dr. Bruce Dale is the recipient of the 2007 Sterling B. Hendricks Memorial Lectureship Award. This perspective is based on a lecture given by Dr. Dale at the Life Sciences and Society Symposium at the University of Missouri on March 14, 2007, and the Award Address he presented for the Sterling B. Hendricks Memorial Lectureship Award at the 234th National Meeting of the American Chemical Society, in Boston, MA, on August 20, 2007. PMID- 18422331 TI - Antioxidant and iron-binding properties of curcumin, capsaicin, and S allylcysteine reduce oxidative stress in rat brain homogenate. AB - Research demonstrates that antioxidants and metal chelators may be of beneficial use in the treatment of neurodegenerative diseases, such as Alzheimer's disease (AD). This study investigated the antioxidant and metal-binding properties of curcumin, capsaicin, and S-allylcysteine, which are major components found in commonly used dietary spice ingredients turmeric, chilli, and garlic, respectively. The DPPH assay demonstrates that these compounds readily scavenge free radicals. These compounds significantly curtail iron- (Fe2+) and quinolinic acid (QA)-induced lipid peroxidation and potently scavenge the superoxide anion generated by 1 mM cyanide in rat brain homogenate. The ferrozine assay was used to measure the extent of Fe2+ chelation, and electrochemistry was employed to measure the Fe3+ binding activity of curcumin, capsaicin, and S-allylcysteine. Both assays demonstrate that these compounds bind Fe2+ and Fe3+ and prevent the redox cycling of iron, suggesting that this may be an additional method through which these agents reduce Fe2+-induced lipid peroxidation. This study demonstrates the antioxidant and metal-binding properties of these spice ingredients, and it is hereby postulate that these compounds have important implications in the prevention or treatment of neurodegenerative diseases such as AD. PMID- 18422332 TI - Influence of time, surface-to-volume ratio, and heating process (continuous or intermittent) on the emission rates of selected carbonyl compounds during thermal oxidation of palm and soybean oils. AB - The aim of this work was to compare the emission rates of selected carbonyl compounds (CC) produced by palm and soybean oils when heated at 180 degrees C in the presence of air, through different time intervals and at different surface-to volume ratios ( S/ V), in continuous and intermittent processes. The CC were collected and derivatized onto silica C18 cartridges impregnated with an acid 2,4 dinitrophenylhidrazine solution, followed by extraction with acetonitrile and analysis by HPLC-UV and, in some cases, HPLC-MS with electrospray ionization. Among the CC quantified, namely, acetaldehyde, acrolein, propanal, butanal, hexanal, 2-heptenal, and 2-octenal, acrolein was the main emission in both oils and all S/ V ratios, followed by hexanal and 2-heptenal. The soybean oil has presented greater emission rates of acrolein than palm oil. When different S/ V ratios used during the heating process of the oil were compared, the emission rates, in general, were directly related to them, although saturated and nonsaturated CC have had different behaviors toward oxidation reactions. During intermittent heating, there was a trend of increasing emission rates of saturated aldehydes, whereas the opposite was observed with unsaturated aldehydes, probably due to the reactivity of the double bond present in these compounds. PMID- 18422333 TI - High processing tolerances of immunomodulatory proteins in Enoki and Reishi mushrooms. AB - This study investigated the processing tolerances of two mushroom proteins with immunomodulatory activities, including FVE from Enoki ( Flammulia velutipes ) and LZ8 from Reishi ( Ganoderma lucidum ) mushrooms, under food processing treatments such as heating, sterilization, frozen storage, extraction in acid/alkaline conditions, and dehydration. Results showed that the capability of these two proteins to induce IFN-gamma secretion by murine splenocytes remained after 100 degrees C heating for 30 min, 121 degrees C autoclaving for 15 min, and -80 degrees C freezing. The retained activities of both proteins on cell proliferation and IFN-gamma production did not decrease at 0.6 M hydrochloric acid (at pH 2) but strikingly dropped at 5 M sodium hydrate (at pH 13). After vacuum dehydration, FVE and LZ8 retained most of their activities on cell proliferation; nevertheless, the IFN-gamma secretion decreased to about half of the initial values. These findings suggest that these two mushroom proteins have a good thermal/freezing resistance, acid tolerance, and dehydration stability and are candidates for processing in food and nutraceutical utilization. PMID- 18422334 TI - Polyphenolic transmission to Segureno lamb meat from ewes' diet supplemented with the distillate from rosemary (Rosmarinus officinalis) leaves. AB - The aim of the present work is to study whether the introduction of rosemary plant byproduct, from plant steam distillation, in daily Segurena sheep feeding allows the transfer of active antioxidant components to lamb meat, without detriment to the animal productivity. For this, 36 Segurena ewes were assigned randomly to three homogeneous groups. One group was fed a basal diet as a control and the diet of the other two groups was modified by substituting 10 or 20% of the control diet (respectively) with distilled rosemary leaves. Chromatographic analysis allowed the identification of 11 polyphenolic components previously identified in the rosemary and basal diet pellets, respectively. Among them, rosmarinic acid, carnosol, and carnosic acid were the phenolic components that had a significantly increased presence ( P < 0.05) in the lamb meat from sheep mothers fed this aromatic herb, when compared to the control group. The incorporation of this byproduct into the animal diet favored the antioxidant capacity of these lamb meat samples. Fresh meat produced on rosemary had higher total ferric reducing antioxidant power (FRAP) ( P < 0.05), greater ability to reduce ABTS*+, and lower IC50 (DPPH*) ( P < 0.05) values when compared to the control group. Because no statistically significant differences were detected among the results obtained from the lamb meat belonging to the different ewe groups fed rosemary leaf extract (10 or 20%), it can be concluded that the incorporation of distilled rosemary leaves at a rate of 10% of the ewes' diet should be enough to improve the lamb meat antioxidant status. PMID- 18422335 TI - Geographical characterization of greek virgin olive oils (cv. Koroneiki) using 1H and 31P NMR fingerprinting with canonical discriminant analysis and classification binary trees. AB - This work deals with the prediction of the geographical origin of monovarietal virgin olive oil (cv. Koroneiki) samples from three regions of southern Greece, namely, Peloponnesus, Crete, and Zakynthos, and collected in five harvesting years (2001-2006). All samples were chemically analyzed by means of 1H and 31P NMR spectroscopy and characterized according to their content in fatty acids, phenolics, diacylglycerols, total free sterols, free acidity, and iodine number. Biostatistical analysis showed that the fruiting pattern of the olive tree complicates the geographical separation of oil samples and the selection of significant chemical compounds. In this way the inclusion of the harvesting year improved the classification of samples, but increased the dimensionality of the data. Discriminant analysis showed that the geographical prediction at the level of three regions is very high (87%) and becomes (74%) when we pass to the thinner level of six sites (Chania, Sitia, and Heraklion in Crete; Lakonia and Messinia in Peloponnesus; Zakynthos). The use of classification and binary trees made possible the construction of a geographical prediction algorithm for unknown samples in a self-improvement fashion, which can be readily extended to other varieties and areas. PMID- 18422336 TI - Optical anisotropy of supported lipid structures probed by waveguide spectroscopy and its application to study of supported lipid bilayer formation kinetics. AB - Supramolecular conformation and molecular orientation was monitored during supported lipid bilayer (SLB) formation using dual polarization interferometry (DPI). DPI was shown to enable real time sensitive determination of birefringence of the lipid bilayer together with thickness or refractive index (with the other a fixed value). This approach removes differences in mass loading due to anisotropy, so the mass becomes solely a function of the lipid d n/d c value. DPI measurements show highly reproducible qualitative and quantitative results for adsorption of liposomes of different lipid compositions and in buffers with or without CaCl 2. The packing of solvent-free self-assembled SLBs is shown to differ from other preparation methods. Birefringence analysis accompanied by mass and thickness measurements shows characteristic features of vesicle adsorption and SLB formation kinetics previously not demonstrated by evanescent optical techniques, including indications of percolation-type rupture of clusters of liposomes on the surface and correlated adsorption kinetics induced by liposome charge repulsion. Our study demonstrates that understanding of mechanistic details for an adsorption process for which conformational changes and ordering occur can be elucidated using DPI and greatly enhanced by modeling of optical birefringence. The data is in some respects more detailed than what can be obtained with conventional biosensing techniques like surface plasmon resonance and complementary to methods such as the quartz crystal microbalance. PMID- 18422337 TI - Subattomole sensitivity in biological accelerator mass spectrometry. AB - The Uppsala University 5 MV Pelletron tandem accelerator has been used to study (14)C-labeled biological samples utilizing accelerator mass spectrometry (AMS) technology. We have adapted a sample preparation method for small biological samples down to a few tens of micrograms of carbon, involving among others, miniaturizing of the graphitization reactor. Standard AMS requires about 1 mg of carbon with a limit of quantitation of about 10 amol. Results are presented for a range of small sample sizes with concentrations down to below 1 pM of a pharmaceutical substance in human blood. It is shown that (14)C-labeled molecular markers can be routinely measured from the femtomole range down to a few hundred zeptomole (10 (-21) mol), without the use of any additional separation methods. PMID- 18422338 TI - Web server based complex mixture analysis by NMR. AB - Comprehensive metabolite identification and quantification of complex biological mixtures are central aspects of metabolomics. NMR shows excellent promise for these tasks. An automated fingerprinting strategy is presented, termed COLMAR query, which screens NMR chemical shift lists or raw 1D NMR cross sections taken from covariance total correlation spectroscopy (TOCSY) spectra or other multidimensional NMR spectra against an NMR spectral database. Cross peaks are selected using local clustering to avoid ambiguities between chemical shifts and scalar J-coupling effects. With the use of three different algorithms, the corresponding chemical shift list is then screened against chemical shift lists extracted from 1D spectra of a NMR database. The resulting query scores produced by forward assignment, reverse assignment, and bipartite weighted-matching algorithms are combined into a consensus score, which provides a robust means for identifying the correct compound. The approach is demonstrated for a metabolite model mixture that is screened against the metabolomics BioMagResDatabank (BMRB). This NMR-based compound identification approach has been implemented in a public Web server that allows the efficient analysis of a wide range of metabolite mixtures. PMID- 18422339 TI - Green copper pigments biodegradation in cultural heritage: from malachite to moolooite, thermodynamic modeling, X-ray fluorescence, and Raman evidence. AB - Moolooite (copper oxalate), a very rare compound, was found as a degradation product from the decay of malachite in several specimens of Cultural Heritage studied. Computer simulations, based on heterogeneous chemical equilibria, support the transformation of malachite to moolooite through the intermediate copper basic sulfates or copper basic chlorides, depending on the presence of available free sulfate or chloride anions in the chemical system. Raman and X-ray fluorescence spectral evidence found during the analysis of the three case studies investigated supported the model predictions. According to the study, the presence of lichens and other microorganisms might be responsible for the decay phenomena. This work tries to highlight the importance of biological attack on specimens belonging to Cultural Heritage and to demonstrate the consequences of oxalic acid, excreted by some of these microorganisms, on the conservation and preservation of artwork. PMID- 18422340 TI - Poly(dimethylsiloxane) as passive sampler material for hydrophobic chemicals: effect of chemical properties and sampler characteristics on partitioning and equilibration times. AB - Information about sampling rates and equilibration times of passive samplers is essential in their calibration in field monitoring studies as well as sorption studies. The kinetics of a sampler depends on the distribution coefficient between the sampler material and aqueous phase and the exchange rates of chemicals between these phases. In this study, the elimination kinetics of four poly(dimethylsiloxane) (PDMS) passive samplers with different surface-volume ratios are compared. The samplers were loaded with polychlorinated biphenyls (PCBs) and polybrominated diphenyl ethers (PBDEs) that cover a broad range of hydrophobicities. The surface-volume ratios of the samplers could largely explain the observed kinetics. Furthermore, a simple diffusion-based model illustrates that the exchange of chemicals was limited by diffusion through the aqueous diffusion layer surrounding the sampler. On the basis of this simple diffusion model, equilibration times are predicted for organic chemicals that vary in hydrophobicity and samplers with different dimensions and polymeric phases. This information is of importance in the selection of a passive sampler for a specific purpose. PMID- 18422341 TI - High-throughput small angle X-ray scattering from proteins in solution using a microfluidic front-end. AB - This manuscript presents, for the first time, the method of automated structural analysis of biomolecules in solution on a microfluidic chip. A polymer-based micrototal analysis system for high-throughput Small-Angle X-ray Scattering (SAXS) data collection from biological macromolecules has been developed. The bioXTAS chip features an integrated X-ray transparent 200 nL sample chamber and diffusion-based mixing of protein and buffer solutions. Software for fully automated fluidic control, data acquisition, and data analysis has been developed. The proof-of concept is based on data using bovine serum albumin as the model system. It confirms the quality of SAXS data generated from small sample volumes and furthermore validates the on-chip mixing capabilities. SAXS data on the gradual unfolding of BSA induced by an anionic surfactant exemplifies how the bioXTAS chip can be used to follow and identify structural changes and proves the feasibility of high-throughput structural analysis in solution. In total, this shows that the bioXTAS chip has the potential for becoming a powerful tool for automated high-throughput structural analysis of macromolecular systems. PMID- 18422342 TI - Structure of the covalent adduct formed between Mycobacterium tuberculosis beta lactamase and clavulanate. AB - The intrinsic resistance of Mycobacterium tuberculosis to the beta-lactam class of antibiotics arises from a chromosomally encoded, extended spectrum, class A beta-lactamase, BlaC. Herein, we report the X-ray crystallographic structure of BlaC inhibited with clavulanate at a resolution of 1.7 A with an R-factor value of 0.180 and R-free value of 0.212 for the m/ z +154 clavulanate-derived fragment observed in the active site. Structural evidence reveals the presence of hydrogen bonds to the C1 carbonyl along with a coplanar arrangement of C1, C2, C3, and N4, which favors enolization to generate a trans-alpha,beta-eneamine, stabilizing the +154 adduct from hydrolysis. The irreversible inhibition of BlaC suggests that treatment of M. tuberculosis with a combination of a beta-lactam antibiotic and clavulanate may lead to rapid bactericidal activity. PMID- 18422344 TI - Chemical and steady-state kinetic analyses of a heterologously expressed heme dependent chlorite dismutase. AB - Chlorite dismutase carries out the heme-catalyzed decomposition of ClO2- to Cl- and O2, an unusual transformation with biotechnological and bioremediative applications. The enzyme has been successfully overexpressed for the first time in highly functional form in Escherichia coli and its steady state kinetics studied. The purified enzyme is abundant (55 mg/L cell culture), highly active (approximately 4.7 x 10(3) micromol of ClO2- min(-1) mg(-1) subunit) and nearly stoichiometric in heme; further, it shares spectroscopic and physicochemical features with chlorite dismutases previously isolated from three organisms. A careful study of the enzyme's steady state kinetics has been carried out. ClO2- consumption and O2 release rates were measured, yielding comparable values of kcat (4.5 x 10(5) min(-1)), K(m) (approximately 215 microM), and kcat/Km (3.5 x 10(7) M(-1) s(-1) via either method (4 degrees C, pH 6.8; all values referenced per heme-containing subunit). ClO2-:O2 stoichiometry exhibited a 1:1 relationship under all conditions measured. Though the value of kcat/Km indicates near diffusion control of the reaction, viscosogens had no effect on k(cat)/K(m) or V(max). The product O2 did not inhibit the reaction at saturating [O2], but Cl- is a mixed inhibitor with relatively high values of KI (225 mM for enzyme and 95.6 mM for the enzyme-substrate complex), indicating a relatively low affinity of the heme iron for halogen ions. Chlorite irreversibly inactivates the enzyme after approximately 1.7 x 10(4) turnovers (per heme) and with a half-life of 0.39 min, resulting in bleaching of the heme chromophore. The inactivation K(I) (K(inact)) of 166 microM is similar in magnitude to Km, consistent with a common Michaelis complex on the pathway to both reaction and inactivation. The one electron peroxidase substrate guaiacol offers incomplete protection of the enzyme from inactivation. Mechanisms in keeping with the available data and the properties of other well-described heme enzymes are proposed. PMID- 18422346 TI - Influence of multiple metal ions on beta-amyloid aggregation and dissociation on a solid surface. AB - Recently discovered evidences suggest that precipitation of Alzheimer's beta amyloid (Abeta) peptide and the toxicity in Alzheimer's disease (AD) are caused by abnormal interactions with neocortical metal ions, especially Zn2+, Cu2+, and Fe3+. While many studies had focused on the role of a "single" metal ion and its interaction with Abeta peptides, such studies involving "multiple" metal ions have hardly been explored. Here, to explore the nature of codeposition of different metals, two or more metal ions along with Abeta were incubated over a solid template prepared by immobilizing Abeta42 oligomers. The influence of Zn2+,Cu2+, and Fe3+ on Abeta aggregation was investigated by two approaches: co incubation and sequential addition. Our results using ex situ AFM, ThT-induced fluorescence, and FTIR spectroscopy indicated that the co-incubation of Cu2+, Zn2+, and Fe3+ significantly altered the morphology of aggregates. A concentration dependence study with mixed metal ions suggested that Zn2+ was required at much lower concentrations than Cu2+ to yield nonfibrillar amorphous Abeta deposits. In addition, sequential addition of Zn2+ or Cu2+ on fibrillar aggregates formed by Fe3+ demonstrated that Zn2+ and Cu2+ could possibly change the conformation of the aggregates induced by Fe3+. Our findings elucidate the coexistence of multiple metal ions through their interactions with Abeta peptides or its aggregates. PMID- 18422345 TI - Steady-state kinetics and mechanism of LpxD, the N-acyltransferase of lipid A biosynthesis. AB - LpxD catalyzes the third step of lipid A biosynthesis, the (R)-3-hydroxymyristoyl acyl carrier protein ( R-3-OHC14-ACP)-dependent N-acylation of UDP-3-O-[(R)-3 hydroxymyristoyl]-alpha-D-glucosamine [UDP-3-O-(R-3-OHC14)-GlcN]. We have now overexpressed and purified Escherichia coli LpxD to homogeneity. Steady-state kinetics suggest a compulsory ordered mechanism in which R-3-OHC14-ACP binds prior to UDP-3-O-(R-3-OHC14)-GlcN. The product, UDP-2,3-diacylglucosamine, dissociates prior to ACP; the latter is a competitive inhibitor against R-3-OHC14 ACP and a noncompetitive inhibitor against UDP-3-O-(R-3-OHC14)-GlcN. UDP-2-N-[(R) 3-Hydroxymyristoyl]-alpha-D-glucosamine, obtained by mild base hydrolysis of UDP 2,3-diacylglucosamine, is a noncompetitive inhibitor against both substrates. Synthetic (R)-3-hydroxylauroyl-methylphosphopantetheine is an uncompetitive inhibitor against R-3-OHC14-ACP and a competitive inhibitor against UDP-3-O-(R-3 OHC14)-GlcN, but (R)-3-hydroxylauroyl-methylphosphopantetheine is also a very poor substrate. A compulsory ordered mechanism is consistent with the fact that R 3-OHC14-ACP has a high binding affinity for free LpxD whereas UDP-3-O-(R-3-OHC14) GlcN does not. Divalent cations inhibit R-3-OHC14-ACP-dependent acylation but not (R)-3-hydroxylauroyl-methylphosphopantetheine-dependent acylation, indicating that the acidic recognition helix of R-3-OHC14-ACP contributes to binding. The F41A mutation increases the K(M) for UDP-3-O-(R-3-OHC14)-GlcN 30-fold, consistent with aromatic stacking of the corresponding F43 side chain against the uracil moiety of bound UDP-GlcNAc in the X-ray structure of Chlamydia trachomatis LpxD. Mutagenesis implicates E. coli H239 but excludes H276 as the catalytic base, and neither residue is likely to stabilize the oxyanion intermediate. PMID- 18422347 TI - The ability of actinic light to modify the bacteriorhodopsin photocycle revisited: heterogeneity vs photocooperativity. AB - In 1995, evidence both for photocooperativity and for heterogeneity as possible explanations for the ability of actinic light to modify the kinetics and pathways of the bacteriorhodopsin (BR) photocycle was reviewed ( Shrager, R. I., Hendler, R. W., and Bose, S. (1995) Eur. J. Biochem. 229, 589-595 ). Because both concepts could be successfully modeled to experimental data and there was suggestive published evidence for both, it was concluded that both photocooperativity and heterogeneity may be involved in the adaptation of the BR photocycle to different levels of actinic light. Since that time, more information has become available and it seemed appropriate to revisit the original question. In addition to the traditional models based on all intermediates in strict linear sequences, we have considered both homogeneous and heterogeneous models with branches. It is concluded that an explanation based on heterogeneity is more likely to be the true basis for the variation of the properties of the photocycle caused by changes in actinic light intensity. On the basis of new information presented here, it seems that a heterogeneous branched model is more likely than one with separate linear sequences. PMID- 18422348 TI - Predicting stability of DNA duplexes in solutions containing magnesium and monovalent cations. AB - Accurate predictions of DNA stability in physiological and enzyme buffers are important for the design of many biological and biochemical assays. We therefore investigated the effects of magnesium, potassium, sodium, Tris ions, and deoxynucleoside triphosphates on melting profiles of duplex DNA oligomers and collected large melting data sets. An empirical correction function was developed that predicts melting temperatures, transition enthalpies, entropies, and free energies in buffers containing magnesium and monovalent cations. The new correction function significantly improves the accuracy of predictions and accounts for ion concentration, G-C base pair content, and length of the oligonucleotides. The competitive effects of potassium and magnesium ions were characterized. If the concentration ratio of [Mg (2+)] (0.5)/[Mon (+)] is less than 0.22 M (-1/2), monovalent ions (K (+), Na (+)) are dominant. Effects of magnesium ions dominate and determine duplex stability at higher ratios. Typical reaction conditions for PCR and DNA sequencing (1.5-5 mM magnesium and 20-100 mM monovalent cations) fall within this range. Conditions were identified where monovalent and divalent cations compete and their stability effects are more complex. When duplexes denature, some of the Mg (2+) ions associated with the DNA are released. The number of released magnesium ions per phosphate charge is sequence dependent and decreases surprisingly with increasing oligonucleotide length. PMID- 18422349 TI - Electrogenic proton-pumping capabilities of the m-fast and m-slow photocycles of bacteriorhodopsin. AB - The parallel model for the bacteriorhodopsin (BR) photocycle at neutral pH and a temperature near 20 degrees C contains an M-fast cycle with steps BR-->K-->L-->Mf ->N-->O-->BR and an M-slow cycle which contains steps BR-->K-->L-->Ms-->BR. With increasing actinic laser strength, the M-fast cycle at first rises faster than the M-slow cycle, but reaches saturation sooner and at a lower level than the M slow cycle. The O-intermediate shows the same saturation behavior as Mf. In this paper, we show that the peak current of proton flux and the apparent voltages developed by this flux show the same saturation behavior as Ms, which is very different from that of both M f and O. It is further shown that most of the proton-charge displacement is connected with the step Ms-->BR. The optical and electrical data in these studies were collected simultaneously by a newly designed and built spectrometer which is described separately. PMID- 18422351 TI - Adsorption characteristics of bottle-brush polymers on silica: effect of side chain and charge density. AB - The adsorption behavior of bottle-brush polymers with different charge/PEO ratio on silica was studied using optical reflectometry and QCM-D. The results obtained under different solution conditions clearly demonstrate the existence of two distinct adsorption mechanisms depending on the ratio of charge/PEO. In the case of low-charge density brush polymers (0-10 mol %), the adsorption occurs predominantly through the PEO side chains. However, the presence of a small amount of charge along the backbone (as low as 2 mol %) increases the adsorption significantly above that of the uncharged bottle-brush polymer in pure water. As the charge density of the brush polymers is increased to 25 mol % or larger the adsorption occurs predominantly through electrostatic interactions. The adsorbed layer structure was studied by measuring the layer dissipation using QCM-D. The adsorbed layer formed by the uncharged brush polymer dissipates only a small amount of energy that indicates that the brush lie along the surface, the scenario in which the maximum number of PEO side chains interact with the surface. The adsorbed layers formed by the low-charge density brush polymers (2 10 mol %) in water are more extended, which results in large energy dissipation, whereas those formed by the high-charge density brush polymers (50-100 mol %) have their backbone relatively flat on the surface and the energy dissipation is again low. PMID- 18422350 TI - Pristine (C60) and hydroxylated [C60(OH)24] fullerene phototoxicity towards HaCaT keratinocytes: type I vs type II mechanisms. AB - The increasing use of fullerene nanomaterials has prompted widespread concern over their biological effects. Herein, we have studied the phototoxicity of gamma cyclodextrin bicapped pristine C 60 [(gamma-CyD) 2/C 60] and its water-soluble derivative C 60(OH) 24 toward human keratinocytes. Our results demonstrated that irradiation of (gamma-CyD) 2/C 60 or C 60(OH) 24 in D 2O generated singlet oxygen with quantum yields of 0.76 and 0.08, respectively. Irradiation (>400 nm) of C 60(OH) 24 generated superoxide as detected by the EPR spin trapping technique; superoxide generation was enhanced by addition of the electron donor nicotinamide adenine dinucleotide (reduced) (NADH). During the irradiation of (gamma-CyD) 2/C 60, superoxide was generated only in the presence of NADH. Cell viability measurements demonstrated that (gamma-CyD) 2/C 60 was about 60 times more phototoxic to human keratinocytes than C 60(OH) 24. UVA irradiation of human keratinocytes in the presence of (gamma-CyD) 2/C 60 resulted in a significant rise in intracellular protein-derived peroxides, suggesting a type II mechanism for phototoxicity. UVA irradiation of human keratinocytes in the presence of C 60(OH) 24 produced diffuse intracellular fluorescence when the hydrogen peroxide probe Peroxyfluor-1 was present, suggesting a type I mechanism. Our results clearly show that the phototoxicity induced by (gamma-CyD) 2/C 60 is mainly mediated by singlet oxygen with a minor contribution from superoxide, while C 60(OH) 24 phototoxicity is mainly due to superoxide. PMID- 18422352 TI - Dispersions of internally liquid crystalline systems stabilized by charged disklike particles as pickering emulsions: basic properties and time-resolved behavior. AB - The present paper reports on dispersions of internally liquid crystalline particles, formed from monoglyceride and oil mixtures, stabilized with discrete disklike particles of Laponite clay. Small-angle X-ray scattering (SAXS) was used to probe the presence of dispersed particles as well as their internal liquid crystalline structure. The data were compared to scattering results of reference systems, namely, from the bulk as well as from well-defined particles formed with a polymer as the emulsifier. The submicrometer sizes of the various particles could be derived using dynamic light scattering (DLS). The possible mechanisms involved in the stabilization of each of the different phases by the Laponite platelets, including the role of the residual salt, are discussed. Time-resolved experiments were performed over 60 days in order to follow the evolution of both the internal structure and size of the particles. In particular, we discuss the peculiar behavior of the sample without added oil, where the cubosomes transform into hexosomes over time. The effect of the high pH induced by the Laponite platelets in water, which could result in a hydrolysis of the monoglycerides, was shown to be responsible for the observed cubosome-to-hexasome transition, as well as for the decrease in the lattice parameters. PMID- 18422354 TI - Studying O-linked protein glycosylations in human plasma. AB - Recent investigations have implicated aberrant glycosylations in various malignancies, including epithelial ovarian cancer (EOC). The protocol here identifies O-linked carbohydrate patterns in EOC plasma glycoproteins through chemical cleavage and purification of these glycans. Dialyzed plasma is subjected to reductive beta-elimination with alkaline borohydride to release O-linked oligosaccharides from glycoproteins. Enrichment of released glycans, as well as removal of peptide and other contaminants, is followed by carbohydrate pattern analysis with MALDI-FT-ICR-MS. PMID- 18422353 TI - Linear discriminant analysis-based estimation of the false discovery rate for phosphopeptide identifications. AB - The development of liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) has made it possible to characterize phosphopeptides in an increasingly large-scale and high-throughput fashion. However, extracting confident phosphopeptide identifications from the resulting large data sets in a similar high-throughput fashion remains difficult, as does rigorously estimating the false discovery rate (FDR) of a set of phosphopeptide identifications. This article describes a data analysis pipeline designed to address these issues. The first step is to reanalyze phosphopeptide identifications that contain ambiguous assignments for the incorporated phosphate(s) to determine the most likely arrangement of the phosphate(s). The next step is to employ an expectation maximization algorithm to estimate the joint distribution of the peptide scores. A linear discriminant analysis is then performed to determine how to optimally combine peptide scores (in this case, from SEQUEST) into a discriminant score that possesses the maximum discriminating power. Based on this discriminant score, the p- and q-values for each phosphopeptide identification are calculated, and the phosphopeptide identification FDR is then estimated. This data analysis approach was applied to data from a study of irradiated human skin fibroblasts to provide a robust estimate of FDR for phosphopeptides. The Phosphopeptide FDR Estimator software is freely available for download at http://ncrr.pnl.gov/software/. PMID- 18422356 TI - Underdamped vibrations control the primary electron transfer in photosynthesis at low temperatures. AB - Measurements performed with the help of ultrafast laser spectroscopy have clearly shown that the primary electron transfer in photosynthesis lasts no longer than a few picoseconds. Equally fast are processes of vibrational relaxation which have to be taken into account in the correct description of the phenomenon. Here, we consider a simple theory combining the electron transfer process with diffusion in the energy space of a chosen underdamped vibrational mode of protein environment. Analytical formulas for effective transition rate constants are derived, and a transient kinetics is considered. The quality of analytical approximations is verified by numerical simulations for various physical conditions. Efficient parallel computations have been applied. The model can explain a peculiar temperature dependence of pump-probe spectra observed. PMID- 18422355 TI - Development and validation of sandwich ELISA microarrays with minimal assay interference. AB - Sandwich enzyme-linked immunosorbent assay (ELISA) microarrays are emerging as a strong candidate platform for multiplex biomarker analysis because of the ELISA's ability to quantitatively measure rare proteins in complex biological fluids. Advantages of this platform are high-throughput potential, assay sensitivity and stringency, and the similarity to the standard ELISA test, which facilitates assay transfer from a research setting to a clinical laboratory. However, a major concern with the multiplexing of ELISAs is maintaining high assay specificity. In this study, we systematically determine the amount of assay interference and noise contributed by individual components of a multiplexed 24-assay system. We find that nonspecific reagent cross-reactivity problems are relatively rare. We did identify the presence of contaminant antigens in a "purified antigen". We tested the validated ELISA microarray chip using paired serum samples that had been collected from four women at a 6-month interval. This analysis demonstrated that protein levels typically vary much more between individuals than within an individual over time, a result which suggests that longitudinal studies may be useful in controlling for biomarker variability across a population. Overall, this research demonstrates the importance of a stringent screening protocol and the value of optimizing the antibody and antigen concentrations when designing chips for ELISA microarrays. PMID- 18422357 TI - Solvent dependence of the photochemistry of the styrylpyridinium dye RH421. AB - RH421 is a widely used voltage-sensitive fluorescent membrane probe. It is also photochemically reactive and its photochemistry is dependent on its solvent surroundings. The aim of this study is to use the solvent dependence of the dye's photochemistry as an indicator of its environment within a lipid membrane. It was found that the dye actually undergoes two competing photochemical reactions. One photochemical pathway is characterized by an increase in fluorescence, the other by a fluorescence decrease. The pathway followed depends on the light intensity, the excitation wavelength and the solvent. The pathway leading to a fluorescence decrease is very dependent on the polarity of the solvent. It is promoted by solvents of high polarity (e.g., methanol, ethanol and propanol), by high intensity illumination and long wavelength excitation (i.e., 546 or 577 nm). In less polar solvents (butanol, pentanol, hexanol and heptanol) the pathway leading to a fluorescence increase dominates. The photochemical behavior observed when the dye is bound to a lipid membrane was not comparable to its behavior in any of the isotropic solvents studied. This is most likely due to the anisotropic environment of the membrane, which results in a steep gradient in polarity along the length of the dye molecule. PMID- 18422358 TI - Prediction of the solvation and structural properties of ionic liquids in water by two-dimensional correlation spectroscopy. AB - Two-dimensional vibrational spectroscopy is applied to investigate the dilution process of 1-ethyl-3-methyl-imidazolium tetrafluoroborate ([Emim][BF4]) in water. With increasing water content in ionic liquid (IL)/water mixtures, the C-H stretching vibration of the imidazolium cation showed systematic blue-shifts, which reflect the weakening of the cohesion between the cation and anion of ILs. The two-dimensional IR results reveal that the ILs sense quite different environments during the whole dilution process. First, the three-dimensional network structure of pure ILs was destroyed gradually into ionic clusters, then the clusters were further dissociated into ionic pairs surrounded by water molecules, and finally the latter became the dominant form in bulk water. Within the concentration range we investigated (0.02 or =40 years of age between 1994 and 2005. One control was matched to each PD case based on age, sex, general practice and index date. Lipid-lowering drug use was assessed by exposure timing (current vs past use) and by exposure duration (1-9, 10-29 or > or =30 prescriptions) prior to the index date for both cases and controls. Odds ratios (OR) were calculated using conditional logistic regression, adjusted for body mass index, smoking and various cardiovascular, metabolic and psychiatric co-morbidities. RESULTS: We identified 3637 cases with an incident idiopathic PD diagnosis, and the same number of controls. Compared with patients without hyperlipidaemia, those with untreated hyperlipidaemia did not have an altered relative PD risk (adjusted OR 0.98, 95% CI 0.74, 1.30). The adjusted ORs for current use of > or =30 prescriptions for statins or fibrates compared with non-use of statins or fibrates were 1.06 (95% CI 0.75, 1.51) and 1.25 (95% CI 0.51, 3.06), respectively. CONCLUSIONS: In this observational study, the long-term use of statins or fibrates was not associated with a substantially altered relative risk of developing PD. PMID- 18422381 TI - Inhaled corticosteroids and the risk of fractures in older adults: a systematic review and meta-analysis. AB - BACKGROUND: Inhaled corticosteroids (ICS) are commonly prescribed medications for the management of asthma and chronic obstructive pulmonary disease. It is well established that long-term use of these drugs may lower bone mineral density. However, whether ICS increase the risk of fractures remains unknown. Recent studies that have attempted to explore this risk have had conflicting results. We sought to explore the risk of ICS and fractures among older adults by conducting a systematic review and meta-analysis of the literature. METHODS: We systematically searched several databases, including MEDLINE, EMBASE and the Cochrane Library, to identify pertinent studies. Those studies that potentially met our inclusion criteria were identified by two reviewers. Relative risks (RRs) were pooled using the random effects model. We also explored dose-response by stratifying the analysis on high and low doses of ICS. Heterogeneity was assessed using the Q statistic and publication bias was assessed using the funnel plot. RESULTS: Thirteen studies, including four randomized controlled trials, were included in the review. The pooled RRs for hip fractures and any fractures were 0.91 (95% CI 0.87, 0.96) and 1.02 (95% CI 0.96, 1.08), respectively. When we restricted the analysis to users of high-dose ICS, the pooled RRs for any fractures and hip fractures were 1.30 (95% CI 1.07, 1.58) and 1.32 (95% CI 0.90, 1.92), respectively. The funnel plot did not show evidence of publication bias. CONCLUSION: We found no association between the use of ICS and fractures in older adults. A slight increase in risk was seen in those using high-dose ICS. The significance of this association should be investigated further. PMID- 18422382 TI - The European Herbal Medicines Directive: could it have saved the lives of Romeo and Juliet? AB - Herbal medicines have a long tradition of therapeutic use. However, they may occasionally cause dose-related (type A) or idiosyncratic (type B) toxicity and herb-drug interactions are also possible. Toxicity can arise as a result of misidentification or adulteration of the preparation. Legislation (the Directive on traditional herbal medicinal products 2004/24/EC) was enacted on 30 April 2004 to improve public health protection and promote the free movement of traditional medicinal products in the EU. It requires each Member State to set up a simplified registration scheme for manufactured traditional herbal medicines that are suitable for use without medical supervision. Evidence of 30 years of traditional use, at least 15 years of which should normally be within the EU, is required to permit minor claims, replacing the requirement to demonstrate efficacy. Safety is based on evidence in the published literature, although the regulator can also ask for more data if there are safety concerns. The pharmacovigilance requirements and quality standards are the same as for licensed medicines. Patient information is similar to that for any over-the-counter medicine, with an additional requirement for a statement on labels and in advertisements that the indication is based on traditional use. A European positive list of herbal substances will set out the indication, strength, dosing recommendations, route of administration and other information on safe use. Where a product complies with the list, the applicant will not need to demonstrate either the traditional use or the safety of the product. The list will be compiled by the recently established Committee on Herbal Medicinal Products at the European Medicines Agency. EU Member States were required to comply with the Directive by 30 October 2005. Traditional herbal medicinal products already on the market when the Directive became law need not comply with its provisions for 7 years after its coming into force. The public need to be aware that 'natural' does not necessarily mean 'safe' in all circumstances. They should be fully informed about all medicines they take. Consideration also needs to be given to effective regulation of herbal medicines practitioners, so that they are identifiable in law, are governed by professional codes of practice and have agreed standards of training and competency. There are many references to herbal medicines in Shakespeare's tragedy, Romeo and Juliet, which was written around 1595. A herbal medicine (distilled liquor) was almost certainly used to put Juliet into a deep sleep. A poison, possibly of herbal origin, was used by Romeo to take his own life when he thought his beloved Juliet was dead, rather than sleeping. While European herbal medicines regulation seeks to protect the public health by ensuring the necessary guarantees of quality, safety and efficacy, it was poor communication that appears to have triggered the chain of events leading to the death of Romeo and Juliet. Good communication between regulators, practitioners, patients and the public is necessary so that those who choose to take herbal medicines can do so with acceptable safety. PMID- 18422383 TI - Surveillance of suspected adverse reactions to natural health products: the case of propolis. AB - Natural health products are promoted to the public as equally or more effective and less toxic than conventional drugs. However, some 'natural' medicines are known to have adverse effects. From April 2002 to August 2007, 18 suspected adverse reactions associated with propolis-containing products were reported to the national surveillance system of natural health products, coordinated by the Italian National Health Institute. Sixteen reports concerned allergic reactions (with dermatological or respiratory symptoms), while two concerned the digestive tract. Some of the reactions were serious: six patients were admitted to hospital or visited an emergency department and in two of these a life-threatening event was reported. In seven patients (four of whom were children), an allergic predisposition was indicated. Propolis, a resinous substance collected by honeybees from the buds of living plants, has been used for several purposes (dermatitis, laryngitis, oral ulcers) because of its wide range of suggested activities (antibacterial, antiviral, antifungal, anti-inflammatory, antioxidant and chemopreventive actions). However, propolis is also a potent sensitizer and should not be used in patients with an allergic predisposition, in particular an allergy to pollen. In Italy, products containing bee derivatives (bee pollen, royal jelly or propolis) are available to the public as food supplements. No label warning of possible adverse reactions is found on the packaging, although it is well known that atopic and asthmatic individuals may be at an increased risk of allergic reactions after using these products. The public and healthcare practitioners should be aware of the risk of allergic reactions to products derived from bees and a warning should be added to the packaging of these products. PMID- 18422384 TI - Phytotherapic compounds: the consumer-pharmacist relationship. AB - Pharmacists play an important role in providing information about natural products and in preventing risks related to these substances, particularly with respect to interactions with conventional drugs. For these reasons, a survey was specifically designed to investigate the quality of self-care counselling by pharmacists on phytotherapy. Twenty-three pharmacy stores took part in the project. Face-to-face interviews, using a pre-structured questionnaire, were undertaken by trained pharmacists to consumers buying a herbal product. The questionnaire included socio-demographic data and 17 items designed to elicit information regarding the reason of consumption, product knowledge, relationship/communication with healthcare providers, level of satisfaction, concurrent drug use and adverse reactions. The collection of interviews started in November 2006 until April 2007. From the analysis of 1420 questionnaires, it is evident that herbal use is increasing in Italy: 12% of our interviewees were buying a herbal product for the first time. The present survey highlights the favourable perception of efficacy of phytotherapic compounds by the pharmacy's consumers, who consider this healthcare modality to be an important and effective way to promote health/wellness and disease management as well as being safer overall than conventional drugs. Moreover, findings from this study demonstrate that pharmacists are more likely to answer correctly about the uses of herbal medicines than about drug interactions, adverse drug effects and cautions about these products. PMID- 18422385 TI - European legislation on herbal medicines: a look into the future. AB - Harmonization of the market for herbal medicines is a fundamental requirement for European industries and health professionals and it will also be useful for consumers. Herbal medicines are generally sold as food supplements, but a common regulatory status in the various European countries does not exist. As a consequence, information on clinical indications for use, efficacy and safety are influenced by different opinions, according to the clinical or traditional experience of various folk medicines available in each European country. The European Directive 2004/24/EC released in 2004 by the European Parliament and by the Council of Europe provides the basis for the use of herbal medicines in Europe going forward. The Directive establishes that herbal medicines released in the market need authorization by the national regulatory authorities of each European country and that these products must have a recognized level of safety and efficacy. The safety of herbal medicinal products will be evaluated on the basis of existing scientific literature (data from clinical studies, case reports, pre-clinical studies). When data on safety are not sufficient, it will be communicated to consumers. According to the criteria of safety and efficacy, we will have two kinds of herbal medicinal products in the future: (i) 'well established use herbal medicinal products' (medicinal herbs with a recognized level of safety and efficacy); and (ii) 'traditional use herbal medicinal products'. The later category will include those medicinal herbs that do not have a recognized level of efficacy but are acceptably safe. Even though the fundamental objective of the new European herbal legislation is the harmonization of the market of herbal medicines, important regulations have been introduced, which will contribute to safer use of herbal substances if adopted by the whole of the European community. PMID- 18422386 TI - Understanding the adverse effects of cosmetics: a pilot project in cosmetovigilance. AB - Currently, cosmetics and toiletries are very popular and their use continues to increase because consumers consider physical appearance important and, at the same time, these products are considered to be safe. However, in spite of their safety and tolerability, during recent decades, we have become aware that adverse effects can occur. The number of adverse effects known so far is very low indeed. This is partly because such adverse effects are underestimated as a result of self-diagnosis and self-medication, which are common behaviours in the presence of mild-to-moderate reactions, as in the case of cosmetics. Moreover, such effects are underestimated because of the absence of formal and reliable monitoring systems ('cosmetovigilance'). This requires the creation of a standard reporting form, as well as resolution concerning professional categories authorized to report and the subsequent validation/evaluation of the collected forms. All these items are of great importance, not only to investigate but also to prevent risks caused by cosmetic use. A pilot project was undertaken to test the effectiveness of a notification system by the validation of either a reporting form or the role of dermatologists and community pharmacists as reporting categories. Collection of reporting forms began in July 2006 and it is still in progress; the preliminary data reported here refer to the period July 2006-June 2007 and mainly concern the recording and validation of the collected reporting forms. During this period, we have received 40 reporting forms (32 by dermatologists and 8 by pharmacists). The validation process of the recorded forms revealed several drawbacks, such as incompleteness (19 forms), inadequacy of the description of the suspected undesirable effect and its location (2), illegible handwriting (6) and mistaken statements (3). Six forms reported a misuse of a cosmetic product: four of these were related to the site of application while two were related to time. In one case, instructions for use were not followed. In conclusion, our experience regarding the notification of adverse effects of cosmetics, although limited to a restricted geographical area, suggests that for an efficient and reliable monitoring system to be in place, which includes all the necessary measures to protect public health, an education and training programme for all stakeholders (health professionals, consumers and appropriate authorities) is required. PMID- 18422387 TI - Cosmetovigilance: the 'beautiful' risk. AB - A cosmetic is classically defined as any preparation that is applied to the skin, eyes, mouth, hair or nails for the purpose of cleansing, enhancing appearance, giving a pleasant smell or giving protection. Unlike drugs, which are used to treat or prevent a disease in the body, cosmetics are not thought to change or affect the body's structure or functions. However, the distinction between drugs and cosmetics is sometimes not clear. Regulations for cosmetic products primarily address the safety of products that may be used by large populations of healthy consumers. However, the efficacy and safety of cosmetic products are not reviewed or approved by national authorities before they are sold to the public. The identification and analysis of adverse effects related to cosmetic products is a process that is currently still, to a large extent, industry driven. It is the responsibility of manufacturers to determine that products and ingredients are safe before they are marketed, and then to collect reports of adverse reactions. However, although the manufacturers do their best to monitor the safety profile of their products, we should consider that there is always a potential inherent conflict of interest. PMID- 18422388 TI - Assessment of the risk of fragrance allergy in the general population: challenges and methodological issues. AB - There are still unanswered questions about the safety of fragrances. In this conference paper, fragrance allergy will be considered in the context of a wider discussion concerning the prevalence and causes of contact dermatitis. No criteria for a reliable diagnosis of 'contact dermatitis' are available. International recommendations and standardization for the patch test method exist; however, the question of whether agents that are positive are causally linked to contact dermatitis remains fraught with uncertainties concerning false positive rates and clinical relevance. Most of the discussion concerning prevalence or incidence variations of allergic contact dermatitis to fragrances concentrate on the frequency of positive patch tests in clinical series, i.e. 'floating numerators'. Risk assessment requires that data from different sources are integrated and compared. Therefore, both a 'sentinel surveillance' system and more refined epidemiological studies in well defined populations are needed to reliably assess risks associated with fragrance exposure. PMID- 18422389 TI - TNF-alpha inhibitors and leukaemia: international pharmacovigilance reports. PMID- 18422390 TI - Long-term safety and efficacy of a combination of niacin extended release and simvastatin in patients with dyslipidemia: the OCEANS study. AB - INTRODUCTION: High-dose HMG-CoA reductase inhibitors (statins) fail to prevent approximately two-thirds of cardiovascular events. This fact has focused increased attention on treating abnormalities of non-high-density lipoprotein cholesterol (non-HDL-C), HDL-C, and triglycerides in national guidelines and has intensified interest in combination therapy. METHODS: The OCEANS study (Open label evaluation of the safety and efficacy of a Combination of niacin ER and simvAstatin in patieNts with dySlipidemia; ClinicalTrials.gov identifier: NCT00080275) evaluated the safety and efficacy of a combination of niacin extended release and simvastatin (NER/S; SIMCOR) over 52 weeks in 520 patients with mixed dyslipidemia. After a >or=4-week run-in phase of diet modification and simvastatin 40 mg/day, median baseline values (mg/dL) were: non-HDL-C = 141, low density lipoprotein-cholesterol (LDL-C) = 110, HDL-C = 45, and triglyceride = 151. Patients were randomized to an 8- or 12-week niacin titration scheme to a maximum NER/S dosage of 2,000/40 mg/day. RESULTS: Differences between titration groups in tolerability, safety, and efficacy were minimal; therefore, all results are for pooled titration groups. The safety of NER/S was consistent with the safety profile of each individual component. Treatment with NER/S was well tolerated: 71% of patients experienced flushing and 92% of flushing episodes were mild or moderate in intensity. Overall, 61% of patients experienced flushing episodes that were rated as mild or moderate in intensity. Flushing decreased over time: <40% of those who had flushing during titration experienced flushing during the final 12 weeks. A total of 20% of patients discontinued treatment because of a treatment-related adverse event, including 7% who discontinued because of flushing. Median changes from baseline (following the simvastatin 40 mg/day run-in phase) to 24 weeks were: non-HDL-C = -27.3%, LDL-C = -25.0%, HDL-C = +23.9%, and triglycerides = -35.9% (all p < 0.0001 vs baseline). In lipid treatment-naive patients, NER/S 2,000/40 mg/day decreased non-HDL-C, LDL-C, and triglycerides by approximately 50% and increased HDL-C by approximately 25% when week-24 lipid values were compared with lipid values obtained prior to the simvastatin 40 mg/day run-in. All three therapeutic lipid targets (LDL-C [risk adjusted goal], HDL-C >or=40 mg/dL, and triglycerides <150 mg/dL) were achieved concurrently by 65% of patients treated with NER/S. CONCLUSION: Treatment with NER/S 2,000/40 mg/day is well tolerated, has no unanticipated adverse events, and provides additional, clinically relevant improvements in multiple lipid parameters beyond statin monotherapy. PMID- 18422391 TI - Comparison of efficacy of low- (80 mg/day) and high- (160-320 mg/day) dose valsartan in the prevention of in-stent restenosis after implantation of bare metal stents in type B2/C coronary artery lesions. AB - BACKGROUND AND OBJECTIVE: Results from the VALVACE (VALsartan Versus ACE inhibition after bare metal stent implantation) trial suggest that prevention of in-stent restenosis after implantation of bare-metal stents in type B2/C coronary artery lesions is possible after administration of valsartan 80 mg/day. However, the restenosis rate in patients with stable angina was relatively high (27%) with this dosage and no different from patients taking ACE inhibitors. Therefore, a 1 : 1 matched comparison on a case-control basis was initiated in a prospective controlled registry using a higher dose of valsartan, 160-320 mg/day. METHODS: A total of 450 patients (241 men, mean age 62.7 +/- 9.1 years) with matched demographic and angiographic characteristics to patients in the VALVACE trial were treated with high-dose oral valsartan 160-320 mg/day over 6 months until control angiography. Angiographic restenosis rate, target lesion revascularization (TLR) and target vessel revascularization (TVR) rates, major adverse cardiac event (MACE) rate (death, myocardial infarction, and stent thrombosis) and mean late lumen loss were analysed after 6 months. Results were compared with the results of the VALVACE trial. Analysis of the combined results of the current study together with the VALVACE trial data enabled calculation of the gender- and dose-dependent effects of valsartan. RESULTS: In the high-dose valsartan group, the angiographic restenosis rate in 368 patients with control angiography was 7.3% compared with 19.5% in the low-dose group (VALVACE) [p < 0.0001]. Mean late lumen loss was 0.37 +/- 0.3 mm in the high-dose group compared with 0.53 +/- 0.31 mm in the VALVACE trial (p < 0.01). TLR and TVR rates were 4.3% in the high-dose group compared with 9% in the VALVACE trial (p < 0.01). The MACE rate was 0% in the high-dose group compared with 1.5% in the VALVACE trial (p < 0.01). Summarizing the data for valsartan, the in-stent restenosis rates in men were 22.7%, 13.3%, 6.7%, and 5.4% in patients receiving 80, 160, 240, and 320 mg/day, respectively. In women, the in-stent restenosis rates were 13.3% and 6.3% in patients receiving 80 and 160 mg/day, respectively; no restenosis occurred in patients receiving higher doses. CONCLUSION: Administration of high-dose oral valsartan 160-320 mg/day after implantation of bare-metal stent in type B2/C coronary artery lesions reduces angiographic in-stent restenosis, TLR, TVR, late lumen loss, and MACE rates more effectively than low-dose valsartan 80 mg/day. PMID- 18422392 TI - Which strategy is more effective for the treatment of cardiovascular disease: high-dose angiotensin II type 1 receptor antagonist monotherapy or combination therapy? AB - Angiotensin II type 1 (AT(1)) receptor antagonists (blockers) [ARBs] are highly selective for the AT(1) receptor and block the diverse effects of angiotensin II. When high BP is not controlled by low-dose ARB monotherapy, physicians need to employ another strategy, either high-dose ARB monotherapy or combination therapy with calcium channel antagonists (blockers) [CCBs], diuretics, or other agents. High-dose ARB monotherapy is more effective for decreasing proteinuria than low dose ARB monotherapy or CCBs. Although the ARB valsartan has been shown to prevent coronary restenosis in a clinical study (Val-PREST [Valsartan for prevention of restenosis after stenting of type B2/C lesions]), it is still unclear whether ARBs help to prevent restenosis. The results reported by Peters in this issue highlight the relative efficacies of low- (80 mg/day) and high-dose valsartan (160-320 mg/day) for the prevention of in-stent restenosis after the implantation of bare-metal stents, and suggest that high-dose valsartan can reduce the in-stent restenosis rate, target lesion revascularization and target vessel revascularization rates, late lumen loss, and major adverse cardiac events rate more effectively than low-dose valsartan. A better understanding of the differences in the efficacies of high- and low-dose ARBs could be useful in the treatment of patients with cardiovascular disease and may resolve the issue of whether ARBs prevent coronary restenosis. Clinical benefits may be induced by complete blockade of the renin-angiotensin system using high-dose ARB monotherapy. Therefore, physicians need to select a strategy carefully; i.e. either high-dose ARB monotherapy or combination therapy. PMID- 18422393 TI - Aspirin and platelet adenosine diphosphate receptor antagonists in acute coronary syndromes and percutaneous coronary intervention: role in therapy and strategies to overcome resistance. AB - Platelet activation and aggregation are key components in the cascade of events causing thrombosis following plaque rupture. Antiplatelet therapy is essential in the treatment of patients with acute coronary syndromes (ACS) and for those requiring percutaneous coronary intervention (PCI). Aspirin (acetylsalicylic acid) is a well established antiplatelet therapy and is mandated for secondary prevention of cardiovascular events following ACS. In patients with ACS, the addition of clopidogrel to aspirin is more effective than aspirin alone. For patients undergoing PCI, dual antiplatelet therapy with aspirin and clopidogrel is warranted. Aspirin should be continued indefinitely after PCI. Pretreatment of patients with clopidogrel prior to PCI lowers the incidence of cardiovascular events, yet the optimum timing of drug administration and dose are still being investigated, as is the duration of therapy following PCI. Late-stent thrombosis with drug-eluting stents has pushed the recommendation for duration of clopidogrel therapy up to 1 year and perhaps beyond, in patients without risks for bleeding. The concepts of aspirin and clopidogrel resistance are important clinical questions. No uniform definition exists for aspirin or clopidogrel resistance. Measurements of resistance are often highly variable and do not necessarily correlate with clinical resistance. Noncompliance remains the most prominent mode of resistance. Screening of selected patient populations for resistance or pharmacologic intervention of those patients termed 'resistant' warrants further study. PMID- 18422394 TI - Fondaparinux sodium: a review of its use in the management of acute coronary syndromes. AB - Fondaparinux sodium (Arixtra) is a synthetic, sulfated pentasaccharide, selective factor Xa inhibitor that is indicated in Europe for preventing thrombus formation in patients with acute coronary syndromes (ACS; the focus of this review), including those with ST-segment elevation myocardial infarction (STEMI), non STEMI (NSTEMI), or unstable angina.The large (n = 20,078), well designed OASIS-5 trial showed that subcutaneous fondaparinux 2.5 mg/day for 12,000) OASIS-6 trial. There were no differences in the incidence of major bleeding between these groups, resulting in a benefit : risk balance favoring fondaparinux. The specificity and selectivity of fondaparinux, combined with its long half-life and 100% bioavailability, allows once-daily anticoagulation without the need for monitoring activated clotting time. Subcutaneous fondaparinux was noninferior to enoxaparin treatment in patients with unstable angina or NSTEMI, and was more effective than usual care in those with STEMI. Fondaparinux has a favorable tolerability profile, particularly with regard to the risk of major bleeding, and limited data suggest that it is more cost effective than enoxaparin in the short term. Thus, overall, clinical evidence suggests that fondaparinux has a valuable place in the treatment of patients with ACS. PMID- 18422395 TI - Rosuvastatin: a review of its effect on atherosclerosis. AB - The HMG-CoA reductase inhibitor (statin) rosuvastatin (Crestor) is widely available for use in the management of dyslipidemia, and was recently approved in the US to slow the progression of atherosclerosis as part of a strategy to lower low-density lipoprotein-cholesterol (LDL-C) and total cholesterol (TC) to target levels. Rosuvastatin has greater lipid-lowering efficacy than any of the other currently available statins, and significantly more patients receiving rosuvastatin than other statins achieve LDL-C goals. Rosuvastatin delayed the progression of carotid atherosclerosis in patients with subclinical carotid atherosclerosis, moderately elevated cholesterol levels, and a low risk of cardiovascular disease in a primary prevention trial (METEOR). The results of METEOR suggest a possible role for the earlier use of rosuvastatin in primary prevention, although more data are needed from trials examining the effects of the drug on cardiovascular endpoints. Significant regression of atherosclerosis was seen with rosuvastatin 40 mg/day in patients with established coronary heart disease (CHD) in the ASTEROID trial, supporting the use of intensive lipid lowering in secondary prevention patients (although it should be noted that it has not yet been established that atherosclerotic regression translates into improved cardiovascular outcomes). Rosuvastatin is generally well tolerated, with a similar tolerability profile to that of other currently available statins. Thus, rosuvastatin is an important lipid-lowering treatment option that has been shown to cause regression of atherosclerosis in secondary prevention patients, and has a potential future role in delaying atherosclerosis in primary prevention patients. PMID- 18422396 TI - A multisite study of the capacity of acute stress disorder diagnosis to predict posttraumatic stress disorder. AB - OBJECTIVE: Previous studies investigating the relationship between acute stress disorder (ASD) and posttraumatic stress disorder (PTSD) have reported mixed findings and have been flawed by small sample sizes and single sites. This study addresses these limitations by conducting a large-scale and multisite study to evaluate the extent to which ASD predicts subsequent PTSD. METHOD: Between April 2004 and April 2005, patients admitted consecutively to 4 major trauma hospitals across Australia (N = 597) were randomly selected and assessed for ASD (DSM-IV criteria) during hospital admission (within 1 month of trauma exposure) and were subsequently reassessed for PTSD 3 months after the initial assessment (N = 507). RESULTS: Thirty-three patients (6%) met criteria for ASD, and 49 patients (10%) met criteria for PTSD at the 3-month follow-up assessment. Fifteen patients (45%) diagnosed with ASD and 34 patients (7%) not diagnosed with ASD subsequently met criteria for PTSD. The positive predictive power of PTSD criteria in the acute phase (0.60) was a better predictor of chronic PTSD than the positive predictive power of ASD (0.46). CONCLUSIONS: The majority of people who develop PTSD do not initially meet criteria for ASD. These data challenge the proposition that the ASD diagnosis is an adequate tool to predict chronic PTSD. PMID- 18422397 TI - Depressed patients with co-occurring alcohol use disorders: a unique patient population. AB - OBJECTIVE: Alcohol use and depressive disorders are frequently comorbid. Few studies have assessed the impact of comorbid alcohol use disorders (AUDs) on clinical aspects of major depression. We compared depressed subjects with and without co-occurring AUDs with respect to demographic and clinical parameters. METHOD: 505 individuals participated. 318 subjects had DSM-IV major depressive disorder (MDD) without a history of any alcohol or substance abuse/dependence (MDD only), and 187 individuals had MDD and a history of alcohol abuse/dependence (MDD/AUD). Demographic, clinical, and psychiatric history measures of patients in the 2 groups were examined and compared. The study was conducted from January 1990 to June 2005. RESULTS: MDD/AUD patients were younger at their first psychiatric hospitalization (p = .014), their first major depressive episode (p = .041), and their first suicide attempt (p = .001). They reported more previous major depressive episodes (p = .001), suicide attempts (p = .001), and recent life events (p = .001); and had higher lifetime aggression (p < .001), impulsivity (p < .001), and hostility (p < .001) scores. MDD/AUD patients were also more likely to report tobacco smoking (p < .001), a lifetime history of abuse (p = .004), and a history of AUD among first-degree relatives (p < .001) compared to MDD only patients. MDD/AUD individuals also had higher childhood (p < .001), adolescent (p < .001), and adult (p < .001) aggression scores and reported more behavioral problems during their childhood compared to their counterparts. Logistic regression analysis demonstrates that the number of previous depressive episodes, lifetime aggression, and smoking drive the difference between the groups. CONCLUSIONS: Our findings suggest that comorbid MDD/AUD may result from worse antecedents and lead to early onset, more comorbidity, and a more severe course of illness. PMID- 18422398 TI - Acute toxicity of thioarsenates to Vibrio fischeri. AB - Thioarsenic species often are the predominant arsenic species in sulfidic environments, yet little is known about their toxicity. We report to our knowledge the first determination of acute toxicity of mono-, di-, and trithioarsenate to the bioluminescent bacterium Vibrio fischeri, which increases with an increasing number of thio(SH)-groups. Whereas mono- and dithioarsenate are much less toxic (effective analyte concentration causing a 50% decrease in luminescence [EC50], 676 and 158 mg/L, respectively), the toxicity of trithioarsenate (EC50, 14.4 mg/L) is comparable to the toxicities of arsenate and arsenite (EC50, 9.1 and 26.1 mg/L, respectively). The low toxicity of monothioarsenate is remarkable, because it has chemical properties very similar to those of arsenate. In contrast to the toxicities of arsenite and arsenate, the toxicity of thioarsenates increases with exposure time, suggesting a lack of detoxification mechanisms or a conversion of thioarsenic species into arsenic oxyanions after uptake. We determined the acute toxicity of synthetic arsenite solutions with varying sulfide concentration to V. fischeri. Arsenic speciation in these solutions was measured by ion chromatography-inductively coupled plasma mass spectrometry, and the observed toxicity was related to the different arsenic species present. High inhibition of luminescence was observed at low and high ratios of sulfur to arsenic, in which arsenite or a mixture of di-, tri-, and tetrathioarsenate dominated arsenic speciation. Acute toxicity decreased at sulfur to arsenic ratios of from 1 to 10, with a minimum luminescence inhibition of 30% at a ratio of 3.5, at which concentrations of 55 mg/L of arsenite and 30 mg/L of trithioarsenate were determined. The toxicity observed under these conditions is much lower than that anticipated from the individual dose-response curves that predict each species alone already should cause 70 to 80% inhibition. The low toxicity suggests an antagonistic toxicological interaction between arsenite and trithioarsenate. PMID- 18422399 TI - Modeling the environmental fate of perfluorooctanoate and its precursors from global fluorotelomer acrylate polymer use. AB - The environment contains various direct and indirect sources of perfluorooctanoic acid (PFOA). The present study uses a dynamic multispecies environmental fate model to analyze the potential formation of perfluorooctanoate (PFO), the anion of PFOA, in the environment from fluorotelomer acrylate polymer (FTacrylate) emitted to landfills and wastewater, residual fluorotelomer alcohol (8:2 FTOH) in FTacrylate, and residual PFOA in FTacrylate. A multispecies version of the SimpleBox model, which is capable of determining the fate of a chemical and its degradation products, was developed for this purpose. An uncertainty analysis on the chemical-specific input parameters was performed to examine for uncertainty in modeled concentrations. In 2005, residual 8:2 FTOH made up 80% of the total contribution of FTacrylate use to PFO concentrations in global oceans, and residual PFOA in FTacrylate contributed 15% to PFO concentrations from FTacrylate use in global oceans. After hundreds of years, however, the main source of PFO from total historical FTacrylate production is predicted to be FTacrylate degrading in soil following land application of sludge from sewage treatment plants, followed by FTacrylate still present in landfills. Uncertainty in modeled PFO concentrations was up to a factor of 3.3. Current FTacrylate use contributes less than 1% of the PFO in seawater, but because direct PFOA emission sources are reduced and PFOA continues to be formed from FTacrylate in soil and in landfills, this fraction grows over time. PMID- 18422400 TI - Placental lesions and abnormal neurocognitive function at school age in extremely low-birth weight infants. PMID- 18422401 TI - Interexaminer agreement of histopathologic features in pediatric brain tumors: a biostatistical perspective. AB - Pages 108-117 of the current issue of Pediatric and Developmental Pathology contain a landmark interobserver investigation of some of the vagaries and vicissitudes involved in the nosological assessment of 6 histological features of childhood brain tumors. Doctor Gilles and his clinical research colleagues are to be commended for their conceptualization, reliability research design, execution, and interpretation of their findings. The piece is also uncommonly sophisticated concerning matters biostatistical. As such, it stands as a model for future nosological studies in childhood brain pathology, and can be applied more generally as well. The purpose of this Context article is to present some additional conceptual and methodologic issues that are relevant to the future design of histopathological studies in particular and of biobehavioral and biomedical diagnostic studies in general. In order to accomplish this goal, I shall discuss interexaminer reliability issues from the following viewpoints: type of reliability assessment, scales of measurement, reliability statistics, diagnostic accuracy, and the need to always differentiate levels of statistical significance from those that are both statistically and clinically meaningful, as Gilles and colleagues have also successfully accomplished. PMID- 18422402 TI - Generation of motor neurons by coculture of retinoic acid-pretreated embryonic stem cells with chicken notochords. AB - Understanding neuroectoderm formation and its subsequent diversification to functional neural subtypes remains elusive. We have shown here for the first time that embryonic stem cells (ESCs) can differentiate into neurons and motor neurons (MNs) by using a coculture embryonic notochord model in vitro. Mouse ESCs were induced to form neural precursors via timed exposure to retinoic acid (RA) using the 4-/4+ RA protocol. These cells were then cocultured with alginate bead encapsulated notochords isolated from Hamburger and Hamilton stage 6-10 chick embryos. The use of notochord alone was not able to induce neural differentiation from ESCs, and, therefore, notochord does not possess neural inducing activity. Hence, the most successful neuronal cells and MN differentiation was only observed following the coculture of RA-pretreated ESCs with notochord. This resulted in a significantly greater number of cells expressing microtubule associated protein-2 (MAP2), HB9, choline acetyltransferase (ChAT) and MN specific genes. While further characterization of these differentiated cells will be essential before transplantation studies commence, these data illustrate the effectiveness of embryonic notochord coculture in providing valuable molecular cues for directed differentiation of ESCs toward an MN lineage. PMID- 18422403 TI - Lighting in digital game worlds: effects on affect and play performance. AB - As a means of extending the significance of findings in experimental psychology and nonvisual psychological lighting research to digital game research, the present study was designed to investigate the impact of warm (reddish) and cool (bluish) simulated illumination in digital game worlds on game users' affect and play performance. In line with some previous findings, we predicted that lighting in a digital game world might, as in the real world, differently influence the nonvisual psychological mechanisms of affect, which in turn might enhance or impair the players' performance. It was shown that the players performed best and fastest in a game world lit with a warm (reddish) as compared to a cool (bluish) lighting. The former color of lighting also induced the highest level of pleasantness in game users. A regression analysis indicated tentatively that it was the level of pleasantness induced by the warm lighting that enhanced the players' better performance in that digital game world. It was also shown that high- as opposed to medium- or low-skilled players engage almost 2.5 times more per week in game playing. Given their skill, they performed significantly faster and felt significantly calmer and more relaxed in doing so. PMID- 18422404 TI - Antecedents of virtual community satisfaction and loyalty: an empirical test of competing theories. AB - The rapid growth of virtual communities (VCs) on the Internet raises important research questions regarding the antecedents of satisfaction and loyalty in VCs. This study compared the technology acceptance model (TAM), Seddon Information Systems Success Model (Seddon model), and an integrated model (combining the TAM and Seddon model) to examine which model best helps to predict user satisfaction with VCs. Using a structural equation model, data collected from 198 community members were used to compare the three models in terms of overall model fit, explanatory power, and path significance. The findings show that two TAM components (perceived usefulness and ease of use) are determinants of user satisfaction with VCs. This study also found the influence of quality-perception dimensions (e.g., system and information quality) on user satisfaction to be significant. Additionally, this study found general support for user satisfaction as a determinant of loyalty in VCs. Finally, an integrated model, combining the TAM and Seddon model, provided better explanatory power than either the TAM or the Seddon model alone. Finally, this study discusses the implications of these findings and offers directions for future research. PMID- 18422405 TI - Personality traits and life satisfaction among online game players. AB - The DFC Intelligence predicts worldwide online game revenues will reach $9.8 billion by 2009, making online gaming a mainstream recreational activity. Understanding online game player personality traits is therefore important. This study researches the relationship between personality traits and life satisfaction in online game players. Taipei, Taiwan, is the study location, with questionnaire surveys conducted in cyber cafe shops. Multiple regression analysis studies the causal relationship between personality traits and life satisfaction in online game players. The result shows that neuroticism has significant negative influence on life satisfaction. Both openness and conscientiousness have significant positive influence on life satisfaction. Finally, implications for leisure practice and further research are discussed. PMID- 18422406 TI - Students' perceptions of constructivist Internet learning environments by a physics virtual laboratory: the gap between ideal and reality and gender differences. AB - The purpose of this study was to investigate the perceptions of Internet users of a physics virtual laboratory, Demolab, in Taiwan. Learners' perceptions of Internet-based learning environments were explored and the role of gender was examined by using preferred and actual forms of a revised Constructivist Internet based Learning Environment Survey (CILES). The students expressed a clear gap between ideal and reality, and they showed higher preferences for many features of constructivist Internet-based learning environments than for features they had actually learned in Demolab. The results further suggested that male users prefer to be involved in the process of discussion and to show critical judgments. In addition, male users indicated they enjoyed the process of negotiation and discussion with others and were able to engage in reflective thoughts while learning in Demolab. In light of these findings, male users seemed to demonstrate better adaptability to the constructivist Internet-based learning approach than female users did. Although this study indicated certain differences between males and females in their responses to Internet-based learning environments, they also shared numerous similarities. A well-established constructivist Internet-based learning environment may encourage more female learners to participate in the science community. PMID- 18422407 TI - Do aggressive people play violent computer games in a more aggressive way? Individual difference and idiosyncratic game-playing experience. AB - ABSTRACT This study investigates whether individual difference influences idiosyncratic experience of game playing. In particular, we examine the relationship between the game player's physical-aggressive personality and the aggressiveness of the player's game playing in violence-oriented video games. Screen video stream of 40 individual participants' game playing was captured and content analyzed. Participants' physical aggression was measured before the game play. The results suggest that people with more physical-aggressive personality engage in a more aggressive style of playing, after controlling the differences of gender and previous gaming experience. Implications of these findings and direction for future studies are discussed. PMID- 18422408 TI - Early sexual experiences: the role of Internet access and sexually explicit material. AB - The current study investigated whether viewing X-rated movies, Internet access in the home, and gender of the participant would differ between age of first initiation for oral sex, age of first initiation for sexual intercourse, and number of sexual partners. An online sample of 437 participants with an average age of 29.46 participated in the study. Each participant completed a survey that assessed early sexual behaviors and Internet and X-rated material exposure. Results discovered males with Internet access during the ages of 12 to 17 reported significantly younger ages for first oral sex compared to males without Internet access. In addition, male and female participants with Internet access, between the ages 12 to 17, reported younger ages for first sexual intercourse compared to participants without Internet access. Study limitations and implications are discussed. PMID- 18422409 TI - MySpace and Facebook: applying the uses and gratifications theory to exploring friend-networking sites. AB - The increased use of the Internet as a new tool in communication has changed the way people interact. This fact is even more evident in the recent development and use of friend-networking sites. However, no research has evaluated these sites and their impact on college students. Therefore, the present study was conducted to evaluate: (a) why people use these friend-networking sites, (b) what the characteristics are of the typical college user, and (c) what uses and gratifications are met by using these sites. Results indicated that the vast majority of college students are using these friend-networking sites for a significant portion of their day for reasons such as making new friends and locating old friends. Additionally, both men and women of traditional college age are equally engaging in this form of online communication with this result holding true for nearly all ethnic groups. Finally, results showed that many uses and gratifications are met by users (e.g., "keeping in touch with friends"). Results are discussed in light of the impact that friend-networking sites have on communication and social needs of college students. PMID- 18422410 TI - Organizational culture, creative behavior, and information and communication technology (ICT) usage: a facet analysis. AB - Despite the prominence of organizational culture (OC), this concept is controversial and its structure has yet to be systematically analyzed. This study develops a three-pronged formal definitional framework on the basis of facet theory (FT) and explores behavior modality, referent, and object. This facet analysis (FA) of OC accounts successfully for variation in both creative behavior at work and the usage of information and communication technologies (ICTs). An analysis of data collected from 230 employees in the financial industry indicates that a radex structure was obtained for work and ICT. The behavior modality facet ordered the space from center to periphery, and referents facet relates to the direction angles away from the origin. PMID- 18422411 TI - Neural correlates of "absence" in interactive simulator protocols. AB - While much has been written about the importance of the experience of presence in simulation protocols, the role of "absence" deserves closer attention. The role of subjective experience and neurophysiologic processes involved in fluctuating states of consciousness is a key issue in developing assessment and treatment tools using interactive immersive simulator tasks. This paper proposes that when engaging in an interactive simulator task, there are fluctuations of consciousness that determine both motivational engagement and measured performance. Rather than expecting a continuous experience, both in terms of perceptual and motor output flow, factors such as circadian fluctuations, fatigue, and actual intrusion of sleep into waking consciousness are relevant in assessments and treatments using virtual environment-based tasks. These factors are particularly relevant in treatment populations with neurological and psychiatric disorders, where such disturbances are more common. To illustrate this construct, a series of experiments examining these phenomena in relation to a standardized driving-simulation protocol to detect psychomotor impairment developed for clinical and diagnostic testing in a sleep laboratory setting are reviewed. PMID- 18422412 TI - Lessons learned: the effect of prior technology use on Web-based interventions. AB - This study examined the role of regular prior technology use in treatment response to an online family problem-solving (OFPS) intervention and an Internet resource intervention (IRI) for pediatric traumatic brain injury (TBI). Participants were 150 individuals in 40 families of children with TBI randomly assigned to OFPS intervention or an IRI. All families received free computers and Internet access to TBI resources. OFPS families received Web-based sessions and therapist-guided synchronous videoconferences focusing on problem solving, communication skills, and behavior management. All participants completed measures of depression, anxiety, and computer usage. OFPS participants rated treatment satisfaction, therapeutic alliance, and Web site and technology comfort. With the OFPS intervention, depression and anxiety improved significantly more among technology using parents (n = 14) than nontechnology users (n = 6). Technology users reported increasing comfort with technology over time, and this change was predictive of depression at followup. Satisfaction and ease-of-use ratings did not differ by technology usage. Lack of regular prior home computer usage and nonadherence were predictive of anxiety at followup. The IRI was not globally effective. However, controlling for prior depression, age, and technology at work, there was a significant effect of technology at home for depression. Families with technology experience at home (n = 11) reported significantly greater improvements in depression than families without prior technology experience at home (n = 8). Although Web-based OFPS was effective in improving caregiver functioning, individuals with limited computer experience may benefit less from an online intervention due to increased nonadherence. PMID- 18422413 TI - E-mentoring for youth with special needs: preliminary results. AB - Recently, a growing number of electronic mentoring (e-mentoring) Web sites have become available that provide vocational and career support. To date, few researchers have addressed the phenomenon of e-mentoring from a socioemotional perspective for populations with special needs. This paper presents a pilot study designed to test and evaluate an e-mentoring intervention program based on mutual self-disclosure and friendship for youth with special needs. Using qualitative methods, the study characterized the e-mentoring process and its contributions to this population. Results provided support for the socioemotional potential of computer-mediated communication for youth with special needs, although some barriers were found. Practical implications for implementing feasible e-mentoring programs for youth with special needs are discussed. PMID- 18422414 TI - Dynamic Interviewing Program (DIP): automatic online interviews via the instant messenger ICQ. AB - A Dynamic Interviewing Program (DIP) is presented for automated online interviewing via the instant messaging service ICQ. DIP can conduct parallel structured interviews with several ICQ users at a time, 24 hours a day. The interview itself can be dynamic (branched testing), which means that each question depends on the answer given to the previous one. An online ICQ address book offers the possibility to validate demographic data in order to evaluate data quality. Experiences with DIP and a first empirical study open up new perspectives for Internet-based automatic interviewing. PMID- 18422415 TI - Internet use and loneliness in older adults. AB - Use of the Internet by seniors as a communication technology may lead to changes in older adult social relationships. This study used an online questionnaire to survey 222 Australians over 55 years of age on Internet use. Respondents primarily used the Internet for communication, seeking information, and commercial purposes. The results showed negative correlations between loneliness and well-being. Multiple regression analyses revealed that greater use of the Internet as a communication tool was associated with a lower level of social loneliness. In contrast, greater use of the Internet to find new people was associated with a higher level of emotional loneliness. PMID- 18422416 TI - Induced attitude change on online gaming among adolescents: an application of the less-leads-to-more effect. AB - The negative impact of Internet use on adolescents has received much popular attention and has also become a popular research topic. How to induce adolescent players to change their attitudes toward online gaming is one of the most important issues in online gaming addiction. The present study is based on the less-leads-to-more effect of dissonance theory. Experimental research was conducted to examine the effects of rewards and decision freedom on attitude change toward online gaming among adolescents considered at risk for addiction. The results supported predictions based on external justification in dissonance theory. Specifically, fewer rewards produced greater attitude change toward online gaming in the condition of personal freedom of choice after participants exhibited attitude-discrepant behavior. However, the less-leads-to-more effect was not prominent in the condition without personal freedom of choice. Adopting a reward strategy to induce game players to disengage online gaming is discussed. PMID- 18422417 TI - Cyberbullying: youngsters' experiences and parental perception. AB - This paper describes a survey on the prevalence and nature of cyberbullying of and by youngsters and the parental perceptions. Two questionnaires, one for youngsters and one for their parents, were sent to 1,211 final-year pupils of primary schools and first-year pupils of all levels of secondary schools and their parents. Pupils completed the questionnaires in their classroom, and parents completed them at home. The results show that about 16% of the youngsters had engaged in bullying via the Internet and text messages, while about 23% of the youngsters had been victims of cyberbullying. Cyberbullying seems to be a rather anonymous, individualistic activity, which primarily takes place at home. The most frequently used nature of harassment were name-calling and gossiping. Youngsters mostly react to cyberbullying by pretending to ignore it, by really ignoring it, or by bullying the bully. Most parents set rules for their children about the way to they should use the Internet but are not really conscious of the harassments. They underestimate their own children's bullying behavior and have insufficient notion of their children as victims of bullying. PMID- 18422418 TI - Are internet cafes gendered spaces? AB - This study compared gender differences in the intention and behavior of university students with regard to visiting Internet cafes. The results revealed that males experienced significantly lower social pressure and had more positive attitudes than females. Males had significantly higher perceptions of control, past behavior, intention, and behavior than did females. Additionally, past behavior was the most influential factor in predicting intention and behavior. Two conclusions were drawn: (1) Internet cafes were speculated as a masculine gaming space and thus considered highly gendered. (2) Male and female respondents exhibited similar patterns when predicting their intention and behavior toward visiting Internet cafes. PMID- 18422419 TI - Sensation-seeking, Internet dependency, and online interpersonal deception. AB - This study aimed to elaborate the relationships between sensation-seeking, Internet dependency, and online interpersonal deception. Of the 707 individuals recruited to this study, 675 successfully completed the survey. The results showed high sensation-seekers and high Internet dependents were more likely to engage in online interpersonal deception than were their counterparts. PMID- 18422420 TI - Personality differences between online game players and nonplayers in a student sample. AB - This study examines the differences in openness, conscientiousness, and extraversion between online game players and nonplayers using a student sample. The matched sample comprised 130 pairs of students, who are paired in gender and age. The results indicate that online game players reported higher scores in openness, conscientiousness, and extraversion than did nonplayers. PMID- 18422421 TI - Exploring the personality trait of self-monitoring on technology usage of Web portals. AB - This study explores the personality trait of self-monitoring in Web portal usage by employing the extended theory of planned behavior (TPB) model of Pavlou and Fygenson. A survey of 254 Internet Web portal users was conducted to test the hypothesized model. The results show that our data supported all five individual causal paths postulated in this study. Further, the construct of self-monitoring plays a moderating role in Web portal usage; three of the five moderating hypotheses are supported. Based on the research findings, conclusions and implications are discussed. PMID- 18422422 TI - Gender differences in "social portraits" reflected in MySpace profiles. AB - The internet has brought about an entirely new method of self-presentation in such online social networking Web sites as MySpace in which individuals create profiles that reflect their identity. This cyber social tool provides a new site of analysis to examine the extent of patterns of gendered identity in which females tend to turn to others for validation in contrast to males, who are more apt to maintain their individuality and whose relationships are more of an extension of their already-complete selves. In this study of 51 female and 49 male MySpace profiles, males were less apt to mention their significant other in the "About Me" section: 43% mentioned their significant other 0 times compared to 16% of females, and 14% of males mentioned their significant other between 2 and 10 times compared to 37% of females (p = 0.003). In the "Interests" section, the majority of males (67%) did not mention their significant other at all compared to 47% of females, and 33% mentioned her between 1 and 5 times compared to 53% of females (p = 0.05). These results reveal that online data sources manifest identity formation consistent with traditional gender roles in which females are dependent on others for their sense of self. PMID- 18422427 TI - Yeast as a model host to explore plant virus-host interactions. AB - The yeast Saccharomyces cerevisiae is invaluable for understanding fundamental cellular processes and disease states of relevance to higher eukaryotes. Plant viruses are intracellular parasites that take advantage of resources of the host cell, and a simple eukaryotic cell, such as yeast, can provide all or most of the functions for successful plant virus replication. Thus, yeast has been used as a model to unravel the interactions of plant viruses with their hosts. Indeed, genome-wide and proteomics studies using yeast as a model host with bromoviruses and tombusviruses have facilitated the identification of replication-associated factors that affect host-virus interactions, virus pathology, virus evolution, and host range. Many of the host genes that affect the replication of the two viruses, which belong to two dissimilar virus families, are distinct, suggesting that plant viruses have developed different ways to utilize the resources of host cells. In addition, a surprisingly large number of yeast genes have been shown to affect RNA-RNA recombination in tombusviruses; this opens an opportunity to study the role of the host in virus evolution. The knowledge gained about host-virus interactions likely will lead to the development of new antiviral methods and applications in biotechnology and nanotechnology, as well as new insights into cellular functions of individual genes and the basic biology of the host cell. PMID- 18422428 TI - Living in two worlds: the plant and insect lifestyles of Xylella fastidiosa. AB - Diseases caused by Xylella fastidiosa have attained great importance worldwide as the pathogen and its insect vectors have been disseminated. Since this is the first plant pathogenic bacterium for which a complete genome sequence was determined, much progress has been made in understanding the process by which it spreads within the xylem vessels of susceptible plants as well as the traits that contribute to its acquisition and transmission by sharpshooter vectors. Although this pathogen shares many similarities with Xanthomonas species, such as its use of a small fatty acid signal molecule to coordinate virulence gene expression, the traits that it utilizes to cause disease and the manner in which they are regulated differ substantially from those of related plant pathogens. Its complex lifestyle as both a plant and insect colonist involves traits that are in conflict with these stages, thus apparently necessitating the use of a gene regulatory scheme that allows cells expressing different traits to co-occur in the plant. PMID- 18422426 TI - Role of stomata in plant innate immunity and foliar bacterial diseases. AB - Pathogen entry into host tissue is a critical first step in causing infection. For foliar bacterial plant pathogens, natural surface openings, such as stomata, are important entry sites. Historically, these surface openings have been considered as passive portals of entry for plant pathogenic bacteria. However, recent studies have shown that stomata can play an active role in limiting bacterial invasion as part of the plant innate immune system. As a counter defense, the plant pathogen Pseudomonas syringae pv. tomato DC3000 uses the virulence factor coronatine to actively open stomata. In nature, many foliar bacterial disease outbreaks require high humidity, rain, or storms, which could favor stomatal opening and/or bypass stomatal defense by creating wounds as alternative entry sites. Further studies on microbial and environmental regulation of stomatal closure and opening could fill gaps in our understanding of bacterial pathogenesis, disease epidemiology, and microbiology of the phyllosphere. PMID- 18422429 TI - Breaking the barriers: microbial effector molecules subvert plant immunity. AB - Adaptation to specialized environments allows microorganisms to inhabit an enormous variety of ecological niches. Growth inside plant tissues is a niche offering a constant nutrient supply, but to access this niche, plant defense mechanisms ranging from passive barriers to induced defense reactions have to be overcome. Pathogens have to break several, if not all, of these barriers. For this purpose, they secrete effector molecules into plant cells to interfere with individual defense responses. Plant defense is organized in multiple layers, and therefore the action of effectors likely follows this same order, leading to a hierarchy in effector orchestration. In this review we summarize the latest findings regarding the level at which effectors manipulate plant immunity. Particular attention is given to those effectors whose mechanism of action is known. Additionally, we compare methods to identify and characterize effector molecules. PMID- 18422431 TI - The continuing saga of MRSA. PMID- 18422432 TI - Hepatitis A: the vaccine dividend. PMID- 18422433 TI - Making universal access a reality--what more do we need to know? PMID- 18422434 TI - Changes in the prevalence of nasal colonization with Staphylococcus aureus in the United States, 2001-2004. AB - BACKGROUND: Staphylococcus aureus is a common cause of infection, particularly in persons colonized by this organism. Virulent strains of methicillin-resistant S. aureus (MRSA) have emerged in the general community. METHODS: A nationally representative survey of nasal colonization with S. aureus was conducted from 2001 through 2004 as part of the National Health and Nutrition Examination Survey. MRSA isolates were identified by the oxacillin disk-diffusion method. The pulsed-field gel electrophoresis (PFGE) type was determined for all MRSA isolates. A t statistic was used to compare the prevalence of colonization across biennia and across population subgroups. Cofactors independently associated with colonization were determined with backward stepwise logistic modeling. RESULTS: The prevalence of colonization with S. aureus decreased from 32.4% in 2001-2002 to 28.6% in 2003-2004 (P < .01), whereas the prevalence of colonization with MRSA increased from 0.8% to 1.5% (P < .05). Colonization with MRSA was independently associated with healthcare exposure in males and with having been born in the United States, age > or =60 years, diabetes, and poverty in females. In 2003 2004, a total of 19.7% (95% confidence interval, 12.4%-28.8%) of MRSA-colonized persons carried a PFGE type associated with community transmission. CONCLUSIONS: Nasal colonization with MRSA has increased in the United States, despite an overall decrease in nasal colonization with S. aureus. PFGE types associated with community transmission only partially account for the increase in MRSA colonization. PMID- 18422435 TI - What is community-associated methicillin-resistant Staphylococcus aureus? AB - BACKGROUND: A community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) infection has been defined as an MRSA infection in a patient who lacks specific risk factors for healthcare exposure. We sought to determine whether the absence or presence of these risk factors still predicts the phenotypic or genotypic characteristics of MRSA strains. METHODS: All clinical MRSA isolates were prospectively collected at the University of Chicago Hospitals from July 2004 through June 2005. Patients were interviewed and/or their medical records were reviewed. Isolates underwent genotyping and susceptibility testing. Data on patients and isolates were stratified in accordance with 8 frequently cited criteria for the identification of CA-MRSA and compared for concordance. RESULTS: Among 616 unique patients from whom MRSA isolates were recovered, 404 (65.6%) had risk factors for healthcare exposure. Of the 404 isolates recovered from these patients, 166 (41.1%) were clindamycin susceptible, 190 (47.0%) carried staphylococcal cassette chromosome mec (SCCmec) type IV, 145 (35.9%) carried the Panton-Valentine leukocidin genes (PVL+), and 162 (40.1%) were identified as sequence type (ST) 8 by multilocus sequence typing (MLST), all of which are characteristics commonly attributed to CA-MRSA strains. CONCLUSIONS: Association with the healthcare environment now has little predictive value for distinguishing patients with infection due to multidrug resistant MRSA isolates from those infected by CA-MRSA isolates, that is, isolates that are clindamycin susceptible, PVL+, ST8, and/or contain SCCmec type IV. Defining CA-MRSA by the absence of risk factors for healthcare exposure greatly underestimates the burden of epidemic CA-MRSA disease. PMID- 18422436 TI - Host polymorphisms in interleukin 4, complement factor H, and C-reactive protein associated with nasal carriage of Staphylococcus aureus and occurrence of boils. AB - BACKGROUND: Staphylococcus aureus is capable of persistently colonizing the vestibulum nasi. We hypothesized that polymorphisms in host inflammatory response genes and genetic variation in S. aureus contribute to susceptibility to S. aureus carriage and infection. METHODS: The prevalence of persistent nasal carriage of S. aureus in 3851 participants aged 61-101 years was 18% (678 of 3851 participants), whereas 73% of volunteers (2804 of 3851) were not colonized. A total of 1270 individuals had boils. Polymorphisms in TNFA (C -863T), IL4 (C 542T), CFH (Tyr402His), and CRP (C1184T, C2042T, and C2911G) were determined. Genetic similarity among 428 S. aureus isolates was determined by use of amplified fragment length polymorphism analysis (AFLP)-mediated genotyping. RESULTS: The IL4 -524 C/C host genotype was associated with an increased risk of persistent S. aureus carriage, irrespective of S. aureus AFLP genotype. The CRP haplotype 1184C; 2042C; 2911C was overrepresented in individuals who were not colonized . In individuals with boils, carriers of the CFH Tyr402 variant, and the CRP 2911 C/C genotype were overrepresented. CONCLUSION: Persistent carriage of S. aureus is influenced by genetic variation in host inflammatory response genes. As would be expected in multifactorial host-microbe interactions, these effects are limited. Interestingly, host genotype was associated with the carriage of certain S. aureus genotypes. Apparently, a close interaction between host and bacterial determinants are prerequisites for long-term colonization. PMID- 18422437 TI - SarZ is a key regulator of biofilm formation and virulence in Staphylococcus epidermidis. AB - Biofilm-associated infection due to Staphylococcus epidermidis, the leading nosocomial pathogen, is a major problem for the public health system, but the regulation of this important phenotype is not completely understood. Using a highly discriminatory screening procedure for genes that influence biofilm formation, we identified the transcriptional regulator SarZ as a novel important determinant of biofilm formation and biofilm-associated infection, on the basis of the significant impact of sarZ on the transcription of the biosynthetic operon for biofilm exopolysaccharide. In addition, sarZ influenced the expression of a series of virulence genes, including genes that influence the expression of lipases and proteases, resistance to an important human antimicrobial peptide, and hemolysis. Our study indicates that the SarZ regulator has a key role in maintaining the typical S. epidermidis phenotype, which is characterized by pronounced biofilm formation and immune evasion, a likely reason for the success of S. epidermidis as a colonizing organism and pathogen in chronic, biofilm associated infection. PMID- 18422438 TI - Efficacy of recombinant bacille Calmette-Guerin vaccine secreting interleukin 15/antigen 85B fusion protein in providing protection against Mycobacterium tuberculosis. AB - Protection against Mycobacterium tuberculosis not only depends on CD4+ T helper type 1 (Th1) cells but, also, on CD8+ T cells. Interleukin (IL)-15 has an important function in the maintenance of memory CD8+ T cells. In the present study, we examined the efficacy of recombinant Mycobacterium bovis bacille Calmette-Guerin (rBCG) secreting fusion protein antigen (Ag) 85B murine IL-15 (rBCG-Ag85B-IL15) in providing protection against M. tuberculosis infection. The levels of major histocompatibility (MHC) class Ib (H2-M3)-binding TB2- or MHC class Ia (H-2Db)-binding MPT64-specific CD8+ T cells producing interferon (IFN) gamma were significantly higher after immunization with rBCG-Ag85B-IL15 than after immunization with rBCG secreting Ag85B (rBCG-Ag85B). The levels of purified protein derivative- or Ag85B-specific CD4+ T cells producing IFN-gamma were also higher in mice immunized with rBCG-Ag85B-IL15 than in mice immunized with rBCG Ag85B. Mice immunized with rBCG-Ag85B-IL15 exhibited CD8+ and CD4+ T cells responses that were stronger than those in mice immunized with rBCG-Ag85B, as well as robust protection in the lung against intratracheal challenge of M. tuberculosis. Thus, rBCG-Ag85B-IL15 vaccination capable of inducing efficient cell-mediated immunity might be used as an effective vaccine for tuberculosis. PMID- 18422439 TI - School-aged children: a reservoir for continued circulation of Haemophilus influenzae type b in the United Kingdom. AB - BACKGROUND: A resurgence of Haemophilus influenzae type b (Hib) disease occurred in the United Kingdom between 1999 and 2003 and was partially attributed to lower immunogenicity of combination vaccines. The reservoir for Hib that led to transmission in this period is unknown. METHODS: We estimated the point prevalence of Hib carriage in school-aged children and adults, using oropharyngeal swabbing and selective media. We characterized the Hib isolates by multilocus sequence typing (MLST) and measured Hib antibody concentrations in adults by enzyme-linked immunosorbent assay. RESULTS: Point prevalence for Hib carriage in 855 children aged 6-16 years was 4.2% (95% confidence interval [CI], 2.5%-5.9%). Five clonal groups of Hib were identified by MLST, 86% from the lineage of sequence type 6. No Hib was isolated in 385 adults (upper limit of 95% CI, 0.95%). The geometric mean concentration of serum antibody to polyribosylribitol phosphate was 0.47 microg/mL (95% CI, 0.37-0.59 mirog/mL) in adults. CONCLUSIONS: Hib carriage is common in school-aged children, who are a significant reservoir for ongoing transmission of Hib to susceptible individuals in the United Kingdom. Surveillance of transmission and immunity across all ages of the population is essential to monitor the evolution of Hib epidemiology. PMID- 18422440 TI - Declining hepatitis A mortality in the United States during the era of hepatitis A vaccination. AB - BACKGROUND: Since the mid-1990s, hepatitis A vaccine has been recommended for US children living in historically high-incidence states and for persons with other risk factors or chronic liver disease (CLD). The incidence of hepatitis A has declined dramatically during the era of vaccination, but trends in mortality are largely unknown. METHODS: US death certificates from 1990 to 2004 for which hepatitis A was listed as the underlying cause of death were analyzed. Average annual age-adjusted mortality rates during the prevaccine (1990-1995) and post vaccination recommendation (2000-2004) periods were compared using a Mantel Haenszel test of association. The number of deaths for which CLD was listed as a contributing cause was determined. RESULTS: Overall, 1436 deaths due to hepatitis A occurred, averaging 96 annually (range, 142 in 1995 to 54 in 2003). CLD contributed to nearly half of these deaths. Mortality rates paralleled incidence rates, beginning to decline in the mid-1990s and achieving low points in 2003 and 2004. Average rates were 32% lower in the post-vaccination recommendation period than in the prevaccine period (P < .01). The decline was more dramatic for states with (45%; P < .001) than without (23%; P = .002) recommendations. CONCLUSIONS: Hepatitis A mortality rates have declined over the past decade. CLD remains an important and preventable contributing cause of death due to hepatitis A. PMID- 18422441 TI - Double-blind study comparing 2 dosages of valacyclovir hydrochloride for the treatment of uncomplicated herpes zoster in immunocompromised patients 18 years of age and older. AB - A dosage of 1 g of valacyclovir 3 times per day (TID) for 7 days has already been shown to be superior to an oral dosage of 800 mg acyclovir 5 times per day for 7 days in immunocompetent individuals. The objective of this study was to assess the safety and efficacy of an oral dosage of valacyclovir, 1 g TID versus 2 g TID, for the treatment of herpes zoster in immunocompromised patients > or =18 years of age. The oral dosage schedule of 2 g of valacyclovir TID reaches acyclovir plasma levels similar to those achieved with intravenous acyclovir therapy given to immunocompromised patients (10 mg/kg every 8 h for 7 days). In this double-blind study, 87 immunocompromised patients with clinical evidence of localized herpes zoster were randomized to receive oral valacyclovir therapy for 7 days, either 1 g TID or 2 g TID, within 72 h after onset of zoster rash. Patients were seen and assessed for cutaneous healing, zoster-associated pain (ZAP), and/or zoster-associated abnormal sensations (ZAAS), up to 24 weeks. Participants in both arms of the study demonstrated similar median times to full crusting of the rash (8 days), and both dosages were safe and effective therapies for reduction of ZAP and ZAAS in the immunocompromised patient population. PMID- 18422442 TI - Comprehensive analysis of west nile virus-specific T cell responses in humans. AB - BACKGROUND: Cellular responses have been shown to play a role in immune control and clearance of West Nile virus (WNV) in murine models. However, little is known about the immunogenic regions of the virus or the phenotype of responding T cells in human infection. METHODS: Frozen peripheral blood mononuclear cells (PBMCs) from 35 WNV-infected blood donors were screened for virus-specific T cell responses by an interferon-gamma (IFN-gamma) enzyme-linked immunosorbent spot assay that used 452 overlapping peptides spanning all WNV proteins. More-detailed phenotypic studies were performed on subjects with high-magnitude T cell responses. RESULTS: In individuals with identified responses, the total number of recognized WNV peptides ranged from 1 to 9 (median, 2 peptides), and the overall magnitude of responses ranged from 50 to 4210 spot-forming cells (SFCs) per 10(6) PBMCs (median, 130 SFCs/10(6) PBMCs). A subset of 8 frequently recognized peptides from the regions of the genome encoding membrane, envelope, and nonstructural 3 and 4b proteins was identified. Phenotypic study of the highest magnitude WNV-specific T cell responses revealed that most were mediated by CD8+ cells that expressed perforin and/or granzyme B. CONCLUSIONS: These findings are the first to define the breadth and characteristics of the human T cell response to WNV and have implications for candidate vaccine design and evaluation. PMID- 18422443 TI - Virological and immunological correlates of mother-to-child transmission of cytomegalovirus in The Gambia. AB - BACKGROUND: Cytomegalovirus (CMV) is the most common congenital infection and can follow primary and recurrent maternal infection. We studied correlates of vertical transmission of CMV in The Gambia, where most children acquire CMV during the first year of life. METHODS: A cohort of 281 mothers and infants was recruited at birth. Infants were prospectively followed up for CMV infection during the first year of life. Excretion of CMV and antiviral immune response were studied at birth in mothers of children infected in utero, early during infancy, or late during infancy or not infected at 1 year of age. RESULTS: Congenital infection was diagnosed in 3.9% of newborns, and 85% of children were infected by 1 year. Excretion of CMV in colostrum or in the genital tract was more common in mothers of congenitally (100%) or early infected children (48%) than in mothers of late-infected (20%) or uninfected children (27%). Higher rates of viral excretion were associated with significantly higher levels of serum anti CMV immunoglobulin G and higher frequencies of CMV-specific CD4+ T cells. CONCLUSION: In the context of recurrent maternal infection, transmission of CMV in utero and during early postnatal life is associated with excretion of the virus in colostrum and the genital tract. PMID- 18422444 TI - Oseltamivir prophylactic regimens prevent H5N1 influenza morbidity and mortality in a ferret model. AB - BACKGROUND: Current oseltamivir prophylactic regimens may not be as effective against highly pathogenic H5N1 influenza viruses as they are against less pathogenic strains. An optimal regimen is urgently needed. METHODS: Ferrets were given the neuraminidase inhibitor oseltamivir orally for 10 days (5 or 10 mg/kg once daily or 2.5 or 5 mg/kg twice daily). Prophylaxis was initiated 1 day before infection, and oseltamivir was given 4 h before the ferrets were inoculated with a lethal dose of A/Vietnam/1203/04 (H5N1) influenza virus. RESULTS: At a dose of 5 mg/kg once daily, oseltamivir prevented death but not clinical signs of infection in ferrets; severe pathology was observed in the lungs, brain, and liver. At 10 mg/kg once daily, oseltamivir reduced clinical symptoms and systemic virus replication, but pathology was observed in the internal organs. The best results were obtained at a dose of 2.5 or 5 mg/kg given twice daily. Both regimens resulted in 100% survival and the absence of clinical symptoms, systemic virus spread, and organ pathology. Serum antibody titers were comparable across regimens and were sufficient to protect against rechallenge. CONCLUSIONS: An increased dose of oseltamivir or twice-daily administration effectively protects ferrets against morbidity and mortality caused by H5N1 infection and does not interfere with the development of protective antibodies against subsequent H5N1 infection. PMID- 18422447 TI - Expression of major histocompatibility complex class I chain-related molecule A, NKG2D, and transforming growth factor-beta in the liver of humans with alveolar echinococcosis: new actors in the tolerance to parasites? AB - BACKGROUND: Echinococcus multilocularis growth and persistent granuloma, which lead to the development of the severe parasitic disease alveolar echinococcosis (AE), might be caused by abnormal expression of stress-induced proteins, with subsequent abnormalities in T cell activation. Similar to its involvement in tumors, the NKG2D-major histocompatability complex class I chain-related molecules A and B (MICA/B) signaling system could be involved in host-parasite interactions; however, its involvement in helminthic diseases has never been studied. METHODS: We studied MICA/B, NKG2D, and transforming growth factor-beta (TGF-beta) expression on liver sections and measured levels of soluble MICA in serum samples obtained from patients with progressive AE. Livers from healthy and cirrhotic subjects were studied as controls. RESULTS: Expression of MICA/B proteins was strongly enhanced in the hepatocytes and endothelial and bile duct cells; in the CD68+ cells of the periparasitic infiltrate, especially epithelioid and giant cells; and, also, in the metacestode germinal layer. Strong expression of MICA/B in the liver contrasted with low numbers of NK cells and lack of expression of NKG2D on the numerous CD8+ T lymphocytes of the periparasitic infiltrate, as well as with the absence of soluble MICA in serum. TGF-beta was strongly expressed by most of the infiltrating lymphocytes. CONCLUSIONS: Sustained expression of MICA/B molecules and TGF-beta might lead to modulation of NKG2D with subsequent inhibition of NKG2D-dependent cytotoxicity. Abnormalities of this signaling system could contribute to parasitic evasion of the host's immunity. PMID- 18422445 TI - Scaling up antiretroviral therapy in South Africa: the impact of speed on survival. AB - BACKGROUND: Only 33% of eligible human immunodeficiency virus (HIV)-infected patients in South Africa receive antiretroviral therapy (ART). We sought to estimate the impact of alternative ART scale-up scenarios on patient outcomes from 2007-2012. METHODS: Using a simulation model of HIV infection with South African data, we projected HIV-associated mortality with and without effective ART for an adult cohort in need of therapy (2007) and for adults who became eligible for treatment (2008-2012). We compared 5 scale-up scenarios: (1) zero growth, with a total of 100,000 new treatment slots; (2) constant growth, with 600,000; (3) moderate growth, with 2.1 million; (4) rapid growth, with 2.4 million); and (5) full capacity, with 3.2 million. RESULTS: Our projections showed that by 2011, the rapid growth scenario fully met the South African need for ART; by 2012, the moderate scenario met 97% of the need, but the zero and constant growth scenarios met only 28% and 52% of the need, respectively. The latter scenarios resulted in 364,000 and 831,000 people alive and on ART in 2012. From 2007 to 2012, cumulative deaths in South Africa ranged from 2.5 million under the zero growth scenario to 1.2 million under the rapid growth scenario. CONCLUSIONS: Alternative ART scale-up scenarios in South Africa will lead to differences in the death rate that amount to more than 1.2 million deaths by 2012. More rapid scale-up remains critically important. PMID- 18422446 TI - Ascaris lumbricoides-induced interleukin-10 is not associated with atopy in schoolchildren in a rural area of the tropics. AB - BACKGROUND: In areas where intestinal helminth infections are endemic, infections by these parasites may protect against skin test-measured reactivity to allergens, and it has been suggested that interleukin (IL)-10 may mediate this effect. This study investigated whether IL-10 and populations of IL-10+ T cells may modulate atopy in children living in an area where intestinal helminth infections are endemic. METHODS: Ecuadorian schoolchildren from rural communities were assessed for skin test-measured reactivity to Periplaneta americana allergen and Dermatophagoides pteronyssinus allergen. Blood samples were collected from 39 skin test-positive and 41 skin test-negative children, and peripheral-blood leukocytes were cultured in the presence of Ascaris lumbricoides antigen, to measure IL-10 protein and the frequency of T cell populations expressing intracellular IL-10. We also investigated whether these immunological factors affected the association between allergen-specific IgE and skin test-measured reactivity to aeroallergens. RESULTS: There was no evidence of association between the level of A. lumbricoides-induced IL-10 protein or IL-10+ T cells and skin test-measured reactivity to allergens. The association between allergen specific IgE and skin test-measured reactivity was not affected by the level of IL-10 protein or the frequency of IL-10+ T cells. CONCLUSIONS: The results of this study do not support the notion that IL-10 plays a role in modulating atopy in children living in a tropical area where intestinal helminth infections are endemic. PMID- 18422448 TI - Polymorphic and conserved targets of antibodies against Plasmodium falciparum during pregnancy. PMID- 18422450 TI - Persistent Borrelia burgdorferi infection after treatment with antibiotics and anti-tumor necrosis factor-alpha. PMID- 18422452 TI - Achieving a healthy weight gain during pregnancy. AB - This review uses the 1990 U.S. Institute of Medicine (IOM) gestational weight gain recommendations to examine the question, what is a healthy pregnancy weight gain? The relationship of gestational weight gain to infant size at birth; pregnancy, labor, and delivery complications; neonatal, infant, and child outcomes; and maternal weight and health outcomes in U.S. and European populations are discussed. Pregnancy weight gains within the IOM recommendations are associated with better outcomes. The possible exception is very obese women, who may benefit from weight gains less than the 7 kg (15 pounds) recommended. Only about 33% to 40% of U.S. women gain within IOM recommendations. Excessive gestational weight gain is more prevalent than inadequate gain. Women's gestational weight gains tend to follow the recommendations of health care providers. Current interventions demonstrate efficacy in influencing gestational weight gain in low-income women with normal and overweight body mass index in the United States and obese women in Scandinavia. PMID- 18422453 TI - Laboratory-based surveillance of paratyphoid fever in the United States: travel and antimicrobial resistance. AB - BACKGROUND: The incidence of paratyphoid fever, including paratyphoid fever caused by antimicrobial-resistant strains, is increasing globally. However, the epidemiologic and laboratory characteristics of paratyphoid fever in the United States have never been studied. METHODS: We attempted to interview all patients who had been infected with laboratory-confirmed Salmonella serotypes Paratyphi A, Paratyphi B, or Paratyphi C in the United States with specimens collected from 1 April 2005 through 31 March 2006. At the Centers for Disease Control and Prevention (CDC), isolates underwent serotype confirmation, antimicrobial susceptibility testing, and pulsed-field gel electrophoresis typing. RESULTS: Of 149 patients infected with Salmonella Paratyphi A, we obtained epidemiologic information for 89 (60%); 55 (62%) of 86 were hospitalized. Eighty-five patients (96%) reported having travel internationally, and 80 (90%) had traveled to South Asia. Of the 146 isolates received at the CDC, 127 (87%) were nalidixic acid resistant; nalidixic acid resistance was associated with travel to South Asia (odds ratio, 17.0; 95% confidence interval, 3.8-75.9). All nalidixic acid resistant isolates showed decreased susceptibility to ciprofloxacin (minimum inhibitory concentration, > or = 0.12 microg/mL). Of 49 patients infected with Salmonella Paratyphi B, only 12 (24%) were confirmed to have Paratyphi B when tested at the CDC. Four (67%) of 6 patients were hospitalized, and 5 (83%) reported travel (4 to the Andean region of South America). One case of Salmonella Paratyphi C infection was reported in a traveler to West Africa with a urinary tract infection. CONCLUSIONS: Physicians should be aware of the increasing incidence of infection due to Salmonella Paratyphi A and treatment options given its widespread antimicrobial resistance. A paratyphoid fever vaccine is urgently needed. Continued surveillance for paratyphoid fever will help guide future prevention and treatment recommendations. PMID- 18422454 TI - HIV and the kidney: a spotlight on racial disparities. PMID- 18422456 TI - Leishmania infantum promastigotes reduce entry of HIV-1 into macrophages through a lipophosphoglycan-mediated disruption of lipid rafts. AB - Visceral leishmaniasis is now recognized as an opportunistic disease in patients infected with human immunodeficiency virus type 1 (HIV-1). We report here that Leishmania infantum promastigotes enhance HIV-1 replication in monocyte-derived macrophages (MDMs) at late time points in the virus growth curve but also that, surprisingly, a reduction in HIV-1 production is seen during the initial days after infection. This early effect is caused by a Leishmania-mediated inhibition of virus entry into MDMs through the action of lipophosphoglycan (LPG), the major promastigote surface glycolipid. The impact of LPG in the observed phenomenon was confirmed using LPG-defective lpg1-/- knockout mutant promastigotes. Our results suggest that the LPG-mediated effect results from the disruption of lipid rafts. Altogether, these findings suggest that the presence of Leishmania within the same cellular microenvironment leads to 2 opposite, time-dependent effects on HIV 1 replication. Leishmania and HIV-1 can thus establish complex interactions in their common natural host cells. PMID- 18422455 TI - The efficacy of clarithromycin and the bicyclolide EDP-420 against Mycobacterium avium in a mouse model of pulmonary infection. AB - Lung disease caused by Mycobacterium avium complex (MAC) is increasing in prevalence. MAC disease occurs in patients with chronic preexisting obstructive pulmonary diseases but is also diagnosed in individuals with no history of lung pathology or identifiable immune defect. Histologically, the disease is characterized by either the development of nodular granulomatous lesions in the peribronchial region or cavitary peripheral disease in smokers. Response to long term treatment is poor. Limited comparative-efficacy data on treatment exist. A model that resembles nodular MAC disease was established in C57 (bg+/bg+) mice infected intranasally. Therapy with clarithromycin, a compound commonly used to treat MAC disease, was evaluated in parallel with treatment using a new bicyclolide, EDP-420, that achieves high levels of intrapulmonary concentrations. Although clarithromycin administered daily resulted in a reduction in the bacterial load in the lung, EDP-420 administered either daily or twice a week was significantly more effective. These results suggest that this animal model can be used to evaluate novel regimens against MAC disease and that compounds with high concentration in the lung might have a significant impact on the outcome of MAC lung disease. PMID- 18422457 TI - The enterobacterial common antigen-like gene cluster of Haemophilus ducreyi contributes to virulence in humans. AB - Haemophilus ducreyi 35000HP contains a cluster of homologues of genes required for the synthesis of enterobacterial common antigen (ECA), suggesting that H. ducreyi may express a putative ECA-like glycoconjugate. WecA initiates the synthesis of ECA by transferring N-acetylglucosamine to undecaprenyl-P, to form lipid I. A wecA mutant (35000HPwecA) was constructed, and 5 volunteers were inoculated at 3 sites with fixed doses of 35000HP on one arm and at 3 sites with varying doses of 35000HPwecA on the other arm. 35000HPwecA caused pustules to form at 3 sites inoculated with a dose 2.5-fold higher than that of 35000HP. However, at sites inoculated with similar doses of 35000HP and 35000HPwecA, pustules developed at 46.7% (95% confidence interval [CI], 23.3%-70.0%) of 15 parent-strain sites and at 8.3% (95% CI, 0.01%-23.6%) of 12 mutant-strain sites (P = .013). Thus, the expression of wecA contributes to the ability of H. ducreyi to cause pustules in humans. PMID- 18422458 TI - Chronic kidney disease incidence, and progression to end-stage renal disease, in HIV-infected individuals: a tale of two races. AB - BACKGROUND: Little is known about the racial differences in the incidence and progression of HIV-related chronic kidney disease (CKD) that underlie African American-white disparities in HIV-related end-stage renal disease (ESRD). METHODS: In a cohort in Baltimore, Maryland, we measured CKD incidence, glomerular filtration rate (GFR) slope, and progression to ESRD in 3332 African American and 927 white HIV-infected subjects. RESULTS: A total of 284 subjects developed CKD, 100 (35%) of whom subsequently developed ESRD. African American subjects were at slightly increased risk for incident CKD, compared with white subjects (hazard ratio [HR], 1.9 [95% confidence interval {CI}, 1.2-2.8]). However, once CKD had commenced, the African American subjects developed ESRD markedly faster than did the white subjects (HR, 17.7 [95% CI, 2.5-127.0]), and, correspondingly, their GFR decline after diagnosis of CKD was 6-fold more rapid (P < .001). In the subset of African American subjects for whom kidney-biopsy data were available, progression to ESRD was significantly faster than that in white subjects with CKD, irrespective of the presence of HIV-associated nephropathy. CONCLUSIONS: The results of this study suggest that African American white disparities in HIV-related ESRD are explained predominantly by a more aggressive natural disease history in African Americans and less by racial differences in CKD incidence. PMID- 18422460 TI - Protease inhibitor resistance update: where are we now? AB - The introduction of protease inhibitors (PIs) and highly active antiretroviral therapy in the mid-1990s dramatically altered the treatment of HIV infection, enabling suppression of viral replication to undetectable levels and preventing disease progression. Most PIs present a strong barrier against viral resistance; the accumulation of multiple mutations is often required to produce resistance. However, there is variability of resistance within the PI class, as demonstrated by the fact that some PIs require fewer mutations to confer resistance compared with others. Resistance to individual PIs as well as the development of broad cross-resistance to multiple agents in this class remain major challenges in clinical practice. Resistance to PIs may involve primary or secondary mutations in the protease gene in addition to mutations outside of protease in the gag cleavage and noncleavage sites. Primary mutations may be sufficient to confer resistance to select PIs. Secondary mutations may be required to produce resistance with some PIs, whereas other mutations may be compensatory, restoring activity of the viral protease or increasing the replicative capacity of the virus. Specific resistance patterns associated with individual PIs have been identified. Strategies to prevent PI cross-resistance and to manage its occurrence involve rational sequencing of PIs, ritonavir boosting to maintain a strong barrier against viral resistance, the use of newer PIs with activity against resistant viruses or unique resistance profiles, avoidance of PI combinations with overlapping resistance patterns, and application of knowledge of mutations associated with hypersusceptibility to other agents in this class. PMID- 18422459 TI - An open-label pilot study to determine the efficacy of lopinavir/ritonavir and tenofovir DF in the treatment of HIV-infected patients experiencing first virologic failure on a non-nucleoside-based regimen. PMID- 18422462 TI - Spectrum of adverse events after generic HAART in southern Indian HIV-infected patients. AB - To determine the incidence of clinically significant adverse events after long term, fixed-dose, generic highly active antiretroviral therapy (HAART) use among HIV-infected individuals in South India, we examined the experiences of 3154 HIV infected individuals who received a minimum of 3 months of generic HAART between February 1996 and December 2006 at a tertiary HIV care referral center in South India. The most common regimens were 3TC + d4T + nevirapine (NVP) (54.8%), zidovudine (AZT) + 3TC + NVP (14.5%), 3TC + d4T + efavirenz (EFV) (20.1%), and AZT + 3TC + EFV (5.4%). The most common adverse events and median CD4 at time of event were rash (15.2%; CD4, 285 cells/microL) and peripheral neuropathy (9.0% and 348 cells/microL). Clinically significant anemia (hemoglobin <7 g/dL) was observed in 5.4% of patients (CD4, 165 cells/microL) and hepatitis (clinical jaundice with alanine aminotransferase > 5 times upper limits of normal) in 3.5% of patients (CD4, 260 cells/microL). Women were significantly more likely to experience lactic acidosis, while men were significantly more likely to experience immune reconstitution syndrome (p < 0.05). Among the patients with 1 year of follow-up, NVP therapy was significantly associated with developing rash and d4T therapy with developing peripheral neuropathy (p < 0.05). Anemia and hepatitis often occur within 12 weeks of initiating generic HAART. Frequent and early monitoring for these toxicities is warranted in developing countries where generic HAART is increasingly available. PMID- 18422463 TI - ADVANCE study begins enrollment. PMID- 18422461 TI - Self-reported sexual dysfunction is frequent among HIV-infected persons and is associated with suboptimal adherence to antiretrovirals. AB - Increased occurrence of sexual dysfunction (SD) among patients treated with highly active antiretroviral therapy (HAART) has been reported. To assess prevalence of self-reported SD and to identify factors related to this alteration with special focus to its relationship with adherence behavior, we conducted an intercohort analysis among HIV-infected persons treated with HAART. In an anonymous questionnaire investigating HAART nonadherence, patients were asked to report the occurrence of dysfunction in sexual activity over the previous 4 weeks. Among 612 participants, 125 (21%) reported some degree of SD. "Moderate"/"severe" alterations were reported in 6% and were independently associated with self-reported worsening of viro-immunological parameters (OR 3.90; 95% CI 1.08-14.18), higher symptom score (OR 1.13; 95% CI 1.05-1.22), and reporting abnormal fat accumulation (OR 4.33; 95% CI 1.55-12.11). Furthermore, nonadherent persons had an increased risk of SD (OR 3.44; 95% CI 1.30-9.08). In conclusion, patients' perceived SD represents a relevant problem for HIV-infected persons treated with antiretrovirals and is strongly associated with suboptimal HAART adherence. PMID- 18422464 TI - Five-year data released for entecavir. PMID- 18422465 TI - Different behavior of meniscal cells in collagen II/I,III and Hyaff-11 scaffolds in vitro. AB - The purpose of this study was to evaluate the behavior of ovine meniscal cells seeded on biomaterials made from collagen and hyaluronan, respectively. Ovine meniscal cells were isolated from the medial menisci of stifle joints, expanded in monolayer culture, and seeded on scaffolds made of collagen type II and I/III and a hyaluronan derivative (Hyaff-11). The samples were cultured for 12 h and 7, 14, 21, and 28 days. Histological analysis, electron microscopy, biochemical assays for glycosaminoglycans (GAGs) and DNA, and reverse transcriptase polymerase chain reaction analysis for collagens were performed. The cells attached well to both biomaterials and produced tissue-specific proteins, such as GAG and collagen type I, over a period of 28 days. Differences between the biomaterials were seen with respect to cell distribution, cell morphology, and the dynamics of GAG synthesis. The results show that ovine meniscal cells express their phenotype in both biomaterials. In terms of biology, collagen and hyaluronan are both suitable for tissue engineering in meniscal regeneration. It remains to be determined which scaffold possesses adequate biomechanical properties for successful in vivo application. PMID- 18422466 TI - The ENHANCE Study: an unusual publication of trial data raises questions beyond ezetimibe. AB - BACKGROUND: Over the past 2 years two unsuccessful clinical trials of lipid modifying agents, the ILLUMINATE Study for raising high-density lipoprotein and the ENHANCE Study for lowering low-density lipoprotein, have been reported. These results have surprised the field and led to a reassessment of the scientific and clinical basis for the overall mechanisms. OBJECTIVE: This paper aimed to consider the impact of the two trials, particularly the ENHANCE Study, on drug discovery and developments in dyslipidaemia and atherosclerosis. METHODS: The ILLUMINATE Study, which was a trial of the cholesteryl ester transfer protein inhibitor torcetrapib is briefly reviewed, followed by a more detailed discussion of the press release of the ENHANCE Study with ezetimibe, a cholesterol absorption inhibitor. The reaction of opinion leaders in the field with respect to low-density lipoprotein-lowering strategies and imaging methods is discussed. CONCLUSION: Even when accessible surrogate end-points such as plasma lipoprotein levels are available, the development of anti-atherosclerotic drugs acting by new mechanisms may be becoming more challenging. Imaging studies as an intermediate between early clinical studies and major outcome trials may be the way forward. PMID- 18422467 TI - Recommendations and treatment strategies for the management of acute ischemic stroke. AB - BACKGROUND: Stroke is one of the leading causes of mortality and disability worldwide. From the establishment of the penumbra concept, ischemic stroke has been recognized as a dynamic process and two main therapeutic strategies have been designed: one that tries to reopen the occluded artery and the second aims to protect the penumbra brain tissue until the physiologic mechanisms-or the treatment-stop the ischemia. OBJECTIVE: To review the most recent, high-quality evidence for acute stroke treatment. METHODS: Systematic review of relevant published studies focused in several aspects of acute ischemic stroke management, from neuroprotection to thrombolysis. CONCLUSIONS: After the publication of NINDS rt-PA study, the classical nihilistic approach to ischemic stroke started to change and thrombolytic treatment was approved in the treatment of patients with acute ischemic stroke presenting within 3 h from onset of symptoms. Advances in this field are proceeding on several fronts, including the use of next-generation plasminogen activators and glycoprotein IIb/IIIa inhibitors, refined patient selection with advanced magnetic resonance imaging sequences, endovascular approaches to thrombolysis and thrombectomy, and adjuvant use of ultrasound. Abrupt deprivation of oxygen and glucose to neuronal tissues elicits a series of pathologic cascades, leading to the spread of neuronal death. Of the numerous pathways identified, excessive activation of glutamate receptors, accumulation of intracellular Ca(2+) cations, abnormal recruitment of inflammatory cells, excessive production of free radicals and initiation of pathologic apoptosis are believed to play critical roles in ischemic damage, especially in the penumbral zone. Several neuroprotective agents designed to block these cascades have been investigated in animal models of cerebral ischemia and numerous agents have been found to reduce infarct size. However, translation of neuroprotective benefits from the laboratory bench to the emergency room has not been successful. Other measures, such as the relevance of body position in the acute phase of stroke, anticoagulant and antiplatelet agents or the effects of statins and antihypertensive therapy, are discussed in this paper, with an overview of the relevance of stroke units. PMID- 18422468 TI - Pleiotropic effects of thiazolidinediones. AB - BACKGROUND: Insulin resistance and hyperglycemia characterize type 2 diabetes mellitus. Type 2 diabetes mellitus is usually accompanied by concomitant disorders, such as dyslipidemia, hypertension and atherosclerosis. Thiazolidinediones are antidiabetic drugs that increase insulin sensitivity by activating the peroxisome proliferator-activated receptor gamma. There is evidence that thiazolidinediones exert a number of pleiotropic effects that may play an important role in the treatment of type 2 diabetes mellitus. OBJECTIVE: The purpose of the present article was to review the 'pleiotropic' effects of thiazolidinediones (i.e., their effects beyond glucose lowering). METHODS: The study involved searching PubMed up to February 2008 using relevant keywords. CONCLUSIONS: Thiazolidinediones favorably alter fat distribution and improve cardiovascular risk factors, such as blood pressure, inflammation markers and uric acid and they may also delay the progression of atherosclerosis. The effects on the lipid profile differ between the two thiazolidinediones studied with pioglitazone having more positive effects compared with rosiglitazone. Furthermore, thiazolidinediones improve diabetic complications, such as diabetic nephropathy and non-alcoholic fatty liver disease. Thiazolidinediones may also play a role in other diseases, such as polycystic ovary syndrome. These pleiotropic effects may prove to be clinically relevant. There has been recent debate about the possible differences between the two thiazolidinediones in terms of cardiovascular disease outcome. In this context, differences in the lipid effects between the two drugs may be relevant. PMID- 18422469 TI - Pharmacogenetics: potential role in the treatment of diabetes and obesity. AB - BACKGROUND: Common genetic variation is associated with increased risk of common metabolic diseases such as type 1 and type 2 diabetes, and obesity. Increasing experience with genetic association studies has led to an understanding of the large sample sizes required to detect a weak to moderate genetic predisposition to disease, the need to reproduce such associations in independent cohorts, and the statistical criteria required to detect a true association. This approach has been used successfully to identify disease-associated gene variation usually in representative populations of large numbers. OBJECTIVE: To review the current understanding of how common genetic variation influences predisposition to, and treatment of, metabolic disease. METHODOLOGY: Review of scientific literature. RESULTS: While there has been progress in understanding how genetic variation predisposes to diabetes and obesity, and how candidate genes may alter drug response, several caveats limit the interpretation and significance of pharmacogenetic studies published to date: those caveats typically include relatively small numbers of participants and choice of endpoints in determining gene-associated differences in response, which may not be clinically significant or relevant as a biomarker or predictor of a desired clinical effect. The genetic variants studied at a given locus are often limited in number and may not represent a comprehensive map of the region under study. CONCLUSIONS: The pharmacogenetic associations in diabetes and obesity that have been reported to date have had limited impact on the choice of individual treatments. We perceive, however, that this field is in its infancy in these multifactorial metabolic diseases, and with further advances and future drug intervention trials designed in a way that allows a more clear interpretation of the impact of genetic variation on differences in drug response in obesity and diabetes, it is anticipated that pharmacogenetics will have a significant impact on individualizing medical care. PMID- 18422470 TI - Cardiovascular risk in rheumatoid arthritis: effects of anti-TNF drugs. AB - BACKGROUND: Patients with rheumatoid arthritis (RA) have an increased risk of atherosclerotic cardiovascular disease which cannot be explained by traditional cardiovascular risk factors alone. Atherosclerosis is considered an inflammatory condition and inflammation experienced in RA may contribute to accelerated atherosclerosis. Thus, it should be hypothesized that treatment with antitumor necrosis factor alpha (anti-TNF-alpha), TNF-alpha being a pivotal component of the inflammatory cascade, may decrease concomitantly intra-articular inflammation and vessel inflammation. OBJECTIVE: The purpose of this review is to examine the data regarding cardiovascular mortality and morbidity in RA and the evidence available to date evaluating the influence of anti-TNF-alpha treatments in RA on the occurrence of cardiovascular events, on surrogate markers of atherosclerosis and classical cardiovascular risk factors. METHODS: Clinical trials, original studies and review articles were identified from a Medline search (1998 - December 2007). Articles in English were reviewed, with emphasis on those containing assessments of cardiovascular effects (i.e., biological, structural, clinical) of anti-TNF-alpha drug. CONCLUSION: The suppression of systemic inflammation favoring atherosclerosis may lead to an improvement in cardiovascular prognosis in inflammatory disorders. Thus, reduction of inflammatory joint disease in RA with anti-TNF-alpha therapy, as probably with any powerful disease-modifying antirheumatic drugs, seems to be, at least in part, associated with concomitant reduction of the risk of cardiovascular events. PMID- 18422471 TI - A review of irritable bowel syndrome and an update on therapeutic approaches. AB - BACKGROUND: Irritable bowel syndrome is a common disorder that is associated with a significant impact on both affected individuals and society. While the pathophysiology of irritable bowel syndrome remains unknown, knowledge regarding the normal and abnormal functions of the gut and its complex interactions with the body's nervous systems continue to shed light on the multifactiorial origins of irritable bowel syndrome symptoms. OBJECTIVE: This article provides an overview of the current knowledge on the therapeutic approaches to irritable bowel syndrome. METHODS: A search of the online bibliographic databases MEDLINE and EMBASE was performed in order to identify all relevant articles published between 1980 and 2008. The search was enhanced with the use of a medical librarian. Bibliographies from potentially relevant articles were manually searched. RESULTS/CONCLUSIONS: The therapeutic options for irritable bowel syndrome are rapidly evolving beyond traditional symptom-based therapies, such as fiber, antispasmodics, antidiarrheals and laxatives and are moving towards agents with organ-specific receptor selectivity directed, in many cases, at specific gastrointestinal functions. PMID- 18422472 TI - Food allergies and hypersensitivity: a review of pharmacotherapy and therapeutic strategies. AB - BACKGROUND: Food allergy is a major cause of life-threatening hypersensitivity reactions. Food-induced anaphylaxis is the most common reason for someone to present to the emergency department for an anaphylactic reaction. At present, the avoidance of the allergenic food is the only method of preventing further reactions for allergic patients. OBJECTIVE: With better characterization of allergens and an understanding of the immunologic mechanism involved in this reaction, investigators have developed several therapeutic modalities potentially applicable to the treatment and eventual prevention of food allergy. This review identifies and discusses the potential treatment options for food allergy that are under development. METHODS: Relevant articles are reviewed pertaining to the treatment of food allergy. RESULTS/CONCLUSION: Among the therapeutic options currently under investigation are anti-IgE therapy, peptide immunotherapy, traditional Chinese medicine, mutated protein immunotherapy, DNA immunization and immunization with immunostimulatory sequences linked to allergens. These novel forms of treatment for allergic disease hold promise for the safe and effective treatment of food-allergic individuals and the prevention of food allergy in the future. PMID- 18422473 TI - Developments in the pharmacotherapeutic management of spontaneous preterm labor. AB - BACKGROUND: Preterm birth is the major cause of perinatal mortality and morbidity in the developed world. OBJECTIVE: The aim of this study was to establish the importance of preterm birth and the huge healthcare costs involved and review the pathophysiology of preterm labor and the use of antepartum glucocorticoids, which are the main reason why tocolytics are used to prevent or delay preterm birth. The study also reviewed the range of tocolytics available, their mode of action and the evidence for their efficacy and fetomaternal safety. METHODS: An extensive review of the literature using well-recognized and accepted scientific search engines was employed. RESULTS/CONCLUSIONS: The perfect tocolytic does not exist. The evidence to support the use of magnesium sulfate as a tocolytic is poor. The use of beta-agonists is decreasing worldwide as clinicians move to nifedipine or atosiban, which are as effective but much safer. Although nifedipine is cheaper than atosiban and can be administered orally, the evidence to support atosiban is much superior to that of nifedipine and there have been recent safety concerns over nifedipine. PMID- 18422474 TI - The effect of anticoagulant pharmacotherapy on fracture healing. AB - BACKGROUND: There is in vitro and in vivo evidence that anticoagulants impair normal bone metabolism, and it is widely believed that this may impair fracture healing. However, there are only a few heterogeneous in vivo animal studies confirming this and the mechanisms are not fully understood. OBJECTIVE: To review the literature concerning the effects of anticoagulants on fracture healing, and to present current understanding of the mechanisms involved by reviewing in vivo studies of bone biology and in vitro studies of bone cells. METHODS: A systematic search of Medline and other databases was combined with manual searching of bibliographies of key papers to identify relevant studies in the English and German languages. CONCLUSION: There is strong evidence that warfarin, heparin and aspirin retard fracture healing. The preferential use of low molecular weight heparins is advocated to minimise this. Fondaparinux has not shown any impairment in vitro. Further studies of fondaparinux, the timing of anticoagulation therapy and the mechanisms of action of these agents are of paramount importance. PMID- 18422475 TI - Managing neuropathic pain with Sativex: a review of its pros and cons. AB - BACKGROUND: Although not new, the use of cannabis-based drugs for treating chronic pain patients is becoming a hot topic for pain physicians and other specialists due to the constant flow of medical information regarding this pharmacological therapy. Its indication is becoming more clearly targeted towards pain syndromes arising from nerve damage. The number of cases reported, clinical trials and reviews published on this subject exponentially increase year by year. A possible explanation for this may be the fact that neuropathic pain is a highly disabling symptom and, consequently, there is a demand from patients and health professionals for a definitive remedy to treat this pain. METHODS: Parallel to the number of articles on the effectiveness, recent articles describing the tolerability of cannabis-based drugs along with a more accurate characterisation of its side-effect profile and/or lack of effectiveness have been published, and they are placing a cautious stop for a more precise prescription of these medications. CONCLUSION: This article reviews the current knowledge on the use of Sativex for treating neuropathic pains of different origin, and analyses the balance between the advantages and drawbacks of this therapy. PMID- 18422476 TI - Experience with tenofovir disoproxil fumarate for antiretroviral therapy. AB - BACKGROUND AND RATIONALE/OBJECTIVE: Tenofovir disoproxil fumarate was approved in the US in 2001 and a few months later by the European Medicines Evaluation Agency for use in the EU, initially for treatment-experienced HIV patients with early virological failure. The following year the HIV indication was expanded to include antiretroviral treatment naive adults over 18 years of age. RESULTS AND CONCLUSIONS: Ongoing clinical trials of tenofovir disoproxil fumarate's safety and efficacy have confirmed its position as a long-term, safe, effective and convenient backbone component in combination antiretroviral therapy for HIV infected patients. With its favourable resistance profile, it is also an option for treatment-experienced patients. This article describes the efficacy and safety of tenofovir disoproxil fumarate from ongoing and recent clinical trials, and key aspects of its broad, clinical experience since its introduction. PMID- 18422477 TI - Imatinib mesilate for the treatment of gastrointestinal stromal tumour. AB - BACKGROUND: The molecular hallmark of gastrointestinal stromal tumours (GISTs), the mutation of the KIT gene, was discovered 10 years ago. GISTs have since been recognized as separate pathological entities among sarcomas, and have become a model for targeted treatment of solid tumours. Imatinib mesilate, which was approved in 2002 for the treatment of patients with advanced GIST, has dramatically changed the course of the disease. OBJECTIVE: This article will focus on the development of imatinib mesilate in the treatment of patients with GIST. METHODS: A Pubmed search was performed using the keywords 'imatinib', 'gastrointestinal stromal', 'GIST', 'KIT' and 'PDGFR'. Websites of the American Society of Clinical Oncology and the European Society of Medical Oncology were searched for data reported in abstract form at recent symposiums. Personal communications from opinion leaders were sought for additional information that might be relevant. RESULTS: Imatinib has changed the clinical course of patients with advanced GISTs and further development in the adjuvant setting as well as prospective assessment of predictive factors are the current focus of ongoing research. PMID- 18422478 TI - Treatment of advanced or recurrent colorectal cancer with irinotecan in Japan and elsewhere. AB - BACKGROUND: Dramatic prolongation of the survival of patients with advanced or recurrent colorectal cancer has been achieved by multi-drug chemotherapy using agents such as continuous 5-fluorouracil and leucovorin, irinotecan (CPT-11), L OHP, bevacizumab and cetuximab. OBJECTIVE: To explain the background of CPT-11 and to discuss its role in the treatment of advanced or recurrent colorectal cancer in Japan and elsewhere together with the authors' own experience. PMID- 18422479 TI - Experience of nitisinone for the pharmacological treatment of hereditary tyrosinaemia type 1. AB - BACKGROUND: Hereditary tyrosinaemia type 1 is a rare inherited metabolic condition, which leads to a fatal multisystemic disease in childhood. Since 1992, nitisinone - a compound developed from work on triketone herbicides - has become an effective pharmacological treatment by inhibiting the enzyme 4 hydroxyphenylpyruvate dioxygenase. OBJECTIVES: This review examines recent pharmacological and clinical literature on nitisinone, and assesses its impact as a pharmacological treatment for hereditary tyrosinaemia type 1. METHODS: English language literature from MedLine and EmBase for nitisinone was searched from 1990 to 2008 for all papers relevant to the use of nitisinone in hereditary tyrosinaemia type 1. CONCLUSIONS: Nitisinone can prevent the development of liver disease and significantly reduce the risk of developing hepatocellular carcinoma; however, vigorous surveillance for the development of HCC needs to be continued lifelong. PMID- 18422480 TI - Omega-3 fatty acid concentrates in the treatment of moderate hypertriglyceridemia. AB - BACKGROUND: Moderate hypertriglyceridemia is fairly common, and elevated triglycerides are a risk factor for coronary heart disease. The omega-3 fatty acids EPA and DHA have been shown to lower triglycerides in many clinical studies. Prescription omega-3 fatty acid concentrates (P-OM3) are indicated for use in people with very high triglycerides (> 500 mg/dl). Current guidelines recommend that triglycerides should be less < 150 mg/dl. OBJECTIVE: This review provides an overview of the use of omega-3 concentrates (both P-OM3 and over-the counter fish oil) to lower triglycerides in people who have moderate hypertriglyceridemia (triglycerides in the range of 150 - 500 mg/dl). The objectives were to examine clinical evidence, describe the magnitude of effects and predict future clinical use of P-OM3. METHODS: Published, peer-reviewed studies of omega-3 concentrates were included if they were placebo-controlled, double-blind, of sufficient size to demonstrate triglyceride lowering, and studied a population described as having a mean baseline triglyceride value of 150 - 500 mg/dl. Studies using the 4-g dose of P-OM3 were used to develop a model of percent triglyceride lowering as a function of baseline levels. RESULTS/CONCLUSIONS: P-OM3 are effective in reducing triglycerides by approximately 30% in this population and are likely to be combined with other drugs (e.g., statins) to treat combined dyslipidemia. PMID- 18422481 TI - Tolterodine for the treatment of overactive bladder. AB - BACKGROUND: The overactive bladder syndrome is a common condition affecting approximately 12% of men and women. It is extremely disturbing with a great impact on quality of life. Its treatment involves a combination of behavioural and pharmacological therapy. The latter includes antimuscarinic drugs such as tolterodine. OBJECTIVE: To review the safety and efficacy of tolterodine in the treatment of overactive bladder in comparison with other available antimuscarinic agents. METHODS: A Pubmed search was carried out differentiating randomised, clinical trials; longitudinal, retrospective studies; and metanalysis on the use of tolterodine for overactive bladder treatment. In the comparison with other antimuscarinic agents, only randomised, clinical trials were considered. RESULTS/CONCLUSION: Tolterodine is available as immediate- or extended-release formulations. It has been extensively evaluated with long-term, randomised trials for safety and efficacy showing a significant improvement in overactive bladder symptoms with an excellent tolerability profile. PMID- 18422482 TI - Oral pH-modified release budesonide for treatment of inflammatory bowel disease, collagenous and lymphocytic colitis. AB - BACKGROUND: Budesonide is a glucocorticoid with high topical glucocorticoid activity and low systemic availability. Oral pH-modified release budesonide has been investigated for the treatment of various inflammatory intestinal diseases. OBJECTIVE: To summarise the investigational data of oral pH-modified release budesonide for the treatment of Crohn's disease, ulcerative colitis, collagenous colitis and lymphocytic colitis. METHODS: Assessment of all published (full paper or meeting abstract) clinical studies with oral pH-modified release budesonide for the treatment of inflammatory intestinal diseases. RESULTS/CONCLUSION: Oral pH-modified release budesonide induces remission in mild-to-moderately active Crohn's disease of the ileum and/or ascending colon. This drug is useful to replace conventional systemic glucocorticoids in patients with ileocoecal Crohn's disease. Open-label studies showed efficacy in ulcerative colitis, but this finding has to be confirmed in controlled trials. Oral pH-modified release budesonide is also effective for the treatment of collagenous colitis and lymphocytic colitis. PMID- 18422483 TI - Functional characterization of ribosomal P1/P2 proteins in human cells. AB - The 'stalk' is a large ribosomal subunit domain that regulates translation. In the present study the role of the ribosomal stalk P proteins in modulating ribosomal activity has been investigated in human cells using RNA interference. A strong down-regulation of P2 mRNA and a drastic decrease in P2 protein in a stable human cell line was achieved using a doxycycline-inducible system. Interestingly, the amount of P1 protein was similarly decreased in these cells, in contrast with the expression of P1 mRNA. The loss of P1/P2 proteins produced a decrease in the growth rate of these cells, as well as an altered polysome pattern with reduced translation efficiency, but without affecting the free 40 S/60 S subunit ratio. A decrease in the ribosomal-subunit joining capacity was also observed. These data indicate that P1/P2 proteins modulate cytoplasmic translation by influencing the interaction between subunits, thereby regulating the rate of cell proliferation. PMID- 18422484 TI - Mouse neuroblastoma cells release prion infectivity associated with exosomal vesicles. AB - BACKGROUND INFORMATION: TSEs (transmissible spongiform encephalopathies) are neurodegenerative disorders affecting humans and animals. PrP(Sc), a conformationally altered isoform of the normal prion protein (PrP(C)), is thought to be the pathogenic agent. However, the biochemical composition of the prion agent is still matter of debate. The potential transmission risk of the prion agent through biological fluids has been shown, but the development of competitive diagnostic tests and treatment for TSEs requires a more comprehensive knowledge of the agent and the cellular mechanisms by which it is disseminated. With this aim, we initiated characterization of the prion agent and the pathways by which it can be propagated using the cellular model system neuroblastoma (N2a). RESULTS: The present study shows that N2a cells infected with scrapie release the prion agent into the cell culture medium in association with exosome like structures and viral particles of endogenous origin. We found that both prion proteins and scrapie infectivity are mainly associated with exosome-like structures that contain viral envelope glycoprotein and nucleic acids, such as RNAs. CONCLUSIONS: The dissemination of prions in N2a cell culture is mediated through the exosomal pathway. PMID- 18422485 TI - Effects of substitutions in the CXXC active-site motif of the extracytoplasmic thioredoxin ResA. AB - The thiol-disulfide oxidoreductase ResA from Bacillus subtilis fulfils a reductive role in cytochrome c maturation. The pK(a) values for the CEPC (one letter code) active-site cysteine residues of ResA are unusual for thioredoxin like proteins in that they are both high (>8) and within 0.5 unit of each other. To determine the contribution of the inter-cysteine dipeptide of ResA to its redox and acid-base properties, three variants (CPPC, CEHC and CPHC) were generated representing a stepwise conversion into the active-site sequence of the high-potential DsbA protein from Escherichia coli. The substitutions resulted in large decreases in the pK(a) values of both the active-site cysteine residues: in CPHC (DsbA-type) ResA, DeltapK(a) values of -2.5 were measured for both cysteine residues. Increases in midpoint reduction potentials were also observed, although these were comparatively small: CPHC (DsbA-type) ResA exhibited an increase of +40 mV compared with the wild-type protein. Unfolding studies revealed that, despite the observed differences in the properties of the reduced proteins, changes in stability were largely confined to the oxidized state. High-resolution structures of two of the variants (CEHC and CPHC ResA) in their reduced states were determined and are discussed in terms of the observed changes in properties. Finally, the in vivo functional properties of CEHC ResA are shown to be significantly affected compared with those of the wild-type protein. PMID- 18422486 TI - Palmitoylation of stathmin family proteins domain A controls Golgi versus mitochondrial subcellular targeting. AB - BACKGROUND INFORMATION: Precise localization of proteins to specialized subcellular domains is fundamental for proper neuronal development and function. The neural microtubule-regulatory phosphoproteins of the stathmin family are such proteins whose specific functions are controlled by subcellular localization. Whereas stathmin is cytosolic, SCG10, SCLIP and RB3/RB3'/RB3'' are localized to the Golgi and vesicle-like structures along neurites and at growth cones. We examined the molecular determinants involved in the regulation of this specific subcellular localization in hippocampal neurons in culture. RESULTS: We show that their conserved N-terminal domain A carrying two palmitoylation sites is dominant over the others for Golgi and vesicle-like localization. Using palmitoylation deficient GFP (green fluorescent protein) fusion mutants, we demonstrate that domains A of stathmin proteins have the particular ability to control protein targeting to either Golgi or mitochondria, depending on their palmitoylation. This regulation involves the co-operation of two subdomains within domain A, and seems also to be under the control of its SLD (stathmin-like domain) extension. CONCLUSIONS: Our results unravel that, in specific biological conditions, palmitoylation of stathmin proteins might be able to control their targeting to express their functional activities at appropriate subcellular sites. They, more generally, open new perspectives regarding the role of palmitoylation as a signalling mechanism orienting proteins to their functional subcellular compartments. PMID- 18422487 TI - Differential gene expression analysis of maize leaf at heading stage in response to water-deficit stress. AB - The whole-genomic gene-expression changes of maize (Zea mays L.) plants in response to water-deficit stress at the heading stage have not been previously studied. The present work utilized a maize oligonucleotide array ('57K', approximately 57000 sequences; http://www.maizearray.org/) representing more than 30000 unique genes, to profile transcriptome changes in maize leaves subjected to 1d (day) and 7d water-deficit stress. After 1d and 7d water-stress treatment, 195 and 1008 differential genes were identified respectively. One-third of 1d-water stress-induced genes had known or putative functions in various cellular signalling pathways, indicating that signal-transduction-related genes play important roles in the early responses of maize leaves to water stress. The 7d stress-regulated genes were involved in a broad range of cellular and biochemical activities. The most notable genes may function in compatible osmolyte metabolism, particularly in proline, sucrose, trehalose and raffinose metabolism in the leaves. The present study provided a valuable starting point for further elucidation of molecular mechanisms in the drought tolerance of maize plants. PMID- 18422488 TI - Elimination of competing hydrolysis and coupling side reactions of a cyclodextrin glucanotransferase by directed evolution. AB - Thermoanaerobacterium thermosulfurigenes cyclodextrin glucanotransferase primarily catalyses the formation of cyclic alpha-(1,4)-linked oligosaccharides (cyclodextrins) from starch. This enzyme also possesses unusually high hydrolytic activity as a side reaction, thought to be due to partial retention of ancestral enzyme function. This side reaction is undesirable, since it produces short saccharides that are responsible for the breakdown of the cyclodextrins formed, thus limiting the yield of cyclodextrins produced. To reduce the competing hydrolysis reaction, while maintaining the cyclization activity, we applied directed evolution, introducing random mutations throughout the cgt gene by error prone PCR. Mutations in two residues, Ser-77 and Trp-239, on the outer region of the active site, lowered the hydrolytic activity up to 15-fold with retention of cyclization activity. In contrast, mutations within the active site could not lower hydrolytic rates, indicating an evolutionary optimized role for cyclodextrin formation by residues within this region. The crystal structure of the most effective mutant, S77P, showed no alterations to the peptide backbone. However, subtle conformational changes to the side chains of active-site residues had occurred, which may explain the increased cyclization/hydrolysis ratio. This indicates that secondary effects of mutations located on the outer regions of the catalytic site are required to lower the rates of competing side reactions, while maintaining the primary catalytic function. Subsequent functional analysis of various glucanotransferases from the superfamily of glycoside hydrolases also suggests a gradual evolutionary progression of these enzymes from a common 'intermediate-like' ancestor towards specific transglycosylation activity. PMID- 18422489 TI - Translational science: past, present, and future. AB - The concept of translational science is at least 15 years old. However, in its most recent incarnation, it represents the identification of a funding category designed to encourage academic participation in a critical stage of the drug discovery and product development process. It is hoped that this will make the process both shorter and more efficient. In this review, the author first considers the historical development of the pharmaceutical R&D process. The place of translational science in the process, the scientific techniques involved, and aspects of the business environment necessary for its success are then considered. Translational science does not displace preclinical development. Both concepts are relevant to the paramount importance of successfully and expeditiously bridging the gap between preclinical science and clinical testing, "from bench to bedside." Translational science is particularly likely to stimulate biomarker research in the universities and related business community and will probably give a modest boost to early clinical testing and commercialization of discoveries within the academic setting. Whether there will be a consequent improvement in the quality and efficiency of the overall process remains to be seen. PMID- 18422490 TI - Increased efficiency for performing colony formation assays in 96-well plates: novel applications to combination therapies and high-throughput screening. AB - The colony formation assay (CFA) is the gold standard for measuring the effects of cytotoxic agents on cancer cells in vitro; however, in its traditional 6-well format, it is a time-consuming assay, particularly when evaluating combination therapies. In the interest of increased efficiency, the 6-well CFA was converted to a 96-well format using an automated colony counting algorithm. The 96-well CFA was validated using ionizing radiation therapy on the FaDu (human hypopharyngeal squamous cell) and A549 (human lung) cancer cell lines. Its ability to evaluate combination therapies was investigated by the generation of dose-response curves for the combination of cisplatin and radiation therapy on FaDu and A549 cells. The 96-well CFA was then transferred to a robotic platform for evaluating its potential as a high-throughput screening (HTS) readout. The LOPAC1280 library was screened against FaDu cells, and eight putative hits were identified. Using the 96-well CFA to validate the eight putative chemicals, six of the eight were confirmed, resulting in a positive hit rate of 75%. These data indicate that the 96-well CFA can be adopted as an efficient alternative assay to the 6-well CFA in evaluating single and combination therapies in vitro, providing a possible readout that could be used on a HTS platform. PMID- 18422491 TI - Profibrotic role of myostatin in Peyronie's disease. AB - INTRODUCTION: The primary histologic finding in many urologic disorders, including Peyronie's disease (PD), is fibrosis, mainly mediated by the transforming growth factor beta1 (TGFbeta1). AIM: To determine whether another member of the TGFbeta family, myostatin, (i) is expressed in the human PD plaque and normal tunica albuginea (TA), their cell cultures, and the TGFbeta1-induced PD lesion in the rat model; (ii) is responsible for myofibroblast generation, collagen deposition, and plaque formation; and (iii) mediates the profibrotic effects of TGFbeta1 in PD. METHODS: Human TA and PD tissue sections, and cell cultures from both tissues incubated with myostatin and TGFbeta1 were subjected to immunocytochemistry for myostatin and alpha-smooth muscle actin (ASMA). The cells were assayed by western blot, Real time-Polymerase chain reaction (RT-PCR), and ribonuclease protection. Myostatin cDNA and shRNA were injected, with or without TGFbeta1, in the rat penile TA, and plaque size was estimated by Masson. MAIN OUTCOME MEASURES: Myostatin expression in the human TA, the PD plaque, and their cell cultures, and myostatin effects on the PD-like plaque in the rat. RESULTS: A threefold overexpression of myostatin was found in the PD plaque as compared with the TA. In PD cells, myostatin expression was mainly in the myofibroblasts, and in the TA cells, it increased upon passage paralleling myofibroblast differentiation and was up-regulated by TGFbeta1. Myostatin or its cDNA construct increased the myofibroblast number and collagen in TA cells. Myostatin was detected in the TGFbeta1-induced PD-like plaque of the rat partly in the myofibroblasts, and in the TA. Myostatin cDNA injected in the TA induced a plaque and intensified the TGFbeta1 lesion, which was not reduced by myostatin shRNA. CONCLUSIONS: Myostatin is overexpressed in the PD plaque, partly because of myofibroblast generation. Although myostatin induces a plaque in the rat TA, it does not appear to mediate the one triggered by TGFbeta1, thus suggesting that both proteins act concurrently and that therapy should target their common downstream effectors. PMID- 18422492 TI - Does circumcision make a difference to the sexual experience of gay men? Findings from the Health in Men (HIM) cohort. AB - INTRODUCTION: The relevance of circumcision in preventing male-to-male sexual transmission of HIV is poorly understood, in particular because any potential beneficial effect could be diminished by the impact of circumcision on sexual behavior. AIM: We examined the impact of circumcision on sexual experience. METHODS: Univariate and multivariate logistic regressions were performed on data from 1,426 HIV-negative homosexually active men. MAIN OUTCOME MEASURES: We compared the sexual behaviors and preferences of circumcised with uncircumcised men, and men who were circumcised at infancy with those who were circumcised after infancy. RESULTS: Overall, 66% of men (N = 939) in the cohort were circumcised. After adjusting for age and ethnicity, we found no differences between circumcised and uncircumcised men in any insertive or receptive anal intercourse, difficulty using condoms, or sexual difficulties (e.g., loss of libido). Among the circumcised men, we compared those circumcised at infancy (N = 854) with those circumcised after infancy (N = 81). The majority cited phimosis (i.e., an inability to fully retract the foreskin) and parents' decision as the main reasons for circumcision after infancy. After adjusting for age and ethnicity, the men circumcised after infancy were more likely to practice any receptive anal sex (88% vs. 75%, P < 0.05) and to experience erection difficulties (52% vs. 47%, P < 0.05), but less likely to practice any insertive anal sex (79% vs. 87%, P < 0.05) and to experience premature ejaculation (15% vs. 23%, P < 0.05) than those circumcised at infancy. CONCLUSIONS: Our data suggest that overall circumcision status does not affect the HIV-negative gay men's anal sexual behaviors, experience of condom use, or likelihood of sexual difficulties. However, there is some suggestion of differences in sexual practices and preferences among circumcised gay men depending on the age at circumcision. In particular, gay men circumcised later are more likely to engage in and prefer receptive anal intercourse. PMID- 18422493 TI - Midodrine improves orgasm in spinal cord-injured men: the effects of autonomic stimulation. AB - INTRODUCTION: Orgasm is less frequent in men with spinal cord injury (SCI) than in able-bodied subjects, and is poorly understood. AIM: To assess the effect of autonomic stimulation on orgasm in SCI men using midodrine, an alpha1-adrenergic agonist agent. MATERIALS AND METHODS: Penile vibratory stimulation (PVS) was performed in 158 SCI men on midodrine as part of a treatment for anejaculation, after they failed a baseline PVS. A maximum of four trials were performed, weekly, with increasing doses of midodrine. MAIN OUTCOME MEASURE: The presence and type of ejaculation, orgasm experiences, and cardiovascular data were collected. RESULTS: Ejaculation either antegrade or retrograde was obtained in 102 SCI men (65%). Orgasm without ejaculation was reported by 14 patients (9%) on baseline PVS. Ninety-three patients (59%) experienced orgasm during PVS on midodrine. Orgasm was significantly related to the presence of ejaculation in 86 patients (84%), and more strikingly to antegrade ejaculation (pure or mixed with retrograde), i.e., in 98% of 70 patients. Orgasm was significantly more frequent in patients with upper motor neuron and incomplete lesions who present somatic responses during PVS. There was no effect of the presence of psychogenic erection. There was a significant increase in both systolic and diastolic blood pressure. Sixteen patients, mainly tetraplegics, developed intense autonomic dysreflexia (AD) that required an oral nicardipine chlorhydrate. CONCLUSIONS: Orgasm is the brain's cognitive interpretation of genital sensations and somatic responses, AD, and ejaculation. Intact sacral and T10-L2 cord segments are mandatory, allowing coordination between internal and external sphincters. Autonomic stimulation with midodrine enhances orgasm rate, mainly by creating antegrade ejaculation. PMID- 18422494 TI - Worry, desire, and sexual satisfaction and their association with severity of ED and age. AB - INTRODUCTION: This study evaluates worry about sexual and relationship functioning, sexual desire, and sexual satisfaction as indicators of sexual quality of life in men of different age groups suffering from mild to severe erectile dysfunction (ED). AIM: To increase insight in the mechanisms of some key indicators of sexual quality of life in different age groups. METHODS: The study sample consisted of 904 men with mild to severe ED. Mean age was 60.7 years (standard deviation [SD] = 12.4) and mean erectile function (EF) (International Index of Erectile Function [IIEF]) score was 14.5 (SD = 8.9). Multivariate analyses of variance were conducted to test the differences in outcomes among two age groups (65 65) and two levels of ED (16 16). MAIN OUTCOME MEASURES: We analyzed age, severity of ED, and interrelated indicators of sexual quality of life as worry about sexual or relationship functioning, sexual desire, and overall sexual satisfaction. Beyond, we aimed to see on which of these measures their interaction had specific independent effects. RESULTS: Younger men at a higher level of ED reported higher levels of worry about sexual and relationship functioning than older men with high ED. Older men reported slightly less sexual desire than younger men; also, more ED was associated with lower sexual desire. Finally, men with less severe ED reported higher sexual satisfaction than men with more severe ED, whereas at all levels of ED severity, older men reported more sexual satisfaction than younger men. CONCLUSIONS: Indicators of sexual quality of life among men with ED are poorer in those with more severe ED. However, younger men with severe ED worries concerning sexual functioning were more pronounced than among their older counterparts. Independent of the degree of severity of ED, older men indicated lower sexual desire but higher overall sexual satisfaction. Observed age-group differences were very small, however. PMID- 18422495 TI - The prevalence and risk factors of female sexual dysfunction in young korean women: an internet-based survey. AB - INTRODUCTION: In Korea, although male sexual problems have been relatively well addressed, few surveys have been conducted on female sexual dysfunction (FSD) in the general population. In the present study, we investigated the prevalence and identified the risk factors of sexual dysfunction in young Korean women. AIM: To evaluate the prevalence and to identify the risk factors of FSD in young Korean women. METHODS: A total of 47,000 women were initially approached. All received an e-mail requesting that they participate in a Web-based survey. The participants were asked to complete a questionnaire requesting detailed medical and sexual histories, which included the questions contained in the Korean version of the Female Sexual Function Index questionnaire. MAIN OUTCOME MEASURES: The prevalence of FSD in young Korean women in the different age groups and risk factors for developing FSD. RESULTS: A total of 504 women of average age 28.5 years (18-52 years) were evaluated during this survey. Setting the cutoff score for FSD using a receiver operating characteristic curve of our data as 25.0 points, 43.1% of women under 40 years old reported FSD. FSD was detected as a desire problem in 44.0% of women, an arousal problem in 49.0%, a lubrication problem in 37.0%, an orgasm problem in 32.0%, a satisfaction problem in 37.0%, and a pain problem in 34.6%. Risk factors for FSD as determined by logistic regression analysis were increasing age, a low frequency of sex, depression, a sexually abused history, and voiding dysfunction. CONCLUSIONS: The prevalence of FSD in Korean young women was common and comparable to those reported worldwide. PMID- 18422496 TI - Impact of premature ejaculation: the psychological, quality of life, and sexual relationship consequences. AB - INTRODUCTION: Premature ejaculation (PE) has been associated with a range of negative psychological effects, including anxiety, depression, and distress in men and their female partners. AIM: To review evidence of the psychosocial concomitants of premature ejaculation in recent observational studies, and to consider the psychosocial and quality of life outcomes associated with PE, including effects on the partner relationship. MAIN OUTCOME MEASURE: Psychosocial and quality of life consequences related to premature ejaculation. METHODS: A literature search was performed to retrieve publications relating to management or treatment of PE or male sexual dysfunction. Publications were included if they reported the impact of PE on the man, his partner or relationship, or the impact of male sexual dysfunction and included PE in the analysis. RESULTS: Eleven observational studies were selected. All these studies found evidence for an association between PE and adverse psychosocial and quality of life consequences, including detrimental effects on the partner relationship. Comparative analyses were restricted by major differences across the studies. CONCLUSIONS: PE significantly negatively impacts men and their partners and may prevent single men forming new partner relationships. Men are reluctant to seek treatment from their physicians, although they may be more encouraged to do so through their partner's support and the availability of effective treatments. There is a need for validated diagnostic screening criteria and validated, reliable, brief patient-reported outcome measures that can be used to assess men with PE and their partners. These factors would allow further studies with more complete and accurate assessment of the impact of PE. PMID- 18422497 TI - Salivary 8-OHdG: a useful biomarker for predicting severe ED and hypogonadism. AB - INTRODUCTION: Erectile and endothelial dysfunction are common pathologies of multiple cardiovascular risk factors and are considered longitudinal predictors of cardiovascular events. Oxidative stress and decreases in testosterone levels play an important role in the pathogenesis of endothelial dysfunction. AIM: We sought to determine whether the severity of erectile dysfunction (ED) was associated with individual levels of testosterone and oxidative stress, and whether treatment with a phosphodiesterase type 5 inhibitor could reduce oxidative stress and increase testosterone availability. METHODS: We evaluated the association of salivary 8-hydroxy-2'-deoxyguanosine (8-OHdG), salivary testosterone, International Index of Erectile Function-erectile function domain (IIEF-EF) scores, and Medical Outcome Study (MOS) 36-item Short-Form Healthy Survey (SF-36) questionnaires in 128 middle-aged male volunteers. We investigated the changes in testosterone levels, salivary 8-OHdG levels, IIEF-EF scores, and SF-36 scores in 20 ED patients (according to the IIEF-EF) who took 50 mg of sildenafil once a week for 6 months. MAIN OUTCOME MEASURES: IIEF-EF scores were used to assess ED severity. Antioxidant status was defined by salivary 8-OHdG. Salivary testosterone was used to evaluate serum bioavailable testosterone availability. RESULTS: Salivary 8-OHdG (OR = 9.88, 95% CI: 1.52-64.10), salivary testosterone (Odds ratio [OR] = 0.96, 95% CI: 0.93-0.98), and vitality on the SF 36, version 2 (SF-36 v2) (OR = 0.92, 95%CI: 0.84-0.98) were significantly associated with the severity of ED in healthy volunteers. Treatment with sildenafil for 6 months significantly increased the total serum testosterone (426.4 +/- 174.8 vs. 569.6 +/- 146.1 ng/dL, P = 0.021) and salivary testosterone levels (56.1 +/- 22.3 vs. 110.0 +/- 48.4 pg/mL, P < 0.001), whereas it decreased salivary 8-OHdG levels (2.30 +/- 0.23 vs. 0.90 +/- 0.05 ng/mL, P = 0.0046). CONCLUSIONS: Salivary 8-OHdG is a useful biomarker for predicting severe ED and hypogonadism in middle-aged men. Once-a-week treatment with sildenafil can have beneficial effects on men's health by decreasing oxidative stress and increasing testosterone levels. PMID- 18422498 TI - Opportunities and challenges of sexual health services among young people: a study in Nepal. AB - INTRODUCTION: It has been well documented that young people are more likely to engage in high-risk sexual activity. Appropriate understanding of safe sex, sexual practices, and related behaviors must recognize the importance of socioeconomic and cultural factors in prevention efforts related to HIV and other sexual transmitted infections (STIs). AIM: To examine and summarize the opportunities and challenges of sexual health services among young people in Nepal. MAIN OUTCOME MEASURES: Review of literature--assessing knowledge, attitudes, and understanding of sex, sexual health, and related sexual risk behaviors, among young people (15-24), in line with the current sociocultural and health service practices. METHODS: Medline, Cumulative Index to Nursing and Allied Health Literature (CINAHL), Science, Cochrane database, and Google were searched. Similarly, documents published at the WHO, United Nations Population Fund, United Nations Development Program, and at national/local level--Ministry of Health, National Center for AIDS, and STD Control were also assessed to access the relevant reports and articles. Published and gray articles were also reviewed. RESULTS: This study contends growing expansion of communication and transportation networks, urbanization, and urban in-migration is creating a different sociocultural environment, which is conducive to more social interactions between young girls and boys in Nepal. Rising age at marriage opens a window of opportunity for premarital and unsafe sexual activity among young people and this creates risks of unwanted pregnancy, STIs/HIV and AIDS. Socioeconomic, demographic, and cultural factors have been identified as encouraging factors for risk-taking behaviors among young people. CONCLUSIONS: Understanding safer sex and responsible sexual/reproductive behavior is important. Effective and appropriate interventions on sexual and reproductive health education directed at young people and the whole family, including fathers, could have significant effect on reducing risk and related risk practices in the context of Nepal. PMID- 18422499 TI - Diagnosis of hyperthyroidism in cats with mild chronic kidney disease. AB - OBJECTIVES: In cats with concurrent hyperthyroidism and non-thyroidal illnesses such as chronic kidney disease, total thyroxine concentrations are often within the laboratory reference range (19 to 55 nmol/l). The objective of the study was to determine total thyroxine, free thyroxine and/or thyroid-stimulating hormone concentrations in cats with mild chronic kidney disease. METHODS: Total thyroxine, free thyroxine and thyroid-stimulating hormone were measured in three groups. The hyperthyroidism-chronic kidney disease group (n=16) had chronic kidney disease and clinical signs compatible with hyperthyroidism but a plasma total thyroxine concentration within the reference range. These cats were subsequently confirmed to be hyperthyroid at a later date. The chronic kidney disease-only group (n=20) had chronic kidney disease but no signs of hyperthyroidism. The normal group (n=20) comprised clinically healthy senior (>8 years) cats. RESULTS: In 4 of 20 euthyroid chronic kidney disease cats, free thyroxine concentrations were borderline or high (> or =40 pmol/l). In the hyperthyroidism-chronic kidney disease group, free thyroxine was high in 15 of 16 cats, while thyroid-stimulating hormone was low in 16 of 16 cats. Most hyperthyroidism-chronic kidney disease cats (14 of 16) had total thyroxine greater than 30 nmol/l, whereas all the chronic kidney disease-only cats had total thyroxine less than 30 nmol/l. CLINICAL SIGNIFICANCE: The combined measurement of free thyroxine with total thyroxine or thyroid-stimulating hormone may be of merit in the diagnosis of hyperthyroidism in cats with chronic kidney disease. PMID- 18422500 TI - A retrospective study of the clinical presentation of 140 dogs and 39 cats with bacteraemia. AB - OBJECTIVES: To evaluate retrospective data from 140 dogs and 39 cats with positive blood cultures that were presented to the Clinic for Small Animal Medicine in Munich from 1995 to 2004. METHODS: The identity of bacteria isolated from blood cultures of dogs and cats with bacteraemia was determined, and clinical and laboratory findings and outcome of animals with Gram-negative versus Gram-positive bacteraemia were compared. RESULTS: Sepsis was diagnosed in 81.7 per cent of dogs and 59.5 per cent of cats with bacteraemia. Escherichia coli was isolated in one third of the animals. Dogs with bacteraemia more often showed monocytosis and increased alkaline phosphatase activity, while in cats, hyperglycaemia was found more commonly. Dogs with Gram-negative bacteraemia had hypoalbuminaemia significantly more often than dogs with Gram-positive bacteraemia, while among the remaining parameters, there were no statistically significant differences. CLINICAL SIGNIFICANCE: Not all dogs and cats with a positive blood culture met the criteria for sepsis. Bacteraemia caused by Gram positive versus Gram-negative bacteria cannot be distinguished based on clinical or laboratory parameters, and bacterial culture and susceptibility testing have to be performed for the right choice of antibiotic treatment. PMID- 18422501 TI - Efficacy and safety of cefovecin for the treatment of urinary tract infections in cats. AB - OBJECTIVES: To determine the efficacy and safety of cefovecin (Convenia); Pfizer Animal Health) in the treatment of urinary tract infections in cats. METHOD: A multi-centre, masked, randomised study was conducted in cats presenting with clinical signs indicative of urinary tract infections. Cephalexin (Rilexine); Virbac) administered orally twice daily at 15 mg/kg bodyweight for 14 days was compared with a single subcutaneous injection of cefovecin in cats. The primary efficacy parameter assessed was bacterial elimination of the pretreatment uropathogen. RESULTS: Four hundred and thirty-four cats were screened for urinary tract infections. One hundred and eighty-five cats were treated with either cefovecin (n=124) or cephalexin (n=61). Ninety-seven cats (22.2 per cent) had confirmed bacteriuria and 82 cats were included in efficacy analysis. Escherichia coli was eliminated in 76.7 per cent (23 of 30) of cefovecin-treated cats compared with 62.5 per cent (10 of 16) of cephalexin-treated cats. Cefovecin demonstrated statistical non-inferiority compared with cephalexin for bacterial elimination. There were no suspected adverse drug reactions attributable to treatment with cefovecin or cephalexin. CLINICAL SIGNIFICANCE: Cefovecin was demonstrated to be an effective and safe treatment for urinary tract infections. PMID- 18422502 TI - Colour M-mode tissue Doppler imaging in healthy cats and cats with hypertrophic cardiomyopathy. AB - OBJECTIVES: To determine whether decreased diastolic and systolic myocardial velocity gradient between the endocardium and the epicardium exist in the left ventricle of cats with hypertrophic cardiomyopathy. METHODS: Myocardial velocity gradient and mean myocardial velocities were measured by colour M-mode tissue Doppler imaging in the left ventricular free wall of 20 normal cats and 17 cats with hypertrophic cardiomyopathy. RESULTS: The peak myocardial velocity gradient (sec(-1)) during the first (E1) (5.71+/-1.75 versus 11.38+/-3.1, P<0.001) and second phase (E2) (3.09+/-1.53 versus 7.02+/-3.1, P=0.005) of early diastole and also the maximum early diastolic myocardial velocity gradient (Emax) (6.12+/-2.1 versus 10.76+/-3.2, P<0.001) were reduced in cats with hypertrophic cardiomyopathy compared with normal cats. Peak myocardial velocity gradient during early systole (Se) was lower in affected cats than in normal cats (6.26+/ 2.08 versus 8.67+/-2.83, P=0.006). Affected cats had a lower peak mean myocardial velocities (mm/s) during the two isovolumic periods (IVRb and IVCb) compared with normal cats (2.97+/-6.76 versus 12.74+/-5.5 and 22.28+/-9.96 versus 38.65+/-10.1, P<0.001, respectively). CLINICAL SIGNIFICANCE: This study shows that hypertrophic cardiomyopathy cats have decreased myocardial velocity gradient during both diastole and systole and also altered myocardial motion during the two isovolumic periods. Myocardial velocity gradients recorded by colour M-mode tissue Doppler imaging can discriminate between the healthy and diseased myocardium. PMID- 18422503 TI - Detection of Leishmania infantum by real-time PCR in a canine blood bank. AB - OBJECTIVES: Risk for transmission of Leishmania infantum from blood products has been largely demonstrated in human and veterinary literature. Appropriate screening of canine blood donors is important especially in an endemic area such as Barcelona (Spain). The purpose of this study was to evaluate the presence of L infantum DNA parasites by real-time quantitative PCR in our canine blood bank. METHODS: Samples from blood products obtained from 92 canine blood donors were assayed for L infantum by means of real-time PCR amplification and quantification. RESULTS: The prevalence of quantitative PCR-positive blood samples among healthy seronegative blood donors was 19.6 per cent. CLINICAL SIGNIFICANCE: The results of this study show that L infantum infection is common in canine blood donors and their blood products in an endemic area, despite a negative commercial serological screening for infectious diseases. Therefore, screening by PCR should be included in an integrated approach to evaluate L infantum infection among potential blood donors. PMID- 18422504 TI - Canine serum C-reactive protein detected by means of a near-patient test for human C-reactive protein. AB - OBJECTIVES: The aim of the study was to evaluate the reliability of a rapid human C-reactive protein near-patient slide reversed passive latex agglutination test (Randox) for the semi-quantitative determination of canine serum C-reactive protein. METHODS: The concentration of C-reactive protein was determined in 244 canine serum samples by an established automated immunoturbidimetric method and in various predilutions by a commercially available reversed passive latex agglutination test for human C-reactive protein. The results were compared to assess if the reversed passive latex agglutination test reflected the results of the established method with special emphasis on the reversed passive latex agglutination test's ability to identify samples characterised as positive or negative by the established method. RESULTS: The reversed passive latex agglutination test reflected the C-reactive protein concentration in canine serum samples at all the tested predilutions (undiluted, 1:4, 1:8 and 1:16). When applying a predilution of 1:8, the positive and negative analytical predictive values for discriminating between positive and negative samples (according to the established quantitative method) were high (0.94 [0.82 to 0.99] and 0.97 [0.93 to 0.99], respectively). CLINICAL SIGNIFICANCE: In conclusion, this near-patient test was able to reflect the serum C-reactive protein concentration in canine samples in a reliable and clinically useful manner and could be applicable for general practice for evaluating C-reactive protein levels in canine serum. PMID- 18422505 TI - Anaplastic and aggressive subcutaneous sarcoma in a seven-month-old dog. AB - A seven-month-old female, Italian hound which was presented with anorexia, dyspnoea and weight loss showed two subcutaneous masses at physical examination. Radiographs showed multiple opacities in the lungs, thorax, liver and spleen, and cytology showed discrete pleomorphic cells, with variable nuclear : cytoplasmic ratio, foamy cytoplasm and round to oval nuclei. Incisional biopsy histology showed the infiltrative proliferation of highly pleomorphic spindle to roundish large cells, discrete or arranged in a storiform pattern, with scattered multinucleated giant cells. Immunohistochemistry exhibited strong positivity for vimentin and mild labelling for alpha-smooth muscle actin and lysozyme; other mesenchymal or histiocytic lineage markers stained negatively. Because of the poor prognosis, the owner elected euthanasia. Post-mortem examination confirmed massive metastatic spread. Cytology, histology and immunohistochemistry suggested the diagnosis of anaplastic sarcoma with giant cells, although disseminated histiocytic sarcoma was a reliable differential diagnosis. The authors underline the difficulties in definitively categorising poorly differentiated sarcomas, even if a large panel of markers is applied. PMID- 18422506 TI - Vertebral heart scale in the beagle dog. AB - OBJECTIVE: To determine if Buchanan's vertebral heart scale can be applied to beagle dogs to establish reference values for this breed and to determine the possible influence of optimal lung inflation on the cardiac measurements. METHODS: The cardiac health of 19 beagle dogs was examined by echocardiography, electrocardiogram, non-invasive blood pressure measurement, complete blood count and serum chemistry profile. Right and left lateral recumbent radiographs of the thorax were obtained while the dogs were conscious and while under general anaesthesia with optimal lung inflation. The vertebral heart scale was measured in all radiographs. RESULTS: The mean (sd) vertebral heart scale in the beagle dogs was 10.3 (0.4) v, showing a significant difference from the mean vertebral heart scale of 9.7 (0.5) v in Buchanan's original study with dogs of different breeds. Degree of inspiration had no significant effect on the vertebral heart scale. CLINICAL SIGNIFICANCE: The vertebral heart scale provides the veterinarian with a possible way of objectively evaluating canine heart size without having to rely on echocardiography. However, the results should be interpreted critically and breed-specific differences should be taken into account, especially with values slightly above the reference range. PMID- 18422507 TI - Congenital hyposomatotropism in a domestic shorthair cat presenting with congenital corneal oedema. AB - A six-month-old, female, domestic shorthair cat was presented with a history of failure to grow and bilateral corneal opacity caused by corneal oedema. Congenital hyposomatotropism and possible secondary hypothyroidism were diagnosed on the basis of fasting serum levels of insulin-like growth factor-1 and thyroxine levels, respectively. These endocrinopathies are rare in the cat and have not been reported to cause ocular signs. The cat died during investigation of these diseases, and histopathological examination of the eyes showed significantly reduced corneal endothelial cell density and number of corneal epithelial cell layers when compared with age-matched healthy control corneas. These changes were implicated in the development of the corneal oedema. PMID- 18422508 TI - Evaluation of pharyngeal shape and size using anatomical optical coherence tomography in individuals with and without obstructive sleep apnoea. AB - This study compared shape, size and length of the pharyngeal airway in individuals with and without obstructive sleep apnoea (OSA) using a novel endoscopic imaging technique, anatomical optical coherence tomography (aOCT). The study population comprised a preliminary study group of 20 OSA patients and a subsequent controlled study group of 10 OSA patients and 10 body mass index (BMI) , gender- and age-matched control subjects without OSA. All subjects were scanned using aOCT while awake, supine and breathing quietly. Measurements of airway cross-sectional area (CSA) and anteroposterior (A-P) and lateral diameters were obtained from the hypo-, oro- and velopharyngeal regions. A-P : lateral diameter ratios were calculated to provide an index of regional airway shape. In all subjects, pharyngeal CSA was lowest in the velopharynx. Patients with OSA had a smaller velopharyngeal CSA than controls (maximum CSA 91 +/- 40 versus 153 +/- 84 mm(2); P < 0.05) but comparable oro- (318 +/- 80 versus 279 +/- 129 mm(2); P = 0.48) and hypopharyngeal CSA (250 +/- 105 versus 303 +/- 112 mm(2); P = 0.36). In each pharyngeal region, the long axis of the airway was oriented in the lateral diameter. Airway shape was not different between the groups. Pharyngeal airway length was similar in both groups, although the OSA group had longer uvulae than the control group (16.8 +/- 6.2 versus 11.2 +/- 5.2 mm; P < 0.05). This study has shown that individuals with OSA have a smaller velopharyngeal CSA than BMI-, gender- and age-matched control volunteers, but comparable shape: a laterally oriented ellipse. These findings suggest that it is an abnormality in size rather than shape that is the more important anatomical predictor of OSA. PMID- 18422509 TI - Personality traits and temporomandibular disorders in a group of children with bruxing behaviour. AB - The aim of the present study was to compare the personality pattern, the anxiety level and the temporomandibular disorders (TMD) between bruxist and non-bruxist children with mixed dentition. Fifty-two subjects, with a mean age of 9.45 years (range 8-11) were evaluated and classified as bruxist (n = 26), according to the American Academy of Sleep Medicine (AASM) and the presence of dental wear clinically visible, under the same conditions of artificial light and position. The control children (n = 26) did not present dental wear and did not accomplished all the AASM criteria. The personality pattern and the anxiety of the bruxist children were studied by means of the Children's Personality Questionnaire (CPQ) and the Conners' Parents Rating Scales (CPRS), respectively, and compared with the personality traits and the anxiety level of a non-bruxist population. The TMD were also evaluated using the Research Diagnostic Criteria for temporomandibular disorders (RDC/TMD) criteria. The data were analysed with the student's t-test, Fisher's exact test and chi-squared tests. A multivariated analysis was performed using a logistic regression with the stepwise likelihood ratio method. Compared with the controls, the bruxist children had significantly higher tension personality and were more anxiety prone. The bruxist children presented more TMD-related signs and symptoms than children in the control group. A strong correlation was found among bruxism, TMD, the high anxiety level and the high tension personality trait. PMID- 18422510 TI - Relationships between implant stability, image-based measures and nitric oxide levels. AB - For monitoring of dental implants, practitioners seem to use simultaneously a variety of image-based and laboratory measurements. This longitudinal study was conducted to analyse the possible relationships between nitric oxide (NO) content of peri-implant sulcus fluid (PISF), marginal bone levels and implant stability in terms of resonance frequency analysis (RFA) at immediately (IL) and conventionally (CL) loaded mandibular molar dental implants from the same individuals. Dental implants were placed in 10 patients who had first molar loss bilaterally in the mandibular area. One site of the patient was determined as IL and the other site was CL. PISF samples, marginal bone level and RFA were taken for all dental implants during a 12-month follow-up period. For both types of loading, there were no significant changes at implant stability (ISQ) and PISF volume during the whole study period. The lowest total nitrite levels were observed at the end of 12 months. Marginal bone gain of 0.22 and 0.09 mm was noticed between 6 and 12 months in IL and CL groups, respectively. During the study period, negative correlation was noted between radiographic bone level and ISQ at both IL and CL sites; however, this correlation reached a significant level only at 6 months in CL implant group (P = 0.015). Although not significant, marginal bone loss seems to have a negative relationship with the implant stability values regardless of the mode of loading. NO metabolism around IL and CL dental implants may not demonstrate a significantly different pattern. PMID- 18422511 TI - Relationship between dental implant stability determined by resonance frequency analysis measurements and peri-implant vertical defects: an in vitro study. AB - Resonance frequency (RF) analysis is a non-invasive and objective technique developed for implantology, where it measures the stability of the implant in the bone socket. A limited number of studies were performed to compare different stability measurement devices. The aim of the study was to evaluate the association between the implant stability and peri-implant vertical defect by using the new wireless RF analyser. Micro-textured rough-surfaced implants with diameters of 3.7 and 4.8 mm and with a length of 12 mm were used. The peak insertion torque was recorded with the help of the torque control system only during the implant placement. The peri-implant defects were created in millimetre increments ranging between 0 and 5 mm to the same extent on all implants. The RF analysis values were measured by using the RF analyser. Significant correlation was found between insertion torque (44.3 +/- 3 N cm) and implant stability quotient (ISQ) (all implants: r = 0.76, 3.7 mm diameter: r = 0.65, 4.8 mm diameter: r = 0.80). For 3.7 x 12-mm(2) implants, the mean ISQ values were 72.6 +/- 2.4, 69.8 +/- 2, 67.2 +/- 1.8, 64.3 +/- 1.9, 61.2 +/- 2 and 57.2 +/- 2.5 when peri-implant vertical defects were 0, 1, 2, 3, 4 and 5 mm; and the corresponding values were 76.5 +/- 2.2, 74.5 +/- 2.2, 72.3 +/- 2, 70 +/- 1.9, 67 +/- 1.9 and 63.4 +/- 2.3, respectively, for 4.8 x 12-mm(2) implants. All values were significantly different when compared with each other. The wireless RF analyser seems to be a suitable and reliable device to determine the implant stability. Peri-implant bone loss simulated by using acrylic models may result with a decrease in ISQ values for osseo-integrated implants measured by the RF analyser. PMID- 18422512 TI - Using computer-guided implantology in flapless implant surgery of a maxilla: a clinical report. AB - The current notion in implantology is to utilize techniques that can provide function, aesthetics and comfort with a minimally invasive surgical approach. The flapless implant surgery has been suggested to fulfil these requirements. Traditionally, the flapless implant surgery was performed by using a tissue punch technique, which may be potentially harmful because of the inherent blindness of the technique. Today, computer software programs and 3D radiographic techniques, such as CT technology, have been developed to provide the practitioner with precise implant placement planning. Although this sophisticated approach has many advantages, transferring this information to the surgical phase has only recently been developed. The purpose of this paper was to introduce a more predictable flapless approach for treatment of a patient's maxilla through principles of computer-assisted implantology. PMID- 18422513 TI - Evaluation of the use of and attitudes towards a face-bow in complete denture fabrication: a pilot questionnaire investigation in Chinese prosthodontists. AB - The aim of this study was to assess the use of and attitudes towards a face-bow among Chinese prosthodontists in complete denture fabrication. Thirty-six prosthodontists in two dental hospitals affiliated separately to two of the top four dental schools in China were included. A questionnaire was sent to them with 18 questions covering the following four aspects: (i) the education and experience background of the prosthodontists, (ii) their impression of the quality of the dentures, (iii) the use of and attitudes towards a face-bow in complete denture fabrication and (iv) their understanding of jaw relationship and occlusal adjustment in complete denture fabrication. Most (97%) experienced Chinese prosthodontists seldom used a face-bow in complete denture fabrication. Although they (92%) believed that more than 90% of their edentulous patients were satisfied with the complete dentures, 39% of them thought that not using a face bow might lower the quality of complete dentures and 94% believed that there was a difference between the quality of dentures fabricated with an adjustable articulator and those made with a simple articulator. Among the 36 prosthodontists, 75% often and 8% occasionally did the occlusal adjustment when giving the newly made dentures to their patients. The great majority of prosthodontists in two of top four Chinese dental hospitals, who believed most of their patients satisfied with their complete dentures, seldom used a face-bow. Occlusal adjustment may be helpful to eliminate the possible errors caused by not using a face-bow. PMID- 18422514 TI - Comparison of food mixing ability among mandibulectomy patients. AB - Many papers have been published on surgical mandibulectomy and reconstruction. However, only a few reports refer to masticatory function after prosthodontic treatment in mandibulectomy patients. The aim of this study was to investigate the masticatory function of mandibulectomy patients. Twenty-three subjects (10 males and 13 females, with an average age of 63 years) participated in this study: 11 subjects who had undergone unilateral marginal mandibulectomy, six subjects with unilateral segmental mandibulectomy with reconstruction and six subjects with hemimandibulectomy without reconstruction. Mixing Ability Index (MAI) was used to measure masticatory function on the non-defect side and on the defect side with a prosthesis installed. Comparisons were carried out among the marginal, segmental and hemimandibular groups and between the non-defect side and the defect side. Consequently, our study indicates these results. On the non defect side, a significant difference was found between the marginal and the segmental groups, and between the marginal and the hemimandibular groups. In the marginal and the segmental groups, a significant difference was found between the non-defect and the defect sides. In conclusion, our study suggests that MAI is an adequate tool to study the masticatory function in mandibulectomy patients, the masticatory function of the mandibulectomy patients is more impaired than that of the ordinary removable partial denture patients, and that surgical intervention affects the masticatory function on not only the defect side but also the non defect side in mandibulectomy patients. PMID- 18422515 TI - Leptomeningeal collateral volume flow assessed by quantitative magnetic resonance angiography in large-vessel cerebrovascular disease. AB - BACKGROUND/PURPOSE: Leptomeningeal collateral volume flow has not been previously quantified. Quantitative magnetic resonance angiography (QMRA) can determine flow in the large vessels of the intracranial circulation. METHODOLOGY: We reviewed consecutive QMRA studies performed between December 1, 2004 and August 30, 2005, for cases showing asymmetrically higher flow in a posterior cerebral artery (PCA) just distal to the origin of the posterior communicating artery ipsilateral to a hemodynamic middle cerebral artery (MCA) or internal carotid artery lesion. The mean, range, and standard deviation (SD) of the flow rate in the PCAs, MCAs, and PCA ipsilateral-contralateral difference were calculated. Ipsilateral and contralateral PCA flow rates were compared using the Student's t-test. RESULT: Sixteen studies met selection criteria. Mean age was 52 years (range 21-79) and 9 were female. MCA flow was below QMRA detection limits in 6 studies. Mean measurable ipsilateral MCA flow reduction was 84 mL/min (range 9-147, SD 51.4). Mean ipsilateral PCA flow was 118 mL/min (range 72-206, SD 38.5) and mean contralateral PCA flow was 68 mL/min (range 35-144, SD 30.5, P < .001); mean difference was 50 mL/min (range 10-93, SD 24.3). CONCLUSION: Leptomeningeal collateral flow can be assessed with QMRA and may be substantial. PMID- 18422516 TI - Qureshi grading scheme predicts subsequent volume of brain infarction following intra-arterial thrombolysis in patients with acute anterior circulation ischemic stroke. AB - BACKGROUND: The importance of the site of occlusion and the presence or absence of collaterals on initial angiography in patients with acute ischemic stroke has been recognized. Qureshi recently proposed a scheme that categorizes patients with ischemic stroke based on findings observed on initial angiography. METHODS: We determined the relationship between severity of angiographic occlusion using Qureshi grading scheme and volume of brain infarction on follow-up computed tomography in 55 patients with anterior circulation ischemic stroke who underwent intra-arterial thrombolysis. RESULTS: A strong association was observed between Qureshi grades and volume of brain infarction (F ratio 6.2, P= .0005) after adjusting for patients' age, sex, National Institutes of Health Stroke Scale (NIHSS) score, thrombolytic used, and time interval between symptom onset and angiography. The relationship persisted after further adjustment for final angiographic recanalization (F ratio 5.1, P= .001). A significant relationship between initial grades and volume of brain infarction was separately observed in both patients with or without recanalization following treatment. CONCLUSIONS: Qureshi grading scheme can be effectively used to stratify patients with anterior circulation ischemic stroke undergoing intra-arterial thrombolysis using initial angiographic findings. PMID- 18422517 TI - Vascular endothelial growth factor-165 gene therapy promotes cardiomyogenesis in reperfused myocardial infarction. AB - BACKGROUND: Vascular endothelial growth factor (VEGF)-165 promotes cardiomyogenesis in chronic myocardial ischemia and nonreperfused myocardial infarction (MI). It is unknown whether this effect is present in reperfused MI. We sought to investigate the effect of VEGF-165 gene therapy on cardiomyogenesis after reperfused MI. METHODS AND RESULTS: Twenty-four Yucatan minipigs underwent thoracotomy and a vascular clamp was placed in the left circumflex artery. Reperfusion was reestablished after 90 minutes, and VEGF-165 gene therapy or placebo was administered. A replication-deficient recombinant human adenovirus serotype 5 was used for gene transfer (Ad5-VEGF165). The same viral vector devoid of VEGF gene (Ad5-beta-galactosidase) was used as placebo. Two administration routes were tested, intramyocardial (IM) injection and circumflex intracoronary (IC) infusion. The pigs were assigned to one of the following groups: IM Ad5 VEGF165 (n = 6), IM Ad5-betaGal (n = 6), IC Ad5-VEGF165 (n = 6), and IC Ad5 betaGal (n = 6). All pigs received 5-bromo-2'-deoxyuridine (BrdU) 250 mg IV twice a week to label cells undergoing DNA replication. The hearts were explanted at 4 weeks. BrdU-labeled cardiomyocytes in the peri-infarct area were counted by a pathologist blinded to group assignment. The number of BrdU-labeled cardiomyocytes per million cells was 4-fold higher in the group receiving IM VEGF 165 (64 +/- 11.4) vs. IM placebo (16 +/- 10.6), P = 0.034. No difference in infarct size or ventricular function was observed between the groups. CONCLUSIONS: IM VEGF-165 gene therapy promotes cardiomyogenesis in reperfused MI. However, no benefit in infarct size or cardiac function was observed at 4 weeks. The origin of these cells remains unknown and needs to be determined. PMID- 18422518 TI - Long-term (up to 18 years) clinical and echocardiographic results of mitral balloon valvuloplasty in children in comparison with adult population. AB - AIMS: The purpose of this study was to assess the safety, efficacy, and long-term results (up to 18 years) of mitral balloon valvuloplasty (MBV) in children in comparison to adults. METHODS: 57 children age < or = 18 years (group A) and 474 adult patients (group B) who underwent successful MBV and were followed up for a mean 8.5 +/- 4.8 (range 1.5-18) years were analyzed. RESULTS: Patients in group A had a lower mitral echocardiographic score (echo score), 7.6 +/- 1.3 vs. 8.1 +/- 1 (P = 0.0005); smaller Doppler mitral valve area (MVA), 0.82 +/- 0.16 cm(2) vs. 0.92 +/- 0.17 cm(2) (P < 0.0001); and higher mitral valve gradient, 15.2 +/- 2.3 mmHg vs. 14.3 +/- 2.1 mmHg (P = 0.0003), than group B. Immediately after MBV, group A had larger MVA whether measured by catheter, 1.99 +/- 0.57 cm(2) vs. 1.8 +/- 0.52 cm(2) (P < 0.001), or by Doppler, 2.0 +/- 0.27 cm(2) vs. 1.97 +/- 0.28 cm(2) (P < 0.01), and similar complication rates, compared to group B. After a mean follow-up of 8.5 +/- 4.8 (range 1.5-18 years), restenosis in group A was 26% vs. 31% for group B (P = 0.41). Echo score > 8 (P = 0.046) was a predictor of restenosis in children and echo score > 8 (P < 0.0001) and previous surgery (P = 0.043) were predictors of restenosis in adults. Actuarial freedom from restenosis at 10, 15, and 18 years for groups A and B were 78%+/- 7%, 64%+/- 9%, and 18%+/- 14% and 77%+/- 2%, 43%+/- 4%, and 17%+/- 4%, respectively (P = 0.26). Event-free survival rates at 10, 15, and 18 years were 87%+/- 6%, 62%+/- 1%, and 20%+/- 2% versus 87%+/- 1%, 51%+/- 4%, and 20%+/- 5% for groups A and B, respectively (P = 0.51). Postprocedure MVA < 2.0 cm(2) (P = 0.043) and previous surgery (P = 0.03) were identified as predictors of events in children. Echo score > 8 (P < 0.0001) and prevalvuloplasty AF (P = 0.03) were identified as predictors of events in adults. CONCLUSION: MBV is safe and effective in children with rheumatic MS. It provides better immediate results than in adults and excellent long-term results that are comparable to those seen in adults. PMID- 18422519 TI - Diazepam versus fentanyl for premedication during percutaneous coronary intervention: results from the Myocardial Protection by Fentanyl during Coronary Intervention (PROFIT) Trial. AB - BACKGROUND: Sedation is a cornerstone in the premedication for percutaneous coronary intervention (PCI). Benzodiazepines and opioids are frequently used. Previous results suggest that opioids mimic the adaptation to ischemia during repeated balloon inflations and may provide direct myocardial protection in addition to their sedative effect. However, no comparative data exist. METHODS: We conducted a prospective, randomized, controlled, single-blind trial comparing diazepam and fentanyl in 276 patients undergoing elective PCI. Patients were randomized to either diazepam 5 mg sublingually or fentanyl 0.05 mg or 0.1 mg intravenously at least 5 minutes prior to the first balloon inflation. The primary end-point was the postprocedural elevation of myocardial markers of necrosis defined as an elevation of cardiac troponin T > or = 0.01 ng/ml. RESULTS: The three groups had similar baseline clinical, angiographic, and procedural characteristics, with no significant differences in lesion morphology, procedural complexity, or adjunctive medical treatment. No significant variation in the hemodynamic response to the study drugs was observed in the three groups. The rate of postprocedural troponin T elevation was 28% in the diazepam group, 20% in the fentanyl 0.05 mg group, and 30% in the fentanyl 0.1 mg group (P = 0.26). Rates of postprocedural myocardial infarction were 3%, 2%, and 2%, respectively (P = 0.84), with one case of in-hospital death in the diazepam group and no urgent TVR in the whole study population. CONCLUSION: Although providing a well-tolerated alternative to diazepam for sedation during PCI, fentanyl did not provide additional cardioprotection assessed through the postinterventional elevation of cardiac troponin T during elective coronary intervention. PMID- 18422520 TI - Observance of antiplatelet therapy after stent implantation in patients under chronic oral anticoagulant treatment. AB - PURPOSE: Patients undergoing coronary stenting must take dual antiplatelet therapy during a variable period. The combination of chronic oral anticoagulants (COA) with antiplatelet therapy has been related to an increased risk of hemorrhage. The aim of this study was to evaluate the level of the antiplatelet therapy observance in those patients and the incidence of adverse events after 1 year. METHODS: Patients with prior COA treatment with coronary lesions suitable for stenting were included. Clinical assessment was performed on admission, with follow-up at 1, 6, and 12 months. Antiplatelet and COA treatment, adverse cardiac events, and hemorrhagic episodes were registered. RESULTS: A total of 70 patients were included. Mean age was 70.5 +/- 8.7 years. The most common cause of COA was atrial fibrillation. Conventional stents were used in 40% and drug-eluting stents (DES) in 60%. Treatment at discharge was: ASA + clopidogrel + COA 64.2%, ASA + clopidogrel 25.4%, COA + clopidogrel 7.5%, and COA + ASA 3%. Observance of antiplatelet and COA therapy at 1-6-12 month follow-up after conventional stent was: COA 73.1-70.8-69.6%; ASA 92.3-75.4-65.2%; clopidogrel 92.3-62.5-43.5%. In patients receiving DES, it was: COA 76.9-78.9-80.6%, ASA 79.5-65.8-55.7%, and clopidogrel 94.9-84.2-61.1%. Dual antiplatelet therapy in patients with DES over these periods was taken in 79.5-51.4-27.8%, respectively. The incidence of adverse events was minor bleeding 11.4%, major bleeding 8.6%, myocardial infarction 4.3%, stent thrombosis 1.4%, and death 12.8%. CONCLUSIONS: There is a great variability in the treatment prescribed at discharge. Low observance with dual antiplatelet therapy has been detected in these patients, particularly after DES implantation, and they present a very high rate of complications in the follow-up. PMID- 18422522 TI - Endothelial nitric oxide synthase overexpression by neuronal cells in neurodegeneration: a link between inflammation and neuroprotection. AB - The roles of neuronal and inducible nitric oxide synthases in neurones have been extensively investigated; by contrast, the biological significance of endothelial nitric oxide synthase (eNOS) overexpression that occurs in several pathological conditions has not yet been studied. We have started addressing this issue in a cell model of neurodegeneration, i.e. human SKNBE neuroblastoma cells transfected with a mutant form of alsin, a protein causing an early-onset type of amyotrophic lateral sclerosis, ALS2. We found that eNOS, which is endogenously expressed by these cells, was activated by tumour necrosis factor-alpha (TNF-alpha), a proinflammatory cytokine that plays important roles in ALS2 and several neurodegenerative diseases. The TNF-alpha-dependent eNOS activation occurred through generation, by sphingosine-kinase-1, of sphingosine-1-phosphate, stimulation of its membrane receptors and activation of Akt, as determined using small interference RNA and dominant negative constructs specific for the enzymes and receptors. eNOS activation by TNF-alpha conferred cytoprotection from excitotoxicity and neurotoxic cues such as reactive oxygen species, endoplasmic reticulum stress, DNA damage, and mutated alsin itself. Our results suggest that overexpression of eNOS by neurones is a broad-range protective mechanism activated during damage and establish a link of pathophysiological relevance between this enzyme and inflammation accompanying neurodegenerative diseases. These findings also question the concept that high NO output in the presence of oxidative stress leads always to peroxynitrite formation contributing to neurodegeneration. PMID- 18422523 TI - Tissue engineering: strategies, stem cells and scaffolds. AB - Tissue engineering scaffolds are designed to influence the physical, chemical and biological environment surrounding a cell population. In this review we focus on our own work and introduce a range of strategies and materials used for tissue engineering, including the sources of cells suitable for tissue engineering: embryonic stem cells, bone marrow-derived mesenchymal stem cells and cord-derived mesenchymal stem cells. Furthermore, we emphasize the developments in custom scaffold design and manufacture, highlighting laser sintering, supercritical carbon dioxide processing, growth factor incorporation and zoning, plasma modification of scaffold surfaces, and novel multi-use temperature-sensitive injectable materials. PMID- 18422524 TI - Impaired meningeal development in association with apical expansion of calvarial bone osteogenesis in the Foxc1 mutant. AB - Loss of function of the mouse forkhead/winged helix transcription factor Foxc1 induces congenital hydrocephalus and impaired skull bone development due to failure of apical expansion of the bone. In this study we investigated meningeal development in the congenital hydrocephalus (ch) mouse with spontaneous loss of function mutant of Foxc1, around the period of initiation of skull bone apical expansion. In situ hybridization of Runx2 revealed active apical expansion of the frontal bone begins between embryonic day 13.5 and embryonic day 14.5 in the wild type, whereas expansion was inhibited in the mutant. Ultrastructural analysis revealed that three layers of the meninges begin to develop at E13.5 in the basolateral site of the head and subsequently progress to the apex in wild type. In ch homozygotes, although three layers were recognized at first at the basolateral site, cell morphology and structure of the layers became abnormal except for the pia mater, and arachnoidal and dural cells never differentiated in the apex. We identified meningeal markers for each layer and found that their expression was down-regulated in the mutant arachnoid and dura maters. These results suggest that there is a close association between meningeal development and the apical growth of the skull bones. PMID- 18422525 TI - Three-dimensional venous anatomy of the dermis observed using stereography. AB - Veins of the dermis have been investigated mainly by histological methods in the fields of anatomy and histology, and a large number of schemata of the veins have been depicted in a variety of textbooks. However, the schemata are usually two dimensional and it is therefore difficult to envisage the actual vasculature of the dermal veins. In this study, we performed a stereographic study of the skin of three fresh cadavers that had been injected with radio-opaque dye, which was dispersed throughout the entire body. A venous network consisting of venous polygons of various sizes existed just under the dermis or in the deep zone of the dermis, which is generally called the subdermal venous plexus. There were many small vessels towards the inside of each venous polygon, and most of them ascended, branching off stereoscopically. Those branches anastomosed with each other, and they formed the dermal and subpapillary venous plexuses. However, there was little vascular connection between dermal venous plexuses of different venous polygons. The characteristic structure of the dermal venous plexus has been considered to bring about venous congestion of the skin in various clinical situations. PMID- 18422526 TI - The relationship between intellectual disability, Indigenous status and risk of reoffending in juvenile offenders on community orders. AB - BACKGROUND: Intellectual disability (ID), age and aboriginal status have been independently implicated as risk factors for offending to varying degrees. This study examined the relationship between age, ID and the Indigenous status of juvenile offenders. It also examined the outcomes of the sample's offending in terms of court appearances and sentencing, criminogenic needs and risk of reoffending. METHOD: The sample comprised 800 juvenile offenders on community orders of whom 19% were Indigenous, who completed the New South Wales Young People on Community Order Health Survey between 2003 and 2005. Risk and criminogenic needs were evaluated using the Youth Level of Service/Case Management Inventory (Australian Adaptation) (YLS/CMI: AA). RESULTS: Those with an ID were found to have a higher risk of reoffending than those without an ID. Those with an ID were also more likely to be younger and Indigenous. For Indigenous young offenders, there was no difference between those with and without an ID in risk category allocation or number of court dates. For non Indigenous young offender, those with an ID had higher risk scores and more court dates. CONCLUSIONS: This study provided evidence that Indigenous status may play a significant role in the relationship between ID and offending in juvenile offenders on community orders. These findings have clear implications for the 'risk', 'needs' and 'responsivity' principles of offender classification for treatment. Emphasis is placed on the requirement for addressing the needs of Indigenous juvenile offenders with an ID. PMID- 18422528 TI - Women with intellectual disability at risk of adverse pregnancy and birth outcomes. AB - BACKGROUND: An increasing number of women with intellectual disability (ID) have children. Cross-sectional, clinical population data suggest that these women face an increased risk of delivering preterm and/or low birthweight babies. The aim of this study was to explore the prevalence of poor pregnancy and birth outcomes in women with ID and/or self-reported learning difficulties in an antenatal population. METHODS: A total of 878 pregnant women attending their first antenatal clinic visit were 'screened' for ID. Pregnancy and birth outcomes data were extracted from medical records post-partum. These data included pregnancy related health conditions, including pre-eclampsia and gestational diabetes, and birth outcomes, including gestational age, birthweight, Apgar score and admission to neonatal intensive care and/or special care nursery. RESULTS: A total of 57 (6.5%) pregnant women with ID and/or self-reported learning difficulties were identified. These women experienced an unusually high rate of pre-eclampsia (odds ratio = 2.85). Their children more often had low birthweights (odds ratio = 3.08), and they were more frequently admitted to neonatal intensive care or special care nursery (odds ratio = 2.51). CONCLUSION: Further research is needed to understand the reasons for the adverse findings of this study and identify potentially changeable factors contributing to adverse pregnancy and birth outcomes for women with ID and/or self-reported learning difficulties and their children. To ensure quality antenatal care, health professionals may need to consider innovations such as extended consultation times, communication aids and audio-taping consultations. PMID- 18422527 TI - 'Hungry Eyes': visual processing of food images in adults with Prader-Willi syndrome. AB - BACKGROUND: Prader-Willi syndrome (PWS) is a genetic disorder associated with intellectual disabilities, compulsivity, hyperphagia and increased risks of life threatening obesity. Food preferences in people with PWS are well documented, but research has yet to focus on other properties of food in PWS, including composition and suitability for consumption. It is also unclear how food perceptions differ across the two major genetic subtypes of PWS. METHODS: This study examined neural responses to food stimuli in 17 adults with PWS, nine with paternal deletions and eight with maternal uniparental disomy (UPD), and in nine age-matched typical controls. Visual event-related potentials (ERPs) were recorded in response to food images varying in food composition and suitability for consumption during a passive viewing paradigm. RESULTS: Group differences were observed for the N1 and P3 responses reflecting perceptual categorisation and motivational relevance respectively. The deletion group categorised food stimuli in terms of composition while the UPD group performed more similar to the controls, and focused on the suitability of food for consumption. Individual differences in N1 amplitude correlated with body mass index and scores on the Hyperphagia Questionnaire. CONCLUSION: Differences are seen in how people with PWS because of deletion or UPD perceive visual food stimuli even within the first milliseconds of stimuli exposure. Implications are discussed for in vivo food behaviours and for future ERP or neuroimaging studies on hunger, satiety and food perception in PWS. PMID- 18422529 TI - Adaptive topography of fluctuating selection in a Mendelian population. AB - An adaptive topography is derived for a large randomly mating diploid population under weak density-independent selection in a fluctuating environment. Assuming a stationary distribution of environmental states with no temporal autocorrelation, a diffusion approximation for population size and allele frequency, p, reveals that the expected change in p involves the gradient with respect to p of the stochastic intrinsic rate of increase (the density-independent long-run growth rate), r = r - sigma 2 e/2, where r is the mean Malthusian fitness in the average environment and is the environmental variance in population growth rate. The expected relative fitness of a genotype is its Malthusian fitness in the average environment minus the covariance of its fitness with population growth rate. The influence of fitness correlation between genotypes is illustrated by an analysis of the Haldane-Jayakar model of fluctuating selection on a single diallelic locus, and on two loci with additive effects on a quantitative character. PMID- 18422530 TI - Multiple paternity or multiple queens: two routes to greater intracolonial genetic diversity in the eusocial Hymenoptera. AB - Understanding the evolution of multiple mating by females (polyandry) is an important question in behavioural ecology. Most leading explanations for polyandry by social insect queens are based upon a postulated fitness benefit from increased intracolonial genetic diversity, which also arises when colonies are headed by multiple queens (polygyny). An indirect test of the genetic diversity hypotheses is therefore provided by the relationship between polyandry and polygyny across species, which should be negative if the genetic diversity hypotheses are correct. Here, we conduct a powerful comparative investigation of the relationship between polyandry and polygyny for 241 species of eusocial Hymenoptera (ants, bees and wasps). We find a clear and significant negative relationship between polyandry and polygyny after controlling for phylogeny. These results strongly suggest that fitness benefits resulting from increased intracolonial genetic diversity have played an important role in the evolution of polyandry, and possibly polygyny, in social insects. PMID- 18422531 TI - Dispersal and population structure of a New World predator, the army ant Eciton burchellii. AB - The army ant Eciton burchellii is probably the most important arthropod predator in the Neotropics, and many animal species depend upon it. Sex-biased dispersal with winged males and permanently wingless queens may render this species especially sensitive to habitat fragmentation and natural barriers, which might have severe impacts on population structure and lead to population decline. Using nuclear microsatellite markers and mitochondrial sequences, we investigated genetic differentiation in a fragmented population in the Panama Canal area. While nuclear markers showed little differentiation between subpopulations (F(ST) = 0.017), mitochondrial differentiation was maximal in some cases (Phi(ST) = 1). This suggests that, while females are not capable of crossing barriers such as large rivers, flying males are able to promote nuclear gene flow between the studied forest patches. Consistent with this interpretation, we did not find any evidence for inbreeding or genetic deterioration on Barro Colorado Island over the last 90 years since its formation. PMID- 18422532 TI - Mutualism, hybrid inviability and speciation in a tropical ant-plant. AB - Although biotic interactions are particularly intricate in the tropics, few studies have examined whether divergent adaptations to biotic interactions lead to speciation in tropical organisms. Ant-plant mutualisms are widespread in the tropics. Within Leonardoxa africana, two subspecies present contrasting defences against herbivores. Young leaves of subsp. africana are defended by mutualistic ants, whereas subsp. gracilicaulis satiates herbivores by synchronized leaf production. Subsp. africana possesses hollow internodes and many large foliar nectaries, housing and feeding ants. We detected no genetic introgression between the two subspecies in the contact zone between them. F1 hybrids were present. They were intermediate in phenotype, expressing reduced, nonfunctional but costly myrmecophilic traits. However, they suffered more herbivory than their parents. Hybrids remained small, failing to reach reproductive size, probably due to their maladapted defence phenotype. Hence, there could be a direct link between adaptation to mutualism and reproductive isolation: biotic interactions could be a driver of tropical diversity. PMID- 18422533 TI - An uncommon association: elastosis perforans serpiginosa and osteogenesis imperfecta. PMID- 18422534 TI - Naevus of Ota presenting in two generations: a mother and daughter. PMID- 18422535 TI - Improvement of pruritus and quality of life of children with atopic dermatitis and their families after joining support groups. AB - INTRODUCTION: Atopic dermatitis places a large burden on patients and their families, with greater risk of emotional disorders and behavioural problems. Preliminary evidence suggests that support groups and educational programs are helpful in reducing stress, disease and pruritus severity and improves quality of life (QoL). OBJECTIVES: To evaluate the intensity of pruritus and the QoL in children with atopic dermatitis and their families after joining support groups. Material and methods Subjects were randomly assigned to intervention or control group and completed the Children's Dermatology Life Quality Index (CDLQI) and Family Dermatitis Impact (FDI). Pruritus was evaluated by the Yosipovitch's questionnaire for pruritus. Each patient/family unit was considered as one 'patient'. Participants were divided into two different groups: one with children under 16 years and the second with patients' relatives. Each unit was accompanied during 6 months. RESULTS: Thirty-two patients and their relatives completed the questionnaires satisfactorily. After intervention, pruritus intensity was similar (P = 0.42), but the pattern of pruritus improved in the intervention group. Overall QoL for CDLQI instruments improved significantly (P < 0.01) and, when specific domains were analysed, personal relationships (P = 0.02) and leisure (P = 0.04) showed marked enhancement. FDI scores failed to demonstrate differences in the QoL of patients' relatives after treatment. CONCLUSION: The improvement on pruritus and QoL showed that atopic dermatitis patients had benefits with the attendance to support groups. We consider that these non-pharmacological approaches can be a very effective accessory tools in the management of recalcitrant forms of the disease. PMID- 18422536 TI - Herpes zoster virus associated 'sparing phenomenon': is it an innate possess of HZV or keratinocyte cytokine(s) mediated or combination? PMID- 18422537 TI - Recalcitrant acrodermatitis continua of hallopeau treated with calcitriol and tacrolimus 0.1% topical treatment. PMID- 18422538 TI - Malignant proliferating trichilemmal tumour of the scalp managed with micrographic surgery and sentinel lymph node biopsy. PMID- 18422539 TI - Anti-nuclear antibody: a potential predictor of calciphylaxis in non-dialysis patients. PMID- 18422540 TI - Pigmented purpuric dermatosis as a delayed reaction to medroxyprogesterone acetate. PMID- 18422541 TI - Allergic contact dermatitis to a temporary black harquus tattoo. PMID- 18422542 TI - Human papillomavirus 1 induced epidermoid cystic structure mimicking molluscum bodies. PMID- 18422543 TI - Multiple synchronous melanomas determined as primary by lymphatic mapping. PMID- 18422545 TI - Patients' perspective of pruritus in chronic plaque psoriasis: a questionnaire based study. AB - BACKGROUND: Pruritus in psoriasis patients has not been regarded as a major symptom. Objective To study the pattern of pruritus in chronic plaque psoriasis. METHODS: A questionnaire was sent out to 109 patients with a diagnosis of chronic plaque psoriasis, who attended our outpatient departments during the period of January 2006 to January 2007. RESULTS: Out of 109 patients, 80 patients (74%) answered the questionnaire. Pruritus was found in 80% of the patients, with an intensity of 5.2 +/- 2.6 (+/-SD) using a visual analogue scale (0-10). The frequency and intensity of pruritus were higher in women. Lower leg and scalp were reported to be the most commonly affected sites. Major aggravating factors for pruritus were stress and dryness of skin. Sun, sleep and vacation could relieve pruritus. The most common antipruritic treatments used by the patients were topical steroids, topical vitamin D, emollients and ultraviolet light therapy, whereas antihistamines were used by a small number of patients. Mood, concentration and sleep were negatively affected by pruritus. CONCLUSION: Pruritus is a common symptom in patients with chronic plaque psoriasis. PMID- 18422546 TI - Oily fish intake during pregnancy--association with lower hyperactivity but not with higher full-scale IQ in offspring. AB - BACKGROUND: Long-chain omega-3 polyunsaturated fatty acids are thought to be important for fetal neurodevelopment. Animal studies suggest that a deficiency of omega-3 fatty acids may lead to behavioural or cognitive deficits. As oily fish is a major dietary source of omega-3 fatty acids, it is possible that low intake of fish during pregnancy may have adverse effects on the developing fetal brain. METHODS: We used the Strengths and Difficulties Questionnaire and the Wechsler Abbreviated Scale of Intelligence to assess behavioural problems and intelligence in 217 nine-year-old children. The mothers of these children had participated in a study of nutrition during pregnancy during which fish intake was assessed in early and late gestation. RESULTS: Children whose mothers had eaten oily fish in early pregnancy had a reduced risk of hyperactivity compared to those whose mothers did not eat oily fish: OR .34, 95% CI .15 to .78, after adjustment for potential confounding factors. Children whose mothers had eaten fish (whether oily or non-oily) in late pregnancy had a verbal IQ that was 7.55 points higher (95% CI .75 to 14.4) than those whose mothers did not eat fish. There were, however, no significant associations between fish intake in pregnancy and other behavioural problems or full-scale and performance intelligence, after adjustment for potential confounding factors. CONCLUSIONS: Although maternal fish intake in pregnancy was associated with hyperactivity scores and verbal IQ in children, in general, how much fish women ate during pregnancy appeared to have little long term relation with neurodevelopmental outcomes in their child. PMID- 18422547 TI - Pilot study of response inhibition and error processing in the posterior medial prefrontal cortex in healthy youth. AB - BACKGROUND: Recent neuroimaging work suggests that inhibitory and error processing in healthy adults share overlapping, but functionally distinct neural circuitries within the posterior medial frontal cortex (pMFC); however, it remains unknown whether the pMFC is differentially engaged by response inhibition compared to error commission in the developing brain. Developmental neuroimaging studies of response inhibition have found pMFC activation, but the possible contribution of error-related activation during inhibitory processing has not been well studied in youth. METHOD: To examine the processing of correct response inhibition compared to errors in the developing brain, we performed functional magnetic resonance imaging scans in 11 healthy subjects, ages 8-14 years, during an antisaccade task while performance was monitored. RESULTS: Successful antisaccades activated the pre-supplementary motor area. In contrast, errors on the antisaccade task activated the dorsal anterior cingulate cortex. CONCLUSION: The findings suggest the functional sub-specialization of inhibitory and error processing within the pMFC in this pilot sample of children and adolescents. Future neuroimaging studies of developing inhibitory control should examine both between correct and error trials. PMID- 18422549 TI - Decision-making in ADHD: sensitive to frequency but blind to the magnitude of penalty? AB - BACKGROUND: Decision-making and reinforcement sensitivity were investigated in 23 children with ADHD and 20 healthy controls using a gambling paradigm. METHODS: Children were required to choose between three alternatives that carried (A) small rewards and small penalties (advantageous), (B) large rewards and increasing penalties and (C) small rewards and increasing penalties (both disadvantageous). Penalties increased either in frequency or magnitude in two independent conditions. Heart rate (HR) and skin conductance (SC) were measured to examine whether impaired decision-making was accompanied by autonomic abnormalities. RESULTS: Children with ADHD showed a maladaptive response style compared to controls by demonstrating a smaller preference for the advantageous alternative, when penalties increased in magnitude. When penalties increased in frequency, children with ADHD performed like controls. Group differences in decision-making attenuated after the task was administered twice. Compared to controls, performance of children with ADHD in the magnitude condition was accompanied by increased HR acceleration following reward. In this condition, the post-selection SC of children with ADHD was larger for advantageous than for disadvantageous alternatives, in contrast to controls who showed an opposite SC pattern. CONCLUSIONS: The current findings suggest that during decision-making, children with ADHD may be sensitive to the frequency but blind to the magnitude of penalty. PMID- 18422548 TI - Atypical development of face and greeble recognition in autism. AB - BACKGROUND: Impaired face processing is a widely documented deficit in autism. Although the origin of this deficit is unclear, several groups have suggested that a lack of perceptual expertise is contributory. We investigated whether individuals with autism develop expertise in visuoperceptual processing of faces and whether any deficiency in such processing is specific to faces, or extends to other objects, too. METHOD: Participants performed perceptual discrimination tasks, including a face inversion task and a classification-level task, which requires especially fine-grained discriminations, on three classes of stimuli: socially-laden faces, perceptually homogenous novel objects, greebles, and perceptually heterogeneous common objects. RESULTS: We found that children with autism develop typical levels of expertise for recognition of common objects. However, they evince poorer recognition for perceptually homogenous objects, including faces and, most especially, greebles. CONCLUSIONS: Documenting the atypical recognition abilities for greebles in children with autism has provided an important insight into the potential origin of the relatively poor face recognition skills. Our findings suggest that, throughout development, individuals with autism have a generalized deficit in visuoperceptual processing that may interfere with their ability to undertake configural processing, and that this, in turn, adversely impacts their recognition of within-class perceptually homogenous objects. PMID- 18422550 TI - Potassium deficiency results in accumulation of ultra-high molecular weight poly beta-hydroxybutyrate in a methane-utilizing mixed culture. AB - AIMS: To investigate the effect of various single nutrient deficiencies on poly beta-hydroxybutyrate (PHB) biosynthesis in a methane-utilizing mixed culture (dominant species Methylocystis sp. GB 25 DSM 7674). METHODS AND RESULTS: Poly beta-hydroxybutyrate accumulation experiments were performed in 7 and 70 l bioreactors and initiated by potassium, sulfur or iron deficiency. After 24 h the PHB content reached levels of 33.6%, 32.6% and 10.4% respectively. Interestingly a polymer with an ultra-high average-weight molecular weight (M(w)) of 3.1 MDa was accumulated under potassium-limited conditions. When sulfur and iron were lacking M(w) were lower by 20.6 and 41.6%. Potassium-deficiency experiments were furthermore characterized by a maximum specific PHB formation rate 0.08 g g( 1)residual biomass (R) h(-1) and a yield coefficient of 0.45 g PHB g(-1) CH(4). CONCLUSIONS: Biosynthesis of an ultra-high M(w) PHB in a methane-utilizing mixed culture can be induced by potassium deficiency. SIGNIFICANCE AND IMPACT OF THE STUDY: This study greatly extends the knowledge in the field of bacterial biopolymer formation with gaseous substrates. The special system used here combines the use of methane a low-cost substrate available from natural and renewable sources with the possibility of employing a mixed-culture in an open system for the synthesis of a high-value product. PMID- 18422551 TI - Development and antifungal evaluation of a food-grade U-type microemulsion. AB - AIMS: Food-grade microemulsions have been of increasing interest to researchers as potential delivery systems for bioactive compounds. However, food-grade microemulsions are difficult to formulate and no microemulsion has been documented for antifungal purpose. The physicochemical characterization of a food grade glycerol monolaurate (GML)/ethanol (EtOH)/Tween 80/potassium sorbate (PS)/water microemulsion system and the antifungal activities against Aspergillus niger and Penicillium italicum have been studied in this paper. METHODS AND RESULTS: The influence of EtOH and PS on oil solubilization capability was clearly reflected in the phase behaviour of U-type microemulsion systems. One dilution-stable formulation ME (GML/EtOH/Tween 80/PS/water = 3 : 3 : 3.5 : 10.5 : 16) was selected. After 4 days of incubation, ME showed 80%A. niger growth inhibition at 0.2% and 72%P. italicum growth inhibition at 0.1%, respectively, and a delay of conidiation of 2 days compared with the control. In the antifungal activities of the microemulsion, GML and PS made major contributions with similar antifungal activities at a GML/PS weight ratio of 1: 3.5. CONCLUSIONS: Food-grade dilution-stable microemulsions prepared with GML as oil phase for antifungal purpose are feasible and solubilization of a hydrotrope contributes to the formation of microemulsions and enhanced antifungal activities. SIGNIFICANCE AND IMPACT OF THE STUDY: The present report represents the first to develop a food grade microemulsion system for antifugal purpose. PMID- 18422552 TI - Antibiofilm activity of sodium bicarbonate, sodium metaperiodate and SDS combination against dental unit waterline-associated bacteria and yeast. AB - AIM: To determine the effect of sodium bicarbonate (SB), sodium metaperiodate (SMP) and sodium dodecyl sulfate (SDS) combination on biofilm formation and dispersal in dental unit waterline (DUWL)-associated bacteria and yeast. METHODS AND RESULTS: The in vitro effect of SB, SMP and SDS alone and in combination on biofilm formation and dispersal in Pseudomonas aeruginosa, Klebsiella pneumoniae, Actinomyces naeslundii, and Candida albicans was investigated using a 96-well microtitre plate biofilm assay. The combination showed a broad-spectrum inhibitory effect on growth as well as biofilm formation of both gram-negative and gram-positive bacteria, and yeast. In addition, the SB + SMP + SDS combination was significantly more effective in dispersing biofilm than the individual compounds. The combination dispersed more than 90% of P. aeruginosa biofilm whereas the commercial products, Oxygenal 6, Sterilex Ultra, and PeraSafe showed no biofilm dispersal activity. CONCLUSION: The composition comprising SB, SMP, and SDS was effective in inhibiting as well as dispersing biofilms in DUWL associated bacteria and yeast. SIGNIFICANCE AND IMPACT OF THE STUDY: This study shows that a composition comprising environmentally friendly and biologically safe compounds such as SB, SMP, and SDS has a potential application in reducing DUWL-associated acquired infections in dental clinics. PMID- 18422553 TI - Isolation of an antifungal Paenibacillus strain HT16 from locusts and purification of its medium-dependent antagonistic component. AB - AIMS: To isolate an antagonist for use in the biological control of the phytopathogenic fungus Penicillium expansum and purify the antifungal component produced by the antagonist. METHODS AND RESULTS: An antifungal strain HT16 was isolated from locusts, showing strong inhibition to Pen. expansum. Based on its in vitro effectiveness, HT16 was characterized as a strain of Paenibacillus polymyxa by phenotypic tests and 16S rDNA sequence analysis. It was found that the antifungal component HT16 secreted was only induced by Poria cocos sclerotium (PCS), and it remained active after sterilization at 121 degrees C for 15 min. The protein was purified by ammonium sulfate precipitation, heating process, and ultrafiltration using a 10 kDa cut-off membrane. The molecular weight of the purified antifungal protein, which was determined by mass spectrometry, was 4517 Da. CONCLUSIONS: A novel bacterial strain HT16 antagonistic to Pen. expansum was isolated from locusts and identified as Pae. polymyxa. The antifungal protein of 4517 Da was purified, and its production needed the inducer PCS in the fermentation medium. SIGNIFICANCE AND IMPACT OF THE STUDY: The antagonistic protein from Pae. polymyxa showed strong antifungal activity against phytopathogenic fungus Pen. expansum. This strain HT16 and the antifungal metabolite are therefore strong candidates for the biocontrol of phytopathogens in agriculture. PMID- 18422554 TI - Changes in microbial community structure, methanogenesis and rumen fermentation in response to saponin-rich fractions from different plant materials. AB - AIMS: Investigation of the effects of saponin-rich fractions on rumen fermentation, methane production and the microbial community. METHODS AND RESULTS: Saponins were extracted from Carduus, Sesbania and Knautia leaves and fenugreek seeds. Two levels of saponin-rich fractions with a substrate were incubated using the Hohenheim gas method. Methane was measured using an infrared based methane analyser and microbial communities using quantitative PCR. On addition of saponin-rich fractions, methane and short-chain fatty acid production was not affected. The protozoal counts decreased by 10-39%. Sesbania saponins decreased methanogen population by 78%. Decrease in ruminal fungal population (20 60%) and increase in Fibrobacter succinogenes (21-45%) and Ruminococcus flavefaciens (23-40%) were observed. CONCLUSIONS: The saponins evaluated possessed anti-protozoal activity; however, this activity did not lead to methane reduction. Fenugreek saponins seemed to have potential for increasing rumen efficiency. The saponins altered the microbial community towards proliferation of fibre-degrading bacteria and inhibition of fungal population. SIGNIFICANCE AND IMPACT OF THE STUDY: The uni-directional relationship between protozoal numbers and methanogenesis, as affected by saponins, is not obligatory. All saponins might not hold promise for decreasing methane production from ruminants. PMID- 18422555 TI - Characterization of rpoB mutations associated with rifampin resistance in Mycobacterium tuberculosis from eastern China. AB - AIMS: The aim of this study was to investigate the features of rpoB gene mutations associated with Rifampin (RIF) resistance in Mycobacterium tuberculosis (M. tuberculosis) in eastern China. METHODS AND RESULTS: The mutations of rpoB gene in 56 clinical isolates of M. tuberculosis resisted to one to four first line drugs (rifampin, isonicotinyl hydrazide, ethambutol and streptomycin) were analysed by polymerase chain reaction single strand conformation polymorphism analysis (PCR-SSCP) and DNA sequencing. The results of PCR-SSCP showed 52 isolates were positive (existing rpoB mutation) including 47 isolates resisted to RIF. Subsequent results of DNA sequencing showed that 54 isolates had rpoB gene mutation including 49 isolates resisted to RIF. The most frequently mutated sites were at codons 526 (73.2%), 513 (10.7%) and 531 (3.5%). CONCLUSIONS: The rpoB codon 526 was the most frequently mutated site of RIF-resistant M. tuberculosis strains in eastern China and its frequency is significantly higher (P < 0.0001) compared with that in other areas of China and in other geographic regions worldwide. SIGNIFICANCE AND IMPACT OF THE STUDY: Our results reveal that geographic variation is responsible for rpoB mutations in M. tuberculosis and the resulting information will be helpful to improve a novel rapid molecular drug resistance screening approach for MDR TB. PMID- 18422556 TI - Population structure is not a simple function of reproductive mode and larval type: insights from tropical corals. AB - 1. For a wide range of organisms, heritable variation in life-history characteristics has been shown to be strongly subject to selection, reflecting the impact that variation in characters such as genotypic diversity, duration of larval development and adaptations for dispersal can have on the fitness of offspring and the make-up of populations. Indeed, variation in life-history characteristics, especially reproduction and larval type, have often been used to predict patterns of dispersal and resultant population structures in marine invertebrates. 2. Scleractinian corals are excellent models with which to test this relationship, as they exhibit almost every possible combination of reproductive mode and larval type. Some general patterns are emerging but, contrary to expectations, genetic data suggest that while populations of broadcast spawning species may be genotypically diverse they may be heavily reliant on localized recruitment rather than widespread dispersal of larvae. 3. Here we use microsatellites to test the importance of localized recruitment by comparing the genetic structure of populations of two broadcast spawning corals with contrasting modes of reproduction and larval development; Goniastrea favulus is self-compatible, has sticky, negatively buoyant eggs and larvae and is expected to have restricted dispersal of gametes and larvae. In contrast, Platygyra daedalea is self-incompatibile, spawns positively buoyant egg-sperm bundles and has planktonic development. 4. Surprisingly, spatial-autocorrelation revealed no fine-scale clustering of similar genotypes within sites for G. favulus, but showed a non-random distribution of genotypes in P. daedalea. Both species showed similar levels of genetic subdivision among sites separated by 50 100 m (F(ST) = 0.03), suggesting that larval dispersal may be equivalent in both species. 5. Interestingly, as fragmentation has been considered rare in massive corals, our sample of 284 P. daedalea colonies included 28 replicated genotypes that were each unlikely (P < 0.05) to have been derived independently from sexual reproduction. 6. We conclude that the extreme life history of G. favulus does not produce unusually fine-scale genetic structure and subsequently, that reproductive mode and larval type may not be not good predictors of population structure or dispersal ability. PMID- 18422557 TI - Spatial population structure of a specialist leaf-mining moth. AB - 1. The spatial structure of natural populations may profoundly influence their dynamics. Depending on the frequency of movements among local populations and the consequent balance between local and regional population processes, earlier work has attempted to classify metapopulations into clear-cut categories, ranging from patchy populations to sets of remnant populations. In an alternative, dichotomous scheme, local populations have been classified as self-sustaining populations generating a surplus of individuals (sources) and those depending on immigration for persistence (sinks). 2. In this paper, we describe the spatial population structure of the leaf-mining moth Tischeria ekebladella, a specialist herbivore of the pedunculate oak Quercus robur. We relate moth dispersal to the distribution of oaks on Wattkast, a small island (5 km(2)) off the south-western coast of Finland. 3. We build a spatially realistic metapopulation model derived from assumptions concerning the behaviour of individual moths, and show that the model is able to explain part of the variation in observed patterns of occurrence and colonization. 4. While the species was always present on large trees, a considerable proportion of the local populations associated with small oaks showed extinction-recolonization dynamics. The vast majority of moth individuals occur on large trees. 5. According to model predictions, the dominance of local vs. regional processes in tree-specific moth dynamics varies drastically across the landscape. Most local populations may be defined broadly as 'sinks', as model simulations suggest that in the absence of immigration, only the largest oaks will sustain viable moth populations. Large trees in areas of high oak density will contribute most to the overall persistence of the metapopulation by acting as sources of moths colonizing other trees. 6. No single 'metapopulation type' will suffice to describe the oak-moth system. Instead, our study supports the notion that real populations are often a mix of earlier identified categories. The level to which local populations may persist after landscape modification will vary across the landscape, and sweeping classifications of metapopulations into single categories will contribute little to understanding how individual local populations contribute to the overall persistence of the system. PMID- 18422558 TI - Rainfall influences on ungulate population abundance in the Mara-Serengeti ecosystem. AB - 1. Rainfall is the prime climatic factor underpinning the dynamics of African savanna ungulates, but no study has analysed its influence on the abundance of these ungulates at monthly to multiannual time scales. 2. We report relationships between rainfall and changes in age- and sex-structured abundances of seven ungulate species monitored monthly for 15 years using vehicle ground counts in the Maasai Mara National Reserve, Kenya. 3. Abundance showed strong and curvilinear relationships with current and cumulative rainfall, with older topi, Damaliscus korrigum (Ogilby); warthog, Phacochoerus aethiopicus (Pallas); waterbuck, Kobus ellipsyprimnus (Ogilby); and impala, Aepyceros melampus (Lichtenstein) responding to longer lags than younger animals, portraying carryover effects of prior habitat conditions. 4. The abundances of newborn calves were best correlated with monthly rainfall averaged over the preceding 5-6 months for topi, waterbuck, warthog, and 2 months for the migratory zebra Equus burchelli (Gray), but with seasonal rainfall averaged over 2-5 years for giraffe, Giraffa camelopardalis (L.); impala; and kongoni, Alcelaphus busephalus (Pallas). The cumulative late wet-season rainfall was the best predictor of abundance for quarter- to full-grown animals for most species. Monthly rainfall exerted both negative and positive effects on the abundances of zebra, impala and waterbuck. Ignoring age, both sexes responded similarly to rainfall. 5. Births were strongly seasonal only for warthog and topi, but peaked between August and December for most species. Hence abundance was strongly seasonal for young topi and warthog and the migratory zebra. Pronounced seasonality in births for warthog and topi obliterated otherwise strong relationships between abundance and rainfall when both month and rainfall were included in the same model. Aggregated density produced relationships with rainfall similar to those for fully grown animals, emphasizing the necessity of demographic monitoring to reliably reveal rainfall influences on ungulate abundance in the Mara. 6. Strong relationships between abundance and rainfall suggest that rainfall underpins the dynamics of African savanna ungulates, and that changes in rainfall due to global warming may markedly alter the abundance and diversity of these mammals. Ungulates respond to rainfall fluctuations through movements, reproduction or survival, and the responses appear independent of breeding phenology and synchrony, dietary guild, or degree of water dependence. Newborns and adults have contrasting responses to rainfall. Males and females respond similarly to rainfall when age is ignored. PMID- 18422559 TI - The impact of climate fluctuation on food availability and reproductive performance of the planktivorous red-billed gull Larus novaehollandiae scopulinus. AB - 1. Using 41 years of data, we examined annual variations in the reproductive performance of the red-billed gull at the Kaikoura Peninsula, New Zealand and related these to changes in climate, oceanography and the availability of the planktonic euphausiid Nyctiphanes australis, the bird's principal food. 2. In 1994 the population began to decline, and between 1983 and 2003 it was reduced by 51%. Adult non-breeders comprised an average of 32% of the population between 1983 and 1994, but following the decline they averaged only 18%. The age at recruitment changed markedly following the population decline: 27% of 2-year-old males bred for the first time prior to the decline, whereas the corresponding figure after the decline was 38%. The proportion of females commencing to breed as a 3-year-old was not significantly different before or after the decline. 3. An increase in the availability of euphausiids increased the likelihood of breeding and the recruitment of young individuals, caused earlier laying and resulted in an increase in the condition of adults, egg volume of gulls laying two egg clutches, clutch size and fledging success. 4. The relationship between food availability and the number of pairs that bred, laying date, clutch size and fledging success was significantly different prior to and after the population decline. The underlying cause appears to be a compensatory density-dependent mechanism that reduced interspecific competition for food. 5. The relative abundance of N. australis was correlated positively with the Southern Oscillation Index (SOI) and the frequency of occurrence of NE winds. The proportion of non breeders and mean laying date were correlated negatively with the SOI, and mean egg volume of two egg clutches correlated positively with the SOI. 6. These results emphasize that availability of adult euphausiids is critical for red billed gulls. We hypothesize that high primary productivity of inshore water near Kaikoura in winter, linked to a stable water column of coastal water and upwelling of additional dissolved inorganic nutrients, increases the availability of adult euphausiids to the red-billed gull as long as the coastal water is not replaced by offshore subtropical water intrusions of warmer, low-nutrient water. PMID- 18422560 TI - Anti tumour necrosis-alpha therapy increases the number of FOXP3 regulatory T cells in children affected by Crohn's disease. AB - Crohn's disease (CD) is a chronic inflammatory disease of the gastrointestinal tract. Its pathogenesis is not completely understood, though the prevailing model is that the intestinal flora drives a strong intestinal T helper 1 (Th1)/Th17 type immune response and inflammation in the genetically susceptible host. This leads to overly aggressive T-cell responses to normal bacteria causing tissue damage. Intestinal homeostasis and maintenance of tolerance to harmless antigens in the intestine has been shown to be maintained by CD4+ CD25+ T regulatory cells (Treg) in animal models of inflammatory bowel diseases. Here we investigated whether Infliximab, a chimeric monoclonal antibody directed against tumour necrosis factor (TNF)-alpha shown to be highly effective in the treatment of CD, has any effect on mucosal CD4+ CD25+ (FOXP3+) Tregs. Colonic mucosal biopsies from children with active Crohn's disease treated in vivo with Infliximab and controls were analysed to determine FOXP3 expression by immunofluorescence and reverse transcription-polymerase chain reaction. We observed that FOXP3+ T cells were significantly reduced in mucosa of CD patients with active disease compared with controls and restored to normal following Infliximab treatment. The frequency of FOXP3+ cells and mRNA expression was significantly increased in CD mucosa from patients treated in vivo with Infliximab compared with CD patients treated with conventional therapies. In conclusion, we show that Infliximab treatment does not solely neutralize soluble TNF-alpha, but also affects activation and possibly expansion of mucosal regulatory T cells. We suggest that anti TNF-alpha immunotherapy can also restore mucosal homeostasis in Crohn's disease. PMID- 18422561 TI - Diagnosis of solid pancreatic masses by endoscopic ultrasound-guided fine-needle aspiration. AB - BACKGROUND: Endoscopic ultrasound (EUS) with fine-needle aspiration (FNA) is increasingly being used in the staging algorithm for pancreatic carcinoma. This allows for a tissue diagnosis, which was previously difficult to obtain. The aim of this study is to assess the utility of EUS-FNA in establishing the diagnosis of solid pancreatic mass lesions in an Australian population. METHODS: A retrospective review of the EUS databases of St Vincent's Hospital Melbourne and Western Hospital, Melbourne from November 2002 to May 2006 was undertaken. The focus was on patients with a solid pancreatic mass who underwent EUS-FNA. Surgical pathology or long-term follow up was used to identify false-positive or false-negative results. RESULTS: EUS was undertaken to investigate a solid pancreatic or distal common bile duct mass lesion in 155 patients. Seventy-two of these underwent EUS-guided FNA. Mean age was 68 years. A positive tissue diagnosis of malignancy could be made in 55 (76%). Nine (13%) had benign histology, with 8 (11%) having inadequate tissue obtained from FNA. A later tissue diagnosis of carcinoma was made in eight of those with either benign or inadequate histology, although in all cases there were EUS features diagnostic of malignancy, with FNA limited by technical difficulties. The overall utility of EUS-FNA showed a sensitivity of 87%, specificity 100%, positive predictive value 100%, negative predictive value 52% and overall accuracy 89%. CONCLUSION: EUS-FNA gives a high return for histological diagnosis of solid pancreatic mass lesions and should be part of the standard management algorithm for pancreatic carcinoma. PMID- 18422562 TI - Diagnostic lumbar puncture: minimizing complications. AB - Diagnostic lumbar puncture (LP) is essential to the diagnosis of central nervous system infections and subarachnoid haemorrhage. Life or limb-threatening adverse events due to the procedure are rare, but less severe complications may be common. Clinical practice in diagnostic LP is often not evidenced based. The aim of the study was to use best available published evidence to address questions on minimizing complications associated with diagnostic LP. We searched PubMed for studies in the English language using key words relevant to the complications of diagnostic LP. We emphasized randomized controlled trials and systematic reviews enrolling adult patients undergoing diagnostic LP. Uncontrolled studies and studies involving children or spinal anaesthesia were considered when no other evidence was available. There were nine prospective studies and three systematic reviews on reducing complications from LP. Recommendations on interventions to minimize complications of LP are graded based on the quality and strength of evidence. PMID- 18422563 TI - Research assessment of elder neglect and its risk factors in a hospital setting. AB - BACKGROUND: The aims of this experiment were to assess the rate of neglect in hospitalized patients and to identify risk factors for neglect. METHODS: During 2004-2006, patients aged 70 years and older hospitalized in internal departments were screened for signs of neglect. Principal caregivers were interviewed too. RESULTS: Signs of neglect were identified in 14.1% of the patients. Neglected patients differed from the non-neglected in level of education and had poorer economic status and more frailty, lower albumin level, higher incontinence problem and higher subjective caregiving burden reported by their caregivers. Mean risk scores, based on patients' and caregivers' risk indicators, were higher in the neglect group. CONCLUSION: Health, functional and psychosocial factors correlated to the presence of neglect. These factors should be used as warning signs to draw attention to professional help. PMID- 18422564 TI - Utilization of allopurinol in the Australian community. AB - BACKGROUND: International data suggest that suboptimal use of allopurinol is common. Allopurinol dose should be lower in renal impairment, but higher when gout is not controlled. The aim of the study was to examine trends in the usage of allopurinol in the Australian community. METHODS: Community dispensing data on the urate-lowering drugs allopurinol and probenecid were obtained from databases kept by Medicare Australia and the Drug Utilization Sub-Committee, for January 1992 to December 2005. RESULTS: Allopurinol comprised 98.4% of all prescriptions for urate-lowering drugs dispensed during 2005. Most prescriptions were for allopurinol 300 mg, but there was a steady shift towards use of allopurinol 100 mg in all states and territories over the period of the study. There were marked variations in prescribing rates across the country. New South Wales had the highest rate of subsidized prescribing for allopurinol 300 mg (39.3 per 1000 population). Tasmania had the highest rate for allopurinol 100 mg (14.3 per 1000 population), which coincided with an educational programme to decrease allopurinol dose in patients with renal impairment. Prescribing rates in the Northern Territory were substantially lower than all other regions, at 10.8 and 3.3 prescriptions per 1000 population for allopurinol 300 and 100 mg, respectively. CONCLUSION: The increased uptake of allopurinol 100 mg suggests greater adherence to dosing guidelines and that there is value in educational programmes to optimize drug usage. Variability in utilization rates across regions indicates the need for research on factors responsible. Precise understanding of dosing trends requires access to deidentified, individual dosing data. PMID- 18422565 TI - Tumour necrosis factor inhibitors in ankylosing spondylitis. AB - Ankylosing spondylitis (AS) is the most common form of spondyloarthropathy. Non steroidal anti-inflammatory medications and exercise are used to manage the chronic inflammatory spinal pain and stiffness. Up to 20% of patients have a peripheral inflammatory arthritis, which is treated with standard disease modifying anti-rheumatic drugs especially sulfasalazine and methotrexate. Patients may also have extra-articular manifestations, such as anterior uveitis, psoriasiform skin lesions and inflammatory bowel disease. Anti-tumour necrosis (TNF) therapy has been used with great success in rheumatoid arthritis. There are now good data of the efficacy of anti-TNF therapies in the short and medium terms in AS. Etanercept, infliximab and adalimumab have been shown in randomized placebo-controlled trials of short duration to significantly reduce disease activity, including pain and stiffness as well as improving function, spinal movement and quality of life. It is hoped that long-term therapy will prevent radiologic progression and ankylosis and studies of long-term efficacy are awaited. Anti-TNF therapies are generally well tolerated in AS. It is important to screen for latent tuberculosis before the commencement of anti-TNF therapy. The side-effect profile of anti-TNF therapies in AS does not appear different from that in rheumatoid arthritis. Currently, treatment with anti-TNF therapy in AS is indicated in established disease with radiographic damage. There is evidence that response to therapy is greater in patients with earlier disease and less damage. Future developments may see this therapy extended to patients with pre-radiographic AS. PMID- 18422566 TI - Thrombotic thrombocytopenic purpura is associated with a high relapse rate after plasma exchange: a single-centre experience. AB - BACKGROUND: Thrombotic thrombocytopenic purpura (TTP) is a rare condition characterized by microangiopathic haemolytic anaemia, thrombocytopenia, renal and/or neurological dysfunction secondary to microvascular or macrovascular thrombosis. Despite advances in treatment, TTP remains a serious condition with significant morbidity and mortality. METHODS: We undertook an audit of patients with TTP over 14 years to assess remission, relapse, survival and factors predictive of outcome using current therapy based on plasma exchange with fresh frozen plasma. RESULTS: Forty patients were identified between January 1992 and December 2005. Thirty-one (82%) achieved complete response (CR) to therapy using plasma exchange with fresh-frozen plasma (median 11 exchanges) and steroids. Twelve (37%) relapsed a median of 14 days following cessation of therapy, with multiple relapses occurring in two patients. TTP-related death occurred in four patients during their initial presentation and in two during subsequent relapse. Four patients were only partially responsive to first-line therapy. The absence of neurological features at presentation was the only factor predicting a sustained CR to first-line therapy (P = 0.027, log-rank analysis). The mean duration of inpatient treatment was 18 days (range 4-38 days) with 30% of patients requiring intensive care admission. Thirty-four per cent of patients acquired central venous line infection, with a median of two episodes of line sepsis per patient. CONCLUSION: Our results indicate the need for better treatments to reduce the high early relapse rate and significant mortality associated with current therapy. PMID- 18422567 TI - Prevalence of sicca symptoms in a South Australian cohort with systemic sclerosis. AB - BACKGROUND: The presence of sicca symptoms is a frequent finding in patients with systemic sclerosis (SSc). The aim of this study was to examine the prevalence of sicca symptoms in a South Australian cohort of SSc patients and correlate this to a number of parameters, including autoantibody status, use of anticholinergic medication, age and the presence of functional anti-muscarinic-3 receptor (M3R) blocking antibodies. METHODS: A screening questionnaire was sent out to all patients on the South Australian Scleroderma Register from the years 1998-2006 to determine the prevalence of sicca symptoms. A subset of patients on the register had ocular sicca symptoms tested by use of Schirmer's strips to validate the accuracy of the questionnaire. Eight patients were tested for anti-M3R-blocking antibodies using a functional physiological assay. RESULTS: One hundred and ninety-three SSc patients took part in this study. Sicca symptoms were present in 59% of patients with the limited form of SSc, compared with 49% of patients with the diffuse form and 40% of patients with the overlap syndrome. The use of anticholinergic medication or thyroxine was associated with higher sicca scores in SSc patients. SS-A and SS-B autoantibodies (seen in Sjogren's syndrome) were detected in eight patients in this study. The detection of anti-M3R-blocking antibodies correlated well to presence of sicca. CONCLUSION: This study confirmed that sicca symptoms are found in a high proportion of patients with SSc, especially those with the limited variant. Further testing of larger numbers of SSc patients with sicca for anti-M3R-blocking antibodies will be needed before more definitive conclusions can be drawn. Physicians should be made aware that sicca symptoms are a frequent cause of morbidity for SSc patients*. PMID- 18422568 TI - How do patients with parkinsonism present? A clinicopathological study. AB - BACKGROUND: The early clinical features of neurodegenerative parkinsonism can be subtle and often coexist with autonomic, sensory and psychic symptoms, making accurate early diagnosis challenging. METHODS: We retrospectively assessed the initial, clinical presentation and referral patterns of 494 patients with a pathological diagnosis of parkinsonism (344 with Parkinson's disease (PD), 89 with progressive supranuclear palsy (PSP) and 61 with multiple system atrophy (MSA)) archived at the Queen Square Brain Bank, London. RESULTS: Forty-four per cent of all patients were initially referred to a neurologist. Of those with PD, 28% were referred to a general physician, and approximately 5% each to orthopaedic surgeons, urologists, psychiatrists and rheumatologists. Pain was common in those not initially referred to neurologists and most lacked early tremor, rigidity and bradykinesia. More PSP patients were initially referred to ophthalmologists than in PD (9 vs 1%, chi(2)P < 0.001) and more MSA patients were referred to a urologist or gynaecologist than in PD (21 vs 5%, chi(2)P < 0.001). In PD a frozen shoulder, degenerative spine disease and benign prostatic hypertrophy were the most common early diagnoses. In PSP, the most common misdiagnosis was PD, followed by vascular parkinsonism and degenerative spinal disease. This was similar in MSA, but bladder outlet obstruction and idiopathic vocal cord palsy were other initial diagnoses. CONCLUSION: The early clinical manifestations of PD, PSP and MSA are protean and often non-specific, leading to diverse specialist referrals. When diagnoses, such as frozen shoulder and cervical spondylosis, benign prostatic hypertrophy or unstable bladder and depression are made, specialists should consider the possibility of early parkinsonism and look carefully for additional subtle motor abnormalities. PMID- 18422569 TI - An experimental study on effects of increased ventilation flow on students' perception of indoor environment in computer classrooms. AB - The effects of ventilation in computer classrooms were studied with university students (n = 355) in a blinded study, 31% were women and 3.8% had asthma. Two classrooms had a higher air exchange (4.1-5.2 ac/h); two others had a lower air exchange (2.3-2.6 ac/h). After 1 week, ventilation conditions were shifted. The students reported environmental perceptions during the last hour. Room temperature, RH, CO2, PM10 and ultra-fine particles were measured simultaneously. Mean CO2 was 1185 ppm at lower and 922 ppm at higher air exchange. Mean temperature was 23.2 degrees C at lower and 22.1 degrees C at higher air exchange. After mutual adjustment (temperature, RH, CO2, air exchange), measured temperature was associated with a perception of higher temperature (P < 0.001), lower air movement (P < 0.001), and poorer air quality (P < 0.001). Higher air exchange was associated with a perception of lower temperature (P < 0.001), higher air movement (P = 0.001), and better air quality (P < 0.001). In the longitudinal analysis (n = 83), increased air exchange caused a perception of lower temperature (P = 0.002), higher air movement (P < 0.001), better air quality (P = 0.001), and less odor (P = 0.02). In conclusion, computer classrooms have CO2 levels above 1000 ppm and temperatures above 22 degrees C. Increased ventilation from 7 l/s per person to 10-13 l/s per person can improve thermal comfort and air quality. PRACTICAL IMPLICATIONS: Computer classrooms are crowded indoor environments with a high thermal load from both students and computer equipment. It is important to control room temperature either by air conditioning, sun shields, or sufficiently high ventilation flow. A high ventilation flow is also crucial to achieving good perceived air quality. Personal ventilation flow should be at least 10 l/s. Possible loss of learning ability due to poor indoor air quality in university buildings deserves more attention. PMID- 18422570 TI - Measuring the exposure of infants and children to indoor air pollution from biomass fuels in The Gambia. AB - Indoor air pollution (IAP) from biomass fuels contains high concentrations of health damaging pollutants and is associated with an increased risk of childhood pneumonia. We aimed to design an exposure measurement component for a matched case-control study of IAP as a risk factor for pneumonia and severe pneumonia in infants and children in The Gambia. We conducted co-located simultaneous area measurement of carbon monoxide (CO) and particles with aerodynamic diameter <2.5 microm (PM(2.5)) in 13 households for 48 h each. CO was measured using a passive integrated monitor and PM(2.5) using a continuous monitor. In three of the 13 households, we also measured continuous PM(2.5) concentration for 2 weeks in the cooking, sleeping, and playing areas. We used gravimetric PM(2.5) samples as the reference to correct the continuous PM(2.5) for instrument measurement error. Forty-eight hour CO and PM(2.5) concentrations in the cooking area had a correlation coefficient of 0.80. Average 48-h CO and PM(2.5) concentrations in the cooking area were 3.8 +/- 3.9 ppm and 361 +/- 312 microg/m3, respectively. The average 48-h CO exposure was 1.5 +/- 1.6 ppm for children and 2.4 +/- 1.9 ppm for mothers. PM(2.5) exposure was an estimated 219 microg/m3 for children and 275 microg/m3 for their mothers. The continuous PM(2.5) concentration had peaks in all households representing the morning, midday, and evening cooking periods, with the largest peak corresponding to midday. The results are used to provide specific recommendations for measuring the exposure of infants and children in an epidemiological study. PRACTICAL IMPLICATIONS: Measuring personal particulate matter (PM) exposure of young children in epidemiological studies is hindered by the absence of small personal monitors. Simultaneous measurement of PM and carbon monoxide suggests that a combination of methods may be needed for measuring children's PM exposure in areas where household biomass combustion is the primary source of indoor air pollution. Children's PM exposure in biomass burning homes in The Gambia is substantially higher than concentrations in the world's most polluted cities. PMID- 18422571 TI - Health-related quality of life in Japanese men with localized prostate cancer: assessment with the SF-8. AB - OBJECTIVES: To evaluate health related quality of life (HRQOL) using the Medical Outcomes Study 8-items Short Form Health Survey (SF-8) questionnaire in Japanese patients with early prostate cancer. METHODS: A cross-sectional analysis was done in 457 patients with prostate cancer treated with radical prostatectomy, external beam radiotherapy, brachytherapy, androgen deprivation therapy, and watchful waiting or a combination these therapies. General HRQOL was measured using the Japanese version of the SF-8 questionnaire and disease-specific HRQOL was assessed using the Japanese version of the Extended Prostate Cancer Index Composite. RESULTS: The external beam radiotherapy group reported significantly lower values for the physical health component summary score (PCS) in comparison to the radical prostatectomy and brachytherapy groups (P < 0.05). In the analysis of both the PCS and the mental health component summary score (MCS) over time after treatment, higher scores with time were found in the radical prostatectomy group. No significant change over time after androgen deprivation therapy in the PCS was found. In contrast, the MCS was found to deteriorate in the early period, showing a significant increase over time. CONCLUSIONS: SF-8 in combination with the Extended Prostate Cancer Index Composite has shown to be a helpful tool in the HRQOL assessment of Japanese patients treated for localized prostate cancer. PMID- 18422572 TI - Detrusor contractility and overactive bladder in patients with cerebrovascular accident. AB - OBJECTIVES: A retrospective analysis of urodynamic studies was conducted to investigate detrusor contractility following cerebrovascular accident. METHODS: Fifty-seven patients (34 men and 23 women) who were rehabilitated following cerebrovascular accident were included in the analysis. Subjects with maximum contractile power values less than 10 W/m(2) in men and 8 W/m(2) in women, in the absence of bladder outlet obstruction, were defined as having detrusor underactivity. RESULTS: A significantly lower maximum value of detrusor contractile power and a larger residual urine volume, particularly more prevalent in women than in men, appeared unexpectedly after cerebrovascular accident. Patients with underactivity had significantly lower maximum and average flow rates, lower contractile power, and an increased postvoid residual relative to their normal counterparts. The prevalence of underactivity was 35% in the men and 43% in the women. Furthermore, 67% of the men and 80% of the women with underactivity had overactive bladder symptoms. On the other hand, detrusor underactivity was observed in 8 (35%) of 23 overactive bladder patients in the men and 8 (42%) of 19 in the women. CONCLUSIONS: Post-cerebrovascular accident patients presented various patterns of detrusor contractility. Specific attention to the possible occurrence of detrusor underactivity in these patients is essential for the successful management of overactive bladder syndrome. PMID- 18422573 TI - A novel quantitative method for measuring obstruction in the upper urinary tract: the 'obstruction coefficient'. AB - OBJECTIVES: To define an exact pressure-flow correlation in the upper urinary tract using an improved measurement method, to quantitatively characterize the degree of postrenal obstruction and to find a simple way of calculating it in everyday urological practice. METHODS: The data of 112 cases were included in the analysis. The dynamic method of a multistep, constant pressure perfusion study was used to precisely measure a wide range of pressure-flow dependences. Values of established parameters measuring the degree of obstruction were compared: the intrapelvic pressure, the ureteral opening pressure and the newly introduced 'obstruction coefficient'. RESULTS: Pressure-flow relations can be best presented by a parabolic curve described by the simple formula Y = AX(2) + B. Depending on the degree of obstruction, the shape of this curve can be characterized by a single number, that we defined as the 'obstruction coefficient'. Computer-based evaluation software for the easy calculation of this coefficient is presented here and freely available on demand. The Whitaker-test, the ureteral opening pressure, and the 'obstruction coefficient' showed significant correlation proving that the latter was clinically applicable in measuring the degree of obstruction. CONCLUSION: Calculation of the 'obstruction coefficient' enables us to exactly define the degree of upper urinary tract obstruction and to safely monitor for a long period conditions inhibiting ureteric passage. PMID- 18422574 TI - Prediction of extraprostatic extension by prostate specific antigen velocity, endorectal MRI, and biopsy Gleason score in clinically localized prostate cancer. AB - OBJECTIVES: To investigate the clinical value of prostate specific antigen velocity (PSAV) in predicting the extraprostatic extension of clinically localized prostate cancer. METHODS: One hundred and three patients who underwent radical prostatectomy for clinically localized prostate cancer were included in the analysis. The correlation between preoperative parameters, including PSA based parameters, clinical stage, and histological biopsy findings, and the pathological findings were analyzed. Logistic regression analysis was performed to identify a significant set of independent predictors for the local extent of the disease. RESULTS: Sixty-four (60.2%) patients had organ confined prostate cancer and 39 (39.8%) patients had extraprostatic cancer. The biopsy Gleason score, PSA, PSA density, PSA density of the transition zone, and PSAV were significantly higher in the patients with extraprostatic cancer than in those with organ confined cancer. Multivariate logistic regression analysis indicated that the biopsy Gleason score, endorectal magnetic resonance imaging findings, and PSAV were significant predictors of extraprostatic cancer (P < 0.01). Probability curves for extraprostatic cancer were generated using these three preoperative parameters. CONCLUSIONS: The combination of PSAV, endorectal magnetic resonance imaging findings, and biopsy Gleason score can provide additional information for selecting appropriate candidates for radical prostatectomy. PMID- 18422575 TI - Controversies in perioperative management and antimicrobial prophylaxis in urologic surgery. AB - The Japanese Urological Association (JUA) recently published guidelines for the prevention of perioperative urologic infections. Although the general remarks in the JUA guidelines are almost similar to those in guidelines previously published by the Centers for Disease Control and Prevention (CDC) and in the European Association of Urology (EAU) guidelines, their differences leave several questions that need to be answered. To clarify agreements and differences in guidelines for perioperative management in urologic interventions for development of more optimal guidelines, reports and reviews previously published were overlooked and discussed. In terms of surgical site infections (SSI) in urologic surgery, consensus for open and endoscopic-instrumental procedures is still somewhat controversial, while a consensus has not yet emerged for its use in laparoscopic procedures. Further research is required to determine what is an optimal prophylactic protocol to effectively prevent both SSI and remote infections (RI). PMID- 18422576 TI - Perioperative antibiotics in radical cystectomy with ileal conduit urinary diversion: efficacy and risk of antimicrobial prophylaxis on the operation day alone. AB - OBJECTIVE: To determine the optimum schedule for perioperative antimicrobial prophylaxis (AMP) for bladder cancer patients submitted to radical cystectomy with ileal conduit urinary diversion. METHODS: We studied 77 consecutive bladder cancer patients who underwent radical cystectomy with ileal conduit. The 1-day group (n = 33) received pre-, intra- and postoperative administrations of 2 g of piperacillin on the operation day alone; the 3-day group (n = 44) received antibiotics for 3 days or more (same schedule as the 1-day group on the operation day and every 12 h thereafter). The study was designed and postoperative complications including surgical-site infection (SSI) were defined according to the modified Centers for Disease Control and Prevention criteria. RESULTS: No significant differences were found between the 1-day group and 3-day group in terms of total SSI (18.1% vs 20.5%), superficial incisional SSI (12.1% vs 13.6%), deep incisional SSI (12.1% vs 13.6%), space SSI (12.1% vs 11.4%), postoperative ileus (18.2% vs 11.4%), febrile urinary tract infections (15.2% vs 15.9%) or pneumonia (3.0% vs 4.3%), respectively. In both groups, disease stage and patients' underlying conditions such as diabetes did not have an influence on the incidence of postoperative complications. CONCLUSION: One-day AMP had equivalent efficacy to that of the standard prophylaxis protocol for preventing septic complications following radical cystectomy with ileal conduit. This finding supports the hypothesis that delivery of antibiotics on the operation day is critical in this setting. PMID- 18422577 TI - Genetic variants of the XRCC7 gene involved in DNA repair and risk of human bladder cancer. AB - OBJECTIVE: To investigate the association between the polymorphisms of the KU70 and X-ray repair cross complementing group 7 (XRCC7) genes and the risk of bladder cancer. METHODS: This hospital-based case-control study included 213 patients with newly diagnosed bladder transitional cell carcinoma and 235 cancer free controls frequency-matched by age and sex. Two polymorphisms, KU70 and XRCC7, using a method involving polymerase chain reaction-restriction fragment length polymorphism were genotyped. RESULTS: The risk of bladder cancer decreased in a dose-response manner as the number of XRCC76721G alleles increased (adjusted odds ratio [OR] = 0.70, 95% confident interval [CI] = 0.47-1.03 for 6721GT and OR = 0.31, 95% CI = 0.10-0.99 for 6721GG; P(trend) = 0.013). However, when we used 6721 (GT + GG) as the reference, we found a statistically significant increased risk of bladder cancer associated with the 6721TT genotype (OR = 1.53, 95% CI = 1.04-2.25). In the stratification analysis, this increased risk was more pronounced among subgroups of patients aged >65 years (OR = 2.27; 95% CI = 1.25 4.10) and ever smokers (OR = 2.06, 95% CI = 1.15-3.68). Furthermore, we observed a 3.24-fold increased risk (95% CI = 1.35-7.78) for smokers aged >65 years carrying 6721TT genotype compared with those carrying the 6721 (GG + GT) genotype. However, the KU70-61C > G polymorphism was not associated with a significantly increased risk of bladder cancer. CONCLUSIONS: The XRCC7 but not the KU70 polymorphism appears to be involved in the etiology of human bladder cancer. Larger studies with more detailed data on environmental exposure are needed to verify these initial findings. PMID- 18422578 TI - Long-term outcome of secondary endopyelotomy after failed primary intervention for ureteropelvic junction obstruction. AB - OBJECTIVE: To evaluate the long-term outcome of secondary endopyelotomy after failed primary intervention for uretero-pelvic junction (UPJ) obstruction and to assess the effect of preoperative parameters on treatment outcome. METHODS: Twenty patients (13 men, seven women; mean age 30.7 years) who underwent secondary endopyelotomy after the failure of a primary intervention for the treatment of congenital UPJ obstruction were included in this retrospective analysis. Mean interval from primary treatment to secondary endopyelotomy was 27.2 months (range 3-123 months). The diagnosis of failure of the primary treatment was based on symptoms and the results of imaging studies. Treatment success was defined as symptomatic relief with either stable or improved renal function and improved wash-out shown on diuretic renogram or excretory urography. RESULTS: Mean follow-up was 47.2 months (range 6.2-138.8 months). Success rates were as follows: overall, 70%; after primary dismembered pyeloplasty, 66.7%; after primary endopyelotomy, 57.1%; after primary balloon dilatation, 100%. Kaplan-Meier estimates of success were 64.4% at 5 years. Six patients in whom the procedure failed at a mean of 13.8 months (range 4-33 months) were treated with open pyeloplasty (four patients), simple nephrectomy (one), and a repeat endopyelotomy (one). Grade 4 hydronephrosis and significant obstruction occurred more often in the failure group. CONCLUSIONS: Endopyelotomy is an acceptable minimally invasive secondary treatment option for UPJ obstruction. Preoperative severe hydronephrosis and the presence of a significant obstruction seem to be risk factors for the failure of a secondary endopyelotomy. PMID- 18422580 TI - Laparoscopic decortication of symptomatic renal cysts: experience from a referral center in Iran. AB - OBJECTIVE: To present our experience with laparoscopic management of symptomatic simple renal cysts. METHODS: From April 2004 to November 2006, 21 patients (10 men; 11 women) underwent laparoscopic decortication for simple renal cysts at our department and were included in the analysis. All procedures were carried out by one surgeon using a transperitoneal approach. Patients underwent radiological follow-up with computerized tomography and/or ultrasonography. Procedural success was defined as no recurrence of the cyst and complete pain relief. Symptomatic success was defined as a significant pain decrease. RESULTS: All 21 procedures were completed laparoscopically, without major complications or conversion to open surgery. Estimated mean blood loss during surgery was about 50 mL. Patients were hospitalized for a mean of 1.9 +/- 1.1 (range: 1-5) days. Age of the patients and size and location of the cysts, had no relationship with the duration of operation as well as the length of hospital stay (P > 0.05). Patients who experienced complete pain relief had significantly larger cyst sizes compared with patients with a partial pain decrease (7.3 +/- 1.1 vs 9.1 +/- 2.0, respectively; P = 0.023, F = 0.606). All patients had negative cytological and pathological findings for malignancy or any other abnormalities. At 16.6 months of mean follow up, none of the patients reported symptomatic and/or radiologic failure. CONCLUSIONS: Laparoscopic transperitoneal decortication represents an effective and safe treatment option in the management of symptomatic renal cysts. PMID- 18422579 TI - Low dose tamsulosin for stone expulsion after extracorporeal shock wave lithotripsy: efficacy in Japanese male patients with ureteral stone. AB - OBJECTIVE: We evaluated the efficacy of low dose tamsulosin after extracorporeal shock wave lithotripsy (ESWL) in Japanese male patients with ureteral stone. METHODS: One hundred and two Japanese male patients with ureteral stones who underwent ESWL were randomly divided into three groups. Group A (38 patients) was given tamsulosin (0.2 mg/day); group B (30 patients) was given c horeito, a herbal medicine (7.5 g/day); and group C (34 patients) received no medication. Stone clearance was assessed at 1, 7, 14, and 28 days after ESWL using plain abdominal radiography and abdominal ultrasonography. After 28 days, stone delivery was checked every 2 weeks. RESULTS: The stone-free rate was 84.21%, 90%, and 88.24% for groups A, B, and C, respectively (P = 0.3425). The mean expulsion time was 15.66 +/- 6.14 days in group A, 27.74 +/- 25.36 days in group B, and 35.47 +/- 53.70 days in group C. The expulsion time of group A was significantly shorter than that of groups B (P = 0.0116) and C (P = 0.0424). CONCLUSIONS: The addition of tamsulosin to conservative treatment appeared to be effective in shortening the stone expulsion time. PMID- 18422581 TI - Usefulness of single photon emission computed tomography imaging in the detection of lumbar vertebral metastases from prostate cancer. AB - OBJECTIVE: To determine whether single photon emission computed tomography (SPECT) is useful in the detection of prostate cancer bone metastases in the lumbar vertebrae. METHODS: Thirty-nine patients (12 with benign prostatic hyperplasia, 27 with prostate cancer) were considered and submitted to bone SPECT. All of them had increased uptake in lumbar vertebrae on bone scintigraphy. In those with prostate cancer, definitive diagnosis of bone metastases was established by magnetic resonance imaging (MRI). SPECT axial images were classified into five accumulation patterns: mosaic, large hot, diffuse, peripheral, and articular (or pediculate). Sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of bone SPECT were calculated. RESULTS: Overall, 116 vertebral lesions (49 metastatic, 67 degenerative) were studied. Mosaic, large hot and diffuse patterns were more frequently associated with metastatic lesions (84.2%, 70.3%, and 63.1% of the cases, respectively). On the other hand, peripheral and articular (or pediculate) patterns were mostly ascribed to degenerative lesions (100% and 87.5% of the cases, respectively). Sensitivity, specificity, PPV and NPV of bone SPECT were 95.9% (47/49), 73.1% (49/67), 72.3% (47/65), and 96.1% (49/51), respectively. CONCLUSIONS: Bone SPECT provides better accuracy than bone scintigraphy in differential diagnosis of lumbar vertebral lesions from prostate cancer. PMID- 18422582 TI - The technical quality of nonsurgical root canal treatment performed by a selected cohort of Australian endodontists. AB - AIM: To investigate the technical quality of nonsurgical root canal treatment performed by endodontists in Melbourne, Australia. METHODOLOGY: Clinical and radiographic records of 100 sequential nonsurgical patients were obtained from each of six endodontists working in private practice. The following variables were analysed: proximity of root filling to radiographic apex; homogeneity and radiodensity of root filling; lateral adaptation of the root filling to the canal walls; taper; extrusion of material; small, appropriate or excessive apical enlargement; presence of lateral canals; transportation; procedural errors. The radiographs were assessed by three independent evaluators. Exploratory data analysis was undertaken using simple frequencies and cross-tabulations. A generalised linear mixed model (GLMM) was used for the formal statistical modelling. RESULTS: Of the 1351 canals that were examined, 91.7% were filled within 2 mm of the radiographic apex and 74% were within 1 mm. Homogeneity and adequate density were found along the entire length of the canal in 86.1% and 88.6% of cases respectively. Lateral adaptation was adequate in 95.6% of cases and the taper was 'smooth and continuous' in 83.8% of roots. No and/or small extrusion of sealer was noted in 98.3% of cases. Apical enlargement was 'appropriate' in 85% of roots. Both transportation (1.1%) and procedural errors (1.3%) were rare occurrences. CONCLUSIONS: The technical quality of root fillings performed by endodontists in Melbourne, Australia complied with current guidelines in 77.4%-91.0% of roots. All variables examined confirmed high levels of technical proficiency. There were very few instances of canal transportation and/or procedural errors. PMID- 18422583 TI - Microbiological evaluation of primary endodontic infections in teeth with and without sinus tract. AB - AIM: To examine the microbiological status of primary endodontic infections in teeth with and without a sinus tract. METHODOLOGY: Samples were collected by means of a size 15 H-type file and two sterile paper points from 30 cases of primary endodontic infections with (n = 15) or without (n = 15) a sinus tract. The presence of 40 bacterial species was determined by the checkerboard DNA-DNA hybridization method. RESULTS: The species found at the highest levels and prevalence were Fusobacterium nucleatum sp. vincentii, Porphyromonas gingivalis, Veillonella parvula, Enterococcus faecalis, Campylobacter gracilis and Neisseria mucosa. Total bacterial counts were similar between teeth with (44 x 10(5)) and without (50 x 10(5)) a sinus tract (t-test: P > 0.05). E. faecalis, Streptococcus anginosus, Capnocytophaga sputigena and Capnocytophaga gingivalis had significantly higher counts in the absence of sinus tract (Mann-Whitney test, P < 0.05). Higher levels of P. gingivalis and Fusobacterium nucleatum sp. nucleatum were observed in cases with a sinus tract. Leptotrichia buccalis (OR = 1.83; CI 95%) and Porphyromonas endodontalis (OR = 2.15; CI 95%) were associated with an increased chance of subjects having a sinus tract. CONCLUSIONS: Primary endodontic infections were associated with a large variety of bacterial species. Specific differences between the composition of the microbiota of primary root canal infections were observed in cases with or without a sinus tract. PMID- 18422584 TI - Effects of post system and length on the strain and fracture resistance of root filled bovine teeth. AB - AIM: To investigate ex vivo the effects of different post systems and lengths on the strain and fracture resistance of root-filled teeth. METHODOLOGY: One hundred and thirty-five bovine incisors were sectioned 15 mm from their apices, root filled and divided into three groups (n = 45): fibreglass post; prefabricated steel post; cast post and core. Each group was divided into three subgroups (n = 15) according to the post length: 5.0 mm; 7.5 mm; 10.0 mm. All teeth were restored with metal crowns. For strain-gauge measurement, two strain gauges per sample were used. The fracture resistance (N) was measured and the data were analysed with two-way analyses of variance, followed by the Tukey's HSD test (alpha = 05). RESULTS: For all posts, decreased lengths resulted in increased microstrain values. However, the fibreglass posts were associated with lower increases when compared with cast post and cores and prefabricated steel posts, which showed microstrain values two times higher when the post length was 5.0 mm. The two-way analyses of fracture resistance values revealed that post length was statistically significant for the metal posts and not significant for the fibreglass post. The fracture mode analysis indicated that all groups tended to demonstrate root fractures in groups restored with metal posts and resin core fractures in groups restored with fibreglass posts. CONCLUSIONS: The cast post and core when the length was 10.0 mm had the highest fracture resistance; however, the fibreglass post was effective with the three post lengths, with higher fracture resistance than metal posts when the length was 5.0 mm. PMID- 18422585 TI - Ex vivo evaluation of the effectiveness of bleaching agents on the shade alteration of blood-stained teeth. AB - AIM: To evaluate ex vivo effectiveness of the three formulations of bleaching materials for intracoronal bleaching of root filled teeth using the walking bleach technique. METHODOLOGY: Extracted premolar teeth were stained artificially with human blood. After biomechanical preparation, the root canals were filled and a 3-mm thick intermediate base of zinc phosphate cement was placed at the level of the cementoenamel junction. The teeth were divided into four groups (n = 12): C (control, without bleaching material), A1 (sodium perborate + distilled water), A2 (sodium perborate + 10% carbamide peroxide) and A3 (sodium perborate + 35% carbamide peroxide). The bleaching materials were changed at 7 and 14 days. Evaluation of shade was undertaken with aid of the VITA Easyshadetrade mark (DeltaE*ab) and was performed after tooth staining and at 7, 14 and 21 days after bleaching, based on the CIELAB system. Data were analysed by anova for repeated measurements, Tukey and Dunnett tests (alpha = 0.05). RESULTS: The Tukey test revealed that group A1 (10.58 +/- 4.83 DeltaE*ab) was statistically different from the others (A2, 19.57 +/- 4.72 DeltaE*ab and A3, 17.58 +/- 3.33 DeltaE*ab), which were not different from each other. At 7 days: A1 was significantly different from A2; at 14 and 21 days: A2 and A3 were significantly better than A1; the Dunnett test revealed that the control group was different from A1, A2 and A3 at all periods (P < 0.05). CONCLUSION: Sodium perborate associated with both 10% and 35% carbamide peroxide was more effective than when associated with distilled water. PMID- 18422586 TI - Conventional and surgical endodontic retreatment of a maxillary lateral incisor with unusual anatomy. AB - AIM: To describe conventional and surgical endodontic treatment of a maxillary right lateral incisor with an unusual root dilaceration and to discuss the treatment outcome. SUMMARY: Root dilacerations refer to severe curvature or angulations that may occur anywhere along the length of a root. This anatomical deviation may present an unfavourable outcome if clinicians fail to recognize the difficulties. The following case report describes conventional and surgical endodontic treatment of a maxillary right lateral incisor with an unusual root dilaceration. *Root morphology may present a challenge for clinicians during root canal treatment. *Clinicians should be aware of the importance of surgical endodontics and be familiar with microsurgery principals and techniques. PMID- 18422587 TI - Pulp stones: a review. AB - Pulp stones are a frequent finding on bitewing and periapical radiographs but receive relatively little attention in textbooks. A review of the literature was therefore performed, initially using the PubMed database and beginning the search with 'pulp calcifications' and 'pulp stones'. Each term provided more than 400 references, many of which related to pulp calcification in general rather than pulp stones, and focussed largely on the problems these changes presented to clinicians. A manual search using references from this source was carried out. Contemporary textbooks in endodontology were also consulted, and an historic perspective gained from a number of older books and references. The factors involved in the development of the pulp stones are largely unknown. Further research may determine the reasons for their formation, but with current endodontic instruments and techniques this is unlikely to alter their relevance to clinicians. PMID- 18422588 TI - Safety and efficacy of home intravenous antimicrobial infusion therapy in older patients: a comparative study with younger patients. AB - BACKGROUND: Home intravenous antimicrobial infusion therapy has proved its safety and efficacy in a great number of infections. Despite this there are few published studies about this way of managing in the elderly patient. OBJECTIVE: To study the safety and efficacy of home intravenous antimicrobial infusion therapy in elderly patients. STUDY DESIGN: A prospective and comparative study of an elderly group of patients > or =70 years old vs. a cohort of younger adult patients as a control group. All patients were followed until 3 months after discharge. SETTING: Hospital at Home Programme (HHP) as part of the Internal Medicine Department at Valle de Hebron Hospital, Barcelona, Spain. PATIENTS: All patients admitted to HHP diagnosed of infections requiring intravenous antibiotic therapy between March 2006 and March 2007. RESULTS: We included 145 patients, 90 of whom were 70 years or older. Diabetes mellitus, heart failure and respiratory tract infection were more frequent in these elderly patients. In this group 14 (12%) developed some type of adverse event during treatment, phlebitis being the most common. The majority of those in the elderly patients group were discharged because of satisfactory clinical evolution and only 7 (7%) were re-admitted to hospital. Another 13 (14%) were re-admitted to hospital 3 months after discharge from HHP, mostly for chronic diseases. There were no significant differences between these results and those obtained from the control group. CONCLUSION: Home intravenous antimicrobial infusion therapy in elderly patients is safe and effective. PMID- 18422589 TI - Symptoms and signs of port-related infections in oncology patients related to the offending pathogens. AB - AIM: There is limited information about symptoms and signs of port-related infections linking to their offending pathogens. METHODS: Oncology patients proven to have port-related infections were retrospectively analysed. We divided them into two subgroups according to their symptoms and signs. Onset of fever and chills with or without hypotension following the port flush was classified as 'port flush form infection'. Presence of local inflammatory signs, including erythema, warmth, tenderness and pus formation and systemic infection signs, including fever, chills with or without hypotension was classified as 'local inflammatory form infection'. RESULTS: There were 29 episodes of port-related infection among 28 patients, with port flush form 22 episodes and local inflammatory form seven episodes. Of 22 episodes of port flush form infections, 20 (91%) were nosocomial glucose non-fermenting gram-negative bacilli, with Acinetobacter baumannii (11 episodes, 50%) and Enterobacter cloacae (four episodes, 18%) the most common. Polymicrobial infections occurred in four episodes (18%). Candida infection occurred in two episodes (9%). Of seven episodes of local inflammatory form infections, six (86%) were gram-positive cocci, with Staphylococcus aureus (five episodes, 71%) the most common. The time from port implantation to its infection was 272 +/- 255 days (30-993 days) for the port flush form infections and 82 +/- 87 days (22-265 days) for the local inflammatory form infections. This difference was not significant difference (p = 0.068). CONCLUSIONS: The differences between infection of patients with port flush form and local inflammatory form in incidence and offending microorganism suggest that the aetiology of infection were different. PMID- 18422590 TI - Head-to-head comparison of risedronate vs. teriparatide on bone turnover markers in women with postmenopausal osteoporosis: a randomised trial. AB - AIMS: We aimed to compare the effect of risedronate (RIS) and teriparatide (TPTD) (recombinant human parathyroid hormone 1-34) on bone turnover markers in women with postmenopausal osteoporosis. METHODS: Forty-four Caucasian women (age 65.1 +/- 1.6 years) with postmenopausal osteoporosis were randomly assigned to receive either RIS 35 mg once weekly (n = 22) or TPTD 20 microg once daily (n = 22) for 12 months. Serum N-terminal propeptide of type 1 collagen (P1NP), C-terminal telopeptide of type 1 collagen (CTx), total alkaline phosphatase (ALP) and intact parathyroid hormone (iPTH) were obtained from all women before, 3 and 6 months after treatment initiation. Lumbar spine bone mineral density (BMD) was measured by dual-energy X-ray absorptiometry before and 12 months after treatment initiation. RESULTS: P1NP, CTx and total ALP levels decreased in RIS group (p < 0.001) and increased in TPTD group (p < 0.001) throughout the treatment. iPTH increased significantly in RIS group (p < 0.05) and decreased in TPTD group (p < 0.001). Finally, lumbar spine BMD increased significantly in both RIS (p = 0.003) and TPTD groups (p < 0.001) without significant differences between them. CONCLUSIONS: Our data suggest that both serum P1NP and CTx are reliable markers of RIS and TPTD action in women with postmenopausal osteoporosis. In a similar way, serum total ALP can be used as an alternative marker for monitoring both RIS and TPTD action, while iPTH can be used only for TPTD-treated women. The increase in P1NP and CTx after 3 months of treatment with RIS or TPTD can predict the increase in BMD after 12 months of treatment. PMID- 18422591 TI - Clinical indicators which necessitate renal biopsy in type 2 diabetes mellitus patients with renal disease. AB - BACKGROUND: The diagnosis of diabetic nephropathy (DN) is always based on clinical grounds. However, the necessity for renal biopsy of type 2 diabetes mellitus (DM) patients with renal disease to establish the diagnosis remains unclear. METHODS: We retrospectively studied 50 type 2 diabetic patients performed with renal biopsy between December 2002 and December 2006. Based on renal pathology, patients were divided into group I: DN alone, group II: non diabetic renal disease (NDRD) superimposed on DN and group III: isolated NDRD. Factors like DM > 10 years, retinopathy, previous minimal proteinuria without sudden heavy proteinuria, no glomerular haematuria and non-small-sized kidney were collected to evaluate their sensitivity, specificity, positive predictive value and negative predictive value for prediction of DN or NDRD in type 2 diabetic patients. RESULTS: Group I consisted of 24 patients, group II 15 patients and group III 11 patients. Acute interstitial nephritis was the most prevalent second renal disease in our study. Sensitivity and specificity for group I was poor in five features except high sensitivity in no sudden heavy proteinuria (83.3%) and non-small-sized kidney (95.8%). Comparable retinopathy, sudden heavy proteinuria and haematuria (p > 0.05) was noted between the three groups. Significant biopsy indicators included higher serum albumin, lower urinary daily protein excretion and lower 24-h creatinine clearance (C(Cr)) rate (p < 0.05). CONCLUSION: Our study demonstrated that DM > 10 years and retinopathy did not exclude NDRD in type 2 DM patients, and need for renal biopsy. Higher serum albumin, lower urinary daily protein and 24-h C(Cr) were indicative for biopsy to exclude NDRD. PMID- 18422592 TI - Analgesic efficacy of intravenous paracetamol for outpatient fractional curettage: a randomised, controlled trial. AB - OBJECTIVE: To determine the efficacy of intravenous (i.v.) paracetamol for decreasing pain associated with fractional curettage. MATERIALS AND METHODS: This double-blind, randomised, placebo-controlled trial included 70 women who underwent fractional curettage. Patients were randomly assigned to receive i.v. infusion of either 1000 mg paracetamol (n = 36) or 0.9% normal saline as placebo (n = 34) prior to the procedure. The main outcome measure was the intensity of pain assessed by a 10-cm visual analogue scale. Pain scoring was performed at t(1) = prior to the procedure, t(2) = during the procedure and t(3) = 30 min after the procedure. Statistical analysis was performed using the Student's t test, chi-squared and Pearson's correlation analysis. RESULTS: The pain scores at t(2) and t(3) were significantly higher than the score at t(1) in both groups. There was no statistically significant difference between the pain scores at t(1), t(2) and t(3) among the two groups. Menopausal status had no effect on the pain scores and on the treatment outcome. The pain scores prior to the procedure were significantly higher in women with a history of endometrial curettage when compared with women who have no history. CONCLUSIONS: Fractional curettage is associated with mild pain. We were unable to detect a significant difference in pain scores in patients undergoing fractional curettage when comparing the use of i.v. paracetamol with placebo. History of prior endometrial curettage seems to be a predictor of the basal pain scores at admission although it does not affect the pain intensity related to the procedure. PMID- 18422593 TI - Investigation of early atherosclerotic changes in acromegalic patients. AB - BACKGROUND: Functional and morphological changes of endothelium were risk factors for mortality attributed to atherosclerosis. Studies investigating early atherosclerotic alterations and the effect of the treatment of acromegaly on these alterations gave conflicting results. OBJECTIVE: Surrogate markers of early atherosclerotic changes, i.e. brachial artery flow-mediated dilation (FMD) and carotid artery intima-media-thickness (IMT) in active and inactive acromegalic patients were compared with control subjects matched to patients for age, sex and cardiovascular risk factors to find out the direct effects of growth hormone (GH)/insulin-like growth factor-1 excess. METHODS: In 14 active acromegalics and their 14 matched controls, 14 inactive acromegalics and their 14 matched controls, carotid artery IMT and FMD of brachial artery were measured. Inactive acromegalics were in remission for at least 1 year. RESULTS: Active acromegalics had higher IMT than matched controls and inactive acromegalics (0.85 +/- 0.20 mm, 0.64 +/- 1.77 mm, 0.66 +/- 0.20 mm respectively; p < 0.005, p < 0.05) and IMT of inactive acromegalics was not different from their matched controls (0.61 +/- 0.12 mm). FMD was significantly lower in active acromegalics than in matched controls and inactive acromegalics (2.910 +/- 2.00 mm, 6.5 +/- 2.81 mm, 5.68 +/- 2.9 mm respectively; p < 0.005, p < 0.05). FMD of inactive acromegalics was not significantly different from their matched controls (7.96 +/- 3.12 mm). A significant inverse relationship was found between GH and FMD in active acromegalics (r = -0.659, p = 0.010). CONCLUSION: In active acromegalics, early atherosclerotic changes are not only attributed to the high prevalence of risk factors, but also to the abnormal GH secretion itself. PMID- 18422594 TI - Prevalence of primary Sjogren's syndrome in Turkey: a population-based epidemiological study. AB - OBJECTIVES: The aim of this study was to determine the prevalence of primary Sjogren's syndrome (pSS) in a general Turkish population according to the latest proposed American-European Consensus Group (AECG) criteria and European-1 (EU-1) criteria. METHODS: The study was conducted in two districts of Izmir and involved 2835 subjects 20 years of age and older. In the first stage, face-to-face interviews were performed at the registered households. In the second stage, subjects reporting symptoms of both dry eye and dry mouth were invited to the hospital for a full examination, which included Schirmer-1, sialometry and serologic tests. In the third stage, a minor salivary gland biopsy was performed as required. RESULTS: A total of 2887 subjects were contacted and a complete interview was obtained for 2835 (1551 female, 1284 male) subjects. A total of 159 subjects (126 female, 33 male) confirmed oral and ocular dryness, and 86 of these patients (54.1%) underwent a detailed clinical examination in the hospital. pSS was diagnosed in 10 patients (nine females) according to the EU-1 criteria, and in six patients (six females) according to the AECG criteria. We found a minimum crude prevalence of 0.21% [95% confidence interval (CI): 0.03-0.29] in the sample population and an age-sex adjusted prevalence of 0.16% (95% CI: 0.06-0.35), according to AECG criteria. According to EU-1 criteria, these prevalence rates were found to be 0.35% (95% CI: 0.10-0.45) and 0.28% (95% CI: 0.13-0.51) respectively. CONCLUSION: The pSS prevalence rates found in the Turkish population in this study were lower than the estimated prevalence rate in a general population. PMID- 18422595 TI - Human milk fortification and gastric emptying in the preterm neonate. AB - Fortification of expressed breast milk (EBM) is widely recommended for preterm feeding. Fortification of EBM results in increased caloric density and osmolarity, both of which may retard gastric emptying. As gastric emptying is a major determinant of feed tolerance, we investigated the effect of fortification (with Lactodex HMF) of EBM on gastric emptying in preterm neonates. The half gastric emptying time was measured using real time ultrasonography in 25 consecutive preterm neonates first on EBM alone, then on EBM + Lactodex HMF. Each baby served as its own control. The students t-test was used for statistical analysis. The mean gestation age was 34.48 +/- 0.77 weeks. The mean birth weight was 1.92 +/- 0.14 kg. The mean half gastric emptying time at an age of 5.4 +/- 0.86 days on EBM was 24.00 +/- 5.00 min and 24.40 +/- 5.06 min on EBM + human milk fortifier (HMF). The same at 2nd assessment (15.2 +/- 1.79 days), with EBM was 22.80 +/- 4.58 min vs. 23.60 +/- 4.89 min when given EBM + HMF. These differences were not statistically significant. Fortification of EBM with Lactodex HMF does not affect the gastric emptying in preterm neonates and therefore is unlikely to affect feed tolerance in them. PMID- 18422596 TI - Endovenous laser ablation for superficial venous insufficiency. AB - BACKGROUND: Endovenous laser ablation (EVLA) is a new minimally invasive alternative to conventional surgery for superficial venous insufficiency and varicose veins, where laser energy is used to ablate the incompetent veins. DISCUSSION: Endovenous laser ablation avoids the need for surgical incisions, and the complications of surgical exploration of the groin or popliteal fossa, and stripping. The procedure is commonly performed under local anaesthesia, with immediate mobilisation and rapid return to normal activity. Severe varicosity of tributaries may require adjunctive procedures such as microphlebectomy or sclerotherapy. CONCLUSION: Early outcomes and cosmesis are superior, and long term data is accumulating that recurrence of EVLA rates may be lower. PMID- 18422597 TI - Diode laser for paediatric airway procedures: a useful tool. AB - OBJECTIVE: A wide range of lasers have been used in the larynx. Diode laser is a portable and relatively inexpensive laser which is delivered via a glass fibre hand-held probe. The objective of this study was to report our experience with the use of diode laser in a variety of paediatric airway pathologies. METHODS: In this study, 90 diode laser laryngeal procedures were performed on 31 patients in the age range of 1 month to 16 years at the time of the operation. The follow up after the procedure has been in the range of 6 months to 3 years. RESULTS: As per our records 19/31 (61.3%) patients have been cured of their initial pathologies and were not under further review, 3/31 (9.6%) were having repeated laser treatments. The remaining 9/31 (29.0%) had to undergo further treatment. There was no laser-related intra-operative or postoperative complications. CONCLUSIONS: Diode laser is a good tool for several paediatric laryngeal pathologies. The ability to guide the laser light using the flexible glass fibre directly onto the area requiring vapourisation enables very precise treatment. PMID- 18422598 TI - Statins may reduce episodes of exacerbation and the requirement for intubation in patients with COPD: evidence from a retrospective cohort study. AB - INTRODUCTION: Statins have diverse anti-inflammatory effects in addition to their lipid-lowering ability. This study assesses the rate of chronic obstructive pulmonary disease (COPD) exacerbation and intubations in patients taking statins. METHODS: This is a retrospective cohort study of 185 patients with COPD exacerbation, with a 1-year follow-up. Outcomes examined were repeat hospitalisation and intubations for COPD exacerbation. Baseline characteristics for which the p-value was < or = 0.10 were considered as covariates for inclusion in a multivariate model. RESULTS: The statin group had fewer episodes of exacerbation and required intubation fewer times than the subjects not receiving statins (p < 0.0001 for both outcomes). Unadjusted odds ratios (OR) for no statin use vs. statin use were 9.54 (95% CI: 4.54-20.02) for exacerbation and 10.47 (CI: 4.56-24.01) for intubation. The OR, adjusted for the use of angiotensin converting enzyme inhibitors or angiotensin receptor blockers (ORa), were 2.35 (CI: 1.01-5.50) for non-statin users exhibiting an exacerbation and 10.36 (CI: 2.77-38.76) for this group requiring intubation, compared with statin users. Similarly, ORa for long-acting beta(2) agonists as a covariate were 3.01 (CI: 1.46-6.10) for exacerbation and 8.89 (CI: 3.67-21.32) for intubation. Time to outcome during the observation period was reduced by statins with the hazard ratio (HR) for exacerbation of 0.19 (CI: 0.06-0.14); HR for statins reducing intubation was 0.14 (95% CI: 0.10-0.30). CONCLUSIONS: These data suggest that the use of statins may be associated with lower incidence of both exacerbations and intubations in patients with COPD. PMID- 18422599 TI - Expression of inhibitors of apoptosis family protein in 7,12 dimethylbenz[a]anthracene-induced hamster buccal-pouch squamous-cell carcinogenesis is associated with mutant p53 accumulation and epigenetic changes. AB - Fifty outbred Syrian golden hamsters were equally divided into three experimental groups and two control groups. The pouches of the experimental groups were painted bilaterally with a 0.5% 7,12-dimethylbenz[a]anthracene (DMBA) solution thrice a week for 3, 7 and 14 weeks. One of the control groups was applied with mineral oil while another control group remained untreated throughout the experiment. Neither survivin nor cIAP2 could be detected in any of the control tissues, whereas survivin and cIAP2 were found to be significantly increased in 3 , 7- and 14-week DMBA-treated pouches compared with the control pouches. Expression of XIAP, cIAP1 and NAIP were noted for both the control and 3-, 7- and 14-week DMBA-treated pouches, but levels were found to be significantly elevated in the experimental groups compared with the control pouches. p53 was not detected in any control tissues, but was significantly increased in 3-, 7- and 14 week DMBA-treated pouches. Direct sequencing revealed a point mutation (C-->G) of p53 for pouch tissues treated with DMBA for 3 and 7 weeks, and there was a wide variation in the p53 sequence of the 14-week DMBA-treated pouch tissues, as compared with the control tissues. The control tissues had a survivin- and cIAP2 methylated allele, whereas the DMBA-treated tissues showed no evidence of survivin- and cIAP2-methylation. Neither the control nor DMBA-treated pouches showed evidence of XIAP-, cIAP1- or NAIP-methylation. Our results suggest that the expression of inhibitors of apoptosis family in DMBA-induced hamster buccal pouch squamous-cell carcinogenesis may be modulated by both genetic (mutant p53) and epigenetic mechanisms. PMID- 18422601 TI - Single dose intravenous thioacetamide administration as a model of acute liver damage in rats. AB - Thioacetamide (TAA) has been used extensively in the development of animal models of acute liver injury. Frequently, TAA is administered intraperitoneally to induce liver damage under anaesthesia. However, it is rarely administered by intravenous injection in conscious rats. The experiments in this study were designed to induce acute liver damage by single intravenous injection of TAA (0, 70 and 280 mg/kg) in unrestrained rats. Biochemical parameters and cytokines measured during the 60-h period following TAA administration, included white blood cells (WBC), haemoglobulin (Hb), platelet, aspartate transferase (GOT), alanine transferase (GPT), total bilirubin (TBIL), direct bilirubin (DBI), albumin, ammonia (NH3), r-glutamyl transpeptidase (r-GT), tumour necrosis factor alpha (TNF-alpha) and interleukin-6 (IL-6). Rats were sacrificed by decapitation 60 h after TAA administration and livers were removed immediately for pathology and immunohistochemical (IHC) examination. Another group of rats were sacrificed by decapitation 1, 6 and 24 h after TAA administration and livers were removed immediately for time course change of pathology and IHC examination. TAA significantly increased blood WBC, GOT, GPT, TBIL, DBIL, NH3, r-GT, TNF-alpha and IL-6 levels but decreased the blood Hb, platelet and albumin level. The levels of histopathological damage in the liver after intravenous TAA administration were also increased with a dose-dependent trend and more increased at 60 h after TAA administration. The levels of inducible nitric oxide synthase (iNOS) and nuclear factor-kappaB (NF-kappaB) detected by IHC in the liver after intravenous TAA administration were also increased with a dose-dependent trend and more increased at 1 h after TAA administration. Single intravenous TAA administration without anaesthesia is a restorable animal model which may be used to investigate acute liver damage. PMID- 18422602 TI - Schizosaccharomyces pombe Snf2SR, a novel SNF2 family protein, interacts with Ran GTPase and modulates both RanGEF and RanGAP activities. AB - Snf2SR, a suppressor of rna1(ts), which is a temperature-sensitive mutation in Schizosaccharomyces pombe RanGAP (GTPase activating protein), possesses both the SNF2 and the helicase domains conserved in the chromatin remodeling SNF2 ATPase/helicase protein family. We have now clarified a function of Snf2SR. Snf2SR indeed showed DNA-stimulated ATPase activity, proving that it is a member of the SNF2 ATPase/helicase family. Consistent with this role, Snf2SR was localized in the nucleus and cell fractionation analysis revealed that Snf2SR was tightly associated with the nuclear matrix. The disruption of snf2SR(+) was detrimental for a cell proliferation of S. pombe. Snf2SR that did not enhance RanGAP activity by itself, but abolished histone-H3-mediated RanGAP inhibition, as previously reported for the histone H3 methyltransferase, Clr4, another rna1(ts) suppressor. In contrast to Clr4, Snf2SR directly bound to the GDP-bound form of the S. pombe Ran homologue Spi1 and enhanced the nucleotide exchange activity of Pim1, the S. pombe RanGEF (guanine nucleotide exchange factor). Over expression of Spi1-G18V, a Ran GTPase mutant fixed in the GTP-bound form, was lethal to S. pombe Deltasnf2SR. Together, our results indicate that Snf2SR is involved in the Ran GTPase cycle in vivo. PMID- 18422600 TI - Elevated activity of the oxidative and non-oxidative pentose phosphate pathway in (pre)neoplastic lesions in rat liver. AB - (Pre)neoplastic lesions in livers of rats induced by diethylnitrosamine are characterized by elevated activity of the first irreversible enzyme of the oxidative branch of the pentose phosphate pathway (PPP), glucose-6-phosphate dehydrogenase (G6PD), for production of NADPH. In the present study, the activity of G6PD, and the other NADPH-producing enzymes, phosphogluconate dehydrogenase (PGD), isocitrate dehydrogenase (ICD) and malate dehydrogenase (MD) was investigated in (pre)neoplastic lesions by metabolic mapping. Transketolase (TKT), the reversible rate-limiting enzyme of the non-oxidative branch of the PPP, mainly responsible for ribose production, was studied as well. Activity of G6PD in (pre)neoplastic lesions was highest, whereas activity of PGD and ICD was only 10% and of MD 5% of G6PD activity, respectively. Glucose-6-phosphate dehydrogenase activity in (pre)neoplastic lesions was increased 25 times compared with extralesional parenchyma, which was also the highest activity increase of the four NADPH-producing dehydrogenases. Transketolase activity was 0.1% of G6PD activity in lesions and was increased 2.5-fold as compared with normal parenchyma. Transketolase activity was localized by electron microscopy exclusively at membranes of granular endoplasmic reticulum in rat hepatoma cells where G6PD activity is localized as well. It is concluded that NADPH in (pre)neoplastic lesions is mainly produced by G6PD, whereas elevated TKT activity in (pre)neoplastic lesions is responsible for ribose formation with concomitant energy supply by glycolysis. The similar localization of G6PD and TKT activity suggests the channelling of substrates at this site to optimize the efficiency of NADPH and ribose synthesis. PMID- 18422603 TI - Stau1 negatively regulates myogenic differentiation in C2C12 cells. AB - Sequential expression of myogenic regulatory factors (MRFs) such as MyoD and myogenin drives myogenic differentiation. Besides transcriptional activation of MRFs, this process is also coordinated by post-transcriptional regulation; MyoD and myogenin mRNAs are stabilized by RNA-binding protein HuR. Stau1 is known to regulate mRNA stability in a complex with Upf1, which is termed Stau1-mediated mRNA decay (SMD). We describe here that Stau1 is involved in the regulation of myogenesis. We found that knockdown of Stau1 promotes myogenesis including the expression of a muscle-specific marker protein, myoglobin, in C2C12 myoblasts. MyoD induces myogenin expression in response to induction of myogenesis, which is a key step to start myogenesis. The level of MyoD protein was not affected when Stau1 was depleted by siRNA, whereas the levels of myogenin mRNA and protein were increased in Stau1-knockdown cells. Interestingly, myogenin promoter activity was also increased in Stau1-knockdown cells in the absence of induction of myogenesis. Furthermore, Stau1-knockdown cells spontaneously progressed myogenesis including the expression of muscle-specific protein. Although Stau1 is involved in mRNA decay together with Upf1, Upf1-knockdown did not affect progression of myogenesis. Our results suggest that Stau1 negatively regulates myogenesis in C2C12 myoblasts through a mechanism that is different from SMD. PMID- 18422604 TI - Sequential activation of Rap1 and Rac1 small G proteins by PDGF locally at leading edges of NIH3T3 cells. AB - Moving cells form protrusions, such as filopodia and lamellipodia, and focal complexes at leading edges, which eventually enhance cell movement. The Rho family small G proteins, Rac1, Cdc42 and RhoA, are involved in the formation of these leading edge structures. We investigated the role of another small G protein Rap1 in the platelet-derived growth factor (PDGF)-induced formation of leading edge structures and cell movement. Upon stimulation of NIH3T3 cells by PDGF, leading edge structures were formed and Necl-5, integrin alpha(V)beta(3), and PDGF receptor were accumulated at leading edges. Rap1, upstream regulators of Rap1 such as Crk and C3G, and a downstream effector RalGDS, were accumulated at peripheral ruffles over lamellipodia. Over-expression of Rap1GAP, which inactivates Rap1, and knockdown of Rap1 inhibited the PDGF-induced formation of leading edge structures, accumulation of these molecules, and cell movement. In addition, Rap1 activation subsequently induced accumulation of Rac1, Vav2 and PAK at peripheral ruffles, which was inhibited by Rap1GAP and knockdown of Rap1. These results indicate that Rap1, activated by PDGF, is recruited to leading edges and that Rac1 is thereby activated locally at peripheral ruffles. This process is pivotal for the PDGF-induced formation of leading edge structures and cell movement. PMID- 18422605 TI - EMBRYO YELLOW gene, encoding a subunit of the conserved oligomeric Golgi complex, is required for appropriate cell expansion and meristem organization in Arabidopsis thaliana. AB - We identified an embryo yellow (eye) mutation in Arabidopsis that leads to the abnormal coloration and morphology of embryos. The eye mutant formed bushy plants, with aberrant organization of the shoot apical meristem (SAM) and unexpanded leaves with irregular phyllotaxy. The epidermal cells of the eye mutant were much smaller than that of the wild-type. Thus, EYE is required for expansion of cells and organs, and for formation of the organized SAM. Hydrophobic layers of epidermal cells were also disrupted, suggesting that EYE might be involved in the generation of the extra-cellular matrix. The mutated gene encoded a protein that is homologous to Cog7, a subunit of the conserved oligomeric Golgi (COG) complex, which is required for the normal morphology and function of the Golgi appratus. The eye mutation caused mislocalization of a Golgi protein. In addition, the size of the Golgi apparatus was also altered. Thus, EYE might be involved in transport or retention of Golgi-localized proteins and in maintenance of Golgi morphology. We propose that some Golgi-localized proteins, distributions of which are controlled by EYE, play important roles in expansion of cells and organs, and in formation of the properly organized SAM in plants. PMID- 18422606 TI - Rizatriptan efficacy in ICHD-II pure menstrual migraine and menstrually related migraine. AB - OBJECTIVE: To examine the efficacy of rizatriptan for the treatment of pure menstrual migraine (PMM). BACKGROUND: In 2004, the International Headache Society proposed new research criteria for menstrual migraine (International Classification of Headache Disorders [ICHD-II]). Two subtypes were defined: PMM, in which attacks occur exclusively with menstruation, and menstrually related migraine (MRM), in which attacks may also occur at other times of the cycle. METHODS: The 2 protocols (MM1 and MM2) were identical randomized, double-blind studies. Adult patients with ICHD-II menstrual migraine were assigned to either rizatriptan 10-mg tablet or placebo (2:1). Patients were to treat a single menstrual migraine attack of moderate or severe pain intensity. This prospectively planned substudy pooled data from patients with a diagnosis of PMM from both studies. The primary substudy endpoint was 2-hour pain relief. Efficacy data were summarized for patients with a diagnosis of MRM. RESULTS: Of 707 (MM1: 357, MM2: 350) patients treated in the study, 146 patients (MM1: 81, MM2: 65) had a diagnosis of PMM. The percentage of patients reporting 2-hour pain relief was significantly greater for rizatriptan than for placebo for both PMM (73% vs 50%, P = .006) and MRM subgroups (71% vs 52%, P < .001). Most other efficacy endpoints favored rizatriptan compared with placebo in patients with either PMM or MRM. CONCLUSION: Rizatriptan 10 mg was superior to placebo for the treatment of PMM, as measured by 2-hour pain relief. Rizatriptan was also effective for the treatment of MRM and for relief of migraine-associated symptoms for both headache subtypes. PMID- 18422607 TI - Predictors and correlates of edentulism in the healthy old people in Edinburgh (HOPE) study. AB - OBJECTIVES: To determine the extent to which correlates of edentulism are explained by an association between tooth loss and cognitive ability. METHODS: Participants in the Healthy Old People in Edinburgh (HOPE) study aged 70 or more at baseline were assessed and health, cognitive, socio-economic and socio environmental data collected on four consecutive occasions. It was noted whether the participant had any retained teeth and if not, the age when the last tooth was lost. Prior determinants of edentulism were investigated with binary logistic regression models. At the 9-year follow-up, associations with edentulism were examined using general linear models with edentulism as an independent factor. RESULTS: 201 participants were adequately tested, of whom 104 (51.7%) were edentulous. A logistic regression model that considered age, sex, education, social class, deprivation index of residence, objective distance from dentist, participant's estimate of distance from dentist and NART-estimated IQ (NARTIQ) found age (p = 0.032), occupational class (p = 0.019) and NARTIQ (p = 0.027) as significant predictors of edentulism. Cox's proportional hazards modelling found only NARTIQ (p = 0.050) to be correlated. Being edentulous was associated with poorer respiratory function but not hand grip strength (p = 0.23). Edentulous participants had lower self esteem scores (p = 0.020) and poorer dietary assessment scores (p = 0.028). Being edentulous was also associated with significantly lower mean scores on all cognitive testing, although these associations became non-significant after adjustment for NARTIQ and age. CONCLUSIONS: In healthy older people, edentulism is associated with relative impairment of cognitive ability, although this association is explained by the fact that lower original intelligence predisposes to edentulism and poorer performance on cognitive tests in old age. Once original intelligence is adjusted for, tooth loss is not related to cognitive ability. Tooth loss is, however, associated with poorer status across a wide range of health measures: physical health, nutrition, disability and self-esteem. Establishing the degree to which these health outcomes are causally related to edentulism could usefully be factored into cost-benefit analyses of programmes designed to prevent tooth loss. PMID- 18422608 TI - Experimental haemophilic arthropathy in a mouse model of a massive haemarthrosis: gross, radiological and histological changes. AB - Recurrent haemarthrosis results in chronic synovitis and destructive arthropathy. The long-term effect of a single haemorrhage is not known. To investigate the histopathological changes following a single, but major joint haemorrhage, an animal model of massive haemarthrosis without mechanical trauma was developed and is described in this manuscript. The knee joint capsule of mice deficient in coagulation factor VIII or IX and non-haemophilic wild type mice was punctured to induce a one time, but massive haemorrhage. The single joint puncture resulted in acute haemarthrosis in both types of haemophilic mice but not in wild type mice. Subsequent to injury, the changes in the knee joints were analysed using gross, histological and radiographic assessments and compared with the uninjured knee. In addition, a novel imaging modality, micro-computed tomography, was used to document the structural damage to the joint. Our results indicate that the long term changes classically observed in patients with advanced haemophilic arthropathy are evident following a single massive haemarthrosis. This model will allow a thorough investigation of the pathobiology of blood-induced joint disease and will be useful to test the efficacy of innovative therapeutic strategies to prevent haemophilic synovitis and arthropathy. PMID- 18422609 TI - Therapeutic shoulder arthroscopy in patients with clotting disorders. PMID- 18422610 TI - Cost-utility analysis of Canadian tailored prophylaxis, primary prophylaxis and on-demand therapy in young children with severe haemophilia A. AB - Primary prophylaxis is the emerging standard treatment for boys with severe haemophilia. Tailored (escalating-dose) prophylaxis (EscDose), beginning at a low frequency and escalating with repeated bleeding may prevent arthropathy at a lower cost than standard prophylaxis (SP). From a societal perspective, we compared the incremental cost per joint-haemorrhage that is avoided and quality adjusted-life-year (QALY) gained of SP and EscDose to on-demand (Demand) therapy in severe haemophilia A boys treated to age 6 using a decision analytic model. Costs included factor VIII (FVIII), professional visits and tests, central venous placement/complications, hospitalization, home programmes and parents' lost work days. Resource utilization was estimated by surveying 17 Canadian clinics. The natural history of bleeding and other probabilities were determined from a longitudinal chart review (n = 24) and published literature. EscDose costs an additional $3192 per joint-haemorrhage that was avoided compared with Demand whereas SP costs an additional $9046 per joint-haemorrhage that was avoided compared with EscDose. Clinic costs and lost wages were reduced by 60-80% for EscDose and SP compared with Demand. EscDose attained more QALYs than SP and Demand on account of less bleeding than Demand and lower need for ports than SP. The incremental cost per QALY for EscDose vs. Demand was $542 938. EscDose was less expensive with similar QALYs compared to SP. Sensitivity analysis was performed on all probability- and cost-estimates, and showed the model was sensitive to the cost of FVIII and the SP and target joint utilities. In conclusion, prophylaxis will substantially improve clinical outcomes and quality of life compared to Demand treatment, but with substantial cost. PMID- 18422612 TI - Guideline on the selection and use of therapeutic products to treat haemophilia and other hereditary bleeding disorders. A United Kingdom Haemophilia Center Doctors' Organisation (UKHCDO) guideline approved by the British Committee for Standards in Haematology. AB - Evidence-based guidelines are presented on the selection and use of therapeutic products to treat haemophilia and other hereditary bleeding disorders. They include details of therapeutic products available in the UK and they update and replace previous United Kingdom Haemophilia Centre Doctors' Organisation guidelines. PMID- 18422613 TI - A novel gene, ecl1(+), extends the chronological lifespan in fission yeast. AB - We have identified a novel gene from Schizosaccharomyces pombe that we have named ecl1(+) (extender of the chronological lifespan). When ecl1(+) is provided on a high-copy number plasmid, it extends the viability of both the Deltasty1 MAP kinase mutant and the wild-type cells after entry into the stationary phase. ecl1(+) encodes an 80-amino acid polypeptide that had not been annotated in the current database. The ecl1(+)-mRNA increases transiently when the growth phase is changed from the log phase to the stationary phase. The Ecl1 protein is localized in the nucleus. Calorie restriction extends the chronological lifespan of wild type and Deltaecl1 cells but not ecl1(+)-overproducing cells. The Deltapka1 mutant shows little, if any, additional extension of viability when Ecl1 is overproduced. The ste11(+) gene that is negatively controlled by Pka1 is up regulated when Ecl1 is overproduced. From these results we propose that the effect of Ecl1 overproduction may be mainly linked to and negatively affects the Pka1-dependent pathway. PMID- 18422614 TI - Candida ruelliae sp. nov., a novel yeast species isolated from flowers of Ruellia sp. (Acanthaceae). AB - Two novel yeast strains designated as 16Q1 and 16Q3 were isolated from flowers of the Ruellia species of the Acanthaceae family. The D1/D2 domain and ITS sequences of these two strains were identical. Sequence analysis of the D1/D2 domain of large-subunit rRNA gene indicated their relationship to species of the Candida haemulonii cluster. However, they differ from C. haemulonii by 14% nucleotide sequence divergence, from Candida pseudohaemulonii by 16.1% and from C. haemulonii type II by 16.5%. These strains also differ in 18 physiological tests from the type strain of C. haemulonii, and 12 and 16 tests, respectively, from C. pseudohaemulonii and C. haemulonii type II. They also differ from C. haemulonii and other related species by more than 13% sequence divergence in the internal transcribed spacer region. In the SSU rRNA gene sequences, strain 16Q1 differs by 1.7% nucleotide divergence from C. haemulonii. Sporulation was not observed in pure or mixed cultures on several media examined. All these data support the assignment of these strains to a novel species; we have named them as Candida ruelliae sp. nov., and designate strain 16Q1(T)=MTCC 7739(T)=CBS10815(T) as type strain of the novel species. PMID- 18422615 TI - Structure and genetics of Shigella O antigens. AB - This review covers the O antigens of the 46 serotypes of Shigella, but those of most Shigella flexneri are variants of one basic structure, leaving 34 Shigella distinct O antigens to review, together with their gene clusters. Several of the structures and gene clusters are reported for the first time and this is the first such group for which structures and DNA sequences have been determined for all O antigens. Shigella strains are in effect Escherichia coli with a specific mode of pathogenicity, and 18 of the 34 O antigens are also found in traditional E. coli. Three are very similar to E. coli O antigens and 13 are unique to Shigella strains. The O antigen of Shigella sonnei is quite atypical for E. coli and is thought to have transferred from Plesiomonas. The other 12 O antigens unique to Shigella strains have structures that are typical of E. coli, but there are considerably more anomalies in their gene clusters, probably reflecting recent modification of the structures. Having the complete set of structures and genes opens the way for experimental studies on the role of this diversity in pathogenicity. PMID- 18422616 TI - Diversity and occurrence of Burkholderia spp. in the natural environment. AB - Both in natural and in managed ecosystems, bacteria are common inhabitants of the phytosphere and the internal tissues of plants. Probably the most diverse and environmentally adaptable plant-associated bacteria belong to the genus Burkholderia. This genus is well-known for its human, animal and plant pathogenic members, including the Burkholderia cepacia complex. However, it also contains species and strains that are beneficial to plants and can be potentially exploited in biotechnological processes. Here we present an overview of plant associated Burkholderia spp. with special emphasis on beneficial plant Burkholderia interactions. A discussion of the potential for utilization of stable plant-Burkholderia spp. associations in the development of low-input cropping systems is also provided. PMID- 18422617 TI - From soil to gut: Bacillus cereus and its food poisoning toxins. AB - Bacillus cereus is widespread in nature and frequently isolated from soil and growing plants, but it is also well adapted for growth in the intestinal tract of insects and mammals. From these habitats it is easily spread to foods, where it may cause an emetic or a diarrhoeal type of food-associated illness that is becoming increasingly important in the industrialized world. The emetic disease is a food intoxication caused by cereulide, a small ring-formed dodecadepsipeptide. Similar to the virulence determinants that distinguish Bacillus thuringiensis and Bacillus anthracis from B. cereus, the genetic determinants of cereulide are plasmid-borne. The diarrhoeal syndrome of B. cereus is an infection caused by vegetative cells, ingested as viable cells or spores, thought to produce protein enterotoxins in the small intestine. Three pore forming cytotoxins have been associated with diarrhoeal disease: haemolysin BL (Hbl), nonhaemolytic enterotoxin (Nhe) and cytotoxin K. Hbl and Nhe are homologous three-component toxins, which appear to be related to the monooligomeric toxin cytolysin A found in Escherichia coli. This review will focus on the toxins associated with foodborne diseases frequently caused by B. cereus. The disease characteristics are described, and recent findings regarding the associated toxins are discussed, as well as the present knowledge on virulence regulation. PMID- 18422618 TI - Role of RNA polymerase sigma-factor (RpoS) in induction of glutamate-dependent acid-resistance of Escherichia albertii under anaerobic conditions. AB - Escherichia albertii is a potential enteric food-borne pathogen with poorly defined genetic and biochemical properties. Acid resistance is perceived to be an important property of enteric pathogens, enabling them to survive passage through stomach acidity so that they may colonize the mammalian gastrointestinal tract. We analyzed glutamate-dependent acid-resistance pathway (GDAR) in five E. albertii strains that have been identified so far. We observed that the strains were unable to induce GDAR under aerobic growth conditions. Mobilization of the rpoS gene restored aerobic induction of this acid-resistance pathway, indicating that all five strains may have a dysfunctional sigma-factor. On the other hand, under anaerobic growth conditions where GDAR is induced in an RpoS-independent manner (i.e. in Shigella spp. and Escherichia coli O157:H7 strains), only three out of five E. albertii strains successfully induced GDAR. The remainder of the two strains exhibited dependence on functional RpoS even under anaerobic conditions to express GDAR, a regulatory function previously considered to be redundant. The data indicate that certain E. albertii strains may have an alternate RpoS-dependent pathway for acid-resistance under anaerobic growth conditions. PMID- 18422619 TI - Stage-specific expression of Trypanosoma cruzi trans-sialidase involves highly conserved 3' untranslated regions. AB - trans-Sialidases (TSs) are virulence factors that allow some trypanosomatids to incorporate sialic acid from host molecules. Trypanosoma cruzi bears a complex gene family coding for TS members, which can be broadly divided into two groups: one translated in stages present in the mammalian host (trypomastigote TS, tTS) and one translated in the insect vector stages (epimastigote TS, eTS). The molecular basis underlying the expression of different, nonoverlapping sets of TS proteins in either host is poorly understood, particularly because of the lack of transcription initiation control in this organism. Here we show that 3' untranslated regions (3'UTRs) of tTS and eTS are highly conserved within each gene group but completely different between both groups. Importantly, tTS-3'UTR but not eTS-3'UTR promoted high expression of the green fluorescent protein reporter gene in the mammalian-dwelling stages. In epimastigotes, both 3'UTRs lead to a comparatively low expression of the reporter gene, although eTS-3'UTR was more efficient than tTS-3'UTR. These results stress the importance of posttranscriptional events, mainly driven by specific 3'UTRs, in gene expression regulation in T. cruzi. PMID- 18422620 TI - Different Mesorhizobium species associated with Caragana carry similar symbiotic genes and have common host ranges. AB - Fourteen strains representing 11 Caragana-nodulating Mesorhizobium genomic species were identified as representing Mesorhizobium amorphae, Mesorhizobium huakuii, Mesorhizobium septentrionale and groups related to Mesorhizobium plurifarium, Mesorhizobium temperatum, Mesorhizobium tianshanense and Mesorhizobium mediterraneum by sequencing of the 16S rRNA gene, 16S-23S internal transcribed spacer, partial housekeeping recA gene, and previously performed sodium dodecyl sulfate polyacrylamide gel electrophoresis of proteins and BOX-PCR fingerprinting. Despite their different taxonomic affiliation, highly similar symbiotic genes (>93% similarity for nodC and >91.8% similarity for nifH) were found among the Caragana strains and the three type strains for M. tianshanense, M. temperatum and M. septentrionale. Cross nodulation tests revealed that each of these 14 Caragana mesorhizobia and the three type strains mentioned above could effectively infect each of their original host plants, Caragana microphylla, Glycyrrhiza (host for M. tianshanense type strain) and Astragalus adsurgens (host for M. temperatum and M. septentrionale type strains). These results provide evidence that different Mesorhizobium species can nodulate with Caragana, and they have similar symbiotic genes (probably acquired by a phenomenon of lateral gene transfer) and common host ranges. PMID- 18422621 TI - Interaction of enterohemorrhagic Escherichia coli O157:H7 with mouse intestinal mucosa. AB - In this study, we used mouse ileal loops to investigate the interaction of enterohemorrhagic Escherichia coli (EHEC) O157:H7 with the mouse intestinal mucosa. With a dose of 10(9) and 3 h incubation, EHEC O157 was detected in the lumen and to a lesser extent associated with the epithelium. Typical attaching and effacing (A/E) lesions were seen, albeit infrequently. While the effector protein Tir was essential for A/E lesion formation, the bacterial type III secretion system adaptor protein TccP was dispensable. These results suggest that A/E lesions on mouse intestinal mucosa can be formed independently of robust actin polymerization. PMID- 18422622 TI - Microbial production of medium-chain-length 3-hydroxyalkanoic acids by recombinant Pseudomonas putida KT2442 harboring genes fadL, fadD and phaZ. AB - Monomers of microbial polyhydroxyalkanoates, mainly 3-hydroxyhexanoic acid (3HHx) and 3-hydroxyoctanoic acid (3HO), were produced by overexpressing polyhydroxyalkanoates depolymerase gene phaZ, together with putative long-chain fatty acid transport protein fadL of Pseudomonas putida KT2442 and acyl-CoA synthetase (fadD) of Escherichia coli MG1655 in P. putida KT2442. FadL(Pp), which is responsible for free fatty acid transportation from the extracellular environment to the cytoplasm, and FadD(Ec), which activates fatty acid to acyl CoA, jointly reinforce the fatty acid beta-oxidation pathway. Pseudomonas putida KT2442 (pYZPst01) harboring polyhydroxyalkanoates depolymerase gene phaZ of Pseudomonas stutzeri 1317 produced 1.37 g L(-1) extracellular 3HHx and 3HO in shake flask studies after 48 h in the presence of sodium octanoate as a sole carbon source, while P. putida KT2442 (pYZPst06) harboring phaZ(Pst), fadD(Ec) and fadL(Pp) achieved 2.32 g L(-1) extracellular 3HHx and 3HO monomer production under the same conditions. In a 48-h fed-batch fermentation process conducted in a 6-L fermentor with 3 L sodium octanoate mineral medium, 5.8 g L(-1) extracellular 3HHx and 3HO were obtained in the fermentation broth. This is the first time that medium-chain-length 3-hydroxyalkanoic acids (mcl-3HA) were produced using fadL(Pp) and fadD(Ec) genes combined with the polyhydroxyalkanoates depolymerase gene phaZ. PMID- 18422623 TI - Occurrence of qnr-positive clinical isolates in Klebsiella pneumoniae producing ESBL or AmpC-type beta-lactamase from five pediatric hospitals in China. AB - The plasmid-mediated quinolone resistance qnr genes in clinical isolates in adults have been described in different countries; however, the frequency of their occurrence has not been detected in pediatric patients. A total of 410 clinical isolates of Klebsiella pneumoniae, identified as producers of an extended-spectrum beta-lactamase (ESBL), or AmpC beta-lactamase, were collected from five children's hospitals in China during 2005-2006. The isolates were screened for the presence of the qnrA, qnrB, and qnrS genes, and then the harboring qnr gene isolates were detected for a bla gene coding for the TEM, SHV, CTX-M, and plasmid-mediated ampC gene by a PCR experiment. Ninety-two isolates (22.7%) were positive for the qnr gene, including 10 of qnrA (2.4%), 25 of qnrB (6.1%), and 62 of qnrS (15.1%). Eighty-one of the 92 (88.0%) qnr-positive isolates carried at least one bla gene for TEM, SHV, CTX-M, or DHA-1. The ciprofloxacin resistance increased 16-256-fold and oflaxacin resistance increased 2-32-fold in transconjugants, respectively. These results indicated that the plasmid-mediated qnr quinolone resistance gene was qnrS, followed by qnrB and qnrA. Most of the isolates also carried a bla gene coding ESBL or ampC gene coding DHA-1 among Klebsiella pneumoniae isolated from Chinese pediatric patients. PMID- 18422624 TI - High spatial resolution analysis of fungal cell biochemistry--bridging the analytical gap using synchrotron FTIR spectromicroscopy. AB - Fungi impact humans and the environment in many ways, for good and ill. Some fungi support the growth of terrestrial plants or are used in biotechnology, and yet others are established or emerging pathogens. In some cases, the same organism may play different roles depending on the context or the circumstance. A better understanding of the relationship between fungal biochemical composition as related to the fungal growth environment is essential if we are to support or control their activities. Synchrotron FTIR (sFTIR) spectromicroscopy of fungal hyphae is a major new tool for exploring cell composition at a high spatial resolution. Brilliant synchrotron light is essential for this analysis due to the small size of fungal hyphae. sFTIR biochemical characterization of subcellular variation in hyphal composition will allow detailed exploration of fungal responses to experimental treatments and to environmental factors. PMID- 18422625 TI - Animal models of mucosal Candida infection. AB - Rodent models of oral, vaginal and gastrointestinal Candida infection are described and discussed in terms of their scientific merits. The common feature of all experimental mucosal Candida infections is the need for some level of host immunocompromise or exogenous treatment to ensure quantitatively reproducible disease. A growing literature describes the contributions of such candidiasis models to our understanding of certain aspects of fungal virulence and host response to mucosal Candida albicans challenge. Evidence to date shows that T lymphocyte responses dominate host immune defences to oral and gastrointestinal challenge, while other, highly compartmentalized responses defend vaginal surfaces. By contrast the study of C. albicans virulence factors in mucosal infection models has only begun to unravel the complex of attributes required to define the difference between strongly and weakly muco-invasive strains. PMID- 18422626 TI - Differential gene expression and extracellular secretion of the collagenolytic enzymes by the pathogen Vibrio parahaemolyticus. AB - Vibrio parahaemolyticus, a causative agent of wound infections as well as food poisoning, harbors two collagenase genes: vppC and prtV. When cultivated at 26 degrees C in gelatin broth supplemented with 3.0% NaCl, significant collagenolytic activity was detected in the culture supernatant at the early stationary phase. Native polyacrylamide gel electrophoresis analysis revealed a 90-kDa protein, and N-terminal amino acid sequencing showed that this protein was VppC, generated through truncation of 72 N-terminal amino acid residues. Additionally, significant expression of only vppC was observed by reverse transcriptase PCR. By contrast, a vppC-negative mutant constructed through single crossover homologous recombination secreted a 50-kDa-collagenolytic enzyme; however, this enzyme was a serine protease that was reported previously. These results suggest that VppC is a primary extracellular collagenase produced by V. parahaemolyticus. PMID- 18422627 TI - Whole genome-based assessment of the taxonomic position of the arthropod pathogenic bacterium Rickettsiella grylli. AB - Rickettsiella grylli is an intracellular bacterial pathogen of aquatic and terrestrial arthropods. Previous determination of its 16S rRNA-encoding sequence has led to the taxonomic classification of the genus Rickettsiella in the class Gammaproteobacteria, order Legionellales, family Coxiellaceae, i.e. in close vicinity to vertebrate pathogenic bacteria of the genera Coxiella and Legionella. Here we use the additional information available from the recently published first whole genome sequence from this genus to evaluate critically the taxonomic classification of R. grylli beyond the 16S rRNA gene level. Using phylogenetic reconstruction, together with significance testing on a data basis defined by a core set of 211 previously identified families of protein-encoding genes, together with a reanalysis of 16S rRNA gene data, the present study firmly corroborates the assignment of this species to both the class Gammaproteobacteria and the order Legionellales. However, the results obtained from concatenated and single protein, single protein-encoding gene, and 16S rRNA gene data demonstrate a similar phylogenetic distance of R. grylli to both the Coxiellaceae and the Legionellaceae and are, therefore, inconsistent with its current family-level classification. Consequently, a respective reorganization of the order Legionellales is proposed. PMID- 18422628 TI - Biofilm formation and interactions of bacterial strains found in wastewater treatment systems. AB - Biofilm formation and adherence properties of 13 bacterial strains commonly found in wastewater treatment systems were studied in pure and mixed cultures using a crystal violet microtiter plate assay. Four different culture media were used, wastewater, acetate medium, glucose medium and diluted nutrient broth. The medium composition strongly affected biofilm formation. All strains were able to form pure culture biofilms within 24 h in at least one of the tested culture media and three strains were able to form biofilm in all four culture media, namely Acinetobacter calcoaceticus ATCC 23055, Comamonas denitrificans 123 and Pseudomonas aeruginosa MBL 0199. The adherence properties assessed were initial adherence, cell surface hydrophobicity, and production of amyloid fibers and extracellular polymeric substances. The growth of dual-strain biofilms showed that five organisms formed biofilm with all 13 strains while seven formed no or only weak biofilm when cocultured. In dual-strain cultures, strains with different properties were able to complement each other, giving synergistic effects. Strongest biofilm formation was observed when a mixture of all 13 bacteria were grown together. These results on attachment and biofilm formation can serve as a tool for the design of tailored systems for the degradation of municipal and industrial wastewater. PMID- 18422629 TI - Intragastric administration with recombinant Lactococcus lactis producing heme oxygenase-1 prevents lipopolysaccharide-induced endotoxemia in rats. AB - Gut injury is a pivotal initiating event in the dysfunctional inflammatory response that causes postinjury multiple organ failure. Heme oxygenase-1 (HO-1) is an important enzyme that provides cellular protection against oxidative stress in different in vitro and in vivo systems. In this study, we evaluated the protective effects of intragastrically administered live Lactococcus lactis secreting bioactive HO-1 to treat intestinal mucosal injury induced by lipopolysaccharide in rats. Intragastric administration with this recombinant L. lactis strain led to active delivery of HO-1 at the mucosa and significantly decreased morbidity and mortality of lipopolysaccharide -induced endotoxemia as confirmed by blinded macroscopic and microscopic inflammatory scores (Chiu's grade), myeloperoxidase activity, mortality, and tumor necrosis factor-alpha and IL-10 cytokine stimulation. This protective effect could be abolished by an HO-1 inhibitor, the zinc protoporphyrin-IX. Our results suggest that a food-grade bacterium genetically modified to deliver bioactive HO-1 in situ exerts a protective effect against intestinal mucosal injury in rats with endotoxemia via modulation of the immune system. This novel approach may be beneficial for the maintenance of the intestinal barrier and anti-inflammatory response of the lower intestine. PMID- 18422630 TI - Synergism among volatile organic compounds resulting in increased antibiosis in Oidium sp. AB - Oidium sp. has been recovered as an endophyte in Terminalia catappa (tropical chestnut) in Costa Rica. The volatile organic compounds (VOCs) of this organism uniquely and primarily consist of esters of propanoic acid, 2-methyl-, butanoic acid, 2-methyl-, and butanoic acid, 3-methyl-. The VOCs of Oidium sp. are slightly inhibitory to many plant pathogenic fungi. Previous work on the VOCs of Muscodor albus demonstrated that besides esters of small organic acids, a small organic acid and a naphthalene derivative were needed to obtain maximum antibiotic activity. Thus, the addition of exogenous volatile compounds such as isobutyric acid and naphthalene, 1,1'-oxybis caused a dramatic synergistic increase in the antibiotic activity of the VOCs of Oidium sp. against Pythium ultimum. In fact, at elevated concentrations, there was not only 100% inhibition of P. ultimum but killing as well. In addition, a coculture of Muscodor vitigenus (making only naphthalene) and Oidium sp. plus isobutyric acid produced an additive antibiosis effect against P. ultimum. The biological implications of multiple volatile compounds acting to bring about antibiosis in nature are discussed. PMID- 18422631 TI - Characterization of nucleoside triphosphate diphosphohydrolase activity in Trichomonas gallinae and the influence of penicillin and streptomycin in extracellular nucleotide hydrolysis. AB - Here we described an nucleoside triphosphate diphosphohydrolase (NTPDase) activity in living trophozoites of Trichomonas gallinae. The enzyme hydrolyzes a variety of purine and pyrimidine nucleoside di- and triphosphates in an optimum pH range of 6.0-8.0. This enzyme activity was activated by high concentrations of divalent cations, such as calcium and magnesium. Contaminant activities were ruled out because the enzyme was not inhibited by classical inhibitors of ATPases (ouabain, 5.0 mM sodium azide, oligomycin) and alkaline phosphatases (levamisole). A significant inhibition of ATP hydrolysis (38%) was observed in the presence of 20 mM sodium azide. Sodium orthovanadate inhibited ATP and ADP hydrolysis (24% and 78%), respectively. The apparent K(M) (Michaelis constant) values were 667.62+/-13 microM for ATP and 125+/-5.3 microM for ADP. V(max) (maximum velocity) values were 0.44+/-0.007 nmol Pi min(-1) per 10(6) trichomonads and 0.91+/-0.12 nmol Pi min(-1) per 10(6) trichomonads for ATP and ADP, respectively. Moreover, we showed a marked decrease in ATP, ADP and AMP hydrolysis when the parasites were grown in the presence of penicillin and streptomycin. The existence of an NTPDase activity in T. gallinae may be involved in pathogenicity, protecting the parasite from the cytolytic effects of the extracellular nucleotides. PMID- 18422632 TI - Effects of seven potential probiotic strains on specific immune responses in healthy adults: a double-blind, randomized, controlled trial. AB - This pilot study investigated the immunomodulatory properties of seven probiotic strains. Eighty-three healthy volunteers aged 18-62 years consumed 2 x 10(10) CFU of bacteria or a placebo (maltodextrin) over 3 weeks (D0-D21). Subjects received an oral cholera vaccine at D7 and at D14; blood and saliva samples were collected at D0, D21 and D28. Serum samples were analyzed for specific IgA, IgG and IgM, and saliva samples were analyzed for specific IgA only, by ELISA. Statistical analyses were based on Wilcoxon's signed-rank test (intragroup analyses) and exact median t-test (intergroup analyses). Salivary analysis showed no difference in specific IgA concentrations between groups. Serum analysis indicated an effect of some of the tested strains on specific humoral responses. Between D0 and D21, IgG increased in two probiotic groups, for example, Bifidobacterium lactis Bl-04 and Lactobacillus acidophilus La-14, compared with controls (P=0.01). Trends toward significant changes in immunoglobulin serum concentrations compared with controls (P<0.1) were found for six out of the seven probiotic strains. In conclusion, some strains of probiotics demonstrated a faster immune response measured with serum immunoglobulin indicators, especially IgG, although overall vaccination was not influenced. Specific strains of probiotics may thus act as adjuvants to the humoral immune response following oral vaccination. PMID- 18422633 TI - Comparison of deep-sea sediment microbial communities in the Eastern Mediterranean. AB - Bacterial and archaeal communities in sediments obtained from three geographically-distant mud volcanoes, a control site and a microbial mat in the Eastern Mediterranean deep-sea were characterized using direct 16S rRNA gene analyses. The data were thus in relation to the chemical characteristics of the (stratified) habitats to infer community structure-habitat relationships. The bacterial sequences in the different habitats were related to those of Actinobacteria, Bacilli, Chloroflexi, Alpha-, Beta-, Gamma-, Delta- and Epsilonproteobacteria and unclassified bacteria, including the JS1 group. The archaeal sequences found were affiliated with those of the Methanosarcinales, Thermoplasmales, Halobacteriales and Crenarchaea belonging to marine benthic group I and B, as well as MCG group archaea. In each sample, the communities were diverse and unique at the phylotype level. However, at higher taxonomic levels, similar groups were found in different sediments, and similar depth layers tended to contain similar communities. The sequences that dominated in all top layers (as well as in the mat) probably represented organisms involved in aerobic heterotrophy, sulfide-based chemoautotrophy and methanotrophy and/or methylotrophy. Sequences of organisms most likely involved in anaerobic methane oxidation, sulfate reduction and anaerobic heterotrophy were predominantly found in deeper layers. The data supported the notion of (1) uniqueness of each habitat at fine taxonomic levels, (2) stratification in depth and (3) conservation of function in the sediments. PMID- 18422634 TI - Thiazolidinedione derivatives in diabetes and cardiovascular disease: an update. AB - As the incidence and the public health impact of type 2 diabetes are constantly rising, treatment of hyperglycemia, prevention of diabetes-related complications are currently top medical priorities. Within the last decade several new classes of oral hypoglycemic agents were added to our armamentarium against diabetes. Among these new classes, the group of thiazolidinediones, which act through reduction of insulin resistance is perhaps the most widely used. For about 20 years, numerous background and clinical studies have evaluated the beneficial and adverse effects of these compounds. Current knowledge suggests that thiazolidinediones are as effective as metformin or sulfonylurea derivatives in improving glycemic control and exert several other beneficial metabolic and vascular effects, such as improvement in lipid profile, blood pressure lowering, redistribution of body fat away from the central compartment, microalbuminuria regression, reduction in subclinical vascular inflammation and others. On the other hand, currently used thiazolidinediones have well-established side effects, most important of which are fluid retention leading to weight gain and heart failure deterioration. Further, in the expectance of proper outcome studies to clarify the effects of these agents in cardiovascular morbidity and mortality, data from recent meta-analyses suggest that rosiglitazone may increase the risk for some cardiovascular outcomes. This article will discuss all the above issues attempting to provide an updated overview of this expanding field. PMID- 18422635 TI - Predicting natural mortality rates of plants and animals. AB - Understanding the factors that control the mortality rates of species in their natural environments is important for understanding the structure and dynamics of populations, communities and ecosystems. Here, we test a model of natural mortality that yields explicit, quantitative predictions based on the constraints of body size and temperature on individual metabolism. Extensive field data from plants, invertebrates, fish, birds and mammals indicate that much of the heterogeneity in rates of natural mortality can be predicted, despite the many extrinsic sources of mortality in natural systems. These results suggest that common 'rule(s)' govern mortality rates in ecological communities for organisms as diverse as plants and animals. PMID- 18422636 TI - Apparent competition can compromise the safety of highly specific biocontrol agents. AB - Despite current concern about the safety of biological control of weeds, assessing the indirect impacts of introduced agents is not common practice. Using 17 replicate food webs, we demonstrate that the use of a highly host-plant specific weed biocontrol agent, recently introduced into Australia, is associated with declines of local insect communities. The agent shares natural enemies (predators and parasitoids) with seed herbivore species from native plants, so apparent competition is the most likely cause for these losses. Both species richness and abundance in insect communities (seed herbivores and their parasitoids) were negatively correlated with the abundance of the biocontrol agent. Local losses of up to 11 species (dipteran seed herbivores and parasitoids) took place as the biocontrol agent abundance increased. Ineffective biocontrol agents that remain highly abundant in the community are most likely to have persistent, indirect negative effects. Our findings suggest that more investment is required in pre-release studies on the effectiveness of biocontrol agents, as well as in post-release studies assessing indirect impacts, to avoid or minimize the release of potentially damaging species. PMID- 18422637 TI - Disturbance-driven changes in the variability of ecological patterns and processes. AB - Understanding how disturbance shapes the dynamics of ecological systems is of fundamental importance in ecology. One emerging approach to revealing and appreciating disturbance effects involves examining disturbance-driven changes in the variability of ecological responses. Variability is rarely employed as a response variable to assess the influence of disturbance, but recent studies indicate that it can be an extremely sensitive metric, capturing differences obscured by averaging and conveying important ecological information about underlying causal processes. In this paper, we present a conceptual model to understand and predict the effects of disturbance on variability. The model estimates qualitative changes in variability by considering disturbance extent, frequency and intensity, as well as ecosystem recovery, and thereby captures not only the immediate effects of disturbance but also those that arise over time due to the biotic response to an event. We evaluate how well the model performs by comparing predictions with empirical results from studies examining a wide variety of disturbances and ecosystems, and discuss factors that may modify or even confound predictions. We include a concise guide to characterizing and detecting changes in variability, highlighting the most common and easily applied methods and conclude by describing several future directions for research. By considering variability as a response to disturbance, we gain another metric of fundamental system behaviour, an improved ability to identify organizing features of ecosystems and a better understanding of the predictability of disturbance driven change - all critical aspects of assessing ecosystem response to disturbance. PMID- 18422638 TI - History, chance and adaptation during biological invasion: separating stochastic phenotypic evolution from response to selection. AB - Introduced species often exhibit changes in genetic variation, population structure, selection regime and phenotypic traits as they colonize and expand into new ranges. For these reasons, species invasions are increasingly recognized as promising systems for studying adaptive evolution over contemporary time scales. However, changes in phenotypic traits during invasion occur under non equilibrium demographic conditions and may reflect the influences of prior evolutionary history and chance events, as well as selection. We briefly review the evidence for phenotypic evolution and the role of selection during invasion. While there is ample evidence for evolutionary change, it is less clear if selection is the primary mechanism. We then discuss the likelihood that stochastic events shift phenotypic distributions during invasion, and argue that hypotheses of adaptation should be tested against appropriate null models. We suggest two experimental frameworks for separating stochastic evolution from adaptation: statistically accounting for phenotypic variation among putative invasion sources identified by using phylogenetic or assignment methods and by comparing estimates of differentiation within and among ranges for both traits and neutral markers (Q(ST) vs. F(ST)). Designs that incorporate a null expectation can reveal the role of history and chance in the evolutionary process, and provide greater insights into evolution during species invasions. PMID- 18422639 TI - Beta-tropomyosin mutations alter tropomyosin isoform composition. AB - BACKGROUND AND PURPOSE: Tropomyosin (TM) is an actin-binding protein, which is localized head to tail along the length of the actin filament. There are three major TM isoforms in human striated muscle. Mutations in beta-tropomyosin (TPM2) have recently been identified as an important cause of neuromuscular disorders. MATERIALS AND METHODS: The expression of TM isoforms in patients carrying mutations in TPM2 was detected using a combination of SDS-PAGE, Western blotting, and a new method to measure the relative abundance of the various TM transcripts. RESULTS: The level of gamma-TM is reduced in patients with mutations in TPM2. Beta-tropomyosin was expressed at high levels in muscle specimens of the patients. DISCUSSION: Our study indicates that beta-TM gene mutations can alter the expression of other sarcomeric TM isoforms and that the perturbation of TM isoform levels may affect the dimer preference within the thin filaments, which may contribute to muscle weakness as a result of both functional and structural changes in muscle. PMID- 18422640 TI - Abundance and activity of Chloroflexi-type SAR202 bacterioplankton in the meso- and bathypelagic waters of the (sub)tropical Atlantic. AB - The contribution of Chloroflexi-type SAR202 cells to total picoplankton and bacterial abundance and uptake of D- and L-aspartic acids (Asp) was determined in the different meso- and bathypelagic water masses of the (sub)tropical Atlantic (from 35 degrees N to 5 degrees S). Fluorescence in situ hybridization (FISH) revealed that the overall abundance of SAR202 was < or = 1 x 10(3) cells ml(-1) in subsurface waters (100 m layer), increasing in the mesopelagic zone to 3 x 10(3) cells ml(-1) and remaining fairly constant down to 4000 m depth. Overall, the percentage of total picoplankton identified as SAR202 increased from < 1% in subsurface waters to 10-20% in the bathypelagic waters. On average, members of the SAR202 cluster accounted for about 30% of the Bacteria in the bathypelagic waters, whereas in the mesopelagic and subsurface waters, SAR202 cells contributed < 5% to total bacterial abundance. The ratio of D-Asp : L-Asp uptake by the bulk picoplankton community increased from the subsurface layer (D-Asp : L Asp uptake ratio approximately 0.03) to the deeper layers reaching a ratio of approximately 1 at 4000 m depth. Combining FISH with microautoradiography to determine the proportion of SAR202 cells taking up D-Asp versus L-Asp, we found that approximately 30% of the SAR202 cells were taking up L-Asp throughout the water column while D-Asp was essentially not taken up by SAR202. This D-Asp : L Asp uptake pattern of SAR202 cells is in contrast to that of the bulk bacterial and crenarchaeal community in the bathypelagic ocean, both sustaining a higher fraction of D-Asp-positive cells than L-Asp-positive cells. Thus, although the Chloroflexi-type SAR202 constitutes a major bathypelagic bacterial cluster, it does not contribute to the large fraction of d-Asp utilizing prokaryotic community in the meso- and bathypelagic waters of the North Atlantic, but rather utilizes preferentially L-amino acids. PMID- 18422641 TI - Molecular link of different stages of the trematode host of Neorickettsia risticii to Acanthatrium oregonense. AB - Neorickettsia risticii, the obligatory intracellular bacterium that causes Potomac horse fever, has been detected in various developmental stages of digenetic trematodes in the environment. Neorickettsia risticii-infected gravid trematodes were identified as Acanthatrium oregonense, based on morphologic keys. However, whether immature trematodes harbouring N. risticii are also A. oregonense was unknown. The objective of this study was to infer the life cycle of N. risticii-positive trematode hosts and transstadial transmission of the bacterium by molecularly characterizing the relationship among adult and immature stages of trematodes confirmed infected with N. risticii. Sequences of 18S ribosomal RNA genes up to 1922 bp in size were obtained from infected adult gravid trematodes, sporocysts and cercariae, and metacercariae. The sequences from the different immature stages of trematode are closely related to those of adult trematodes, some with 100% sequence identity; thus, they likely are life stages of A. oregonense. Comparisons with known 18S ribosomal RNA gene sequences of other digenetic trematodes indicated that all tested stages of the N. risticii positive trematodes belong to the family Lecithodendriidae, supporting the morphological identification. PMID- 18422642 TI - Isolation of novel bacteria, including a candidate division, from geothermal soils in New Zealand. AB - We examined bacterial diversity of three geothermal soils in the Taupo Volcanic Zone of New Zealand. Phylogenetic analysis of 16S rRNA genes recovered directly from soils indicated that the bacterial communities differed in composition and richness, and were dominated by previously uncultured species of the phyla Actinobacteria, Acidobacteria, Chloroflexi, Proteobacteria and candidate division OP10. Aerobic, thermophilic, organotrophic bacteria were isolated using cultivation protocols that involved extended incubation times, low-pH media and gellan as a replacement gelling agent to agar. Isolates represented previously uncultured species, genera, classes, and even a new phylum of bacteria. They included members of the commonly cultivated phyla Proteobacteria, Firmicutes, Thermus/Deinococcus, Actinobacteria and Bacteroidetes, as well as more-difficult to-cultivate groups. Isolates possessing < 85% 16S rRNA gene sequence identity to any cultivated species were obtained from the phyla Acidobacteria, Chloroflexi and the previously uncultured candidate division OP10. Several isolates were prevalent in 16S rRNA gene clone libraries constructed directly from the soils. A key factor facilitating isolation was the use of gellan-solidified plates, where the gellan itself served as an energy source for certain bacteria. The results indicate that geothermal soils are a rich potential source of novel bacteria, and that relatively simple cultivation techniques are practical for isolating bacteria from these habitats. PMID- 18422644 TI - Mutants of Saccharomyces cerevisiae deficient in acyl-CoA synthetases secrete fatty acids due to interrupted fatty acid recycling. AB - In the present study, acyl-CoA synthetase mutants of Saccharomyces cerevisiae were employed to investigate the impact of this activity on certain pools of fatty acids. We identified a genotype responsible for the secretion of free fatty acids into the culture medium. The combined deletion of Faa1p and Faa4p encoding two out of five acyl-CoA synthetases was necessary and sufficient to establish mutant cells that secreted fatty acids in a growth-phase dependent manner. The mutants accomplished fatty acid export during exponential growth-phase followed by fatty acid re-import into the cells during the stationary phase. The data presented suggest that the secretion is driven by an active component. The fatty acid re-import resulted in a severely altered ultrastructure of the mutant cells. Additional strains deficient of any cellular acyl-CoA synthetase activity revealed an almost identical phenotype, thereby proving transfer of fatty acids across the plasma membrane independent of their activation with CoA. Further experiments identified membrane lipids as the origin of the observed free fatty acids. Therefore, we propose the recycling of endogenous fatty acids generated in the course of lipid remodelling as a major task of both acyl-CoA synthetases Faa1p and Faa4p. PMID- 18422643 TI - Characterization of Aquifex aeolicus 4-diphosphocytidyl-2C-methyl-d-erythritol kinase - ligand recognition in a template for antimicrobial drug discovery. AB - 4-Diphosphocytidyl-2C-methyl-D-erythritol kinase (IspE) catalyses the ATP dependent conversion of 4-diphosphocytidyl-2C-methyl-D-erythritol (CDPME) to 4 diphosphocytidyl-2C-methyl-d-erythritol 2-phosphate with the release of ADP. This reaction occurs in the non-mevalonate pathway of isoprenoid precursor biosynthesis and because it is essential in important microbial pathogens and absent from mammals it represents a potential target for anti-infective drugs. We set out to characterize the biochemical properties, determinants of molecular recognition and reactivity of IspE and report the cloning and purification of recombinant Aquifex aeolicus IspE (AaIspE), kinetic data, metal ion, temperature and pH dependence, crystallization and structure determination of the enzyme in complex with CDP, CDPME and ADP. In addition, 4-fluoro-3,5-dihydroxy-4-methylpent 1-enylphosphonic acid (compound 1) was designed to mimic a fragment of the substrate, a synthetic route to 1 was elucidated and the complex structure determined. Surprisingly, this ligand occupies the binding site for the ATP alpha phosphate not the binding site for the methyl-D-erythritol moiety of CDPME. Gel filtration and analytical ultracentrifugation indicate that AaIspE is a monomer in solution. The enzyme displays the characteristic alpha/beta galacto-homoserine mevalonate-phosphomevalonate kinase fold, with the catalytic centre positioned in a deep cleft between the ATP- and CDPME-binding domains. Comparisons indicate a high degree of sequence conservation on the IspE active site across bacterial species, similarities in structure, specificity of substrate recognition and mechanism. The biochemical characterization, attainment of well-ordered and reproducible crystals and the models resulting from the analyses provide reagents and templates to support the structure-based design of broad-spectrum antimicrobial agents. PMID- 18422645 TI - A novel isoform of pantothenate synthetase in the Archaea. AB - The linear biosynthetic pathway leading from alpha-ketoisovalerate to pantothenate (vitamin B5) and on to CoA comprises eight steps in the Bacteria and Eukaryota. Genes for up to six steps of this pathway can be identified by sequence homology in individual archaeal genomes. However, there are no archaeal homologs to known isoforms of pantothenate synthetase (PS) or pantothenate kinase. Using comparative genomics, we previously identified two conserved archaeal protein families as the best candidates for the missing steps. Here we report the characterization of the predicted PS gene from Methanosarcina mazei, which encodes a hypothetical protein (MM2281) with no obvious homologs outside its own family. When expressed in Escherichia coli, MM2281 partially complemented an auxotrophic mutant without PS activity. Purified recombinant MM2281 showed no PS activity on its own, but the enzyme enabled substantial synthesis of [14C]4' phosphopantothenate from [14C]beta-alanine, pantoate and ATP when coupled with E. coli pantothenate kinase. ADP, but not AMP, was detected as a coproduct of the coupled reaction. MM2281 also transferred the 14C-label from [14C]beta-alanine to pantothenate in the presence of pantoate and ADP, presumably through isotope exchange. No exchange took place when pantoate was removed or ADP replaced with AMP. Our results indicate that MM2281 represents a novel type of PS that forms ADP and is strongly inhibited by its product pantothenate. These properties differ substantially from those of bacterial PS, and may explain why PS genes, in contrast to other pantothenate biosynthetic genes, were not exchanged horizontally between the Bacteria and Archaea. PMID- 18422646 TI - Gene expression silencing with 'specific' small interfering RNA goes beyond specificity - a study of key parameters to take into account in the onset of small interfering RNA off-target effects. AB - RNA-mediated gene silencing (RNA interference) is a powerful way to knock down gene expression and has revolutionized the fields of cellular and molecular biology. Indeed, the transfection of cultured cells with small interfering RNAs (siRNAs) is currently considered to be the best and easiest approach to loss-of function experiments. However, several recent studies underscore the off-target and potential cytotoxic effects of siRNAs, which can lead to the silencing of unintended mRNAs. In this study, we used a low-density microarray to assess gene expression modifications in response to five different siRNAs in various cell types and transfection conditions. We found major differences in off-target signature according to: (a) siRNA sequence; (b) cell type; (c) duration of transfection; and (d) post-transfection time before analysis. These results contribute to a better understanding of important parameters that could impact on siRNA side effects in knockdown experiments. PMID- 18422647 TI - Crystal structure of highly thermostable glycerol kinase from a hyperthermophilic archaeon in a dimeric form. AB - The crystal structure of glycerol kinase from the hyperthermophilic archaeon Thermococcus kodakaraensis (Tk-GK) in a dimeric form was determined at a resolution of 2.4 A. This is the first crystal structure of a hyperthermophilic glycerol kinase. The overall structure of the Tk-GK dimer is very similar to that of the Escherichia coli glycerol kinase (Ec-GK) dimer. However, two dimers of Ec GK can associate into a tetramer with a twofold axis, whereas those of Tk-GK cannot. This may be the reason why Tk-GK is not inhibited by fructose 1,6 bisphosphate, because the fructose 1,6-bisphosphate binding site is produced only when a tetrameric structure is formed. Differential scanning calorimetry analyses indicate that Tk-GK is a highly thermostable protein with a melting temperature (T(m)) of 105.4 degrees C for the major transition. This value is higher than that of Ec-GK by 34.1 degrees C. Comparison of the crystal structures of Tk-GK and Ec-GK indicate that there is a marked difference in the number of ion pairs in the alpha16 helix. Four ion pairs, termed IP1-IP4, are formed in this helix in the Tk-GK structure. To examine whether these ion pairs contribute to the stabilization of Tk-GK, four Tk-GK and four Ec-GK derivatives with reciprocal mutations at the IP1-IP4 sites were constructed. The determination of their stabilities indicates that the removal of each ion pair does not affect the stability of Tk-GK significantly, whereas the mutations designed to introduce one of these ion pairs stabilize or destabilize Ec-GK considerably. These results suggest that the ion pairs in the alpha16 helix contribute to the stabilization of Tk-GK in a cooperative manner. PMID- 18422648 TI - Structure and function of KH domains. AB - The hnRNP K homology (KH) domain was first identified in the protein human heterogeneous nuclear ribonucleoprotein K (hnRNP K) 14 years ago. Since then, KH domains have been identified as nucleic acid recognition motifs in proteins that perform a wide range of cellular functions. KH domains bind RNA or ssDNA, and are found in proteins associated with transcriptional and translational regulation, along with other cellular processes. Several diseases, e.g. fragile X mental retardation syndrome and paraneoplastic disease, are associated with the loss of function of a particular KH domain. Here we discuss the progress made towards understanding both general and specific features of the molecular recognition of nucleic acids by KH domains. The typical binding surface of KH domains is a cleft that is versatile but that can typically accommodate only four unpaired bases. Van der Waals forces and hydrophobic interactions and, to a lesser extent, electrostatic interactions, contribute to the nucleic acid binding affinity. 'Augmented' KH domains or multiple copies of KH domains within a protein are two strategies that are used to achieve greater affinity and specificity of nucleic acid binding. Isolated KH domains have been seen to crystallize as monomers, dimers and tetramers, but no published data support the formation of noncovalent higher-order oligomers by KH domains in solution. Much attention has been given in the literature to a conserved hydrophobic residue (typically Ile or Leu) that is present in most KH domains. The interest derives from the observation that an individual with this Ile mutated to Asn, in the KH2 domain of fragile X mental retardation protein, exhibits a particularly severe form of the syndrome. The structural effects of this mutation in the fragile X mental retardation protein KH2 domain have recently been reported. We discuss the use of analogous point mutations at this position in other KH domains to dissect both structure and function. PMID- 18422649 TI - Geraniol dehydrogenase, the key enzyme in biosynthesis of the alarm pheromone, from the astigmatid mite Carpoglyphus lactis (Acari: Carpoglyphidae). AB - Geraniol dehydrogenase (GeDH), which plays an important role in the biosynthesis of neral, an alarm pheromone, was purified from the astigmatid mite Carpoglyphus lactis. The enzyme was obtained in an apparently homogeneous and active form after 1879-fold purification through seven steps of chromatography. Car. lactis GeDH was determined to be a monomer in its active form with a relative molecular mass of 42 800, which is a unique subunit structure in comparison with already established alcohol dehydrogenases. Car. lactis GeDH oxidized geraniol into geranial in the presence of NAD+. NADP+ was ineffective as a cofactor, suggesting that Car. lactis GeDH is an NAD+-dependent alcohol dehydrogenase. The optimal pH and temperature for geraniol oxidation were determined to be pH 9.0 and 25 degrees C, respectively. The Km values for geraniol and NAD+ were 51.0 microm and 59.5 microm, respectively. Car. lactis GeDH was shown to selectively oxidize geraniol, whereas its geometrical isomer, nerol, was inert as a substrate. The high specificity for geraniol suggests that Car. lactis GeDH specializes in the alarm pheromone biosynthesis of Car. lactis. Car. lactis GeDH is composed of 378 amino acids. Structurally, Car. lactis GeDH showed homology with zinc-dependent alcohol dehydrogenases found in mammals and a mosquito (36.6-37.6% identical), and the enzyme was considered to be a member of the medium-chain dehydrogenase/reductase family, in view of the highly conserved sequences of zinc binding and NAD+-binding sites. Phylogenetic analyses indicate that Car. lactis GeDH could be categorized as a new class, different from other established alcohol dehydrogenases. PMID- 18422650 TI - Nuclear localization of human spermine oxidase isoforms - possible implications in drug response and disease etiology. AB - The recent discovery of the direct oxidation of spermine via spermine oxidase (SMO) as a mechanism through which specific antitumor polyamine analogues exert their cytotoxic effects has fueled interest in the study of the polyamine catabolic pathway. A major byproduct of spermine oxidation is H2O2, a source of toxic reactive oxygen species. Recent targeted small interfering RNA studies have confirmed that SMO-produced reactive oxygen species are directly responsible for oxidative stress capable of inducing apoptosis and potentially mutagenic DNA damage. In the present study, we describe a second catalytically active splice variant protein of the human spermine oxidase gene, designated SMO5, which exhibits substrate specificities and affinities comparable to those of the originally identified human spermine oxidase-1, SMO/PAOh1, and, as such, is an additional source of H2O2. Importantly, overexpression of either of these SMO isoforms in NCI-H157 human non-small cell lung carcinoma cells resulted in significant localization of SMO protein in the nucleus, as determined by confocal microscopy. Furthermore, cell lines overexpressing either SMO/PAOh1 or SMO5 demonstrated increased spermine oxidation in the nucleus, with accompanying alterations in individual nuclear polyamine concentrations. This increased oxidation of spermine in the nucleus therefore increases the production of highly reactive H2O2 in close proximity to DNA, as well as decreases nuclear spermine levels, thus altering the protective roles of spermine in free radical scavenging and DNA shielding, and resulting in an overall increased potential for oxidative DNA damage in these cells. The results of these studies therefore have considerable significance both with respect to targeting polyamine oxidation as an antineoplastic strategy, and in regard to the potential role of spermine oxidase in inflammation-induced carcinogenesis. PMID- 18422651 TI - Insights into substrate and product traffic in the Drosophila melanogaster acetylcholinesterase active site gorge by enlarging a back channel. AB - To test a product exit differing from the substrate entrance in the active site of acetylcholinesterase (EC 3.1.1.7), we enlarged a channel located at the bottom of the active site gorge in the Drosophila enzyme. Mutation of Trp83 to Ala or Glu widens the channel from 5 A to 9 A. The kinetics of substrate hydrolysis and the effect of ligands that close the main entrance suggest that the mutations facilitate both product exit and substrate entrance. Thus, in the wild-type, the channel is so narrow that the 'back door' is used by at most 5% of the traffic, with the majority of traffic passing through the main entrance. In mutants Trp83Ala and Trp83Glu, ligands that close the main entrance do not inhibit substrate hydrolysis because the traffic can pass via an alternative route, presumably the enlarged back channel. PMID- 18422652 TI - Molecular cloning and characterization of two soybean protein disulfide isomerases as molecular chaperones for seed storage proteins. AB - Protein disulfide isomerase family proteins play important roles in the folding of nascent polypeptides and the formation of disulfide bonds in the endoplasmic reticulum. In this study, we cloned two similar protein disulfide isomerase family genes from soybean leaf (Glycine max L. Merrill. cv Jack). The cDNAs encode proteins of 525 and 551 amino acids, named GmPDIL-1 and GmPDIL-2, respectively. Recombinant versions of GmPDIL-1 and GmPDIL-2 expressed in Escherichia coli exhibited oxidative refolding activity for denatured RNaseA. Genomic sequences of both GmPDIL-1 and GmPDIL-2 were cloned and sequenced. The comparison of soybean genomic sequences with those of Arabidopsis, rice and wheat showed impressive conservation of exon-intron structure across plant species. The promoter sequences of GmPDIL-1 apparently contain a cis-acting regulatory element functionally linked to unfolded protein response. GmPDIL-1, but not GmPDIL-2, expression was induced under endoplasmic reticulum-stress conditions. GmPDIL-1 and GmPDIL-2 promoters contain some predicted regulatory motifs for seed-specific expression. Both proteins were ubiquitously expressed in soybean tissues, including cotyledon, and localized to the endoplasmic reticulum. Data from coimmunoprecipitation experiments suggested that GmPDIL-1 and GmPDIL-2 associate with proglycinin, a precursor of the seed storage protein glycinin, and the alpha'-subunit of beta-conglycinin, a seed storage protein found in cotyledon cells under conditions that disrupt the folding of glycinin or beta-conglycinin, suggesting that GmPDIL-1 and GmPDIL-2 are involved in the proper folding or quality control of such storage proteins as molecular chaperones. PMID- 18422653 TI - Kinetic analysis of effector modulation of butyrylcholinesterase-catalysed hydrolysis of acetanilides and homologous esters. AB - The effects of tyramine, serotonin and benzalkonium on the esterase and aryl acylamidase activities of wild-type human butyrylcholinesterase and its peripheral anionic site mutant, D70G, were investigated. The kinetic study was carried out under steady-state conditions with neutral and positively charged aryl acylamides [o-nitrophenylacetanilide, o-nitrotrifluorophenylacetanilide and m-(acetamido) N,N,N-trimethylanilinium] and homologous esters (o-nitrophenyl acetate and acetylthiocholine). Tyramine was an activator of hydrolysis for neutral substrates and an inhibitor of hydrolysis for positively charged substrates. The affinity of D70G for tyramine was lower than that of the wild type enzyme. Tyramine activation of hydrolysis for neutral substrates by D70G was linear. Tyramine was found to be a pure competitive inhibitor of hydrolysis for positively charged substrates with both wild-type butyrylcholinesterase and D70G. Serotonin inhibited both esterase and aryl acylamidase activities for both positively charged and neutral substrates. Inhibition of wild-type butyrylcholinesterase was hyperbolic (i.e. partial) with neutral substrates and linear with positively charged substrates. Inhibition of D70G was linear with all substrates. A comparison of the effects of tyramine and serotonin on D70G versus the wild-type enzyme indicated that: (a) the peripheral anionic site is involved in the nonlinear activation and inhibition of the wild-type enzyme; and (b) in the presence of charged substrates, the ligand does not bind to the peripheral anionic site, so that ligand effects are linear, reflecting their sole interaction with the active site binding locus. Benzalkonium acted as an activator at low concentrations with neutral substrates. High concentrations of benzalkonium caused parabolic inhibition of the activity with neutral substrates for both wild-type butyrylcholinesterase and D70G, suggesting multiple binding sites. Benzalkonium caused linear, noncompetitive inhibition of the positively charged aryl acetanilide m-(acetamido) N,N,N-trimethylanilinium for D70G, and an unusual mixed-type inhibition/activation (alpha > beta > 1) for wild-type butyrylcholinesterase with this substrate. No fundamental difference was observed between the effects of ligands on the butyrylcholinesterase-catalysed hydrolysis of esters and amides. Thus, butyrylcholinesterase uses the same machinery, i.e. the catalytic triad S198/H448/E325, for the hydrolysis of both types of substrate. The differences in response to ligand binding depend on whether the substrates are neutral or positively charged, i.e. the differences depend on the function of the peripheral site in wild-type butyrylcholinesterase, or the absence of its function in the D70G mutant. The complex inhibition/activation effects of effectors, depending on the integrity of the peripheral anionic site, reflect the allosteric 'cross-talk' between the peripheral anionic site and the catalytic centre. PMID- 18422654 TI - Redox regulation of dimerization of the receptor protein-tyrosine phosphatases RPTPalpha, LAR, RPTPmu and CD45. AB - Whether dimerization is a general regulatory mechanism of receptor protein tyrosine phosphatases (RPTPs) is a subject of debate. Biochemical evidence demonstrates that RPTPalpha and cluster of differentiation (CD)45 dimerize. Their catalytic activity is regulated by dimerization and structural evidence from RPTPalpha supports dimerization-induced inhibition of catalytic activity. The crystal structures of CD45 and leukocyte common antigen related (LAR) indicate that dimerization would result in a steric clash. Here, we investigate dimerization of four RPTPs. We demonstrate that LAR and RPTPmu dimerized constitutively, which is likely to be due to their ectodomains. To investigate the role of the cytoplasmic domain in dimerization we generated RPTPalpha ectodomain (EDalpha)/RPTP chimeras and found that -- similarly to native RPTPalpha -- oxidation stabilized their dimerization. Limited tryptic proteolysis demonstrated that oxidation induced conformational changes in the cytoplasmic domains of these RPTPs, indicating that the cytoplasmic domains are not rigid structures, but rather that there is flexibility. Moreover, oxidation induced changes in the rotational coupling of dimers of full length EDalpha/RPTP chimeras in living cells, which were largely dependent on the catalytic cysteine in the membrane-distal protein-tyrosine phosphatase domain of RPTPalpha and LAR. Our results provide new evidence for redox regulation of dimerized RPTPs. PMID- 18422655 TI - Analysis of EphA4 receptor tyrosine kinase substrate specificity using peptide based arrays. AB - Eph receptor tyrosine kinases regulate many important biological processes. In the present study, we explored the substrate specificity of the EphA4 receptor tyrosine kinase using peptide arrays. We define a consensus substrate motif for EphA4 and go on to identify and test a number of potential EphA4 substrates and map their putative site(s) of phosphorylation. Cotransfection studies validate two of the predicted substrates: Nck2 and Dok1. Our findings identify several potential EphA4 substrates and demonstrate the general utility of using peptide arrays to rapidly identify and map protein kinase phosphorylation sites. PMID- 18422656 TI - An alternative transcript from the death-associated protein kinase 1 locus encoding a small protein selectively mediates membrane blebbing. AB - Death-associated protein kinase 1 (DAPK-1) is a multidomain protein kinase with diverse roles in autophagic, apoptotic and survival pathways. Bioinformatic screens were used to identify a small internal mRNA from the DAPK-1 locus (named s-DAPK-1). This encodes a 295 amino acid polypeptide encompassing part of the ankyrin-repeat domain, the P-loop motifs, part of the cytoskeletal binding domain of DAPK-1, and a unique C-terminal 'tail' extension not present in DAPK-1. Expression of s-DAPK-1 mRNA was detected in a panel of normal human tissues as well as primary colorectal cancers, indicating that its expression occurs in vivo. s-DAPK-1 gene transfection into cells produces two protein products: one with a denatured mass of 44 kDa, and a smaller product of 40 kDa. Double alanine mutation of the C-terminal tail extension of s-DAPK-1 (Gly296/Arg297) prevented production of the 40 kDa fragment, suggesting that the smaller product is generated by in vivo proteolytic processing. The s-DAPK-1 gene cannot substitute for full-length DAPK-1 in an mitogen-activated protein kinase kinase/extracellular signal-regulated kinase-dependent apoptotic transfection assay. However, the transfection of s-DAPK-1 was able to mimic full-length DAPK-1 in the induction of membrane blebbing. The 44 kDa protease-resistant mutant s DAPK-1G296A/R297A had very low activity in membrane blebbing, whereas the 40 kDa s-DAPK-1Deltatail protein exhibited the highest levels of membrane blebbing. Deletion of the tail extension of s-DAPK-1 increased its half-life, shifted the equilibrium of the protein from cytoskeletal to soluble cytosolic pools, and altered green fluorescent protein-tagged s-DAPK-1 protein localization as observed by confocal microscopy. These data highlight the existence of an alternative product of the DAPK-1 locus, and suggest that proteolytic removal of the C-terminal tail of s-DAPK-1 is required to stimulate maximally its membrane blebbing function. PMID- 18422657 TI - The role in the substrate specificity and catalysis of residues forming the substrate aglycone-binding site of a beta-glycosidase. AB - The relative contributions to the specificity and catalysis of aglycone, of residues E190, E194, K201 and M453 that form the aglycone-binding site of a beta glycosidase from Spodoptera frugiperda (EC 3.2.1.21), were investigated through site-directed mutagenesis and enzyme kinetic experiments. The results showed that E190 favors the binding of the initial portion of alkyl-type aglycones (up to the sixth methylene group) and also the first glucose unit of oligosaccharidic aglycones, whereas a balance between interactions with E194 and K201 determines the preference for glucose units versus alkyl moieties. E194 favors the binding of alkyl moieties, whereas K201 is more relevant for the binding of glucose units, in spite of its favorable interaction with alkyl moieties. The three residues E190, E194 and K201 reduce the affinity for phenyl moieties. In addition, M453 favors the binding of the second glucose unit of oligosaccharidic aglycones and also of the initial portion of alkyl-type aglycones. None of the residues investigated interacted with the terminal portion of alkyl-type aglycones. It was also demonstrated that E190, E194, K201 and M453 similarly contribute to stabilize ES(double dagger). Their interactions with aglycone are individually weaker than those formed by residues interacting with glycone, but their joint catalytic effects are similar. Finally, these interactions with aglycone do not influence glycone binding. PMID- 18422658 TI - Molecular determinants of ligand specificity in family 11 carbohydrate binding modules: an NMR, X-ray crystallography and computational chemistry approach. AB - The direct conversion of plant cell wall polysaccharides into soluble sugars is one of the most important reactions on earth, and is performed by certain microorganisms such as Clostridium thermocellum (Ct). These organisms produce extracellular multi-subunit complexes (i.e. cellulosomes) comprising a consortium of enzymes, which contain noncatalytic carbohydrate-binding modules (CBM) that increase the activity of the catalytic module. In the present study, we describe a combined approach by X-ray crystallography, NMR and computational chemistry that aimed to gain further insight into the binding mode of different carbohydrates (cellobiose, cellotetraose and cellohexaose) to the binding pocket of the family 11 CBM. The crystal structure of C. thermocellum CBM11 has been resolved to 1.98 A in the apo form. Since the structure with a bound substrate could not be obtained, computational studies with cellobiose, cellotetraose and cellohexaose were carried out to determine the molecular recognition of glucose polymers by CtCBM11. These studies revealed a specificity area at the CtCBM11 binding cleft, which is lined with several aspartate residues. In addition, a cluster of aromatic residues was found to be important for guiding and packing of the polysaccharide. The binding cleft of CtCBM11 interacts more strongly with the central glucose units of cellotetraose and cellohexaose, mainly through interactions with the sugar units at positions 2 and 6. This model of binding is supported by saturation transfer difference NMR experiments and linebroadening NMR studies. PMID- 18422659 TI - Biosynthesis of D-arabinose in mycobacteria - a novel bacterial pathway with implications for antimycobacterial therapy. AB - Decaprenyl-phospho-arabinose (beta-D-arabinofuranosyl-1-O-monophosphodecaprenol), the only known donor of d-arabinose in bacteria, and its precursor, decaprenyl phospho-ribose (beta-D-ribofuranosyl-1-O-monophosphodecaprenol), were first described in 1992. En route to D-arabinofuranose, the decaprenyl-phospho-ribose 2'-epimerase converts decaprenyl-phospho-ribose to decaprenyl-phospho-arabinose, which is a substrate for arabinosyltransferases in the synthesis of the cell-wall arabinogalactan and lipoarabinomannan polysaccharides of mycobacteria. The first step of the proposed decaprenyl-phospho-arabinose biosynthesis pathway in Mycobacterium tuberculosis and related actinobacteria is the formation of D ribose 5-phosphate from sedoheptulose 7-phosphate, catalysed by the Rv1449 transketolase, and/or the isomerization of d-ribulose 5-phosphate, catalysed by the Rv2465 d-ribose 5-phosphate isomerase. d-Ribose 5-phosphate is a substrate for the Rv1017 phosphoribosyl pyrophosphate synthetase which forms 5 phosphoribosyl 1-pyrophosphate (PRPP). The activated 5-phosphoribofuranosyl residue of PRPP is transferred by the Rv3806 5-phosphoribosyltransferase to decaprenyl phosphate, thus forming 5'-phosphoribosyl-monophospho-decaprenol. The dephosphorylation of 5'-phosphoribosyl-monophospho-decaprenol to decaprenyl phospho-ribose by the putative Rv3807 phospholipid phosphatase is the committed step of the pathway. A subsequent 2'-epimerization of decaprenyl-phospho-ribose by the heteromeric Rv3790/Rv3791 2'-epimerase leads to the formation of the decaprenyl-phospho-arabinose precursor for the synthesis of the cell-wall arabinans in Actinomycetales. The mycobacterial 2'-epimerase Rv3790 subunit is similar to the fungal D-arabinono-1,4-lactone oxidase, the last enzyme in the biosynthesis of D-erythroascorbic acid, thus pointing to an evolutionary link between the D-arabinofuranose- and L-ascorbic acid-related pathways. Decaprenyl phospho-arabinose has been a lead compound for the chemical synthesis of substrates for mycobacterial arabinosyltransferases and of new inhibitors and potential antituberculosis drugs. The peculiar (omega,mono-E,octa-Z) configuration of decaprenol has yielded insights into lipid biosynthesis, and has led to the identification of the novel Z-polyprenyl diphosphate synthases of mycobacteria. Mass spectrometric methods were developed for the analysis of anomeric linkages and of dolichol phosphate-related lipids. In the field of immunology, the renaissance in mycobacterial polyisoprenoid research has led to the identification of mimetic mannosyl-beta-1-phosphomycoketides of pathogenic mycobacteria as potent lipid antigens presented by CD1c proteins to human T cells. PMID- 18422660 TI - Dense smoke in the operating room: epivascular ultrasonography in a large right coronary artery aneurysm. PMID- 18422661 TI - Amorphous calcific tumor of the mitral annulus echocardiographically mimicking a vegetation. AB - We describe echocardiographic findings in an elderly patient with histologically proven amorphous tumor involving the posterior mitral annulus, mimicking a vegetation. PMID- 18422662 TI - Unusual left atrial appendage mass: atypical presentation of papillary fibroelastoma. AB - Papillary fibroelastomas are small, histologically benign neoplasms that are typically found on the valvular endocardium. We report a patient with a papillary fibroelastoma in an unusual location: the left atrial appendage. Although the mass was visualized both by computed tomography (CT) and transesophageal echocardiography, the diagnosis was histologically confirmed after surgical excision. PMID- 18422663 TI - Gender-dependent differences in echocardiographic characteristics of murine hearts. AB - Gender-dependent differences in echocardiographic characteristics of the heart have been thought to exist in human subjects, but comparisons have been difficult because of potential confounders such as differential prevalence of occult disease. Little data are available characterizing echocardiographic descriptors in normal experimental animals despite wide use of this modality. Accordingly, we characterized systolic and diastolic criteria of cardiac function echocardiographically in C57BL6 and BalbC age-matched mice to determine whether results in males and females differed. Left ventricular dimensions in absolute terms were lower, transmitral valve A-waves smaller and isovolumic relaxation time greater in females. When dimensional data were normalized for body weight, differences persisted but were directionally opposite. Thus, detection of abnormalities in cardiac function echocardiographically in various strains of mice requires establishing reference values that are gender as well as strain specific. Furthermore, the results suggest that gender-dependent differences are likely to be present in other species, including humans, and accordingly, that normal values should be established separately for males and females. PMID- 18422664 TI - Flying saucer located at the basal septum. AB - Left ventricular thrombus formation is a frequent complication in patients with ischemic heart disease and is associated with a high risk of systemic embolization. Generally, thrombi localize at the apical segment. However, thrombus localized at the basal septum has not been reported yet. In this case, we discuss a flying saucer shaped mass located at the basal septum, which was later diagnosed as thrombus after anticoagulant therapy. PMID- 18422665 TI - Relationship between left ventricular geometry and left ventricular systolic and diastolic functions in patients with chronic severe aortic regurgitation. AB - BACKGROUND: Chronic aortic regurgitation (AR) is a form of volume overload inducing left ventricle (LV) dilatation. Myocardial fibrosis, apoptosis, progressive LV dilatation, and eventually LV dysfunction are seen with the progression of disease. The aim of the study was to assess the relation between LV geometry and LV systolic and diastolic functions in patients with chronic severe AR. METHODS: The study population consisted of 88 patients with chronic severe AR and 42 healthy controls. The LV ejection fraction (LVEF) was calculated. Subjects were divided as Group I (controls, n = 42), Group II (LVEF > 50%, n = 47), and Group III (LVEF < 50%, n = 41). Transmitral early and late diastolic velocities and deceleration time were measured. The annular systolic (Sa) and diastolic (Ea and Aa) velocities were recorded. Diastolic function was classified as normal, impaired relaxation (IR), pseudonormalization (PN), and restrictive pattern (RP). RESULTS: The LVEF was similar in Group I and II, while significantly lower in Group III. Sa velocity was progressively decreasing, but LV long- and short-axis diameters were increasing from Group I to Group III. Forty-six, 31 and 11 patients had IR, PN, and RP, respectively. LV long-axis systolic and diastolic diameters were significantly increasing, while LVEF and Sa velocity were significantly decreasing from patients with IR to patients with RP. The LV long-axis diastolic diameter is independently associated with LV systolic and diastolic functions. CONCLUSIONS: The LV long-axis diastolic diameter is closely related with LV systolic and diastolic functions in patients with chronic severe AR. PMID- 18422666 TI - Evidence of robust coupling of atrioventricular mechanical function of the right side of the heart: insights from M-mode analysis of annular motion. AB - BACKGROUND: Extensive data exist regarding annular descent and ventricular function. We have already demonstrated significant differences in amplitude and timing of events between maximal mitral (MAPSE) and tricuspid (TAPSE) annular plane systolic excursion as well as described quantitative temporal differences in annular ascent (AA) between the right and left sides of the heart. However, whether any relationship exists between annular ascent and descent components remains uninvestigated. METHODS: Left ventricular ejection fraction (LVEF), right ventricular fractional area change (RVFAC), MAPSE, TAPSE, MV, and TV AA as well as pulsed tissue Doppler of the lateral MV and TV annuli were recorded from 53 patients. RESULTS: In this population (age 55 +/- 17 years) mean LVEF was 55 +/- 19%, mean RVFAC was 47 +/- 20%, mean MAPSE was 2.11 +/- 0.72 cm, mean TAPSE was 1.48 +/- 0.44 cm, mean MV AA was 0.52 +/- 0.17 cm, TV AA was 0.96 +/- 0.47, MV A wave 0.10 +/- 0.04 cm/s, and TV A-wave was 0.13 +/- 0.05 cm/s. A more robust correlation was seen between TV AA and RVFAC than between MV AA and LVEF and also between TV AA and pulsed TDI TV A-wave velocity than between MV AA and pulsed TDI MV A-wave. CONCLUSION: Our data reveal that mechanical systolic functions of the atria and the ventricles are more closely coupled on the right than on the left side of the heart. Whether this is a result of anatomic linking or chamber geometry will require further study. PMID- 18422667 TI - Quantitative assessment of myocardial acceleration in normal left ventricle with velocity vector imaging. AB - BACKGROUND: Application of two-dimensional myocardial acceleration map derived from tissue Doppler imaging is limited by inherent angle dependency and substantial reader variability in the visualization of the origin of ventricular activation site. In this study we investigated the characteristics of myocardial acceleration in normal left ventricular (LV) walls with velocity vector imaging (VVI). METHODS: VVI was applied to the parasternal short-axis two-dimensional echocardiographic images at basal, mid, and apical levels of the LV in 30 normal volunteers. Peak acceleration during early systole (ACC(s)) and time to ACC(s) (TACC(s)) were calculated for each segment of the standard 16-segment model. RESULTS: The time point of onset of active myocardial contraction corresponding to the QRS complex could not be determined in 409 (85.21%) of all 480 segments. No significant differences were found in TACC(s) among different LV levels and walls. In LV-free walls, there were no significant differences in ACC(s) among different LV levels and walls. CONCLUSIONS: The time point of onset of myocardial active contraction during early systole cannot be determined in most of normal myocardial segments. Also, there is homogeneity of the time to early systolic peak acceleration in the whole normal LV walls. Myocardial acceleration seems to have limited potential in the assessments of the site of initial electrical stimulation and the sequence of ventricular depolarization. PMID- 18422668 TI - Evaluation of right atrial size in patients with atrial arrhythmias: comparison of 2D versus real time 3D echocardiography. AB - AIM: This study compares different parameters of right atrial size evaluated by two-dimensional (2D) echocardiography with right atrial volume measured using three-dimensional echocardiography (3DE). METHODS AND RESULTS: One hundred sixty three consecutive patients with a history of atrial arrhythmias were studied by standard two-dimensional and by real time 3DE. Of these 142 (87%) recordings were of sufficient quality for interpretation of the right atrium by both imaging techniques. The following parameters of right atrial size were measured: apical four-chamber short-axis diameter (4CH short axis), apical four-chamber long axis diameter (4CH long axis), and apical four-chamber planimetry area. The 2D-derived right atrial volume was calculated by using the single plane area-length method (4CH area-length). The 2D parameters were then correlated with right atrial volume measured by real time 3DE. Linear regression analysis showed moderate correlation for four-chamber planimetry area (r = 0.72, P < 0.001) and 2D-derived volume calculation (r of 4CH single plane area-length RA volume = 0.70, P < 0.001). Diameters correlated clearly less well with 3DE volume (r of 4CH short axis = 0.61, 4CH long axis = 0.59, P < 0.001 respectively). CONCLUSION: Real time 3DE is highly feasible for right atrial volume determination. The results demonstrate that measurements of dimensions using 2D echocardiography may not accurately assess right atrial size. If 3DE is not available, apical 4CH planimetry area is a simple alternative that may be used for evaluating right atrial size in clinical practice. The 2D-derived right atrial volume by single plane area-length method was not better correlated with 3DE volume than four chamber planimetry area. PMID- 18422669 TI - Adult cardiac cystic lymphangiectasia of the right atrium. AB - Cardiac lympangiectasia is a rare pathological dilation of the lymphatic channels of the heart. Diagnosis is made by pathology; however, there remains no definitive diagnostic study. There are reports of cardiac cystic lymphangiectasias in children. A case of right atrial lymphangiectasia, initially identified by echocardiography, is presented here. This is the first report of such a mass. PMID- 18422670 TI - Isolated noncompaction of the left ventricular myocardium: a case report and literature review. AB - A previously healthy 30-year-old African American male presented to our hospital with diabetic ketoacidosis as his first manifestation of diabetes. A transthoracic echocardiogram (TTE) was performed with contrast, which showed isolated noncompaction of the left ventricular myocardium (INVM). The case presented here is followed by an extensive review of the available literature. PMID- 18422671 TI - Athletes' heart and echocardiography: athletes' heart. AB - Sudden death of competitive athletes is rare. However, they continue to have an impact on both the lay and medical communities. These deaths challenge the perception that trained athletes represent the healthiest segment of modern society. There is an increasing frequency of such reported deaths worldwide and the visibility of this issue is underlined by the high-profile nature of each case. Differential diagnosis between pathological and the physiologic (nonpathological) responses to high levels of physical training has become clinically more important. The purpose of this review is to highlight the main echocardiograph characteristics related to different types of training/sports participation and to highlight already recognized and newer concepts in their clinical assessment. PMID- 18422672 TI - Effects of radiofrequency catheter ablation on myocardial performance index and plasma NT-Pro-BNP levels in patients with Wolff-Parkinson-White syndrome. AB - BACKGROUND: The myocardial performance index (Tei index) is an echocardiographic index of combined systolic and diastolic functions. Brain natriuretic peptide (BNP) and its biologically inactive fragment N-terminal pro-BNP (NT-pro-BNP) are secreted by the heart in response to myocardial stretch. In this study, we investigated Tei index and NT-pro-BNP levels in patients with Wolff-Parkinson White (WPW) syndrome before and after radiofrequency catheter ablation therapy (RFCA). METHODS: Thirty patients (19 males, 11 females, aged 35.5 +/- 14.4 years) with WPW syndrome were enrolled in this study. Echocardiographic examination was performed before and 24 hours after RFCA. Tei index was calculated using Doppler echocardiography. Blood samples were taken before and 24 hours after RFCA to detect levels of NT-pro-BNP. RESULTS: Although isovolumic contraction time (IVCT) and isovolumic relaxation time (IVRT) did not change, aortic ejection time (ET) was decreased after RFCA (276 +/- 22 ms vs 254 +/- 30 ms, P < 0.01). So Tei index was significantly higher in postablation period (0.36 +/- 0.11 vs 0.42 +/- 0.21, P < 0.05). NT-pro-BNP levels did not change significantly after RFCA. CONCLUSIONS: We demonstrated that restoration of normal atrioventricular conduction by RFCA, leads to increase in Tei index but does not effect plasma NT pro-BNP levels. PMID- 18422673 TI - Idiopathic premature closure of the ductus arteriosus: an indication for early delivery. AB - Idiopathic intrauterine closure of the ductus arteriosus, distinct from intrauterine constriction associated with maternal NSAID ingestion or other secondary causes, is an uncommon event that often results in significant morbidity and mortality. We report on a 32-week fetus with meconium pseudocyst that presented with idiopathic intrauterine constriction of the ductus arteriosus, and review the literature. We suggest that early diagnosis and judicious obstetrical management, in conjunction with pediatric cardiology consultation based on serial fetal echocardiography, can lead to timely delivery and reversal of the pathophysiology providing the potential for a favorable outcome. PMID- 18422674 TI - Echocardiographic diastolic dysfunction and magnetic resonance infarct size in healed myocardial infarction treated with primary angioplasty. AB - BACKGROUND: After acute myocardial infarction (MI) the severity of diastolic dysfunction by echocardiography represents an independent prognostic marker. However, the mechanisms whereby diastolic dysfunction portends an increased risk after MI are not fully understood. We investigated the relationship between echocardiographic diastolic dysfunction severity and infarct size quantitatively measured by contrast-enhanced magnetic resonance (ce-MR). METHODS: Cross sectional prospective study. We quantified "healed" infarct size by ce-MR measuring the percentage of delayed enhancement with respect to left ventricular mass and diastolic function by Doppler echocardiography. Both exams were scheduled at least 1 month after a first acute ST segment elevation MI (STEMI) successfully treated with primary angioplasty and stenting. To increase the specificity, individual echocardiographic parameters were integrated to grade global diastolic function in 4 grades: normal diastolic function, impaired relaxation with normal, or near-normal filling pressures; impaired relaxation with moderate elevation of filling pressures, and impaired relaxation with marked elevation of filling pressures, "restrictive filling." RESULTS: We prospectively enrolled 52 patients (mean age 62 +/- 13 years, 77% men). ce-MR and echocardiography were performed 48 +/- 15 days after the MI. There was a significant but modest correlation between diastolic function grade and infarct size (r = 0.423, P = 0.002), which was independent of global and regional systolic function and persisted after further adjustment for age, sex, body surface area, left ventricular mass, end-diastolic volumes, and sphericity index (all P < 0.05). Among single echocardiographic variables, infarct size correlated best with tissue Doppler velocities Em (r =-0.307, P = 0.03), Am (r =-0.39, P = 0.005), and flow propagation velocity (r =-0.34, P = 0.015). CONCLUSIONS: In healed STEMI successfully treated with primary angioplasty and stenting, diastolic function grade was independently albeit weakly correlated with infarct size. Therefore, the increased risk of diastolic dysfunction after MI is not fully explained by infarct size. PMID- 18422675 TI - Adverse effects of dipeptidyl peptidases 8 and 9 inhibition in rodents revisited. AB - AIM: To evaluate the association between inhibition of dipeptidyl peptidase (DPP) 8 and/or DPP-9 organ toxicities and mortality in rodents. RESEARCH DESIGN AND METHODS: The relative selectivity of the DPP-4 inhibitor, vildagliptin, was determined by comparing its K(I) (concentration of compound yielding 50% inhibition of the enzyme) values for inhibition of recombinant human DPP-4, DPP-8 and DPP-9 assessed in vitro. In experiments performed in vivo, vildagliptin was administered by gavage for 13 weeks, at doses up to 1500 mg/kg/day in CD-1 mice and at doses up to 900 mg/kg/day in Wistar rats. Plasma concentrations of vildagliptin were assessed at week 12, and toxicities previously ascribed to inhibition of DPP-8 and/or DPP-9 were assessed at week 13. RESULTS: The K(I) values for vildagliptin-induced inhibition of DPP-4, DPP-8 and DPP-9 were 3, 810 and 95 nM respectively. The mean plasma concentration 24 h after dose after 12 week daily dosing with 1500 mg/kg/day in mice was 2279 nM. The mean plasma drug level 24 h after dose after 12-week daily dosing with 900 mg/kg/day in rats was 5729 nM. These high doses maintained plasma drug levels well above the K(I) values for DPP-8 and DPP-9 throughout a 24-h period. At these high doses, the toxicities of a selective DPP-8/DPP-9 inhibitor that were reported previously (100% mortality in mice, alopecia, thrombocytopenia, reticulocytopenia, enlarged lymph nodes, splenomegaly and 20% mortality in rats) were not observed. CONCLUSIONS: Inhibition of DPP-8 and DPP-9 per se does not lead to organ toxicities and mortality in rodents. Thus, a mechanism other than DPP-8/DPP-9 inhibition likely underlies the toxicity previously reported to be associated with a selective DPP-8/DPP-9 inhibitor. PMID- 18422676 TI - Respiratory patterns and strategies during feeding in preterm infants. AB - Because patterns of integration of respiration into rhythmic suck-swallow efforts are highly variable, we examined the vagaries of respiratory efforts as they evolve from the first tentative attempts at integration through more complex rhythmic interactions, with a focus on several strategies in which breathing and suck-swallow are coordinated. Thirty-four preterm infants (18 males, 16 females) of 26 to 33 weeks gestational age, 32 to 40 weeks postmenstrual age (PMA), and 2 to 12 weeks postnatal age were studied weekly from initiation of bottle feeding (using breast milk or preterm formula, both fed from a bottle) until discharge, with simultaneous digital recordings of pharyngeal pressure, nasal thermistor airflow, and thoraco-abdominal strain-gauge readings. Exceptional patterns of feeding-adapted variations of respiration were noted, including breathing during swallow, alternating blocks of suck-swallow and respiration efforts, narial airflow without thoracic movement, modulation of respiratory phase relationship against swallow rhythm, and paired rhythms with swallow:breath ratios of more than 1:1. Some of these strategies were developmentally regulated. Alternating blocks of suck-swallow and respiratory efforts were only seen in the earliest (PMA 32-33 wks) studies. In contrast, coordination and phase relationships of suck-swallow and breathing stabilized over time, as did the percentage of synchronized narial and thoracic respiratory efforts, which increased significantly after 36 weeks PMA compared with synchronization at 32 to 33.9 and 34 to 35.9 weeks PMA (p<0.05). There was also a significant positive correlation between percentage synchronization and PMA (r=0.58; p<0.001). The strategies and patterns noted here further clarify the developmentally regulated coordination of suck, swallow, and respiration into mature infant feeding, and may be predictive of those infants with short- and long-term feeding or developmental difficulties. PMID- 18422678 TI - Chronic post-traumatic headache after head injury in children and adolescents. AB - This was a prospective, observational study of children aged 3 to 15 years admitted to hospital with head injury (HI). Demographic data and information on the nature of the HI, and history of premorbid headache were collected. A structured telephone questionnaire was used to interview parents and children 2 months after injury and at 4-monthly intervals for up to 3 years, if headache was reported. One hundred and ninety children were admitted with HI. Data were available on 117 children (81 males, 36 females; mean age 8y 5mo [SD 3y 1mo]). HI was minor in 93 patients and significant in the rest. Minor HI was defined as a closed injury, no loss of consciousness, and a Glasgow Coma Score (GCS) of 13 to 15. Significant HI was associated with loss of consciousness for >30 minutes, GCS of <13, and post-traumatic amnesia for >48 hours. Eight children (five males, three females; mean age 10y 7mo [SD 2y]) reported chronic post-traumatic headache (CPTH). Five children had episodic tension-type headache and three had migraine with or without aura. Headache resolved over 3 to 27 months in all except one child who was lost to follow-up. Premorbid headache in three children transformed in frequency and type following HI. These patients were excluded from the study. CPTH is common after minor and significant HI. It has the clinical features of tension-type headache and migraine and has a good prognosis. PMID- 18422677 TI - Growth failure in children with intractable epilepsy is not due to increased resting energy expenditure. AB - The aim of this study was to evaluate the resting energy expenditure (REE) of children with intractable epilepsy (IE) compared with healthy children, and to determine factors that contribute to the pattern of REE. REE, growth status, and body composition were assessed in 25 prepubertal children with IE (15 males, 10 females; mean age 5y 5mo [SD 2y 2mo] range 2-9y) with and without cerebral palsy (CP) and compared with those in 75 healthy children of similar age, sex, and fat free mass (FFM; 43 males, 32 females; mean age 6y 4mo [SD 1y 8mo], range 2-9y). Of the 25 children with IE, 12 had generalized and 13 partial seizures; 10 children had CP (four hemiplegia, one diplegia, and five tetraplegia); 18 were ambulators. REE (kcal/d), determined by indirect calorimetry, was expressed as a percentage of that predicted using Schofield equations. Energy intake from 3-day weighed food records was assessed for children with IE only and expressed as a percentage of estimated energy requirement. Compared with healthy children, children with IE had significantly lower percentage (Student's t-test, p<0.05) of predicted REE (111 [SD 13] vs 104 [SD 4]), weight z-score, body mass index z score, and FFM. Using multiple regression, REE adjusted for FFM, fat mass, and sex were significantly lower in children with IE and CP (-110 kcal/d, 95% confidence interval -199 to -21, p=0.016). In children with IE, energy intake was also a statistically significant predictor of REE. CP largely explained the suboptimal growth status and lower REE of children with IE compared with healthy children. PMID- 18422679 TI - Aicardi-Goutieres syndrome: an important Mendelian mimic of congenital infection. AB - Aicardi-Goutieres syndrome (AGS) is a rare, genetically determined encephalopathy whose importance from a clinical viewpoint is magnified because of the risk of misdiagnosis as the sequelae of congenital infection. Recent molecular advances have shown that AGS can be caused by mutations in any one of at least five genes (four of which have so far been identified), most commonly on a recessive basis but occasionally as a dominant trait. Additionally, a recent genotype-phenotype correlation has shown that two clinical presentations can be delineated; an early onset neonatal form highly reminiscent of congenital infection seen particularly with TREX1 mutations, and a later-onset presentation, sometimes occurring after several months of normal development and occasionally associated with remarkably preserved neurological function, most frequently due to RNASEH2B mutations. Evidence is emerging to show that the nucleases defective in AGS are involved in removing endogenous nucleic acid species produced during normal cellular processing, and that a failure of this removal results in inappropriate activation of the innate immune system. This hypothesis explains the phenotypic overlap of AGS with congenital infection and some aspects of systemic lupus erythematosus, where a similar interferon alpha-mediated innate immune response is triggered by viral and host nucleic acids respectively. PMID- 18422680 TI - The King's Outcome Scale for Childhood Head Injury and injury severity and outcome measures in children with traumatic brain injury. AB - The aim of this study was to relate discharge King's Outcome Scale for Childhood Head Injury (KOSCHI) category to injury severity and detailed outcome measures obtained in the first year post-traumatic brain injury (TBI). We used a prospective cohort study. Eighty-one children with TBI were studied: 29 had severe, 15 moderate, and 37 mild TBI. The male:female ratio was 1.8:1. The mean age was 11 years 10 months (SD 3.6, range 5-16y). Discharge KOSCHI categories were good (n=34), moderate (n=39), severe (n=6), and unclassifiable (n=2). KOSCHI category correlated strongly with admission Glasgow Coma Score, length of hospital stay, and post-traumatic amnesia. It also correlated significantly with Verbal IQ and Performance IQ (Wechsler); measures of attention; health status (Health Utilities Index [HUI]); health-related quality of life (Pediatric Quality of Life Inventory [PedsQL]); depressive symptoms (Birleson Depression Scale) assessed within 3 months postTBI; and with Verbal IQ, selective attention (map mission), and HUI and PedsQL domains assessed at least 6 months post-TBI discharge. KOSCHI did not correlate with behaviour or executive function. We conclude that the KOSCHI scored at hospital discharge correlates with severity of injury and some cognitive, health status, and HRQL outcomes early after TBI. It is not helpful at predicting later difficulties, or behavioural and emotional problems. PMID- 18422681 TI - Speech, language, and cognition in preschool children with epilepsy. AB - We studied expressive and receptive language, oral motor ability, attention, memory, and intelligence in 20 6-year-old children with epilepsy (14 females, six males; mean age 6y 5mo, range 6y-6y 11mo) without learning disability, cerebral palsy (CP), and/or autism, and in 30 reference children without epilepsy (18 females, 12 males; mean age 6y 5mo, range 6y-6y 11mo). Ten children had partial, six primarily generalized, and four unclassified epilepsy. Fourteen were having monotherapy and six were taking two or more antiepileptic drugs; 13 children were free from seizures 3 months before the assessment. Results show no statistically significant difference between the groups concerning Verbal IQ, expressive and receptive grammar, and receptive vocabulary. The children with epilepsy had a significantly lower Performance IQ and lower scores in tests of oral motor ability, articulation, emerging literacy, auditory attention, short-term memory, and rapid word retrieval. Parent ratings revealed no significant difference in communicative ability. Polytherapy and early onset of epilepsy influenced some results. Preschool children with epilepsy without learning disability, CP, and/or autism may have receptive verbal ability within the normal range but visuoperceptual, auditory attentional, and speech-language difficulties that could affect school achievement. Careful testing of children with epilepsy who appear to be functioning within the normal range is needed because this may reveal specific impairments that require appropriate professional input. PMID- 18422683 TI - Practical guide of live imaging for developmental biologists. AB - Time-lapse imaging of fluorescent proteins in living cells has become an indispensable tool in biological sciences. However, its application at the organismal level still faces a number of obstacles, such as large specimen sizes preventing illumination of internal tissues, high background fluorescence and uncontrollable movement of target tissues or embryos. Here we describe our solutions for these issues to obtain 4-D fluorescent images from living Drosophila embryos using confocal microscopes. A computational procedure that detects and corrects the shift of moving objects to virtually stabilize them in time-lapse movies (iSEMS) is presented. We discuss the importance of postimaging treatment of raw image stacks for the discovery of novel phenotypes that have previously escaped attention from the analyses of fixed specimens. PMID- 18422682 TI - Intrathecal baclofen therapy in children with intractable spastic cerebral palsy: a cost-effectiveness analysis. AB - In a Dutch national study, we recently established the effectiveness and safety of continuous intrathecal baclofen infusion (CITB) in children with intractable spastic cerebral palsy (CP). Because prospective studies on the cost effectiveness of CITB in children with spastic CP are lacking, we conducted a cost-effectiveness analysis alongside our prospective national study. We compared the costs and health effects of CITB with those of standard treatment only, from the health care perspective for a 1-year period. Health effects were expressed in terms of a visual analogue scale for individual problems and quality-adjusted life years (QALYs). We included eight females and seven males, aged between 7 and 17 years (mean age 13y 8mo [SD 3y]). Eleven children had spastic CP and four had spastic-dyskinetic CP. One child was clsssified on the Gross Motor Function Classification System at Level III, two at Level IV, and 12 at Level V. CITB was more effective and more costly than standard treatment only. Gaining one QALY cost on average 32,737 euros. We conclude that based on the threshold-willingness to pay for one QALY in the Netherlands (80,000 euros), our results confirm the cost-effectiveness of CITB for carefully selected children with intractable spastic CP. PMID- 18422684 TI - Enhancer analysis by chicken embryo electroporation with aid of genome comparison. AB - The identification of the enhancers associated with each developmentally regulated gene is a first step to clarify the regulatory mechanisms underlying embryogenesis. The electroporation technique using chicken embryo is a powerful tool to identify such enhancers. The technique enables us to survey a large genomic region and to analyze the enhancers in great detail. Comparison of the genomic sequences of the chicken and other vertebrate species identifies conserved non-coding sequence blocks to which the functionally identified enhancers often correspond. In this review, I describe in detail the methods to analyze the enhancers using the chicken embryo electroporation and genome comparison. PMID- 18422685 TI - Generation of transgenic medaka using modified bacterial artificial chromosome. AB - The availability of bacterial artificial chromosome (BAC) offers a good genomic platform for a targeted integration of an exogenous gene by a homologous recombination system in Escherichia coli. In combination with microinjection technology, this system allows for the analysis of various aspects of biological phenomena occurring in vivo using Japanese medaka fish (Oryzias latipes). Here we describe a streamlined procedure for selecting BAC clones based on the medaka University of Tokyo genome browser (UTGB), followed by rapid modification with enhanced green fluorescent protein (EGFP) or DsRed fragments for transgenic analysis in medaka. Experimental procedures for BAC DNA preparation, microinjection of medaka embryos and screening of resulting transgenic medaka carrying EGFP/DsRed modified BAC clones are also described. PMID- 18422686 TI - Single cell electroporation method for axon tracing in cultured slices. AB - Axon tracing is an essential method to reveal neuronal connection patterns and characteristics of growing axons during development. Here we introduce an electroporation-based gene transfer technique with a fluorescent protein (FP), which enables us to observe a small number of axons. The result also demonstrates that dynamics of axon behavior can be followed for more than a week. Thus, such labeling with FP is a powerful tool for axon tracing. PMID- 18422687 TI - A single lesion showing features of pigmented eccrine poroma and poroid hidradenoma. AB - Poroid hidradenoma (PH) is a variant of poroma. This entity was defined by Abenoza and Ackerman in 1990. This neoplasm shows architectural characteristics of hidradenoma (tumor cells confined entirely within the dermis in both solid and cystic components) and cytologic characteristics of poroid neoplasm (poroid and cuticular cells, the latter showing ductal differentiation). We herein document a case of single poroid lesion with the features of both eccrine poroma and PH. The patient was a 55-year-old woman with a pigmented nodular lesion on her upper back for 7 years. The histopathologic features of the lesion were consistent with those of eccrine poroma and PH. Unlike most eccrine poromas, this case was pigmented, clinically and microscopically. PMID- 18422688 TI - Pleomorphic angiomatoid fibrous histiocytoma: a case confirmed by fluorescence in situ hybridization analysis for EWSR1 rearrangement. AB - Angiomatoid fibrous histiocytoma (AFH) is a neoplasm of uncertain histogenesis with intermediate malignant potential. It occurs in superficial soft tissues in any age group and at any body site, although the typical example occurs on extremities in children/young adults. The neoplasm is usually composed of a bland histiocytoid proliferation of cells with eosinophilic cytoplasm forming a syncytium, often surrounded by a dense fibrous pseudocapsule and lymphoid infiltrate, with abundant intralesional hemorrhage forming blood-filled spaces. AFH may show striking pleomorphism and mitotic activity, and such cases can lead to confusion with other atypical mesenchymal lesions, including pleomorphic sarcomas. Recent findings have shown that a majority of cases of AFH have translocations involving the EWSR1 or FUS genes. We present a diagnostically challenging case of AFH with pleomorphic features and minimal lymphoid and angiomatoid changes involving the superficial subcutis and deep dermis on the scalp of an 8-year-old boy. "Areas with typical AFH morphology were identified and the diagnosis confirmed with identification of an EWSR1..." rearrangement detected by fluorescence in situ hybridization. Pleomorphic AFH should be included in the differential diagnosis of atypical mesenchymal tumors of skin and superficial subcutis and molecular testing may prove helpful in this regard. PMID- 18422689 TI - Metastatic myoepithelial carcinoma in a child. AB - Cutaneous myoepithelial tumors are rare entities, with few reported malignant variants in the literature. The majority of these tumors are reported in the head and neck region of the adult population, with few examples in the literature arising in young patients. We present a case of myoepithelial carcinoma in a 13 year-old girl, with documented metastatic disease. Reproducible predictors of malignant behavior have yet to be clarified. PMID- 18422690 TI - Cutaneous histopathological findings of Aicardi-Goutieres syndrome, overlap with chilblain lupus. AB - We report a 2-year-old girl with developmental delay who, from the age of 1 year, developed perniotic lesions of the hands and feet initially diagnosed as chilblain lupus. Histological examination showed features of epidermal necrosis with intraepidermal bulla formation, interface dermatitis, lymphocytic vasculitis with fibrinoid necrosis and thrombi formation, both superficial and deep dermal lymphocytic infiltrate, lymphocytic eccrine hidradenitis and absence of marked dermal edema. Subsequent investigations suggested a clinical diagnosis of Aicardi Goutieres syndrome (AGS), a rare genetic leukoencephalopathy. Recently, both AGS and familial chilblain lupus, an autosomal dominant form of systemic lupus erythematosus (SLE), have been shown to be allelic thus suggesting a common pathogenic basis. In addition, a phenotypic overlap is apparent between SLE and AGS. To our knowledge, this is the first comprehensive dermatopathological report of the cutaneous lesions seen in AGS, and our paper highlights the importance of considering AGS in the differential diagnosis of perniosis and chilblain lupus. PMID- 18422691 TI - Genital herpes masquerading as a cutaneous T-cell lymphoma: a report of two cases. AB - Genital herpes simplex virus (HSV) infection is usually a straightforward clinical diagnosis, rarely requiring histological confirmation. We report two cases of immunosuppressed patients in which the clinical and pathological features were initially suspicious for cutaneous lymphoma with a T-cell clone detected in one case. A diagnosis of HSV infection was eventually made on the basis of histological features and confirmed with immunohistochemistry. PMID- 18422692 TI - Malignant cutaneous glomus tumor presenting as a rapidly growing leg mass in a pregnant woman. AB - A 21-year-old pregnant woman presented with a rapidly growing >2 cm nodule on her right leg, involving dermis and subcutaneous tissue. Histologically, the tumor was composed of sheets and nests of neoplastic cells with variable cytomorphology, including typical round to ovoid glomus cells with clear cytoplasm and well-defined cell borders, small cells and spindle cells. Numerous medium to large vessels were present throughout the tumor. Moderate- to high cellularity, nuclear atypia and frequent mitotic figures (42 MF/50 High power field (HPF)) were noted. Immunohistochemistry showed cytoplasmic and membranous expression of actin (HHF-35) and membranous expression of type IV collagen. The histologic features and immunoprofile were consistent with the diagnosis of malignant glomus tumor, a rare soft tissue neoplasm that typically arises on the extremities. Histologic features that infer malignancy in glomus tumors include the combination of large size (>2 cm) and deep location, or atypical mitotic figures, or moderate to severe cytologic atypia with high mitotic activity (>5 mitoses /50 HPF). Although our case was superficially located, the nuclear atypia and mitotic rate, as well as the large size, fulfilled the criteria for a malignant glomus tumor. PMID- 18422693 TI - The challenge of dermatopathological diagnosis of composite tissue allograft rejection: a review. AB - Advances in immunosuppressive treatments and microsurgical techniques have rendered allotransplantation of composite tissues (i.e. heterogeneous, non-organ tissues) possible in humans. Most of these allografts (hands, face and abdominal wall) contain skin that may be the target of rejection, the diagnosis of which relies mainly on clinicopathological monitoring of the skin. Rejection of allografted skin manifests with changes that are characteristic but not very specific. Although composite tissue allografts are still in their infancy, they have opened a new era in the field of transplantation surgery and pathology, so that (dermato) pathologists may occasionally be faced with the challenge of diagnosing skin rejection of a composite tissue allograft. The diagnostic difficulties that may be encountered in the pathological evaluation of skin in this setting are discussed in this review. PMID- 18422694 TI - Leukemia cutis presenting as localized cutaneous hyperpigmentation. AB - The usual clinical presentations of leukemia cutis include solitary infiltrated erythematous or violaceous plaques or nodules and multiple localized or generalized papules. On the other hand, cutaneous hyperpigmentation is a frequent finding in patients with malignancies, most of the cases because of chemotherapy or other drugs that the patient is taking. We present a case of cutaneous hyperpigmentation as the presenting sign of leukemia cutis. A 61-year-old male presented with cutaneous hyperpigmentation, which had appeared during the last chemotherapy cycle for treatment for biphenotypic leukemia. Cutaneous lesions consisted of bluish to brownish irregular well-defined discoloration of the skin involving the upper part of the trunk and the temporal regions of the forehead. The patient was asymptomatic and the skin was not infiltrated at all. However, histopathologic study showed nodular infiltrates involving the full-thickness of the dermis and destroying pre-existing adnexa. This infiltrate was composed of atypical basophilic cells with large hyperchromatic nuclei and scant cytoplasm. Immunohistochemical studies showed intense immunoexpression for CD43, CD68, CD45RO and myeloperoxidase within these cells. A diagnosis of biphenotypic leukemia cutis was established. In our review of the literature we have not found any report of cutaneous hyperpigmentation as the presenting manifestation of leukemia cutis. PMID- 18422695 TI - Chlorhexidine with an anti discoloration system after periodontal flap surgery: a cross-over, randomized, triple-blind clinical trial. AB - The use of chlorhexidine (CHX) has been recommended for a number of clinical applications including plaque control in the post-operative period. However, the use of CHX is burdened by some side effects that could affect the compliance of the patient. The aim of this clinical trial was to evaluate the side effects, the staining in particular, the patient acceptance, and the efficacy of a 0.2% CHX mouthwash containing an anti discoloration system (ADS) compared with a 0.2% CHX alone, after periodontal flap surgery. MATERIAL AND METHODS: This single-centre, cross-over, triple-blind randomized clinical trial was carried out on 48 consecutive patients. After periodontal flap surgery, the patients were prescribed to rinse two times per day for 1 min for 1 week with 10 ml of test or control CHX, contained in anonymous bottles coded K or M and assigned randomly. No brushing and interdental cleaning of the surgical area was allowed. At week 1, after suture removal, patients received full-mouth prophylaxis and were given a second anonymous bottle, reversing the products, with the same instructions as at baseline. Patients resumed tooth-brushing but not interdental cleaning. At the end of week 2, prophylaxis was repeated, mouth rinsing was discontinued and patients resumed normal oral hygiene. At weeks 1 and 2, the following variables were recorded: presence of pigmentation, gingival parameters at the surgically treated sites (gingival inflammation, tissue inflammation around the sutures, gingival swelling and presence of granulation tissue), patient perception and acceptance of the 2 mouthwashes. RESULTS: Forty-seven patients completed the study. The difference between treatments related to gingival variables was not statistically significant. The test CHX caused consistently less pigmentations than the control CHX in all the evaluated areas of the dental surfaces (odds ratio (OR)=0.083 p<0.0001 in the incisal area, OR=0.036 p<0.0001 in the approximal area and OR=0.065 p<0.0001 in the gingival area). The CHX ADS was found to be more tolerated by patients than the control mouthwash and to cause less food alteration, less alterations to the perception of salt and to be less irritant for the oral tissues. CONCLUSIONS: (1) CHX ADS caused less pigmentation, was burdened by less side effects and was more agreeable than the control CHX; (2) CHX ADS was as effective as CHX without ADS in reducing gingival signs of inflammation in the post-surgical early healing phase; (3) the use of CHX ADS could be of value in treatment protocols in which the patient compliance with a CHX mouthwash prescription is relevant. PMID- 18422696 TI - Ridge alterations following immediate implant placement in the dog: flap versus flapless surgery. AB - OBJECTIVE: To assess the healing process after flap or flapless surgery in immediate implant placement. MATERIAL AND METHODS: This study was carried out on five Beagle dogs. Four implants were placed in the lower jaw in each dog immediately after tooth extraction. Flap surgery was performed before the extraction on one side (control), and flapless on the contrary (test). After 3 months of healing, the dogs were sacrificed and prepared for histological analysis. RESULTS: Ten implants were placed in each group. Two failed (one of each group). The percentage of bone-implant contact was very similar in both groups: 64.8% and 65.1% for the flap and the flapless group, respectively. The difference between the mean distance from the peri-implant mucosa margin to the first bone-implant contact at the buccal aspect was statistically significant between both groups (3.02 mm. flapless and 3.69 mm. flap group). The mean first bone-implant contact at the buccal aspect was located in relation to the sand blasted and acid-etched level at 0.82 mm for the flapless group and 1.33 mm for the flap group. This difference was not statistically significant. CONCLUSION: Flapless immediate implant surgery produces a significant reduction in the vestibular biologic width and a minor reduction in buccal bone plate resorption. PMID- 18422697 TI - Mesenchymal stem cells and platelet-rich plasma enhance bone formation in sinus grafting: a histomorphometric study in minipigs. AB - OBJECTIVES: Autologous, allogenic, and alloplastic materials for sinus augmentation have specific drawbacks, which has stimulated an ongoing search for new materials and tissue-engineering constructs. We investigated whether mesenchymal stem cells (MSCs) and platelet-rich plasma (PRP) seeded on a fluorohydroxyapatite (FH) scaffold can improve bone formation and bone-to-implant contact (BIC) in maxillary sinus grafting. MATERIAL AND METHODS: Bilateral sinus augmentation procedures were performed in eight minipigs. MSCs, PRP, and FH scaffold (test site) or FH alone (control site) were grafted in each maxillary sinus. Distal to the osteotomy, one dental implant per sinus was placed in the grafting material through the facial sinus wall. The animals were killed 3 months after grafting, and block sections of the implant sites were harvested and prepared for histomorphometric analysis. RESULTS: After 12 weeks, a significant increase in bone formation occurred in the test sites compared with the control sites (42.51%versus 18.98%; p=0.001). In addition, BIC was significantly greater in the test sites compared with the control sites in the regenerated area (23.71%versus 6.63%; p=0.028). CONCLUSIONS: These findings show that sinus augmentation with MSCs-PRP, combined with FH may enhance bone formation and osseointegration of dental implants compared with FH alone in minipigs. PMID- 18422698 TI - Java project on periodontal diseases: serotype distribution of Aggregatibacter actinomycetemcomitans and serotype dynamics over an 8-year period. AB - OBJECTIVE: To investigate the serotype distribution and stability of Aggregatibacter actinomycetemcomitans over an 8-year period in untreated Indonesian subjects. MATERIAL AND METHODS: Clinical periodontal status and the presence of A. actinomycetemcomitans were established in 1994 and 2002 in 107 subjects from an Indonesian tea estate deprived from dental care. On an average, 3.6 isolates per patient were subcultured and serotyped using specific PCR reactions. RESULTS: In 1994, the predominant serotype was b (53.7%), whereas a and c occurred in 17.1% and 14.6% of the subjects, respectively. In 2002, a reduction in serotypes a (7.5%) and b (30.2%) occurred. Serotypes c and e increased in prevalence from 14.6% to 35.8% and 2.4% to 9.4%, respectively. Multiple serotypes were found in 12.2% in 1994 and 17% in 2002. From 24 subjects who were positive at both time points, 14 (58.3%) had the same serotype, whereas in 10 subjects (41.7%), a different serotype was found. Mean clinical attachment loss had increased from 0.74 mm in 1994 to 1.96 mm in 2002 but could not be related to subgingival presence of A. actinomycetemcomitans. CONCLUSIONS: A. actinomycetemcomitans serotypes distribution in Indonesian young adults shifts from predominantly serotype b to a more equal prevalence of serotypes b and c. This shift suggests an opportunistic character of A. actinomycetemcomitans. PMID- 18422699 TI - Beta-catenin signalling in mesenchymal islet-derived precursor cells. AB - OBJECTIVES: Previously, we characterized human islet-derived precursor cells (hIPCs) as mesenchymal stem cells that migrate out from islets in vitro and can differentiate into functional islet-like structures following proliferative expansion. Here, we investigate the role of beta-catenin signalling in derivation and proliferation of hIPCs. MATERIALS AND METHODS: Localization of beta-catenin was performed using confocal microscopy. Expression levels of beta-catenin target genes were measured by quantitative real-time polymerase chain reaction. Loss-of function studies were performed using specific short interfering RNAs. RESULTS: Immunostaining of islet outgrowths revealed translocation of beta-catenin from plasma membranes in intact islets to the nucleus in cells migrating out. There were no nuclear beta-catenin-positive cells in intact islets whereas between 35% and 70% of cells in established hIPC cultures exhibited nuclear beta-catenin. Transcripts for beta-catenin target genes were increased in hIPCs compared to those in islets. Beta-catenin translocated to the cell membrane when hIPCs formed epithelial cell clusters. In proliferating hIPCs, there was a strong correlation between markers of proliferation and nuclear beta-catenin. Treatment of hIPCs with the glycogen synthase kinase-3beta inhibitor (2'Z,3'E)-6-Bromoindirubin-3' oxime increased intracellular beta-catenin but reduced nuclear beta-catenin, and was associated with reduced cell proliferation. Finally, knockdown of beta catenin decreased beta-catenin target gene expression and hIPC proliferation. CONCLUSIONS: These results support a functional role for beta-catenin during proliferation of hIPCs and suggest that activated beta-catenin signalling may also be important during hIPC derivation from islets. PMID- 18422700 TI - Triiodothyronine stimulates hepatocyte proliferation in two models of impaired liver regeneration. AB - OBJECTIVES: Liver regeneration is attenuated in old age and is substantially slower after 90% than after 70% partial hepatectomy (PH). We have previously demonstrated that the proliferative response to a primary mitogen is intact in aged mice, indicating that impaired liver regeneration is not due to loss of proliferative capacity. Here, we have investigated whether mitogenic effects of triiodothyronine (T3) could reverse the impaired regeneration of ageing or 90% hepatectomy, in the rat. MATERIALS AND METHODS: T3 (20 microg/100 g body weight) was administered to 14-month-old rats subjected to 70% PH or to young rats subjected to 90% PH. Cell-proliferative capacity was determined by bromodeoxyuridine incorporation and microscopy and changes of cell cycle-related proteins were analysed by Western blot analysis. RESULTS: Treatment of old intact rats with T3 increased cyclin D(1) expression that was followed by an enhanced proliferative response, the labelling index (LI), being 7.8% versus 1.3% of controls. T3 given before 70% PH stimulated regenerative response (LI was 10.8% versus 2.28%), and expression of cyclin D(1) and proliferating cell nuclear antigen (PCNA) 24 h after PH. Pre-treatment with T3 also improved the regenerative response of the liver after 90% hepatectomy (LI was 27.9% versus 14.2%). CONCLUSIONS: These findings show in principle that mitogen-induced hyperplasia could be applied to human therapy in patients with reduced regenerative capacity or massive loss of hepatocytes. PMID- 18422701 TI - Human adipose tissue stromal vascular fraction cells differentiate depending on distinct types of media. AB - OBJECTIVES: Angiogenesis, the process of formation of blood vessels, is essential for many physiological as well as pathological processes. It has been shown that human adipose tissue contains a population of non-characterized cells, called stromal-vascular fraction (SVF) cells, which are able to differentiate into several lineages. The aim of this study was to determine conditions for promoting differentiation of human adipose tissue progenitors towards endothelial cells, as well as to show that SVF cells cooperate with differentiated endothelium in capillary network formation. MATERIALS AND METHODS: Stromal vascular fraction cells were isolated according to modified Hauner's method and after adaptation they were cultured in pro-angiogenic or pro-adipogenic medium. Cells were characterized by presence of surface antigens by flow cytometry, and by expression of genes characteristic for endothelial cells or for adipocytes, quantitative real-time polymerase chain reaction. A number of tests were performed to verify their differentiation. RESULTS: Differentiation of human SVF cells towards endothelium was stimulated by the presence of serum and absence of adipogenic factors, documented by the pattern of gene expression as well as different functional in vitro assays. SVF cells were found to work together with human umbilical vein endothelial cells to form capillary networks. CONCLUSIONS: Here, we show that differentiation of SVF cells to endothelial cells or adipocyte like cells depended on the medium used. Our work provides a clear model for analysing the differentiation capacity of SVF cells. PMID- 18422702 TI - Ratio of Wnt3a to BMP4 doses is critical to their synergistic effects on proliferation of differentiating mouse embryonic stem cells. AB - OBJECTIVES: To investigate potential interactions between bone morphogenetic protein (BMP) and Wnt signalling on differentiating mouse embryonic stem cells (mESC). MATERIALS AND METHODS: Mouse embryonic stem cells were cultured with differing combinations of Wnt3a, BMP4 and inhibitors of Wnt, BMP, PI-3K (phosphoinositide 3-kinase), p38, ERK1/2 (extracellular signal-regulated kinase 1/2) and JNK (c-Jun N-terminal kinase) pathways. RESULTS: We found that Wnt3a synergized with BMP4 to promote mESC proliferation. Furthermore, the relative ratio of Wnt3a to BMP4 doses was critical to their synergistic effects, which could be abolished by using Dkk-1, noggin or the inhibitors of PI-3K, p38, ERK1/2 and JNK pathways. We also demonstrated that combination of Wnt3a and BMP4 could suppress ectodermal differentiation of mESCs. Moreover, inhibitors of PI-3K, p38, ERK1/2 and JNK pathways could negate this effect. CONCLUSION: Relative ratio of Wnt3a to BMP4 doses is critical to their synergistic effect on differentiating mESC proliferation, which may work through PI-3K, p38, ERK1/2 and JNK pathways. PMID- 18422703 TI - Novel role for polycystin-1 in modulating cell proliferation through calcium oscillations in kidney cells. AB - OBJECTIVES: Polycystin-1 (PC1), a signalling receptor regulating Ca(2+)-permeable cation channels, is mutated in autosomal dominant polycystic kidney disease, which is typically characterized by increased cell proliferation. However, the precise mechanisms by which PC1 functions on Ca(2+) homeostasis, signalling and cell proliferation remain unclear. Here, we investigated the possible role of PC1 as a modulator of non-capacitative Ca(2+) entry (NCCE) and Ca(2+) oscillations, with downstream effects on cell proliferation. RESULTS AND DISCUSSION: By employing RNA interference, we show that depletion of endogenous PC1 in HEK293 cells leads to an increase in serum-induced Ca(2+) oscillations, triggering nuclear factor of activated T cell activation and leading to cell cycle progression. Consistently, Ca(2+) oscillations and cell proliferation are increased in PC1-mutated kidney cystic cell lines, but both abnormal features are reduced in cells that exogenously express PC1. Notably, blockers of the NCCE pathway, but not of the CCE, blunt abnormal oscillation and cell proliferation. Our study therefore provides the first demonstration that PC1 modulates Ca(2+) oscillations and a molecular mechanism to explain the association between abnormal Ca(2+) homeostasis and cell proliferation in autosomal dominant polycystic kidney disease. PMID- 18422704 TI - Occlusal caries detection in permanent molars according to WHO basic methods, ICDAS II and laser fluorescence measurements. AB - This epidemiological study aimed to compare the diagnostic outcome of the WHO criteria, ICDAS II criteria, laser fluorescence measurements, presence of plaque and roughness as activity scores on occlusal fissures and buccal/palatal pits of the first permanent molars. The study involved 311 children between 8 and 12 years of age from the Ennepe-Ruhr District in North Rhine-Westphalia, Germany. The surface-related caries status was registered according to the WHO basic method criteria (1997). Additionally, pit and fissure sealants, the ICDAS II visual criteria, the DIAGNOdent reading, plaque retention and surface roughness were documented. Caries experience was 1.0 (+/-2.5) DMFS. About 70% of the examined students had no obvious dentin caries in the permanent dentition (DMFS = 0). Sealants were registered on 1.4 (+/-1.7) occlusal fissures and 0.4 (+/-0.9) palatal/buccal pits. Noncavitated caries lesions were recorded as ICDAS II score 1-4 on 1.8 (+/-1.6) fissures and 1.5 (+/-1.4) pits. The comparison of the diagnostic methods suggests a relationship between higher ICDAS II scores/DIAGNOdent values and a proportional increase in the occurrence of plaque as well as in the number of rough surfaces. In conclusion, this study showed the diagnostic potential of the ICDAS II criteria in comparison to the traditional WHO criteria by means of the noncavitated caries lesions additionally detected. The DIAGNOdent use in field studies that already apply detailed visual criteria seems to bring limited additional information. While the presence of plaque provides information for the caries activity assessment more work is required to provide information about the contribution of surface roughness. PMID- 18422705 TI - Refining exposure definitions for studies of periodontal disease and systemic disease associations. AB - BACKGROUND: Substantial variation exists in reported associations between periodontal infections and cardiovascular disease. Imprecise periodontal exposure definitions are possible contributors to this variability. We studied appropriate exposure definitions for studying associations between clinical periodontal disease (PD) and systemic disease. METHODS: Data originate from men and women aged 20-79 enrolled in the Study of Health in Pomerania (SHIP) from 1997-2001. Age and sex-adjusted correlation analysis identified PD definitions with the highest cross-sectional associations with three subclinical markers of systemic disease: plasma fibrinogen (n = 3481), serum hemoglobin A1c (HbA1c) (n = 3480), and common carotid artery intima-media thickness (c-IMT) (n = 1745, age > or = 45). RESULTS: In men and women, percent of sites with attachment loss (AL) > or =6 mm and tooth loss both revealed the highest correlation with HbA1c (rho = 0.11; several other definitions related similarly), while the strongest fibrinogen correlation was observed with percent of sites with pocket depth > or =3 mm (rho = 0.19). Findings for c-IMT among men were strongest for percent of sites with AL > or =6 mm (rho = 0.14; several other definitions related similarly) while among women, percent of sites with pocket depth > or =5 or 6 mm had the highest observed correlation (rho = 0.13). CONCLUSIONS: A range of near optimal definitions varied according to gender and whether the systemic disease marker reflected an acute or chronic situation. Pocket depth was more strongly correlated with the acute marker fibrinogen while attachment and tooth loss tended to be more strongly correlated with the chronic markers, HbA1c, and c-IMT. These findings can be useful in designing future studies investigating the association between PD and systemic disease. PMID- 18422706 TI - Testing responsiveness to change for the early childhood oral health impact scale (ECOHIS). AB - OBJECTIVES: The Early Childhood Oral Health Impact Scale (ECOHIS) is a recently developed oral health-related quality of life instrument designed to assess the impact of oral health problems in 0-5-year-old children. It has previously been validated as discriminative instrument. The goal of this study was to investigate the responsiveness to change of the ECOHIS. METHODS: Data were collected from a convenience sample of 101 parents of 0-5-year-old children attending a hospital dental clinic for dental treatment. The ECOHIS was completed by parents prior to dental treatment and 2 weeks later. Subjects were also asked a global transition judgement concerning change between the second and first completion of the ECOHIS instrument. Responsiveness to change of the ECOHIS was analysed through: (i) a comparison of ECOHIS change scores with a global transition judgment by study subjects; (ii) an assessment of the statistical significance of within-group change in scores over time for groups reporting improvement, stability and deterioration; (iii) an estimation of the ECOHIS's sensitivity; and (iv) an investigation the effect size of the ECOHIS. RESULTS: Of the 101 subjects recruited, 94 had full datasets. Their data were used for the analyses reported in this paper. Pre- and post-treatment distributions of ECOHIS scores were strongly distributed towards no oral health impacts. Among the 94 subjects, 51.1% reported improvement, 42.6% reported no change and 6.4% reported deterioration following treatment, using the global transition judgement. The mean ECOHIS change scores for these three groups were -0/9, +0.7 and +6.5 respectively, although none of the within-group changes were statistically significant. The effect size for those reporting improvement was small (0.15) but for those reporting deterioration was moderate-to-large (0.69). Sensitivity ranged from 0.61-0.79 depending on the size of the cut-off point, with a change of 3 points demonstrating the best sensitivity to false positive ratio (0.79 versus 0.41 respectively). CONCLUSION: In this sample with low levels of problems, the ECOHIS has demonstrated some limited ability to respond to change. Further work in a larger sample with higher levels of problems is needed to investigate the instrument's ability to respond to change when it has occurred. PMID- 18422707 TI - Oral health equality during 30 years in Norway. AB - OBJECTIVES: To study the relationship between income and edentulousness and having a functional dentition from 1975 to 2002, and to examine whether or not the findings can be characterized as a social gradient. METHODS: Four datasets were collected by personal interviews and precoded questionnaires by Statistics Norway in 1975, 1985, 1995 and 2002. The datasets were representative of the non institutionalized adult population in Norway. Each year, the sampled persons were distributed into five income quintiles. Four effects were examined of the relationship between income quintiles and edentulousness and functional dentition (1) the absolute equality effect, (2) the period effect, (3) the relative equality effect, and (4) the gradient effect. RESULTS: The main finding is that in absolute terms oral health is more equally distributed in 2002 than in 1975, and the lowest income groups benefited the most in oral health. Among the elderly, however, having a functional dentition was less equally distributed in 2002. The relative differences increased for the oldest for each new birth cohort; thus, the chances of being edentulous was 7.5 times higher in the lowest income group versus the highest group in 2002, whereas the chances were only two times higher in 1972. Having lost all natural teeth was infrequent in 2002, and inequality wiped out in the population below > or = 60 years. A small social gradient was still in 2002. CONCLUSIONS: The condition of edentulousness is a result of accumulated incidences of dental diseases and fragmented access to dental care. Economic barriers and unavailability of dental care postponed necessary restorative dental care and resulted in more drastic treatment solutions previously especially in the rural areas in Norway. PMID- 18422708 TI - Prevalence of hyposalivation in relation to general health, body mass index and remaining teeth in different age groups of adults. AB - OBJECTIVES: Several studies have been conducted on the prevalence of hyposalivation in older adults but no population-based studies in younger adults. Therefore, our aims were to determine the prevalence of very low and low unstimulated (UWSFR) and stimulated (SWSFR) whole salivary flow rates in different age groups between 20 and 69 years, and to analyse the relationship between hyposalivation, subjective oral dryness and predictors of reduced flow rate. METHODS: A randomized and stratified cross-sectional study including 1427 dental patients was conducted. UWSFR and SWSFR were measured, numbers of remaining teeth recorded and a questionnaire answered regarding subjective oral dryness, general diseases, use of drugs, body mass index (BMI) and use of tobacco. RESULTS: The prevalence of very low (<0.1 ml/min) and low (0.10-0.19 ml/min) UWSFR was similar for different age groups up to 50 years, ranging between 10.9-17.8% and 17.3-22.7%, respectively. The prevalence of very low UWSFR was significantly higher for women aged 50-69 years than for younger women. For men, prevalence of very low UWSFR was higher at 60-69 years. The prevalence of very low (<0.7 ml/min) and low (0.70 - 0.99 ml/min) SWSFR was between 0-5.5% and 0.8-8.2%, respectively, for the different age groups 20-69 years. Multiple logistic regression revealed that age above 50 years, female gender, having fewer than 20 teeth, and taking xerogenic drugs significantly increased the risk of very low UWSFR. For very low SWSFR, only having fewer than 20 teeth and taking more than two drugs were significant. In the younger individuals (<50 years) only BMI > 25 for very low UWSFR and diagnosed disease for very low SWSFR were found significant. In this younger subset, female gender combined with having fewer than 27 teeth was significant for low UWSFR. CONCLUSIONS: Hyposalivation is prevalent in younger adults, among whom it is associated with diagnosed disease and high BMI, while after age 50 years it is associated with medication. It is also associated with gender and with fewer remaining teeth. PMID- 18422709 TI - Lifetime fluoridation exposure and dental caries experience in a military population. AB - While there is good evidence of caries-preventive benefits of fluoride in drinking water among children and adolescents, there is little information about effectiveness of water fluoridation among adults. OBJECTIVES: To determine whether exposure to fluoride in drinking water is associated with caries experience in Australian Defence Force (ADF) personnel. METHODS: Cross-sectional study of 876 deployable ADF personnel aged 17-56 years. At each person's mandatory annual dental examination, military dentists recorded the number of decayed, missing and filled teeth (DMFT) using visual, tactile and radiographic criteria. Participants also completed a questionnaire, listing residential locations in each year from 1964 to 2003. People were classified into four categories according to the percentage of their lifetime living in places with fluoridated water: <10%, 10% to <50%, 50% to <90% and > or =90%. Mean DMFT was compared among those categories of fluoridation exposure and the association was evaluated statistically using analysis of variance to adjust for age, sex, years of service and rank. RESULTS: Without adjustment for confounders, the mean DMFT (+/-95% confidence interval) was 6.3 +/- 0.8 for <10% fluoridation exposure, 7.8 +/- 0.8 for 10% to <50% exposure, 7.5 +/- 0.7 for 50% to <90% exposure and 4.6 +/ 0.6 for > or =90% exposure (P < 0.01). However, age was inversely associated with mean DMFT and in the <10% exposure group, 91% of people were aged <35 years. Service rank was also significantly associated with both fluoridation exposure and DMFT. After adjustment for all covariates, mean DMFT was 24% lower among people in the two groups with > or =50% exposure compared with the <10% exposure group. CONCLUSIONS: Degree of lifetime exposure to fluoridated drinking water was inversely associated with DMFT in a dose-response manner among this adult military population. PMID- 18422710 TI - Caries increment in children aged 3-5 years in relation to parents' dental attitudes: Oslo, Norway 2002 to 2004. AB - OBJECTIVES: To assess the relationship between parents' dental attitudes and the caries increment in their children from the age of 3 to 5 years. METHODS: Data based on parental questionnaires and dental examinations were collected from children participating in a follow-up study from age 3 years (n = 354) in 2002 to 5 years (n = 304) in 2004. The children were categorized as western-native (WN) and immigrants (IM). The items used were significantly related to caries experience in a multicentre study [Pine et al. (2004)Community Dent Health, vol. 21, pp. 121-30]. The responses to attitudinal items were weighted as positive if they would promote good dental health, and negative if not. Composite attitudinal variables relating to hygiene, diet and indulgence were calculated as a summation of the weighted responses to selected items. Regression analyses (bivariate and multiple) were performed to assess associations during the period between the attitudinal predictors/other control variables and caries increment (Deltad(3 5)mfs). RESULTS: Bivariate logistic regression analyses revealed that 'Attitude to Diet' and 'Parental Indulgence' were clearly related to caries increment. The more exposed children were to negative parental attitudes, the higher the OR. 'Attitude to Diet' also persisted in a multiple logistic regression model, showing a higher OR value than caries experience. 'Immigrant Status' was the most potent predictor of caries increment. Parents were found to be more indulgent among IM than among WN groups. CONCLUSION: Parental dental attitudes are clearly shown to be associated with caries increment in early childhood. The relationship is of such strength that it deserves to be taken into account in future preventive dental strategies. PMID- 18422711 TI - A 2-year community-randomized controlled trial of fluoride varnish to prevent early childhood caries in Aboriginal children. AB - OBJECTIVE: To measure the effectiveness of fluoride varnish (FV) (Duraflor), 5% sodium fluoride, Pharmascience Inc., Montreal, QC, Canada) and caregiver counseling in preventing early childhood caries (ECC) in Aboriginal children in a 2-year community-randomized controlled trial. METHODS: Twenty First Nations communities in the Sioux Lookout Zone (SLZ), Northwest Ontario, Canada were randomized to two study groups. All caregivers received oral health counseling, while children in one group received FV twice per year and the controls received no varnish. A total of 1275, 6 months to 5-year-old children from the SLZ communities were enrolled. In addition, a convenience sample of 150 primarily non Aboriginal children of the same age were recruited from the neighboring community of Thunder Bay and used as comparisons. Longitudinal examinations for the dmft/s indices were conducted by calibrated hygienists in 2003, 2004 and 2005. RESULTS: Aboriginal children living in the SLZ or in Thunder Bay had significantly higher caries prevalence and severity than non-Aboriginal children in Thunder Bay. FV treatment conferred an 18% reduction in the 2-year mean 'net' dmfs increment for Aboriginal children and a 25% reduction for all children, using cluster analysis to adjust for the intra-cluster correlation among children in the same community. Adjusted odds ratio for caries incidence was 1.96 times higher in the controls than in the FV group (95% CI = 1.08-3.56; P = 0.027). For those caries-free at baseline, the number (of children) needed to treat (NNT) equaled 7.4. CONCLUSIONS: Findings support the use of FV at least twice per year, in conjunction with caregiver counseling, to prevent ECC, reduce caries increment and oral health inequalities between young Aboriginal and non-Aboriginal children. PMID- 18422712 TI - The effect of anticoagulant, storage temperature and dilution on cord blood hematology parameters over time. AB - The objective of the study was to determine whether selected hematologic parameters measured on umbilical cord blood samples using an automated hematology analyzer (Sysmex XE-2100) were affected by (i) anticoagulant (the specimens were collected in EDTA vs. sodium heparin), (ii) temperature (the specimens were maintained at 4 degrees C vs. room temperature for up to 72 h) and (iii) 1 : 5 dilution vs. undiluted using the manufacturer's diluting solution. Use of heparin, instead of EDTA, had little effect on the hematologic results (n = 8) except for lower platelet and progenitor cell counts. Results were remarkably stable for 72 h at either room temperature or 4 degrees C except for modest red blood cell swelling at 24 h. Specimens of blood diluted at 1 : 5 had an immediate small, but significant change on white cell count (+13.3%), reticulocyte count ( 11.2%) and reticulocyte hemoglobin content (-19.6%). Diluted samples did not change further over 4 h at room temperature. With a 1 : 5 dilution, analysis of 40 microl of cord blood stored for 3 days at room temperature may provide useful hematologic information with little phlebotomy loss. PMID- 18422713 TI - Biomechanical aspects of primary implant stability: a human cadaver study. AB - BACKGROUND: The quality of bone is an important factor in the successful implant treatment, and it is evident that higher implant failure is more likely in poor quality of bone. The primary stability of oral implants related to resistance to micromotion during healing is influenced by bone quality, surgical technique, and implant design. PURPOSES: The aims of this biomechanical study were to explore the effect of bone quality on initial intraosseous stability of implants, and to determine the correlations between the bone quality and implant stability parameters. MATERIALS AND METHODS: Twenty-four implants (Neoss Ltd., Molnlycke, Sweden) were placed into anterior and posterior regions of three human cadaver mandibles. The bone densities of implant recipient sites were preoperatively determined using computerized tomography (CT) in Hounsfield unit (HU). The maximum insertion torque values were recorded, and primary implant stability measurements were noninvasively performed by means of resonance frequency analysis (RFA). RESULTS: The bone density values ranged from -267 HU to 553 HU. It was found that mean bone density, insertion torque, and RFA values were 113 +/ 270 HU, 41.9 +/- 5 Ncm, and 70 +/- 7 implant stability quotient (ISQ), respectively. Statistically significant correlations were found between bone density and insertion torque values (r = 0.690, p < .001); bone density and ISQ values (r = 0.557, p < .05); and insertion torque and ISQ values (r = 0.853, p < .001). CONCLUSION: CT is a useful tool to assess bone quantity and quality in implant recipient sites, and bone density has a prevailing effect on implant stability at placement. PMID- 18422715 TI - The capsaicin-blink reflex linkage in chronic migraine revisited. PMID- 18422716 TI - Are signs of temporomandibular disorders stable and predictable in adolescents with headache? AB - The aim of the present study was to study changes in signs and symptoms of temporomandibular disorders (TMD) and factors predicting TMD signs in adolescents with and without headache. A population-based sample (n = 212) of 13-year-olds with and without headache was re-examined at the age of 16. The study included a questionnaire, face-to-face interview and somatic examination. In addition, a neurological examination, a muscle evaluation and a stomatognathic examination were performed. Significant changes were seen in TMD signs during the follow-up, but TMD signs at the end of the follow-up could not be predicted by baseline headache, sleeping difficulties, depression or muscle pain. TMD signs at the age of 16 were associated with female gender and muscle pain. We conclude that considerable changes in TMD signs occur in the follow-up of adolescents with and without headache. Headache-related TMD are not predictable in adolescents with and without headache. PMID- 18422717 TI - The incidence and prevalence of cluster headache: a meta-analysis of population based studies. AB - Cluster headache is a trigemino-autonomic cephalgia with a low prevalence. Several population-based studies on its prevalence and incidence have been performed, but with different methodology resulting in different figures. We analysed all available population-based epidemiological studies on cluster headache and compared the data in a meta-analysis. The pooled data showed a lifetime prevalence of 124 per 100,000 [confidence interval (CI) 101, 151] and a 1-year prevalence of 53 per 100,000 (CI 26, 95). The overall sex ratio was 4.3 (male to female), it was higher in chronic cluster headache (15.0) compared with episodic cluster headache (3.8). The ratio of episodic vs. chronic cluster headache was 6.0. Our analysis revealed a relatively stable lifetime prevalence, which suggests that about one in 1000 people suffers from cluster headache, the prevalence being independent of the region of the population study. The sex ratio (male to female) is higher than published in several patient-based epidemiological studies. PMID- 18422719 TI - C1-C2 fluid collection on MRI myelography in patients with spontaneous intracranial hypotension. PMID- 18422720 TI - Acute convexity subarachnoid haemorrhage: a cause of aura-like symptoms in the elderly. AB - Late-life onset aura-like symptoms are not rare and can be potentially misdiagnosed as transient ischaemic attacks. The cause is often obscure. Four cases of recurrent aura-like symptoms are presented in whom acute convexity subarachnoid haemorrhage (SAH) contralateral to symptoms was demonstrated. Three experienced subsequent events or groups of events triggered by recurrent SAH. Negative diffusion-weighted imaging, normal electroencephalography and slow symptom march with complete resolution argued against ischaemic and epileptic causes. Aura-like symptoms in the elderly should be investigated with imaging modalities most sensitive for detecting subarachnoid blood, in particular gradient echo magnetic resonance imaging. PMID- 18422721 TI - A role for wind-up in trigeminal sensory processing: intensity coding of nociceptive stimuli in the rat. AB - Wind-up is a progressive, frequency-dependent increase in the excitability of trigeminal and spinal dorsal horn wide dynamic range (WDR) nociceptive neurons evoked by repetitive stimulation of primary afferent nociceptive C-fibres. The correlate of wind-up in humans is temporal summation, which is an increase in pain perception to repetitive constant nociceptive stimulation. Although wind-up is widely used as a tool for studying the processing of nociceptive information, including central sensitization, its actual role is still unknown. Here, we recorded from trigeminal WDR neurons using in vivo electrophysiological techniques in rats and assessed the wind-up phenomenon in response to stimuli of different intensities and frequencies. First, we found that the amplitude of C evoked responses of WDR neurons to repetitive stimulation increased progressively to reach a peak, then consistently showed a stable or slightly decreasing plateau phase. Only the first phase of this time course fitted in with the wind-up description. Therefore, to assess wind-up, we measured a limited number of initial responses. Second, we showed that wind-up, i.e. the slope of the frequency-dependent increase in the response to C-fibre stimulation, was linearly correlated to the stimulus intensity. Intensities of brief C-fibre inputs were thus coded into frequencies of action potentials by second-order neurons through frequency-dependent potentiation of the evoked responses. Third, wind-up also occurred at stimulation intensities below the threshold for C-evoked responses in WDR neurons, suggesting that wind-up can amplify subthreshold C-fibre inputs to WDR neurons. This might account for the observation that sparse, subliminal, neuronal activity in nociceptors can become painful via central integration of neural responses. Altogether, the present results show that wind-up can provide trigeminal WDR neurons with the capability to encode the intensity of short duration orofacial nociceptive stimuli and to detect subthreshold nociceptive input. Thus, not only may wind-up play a physiological role in trigeminal sensory processing, but its enhancement may also underlie the pathophysiology of chronic orofacial pain conditions. PMID- 18422723 TI - Comparisons between heavily T2-weighted MR and CT myelography studies in two patients with spontaneous intracranial hypotension. AB - Our previous study demonstrated that heavily T2-weighted magnetic resonance (MR) myelography could successfully detect abnormal cerebrospinal fluid (CSF) collections in patients with spontaneous intracranial hypotension (SIH); however, its ability to demonstrate ongoing CSF leakage sites is uncertain. Currently, computed tomographic (CT) myelography is still considered the study of choice for such a purpose. In this study, we compared the results of the two imaging modalities in two patients with SIH. CSF leakages, such as C1-2 extraspinal collections, CSF along nerve root sleeves, and epidural fluid collections were noted on both studies with minor discrepancies. CSF collection along nerve root sleeves demonstrated by MR myelography was the most likely ongoing leakage site in comparison with CT myelography. Targeted epidural blood patches in both patients resulted in complete headache resolution. Non-invasive, non-contrasted and time-saving MR myelography is a promising study for patients with SIH. Further large-scale validation studies are needed. PMID- 18422722 TI - Unilateral photophobia or phonophobia in migraine compared with trigeminal autonomic cephalalgias. AB - Our objective was to compare the presence of self-reported unilateral photophobia or phonophobia, or both, during headache attacks comparing patients with trigeminal autonomic cephalalgias (TACs)--including cluster headache, short lasting unilateral neuralgiform headache attacks with conjunctival injection and tearing (SUNCT) and paroxysmal hemicrania--or hemicrania continua, and other headache types. We conducted a prospective study in patients attending a referral out-patient clinic over 5 months and those admitted for an intramuscular indomethacin test. Two hundred and six patients were included. In episodic migraine patients, two of 54 (4%) reported unilateral photophobia or phonophobia, or both. In chronic migraine patients, six of 48 (13%) complained of unilateral photophobia or phonophobia, or both, whereas none of the 24 patients with medication-overuse headache reported these unilateral symptoms, although these patients all had clinical symptoms suggesting the diagnosis of migraine. Only three of 22 patients (14%) suffering from new daily persistent headache (NDPH) experienced unilateral photophobia or phonophobia. In chronic cluster headache 10 of 21 patients (48%) had unilateral photophobia or phonophobia, or both, and this symptom appeared in four of five patients (80%) with episodic cluster headache. Unilateral photophobia or phonophobia, or both, were reported by six of 11 patients (55%) with hemicrania continua, five of nine (56%) with SUNCT, and four of six (67%) with chronic paroxysmal hemicrania. Unilateral phonophobia or photophobia, or both, are more frequent in TACs and hemicrania continua than in migraine and NDPH. The presence of these unilateral symptoms may be clinically useful in the differential diagnosis of primary headaches. PMID- 18422724 TI - Population-based validation of a German-language self-administered headache questionnaire. AB - We validated a German-language self-administered headache questionnaire for migraine (M), tension-type headache (TTH) and trigeminal autonomic cephalalgia (TAC) in a general population sample of people with headache. Randomly selected subjects (n = 240) diagnosed by the questionnaire as M (n = 60), TTH (n = 60), a combination of M and TTH (M+TTH, n = 60) and TAC (n = 60) were invited for examination by headache specialists. One hundred and ninety-three subjects (80%) were studied. Sensitivity and specificity for M were 0.85 and 0.85, for TTH 0.6 and 0.88, for M+TTH 0.82 and 0.87, respectively. Cohen's kappa was 0.6 (95% confidence interval 0.50, 0.71). Of 45 patients with TAC according to the questionnaire, physicians diagnosed cluster headache in two patients only. We conclude: (i) the questionnaire can be used to diagnose M, TTH and M+TTH, but not TAC; (ii) screening questionnaires for epidemiological research should be validated in a general population sample but not in a tertiary headache clinic. PMID- 18422725 TI - Regional grey matter changes in patients with migraine: a voxel-based morphometry study. AB - We used voxel-based morphometry (VBM) to compare grey matter volume (GMV) between 20 migraine patients (five with aura and 15 without aura) with normal conventional magnetic resonance imaging findings and 33 healthy controls matched for age and sex. A separate analysis was also performed to delineate a possible correlation between the GMV changes and the headache duration or lifetime headache frequency. When compared with controls, migraine patients had significant GMV reductions in the bilateral insula, motor/premotor, prefrontal, cingulate cortex, right posterior parietal cortex, and orbitofrontal cortex (P < 0.001, uncorrected for multiple comparisons at a voxel level; corrected P < 0.05 after small volume corrections). All regions of the GMV changes were negatively correlated with headache duration and lifetime headache frequency (P < 0.05, Pearson's correlation test). We found evidence for structural grey matter changes in patients with migraine. Our findings of progressive GMV reductions in relation to increasing headache duration and increasing headache frequency suggest that repeated migraine attacks over time result in selective damage to several brain regions involved in central pain processing. PMID- 18422726 TI - Genomic and functional analyses of MUTYH in Japanese patients with adenomatous polyposis. AB - The present study was undertaken to elucidate germ line mutations of the base excision repair gene, MUTYH, in Japanese patients with adenomatous polyposis. We screened germ line mutations of adenomatous polyposis coli (APC) gene and MUTYH in 66 Japanese patients with adenomatous polyposis. APC was screened by the protein truncation test, while MUTYH was screened by polymerase chain reaction based single-strand conformation polymorphism and direct sequencing. The nicking assay was applied in order to evaluate the DNA glycosylase activity of the identified MUTYH variant. In this study, Seven MUTYH variants were identified in 16 of 21 APC-negative patients. Q324H mutation was the most frequent mutation, with an allele frequency of 49%. Two patients carried biallelic mutations other than Q324H; a patient had biallelic G272E and A359V mutations, while the other had compound heterozygotes of P18L and G25D mutations. Nicking assay for G272E using the corresponding mouse MUTYH mutant with G257E revealed that G272E is a variant to cause an impaired DNA glycosylase activity. Homozygous MUTYH mutation accounts for approximately 10% of Japanese patients with adenomatous polyposis. G272E may be one of the mutations specific to patients with adenomatous polyposis in East Asia. PMID- 18422727 TI - Immunological efficacy of heat shock protein 60 peptide DiaPep277 therapy in clinical type I diabetes. AB - An immunogenic peptide (p277) from the 60-kDa heat shock protein (hsp60) arrested beta-cell destruction in non-obese diabetic mice. A randomized, double-blind, phase Ib/II clinical trial of DiaPep277 peptide treatment was performed in recent onset type 1 diabetes patients with remaining insulin production. We studied the immunological efficacy of this peptide therapy and correlated this with clinical outcome. Forty-eight C-peptide-positive patients were assigned subcutaneous injections of 0.2, 1.0 or 2.5 mg p277 (n = 12 per dosage) at entry, and 1, 6 and 12 months, or four placebo injections (n = 12). T cell autoimmunity to hsp60, DiaPep277, glutamic acid decarboxylase and tetanus toxoid (recall response control) were assayed by proliferation and cytokine secretion assays (enzyme linked immunospot) at regular intervals until 18 months after the first injection. All treated patients at each dosage of peptide demonstrated an altered immune response to DiaPep277, while the majority of placebo-treated patients remained non-responsive to treatment (P = 0.00001), indicating a 100% efficacy of immunization. Cytokine production in response to therapy was dominated by interleukin (IL)-10. IL-10 production before therapy and decreasing autoantigen specific T cell proliferation were associated with beta-cell preservation. Third party control immune responses were unaffected by therapy. No potentially adverse immunological side effects were noted. DiaPep277 is immunogenic in type 1 diabetic subjects and has immune modulating properties. Immunological monitoring distinguished therapy from placebo treatment and could determine immunological efficacy. Declining or temporary proliferative responses to peptide DiaPep277 treatment may serve as an immunological biomarker for clinical efficacy. PMID- 18422728 TI - Ageing is associated with diminished apoptotic cell clearance in vivo. AB - Ageing leads to immune system dysfunction and the accumulation of autoantibodies. Because the rapid phagocytic clearance of apoptotic cells is required to prevent the development of autoimmunity, we examined the relative clearance of apoptotic material in young and aged mice using two independent assays. First, 2-year-old mice were found to be impaired in their ability to clear apoptotic keratinocytes following ultraviolet irradiation of the skin. Secondly, peritoneal macrophages exposed to apoptotic Jurkat T cells in vivo displayed diminished phagocytic activity in aged mice compared with 8-week-old mice. Consistent with these findings, aged mice exhibited signs of autoimmunity with the appearance of anti nuclear antibodies and increased kidney glomerular size as well as complement deposits within the glomeruli. In vitro assays revealed that the pretreatment of macrophages with the serum from aged mice led to a reduction in their ability to phagocytose apoptotic bodies compared with macrophages treated with serum from young mice. These data show that the ageing process is accompanied by a diminished ability to clear apoptotic debris. This accumulation of apoptotic debris could contribute to immune system dysfunction that occurs in aged organisms. PMID- 18422729 TI - Elevated CCR6+ CD4+ T lymphocytes in tissue compared with blood and induction of CCL20 during the asthmatic late response. AB - CCR6 is expressed by multiple leucocyte subsets, including peripheral blood memory T cells, and mouse models implicate a role for this receptor in diverse inflammatory responses that include allergic airway disorders, inflammatory bowel disease and autoimmune encephalitis. In order to study the role of CCR6 in humans, we have investigated the patterns of CCR6 expression and function on T cells from the peripheral blood, skin, nose and lung, in health and in allergic disease. Results show that CCR6 was expressed consistently on a higher proportion of tissue versus peripheral blood-derived CD4+ T cells (P < 0.01). CCR6 was expressed predominantly on CD4+ compared with CD8+ cells in both blood- and tissue-derived T cells (P < 0.001). The number of cells showing CCR6 expression was not proportionally greater in peripheral blood or nasal mucosal T cells of subjects with symptomatic allergic rhinitis. CCR6+ cells demonstrated enhanced functional responses to CCL20 and CCL20 was increased in bronchoalveolar lavage fluid of asthmatics following endobronchial allergen provocation (P < 0.05). Thus, CCR6 may be important in the regulation of T cell recruitment to tissue and up-regulation of CCL20 expression may contribute to the recruitment and/or retention of effector T cells in allergic asthma. PMID- 18422730 TI - Interleukin 12B gene polymorphism and apparent resistance to hepatitis C virus infection. AB - Cellular immunity with interferon gamma production could have a role in protection from hepatitis C virus (HCV). Interleukin (IL)-12 is a key cytokine in promoting such anti-viral T helper 1 (Th1) responses. We hypothesized that a genetic background able to promote cellular responses may be associated with apparent protection from infection and have investigated the distribution of the functional 1188A/C polymorphism of IL-12B in HCV exposed but uninfected cases. The frequency of the high IL-12-producing C allele was determined by restriction enzyme genotyping in 76 exposed-uninfected individuals and 105 healthy controls. Overall, the C allele was found in 27.6% of exposed-uninfected cases compared with 16.7% of healthy controls [chi(2) = 6.3, P = 0.02, odds ratio (OR) = 1.9, 95% confidence interval (CI) = 1.1-3.2]. CC genotype was found in 10.5% of exposed-uninfected cases compared with 0.9% controls (chi(2) = 9.3, P = 0.01, OR = 12, 95% CI = 1.5-100). Individuals at high risk of HCV infection yet who remain uninfected may be resistant in some way to infection. In our cohort of exposed uninfected cases a genetic background of enhanced IL-12 production was associated with apparent resistance to HCV infection. This lends support to a central role for cellular immune responses in protecting from infection. PMID- 18422731 TI - The profile of gene expression and role of nuclear factor kappa B on glomerular injury in rats with Thy-1 nephritis. AB - Mesangioproliferative glomerulonephritis (MsPGN) is a disease of high incidence in humans. Rats with Thy-1 nephritis (Thy-1 N) are used as an animal model for studying MsPGN. Although several studies have demonstrated that many pathological factors could cause the injury of glomerular mesangial cells (GMCs) in Thy-1 N, changes of profile and the molecular mechanism of the disease (i.e. the role of transcription factors) at intervals remain unclear. The purpose of this study was to identify the changes in gene expression profile and to observe the role of nuclear factor kappa B (NF-kappaB) on the pathological change of renal tissue in Thy-1 N rats. Our results showed that the pathological changes of GMCs in Thy-1 N included three phases: apoptosis (40 min), necrosis (24 h) and proliferation (7 days). Concomitantly, at 40 min and on day 7, the up-regulation of 341 genes and 250 genes were observed, while 392 genes and 119 genes were down-regulated in Thy 1 N. Expression of interleukin (IL)-1beta, IL-6, proliferating cell nuclear antigen, alpha-smooth muscle actin, collagen type IV and excretion of urinary protein was increased in rats with Thy-1 N and decreased in pyrrolidine dithiocarbamate-treated rats with Thy-1 N. These data indicated that the significant changes in the gene profile were coupled with the pathological changes of Thy-1 N, and activation of NF-kappaB may contribute to the pathogenesis of GMCs apoptosis, proliferation, extracelluar matrix accumulation and proteinuria in Thy-1 N. PMID- 18422732 TI - Human immunodeficiency virus (HIV)-specific T helper responses fail to predict CD4+ T cell decline following short-course treatment at primary HIV-1 infection. AB - Early anti-retroviral treatment (ART) in primary human immunodeficiency virus (HIV) infection (PHI) may have unique, restorative immunological and virological benefits which could enhance clinical outcomes. However, the sustainability of these HIV-specific immune responses and their impact on clinical outcome remains unclear. We present a 3-year longitudinal clinical and immunological follow-up of a single-arm, prospective study assessing the long-term impact of a short-course of ART (SCART) during PHI. Twenty-eight subjects with defined PHI received 3 months of SCART at HIV-1 seroconversion. HIV-specific interferon-gamma+ CD4+ T cell responses, CD4 cell counts and plasma viral loads were assessed prospectively. Clinical outcome was defined as the time taken from PHI to a fall in CD4 cell counts <350 cells/mul on two or more occasions. Of 28 patients, 25 (89%) had detectable HIV-specific CD4+ helper responses at baseline. Five of 11 (45%) patients had preserved HIV-specific CD4+ responses 3 years after stopping SCART. Neither the presence nor magnitude of HIV-1-specific T helper responses either at baseline or 3 years following SCART cessation predicted clinical outcome. Rebound viraemia associated with stopping SCART did not diminish HIV-1 specific CD4+ responses. Long-term (>3 years) preservation of virus-specific CD4+ cells occurred in 45% of patients receiving SCART in PHI. There was no correlation between either the presence or magnitude of these responses and clinical outcome. PMID- 18422733 TI - Clonal relationship between infiltrating immunoglobulin G4 (IgG4)-positive plasma cells in lacrimal glands and circulating IgG4-positive lymphocytes in Mikulicz's disease. AB - Mikulicz's disease (MD) is gaining acceptance as an immunoglobulin G4 (IgG4) related disease characterized by bilateral lacrimal and salivary gland swelling. The aetiology of MD and other IgG4-related diseases is still unclear. The present work was performed to study the clonality of infiltrating IgG4-positive plasma cells in lacrimal glands and circulating peripheral blood cells in patients with MD, and compare the clonal relationship between infiltrating and circulating IgG4 positive cells. Total cellular RNA was extracted from the lacrimal glands and peripheral blood in five MD patients. Reverse transcription polymerase chain reaction was performed with primers specific for activation-induced cytidine deaminase (AID) and for Ig VH and IgG4. Sequences of Ig VH were compared with the structure of Ig VH of the lacrimal glands and the peripheral blood cells. AID was expressed to varying degrees in lacrimal glands of all MD patients. Most IgG4 positive cells infiltrating lacrimal glands and in peripheral blood were polyclonal, although several clonally related pairs were detected. In one patient, two of the circulating IgG4 VH4-59 clones shared identical CDR3 sequences with the clones within the lacrimal glands. In conclusion, while most tissue-infiltrating and circulating IgG4-positive cells in MD are polyclonal, some clonally related IgG4 positive cells exist between lacrimal gland and peripheral blood, accounting for the clinical features of MD as an IgG4-related disease involving multiple organs. PMID- 18422734 TI - T helper 1 (Th1)/Th2 cytokine expression shift of peripheral blood CD4+ and CD8+ T cells in patients at the post-acute phase of stroke. AB - Local humoral and cellular immune responses modulate the inflammatory processes involved in the development of atherosclerotic lesions, as well as in the evolution of brain infarcts in stroke patients. The role of systemic adaptive immunity on the progression of such disease manifestations is less clear. In the current study, we evaluated the percentages of T helper 1 (Th1) [interleukin (IL) 2, interferon (IFN)-gamma] and Th2 (IL-4, IL-10) cytokine-producing peripheral blood CD4+ and CD8+ T cells in 23 patients with a history of ischaemic stroke (IS) at the chronic stable phase of the disease (median post-stroke time 34.5 months). Seven stroke-free individuals matched for age and vascular risk factors (matched controls, MC) were collected for comparison. To measure cytokine values at baseline and after stimulation, we used a flow cytometry method of intracellular cytokine staining. Intrinsic Th1 and Th2 cytokine production in unstimulated T cells was negligible in all study participants. Following mitogenic stimulation with phorbol 12-myristate13-acetate/ionomycin, both the IS and the MC groups exhibited a similarly strong Th1 response; IL-2 production predominated in the CD4+ T cells and IFN-gamma in the CD8+ T cells. However, when measuring the Th2 cytokine-production capacity post-stimulation, a significant increase in the percentage of IL-4-producing T cells was observed in the IS groups, compared with the MC group, resulting in a significantly lower ratio of IFN-gamma-/IL-4-producing T cells. No such Th2 enhancement could be confirmed for the case of IL-10. We propose that in IS patients there is a systemic shift of the immune system towards Th2 responses at the late post-acute phase of stroke. PMID- 18422735 TI - Clinical immunology review series: an approach to the patient with recurrent superficial abscesses. AB - Patients may be referred to the immunology clinic for investigation of recurrent superficial abscess formation. In the majority of adult patients this clinical presentation does not equate with an underlying primary immune deficiency. Nevertheless, recurrent mucocutaneous abscesses can be associated with significant morbidity and long-term complications, including scarring and fistula formation, and may be associated with underlying immune-mediated disease. This review sets out an approach to the patient with recurrent superficial abscesses, focusing on the differential diagnoses, investigation and management of both the common causes and those associated with specific immune deficiency. PMID- 18422736 TI - Live probiotic Bifidobacterium lactis bacteria inhibit the toxic effects induced by wheat gliadin in epithelial cell culture. AB - Wheat gliadin induces severe intestinal symptoms and small-bowel mucosal damage in coeliac disease patients. At present, the only effective treatment for the disease is a strict life-long gluten-free diet. In this study we investigated whether probiotics Lactobacillus fermentum or Bifidobacterium lactis can inhibit the toxic effects of gliadin in intestinal cell culture conditions. The ability of live probiotics to inhibit peptic-tryptic digested gliadin-induced damage to human colon cells Caco-2 was evaluated by measuring epithelial permeability by transepithelial resistance, actin cytoskeleton arrangements by the extent of membrane ruffling and expression of tight junctional protein ZO-1. B. lactis inhibited the gliadin-induced increase dose-dependently in epithelial permeability, higher concentrations completely abolishing the gliadin-induced decrease in transepithelial resistance. The same bacterial strain also inhibited the formation of membrane ruffles in Caco-2 cells induced by gliadin administration. Furthermore, it also protected the tight junctions of Caco-2 cells against the effects of gliadin, as evinced by the pattern of ZO-1 expression. We conclude thus that live B. lactis bacteria can counteract directly the harmful effects exerted by coeliac-toxic gliadin and would clearly warrant further studies of its potential as a novel dietary supplement in the treatment of coeliac disease. PMID- 18422737 TI - Redox signalling and the inflammatory response in rheumatoid arthritis. AB - Reactive oxygen species (ROS) are produced mainly during oxidative phosphorylation and by activated phagocytic cells during oxidative burst. The excessive production of ROS can damage lipids, protein, membrane and nucleic acids. They also serve as important intracellular signalling that enhances the inflammatory response. Many studies have demonstrated a role of ROS in the pathogenesis of inflammatory chronic arthropathies, such as rheumatoid arthritis. It is known that ROS can function as a second messenger to activate nuclear factor kappa-B, which orchestrates the expression of a spectrum of genes involved in the inflammatory response. Therefore, an understanding of the complex interactions between these pathways might be useful for the development of novel therapeutic strategies for rheumatoid arthritis. PMID- 18422738 TI - Study of Toll-like receptor gene loci in sarcoidosis. AB - Sarcoidosis is a multi-factorial systemic disease of granulomatous inflammation. Current concepts of the aetiology include interactions of unknown environmental triggers with an inherited susceptibility. Toll-like receptors (TLRs) are main components of innate immunity and therefore TLR genes are candidate susceptibility genes in sarcoidosis. Ten members of the human TLR gene family have been identified and mapped to seven chromosomal segments. The aim of this study was to investigate all known TLR gene loci for genetic linkage with sarcoidosis and to follow positive signals with different methods. We analysed linkage of TLR gene loci to sarcoidosis by use of closely flanking microsatellite markers in 83 families with 180 affected siblings. We found significant linkage between sarcoidosis and markers of the TLR4 gene locus on chromosome 9q (non parametric linkage score 2.63, P = 0.0043). No linkage was found for the remaining TLR gene loci. We subsequently genotyped 1203 sarcoidosis patients from 997 families, 1084 relatives and 537 control subjects for four single nucleotide polymorphisms of TLR4, including Asp299Gly and Thr399Ile. This genotype data set was studied by case-control comparisons and transmission disequilibrium tests, but showed no significant results. In summary, TLR4 - w ith significant genetic linkage results - appears to be the most promising member of the TLR gene family for further investigation in sarcoidosis. However, our results do not confirm the TLR4 polymorphisms Asp299Gly and Thr399Ile as susceptibility markers. Our results rather point to another as yet unidentified variant within or close to TLR4 that might confer susceptibility to sarcoidosis. PMID- 18422739 TI - Exon 19 of EGFR mutation in relation to the CA-repeat polymorphism in intron 1. AB - Epidermal growth factor receptor (EGFR) mutations in lung cancer enhance tyrosine kinase activity and increase sensitivity to the EGFR tyrosine kinase inhibitor, gefitinib. Mutation analysis of the EGFR gene is therefore indispensable for predicting gefitinib response. We investigated a CA-repeat polymorphism in the EGFR gene related to EGFR mutations. Because an increasing number of CA-repeats at intron 1 of the EGFR gene has been reported to reduce transcription activity, we examined the relationship between EGFR mutations and this CA-repeat polymorphism. EGFR mutations at exon 19 were closely associated with shorter CA repeat length in the shorter allele, but this was not the case for EGFR mutations at exons 18 or 21. Increased intrinsic EGFR mRNA expression in non-cancerous lung tissues from lung adenocarcinoma patients was also significantly associated with shorter CA-repeat length. A higher frequency of EGFR mutations at exon 19 was associated with shorter CA-repeat length only in patients with high levels of EGFR mRNA expression. To determine the phenotypes of cells possessing shorter CA repeats, an in vitro study using human bronchial epithelial cells with different CA-repeat lengths was performed; more rapid cell growth and activated EGF/EGFR signaling were found more often in the cells having both shorter CA-repeats and increased EGFR mRNA expression. These results suggest that CA-repeat length in the EGFR gene may be a genetic factor related to cancer in the case of EGFR mutations at exon 19. The mechanism likely involves enhanced intrinsic expression of EGFR mRNA and activated EGF/EGFR signaling that accompany shorter CA-repeats. PMID- 18422740 TI - Plasma matrix metalloproteinase-7 as a metastatic marker and survival predictor in patients with renal cell carcinomas. AB - We evaluated the clinical usefulness of plasma matrix metalloproteinase-7 (MMP-7) as a diagnostic and prognostic biomarker in patients with renal cell carcinoma (RCC). MMP-7 was quantified in plasma of 50 healthy subjects and 97 RCC patients using a Fluorokine MultiAnalyte Profiling assay. RCC patients were stratified into the following groups: without metastases (N0M0; n = 39), with lymph nodes (N1M0; n = 13), and with distant metastases (M1; n = 45). Diagnostic performance of MMP-7 was analyzed by the receiver operating characteristics (ROC) curve. Kaplan-Meier analysis and the Cox regression model were used to estimate the impact of MMP-7 on the cancer-specific survival outcome of RCC patients. MMP-7 was significantly higher in both metastatic groups N1M0 and M1 (medians, 3.82 and 3.34 microg/L) compared to N0M0 group or controls (medians, 1.85 and 1.64 microg/L; all P < 0.001). In ROC analysis, the area under the ROC curve of MMP-7 was 0.80 in the detection of metastases in RCC (P < 0.0001). In the Kaplan-Meier analysis, patients with MMP-7 above the 95th percentile of controls showed less favorable survival rates compared to those with normal MMP-7 (log-rank test, 15.7; P < 0.0001). High MMP-7 was associated with cancer-related mortality estimated by univariate Cox regression (risk ratio, 4.34, 95% CI, 1.12-10.6; P = 0.032). The multivariate Cox regression model determined MMP-7 (risk ratio, 2.70, 95% CI, 1.39-5.24; P = 0.003) and metastases (risk ratio, 5.81, 95% CI, 2.77 12.2; P < 0.0001) as independent determinants of cancer-related survival outcomes. In conclusion, increased plasma MMP-7 could be related to metastatic disease and poor prognosis in patients with RCC. PMID- 18422741 TI - Reduced risk of endometrial cancer from alcohol drinking in Japanese. AB - The role of alcohol consumption in the etiology of endometrial cancer has not been clarified. To examine the association between alcohol consumption and endometrial cancer risk, we conducted a case-control study with 148 histologically diagnosed incident endometrial cancer cases and 1468 matched non cancer controls. Median consumption of alcohol was only 19.3 g/week among cases who drank and 28.2 g/week among controls who drank. These values are lower than in Western countries. Relative risk was analyzed in subjects classified into four groups according to weekly alcohol consumption (non-drinkers, 1-24 g/week, 25-175 g/week, and >175 g/week). Confounder-adjusted odds ratios for those consuming alcohol at <25 g/week, 25-175 g/week, and >175 g/week compared to non-drinkers were 0.79 (95% confidence interval (CI), 0.49-1.28), 0.42 (95% CI, 0.23-0.79), and 0.47 (95% CI, 0.14-1.58), respectively. Further analysis was conducted concerning self-reported physical reaction to alcohol. Among women without flushing after drinking, a significant inverse association between risk and alcohol intake was seen (trend P = 0.001). In contrast, no protective effect of alcohol was seen among women who experience flushing after drinking. These results suggest the presence of an inverse association between alcohol drinking and endometrial cancer risk among Japanese women, and that this association is evident among those without flushing. Further investigation of these findings is warranted. PMID- 18422742 TI - Overexpression of calcineurin B subunit (CnB) enhances the oncogenic potential of HEK293 cells. AB - Calcineurin (CaN) is a Ca(2+)/calmodulin (CaM)-stimulated protein phosphatase. It is a heterodimeric enzyme consisting of a catalytic A subunit (CnA) and a Ca(2+) binding regulatory B subunit (CnB). CaN's key role in vivo is well known, while the function of CnB keeps unclear except that it acts as a regulator of CaN. The present paper shows that CnB overexpression promotes proliferation of human embryonic kidney HEK293 cells by comparing with vector control cells in the complete or serum reduced medium. Furthermore, stable CnB transfectants showed dramatically improved growth in soft agar. And the migration ability of CnB overexpressors also was enhanced remarkably. But in the progress of transformation, the activity of CaN remained unchanged between CnB overexpressors and controls. Therefore, CnB, rather than CaN, is involved in the proliferation promotion of HEK293 cells. Subsequently, 11 proteins with different expression levels between CnB transfectants and controls were identified using two dimensional gel electrophoresis and electrospray ionization time-of-fight mass spectrometry. Therein, the expression of heat shock protein 27 (HSP27) and protein DJ-1 increased along with CnB overexpression. The important role of CnB in cell neoplastic transformation was found and the possible mechanism was analyzed. PMID- 18422743 TI - Role of human T-cell leukemia virus type I Tax in expression of the human telomerase reverse transcriptase (hTERT) gene in human T-cells. AB - The viral product Tax encoded by human T-cell leukemia virus type I (HTLV-I) is thought to play a central role in leukemogenesis. Clonal expansion of HTLV-I infected cells requires the extension of cell division with telomere maintenance, which is regulated by the ribonucleoprotein enzyme telomerase. However, the roles of Tax in the expression of telomerase activity in T-cells remains controversial. Our previous study indicated that expression of the human telomerase reverse transcriptase subunit (hTERT) gene, which determines telomerase activity, is tightly regulated in human T-cells. In the present study, we investigated Tax mediated regulation of hTERT gene expression by Tax in human T-cells. HTLV-I Tax induced expression of the hTERT gene in human peripheral blood leukocytes. Reporter assays revealed that Tax activated the hTERT promoter in quiescent Kit 225 cells, while the promoter activity was repressed by Tax in proliferating Jurkat cells. Both up-regulation and down-regulation by Tax were mediated through the 43-bp sequences in the promoter, which carried at least two elements that independently functioned as repressors. The two elements bound distinct factors. G1 to S phase transition induced by introduction of either cyclin D2 with cdk4 or p130-specific shRNA also activated the hTERT promoter, implying that activation of the hTERT promoter in quiescent Kit 225 cells is associated with cell cycle progression. Our findings suggest that the cell cycle state critically influences Tax-mediated regulation of hTERT expression. PMID- 18422744 TI - Epigenetic regulator polycomb group protein complexes control cell fate and cancer. AB - The chromatin-associated Polycomb group (PcG) proteins were first identified in genetic screens for homeotic transformations in Drosophila melanogaster. Besides body patterning, members of the PcG are now known to regulate epigenetic cellular memory, stem cell self-renewal, and cancer development. Here, we discuss the multifarious functions of the PcG family, isoforms of protein complexes, and its enzymatic activities, for example histone methylation, links to DNA methylation, its phosphorylation status, H2A mono-ubiquitination, SUMOylation, and links to non-coding RNA. We also discuss the function of cytosolic PcG complexes as a regulator of receptor-induced actin polymerization and proliferation in a methylation-dependent manner. We propose that the functional versatility of PcG protein complexes contributed significantly to the complexity of heritable gene repression mechanisms, signal transduction, and cell proliferation in cancer development. PMID- 18422745 TI - Midkine and its clinical significance in endometrial carcinoma. AB - Midkine (MK) is a secreted heparin-binding growth factor. Several types of human cancer have increased MK expression with elevated serum levels. The purpose of this study was to determine whether MK was expressed in endometrial carcinoma and to evaluate the clinicopathological significance of serum MK in patients with endometrial carcinoma. Immunohistochemical expression of MK was evaluated in 85 endometrial carcinoma samples and 33 controls. MK expression was significantly higher in the carcinomas than in normal endometrium (P < 0.001). Interestingly, MK expression was highest at the margins of invasion and low in the superficial areas of the tumor samples. Using ELISA, we compared serum MK concentration in 120 endometrial carcinoma patients with the concentration in 46 patients with benign gynecologic tumors. Serum MK value in patients with cancer was significantly higher than that in the patients with benign diseases (P = 0.01). Patients with positive lymph node metastasis or recurrence, or cancer death, had a higher serum MK level (P = 0.008, P = 0.009, respectively). In conclusion, MK immunoreactivity in endometrial carcinoma is significantly higher than in normal endometrium. Additionally, preoperative serum MK levels are significantly correlated with prognosis and the presence of lymph node metastasis. Thus, MK may be a useful serum biomarker for identifying high risk patients of endometrial carcinoma. PMID- 18422746 TI - Sonic hedgehog derived from human pancreatic cancer cells augments angiogenic function of endothelial progenitor cells. AB - Hedgehog signaling is important in the pathogenesis of pancreatic cancer. Several recent observations suggest the involvement of sonic hedgehog (SHH) in postnatal neovascularization. We identified a novel role for SHH in tumor-associated angiogenesis in pancreatic cancer. Immunohistochemical analysis revealed that patched homolog 1 (PTCH1), both a receptor for and transcriptional target of hedgehog signaling, was expressed in a small fraction of endothelial cells within pancreatic cancer, but not in normal pancreatic tissue. When endothelial progenitor cells (EPC) isolated from human peripheral blood were cultured with supernatant from SHH-transfected 293 cells or pancreatic cancer cells, mRNA levels of vascular endothelial growth factor (VEGF), stromal cell-derived factor 1 and angiopoietin-1 were significantly increased, whereas no such induction was observed in human umbilical vein endothelial cell (HUVEC) and human dermal microvascular endothelial cell (HMVEC). HUVEC tube formation was stimulated when cocultured with EPC, and preconditioning EPC with supernatant from KP-1 N pancreatic cancer cells highly expressing SHH significantly enhanced the effect. The effect was partially attenuated by specific inhibition of SHH with cyclopamine or a neutralizing antibody. These findings suggest that tumor-derived SHH can induce angiogenesis, and this is mediated by its effects on EPC specifically. Targeting SHH would be a novel therapeutic approach that can inhibit not only proliferation of cancer cells but also EPC-mediated angiogenesis. PMID- 18422747 TI - Anticancer activity of RecQL1 helicase siRNA in mouse xenograft models. AB - Small interfering RNAs (siRNAs) are expected to have a medical application in human therapy as drugs with a high specificity for their molecular target mRNAs. RecQL1 DNA helicase in the human RecQ helicase family participates in DNA repair and recombination pathways in the cell cycle of replication. Silencing the RecQL1 expression by RecQL1-siRNA induces mitotic death in vitro specifically in growing cancer cells. By contrast, the same RecQL1 silencing does not affect the growth of normal cells, emphasizing that RecQL1 helicase is an ideal molecular target for cancer therapy. In this study, we show that local and systemic administration of RecQL1-siRNA mixed with polyethyleneimine polymer or cationic liposomes prevented cancer cell proliferation in vivo in mouse models of cancer without noticeable adverse effects. The results indicate that RecQL1-siRNA in a complex with a cationic polymer is a very promising anticancer drug candidate, and that in particular, RecQL1-siRNA formulated with a cationic liposome has an enormous potential to be used by intravenous injection for therapy specific for liver cancers, including metastasized cancers from the colon and pancreas. PMID- 18422748 TI - Opposing effects of interferon-alpha and interferon-gamma on the expression of major histocompatibility complex class I chain-related A in tumors. AB - Natural killer cells are an important component of innate resistance to viruses, bacteria, certain parasites, and tumors. The activating receptor natural killer group 2D (NKG2D) plays a critical role in the elimination of tumor cells by cytotoxic effector cells. It has been shown that the strength of an antitumor immune response might be critically dependent on NKG2D ligard surface levels. Thus, it is essential to regulate the expression of NKG2D ligands in order to ensure effective tumor immunosurveillance and the elimination of pathogen infected cells. In the present study, we found that interferon (IFN)-alpha and IFNgamma exert opposing effects on major histocompatibility complex class I related chain A (MICA) expression in human tumor cells. IFNalpha promotes expression of the NKG2D ligand MICA in tumor cells and therefore enhances their sensitivity to natural killer lysis. In contrast, IFNgamma exerts the opposite effect. IFNalpha promotes MICA expression at the level of transcription by augmenting MICA promoter activity. IFNgamma modulates MICA expression not only at the transcriptional level, but also at the post-translational level by promoting proteolytic cleavage by matrix metalloproteinases. Further study is needed to clarify the precise regulatory mechanisms. The pathways involved in NKG2D ligand induction might represent a promising target for improving immune responses to cancer or infections. PMID- 18422749 TI - Induction of hepatocyte growth factor activator gene expression under hypoxia activates the hepatocyte growth factor/c-Met system via hypoxia inducible factor 1 in pancreatic cancer. AB - Hepatocyte growth facor activator (HGFA) is a serine protease that converts hepatocyte growth factor (HGF) into its active form. Our previous study demonstrated that tumor-stromal interaction under hypoxia augments the aggressive invasive features of pancreatic cancer line PK8 through activated HGF/c-Met signaling. The present study investigated whether or not hypoxia increases HGFA expression in PK8 cells and promotes the processing of HGF, and leads to c-Met activation. Moreover, HGFA promoter assays were performed to define whether hypoxia inducible factor-1 alpha (HIF-1alpha) directly activates the HGFA promoter in a hypoxia-dependent fashion. As a result, hypoxia induced the HGFA mRNA and protein expression in PK8 and the elevation under hypoxia was inhibited by the transfection of HIF-1alpha siRNA, thus indicating HIF-1alpha-dependent induction of HGFA. The transfection of siRNA against HGFA to PK8 cells suppressed the conversion to the active HGF, which is secreted from fibroblast MRC5. Furthermore, the phosphorylation of c-Met and cancer invasion of PK8 cells were decreased by the transfection of HGFA siRNA under hypoxia. Using the luciferase reporter system, HIF-1alpha was shown to transactivate the HGFA promoter under hypoxia. These experiments demonstrated for the first time that HGFA is a novel HIF-1 target gene. Under hypoxia, HGFA might be overexpressed and secreted from pancreatic cancer cells, which contributes to accelerate processing of HGF from fibroblast, resulting in the activation of the c-Met pathway. HGF/HGFA/c-Met recruited between cancer-stromal fibroblasts is activated under hypoxic conditions and therefore might play a central role in the aggressive invasion of pancreatic cancer. PMID- 18422750 TI - Interaction of Ganoderma triterpenes with doxorubicin and proteomic characterization of the possible molecular targets of Ganoderma triterpenes. AB - Triterpenes are the main components with cytotoxicity in Ganoderma lucidum, which is used popularly as a complementary treatment for cancer therapy in traditional Chinese medicine. To investigate the possible interaction between chemotherapeutic agents and triterpenes extracted from G. lucidum, the cytotoxicity of doxorubicin (DOX) combined with Ganoderma triterpenes (GTS) or lucidenic acid N (LCN), a purified compound, was examined in HeLa cells. The combinations targeting DOX with GTS or LCN resulted in a synergistic interaction in HeLa cells. Moreover, to identify the molecular targets of GTS, two dimensional gel electrophoresis-based comparative proteomics was carried out and proteins with altered expression levels after GTS treatment in HeLa cells were identified by matrix-assisted laser desorption/ionization time-of-flight tandem mass spectrometry. The results of our proteomic study indicated that the GTS treatment caused regulated expression of 14 proteins, which play important roles in cell proliferation, the cell cycle, apoptosis, and oxidative stress. Flow cytometric analysis confirmed that GTS could induce weak G(0)-G(1) phase arrest and combined use of GTS with DOX could induce apoptosis in cells. Furthermore, GTS enhanced the reactive oxygen species (ROS)-producing effect of DOX, and a ROS scavenger could affect the synergism between GTS and DOX. In cells with high Ku80 protein expression, the synergism between GTS and DOX was also partly affected. Importantly, in cells with high Ku80 expression that were treated with a ROS scavenger, the synergism between GTS and DOX totally disappeared. These results suggest that the synergism between GTS and DOX might be based on GTS-induced sensitization of cells to chemotherapeutics through enhanced oxidative stress, DNA damage, and apoptosis. PMID- 18422751 TI - Dendritic cells pulsed with alpha-fetoprotein and mutant P53 fused gene induce bi targeted cytotoxic T lymphocyte response against hepatic carcinoma. AB - Dendritic cell (DC)-based immunotherapy is rapidly emerging as a promising treatment in cancer therapy. We had previously shown that DC pulsed with either defined mRNA of tumor antigen (Ag) such as alpha-fetoprotein (AFP), or total RNA of hepatocellular carcinoma (HCC) could elicit Ag-specific cytotoxic T lymphocyte (CTL) response. Therefore, we suggested a novel DC-based therapeutic method, in which DCs derived from CD34(+) cells enriched peripheral blood mononuclear cells were pulsed with liposome-coated AFP and mutant P53 (mtP53) fused gene pEGFP C3/AFP-mtP53 to induce bi-targeted specific CTL responses against HCC. Three different genotype HCC cell lines, HepG2 (human histocompatibility leukocyte antigens (HLA) A2 positive, AFP expressing positive, P53 expressing negative), SMMC7721 (HLA A2 positive, neither AFP nor P53 expressing positive), and HMCC97 (HLA A2 positive, both AFP and P53 expressing positive) were selected as targets for CTL responses. An important finding was that DCs pulsed with the liposome coated fused gene could evoke more intensive bi-targeted Ag-specific CTL responses against HMCC97 than DCs pulsed with either AFP or P53 single gene (P < 0.05). This experimental therapeutic model provides a new promising cytotherapeutic approach, in that DCs pulsed with the fused gene of different Ags might induce more extensive multitargeted antitumor immunity. PMID- 18422752 TI - Individual tumorigenesis pathways of sporadic colorectal adenocarcinomas are associated with the biological behavior of tumors. AB - Clinicopathologic features of sporadic colorectal adenocarcinomas were compared using integrated data from 224 [corrected] patients subjected to curative resection. Individual steps in the tumorigenesis pathway, that is, adenomatosis polyposis coli (APC), Wnt-activated, base excision repair mutations, mismatch repair defects, RAF-mediated, transforming growth factor (TGF)-beta-suppressed, bone morphogenic protein (BMP)-suppressed, and p53 alterations, were examined in terms of genetic and epigenetic changes, as well as protein expression. Genetic and molecular alterations of right colon cancers were distinct from those of left colon and rectal cancers. Rectal cancers showed the attenuated phenotype of left colon cancers. Tumors most frequently displayed either TGF-beta- or BMP suppressed alterations (81.2%), followed by RAF-mediated alterations (78.6%), and mismatch repair defects (38.4%), constituting a total of 24 integrated pathways. Tumors lacking APC mutations or carrying the RAF alteration (V600E) were frequently associated with lymphovascular invasion and lymph node metastasis (P < 0.05). Poorly differentiated or mucinous adenocarcinomas were generally associated with high level microsatellite instability, Axin2 suppression, TGF beta1 or BMPR1A suppression, loss of heterozygosity of D18S46 or D18S474, and absence of base excision repair mutations (P < 0.0001-0.05). Early tumor recurrence was significantly correlated with lack of APC mutations (P = 0.036). Moreover, tumors that concurrently displayed APC/Wnt-activated, TGF-beta/BMP suppressed, and p53 alterations were significantly predisposed to early recurrence (P = 0.026). Our data clearly indicate that particular steps or pathways of colorectal tumorigenesis are closely associated with characteristic clinicopathologic features that, in turn, determine biological behavior, such as tumor growth, invasion, and recurrence. PMID- 18422753 TI - High LET radiation enhances apoptosis in mutated p53 cancer cells through Caspase 9 activation. AB - Although mutations in the p53 gene can lead to resistance to radiotherapy, chemotherapy and thermotherapy, high linear energy transfer (LET) radiation induces apoptosis regardless of p53 gene status in cancer cells. The aim of this study was to clarify the mechanisms involved in high LET radiation-induced apoptosis. Human gingival cancer cells (Ca9-22 cells) containing a mutated p53 (mp53) gene were irradiated with X-rays, C-ion (13-100 KeV/microm), or Fe-ion beams (200 KeV/microm). Cellular sensitivities were determined using colony forming assays. Apoptosis was detected and quantified with Hoechst 33342 staining. The activity of Caspase-3 was analyzed with Western blotting and flow cytometry. Cells irradiated with high LET radiation showed a high sensitivity with a high frequency of apoptosis induction. The relative biological effectiveness (RBE) values for the surviving fraction and apoptosis induction increased in a LET-dependent manner. Both RBE curves reached a peak at 100 KeV/microm, and then decreased at values over 100 KeV/microm. When cells were irradiated with high LET radiation, Caspase-3 was cleaved and activated, leading to poly (ADP-ribose) polymerase (PARP) cleavage. In addition, Caspase-9 inhibitor suppressed Caspase-3 activation and apoptosis induction resulting from high LET radiation to a greater extent than Caspase-8 inhibitor. These results suggest that high LET radiation enhances apoptosis by activation of Caspase-3 through Caspase-9, even in the presence of mp53. PMID- 18422754 TI - Identification of an epitope derived from CML66, a novel tumor-associated antigen expressed broadly in human leukemia, recognized by human leukocyte antigen-A*2402 restricted cytotoxic T lymphocytes. AB - CML66 is a newly identified differentiation antigen that is expressed broadly in human leukemia and solid tumors, but its physiological function remains unknown. In the present study, to clarify the feasibility of CML66-targeted cancer immunotherapy, we attempted to identify cytotoxic T lymphocyte (CTL) epitopes derived from CML66. An immunogenic CML66-derived epitope (amino acid residues 76 84; YYIDTLGRI) capable of inducing human leukocyte antigen (HLA)-A*2402 restricted CTL specific for this peptide was identified. CML66-derived peptide specific CTL efficiently lysed human leukemia cells, but not normal cells, in a HLA-A*2402-restricted fashion. Quantitative real-time polymerase chain reaction revealed that CML66 mRNA is expressed abundantly in primary acute myeloid leukemia cells, acute lymphoid leukemia cells, and chronic myelogenous leukemia cells in advanced phase, and that the expression level of CML66 mRNA in normal cells is low compared with that in leukemia cells. CML66-specific CTL precursors were detected in the peripheral blood of patients with acute leukemia. These data indicate that the CML66-derived epitope identified in the present study is a new target antigen for cellular immunotherapy of human leukemia. PMID- 18422755 TI - Interleukin-2 potentiation of cetuximab antitumor activity for epidermal growth factor receptor-overexpressing gastric cancer xenografts through antibody dependent cellular cytotoxicity. AB - Cetuximab, a chimeric monoclonal antibody to epidermal growth factor receptor (EGFR), has been proved to have clinically significant antitumor activity against advanced colorectal cancers, but its therapeutic activity for gastric cancers remains unclear. In the present study, we investigated the antitumor effect and action mechanism of cetuximab using EGFR high-expressing (MKN-28) and EGFR low expressing (GLM-1) gastric cancer cell lines without gene amplification. Cetuximab showed neither significant growth inhibition nor induction of apoptosis in either cell line in vitro, and only slightly inhibited ligand-induced phosphorylation of protein kinase B and extracellular signal-regulated kinase in MKN-28 cells. In contrast, cetuximab significantly inhibited subcutaneous and intraperitoneal tumor growth of MKN-28 cells, but not GLM-1 cells, in nude mice. This antitumor activity was significantly enhanced and diminished in nude mice by treatment with interleukin-2 (IL-2) and antiasialo GM1 antibody, which can expand and deplete natural killer (NK) cells, respectively. Antibody-dependent cellular cytotoxicity (ADCC) of cetuximab, as measured by (51)Cr release assay, was significantly higher in MKN-28 than in GLM-1 cells. This ADCC activity was enhanced by IL-2 and reduced by heat-aggregate of human immunoglobulin G, an inhibitor for FcR-III of NK cells. These results suggest that cetuximab in combination with IL-2 shows significant antitumor activity against EGFR high expressing gastric cancer mainly through NK cell-mediated ADCC. Combination therapy with cetuximab and IL-2 would thus offer a new potential therapeutic approach for a subset of EGFR-overexpressing gastric cancers. PMID- 18422756 TI - Coordinate suppression of Sdpr and Fhl1 expression in tumors of the breast, kidney, and prostate. AB - The Src tyrosine kinase associates with the focal adhesion adaptor protein Cas (Crk-associated substrate) to suppress the expression of potential tumor suppressor genes. For example, Src utilizes Cas to suppress the expression of the LIM-only protein Fhl1 (four and a half LIM domains 1), in order to promote non anchored tumor-cell growth and migration. Here, we report that the promoter region of the Fhl1 gene was methylated more in Src-transformed cells than non transformed cells. In addition, global expression analysis indicates that Fhl1 induced expression of serum deprivation response factor (Sdpr) in Src-transformed cells. Moreover, Fhl1 and Sdpr was expressed in approximately 87% and 40% of samples obtained from non-transformed breast, 100% of samples obtained from non transformed kidney, and over 60% of samples obtained from non-transformed prostate. In contrast, Fhl1 and Sdpr was detected in approximately 40% and 7% of matched samples from mammary carcinoma, less than 11% of matched samples from kidney carcinoma, and in less than 22% of matched samples from prostate carcinoma. These data indicate that Fhl1 and Sdpr expression was significantly reduced in tumors of the breast (P < 0.02 and P < 0.001), kidney (P < 0.01), and prostate (P < 0.05). In addition, although Src can activate mitogen-activated protein kinase (MAPK) to promote tumor-cell growth, our data indicate that Src did not rely on MAPK activity to suppress the expression of Fhl1 and Sdpr in transformed cells. Thus, Src induced methylation of the promoter region of the Fhl1 gene; Src suppressed Fhl1 and Sdpr expression independent of mitogen activated protein kinase (MAPK) activity; Fhl1 induced the expression of Sdpr in Src-transformed cells; and Fhl1 and Sdpr expression was suppressed in tumors of the breast, kidney, and prostate. PMID- 18422757 TI - Adult onset leukodystrophy with neuroaxonal spheroids: clinical, neuroimaging and neuropathologic observations. AB - Pigmented orthochromatic leukodystrophy and hereditary diffuse leukoencephalopathy with spheroids are two adult onset leukodystrophies with neuroaxonal spheroids presenting with prominent neurobehavioral, cognitive and motor symptoms. These are familial or sporadic disorders characterized by cerebral white matter degeneration including myelin and axonal loss, gliosis, macrophages and axonal spheroids. We report clinical, neuroimaging and pathological correlations of four women ages 34-50 years with adult onset leukodystrophy. Their disease course ranged from 1.5-8 years. Three patients had progressive cognitive and behavioral changes; however, one had acute onset. Neuroimaging revealed white matter abnormalities characterized by symmetric, bilateral, T2 hyperintense and T1 hypointense Magnetic Resonance Imaging signal involving frontal lobe white matter in all patients. Extensive laboratory investigations were negative apart from abnormalities in some mitochondrial enzymes and immunologic parameters. Autopsies demonstrated severe leukodystrophy with myelin and axonal loss, axonal spheroids and macrophages with early and severe frontal white matter involvement. The extent and degree of changes outside the frontal lobe appeared to correlate with disease duration. The prominent neurobehavioral deficits and frontal white matter disease provide clinical pathologic support for association pathways linking distributed neural circuits sub-serving cognition. These observations lend further support to the notion that white matter disease alone can account for dementia. PMID- 18422758 TI - Melatonin attenuates hypoxia-induced ultrastructural changes and increased vascular permeability in the developing hippocampus. AB - Hypoxic injury in the perinatal period may be involved in damaging the developing hippocampus. The damage may be mediated by excess production of vascular endothelial growth factor (VEGF) and nitric oxide (NO). We examined the hippocampus of neonatal Wistar rats subjected to hypoxia for VEGF and NO production. The mRNA and protein expression of hypoxia inducible factor-1alpha, endothelial, neuronal, inducible nitric oxide synthase and VEGF was found to be up-regulated significantly after the hypoxic exposure. Tissue VEGF concentration and NO production were also increased. By electron microscopy, swollen dendrites, vacuolated axons and hypertrophic astrocyte end feet associated with blood vessels were observed in hypoxic animals. In hypoxic rats, the passage of rhodamine isothiocyanate (RhIC) and horseradish peroxidase, administered intraperitoneally or intravenously, was observed through vascular walls. Furthermore, immunoglobulin G was localized in the neuropil and neurons. We suggest that increased VEGF and NO production in hypoxia had resulted in increased vascular permeability, leading to structural alteration of the dendrites and axons. Melatonin administration reduced VEGF and NO levels as well as leakage of RhIC, suggesting that it has a therapeutic potential in reducing hypoxia-associated damage in the developing hippocampus. PMID- 18422759 TI - Antagonism of the chemokine receptors CXCR3 and CXCR4 reduces the pathology of experimental autoimmune encephalomyelitis. AB - Chemokines regulate lymphocyte trafficking under physiologic and pathologic conditions. In this study, we have investigated the role of CXCR3 and CXCR4 in the activation of T lymphocytes and their migration to the central nervous system (CNS) using novel mutant chemokines to antagonize CXCR3 and CXCR4 specifically. A series of truncation mutants of CXCL11, which has the highest affinity for CXCR3, were synthesized, and an antagonist, CXCL11((4-79)), was obtained. CXCL11((4-79)) strongly inhibited the migration of activated mouse T cells in response to all three high-affinity CXCR3 ligands, CXCL9, 10 and 11. CXCL12((P2G2)), while exhibiting minimal agonistic activity, potently inhibited the migration of activated mouse T cells in response to CXCL12. Interfering with the action of CXCR3 and CXCR4 with these synthetic receptor antagonists inhibited experimental autoimmune encephalomyelitis (EAE), a mouse model of multiple sclerosis and reduced the accumulation of CD4(+) T cells in the CNS. Further investigation demonstrated that CXCL12((P2G2)) inhibited the sensitization phase, whereas CXCL11((4-79)) inhibited the effector phase of the immune response. Our data suggest that simultaneous targeting of CXCR4 and CXCR3 may be of benefit in the treatment of the CNS autoimmune disease. PMID- 18422760 TI - Heparan sulfate accumulation with Abeta deposits in Alzheimer's disease and Tg2576 mice is contributed by glial cells. AB - Amyloid beta-peptide (Abeta) plaques, one of the major neuropathological lesions in Alzheimer's disease (AD), can be broadly subdivided into two morphological categories: neuritic and diffuse. Heparan sulfate (HS) and HS proteoglycans (HSPGs) are codeposits of multiple amyloidoses, including AD. Although HS has been considered a limiting factor in the initiation of amyloid deposition, the pathological implications of HS in Abeta deposits of AD remain unclear. In this study, immunohistochemistry combined with fluorescence and confocal microscopy was employed to gain deeper insight into the accumulation of HS with Abeta plaques in sporadic and familial AD. Here we demonstrate that HS preferentially accumulated around the Abeta40 dense cores of neuritic plaques, but was largely absent from diffuse Abeta42 plaques, suggesting that Abeta42 deposition may occur independently of HS. A codeposition pattern of HS with Abeta deposits in Tg2576 mice was also examined. We identified the membrane-bound HSPGs, glypican-1 (GPC1) and syndecan-3 (SDC3), in glial cells associated with Abeta deposits, proximal to sites of HS accumulation. In mouse primary glial cultures, we observed increased levels of GPC1 and SDC3 following Abeta stimulation. These results suggest that HS codeposits with Abeta40 in neuritic plaques and is mainly derived from glial cells. PMID- 18422762 TI - Immunohistochemical analysis supports a role for INI1/SMARCB1 in hereditary forms of schwannomas, but not in solitary, sporadic schwannomas. AB - The INI1/SMARCB1 protein product (INI1), a component of a transcription complex, was recently implicated in the pathogenesis of schwannomas in two members of a single family with familial schwannomatosis. Tumors were found to have both constitutional and somatic mutations of the SMARCB1 gene and showed a mosaic pattern of loss of INI1 expression by immunohistochemistry, suggesting a tumor composition of mixed null and haploinsufficient cells. To determine if this finding could be extended to all tumors arising in familial schwannomatosis, and how it compares with other multiple schwannoma syndromes [sporadic schwannomatosis and neurofibromatosis 2 (NF2)] as well as to sporadic, solitary schwannomas, we performed an immunohistochemistry analysis on 45 schwannomas from patients with multiple schwannoma syndromes and on 38 solitary, sporadic schwannomas from non-syndromic patients. A mosaic pattern of INI1 expression was seen in 93% of tumors from familial schwannomatosis patients, 55% of tumors from sporadic schwannomatosis, 83% of NF2-associated tumors and only 5% of solitary, sporadic schwannomas. These results confirm a role for INI1/SMARCB1 in multiple schwannoma syndromes and suggest that a different pathway of tumorigenesis occurs in solitary, sporadic tumors. PMID- 18422761 TI - Tumor necrosis factor alpha is reparative via TNFR2 [corrected] in the hippocampus and via TNFR1 [corrected] in the striatum after virus-induced encephalitis. AB - Differentiating between injurious and reparative factors facilitates appropriate therapeutic intervention. We evaluated the role of tumor necrosis factor alpha (TNFalpha) in parenchymal brain pathology resolution following virus-induced encephalitis from a picornavirus, Theiler's murine encephalomyelitis virus (TMEV). We infected the following animals with TMEV for 7 to 270 days: B6/129 TNF(-/-) mice (without TNFalpha expression), B6/129 TNFR1(-/-) mice (without TNFalpha receptor 1 expression), and B6/129 TNFR2(-/-) mice (without TNFalpha receptor 2 expression). Normal TNFalpha-expressing controls were TMEV-infected B6, 129/J, B6/129F1 and B6/129F2 mice. Whereas all strains developed inflammation and neuronal injury in the hippocampus and striatum 7 to 21 days postinfection (dpi), the control mice resolved the pathology by 45 to 90 dpi. However, parenchymal hippocampal and striatal injury persisted in B6/129 TNF(-/-) mice following infection. Treating virus-infected mice with active recombinant mouse TNFalpha resulted in less hippocampal and striatal pathology, whereas TNFalpha neutralizing treatment worsened pathology. T1 "black holes" appeared on MRI during early infection in the hippocampus and striatum in all mice but persisted only in TNF(-/-) mice. TNFR2 [corrected] mediated hippocampal pathology resolution whereas TNFR1 [corrected] mediated striatal healing. These findings indicate the role of TNFalpha in resolution of sublethal hippocampal and striatal injury. PMID- 18422763 TI - Ocular side-effects of urological pharmacy. PMID- 18422764 TI - Long-term efficacy of duloxetine in women with stress urinary incontinence. AB - OBJECTIVE: To assess the maintenance of efficacy of duloxetine beyond 3 months, using data from several long-term, open-label studies, as the efficacy of duloxetine 40-mg twice daily for treating women with stress urinary incontinence (SUI) for up to 3 months has been established in several randomized, placebo controlled clinical trials. PATIENTS AND METHODS: Data from 1424 patients (Cohort A) enrolled in three 12-week, placebo-controlled clinical trials and their uncontrolled, open-label extensions, and in one uncontrolled, open-label study, were used to assess long-term continuation rates and continued efficacy based on responses to the validated Patient Global Impression of Improvement (PGI-I) scale for up to 30 months. Data from another 2758 patients (Cohort B) enrolled in an additional placebo-controlled study and its open-label extension were used to assess PGI-I ratings, reductions in incontinence episode frequency (IEF) recorded on urinary diaries, and the relationship between PGI-I ratings and reductions in IEF for up to 72 weeks. RESULTS: In Cohort A, the duloxetine continuation rate at 1 year was 42.5%. At 12, 24 and 30 months, most (83%, 83% and 88%, respectively) patients in Cohort A who continued treatment rated their incontinence in one of the three 'better since starting treatment' PGI-I categories. Both the median IEF reductions (50-77%) and the PGI-I 'better' ratings (70-88% of patients) remained fairly consistent over 72 weeks in Cohort B. Finally, IEF reductions increased with increasing PGI-I ratings (approximately 46% for 'a little better', 75% for 'much better' and 95% for 'very much better') over the first year of treatment. CONCLUSION: The benefits of duloxetine were maintained in patients who continued treatment for up to 30 months. However, these favourable results need to be interpreted cautiously, as many patients discontinued treatment and those with better responses are more likely to continue taking medication. PMID- 18422765 TI - Laparoscopic transposition of lower-pole crossing vessels in the management of pelvi-ureteric junction obstruction. PMID- 18422766 TI - Evaluation of cryoprobe deployment precision with body surface and in situ templates. AB - OBJECTIVE: To evaluate the precision of cryoprobe targeting with a surface template, an in situ template (on the target organ), or a combined approach. MATERIALS AND METHODS: Fourteen participants placed five 17 G cryoprobes into porcine kidneys in a laparoscopic trainer using a surface template (group 1), an in situ template (group 2) or a combination of the two templates (group 3). The distance from the ideal probe placement was measured both on the anterior and posterior aspect of the kidney. The sequence of attempts was randomized. The distances were compared across the three groups using anova with the adjustment for multiple comparisons. RESULTS: The mean distance from the ideal probe placement was 1.58 cm (anterior) and 1.81 cm (posterior) in group 1, 0.05 cm and 0.39 cm in group 2, and 0.07 cm and 0.22 cm in group 3, respectively. The placement of the probes was significantly more accurate in groups 2 (P < 0.001 anteriorly and P < 0.002 posteriorly) and 3 (P = 0.001 anteriorly and P < 0.001 posteriorly) compared with group 1. There was no significant difference between groups 2 and 3. CONCLUSION: In this in vitro model, the use of internal or combined internal and external templates allows for significantly more precise deployment of 17-G cryoprobes than a standard external template alone. PMID- 18422767 TI - The discovery and application of gene fusions in prostate cancer. AB - Chromosomal rearrangements play a causal role in haematological and mesenchymal malignancies. Importantly, the resulting gene fusions can serve as specific therapeutic targets, as exemplified by the development of imatinib (Gleevec), which specifically inhibits the BCR-ABL gene fusion product that defines chronic myeloid leukaemia. Recently, gene fusions involving the prostate-specific gene transmembrane protease, serine 2 (TMPRSS2) and members of the erythroblastosis virus E26 transforming sequence (ETS) family of transcription factors were identified in most of PSA-screened prostate cancers. In this review, we summarize the identification, characterization and detection of TMPRSS2:ETS gene fusions and their role in prostate cancer development. We also discuss the discovery of additional 5' partners that define distinct classes of ETS gene fusions based on the prostate specificity and androgen responsiveness of the 5' partner. Additionally, we also summarize conflicting reports about associations between gene fusion status and patient outcome. The specificity of ETS gene fusions in prostate cancer suggests that they may have causal roles in prostate cancer and suggest utility in prostate cancer detection, stratification and treatment. PMID- 18422768 TI - Performance and functional outcome of endoscopic extraperitoneal radical prostatectomy in relation to obesity: an assessment of 500 patients. AB - OBJECTIVE: To investigate the impact of obesity on the performance and functional outcome of endoscopic extraperitoneal radical prostatectomy (EERPE). PATIENTS AND METHODS: We retrospectively examined 500 patients treated with EERPE; they were categorized into three groups according to the World Health Organization classification of obesity: normal weight (body mass index, BMI, <25.0 kg/m(2)), overweight (25.0-29.9 kg/m(2)) and obese (30.0 kg/m(2)). The database of our institution was reviewed and perioperative data evaluated. The functional data were collected through questionnaires before and after EERPE and analysed statistically. RESULTS: The age, prostate size and preoperative PSA level were similar in all three groups. The mean (sd) BMI was 27 (3.3) kg/m(2), with 26.8%, 56.6% and 16.6% of the patients classed as normal, overweight and obese, respectively. A pelvic lymph node dissection and nerve-sparing was done in 218 and 123 patients, respectively. There was no statistically significance difference in the number of patients in each group who had previous procedures. Obese patients had a significantly higher American Society of Anesthesiologists score. The mean operative duration for all patients was 149 min; there was a statistically significant difference in duration among the three groups, with EERPE or nerve-sparing EERPE requiring a mean of 20 min more in obese patients. There was no conversion to open surgery. The estimated mean blood loss was 200 mL; four patients, none of them in the obese group, received a blood transfusion. At 3 months after EERPE there was a trend to worse continence in obese patients, but it was not statistically significant, and was not apparent at 6 months. There was no difference in transfusion rate and duration of catheterization. CONCLUSION: EERPE seems to be a feasible and reproducible surgical technique in obese patients, although the operation takes longer. PMID- 18422769 TI - 'Crushing the stone': a brief history of lithotripsy, the first minimally invasive surgery. AB - OBJECTIVE: To trace the history of lithotripsy, marking the start of minimally invasive surgery. METHODS: Primary and secondary sources describing the development of lithotripsy as a method to treat common bladder stones were reviewed. RESULTS: Lithotripsy emerged in the early 19th century as an alternative to the morbid and frequently fatal perineal lithotomy practised for thousands of years to relieve sufferers of bladder stones. Lithotripsy developed in stages, first by learning how to pass a straight hollow sound, then drilling stones to break them up, later by crushing them, followed by means to evacuate the fragments, finally by integrating the lithotrite with the cystoscope, permitting surgery under direct vision. CONCLUSIONS: Lithotripsy evolved as the first minimally invasive surgical procedure owing to the ingenuity and skill of our urological forebears. PMID- 18422770 TI - Differential expression of prognostic markers in histological subtypes of papillary renal cell carcinoma. AB - OBJECTIVE: To assess the expression of the tumour markers stromelysin 3, MUC1, p53 and cytokeratin-7 in papillary renal cell carcinoma (pRCC, for which two histological subtypes are distinguished, i.e. type 1 and type 2, the latter appearing to be associated with a poorer prognosis) and to determine whether any of these markers might be of prognostic value. PATIENTS AND METHODS: In a retrospective study of 50 patients, the type and nuclear grade of tumours was determined by histological analyses, the presence of microvascular emboli detected, and the markers assessed by immunohistochemical analysis using anti stromelysin 3, anti-MUC1, anti-p53 and anti-cytokeratin-7 antibodies. RESULTS: Twenty-five patients each had a type 1 or type 2 tumour. MUC1 and cytokeratin-7 were principally expressed in type 1 tumours, being detected in 76% and 84%, respectively. By contrast, p53 accumulated principally in type 2 tumours (36%); the accumulation of p53 was also associated with poorer survival. In patients with type 2 tumours with a more unfavourable development, stromelysin-3 expression was associated with a more advanced stage and a higher risk of metastases. CONCLUSION: Subtyping pRCC according to the recommended morphological criteria appears to be worthwhile, and can be reinforced by immunohistochemical tests capable of detecting cytokeratin-7 and MUC1 expression. Immunohistochemical detection of p53 is of prognostic value, as accumulation of this factor is associated with poorer survival. PMID- 18422771 TI - Early hormonal data from a multicentre phase II trial using transdermal oestrogen patches as first-line hormonal therapy in patients with locally advanced or metastatic prostate cancer. AB - OBJECTIVE: To assess the hormonal effects of Fem7 (Merck, KGaA, Darmstadt, Germany) 100 microg transdermal oestrogen patches on men undergoing first-line androgen-deprivation therapy for prostate cancer. PATIENTS AND METHODS: PATCH is a multicentre, randomized, phase II trial for men with locally advanced or metastatic prostate cancer, comparing luteinizing hormone-releasing hormone agonist therapy with oestrogen patches. To assess the dosing schedule for the patches, as this was the first time that this brand of patch had been used in men, and to reassure patients and participating clinicians, the Independent Data Monitoring Committee agreed to early release of hormonal data from this study. RESULTS: Oestradiol, testosterone and prostate-specific antigen (PSA) levels are presented for the first group of 14 patients who received the patches (with 1 withdrawal) and for whom there were > or =12 weeks of follow-up by March 2007. After 12 weeks, testosterone levels (nmol/L) in eight of the 13 patients were <1.7, two were 1.7-2 and three were >2. The median (range) serum oestradiol levels was 442 (52.1-1542) pmol/L and all patients had a PSA response, with eight having a PSA level of <4 ng/mL. CONCLUSION: These results confirm that oestrogen patches produce castrate levels of testosterone and concomitant PSA responses. They also highlighted the potential differences between different brands of oestrogen patches, and the need to monitor hormonal response, toxicity and efficacy until more experience with oestrogen patches for this clinical indication is obtained. The number of patches recommended in the PATCH study has now been increased. PMID- 18422772 TI - Vaporization of prostates of > or =80 mL using a potassium-titanyl-phosphate laser: midterm-results and comparison with prostates of <80 mL. AB - OBJECTIVE: To compare the safety and outcome of potassium-titanyl-phosphate (KTP) Greenlight (Laserscope, AMS, Minnetonka, MN, USA) vaporization for treating benign prostatic hyperplasia (BPH) in prostates of > or =80 vs <80 mL. PATIENTS AND METHODS: In all, 204 consecutive patients were enrolled into this prospective study; 31 were excluded from analysis for various reasons, thus 173 (median age 66.8 years; 39 with prostates of > or =80 mL) were evaluated for maximum urinary flow rate (Q(max)), postvoid residual urine (PVR), the International Prostate Symptom Score (IPSS), and quality-of-life (QoL) score. The median follow-up was 11.7 months. Of the 173 men, 26.6% were anaesthesiological high-risk patients and 32.4% took anticoagulants. RESULTS: The Q(max) improved in prostates of > or =80 mL from 6.9 mL/s before to 15.3 mL/s immediately after catheter removal, and 23.4 mL/s after 3 months (P < 0.001). The improvement was similar to that in smaller prostates. There were equally effective changes in PVR (P < 0.001). The IPSS decreased from 19.0 before to 7.0, 4.0 and 5.5 at 3, 6 and 12 months after surgery (P < 0.001). Changes did not differ from those in smaller glands. The results were similar for QoL (P < 0.001). During surgery there was no major bleeding and no transurethral resection syndrome. There was acute urinary retention after catheter removal in 10.4% of men; 4.6% developed urinary tract infections, whereas the rate was higher in men with larger prostates (10.3%). The re-operation rate was higher in men with larger prostates, at 23.1% vs 10.4% (P = 0.09). CONCLUSION KTP laser vaporization is a safe and effective procedure for surgically treating BPH. The functional outcome in larger prostates is similar to that in smaller glands, but there was a serious trend to a higher re-operation rate in men with larger prostates. PMID- 18422773 TI - Comparison of stage migration patterns between Europe and the USA: an analysis of 11 350 men treated with radical prostatectomy for prostate cancer. AB - OBJECTIVE: To examine the stage migration patterns in patients treated with radical prostatectomy (RP) for prostate cancer in Europe and in the USA in the last 20 years. PATIENTS AND METHODS: Between 1988 and 2005, RP was performed in 11 350 men: 5739 from Europe and 5611 from the USA. Independent-samples t-test and the chi-square test were, respectively, used for comparisons of means and proportions. The trend test was used to test the statistical significance of trends in proportions over time. RESULTS; Temporal patterns in patients' age, stage and PSA level at presentation were similar on both continents. Conversely, temporal patterns in Gleason sum distribution differed. In the USA, the rate of biopsy Gleason sums of 2-5 decreased from 32.8% to 0.2% (P < 0.001), while the rate of Gleason sums of 7 and 8-10 increased (P < 0.001). Conversely, in Europe the rate of Gleason sums of 6 increased from 40% to 64% (P < 0.001) at the expense of all other Gleason sums. At RP, the rate of Gleason sums of 2-5 decreased on both continents and the rate of a Gleason sum of 7 increased in the USA. Moreover, no important differences in pathological stage trends (organ confinement, extracapsular extension and seminal vesicle invasion) distinguished either population. Finally, the rate of lymph node involvement increased in the USA but remained stable in Europe. CONCLUSIONS: Stage and grade migration affected the USA and Europe to different extents. These differences should be accounted for when prediction tools or comparisons between the USA and Europe are considered. PMID- 18422774 TI - Watchful waiting and active surveillance: the current position. PMID- 18422775 TI - Sporadic bloody diarrhoea-associated thrombotic thrombocytopenic purpura haemolytic uraemic syndrome: an adult and paediatric comparison. AB - Although diarrhoea-associated haemolytic uremic syndrome (HUS) in children is well described, the clinical features of bloody diarrhoea-associated thrombotic thrombocytopenic purpura (TTP)-HUS in adults are not documented. Twenty-one adults, 6.5% of the 322 adults in The Oklahoma TTP-HUS Registry, 1989-2006, have presented with bloody diarrhoea. There were no case clusters. Escherichia coli O157:H7 was identified in five patients, but many patients did not have appropriate studies. The annual incidence was 0.68/10(6), 10-fold less than the incidence of diarrhoea-associated HUS in children in Oklahoma. Two (13%) of 16 patients in whom ADAMTS13 (A Disintegrin And Metalloproteinase with a ThromboSpondin type 1 motif, member 13) was measured had <10% activity. Severe neurological abnormalities (67%) and renal failure (62%) were common; seven patients (33%) died; no survivors have relapsed. Compared to the 38 other Oklahoma Registry patients with ADAMTS13 <10%, frequency of severe neurological abnormalities and death was not different; frequency of renal failure was greater; frequency of relapse was less. Compared to 5999 children with sporadic diarrhoea-associated HUS in published reports, frequency of renal failure and relapse was not different; frequency of severe neurological abnormalities and death was greater (P < 0.05 for all differences). Awareness of the continuous occurrence of sporadic bloody diarrhoea-associated TTP-HUS in adults is important for prompt diagnosis and appropriate management. PMID- 18422776 TI - Hypoxia inducible factor-alpha activation in lymphoma and relationship to the thioredoxin family. AB - Hypoxia inducible factors (HIFs) activate oncogenic pathways, while thioredoxins (Trx), including Trx1 and Trx reductases-1 and -2 (TrxR1 and TrxR2), promote HIF alpha stabilization. In immunoblotting studies in lymphoma cell lines we found that Raji and SUDHL4 cells exhibited normoxic HIF-2alpha protein stabilization. Five cell lines showed increased TrxR1 expression, while only Namalwa, HF1 and SUDHL4 had Trx1 and TrxR2 activation. Tissue microarrays in diffuse large B-cell lymphoma (DLBCL) and follicular lymphoma (FL) identified different HIF expression among histological subgroups (e.g. 44% DLBCL vs. 11% of FL cases with moderate-to high expression of HIF-1alpha and HIF-2alpha, P = 0.0017). These data demonstrate that HIF and the thioredoxin family are abnormally activated in lymphoma. PMID- 18422777 TI - Induction of gamma-globin gene transcription by hydroxycarbamide in primary erythroid cell cultures from Lepore patients. AB - Increased expression of fetal haemoglobin (HbF) may ameliorate the clinical course of beta-thalassemia and sickle cell disease. Some pharmacological agents, such as hydroxycarbamide (HC), can increase fetal haemoglobin synthesis during adult life. Cellular selection and/or molecular mechanisms have been proposed to account for this increase. To explore the mechanism of action of HC we focused on homozygous Hb-Lepore patients that presented with high fetal haemoglobin levels and were good responders to HC treatment "in vivo". We performed primary erythroid cultures from peripheral blood of four homozygous Lepore patients. The increase in HBG (gamma-globin) transcription levels and HbF content in these cultures, after HC treatment, were detected by quantitative real time polymerase chain reaction analysis and flow cytometric analysis. Primary transcript "in situ" hybridization analysis showed a 2-fold increase in the number of cells expressing both HBG alleles in HC-treated erythroid cultures. These studies, demonstrating the larger number of biallelic HBG expressing cells, suggest that HC is able to stimulate the activation of HBG transcription. These observations provide evidences that the molecular mechanism of action is involved in the increase of fetal haemoglobin production by HC. PMID- 18422778 TI - Detection of different clonal EBV strains in Hodgkin lymphoma and nasopharyngeal carcinoma tissues from the same patient. AB - The ubiquitous herpesvirus Epstein-Barr virus (EBV) is linked to the development of several malignancies, including nasopharyngeal carcinoma (UCNT) and Hodgkin lymphoma (HL). Despite the well-known oncogenic properties of the EBV latent membrane protein 1 (LMP-1), the different oncogenic pathways involved in the pathogenesis of each disease remain unclear. This study reported, for the first time, the case of a patient with sequential development of UCNT and HL. Polymerase chain reaction was used to determine the LMP-1 gene sequence and demonstrate that the two tumours contained different clonal viral genomes, suggesting a central and specific role of EBV infection. PMID- 18422780 TI - Reactivity profile of anti-factor VIII antibodies with designed synthetic peptides mimicking epitopes of the C2 and a1 domains. AB - Antibodies (Abs) that block factor VIII (FVIII) activity occur in hemophilia A patients treated with FVIII replacement therapy and severely impair treatment. In this work, we designed and synthesized ten peptides whose sequences are found in putative epitopes at the surface of a1 and C2 domains of the FVIII molecule. These peptides were screened for their ability to inhibit the binding of anti FVIII Abs from plasmas of hemophilia A patients to FVIII. All peptides were efficient in inhibiting anti-FVIII Abs in plasma from patients with inhibitors, with however different efficiencies. It was found that each tested patient's plasma had a different profile of reactivity with peptides, consistent with an individual anti-FVIII Ab specificity. The profile of recognized peptides was also changing during the treatment of the patients. Three peptides were used in an affinity chromatography assay to attempt to remove anti-FVIII Abs from patients' plasma. Anti-FVIII IgGs were significantly captured by the peptide-Sepharose affinity matrixes as assessed by enzyme-linked immunosorbent assay. However, due to the low level of Abs in the plasma samples, other methods (Chromogenic and Bethesda assays) were not sensitive enough to properly detect the reduction of inhibitors. PMID- 18422779 TI - Rapid and sustained clearance of circulating lymphoma cells after chemotherapy plus rituximab: clinical significance of quantitative t(14;18) PCR monitoring in advanced stage follicular lymphoma patients. AB - This study of first-line treatment in advanced-stage follicular lymphoma patients analysed the effects of MCP (mitoxantrone, chlorambucil and prednisolone) chemotherapy alone or in combination with rituximab (R-MCP) on circulating lymphoma cells (CLC) and assessed the prognostic value of a quantitative monitoring of CLC. CLC numbers were determined by quantitative polymerase chain reaction (PCR) for the t(14;18)-translocation or by allele-specific PCR for rearranged immunoglobulin heavy chain genes. We analysed blood samples from 43 patients treated in a randomized trial comparing eight cycles of MCP versus R MCP. Clearance of CLC at the end of therapy was achieved in 21/25 patients (84%) treated with R-MCP compared with 0/18 after MCP alone (P < 0.0001). A > or = 2 log CLC reduction was associated with a favourable clinical response (P = 0.0004) and prolonged event-free survival (P = 0.02). In R-MCP patients, stable CLC numbers or consistently PCR-negative blood samples were associated with a continuing clinical remission whereas in two patients a relapse was preceded by a > or = 2 log CLC increase. This study demonstrated that R-MCP led to a rapid and sustained eradication of CLC and a > or = 2 log CLC reduction was associated with a superior quality and duration of the clinical response. PMID- 18422781 TI - Results of bone marrow examination in 275 patients with histological features that suggest an indolent type of cutaneous B-cell lymphoma. AB - Whether or not bone marrow biopsies should be performed routinely in patients with skin lesions that show histological features consistent with an indolent B cell lymphoma [marginal zone lymphoma (MZL) or follicle centre lymphoma (FCL)] has been debated. As no studies have addressed this question for this group of lymphomas, we evaluated the results of bone marrow biopsy examination in 275 patients with histological features consistent with MZL (n = 82) or FCL (n = 193) first presenting in the skin. In the MZL group, two of 82 patients (2%) showed bone marrow involvement, which was the only extracutaneous localization in one of these patients. In the FCL group, 22 of 193 patients (11%) had bone marrow involvement and was the only extracutaneous localization in nine of them. FCL patients with skin lesions and a positive bone marrow had a significantly worse prognosis when compared with patients with only skin lesions (5-year disease specific survival 63% vs. 95%; P = 0.001). These results indicate that bone marrow investigation is essential for staging patients with a FCL first presenting in the skin. Bone marrow examination appears to have limited value in patients with MZL presenting in the skin. PMID- 18422782 TI - A new prognosticator for post-transplant lymphoproliferative disorders after renal transplantation. PMID- 18422783 TI - Developing idiotype vaccines for lymphoma: from preclinical studies to phase III clinical trials. AB - Therapeutic vaccines for B-cell non-Hodgkin lymphoma (NHL) using the clonal tumour immunoglobulin idiotype (Id) have been under development for more than three decades. A major obstacle for rapid progress in the field has been that the Id vaccine is patient-specific and required the generation of a custom-made product. The manufacturing issues were recently overcome by advances in hybridoma and recombinant DNA technology which facilitated the completion of several phase I and II clinical trials. The strong immunogenicity and apparent clinical benefit observed on the early phase studies led to the initiation of three randomized phase III clinical trials that are also nearing completion. This review will focus on the development of Id vaccines before and after the introduction of rituximab for the treatment of B-cell NHL and also discuss potential strategies to enhance the efficacy of active immunotherapy in the future. PMID- 18422785 TI - csf1 is required for early embryonic macrophage development: characterization of the csf1(op)/csf1(op) mutation in ES cell-derived macrophages. PMID- 18422784 TI - Functional analysis of single amino-acid mutations in the thrombopoietin-receptor Mpl underlying congenital amegakaryocytic thrombocytopenia. AB - Congenital amegakaryocytic thrombocytopenia (CAMT) is a rare disorder that presents with severe thrombocytopenia and absence of megakaryocytes in the bone marrow. The disease may develop into bone marrow aplasia. Genetic defects in the gene encoding the thrombopoietin (Tpo) receptor, MPL, are the cause of this disease. In a previous study, we identified four missense mutations in CAMT patients, predicting Arg102Pro, Pro136His, Arg257Cys and Pro635Leu. To investigate whether these mutations result in defective Tpo-binding and/or signalling, full-length wildtype and mutant MPL were transduced into K562 cells. Expression levels and the ability to activate the mitogen-activated protein kinase, Janus kinase-signal transducer and activator of transcription and phosphoinositide-3 kinase pathways upon Tpo-binding were studied. The results predicted that MPL carrying the P136H or P635L mutation was not properly expressed, whereas the R102P and R257C mutations resulted in impaired signal transduction. Our results indicate that a severe clinical course may be expected when these mutations lead to absent Mpl expression or signalling in CAMT patients with missense mutations. PMID- 18422786 TI - Recent advances in the pathogenesis and management of juvenile myelomonocytic leukaemia. AB - Juvenile myelomonocytic leukaemia (JMML) is a fatal, mixed myeloproliferative and myelodysplastic disorder of early childhood. A number of significant advances have been achieved in recent years. The present review will focus on the pathogenesis and management of JMML. Specific defects in the RAS signalling pathway, which make JMML cells hypersensitive to granulocyte-macrophage colony stimulating factor, are observed in at least two-thirds of patients with JMML: inactivation of NF1 or mutations in NRAS, KRAS2 or PTPN11. Allogeneic haematopoietic stem cell transplantation is currently the only treatment able to cure JMML, although the high postrelapse rate is of great concern. We also review molecularly targeted therapeutics that interfere with the disrupted activation of the RAS signal transduction pathway. PMID- 18422787 TI - Risk stratification and venous thromboprophylaxis in hospitalized medical and cancer patients. AB - Acute venous thromboembolism (VTE) is a serious and potentially fatal disorder, which often complicates the course of hospitalized medical patients. This is particularly true for carriers of malignant diseases. While the introduction of thromboprophylactic measures has probably affected the occurrence of postoperative VTE, there is an increasing awareness of the importance of medical conditions in determining thromboembolic events. Simple and clinically relevant risk assessment models are available to facilitate VTE risk assessment in hospitalized medical patients. Their validation in prospective studies is in progress. Randomized controlled clinical trials have consistently documented the efficacy of heparins and fondaparinux for prevention of VTE in hospitalized medical patients with a minimal haemorrhagic risk. Recognition of the incidence and clinical importance of thrombosis will probably encourage more widespread use of antithrombotic prophylaxis in medical patients and especially in some particular types of malignancies. PMID- 18422788 TI - Enzymes, detergents and skin: facts and fantasies. AB - In their raw state, enzymes of bacterial/fungal origin cause allergic reactions in the lung. Proteolytic enzymes also cause irritation to skin, eyes and the respiratory tract. For 40 years, encapsulated enzymes have been used worldwide in detergent products, especially laundry formulations, and have increasing importance due to biodegradability and functionality at low temperatures, offering environmental benefits. Uniquely to the U.K., for years it has been suggested that the inclusion of enzymes in such products leads to adverse skin reactions, including erythema, pruritus and exacerbation of eczema. In this review, we look at the facts, asking whether there is evidence that the hazards identified for enzymes translate into any risk for consumer health. By considering the actual exposures in consumer use and exaggerated product usage, it is concluded that the irritating and allergenic hazards of enzyme raw materials do not translate into a risk of skin reactions, either irritant or allergic. Investigations of numerous individuals with skin complaints attributed to laundry products demonstrate convincingly that enzymes were not responsible. Indeed, enzyme-containing laundry products have an extensive history of safe use. Thus, the supposed adverse effects of enzymes on skin seem to be a consequence of a mythology. The important practical lesson is that when primary or secondary care practitioners are presented with a skin complaint, it should not be dismissed as a result of using an enzyme-containing laundry product as the diagnosis will certainly lie elsewhere. Education for healthcare professionals could usefully be enhanced to take this on board. PMID- 18422789 TI - New insights into disease pathogenesis in crusted (Norwegian) scabies: the skin immune response in crusted scabies. AB - BACKGROUND: Crusted scabies is a rare and severely debilitating disease characterized by infestation of the skin with up to millions of Sarcoptes scabiei mites, high total IgG levels, extremely high total IgE levels, and the development of hyperkeratotic skin crusts that may be loose, scaly and flaky or thick and adherent. OBJECTIVES: To describe crusted scabies skin pathogenesis and identify markers associated with an inappropriate immune response leading to disease progression. PATIENTS/METHODS: Serial sections from skin biopsies obtained from two patients with severe crusted scabies were examined by immunohistochemistry for cell surface markers and inflammatory and regulatory cytokines. Concurrent levels of total B- and T-cell subsets and IgE, IgA, IgM, IgG and IgG subclasses were analysed in the blood. In addition antibody levels were recorded in a further 33 patients with crusted scabies and 14 patients with ordinary scabies. RESULTS: A predomination of infiltrating CD8+ T lymphocytes in the dermis was observed compared with minimal helper T lymphocytes (CD4+) and the absence of any B cells. The proportion of T and B lymphocytes and T-cell subsets in the blood of these patients were within normal ranges, indicating a selective movement of CD8+ T cells into the dermis. Furthermore, strong staining for the inflammatory cytokine interleukin-1 beta and anti-inflammatory cytokine transforming growth factor-beta1 was observed. Elevated levels of IgE, IgG, IgG1, IgG3 and IgG4 were recorded. CONCLUSIONS: Skin-homing cytotoxic T cells contribute to an imbalanced inflammatory response in the dermis of crusted scabies lesional skin. This, in combination with the lack of B cells, is contributing to the failure of the skin immune system to mount an effective response resulting in uncontrolled growth of the parasite. PMID- 18422790 TI - Regional spatial modeling of topsoil geochemistry. AB - Geographic information about the levels of toxics in environmental media is commonly used in regional environmental health studies when direct measurements of personal exposure is limited or unavailable. In this article, we propose a statistical framework for analyzing the spatial distribution of topsoil geochemical properties, including the concentrations of various toxicants. Due to the small-scale heterogeneity of most geochemical topsoil processes, direct measurements of the processes themselves only provide highly localized information; it is thus financially prohibitive to study the spatial patterns of these processes across a large region using traditional geostatistical analyses of point-referenced topsoil data. Instead, it is standard practice to assess geochemical patterns at a regional scale using point-referenced measurements collected in stream sediment because, unlike topsoil data, individual stream sediment geochemical measurements are representative of the surrounding area. We propose a novel multiscale soils (MSS) model that formally synthesizes data collected in topsoil and stream sediment and allows the richer stream sediment information to inform about the topsoil process, which in environmental health studies is typically more relevant. Our model accommodates the small-scale heterogeneity of topsoil geochemical processes by modeling spatial dependence at an aggregate resolution corresponding to hydrologically similar regions known as watersheds. We present an analysis of the levels of arsenic, a toxic heavy metal, in topsoil across the midwestern United States using the MSS model and show that this model has better predictive abilities than alternative approaches using more conventional statistical models for point-referenced spatial data. PMID- 18422791 TI - Estimation for the optimal combination of markers without modeling the censoring distribution. AB - In the time-dependent receiver operating characteristic curve analysis with several baseline markers, research interest focuses on seeking appropriate composite markers to enhance the accuracy in predicting the vital status of individuals over time. Based on censored survival data, we proposed a more flexible estimation procedure for the optimal combination of markers under the validity of a time-varying coefficient generalized linear model for the event time without restrictive assumptions on the censoring pattern. The consistency of the proposed estimators is also established in this article. In contrast, the inverse probability weighting (IPW) approach might introduce a bias when the selection probabilities are misspecified in the estimating equations. The performance of both estimation procedures are examined and compared through a class of simulations. It is found from the simulation study that the proposed estimators are far superior to the IPW ones. Applying these methods to an angiography cohort, our estimation procedure is shown to be useful in predicting the time to all-cause and coronary artery disease related death. PMID- 18422792 TI - Bayesian distributed lag models: estimating effects of particulate matter air pollution on daily mortality. AB - A distributed lag model (DLagM) is a regression model that includes lagged exposure variables as covariates; its corresponding distributed lag (DL) function describes the relationship between the lag and the coefficient of the lagged exposure variable. DLagMs have recently been used in environmental epidemiology for quantifying the cumulative effects of weather and air pollution on mortality and morbidity. Standard methods for formulating DLagMs include unconstrained, polynomial, and penalized spline DLagMs. These methods may fail to take full advantage of prior information about the shape of the DL function for environmental exposures, or for any other exposure with effects that are believed to smoothly approach zero as lag increases, and are therefore at risk of producing suboptimal estimates. In this article, we propose a Bayesian DLagM (BDLagM) that incorporates prior knowledge about the shape of the DL function and also allows the degree of smoothness of the DL function to be estimated from the data. We apply our BDLagM to its motivating data from the National Morbidity, Mortality, and Air Pollution Study to estimate the short-term health effects of particulate matter air pollution on mortality from 1987 to 2000 for Chicago, Illinois. In a simulation study, we compare our Bayesian approach with alternative methods that use unconstrained, polynomial, and penalized spline DLagMs. We also illustrate the connection between BDLagMs and penalized spline DLagMs. Software for fitting BDLagM models and the data used in this article are available online. PMID- 18422793 TI - Analysis of capture-recapture models with individual covariates using data augmentation. AB - I consider the analysis of capture-recapture models with individual covariates that influence detection probability. Bayesian analysis of the joint likelihood is carried out using a flexible data augmentation scheme that facilitates analysis by Markov chain Monte Carlo methods, and a simple and straightforward implementation in freely available software. This approach is applied to a study of meadow voles (Microtus pennsylvanicus) in which auxiliary data on a continuous covariate (body mass) are recorded, and it is thought that detection probability is related to body mass. In a second example, the model is applied to an aerial waterfowl survey in which a double-observer protocol is used. The fundamental unit of observation is the cluster of individual birds, and the size of the cluster (a discrete covariate) is used as a covariate on detection probability. PMID- 18422794 TI - A new method of providing pulsatile flow in a centrifugal pump: assessment of pulsatility using a mock circulatory system. AB - Previous studies have demonstrated the potential advantages of pulsatile flow as compared with continuous flow. However, to date, physiologic pumps have been technically complex and their application has therefore remained in the experimental field. We have developed a new type of centrifugal pump, which can provide pulsatile as well as continuous flow. The inner wall of a centrifugal pump is pulsed by means of a flexible membrane, which can be accurately controlled by means of either a hydraulic or pneumatic driver. The aim of this study was to assess the hydraulic behavior of the new pump in terms of surplus hemodynamic energy (SHE). We conducted experiments using a mock circulatory system including a membrane oxygenator. No differences were found in the pressure flow characteristics between the new pump and a conventional centrifugal pump, suggesting that the inclusion of the flexible membrane does not alter hydraulic performance. The value of SHE rose when systolic volume was increased. However, SHE dropped when the percentage of ejection time was reduced and also when the continuous flow (programmed by the centrifugal console) increased. Mean flow matched well with the continuous flow set by the centrifugal console, that is, the pulsatile component of the flow was exclusively controlled by the pulsatile console, and was therefore independent of the continuous flow programmed by the centrifugal console. The pulsatility of the new pump was approximately 25% of that created with a truly pulsatile pump. PMID- 18422795 TI - Antithrombogenic properties of a monopivot magnetic suspension centrifugal pump for circulatory assist. AB - The National Institute of Advanced Industrial Science and Technology (AIST) monopivot magnetic suspension centrifugal pump (MC105) was developed for open heart surgery and several weeks of circulatory assist. The monopivot centrifugal pump has a closed impeller of 50 mm in diameter, supported by a single pivot bearing, and is driven through a magnetic coupling to widen the fluid gap. Design parameters such as pivot length and tongue radius were determined through flow visualization experiments, and the effectiveness was verified in preliminary animal experiments. The maximum overall pump efficiency reached 18%, and the normalized index of hemolysis tested with bovine blood was as low as 0.0013 g/100 L. Animal experiments with MC105 were conducted in sheep for 3, 15, 29, and 35 days in a configuration of left ventricle bypass. No thrombus was formed around the pivot bearing except when the pump speed was reduced by 20% of normal operational speed, which reduced the pump flow by 40% to avoid inlet suction. Subsequently, the antithrombogenic design was verified in animal experiments for 5 weeks at a minimum rotational speed of greater than 1500 rpm and a minimum pump flow greater than 1.0 L/min; no thrombus formation was observed under these conditions. PMID- 18422796 TI - Mock circulatory system for the evaluation of left ventricular assist devices, endoluminal prostheses, and vascular diseases. AB - A new digital computer mock circulatory system has been developed in order to replicate the physiologic and pathophysiologic characteristics of the human cardiovascular system. The computer performs the acquisition of pressure, flow, and temperature in an open loop system. A computer program has been developed in Labview programming environment to evaluate all these physical parameters. The acquisition system was composed of pressure, flow, and temperature sensors and also signal conditioning modules. In this study, some results of flow, cardiac frequencies, pressures, and temperature were evaluated according to physiologic ventricular states. The results were compared with literature data. In further works, performance investigations will be conducted on a ventricular assist device and endoprosthesis. Also, this device should allow for evaluation of several kinds of vascular diseases. PMID- 18422797 TI - Uremic pruritus, cytokines, and polymethylmethacrylate artificial kidney. AB - Uremic pruritus is one of the common complications in long-term dialysis patients. Recently, researchers reported that immunohypothesis with high serum level of cytokines could be the cause of uremic pruritus. Polymethylmethacrylate (PMMA) artificial kidney (AK) has been reported to adsorb more serum cytokines than other high-flux AKs. In July 2006, 30 patients with severe uremic pruritus from 300 chronic hemodialysis (HD) patients in a single center entered this prospective study. Their dialyzers were changed to PMMA AK for 4 weeks. The severity of pruritus was evaluated every week using the results of a questionnaire (pruritus score). Laboratory assays including predialysis serum blood urea nitrogen (BUN), creatinine, beta2-microglobulin (beta2M), calcium, phosphate, intact parathyroid hormone (iPTH), total CO(2), ferritin, hematocrit, high-sensitivity C-reactive protein (hsCRP), IL-1beta, IL-2, IL-6, IL-18, tumor necrosis factor-alpha (TNF-alpha), Kt/V, and beta2M clearance were measured before and at the end of 4 weeks of PMMA AK use. PMMA AK was effective in reducing the pruritus score from 23.46 +/- 11.94 to 7.38 +/- 6.42 (P < 0.001). The effect of uremic pruritus relief appeared after 1 week of PMMA AK use. There were no significant differences in the laboratory assay results including predialysis serum BUN, Cr, beta2M, calcium, phosphate, calcium-phosphate product, iPTH, total CO(2), ferritin, hematocrit, hsCRP, IL-1beta, IL-2, IL-6, IL-18, TNF alpha, Kt/V, and beta2M clearance. The mechanism for the beneficial effect of PMMA AK on uremic pruritus remains to be determined. PMMA AK may be a useful adjuvant therapy in chronic HD patients with severe uremic pruritus. PMID- 18422798 TI - Development of watertight and bioabsorbable synthetic dural substitutes. AB - We have developed novel watertight and bioabsorbable synthetic dural substitutes. The substitutes were designed such that they had a three-layered structure, and each layer comprised a bioabsorbable copolymer prepared from L-lactide, glycolide, and epsilon-caprolactone. Various copolymers were synthesized, and appropriate compositions were selected for preparing the substitutes based on the results of leakage tests. Experimental substitutes that prevented the leakage of saline through the suture lines between the substitutes and dura mater were prepared. An in vitro study was performed in which the substitutes were sutured to porcine dura mater, and the amount of saline leakage was measured. It was demonstrated that leakage through the suture line could be completely suppressed by using the substitutes we developed along with favorable suturing techniques. PMID- 18422799 TI - Plasma exchange before surgery for left ventricular assist device implantation. AB - Left ventricular assist device (LVAD) implantation in end-stage heart failure patients is frequently associated with hemorrhagic complications requiring reoperation. The preoperative coagulopathic profile includes prolonged prothrombin time (PT), partial thromboplastin time (PTT), and bleeding time; platelet dysfunction; decreased coagulation factor activity; and increased inflammatory markers. We compare outcomes in LVAD patients treated with preoperative plasma exchange with concurrent, nonrandomized control patients. We reviewed data from 68 consecutive elective patients who received LVADs at our institution. Thirty-five received LVADs after preoperative plasma exchange (replacement of one plasma volume of fresh frozen plasma), and 33 received LVADs without plasma exchange. Groups were comparable in age, sex, body weight, New York Heart Association class, intra-aortic balloon pump insertion, cardiac index, pulmonary capillary wedge pressure, creatinine, total bilirubin, hemoglobin levels, PT, international normalized ratio, PTT, and platelet count. Early mortality was lower in the plasma exchange group (0% [0/35] vs. 18% [6/33], P = 0.026), and postoperative chest tube drainage decreased by 33% (P = not significant). Blood transfusion requirements were similar. Perioperative mortality decreased in patients treated with plasma exchange before LVAD implantation. PMID- 18422800 TI - Flow behavior within the 12-cc Penn State pulsatile pediatric ventricular assist device: an experimental study of the initial design. AB - Planar particle image velocimetry was used to explore the flow behavior of the newly designed 12-cc Penn State pneumatic pediatric assist pump. Wall shear maps complemented the velocity data. Bjork-Shiley Monostrut 17-mm mechanical heart valves were used in the inlet and outlet ports. In comparison with larger Penn State pumps, the 12-cc device is not only smaller but has reduced valve effective orifice areas and more highly angled valve ports. In contrast to results from the larger pumps, the flow field was highly three dimensional during early diastole with poorer penetration by the valve inlet jet. This led to a later start to a "wall washing" rotational pattern. A significant separation region, never before observed, was created upstream of the outlet valve leaflet during late diastole- effectively reducing the area and increasing the pressure drop through the valve. Wall shear maps suggest that regions of low shear might persist throughout the cycle at the bottom of the pump on the outlet side. An attempt to improve the flow field characteristics by exploring different valves, valve orientations and inlet valve angles, systolic/diastolic flow timing, and perhaps a larger outlet valve was planned. PMID- 18422801 TI - Characteristics of mechanical heart valve cavitation in a pneumatic ventricular assist device. AB - In previous studies, we investigated the mechanism of mechanical heart valve (MHV) cavitation and cavitation intensity with a nonsynchronized experiment system. Our group is currently developing a pneumatic ventricular assist device (PVAD), and in this study we investigated MHV cavitation intensity in the PVAD using a synchronized analysis of the cavitation images and the acoustic signal of cavitation bubbles. A 23-mm Medtronic Hall valve with an opening angle of 70 degrees was mounted in the mitral position of the PVAD after removing the sewing ring. A function generator provided a square signal, which used the trigger signal of the electrocardiogram R wave (ECG-R) mode of the control-drive console for circulatory support. This square signal was delayed by a delay circuit and was then used as the trigger signal for a pressure sensor and a high-speed video camera. The data were stored using a digital oscilloscope at a 1-MHz sampling rate, and then the pressure signal was band-pass filtered between 35 and 200 kHz using a digital filter. The band-pass filtered root mean squared (RMS) pressure and cavitation cycle duration were used as an index of cavitation intensity. The cavitation bubbles were concentrated at the valve stop, and the cavitation cycle duration and RMS pressure increased as the heart rate and driving pressure increased. At the low valve-closing velocity, bubble cavitation was observed near the valve stop. However, at the fast valve-closing velocity, cloud cavitation was observed. A high-frequency signal wave was generated when the bubbles collapsed. The cavitation cycle duration and RMS pressure increased as the valve-closing velocity increased linearly. PMID- 18422802 TI - Intra-aortic balloon pump-induced pulsatile flow reduces coagulative and fibrinolytic response to cardiopulmonary bypass. AB - The aim of this study is to evaluate if a simple intra-aortic balloon pump (IABP) induced pulsatile perfusion reduces activation of coagulative system during cardiopulmonary bypass (CPB). Ninety-six patients undergoing preoperative IABP were randomized to nonpulsatile CPB with IABP discontinued during cardioplegic arrest (Group A) or IABP-induced pulsatile CPB (Group B). White blood cells (WBC), hematocrit (Ht), platelets (PLTs), International Normalized Ratio (INR), fibrinogen, activated partial thromboplastin time (aPTT), antithrombin III (AT III) activity, and D-dimer were measured at the end of surgery (ES) and postoperatively. Chest drainage, need for reexploration, and transfusions were compared. Group B showed lower chest drainage (1st day P = 0.038; 2nd day P = 0.044), transfusions (P = 0.031), WBC (P < 0.05 at all time points), and INR (P < 0.05 at all time points), together with a higher Ht (P < 0.05 at ES, 12 h), platelets (P < 0.04 at all time points), fibrinogen (P < 0.05 at ES, 12 h, 24 h), and aPTT (P < 0.05 at all time points). AT-III activity lowered in Group A (P = 0.001 at ES, 12 h, 24 h), together with higher D-dimer levels (P < 0.05 at all time points). IABP-induced pulsatile perfusion ameliorates coagulative system activation following CPB. PMID- 18422803 TI - The value of clinical features in differentiating between viral, pneumococcal and atypical bacterial pneumonia in children. AB - OBJECTIVE: To evaluate the value of clinical features in differentiating between viral, pneumococcal and atypical bacterial pneumonia in children. DESIGN: A retrospective analysis of clinical signs and symptoms, supplemented with chest radiograph and serum procalcitonin data, in 101 children with community-acquired pneumonia. Viral and bacterial aetiology was studied prospectively by antibody assays, and pneumococcal infection was found in 18, atypical bacterial infection in 28 and viral infection alone in 22 cases. METHODS: Chest radiographs and serum procalcitonin were studied in all cases. Data on clinical signs and symptoms were retrospectively collected from the medical cards of the patients. RESULTS: Among symptoms, cough was present in 89% and fever (>37.5 degrees C) in 88% of the cases. Among physical signs, crackles were present in 49% and decreased breath sounds in 58%. No significant associations were found between any of the clinical signs or symptoms and the aetiology of pneumonia. In multivariate analyses, age over 5 years and serum procalcitonin over 1.0 ng/mL were the only independent predictors of bacterial aetiology, but no finding was able to screen between pneumococcal and atypical bacterial aetiology of infection. CONCLUSION: No clinical or radiological characteristic was helpful in the separation between viral, pneumococcal and atypical bacterial aetiology of community-acquired pneumonia (CAP) in children. PMID- 18422804 TI - A new technique for transumbilical insertion of central venous silicone catheters in newborn infants. AB - AIM: A new technique allowing placement of umbilical silicone venous catheters (USVC) is described and compared with percutaneous silicone venous catheters (PSVC). METHODS: Data were retrospectively recorded for 198 infants with USVC and 141 infants with PSVC. RESULTS: Overall rate of complications was low and comparable in both groups: thrombosis 1.2%, catheter-related sepsis 3.5% and mechanical obstruction 5%. CONCLUSION: A new device allows safe introduction of silicone catheters into the umbilical vein. PMID- 18422805 TI - Comparison of 2-h versus 3-h enteral feeding in extremely low birth weight infants, commencing after birth. AB - AIM: Immaturity is associated with problems in enteral nutrition of extremely low birth weight (ELBW) infants. Different time intervals between single feedings are used; however, no data are available to show a benefit of either regime. METHODS: In January 2001 enteral feeding regime was changed from 2-h to 3-h intervals. In a retrospective study charts were analysed for all ELBW infants during a period of 2 years prior (01/99-12/00) and after (08/01-07/03) changing the feeding regime. RESULTS: Forty-two in the 2-h group (gestational age 27 +/- 2.1, birth weight 797 +/-150) and 32 infants in 3-h (GA 26.9 +/- 1.8 weeks, BW 809 +/- 148 g) were included. Median (range) time until complete enteral feeding (26 (7 to 69) vs. 20 (12 to 58) days) was not statistical different. There were no differences with respect to enteral morbidity (NEC, abdominal surgery, feeding intolerance), length of stay (84 +/- 23 vs. 86 +/- 26 days), growth parameters or weight at discharge. Total duration of phototherapy and average length of continuous positive airway pressure (CPAP) support were significantly (p < 0.01) longer in the 3-h feeding group. CONCLUSION: Weight gain and time until complete enteral nutrition are similar in 2-h and 3-h feeding regimes. Data suggest an advantage of 2-h feedings concerning the length of CPAP and phototherapy. PMID- 18422806 TI - Exchange transfusion in infants with extreme hyperbilirubinemia: an experience from a developing country. AB - OBJECTIVE: To determine the clinical and epidemiological features of infants with extreme hyperbilirubinemia who require exchange transfusion (ET). STUDY DESIGN: Term and near-term infants admitted to a paediatric hospital over a 5-year period in La Paz, Bolivia, were included in the study if they met the following criteria: age less than 30 days; total serum bilirubin (TSB) levels > or =428 micromol/L (> or =25 mg/dL) on admission and if they had undergone an ET. RESULTS: Fifty-six infants were identified during the study period. Most of them were exclusively breastfed (98.2%). Median age at admission was 7 days. Weight loss of more than 12% since birth, as well as dehydration, was registered in more than one-third of cases. Extreme hyperbilirubinemia was considered as secondary to increased enterohepatic circulation (EHC) in most cases (73%). The mean TSB level on admission was 531.8 micromol/L (31.1 mg/dL). Fifteen infants (26.8%) showed signs of acute bilirubin encephalopathy (ABE). Early ABE was reversible after ET in most cases but all infants with advanced ABE developed severe kernicteric sequelae. Nine patients (16.1%) developed adverse events attributable to ET. CONCLUSIONS: Dehydration and weight loss in breastfed infants appeared to be an important factor associated with extreme hyperbilirubinemia and secondary brain damage during the first week of life. This may well be avoided if signs of ABE and its associated conditions are identified appropriately by follow-up programmes. PMID- 18422809 TI - Iron supplementation of iron-replete Indonesian infants is associated with reduced weight-for-age. AB - BACKGROUND: General iron supplementation to prevent iron deficiency in infants who are iron sufficient when starting supplementation may adversely affect their health. OBJECTIVE: A secondary analysis to explore the effect of iron supplementation on iron-replete (IR; Hb > or =113 g/L and S-ferritin > or =33 microg/L) or non-iron-replete 6-month-old Indonesian infants participating in a large, randomized trial on iron and zinc supplementation. RESULTS: Among the iron supplemented IR (Fe-IR, n = 80) infants S-ferritin was, compared to non-iron supplemented (NS) IR infants (NS-IR, n = 74), significantly higher (47.5 vs. 20.7 microg/L, p = 0.04), and S-zinc significantly lower (9.7 vs. 10.5 micromol/L, p = 0.04). Haemoglobin concentration (Hb) did not differ between the Fe-IR and NS-IR groups. Change in weight-for-age z-score (WAZ) from 6 to 12 months and mean WAZ at 12 months was lower in the Fe-IR group compared to the NS-IR group (-1.45 vs. 1.03, p < 0.001 and -1.97 vs. -1.60, p < 0.001, respectively). There was no difference in morbidity between groups. Iron supplementation of non-iron-replete infants increased Hb and S-ferritin, but did not affect S-zinc or anthropometrical indices. CONCLUSION: In our study, iron supplementation of IR infants affected WAZ adversely, whereas iron supplementation to non-iron-replete infants did not affect growth. These results support a cautious approach to iron supplementation of IR infants. PMID- 18422810 TI - Charcot-Marie-Tooth disease type 1B: marked phenotypic variation of the Ser78Leu mutation in five Italian families. AB - OBJECTIVES: To describe clinical, electrophysiological and genetic data of five unrelated Sicilian pedigrees harbouring a heterozygous Ser78Leu mutation in the myelin protein zero (MPZ) extracellular domain. MATERIALS AND METHODS: Clinical, electrophysiological and genetic findings of 16 patients were reported. Polymorphic markers flanking the coding sequence of MPZ gene were also analysed. RESULTS: A wide range of age at onset was observed in families 1 and 3, with a clinical heterogeneity, in terms of severity of the disease, within the same family (families 1 and 3), and among families. A markedly unsteady gait was a distinctive feature of many members of family 1. All patients in family 2 complained of severe cramps and painful paresthesia. Molecular genetic analysis showed that all affected subjects shared a common haplotype at three microsatellite loci D1S2858, D1S2624 and D1S484. CONCLUSIONS: Our study provides further evidence that phenotypic features of MPZ mutations can vary within and among different families. High frequency of Ser78Leu mutation in Sicily as well as the results of haplotype analyses suggest that the mutation may have been inherited from a common ancestor. PMID- 18422811 TI - Treatment with tumor necrosis factor inhibitors and intravenous immunoglobulin improves live birth rates in women with recurrent spontaneous abortion. AB - PROBLEM: The purpose of this study was to investigate whether treatment with tumor necrosis factor (TNF) inhibitors combined with intravenous immunoglobulin (IVIG) increases live birth rates among women with recurrent spontaneous abortion (RSA) concurrently treated with anticoagulants (AC). METHOD OF STUDY: Seventy five pregnancies in patients with a history of RSA were retrospectively evaluated. The population was divided into three groups: group I: 21 patients treated with AC (anticoagulants), group II: 37 patients treated with AC and IVIG, and group III: 17 patients treated with AC, IVIG and the TNF inhibitor Etanercept (Enbrel) or Adalimumab (Humira). In groups II and III, IVIG was administered at least once during the cycle of conception and/or at least once after a positive pregnancy test. In group III, either Adalimumab or Etanercept was administered by subcutaneous injection according to standard protocols. Statistical analysis of pregnancy outcome was performed using Fisher's exact test. RESULTS: Patient populations in the three treatment groups were similar in terms of age, past miscarriages, inherited thrombophilia and autoimmunity. The live birth rate was 19% (4/21) in group I, 54% (20/37) in group II, and 71% (12/17) in group III. There was significant improvement in pregnancy outcome in group II versus group I (P = 0.0127) and in group III versus group I (P = 0.0026). The live birth rate in group III compared to group II was not significantly different (P = 0.3723). Side effects of AC, IVIG and TNF inhibitor treatment were minimal in these patients, and no birth defects were identified in their offspring. CONCLUSION: In women with RSA, addition of either IVIG or a TNF inhibitor + IVIG to the AC regimen appears to improve live birth rates compared to the treatment with AC alone. The positive effect of IVIG and TNF inhibitor therapy on pregnancy outcome merits further study in prospective clinical trials. PMID- 18422812 TI - Alterations in RCAS1 serum concentration levels during the normal menstrual cycle and the lack of analogical changes in ovarian endometriosis. AB - PROBLEM: The selective suppression phenomenon of the cytotoxic immune response may be brought about by an increase in sRCAS1 blood serum concentration. Such an increase has been observed during cancer progression and decidualization. The aim of this study has been to evaluate alterations in RCAS1 blood serum concentration in women during normal menstrual cycle (NMC) phases and to compare the levels found with those in patients with ovarian endometriosis. METHOD OF STUDY: The sRCAS1 blood serum concentration level was determined for both 147 patients with ovarian endometriosis and a healthy control group. RESULTS: Fluctuations in sRCAS1 blood serum concentration levels correlating with the NMC phases were demonstrated. The highest level of sRCAS1 was found during menstruation and the lowest during the proliferative cycle phase. However, no such fluctuations correlating with menstrual cycle phases were observed in the patients with endometriosis. The level of sRCAS1 in patients with endometriosis remained almost constant. Moreover, the sRCAS1 blood serum concentration level has been shown to increase, as ovarian endometriosis progresses. CONCLUSION: RCAS1 blood serum concentration may be a useful indicator of the selective suppression of the immune cytotoxic cell phenomenon associated with the development of endometriosis. PMID- 18422813 TI - Placental fas/fas ligand expression in early pregnancy losses. AB - PROBLEM: The aim of this study was to compare the expression levels of Fas and Fas ligand (FasL) in first-trimester placentas obtained from spontaneous abortions in patients with antiphospholipid antibody syndrome (APS) or factor V (FV) Leiden mutation, compared with values in placentas from induced abortions in patients negative for these conditions. METHOD OF STUDY: We studied explants from 6- to 10-week-old placentas that had been prepared by collagenase digestion from 10 spontaneous abortions from APS-positive patients, nine spontaneous abortions in patients positive for FV Leiden mutation, and 10 induced abortions. All tissues were analyzed by flow cytometry for expression of Fas and FasL. RESULTS: Flow cytometric analysis showed that placental FasL expression was significantly lower in abnormal pregnancies than in normal ones. However, no such difference was observed for Fas expression. CONCLUSION: FasL on placental cells may be involved in the maintenance of immune privilege, thereby ensuring the safety and growth of placental tissues. Dysregulation of apoptotic mechanisms may play a critical role in spontaneous abortions. PMID- 18422814 TI - Irbesartan plus low-dose propranolol versus low-dose propranolol alone in cirrhosis: a placebo-controlled, double-blind study. AB - OBJECTIVES: Angiotensin II receptor antagonists have been shown to moderately lower portal pressure in some patients with cirrhosis but may have adverse effects on kidney function. This study aimed at comparing the effects of a combined treatment using irbesartan plus propranolol with propranolol monotherapy on portal pressure and kidney function in patients with cirrhosis. METHODS: Thirty-two patients were included (Child A/B/C: 13/18/1, etiology: 16 alcohol, 13 viral, 3 other; bilirubin 1.4 +/- 1.1 mg/dL, creatinine 0.86 +/- 0.20 mg/dL, baseline hepatic venous pressure gradient 18.7 +/- 5.3 mmHg). All patients received 20 mg propranolol b.i.d. Additionally, they randomly received either placebo (N = 15) or irbesartan (step-up dosage titration up to 300 mg/d, N = 17). Patients were followed at weekly intervals, re-evaluation of hepatic venous pressure gradient (HVPG) was performed after 8 wk. RESULTS: One patient in the propranolol/irbesartan group was excluded due to variceal bleeding. No other adverse events occurred. Portal pressure declined in both groups (propranolol/irbesartan group 19.6 +/- 1.5 mmHg to 16.6 +/- 1.2 mmHg, P= 0.037, propranolol/placebo group 17.8 +/- 1.1 mmHg to 15.1 +/- 1.2 mmHg, P= 0.019). Sodium excretion significantly increased in the propranolol/irbesartan group (from 122 +/- 20 mmol/d to 230 +/- 23 mmol/d, P= 0.045), but not in the propranolol/placebo group. CONCLUSIONS: Combination treatment of propranolol plus irbesartan is well tolerated in cirrhotic patients when titrating the angiotensin II antagonist in a step-up manner, and it increases sodium excretion in patients with compensated or moderately decompensated cirrhosis. Addition of irbesartan has no effect on portal pressure. PMID- 18422815 TI - Small intestinal bacterial overgrowth in patients suffering from scleroderma: clinical effectiveness of its eradication. AB - OBJECTIVES: After the skin, the gastrointestinal tract is the second most common target of systemic sclerosis (SSc). AIM: Our aims were to investigate orocecal transit time (OCTT) and the presence of small intestinal bacterial overgrowth (SIBO) in SSc as a cause of intestinal symptoms. METHODS: Fifty-five SSc patients and 60 healthy controls, sex and age matched, entered the study. Enrolled subjects completed a questionnaire for intestinal symptoms and a global symptomatic score (GSS) was calculated. OCTT and the presence of SIBO were assessed by a lactulose breath test (LBT). Patients with SIBO were treated with rifaximin 1,200 mg/day for 10 days. Finally, a second questionnaire and LBT were performed 1 month after the end of therapy. RESULTS: The prevalence of SIBO was higher in SSc patients compared with controls (30/54 vs 4/60, respectively, P < 0.001). OCTT was significantly slower in SSc patients compared with controls (150 min, 25-75th percentile 142.5-165 vs 105 min, 25-75th percentile 90-135, respectively, P < 0.001). In patients with SIBO, the median GSS score was 8 (25 75th percentile 3.25-10.75). Eradication of SIBO was achieved in 73.3% of patients, with a significant reduction of symptoms in 72.7% of them (GSS score 2, 25-75th percentile 1-3, P < 0.05). CONCLUSIONS: These data suggest that SIBO occurs more frequently in SSc patients than in controls. Intestinal symptoms in these patients may be related to this syndrome and its eradication seems useful to improve clinical features. OCTT is significantly delayed in SSc patients, suggesting an impairment of intestinal motility, a further risk factor for the development of SIBO. PMID- 18422816 TI - Relapses of inflammatory bowel disease during pregnancy: in-hospital management and birth outcomes. AB - BACKGROUND: There are few studies that describe the medical treatment and colitis response rates among patients with a severe relapse of inflammatory bowel disease (IBD) during pregnancy, and few studies of the effect of such a relapse on birth outcomes in these patients. OBJECTIVES: To describe the treatment and response rates of severe colitis in pregnancy, and to assess the effects of a severe relapse of colitis during pregnancy on birth outcomes. METHODS: We performed a case control study of pregnant patients with IBD hospitalized for a disease relapse at two large treatment centers between 1989 and 2001. Details of management of disease relapse and maternal and fetal outcomes were recorded. RESULTS: Eighteen patients (11 ulcerative colitis, 6 Crohn's disease, 1 indeterminate colitis), mean age 28.6 yr (range 19-38) formed the study group; 41 age-matched pregnant IBD patients without disease relapse formed the control group. Study patients were hospitalized at a mean of 15.9-wk gestation (range 8 35) for a mean of 10.4 days (range 3-31). All 18 patients received IV hydrocortisone (mean dose 199 mg/day) and 7 patients (39%) either continued taking or were commenced on immunomodulators: IV cyclosporine (5 patients) and azathioprine/6-MP (3 patients). Fifteen patients (83%) had a clinical response to these medical treatments, 3 patients required colectomy. There were significant differences between study and control groups in gestation period (35.0 wk vs 38.7 wk, respectively, P= 0.0001) and birth weight (2,001 g vs 3,018 g, respectively, P < 0.0001). CONCLUSIONS: Treatment with IV hydrocortisone and IV cyclosporine appears effective at inducing remission of colitis but their use must continue to be confined to severely ill patients being treated at specialized centers. Severe relapses of colitis during pregnancy increase the risk of preterm birth and low birth weight. PMID- 18422817 TI - Abdominal bloating in employed adults: prevalence, risk factors, and association with other bowel disorders. AB - BACKGROUND Bloating is common, but its significance as a marker of underlying disease has not been defined. AND AIMS: We report on risk factors for bloating, its relationship to physical activity and quality of life (QOL), and its predictive value for functional bowel disorders. METHODS: This is a cross sectional population-based study of 1,069 employees of the Veterans Affairs Black Hills Health Care System. The validated Bowel Disease Questionnaire was used to identify subjects with abdominal bloating and other bowel disorders. The association of bloating with QOL was assessed using the SF36 (Short-Form 36) questionnaire. Physical activity was assessed using the modified Baecke questionnaire. RESULTS: The response rate was 72% (723 of 1,069). Bloating was reported by 21% of all subjects (95% confidence interval [CI] 17.7-23.7), 64% with irritable bowel syndrome (IBS), 35% with non-IBS constipation, 23% with non IBS diarrhea, and 42% with dyspepsia. Functional bloating (i.e., bloating in the absence of other bowel disorders) was reported by 7% of subjects (95% CI 5.2 9.0). Of those with bloating, 28% had IBS, 25% non-IBS constipation, 8% non-IBS diarrhea, and 30% dyspepsia. The positive and negative predictive values of bloating in the diagnosis of functional bowel disorder were 66% and 87%, respectively. The only risk factors were smoking and high-dose aspirin. Bloating was not associated with physical activity. QOL on all subscales of SF36 was lower in subjects with bloating than those without bloating. CONCLUSIONS: Bloating is a common symptom in otherwise healthy adults, and is often associated with but not predictive of functional bowel disorders. Smoking and high-dose aspirin are associated with bloating while physical activity is not. PMID- 18422819 TI - Vitamin D status and bone density in recently diagnosed inflammatory bowel disease: the Manitoba IBD Cohort Study. AB - OBJECTIVES: Bone mineral density (BMD) is usually normal at the time of inflammatory bowel disease (IBD) diagnosis. The purpose of this study was to evaluate the role of vitamin D metabolism in recently diagnosed IBD. METHODS: Adult subjects with recently diagnosed IBD (median 4 yr) were recruited from the University of Manitoba IBD Research Registry into the Manitoba IBD Cohort Study. Baseline BMD and serum 25-hydroxy vitamin D (25OHD) were measured in a nested subgroup of 101 subjects of whom 94 had repeat BMD measurements 2.3 +/- 0.3 yr later. RESULTS: Only a minority (22 [21.8%]) of recently diagnosed IBD participants had optimal serum 25OHD levels (75 nmol/L or greater). Serum 25OHD was positively correlated with baseline BMD for the lumbar spine, total hip, and total body (all P < 0.05). MANOVA confirmed significant between-group differences in baseline T-scores when vitamin D status was categorized according to serum 25OHD quartile (P < 0.05). Gain in total body BMD between the baseline and follow up DXA scans was positively correlated with 25OHD (r = 0.20, P < 0.05). CONCLUSIONS: Poorer vitamin D status correlates with lower baseline BMD at all measurement sites and better vitamin D status is correlated with a gain in total body BMD. Early optimization of vitamin D may play an important role in preventing IBD-related bone disease. PMID- 18422820 TI - Association of TNFSF15 with Crohn's disease in Koreans. AB - OBJECTIVES: A recent genomewide association study from a Japanese population identified tumor necrosis factor superfamily member 15 (TNFSF15) as an inflammatory bowel disease gene. Previous studies have shown that expression of TNFSF15 was upregulated in macrophages and lymphocytes of the intestinal lamina propria of Crohn's disease (CD) patients. Here, we have tested four single nucleotide polymorphisms (SNPs) of TNFSF15 in Korean patients to determine whether the gene is associated with susceptibility to CD in a closely related population. METHODS: Four SNPs across TNFSF15 were genotyped in 380 patients with CD and 380 healthy controls. RESULTS: Carriers of three polymorphisms, including rs3810936, rs6478108, and rs7848647, showed statistically significant association with CD (adjusted OR [aOR] 2.81, 95% confidence interval [CI] 1.94-4.07, P= 4.4 x 10(-8); aOR 3.49, 95% CI 2.42-5.04, P= 2.7 x 10(-11); and aOR 3.49, 95% CI 2.42 5.03, P= 2.2 x 10(-11), respectively). Following haplotype analysis, homozygotes carrying two copies of the haplotype consisting of the risk alleles of those three SNPs showed statistically significant association with CD (aOR 5.39, 95% CI 3.19-9.10, P= 3.07 x 10(-10)). CONCLUSIONS: Our data support the hypothesis that the TNFSF15 genotypes play an important role in the pathogenesis of CD in Koreans. PMID- 18422821 TI - Prevalence of fibrosis and cirrhosis in chronic hepatitis B: implications for treatment and management. AB - OBJECTIVE: To document the prevalence and factors associated with severe fibrosis and cirrhosis in a large population of Asian chronic hepatitis B (CHB) patients. METHODS: Transient elastography was performed in unselected CHB patients. Liver stiffness score of <8.1 kPa was used as a cut-off for the presence of severe fibrosis or liver cirrhosis. RESULTS: 1315 patients were recruited, of which 951 (72%) were treatment-naive. Of these, 319 (34%) had severe fibrosis, with higher prevalence seen in males compared with females (39% vs 24% respectively, p < 0.01. Severe fibrosis was seen with increasing age from 20% in patients <25 years to 81% in those >65 years. Higher prevalence of severe fibrosis was seen in HBeAg(+) patients compared to HBeAg(-) patients age >45 years (58% vs 43% respectively, p = 0.03), in patients with HBV DNA levels >or=4 log compared with <4 log copies/ml (41% vs 27% respectively, p < 0.01), and in patients with stepwise increase of ALT levels (<0.5 x ULN vs 0.5-1 x ULN vs 1-2 x ULN; 11% vs 30% vs 48% respectively, p < 0.01). After multivariate analysis, gender, age and ALT levels were significant factors associated with severe fibrosis. Patients who received antiviral treatment had lower ALT, stiffness score and prevalence of cirrhosis compared to treatment-naive patients [25 vs 35 U/L (p < 0.01), 6.2 vs 6.7 kPa (p = 0.031) and 14% vs 22% (p = 0.008) respectively]. CONCLUSION: The overall prevalence of severe fibrosis in CHB patients was 34% with higher rates seen in older age groups, males, and in patients with higher ALT levels. PMID- 18422822 TI - Methamphetamine suppression and route of administration: precursor regulation impacts on snorting, smoking, swallowing and injecting. AB - AIMS: The route of drug administration affects risk for dependence and medical harm. This study examines whether routes used by methamphetamine treatment participants were impacted by a major drug suppression policy-federal regulation of the methamphetamine precursor chemicals ephedrine and pseudoephedrine. DESIGN: Autoregressive-integrated moving average (ARIMA) intervention time-series analysis. SETTING: California (1992-2004). INTERVENTIONS: Ephedrine single ingredient products regulation, implemented August 1995; ephedrine with other active medicinal ingredients regulation, implemented October 1996; pseudoephedrine products regulation, implemented October 1997. MEASUREMENTS: Monthly counts of non-coerced methamphetamine treatment admissions reporting snorting, smoking, swallowing or injecting. Findings After rising sharply, snorting, smoking, swallowing and injecting admissions dropped 50%, 43%, 26% and 26%, respectively, when the 1995 regulation was implemented. Snorting also dropped 38% at the time of the 1997 regulation. Snorting, swallowing and injecting remained at lower levels to the end of the study period. Smoking resurged (40%) at the time of the 1996 regulation and continued rising. CONCLUSIONS: Precursor regulation was associated with changes in the administration of methamphetamine. Injecting, the route with the greatest health risk, entered a long-term reduction. So, too, did snorting and swallowing, two routes with lower risk for dependence. In contrast, smoking, which has a relatively high risk for dependence, dropped, then rebounded and entered a long term rise. A possible explanation is that injecting, snorting and swallowing were largely linked with US domestic methamphetamine production, which has yet to recover from the regulations. While Mexican production, which was impacted only temporarily by the regulations and has supplanted domestic production, may have helped to diffuse smoking, a route with which it is historically correlated. PMID- 18422823 TI - The effect of proactively identifying smokers and offering smoking cessation support in primary care populations: a cluster-randomized trial. AB - AIMS: To establish whether proactively identifying all smokers in primary care populations and offering smoking cessation support is effective in increasing long-term abstinence from smoking. DESIGN: Cluster randomized controlled trial. SETTING: Twenty-four general practices in Nottinghamshire, randomized by practice to active or control intervention. PARTICIPANTS: All adult patients registered with the practices who returned a questionnaire confirming that they were current smokers (n = 6856). INTERVENTION: Participants were offered smoking cessation support by letter and those interested in receiving it were contacted and referred into National Health Service (NHS) stop smoking services if required. MEASUREMENTS: Validated abstinence from smoking, use of smoking cessation services and number of quit attempts in continuing smokers at 6 months. FINDINGS: Smokers in the intervention group were more likely than controls to report that they had used local cessation services during the study period [16.6% and 8.9%, respectively, adjusted odds ratio (OR) 2.09, 95% confidence interval (CI) 1.57 2.78], and continuing smokers (in the intervention group) were more likely to have made a quit attempt in the last 6 months (37.4% and 33.3%, respectively, adjusted OR 1.23, 95% CI 1.01-1.51). Validated point prevalence abstinence from smoking at 6 months was higher in the intervention than the control groups (3.5% and 2.5%, respectively) but the difference was not statistically significant (adjusted OR controlling for covariates: 1.64, 95% CI 0.92-2.89). CONCLUSIONS: Proactively identifying smokers who want to quit in primary care populations, and referring them to a cessation service, increased contacts with cessation services and the number of quit attempts. We were unable to detect a significant effect on long-term cessation rates, but the study was not powered to detect the kind of difference that might be expected. PMID- 18422824 TI - Genetic approaches to addiction: genes and alcohol. AB - AIMS: Alcoholism is a chronic relapsing disorder with an enormous societal impact. Understanding the genetic basis of alcoholism is crucial to characterize individuals' risk and to develop efficacious prevention and treatment strategies. METHODS: We examined the available scientific literature to provide an overview of different approaches that are being integrated increasingly to advance our knowledge of the genetic bases of alcoholism. Examples of genes that have been shown to influence vulnerability to alcoholism and related phenotypes are also discussed. RESULTS: Genetic factors account for more than 50% of the variance in alcoholism liability. Susceptibility loci for alcoholism include both alcohol specific genes acting either at the pharmacokinetic or pharmacodynamic levels, as well as loci moderating neuronal pathways such as reward, behavioral control and stress resiliency, that are involved in several psychiatric diseases. In recent years, major progress in gene identification has occurred using intermediate phenotypes such as task-related brain activation that confer the advantage of increased power and the opportunity of exploring the neuronal mechanisms through which genetic variation is translated into behavior. Fundamental to the detection of gene effects is also the understanding of the interplay between genes as well as genes/environment interactions. Whole Genome Association studies represent a unique opportunity to identify alcohol-related loci in hypothesis-free fashion. Finally, genome-wide analyses of transcripts and chromatin remodeling promise an increase in our understanding of the genome function and of the mechanisms through which gene and environment cause diseases. CONCLUSIONS: Although the genetic bases of alcoholism remain largely unknown, there are reasons to think that more genes will be discovered in the future. Multiple and complementary approaches will be required to piece together the mosaic of causation. PMID- 18422825 TI - Alcohol screening and brief counseling in a primary care hypertensive population: a quality improvement intervention. AB - AIMS: To determine the effect of an intervention to improve alcohol screening and brief counseling for hypertensive patients in primary care. DESIGN: Two-year randomized, controlled trial. SETTING/PARTICIPANTS: Twenty-one primary care practices across the United States with a common electronic medical record. INTERVENTION: To promote alcohol screening and brief counseling. Intervention practices received site visits from study personnel and were invited to annual network meetings to review the progress of the project and share improvement strategies. MEASUREMENTS: Main outcome measures included rates of documented alcohol screening in hypertensive patients and brief counseling administered in those diagnosed with high-risk drinking, alcohol abuse or alcohol dependence. Secondary outcomes included change in blood pressure among patients with these diagnoses. FINDINGS: Hypertensive patients in intervention practices were significantly more likely to have been screened after 2 years than hypertensive patients in control practices [64.5% versus 23.5%; adjusted odds ratio (OR) = 8.1; 95% confidence interval (CI) 1.7-38.2; P < 0.0087]. Patients in intervention practices diagnosed with high-risk drinking, alcohol abuse or alcohol dependence were more likely than those in control practices to have had alcohol counseling documented (50.5% versus 29.6%; adjusted OR = 5.5, 95% CI 1.3-23.3). Systolic (adjusted mean decline = 4.2 mmHg, P = 0.036) and diastolic (adjusted mean decline = 3.3 mmHg, P = 0.006) blood pressure decreased significantly among hypertensive patients receiving alcohol counseling. CONCLUSIONS: Primary care practices receiving an alcohol-focused intervention over 2 years improved rates of alcohol screening for their hypertensive population. Implementation of alcohol counseling for high-risk drinking, alcohol abuse or alcohol dependence also improved and led to changes in patient blood pressures. PMID- 18422828 TI - Women's gender role orientation predicts their drinking patterns: a follow-up study of Czech women. AB - AIMS: Evaluation of the hypothesis that women's non-traditional gender role orientation contributes to drinking patterns typical for men. DESIGN: A two-wave prospective study with data collected in 1992 and 1997. SETTING: The data reflect Czech women's changing gender role orientation and their drinking patterns during a historical period of post-totalitarian societal transformation. PARTICIPANTS: A representative cohort of 497 Prague women aged 30-59 years in 1997. MEASUREMENTS: Face-to-face interview data on drinking patterns and individually collected original questionnaire on gender role orientation. FINDINGS: An analysis of the principal components of the gender role orientation questionnaire has led to four components, designated as egalitarianism, liberalism, feminism and hedonism. Constructed role orientation scales had Cronbachs's alpha reliabilities ranging from 0.57 to 0.74. With possible confounders controlled (thanks mainly to the prospective design), non-traditional gender role orientation components assessed in 1992 predicted the usual quantities of alcohol women have consumed per occasion in 1997, as well as three hazardous drinking patterns (occasional use of > or = 96 g alcohol, usual use of > or = 48 g and daily intake of > or = 40 g). Specifically, women's usual quantity per occasion and occasional use of > or = 96 g were predicted by egalitarianism and hedonism, and hedonism predicted usual use of > or = 48 g as well as average daily intake of > or = 40 g ethanol. CONCLUSIONS: Women's gender role orientation can be associated with their drinking patterns with non-traditional gender role identification being associated with greater likelihood of hazardous drinking. PMID- 18422826 TI - Alcohol measurement methodology in epidemiology: recent advances and opportunities. AB - AIM: To review and discuss measurement issues in survey assessment of alcohol consumption for epidemiological studies. METHODS: The following areas are considered: implications of cognitive studies of question answering such as self referenced schemata of drinking, reference period and retrospective recall, as well as the assets and liabilities of types of current (e.g. food frequency, quantity-frequency, graduated frequencies and heavy drinking indicators) and life time drinking measures. Finally we consider units of measurement and improving measurement by detailing the ethanol content of drinks in natural settings. RESULTS AND CONCLUSIONS: Cognitive studies suggest inherent limitations in the measurement enterprise, yet diary studies show promise of broadly validating methods that assess a range of drinking amounts per occasion; improvements in survey measures of drinking in the life course are indicated; attending in detail to on- and off-premise drink pour sizes and ethanol concentrations of various beverages shows promise of narrowing the coverage gap plaguing survey alcohol measurement. PMID- 18422829 TI - Long-term effects of heroin-assisted treatment in Germany. AB - AIMS: Trials in Switzerland, the Netherlands and Spain have found that heroin assisted treatment (HAT) as maintenance treatment for opioid-dependent patients reduces illicit drug use. A German trial also found diamorphine treatment to be superior to methadone treatment. The present study describes the association between 2 years of heroin treatment and improvements in health and social stabilization, as well as illicit drug use. DESIGN: A prospective cohort study design. PARTICIPANTS: A total of 515 patients were assigned to diamorphine treatment; 278 patients remained in the study treatment for the entire period of 24 months (54.8%). MEASUREMENTS: The results on physical (Opiate Treatment Index Health Symptoms Scale) and mental (Symptom Checklist 90-Revised Global Severity Index) health and illicit drug use (number of days with drug use within the last month-European Addiction Severity Index) were examined by repeated-measures analyses. FINDINGS: Symptoms of physical (Pillai's trace = 0.837, df = 4, P < 0.001) and mental health (Pillai's trace = 0.450, df = 4, P < 0.001) improved during treatment. Street heroin use declined rapidly (Pillai's trace = 0.836, df = 4, P < 0.001), as did cocaine use (Pillai's trace = 0.280, df = 4, P < 0.001). CONCLUSIONS: HAT is associated with improvements in mental and physical health in the long term. PMID- 18422827 TI - Critical issues in the treatment of hepatitis C virus infection in methadone maintenance patients. AB - AIMS: Hepatitis C virus (HCV) infection is a common chronic complication of injection drug use. Methadone maintenance programs contain large numbers of patients infected with HCV. This paper reviews HCV infection with emphasis on the medical care of HCV-infected, or HCV and human immunodeficiency virus co infected, patients on methadone or buprenorphine maintenance. METHODS: Literature searches using PubMed, PsycINFO and SocINDEX were used to identify papers from 1990-present on antiviral therapy for HCV in methadone maintenance patients and on liver transplantation in methadone maintenance patients. RESULTS: Injection drug use is the most significant risk factor for HCV infection in most western countries. The prevalence of HCV antibody is high in injection drug users (53 96%) and in patients enrolled in methadone maintenance programs (67-96%). Studies of antiviral therapy for HCV in methadone maintenance patients show rates of sustained virological response (SVR), defined as negative HCV-RNA 24 weeks after the end of treatment, of 28-94%. In studies with contrast groups, no significant differences in SVR between methadone and contrast groups were found. Excellent completion rates of antiviral therapy (72-100%) were found in five of six studies. There are many barriers to methadone maintenance patients' receiving antiviral therapy, and research on overcoming barriers is discussed. Liver transplantation has been successful in methadone maintenance patients but has not been utilized widely. CONCLUSION: High quality medical care for all aspects of HCV infection can be provided to methadone maintenance patients. The literature supports the effectiveness of such services, but the reality is that most patients do not receive them. PMID- 18422830 TI - Distinguishing signs of opioid overdose and indication for naloxone: an evaluation of six overdose training and naloxone distribution programs in the United States. AB - AIMS: This study assessed overdose and naloxone administration knowledge among current or former opioid abusers trained and untrained in overdose-response in the United States. DESIGN AND PARTICIPANTS: Ten individuals, divided equally between those trained or not trained in overdose recognition and response, were recruited from each of six sites (n = 62). SETTING: US-based overdose training and naloxone distribution programs in Baltimore, San Francisco, Chicago, New York and New Mexico. MEASUREMENTS: Participants completed a brief questionnaire on overdose knowledge that included the task of rating 16 putative overdose scenarios for: (i) whether an overdose was occurring and (ii) if naloxone was indicated. Bivariate and multivariable analyses compared results for those trained to untrained. Responses were also compared to those of 11 medical experts using weighted and unweighted kappa statistics. FINDINGS: Respondents were primarily male (72.6%); 45.8% had experienced an overdose and 72% had ever witnessed an overdose. Trained participants recognized more opioid overdose scenarios accurately (t(60) = 3.76, P < 0.001) and instances where naloxone was indicated (t(59) = 2.2, P < 0.05) than did untrained participants. Receipt of training and higher perceived competency in recognizing signs of an opioid overdose were associated independently with higher overdose recognition scores. Trained respondents were as skilled as medical experts in recognizing opioid overdose situations (weighted kappa = 0.85) and when naloxone was indicated (kappa = 1.0). CONCLUSIONS: Results suggest that naloxone training programs in the United States improve participants' ability to recognize and respond to opioid overdoses in the community. Drug users with overdose training and confidence in their abilities to respond may effectively prevent overdose mortality. PMID- 18422831 TI - The pharmacology of the endocannabinoid system: functional and structural interactions with other neurotransmitter systems and their repercussions in behavioral addiction. AB - Addiction is a chronic, recurring and complex disorder. It is characterized by anomalous behaviors that are linked to permanent or long-lasting neurobiological alterations. Furthermore, the endocannabinoid system has a crucial role in mediating neurotransmitter release as one of the main neuromodulators of the mammalian central nervous system. The purpose of the present review is to instruct readers about the functional and structural interactions between the endocannabinoid system and the main neurotransmitter systems of the central nervous system in the context of drug addiction. With this aim, we have systematically reviewed the main findings of most of the existing literature that explores cross-talk in the five brain areas that are most traditionally implicated in addiction: amygdala, prefrontal cortex, nucleus accumbens, hippocampus and ventral tegmental area (VTA). The neurotransmission systems influenced by the pharmacology of the endocannabinoid system in these brain areas, which are reviewed here, are gamma-aminobutyric acid (GABA), glutamate, the main biogenic amines (dopamine, noradrenaline and serotonin), acetylcholine and opioids. We show that all of these neurotransmitter systems can be modulated differentially in each brain area by the activation or deactivation of cannabinoid CB1 brain receptors. Specifically, most of the studies relate to the hippocampus and nucleus accumbens. Moreover, the neurotransmitter with the fewest number of related studies is acetylcholine (excepting in the hippocampus), whereas there is a large number that evaluates GABA, glutamate and dopamine. Finally, we propose a possible interpretation of the role of the endocannabinoid system in the phenomenon of addiction. PMID- 18422832 TI - The endocannabinoid system: emotion, learning and addiction. AB - The identification of the cannabinoid receptor type 1 (CB1 receptor) was the milestone discovery in the elucidation of the behavioural and emotional responses induced by the Cannabis sativa constituent Delta(9)-tetrahydrocannabinol. The subsequent years have established the existence of the endocannabinoid system. The early view relating this system to emotional responses is reflected by the fact that N-arachidonoyl ethanolamine, the pioneer endocannabinoid, was named anandamide after the Sanskrit word 'ananda', meaning 'bliss'. However, the emotional responses to cannabinoids are not always pleasant and delightful. Rather, anxiety and panic may also occur after activation of CB1 receptors. The present review discusses three properties of the endocannabinoid system as an attempt to understand these diverse effects. First, this system typically functions 'on-demand', depending on environmental stimuli and on the emotional state of the organism. Second, it has a wide neuro-anatomical distribution, modulating brain regions with different functions in responses to aversive stimuli. Third, endocannabinoids regulate the release of other neurotransmitters that may have even opposing functions, such as GABA and glutamate. Further understanding of the temporal, spatial and functional characteristics of this system is necessary to clarify its role in emotional responses and will promote advances in its therapeutic exploitation. PMID- 18422833 TI - Modulation of rat cerebellum oxidative status by prolonged red wine consumption. AB - A number of studies support the view that wine polyphenols can reinforce the endogenous antioxidant system by reducing ethanol (EtOH)-induced neuronal oxidative damage. Herein, we have investigated the effects of prolonged red wine (RW) consumption on several biomarkers of redox status in the cerebellum, a brain region highly vulnerable to the noxious effects of EtOH. Adult male Wistar rats were given RW with an EtOH concentration adjusted to 20% for 6 months, and the results were compared with those obtained in EtOH-treated (20%) and pair-fed control (PFC) animals. Malondialdehyde (MDA) and glutathione levels, and the activities of antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR), glutathione-S-transferase (GST) and selenium dependent glutathione peroxidase (Se-GPX) were estimated in cerebellum homogenates. Chronic RW ingestion resulted in diminished MDA and reduced glutathione levels in cerebellar tissue. Moreover, RW-treated rats had a significant decrease in SOD, GR and GST activities but presented an increase in the activity of Se-GPX compared with animals from EtOH and PFC groups. In contrast, CAT activity was not altered by RW and EtOH intakes. Taken together, these findings show that prolonged consumption of RW markedly modifies cerebellum redox status probably due to its high content of polyphenols. PMID- 18422834 TI - Baclofen reduces ethanol intake in high-alcohol-drinking University of Chile bibulous rats. AB - ABSTRACT Treatment with gamma-aminobutiric acid (GABA(B)) receptor agonist, +/ baclofen, has been shown to reduce ethanol intake in selectively bred Sardinian alcohol-preferring rats. The general goal of the present study was to characterize the high ethanol consumption high-alcohol-drinking University of Chile bibulous (UChB) rats with regard to the anti-alcohol effect of GABA(B) receptor stimulation. UChB rats were treated with the more active enantiomer of baclofen [R(+)-baclofen; at a dose of 1.0, 2.0 or 3.0 mg/kg] administered intraperitoneally once daily for four consecutive days or a single dose. When comparing ethanol and saccharin consumption in a free-choice regimen with unlimited access 24 hours/day, the dose of baclofen required to attenuate ethanol consumption significantly was 1.0 mg/kg administered once a day for three consecutive days while the dose that was sufficient to affect saccharin consumption significantly was 2.0 mg/kg, indicating that baclofen was more potent in reducing ethanol intake by UChB rats than reducing saccharin consumption. The reduction of ethanol or saccharin intake can not be attributed to baclofen induced motor impairment, since baclofen (1.0, 2.0 or 3.0 mg/kg) did not alter spontaneous locomotor activity in UChB rats. Baclofen dose-dependently suppressed the motor activity stimulated by ethanol administration, a phenomenon mediated by activation of the mesolimbic dopamine system. In conclusion, these results showed that the activation of GABA(B) receptor by R(+)-baclofen reduced ethanol and saccharin consumption, as well as ethanol-induced motor stimulation, implicating the GABA(B) receptor in the neural substrates mediating effects that sustain voluntary ethanol in take in UChB rats. PMID- 18422835 TI - Abnormalities in cortical and transcallosal inhibitory mechanisms in subjects at high risk for alcohol dependence: a TMS study. AB - Central nervous system (CNS) hyperexcitability and a resulting state of behavioral undercontrol are thought to underlie the vulnerability to early-onset alcohol dependence (AD). The aim of this study was to explore the differences in the functioning of cortical inhibitory systems, utilizing transcranial magnetic stimulation (TMS), in subjects at high risk (HR) and low risk (LR) for AD and to examine the relationship between CNS inhibition and behavioral undercontrol. Right-handed HR (n = 15) and LR (n = 15) subjects, matched for age, gender, height, weight and education, were assessed for psychopathology and family history of alcoholism using the Semi-Structured Assessment for the Genetics of Alcoholism and the Family Interview for Genetic Studies. Following single-pulse TMS, an electromyogram recorded from the right opponens pollicis muscle was used to measure the silent periods at different stimulus intensities. HR subjects had significantly shorter contralateral and ipsilateral (iSP) silent periods and a relatively higher prevalence of 'absent' iSP. They had significantly higher mean externalizing symptoms scores (ESS) than LR subjects, and there was a significant negative correlation between iSP duration and ESS. These preliminary findings suggest that HR subjects have relative impairments in corticocortical and transcallosal inhibitory mechanisms. The consequent state of CNS hyperexcitability may be etiologically linked to the excess of externalizing behaviors observed in this population, which is thought to be a predisposition to a higher risk of developing early-onset alcoholism. PMID- 18422836 TI - Ethanol enhances GABAergic transmission onto dopamine neurons in the ventral tegmental area of the rat. AB - BACKGROUND: Activation of the dopaminergic (DA) neurons of the ventral tegmental area (VTA) by ethanol has been implicated in its rewarding and reinforcing effects. At most central synapses, ethanol generally increases inhibitory synaptic transmission; however, no studies have explored the effect of acute ethanol on GABAergic transmission in the VTA. METHODS: Whole-cell patch clamp recordings of inhibitory postsynaptic currents (IPSCs) from VTA-DA neurons in midbrain slices from young rats. RESULTS: Acute exposure of VTA-DA neurons to ethanol (25 to 50 mM) robustly enhanced GABAergic spontaneous and miniature IPSC frequency while inducing a slight enhancement of spontaneous IPSC (sIPSC) amplitude. Ethanol (50 mM) enhanced paired-pulse depression of evoked IPSCs, further suggesting enhanced GABA release onto VTA-DA neurons. The frequency of sIPSCs was suppressed by the GABA(B) agonist, baclofen (1.25 microM) and enhanced by the antagonist, SCH50911 (20 microM); however, neither appeared to modulate or occlude the effects of ethanol on sIPSC frequency. CONCLUSIONS: The present results indicate that ethanol increases postsynaptic GABA(A) receptor sensitivity, enhances action potential-independent GABA release onto VTA-DA neurons, and that this latter effect is independent of GABA(B) auto-receptor inhibition of GABA release. PMID- 18422838 TI - The tachykinin receptor 3 is associated with alcohol and cocaine dependence. AB - BACKGROUND: A broad region on chromosome 4q was previously linked to the phenotype of alcohol dependence in the Collaborative Study on the Genetics of Alcoholism sample. A strong positional candidate gene was identified within this region: tachykinin receptor 3 gene (TACR3), which encodes tachykinin receptor 3 (NK3R), the receptor for the tachykinin 3 (neurokinin B) peptide. Pharmacological studies have provided evidence that the administration of NK3R agonists attenuates the intake of alcohol and NK3R can also mediate the acute and chronic behavioral effects of cocaine. METHODS: Thirty SNPs were genotyped throughout TACR3. Family based association analysis was performed in 219 European American families to detect an association with alcohol dependence. Subsequent analyses were performed to evaluate the evidence of association with other definitions of alcohol dependence as well as cocaine dependence. RESULTS: Seven of the 9 SNPs in the 3' region of TACR3 provided significant evidence of association with alcohol dependence (p A, 244G > A, 442C > A, 489G > A, 569G > A, 665G > A, and 950C > T). A transient expression study suggested that the 244G > A, 569G > A, and 950C > T SNPs are enzyme-inactivating mutations. CONCLUSION: This study identified 24 SNPs in the FUT2 gene, of which 7 were novel. The frequencies of alleles and genotypes were determined in Ovambos, Turks, and Mongolians. The allelic composition of each population was similar to those of geographically closer populations. PMID- 18422844 TI - Asymptomatic infection by Leishmania infantum in blood donors from the Balearic Islands (Spain). AB - BACKGROUND: Visceral leishmaniasis (VL) caused by Leishmania infantum is a zoonotic disease endemic throughout the Mediterranean basin. The existence of asymptomatic human infection entails the risk of transmission by blood transfusion. STUDY DESIGN AND METHODS: The prevalence of Leishmania infection was studied in 1437 blood donors from the Balearic Islands (Majorca, Formentera, and Minorca) using immunologic (Western blot [WB] and delayed-type hypersensitivity [DTH]), parasitologic (culture), and molecular (nested polymerase chain reaction [PCR]) methods. In addition, the efficiency of leukoreduction by filtration to remove the parasite was tested by nested PCR in the red blood cell (RBC) units. RESULTS: Leishmania antibodies were detected in 44 of the 1437 blood donors tested (3.1%). A sample of 304 donors from Majorca was selected at random. L. infantum DNA was amplified in peripheral blood mononuclear cells (PBMNCs) in 18 of the 304 (5.9%), and cultures were positive in 2 of the 304 (0.6%). DTH was performed on 73 of the 304 donors and was positive for 8 of them (11%). Of the 18 donors with positive L. infantum nested PCR, only 2 were seropositive. All the RBC samples tested (13 of 18) from donors with a positive PBMNC nested PCR yielded negative nested PCR results after leukodepletion. CONCLUSIONS: Cryptic Leishmania infection is highly prevalent in blood donors from the Balearic Islands. DTH and L. infantum nested PCR appear to be more sensitive to detect asymptomatic infection than the serology. The use of leukodepletion filters appears to remove parasites from RBC units efficiently. PMID- 18422845 TI - New mutations on platelet GPIIb in Sub-Saharan African populations revealed by genotyping discrepancies. AB - BACKGROUND: Previous studies of platelet allele frequencies in Sub-Saharan African populations enabled us to identify discrepancies in HPA-3 typing, suggesting the presence of new mutations and of a greater polymorphism than so far described in other populations. OBJECTIVES: To analyze these discrepancies and to assess the factors leading to potential alloimmunization in these populations. MATERIALS AND METHODS: SAMPLES: Maternal samples from a Beninese woman following in utero death and panels of blood donors from Benin, Cameroon, Congo, and Pygmies from Central Africa. TECHNIQUES: Genotyping was performed using polymerase chain reaction-restriction fragment length polymorphism (PCR RFLP), PCR-sequence specific primers (PCR-SSP) and sequencing techniques. RESULTS: Three new mutations were found on GPIIb gene: exon 26 a) 2614C>A situated between HPA-3 and HPA-9w, b) 2645C>T downstream of HPA-3, c) intron 26 IVS26+89G>A. These mutations may lead to discrepant DNA typing results, due either to a localization in the complementary sequence recognized by the primer or to the appearance of a new enzyme restriction site. Furthermore, a bilateral linkage << deletion (Delta9 bp) intron 21 and the HPA-3b allele (exon 26) >> found in Caucasian, Asian, and Oceanian populations is not found in African populations, suggesting that its appearance was prior to HPA-3. CONCLUSION: Three new mutations have been identified, two of them potentially immunogenic through their position. Furthermore, the polymorphism found on intron 26, localized in the complementary sequence of the PCR primer, may lead to a false typing assignation. It is therefore important to diversify techniques, both genomic (PCR RFLP and PCR-SSP), and proteomic monoclonal antibody-specific immobilization of platelets antigen (MAIPA) to ensure accurate HPA antigenic system typing. PMID- 18422846 TI - Prospective evaluation of a transfusion policy of D+ red blood cells into D- patients. AB - BACKGROUND: Although D- patients should receive red blood cells (RBCs) from D- donors, the scarcity of D- blood components in certain situations makes the transfusion of D+ RBCs unavoidable. Therefore it is recommended that guidelines be developed in order to standardize transfusion policy in these scenarios. STUDY DESIGN AND METHODS: We have prospectively evaluated a policy for the use of D+ RBCs in 905 D- patients. The amount of D- RBCs saved as well as the incidence of hemolytic reactions and anti-D alloimmunization were assessed. RESULTS: 554 patients received D- RBCs while 351 received a total of 1032 D+ RBCs, all of them within our criteria for the acceptable use of D+ RBCs. This strategy allowed us to save 25.6 percent of D- RBCs (1032 out of 4024 RBCs requested). No hemolytic reactions were reported. The incidence of alloimmunization was 21.4 percent. Most patients who developed anti-D did so within the first 2 or 4 RBCs transfused (64% after the first 2 RBCs transfused and 88% after the first 4). In multivariate analysis the age of less than 77 years was the only predictor for alloimmuization (HR = 2.48 [95% CI = 1.21-3.81]; p = 0.014). CONCLUSION: The use of D+ RBCs in selected D- patients does not induce adverse reactions and allows the saving of a significant number of D- RBCs. PMID- 18422847 TI - PRISM hepatitis B surface antigen detection of hepatits B virus minipool nucleic acid testing yield samples. AB - BACKGROUND: Hepatitis B virus (HBV) residual risk has been estimated at 1:63,000 1:205,000 and introduction of more sensitive serological tests and nucleic acid testing (NAT) would reduce that risk. Sensitivity of the recently licensed Abbott PRISM hepatitis B surface antigen (HBsAg) CLIA and minipool (MP) HBV NAT has been described as comparable and thus the need for HBV NAT has not been compelling. In this study, eight samples identified as yield samples with MP HBV NAT were tested using the PRISM test. STUDY DESIGN AND METHODS: Seven samples were identified using the Roche COBAS AmpliScreen HBV test and one additional sample was obtained from the clinical trial for the Roche cobas TaqScreen MPX test. Each of these samples was reactive by MP HBV NAT and nonreactive for HBsAg using one of three licensed enzyme immunoassay (EIA) tests. After licensure of the PRISM HBsAg, aliquots were tested with this assay, and DNA quantitation and genotyping were repeated where sample volume permitted. RESULTS: Three samples (2000, 2300, and 61,000 copies/mL) produced reactive results with PRISM. Four samples with viral loads less than 300 copies per mL produced nonreactive results. One sample, originally quantitated at 37,000 copies per mL (but 3850 copies/mL in repeat testing) was also nonreactive by PRISM. Genotyping of this sample indicated a type C genotype with no mutations. CONCLUSION: Adding serological sensitivity of PRISM CLIA reduced the NAT yield from the original 1: 385,555 to 1:610,488. However, MP HBV NAT still provides additional sensitivity over CLIA, even for a donation with a viral load of almost 4000 copies per mL. PMID- 18422848 TI - Loss of integrity of umbilical cord blood unit freezing bags: description and consequences. AB - BACKGROUND: Umbilical cord blood (UCB) is now a commonly used resource for hematopoietic stem cell (HSC) transplantation; great effort has been put forth in standardizing protocols for processing, storage, and testing of UCB units. Because UCB units are selected on an individual basis to maximize the chance of engraftment, loss of container integrity may have adverse effects on patient outcome. STUDY DESIGN AND METHODS: All bag breaks involving UCB units thawed for transplantation at our institution between January 1, 2000, and May 31, 2006, were identified. Information on various laboratory variables and the clinical consequences of UCB bag breaks was obtained from the deviation database of the Clinical Cell Therapy Laboratory (CCTL). Patient medical charts were reviewed for infusion-related data. RESULTS: The incidence of bag breaks over a 6 1/2-year period was 3.5 percent. A majority of cases of loss of container integrity occurred in units that had been cryopreserved for more than 2 years (75%) and resulted in minimal loss of product. There were no significant decreases in quantity or quality of UCB, as determined by various quality control tests; no adverse clinical outcomes related to receiving a broken UCB unit were noted except increased antibiotic usage. CONCLUSION: There was a relatively low incidence of UCB bag breaks in this study that did not result in significant loss of UCB or adverse clinical outcomes. With the FDA considering licensure of UCB for hematopoietic reconstitution, improvement in container design and possibly guidelines limiting length of storage will likely be addressed in detail. PMID- 18422849 TI - Detailed immunophenotypic characterization of different major and minor subsets of peripheral blood cells in patients with paroxysmal nocturnal hemoglobinuria. AB - BACKGROUND: Paroxysmal nocturnal hemoglobinuria (PNH) is characterized by a deficient expression of glycosylphosphatidylinositol-anchored proteins (GPI-APs), due to somatic mutations of the phosphatidylinositolglycan complementation Class A (PIG-A) gene. STUDY DESIGN AND METHODS: In this study, the expression of a high number of GPI-APs on different subsets of peripheral blood (PB) cells from 14 PNH patients and their potential association with underlying genetic abnormalities has been analyzed. RESULTS: This study confirms the existence of variable patterns of expression of different GPI-APs on both major and minor PB-cell subsets from PNH patients. The size of the PNH clone within PB neutrophils and monocytes was systematically higher than that of other cell populations. Genetic changes were detected in the PIG-A gene in 5 of 13 cases analyzed. Interestingly, the reactivity for many GPI-APs was significantly higher on different subsets of normal PB cells from PNH patients than those observed on healthy volunteers. CONCLUSION: The best combination of markers for the diagnostic screening of PNH would include evaluation of CD14 on monocytes and of CD16 on neutrophils, although further analysis of CD55 and CD59 expression may contain additional clinically useful information. Clear association between the genetic changes detected in the PIG-A gene in 5 of 13 cases analyzed, and the phenotypic profile of PNH cells has not been found. Additionally, an abnormally higher expression of several GPI-APs among normal residual cells from PNH patients in comparison to healthy donors was observed, suggesting that factors other than the PIG-A mutation could determine the phenotypic profile of PB cells in PNH. PMID- 18422850 TI - Noninvasive fetal RHD genotyping by maternal plasma with capillary electrophoresis. AB - BACKGROUND: Recently, a more accurate and reliable screening test has been investigated for noninvasive prenatal fetal RHD genotyping from D- women. The objective of this study was to perform the new method of noninvasive fetal RHD genotyping with maternal plasma from D- women by use of capillary electrophoresis. STUDY DESIGN AND METHODS: Blood samples were obtained from 8 D+ and 8 D- nonpregnant donors and mixed to make test plasma samples. DNA was extracted and the appropriate conditions relating to the initial sample volume as well as polymerase chain reaction cycle numbers were analyzed to detect the RHD gene with RHD exon 10 primer. Blood samples were also obtained from 13 D- pregnant women ranging from the 12th to 39th weeks of gestation. The presence of the RHD gene and Y-chromosome-specific STR (Y-STR) derived from the fetus was analyzed. The results were compared with the D status of newborns. RESULTS: In samples from 12 D- pregnant women, the RHD gene was detected. In one sample, the RHD gene was not detected but Y-STR loci were demonstrated in this sample, indicating a D- male baby. The results of fetal genotyping were all in concordance with the postpartum samples by serologic tests on D as well as with the sex of newborns. CONCLUSION: Capillary electrophoresis can be used for the determination of fetal RHD status in D- women. This diagnostic method is useful for the noninvasive prenatal diagnosis of the fetal RHD genotyping from D- women. PMID- 18422851 TI - An algorithm based on peripheral CD34+ cells and hemoglobin concentration provides a better optimization of apheresis than the application of a fixed CD34 threshold. AB - BACKGROUND: Optimization of peripheral blood stem cell (PBSC) collection for autologous bone marrow transplantation is necessary for a good standard of care and cost-effectiveness. An algorithm was validated for prediction of the day of maximum peripheral CD34+ cell concentration after mobilization chemotherapy (Day(CD34peak)). STUDY DESIGN AND METHODS: This study compared mobilization and collection variables of a cohort of patients where apheresis was started at the Day(CD34peak) predicted by the algorithm with a patient group where PBSCs were collected when PB CD34+ cell concentration reached 10 per microL per day (Day(CD34threshold)). Day(CD34peak) was calculated according to the equation Day(CD34peak) = -0.41 x Hb(D0) + 0.99 x Day(CD34threshold) + 7.8 (with Hb(D0) representing the hemoglobin value on Day 0). RESULTS: The mean number of apheresis procedures per patient based on the Day(CD34threshold) was 1.74, but decreased to 1.35 when applying the new method (Day(CD34peak)). For lymphomas, the mean number of apheresis procedures decreased from 1.98 to 1.47 (p = 0.03), while in patients with multiple myeloma it did not change significantly (1.23 and 1.26, respectively). Age and primary disease influenced the number of apheresis procedures needed to achieve the collection target. CONCLUSION: The application of our algorithm can lower the number of apheresis procedures by improving the timing, especially in patients suffering from malignant lymphomas with a poor marrow potential after several chemotherapy lines. PMID- 18422852 TI - Herpesvirus prevalence and viral load in healthy blood donors by quantitative real-time polymerase chain reaction. AB - BACKGROUND: After primary infection, human herpesviruses (HHVs) maintain long term latent persistence, often punctuated years later by sporadic episodes of symptomatic lytic activation. Also, blood-borne herpesvirus from healthy persistently infected blood donors can lead to active primary infection of immunocompromised transfusion recipients. STUDY DESIGN AND METHODS: Utilizing a set of newly developed real-time polymerase chain reaction assays for detection and quantification of all eight human herpesviruses, the prevalence and viral DNA load of white cell-enriched blood from 100 randomly selected blood donors from the southeast Texas region are reported. RESULTS: Herpes simplex viruses 1 and 2 (HSV-1 and HSV-2), varicella-zoster virus (VZV), and HHV-8 DNA were not detected in any donor sample. In contrast, Epstein-Barr virus (EBV) (72%) and HHV-7 (65%) were commonly detected, HHV-6 (30%) was often detected (Type B only), and cytomegalovirus (CMV; 1%) was rarely detected. Median viral loads of positive samples (per milliliter of blood) ranged from 4278 for HHV-6 to less than 46 for EBV. CONCLUSIONS: These results suggest that the potential for transfusion mediated transmission of herpesviruses from healthy adult blood donors is high for EBV and HHV-7; moderately high for HHV-6; uncommon for CMV; and rare for HSV 1, HSV-2, VZV, and HHV-8. Perhaps the most remarkable finding in this study was the detection of a single donor sample with greater than 6.1 x 10(7) HHV-6 Type B genome equivalents per mL blood. Given that this extraordinarily high level of HHV-6 DNA was obtained from a healthy adult blood donor, this phenomenon is likely unrelated to active infection or immunodeficiency. PMID- 18422853 TI - Analysis of a high-throughput HLA antibody screening assay for use with platelet donors. AB - BACKGROUND: Passive infusion of HLA antibodies has been implicated in transfusion reactions. A rapid, inexpensive method of screening blood donors for HLA antibodies might reduce the incidence of reactions. A high-throughput microbead flow analyzer HLA antibody detection technique was compared with an enzyme-linked immunosorbent assay (ELISA) method. STUDY DESIGN AND METHODS: Ninety-six apheresis platelet (PLT) donors were tested for antibodies to Class I and II HLA antigens with mixed-antigen microbead-flow analyzer and ELISAs. For both assays, samples reactive in the mixed-antigen assay were tested with a panel-reactive antibody (PRA) assay. Samples reactive in both the mixed-antigen and the PRA assays were considered positive. RESULTS: In the mixed-antigen microbead assay, 46 (48%) samples were reactive to Class I antigens and 20 (21%) to Class II. Further testing in the microbead PRA assay revealed that 34 (35%) had antibodies to Class I antigens, 18 (19%) to Class II, and 42 (44%) to either Class I or Class II. Class I antibodies were present in 56 percent of females and 36 percent of males. In the mixed-antigen ELISA, 4 samples were reactive with Class I antigens, 4 with Class II antigens, and 5 with Class I or Class II. All 5 reactive samples were also reactive in the ELISA PRA assay and were from females. CONCLUSION: The microbead assay was more sensitive than the ELISA and detected antibodies in a large proportion of donors. Samples reactive in the mixed-antigen microbead assay should be confirmed by a second assay before concluding that antibodies are present. PMID- 18422854 TI - Cost-efficient sequence-specific priming-polymerase chain reaction screening for blood donors with rare phenotypes. AB - BACKGROUND: Transfusion support for patients with irregular antibodies to red blood cell (RBC) antigens of high frequency may be hampered by lack of appropriate antigen-negative RBC units. Often, this perceived lack is due to the low number of typed donors. We developed a simple multiplex polymerase chain reaction (PCR) method to screen for donors with rare blood group phenotypes. STUDY DESIGN AND METHODS: A multiplex PCR with sequence-specific priming predicting Yt(a), Co(a), Lu(b), and Kp(b) antigens was developed based on a commercially available system (Extract-N-Amp, Sigma-Aldrich) that obviates the DNA purification step. PCR amplicons were analyzed by size fractionation in a 2 percent agarose gel. Samples representing rare phenotypes were identified by the lack of one of the four visible bands. Donors of blood phenotype O D- ccddee were screened. RESULTS: Excluding the preparation of the reaction mixture and the gel, the whole procedure consisted of five pipetting steps. Hands-on time was 102 minutes for 91 donors. After optimization, interpretable results were obtained in 85 percent of samples without repetition. Among 3422 donors tested, 1 Kp(b-), 6 Co(a-), 10 Yt(a-), and 5 Lu(b-) donors were detected. CONCLUSIONS: Multiplex PCR is a simple, versatile, and cost-efficient method for the screening for donors with rare phenotypes who may be identified by their genotype. If such donor screening is introduced on a broad basis, transfusion support for patients with anti-Co(a), anti-Yt(a), or anti-Lu(b) will be considerably improved. PMID- 18422855 TI - Hemostats, sealants, and adhesives: components of the surgical toolbox. AB - The surgical toolbox is expanding, and newer products are being developed to improve results. Reducing blood loss so that bloodless surgery can be performed may help minimize morbidity and length of stay. As patients, hospital administrators, and government regulators desire less invasive procedures, the surgical technical challenge is increasing. More operations are being performed through minimally invasive incisions with laparoscopic, endoscopic, and robotic approaches. In this setting, tools that can reduce bleeding by causing blood to clot, sealing vessels, or gluing tissues are gaining an increasing importance. Thus, hemostats, sealants, and adhesives are becoming a more important element of surgical practice. This review is designed to facilitate the reader's basic knowledge of these tools so that informed choices are made for controlling bleeding in specific clinical situations. Such information is useful for all members of the operative team. The team includes surgeons, anesthesiologists, residents, and nurses as well as hematologists and other medical specialists who may be involved in the perioperative care of surgical patients. An understanding of these therapeutic options may also be helpful to the transfusion service. In some cases, these materials may be stored in the blood bank, and their appropriate use may reduce demand for other transfusion components. The product classification used in this review includes hemostats as represented by product categories that include mechanical agents, active agents, flowables, and fibrin sealants; sealants as represented by fibrin sealants and polyethylene glycol hydrogels; and adhesives as represented by cyanoacrylates and albumin cross linked with glutaraldehyde. Only those agents approved by the Food and Drug Administration (FDA) and presently available (February 2008) for sale in the United States are discussed in this review. PMID- 18422856 TI - A pilot study for screening blood donors in Taiwan by nucleic acid amplification technology: detecting occult hepatitis B virus infections and closing the serologic window period for hepatitis C virus. AB - BACKGROUND: Blood donors in Taiwan currently are screened for hepatitis B virus (HBV), hepatitis C virus (HCV), and human immunodeficiency virus (HIV) infection by immunoassay. The risk of enzyme immunoassay (EIA)-negative, nucleic acid amplification technology (NAT)-reactive donations is not well understood. This study aimed to screen for such donors in Taiwan by a multiplex test (cobas TaqScreen, Roche) on a commercially available NAT system (cobas s 201 system, Roche). STUDY DESIGN AND METHODS: NAT was performed on donors without prescreening in pools of six and NAT-reactive pools were then resolved to the single donation. Individual-donor NAT-reactive samples were discriminated by a commercially available polymerase chain reaction (PCR)-based diagnostic assay (COBAS AmpliScreen, Roche). Samples with EIA- and NAT-discordant results were investigated with supplemental serologic and confirmatory tests. Each sample taken from follow-up of HBV NAT yield cases was tested for HBV serologic profile, NAT, and viral load. The sensitivity and performance efficacy were also evaluated. RESULTS: The 95 percent limit of detection (LOD) for HBV, HCV, and HIV were 5.09, 11.83, and 62.53 IU per mL, respectively. Among 10,727 seronegative donations, 12 HBV NAT yield cases (0.11%) and 1 HCV NAT yield case (0.01%) were detected. Follow-up results for 1 to 8 months showed that the HCV yield case was a window case and all HBV NAT yield cases were occult carriers. CONCLUSION: The use of NAT detected occult HBV and reduced HCV window period. The yield rate, especially occult HBV, was 10- to 100-fold higher than that in developed, HBV nonendemic countries. Therefore, NAT implementation for routine donor screening in a more cost-effective manner should contribute to safer blood transfusion in Taiwan. PMID- 18422858 TI - Rapid detection of JMH antibodies with recombinant Sema7A (CD108) protein and the particle gel immunoassay. AB - BACKGROUND: At present, identification of antibodies against the high-prevalence JMH antigen is difficult and limited to reference laboratories having panels of rare red blood cell (RBC) specimens in stock. Here, a novel method is described for detection of anti-JMH with particles coated with recombinant semaphorin 7A (Sema7A, CD108), the protein that carries the JMH blood group antigens. STUDY DESIGN AND METHODS: Recombinant Sema7A protein was generated and coupled onto superparamagnetic particles coated with streptavidin. The coated particles were tested in the presence of different serum and plasma samples (11 anti-JMH, 20 other antibodies, and 50 samples from nonimmunized blood donors) with the particle gel immunoassay and flow cytometry. RESULTS: Sema7A-coated particles reacted with all 11 samples containing anti-JMH, but not with samples lacking anti-JMH. In addition, the anti-JMH agglutination scores were higher with Sema7A coated particles than with JMH-positive RBCs in all cases. CONCLUSION: Recombinant blood group proteins have the potential to replace RBCs as antigen carriers for identification of certain RBC alloantibodies. PMID- 18422859 TI - Toxicity related to autologous peripheral blood haematopoietic progenitor cell infusion is associated with number of granulocytes in graft, gender and diagnosis of multiple myeloma. AB - BACKGROUND AND OBJECTIVES: We prospectively evaluated the infusion-related toxicity of autologous peripheral blood progenitor cells (PBPC) in 215 patients with haematologic malignancies or solid tumours. MATERIALS AND METHODS: PBPCs were collected by apheresis after mobilization with chemotherapy and/or granulocyte-colony-stimulating factor (G-CSF). The grafts were cryopreserved in 10% dimethyl sulfoxide (DMSO) and stored in liquid nitrogen. Patients were monitored for vital signs and symptoms of the toxicity during and after infusion. RESULTS: The adverse reactions were reported during 149 (56.9%) infusions. During 21.0% infusions occurred just one symptom classified as grade 1, while during 35.9% occurred multiple symptoms classified as grade 2. Logistic regression analysis showed that female gender, diagnosis of multiple myeloma and number of granulocytes infused per kg body weight were significant predictors of occurrence of adverse reactions during infusion. CONCLUSION: Our results indicate that beside the infused DMSO dose, the composition of graft as well as patient's diagnosis are also very important factors for infusion-related toxicity. PMID- 18422860 TI - Molecular mechanisms of PLD function in membrane traffic. AB - The two mammalian phosphatidylcholine (PC)-selective phospholipase D (PLD) enzymes remove the choline head group from PC to produce phosphatidic acid (PA). PA stimulates phosphatidylinositol(4)phosphate 5-kinases, can function as a binding site for membrane proteins, is required for certain membrane fusion or fission events and is an important precursor for the production of diacylglycerol (DAG). Both PA and DAG are lipids that favor negatively curved membranes rather than planar bilayers and can reduce the energetic barrier to membrane fission and fusion. Recent data provide a mechanistic explanation for the role PLDs play in some aspects of membrane traffic and provide an explanation for why some membrane fusion reactions require PA and some do not. PLDs also act as guanosine triphosphatase-activating proteins for dynamin and may participate with dynamin in the process of vesicle fission. PMID- 18422857 TI - The ongoing variability in blood transfusion practices in cardiac surgery. AB - BACKGROUND: Although blood utilization has been under considerable scrutiny for the past two decades, particularly for surgery, studies comparing perioperative blood transfusion practices between countries are rare, and the evolution of international standards remains unknown. Therefore, the objective of this evaluation was to compare the perioperative transfusion of blood components in cardiac surgery in multiple centers in different countries. STUDY DESIGN AND METHODS: Transfusion practice was investigated prospectively in 70 centers among 16 countries. A total of 5065 randomly selected cardiac surgery patients of the Multicenter Study of Perioperative Ischemia Epidemiology II (EPI II) Study were evaluated. Utilization of red blood cells (RBCs), fresh-frozen plasma (FFP), and platelets (PLTs) was assessed daily, before, during, and after surgery until hospital discharge. RESULTS: Intraoperative RBC transfusion varied from 9 to 100 percent among the 16 countries, and 25 to 87 percent postoperatively (percentage of transfused patients). Similarly, frequency of transfusion of FFP varied from 0 to 98 percent intraoperatively and 3 to 95 percent postoperatively, and PLT transfusion from 0 to 51 and 0 to 39 percent, respectively. Moreover, there were not only marked differences in transfusion rates between centers in different countries but also in interinstitutional comparison of multiple centers within countries. CONCLUSION: In cardiac surgical patients, marked variability in transfusion practice exists between centers in various countries and suggests differences in perioperative practice patterns as well as possible inappropriate use. International standardization of perioperative practice patterns as well as transfusion regimes appears necessary. PMID- 18422862 TI - Quantification of sexual steroid hormones in faeces of Iberian wolf (Canis lupus signatus): a non-invasive sex typing method. AB - The determination of gender in wild animals is essential for behavioural and ecological studies, and also for conservation. The objectives of this study were (i) the determination of gender in faecal samples of Iberian wolf based on the differential concentrations of sexual steroid hormones (SSH) and (ii) to analyse the profiles of SSH in males and females (considering the gender determination carried out previously) during the non-reproductive and reproductive periods. The quantification of androgens (testosterone, T), progestin (progesterone, P) and oestrogen (oestradiol, E) was conducted by means of enzyme immunoassay. The k means conglomerate analysis showed that the 59 faecal samples grouped into three different conglomerates, considering SSH levels. Groups 1 and 2 showed higher levels of T than group 3. Therefore, the faecal samples included in groups 1 and 2 (17 samples) corresponded to males and those of group 3 (42 samples) to females. The levels of T + P + E and T/P were higher in the group of males than in the group of females. The results of this study also showed that levels of T in males were higher during the reproductive period than in the non-reproductive period. However, the concentrations of P and E turned out to be higher during the non-reproductive season. In females, the levels of the three hormones (T, P and E) were higher during the reproductive period. PMID- 18422861 TI - Relationship between low ultraviolet B irradiance and higher breast cancer risk in 107 countries. AB - Epidemiological data show an inverse relationship between vitamin D levels and breast cancer incidence. This study investigates the relationship of modeled and measured serum 25-hydroxyvitamin D [25(OH)D] levels with age-standardized incidence rates of breast cancer in 107 countries. The hypothesis being tested is that breast cancer incidence is inversely related to geographically-dependent cutaneous sunlight exposure. A multiple regression approach was used to examine the contributions of ultraviolet B (UVB) irradiance to age-standardized incidence rates of breast cancer in the 107 countries with data on these covariates-total column ozone thickness, per capita intake of alcohol and energy from animal and vegetable sources, cigarettes, proportion of female population overweight, and total fertility. Age-standardized incidence rates were substantially higher at latitudes distant from the equator (R2 = 0.43, p < 0.0001). The dose-response gradient between modeled serum 25(OH)D levels and incidence rates of breast cancer followed a standard inverse dose-response curve. Increasing increments in serum 25(OH)D in the range above 22 ng/mL were associated with incrementally lower incidence rates of breast cancer. According to multiple regression, UVB irradiance adjusted for cloud cover was inversely associated with incidence rates (p = 0.04) after controlling for covariates. Intake of energy from animal sources was also positively associated with incidence rates (p < 0.01). The overall coefficient of determination, R2, was 0.81 (p < 0.0001). There was a protective effect of UVB irradiance on risk of breast cancer that was independent of fertility rate, proportion of the population overweight, alcohol intake, animal energy intake, and other covariates. PMID- 18422863 TI - Dynamics of follicular fluid in one-humped camel (Camelus dromedarius). AB - In the present study, ovarian follicular fluid and serum biochemical, hormonal, electrolytes and amino acids profiles in female dromedary camel (Camelus dromedarius), were investigated. Fluid from small (2-6 mm) and large follicles (7 20 mm) and blood samples were collected from 25 clinically healthy adult female camels. The concentrations of glucose, cholesterol, triglycerides, high-density lipoproteins, urea, total proteins, albumin, globulin, fibrinogen, alanine aminotransferase, aspartate aminotransferase and tri-iodothyronine were lower (p < or = 0.05) in large follicles when compared with the small follicles. However, the concentrations of low-density lipoproteins, uric acid, creatinine, alkaline phosphatase and acid phosphatase in small and large follicles did not differ. The concentrations of oestradiol 17-beta and progesterone were higher (p < or = 0.05) in large follicles. The serum concentrations of these hormones were many folds lower (p < or = 0.05) than those of follicular fluid. Among electrolytes, the concentration of phosphorus was higher (p < or = 0.05) in the large follicles, while that of potassium and chloride were lower (p < or = 0.05) in the small follicles. Serum concentrations of sodium, chloride, calcium and phosphorous were higher (p < or = 0.05), while that of potassium lower (p < or = 0.05) than corresponding concentrations in the follicular fluid. The concentrations of leucine and arginine were higher (p < or = 0.05) in follicular fluid when compared with serum concentrations, while the reverse was true for other amino acids. In conclusion, this study is indicative of either low or high concentrations of certain biochemical metabolites, hormones, electrolytes and amino acids in small and large follicles for the individual roles that they play in the growth and development of follicles in the one-humped she-camel. PMID- 18422864 TI - A study of the complications of small bore 'Seldinger' intercostal chest drains. AB - BACKGROUND AND OBJECTIVE: Use of small bore chest drains (<14F), inserted via the Seldinger technique, has increased globally over the last few years. They are now used as first line interventions in most acute medical situations when thoracostomy is required. Limited data are available on the associated complications. In this study, the frequency of complications associated with 12F chest drains, inserted using the Seldinger technique, was quantified. METHODS: A retrospective case note audit was performed of consecutive patients requiring pleural drainage over a 12-month period. One hundred consecutive small bore Seldinger (12F) chest drain insertions were evaluated. RESULTS: Few serious complications occurred. However, 21% of the chest drains were displaced ('fell out') and 9% of the drains became blocked. This contributed to high morbidity rates, with 13% of patients requiring repeat pleural procedures. The frequency of drain blockage in pleural effusion was reduced by administration of regular normal saline drain flushes (odds ratio for blockage in flushed drains compared with non-flushed drains 0.04, 95% CI: 0.01-0.37, P < 0.001). CONCLUSIONS: Regular chest drain flushes are advocated in order to reduce rates of drain blockage, and further studies are needed to determine optimal fixation strategies that may reduce associated patient morbidity. PMID- 18422865 TI - Arterial blood lactate is a useful guide to when rehabilitation should be instigated in COPD. AB - BACKGROUND AND OBJECTIVE: Systemic effects of COPD include skeletal muscle dysfunction; the lactate threshold (LT) is an index of such dysfunction. However, it is not feasible in daily clinical practice to accurately determine the LT in all patients with COPD. There is no simple, practical and non-invasive index for determining the time at which rehabilitation should start. Previous studies have shown that the LT corresponds to the point at which the blood lactate concentration is 0.5 mmol/L above baseline (LTDelta0.5 mmol/L). The aim of the study was to assess the value of LTDelta0.5 mmol/L as an index of selecting candidates for pulmonary rehabilitation in COPD patients. METHODS: Eighteen male outpatients with COPD were enrolled. Exercise tolerance based on the 12-min walk test and treadmill exercise test, lactate concentrations and activities of daily living before and after individual exercise stress testing were assessed. RESULTS: There were no significant differences in the 12-min walk distance or metabolic equivalents between patients with and without a 0.5 mmol/L or more increase in lactate from baseline to post-walk test. However, significant differences were observed in activities of daily living, as assessed using Fletcher's scale, between those with and without a 0.5 mmol/L or greater increase in lactate from baseline to post-walk test. CONCLUSIONS: In daily clinical practice, a 0.5 mmol/L or greater increase in lactate from baseline to post-12 min walk test may be utilized as an index to determine when COPD patients should start rehabilitation, or to assess muscle alterations in the lower extremities in COPD patients undergoing rehabilitation. PMID- 18422866 TI - Spontaneous regression of small cell lung cancer. AB - Spontaneous regression of cancers is extremely rare and is associated with specific malignancies. Spontaneous regression of bronchogenic lung cancer has rarely been reported, and regression of small cell lung cancer is even less common. Such regression is generally ascribed to immunological factors but is not well understood. This case report describes a patient with spontaneous regression of small cell lung cancer that has persisted for 11 years and considers possible mechanisms. PMID- 18422867 TI - Procalcitonin is a valuable prognostic marker in ARDS caused by community acquired pneumonia. AB - BACKGROUND AND OBJECTIVE: ARDS is life-threatening acute respiratory failure, and pneumonia is one of the most common causes of direct ARDS. Procalcitonin (PCT) has been evaluated for its utility in determining the aetiology of community acquired pneumonia (CAP), choice of antibiotics and prediction of outcome. This study evaluated the role of PCT in predicting the outcome of patients with ARDS caused by severe CAP. METHODS: This was a prospective observational study conducted from September 2002 to December 2003. The plasma PCT was analysed at baseline, 24 and 72 h after enrolment and measured by ELISA. RESULTS: Of the 22 patients with ARDS caused by CAP and enrolled in the study, 17 (77.3%) were alive 14 days after admission and five (22.7%) had died. The survivors had lower APACHE II scores (22.2 +/- 4.6 vs 30.6 +/- 9.6, P = 0.031), pneumonia severity index (141.9 +/- 2.2 vs 195.6 +/- 23.8, P = 0.005) and lower plasma PCT at baseline (9.83 +/- 3.54 vs 106.70 +/- 67.86, P = 0.004), at 24 h (10.51 +/- 5.39 vs 81.32 +/- 57.68, P = 0.014) and at 72 h (2.03 +/- 0.76 vs 19.57 +/- 6.67, P = 0.005). CONCLUSION: PCT analysed within 72 h of the onset of ARDS predicted mortality of patients with ARDS caused by severe CAP. PMID- 18422868 TI - Assessing adherence and factors associated with adherence in young children with asthma. AB - BACKGROUND AND OBJECTIVE: Adherence with preventive asthma medication by young children is an important factor when evaluating a suboptimal response to treatment. However, few data exist regarding the accuracy of subjective measures of adherence and factors associated with adherence in young children. METHODS: Fifty-one asthmatic children aged 18 months to 7 years had their use of preventive asthma medication monitored using an electronic monitoring device (Smartinhaler) for 1 month. At a follow-up visit the child's parent was asked how often medication had been given and they also completed a confidential questionnaire that included questions about medication usage, barriers to optimal adherence and parenting. The treating physician made an estimate of the child's likely use of medication. RESULTS: The median use of medication as determined by the Smartinhaler was 70.5% (range 21.4-100%). The parents' verbal reports (85.1%) and questionnaire responses (84.2%) overestimated medication usage. The physician was not able to determine which parents correctly estimated their child's use of medication (P = 0.28). The child's age, level of parental education and annual family income did not influence adherence. Parents reported simply 'forgetting' or their child's 'reaction to being given medication' as the principal barriers to adherence. There was a significant association between how stressful the parent found parenting and adherence (P = 0.05). CONCLUSION: Adherence with preventive medication, even within the context of a research study, was generally low and highly variable. Subjective measures of adherence were found to overestimate adherence in young asthmatics. PMID- 18422869 TI - Diagnostic value of carcinoembryonic antigen in malignant pleural effusion: a meta-analysis. AB - BACKGROUND AND OBJECTIVE: Conventional tests are not always helpful in making a diagnosis of malignant pleural effusion (MPE). Many studies have investigated the utility of pleural carcinoembryonic antigen (CEA) in the early diagnosis of MPE. The present meta-analysis determined the accuracy of CEA measurement in the diagnosis of MPE. METHODS: A systematic review of English language studies was conducted and data on the accuracy of pleural CEA concentrations in the diagnosis of MPE were pooled using random effects models. Receiver operating characteristic curves were used to summarize the overall test performance. RESULTS: Forty-five studies met the inclusion criteria for the meta-analysis. The summary estimates for CEA in the diagnosis of MPE were: sensitivity 0.54 (95% CI: 0.52-0.55), specificity 0.94 (95% CI: 0.93-0.95), positive likelihood ratio 9.52 (95% CI: 6.97-13.01), negative likelihood ratio 0.49 (95% CI: 0.44-0.54) and diagnostic odds ratio 22.5 (95% CI: 15.6-32.5). Analysis of a subset of 11 studies which examined the value of pleural CEA in ruling out a diagnosis of malignant mesothelioma found that the sensitivity and specificity of a CEA level exceeding cut-off values were 0.97 (95% CI: 0.93-0.99) and 0.60 (95% CI: 0.55-0.65), respectively. CONCLUSIONS: Measurement of pleural CEA is likely to be a useful diagnostic tool for confirming MPE, and is also helpful in the differential diagnosis between malignant pleural mesothelioma and metastatic lung cancer. The results of CEA assays should be interpreted in parallel with clinical findings and the results of conventional tests. PMID- 18422870 TI - Functional analysis and expression characteristics of chloroplastic Prx IIE. AB - Peroxiredoxins (Prxs) are ubiquitous thiol-dependent peroxidases capable of eliminating a variety of peroxides through reactive catalytic cysteines, which are regenerated by reducing systems. Based on amino acid sequences and their mode of catalysis, five groups of thiol peroxidases have been distinguished in plants, and type II Prx is one of them with representatives in many sub-cellular compartments. The mature form of poplar chloroplastic Prx IIE was expressed as a recombinant protein in Escherichia coli. The protein is able to reduce H2O2 and tert-butyl hydroperoxide and is regenerated by both glutaredoxin (Grx) and thioredoxin (Trx) systems. Nevertheless, compared with Trxs, Grxs, and more especially chloroplastic Grx S12, are far more efficient reductants towards Prx IIE. The expression of Prx IIE at both the mRNA and protein levels as a function of organ type and abiotic stress conditions was investigated. Western blot analysis revealed that Prx IIE gene is constitutively expressed in Arabidopsis thaliana, mostly in young and mature leaves and in flowers. Under photo-oxidative treatment and water deficit, almost no change was observed in the abundance of Prx IIE in A. thaliana, while the level of Prx Q (one of the two other chloroplastic Prxs with 2-Cys Prx) increased in response to both stresses, indicating that plastidic members of the Prx family exhibit specific patterns of expression under stress. PMID- 18422872 TI - Haemonchus contortus calreticulin binds to C-reactive protein of its host, a novel survival strategy of the parasite. AB - Calreticulin (CalR), a Ca(2+) binding multifunctional protein, is secreted by the parasitic nematode Haemonchus contortus. We have earlier observed binding of this protein to a 24-kDa polypeptide (p24) present in an enriched preparation of prothrombin. In the present study, the identity of p24 was established as a C reactive protein (CRP) by several criteria. CalR binding to CRP is an elegant strategy devised by the parasite to survive in the host. The secreted CalR may achieve this either by limiting the free concentration of CRP, which has antiparasite activity or inhibit the activation of the classical complement pathway triggered on binding of CRP to C1q protein. CalR binding to CRP would also ensure a check on the procoagulant activity of the CRP enabling parasite to feed on the host blood. Thus, targeting CalR could be a novel strategy to tackle this parasite, which has developed resistance to many anthelmintics. PMID- 18422873 TI - Activity switches of rhodopsin. AB - Rhodopsin, the visual pigment of the rod photoreceptor cell contains as its light sensitive cofactor 11-cis retinal, which is bound by a protonated Schiff base between its aldehyde group and the Lys296 side chain of the apoprotein. Light activation is achieved by 11-cis to all-trans isomerization and subsequent thermal relaxation into the active, G protein-binding metarhodopsin II state. Metarhodopsin II decays via two parallel pathways, which both involve hydrolysis of the Schiff base eventually to opsin and released all-trans retinal. Subsequently, rhodopsin's dark state is regenerated by a complicated retinal metabolism, termed the retinoid cycle. Unlike other retinal proteins, such as bacteriorhodopsin, this regeneration cycle cannot be short cut by light, because blue illumination of active metarhodopsin II does not lead back to the ground state but to the formation of largely inactive metarhodopsin III. In this review, mechanistic details of activating and deactivating pathways of rhodopsin, particularly concerning the roles of the retinal, are compared. Based on static and time-resolved UV/Vis and FTIR spectroscopic data, we discuss a model of the light-induced deactivation. We describe properties and photoreactions of metarhodopsin III and suggest potential roles of this intermediate for vision. PMID- 18422871 TI - Effect of GPI anchor moiety on the immunogenicity of DNA plasmids encoding the 19 kDa C-terminal portion of Plasmodium falciparum MSP-1. AB - The glycosylphosphatidylinositol (GPI)-anchored Plasmodium falciparum merozoite surface protein 1 (MSP-1) is a widely studied malaria vaccine candidate. The C terminal 19-kDa portion of MSP-1 (MSP-1(19)) is of particular interest because this polypeptide moiety remains bound to the parasite even after erythrocyte invasion, while the remainder of MSP-1 is shed during invasion. Studies have shown that antibodies against MSP-1(19) inhibit merozoite invasion of erythrocytes efficiently, and that MSP-1(19) produces protective immunity in mice and monkeys. To investigate the efficacy of MSP-1(19 )DNA vaccine and role of GPI anchor moiety in the immunogenicity of MSP-1(19), we constructed expression vectors that produce MSP-1(19) as either secretory or GPI-anchored polypeptide. Both constructs efficiently expressed MSP-1(19) in transfected HEK-293 cells. While the recombinant plasmid lacking GPI anchor signal sequence expressed MSP 1(19) mainly as secreted polypeptide, that containing GPI anchor signal sequence produced GPI-anchored MSP-1(19 )on cell surface. In immunized mice, both constructs produced substantial levels of MSP-1(19)-specific IgG1, IgG2a, IgG2b, IgG3, IgA and IgM antibodies. In both cases, the IgG1 level was significantly higher than other isotypes. Interestingly, the plasmid containing GPI anchor signal sequence produced significantly higher levels of IgG2a and IgG2b than the plasmid that lacks GPI signal sequence. PMID- 18422874 TI - Photoprotection by porcine eumelanin against singlet oxygen production. AB - Melanin, a major pigment found in retinal pigment epithelium (RPE) cells, is considered to function in dual roles, one protective and one destructive. By quenching free radical species and reactive oxygen species (ROS) melanin counteracts harmful redox stress. However, melanin is also thought to be capable of creating ROS. In this destructive role, melanin increases redox strain in the cell. This study uses readily available eumelanin extracted from porcine RPE cells as a more authentic model than synthetic melanin to determine specific mechanisms of melanin activity with regard to singlet oxygen in the presence and absence of rose bengal, a singlet-oxygen photosensitizer. Optical detection of singlet-oxygen was determined by monitoring the bleaching of p nitrosodimethylaniline in the presence of histidine. Production of singlet oxygen in aqueous oxygen-saturated solutions of rose bengal without eumelanin was readily accomplished. In contrast, detection of singlet oxygen in oxygen saturated solutions of eumelanin without rose bengal failed, consistent with results of others. However, a significant decrease in singlet oxygen production by rose bengal was observed in the presence of eumelanin. After correction for light absorption and chemical bleaching of eumelanin, the results show that eumelanin also provides a photoprotective mode arising from chemistry, that is, not just the physical process of light absorption followed by energy dissipation as heat. PMID- 18422875 TI - In vivo measurement of vascular modulation in experimental tumors using a fluorescent contrast agent. AB - We compared the effectiveness of three optical techniques based on fluorescence imaging and spectroscopy with indocyanine green (ICG) contrast agent to evaluate in vivo the disruption of the active vasculature induced by a vascular targeting agent. The blood perfusion of the MDA-MB-435 tumor model transplanted in nude mice was estimated from the signal of the contrast agent measured immediately after its systemic injection in mice. Optical measurements were performed using a fluorescence imaging setup and a fiber-based time correlated single photon counting (TCSPC) apparatus. This latter apparatus was used to measure the tumor fluorescence in transmittance geometry and the change in the basal optical absorption induced by the contrast agent, thus providing an alternative estimation of the blood content in the tumor. Mice were divided into four groups. Three groups were treated with different doses of the vascular disrupting agent ZD6126, the fourth group (control group) received the drug vehicle only. Optical measurements were carried out 3 h after pharmacologic treatment. After 24 h, mice were killed, tumors were excised and the extent of necrosis was evaluated with standard histologic analysis. On fluorescence imaging ICG emission from tumors of mice treated with ZD6126 significantly was lower compared with the emission from control mice. The histologic sections also showed a significantly higher amount of necrosis in tumors of treated mice. Both these findings, which correlate with each other, indicate an effective vascular shutdown induced by the drug. However, ICG fluorescence measured with the TCSPC apparatus in transmittance geometry and the estimate of the change in optical absorption did not allow a statistically significant differentiation between treated and control groups. PMID- 18422876 TI - Influence of pH upon surface-enhanced enzyme-catalyzed luminol chemiluminescence at vicinity of nanoscale-corrugated gold and silver films. AB - Au and Ag biochips were fabricated to investigate the influence of pH upon the chemiluminescence (CL) of luminol at vicinity of surface-adsorbed peroxidase. A nanoscaled-corrugation of the metal induces an enhancement of the luminol CL which is maximal in the pH range favoring peroxidase catalysis and greater for gold than for silver. This is the proof that, in the CL process, the reactions involving peroxidase are surface-enhanced near corrugated surfaces. PMID- 18422877 TI - Kinetic properties of hexameric tyrosinase from the crustacean Palinurus elephas. AB - Tyrosinases catalyze hydroxylation of monophenols to o-diphenols and their subsequent oxidation to o-quinones, whereas catecholoxidases catalyze only the latter reaction. Both enzymes occur in all organisms and are Type 3 copper proteins that perform the first steps of melanin formation. In arthropods, they play an essential role in the sclerotization of the exoskeleton. Very few phenoloxidases are characterized structurally or kinetically and the existence of an actual tyrosinase activity has not been demonstrated in most cases. Here we present for the first time a complete kinetic characterization of a tyrosinase from a crustacean (Palinurus elephas) including the influence of inhibitors. In contrast to most tyrosinases which are monomeric or dimeric, this tyrosinase occurs as a hexamer. However, the data did not indicate cooperativity in steady state kinetics for the two substrates used, the monophenol tyramine and the diphenol dopamine. Mimosine as well as phenylthiourea (PTU) inhibited both monophenolhydroxylase and diphenoloxidase activity. Inhibition by mimosine was competitive, whereas PTU was a noncompetitive inhibitor. Furthermore, for the diphenolase activity substrate inhibition was observed, which was apparently abolished by adding PTU. These observations lead to the hypothesis that a secondary, allosteric binding site exists, which binds dopamine and PTU and reduces the catalytic activity. PMID- 18422878 TI - Ultra-low temperature oxidation of 5,6-dihydroxyindole: a novel approach to study synthetic melanogenesis. AB - The detailed structure of melanin remains elusive due to the complexity and insolubility of the pigment. Herein we describe a novel oxidation of 5,6 dihydroxyindole (DHI) as a means to characterize soluble intermediates formed prior to oligomerization. The approach entails the use of a biomimetic copper peroxo oxidant, at ultra-low temperature (-78 degrees C). DHI oxidized by [LCuII(O2)CuIIL]B(C6F5)4 (L=2,6,10-trimethyl-2,6,10-triazaundecane) under argon produces the one electron oxidation product, semiquinone radical, which is spectroscopically observed at -78 degrees C. MS analysis of the reaction mixture reveals the DHI dimer as well as other extensively oxidized DHI units. PMID- 18422879 TI - Functional expression, targeting and Ca2+ signaling of a mouse melanopsin-eYFP fusion protein in a retinal pigment epithelium cell line. AB - Melanopsin, first discovered in Xenopus melanophores, is now established as a functional sensory photopigment of the intrinsically photosensitive retinal ganglion cells. These ganglion cells drive circadian rhythm and pupillary adjustments through projection to the brain. Melanopsin shares structural similarities with all known opsins. Comprehensive characterization of melanopsin with respect to its spectral properties, photochemical cascade and signaling partners requires a suitable recombinant system and high expression levels. This combination has not yet been described. To address this issue, we have expressed recombinant mouse melanopsin in several cell lines. Using enhanced yellow fluorescent protein (eYFP) as a visualization tag, expression was observed in all cell lines. Confocal microscopy revealed that melanopsin was properly routed to the plasma membrane only in retinal pigment epithelium (RPE)-derived D407 cells and in human embryonic kidney (HEK) cells. Further, we performed intracellular calcium measurements in order to probe the melanopsin signaling activity of this fusion protein. Transfected cells were loaded with the calcium indicator Fura2 AM. Upon illumination, an immediate but transient calcium response was observed in HEK as well as in D407 cells, while mock-transfected cells showed no calcium response under identical conditions. Supplementation with 11-cis retinal or all trans retinal enhanced the response. After prolonged illumination the cells became desensitized. Thus, RPE-derived cells expressing recombinant melanopsin may constitute a suitable system for the study of the structural and functional characteristics of melanopsin. PMID- 18422880 TI - Singlet oxygen reacts with 2',7'-dichlorodihydrofluorescein and contributes to the formation of 2',7'-dichlorofluorescein. AB - There are controversial reports in the literature concerning the reactivity of singlet oxygen ((1)O(2)) with the redox probe 2',7'-dichlorodihydrofluorescein (DCFH). By carefully preparing solutions in which (1)O(2) is quantitatively generated in the presence of DCFH, we were able to show that the formation rate of the fluorescent molecule derived from DCFH oxidation, which is 2',7' dichlorofluorescein (DCF), increases in D(2)O and decreases in sodium azide, proving the direct role of (1)O(2) in this process. We have also prepared solutions in which either (1)O(2) or dication (MB(2+)) and semi-reduced (MB) radicals of the sensitizer and subsequently super-oxide radical (O(2)(-)) are generated. The absence of any effect of SOD and catalase ruled out the DCFH oxidation by O(2)(-), indicating that both (1)O(2) and MB(2+) react with DCFH. Although the formation of DCF was 1 order of magnitude larger in the presence of MB(2+) than in the presence of (1)O(2), considering the rate of spontaneous decays of these species in aqueous solution, we were able to conclude that the reactivity of (1)O(2) with DCFH is actually larger than that of MB(2+). We conclude that DCFH can continue to be used as a probe to monitor general redox misbalance induced in biologic systems by oxidizing radicals and (1)O(2). PMID- 18422881 TI - Presence of rhodopsin and porphyropsin in the eyes of 164 fishes, representing marine, diadromous, coastal and freshwater species--a qualitative and comparative study. AB - There are two types of visual pigments in fish eyes; most marine fishes have rhodopsin, while most freshwater fishes have porphyropsin. The biochemical basis for this dichotomy is the nature of the chromophores, retinal (A1) and 3 dehydroretinal (A2), each of which is bound by an opsin. In order to study the regional distribution of these visual pigments, we performed a new survey of the visual pigment chromophores in the eyes of many species of fish. Fish eyes from 164 species were used to examine their chromophores by high-performance liquid chromatography--44 species of freshwater fish, 20 of peripheral freshwater fish (coastal species), 10 of diadromous fish and 90 of seawater fish (marine species) were studied. The eyes of freshwater fish, limb freshwater fish and diadromous fish had both A1 and A2 chromophores, whereas those of marine fish possessed only A1 chromophores. Our results are similar to those of previous studies; however, we made a new finding that fish which live in freshwater possessed A1 if living near the sea and A2 if living far from the sea if they possessed only one type of chromophore. PMID- 18422882 TI - Wet forced-air warming blankets are ineffective at maintaining normothermia. AB - BACKGROUND: Forced-air warming systems have proven effective in preventing perioperative hypothermia. To date, reported adverse events relate primarily to overheating and thermal injuries. This study uses a simple model to show that forced-air warming blankets become ineffective if they get wet. METHODS: Temperature sensor probes were inserted into three 1-liter fluid bags. Group C bags served as the control. Groups D (dry) and W (wet) bags were placed on Bair Hugger(R) Model 555 (Arizant Healthcare, Inc., Eden Prairie, MN, USA) pediatric underbody blankets. The warming blanket for Group W bags was subsequently wet with irrigation fluid. Temperature was documented every 5 min. This model was repeated two times for a total of three cycles. Statistical analysis was performed using anova for repeated measures. RESULTS: Starting temperatures for each model were within a 0.3 degrees C range. Group C demonstrated a steady decline in temperature. Group D maintained and slightly increased in temperature during the observation period, while Group W exhibited a decrease in temperature at a rate similar to Group C. These results were significant at P < 0.005. CONCLUSIONS: A wet forced-air warming blanket is ineffective at maintaining normothermia. Once wet, the warming blanket resulted in cooling similar to the control group. PMID- 18422883 TI - An unusual complication following laparoscopic pyloromyotomy and a classic pediatric anesthesia dilemma. PMID- 18422884 TI - Intraoperative alertness of a rise in serum potassium by double counting of the heart rate during blood transfusion in an infant. PMID- 18422885 TI - Pediatric video laryngoscope rescue for a difficult neonatal intubation. PMID- 18422886 TI - Human papillomavirus L1 protein expressed in tobacco chloroplasts self-assembles into virus-like particles that are highly immunogenic. AB - Cervical cancer is the second most prevalent cancer in women worldwide. It is linked to infection with human papillomavirus (HPV). As the virus cannot be propagated in culture, vaccines based on virus-like particles have been developed and recently marketed. However, their high costs constitute an important drawback for widespread use in developing countries, where the incidence of cervical cancer is highest. In a search for alternative production systems, the major structural protein of the HPV-16 capsid, L1, was expressed in tobacco chloroplasts. A very high yield of production was achieved in mature plants (approximately 3 mg L1/g fresh weight; equivalent to 24% of total soluble protein). This is the highest expression level of HPV L1 protein reported in plants. A single mature plant synthesized approximately 240 mg of L1. The chloroplast-derived L1 protein displayed conformation-specific epitopes and assembled into virus-like particles, visible by transmission electron microscopy. Furthermore, leaf protein extracts from L1 transgenic plants were highly immunogenic in mice after intraperitoneal injection, and neutralizing antibodies were detected. Taken together, these results predict a promising future for the development of a plant-based vaccine against HPV. PMID- 18422887 TI - Spatial and temporal expression of endosperm transfer cell-specific promoters in transgenic rice and barley. AB - Two putative endosperm-specific rice genes, OsPR602 and OsPR9a, were identified from database searches. The promoter regions of these genes were isolated, and transcriptional promoter:beta-glucuronidase (GUS) fusion constructs were stably transformed into rice and barley. The GUS expression patterns revealed that these promoters were active in early grain development in both rice and barley, and showed strongest expression in endosperm transfer cells during the early stages of grain filling. The GUS expression was similar in both rice and barley, but, in barley, expression was exclusively in the endosperm transfer cells and differed in timing of activation relative to rice. In rice, both promoters showed activity not only in the endosperm transfer cells, but also in the transfer cells of maternal tissue and in several floral tissues shortly before pollination. The expression patterns of OsPR602 and OsPR9a in flowers differed. The similarity of expression in both rice and barley suggests that these promoters may be useful to control transgene expression in the transfer cells of cereal grains with the aim of altering nutrient uptake or enhancing the barrier against pathogens at the boundary between maternal tissue and the developing endosperm. However, the expression during floral development should be considered if the promoters are used in rice. PMID- 18422888 TI - TILLMore, a resource for the discovery of chemically induced mutants in barley. AB - A sodium azide-mutagenized population of barley (cv. 'Morex') was developed and utilized to identify mutants at target genes using the 'targeting induced local lesions in genomes' (TILLING) procedure. Screening for mutations at four agronomically important genes (HvCO1, Rpg1, eIF4E and NR) identified a total of 22 new mutant alleles, equivalent to the extrapolated rate of one mutation every 374 kb. All mutations except one were G/C to A/T transitions and several (approximately 68%) implied a change in protein amino acid sequence and therefore a possible effect on phenotype. The high rate of mutation detected through TILLING is in keeping with the high frequency (32.7%) of variant phenotypes observed amongst the M(3) families. Our results indicate the feasibility of using this resource for both reverse and forward genetics approaches to investigate gene function in barley and related crops. PMID- 18422889 TI - Precision breeding for novel starch variants in potato. AB - Potato can be used as a source of modified starches for culinary and industrial processes, but its allelic diversity and tetraploid genome make the identification of novel alleles a challenge, and breeding such alleles into elite lines is a slow and difficult process. An efficient and reliable strategy has been developed for the rapid introduction and identification of new alleles in elite potato breeding lines, based on the ethylmethanesulphonate mutagenesis of dihaploid seeds. Using the granule-bound starch synthase I gene (waxy) as a model, a series of point mutations that potentially affect gene expression or enzyme function was identified. The most promising loss-of-function allele (waxy(E1100)) carried a mutation in the 5'-splice donor site of intron 1 that caused mis-splicing and protein truncation. This was used to establish elite breeding lineages lacking granule-bound starch synthase I protein activity and producing high-amylopectin starch. This is the first report of rapid and efficient mutation analysis in potato, a genetically complex and vegetatively propagated crop. PMID- 18422890 TI - Antineuronal antibodies and electroneurophysiological studies in pediatric patients with neuropsychiatric systemic lupus erythematosus. AB - Neuropsychiatric systemic lupus erythematosus (NPSLE) is one of the most difficult manifestations of lupus to diagnose. Measurement of serum brain autoantibodies and assessment of cognitive function by electroneurophysiological studies (electroencephalogram (EEG) and P300) have contributed to an earlier and a more specific diagnosis of NPSLE. Thus, we were stimulated to assess the value of serum antineuronal antibodies and electroneurophysiological studies in diagnosis and early prediction of NPSLE. To investigate this, assessment of serum antineuronal antibodies and cognitive function (clinically and by electroneurophysiological studies) was done in 30 lupus patients [14 (46.7%) with and 16 (53.3%) without clinical evidence of NPSLE] in comparison with 30 healthy matched subjects. Patients without clinical evidence of NPSLE were followed-up clinically by monthly neuropsychiatric evaluation for 18 months. Seropositivity for antineuronal antibodies and abnormalities of EEG and P300 (prolonged latency and/or low amplitude) were found in 60%, 50% and 70%, respectively of lupus patients. During follow-up, 8 out of the 16 patients without clinical evidence of NPSLE developed such evidence [six (75%) had antineuronal seropositivity, five (62.5%) had abnormal EEG, six (75%) had P300 abnormalities and all had at least one abnormal result of these parameters at the time of initial evaluation before clinical presentation of NPSLE]. In conclusion, serum antineuronal antibodies and electroneurophysiological studies may be reliable parameters for diagnosis and early prediction of NPSLE, especially when combined together, before clinical manifestations ensue. Further studies on a large scale are warranted to evaluate the predictive value of these parameters in NPSLE. PMID- 18422891 TI - Clinical assessment of nasal decongestion test by VAS in adolescents. AB - Nasal airflow, as measured by rhinomanometry, is frequently impaired in allergic rhinitis (AR). The decongestion test evaluates whether the application of an intranasal vasoconstrictor drug increases nasal airflow. The aim of this study was to verify the suitability of the use of the visual analogue scales (VAS) as a surrogate for rhinomanometry in the decongestion test assessment in adolescents with atopic rhinitis. Forty adolescents [16 males and 24 females, mean age 15 (s.d. 2) yr] with AR were studied. Nasal symptoms, VAS, rhinomanometry, and nasal decongestion test were assessed in all patients. A significant association was observed between VAS and nasal airflow after performing the decongestion test (Spearman's r is -51.7%, p < 0.001). The associated sensitivity and specificity were 84.8 (95% confidence interval, CI 68.1-94.8) and 85.7 (95% CI 42.2-97.6), respectively. The corresponding area under the receiver operating characteristic (ROC) curve of 0.83 (95% CI 0.67-0.93) indicated a good discriminating ability for the decongestion measured on the VAS scale. In conclusion, the use of VAS appears as clinically relevant, in that it allows, with a fair reliability, to perform the decongestion test in the absence of rhinomanometry. PMID- 18422892 TI - IgE antibody responses in young children with atopic dermatitis. AB - In 2184 young children aged 13-24 months with atopic dermatitis (SCORAD 5-59) serum IgE antibodies to a standard panel of food and inhalant allergens were assayed. The frequency of positive IgE responses (>0.35 kU/l) increased with greater severity of skin disease. A significant minority of infants had levels of IgE antibody to foods to suggest they were at risk of acute reaction to those foods (7% to hen's egg, 3% to cow's milk, 4% to peanut). Our findings indicate that the frequency of positive IgE responses is related to disease severity and suggest that differences in the time course of the development of IgE responses to food, which are at maximum prevalence within the first year of life, while inhalant allergies, are still developing between 1 and 2 yr and beyond. PMID- 18422893 TI - Transcriptome analysis of Arabidopsis roots treated with signaling compounds: a focus on signal transduction, metabolic regulation and secretion. AB - Gene expression in response to signaling molecules has been well studied in the leaves of the model plant species Arabidopsis thaliana. However, knowledge of gene expression and metabolic regulation at the root level is limited. Here, the signaling compounds salicylic acid (SA), methyl jasmonate (MeJA) and nitric oxide (NO) were applied exogenously to induce various defense responses in roots, and their effect was studied using a combination of genomic, molecular and biochemical approaches. Genes involved in defense signaling/activation, cellular redox state, metabolism, transcription factors and membrane transport were altered in expression following treatment with SA, MeJA and NO. In addition, it was found that SA-, MeJA- and NO-elicited roots increased the root exudation of phytochemicals compared with the roots of nontreated control plants. Transport systems likely to be involved in the root exudation of phytochemicals, including the MATE, ABC, MFS, amino acid, sugar and inorganic solute transporters, showed altered expression profiles in response to treatments. Overall, significant differences were found in the signaling compound-elicited expression profiles of genes in roots vs those in leaves. These differences could be correlated to the underground nature of roots and their exposure to higher microbial inoculum rates under natural conditions. PMID- 18422896 TI - The model legume Medicago truncatula A17 is poorly matched for N2 fixation with the sequenced microsymbiont Sinorhizobium meliloti 1021. AB - Medicago truncatula (barrel medic) A17 is currently being sequenced as a model legume, complementing the sequenced root nodule bacterial strain Sinorhizobium meliloti 1021 (Sm1021). In this study, the effectiveness of the Sm1021-M. truncatula symbiosis at fixing N(2) was evaluated. N(2) fixation effectiveness was examined with eight Medicago species and three accessions of M. truncatula with Sm1021 and two other Sinorhizobium strains. Plant shoot dry weights, plant nitrogen content and nodule distribution, morphology and number were analysed. Compared with nitrogen-fed controls, Sm1021 was ineffective or partially effective on all hosts tested (excluding M. sativa), as measured by reduced dry weights and shoot N content. Against an effective strain, Sm1021 on M. truncatula accessions produced more nodules, which were small, pale, more widely distributed on the root system and with fewer infected cells. The Sm1021-M. truncatula symbiosis is poorly matched for N(2) fixation and the strain could possess broader N(2) fixation deficiencies. A possible origin for this reduction in effectiveness is discussed. An alternative sequenced strain, effective at N(2) fixation on M. truncatula A17, is Sinorhizobium medicae WSM419. PMID- 18422898 TI - A latitudinal cline and response to vernalization in leaf angle and morphology in Arabidopsis thaliana (Brassicaceae). AB - Adaptation to latitudinal patterns of environmental variation is predicted to result in clinal variation in leaf traits. Therefore, this study tested for geographic differentiation and plastic responses to vernalization in leaf angle and leaf morphology in Arabidopsis thaliana. Twenty-one European ecotypes were grown in a common growth chamber environment. Replicates of each ecotype were exposed to one of four treatments: 0, 10, 20 or 30 d of vernalization. Ecotypes from lower latitudes had more erect leaves, as predicted from functional arguments about selection to maximize photosynthesis. Lower-latitude ecotypes also had more elongated petioles as predicted by a biomechanical constraint hypothesis. In addition, extended vernalization resulted in shorter and more erect leaves. As predicted by functional and adaptive hypotheses, our results show genetically based clinal variation as well as environmentally induced variation in leaf traits. PMID- 18422899 TI - Geographical variation in the response to nitrogen deposition in Arabidopsis lyrata petraea. AB - The adaptive responses to atmospheric nitrogen deposition for different European accessions of Arabidopsis lyrata petraea were analysed using populations along a strong atmospheric N-deposition gradient. Plants were exposed to three N deposition rates, reflecting the rates at the different locations, in a full factorial design. Differences between accessions in the response to N were found for important phenological and physiological response variables. For example, plants from low-deposition areas had higher nitrogen-use efficiencies (NUE) and C : N ratios than plants from areas high in N deposition when grown at low N deposition rates. The NUE decreased in all accessions at higher experimental deposition rates. However, plants from high-deposition areas showed a limited capacity to increase their NUE at lower experimental deposition rates. Plants from low-deposition areas had faster growth rates, higher leaf turnover rates and shorter times to flowering, and showed a greater increase in growth rate in response to N deposition than those from high-deposition areas. Indications for adaptation to N deposition were found, and results suggest that adaptation of plants from areas high in N deposition to increased N deposition has resulted in the loss of plasticity. PMID- 18422900 TI - Soybean defense responses to the soybean aphid. AB - Transcript profiles in aphid (Aphis glycines)-resistant (cv. Dowling) and susceptible (cv. Williams 82) soybean (Glycine max) cultivars using soybean cDNA microarrays were investigated. Large-scale soybean cDNA microarrays representing approx. 18 000 genes or c. 30% of the soybean genome were compared at 6 and 12 h post-application of aphids. In a separate experiment utilizing clip cages, expression of three defense-related genes were examined at 6, 12, 24, 48, and 72 h in both cultivars by quantitative real-time PCR. One hundred and forty genes showed specific responses for resistance; these included genes related to cell wall, defense, DNA/RNA, secondary metabolism, signaling and other processes. When an extended time period of sampling was investigated, earlier and greater induction of three defense-related genes was observed in the resistant cultivar; however, the induction declined after 24 or 48 h in the resistant cultivar but continued to increase in the susceptible cultivar after 24 h. Aphid-challenged resistant plants showed rapid differential gene expression patterns similar to the incompatible response induced by avirulent Pseudomonas syringae. Five genes were identified as differentially expressed between the two genotypes in the absence of aphids. PMID- 18422901 TI - Variation in horizontal and vertical transmission of the endophyte Epichloe elymi infecting the grass Elymus hystrix. AB - Systemic fungal endophytes (Clavicipitaceae) of grasses reproduce sexually when the fungus forms stromata and contagious ascospores, or asexually by vertical transmission of hyphae into seeds and seedlings. Vertical transmission is predicted to favor reduced virulence compared with horizontal transmission in systems with both types of transmission. Here, variation in vertical and horizontal transmission and its potential heritability in a host grass-endophyte interaction, Elymus hystrix infected with Epichloe elymi, were examined in natural populations and two common garden experiments using field-collected host tillers and seed progeny of maternal plants with known infection phenotypes. Transmission mode exhibited year-to-year variation in field and common garden environments. In the common garden there were consistent differences among maternal plant families in stroma production and significant correlations between stroma production in the common garden and in natural populations. Transmission mode differed among maternal families, spanning a continuum from pure vertical transmission to a high proportion of stroma production and horizontal transmission potential. Vertical transmission to seeds occurred at high rates in all maternal families regardless of their stroma production. Observed patterns of variation indicate that endophyte transmission mode and correlated changes in virulence can respond to selection by biotic and abiotic factors. PMID- 18422902 TI - Gene expression in Eucalyptus branch wood with marked variation in cellulose microfibril orientation and lacking G-layers. AB - In response to gravitational stresses, angiosperm trees form tension wood in the upper sides of branches and leaning stems in which cellulose content is higher, microfibrils are typically aligned closely with the fibre axis and the fibres often have a thick inner gelatinous cell wall layer (G-layer). Gene expression was studied in Eucalyptus nitens branches oriented at 45 degrees using microarrays containing 4900 xylem cDNAs, and wood fibre characteristics revealed by X-ray diffraction, chemical and histochemical methods. Xylem fibres in tension wood (upper branch) had a low microfibril angle, contained few fibres with G layers and had higher cellulose and decreased Klason lignin compared with lower branch wood. Expression of two closely related fasciclin-like arabinogalactan proteins and a beta-tubulin was inversely correlated with microfibril angle in upper and lower xylem from branches. Structural and chemical modifications throughout the secondary cell walls of fibres sufficient to resist tension forces in branches can occur in the absence of G-layer enriched fibres and some important genes involved in responses to gravitational stress in eucalypt xylem are identified. PMID- 18422903 TI - High variation in foliage and leaf litter chemistry among 45 tree species of a neotropical rainforest community. AB - Distinct ecosystem level carbon : nitrogen : phosphorus (C : N : P) stoichiometries in forest foliage have been suggested to reflect ecosystem-scale selection for physiological strategies in plant nutrient use. Here, this hypothesis was explored in a nutrient-poor lowland rainforest in French Guiana. Variation in C, N and P concentrations was evaluated in leaf litter and foliage from neighbour trees of 45 different species, and the litter concentrations of major C fractions were also measured. Litter C ranged from 45.3 to 52.4%, litter N varied threefold (0.68-2.01%), and litter P varied seven-fold (0.009-0.062%) among species. Compared with foliage, mean litter N and P concentrations decreased by 30% and 65%, respectively. Accordingly, the range in mass-based N : P shifted from 14 to 55 in foliage to 26 to 105 in litter. Resorption proficiencies indicated maximum P withdrawal in most species, but with a substantial increase in variation in litter P compared with foliage. These data suggest that constrained ecosystem-level C : N : P ratios do not preclude the evolution of highly diversified strategies of nutrient use and conservation among tropical rainforest tree species. The resulting large variation in litter quality will influence stoichiometric constraints within the decomposer food web, with potentially far-ranging consequences on nutrient dynamics and plant-soil feedbacks. PMID- 18422904 TI - Molecular networks regulating Arabidopsis seed maturation, after-ripening, dormancy and germination. AB - The transition between dormancy and germination represents a critical stage in the life cycle of higher plants and is an important ecological and commercial trait. In this review we present current knowledge of the molecular control of this trait in Arabidopsis thaliana, focussing on important components functioning during the developmental phases of seed maturation, after-ripening and imbibition. Establishment of dormancy during seed maturation is regulated by networks of transcription factors with overlapping and discrete functions. Following desiccation, after-ripening determines germination potential and, surprisingly, recent observations suggest that transcriptional and post transcriptional processes occur in the dry seed. The single-cell endosperm layer that surrounds the embryo plays a crucial role in the maintenance of dormancy, and transcriptomics approaches are beginning to uncover endosperm-specific genes and processes. Molecular genetic approaches have provided many new components of hormone signalling pathways, but also indicate the importance of hormone independent pathways and of natural variation in key regulatory loci. The influence of environmental signals (particularly light) following after-ripening, and the effect of moist chilling (stratification) are increasingly being understood at the molecular level. Combined postgenomics, physiology and molecular genetics approaches are beginning to provide an unparalleled understanding of the molecular processes underlying dormancy and germination. PMID- 18422905 TI - Mechanisms of plant survival and mortality during drought: why do some plants survive while others succumb to drought? AB - Severe droughts have been associated with regional-scale forest mortality worldwide. Climate change is expected to exacerbate regional mortality events; however, prediction remains difficult because the physiological mechanisms underlying drought survival and mortality are poorly understood. We developed a hydraulically based theory considering carbon balance and insect resistance that allowed development and examination of hypotheses regarding survival and mortality. Multiple mechanisms may cause mortality during drought. A common mechanism for plants with isohydric regulation of water status results from avoidance of drought-induced hydraulic failure via stomatal closure, resulting in carbon starvation and a cascade of downstream effects such as reduced resistance to biotic agents. Mortality by hydraulic failure per se may occur for isohydric seedlings or trees near their maximum height. Although anisohydric plants are relatively drought-tolerant, they are predisposed to hydraulic failure because they operate with narrower hydraulic safety margins during drought. Elevated temperatures should exacerbate carbon starvation and hydraulic failure. Biotic agents may amplify and be amplified by drought-induced plant stress. Wet multidecadal climate oscillations may increase plant susceptibility to drought induced mortality by stimulating shifts in hydraulic architecture, effectively predisposing plants to water stress. Climate warming and increased frequency of extreme events will probably cause increased regional mortality episodes. Isohydric and anisohydric water potential regulation may partition species between survival and mortality, and, as such, incorporating this hydraulic framework may be effective for modeling plant survival and mortality under future climate conditions. PMID- 18422906 TI - Bioenergy from plants and the sustainable yield challenge. AB - Bioenergy from plants, particularly from perennial grasses and trees, could make a substantial contribution to alleviation of global problems in climate change and energy security if high yields can be sustained. Here, yield traits in a range of key bioenergy crops are reviewed, from which several targets for future improvement can be identified. Some are already the focus of genetically modified (GM) and non-GM approaches. However, the efficient growth strategies of perennial bioenergy crops rely on newly assimilated and recycled carbon and remobilized nitrogen in a continually shifting balance between sources and sinks. This balance is affected by biotic (e.g. pest, disease) and abiotic (e.g. drought) stresses. Future research should focus on three main challenges: changing (photo)thermal time sensitivity to lengthen the growing season without risking frost damage or limiting remobilization of nutritional elements following senescence; increasing aboveground biomass without depleting belowground reserves required for next year's growth and thus without increasing the requirement for nutrient applications; and increasing aboveground biomass without increasing water use. PMID- 18422907 TI - Physiology of the oesophageal transition zone in the presence of chronic bolus retention: studies using concurrent high resolution manometry and digital fluoroscopy. AB - Distinct contraction waves (CWs) exist above and below the transition zone (TZ) between the striated and smooth muscle oesophagus. We hypothesize that bolus transport is impaired in patients with abnormal spatio-temporal coordination and/or contractile pressure in the TZ. Concurrent high resolution manometry and digital fluoroscopy were performed in healthy subjects and patients with reflux oesophagitis; a condition associated with ineffective oesophageal contractility and clearance. A detailed analysis of space-time variations in bolus movement, intra-bolus and intra-luminal pressure was performed on 17 normal studies and nine studies in oesophagitis patients with impaired bolus transit using an interactive computer based system. Compared with normal controls, oesophagitis patients had greater spatial separation between the upper and lower CW tails [median 5.2 cm (range 4.4-5.6) vs 3.1 cm (2.2-3.7)], the average relative pressure within the TZ region (TZ strength) was lower [30.8 mmHg (28.3-36.5) vs 45.8 mmHg (36.1-55.7), P < 0.001], and the risk of bolus retention was higher (90%vs 12%; P < 0.01). The presence of bolus retention was associated with a wider spatial separation of the upper and lower CWs (>3 cm, the upper limit of normal; P < 0.002), independent of the presence of oesophagitis. We conclude that bolus retention in the TZ is associated with excessively wide spatial separation between the upper and lower CWs and lower TZ muscle squeeze. These findings provide a physio-mechanical basis for the occurrence of bolus retention at the level of the aortic arch, and may underlie impaired clearance with reflux oesophagitis. PMID- 18422909 TI - A 53-year-old woman with muscular atrophy showing hypersomnia and respiratory failure. PMID- 18422908 TI - Melanocytic medulloblastoma with ganglioneurocytomatous differentiation: a case report. AB - Melanotic or melanocytic medulloblastoma is a rare variant of medulloblastoma, especially when the tumor shows advanced neuronal differentiation. We report a case of this tumor, which developed in the cerebellar vermis in an 8-year-old girl. Initial biopsy specimens were identified as classical medulloblastoma with a high MIB1 index. Surgical removal of the tumor was performed after chemo radiotherapy, and black pigments were noticed on the tumor surface. Histologically, the tumor was composed of classical medulloblastoma with the presence of pigmented epithelial cells forming tubules and clusters. Immunohistochemically, the pigmented tumor cells were positive for S100 protein, HMB45, and MART1, indicating that the pigments were derived from melanosomes, and these features were compatible with melanocytic medulloblastoma. Interestingly, some of the non-pigmented or amelanotic tumor cells were also positive for HMB45 and S100 protein. Although the tumor showed an unusual cell combination, it was distinguished from atypical teratoid/rhabdoid tumor (AT/RT) by nuclear expression of INI1/BAF45 protein. The tumor also possessed ganglion-like cells within the neuropil matrix, which resembled small mature ganglion cells, and was consequently designated as ganglioneurocytoma. The melanotic medulloblastoma and part of the ganglioneurocytomatous area were fused with each other. Hence, the present case provides new information indicating that melanocytic medulloblastoma differs from AT/RT, and that it can exhibit advanced neuronal differentiation. In addition, reduction of the tumor MIB1 index was observed after chemo radiotherapy. PMID- 18422910 TI - An immunohistochemical study on a tetanus fatal case using toxin fragment C (TTC). Should it be a useful diagnostic tool? AB - A 65-year-old man fell in his garden and sustained a right pre-radial cutaneous laceration associated with a Colles' fracture. His status for tetanus immunization was uncertain; so a course of antitetanus treatment was immediately started. Two days after admission the man suddenly developed severe nucal pain, rigidity and dysphagia. A brain CT scan was negative. His condition progressively worsened and then he developed trismus. Cultures from the wound were negative for Clostridium tetani; the CSF analysis was negative. On the 9th day after admission, the man died. A presumptive clinical diagnosis of tetanus was made. Autopsy was performed 24 h after death. An immunohistochemical study was conducted with an antibody directed against tetanus toxin fragment C (TTC). By immunohistochemical evaluation, large motor neurons in the ventral horn were immunopositive for TTC. High power magnification of the ventral horn of spinal cord gray matter samples showed TTC immunoreactivity in motor neuron axons and cell bodies, using a confocal laser scanning microscope. The correct diagnosis could be established on the basis of pathological examination with TTC immunostaining. PMID- 18422911 TI - Pleuritis caused by Acremonium strictum in a patient with colon adenocarcinoma. AB - Although Acremonium strictum is environmentally widespread as opportunistic mold, it may cause infection in patients who have immunodeficiency problems. In this study, Staphylococcus aureus and A. strictum were isolated from the pleural fluid of a patient with colon adenocarcinoma. The patient did not receive antifungal therapy because the patient died after the isolation of mold. The minimal inhibitory concentrations of amphotericin B, fluconazole, ketoconazole, itraconazole, voriconazole for the A. strictum strain isolated from pleural fluid were 0.125, 256, 2 and 1.5, 0.25 mug ml(-1) respectively. In conclusion, bacteria and fungus, especially opportunistic mold, should be taken into consideration in developing pleuritis in the patients with immune-deficiency. PMID- 18422912 TI - Basidiomycete metabolites attenuate virulence properties of Candida albicans in vitro. AB - Secreted aspartic proteases (Saps) represent an important virulence factor facilitating fungal adherence. Several protease inhibitors (PIs), including HIV PIs, have been shown to reduce Candida adhesion. The aim of this study was to ascertain whether or not the recently discovered PIs Aureoquinone and Laccaridiones A and B, isolated from Basidiomycete cultures, or Bestatin, act as Sap-inhibitors and/or inhibitors of fungal adhesion. Drug effects on candidial Sap-production were determined by Sap-ELISA. Control tubes, in the absence of drug, served as positive controls, while tubes excluding both drug and proteinase induction medium were used as negative controls. Aureoquinone as well as Laccaridiones A and B, but not Bestatin, significantly inhibited Candida albicans adhesion to both epithelial and endothelial cells in a dose dependent manner and also reduced Sap-release (effects were not because of a direct interaction of the Basidiomycete metabolites with secreted Saps). Laccaridione B was consistently found to be the most effective PI tested. Interestingly, these drugs are neither fungistatic nor fungicidal at the concentrations applied. Laccaridione B may represent a promising novel type of antimycotic drug--targeting virulence factors without killing the yeast. PMID- 18422913 TI - Rhinocerebral zygomycosis caused by Saksenae vasiformis in a diabetic patient. AB - We report a rare case of invasive rhinocerebral Saksenae vasiformis infection in a 71-year-old man with type 2 diabetes mellitus ketoacidosis. Combined histological and microbiological examination of two biopsy specimens taken at four days' intervals showed broad non-septate hyphae on direct smears of biopsies and necrosis and angioinvasion by hyphae were seen in the tissue. S. vasiformis was cultured from both biopsies. The patient had a fatal outcome even though partial debridement, liposomal amphotericin B treatment and control of diabetes were instituted. Permission for necropsy was denied. Clinical, epidemiological and pathological features of the case and manifestations of S. vasiformis infections are discussed. Most cases involve cutaneous sites and only two cases of rhinocerebral infections (both with fatal outcome) have been described. The present case is the third documented case of involvement of the sinuses and the second case of S. vasiformis infection documented in Europe. PMID- 18422914 TI - In vitro susceptibility-testing in Aspergillus species. AB - Aspergillus species are the most common causes of invasive mould infections in immunocompromised patients. The introduction of new antifungal agents and recent reports of resistance emerging during treatment of Aspergillus infections have highlighted the need for in vitro susceptibility-testing. Various testing procedures have been proposed, including macrodilution and microdilution, agar diffusion, disc diffusion and Etest. At present, one of the most widely used assays is the M38-A reference method for filamentous fungi, published by the Clinical Laboratory Standard Institute and the Etest. Recently, the European Committee on Antimicrobial Susceptibility-testing (EUCAST) has charged its Antifungal Susceptibility-testing Subcommittee (AFST-EUCAST) with the preparation of new guidelines for in vitro susceptibility-testing of antifungals against Aspergillus spp. (EUCAST-AFST-ASPERGILLUS) defining breakpoints. This paper reviews the available methods for antifungal susceptibility-testing in Aspergillus spp. as well as the scant data regarding the clinical implications of in vitro testing. PMID- 18422915 TI - Usefulness and pharmacokinetic study of oral terbinafine for hyperkeratotic-type tinea pedis. AB - To study and establish an optimal administration method of oral antifungal, terbinafine (TBF), for hyperkeratotic-type tinea pedis from the pharmacokinetic point of view, 20 patients with hyperkeratotic-type tinea pedis were given TBF 125 mg once daily for 4 weeks and observed over time for improvement of dermatological symptoms and mycological efficacy. Targeting five of the patients, TBF concentration in the stratum corneum was measured using the LC-MS/MS method. TBF was detected in the stratum corneum of the sole 1 week after beginning the treatment in some cases and reached its peak 1 week after the completion of the treatment with a concentration of 247.8 ng g(-1), which was approximately more than 50 times higher than its minimal inhibitory concentration against dermatophytes. TBF was not detected at 8 weeks post-treatment, although its concentration was 50.73 ng g(-1) at 6 weeks post-treatment. All cases were subjected to analysis for final total efficacy, general safety and usefulness. Its effectiveness rate (effective + markedly effective) was 95% (19/20) with no adverse reactions, including abnormal changes in the laboratory test values, in any patients. From the above, it is noted that TBF showed excellent efficacy and safety for refractory hyperkeratotic-type tinea pedis, and also it was considered as a useful drug to treat cutaneous mycosis, including hyperkeratotic-type tinea pedis, from the pharmacokinetic point of view. PMID- 18422916 TI - Comparative efficacy and tolerability of Ketomousse (ketoconazole foam 1%) and ketoconazole cream 2% in the treatment of pityriasis versicolor: results of a prospective, multicentre, randomised study. AB - Ketomousse (K), a new thermophobic formulation (ketoconazole 1%), has proven its efficacy in the treatment of dandruff, caused by the same agent as pityriasis versicolor (PV). The objective of this study was to compare the efficacy and tolerability of K thermophobic foam vs. ketoconazole cream 2% (N) in the treatment of PV. Forty-six patients (22 in K and 24 in N group) with PV involving no more than 15% of the total trunk surface were randomly assigned for treatment either with K or N once daily for 14 days. Three weeks after the completion of treatment, improvement rate and side-effects were evaluated by clinical and mycological examination (Wood's light). Follow-up was available for 81% of subjects. Complete resolution was observed in five patients (29%) in K group and in nine (47%) in N group (P = 0.291). One patient in the N group reported urticaria while no adverse events were reported for K. Both products were cosmetically acceptable with respect to feasibility of application and formulation with a preference for K. Ketomousse (1% ketoconazole) provides an equal efficacy and tolerability compared to ketoconazole cream 2%. Therefore, Ketomousse could be considered an excellent therapeutic option in the treatment of PV. PMID- 18422917 TI - Tinea capitis among primary school children in Anambra state of Nigeria. AB - Tinea capitis continues to be a common problem of infancy and childhood, particularly among those living in unhygienic crowded conditions. The prevalence and aetiology of tinea capitis have not been well defined in Anambra state of Nigeria - hence the need to undertake this study. A total of 47,723 primary school children residing in different regions of the State was screened for tinea capitis between 2002 and 2005. Of this number, 4498 (9.4%) had tinea capitis. The highest prevalence of the disease occurred in the Southern region of the state (12.6%). Schools in urban areas recorded lower prevalence of the disease. Male : female prevalence ratio of 1.6 : 1 was recorded. Tinea capitis occurred significantly more in children below 10 years of age than in those above this age (chi(2 )= 3.178; P < 0.10). Of 502 isolates recovered in culture, Microsporum audouinii was the most prevalent (42%), followed by M. ferrugineum (17%) and Trichophyton mentagrophytes (16%). Four of the samples revealed spore head of Aspergillus spp. in direct sample microscopy. A total of 29 (6%) isolates was atypical forms. The majority (91%) of children with the lesion came from families of three or more children, living in crowded apartments. Results showed that tinea capitis remains an important public health problem in Anambra state of Nigeria. PMID- 18422918 TI - Widespread, chronic, and fluconazole-resistant Trichophyton rubrum infection in an immunocompetent patient. AB - Chronic, widespread and invasive cutaneous dermatophytoses due to Trichopyhton rubrum are common in immunocompromised patients. In immunocompetent individuals, however, chronic widespread dermatophytoses are more often associated with onychomycosis and tinea pedis. We describe a 54-year-old immunocompetent female who presented with a 2-year history of extensive erythematous and hyper-pigmented scaly plaques involving the abdominal, gluteal and crural regions without concomitant tinea pedis, tinea manus or onychomycosis. The diagnosis was made by mycological examination including culture. The pathogen identified was T. rubrum. The patient had a history of resistance to systemic fluconazole and topical ketoconazole. After an 8-week therapy period with systemic itraconazole and sertaconazole nitrate cream, a near-complete clearing of all lesions was observed. Trichophyton rubrum may thus present atypical aspects in immmunocompetent patients. PMID- 18422919 TI - Outbreak of severe disseminated aspergillosis in a flock of ostrich (Struthio camelus). AB - This study was undertaken to describe clinical, mycological and histopathological findings in black neck ostriches affected with severe aspergillosis in a flock including 80 birds, near Tehran, Iran. The signs included anorexia, depression, notable weight loss, diarrhoea, severe respiratory distress and death. Grossly, the lungs showed numerous white to yellow caseous nodules and the walls of the thoracic and abdominal air sacs were thickened with inflammatory exudates containing cellular debris, necrotic masses and green mold colonies. Multiple nodules were observed in the liver, spleen and gastrointestinal tract as well. Histopathologically, there were conidial heads and fungal hyphae in the air sacs and multifocal necrotic and granulomatous lesions with septated and dichotomously branched hyphae in various tissues, which were stained with haematoxylin and eosin and Grocott's methenamine silver nitrate. Aspergillus fumigatus was isolated in various tissues taken from affected ostriches. PMID- 18422920 TI - Preanalytical conditions for broth microdilution antifungal susceptibility of Microsporum spp. AB - Dermatophytoses caused by the genus Microsporum require a long-duration therapy compared to infections caused by other genera. Treatment of these cutaneous infections includes topical and systemic antifungal agents. Tinea capitis and tinea unguium caused by M. canis and M. gypseum are the most difficult-to-treat dermatophytoses. There are few specific studies about corresponding antifungal susceptibility in vitro. Recently, the Clinical and Laboratory Standards Institute proposed the M38A document as standard to determine the minimum inhibitory concentrations (MICs) of several antifungal agents against conidium forming filamentous fungi; however, dermatophytes were not included in this document. This study aimed to contribute to continuing investigations concerning the optimal antifungal susceptibility testing conditions of Microsporum spp. to terbinafine, ciclopiroxolamine and griseofulvin. The results pointed out potato dextrose agar as the best culture medium for inducing conidia sporulation, inoculum density amounting to 1 x 10(3) conidia ml(-1), containing only microconidia, with an incubation time of 7 days at 28 degrees C and 100% growth inhibition serving as an endpoint. The minimum fungicidal concentration values were in accordance with the MICs values, showing a fungicidal activity of these drugs towards the tested strains. According to our results, in general, terbinafine was more active than griseofulvin and ciclopiroxolamine. PMID- 18422921 TI - Posaconazole as first line treatment for disseminated zygomycosis. AB - We describe the first case report of posaconazole use as first line agent in the treatment of disseminated zygomycosis with prosthetic hip joint and pulmonary involvement due to Rhizopus microsporus. This infection occurred in a heavily immunosuppressed patient with systemic lupus erythematosus. PMID- 18422922 TI - Acremonium spp. peritonitis in an infant. AB - Fungal peritonitis is a rare but serious complication in children on peritoneal dialysis. Clinical presentation of fungal peritonitis is similar to bacterial peritonitis and Candida spp. are the most common agent. Fungal peritonitis has been usually associated with high morbidity, mortality and its treatment is difficult. In this report, we present an infant with Acremonium spp. peritonitis. A 7-month-old boy with Down syndrome, congenital heart disease, pulmonary hypertension and congestive heart failure required peritoneal dialysis for his persistent pulmonary oedema and symptomatic hyponatremia. Acremonium spp. peritonitis developed while he was on extended spectrum antibiotics and fluconazole. The patient was successfully treated with peritoneal dialysis catheter removal and liposomal amphotericin B. The case was presented to draw attention to a rare cause of peritonitis -Acremonium spp. - in a paediatric patient. PMID- 18422923 TI - Trichophyton rubrum autoinoculation from infected nails is not such a rare phenomenon. AB - Kerion-like lesions are usually caused by zoophilic dermatophytes. Here, we present a rare case report - an inflammatory tinea barbae due to an anthropophilic fungus (Trichophyton rubrum), which is the main pathogen of onychomycosis and tinea pedis. Probably the infection, in the presented case, spread from diseased fingernails. We do postulate that physicians should consider autoinoculation as a not so rare way of fungal infection transmission. PMID- 18422924 TI - Tinea capitis among primary school children in some parts of central Nigeria. AB - Tinea capitis is the most common superficial mycosis in children of school age. Although it is of public health importance, it is not a reportable or notifiable disease; therefore, actual prevalence figures are unknown in many endemic areas. The aim of this work was to determine the prevalence of tinea capitis among primary school children in two states in central Nigeria, highlighting the main aetiological agents of the infection and possible predisposing factors. A total of 28 505 primary school children aged between 3 and 16 years were recruited for the study, from 12 primary schools in two local government areas of Benue and Plateau States of Nigeria. Of them, 796 had lesions, which were clinically suggestive of tinea capitis out of which 248 (31.2%) were confirmed positive by microscopy and culture. Tinea capitis was more frequent in males, 194 (78.2%) than in females, 54 (21.8%). Children aged 10-14 years, followed by 5-9 years were predominantly infected, with 106 (42.7%) and 100 (40.3%) respectively. There was a significant correlation between age group and occurrence of tinea capitis in the study population at 95% confidence level (P = 0.004). Tinea capitis was significantly more frequent in Jos State (44.6%) than in Gboko State (23.2%) (t = .659; 95% confidence level). The prevalence of tinea capitis was influenced by social and cultural habits of the areas rather than by population density. The aetiological agent of tinea capitis in the study population was Trichophyton soudanense, 76 (30.6%), followed by Microsporum ferrugineum, 19 (7.7%) and Microsporum audouinii, 19 (7.7%). Differences in aetiology were observed for Gboko and Jos, except for T. soudanense, which predominated in both areas. The high prevalence of tinea capitis in the areas studied may be attributed to frequent interaction with soil and animals and low level of health education on personal and environmental hygiene. Aetiological agents varied from one geographical area to another. PMID- 18422925 TI - Cascading transcriptional effects of a naturally occurring frameshift mutation in Saccharomyces cerevisiae. AB - Gene-expression variation in natural populations is widespread, and its phenotypic effects can be acted upon by natural selection. Only a few naturally segregating genetic differences associated with expression variation have been identified at the molecular level. We have identified a single nucleotide insertion in a vineyard isolate of Saccharomyces cerevisiae that has cascading effects through the gene-expression network. This allele is responsible for about 45% (103/230) of the genes that show differential gene expression among the homozygous diploid progeny produced by a vineyard isolate. Using isogenic laboratory strains, we confirm that this allele causes dramatic differences in gene-expression levels of key genes involved in amino acid biosynthesis. The mutation is a frameshift mutation in a mononucleotide run of eight consecutive T's in the coding region of the gene SSY1, which encodes a key component of a plasma-membrane sensor of extracellular amino acids. The potentially high rate of replication slippage of this mononucleotide repeat, combined with its relatively mild effects on growth rate in heterozygous genotypes, is sufficient to account for the persistence of this phenotype at low frequencies in natural populations. PMID- 18422926 TI - Do mosquitoes filter the access of Plasmodium cytochrome b lineages to an avian host? AB - Many parasites show fidelity to a set of hosts in ecological time but not evolutionary time and the determinants of this pattern are poorly understood. Malarial parasites use vertebrate hosts for the asexual stage of their life cycle but use Dipteran hosts for the sexual stage. Despite the potential evolutionary importance of Dipteran hosts, little is known of their role in determining a parasite's access to vertebrate hosts. Here, we use an avian malarial system in Panama to explore whether mosquitoes act as an access filter that limits the range of vertebrate hosts used by particular parasite lineages. We amplified and sequenced Plasmodium mitochondrial DNA (mtDNA) from Turdus grayi (clay-coloured robin) and from mosquitoes at the same study site. We trapped and identified to species 123 141 female mosquitoes and completed polymerase chain reaction (PCR) screening for Plasmodium parasites in 435 pools of 20 mosquitoes per pool (8700 individuals total) spanning the 11 most common mosquito species. Our primers amplified nine Plasmodium lineages, whose sequences differed by 1.72%-10.0%. Phylogenetic analyses revealed partial clustering of lineages that co-occurred in mosquito hosts. However PAN3 and PAN6, the two primary parasite lineages of T. grayi, exhibited sequence divergence of 8.59% and did not cluster in the phylogeny. We detected these two lineages exclusively in mosquitoes from different genera - PAN3 was found only in Culex (Melanoconion) ocossa, and PAN6 was found only in Aedeomyia squamipennis. Furthermore, each of these two parasite lineages co-occurred in mosquitoes with other Plasmodium lineages that were not found in the vertebrate host T. grayi. Together, this evidence suggests that parasite-mosquito associations do not restrict the access of parasites to birds but instead may actually facilitate the switching of vertebrate hosts that occurs over evolutionary time. PMID- 18422927 TI - Ancestry and divergence of subtropical montane forest isolates: molecular biogeography of the genus Abies (Pinaceae) in southern Mexico and Guatemala. AB - The genus Abies has a complex history in southern Mexico and Guatemala. In this region, four closely related species, Abies flinckii, A. guatemalensis, A. hickelii, and A. religiosa, are distributed in fragmented and isolated montane populations. Range-wide genetic variation was investigated across species using cytoplasmic DNA markers with contrasted inheritance. Variation at two maternally inherited mitochondrial DNA markers was low. All species shared two of the nine mitotypes detected, while the remaining seven mitochondrial DNA types were restricted to a few isolated stands. Mitochondrial genetic differentiation across taxa was high (G(ST) = 0.933), it was not related to the taxonomic identity (amova; P > 0.05) of the populations, and it was not phylogeographically structured (G(ST) approximately N(ST)). In contrast, variation at three paternally inherited chloroplast DNA microsatellites was high. Chloroplast genetic differentiation was lower (G(ST) = 0.402; R(ST) = 0.547) than for mitochondrial DNA, but it was significantly related to taxonomy (amova; P < 0.001), and exhibited a significant phylogeographical structure (G(ST) < R(ST)). Different analyses of population structure indicated that A. flinckii was the most divergent taxon, while the remaining three species formed a relatively homogeneous group. However, a small number of the populations of these three taxa, all located at the limits of their respective ranges or in the Transverse Volcanic Belt, diverged from this main cluster. These trends suggest that the Mesoamerican Abies share a recent common ancestor and that their divergence and speciation is mainly driven by genetic drift and isolation during the warm interglacial periods. PMID- 18422928 TI - Postglacial recolonization at a snail's pace (Trochulus villosus): confronting competing refugia hypotheses using model selection. AB - The localization of Last Glacial Maximum (LGM) refugia is crucial information to understand a species' history and predict its reaction to future climate changes. However, many phylogeographical studies often lack sampling designs intensive enough to precisely localize these refugia. The hairy land snail Trochulus villosus has a small range centred on Switzerland, which could be intensively covered by sampling 455 individuals from 52 populations. Based on mitochondrial DNA sequences (COI and 16S), we identified two divergent lineages with distinct geographical distributions. Bayesian skyline plots suggested that both lineages expanded at the end of the LGM. To find where the origin populations were located, we applied the principles of ancestral character reconstruction and identified a candidate refugium for each mtDNA lineage: the French Jura and Central Switzerland, both ice-free during the LGM. Additional refugia, however, could not be excluded, as suggested by the microsatellite analysis of a population subset. Modelling the LGM niche of T. villosus, we showed that suitable climatic conditions were expected in the inferred refugia, but potentially also in the nunataks of the alpine ice shield. In a model selection approach, we compared several alternative recolonization scenarios by estimating the Akaike information criterion for their respective maximum-likelihood migration rates. The 'two refugia' scenario received by far the best support given the distribution of genetic diversity in T. villosus populations. Provided that fine-scale sampling designs and various analytical approaches are combined, it is possible to refine our necessary understanding of species responses to environmental changes. PMID- 18422929 TI - Contrasting patterns of variation in MHC loci in the Alpine newt. AB - Major histocompatibility complex (MHC) genes are essential in pathogen recognition and triggering an adaptive immune response. Although they are the most polymorphic genes in vertebrates, very little information on MHC variation and patterns of evolution are available for amphibians, a group known to be declining rapidly worldwide. As infectious diseases are invoked in the declines, information on MHC variation should contribute to devising appropriate conservation strategies. In this study, we examined MHC variation in 149 Alpine newts (Mesotriton alpestris) from three allopatric population groups in Poland at the northeastern margin of the distribution of this species. The genetic distinctiveness of the population groups has previously been shown by studies of skin graft rejection, allozymes and microsatellites. Two putative expressed MHC II loci with contrasting levels of variation and clear evidence of gene conversion/recombination between them were detected. The Meal-DAB locus is highly polymorphic (37 alleles), and shows evidence of historical positive selection for amino acid replacements and substantial geographical differentiation in allelic richness. On the contrary, the Meal-DBB locus exhibits low polymorphism (three alleles differing by up to two synonymous substitutions) and a uniform distribution of three alleles among geographical regions. The uniform frequencies of the presumptively neutral Meal-DBB alleles may be explained by linkage to Meal DAB. We found differences in allelic richness in Meal-DAB between regions, consistent with the hypothesis that genetic drift prevails with increasing distance from glacial refugia. Pseudogene loci appear to have evolved neutrally. The level of DAB variation correlated with variation in microsatellite loci, implying that selection and drift interplayed to produce the pattern of MHC variation observed in marginal populations of the Alpine newt. PMID- 18422930 TI - Do male moor frogs (Rana arvalis) lek with kin? AB - Many amphibian species are known to form leks during breeding season, yet it has seldom been tested which evolutionary forces are likely to act on lek formation in this taxon. We tested the kin selection hypothesis for lek formation by using eight variable microsatellite loci to compare the genetic relationship of 203 males in seven Rana arvalis leks. The results indicate that moor frog males do not lek with kin: their relatedness within leks was not higher than expected by chance. Furthermore, spatially distinct leks within same water bodies could not be distinguished from each other as separate units. These results are not expected if kin selection underlie lek formation. On the basis of these results and general knowledge of anuran breeding biology, we suggest that lek formation in explosively breeding amphibians might have evolved by female choice for breeding aggregations, combined with female choice of habitat. Future work should aim at predicting aggregations based on rules of phonotaxis over different spatial scales, and empirical work should document visitation rates not only for leks of a specific size, but also for different travel distances that visiting females may have had to cover. PMID- 18422931 TI - Temporal and spatial analyses disclose consequences of habitat fragmentation on the genetic diversity in capercaillie (Tetrao urogallus). AB - As a result of habitat fragmentation, the capercaillie (Tetrao urogallus) population in the Black Forest mountain range in southwestern Germany has declined rapidly during the last decades and now persists in patchy isolated fragments. To study the effects of fragmentation, we quantified dispersal patterns by genotyping 213 individuals in four subpopulations. We used a landscape genetics approach to analyse individual genetic variation, and despite overall low genetic structure, we found strong indications for a major boundary separating the northern part of the Black Forest area from the other subpopulations. Males and females display different gene flow patterns across the landscape. Females tend to disperse across longer distances than do males. We additionally studied the effects of the population decline on genetic diversity during the last hundred years. Although the population has dramatically declined from over 4000 to 250 males over a few decades, genetic diversity was not affected in the same way. We found two haplotypes that were present only in historic samples but microsatellite markers revealed no significant reduction in genetic diversity. Among historic samples, genetic differentiation was very low, indicating that the current genetic structure is caused by recent habitat fragmentation. We argue that inferences about reduced genetic diversity are drawn cautiously and recommend sampling over different temporal scales. PMID- 18422932 TI - Calibrating the avian molecular clock. AB - Molecular clocks are widely used to date phylogenetic events, yet evidence supporting the rate constancy of molecular clocks through time and across taxonomic lineages is weak. Here, we present 90 candidate avian clock calibrations obtained from fossils and biogeographical events. Cross-validation techniques were used to identify and discard 16 inconsistent calibration points. Molecular evolution occurred in an approximately clock-like manner through time for the remaining 74 calibrations of the mitochondrial gene, cytochrome b. A molecular rate of approximately 2.1% (+/- 0.1%, 95% confidence interval) was maintained over a 12-million-year interval and across most of 12 taxonomic orders. Minor but significant variance in rates occurred across lineages but was not explained by differences in generation time, body size or latitudinal distribution as previously suggested. PMID- 18422933 TI - Diverse historical processes shape deep phylogeographical divergence in the pollinating seed parasite Greya politella. AB - Understanding the historical framework in which species interactions have diversified across landscapes may help to partition the effects of vicariance and geographically variable selection in shaping the geographical mosaic of coevolving species. We used phylogeographical analyses of the pollinating seed parasite Greya politella (Lepidoptera: Prodoxidae) to define the historical processes that may have structured interactions of this species with its host plants across major biogeographical breaks in western North America. Using 648 bp of cytochrome oxidase I and amplified fragment length polymorphisims, we identified deep genetic breaks among some populations consistent with some definitions of cryptic species. A combination of phylogenetic and population genetic approaches indicates that different historical processes may have structured G. politella genetic diversity in four regions: northern Pacific Northwest, southern Oregon, southern Sierra Nevada, and the remainder of California. The northern Pacific Northwest had high genetic diversity likely due to glacial refugia and subsequent spatial expansion, concordant with some other taxa. Populations in southern Oregon possessed unique, closely related haplotypes with restricted gene flow, possibly indicating a long-standing set of populations in this endemic-rich region. Analyses of California populations showed evidence of restricted gene flow and spatial expansion with many closely related haplotypes that occupy a broad geographical range. Southern Sierra Nevada populations were genetically distinct and highly diverse, possibly due to a localized glacial refugium. Together, these results suggest that vicariance and population expansion, possibly in combination with geographically variable selection, have shaped the diversification of G. politella and its interactions with its host plants. PMID- 18422934 TI - Post-glacial history of the dominant alpine sedge Carex curvula in the European Alpine System inferred from nuclear and chloroplast markers. AB - The alpine sedge Carex curvula ssp. curvula is a clonal, dominant graminoid found in the European Alps, the Carpathians, the Pyrenees and in some of the Balkan Mountains. It is a late-successional species of acidophilous alpine meadows that occurs on sites that were covered by ice during the last glacial maximum (LGM). By applying the amplified fragment length polymorphism (AFLP) fingerprinting and chloroplast DNA (cpDNA) sequencing, we attempted to identify the recolonization routes followed by the species after the last ice retreat. We relied on the genetic diversity of 37 populations covering the entire distributional range of the species. As a wind-pollinated species, C. curvula is characterized by a low level of population genetic differentiation. Nuclear and chloroplast data both support the hypothesis of a long-term separation of Eastern (Balkans and Carpathians) and Western (Alps and Pyrenees) lineages. In the Alps, a continuum of genetic depauperation from the east to the west may be related to a recolonization wave originating in the eastern-most parts of the chain, where the main glacial refugium was likely located. The Pyrenean populations are nested within the western Alps group and show a low level of genetic diversity, probably due to recent long-distance colonization. In contrast to the Alps, we found no phylogeographical structure in the Carpathians. The combination of reduced ice extension during the Wurm period and the presence of large areas of siliceous substrate at suitable elevation suggest that in contrast to populations in the Alps, the species in the Carpathians underwent a local vertical migration rather than extinction and recolonization over long distance. PMID- 18422935 TI - Gene expression profiling of early primary biliary cirrhosis: possible insights into the mechanism of action of ursodeoxycholic acid. AB - OBJECTIVES: Primary biliary cirrhosis (PBC) is a poorly understood disease, both in terms of its pathogenesis and the mechanism of action of its most common treatment, ursodeoxycholic acid (UDCA). We used gene expression profiling to compare liver tissue from treatment-naive and UDCA-treated patients in order to outline some of the molecular changes associated with PBC and its treatment. PATIENTS AND EXPERIMENTAL DESIGN: Liver biopsy specimens from non-cirrhotic, treatment-naive (n=11) patients were compared with biopsies from UDCA-treated patients (n=20) and with 10 normal, healthy female controls. Gene expression was determined using a 19K cDNA microarray. In order to determine whether the observed changes in gene expression levels were specific to PBC or generic to liver damage overall, PBC samples were also compared with chronically diseased [48 hepatitis C virus (HCV), 18 hepatitis B virus (HBV)] and acutely stressed liver tissue (25 liver biopsies taken after reperfusion of liver transplant grafts). RESULTS: We found a gene signature specific to PBC (P or =84 mAU/mL for DCP, > or =25 ng/mL for AFP and > or =10% for AFP-L3. The sensitivity, specificity and positive predictive value (PPV) for DCP was 87%, 85% and 86.8%, for AFP 69%, 87% and 69.8%, and for AFP-L3 56%, 90% and 56.1%, respectively. DCP levels were below the ROC cut-off in all patients without HCC. In patients with single lesions, there was a direct correlation of DCP to tumor size. High levels of AFP correlated with diffuse type of HCC. All three markers were significantly elevated in the presence of metastatic HCC. No advantage was observed by combining two or three markers for HCC diagnosis. CONCLUSION: DCP had the highest sensitivity and PPV for HCC diagnosis, had a direct correlation with tumor size, and was not elevated in any patients without HCC. DCP should be used as the main serum test for HCC detection. PMID- 18422962 TI - Optimal systemic hemodynamic stability for successful clinical outcomes after adult living-donor liver transplantation: prospective observational study. AB - BACKGROUND AND AIM: Most living-donor liver transplantation (LDLT) recipients show characteristic systemic hemodynamics due to liver cirrhosis, and systemic hemodynamics after LDLT influenced postoperative graft function corresponding to outcomes. However, identities of optimal systemic hemodynamics for excellent outcomes and precise parameters for clinical strategy remain unclear. METHODS: Therefore, we performed prospective study in adult LDLT recipients from 2003. Hemodynamic parameters were prospectively recorded, and were analyzed in 40 recipients classified into three groups: cirrhotic (group I-C) or non-cirrhotic recipients (group I-NC) with good outcomes, and cirrhotic recipients (group II-C) without good outcomes. RESULTS: Group I-C retained characteristic hyperdynamics even after LDLT. However, absolute values of parameters revealed no significant differences between groups I-C and II-C, because group II-C also tended to show hyperdynamics. It is suggested that successful outcomes in cirrhotic recipients require maintenance of optimal hyperdynamic stability after LDLT, because cirrhotic vascular alterations still occurred. Because hemodynamic behaviors were different between groups I-C and I-NC, absolute values were also significantly different even in these successful two groups. Thus, absolute values themselves were not necessarily satisfactory for accurate evaluation of optimal hemodynamic stability. Cirrhotic hyperdynamics are symbolized in large blood volume (BV) circulated by high cardiac output (CO); therefore, we standardized CO against BV. CO/BV was significantly different between groups I-C and II-C, reflecting subtle variability of hyperdynamics in groups II-C, and was interestingly constant in the two successful groups. Therefore, CO/BV reliably evaluated optimal hemodynamic stability after LDLT, and accurately predicted outcomes. CONCLUSION: Identification of inappropriate hemodynamics after LDLT is advantageous to further improve LDLT outcomes. PMID- 18422964 TI - Autologous bone marrow cell infusion therapy for liver cirrhosis. AB - The plasticity of bone marrow cells (BMC) has been confirmed by autopsy results of female recipients of BMC from male donors. To establish new clinical therapies for patients with liver cirrhosis using autologous BMC, we developed a new in vivo murine model using green fluorescent protein (GFP) and repeated carbon tetrachloride (CCl(4)) injection. We found that BMC infused through the tail vein, efficiently repopulated cirrhotic liver tissue and, under the influence of persistent liver damage induced by carbon tetrachloride, differentiated into albumin-producing hepatocytes. Moreover, such BMC infusions into mice with cirrhosis improved liver function and reduced mortality. The latter observation correlated with the strong expression of matrix metalloproteinases (MMP), particularly MMP-9, and reduced hepatic fibrosis. The results from the 'GFP/CCl(4) model' showed that cell therapy using autologous BMC has the potential to become an effective treatment for patients with liver failure due to advanced liver cirrhosis. This review summarizes previous findings plus these recent experimental results, as well as recent clinical trials of BMC transfusion into patients with end-stage chronic liver disease. PMID- 18422963 TI - Polaprezinc attenuates liver fibrosis in a mouse model of non-alcoholic steatohepatitis. AB - BACKGROUND AND AIM: The effect of polaprezinc, a zinc-carnosine chelate compound, on the development of non-alcoholic steatohepatitis (NASH) was investigated in dietary methionine and choline deficient (MCD) mice. METHODS: Mice were fed the MCD diet with or without polaprezinc (2.2 g/kg diet) for 10 weeks. Liver histopathology, triglyceride and lipid peroxide levels, and the expression of genes linked to fibrosis were then assessed. RESULTS: MCD mice developed steatohepatitis accompanied by mild fibrosis with an increase in lipid peroxidation, hepatic stellate cell (HSC) activation, and the augmented mRNA expression of tumor necrosis factor-alpha, transforming growth factor-beta1 and procollagen alpha1(I). The mRNA expression levels of matrix metalloproteinase (MMP)-2 and tissue inhibitors of metalloproteinase (TIMP)-1 and TIMP-2 were also enhanced. Histopathologically, polaprezinc supplementation did not influence the development of steatosis but it apparently attenuated fibrosis. Polaprezinc slightly reduced lipid peroxidation and suppressed HSC activation as well as the mRNA expression of pro-inflammatory cytokines. Polaprezinc affected the MCD diet enhanced expression of TIMP-1 even when administered relatively late. CONCLUSION: These results suggest that polaprezinc attenuates fibrosis in NASH by reducing inflammation and lipid peroxidation and, during a later phase, promoting fibrolysis via the inhibition of TIMP expression in the liver. Further investigation is required to clarify the clinical efficacy of polaprezinc in patients with NASH. PMID- 18422965 TI - Large eddy simulation and zonal modeling of human-induced contaminant transport. AB - An immersed boundary method for particulate flow in an Eulerian framework is utilized to examine the effects of complex human motion on the transport of trace contaminants. The moving human object is rendered as a level set in the computational domain, and realistic human walking motion is implemented using a human kinematics model. A large eddy simulation (LES) technique is used to simulate the fluid and particle dynamics induced by human activity. Parametric studies are conducted within a Room-Room and a Room-Hall configuration, each separated by an open doorway. The effects of the average walking speed, initial proximity from the doorway, and the initial mass loading on room-to-room contaminant transport are examined. The rate of mass transport increases as the walking speed increases, but the total amount of material transported is more influenced by the initial proximity of the human from the doorway. The Room-Hall simulations show that the human wake transports material over a distance of about 8 m. Time-dependent data extracted from the simulations is used to develop a room averaged zonal model for contaminant transport due to human walking motion. The model shows good agreement with the LES results. PRACTICAL IMPLICATIONS: The effect of human activity on contaminant transport may be important in applications such as clean or isolation room design for biochemical production lines, in airborne infection control, and in entry/exit into collective protection or decontamination systems. The large eddy simulations (LES) performed in this work allow precise capturing of the local wakes generated by time dependent human motion and thus provide a means of quantifying contaminant transport due to wake effects. The LES database can be used to develop zonal models for the bulk effects of human-induced contaminant transport. These may be incorporated into multi-zone infiltration models for use in threat-response and exposure mitigation studies. PMID- 18422966 TI - Structural flexibility of the methanogenic-type seryl-tRNA synthetase active site and its implication for specific substrate recognition. AB - Seryl-tRNA synthetase (SerRS) is a class II aminoacyl-tRNA synthetase that catalyzes serine activation and its transfer to cognate tRNA(Ser). Previous biochemical and structural studies have revealed that bacterial- and methanogenic type SerRSs employ different strategies of substrate recognition. In addition to other idiosyncratic features, such as the active site zinc ion and the unique fold of the N-terminal tRNA-binding domain, methanogenic-type SerRS is, in comparison with bacterial homologues, characterized by a notable shortening of the motif 2 loop. Mutational analysis of Methanosarcina barkeri SerRS (mMbSerRS) was undertaken to identify the active site residues that ensure the specificity of amino acid and tRNA 3'-end recognition. Residues predicted to contribute to the amino acid specificity were selected for mutation according to the crystal structure of mMbSerRS complexed with its cognate aminoacyl-adenylate, whereas those involved in binding of the tRNA 3'-end were identified and mutagenized on the basis of modeling the mMbSerRS:tRNA complex. Although mMbSerRSs variants with an altered serine-binding pocket (W396A, N435A, S437A) were more sensitive to inhibition by threonine and cysteine, none of the mutants was able to activate noncognate amino acids to greater extent than the wild-type enzyme. In vitro kinetics results also suggest that conformational changes in the motif 2 loop are required for efficient serylation. PMID- 18422967 TI - Molecular basis of perinatal hypophosphatasia with tissue-nonspecific alkaline phosphatase bearing a conservative replacement of valine by alanine at position 406. Structural importance of the crown domain. AB - Hypophosphatasia, a congenital metabolic disease related to the tissue nonspecific alkaline phosphatase gene (TNSALP), is characterized by reduced serum alkaline phosphatase levels and defective mineralization of hard tissues. A replacement of valine with alanine at position 406, located in the crown domain of TNSALP, was reported in a perinatal form of hypophosphatasia. To understand the molecular defect of the TNSALP (V406A) molecule, we examined this missense mutant protein in transiently transfected COS-1 cells and in stable CHO-K1 Tet-On cells. Compared with the wild-type enzyme, the mutant protein showed a markedly reduced alkaline phosphatase activity. This was not the result of defective transport and resultant degradation of TNSALP (V406A) in the endoplasmic reticulum, as the majority of newly synthesized TNSALP (V406A) was conveyed to the Golgi apparatus and incorporated into a cold detergent insoluble fraction (raft) at a rate similar to that of the wild-type TNSALP. TNSALP (V406A) consisted of a dimer, as judged by sucrose gradient centrifugation, suggestive of its proper folding and correct assembly, although this mutant showed increased susceptibility to digestion by trypsin or proteinase K. When purified as a glycosylphosphatidylinositol-anchorless soluble form, the mutant protein exhibited a remarkably lower Kcat/Km value compared with that of the wild-type TNSALP. Interestingly, leucine and isoleucine, but not phenylalanine, were able to substitute for valine, pointing to the indispensable role of residues with a longer aliphatic side chain at position 406 of TNSALP. Taken together, this particular mutation highlights the structural importance of the crown domain with respect to the catalytic function of TNSALP. PMID- 18422968 TI - Lipid-induced conformational transition of the amyloid core fragment Abeta(28-35) and its A30G and A30I mutants. AB - The interaction of the beta-amyloid peptide (Abeta) with neuronal membranes could play a key role in the pathogenesis of Alzheimer's disease. Recent studies have focused on the interactions of Abeta oligomers to explain the neuronal toxicity accompanying Alzheimer's disease. In our study, we have investigated the role of lipid interactions with soluble Abeta(28-35) (wild-type) and its mutants A30G and A30I in their aggregation and conformational preferences. CD and Trp fluorescence spectroscopic studies indicated that, immediately on dissolution, these peptides adopted a random coil structure. Upon addition of negatively charged 1,2 dipalmitoyl-syn-glycero-3-phospho-rac-(glycerol) sodium salt (PG) lipid, the wild type and A30I mutant underwent reorganization into a predominant beta-sheet structure. However, no conformational changes were observed in the A30G mutant on interaction with PG. In contrast, the presence of zwitterionic 1,2-dipalmitoyl syn-glycero-3-phosphatidylcholine (PC) lipid had no effect on the conformation of these three peptides. These observations were also confirmed with atomic force microscopy and the thioflavin-T assay. In the presence of PG vesicles, both the wild-type and A30I mutant formed fibrillar structures within 2 days of incubation in NaCl/P(i), but not in their absence. Again, no oligomerization was observed with PC vesicles. The Trp studies also revealed that both ends of the three peptides are not buried deep in the vesicle membrane. Furthermore, fluorescence spectroscopy using the environment-sensitive probe 1,6-diphenyl-1,3,5-hexatriene showed an increase in the membrane fluidity upon exposure of the vesicles to the peptides. The latter effect may result from the lipid head group interactions with the peptides. Fluorescence resonance energy transfer experiments revealed that these peptides undergo a random coil-to-sheet conversion in solution on aging and that this process is accelerated by negatively charged lipid vesicles. These results indicate that aggregation depends on hydrophobicity and propensity to form beta-sheets of the amyloid peptide, and thus offer new insights into the mechanism of amyloid neurodegenerative disease. PMID- 18422969 TI - Toll-like receptor 4 mediates cross-talk between peroxisome proliferator activated receptor gamma and nuclear factor-kappaB in macrophages. AB - The peroxisome proliferator-activated receptor gamma (PPARgamma) is expressed in macrophages and plays an important role in suppressing the inflammatory response. Lipopolysaccharides (LPS), which activate Toll-like receptor 4 (TLR4), reduced PPARgamma expression and function in peritoneal macrophages and macrophage cell lines. Moreover, pretreatment with the synthetic PPARgamma ligand, rosiglitazone did not prevent LPS-mediated downregulation of PPARgamma. Inhibition of PPARgamma expression was not blocked by cycloheximide, indicating that de novo protein synthesis is not required for LPS-mediated suppression of PPARgamma. Destabilization of PPARgamma messenger RNA (mRNA) was not observed in LPS stimulated macrophages, suggesting that LPS regulates the synthesis of PPARgamma mRNA. LPS had no effect on PPARgamma expression in macrophages from TLR4 knockout mice, whereas LPS inhibited PPARgamma expression in cells that had been reconstituted to express functional TLR4. Targeting the TLR4 pathway with inhibitors of MEK1/2, p38, JNK and AP-1 had no effect on PPARgamma downregulation by LPS. However, inhibitors that target NEMO, IkappaB and NF-kappaB abolished LPS mediated downregulation of PPARgamma in LPS-stimulated macrophages. Our data indicate that activation of TLR4 inhibits PPARgamma mRNA synthesis by an NF kappaB-dependent mechanism. Low-density genomic profiling of macrophage-specific PPARgamma knockout cells indicated that PPARgamma suppresses inflammation under basal conditions, and that loss of PPARgamma expression is sufficient to induce a proinflammatory state. Our data reveal a regulatory feedback loop in which PPARgamma represses NF-kappaB-mediated inflammatory signalling in unstimulated macrophages; however, upon activation of TLR4, NF-kappaB drives down PPARgamma expression and thereby obviates any potential anti-inflammatory effects of PPARgamma in LPS-stimulated macrophages. PMID- 18422970 TI - State and trait anxiety and depression in patients affected by gastrointestinal diseases: psychometric evaluation of 1641 patients referred to an internal medicine outpatient setting. AB - OBJECTIVES: To evaluate state and trait form of anxiety and current depression in patients affected by gastrointestinal diseases. METHODS: We studied 1641 outpatients with gastrointestinal disorders, consecutively referred to our Internal Medicine outpatients from 1997 to 2005. State and trait anxiety were assessed by the State and Trait Anxiety Inventory. Current depression was assessed by the Zung self-rating depression scale. RESULTS: Among patients, 1379 (84.1%) showed state anxiety, 1098 (67%) showed trait anxiety and 442 (27%) showed current depression. The number of gastrointestinal diseases was directly correlated to state anxiety (p < 0.001) and trait anxiety (p = 0.04). Females showed higher levels of anxiety and depression than males (p < 0.001). State anxiety was related to food allergies (p < 0.001), small intestinal bacterial overgrowth (SIBO) (p = 0.001), Hp infection (p = 0.01) and ulcerative colitis in active phase (p = 0.03). Trait anxiety was related to irritable bowel syndrome (IBS) (p < 0.001), Helicobacter pylori (Hp) infection (p = 0.001), food allergies (p = 0.001) and SIBO (p = 0.001). Current depression was related to IBS (p < 0.001) and coeliac disease (p = 0.01), SIBO (p = 0.02). A predicted probability of 0.77 +/- 0.16 to have state anxiety, of 0.66 +/- 0.12 to have trait anxiety and of 0.39 +/- 0.14 to have depression was found in these patients. CONCLUSIONS: Most of the patients who seek medical consultation for gastrointestinal problems show an associated affective disorder. These patients should be managed by a team including gastroenterologists, psychologists and/or psychiatrists, or by a gastroenterologist having expertise in the treatment of psychological disorders. PMID- 18422971 TI - Assessment of a HER2 scoring system for gastric cancer: results from a validation study. AB - AIMS: Human epidermal growth factor receptor 2 (HER2) overexpression/amplification is implicated in the development of various solid tumour types. Validated methods and scoring systems for evaluating HER2 status exist in breast cancer, but not in gastric cancer. The aim was to establish a HER2 scoring system for gastric cancer to identify suitable patients for enrollment in a trial of trastuzumab (Herceptin) in advanced metastatic gastric cancer. METHODS AND RESULTS: Formalin-fixed paraffin-embedded gastric cancer samples were tested for HER2 status using the fluorescence in situ hybridization (FISH) pharmDxt kit (Dako Denmark A/S). Immunohistochemistry (IHC) was performed using the HercepTest (Dako). Concordance between FISH and IHC was 93.5% in 168 evaluable samples. Eleven samples were scored as FISH+ but IHC- or equivocal. CONCLUSIONS: IHC/FISH discrepancies were attributed to basolateral membranous immunoreactivity of glandular cells resulting in incomplete membranous reactivity and/or a higher rate of tumour heterogeneity in gastric cancer compared with breast cancer. With modifications to the IHC scoring system, the HercepTest is considered valid for the identification of HER2+ gastric tumours for this clinical trial. Correlation of HER2 scores with clinical outcomes will be needed to determine which patients might benefit from trastuzumab therapy. PMID- 18422972 TI - Morphological classification and definition of benign, preneoplastic and non invasive neoplastic lesions of the urinary bladder. AB - The morphological classification used in this essay has been based on the most recent World Health Organization (WHO) classification of tumours of the urinary system (i.e. 2004 WHO classification). It includes epithelial abnormalities and metaplasias as well as dysplasias and carcinomas in situ. The lesions are broadly subdivided into two major groups: benign, preneoplastic and non-invasive neoplastic lesions of the urothelium; and benign, preneoplastic and non-invasive neoplastic bladder lesions other than urothelial. Each of these lesions is defined with strict morphological criteria to provide more accurate information to urologists and oncologists in managing patients. There is still debate in the literature as to whether the 2004 WHO system should be the only one to be used and whether the 1973 WHO system should be abandoned. PMID- 18422973 TI - Primary cutaneous Langerhans cell sarcoma without Birbeck granules: indeterminate cell sarcoma? AB - An 88-year-old white male presented with a rapidly growing skin nodule on the scalp. Clinically, the nodule did not appear unusual for an ordinary cutaneous neoplasm on sun-exposed skin of an elderly white male. Histopathological examination showed sheet-like epithelioid tumor cell growth with a vaguely nested pattern and frank malignant features, resembling malignant melanoma. However, the tumor cells possessed irregularly convoluted nuclei with nuclear groves, frequent multinucleation and fine vesicular cytoplasm, features highly suggestive of histiocytes. Immunohistochemistry studies showed that the tumor cells were diffusely positive for S-100 protein and CD1a and negative for HMB-45, Melan-A, cytokeratin and CD30. The provisional diagnosis of Langerhans cell sarcoma was thus favored. To confirm this diagnosis, electron microscopic examination was performed. Although classic features of histiocytes were readily identifiable, no Birbeck granules could be found upon a thorough search on repeated sections. These results are indicative of the indeterminate cell nature of the tumor. We propose a diagnosis of primary cutaneous indeterminate cell sarcoma for this unusual histiocytic neoplasm. Current classification of histiocytic neoplasms and differential diagnosis are reviewed. PMID- 18422974 TI - Distribution of muscarinic receptor subtype M3 in melanomas and their metastases. AB - BACKGROUND: Muscarinic acetylcholine receptors (mR) are involved in the regulation of cancer cell motility and cancer progression. mR have been shown in melanoma cell lines and cryostat sections of melanomas. To substantiate the experimental data, here the correlation of mR-expression with invasive growth was studied on the cellular level by comparison with HMB-45 immunoreactivity. METHODS: mR were detected by a M3 subtype-specific polyclonal antibody in normal skin, benign compound nevi, primary melanomas [nodular type, nodular malignant melanoma (NMM)] and metastases, and were compared with HMB-45 staining in parallel paraffin sections. RESULTS: The general staining pattern of anti-M3 and HMB-45 was similar with accentuation of zones with infiltrative growth. On the cellular level, only a subpopulation of the HMB-45 positive melanoma cells expressed mR. Immunoreactivity was encountered in 3 of 15 nevi, in 9 of 14 NMM and in 10 of 14 melanoma metastases. Polymorphonuclear granulocytes also exhibited strong reactivity for anti-M3. CONCLUSION: mR-expression is associated with invasive migration of melanomas. PMID- 18422975 TI - The expression of molecular mediators in the idiopathic cutaneous calcification and ossification. AB - BACKGROUND: Idiopathic cutaneous calcification and ossification occur in the absence of an abnormal serum calcium level or pre-existing tissue abnormality. The pathogenesis has not been fully elucidated. The aim of this study was to investigate the expression of several molecular mediators in the idiopathic cutaneous calcification and ossification. METHODS: Immunohistochemical study was used to evaluate the expression of molecular mediators, bone morphogenetic protein 4 (BMP-4), beta-catenin, osteopontin, osteonectin and osteocalcin, and cell markers, smooth muscle actin, CD29 and CD44. And confocal laser scanning microscopy was used to evaluate the colocalization of BMP-4 and BMP receptor type IA. RESULTS: BMP-4, beta-catenin, osteopontin, osteonectin and osteocalcin were expressed on the calcified and ossified tissue. Especially, BMP-4 was expressed on the surrounding mesenchymal cells. Smooth muscle actin positive mesenchymal cells were on around the immature ossified tissue. Mesenchymal stem cell markers, CD29 and CD44 were not expressed. CONCLUSION: Our data suggest that BMP-4, beta catenin, osteopontin, osteonectin and osteocalcin may be involved in the idiopathic cutaneous calcification and ossification. And smooth muscle actin positive mesenchymal cells may be involved in the cutaneous ossification. This study suggests that the idiopathic cutaneous calcification and ossification is highly complicated and regulated active process like ectopic calcification of other tissues. PMID- 18422976 TI - Desmocollin expression in oral atrophic lichen planus correlates with clinical behavior and DNA content. AB - BACKGROUND: Oral lichen planus (OLP) is a chronic mucocutaneous inflammatory disease, with some tendency toward malignant transformation. Markers are needed to identify the lesions at risk. METHODS: A series of 82 biopsies from 70 patients with atrophic OLP was analyzed for desmocollin-1, E-cadherin, cyclin dependent kinase 1 (cdk-1) and Rad-51 expression using immunohistochemistry and static DNA cytometry, with particular reference to clinical outcome. RESULTS: Desmocollin-1 and E-cadherin expression were each detected in 24.4% (20/82) of the samples. Of the positive samples, only eight specimens expressed both desmocollin-1 and E-cadherin. Strong desmocollin-1 and E-cadherin expression was found in 8.5% and 3.7% of OLP biopsies, respectively. Desmocollin-1 expression increased the risk of dysplasia 31.8-fold (95% confidence intervals (CI) 3.6 280.9; p = 0.0001), while E-cadherin was significantly related to cancer (odds ratio (OR) = 5.13; 95% CI 3.3-8.1; p = 0.001). In univariate survival analysis, desmocollin-1 was a significant predictor of both cancer (log-rank test; p = 0.033) and dysplasia (p = 0.0001), while E-cadherin predicted the development of cancer (p = 0.0001). Neither cdk-1 nor Rad-51 had any predictive value. Importantly, desmocollin-1 retained its value as the only independent predictor of dysplasia in the multivariate (Cox) model (adjusted Hazard Ratio (HR) = 44.13; 95% CI 3.7-525.6). CONCLUSIONS: In atrophic OLP, desmocollin-1 is a powerful predictor of an important intermediate endpoint marker (dysplasia) in the causal pathway toward oral cancer. PMID- 18422977 TI - Extramammary Paget's disease presenting as alopecia neoplastica. AB - Extramammary Paget's disease (EMPD) is a rare malignant neoplasm with a predilection for the apocrine-rich anogenital skin and less commonly for the axilla. The tumor rarely occurs in non-apocrine bearing regions where it is referred to as ectopic EMPD. Here, we report a case of a patient that presented with a poorly circumscribed, erythematous plaque with patchy alopecia on the scalp. Histology showed pagetoid infiltration of the epidermis by atypical single and nested cells with abundant amphophilic cytoplasm with nuclear hyperchromasia, extending along the adnexae. Immunohistochemistry showed that the tumor cells stained positively for mucicarmine, periodic acid schiff, cytokeratin-7, polyclonal carcinoembryonic antigen, epithelial membrane antigen, gross cystic disease fluid protein, androgen receptor and Her-2-neu; and negatively for S-100, HMB-45, CDX-2, thyroid transcription factor-1, estrogen receptor and progesterone receptor, thus, establishing the diagnosis of ectopic EMPD. Subsequent workup showed no underlying malignancy. To our knowledge, we report the second case, and the first in the English literature, of an ectopic EMPD involving the scalp without any associated malignancy. PMID- 18422978 TI - Pedunculated hemangiopericytoma-like tumor: peculiar fibroepithelial polyp or fibrous histiocytoma variant. AB - Pathologists are continually challenged with the difficult task of discriminating between innocuous disease processes and potentially malignant entities. Apropos of this concern, we present a series of three cutaneous polypoid lesions that simulated fibroepithelial polyp, yet upon close scrutiny yielded histologic features of solitary fibrous tumor (SFT) or hemangiopericytoma. These pedunculated lesions showed a storiform pattern of spindled cells with interspersed gaping vascular channels reminiscent of SFT or hemangiopericytoma. Interestingly, the immunohistochemical staining profile of these lesions was negative for CD34 and positive for bcl-2 and factor XIIIa. These findings were discordant with SFT and suggest a relationship with fibrous histiocytoma. We propose that this entity represents a hitherto described variant of fibrous histiocytoma known as pedunculated hemangiopericytoma-like fibrous histiocytoma. PMID- 18422979 TI - Alcohol dependence is associated with reduced plasma and fundic ghrelin levels. AB - BACKGROUND: Conflicting data concerning the involvement of ghrelin in the pathophysiology of alcohol dependence have been reported. The aim of this study is to investigate how chronic alcohol ingestion influences plasma ghrelin levels and whether potential changes observed in plasma relate to modifications in ghrelin production in the stomach where this peptide is primarily synthesized. MATERIALS AND METHODS: Fifty-one consecutive alcoholics admitted for alcohol withdrawal were prospectively enrolled and compared to a control group of 32 healthy volunteers matched for age, sex, height and weight. All subjects underwent fasting plasma ghrelin determination. Twenty-seven randomly selected alcoholics and 17 controls underwent gastroscopy for fundic and duodenal biopsies. Tissues were fixed for histology or frozen in liquid nitrogen for ghrelin protein and mRNA determinations by a radioimmunoassay and quantitative polymerase chain reaction, respectively. Alcohol consumption was normalized to body weight (BW) or body mass index (BMI) given the influence of BW and volume distribution on alcohol levels. RESULTS: Plasma and fundic ghrelin protein levels were significantly decreased in alcoholics. Fundic but not plasma ghrelin protein levels inversely correlated with alcohol consumption normalized to BW or BMI. Ghrelin mRNA levels in fundic biopsies were similar in alcoholics and controls. No significant differences in duodenal ghrelin protein and mRNA levels were found between both groups. CONCLUSIONS: Alcoholism was associated with decreased plasma ghrelin levels partly due to reduced ghrelin production in the stomach. Alcohol affected ghrelin production on the post-transcriptional level in the fundus, whereas duodenal ghrelin secretion did not respond in a similar manner to alcohol intake. PMID- 18422980 TI - No long-term effect two years after intrauterine exposure to dexamethasone on dentate gyrus volume, neuronal proliferation and differentiation in common marmoset monkeys. AB - Glucocorticoids are prenatally administered to promote the maturation of the lungs. They, however, can affect neuronal proliferation and differentiation. In newborn marmoset monkeys, intrauterine hyperexposure to dexamethasone (DEX) resulted in a significantly decreased proliferation rate in the hippocampal dentate gyrus without affecting neuronal differentiation. In this study, marmoset monkeys received 5 mg/kg body weight DEX either during early (days 42-48) or late (days 90-96) pregnancy. The volume of the dentate granule cell layer as well as the proliferation and neuronal differentiation in the dentate gyrus of their 2 year-old offspring were investigated. The density of proliferating cells (Ki-67), apoptotic cells (in situ tailing) and cells differentiating to neurons (double cortin, TUC-4 and calretinin) were determined immunohistochemically. Analysis of the dentate granule cell layer volume showed no significant differences between early or late DEX-exposed marmosets and untreated control animals. Similarly, proliferation and neuronal differentiation in DEX-treated animals was not significantly different in comparison with controls. In summary, the decreased proliferation rate observed in newborn marmosets after intrauterine exposure to DEX was no longer detectable in their 2-year-old siblings suggesting no long lasting effect of prenatal hyperexposure to DEX on neuronal proliferation and differentiation in the dentate gyrus of marmoset monkeys. PMID- 18422981 TI - Immediate non-occlusal vs. early loading of dental implants in partially edentulous patients: a multicentre randomized clinical trial. Peri-implant bone and soft-tissue levels. AB - OBJECTIVES: To compare peri-implant bone and soft-tissue levels of immediately non-occlusally loaded vs. non-submerged early loaded implants in partially edentulous patients up to 14 months after placement. MATERIAL AND METHODS: Fifty two patients were randomized in five Italian private practices: 25 in the immediately loaded group and 27 in the early loaded group. To be immediately loaded, single implants had to be inserted with a torque of > or = 30 N cm, and splinted implants with a torque of > or = 20 N cm. Immediately loaded implants were provided with non-occluding temporary restorations within 48 h. After 2 months, the provisional restorations were placed in full occlusion. Implants were early loaded after 2 months. Final restorations were provided 8 months after placement. Blinded assessors evaluated peri-implant bone and soft-tissue levels. RESULTS: Fifty-two implants were immediately loaded and 52 were early loaded. No drop-out occurred. One single immediately loaded implant failed 2 months after placement. Both groups gradually lost peri-implant bone in a highly statistically significant manner at 2, 8, and 14 months. After 14 months, patients of both groups lost an average of 1.1 mm of peri-implant bone. There were no statistically significant differences between the two loading strategies for peri implant bone and soft-tissue level changes (P > 0.05). After 14 months, the position of the soft tissues did not change significantly from baseline (delivery of the final restorations 8 months after placement). CONCLUSIONS: There were no statistically or clinically significant differences between immediate and early loading of dental implants with regard to peri-implant bone and soft-tissue levels as evaluated in the present study. PMID- 18422982 TI - Progressive recovery of osseoperception as a function of the combination of implant-supported prostheses. AB - The extraction of teeth involves the elimination of extremely sensitive periodontal mechanoreceptors, which play an important role in oral sensory perception. OBJECTIVES: The aim of this study was to evaluate the recovery of interocclusal sensory perception for micro-thickness in individuals with different types of implant-supported prostheses. MATERIALS AND METHODS: Wearers of complete dentures (CDs) comprised the negative control group (group A, n=17). The experimental group consisted of wearers of prostheses supported by osseointegrated implants (Group B, n=29), which was subsequently divided into 4 subgroups: B(1) (n=5)--implant supported overdentures (ISO) occluding with CD; B(2) (n=6)--implant-supported fixed prostheses (ISFP) occluding with CD; B(3) (n=8)--wearers of maxillary and mandibular ISFP, and B(4) (n=10)--ISFP occluding with natural dentition (ND). Individuals with ND represented the positive control group (Group C, n=24). Aluminum foils measuring 10 microm, 24 microm, 30 microm, 50 microm, 80 microm, and 104 microm thickness were placed within the premolar area, adding up to 120 tests for each individual. RESULTS: The mean tactile thresholds of groups A, B1, B2, B3, B4, and C were 92 microm, 27 microm, 27 microm, 14 microm, 10 microm, and 10 microm, respectively. [Correction added after publication online 18 April 2008: in the preceding sentence 92 microm, 27 microm, 14 microm, 10 microm and 10 microm, was corrected to 92 microm, 27 microm, 27 microm, 14 microm, 10 microm and 10 microm]. The Kruskal-Wallis test revealed significant difference among groups (P<0.05). The Dunn test revealed that group A was statistically different from groups C, B(3), and B(4), and that B(1) and B(2) were statistically different from group C. CONCLUSION: Progressive recovery of osseoperception as a function of the combination of implant-supported prostheses could be observed. Moreover, ISO and/or ISFP combinations may similarly maximize the recovery of osseoperception. PMID- 18422983 TI - Bone regeneration after topical BMP-2-gene delivery in circumferential peri implant bone defects. AB - OBJECTIVES: The aims of this study were to evaluate the rate of bone formation and osseointegration after topical gene delivery with a liposomal vector system carrying bone morphogenetic protein (BMP)-2 cDNA in combination with a collagen carrier and autologous bone as a carrier in freshly created peri-implant bone defects. MATERIALS AND METHODS: Eight domestic pigs received nine calvariae defects each (10 x 7 mm). A dental implant was inserted into the centre of each defect. In the test groups, the remaining space was filled with the liposomal vector/BMP-2 complex combined with a collagen carrier (n=18) or an autologous bone graft (n=18). Control groups were collagen only (n=18) and autologous bone graft only (n=18). RESULTS: There was a significant difference in mineralisation rate in the BMP-2/bone graft (29.9%+/- 4.8 and 68.3%+/- 7.2) and bone graft only (22.6%+/- 2.6 and 49.4%+/- 13.9) groups after 7 and 28 days. Mineralisation values were also significantly higher in the BMP-2/collagen group (21.2%+/- 16.2 and 53.1%+/- 12.5) compared with the collagen-only group (8.2%+/- 7 and 41%+/- 8.1) in two different regions after 28 days. Also the bone-to-implant contact was significantly increased in the BMP-2/bone graft group after 28 days and in the BMP-2/collagen group after 7 and 28 days compared with their control groups. CONCLUSIONS: The results of this study show a significantly positive effect of liposomal vector/BMP-2 on bone regeneration and osseointegration in bony circumferential peri-implant defects. PMID- 18422984 TI - Effect of platelet-rich plasma on bone regeneration in dentistry: a systematic review. AB - OBJECTIVE: To review systematically the reported effects of platelet-rich plasma (PRP) on bone regeneration. MATERIAL AND METHODS: Up to June 2006, MEDLINE and Cochrane databases were explored with different combinations of three search terms: 'PRP', 'bone regeneration', 'dentistry' and their synonyms. INCLUSION CRITERIA: human controlled clinical trials designed to treat maxillofacial bony defects with application of PRP (test) or without PRP (control), including at least five patients with a follow-up period of more than 3 months and using clinical assessment, radiography, histology and/or histomorphometry for evaluation. Literature search, selection of eligible articles and data extraction were carried out independently by two readers. RESULTS: The literature search revealed 108 references, of which 17 were selected for further analysis. Finally, nine articles fulfilling the inclusion criteria were selected for systematically review. Owing the substantial heterogeneity of the studies it was not possible to analyze the data statistically. An attempt was made to compare results from studies that used similar outcome measures by calculating and adding confidence intervals to the data presented in the original papers. Differences in treatment effects for periodontal defects in terms of clinical attachment level (CAL) were significant (ranging from 0.8 to 3.2 mm). The reported effects of PRP in sinus elevation (compared with their controls) were <10%. CONCLUSION: We found evidence for beneficial effects of PRP in the treatment of periodontal defects. Evidence for beneficial effects of PRP in sinus elevation appeared to be weak. No conclusions can be drawn about other applications of PRP in dentistry. PMID- 18422985 TI - In vitro osteogenic potential of bone debris resulting from placement of titanium screw-type implants. AB - OBJECTIVES: Histological sections of preclinical animal studies have shown that new bone formation around dental implants frequently contains debris derived from the original bone. Whether such debris by itself is osteogenic, i.e., can influence the process of new bone formation after implant placement, is still unknown. We therefore aim to investigate if such debris does have osteogenic potential. MATERIAL AND METHODS: To substantiate our hypothesis, we performed an in vitro study in which titanium screws were placed into animal bone, and immediately removed. Without additional treatment these were placed into a cell culture medium provided with beta-glycerophosphate. We used Von Kossa staining and scanning electron microscopy to examine calcifications on the surface of the implants. Additionally, total DNA analysis, alkaline phosphates activity, and calcium content were assessed on the screw surface. RESULTS: Light and electron microscopy revealed the increasing presence of calcified matter on the implant surface. DNA amounts doubled from days 1 to 6, while alkaline phosphatase activity and calcium content showed a pronounced increase over the entire incubation time. None of these phenomena occurred on the control samples in the same medium. CONCLUSIONS: From these results, we proved that bone debris, which arises from dental implant placement, and which is moved along by the inherent roughness of the implant, by itself has osteogenic potential. PMID- 18422986 TI - Behaviour of human osteoblastic cells cultured on plasma-sprayed titanium implants in the presence of nicotine. AB - OBJECTIVES: The aim of this work was to analyse the behaviour of human bone marrow osteoblastic cells cultured on the surface of routinely used plasma sprayed titanium implants in the presence of plasmatic and salivary nicotine levels reported in smokers. MATERIAL AND METHODS: Human bone marrow cells (first subculture) were seeded on titanium implants and cultured for 35 days in alpha minimal essential medium supplemented with 10% foetal bovine serum, 50 microg/ml ascorbic acid, 10 mM beta-glycerophosphate and 10 nM dexamethasone. Seeded implants were exposed to nicotine, 10-1 mg/ml, from days 1 to 35, and characterized for cell morphology, viability/proliferation, alkaline phosphatase (ALP) activity and matrix mineralization. RESULTS: Low levels of nicotine, 10 and 50 ng/ml, representative of the plasma concentrations reported in smokers, did not cause significant effects in the cell behaviour, although a small induction in cell growth and functional activity appeared to occur. Higher nicotine levels, 0.01-1 mg/ml, within those attained in saliva through tobacco use, caused evident dose-dependent effects in osteoblastic cell behaviour, i.e., a stimulatory effect in cell growth, ALP activity and matrix mineralization, at concentrations up to 0.2 mg/ml, and a deleterious effect at higher levels. CONCLUSIONS: Considering the high tissue diffusion potential of nicotine, the results suggest the possibility of a direct modulation of the osteoblast activity as a contributing factor to the overall effect of nicotine in the bone microenvironment around dental implants. PMID- 18422987 TI - Influence of homeopathic treatment with comfrey on bone density around titanium implants: a digital subtraction radiography study in rats. AB - OBJECTIVE: The objective of this study was to evaluate the influence of homeopathic treatment with comfrey (Shymphytum officinalis 6CH) on radiographic bone density and area around titanium implants. MATERIAL AND METHODS: Forty-eight rats were divided into two groups of 24 animals each: a control group (C) and a test group (SO). Each animal received one titanium micro-implant placed in the tibia. The animals in Group SO were subjected to 10 drops of comfrey 6CH per day mixed into their drinking water until the day of sacrifice. Eight animals of each group were sacrificed at 7, 14 and 28 days post-surgery, respectively. Standardized digital radiographs were obtained on the day of implant installation (baseline images) and on the day of sacrifice (final images). Digital subtraction of the two corresponding images was performed to evaluate changes in bone density and the area related to change around the implant between baseline and final images. RESULTS: Subtraction images demonstrated that a significant difference existed in mean shade of gray at 14 days post-surgery between Group SO (mean 175.3+/-14.4) and Group C (mean 146.2+/-5.2). Regarding the area in pixels corresponding to the bone gain in Group SO, the differences observed between the sacrifice periods and groups were only significant at 7 days sacrifice between Group SO (mean 171.2+/-21.9) and Group C (mean 64.5+/-60.4). CONCLUSION: Within the limits of this study, comfrey administration promotes an increase in radiographic bone density around titanium implants in the initial period of bone healing. PMID- 18422988 TI - Evaluation on the movement of endosseous titanium implants under continuous orthodontic forces: an experimental study in the dog. AB - OBJECTIVES: The purpose of this study was to evaluate the movement of pure titanium implants under different continuous forces in the edentulous alveolar ridge. MATERIAL AND METHODS: Four pairs of titanium implants were inserted into the right maxillary and mandibular post-extraction edentulous ridge of the experimental dog. Three different levels of continuous force (100, 200, and 500 g) were loaded onto three pairs of adjacent implant abutments using a memory Ni Ti coil spring for up to 6 months and the remaining two implant abutments as the control group received no force. The positions of implant abutments were observed and the distances between the implants abutment at the top, middle and base levels were measured at the 0th, 2nd, 3rd, 6th and 8th month of the follow-up period. RESULTS: There was no significant change in the distances between adjacent abutments loaded with 100 or 200 g continuous forces throughout the entire study period. However, significantly more movement of implant abutments was noted in the 500 g pair after the 3rd month of loading when compared with the 200 or the 100 g pair (both P < 0.001). This change further increased at the 6th month (P < 0.001, 0.01, respectively). Moreover, the difference in the measurements at the top, middle and base level indicated that the two adjacent implants moved in a tipping manner in the 500 g pair after 3 and 6 months of loading. CONCLUSION: The osseointegrated implants remained stable and rigid with a pulling force of 100 and 200 g after 6 months of loading. However, when the force reached 500 g, the implants moved in an inward-tipping pattern. The results suggested that endosseous titanium implants might not necessarily be rigid anchorages under all circumstances. PMID- 18422989 TI - Topical application of statin affects bone healing around implants. AB - OBJECTIVES: 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors (statins) are widely used for hyperlipidemia. Recent studies demonstrate that statins stimulate bone morphogenetic protein-2 expression and lead to bone formation. The aim of this study was to evaluate whether the topical application of statin enhances the osteogenesis around a titanium implant. MATERIALS AND METHODS: Ten week-old female rats received pure titanium rods in both tibiae with or without fluvastatin. Propylene glycol alginate (PGA) was used as a carrier. The rats were divided into five groups: implant-only group, implant with PGA group, low-dose group [implant+PGA containing 3 microg of fluvastatin (FS)], medium-dose group (15 microg of FS), and high-dose group (75 microg of FS). The animals were sacrificed at 1 and 2 weeks after implantation. Peri-implant bone formation was assessed by histomorphometric procedures, i.e., measuring the bone-implant contact (BIC) and peri-implant bone volume (BV). A mechanical push-out test was also performed to evaluate the implant fixation strength. Statistical differences among the groups were determined by ANOVA and P < 0.05 was considered significant. RESULTS: At week 1, there was no significant difference in BIC among the groups, however, BV and the push-out strength were significantly higher in the high-dose group than in the implant-only group. At week 2, BIC and BV had significantly increased in the high-dose group in comparison with the non-statin groups. The fluvastatin-treatment group showed a significant increase in push-out strength compared with the non-statin groups. CONCLUSION: Our histomorphometrical and mechanical evaluations revealed the positive effect of topically applied fluvastatin on the bone around the implant. PMID- 18422990 TI - 'Bone map' for a safe placement of miniscrews generated by computed tomography. AB - OBJECTIVES: The aim of this manuscript is to provide a bone map that can be used as a general guide to determine the areas where miniscrews can be safely anchored. MATERIALS AND METHODS: Twenty-one computed tomographies (CTs) of maxilla and mandible were taken by the imaging centre ORTOSCAN and two measures were taken: --first, the interradicular space in the mesiodistal plane of the maxillary and mandible teeth with slices taken at 3, 6, and 9 mm from the alveolar crest, in the palatine and vestibular slices, --second, the interradicular space in the vestibular-lingual direction. RESULTS: We can see that the greatest amount of mesiodistal bone is between the first and second inferior molars on the vestibular side and between the second and first inferior molars on the lingual side, whereas the least amount of bone on the mesiodistal plane is between the inferior incisors on the vestibular side. To determine the length of the miniscrew, we must take into account that the greatest amount of vestibular-lingual bone is between the first and second inferior molar, whereas the least amount is between the central and lateral inferior incisors. CONCLUSIONS: Mesiodistal values in vestibular are those that must be considered when inserting the microscrew so as not to damage the dental tissue. Mesiodistal measurements taken in palatal-lingual do not present problems when inserting microscrews. PMID- 18422992 TI - Particle sizes from sectional data. AB - We propose a new statistical method for obtaining information about particle size distributions from sectional data without specific assumptions about particle shape. The method utilizes recent advances in local stereology. We show how to estimate separately from sectional data the variance due to the local stereological estimation procedure and the variance due to the variability of particle sizes in the population. Methods for judging the difference between the distribution of estimated particle sizes and the distribution of true particle sizes are also provided. PMID- 18422991 TI - Bayesian case influence diagnostics for survival models. AB - We propose Bayesian case influence diagnostics for complex survival models. We develop case deletion influence diagnostics for both the joint and marginal posterior distributions based on the Kullback-Leibler divergence (K-L divergence). We present a simplified expression for computing the K-L divergence between the posterior with the full data and the posterior based on single case deletion, as well as investigate its relationships to the conditional predictive ordinate. All the computations for the proposed diagnostic measures can be easily done using Markov chain Monte Carlo samples from the full data posterior distribution. We consider the Cox model with a gamma process prior on the cumulative baseline hazard. We also present a theoretical relationship between our case-deletion diagnostics and diagnostics based on Cox's partial likelihood. A simulated data example and two real data examples are given to demonstrate the methodology. PMID- 18422993 TI - The expression of DeltaNTP73, TATP73 and TP53 genes in acute myeloid leukaemia is associated with recurrent cytogenetic abnormalities and in vitro susceptibility to cytarabine cytotoxicity. AB - TP73 encodes for two proteins: full-length TAp73 and DeltaNp73, which have little transcriptional activity and exert dominant-negative function towards TP53 and TAp73. We compared TATP73 and DeltaNTP73 expression in acute myeloid leukaemia (AML) samples and normal CD34(+) progenitors. Both forms were more highly expressed in leukaemic cells. Amongst AML blasts, TATP73 was more expressed in AML harbouring the recurrent genetic abnormalities (RGA): PML-RARA, RUNX1-RUNX1T1 and CBFB-MYH11, whereas higher DeltaNTP73 expression was detected in non-RGA cases. TP53 expression did not vary according to DeltaNTP73/TATP73 expression ratio. Leukaemic cells with higher DeltaNTP73/TATP73 ratios were significantly more resistant to cytarabine-induced apoptosis. PMID- 18422994 TI - Microvesicles in haemoglobinopathies offer insights into mechanisms of hypercoagulability, haemolysis and the effects of therapy. AB - Levels of circulating red blood cell (RBC)-derived vesicles are increased in sickle cell anaemia (SCA) and thalassaemia intermedia (TI) but the mechanisms, effects and controlling factors may differ. This study found that levels of vesicles and intravascular haemolysis were linked as shown by the correlation between levels of vesicles and plasma Hb. Vesicle levels were 6-fold greater in SCA and 4-fold greater in TI than in controls. The proportion of plasma Hb within vesicles was increased in SCA and TI with a significantly higher proportion in TI. We examined whether subpopulations of RBC expressing phosphatidylserine (PS) were a source of PS(+) vesicles and observed a significant association. Thrombin generation was promoted by the vesicles in which 40-50% expressed PS. In TI, markers of thrombin generation were significantly related to PS(+) RBC. Splenectomy in TI had significant effects including greater increases in vesicle levels, plasma Hb, PS(+) RBCs and thrombin generation markers than in unsplenectomised patients. In hydroxycarbamide (HC)-treated SCA patients these measures were decreased compared with untreated controls. The relationship between vesicle levels and plasma Hb suggests a mechanism linking vesiculation to haemolysis and consequently nitric oxide (NO) bioavailability and suggests a means by which HC treatment improves NO bioavailability. PMID- 18422995 TI - Characterization of six novel mutations in CYBA: the gene causing autosomal recessive chronic granulomatous disease. AB - One of the rarest forms of chronic granulomatous disease (CGD) is caused by mutations in CYBA, which encodes the p22-phox subunit of the phagocyte NADPH oxidase, leading to defective intracellular killing. This study investigated eight patients (six males and two females) from seven consanguineous, unrelated families with clinical CGD, positive family history and p22-phox deficiency. Mutation analysis of CYBA showed six different novel mutations: deletion of exons 3, 4 and 5; a missense mutation in exon 6 (c.373G>A); a splice site mutation in intron 5 (c.369+1G>A); a frameshift in exon 6 (c.385delGAGC); a frameshift in exon 3 (c.174delG); and a frameshift in exon 4 (c.223delC). PMID- 18422996 TI - Abundant anti-apoptotic BCL-2 is a molecular target in leukaemias with t(4;11) translocation. AB - Chemotherapy resistance from imbalanced apoptosis regulation may contribute to poor outcome in leukaemias with t(4;11). Anti-apoptotic BCL-2 expression and target modulation were characterized in cell lines with t(4;11) and BCL-2 expression was examined in MLL and non-MLL infant/paediatric leukaemia cases by Western blot analysis and/or real-time polymerase chain reaction. Cytotoxicity of Genasensetrade mark (Oblimersen Sodium, G3139) alone or combined with cytotoxic drugs was assessed by MTT [(3-4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] assays of the cell lines, applying pharmacostatistical response surface modelling of drug interactions. Apoptosis and cell cycle were evaluated by flow cytometry in RS4:11 cells. Primary leukaemias and cell lines with t(4;11) expressed abundant BCL2 mRNA and protein. Variable, sometimes substantial BCL2 mRNA was detected in other leukaemia subtypes. G3139 reduced BCL2 mRNA and protein in RS4:11 cells. The most sensitive cell line to single-agent G3139 was RS4:11. Low G3139 concentrations sensitized RS4:11 and MV4-11 cells to select anti-leukaemia cytotoxic drugs. In RS4:11 cells, combining G3139 with doxorubicin (ADR) increased active caspase 3 and TUNEL staining compared to ADR alone, indicating greater apoptosis, and G3139 increased S-phase progression. The abundant BCL-2 affords a molecular target in leukaemias with t(4;11). G3139 exhibits preclinical activity and synergy with select cytotoxic agents in RS4:11 and MV4-11 cells, and these effects occur through apoptosis. PMID- 18422997 TI - Routine bone marrow examination in the initial evaluation of paediatric Hodgkin lymphoma: the Canadian perspective. AB - Bone marrow examination (BME) in paediatric Hodgkin lymphoma (HL) was evaluated, as evidence from adult HL suggests it may be unnecessary. An internet-based survey was used to examine the practice of Canadian paediatric oncologists regarding BME in children and the impact of routine BME was evaluated in patients with HL treated at our institution. Sixteen of 17 paediatric oncology centres were represented. Forty-three percent of eligible doctors completed the survey. Routine BME for stages III and IV disease was consistent nationally. By contrast, 54% and 70% of respondents reported performing routine BME for stages I and II HL respectively. Respondents were more likely to report performing routine BME in low-stage HL if trained outside Canada (P = 0.04, stage I; P = 0.07, stage II) or practicing at smaller centres (P = 0.05, stage I; P = 0.03, stage II). At our institution, 62 patients were eligible for analysis. Only four patients (6.5%) had a positive BME. Anaemia was the only significant risk factor (P = 0.006). No patient with otherwise low stage was found to have marrow involvement. Comparison of staging with and without BME demonstrated no significant difference to final risk classification. BME in paediatric patients with low-stage HL has extremely low yield and may be unnecessary. PMID- 18422998 TI - Fibronectin bridges monocytes and reticulocytes via integrin alpha4beta1. AB - Leucocytes are emerging as critical determinants in the severity of the pathology associated with sickle cell disease (SCD) and recent studies have shown that they can bind to sickle red blood cells (SS RBCs). However, the mechanism of this interaction is unclear. The alpha4beta1 integrin on monocytes and SS reticulocytes was found to mediate the interaction of these cells in in-vitro adhesion assays and in the blood of SCD patients. Plasma fibronectin (Fn), a ligand for alpha4beta1, could link SS RBCs to monocytes, as peptides derived from both the Arg-Gly-Asp-Ser (RGDS) and CS-1 site in Fn disrupted the reticulocyte/monocyte interaction. It was further shown in whole blood that 70% of the interacting monocytes were also bound to platelets, suggesting the existence of multi-cellular aggregates in SCD. Platelet inclusion in these aggregates was mediated by a P-selectin/P-selectin glycoprotein ligand-1 interaction, which has been demonstrated to activate the monocyte. These results suggest a new model for understanding the mechanism of attachment of SS RBCs to monocytes and implicate the platelet as a component and contributor to potentially occlusive aggregates that circulate in the blood of SCD patients. PMID- 18422999 TI - Ultrastructural analysis of collagen fibrils and proteoglycans in keratoconus. AB - PURPOSE: To investigate ultrastructural alterations in the distribution of collagen fibrils (CFs) and proteoglycans (PGs) in the keratoconus cornea. METHODS: Four normal corneas (donor age 24-75 years) and four severe and one mild keratoconus corneas (donor age 24-47 years) were fixed in 2.5% glutaraldehyde containing 0.05% cuprolinic blue dye for electron microscopy. Analyses were carried out on approximately 39 000 CF and 66 000 PG filaments in the anterior, middle and posterior stroma, using analySIS soft imaging software. RESULTS: In severe keratoconus, stromal lamellae were seen to undulate in most regions, whereas in mild keratoconus only the middle and posterior lamellae were affected. In keratoconus corneas the mean diameter and interfibrillar spacing of CFs was reduced in all zones (p < 0.0001) and the CF and PG number density and area fractions were significantly increased (p < 0.0001) compared with in normal corneas and were higher (p < 0.0001) in the corneas with severe keratoconus than in that with mild keratoconus. The lamellae contained microfibrils (8-9 nm wide) and, in addition, PGs embedded within CFs. Degenerate keratocytes containing PGs were found in all keratoconus corneas. CONCLUSIONS: These studies suggest that as keratoconus progresses, the PG content of the stroma increases, whereas fibril diameter is reduced. The altered stromal content of PGs may influence CF diameters and their organization in keratoconus, weakening lateral cohesion and resulting in significant disorder of CF packing. PMID- 18423000 TI - Gene expression analyses in Atlantic salmon challenged with infectious salmon anemia virus reveal differences between individuals with early, intermediate and late mortality. AB - BACKGROUND: Infectious salmon anemia virus (ISAV) causes a multisystemic disease responsible for severe losses in salmon aquaculture. Better understanding of factors that explain variations in resistance between individuals and families is essential for development of strategies for disease control. To approach this, we compared global gene expression using microarrays in fish dying early and late in the time course following infection from a highly pathogenic ISAV. RESULTS: Tissues (gill, heart, liver and spleen) from infected Atlantic salmon (cohabitation, ISAV Glesvaer 2/90 isolate) were collected from three stages over the time course of the experiment; early (EM, 0-10% cumulative mortality (CM), 21 25 days post-infection (DPI)), intermediate (IM, 35-55% CM, 28-31 DPI) and late (LM, 75-85% CM, 37-48 DPI) mortality. Viral loads were equal in EM and IM but dropped markedly in LM fish. Gene expression analyses using a 1.8 K salmonid fish cDNA microarray (SFA2.0) and real-time qPCR revealed a large group of genes highly up-regulated across tissues in EM, which were mainly implicated in innate antiviral responses and cellular stress. Despite equal levels of MHC class I in EM and LM, increase of splenic and cardiac expression of immunoglobulin-like genes was found only in LM while a suite of adaptive immunity markers were activated already in IM. The hepatic responses to ISAV were characterized by difference between EM and LM in expression of chaperones and genes involved in eicosanoid metabolism. To develop classification of high and low resistance phenotypes based on a small number of genes, we processed results from qPCR analyses of liver using a linear discriminant analysis. Four genes (5 lipoxygenase activating protein, cytochrome P450 2K4-1, galectin-9 and annexin A1) were sufficient for correct assignment of individuals to EM, LM and uninfected groups, while IM was inseparable from EM. Three of four prognostic markers are involved in metabolism of inflammatory regulators. CONCLUSION: This study adds to the understanding of molecular determinants for resistance to acute ISAV infection. The most susceptible individuals were characterized by high viral replication and dramatic activation of innate immune responses, which did not provide protection. The ability to endure high levels of infection for sustained periods could be associated with lower inflammatory responses while subsequent protection and viral clearance was most likely conferred by activation of adaptive immunity. PMID- 18423001 TI - Population-based study of diagnostic assays for Borrelia infection: comparison of purified flagella antigen assay (Ideia, Dako Cytomation) and recombinant antigen assay (Liaison, DiaSorin). AB - BACKGROUND: Testing for Borrelia-specific IgM and IgG-antibodies are often performed on a variety of poorly defined symptoms, and isolated IgM results are a frequent finding, which results in diagnostic uncertainty and further testing. We wanted to test the hypothesis that Borrelia-specific assays using recombinant antigens perform differently from assays based on purified flagella antigen. METHODS: We compared the use of recombinant antigens (LIAISON DiaSorin, Saluggia, Italy) and purified flagella antigen (IDEIA Borrelia, DakoCytomation, Glostrup, Denmark) in the assay for Borrelia-specific IgM and IgG-antibodies. The assays were tested on an unselected population of serum samples submitted from general practice. A total of 357 consecutive samples for analysis of Borrelia IgM and IgG antibodies. Furthermore, we analysed 540 samples for Borrelia-specific IgM or IgG antibodies first by the IDEIA and, if they were positive, the samples were further analysed using the LIAISON assay. To verify the correctness of the patient's serological status, discrepant samples were analysed by line blots (EcoLine, Virotech). RESULTS: In the consecutive series of 357 samples, the IgM assays detected 308 negative and 3 positive samples with concordant results. Compared with the line blot, the IDEIA system produced 21 false-positive IgM results, whereas the LIAISON(R) system produced only one false-positive IgM result. The IgG assays showed 1 positive and 328 negative concordant results. The LIAISON system produced 9 true IgG-positive samples that were not detected by the IDEIA system, but the former produced 4 positive IgG results that were negative by line blot. CONCLUSION: Diagnostic assays based on flagella antigen seem to show more false-positive IgM and false-negative IgG results than assays based on recombinant antigens. The latter may reduce the number of presumably false positive IgM results and identify more IgG-positive subjects, but this system also produces more false-positive IgG results. PMID- 18423002 TI - Virulence studies of Enterobacter sakazakii isolates associated with a neonatal intensive care unit outbreak. AB - BACKGROUND: In 1994, an outbreak of Enterobacter sakazakii infections in France occurred in a neonatal intensive care unit during which 17 neonates were infected. More than half of the infected neonates had severe clinical symptoms; 7 cases of necrotising enterocolitis (one with abdominal perforation), one case of septicemia, and one case of meningitis. The other 8 neonates were shown to be colonized but remained asymptomatic. There were three deaths. Four distinguishable pulsotypes of E. sakazakii were isolated during the outbreak, and the deaths were attributable to one pulsotype. This paper compares strains, from the four pulsotypes, for attachment and invasion of mammalian intestinal cells, macrophage survival and blood-brain barrier invasion. A fourth death from septic shock also occurred during the E. sakazakii outbreak. This was due to E. cloacae which at the time of the outbreak had been misidentified as E. sakazakii. This isolate has been included in this study. RESULTS: All E. sakazakii strains attached and invaded Caco-2 human epithelial cells, and invaded rat brain capillary endothelial cells. The majority of strains persisted in macrophage cells for 48 h. Two strains from fatal NEC and meningitis cases showed the highest invasion rate of Caco-2 intestinal cells. Their invasion of brain capillary endothelial cells was equivalent or greater than that of the neonatal E. coli meningitis strain K1. These strains also had extended spectrum beta lactamase activities. E. cloacae differed from E. sakazakii due to the greater attachment and less invasion of epithelial cells, no survival in macrophages, and less invasion of capillary endothelial brain cells. CONCLUSION: While variables such as host factors and treatment strategies determine the outcome of infection, our in vitro studies evaluated the virulence of the isolates associated with this outbreak. It was not possible to directly correlate clinical symptoms and outcomes with in vitro studies. Nevertheless, we have shown the variation in invasive potential of E. sakazakii with intestinal and blood-brain barrier cells between and within pulsotypes from a neonatal intensive care unit outbreak. E. sakazakii strains were able to persist and even replicate for a period within macrophage cells. These traits appear to facilitate host immune evasion and dissemination. PMID- 18423003 TI - Predictors of depression stigma. AB - BACKGROUND: To investigate and compare the predictors of personal and perceived stigma associated with depression. METHOD: Three samples were surveyed to investigate the predictors: a national sample of 1,001 Australian adults; a local community sample of 5,572 residents of the Australian Capital Territory and Queanbeyan aged 18 to 50 years; and a psychologically distressed subset (n = 487) of the latter sample. Personal and Perceived Stigma were measured using the two subscales of the Depression Stigma Scale. Potential predictors included demographic variables (age, gender, education, country of birth, remoteness of residence), psychological distress, awareness of Australia's national depression initiative beyondblue, depression literacy and level of exposure to depression. Not all predictors were used for all samples. RESULTS: Personal stigma was consistently higher among men, those with less education and those born overseas. It was also associated with greater current psychological distress, lower prior contact with depression, not having heard of a national awareness raising initiative, and lower depression literacy. These findings differed from those for perceived stigma except for psychological distress which was associated with both higher personal and higher perceived stigma. Remoteness of residence was not associated with either type of stigma. CONCLUSION: The findings highlight the importance of treating the concepts of personal and perceived stigma separately in designing measures of stigma, in interpreting the pattern of findings in studies of the predictors of stigma, and in designing, interpreting the impact of and disseminating interventions for stigma. PMID- 18423004 TI - Stimulatory effect of Eucalyptus essential oil on innate cell-mediated immune response. AB - BACKGROUND: Besides few data concerning the antiseptic properties against a range of microbial agents and the anti-inflammatory potential both in vitro and in vivo, little is known about the influence of Eucalyptus oil (EO) extract on the monocytic/macrophagic system, one of the primary cellular effectors of the immune response against pathogen attacks. The activities of this natural extract have mainly been recognized through clinical experience, but there have been relatively little scientific studies on its biological actions. Here we investigated whether EO extract is able to affect the phagocytic ability of human monocyte derived macrophages (MDMs) in vitro and of rat peripheral blood monocytes/granulocytes in vivo in absence or in presence of immuno-suppression induced by the chemotherapeutic agent 5-fluorouracil (5-FU). METHODS: Morphological activation of human MDMs was analysed by scanning electron microscopy. Phagocytic activity was tested: i) in vitro in EO treated and untreated MDMs, by confocal microscopy after fluorescent beads administration; ii) in vivo in monocytes/granulocytes from peripheral blood of immuno-competent or 5-FU immuno-suppressed rats, after EO oral administration, by flow cytometry using fluorescein-labelled E. coli. Cytokine release by MDMs was determined using the BD Cytometric Bead Array human Th1/Th2 cytokine kit. RESULTS: EO is able to induce activation of MDMs, dramatically stimulating their phagocytic response. EO stimulated internalization is coupled to low release of pro-inflammatory cytokines and requires integrity of the microtubule network, suggesting that EO may act by means of complement receptor-mediated phagocytosis. Implementation of innate cell-mediated immune response was also observed in vivo after EO administration, mainly involving the peripheral blood monocytes/granulocytes. The 5-FU/EO combined treatment inhibited the 5-FU induced myelotoxicity and raised the phagocytic activity of the granulocytic/monocytic system, significantly decreased by the chemotherapic. CONCLUSION: Our data, demonstrating that Eucalyptus oil extract is able to implement the innate cell-mediated immune response, provide scientific support for an additional use of this plant extract, besides those concerning its antiseptic and anti-inflammatory properties and stimulate further investigations also using single components of this essential oil. This might drive development of a possible new family of immuno-regulatory agents, useful as adjuvant in immuno-suppressive pathologies, in infectious disease and after tumour chemotherapy. PMID- 18423005 TI - Mapping the evolution of 'food deserts' in a Canadian city: supermarket accessibility in London, Ontario, 1961-2005. AB - BACKGROUND: A growing body of research suggests that the suburbanization of food retailers in North America and the United Kingdom in recent decades has contributed to the emergence of urban 'food deserts', or disadvantaged areas of cities with relatively poor access to healthy and affordable food. This paper explores the evolution of food deserts in a mid-sized Canadian city (London, Ontario) by using a geographic information system (GIS) to map the precise locations of supermarkets in 1961 and 2005; multiple techniques of network analysis were used to assess changing levels of supermarket access in relation to neighbourhood location, socioeconomic characteristics, and access to public transit. RESULTS: The findings indicate that residents of inner-city neighbourhoods of low socioeconomic status have the poorest access to supermarkets. Furthermore, spatial inequalities in access to supermarkets have increased over time, particularly in the inner-city neighbourhoods of Central and East London, where distinct urban food deserts now exist. CONCLUSION: Contrary to recent findings in larger Canadian cities, we conclude that urban food deserts exist in London, Ontario. Policies aimed at improving public health must also recognize the spatial, as well as socioeconomic, inequities with respect to access to healthy and affordable food. Additional research is necessary to better understand how supermarket access influences dietary behaviours and related health outcomes. PMID- 18423006 TI - Empowering the people: development of an HIV peer education model for low literacy rural communities in India. AB - BACKGROUND: Despite ample evidence that HIV has entered the general population, most HIV awareness programs in India continue to neglect rural areas. Low HIV awareness and high stigma, fueled by low literacy, seasonal migration, gender inequity, spatial dispersion, and cultural taboos pose extra challenges to implement much-needed HIV education programs in rural areas. This paper describes a peer education model developed to educate and empower low-literacy communities in the rural district of Perambalur (Tamil Nadu, India). METHODS: From January to December 2005, six non-governmental organizations (NGO's) with good community rapport collaborated to build and pilot-test an HIV peer education model for rural communities. The program used participatory methods to train 20 NGO field staff (Outreach Workers), 102 women's self-help group (SHG) leaders, and 52 barbers to become peer educators. Cartoon-based educational materials were developed for low-literacy populations to convey simple, comprehensive messages on HIV transmission, prevention, support and care. In addition, street theatre cultural programs highlighted issues related to HIV and stigma in the community. RESULTS: The program is estimated to have reached over 30,000 villagers in the district through 2051 interactive HIV awareness programs and one-on-one communication. Outreach workers (OWs) and peer educators distributed approximately 62,000 educational materials and 69,000 condoms, and also referred approximately 2844 people for services including voluntary counselling and testing (VCT), care and support for HIV, and diagnosis and treatment of sexually transmitted infections (STI). At least 118 individuals were newly diagnosed as persons living with HIV (PLHIV); 129 PLHIV were referred to the Government Hospital for Thoracic Medicine (in Tambaram) for extra medical support. Focus group discussions indicate that the program was well received in the communities, led to improved health awareness, and also provided the peer educators with increased social status. CONCLUSION: Using established networks (such as community-based organizations already working on empowerment of women) and training women's SHG leaders and barbers as peer educators is an effective and culturally appropriate way to disseminate comprehensive information on HIV/AIDS to low-literacy communities. Similar models for reaching and empowering vulnerable populations should be expanded to other rural areas. PMID- 18423008 TI - Six years survival on imatinib with no disease progression after diagnosis of metastatic duodenal gastrointestinal stromal tumour: a case report. AB - INTRODUCTION: A duodenal Gastrointestinal Stromal Tumour (GIST) is a rare finding and until recently advanced disease had a poor prognosis. A PubMed search revealed no reports of more than five years survival of inoperable GIST on chemotherapy with WHO performance status zero. CASE PRESENTATION: A 68 year old female was diagnosed with unresectable GIST in the duodenum with metastasis to liver, pancreas and omentum in November 2001. She was commenced on imatinib mesylate (Glivec) chemotherapy. This case report was prepared from the medical records and radiology reports. She had good tolerance with stable disease. After six years her CT scan showed no disease progression and her WHO performance status was zero. CONCLUSION: This report supports the view that imatinib is a safe and effective drug in controlling disease progression in advanced metastatic GIST and plays an important role in improving the patient's quality of life. PMID- 18423007 TI - The IBR5 phosphatase promotes Arabidopsis auxin responses through a novel mechanism distinct from TIR1-mediated repressor degradation. AB - BACKGROUND: In Arabidopsis, INDOLE-3-BUTYRIC ACID RESPONSE5 (IBR5), a putative dual-specificity protein phosphatase, is a positive regulator of auxin response. Mutations in IBR5 result in decreased plant height, defective vascular development, increased leaf serration, fewer lateral roots, and resistance to the phytohormones auxin and abscisic acid. However, the pathways through which IBR5 influences auxin responses are not fully understood. RESULTS: We analyzed double mutants of ibr5 with other mutants that dampen auxin responses and found that combining ibr5 with an auxin receptor mutant, tir1, enhanced auxin resistance relative to either parent. Like other auxin-response mutants, auxin-responsive reporter accumulation was reduced in ibr5. Unlike other auxin-resistant mutants, the Aux/IAA repressor reporter protein AXR3NT-GUS was not stabilized in ibr5. Similarly, the Aux/IAA repressor IAA28 was less abundant in ibr5 than in wild type. ibr5 defects were not fully rescued by overexpression of a mutant form of IBR5 lacking the catalytic cysteine residue. CONCLUSION: Our genetic and molecular evidence suggests that IBR5 is a phosphatase that promotes auxin responses, including auxin-inducible transcription, differently than the TIR1 auxin receptor and without destabilizing Aux/IAA repressor proteins. Our data are consistent with the possibility that auxin-responsive transcription can be modulated downstream of TIR1-mediated repressor degradation. PMID- 18423009 TI - Melatonin therapy to improve nocturnal sleep in critically ill patients: encouraging results from a small randomised controlled trial. AB - INTRODUCTION: Sleep disturbances are common in critically ill patients and when sleep does occur it traverses the day-night periods. The reduction in plasma melatonin levels and loss of circadian rhythm observed in critically ill patients receiving mechanical ventilation may contribute to this irregular sleep-wake pattern. We sought to evaluate the effect of exogenous melatonin on nocturnal sleep quantity in these patients and, furthermore, to describe the kinetics of melatonin after oral administration in this patient population, thereby guiding future dosing schedules. METHODS: We conducted a randomised double-blind placebo controlled trial in 24 patients who had undergone a tracheostomy to aid weaning from mechanical ventilation. Oral melatonin 10 mg or placebo was administered at 9 p.m. for four nights. Nocturnal sleep was monitored using the bispectral index (BIS) and was expressed in terms of sleep efficiency index (SEI) and area under the curve (AUC). Secondary endpoints were SEI measured by actigraphy and nurse and patient assessments. Plasma melatonin concentrations were measured in nine patients in the melatonin group on the first night. RESULTS: Nocturnal sleep time was 2.5 hours in the placebo group (mean SEI = 0.26, 95% confidence interval [CI] 0.17 to 0.36). Melatonin use was associated with a 1-hour increase in nocturnal sleep (SEI difference = 0.12, 95% CI -0.02 to 0.27; P = 0.09) and a decrease in BIS AUC indicating 'better' sleep (AUC difference = -54.23, 95% CI -104.47 to 3.98; P = 0.04). Results from the additional sleep measurement methods were inconclusive. Melatonin appeared to be rapidly absorbed from the oral solution, producing higher plasma concentrations relative to similar doses reported in healthy individuals. Plasma concentrations declined biexponentially, but morning (8 a.m.) plasma levels remained supraphysiological. CONCLUSION: In our patients, nocturnal sleep quantity was severely compromised and melatonin use was associated with increased nocturnal sleep efficiency. Although these promising findings need to be confirmed by a larger randomised clinical trial, they do suggest a possible future role for melatonin in the routine care of critically ill patients. Our pharmacokinetic analysis suggests that the 10-mg dose used in this study is too high in these patients and may lead to carryover of effects into the next morning. Reduced doses of 1 to 2 mg could be used in future studies. TRIAL REGISTRATION: Current Controlled Trials ISRCTN47578325. PMID- 18423011 TI - The influence of in vitro fitness defects on pneumococcal ability to colonize and to cause invasive disease. AB - BACKGROUND: Streptococcus pneumoniae is a genetically diverse major human pathogen, yet a common colonizer of the nasopharynx. Here we analyzed the influence of defects affecting in vitro growth rate, on the ability of S. pneumoniae to colonize and to cause invasive disease in vivo. RESULTS: Of eleven different clinical isolates one serotype 14 carrier isolate showed a significantly longer generation time as compared to other isolates, and was severely attenuated in mice. To directly investigate the impact of growth rate on virulence, a panel of mutants in five non-essential housekeeping genes was constructed in the virulent TIGR4 background by insertion-deletion mutagenesis. Three of these mutants (ychF, hemK and yebC) were, to different degrees, growth defective, and showed a reduced invasiveness in an intranasal murine challenge model that correlated to their in vitro growth rate, but remained capable of colonizing the upper airways. The growth defect, as well as virulence defect of the hemK insertion-deletion mutant, was mediated by polarity effects on the downstream yrdC gene, encoding a probable chaperone in ribosome assembly. CONCLUSION: We conclude that large fitness defects are needed to completely prevent pneumococci from causing invasive disease after intranasal challenge. However, even severe growth defects still allow pneumococci to persistently colonize the upper airways. PMID- 18423012 TI - Simple tools for assembling and searching high-density picolitre pyrophosphate sequence data. AB - BACKGROUND: The advent of pyrophosphate sequencing makes large volumes of sequencing data available at a lower cost than previously possible. However, the short read lengths are difficult to assemble and the large dataset is difficult to handle. During the sequencing of a virus from the tsetse fly, Glossina pallidipes, we found the need for tools to search quickly a set of reads for near exact text matches. METHODS: A set of tools is provided to search a large data set of pyrophosphate sequence reads under a "live" CD version of Linux on a standard PC that can be used by anyone without prior knowledge of Linux and without having to install a Linux setup on the computer. The tools permit short lengths of de novo assembly, checking of existing assembled sequences, selection and display of reads from the data set and gathering counts of sequences in the reads. RESULTS: Demonstrations are given of the use of the tools to help with checking an assembly against the fragment data set; investigating homopolymer lengths, repeat regions and polymorphisms; and resolving inserted bases caused by incomplete chain extension. CONCLUSION: The additional information contained in a pyrophosphate sequencing data set beyond a basic assembly is difficult to access due to a lack of tools. The set of simple tools presented here would allow anyone with basic computer skills and a standard PC to access this information. PMID- 18423013 TI - Implication of Xenobiotic Metabolizing Enzyme gene (CYP2E1, CYP2C19, CYP2D6, mEH and NAT2) polymorphisms in breast carcinoma. AB - BACKGROUND: Xenobiotic Metabolizing Enzymes (XMEs) contribute to the detoxification of numerous cancer therapy-induced products. This study investigated the susceptibility and prognostic implications of the CYP2E1, CYP2C19, CYP2D6, mEH and NAT2 gene polymorphisms in breast carcinoma patients. METHODS: The authors used polymerase chain reaction and restriction enzyme digestion to characterize the variation of the CYP2E1, CYP2C19, CYP2D6, mEH and NAT2 gene in a total of 560 unrelated subjects (246 controls and 314 patients). RESULTS: The mEH (C/C) mutant and the NAT2 slow acetylator genotypes were significantly associated with breast carcinoma risk (p = 0.02; p = 0.01, respectively). For NAT2 the association was more pronounced among postmenopausal patients (p = 0.006). A significant association was found between CYP2D6 (G/G) wild type and breast carcinoma risk only in postmenopausal patients (p = 0.04). Association studies of genetic markers with the rates of breast carcinoma specific overall survival (OVS) and the disease-free survival (DFS) revealed among all breast carcinoma patients no association to DFS but significant differences in OVS only with the mEH gene polymorphisms (p = 0.02). In addition, the mEH wild genotype showed a significant association with decreased OVS in patients with axillary lymph node-negative patients (p = 0.03) and with decreased DFS in patients with axillary lymph node-positive patients (p = 0.001). However, the NAT2 intermediate acetylator genotype was associated with decreased DFS in axillary lymph node-negative patients. CONCLUSION: The present study may prove that polymorphisms of some XME genes may predict the onset of breast carcinoma as well as survival after treatment. PMID- 18423014 TI - Selective activation of microglia in spinal cord but not higher cortical regions following nerve injury in adult mouse. AB - Neuronal plasticity along the pathway for sensory transmission including the spinal cord and cortex plays an important role in chronic pain, including inflammatory and neuropathic pain. While recent studies indicate that microglia in the spinal cord are involved in neuropathic pain, a systematic study has not been performed in other regions of the central nervous system (CNS). In the present study, we used heterozygous Cx3cr1GFP/+mice to characterize the morphological phenotypes of microglia following common peroneal nerve (CPN) ligation. We found that microglia showed a uniform distribution throughout the CNS, and peripheral nerve injury selectively activated microglia in the spinal cord dorsal horn and related ventral horn. In contrast, microglia was not activated in supraspinal regions of the CNS, including the anterior cingulate cortex (ACC), prefrontal cortex (PFC), primary and secondary somatosensory cortex (S1 and S2), insular cortex (IC), amygdala, hippocampus, periaqueductal gray (PAG) and rostral ventromedial medulla (RVM). Our results provide strong evidence that nerve injury primarily activates microglia in the spinal cord of adult mice, and pain-related cortical plasticity is likely mediated by neurons. PMID- 18423015 TI - In vivo site-specific biotinylation of proteins within the secretory pathway using a single vector system. AB - BACKGROUND: Due to its extremely high strength, the interaction between biotin and (strept)avidin has been exploited for a large number of biotechnological applications. Site-specific biotinylation of proteins in vivo can be achieved by co-expressing in mammalian cells the protein of interest fused to a 15 amino acid long Biotin Acceptor Peptide (BAP) and the bacterial biotin-protein ligase BirA, which specifically recognizes and attaches a biotin to the single lysine residue of the BAP sequence. However, this system is mainly based on the contemporaneous use of two different plasmids or on induction of expression of two proteins through an IRES-driven mechanism. RESULTS: We developed a single bigenic plasmid that contains two independent transcriptional units for the co-expression of both the protein tagged with BAP and an engineered version of the BirA enzyme. Upstream of the cDNA encoding BirA, a signal secretion leader sequence was added to allow translocation of the enzyme to the secretory pathway. Three different recombinant antibodies in the scFv format, a membrane bound and secretory truncated IgE Fc fragment and a soluble version of the human IgE high affinity receptor were shown to be efficiently biotinylated and to maintain their binding properties in immunofluorescence microscopy, flow cytometry and ELISA assays. CONCLUSION: The present study shows the universal applicability to both secretory and membrane bound proteins of a single bigenic plasmid to induce the site specific in vivo biotinylation of target molecules tagged with a short acceptor peptide. These molecules could be easily obtained from supernatants or extracts of mammalian cells and used for a wide range of biological applications. PMID- 18423016 TI - The influence of venous admixture on alveolar dead space and carbon dioxide exchange in acute respiratory distress syndrome: computer modelling. AB - INTRODUCTION: Alveolar dead space reflects phenomena that render arterial partial pressure of carbon dioxide higher than that of mixed alveolar gas, disturbing carbon dioxide exchange. Right-to-left shunt fraction (Qs/Qt) leads to an alveolar dead space fraction (VdAS/VtA; where VtA is alveolar tidal volume). In acute respiratory distress syndrome, ancillary physiological disturbances may include low cardiac output, high metabolic rate, anaemia and acid-base instability. The purpose of the present study was to analyze the extent to which shunt contributes to alveolar dead space and perturbs carbon dioxide exchange in ancillary physiological disturbances. METHODS: A comprehensive model of pulmonary gas exchange was based upon known equations and iterative mathematics. RESULTS: The alveolar dead space fraction caused by shunt increased nonlinearly with Qs/Qt and, under 'basal conditions', reached 0.21 at a Qs/Qt of 0.6. At a Qs/Qt of 0.4, reduction in cardiac output from 5 l/minute to 3 l/minute increased VdAS/VtA from 0.11 to 0.16. Metabolic acidosis further augmented the effects of shunt on VdAS/VtA, particularly with hyperventilation. A Qs/Qt of 0.5 may increase arterial carbon dioxide tension by about 15% to 30% if ventilation is not increased. CONCLUSION: In acute respiratory distress syndrome, perturbation of carbon dioxide exchange caused by shunt is enhanced by ancillary disturbances such as low cardiac output, anaemia, metabolic acidosis and hyperventilation. Maintained homeostasis mitigates the effects of shunt. PMID- 18423017 TI - PCR-DGGE assessment of the bacterial diversity of breast milk in women with lactational infectious mastitis. AB - BACKGROUND: Infectious mastitis is a common condition during lactation and in fact, represents one of the main causes leading to a precocious weaning. The number of studies dealing with lactational mastitis is low and, up to now, the etiological diagnosis is frequently made on the basis of unspecific clinical signs. The aim of this study was to investigate the microbial diversity of breast milk in 20 women with lactational mastitis employing culture-dependent and culture-independent (PCR-DGGE) approaches. METHODS: Breast milk samples were cultured in different media to investigate the presence of bacteria and/or yeasts, and a total of 149 representative isolates were identified to the species level by 16S rRNA gene PCR sequencing. The microorganisms recovered were compared with those found by PCR-DGGE analysis. To identify the DGGE profiles two reference markers of different microbial species were constructed. Sequence analysis of unknown bands was also performed. RESULTS: Staphylococci were the dominant bacterial group and Staphylococcus epidermidis was the dominant species. In a lower number of samples, other bacteria (mainly streptococci and a few gram negative species) were also identified. Globally, PCR-DGGE results showed a good correlation with those obtained by culture-based methods. However, although DNA bands corresponding to different lactic acid bacteria were detected, such bacteria could not be isolated from the milk samples. CONCLUSION: Staphylococci seem to be the main etiological agents of human lactational mastitis. The combined use of culture and molecular techniques allowed a better characterization of the bacterial diversity in milk from women suffering from infectious mastitis. Our results suggest that this condition could be the result of a disbiotic process where some of the bacterial species usually present in human milk outgrow (staphylococci) while others disappear (lactobacilli or lactococci). PMID- 18423018 TI - Mosquito abundance, bed net coverage and other factors associated with variations in sporozoite infectivity rates in four villages of rural Tanzania. AB - BACKGROUND: Entomological surveys are of great importance in decision-making processes regarding malaria control strategies because they help to identify associations between vector abundance both species-specific ecology and disease intervention factors associated with malaria transmission. Sporozoite infectivity rates, mosquito host blood meal source, bed net coverage and mosquito abundance were assessed in this study. METHODOLOGY: A longitudinal survey was conducted in four villages in two regions of Tanzania. Malaria vectors were sampled using the CDC light trap and pyrethrum spray catch methods. In each village, ten paired houses were selected for mosquitoes sampling. Sampling was done in fortnight case and study was undertaken for six months in both Kilimanjaro (Northern Tanzania) and Dodoma (Central Tanzania) regions. RESULTS: A total of 6,883 mosquitoes were collected including: 5,628 (81.8%) Anopheles arabiensis, 1,100 (15.9%) Culex quinquefasciatus, 89 (1.4%) Anopheles funestus, and 66 (0.9%) Anopheles gambiae s.s. Of the total mosquitoes collected 3,861 were captured by CDC light trap and 3,022 by the pyrethrum spray catch method. The overall light trap: spray catch ratio was 1.3:1. Mosquito densities per room were 96.5 and 75.5 for light trap and pyrethrum spray catch respectively. Mosquito infectivity rates between villages that have high proportion of bed net owners and those without bed nets was significant (P < 0.001) and there was a significant difference in sporozoite rates between households with and without bed nets in these four villages (P < 0.001). CONCLUSION: Malaria remains a major problem in the study areas characterized as low transmission sites. Further studies are required to establish the annual entomological inoculation rates and to observe the annual parasitaemia dynamics in these communities. Outdoor mosquitoes collection should also be considered. PMID- 18423019 TI - Alterations in plasma soluble vascular endothelial growth factor receptor-1 (sFlt 1) concentrations during coronary artery bypass graft surgery: relationships with post-operative complications. AB - BACKGROUND: Plasma concentrations of sFlt-1, the soluble form of the vascular endothelial growth factor receptor (VEGF), markedly increase during coronary artery bypass graft (CABG) surgery with extracorporeal circulation (ECC). We investigated if plasma sFlt-1 values might be related to the occurrence of surgical complications after CABG. METHODS: Plasma samples were collected from the radial artery catheter before vascular cannulation and after opening the chest, at the end of ECC just before clamp release, after cross release, after weaning from ECC, at the 6th and 24th post-operative hour. Thirty one patients were investigated. The presence of cardiovascular, haematological and respiratory dysfunctions was prospectively assessed. Plasma sFlt-1 levels were measured with commercially ELISA kits. RESULTS: Among the 31 investigated patients, 15 had uneventful surgery. Patients with and without complications had similar pre operative plasma sFlt-1 levels. Lowered plasma sFlt-1 levels were observed at the end of ECC in patients with haematological (p = 0.001, ANOVA) or cardiovascular (p = 0.006) impairments, but not with respiratory ones (p = 0.053), as compared to patients with uneventful surgery. CONCLUSION: These results identify an association between specific post-CABG complication and the lower release of sFlt 1 during ECC. sFlt-1-induced VEGF neutralisation might, thus, be beneficial to reduce the development of post-operative adverse effects after CABG. PMID- 18423021 TI - Endorsement of the CONSORT Statement by high impact factor medical journals: a survey of journal editors and journal 'Instructions to Authors'. AB - BACKGROUND: The CONSORT Statement provides recommendations for reporting randomized controlled trials. We assessed the extent to which leading medical journals that publish reports of randomized trials incorporate the CONSORT recommendations into their journal and editorial processes. METHODS: This article reports on two observational studies. Study 1: We examined the online version of 'Instructions to Authors' for 165 high impact factor medical journals and extracted all text mentioning the CONSORT Statement or CONSORT extension papers. Any mention of the International Committee of Medical Journal Editors (ICMJE) or clinical trial registration were also sought and extracted. Study 2: We surveyed the editor-in-chief, or editorial office, for each of the 165 journals about their journal's endorsement of CONSORT recommendations and its incorporation into their editorial and peer-review processes. RESULTS: Study 1: Thirty-eight percent (62/165) of journals mentioned the CONSORT Statement in their online 'Instructions to Authors'; of these 37% (23/62) stated this was a requirement, 63% (39/62) were less clear in their recommendations. Very few journals mentioned the CONSORT extension papers. Journals that referred to CONSORT were more likely to refer to ICMJE guidelines (RR 2.16; 95% CI 1.51 to 3.08) and clinical trial registration (RR 3.67; 95% CI 2.36 to 5.71) than those journals which did not.Study 2: Thirty-nine percent (64/165) of journals responded to the on-line survey, the majority were journal editors. Eighty-eight percent (50/57) of journals recommended authors comply with the CONSORT Statement; 62% (35/56) said they would require this. Forty-one percent (22/53) reported incorporating CONSORT into their peer-review process and 47% (25/53) into their editorial process. Eighty-one percent (47/58) reported including CONSORT in their 'Instructions to Authors' although there was some inconsistency when cross checking information on the journal's website. Sixty-nine percent (31/45) of journals recommended authors comply with the CONSORT extension for cluster trials, 60% (27/45) for harms and 42% (19/45) for non-inferiority and equivalence trials. Few journals mentioned these extensions in their 'Instructions to Authors'. CONCLUSION: Journals should be more explicit in their recommendations and expectations of authors regarding the CONSORT Statement and related CONSORT extensions papers. PMID- 18423020 TI - Overexpression of mtDNA-associated AtWhy2 compromises mitochondrial function. AB - BACKGROUND: StWhy1, a member of the plant-specific Whirly single-stranded DNA binding protein family, was first characterized as a transcription factor involved in the activation of the nuclear PR-10a gene following defense-related stress in potato. In Arabidopsis thaliana, Whirlies have recently been shown to be primarily localized in organelles. Two representatives of the family, AtWhy1 and AtWhy3 are imported into plastids while AtWhy2 localizes to mitochondria. Their function in organelles is currently unknown. RESULTS: To understand the role of mitochondrial Whirlies in higher plants, we produced A. thaliana lines with altered expression of the atwhy2 gene. Organellar DNA immunoprecipitation experiments demonstrated that AtWhy2 binds to mitochondrial DNA. Overexpression of atwhy2 in plants perturbs mitochondrial function by causing a diminution in transcript levels and mtDNA content which translates into a low activity level of respiratory chain complexes containing mtDNA-encoded subunits. This lowered activity of mitochondria yielded plants that were reduced in size and had distorted leaves that exhibited accelerated senescence. Overexpression of atwhy2 also led to early accumulation of senescence marker transcripts in mature leaves. Inactivation of the atwhy2 gene did not affect plant development and had no detectable effect on mitochondrial morphology, activity of respiratory chain complexes, transcription or the amount of mtDNA present. This lack of phenotype upon abrogation of atwhy2 expression suggests the presence of functional homologues of the Whirlies or the activation of compensating mechanisms in mitochondria. CONCLUSION: AtWhy2 is associated with mtDNA and its overexpression results in the production of dysfunctional mitochondria. This report constitutes the first evidence of a function for the Whirlies in organelles. We propose that they could play a role in the regulation of the gene expression machinery of organelles. PMID- 18423022 TI - Missing value imputation improves clustering and interpretation of gene expression microarray data. AB - BACKGROUND: Missing values frequently pose problems in gene expression microarray experiments as they can hinder downstream analysis of the datasets. While several missing value imputation approaches are available to the microarray users and new ones are constantly being developed, there is no general consensus on how to choose between the different methods since their performance seems to vary drastically depending on the dataset being used. RESULTS: We show that this discrepancy can mostly be attributed to the way in which imputation methods have traditionally been developed and evaluated. By comparing a number of advanced imputation methods on recent microarray datasets, we show that even when there are marked differences in the measurement-level imputation accuracies across the datasets, these differences become negligible when the methods are evaluated in terms of how well they can reproduce the original gene clusters or their biological interpretations. Regardless of the evaluation approach, however, imputation always gave better results than ignoring missing data points or replacing them with zeros or average values, emphasizing the continued importance of using more advanced imputation methods. CONCLUSION: The results demonstrate that, while missing values are still severely complicating microarray data analysis, their impact on the discovery of biologically meaningful gene groups can - up to a certain degree - be reduced by using readily available and relatively fast imputation methods, such as the Bayesian Principal Components Algorithm (BPCA). PMID- 18423023 TI - Dysregulated angiogenesis in B-chronic lymphocytic leukemia: morphologic, immunohistochemical, and flow cytometric evidence. AB - BACKGROUND: The extent of enhanced bone marrow angiogenesis in chronic lymphocytic leukemia (CLL) and relationship to proangiogenic factors and prognostic indicators is largely unexplored. METHODS: To further investigate the role of angiogenesis in CLL by evaluating the topography and extent of angiogenesis in a group of CLL bone marrow biopsies, to study the expression of pro and antiangiogenic vascular factors in CLL cells to more precisely document the cell types producing these factors, and to evaluate the role, if any, of localized hypoxia in upregulation of angiogenesis in CLL We used immunohistochemistry (IHC) (n = 21 pts) with antibodies to CD3 and CD20, proangiogenic (VEGF, HIF-1a) and antiangiogenic (TSP-1) factors, and VEGF receptors -1 and -2 to examine pattern/extent of CLL marrow involvement, microvessel density (MVD), and angiogenic characteristics; flow cytometry (FC) was performed on 21 additional cases for VEGF and TSP-1. RESULTS: CLL patients had higher MVD (23.8 vs 14.6, p~0.0002) compared to controls (n = 10). MVD was highest at the periphery of focal infiltrates, was not enhanced in proliferation centers, and was increased irrespective of the presence or absence of cytogenetic/immunophenotypic markers of aggressivity. By IHC, CLL cells were VEGF(+), HIF-1a (+), TSP-1(-), VEGFR-1(+), and VEGFR-2(+). By FC, CLL cells were 1.4-2.0-fold brighter for VEGF than T cells and were TSP-1(-). CONCLUSION: CLL demonstrates enhanced angiogenesis, with increased MVD, upregulated VEGF and downregulated TSP-1. Upregulation of HIF-1a in all CLL cases suggests localized tissue hypoxia as an important stimulant of microvessel proliferation. The presence of VEGF receptors on CLL cells implies an autocrine effect for VEGF. Differences in MVD did not correlate with traditional genetic/immunophenotypic markers of aggressiveness. PMID- 18423024 TI - Diet and cardiovascular risk in university marching band, dance team and cheer squad members: a cross-sectional study. AB - BACKGROUND: Cardiovascular disease (CVD) is the leading cause of death in the United States. Diets high in fat, especially saturated fat, are often linked to obesity, hypertension and hypercholesterolemia, all risk factors for CVD. The purpose of this study was to determine the association between diet and CVD risk factors in members of a university marching band, dance team and cheer squad. METHODS: In 2004, 232 marching band, dance team and cheer squad members completed a self-administered survey evaluating dietary intake. Body mass index (BMI), waist-to-hip ratio (WHR), blood pressure, fasting serum glucose and cholesterol were measured. Unpaired t-test and Pearson's chi square test were used to determine baseline differences by gender. Multiple linear regression analysis was used to determine the cross-sectional association between dietary intake of various food groups such as grains, meats, fruits & vegetables, dairy, water, alcohol and risk factors for CVD namely BMI, WHR, blood glucose, total cholesterol, and blood pressure (BP). RESULTS: 45% of the participants were overweight; 30% of females and 4.3% of males had WHR >/= 0.80 and 0.95 respectively. Almost 8% were hyperglycemic, 10% hypercholesterolemic, 15% had high systolic and 9% had high diastolic BP. Less than 50% consumed the recommended servings of grains, fruits and vegetables, dairy and water and 58% consumed alcohol. Higher grains intake was positively associated with higher BMI (Adjusted beta = 1.97, p = 0.030, 95% CI: 0.19, 3.74) and; higher alcohol intake was also positively associated with higher BMI (Adjusted beta = 0.15, p = 0.002, 95% CI: 0.06, 0.24). CONCLUSION: These results warrant the evaluation of existing college-based health programs and development of new interventions to improve dietary habits and promote a healthy lifestyle in these athletes. PMID- 18423026 TI - Why do people fail to turn good intentions into action? The role of executive control processes in the translation of healthy eating intentions into action in young Scottish adults. AB - BACKGROUND: Despite the significant health benefits associated with eating healthily, diet is extremely difficult to change, with the majority of people who intend to eat more healthily failing to do so. Recent evidence has suggested that the ability to turn intentions into actions may be related to individual differences in one facet of executive control - cognitive inhibition (i.e. the ability to inhibit irrelevant information and suppress prepotent responses). The present study investigates the role of this and other executive processes (inhibition, task switching, planning and cognitive flexibility) in the translation of dietary intentions into action. In addition, as the literature suggests that weak executive control may be associated with hyper-responsivity to cues to action, the role of executive processes in susceptibility to environmental food cues and responses to If-Then plans designed to cue intended behaviour are investigated. METHODS: Future intentions about consumption of fruits and vegetables and snack foods will be measured in a sample of young adults. Actual consumption of the target foods will be recorded with computerised diaries over a subsequent 3-day period. Performance on a battery of established executive control tasks (Go-NoGo, Tower task, Verbal Fluency task and Trail Making) will be used to predict the discrepancy between intended and actual dietary behaviour. In addition, executive control scores will be used to predict reported susceptibility to environmental food cues and benefit derived from the use of 'If-Then plans' designed to cue intended behaviour. DISCUSSION: Our findings will add to understanding about the role of executive control in translating intentions into actions and may demonstrate potential for future public health interventions. If participants with weak executive control are found to be less likely to eat as they intend than those with strong executive control, then interventions that reduce the load on these executive processes may increase chances of successful intention-behaviour translation. If those with weak executive control are found to be more responsive to cues to action they may also benefit more from the use of If-Then plans designed to cue intended behaviour. PMID- 18423025 TI - Socio-economic status is inversely related to bed net use in Gabon. AB - BACKGROUND: Insecticide-treated bed nets (ITNs) range among the most effective measures of malaria prophylaxis, yet their implementation level in sub-Saharan Africa is still low. The goal of this study was to investigate the influence of socio-economic factors on the use of bed nets by mothers in Gabon. METHODS: A cross-sectional study was conducted completing pre-tested, interviewer administered questionnaires exploring socioeconomic proxy measures with 397 mothers or guardians of young children. Respondents were grouped according to their socio-economic situation, using scores. The condition of the bed nets was evaluated during a home visit. RESULTS: Socio-economic factors of wellbeing were negatively associated with bed net use, such as living in a stone house (OR 0.26, 95% CI 0.14-0.48), running water in the house (OR 0.44, 95% CI 0.21-0.92), shower/flush toilet in the house (OR 0.39/0.34, 95% CI 0.21-0.75/0.16-0.73), ownership of a freezer (OR 0.50, 95% CI 0.26-0.96) and belonging to the highest group in the economic score (OR 0.32, 95% CI 0.15-0.67). In contrast, similar factors were positively associated with a good maintenance condition of the bed nets: higher monthly income (OR 5.64, 95% CI 2.41-13.19) and belonging to the highest group in the economic score (OR 2.55, 95% CI 1.19 - 5.45). CONCLUSION: Among the poorest families in Lambarene the coverage with untreated nets (UTNs) is the highest, but the condition of these UTNs is the worst. To achieve a broad implementation of ITNs in Lambarene, there is an urgent need for educational programmes as well as need-tailored marketing strategies for ITNs. PMID- 18423027 TI - Surgical treatment of scoliosis: a review of techniques currently applied. AB - In this review, basic knowledge and recent innovation of surgical treatment for scoliosis will be described. Surgical treatment for scoliosis is indicated, in general, for the curve exceeding 45 or 50 degrees by the Cobb's method on the ground that:1) Curves larger than 50 degrees progress even after skeletal maturity.2) Curves of greater magnitude cause loss of pulmonary function, and much larger curves cause respiratory failure.3) Larger the curve progress, more difficult to treat with surgery.Posterior fusion with instrumentation has been a standard of the surgical treatment for scoliosis. In modern instrumentation systems, more anchors are used to connect the rod and the spine, resulting in better correction and less frequent implant failures. Segmental pedicle screw constructs or hybrid constructs using pedicle screws, hooks, and wires are the trend of today.Anterior instrumentation surgery had been a choice of treatment for the thoracolumbar and lumbar scoliosis because better correction can be obtained with shorter fusion levels. Recently, superiority of anterior surgery for the thoracolumbar and lumbar scoliosis has been lost. Initial enthusiasm for anterior instrumentation for the thoracic curve using video assisted thoracoscopic surgery technique has faded out.Various attempts are being made with use of fusionless surgery. To control growth, epiphysiodesis on the convex side of the deformity with or without instrumentation is a technique to provide gradual progressive correction and to arrest the deterioration of the curves. To avoid fusion for skeletally immature children with spinal cord injury or myelodysplasia, vertebral wedge ostetomies are performed for the treatment of progressive paralytic scoliosis. For right thoracic curve with idiopathic scoliosis, multiple vertebral wedge osteotomies without fusion are performed. To provide correction and maintain it during the growing years while allowing spinal growth for early onset scoliosis, technique of instrumentation without fusion or with limited fusion using dual rod instrumentation has been developed. To increase the volume of the thorax in thoracic insufficiency syndrome associated with fused ribs and congenital scoliosis, vertical expandable prosthetic titanium ribs has been developed. PMID- 18423028 TI - Review of the utilization of HEEPF--competitive projects for educational enhancement in the Egyptian medical sector. AB - In Egypt, the medical sector has been facing the same problems that challenged the system of higher education in the past decades, mainly an increasing student enrollment, limited resources, and old governance and bylaws. These constraints and the escalating paucity of resources have had a major negative influence on quality of education. Consequently, thoughts of educational reform came forward in the form of competitive projects, which have attracted several institutes from the health sector to improve their educational performance. The aim of this paper is to review the share of the medical sector in the higher education enhancement project fund (HEEPF), its outcomes, sustainability, and to provide recommendations for keeping the momentum of reform pursuit in the future. The methodology included obtaining statistics pertaining to the medical sector in Egypt as regards colleges, students, and staff. We also reviewed the self-studies of the medical sector colleges, HEEPF projects reports, performance appraisal reports, and World Bank reports on HEEPF achievements in order to retrieve the required data. Results showed that medical sector had a large share of the HEEPF (28.5% of projects) as compared to its size (8% of student population). The projects covered 10 areas; the frequency distribution of which ranged between 4.4% (creation of new programs) to 97.8% (human resource development). In conclusion, educational enhancement in the medical sector in Egypt could be apparently achieved through the HEEPF competitive projects. A study of the long term impact of these projects on the quality of education is recommended. PMID- 18423029 TI - Validation of an individualised quality of life measure in older day hospital patients. AB - BACKGROUND: To test the ease of use, reliability, responsiveness and construct validity of the Patient Generated Index, an individualised quality of life score, in older people attending a Medicine for Older People Day Hospital. METHODS: Prospective longitudinal study in patients attending a specialist Medicine for Older People Day Hospital in Scotland. The Patient Generated Index was administered at baseline, one week later, and at the end of Day Hospital attendance. Functional Limitations Profile, Hospital Anxiety and Depression Score, Barthel index and global subjective impressions of change were also collected and compared with baseline scores and change in Patient Generated Index scores. Reliability was assessed using intraclass correlation coefficients in subjects reporting no change in global quality of life; responsiveness was assessed using effect size and Guyatt coefficients in subjects reporting change in global quality of life. External validity was assessed via correlation with measures of physical function, comorbid disease and psychological state. RESULTS: 75 patients were enrolled, mean age 81 years. Mean completion time was 5.0 minutes at baseline. Reliability was moderate (intraclass correlation coefficient 0.72) but there were weak and inconsistent responses to change (effect sizes 0.02 to 0.15; Guyatt responsiveness coefficient 0.29). Patient Generated Index scores correlated with Functional Limitation Profile scores (r = 0.51, p < 0.001), baseline anxiety score (r = -0.25, P = 0.039) and baseline depression score (r = 0.37, P = 0.002) but displayed only weak, non-significant correlation with number of comorbid diseases (r = -0.22, P = 0.07), number of medications (r = -0.21, P = 0.08) and Barthel score (r = 0.09, p = 0.45). CONCLUSION: The Patient Generated Index appears moderately reliable and easy to complete, but is poorly responsive to change, limiting its usefulness in clinical practice or research. PMID- 18423030 TI - Expanded risk groups help determine which prostate radiotherapy sub-group may benefit from adjuvant androgen deprivation therapy. AB - PURPOSE: To assess whether an expanded (five level) risk stratification system can be used to identify the sub-group of intermediate risk patients with prostate cancer who benefit from combining androgen deprivation therapy (ADT) with external beam radiotherapy (EBRT). MATERIALS AND METHODS: Using a previously validated 5-risk group schema, a prospective non-randomized data set of 1423 men treated at the British Columbia Cancer Agency was assessed for the primary end point of biochemical control (bNED) with the RTOG-ASTRO "Phoenix" definition (lowest PSA to date + 2 ng/mL), both with and without adjuvant ADT. The median follow-up was 5 years. RESULTS: There was no bNED benefit for ADT in the low or low intermediate groups but there was a statistically significant bNED benefit in the high intermediate, high and extreme risk groups. The 5-year bNED rates with and without ADT were 70% and 73% respectively for the low intermediate group (p = non-significant) and 72% and 58% respectively for the high intermediate group (p = 0.002). CONCLUSION: There appears to be no advantage to ADT where the Gleason score is 6 or less and PSA is 15 or less. ADT is beneficial in patients treated to standard dose radiation with Gleason 6 disease and a PSA greater than 15 or where the Gleason score is 7 or higher. PMID- 18423031 TI - Image standards in tissue-based diagnosis (diagnostic surgical pathology). AB - BACKGROUND: Progress in automated image analysis, virtual microscopy, hospital information systems, and interdisciplinary data exchange require image standards to be applied in tissue-based diagnosis. AIMS: To describe the theoretical background, practical experiences and comparable solutions in other medical fields to promote image standards applicable for diagnostic pathology. THEORY AND EXPERIENCES: Images used in tissue-based diagnosis present with pathology specific characteristics. It seems appropriate to discuss their characteristics and potential standardization in relation to the levels of hierarchy in which they appear. All levels can be divided into legal, medical, and technological properties. Standards applied to the first level include regulations or aims to be fulfilled. In legal properties, they have to regulate features of privacy, image documentation, transmission, and presentation; in medical properties, features of disease-image combination, human-diagnostics, automated information extraction, archive retrieval and access; and in technological properties features of image acquisition, display, formats, transfer speed, safety, and system dynamics. The next lower second level has to implement the prescriptions of the upper one, i.e. describe how they are implemented. Legal aspects should demand secure encryption for privacy of all patient related data, image archives that include all images used for diagnostics for a period of 10 years at minimum, accurate annotations of dates and viewing, and precise hardware and software information. Medical aspects should demand standardized patients' files such as DICOM 3 or HL 7 including history and previous examinations, information of image display hardware and software, of image resolution and fields of view, of relation between sizes of biological objects and image sizes, and of access to archives and retrieval. Technological aspects should deal with image acquisition systems (resolution, colour temperature, focus, brightness, and quality evaluation procedures), display resolution data, implemented image formats, storage, cycle frequency, backup procedures, operation system, and external system accessibility. The lowest third level describes the permitted limits and threshold in detail. At present, an applicable standard including all mentioned features does not exist to our knowledge; some aspects can be taken from radiological standards (PACS, DICOM 3); others require specific solutions or are not covered yet. CONCLUSION: The progress in virtual microscopy and application of artificial intelligence (AI) in tissue-based diagnosis demands fast preparation and implementation of an internationally acceptable standard. The described hierarchic order as well as analytic investigation in all potentially necessary aspects and details offers an appropriate tool to specifically determine standardized requirements. PMID- 18423032 TI - PRRDB: a comprehensive database of pattern-recognition receptors and their ligands. AB - BACKGROUND: Recently in a number of studies, it has been demonstrated that the innate immune system doesn't merely acts as the first line of defense but provides critical signals for the development of specific adaptive immune response. Innate immune system employs a set of receptors called pattern recognition receptors (PRRs) that recognize evolutionarily conserved patterns from pathogens called pathogen associated molecular patterns (PAMPs). In order to assist scientific community, a database PRRDB has been developed that provides extensive information about pattern recognition receptors and their ligands. RESULTS: The current version of database contains around 500 patterns recognizing receptors from 77 distinct organisms ranging from insects to human. This includes 177 Toll-like receptors, 124 are Scavenger receptors and 67 are Nucleotide Binding Site-Leucine repeats rich receptors. The database also provides information about 266 ligands that includes carbohydrates, proteins, nucleic acids, glycolipids, glycoproteins, lipopeptides. A number of web tools have been integrated in PRRDB in order to provide following services: i) searching on any field; ii) database browsing; and iii) BLAST search against the pattern recognition receptors. PRRDB also provides external links to standard databases like Swiss-Prot and Pubmed. CONCLUSION: PRRDB is a unique database of its kind, which provides comprehensive information about innate immunity. This database will be very useful in designing effective adjuvant for subunit vaccine and in understanding role of innate immunity. The database is available from the URL's in the Availabiltiy and requirements section. PMID- 18423034 TI - The versatile reverse flow sural artery neurocutaneous flap: a case series and review of literature. AB - BACKGROUND: Reverse flow sural neurocutaneous flap has been utilized more frequently during the past decade to cover vital structures around the foot and ankle area. The potential advantages are the relatively constant blood supply, ease of elevation and preservation of major vascular trunks in the leg. The potential disadvantages remain venous congestion, donor site morbidity and lack of sensation. METHODS: This descriptive case series was conducted at Queen Mary Hospital, Hong Kong, from 1997 to 2003. Ten patients having undergone reverse flow sural neurocutaneous flap were identified through medical records. There were six females (60%) and four males (40%), with an average age of 59.8 years. The defects occurred as a result of trauma in five patients (50%), diabetic ulcers in four (40%) and decubitus ulcer in one (10%) paraplegic patient. The defect site included non weight bearing heel in four (40%), tendo Achilles in two (20%), distal tibia in two (20%), lateral malleolus in one (10%) and medial aspect of the midfoot in one patient (10%). The maximum flap size harvested was 14 x 6 cm. Preoperative doppler evaluation was performed in all patients to identify perforators and modified plaster of paris boot was used in the post operative period. A detailed questionnaire was developed addressing variables of interest. RESULTS: There was no flap failure. Venous congestion was encountered in one case. The donor site was relatively unsightly but acceptable to all patients. The loss of sensation in the sural nerve distribution was transient in all patients. CONCLUSION: Reverse sural artery flap remains to be the workhorse flap to resurface the soft tissue defects of the foot and ankle. Anastomosis of the sural nerve to the digital plantar nerve can potentially solve the issue of lack of sensation in the flap especially when used for weight bearing heel. PMID- 18423033 TI - Survival time in severe hemorrhagic shock after perioperative hemodilution is longer with PEG-conjugated human serum albumin than with HES 130/0.4: a microvascular perspective. AB - INTRODUCTION: Preoperative hemodilution is an established practice that is applied to reduce surgical blood loss. It has been proposed that polyethylene glycol (PEG) surface decorated proteins such as PEG-conjugated human serum albumin may be used as non-oxygen-carrying plasma expanders. The purpose of this study was to determine whether there is any difference in survival time after severe hemorrhagic shock following extreme hemodilution using a conventional hydroxyethyl starch (HES)-based plasma expander or PEG-albumin. METHODS: Experiments were performed using the hamster skinfold window preparation. Human serum albumin that was surface decorated with PEG was compared with Voluven 6% (Fresenius Kabi, Austria; a starch solution that is of low molecular weight and has a low degree of substitution; HES). These plasma expanders were used for a 50% (blood volume) exchange transfusion to simulate preoperative hemodilution. Exchange transfusion was followed by a 60% (blood volume) hemorrhage to reproduce a severe surgical bleed over a 1 hour period. Observation of the animal was continued for another hour during the shock phase. RESULTS: The PEG-albumin group exhibited significantly greater survival rate than did the HES group, in which none of the animals survived the hemorrhage phase of the experiment. Among the treatment groups there were no changes in mean arterial pressure and heart rate from baseline after hemodilution. Both groups experienced gradual increases in arterial oxygen tension and disturbance in acid-base balance, but this response was more pronounced in the HES group during the shock period. Mean arterial pressure remained elevated after the initial hemorrhage period in the PEG-albumin group but not in the HES group. Maintenance of a greater mean arterial pressure during the initial stages of hemorrhage is proposed to be in part due to the improved volume expansion with PEG-albumin, as indicated by the significant decrease in systemic hematocrit compared with the HES group. PEG-albumin treatment yielded higher functional capillary density during the initial stages of hemorrhage as compared with HES treatment. CONCLUSION: The ability of PEG albumin to prolong maintenance of microvascular function better than HES is a finding that would be significant in a clinical setting involving preoperative blood management and extreme blood loss. PMID- 18423035 TI - Do airway metallic stents for benign lesions confer too costly a benefit? AB - BACKGROUND: The use of self-expanding metallic stents (SEMAS) in the treatment benign airway obstruction is controversial. METHODS: To evaluate the safety and efficacy of SEMAS for this indication, we conducted a 10-year retrospective review at our tertiary medical centre. RESULTS: Using flexible bronchoscopy, 82 SEMAS (67% Ultraflex, 33% Wallstent) were placed in 35 patients with inoperable lesions, many with significant medical comorbidities (88%). 68% of stents were tracheal, and 83% of patients showed immediate symptomatic improvement. Reversible complications developed in 9% of patients within 24 hrs of stent placement. Late complications (>24 hrs) occurred in 77% of patients, of which 37% were clinically significant or required an interventional procedure. These were mainly due to stent migration (12.2%), fracture (19.5%), or obstructive granulomas (24.4%). The overall granuloma rate of 57% was higher at tracheal sites (59%) than bronchial ones (34%), but not significantly different between Ultraflex and Wallstents. Nevertheless, Wallstents were associated with higher rates of bleeding (5% vs. 30%, p = 0.005) and migration (7% vs. 26%, p = 0.026). Of 10 SEMAS removed using flexible bronchoscopy, only one was associated with incomplete removal of fractured stent wire. Median survival was 3.6 +/- 2.7 years. CONCLUSION: Ill patients with inoperable lesions may be considered for treatment with SEMAS. PMID- 18423036 TI - Parents' concerns about children are highly prevalent but often not confirmed by child doctors and nurses. AB - BACKGROUND: The aim of this study was to assess the prevalence in the general population of parents' concerns about the development of their child, to identify groups at risk and to assess the association between parents' concerns and professional judgement. METHODS: We obtained cross-sectional data on a Dutch nationally representative sample of children aged 14 months, 3 3/4, 5-6 and 8-12 years within the setting of routine well-child visits provided to the entire population. A total of 4,107 participated (response rate 85.3%). Data were about concerns that parents reported by questionnaire before the visit regarding behavioural and emotional problems, developmental delay, consequences of disease and contact with peers that needed professional assistance, and about the assessment of these domains by doctors and nurses during the visit. Moreover, we obtained data on parent-reported psychosocial problems using the Infant-Toddler Social and Emotional Assessment and the Child Behavior Checklist. RESULTS: Of all parents, 49.3% reported some concerns and 8.7% reported frequent concerns, most frequently on child behaviour. Frequent concerns were most likely to refer to young children, children from labour immigrant families, with fathers of medium educational level and in low-income families. The prevalence rates of professional-assessed parenting problems were much lower than parent-reported ones. The rates of psychosocial problems were highest in the case of shared concerns, but also higher if parents expressed concerns that were not confirmed by professionals. CONCLUSION: A very large proportion of parents of young children have concerns regarding their child, but agreement on these concerns with child health professionals is relatively low. PMID- 18423037 TI - Characterization of a panel of six beta2-adrenergic receptor antibodies by indirect immunofluorescence microscopy. AB - BACKGROUND: The beta2-adrenergic receptor (beta2AR) is a primary target for medications used to treat asthma. Due to the low abundance of beta2AR, very few studies have reported its localization in tissues. However, the intracellular location of beta2AR in lung tissue, especially in airway smooth muscle cells, is very likely to have a significant impact on how the airways respond to beta agonist medications. Thus, a method for visualizing beta2AR in tissues would be of utility. The purpose of this study was to develop an immunofluorescent labeling technique for localizing native and recombinant beta2AR in primary cell cultures. METHODS: A panel of six different antibodies were evaluated in indirect immunofluorescence assays for their ability to recognize human and rat beta2AR expressed in HEK 293 cells. Antibodies capable of recognizing rat beta2AR were identified and used to localize native beta2AR in primary cultures of rat airway smooth muscle and epithelial cells. beta2AR expression was confirmed by performing ligand binding assays using the beta-adrenergic antagonist [3H] dihydroalprenolol ([3H]DHA). RESULTS: Among the six antibodies tested, we identified three of interest. An antibody developed against the C-terminal 15 amino acids of the human beta2AR (Ab-Bethyl) specifically recognized human but not rat beta2AR. An antibody developed against the C-terminal domain of the mouse beta2AR (Ab-sc570) specifically recognized rat but not human beta2AR. An antibody developed against 78 amino acids of the C-terminus of the human beta2AR (Ab 13989) was capable of recognizing both rat and human beta2ARs. In HEK 293 cells, the receptors were predominantly localized to the cell surface. By contrast, about half of the native rat beta2AR that we visualized in primary cultures of rat airway epithelial and smooth muscle cells using Ab-sc570 and Ab-13989 was found inside cells rather than on their surface. CONCLUSION: Antibodies have been identified that recognize human beta2AR, rat beta2AR or both rat and human beta2AR. Interestingly, the pattern of expression in transfected cells expressing millions of receptors was dramatically different from that in primary cell cultures expressing only a few thousand native receptors. We anticipate that these antibodies will provide a valuable tool for evaluating the expression and trafficking of beta2AR in tissues. PMID- 18423038 TI - Late pulmonary metastases of renal cell carcinoma immediately after post transplantation immunosuppressive treatment: a case report. AB - INTRODUCTION: We report a case of pulmonary metastatic recurrence of renal adenocarcinoma soon after radical nephrectomy that was followed by renal transplant and immunosuppressive medication. Increased risk of metastatic recurrence of renal cell carcinoma should be considered in the immediate post transplant period when immunosuppressive medication is administered, even if nephrectomy had been performed many years earlier. CASE PRESENTATION: In 1986 the patient demonstrated renal insufficiency secondary to mesangial glomerulonephritis. In 1992 he underwent left side radical nephrectomy with histopathological diagnosis of clear cell adenocarcinoma. Mesangial glomerulonephritis in the remaining right kidney progressed to end-stage renal failure. In October 2000 he received a kidney transplant from a cadaver and commenced immunosuppressive medication. Two months later, several nodules were found in his lungs, which were identified as metastases from the primary renal tumor that had been removed with the diseased kidney 8 years earlier. CONCLUSION: Recurrence of renal cell carcinoma metastases points to tumor dormancy and reflects a misbalance between effective tumor immune surveillance and immune escape. This case demonstrates that a state of tumor dormancy can be interrupted soon after administration of immunosuppressant medication. PMID- 18423039 TI - In vivo endothelial gene regulation in diabetes. AB - BACKGROUND: An authentic survey of the transcript-level response of the diabetic endothelium in vivo is key to understanding diabetic cardiovascular complications such as accelerated atherosclerosis and endothelial dysfunction. METHODS: We used streptozotocin to induce a model of type I diabetes in transgenic mice that express green fluorescent protein under the control of an endothelial-specific promoter (Tie2-GFP) allowing rapid isolation of aortic endothelium. Three weeks after treatment, endothelial cells were isolated from animals with blood glucose > 350 mg/dl. Aortae from the root to the renal bifurcation were rapidly processed by mincing and proteolytic digestion followed by fluorescent activated cell sorting to yield endothelial cell populations of >95% purity. RNA was isolated from >50,000 endothelial cells and subjected to oligo dT amplification prior to transcriptional analysis on microarrays displaying long oligonucleotides representing 32,000 murine transcripts. Five regulated transcripts were selected for analysis by real-time PCR. RESULTS: Within replicate microarray experiments, 19 transcripts were apparently dysregulated by at least 70% within diabetic mice. Up-regulation of glycam1, slc36a2, ces3, adipsin and adiponectin was confirmed by real-time PCR. CONCLUSION: By comprehensively examining cellular gene responses in vivo in a whole animal model of type I diabetes, we have identified novel regulation of key endothelial transcripts that likely contribute to the metabolic and pro-inflammatory responses that accompany diabetes. PMID- 18423041 TI - Correlates of low birth weight in term pregnancies: a retrospective study from Iran. AB - BACKGROUND: Low birth weight (LBW) is considered as a major multifaceted public health concern. Seventy-two percent of LBW infants are born in Asia. An estimation of 8% LBW infants has been reported for Eastern Mediterranean region including Iran. This study investigated contributory factors of LBW in singleton term births in Tehran, Iran. Tehran is a multicultural metropolitan area and a sample from the general population in Tehran could be regarded as a representative sample of urban population in Iran. METHODS: This was a retrospective study using data from 15 university maternity hospitals in Tehran, Iran. Data on all singleton term births in these hospitals were extracted from case records during a one calendar year. Study variables included: maternal age, maternal educational level, history of LBW deliveries, history of preterm labor, cigarette smoking during pregnancy, number of parities, chronic diseases and residential area (Tehran versus suburbs of Tehran). In order to examine the relationship between LBW and demographic and reproductive variables the adjusted logistic regression analysis was performed. RESULTS: In all, data for 3734 term pregnancies were extracted. The mean age of women was 25.7 (SD = 5.3) years and 5.2% of term births were LBW. In addition to association between LBW and maternal age, significant risk factors for LBW were: history of LBW deliveries [adjusted odds ratio (OR) = 2.53, 95% confidence interval (CI) = 1.06-6.03], smoking during pregnancy (OR = 4.64, 95% CI = 1.97-10.95) and chronic diseases (OR for hypertension = 3.70, 95% CI = 2.25-6.06, OR for others = 2.04, 95% CI = 1.09 3.83). CONCLUSION: The findings indicate that in addition to maternal age, history of LBW deliveries; smoking during pregnancy and chronic diseases are significant determinants of LBW in this population. This is consistent with national and international findings indicating that maternal variables and risk behaviors during pregnancy play important roles on LBW. PMID- 18423040 TI - A single nucleotide polymorphism in CAPN1 associated with marbling score in Korean cattle. AB - BACKGROUND: Marbling score (MS) is the major quantitative trait that affects carcass quality in beef cattle. In this study, we examined the association between genetic polymorphisms of the micromolar calcium-activated neutral protease gene (micro-calpain, CAPN1) and carcass traits in Korean cattle (also known as Hanwoo). RESULTS: By direct DNA sequencing in 24 unrelated Korean cattle, we identified 39 sequence variants within exons and their flanking regions in CAPN1. Among them, 12 common polymorphic sites were selected for genotyping in the beef cattle (n = 421). Statistical analysis revealed that a polymorphism in the 3'UTR (c.2151*479C>T) showed significant association with MS (Pcor. = 0.02). CONCLUSION: Our findings suggest that polymorphisms in CAPN1 might be one of the important genetic factors involved in carcass quality in beef cattle, although it could be false positive association. PMID- 18423042 TI - Hydrodilatation, corticosteroids and adhesive capsulitis: a randomized controlled trial. AB - BACKGROUND: Hydrodilatation of the glenohumeral joint is by several authors reported to improve shoulder pain and range of motion for patients with adhesive capsulitis. Procedures described often involve the injection of corticosteroids, to which the reported treatment effects may be attributed. Any important contribution arising from the hydrodilatation procedure itself remains to be demonstrated. METHODS: In this randomized trial, a hydrodilatation procedure including corticosteroids was compared with the injection of corticosteroids without dilatation. Patients were given three injections with two-week intervals, and all injections were given under fluoroscopic guidance. Outcome measures were the Shoulder Pain and Disability Index (SPADI) and measures of active and passive range of motion. Seventy-six patients were included and groups were compared six weeks after treatment. The study was designed as an open trial. RESULTS: The groups showed a rather similar degree of improvement from baseline. According to a multiple regression analysis, the effect of dilatation was a mean improvement of 3 points (confidence interval: -5 to 11) on the SPADI 0-100 scale. T-tests did not demonstrate any significant between-group differences in range of motion. CONCLUSION: This study did not identify any important treatment effects resulting from three hydrodilatations that included steroid compared with three steroid injections alone. TRIAL REGISTRATION: The study is registered in Current Controlled Trials with the registration number ISRCTN90567697. PMID- 18423044 TI - Skin mucormycosis presenting as an erythema-nodosum-like rash in a renal transplant recipient: a case report. AB - INTRODUCTION: Cutaneous mucormycosis is a rare entity related to kidney transplantation. It usually presents with ecthyma-like lesions and black necrotic cellulitis. We report an unusual case of primary cutaneous mucormycosis presenting as erythema-nodosum-like lesions in a woman who had received a renal transplant. CASE PRESENTATION: A 49-year-old woman with diabetes received a living-unrelated kidney transplant. Her clinical course was uneventful for the first six months after transplantation. She then developed multiple, painful, erythema-nodosum-like lesions on her right leg and thigh following an episode of minor trauma. Mucormycosis was diagnosed by skin biopsy. Microscopic examination also showed panniculitis. The patient was treated successfully with amphotericin B and surgical resection. To our knowledge, this is the first description of primary cutaneous mucormycosis with erythema-nodosum-like lesions and panniculitis after renal transplantation. CONCLUSION: Cutaneous mucormycosis should be considered in the differential diagnosis when a kidney transplant recipient develops erythema-nodosum-like lesions with panniculitis. PMID- 18423043 TI - Diversification and adaptive sequence evolution of Caenorhabditis lysozymes (Nematoda: Rhabditidae). AB - BACKGROUND: Lysozymes are important model enzymes in biomedical research with a ubiquitous taxonomic distribution ranging from phages up to plants and animals. Their main function appears to be defence against pathogens, although some of them have also been implicated in digestion. Whereas most organisms have only few lysozyme genes, nematodes of the genus Caenorhabditis possess a surprisingly large repertoire of up to 15 genes. RESULTS: We used phylogenetic inference and sequence analysis tools to assess the evolution of lysozymes from three congeneric nematode species, Caenorhabditis elegans, C. briggsae, and C. remanei. Their lysozymes fall into three distinct clades, one belonging to the invertebrate-type and the other two to the protist-type lysozymes. Their diversification is characterised by (i) ancestral gene duplications preceding species separation followed by maintenance of genes, (ii) ancestral duplications followed by gene loss in some of the species, and (iii) recent duplications after divergence of species. Both ancestral and recent gene duplications are associated in several cases with signatures of adaptive sequence evolution, indicating that diversifying selection contributed to lysozyme differentiation. Current data strongly suggests that genetic diversity translates into functional diversity. CONCLUSION: Gene duplications are a major source of evolutionary innovation. Our analysis provides an evolutionary framework for understanding the diversification of lysozymes through gene duplication and subsequent differentiation. This information is expected to be of major value in future analysis of lysozyme function and in studies of the dynamics of evolution by gene duplication. PMID- 18423046 TI - Benchmarking of hospital information systems: monitoring of discharge letters and scheduling can reveal heterogeneities and time trends. AB - BACKGROUND: Monitoring of hospital information system (HIS) usage can provide insights into best practices within a hospital and help to assess time trends. In terms of effort and cost of benchmarking, figures derived automatically from the routine HIS system are preferable to manual methods like surveys, in particular for repeated analysis. METHODS: Due to relevance for quality management and efficient resource utilization we focused on time-to-completion of discharge letters (assessed by CT-plots) and usage of patient scheduling. We analyzed these parameters monthly during one year at a major university hospital in Germany. RESULTS: We found several distinct patterns of discharge letter documentation indicating a large heterogeneity of HIS usage between different specialties (completeness 51 - 99%, delays 0 - 90 days). Overall usage of scheduling increased during the observation period by 62%, but again showed a considerable variation between departments. CONCLUSION: Regular monitoring of HIS key figures can contribute to a continuous HIS improvement process. PMID- 18423045 TI - The usefulness of twenty-four molecular markers in predicting treatment outcome with combination therapy of amodiaquine plus sulphadoxine-pyrimethamine against falciparum malaria in Papua New Guinea. AB - BACKGROUND: In Papua New Guinea (PNG), combination therapy with amodiaquine (AQ) or chloroquine (CQ) plus sulphadoxine-pyrimethamine (SP) was introduced as first line treatment against uncomplicated malaria in 2000. METHODS: We assessed in vivo treatment failure rates with AQ+SP in two different areas in PNG and twenty four molecular drug resistance markers of Plasmodium falciparum were characterized in pre-treatment samples. The aim of the study was to investigate the association between infecting genotype and treatment response in order to identify useful predictors of treatment failure with AQ+SP. RESULTS: In 2004, Day 28 treatment failure rates for AQ+SP were 29% in the Karimui and 19% in the South Wosera area, respectively. The strongest independent predictors for treatment failure with AQ+SP were pfmdr1 N86Y (OR = 7.87, p < 0.01) and pfdhps A437G (OR = 3.44, p < 0.01). Mutations found in CQ/AQ related markers pfcrt K76T, A220S, N326D, and I356L did not help to increase the predictive value, the most likely reason being that these mutations reached almost fixed levels. Though mutations in SP related markers pfdhfr S108N and C59R were not associated with treatment failure, they increased the predictive value of pfdhps A437G. The difference in treatment failure rate in the two sites was reflected in the corresponding genetic profile of the parasite populations, with significant differences seen in the allele frequencies of mutant pfmdr1 N86Y, pfmdr1 Y184F, pfcrt A220S, and pfdhps A437G. CONCLUSION: The study provides evidence for high levels of resistance to the combination regimen of AQ+SP in PNG and indicates which of the many molecular markers analysed are useful for the monitoring of parasite resistance to combinations with AQ+SP. PMID- 18423047 TI - High-dose-rate brachytherapy for soft tissue sarcoma in children: a single institution experience. AB - PURPOSE: To report our experience treating soft tissue sarcoma (STS) with high dose rate brachytherapy alone (HBRT) or in combination with external beam radiotherapy (EBRT) in pediatric patients. METHODS AND MATERIALS: Eighteen patients, median age 11 years (range 2 - 16 years) with grade 2-3 STS were treated with HBRT using Ir-192 in a interstitial (n = 14) or intracavitary implant (n = 4). Eight patients were treated with HBRT alone; the remaining 10 were treated with a combination of HBRT and EBRT. RESULTS: After a median follow up of 79.5 months (range 12 - 159), 14 patients were alive and without evidence of disease (5-year overall survival rate 84.5%). There were no local or regional failures in the group treated with HBRT alone. One patient developed distant metastases at 14 months and expired after 17 months. In the combined HBRT and EBRT group, there was 1 local failure (22 months), and 3 patients developed pulmonary metastatic disease 18, 38 and 48 months after diagnosis and no these patients were alive at the time of this report. The overall local control to HBRT alone and HBRT plus EBRT were 100 and 90%, respectively. The acute affects most common were local erythema and wound dehiscence in 6 (33%) and 4 (22%) patients. Late effects were observed in 3 patients (16.5%). CONCLUSION: Excellent local control with tolerable side effects have been observed in a small group of paediatric patients with STS treated by HBRT alone or in combination with EBRT. PMID- 18423048 TI - Expression profiling with RNA from formalin-fixed, paraffin-embedded material. AB - BACKGROUND: Molecular characterization of breast and other cancers by gene expression profiling has corroborated existing classifications and revealed novel subtypes. Most profiling studies are based on fresh frozen (FF) tumor material which is available only for a limited number of samples while thousands of tumor samples exist as formalin-fixed, paraffin-embedded (FFPE) blocks. Unfortunately, RNA derived of FFPE material is fragmented and chemically modified impairing expression measurements by standard procedures. Robust protocols for isolation of RNA from FFPE material suitable for stable and reproducible measurement of gene expression (e.g. by quantitative reverse transcriptase PCR, QPCR) remain a major challenge. RESULTS: We present a simple procedure for RNA isolation from FFPE material of diagnostic samples. The RNA is suitable for expression measurement by QPCR when used in combination with an optimized cDNA synthesis protocol and TaqMan assays specific for short amplicons. The FFPE derived RNA was compared to intact RNA isolated from the same tumors. Preliminary scores were computed from genes related to the ER response, HER2 signaling and proliferation. Correlation coefficients between intact and partially fragmented RNA from FFPE material were 0.83 to 0.97. CONCLUSION: We developed a simple and robust method for isolating RNA from FFPE material. The RNA can be used for gene expression profiling. Expression measurements from several genes can be combined to robust scores representing the hormonal or the proliferation status of the tumor. PMID- 18423049 TI - Age differences in mental health literacy. AB - BACKGROUND: The community's knowledge and beliefs about mental health problems, their risk factors, treatments and sources of help may vary as a function of age. METHODS: Data were taken from an epidemiological survey conducted during 2003 2004 with a national clustered sample of Australian adults aged 18 years and over. Following the presentation of a vignette describing depression (n = 1001) or schizophrenia (n = 997), respondents were asked a series of questions relating to their knowledge and recognition of the disorder, beliefs about the helpfulness of treating professionals and medical, psychological and lifestyle treatments, and likely causes. RESULTS: Participant age was coded into five categories and cross-tabulated with mental health literacy variables. Comparisons between age groups revealed that although older adults (70+ years) were poorer than younger age groups at correctly recognising depression and schizophrenia, young adults (18-24 years) were more likely to misidentify schizophrenia as depression. Differences were also observed between younger and older age groups in terms of beliefs about the helpfulness of certain treating professionals and medical and lifestyle treatments for depression and schizophrenia, and older respondents were more likely to believe that schizophrenia could be caused by character weakness. CONCLUSION: Differences in mental health literacy across the adult lifespan suggest that more specific, age appropriate messages about mental health are required for younger and older age groups. The tendency for young adults to 'over identify' depression signals the need for awareness campaigns to focus on differentiation between mental disorders. PMID- 18423051 TI - Environmental influences on familial discordance of phenotype in people with homocystinuria: a case report. AB - INTRODUCTION: Non-heritable factors may have an influence on the clinical expression of monogenic inherited metabolic diseases. CASE PRESENTATION: This is a case report of a man whose mother had been diagnosed late in childhood with pyridoxine responsive homocystinuria with lens dislocation and neurodevelopmental delay. These severe complications were not observed in her son who was pyridoxine unresponsive but who had been treated appropriately since early infancy. CONCLUSION: The phenotype of people with homocystinuria can be discordant within a family, with variability in metabolic and clinical expression depending upon both the genotype and therapeutic interventions. Offspring of people with homocystinuria should be screened in early infancy and, if positive, treated appropriately whether they have pyridoxine responsive or unresponsive disease. PMID- 18423050 TI - An interactive workshop plus locally adapted guidelines can improve general practitioners asthma management and knowledge: a cluster randomised trial in the Australian setting. AB - BACKGROUND: A cluster randomised trial was conducted to determine the effectiveness of locally adapted practice guidelines and education about paediatric asthma management, delivered to general practitioners (GPs) in small group interactive workshops. METHODS: Twenty-nine practices were randomly allocated to one of three study arms. Australian asthma management guidelines were adapted to accommodate characteristics of the local area. GPs in the intervention arm (Group 1, n = 18 GPs) participated in a small group based education program and were provided with the adapted guidelines. One control arm (Group 2, n = 18 GPs) received only the adapted guidelines, while the other control arm (Group 3, n = 15 GPs) received an unrelated education intervention. GPs' knowledge, attitudes and management of paediatric asthma was assessed. RESULTS: Post intervention, intervention arm GPs were no more likely to provide a written asthma action plan, but were better able to assess the severity of asthma attack (Group 1vs Group 2 p = 0.05 and Group 1 vs Group 3 p = 0.01), better able to identify patients at high risk of severe attack (Group 1vs Group 3 p = 0.06), and tended to score higher on the asthma knowledge questionnaire (Group 1 vs Group 2 p = 0.06 and Group 1 vs Group 3 p = 0.2). Most intervention arm GPs felt more confident than control GPs to manage acute asthma attack and ongoing management of infrequent episodic asthma. CONCLUSION: Using interactive small group workshops to disseminate locally adapted guidelines was associated with improvement in GP's knowledge and confidence to manage asthma, but did not change GP's self-reported provision of written action plans. PMID- 18423052 TI - Bilateral giant femoropopliteal artery aneurysms: a case report. AB - INTRODUCTION: Popliteal artery aneurysms are the most common peripheral arterial aneurysms, and are frequently bilateral. Acute limb ischemia, rupture and compression phenomena can complicate these aneurysms when the diameter exceeds 2 cm. CASE PRESENTATION: We report an 82-year-old male patient with two giant femoropopliteal aneurysms, 10.5 and 8.5 cm diameters, managed in our institution. Both aneurysms were resected and a polytetrafluoroethylene (PTFE) femoropopliteal interposition graft was placed successfully. Management and literature review are discussed. CONCLUSION: We believe this is the first report in the medical literature of bilateral giant femoropopliteal aneurysms. PMID- 18423053 TI - DNA demethylation-dependent enhancement of toll-like receptor-2 gene expression in cystic fibrosis epithelial cells involves SP1-activated transcription. AB - BACKGROUND: The clinical course of cystic fibrosis (CF) is characterized by recurrent pulmonary infections and chronic inflammation. We have recently shown that decreased methylation of the toll-like receptor-2 (TLR2) promoter leads to an apparent CF-related up-regulation of TLR2. This up-regulation could be responsible, in part, for the CF-associated enhanced proinflammatory responses to various bacterial products in epithelial cells. However, the molecular mechanisms underlying DNA hypomethylation-dependent enhancement of TLR2 expression in CF cells remain unknown. RESULTS: The present study indicates that there is a specific CpG region (CpG#18-20), adjacent to the SP1 binding site that is significantly hypomethylated in several CF epithelial cell lines. These CpGs encompass a minimal promoter region required for basal TLR2 expression, and suggests that CpG#18-20 methylation regulates TLR2 expression in epithelial cells. Furthermore, reporter gene analysis indicated that the SP1 binding site is involved in the methylation-dependent regulation of the TLR2 promoter. Inhibition of SP1 with mithramycin A decreased TLR2 expression in both CF and 5-azacytidine treated non-CF epithelial cells. Moreover, even though SP1 binding was not affected by CpG methylation, SP1-dependent transcription was abolished by CpG methylation. CONCLUSION: This report implicates SP1 as a critical component of DNA demethylation-dependent up-regulation of TLR2 expression in CF epithelial cells. PMID- 18423056 TI - Maize genome in motion. AB - A report on the Maize Genetics Conference held in Washington DC, USA, 27 February 1 March, 2008. PMID- 18423055 TI - Gene polymorphisms of superoxide dismutases and catalase in diabetes mellitus. AB - BACKGROUND: Reactive oxygen species generated by hyperglycaemia modify structure and function of lipids, proteins and other molecules taking part in chronic vascular changes in diabetes mellitus (DM). Low activity of scavenger enzymes has been observed in patients with DM. Protective role of scavenger enzymes may be deteriorated by oxidative stress. This study was undertaken to investigate the association between gene polymorphisms of selected antioxidant enzymes and vascular complications of DM. RESULTS: Significant differences in allele and genotype distribution among T1DM, T2DM and control persons were found in SOD1 and SOD2 genes but not in CAT gene (p < 0,01). Serum SOD activity was significantly decreased in T1DM and T2DM subjects compared to the control subjects (p < 0,05). SOD1 and SOD2 polymorphisms may affect SOD activity. Serum SOD activity was higher in CC than in TT genotype of SOD2 gene (p < 0,05) and higher in AA than in CC genotype of SOD1 gene (p < 0,05). Better diabetes control was found in patients with CC than with TT genotype of SOD2 gene. Significantly different allele and genotype frequencies of SOD2 gene polymorphism were found among diabetic patients with macroangiopathy and those without it. No difference was associated with microangiopathy in all studied genes. CONCLUSION: The results of our study demonstrate that oxidative stress in DM can be accelerated not only due to increased production of ROS caused by hyperglycaemia but also by reduced ability of antioxidant defense system caused at least partly by SNPs of some scavenger enzymes. PMID- 18423054 TI - Genome-wide subcellular localization of putative outer membrane and extracellular proteins in Leptospira interrogans serovar Lai genome using bioinformatics approaches. AB - BACKGROUND: In bacterial pathogens, both cell surface-exposed outer membrane proteins and proteins secreted into the extracellular environment play crucial roles in host-pathogen interaction and pathogenesis. Considerable efforts have been made to identify outer membrane (OM) and extracellular (EX) proteins produced by Leptospira interrogans, which may be used as novel targets for the development of infection markers and leptospirosis vaccines. RESULT: In this study we used a novel computational framework based on combined prediction methods with deduction concept to identify putative OM and EX proteins encoded by the Leptospira interrogans genome. The framework consists of the following steps: (1) identifying proteins homologous to known proteins in subcellular localization databases derived from the "consensus vote" of computational predictions, (2) incorporating homology based search and structural information to enhance gene annotation and functional identification to infer the specific structural characters and localizations, and (3) developing a specific classifier for cytoplasmic proteins (CP) and cytoplasmic membrane proteins (CM) using Linear discriminant analysis (LDA). We have identified 114 putative EX and 63 putative OM proteins, of which 41% are conserved or hypothetical proteins containing sequence and/or protein folding structures similar to those of known EX and OM proteins. CONCLUSION: Overall results derived from the combined computational analysis correlate with the available experimental evidence. This is the most extensive in silico protein subcellular localization identification to date for Leptospira interrogans serovar Lai genome that may be useful in protein annotation, discovery of novel genes and understanding the biology of Leptospira. PMID- 18423057 TI - Saccharomyces boulardii fungaemia in an intensive care unit patient treated with caspofungin. AB - We describe a case of Saccharomyces boulardii fugaemia in a critically ill patient with septic shock treated with a probiotic agent containing this yeast. We attributed this fugaemia to gut translocation. Our use of caspofugin yielded excellent results. PMID- 18423058 TI - The pursuit of a high central venous oxygen saturation in sepsis: growing concerns. AB - In this issue of Critical Care, Dutch investigators report that, in a cohort of patients with sepsis/septic shock admitted to three different intensive care units (ICUs), low central venous oxygen saturation (ScvO2) was uncommon at the time of ICU admission, and hospital mortality was <30%. Their findings, taken together with those of recent reports from Australia and New Zealand (ANZ), raise serious concerns about the utility of early goal directed therapy (EGDT) outside the context of the original trial. Despite inclusion of EGDT into the Surviving Sepsis Guidelines, in response to growing uncertainty, ANZ and US investigators will soon begin randomization of patients into two large multicentre trials comparing EGDT to standard therapy. Until such studies are completed, basing international treatment guidelines on a single centre study performed in what may turn out to be a highly atypical environment would seem premature. PMID- 18423059 TI - Delirium assessment in the intensive care unit: patient population matters. AB - The high prevalence and pervasive impact of delirium in critically ill patients has been demonstrated in multiple studies. Subsequently there has grown a body of literature regarding delirium assessment in critical illness. The present commentary briefly discusses delirium screening in an intensive care unit environment. PMID- 18423060 TI - Plant immunity from A to Z. AB - A report of The Keystone Symposium on Plant Innate Immunity, Keystone, USA, 10-15 February 2008. PMID- 18423062 TI - Insulin, intracerebral glucose and bedside biochemical monitoring utilizing microdialysis. AB - Following subarachnoid hemorrhage, hyperglycemia is strongly associated with complications and with impaired neurological recovery. Targeted insulin therapy for glycemic control might, on the contrary, have harmful effects by causing too low cerebral glucose levels. The study published by Schlenk and colleagues in the previous issue of Critical Care shows that insulin caused a significant decrease in the interstitial cerebral glucose concentration although the blood glucose level remained unaffected. Since several studies utilizing various analytical techniques have shown that cerebral blood flow and cerebral glucose uptake and metabolism are insulin-independent processes, the observation remains unexplained. PMID- 18423063 TI - Prehospital therapeutic hypothermia in cardiac arrest: will there ever be evidence? PMID- 18423061 TI - Clinical review: airway hygiene in the intensive care unit. AB - Maintenance of airway secretion clearance, or airway hygiene, is important for the preservation of airway patency and the prevention of respiratory tract infection. Impaired airway clearance often prompts admission to the intensive care unit (ICU) and can be a cause and/or contributor to acute respiratory failure. Physical methods to augment airway clearance are often used in the ICU but few are substantiated by clinical data. This review focuses on the impact of oral hygiene, tracheal suctioning, bronchoscopy, mucus-controlling agents, and kinetic therapy on the incidence of hospital-acquired respiratory infections, length of stay in the hospital and the ICU, and mortality in critically ill patients. Available data are distilled into recommendations for the maintenance of airway hygiene in ICU patients. PMID- 18423064 TI - Patient-level glucose reporting: averages, episodes, or something in between? AB - The article by Van Herpe and colleagues in the previous issue of Critical Care describes the glycemic penalty index (GPI), which weights both hyperglycemic and hypoglycemic blood glucose measurements commensurate to their clinically significant difference from target. Although certain adverse consequences result from isolated severe hyperglycemic episodes, several specific outcomes depend upon overall hyperglycemia. In contrast, although mortality has been related epidemiologically to overall low blood glucose, specific negative outcomes may depend upon isolated episodes. Capturing both hypoglycemia and hyperglycemia in a single index will be shown to be useful if the GPI enables us to better define insulin strategies, outcomes, and targets. PMID- 18423065 TI - Vasopressin in vasodilatory shock: is the heart in danger? AB - In patients with hyperdynamic hemodynamics, infusing arginine vasopressin (AVP) in advanced vasodilatory shock is usually accompanied by a decrease in cardiac output and in visceral organ blood flow. Depending on the infusion rate, this vasoconstriction also reduces coronary blood flow despite an increased coronary perfusion pressure. In a porcine model of transitory myocardial ischemia-induced left ventricular dysfunction, Muller and colleagues now report that the AVP related coronary vaso-constriction may impede diastolic relaxation while systolic contraction remains unaffected. Although any AVP-induced myocardial ischemia undoubtedly is a crucial safety issue, these findings need to be discussed in the context of the model design, the dosing of AVP as well as the complex direct, afterload-independent and systemic, vasoconstriction-related effects on the heart. PMID- 18423066 TI - Chromatin organization and expression. AB - A report on the 29th Lorne Genome Conference on the Organization and Expression of the Genome, Lorne, Australia, 17-21 February 2008. PMID- 18423067 TI - Competing risks models and time-dependent covariates. AB - New statistical models for analysing survival data in an intensive care unit context have recently been developed. Two models that offer significant advantages over standard survival analyses are competing risks models and multistate models. Wolkewitz and colleagues used a competing risks model to examine survival times for nosocomial pneumonia and mortality. Their model was able to incorporate time-dependent covariates and so examine how risk factors that changed with time affected the chances of infection or death. We briefly explain how an alternative modelling technique (using logistic regression) can more fully exploit time-dependent covariates for this type of data. PMID- 18423068 TI - KL-6 in acute lung injury: will it leave its mark? AB - Studies have indicated that measuring biochemical measures of epithelial injury in plasma and alveolar fluid may be useful in predicting outcome in acute lung injury. The present commentary briefly reviews the evidence supporting the use of these biochemical biomarkers of epithelial injury in acute lung injury, and in particular KL-6, as well as their limitations. The article additionally proposes the need for physiological markers of epithelial function to complement current biochemical biomarkers. PMID- 18423069 TI - Lost in translation? The pursuit of lung-protective ventilation. AB - Acute lung injury (ALI) and the acute respiratory distress syndrome (ARDS) remain important causes of morbidity and mortality in the critically ill patient, with far-reaching short-term and long-term implications for individual patients and for healthcare providers. It is well accepted that mechanical ventilation can worsen lung injury, potentially worsening systemic organ function, and can thus impact on mortality in acute lung injury (ALI)/ARDS. Unfortunately, although the concept of minimizing such damage via lung-protective ventilatory strategies is widely acknowledged, effective integration of such an approach into clinical practice remains more elusive. The study by the Irish Critical Care Trials Group published in the previous edition of Critical Care describes a 10-week real-life survey of all intensive care unit admissions across Ireland, detailing for the first time the epidemiology of ALI/ARDS in this population and clinician's attempts to deliver lung-protective ventilation. The authors also report hypothesis-generating data on the implications of statin use in this population. The present commentary reviews aspects of this work, with particular attention to the implementation of low-tidal-volume/lung-protective ventilatory strategies in ALI/ARDS. PMID- 18423070 TI - Vasopressor stays vasopressor and inotrope stays inotrope! PMID- 18423071 TI - Cancer stem cell heterogeneity in hereditary breast cancer. AB - The cancer stem cell hypothesis proposes that tumors arise in stem or progenitor cells generating in tumors driven by a subcomponent that retains cancer stem cell properties. Recent evidence supports the hypothesis that the BRCA1 gene involved in hereditary breast cancer plays a role in breast stem cell function. Furthermore, studies using mouse BRCA1 knockout models provide evidence for the existence of heterogeneous cancer stem cell populations in tumors generated in these mice. Although these populations may arise from different stem/progenitor cells, they share the expression of a common set of stem cell regulatory genes and show similar characteristics in in vitro mammosphere assays and xenograft models. Furthermore, these 'cancer stem cells' display resistance to chemotherapeutic agents. These studies suggest that breast tumors may display intertumor stem cell heterogeneity. Despite this heterogeneity, cancer stem cells may share common characteristics that can be used for their identification and for therapeutic targeting. PMID- 18423072 TI - Sickness behavior, its mechanisms and significance. AB - Recent studies have begun to clarify the pathogenesis of sickness behavior. Cytokines released by macrophages, dendritic cells and mast cells act on the brain to trigger behavioral changes in infected animals. The major cytokines, interleukin-1, tumor necrosis factor alpha, and others, all act on the hypothalamus to provoke alterations in the normal homeostatic condition. These include elevated body temperature, increased sleep, and loss of appetite as well as major alterations in lipid and protein metabolism leading to significant weight loss. Some of these changes are clearly directed towards enhancing the normal immune responses. The benefits of others such as appetite loss are unclear. It is also important to recognize that other animals may recognize sickness behavior as a sign of weakness and mark the victim out for targeting by predators. As a result, some prey species may work very hard to mask their sickness, a response that serves to complicate veterinary diagnosis. PMID- 18423073 TI - Generation of hydrogen peroxide by a low molecular weight compound in whey of Holstein dairy cows. AB - Using an ultrafiltration membrane (molecular cut-off, 3,000), low molecular weight compounds in bovine milk were collected (YM-3 filtrate). A hydrogen peroxide (H2O2)-like substance was generated in the YM-3 filtrate. This substance was undetected at 0 h, but increased in a time-dependent manner, peaking after 2 h of incubation at 38 degrees C. After incubating the YM-3 filtrate with catalase and lactoperoxidase, the signal showing the presence of this substance disappeared. The substance was quantified using one chemiluminescence and three colorimetric H2O2 detection systems. In all systems, their estimates were within the same range. The amount of substance, as estimated by the chemiluminescence H2O2 detection system, was correlated with that estimated by the other three colorimetric systems (r=0.98, 0.95 and 0.87). The substance was eluted at the same position as H2O2 by gel filtration on Superdex 30. Thus, the substance had the same characteristics as H2O2. An H2O2-generating substance in either the YM-3 filtrate or whey had a molecular mass of about 600. In this study, we clarify that bovine milk is capable of generating H2O2 by utilizing a low molecular weight compound. Thus, we present a new type of H2O2-supplying system in bovine milk. PMID- 18423074 TI - The prevalence and workplace costs of adult attention deficit hyperactivity disorder in a large manufacturing firm. AB - BACKGROUND: Little is known about the effects of adult attention deficit hyperactivity disorder (ADHD) on work performance or accidents-injuries.MethodA survey was administered in 2005 and 2006 to employees of a large manufacturing firm to assess the prevalence and correlates of adult ADHD. Respondents (4,140 in 2005, 4,423 in 2006, including 2,656 in both surveys) represented 35-38% of the workforce. ADHD was assessed with the World Health Organization (WHO) Adult ADHD Self-Report Scale (ASRS), a validated screening scale for DSM-IV adult ADHD. Sickness absence, work performance and workplace accidents-injuries were assessed with the WHO Health and Work Performance Questionnaire (HPQ). RESULTS: The estimated current prevalence (standard error) of DSM-IV ADHD was 1.9% (0.4). ADHD was associated with a 4-5% reduction in work performance (chi12=9.1, p=0.001), a 2.1 relative-odds of sickness absence (chi12=6.2, p=0.013), and a 2.0 relative odds of workplace accidents-injuries (chi12=5.1, p=0.024). The human capital value (standard error) of the lost work performance associated with ADHD totaled USD 4,336 (676) per worker with ADHD in the year before interview. No data were available to monetize other workplace costs of accidents-injuries (e.g. destruction of equipment). Only a small minority of workers with ADHD were in treatment. CONCLUSIONS: Adult ADHD is a significantly impairing condition among workers. Given the low rate of treatment and high human capital costs, in conjunction with evidence from controlled trials that treatment can reduce ADHD related impairments, ADHD would seem to be a good candidate for workplace trials that evaluate treatment cost-effectiveness from the employer's perspective. PMID- 18423075 TI - The emperors of the schizophrenia polygene have no clothes. AB - A substantial body of research literature, identified by nine out of ten papers on genetics in the recent ISI research front on schizophrenia, claims to have established associations between aspects of the disease and sequence variation in specific candidate genes. These candidatures have proven unreplicated in large sibling pair linkage surveys and a targeted association study. Even if the case for an association be regarded as a lucky guess (assuming one gene in 30 000 was guessed right) the large linkage and association studies provide no evidence of sequence variation relating to psychosis at any of these gene loci. Thus this body of work must be regarded as an indicator of the extent to which the 'eye of faith' is able to discern meaning in complex data when none is present. PMID- 18423076 TI - Life history traits of Aedes caspius (Diptera: Culicidae): a laboratory study of larval stages. AB - The larval survival and development times of Aedes caspius (Diptera: Culicidae) were examined in the laboratory. These life history traits were estimated using life tables constructed for two populations, one of which had been subjected to a long-term larvicide control program. Traits were evaluated for eight different population densities. The effects of population, larval stage and larval density were investigated using a general linear model. Density was positively correlated with larval survival but did not affect development time. The fourth instar and pupae had the lowest larval survival rates. First and fourth instar larvae had the longest development times. These traits were not significantly different between the two populations. The effect of larvicide control on these traits is discussed. PMID- 18423077 TI - Thrips (Thysanoptera) identification using artificial neural networks. AB - We studied the use of a supervised artificial neural network (ANN) model for semi automated identification of 18 common European species of Thysanoptera from four genera: Aeolothrips Haliday (Aeolothripidae), Chirothrips Haliday, Dendrothrips Uzel, and Limothrips Haliday (all Thripidae). As input data, we entered 17 continuous morphometric and two qualitative two-state characters measured or determined on different parts of the thrips body (head, pronotum, forewing and ovipositor) and the sex. Our experimental data set included 498 thrips specimens. A relatively simple ANN architecture (multilayer perceptrons with a single hidden layer) enabled a 97% correct simultaneous identification of both males and females of all the 18 species in an independent test. This high reliability of classification is promising for a wider application of ANN in the practice of Thysanoptera identification. PMID- 18423078 TI - Effects of forest-dune ecotone management on the endangered heath grasshopper, Chorthippus vagans (Orthoptera: Acrididae). AB - Dry, oligotrophic ecosystems are highly threatened in Europe due to massive changes in land use and eutrophication. The conservation of these xeric habitats has received much attention, whereas the ecotones between xeric habitats and other habitat types are often disregarded. One species which mainly inhabits the transition zone between pine forests and adjacent xeric habitats is the heath grasshopper, Chorthippus vagans. This species is endangered in large parts of Europe. One of the largest populations in northern Germany is found on a degraded inland dune near Hanover. This population is threatened by dense growth of deciduous trees and litter accumulation. We analyzed changes in the distribution of this population after the implementation of conservation measures (thinning out the forest and removal of leaf litter). Moreover, we examined dispersal distances of the species in order to assess its colonization potential. We also studied the microhabitat preferences of C. vagans to assess key factors influencing its local distribution. Our data show a substantial growth in population size, which might be a consequence of the conservation measures. New patches on the dune were colonized, promoting dispersal between the subpopulations. We propose that restoration of forest-dune ecotones should be considered more often in landscape planning and conservation management. PMID- 18423079 TI - Screening for hearing loss and middle-ear effusion in school-age children, using transient evoked otoacoustic emissions: a feasibility study. AB - INTRODUCTION: The characteristics of otoacoustic emissions that make them ideally suited for universal newborn hearing loss screening could potentially be useful for the screening of older children. This study was performed in order to assess the role of otoacoustic emissions in a screening programme for middle-ear disorders and hearing loss in school-age children. METHODS: Cross-sectional, preliminary screening study. SETTING: Primary schools of Argolida municipality, south-east Greece, between December 2004 and March 2005. PATIENT SELECTION AND RECRUITMENT: All the primary school students of Argolida were invited, by press releases and individually by their teachers, to attend a session of otological and audiological screening. RESULTS: One hundred and ninety-six children were evaluated using transient evoked otoacoustic emissions. Twenty per cent failed in both ears, while in 32 per cent otoacoustic emissions could not be produced in at least one ear. Younger children had higher rates of absent transient evoked otoacoustic emissions. The absence of otoacoustic emissions was highly correlated with tympanic membrane changes seen on otoscopy and the presence of a type B tympanogram. As a single screening modality, otoacoustic emissions had a 100 per cent sensitivity in diagnosing hearing loss worse than 30 dB, and a 90 per cent sensitivity and 64 per cent specificity in diagnosing hearing loss worse than 25 dB, which did not improve by adding tympanometry to the screening protocol. CONCLUSION: These results strongly suggest the potential usefulness of otoacoustic emission testing in screening school-age children for hearing loss. Further studies, taking into account cost-effectiveness issues, are indicated. PMID- 18423080 TI - Pre-operative prediction of 'dry taps'. AB - OBJECTIVES: The treatment of children with 'glue ear' often presents surgeons with the question of whether or not to insert a grommet when myringotomy reveals no fluid in the middle ear. We present a study designed to assess which factors contribute to the presence of a 'dry tap'. DESIGN: We prospectively gathered data from a cohort of 280 children (504 myringotomies). The cohort included two subgroups, one received halothane and nitrous oxide anaesthesia, and the other received enflurane anaesthesia. SETTING: The ENT department of a district general hospital. PARTICIPANTS: Children (aged less than 17 years) requiring myringotomy. MAIN OUTCOME MEASURES: The presence of a 'glue' or dry tap at myringotomy was documented. We also recorded data on the following: pre- and post-induction tympanometry; age; season; anaesthetic type; and the delay from listing to actual operation. RESULTS: A non type B pre-induction tympanogram and delay to operation were strong indications of finding a dry tap at surgery. CONCLUSIONS: In our study population, the proportion of dry taps at myringotomy was 18 per cent. The presence of a dry tap was rarely due to the induction of anaesthesia. Multivariate analysis revealed that the combination of factors most likely to predict a dry tap were non type B tympanogram and delay to operation. PMID- 18423081 TI - Quinsy trainer. PMID- 18423082 TI - Magnetic resonance identification of an accessory submandibular duct and gland: an unusual variant. PMID- 18423083 TI - Use of an optical surface scanner in assessment of outcome following rhinoplasty surgery. AB - OBJECTIVE: To demonstrate the use of an optical surface scanner, with associated software, in the assessment of rhinoplasty patients, and to discuss the possible clinical applications of this technology in the future. DESIGN: Case study analysis of pre- and post-operative scans of a patient undergoing septorhinoplasty at Whipps Cross University Hospital, London, UK. SUBJECT: A 21 year-old man undergoing septorhinoplasty underwent pre-operative optical surface scanning of his face. The scans were repeated at one week and one year post operatively. Software developed at University College London was then used to analyse the scans. RESULTS: The scans clearly showed that the man's dorsal hump had been well reduced and the nose straightened, with a resulting 1600 mm3 gain on the right side and a 1000 mm3 loss on the left side of the nose. Tip projection had also been achieved. CONCLUSION: This technique allowed objective quantification of facial features and analysis of change. It may well prove useful in the future in predicting change following surgical intervention. PMID- 18423084 TI - Squamous cell carcinoma of the nasal vestibule: a 20-year case series and literature review. AB - Squamous cell carcinoma of the nasal vestibule is a rare disease with significant morbidity and mortality, and a five-year recurrence-free survival rate of between 42 and 92 per cent. There are several staging systems: the American Joint Committee on Cancer (AJCC) skin and nasoethmoid complex system, the Union International Centre Cancer (UICC) nasal fossa system, and Wang's system. Treatment options include radiotherapy or surgery for early lesions, but more advanced cases require radical surgery with post-operative radiotherapy. We present a case series spanning the last 20 years in one centre, and we compare this series with cases reported in the literature, paying particular attention to staging, treatment and outcome. We found that patients with tumours staged T2 or T3 (Wang system) who received radiotherapy alone did poorly in comparison with those who received surgery or surgery and radiotherapy. PMID- 18423085 TI - Causes of childhood deafness in Pukhtoonkhwa Province of Pakistan and the role of consanguinity. AB - BACKGROUND: Deafness is the hidden disability of childhood, and leads to poor educational and employment prospects. There is little published information on deafness in Pakistan. Profound hearing impairment is more prevalent in countries where consanguineous marriages are common, such as Pakistan. This study aimed to assess causes of childhood deafness and association with parental consanguinity, within deaf and hearing children in the Peshawar district of Pukhtoonkhwa Province, Pakistan. METHODS: One hundred and forty deaf children were identified from two schools for deaf children within the Peshawar district. These children were assessed via audiology, otoscopic examination, case note review and parental history, in order to attempt to ascertain the cause of their deafness. Two hundred and twenty-one attendees at a local immunisation clinic (taken as representative of the local childhood population) were also screened for hearing impairment. Parents of both groups of children were assessed by interview and questionnaire in order to ascertain the mother and father's family relationship (i.e. whether cousins or unrelated). RESULTS: Of the 140 deaf school pupils, 92.1 per cent were profoundly hearing impaired and 7.9 per cent were severely hearing impaired. All these children had bilateral sensorineural hearing loss. A possible cause of deafness was identified in only six of these children. Parental consanguinity (i.e. first or second cousins) was established for 86.4 per cent of deaf school pupils and 59.7 per cent of immunisation clinic attendees. None of the control children were identified as having a hearing problem. CONCLUSION: The prevalence of parental consanguinity was significantly higher in deaf children compared with non-hearing impaired children. However, the study also confirmed a high rate of consanguinity within the general Peshawar community. In this setting, prevention of consanguineous unions is the only means of reducing levels of congenital hearing impairment. The current levels of hearing disability represent both a prominent public health problem and an important, potentially preventable childhood disability. PMID- 18423086 TI - Exuberant and inhibited toddlers: stability of temperament and risk for problem behavior. AB - Temperament, effortful control, and problem behaviors at 4.5 years were assessed in 72 children classified as exuberant, inhibited, and low reactive as 2-year olds. Exuberant toddlers were more positive, socially responsive to novel persons, less shy, and rated as having more problem behaviors, including externalizing and internalizing behaviors, than other children as preschoolers. Two forms of effortful control, the ability to delay a response and the ability to produce a subdominant response, were associated with fewer externalizing behaviors, whereas expressing more negative affect (relative to positive/neutral affect) when disappointed was related to more internalizing behaviors. Interaction effects implicated high levels of unregulated emotion during disappointment as a risk factor for problem behaviors in exuberant children. PMID- 18423087 TI - Salivary cortisol levels in children of low-income women with high depressive symptomatology. AB - Children (N = 324 boys, 315 girls) between the ages of 2.5 and 6 (mean age = 3.63) were identified in a house to house survey in low-income areas (income <20th percentile nationally) of urban Mexico. The Center for Epidemiologic Studies-Depression Scale was administered to mothers of all children. Salivary cortisol samples were taken in children as a measure of hypothalamic-pituitary adrenocortical (HPA) system activity at time of arrival (baseline, Time 0), 25 min after arrival (Time 1), and 50 min after arrival (Time 2). Between Time 0 and Time 1, children were administered several cognitive tests. Results of hierarchical linear modeling analyses revealed that higher levels of maternal depressive symptoms were associated with lower baseline cortisol levels in their children (p < .05), while controlling for age, gender, and time since awakening. Higher levels of maternal depressive symptoms were associated with less of an increase in salivary cortisol to the arrival of the experimenters and subsequent cognitive testing (p < .05). All results were moderated by gender, with enhanced cortisol response in girls and no response in boys. These results suggest that among very low-income families, high maternal depressive symptoms are associated with hypoactivity of the HPA system in children, particularly boys. PMID- 18423088 TI - Maternal prenatal smoking, parental antisocial behavior, and early childhood physical aggression. AB - This study investigated joint effects of maternal prenatal smoking and parental history of antisocial behavior on physical aggression between ages 17 and 42 months in a population sample of children born in Quebec (N = 1,745). An analysis of variance (ANOVA) showed significant main effects of maternal prenatal smoking and a significant interaction between maternal prenatal smoking and mother's history of antisocial behavior in the prediction of children's probability to display high and rising physical aggression. The interaction indicated that the effects of heavy smoking during pregnancy (> or =10 cigarettes/day) were greater when the mother also had a serious history of antisocial behavior. The effects remained significant after the introduction of control variables (e.g., hostile reactive parenting, family functioning, parental separation/divorce, family income, and maternal education). Another significant interaction not accounted for by control variables was observed for maternal prenatal smoking and family income, indicating more serious effects of maternal prenatal smoking under relatively low-income, conditions. Both interactions indicate critical adversities that, in combination with maternal prenatal smoking, have supra additive effects on (the development of) physical aggression during early childhood. These findings may have implications for the selection of intervention targets and strategies. PMID- 18423089 TI - Gene-environment interaction between peer victimization and child aggression. AB - Although peer victimization places children at serious risk for aggressive behavior, not all victimized children are aggressive. The diathesis-stress hypothesis of disease proposes that an environmental stressor such as peer victimization should to lead to maladjustment mostly in those individuals with preexisting genetic vulnerabilities. Accordingly, this study examined whether the link between peer victimization and child aggression is moderated by children's genetic risk for such behavior. Using a sample of 506 6-year-old twins, peer victimization was assessed through peer nominations and aggressive behavior was assessed through peer and teacher reports. Children's genetic risk for aggression was estimated as a function of their co-twin's aggression and the pair's zygosity. Genetic modeling showed that peer victimization is an environmentally driven variable that is unrelated to children's genetic disposition. Results also provided support for the notion of a gene-environment interaction between peer victimization and child's genetic risk for aggressive behavior, albeit only in girls. For boys, peer victimization was related to aggression regardless of the child's genetic risk for such behavior. Different socialization experiences in girls' compared to boys' peer groups may explain the different pattern of results for girls and boys. PMID- 18423091 TI - Longitudinal association between infant disorganized attachment and childhood posttraumatic stress symptoms. AB - The purpose of this study was to evaluate whether children with a history of disorganized attachment in infancy were more likely than children without a history of disorganized attachment to exhibit symptoms of posttraumatic stress disorder (PTSD) at school age following trauma exposure. The sample consisted of 78 8.5-year-old children from a larger, ongoing prospective study evaluating the effects of intrauterine cocaine exposure (IUCE) on children's growth and development from birth to adolescence. At the 12-month visit, children's attachment status was scored from videotapes of infant-caregiver dyads in Ainsworth's strange situation. At the 8.5-year visit, children were administered the Violence Exposure Scale-Revised, a child-report trauma exposure inventory, and the Diagnostic Interview for Children and Adolescents by an experienced clinical psychologist masked to children's attachment status and IUCE status. Sixteen of the 78 children (21%) were classified as insecure disorganized/insecure-other at 12 months. Poisson regressions covarying IUCE, gender, and continuity of maternal care indicated that disorganized attachment status at 12 months, compared with nondisorganized attachment status, significantly predicted both higher avoidance cluster PTSD symptoms and higher reexperiencing cluster PTSD symptoms. These findings suggest that the quality of early dyadic relationships may be linked to differences in children's later development of posttraumatic stress symptoms following a traumatic event. PMID- 18423090 TI - Early deprivation and home basal cortisol levels: a study of internationally adopted children. AB - Animal studies reveal that early deprivation impairs regulation of the hypothalamic-pituitary-adrenocortical (HPA) axis, potentially increasing vulnerability to stressors throughout life. To examine early deprivation effects on basal HPA axis activity in humans, basal cortisol levels were examined in 164 internationally adopted children who had experienced varying degrees of preadoption deprivation. Duration of institutional care, age at adoption, and parent ratings of preadoption neglect indexed a latent factor of Deprived Care. Adoption measures of height and weight standardized to World Health Organisation norms indexed a latent factor of Growth Delay that was viewed as another reflection of deprivation. Cortisol samples were collected 3.3-11.6 years postadoption (Md = 7.3 years) at home on 3 days approximately 30 min after wakeup and before bedtime. Both early a.m. levels and the decrease in cortisol across the day were examined. A structural equation model revealed that preadoption Deprived Care predicted Growth Delay at adoption and Growth Delay predicted higher morning cortisol levels and a larger diurnal cortisol decrease. PMID- 18423092 TI - Effortful control as a moderator of the relation between contextual risk factors and growth in adjustment problems. AB - Effortful control was examined as a moderator of the relations of three domains of contextual risk factors to growth in internalizing and externalizing problems in a community sample (N = 189) of children (8-12 years at Time 1). Socioeconomic, maternal, and environmental risk factors were examined as predictors of initial levels and growth in children's adjustment problems across 3 years. The effects of the risk factors depended on children's level of effortful control. For children lower in effortful control, socioeconomic risk was related to significantly higher initial levels of internalizing and externalizing problems and decreases over time. However, children lower in effortful control had higher levels of problems at all three time points than children higher in effortful control. Maternal risk was associated with increases in internalizing for children lower in effortful control, and environmental risk was related to increases in internalizing and externalizing problems for children lower in effortful control, but not those higher in effortful control. Children who were lower in effortful control appeared to experience more adverse effects of contextual risk than those higher in effortful control, suggesting that interventions aimed at improving children's effortful control might serve to protect children from increased risk of adjustment problems associated with contextual risk factors. PMID- 18423094 TI - Emotional difficulties in early adolescence following severe early deprivation: findings from the English and Romanian adoptees study. AB - The study assessed conduct and emotional difficulties in a group of Romanian adoptees at age 11, and serves as a follow-up to assessments made when the children were 6 years old. It was found that there was a significant increase in emotional difficulties, but not conduct problems, for the Romanian sample since age 6. It was also found that emotional difficulty was significantly more prevalent at age 11 in the Romanian group than in a within-UK adoptee group. Emotional difficulties in the Romanian adoptee group were found to be significantly and strongly related to previous deprivation-specific problems (disinhibited attachment, cognitive impairment, inattention/overactivity and quasi-autism); however, the presence of such early problems did not account fully for the onset of later emotional problems. Five contrasting hypotheses concerning possible mediators for later onset of emotional difficulties for the Romanian group were examined. No links were found to duration of deprivation or other deprivation-related indices, stresses/difficulties in the postadoption family environment, or educational attainment and self-esteem. There was some evidence that emotion recognition might play a role in the emergence of these problems, but other measures of social competence and theory of mind showed no associations with the onset of emotional problems. PMID- 18423093 TI - Face emotion labeling deficits in children with bipolar disorder and severe mood dysregulation. AB - Children with narrow phenotype bipolar disorder (NP-BD; i.e., history of at least one hypomanic or manic episode with euphoric mood) are deficient when labeling face emotions. It is unknown if this deficit is specific to particular emotions, or if it extends to children with severe mood dysregulation (SMD; i.e., chronic irritability and hyperarousal without episodes of mania). Thirty-nine NP-BD, 31 SMD, and 36 control subjects completed the emotional expression multimorph task, which presents gradations of facial emotions from 100% neutrality to 100% emotional expression (happiness, surprise, fear, sadness, anger, and disgust). Groups were compared in terms of intensity of emotion required before identification occurred and accuracy. Both NP-BD and SMD youth required significantly more morphs than controls to label correctly disgusted, surprised, fearful, and happy faces. Impaired face labeling correlated with deficient social reciprocity skills in NP-BD youth and dysfunctional family relationships in SMD youth. Compared to controls, patients with NP-BD or SMD require significantly more intense facial emotion before they are able to label the emotion correctly. These deficits are associated with psychosocial impairments. Understanding the neural circuitry associated with face-labeling deficits has the potential to clarify the pathophysiology of these disorders. PMID- 18423095 TI - Callous-unemotional traits and the emotional processing of distress cues in detained boys: testing the moderating role of aggression, exposure to community violence, and histories of abuse. AB - Callous-unemotional (CU) traits in antisocial youth have been associated with deficits in the processing of emotionally distressing stimuli in a number of past studies. In the current study, we investigated moderators of this association in a sample of 88 ethnically diverse detained boys (mean age = 15.57, SD = 1.28). Overall, emotional processing of distressing stimuli using a dot-probe task was not related to CU traits and there was no moderating effect of ethnicity. However, CU traits were related to deficits in emotional processing in youth high on aggression and youth high on exposure to community violence. Further, youth high on CU traits but with enhanced orienting to distressing stimuli had stronger histories of abuse, supporting the possibility that there may be environmentally influenced pathways in the development of these traits. PMID- 18423096 TI - Substance use and related behaviors among suburban late adolescents: the importance of perceived parent containment. AB - This study builds upon prior findings of elevated substance use among suburban high school students, examining the ramifications of different parenting dimensions on substance use and related behaviors. The sample consisted of 258 11th graders in an affluent suburban community. Parenting predictors considered included those well-studied previously such as monitoring and closeness, as well as two newer dimensions: perceived containment (stringency of anticipated reactions in reaction to negative behaviors) and perceived commitment (e.g., helping the child despite other commitments). Outcomes included self-reported substance use, delinquency, and rule breaking, as well as teacher-rated inattentiveness and school grades. Findings showed elevated substance use among these 17-year-olds compared with national norms, especially among girls. Of the parent predictors, significant unique links with multiple outcomes were found for parents' knowledge of their children's activities and perceived parental containment (stringent repercussions) in reaction to the children's substance use. Notably, students reported that their parents were much more tolerant of their substance use than of other problem behaviors such as rudeness to adults and minor acts of delinquency. Results are discussed along with the implications for practice and research. PMID- 18423097 TI - Understanding the association between adolescent marijuana use and later serious drug use: gateway effect or developmental trajectory? AB - Because marijuana use often precedes the use of other psychoactive substances, it has been characterized as a gateway to these other substances. The present study used data from both monozygotic (MZ) and dizygotic (DZ) twin pairs drawn from the National Longitudinal Study of Adolescent Youth (Add Health) to examine the "gateway effect" role of earlier marijuana on later hard drug use. Difference score analyses reveal that within-pair differences in earlier marijuana use, controlling for differences in earlier hard drug use, and peer marijuana use predicted later within-pair hard drug use differences for DZ twin pairs. In contrast, earlier differences in marijuana use among MZ twin pairs did not predict later hard drug use differences. Rather than supporting the interpretation that earlier marijuana use "triggers" later hard drug use, these results suggest that the longitudinal pattern of drug use that has been interpreted as the "gateway effect" might be better conceptualized as a genetically influenced developmental trajectory. PMID- 18423098 TI - Socioeconomic background and the developmental course of schizotypal and borderline personality disorder symptoms. AB - Low socioeconomic status (SES) background has been identified as a risk for several mental disorders. However evidence regarding SES and the developmental course of personality disorder (PD) has not been addressed. Nor is it clear whether an SES relationship to PD symptom course may be attributable to known associated risks. Further, specificity of such relationships to a particular PD diagnostic pattern independent of comorbidity with other PD or with depression has not been investigated. Data are from a general population studied longitudinally between ages 10 and 36 in four assessment waves. Effects of SES associated risks on the level of symptoms of schizotypal and borderline disorders are estimated and compared to effects on depressive symptoms. Low family SES had robust modest independent effects on both PDs over the entire age span despite substantial cumulative effects of trauma history, stressful recent life events, IQ, poor parenting, and comorbid symptoms. SES effects on depressive symptoms were generally absent, but a small "protective" effect of low SES appeared when comorbidity with PD symptoms was taken into account. Cumulatively, these risks account for developmental failures of substantial magnitude and consequence, marking the importance of understanding the remaining mechanisms of SES effects and programmatic implications for minimizing associated risk. PMID- 18423099 TI - A prospective study of child maltreatment and self-injurious behavior in a community sample. AB - In conjunction with prospective ratings of child maltreatment (i.e., sexual abuse, physical abuse, and physical neglect) and measures of dissociation and somatization, this study examined prospective pathways between child maltreatment and nonsuicidal, direct self-injurious behavior (SIB; e.g., cutting, burning, self-hitting). Ongoing participants in the Minnesota Longitudinal Study of Parents and Children (N = 164; 83 males, 81 females) completed a semistructured interview about SIB when they were 26 years old. SIB emerged as a heterogeneous and prominent phenomenon in this low-income, mixed-gender, community sample. Child sexual abuse predicted recurrent injuring (i.e., three or more events; n = 13), whereas child physical abuse appeared more salient for intermittent injuring (i.e., one to two events; n = 13). Moreover, these relations appeared largely independent of risk factors that have been associated with child maltreatment and/or SIB, including child cognitive ability, socioeconomic status, maternal life stress, familial disruption, and childhood exposure to partner violence. Dissociation and somatization were related to SIB and, to a lesser degree, child maltreatment. However, only dissociation emerged as a significant mediator of the observed relation between child sexual abuse and recurrent SIB. The findings are discussed within a developmental psychopathology framework in which SIB is viewed as a compensatory regulatory strategy in posttraumatic adaptation. PMID- 18423100 TI - Female and male antisocial trajectories: from childhood origins to adult outcomes. AB - This article reports on the childhood origins and adult outcomes of female versus male antisocial behavior trajectories in the Dunedin longitudinal study. Four antisocial behavior trajectory groups were identified among females and males using general growth mixture modeling and included life-course persistent (LCP), adolescent-onset, childhood-limited, and low trajectory groups. During childhood, both LCP females and males were characterized by social, familial and neurodevelopmental risk factors, whereas those on the adolescent-onset pathway were not. At age 32, women and men on the LCP pathway were engaging in serious violence and experiencing significant mental health, physical health, and economic problems. Females and males on the adolescent-onset pathway were also experiencing difficulties at age 32, although to a lesser extent. Although more males than females followed the LCP trajectory, findings support similarities across gender with respect to developmental trajectories of antisocial behavior and their associated childhood origins and adult consequences. Implications for theory, research, and practice are discussed. PMID- 18423101 TI - Unresolved loss in the Adult Attachment Interview: implications for marital and parenting relationships. AB - This study examined links between the unresolved loss of a significant person and current functioning in marital and parenting relationships. Participants were 80 women who had experienced loss, their husbands, and their preschool children. Unresolved loss was assessed using the Adult Attachment Interview, and individual, marital, and parenting adaptation was assessed through videotaped observations and women's self-reports. As predicted, women with unresolved loss displayed less positive emotion and more anxiety and anger with both their husbands and children, compared to women who were not unresolved. They also displayed less authoritative and more authoritarian parenting styles with their children. Yet unresolved women did not report more individual or relationship difficulties, suggesting that direct observations are needed to assess the implications of unresolved loss for family functioning. PMID- 18423102 TI - Biophysics of viral infectivity: matching genome length with capsid size. AB - In this review, we discuss recent advances in biophysical virology, presenting experimental and theoretical studies on the physical properties of viruses. We focus on the double-stranded (ds) DNA bacteriophages as model systems for all of the dsDNA viruses both prokaryotic and eukaryotic. Recent studies demonstrate that the DNA packaged into a viral capsid is highly pressurized, which provides a force for the first step of passive injection of viral DNA into a bacterial cell. Moreover, specific studies on capsid strength show a strong correlation between genome length, and capsid size and robustness. The implications of these newly appreciated physical properties of a viral particle with respect to the infection process are discussed. PMID- 18423103 TI - Impact of solifenacin on resource utilization, work productivity and health utility in overactive bladder patients switching from tolterodine ER. AB - OBJECTIVE: Assess changes in resource utilization, work and activity impairment, and health utility among OAB patients continuing to have urgency symptoms with tolterodine ER 4 mg and willing to try solifenacin 5/10 mg. RESEARCH DESIGN AND METHODS: This was an open-label, non-comparative, flexible-dosing, multicenter, 12-week study assessing the efficacy and safety of solifenacin 5/10 mg/day. Patients receiving tolterodine ER 4 mg/day for >/=4 weeks but continuing to experience residual urgency symptoms (>/=3 urgency episodes/24 h) were enrolled into the study. After a 14-day washout, patients began treatment with solifenacin 5 mg/day with dosing adjustments allowed at Weeks 4 and 8. MAIN OUTCOME MEASURES: Outcomes were assessed using the Work Productivity and Activity Impairment Questionnaire - Specific Health Problem (WPAI-SHP), Health Utilities Index (HUI), and a resource utilization questionnaire administered at Pre-Washout and Week 12. RESULTS: Patients (n=440) reported significantly fewer physician office visits (p<0.0001), UTIs (p<0.0001), and pads/diapers (p=0.0009) during the study period while receiving solifenacin 5/10 mg/day, compared with the Pre-Washout period when receiving tolterodine ER 4 mg/day. After 12 weeks of treatment with solifenacin 5/10 mg/day, patients reported a reduction in work time missed (p=0.0017), less impairment while working (p<0.0001), less overall work impairment (p<0.0001) and a reduction in activity impairment (p<0.0001) compared to Pre-Washout. There was no significant difference in health utility scores. Treatment-emergent adverse events were mostly anticholinergic in nature, and were mild to moderate in severity. CONCLUSION: Overall, solifenacin 5/10 mg/day improved work productivity, activity participation, and reduced medical care use in OAB patients who continued to have urgency symptoms with tolterodine ER 4 mg/day and wished to switch to solifenacin 5/10 mg. This was an open-label, non comparative study; therefore, further research is needed to confirm these results. PMID- 18423104 TI - Familial cold auto-inflammatory syndrome (FCAS): characterization of symptomatology and impact on patients' lives. AB - OBJECTIVE: Familial cold auto-inflammatory syndrome (FCAS), a subtype of cryopyrin-associated periodic syndromes (CAPS), is a rare, inherited disease that is virtually unknown to healthcare professionals. The aim of this patient survey was to characterize the symptomatology and evaluate the debilitating effects of FCAS on patients' daily lives. RESEARCH DESIGN AND METHODS: Patients included in a disease database consisting of 167 FCAS and Muckle-Wells syndrome (MWS) patients were provided an opportunity to voluntarily participate in a cross sectional market-based survey. Upon assessment of eligibility, individual in depth phone interviews were conducted by an independent research agency to characterize disease symptomatology, diagnosis, and disease impact on daily activities. RESULTS: Thirty patients with prior diagnosis of FCAS participated. The most common and recurring symptoms reported were rash, joint pain, chills, and fever. The majority of survey participants (90%) reported that they presented with symptoms as newborns or in early childhood - symptoms which became burdensome by school age, with patients reporting recurring symptoms and debilitating disease flares precipitated by environmental exposure to cooling temperatures. To cope with their underlying disease and to try to avoid symptomatic flares, patients reported limiting their work, school, family, and social activities. Seventy-eight percent of survey participants described an impact of the disease on their work, including absenteeism and impaired job advancement; frequently they quit their job as a consequence of their disease. Over 95% of survey participants reported that FCAS prevented participation in outdoor activities, while 83% indicated an impact on social activities, including relationships with friends and family. Limitations of this survey include the absence of a validated quality-of-life instrument, lack of correlation with patient medical records, and potential recall and responder bias. The number of participants appears small, but reflects 18% of the known FCAS population in the USA. CONCLUSIONS: FCAS causes lifelong debilitating effects that restrict patients' daily lives. Poor recognition of this rare disease among healthcare professionals leads to delayed diagnosis and inappropriate or ineffective treatment. Healthcare providers need to be made aware of this serious debilitating disease to enable accurate and timely diagnosis and more compassionate management of this lifelong condition. PMID- 18423105 TI - The role of microalbuminuria and insulin resistance as significant risk factors for white matter lesions in Japanese type 2 diabetic patients. AB - OBJECTIVE: The presence of white matter lesions (WML) is an important prognostic factor for the development of stroke. Microalbuminuria, which is associated with diabetes, has been flagged as a novel predictor for cerebrovascular events. This preliminary study was therefore designed to test the hypothesis that the presence of WML correlates with microalbuminuria and insulin resistance in patients with type 2 diabetic mellitus (DM) not receiving insulin treatment. PATIENTS AND METHODS: Based on brain magnetic resonance imaging (MRI) findings, 90 type 2 diabetic patients were divided into two groups: a WML-positive group (57+/-8 years, mean+/-SD, n=34) and a WML-negative group (57+/-6 years, n=56). The level of blood glucose was assessed by fasting plasma glucose (FPG), fasting immunoreactive insulin (F-IRI), homeostasis model assessment (HOMA) index, and hemoglobin A(1c) (HbA1c). RESULTS: The body mass index was higher in the WML positive group than in the WML-negative group (p<0.01). Plasma levels of triglycerides were higher while high-density lipoprotein cholesterol (HDL-C) was lower in the WML-positive group than in the WML-negative group (p<0.05 and p<0.0001, respectively). Fasting plasma glucose (p<0.005), insulin concentrations (p<0.0001), HOMA index (p<0.0001), and urinary albumin excretion (p<0.0001) levels were higher in the WML-positive group than in the WML-negative group. Multivariate logistic analysis revealed that WML was independently predicted by the microalbuminuria and insulin resistance (p<0.005, p<0.0005, respectively). CONCLUSION: The results of this preliminary study indicate that the presence of WML was associated with the microalbuminuria and insulin resistance in these Japanese patients with type 2 DM; larger cohort studies are warranted to confirm these findings. PMID- 18423106 TI - Effectiveness of a new tobramycin (0.3%) and dexamethasone (0.05%) formulation in the treatment of experimental Pseudomonas keratitis. AB - OBJECTIVE: To quantitatively determine, in a Pseudomonas keratitis model, the anti-inflammatory and bactericidal properties of a new formulation of tobramycin (0.3%) and dexamethasone (0.05%) that utilizes a xanthan gum vehicle. RESEARCH METHODS: In a randomized and masked fashion, rabbit corneas (n>/=16 eyes per group) were intrastromally injected with 10(3) colony-forming units (CFU) of P. aeruginosa. Eyes were untreated or were administered a single drop every 15 min between 16 and 17 h postinfection (PI) and then a single drop every 30 min between 17 and 22 h PI, a total of 15 drops of either 0.1% dexamethasone and 0.3% tobramycin (TobraDex; Tdex) or a new formulation 0.3% tobramycin and 0.05% dexamethasone with xanthan gum (TobraDex ST; ST). Slit lamp examination scores (SLE+/-SEM) were derived from grading seven parameters at 22 h PI. Rabbits were sacrificed at 23 h PI and the log CFU+/-SEM per cornea was determined. RESULTS: Untreated eyes had SLE scores of 11.11+/-0.43 and had log CFU of 7.27+/-0.06. Eyes treated with Tdex, as compared to the untreated eyes, had significantly lower SLE scores (7.39+/-0.21, p<0.0001) and significantly fewer bacteria (6.32+/ 0.29 log CFU, p=0.0213). Eyes treated with ST had a SLE score (6.56+/-0.19) that was significantly lower than both the untreated eyes (p<0.0001) and the eyes treated with Tdex (p=0.0124). Furthermore, eyes treated with ST had significantly fewer log CFU (5.78+/-0.30) than untreated eyes (p=0.0001) or eyes treated with Tdex (p=0.0434). CONCLUSIONS: The ST formulation with xanthan gum demonstrated statistically superior anti-inflammatory and bactericidal properties as compared to Tdex. LIMITATIONS: Variations in inoculation procedures produced limited eye to-eye differences in the infection. PMID- 18423111 TI - Neuroprotective activities of catalpol on MPP+/MPTP-induced neurotoxicity. AB - Catalpol is an iridoid glycoside, distributed in the roots of Rehmannia glutinosa Libosch. In vitro results showed that preincubation with catalpol (0.5-0.5 mM) for 0.5 hours before and during 48 hour exposure to 0.2 mM MPP+ appeared to be significant protective effect while catalopl was considerably less effective at the doses of 0.001 to 0.01 mM. The addition of catalpol at the dose of 0.05-0.5 mM significantly increased DA and DOPAC to MPP+ treated group. C57bl/6 mice received administration of catalpol for 12 hours before and during the 7 day treatment with MPTP. Such treatment at doses of 15 mg/kg significantly and dramatically blocked tyrosine hydroxylase-positive cell loss in mice. DA turnover in MPTP mice was reversed in the nigrostriatal pathway. In conclusion, data obtained in the above study suggested that catalpol exerted neuroprotective action in vitro and in vivo PD model. PMID- 18423112 TI - Positional sleep apnea in patients with ischemic stroke. AB - BACKGROUND: In non-stroke patients, the severity of sleep apnea (SA) is known to be frequently related to the sleeping position, a condition called positional SA. In the present study, we investigated whether in acute stroke the occurrence of apneas was related to the positioning of patients, and whether a similar finding could be observed after rehabilitation. With the purpose of identifying patients potentially being in need of a SA treatment beyond rehabilitation, we furthermore looked for epidemiologic and clinical parameters being related to persistent SA 6 months after stroke. PATIENTS AND METHODS: Fifty-five acute stroke patients underwent cardiorespiratory polygraphy within 72 hours after onset of neurological symptoms and after 6 months. Apart from the total AHI (AHITOT), the AHI with the patient in supine position and the AHI with the patient in other positions were determined. In all patients, demographic data, NIH-stroke scale score and cumulative vascular risk factors were assessed. RESULTS: In the initial sleep study, 78% of patients had an AHI>or=10/h, of whom 65% fulfilled the criteria of positional SA. On follow-up, the incidence of SA declined to 49% with positional SA being present in 33%. Multivariate logistic regression analysis identified AHITOT on admission [OR=1.07 (1.002-1.13)] and cumulative vascular risk factors [OR=3.48 (1.34-9.05)] as independent predictors of persistent SA 6 months after stroke. CONCLUSION: According to our results, positional SA is a predominant feature in acute stroke and its incidence decreases significantly during the following months. These findings may have implications for SA treatment in patients with acute stroke. PMID- 18423114 TI - Epilepsy surgery in China. AB - Epilepsy surgery began in China from the 1950s to the 1960s, after the People's Republic of China was established (hereinafter as China). However, its rapid growth occurred in recent 10 years with the implement of reform and open policy. The growth is mainly manifested in basic and clinical aspects. As for clinical development, the qualities and quantities of epilepsy surgery have been to an increasing trend gradually. A survey undertaken by the China Association Against Epilepsy (CAAE) in June 2006 at the major six cities in different parts of the country showed a dramatic increase particularly in the years 2004 and 2006. Surgery for temporal lobe epilepsy accounts for half of the operation and it has played an important role in the treatment of the drug resistant epilepsy. With large population and vast geographical spread, it is important to further develop the service, especially for the pediatric epilepsy surgery in China, because the earlier operation for patients with real intractable epilepsy, the better outcome with seizure control and neuropsychologic recovery. PMID- 18423116 TI - [Correlation of sorcin overexpression to multidrug resistance of human gastric cancer cell line SGC7901]. AB - BACKGROUND & OBJECTIVE: Vincristine (VCR)-resistant gastric cancer cell line SGC7901/VCR is a typical multidrug resistant (MDR) cell line with high expression of P-glycoprotein (P-gp). However, verapamil (VRP), the inhibitor of P-gp, can not totally reverse the drug resistance, indicating that additional mechanisms must contribute to the MDR phenotype. Our previous study showed that sorcin, a calcium-binding protein, is significantly up-regulated in SGC7901/VCR cells. This study was to explore the role of sorcin in the development of MDR in human gastric cancer cell line SGC7901. METHODS: The full length sorcin cDNA was isolated by reverse transcription-polymerase chain reaction (RT-PCR). The FLAG sorsin-pcDNA3.1 plasmid was constructed and transfected into SGC7901 cells. The mRNA and protein levels of sorcin in stable clones were detected by RT-PCR and Western blot. The sensitivity of SGC7901 cells to chemotherapeutic drugs was detected using MTT assay. Then sorcin-transfected SGC7901 cells (SGC-F-Sor) were transfected with sorcin antisense oligonucleotides (ASO). The VCR-sensitivity of SGC7901 cells was determined by MTT assay. RESULTS: The full-length sorcin cDNA (616 bp) was amplified by RT-PCR. The FLAG-sorsin-pcDNA3.1 plasmid was constructed successfully. The mRNA and protein levels of sorcin were up-regulated in SGC-F-Sor cells. Overexpression of sorcin produced 8.87 folds of VCR resistance, 6.13 folds of adriamycin (ADM)-resistance, 6.67 folds of taxol resistance, and 2.80 folds of 5-fluorouracil (5-FU)-resistance. However, when SGC F-Sor cells were transfected with sorcin ASO, down-regulation of sorcin expression and increased sensitivity to VCR were observed. CONCLUSIONS: Overexpression of sorcin could induce low level of MDR in SGC7901 cells, indicating that sorcin is associated with MDR of SGC7901 cells. Sorcin maybe be a target of MDR reversal in gastric cancer cells. PMID- 18423117 TI - [Reversal effect of PI-3K/Akt pathway inhibitor LY294002 on multidrug resistance of ovarian cancer cell line A2780/Taxol]. AB - BACKGROUND & OBJECTIVE: Previous studies had showed that phosphatidylinositol 3 kinase (PI-3K) can suppress cell apoptosis. The inhibitor of PI-3K has been used to investigate the mechanisms of PI-3K-induced oncogenesis. This study was to investigate the reversal effect of LY294002, a PI-3K/Akt inhibitor, on paclitaxel resistance of ovarian carcinoma cell line A2780/Taxol. METHODS: A2780/Taxol cells were treated with LY294002. Cell apoptosis was analyzed by flow cytometry (FCM). The 50% inhibition concentration (IC50) of paclitaxel for A2780/Taxol cells was determined by MTT assay. The expression of multidrug resistance 1 (MDR1) mRNA was assessed by reverse transcription-polymerase chain reaction (RT-PCR). The expression of phosphated Akt and P-glycoprotein (P-gp) were detected by Western blot. RESULTS: When treated for 24 h, the apoptosis rate of A2780/Taxol cells was significantly higher in 10 and 50 micromol/L LY294002 groups than in control group [(8.84+/-1.65)% and (20.78+/-2.47)% vs. (1.25+/-0.78)%, P<0.05], the IC50 of paclitaxel for A2780/Taxol cells was decreased significantly (P<0.01) with the highest reverse efficiency of (78.08+/-0.37)%. Moreover, the expression of MDR1 gene, and the phosphorylation of Akt and P-gp in A2780/Taxol cells were decreased. CONCLUSIONS: The activation of PI-3K/Akt pathway plays an important role in paclitaxel-resistance of ovarian carcinoma cells. PI-3K/Akt inhibitor, LY294002 has a reversal effect on the paclitaxel-resistance of A2780/Taxol cells. PMID- 18423118 TI - [Correlation of high expression of HSP27 to multidrug resistance of leukemia cell line K562/VCR]. AB - BACKGROUND & OBJECTIVE: Multidrug resistance (MDR) is a major obstacle preventing effective treatment of leukemia. The mechanisms of MDR in leukemic cells have been broadly explored, but they are still unclear. We used proteomic tools to screen MDR-related proteins in vincristine-resistant leukemia cell line K562/VCR, and analyzed the mechanism of MDR in leukemia. METHODS: Two-dimensional electrophoresis (2-DE) was used to extract total proteins from K562/VCR and K562 cells. The proteins expressed differentially between the two cell lines were identified by matrix-assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF-MS). The antisense oligonucleotide (ASO) of the protein was transfected into K562/VCR cells; mis-sense oligonucleotide (MSO) of the protein was also transfected as control. The expression of the protein was detected by Western blot. Cell survival was detected by MTT assay. Cell apoptosis was detected by flow cytometry (FCM). RESULTS: Heat shock protein 27 (HSP27), a differential expression protein between K562/VCR and K562 cells, was identified. When treated with vincristine, the survival rate of K562/VCR cells was significantly lower in HSP27 ASO group than in HSP27 MSO group (P<0.05). The apoptosis rate of K562/VCR cells was significantly higher in HSP27 ASO group than in HSP27 MSO group (16.37% vs. 3.08%, P<0.05). CONCLUSION: HSP27 is highly expressed in K562/VCR cells, and the suppression of its expression by HSP27 ASO could enhance chemosensitivity of K562/VCR cells to vincristine. PMID- 18423119 TI - [Correlation of tumor vessel function to synergistic antitumor effect of Avastin combined cyclophosphamide]. AB - BACKGROUND & OBJECTIVE: Dysfunction of tumor vessels renders high interstitial pressure, hypoxia and acidosis, causing the barrier of cytotoxic efficacy of chemotherapeutic agents. This study was to observe the dynamic alteration of vessel function in neuroblastoma (NB) after treatment of Avastin, and explore the correlation of tumor vessel function to synergistic antitumor effect of Avastin plus cyclophosphamide (CPM). METHODS: Human NB cells were incubated and transplanted into nude mice to form NB xenografts. Avastin at a dose of 5 mg/kg was administered to the mice through the tail veins. The mice were killed on the 6th hour, 3rd day, 6th day and 9th day after Avastin treatment, separately. Tumor vessel function was tested with fluorescein Hoechst33342 staining. NB-bearing mice were treated with Avastin plus CPM. The synergistic antitumor effects were compared when CPM was administered simultaneously with Avastin (combined regimen I) or at the time the tumor vessel function was mostly improved after Avastin administration (combined regimen II). RESULTS: The tumor vessel function was mostly improved on the 6th day after Avastin treatment. Tumor inhibition rates were 36.4% in Avastin monotherapy group, 38.2% in CPM monotherapy group, 55.9% in combined regimen I group, and 66.8% in combined regimen II group at 3 weeks after treatment. The synergistic antitumor effect was better when CPM was administered on the 6th day after Avastin treatment as compared with it used simultaneously with Avastin (P<0.05). CONCLUSION: The synergistic antitumor effect can be augmented when CPM is administered at the time the tumor vessel function is mostly improved after Avastin treatment. PMID- 18423120 TI - [Inhibitory effects of suramin on growth of transplanted lung adencarcinoma in mice and its mechanisms]. AB - BACKGROUND & OBJECTIVE: Recent study found that suramin could inhibit the growth of malignant tumors, but its in vivo effect on lung adenocarcinoma has seldom been reported. This study was to investigate the inhibitory effects of suramin on the growth and metastasis of transplanted lung adenocarcinoma in mice, and explore the mechanisms. METHODS: Lung adenocarcinoma LA795 cells were transplanted into 32 T739 mice. The tumor-bearing mice were randomized into control group, cisplatin (DDP) group, suramin group, and combination (suramin plus DDP) groupu each group contained 8 mice. Different treatments were served since Day 4 after transplantation. All mice were killed 24 days after transplantation. The metastatic tumor foci on the lung in mice were counted. The occurrence rate of lung metastasis and the inhibition rate of metastatic foci were calculated. The volume and weight of tumors were measured. The expression of epidermal growth factor receptor (EGFR), P-selectin and proliferating cell nuclear antigen (PCNA) in tumor tissues were detected by immunhistochemistry. RESULTS: DDP and suramin used alone or in combination significantly inhibited the growth of LA795 cells in mice (P<0.05), with growth inhibition rates of 34.9%, 23.8% and 57.3%, respectively. The occurrence rate of lung metastasis and the number of metastatic foci on lung surface were significantly lower in suramin group and combination group than in DDP group and control group (P<0.05). The protein levels of EGFR and P-selectin were significantly higher in control group than in DDP group, suramin group, and combination group (157.7+/-6.1 vs. 130.7+/ 5.9, 110.3+/-5.8, and 89.2+/-5.4, P<0.05u 134.5+/-5.7 vs. 117.9+/-5.1, 96.2+/ 5.4, and 78.3+/-4.5, P<0.01). The S phase fraction of tumor cells was significantly higher in control group than in DDP group, suramin group, and combination group [(89.7+/-3.8)% vs. (68.8+/-4.0)%, (65.2+/-4.2)%, and (51.3+/ 4.2)%, P<0.01)]. CONCLUSION: Suramin could inhibit the proliferation and metastasis of LA795 cells in T739 mice through regulating the expression of EGFR, P-selectin and PCNA. PMID- 18423121 TI - [Detecting doxorubicin concentration in KBv200 and KB cell xenografts in nude mice by high-performance liquid chromatography]. AB - BACKGROUND & OBJECTIVE: Doxorubicin (Dox) is one of the most commonly used chemotherapeutic drugs. Drug concentrations in tumor tissue can predict the drugos efficacy better than that in serum do. This study was to detect and compare the concentration of Dox in KBv200 and KB cell xenografts in nude mice by high-performance liquid chromatography (HPLC). METHODS: Tumor models in nude mice were established with KB cells (drug-sensitive) and KBv200 cells (multidrug resistant). Dox was injected via the tail vein. The concentration of Dox in tumor tissue was detected by HPLC at 1, 3 and 5 h after injection. A Hypersil reversed phase BDS C18 column (250 mm x 4.6 mm, ID 5 microm) and mobile phases that were composed of acetonitrile and 0.02 mol/L KH2PO4 (1/2.4, V/V, pH 3.9) at a flow rate of 1.0 mL/min were used for setting a fluorescence detector (excitation wave length was 480 nm; emission wave length was 580 nm). RESULTS: Under the condition of HPLC, the calibration curve of Dox concentration in tumor tissue was linear within a range of 29.3-7 500 ng/g (r=0.9998). The limit of detection in tumor tissue was 14 ng/g. At the concentration of 3 750, 468.8 and 117.2 ng/g, extraction recovery were (99.35+/-7.65)%, (99.79+/-5.73)% and (103.67+/-6.76)%, respectively, method recovery were (91.89+/-7.03)%, (94.94+/-5.18)% and (100.83+/ 5.32)%, respectively. The relative standard deviation (RSD) of the intra-day and inter-day precision were less than 4.2%. At 1, 3, 5 h after Dox injection, the concentrations of Dox were (139.32+/-54.68), (260.00+/-126.11) and (173.26+/ 13.88) ng/g in KBv cell xenografts, respectively, and (385.13+/-42.55), (523.38+/ 138.84) and (460.75+/-86.85) ng/g in KB cell xenografts, respectively. The Dox concentration was significantly higher in KB cell xenografts than in KBv200 cell xenografts at the same time point (P<0.05). CONCLUSION: Detected by HPLC, the concentration of Dox is much lower in multidrug-resistant cell xenografts than in sensitive cell xenografts. PMID- 18423122 TI - [Expression of APRIL in colorectal carcinoma tissues and effects of chemotherapeutic agents on APRIL expression in colorectal carcinoma SW480 cells]. AB - BACKGROUND & OBJECTIVE: A proliferation-inducing ligand (APRIL), a new member of the tumor necrosis factor (TNF) family, can stimulate tumor cell growth and proliferation both in vitro and in vivo. This research was to detect the expression of APRIL in colorectal carcinoma tissues, and to compare the effects of 5-fluorouracil (5-FU) and cisplatin (DDP) on the expression of APRIL in colorectal carcinoma SW480 cells. METHODS: The protein and mRNA levels of APRIL in 56 specimens of human colorectal carcinoma and para-tumor tissues and in SW480 cells were determined by immunohistochemistry and real-time fluorescence quantitative reverse transcription-polymerase chain reaction (FQ-RT-PCR). SW480 cells were treated with 5-FU and DDP at various concentrations for 24 h, 48 h and 72 h. The changes of APRIL mRNA level were analyzed by FQ-RT-PCR. RESULTS: Both positive rate and mRNA level of APRIL were significantly higher in colorectal carcinoma tissues than in para-tumor tissues (76.8% vs. 16.1%, 0.16+/-0.05 vs. 0.71+/-0.08, both P<0.001). The expression of APRIL was strong in SW480 cells. When treated with different concentrations of 5-FU, the mRNA level of APRIL in SW480 cells raised gradually and reached the highest levels at 72 h after treatment (0.85+/-0.10, 0.81+/-0.09, 0.83+/-0.11, and 0.90+/-0.12 at the concentrations of 25, 50, 100 and 200 microg/mL, respectively), which were significantly higher than those in blank control group (P<0.001). When treated with different concentrations of DDP, the mRNA level of APRIL in SW480 cells did not increase when compared with that in control group (P>0.05). After 72-hour treatment, the mRNA level of APRIL in SW480 cells was significantly lower in 10 microg/mL and 20 microg/mL DDP groups than in blank control group (0.44+/-0.05 and 0.40+/-0.07 vs. 0.57+/-0.06, P<0.05). CONCLUSIONS: APRIL may promote the development of colorectal carcinoma. When chemotherapy is conducted to treat colorectal carcinoma, especially when 5-FU is included in the regimen, anti-APRIL therapy might be an important assistant treatment to counter the impact of APRIL caused by antitumor drugs. PMID- 18423123 TI - [Synergistic lethal effect of mDRA-6 and nimesulide on human hepatocellular cancer cell line SMMC-7721]. AB - BACKGROUND & OBJECTIVE: Both mDRA-6, a monoclonal antibody of death receptor 5 (DR5) in human cells prepared by our key laboratory, and nimesulide, a specific cyclooxygenase-2 (COX-2) inhibitor, can induce apoptosis of some malignant tumor cells. This study was to investigate the lethal effects of mDRA-6 and nimesulide on human hepatocellular cancer cell line SMMC-7721, and explore the possible mechanism. METHODS: The expression of DR5 on SMMC-7721 cells was detected by flow cytometry (FCM). SMMC-7721 cells were treated with mDRA-6 and nimesulide alone or in combination. Cell morphology was observed under microscope with Hoechst33258 staining. Cytotoxicity was examined by MTT assay. Cell apoptosis was detected by FCM. RESULTS: The positive rate of DR5 on SMMC-7721 cells was 95.0%. The apoptosis of SMMC-7721 cells could be induced by both mDRA-6 and nimesulide: the apoptosis rates were 10.5% when treated with 25 ng/mL mDRA-6 for 12 h, 35.0% when treated with 1 600 ng/mL mDRA-6, 5.0% when treated with 200 micromol/L nimesulide, and 34.0% when treated with 800 micromol/L nimesulide. The combination of mDRA-6 and nimesulide exhibited synergistic effect on the apoptosis of SMMC-7721 cells (q=1.23): the apoptosis rates were 31.2% when treated with 200 micromol/L nimesulide and 25 ng/mL mDRA-6 for 12 h, and 91.1% when treated with 200 micromol/L nimesulide and 1 600 ng/mL mDRA-6 for 12 h. CONCLUSIONS: Both mDRA-6 and nimesulide can induce the apoptosis of SMMC-7721 cells. The combination of mDRA-6 and nimesulide exhibits synergistic lethal effect on SMMC-7721 cells. PMID- 18423124 TI - [Screening concurrent chemoradiotherapy sensitivity-associated proteins in intermediate stage and advanced cervical carcinoma]. AB - BACKGROUND & OBJECTIVE: Concurrent chemoradiotherapy is a new therapy for intermediate stage and advanced cervical carcinoma, but no valid index for prediction of concurrent chemoradiotherapy sensitivity is available. This study was to screen concurrent chemoradiotherapy sensitivity-associated proteins in intermediate stage and advanced cervical carcinoma. METHODS: Biopsy samples of 10 cervical carcinoma patients were collected before treatment. According to their responses to concurrent chemoradiotherapy (WHO standard), the patients were classified into high sensitivity (HS) group (5 patients) and low sensitivity (LS) group (5 patients). Total protein were extracted from the biopsy samples. Differential proteins were detected by two-dimensional gel electrophoresis (2-DE) and confirmed by matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). Two differentially expressed proteins were further detected by immunohistochemistry in 95 specimens of cervical carcinoma, including 60 high sensitive cases and 35 low sensitive cases. RESULTS: Nineteen differentially expressed proteins were identified: 9 were highly expressed and 10 were lowly expressed in high sensitive group as compared with those in low sensitive group. According to immunohistochemical results, the expression intensity of heat shock protein 70 (HSP70) was higher and that of S100A9 protein was lower in HS group than in LS group; the positive rate of S100A9 was significantly higher and that of HSP70 was significantly lower in HS group than in LS group (88.3% vs. 28.6%, chi2=35.34, P<0.001; 21.7% vs. 85.7%, chi2=36.59, P<0.001). CONCLUSIONS: Differentially expressed proteins that related to concurrent chemoradiotherapy sensitivity of cervical carcinoma are identified. They may be candidate biomarkers for prediction of concurrent chemoradiotherapy sensitivity. PMID- 18423125 TI - [Effects of cetuximab combined erlotinib on proliferation of human hepatocellular carcinoma cell lines HepG2 and Bel-7402]. AB - BACKGROUND & OBJECTIVE: Several researches have reported that epidermal growth factor receptor (EGFR) is expressed frequently in hepatocellular carcinoma (HCC) and paratumor tissues, most likely to contribute to the biological characteristics of HCC, including invasion, metastasis, and resistance to chemotherapy and radiotherapy. The single agent of cetuximab and erlotinib can inhibit the proliferation of HCC cells in vivo and in vitro. This study was to investigate the effects of cetuximab combined erlotinib on the proliferation of HCC cells, and explore the molecular mechanism. METHODS: Increasing concentrations of cetuximab (5-500 mg/mL) and erlotinib (2.5-250 micromol/L) alone or in combination were administrated to HCC cell lines HepG2 and Bel-7402. The inhibitory effects of the drugs on cell proliferation at different time points were observed; the combination index (CI) of these two agents was calculated. The expression of key enzymes in EGFR signaling transduction pathway in HepG2 and Bel-7402 cells after different treatments was detected by Western blot. RESULTS: The single agent of cetuximab and erlotinib inhibited the proliferation of HCC cells in a time-and dose-dependent manner; after 72-hour treatment, the proliferation inhibition rates of HepG2 cells were (43.1+/-1.9)% and (83.4+/-1.3)%, and those of Bel-7402 cells were (35.1+/-2.6)% and (73.9+/ 1.2)%. With maximal inhibitory effects of cetuximab combined erlotinib, the proliferation inhibition rates of HepG2 and Bel-7402 cells were (91.1+/-1.0)% and (84.6+/-1.1)%, respectively. The CIs of different concentrations of these two agents at different time points were all less than 1, suggested that they have obvious synergistic activity. The expression of activated key enzymes (p-EGFR, p ERK1/2, p-AKT) in EGFR signaling transduction pathway in HepG2 and Bel-7402 cells were down-regulated more obviously when these two agents were administrated in combination. CONCLUSIONS: The single agent of cetuximab and erlotinib can inhibit the proliferation of HepG2 and Bel-7402 cells through down-regulating activated key enzymes in EGFR signaling transduction pathway, and they have obvious synergistic activity. The combined EGFR-targeting therapy on HCC xenografts in vivo and on HCC patients is worth of further exploration. PMID- 18423126 TI - [Efficacy of erlotinib on advanced non-small cell lung cancer]. AB - BACKGROUND & OBJECTIVE: Erlotinib is a selective inhibitor of epidermal growth factor receptor (EGFR) tyrosine kinase, and has been used in treating advanced non-small cell lung cancer (NSCLC). This study was to evaluate the efficacy of erlotinib on advanced NSCLC, and observe the adverse events. METHODS: An open labeled, expanded access program (EAP) was conducted on 44 pathologically confirmed advanced NSCLC patients who had received at least one regimen. Erlotinib (150 mg) was orally administered daily till disease progression or intolerable adverse events developed. The efficacy was evaluated according to RECIS criteria; the adverse events were evaluated according to NCI criteria. RESULTS: In the 44 patients, the objective response rate was 27.3%, and the disease control rate was 65.9%u the median progression-free survival time was 4.5 months (0.9-8.1 months), and the median survival time was 13.7 months (9.2-18.2 months). Adverse events were generally mild (grade I or II), including skin rash (81.8%) and diarrhea (56.8%). One (2.3%) patient developed grade III elevation of serum glutamate pyruvate transaminase (SGPT). No grade IV drug-related adverse event occurred. CONCLUSION: Erlotinib is effective and safe for locally advanced or metastatic NSCLC patients who have failed previous chemotherapy. PMID- 18423128 TI - [Meta analysis of clinical effectiveness of neoadjuvant chemotherapy for gastric cancer]. AB - BACKGROUND & OBJECTIVE: Neoadjuvant chemotherapy has gained increasing attention as a treatment for gastric cancer since Wilke first reported its application to the treatment of gastric cancer in 1989. However, its value in treating gastric cancer remains controversial. This research was to assess the efficacy of neoadjuvant chemotherapy on gastric cancer through a Meta analysis of the randomized controlled trials published worldwide in both English and Chinese. METHODS: Cochrane systematic evaluation was used to search through Cochrane libraries of clinical comparative trials, PubMed, Embase, Chinese Biomedical Literature Database (CBM), Chinese Scientific Journal Full-text Database (CSJD) and Chinese Journal Full-text Database (CJFD), aided with manual retrieval and other retrievals. The quality of the assessment was independently evaluated and cross-checked by two evaluators, and the results of homogeneous studies were analyzed with RevMan4.2.10 software. RESULTS: Five randomized controlled trials involved a total of 838 patients were studied. Of the 5 trials, 2 were performed in Japan, 1 in the Netherlands, 1 in the United Kingdom, 1 in China. Of the 838 patients, 373 were treated with neoadjuvant chemotherapy and 465 were treated with surgery alone. Among the above 5 studies, one used blind method and one described random allocation concealment method. No statistical differences were found in the resection rate, cure rate, 1-and 5-year survival rates between neoadjuvant chemotherapy group and surgery group [odds ratio (OR)=1.09, 95% confidence interval (CI)=0.67-1.77 for resection rate; OR=1.25, 95% CI=0.85-1.84 for cure rate; OR=1.61, 95% CI=0.90-2.90 for 1-year survival rate; OR=1.13, 95% CI=0.83-1.53 for 5-year survival rate]. CONCLUSIONS: The efficacy of neoadjuvant chemotherapy on gastric cancer is not better than that of surgery alone. Therefore, neoadjuvant chemotherapy should not be recommended as a regular treatment for gastric cancer before obtaining evidences of its certain efficacy on gastric cancer. PMID- 18423127 TI - [Responses of advanced esophageal cancer to chemotherapy and prognostic factors: a report of 138 cases]. AB - BACKGROUND & OBJECTIVE: The prognosis of advanced esophageal cancer is poor. There are no definite prognostic factors and standard regimens for these patients. This study was to analyze the responses of advanced esophageal cancer to chemotherapy, and explore its probable prognostic factors. METHODS: Clinical data of 138 naive patients with histologically or pathologically confirmed advanced esophageal cancer, treated from Dec. 1984 to Apr. 2006 in Cancer Hospital of Chinese Academy of Medical Sciences, were analyzed using Chi-square test, Kaplan-Meier method, and log-rank test. Of the 138 patients, 68 were treated with taxol or gemcitabine or oxaliplatin (new drug group), including 64 (94.1%) treated with cisplatin; 70 were treated without taxol, gemcitabine and oxaliplatin (conventional drug group), including 48(68.6%) treated with cisplatin. RESULTS: The response rate of 138 patients was 47.8%. The median time to progression (TTP) was 4 months; the median survival time was 10 months. The response rate was significantly higher in new drug group than in conventional drug group (58.8% vs. 37.1%, P=0.011). Univariate analysis indicated that age, hemoglobin (HB) level before treatment, chemotherapy cycles, short-tem efficacy, TTP and therapeutic methods were significant prognostic factors. Cox multivariate regression analysis showed that TTP, therapeutic methods and HB level before treatment were independent prognostic factors. CONCLUSIONS: Taxol or gemcitabine combined cisplatin has certain effect on advanced esophageal cancer. TTP, therapeutic methods and HB level before treatment are independent prognostic factors of this disease. PMID- 18423129 TI - [FOLFOX4 regimen versus DP(O)F regimen for advanced gastric cancer]. AB - BACKGROUND & OBJECTIVE: Currently, there is no standard regimen for advanced gastric cancer. FOLFOX4 regimen [oxaliplatin(L-OHP) with 5-fluorouracil (5-FU)] and DP(O)F regimen [docetaxel (TXT), oxaliplatin (L-OHP)/cisplatin (DDP) with 5 FU] are usually used in treating gastric cancer with satisfactory efficacy. This study was to compare the efficacy, time to disease progression (TTP), overall survival (OS) and toxicity of the two regimens for advanced gastric cancer. METHODS: Clinical data of 70 chemotherapy-naive patients with advanced gastric cancer were analyzed. Of the 70 patients, 34 were treated with FOLFOX4 regimen, 36 were treated with DP(O)F regimen. The patients in FOLFOX4 group received intravenous infusion of L-OHP (85 mg/m(2)) at Day 1, bolus injection of 5-FU (400 mg/m(2)) at Days 1-2, and continuous intravenous infusion of 5-FU (600 mg/m(2)) for 22 h at Days 1-2; 14 days as one cycle. The patients in DP(O)F group received administration of TXT [20 mg x (m2 x w)(-1)], intravenous infusion of DDP (40 mg/m(2)) at Days 2-3 or L-OHP (100 mg/m(2)) at Day 2, and intravenous infusion of 5-FU (500 mg/m(2)) at Days 1-5; 21 days as one cycle. RESULTS: The objective response rates were 45.4% in FOLFOX4 group and 52.8% in DP(O)F group (P=0.628). The median TTP was 5.27 months in FOLFOX4 group and 4.70 months in DP(O)F group (P=0.848). The median survival time was 8.97 months in FOLFOX4 group and 12.17 months in DP(O)F group (P=0.095). The most frequent adverse events were nausea, vomit and hematologic toxicities. The occurrence rates of grade III-IV leukopenia and neutropenia were significantly lower in FOLFOX4 group than in DP(O)F group (11.8% vs. 36.1%, P=0.025; 17.6% vs. 41.7%, P=0.038). One patient in FOLFOX4 group died of liver function failure. CONCLUSION: Both FOLFOX4 and DP(O)F regimens are effective in treating advanced gastric cancer. The hematologic toxicities of DP(O)F regimen are worse than those of FOLFOX4 regimen. PMID- 18423130 TI - [Influence of intravenous chemotherapy on cellular immunity in patients with advanced digestive tract cancer]. AB - BACKGROUND & OBJECTIVE: Cancer patients have a poor immune response and chemotherapy could deteriorate their immune system further. Reasonable immune therapy is an important adjuvant remedy for tumors. This study was to monitor the changes of T-cell phenotypes in peripheral blood and interleukin-2 (IL-2) concentration in plasma in digestive tract cancer patients before and after chemotherapy. METHODS: The proportions of CD3+, CD4+, CD8+, CD4+CD28+, CD8+CD28+ and CD4+CD25+ T cells in peripheral blood of 104 patients with advanced digestive tract cancer, hospitalized from Sep. 2005 to Apr. 2006, were detected by flow cytometry (FCM). The concentration of IL-2 in plasma was measured by ELISA. RESULTS: The proportions of CD4+, CD4+CD28+, CD8+CD28+, CD4+CD25+ T cells and ratio of CD4/CD8 were (36.52+/-3.85)%, (32.87+/-4.98)%, (6.87+/-3.11)%, (9.68+/ 3.42)% and 0.98+/-0.17 in digestive tract cancer patients, and (45.23+/-9.20)%, (40.12+/-5.85)%, (15.8+/-4.50)%, (5.67+/-2.90)% and 1.43+/-0.12 in healthy subjects. In the patients with response to chemotherapy, the proportions of CD4+CD28+ and CD8+CD28+ T cells and ratio of CD4/CD8 were (22.93+/-3.98)%, (7.08+/-1.23)% and 0.90+/-0.22 before chemotherapy, and (28.25+/-4.03)%, (12.10+/ 3.45)% and 1.24+/-0.22 at 3 weeks after chemotherapy. In the patients with no response to chemotherapy, the proportions of CD4+CD28+, CD8+CD28+ and CD4+CD25+ T cells were (24.08+/-4.02)%, (6.35+/-1.23)% and (8.20+/-2.34)% before chemotherapy, and (16.45+/-3.27)%, (3.20+/-0.82)% and (20.34+/-3.69)% at 3 weeks after chemotherapy. CONCLUSIONS: The immunosuppression of digestive tract cancer patients would be enhanced early (about 1-2 weeks) after intravenous chemotherapy. The immunity of the patients with response to chemotherapy would be improved at 3 weeks after chemotherapy; while the immunity of the patients with no response to chemotherapy would not change, or even be suppressed. PMID- 18423131 TI - [Clinical analysis of 69 cases of Burkitt's lymphoma]. AB - BACKGROUND & OBJECTIVE: Burkitt's lymphoma is a kind of highly aggressive B-cell lymphoma. Its clinical characteristics are different between the endemic areas in Africa and the sporadic areas in America and Europe. There is no large-scale report concerning Burkitt's lymphoma in China yet. This study was to summarize the characteristics of Burkitt's lymphoma in China. METHODS: Clinical data of 69 Burkitt's lymphoma patients, treated from May 1985 to May 2007 in Cancer Center of Sun Yat-sen University, were analyzed. RESULTS: Of the 69 patients, 44 were men and 25 were women, with a median age of 7 (range, 2-72); 5 were at stage I, 9 at stage II, 21 at stage III, and 34 at stage IV, advanced stage (stages III and IV) accounted for 55 (79.7%) patients. Abdomen (63.8%), cervical lymph nodes (68.1%) and faciomaxillary-oropharynx (34.8%) were the most common involved sites. Bone marrow (21.9%) and central nervous system (17.4%) could also be involved. B symptoms were found in 34 patients. Serum lactate dehydrogenase (LDH) level was elevated in 42 of 58 patients, while serum uric acid level was elevated in 13 of 56 patients. Hepatitis B virus (HBV) infection was found in 6 of 57 patients, Epstain-Barr virus (EBV) infection in 7 of 13 patients, human immunodeficiency virus (HIV) infection in 0 of 51 patients. Short-term and high intensive chemotherapy with central nervous system prophylaxis could improve the prognosis. CONCLUSION: The clinical characteristics of these 69 Burkitt's lymphoma patients are much similar to those from sporadic areas, but the median age is lower, and the most common involved sites are cervical lymph nodes, abdomen and faciomaxillary-oropharynx. PMID- 18423132 TI - [Efficacy of bortezomib combined dexamethasone in 24 patients with multiple myeloma]. AB - BACKGROUND & OBJECTIVE: Bortezomib, a potent and reversible proteasome inhibitor, induces apoptosis of myeloma cells, resulting in durable responses in patients with multiple myeloma (MM). This study was to explore the medical effects and side effects of bortezomib combined dexamethasone in treating newly diagnosed and relapsed or refractory MM, and evaluate the safety of this regimen in the patients with renal impairment. METHODS: Twenty-four MM patients were treated with bortezomib and dexamethasone in a 21-day cycle. The patients received a median of 3 cycles (range, 1-8 cycles) of the treatment. Response to bortezomib was evaluated according to the criteria of the European Group for Blood and Marrow Transplantation (EBMT); adverse events were graded according to the National Cancer Institute Common Toxicity Criteria. RESULTS: During the follow-up with a median of 4 months, 19 (79.2%) patients responded to the treatment. The complete remission (CR) rate was significantly higher in the patients of light chain type than in those of non-light-chain type (57.1% vs. 5.9%, P=0.014). The response rates of the patients with and without renal impairment were similar (100% vs. 70.6%, P=0.272), and the renal functions were ameliorated in the patients with renal impairment during chemotherapy. Grade III-IV adverse events, including leucocytopenia (8.3%), thrombocytopenia (33.3%), diarrhea (8.3%) and debility (4.2%), could be relieved by symptomatic treatment or delayed chemotherapy. CONCLUSIONS: The combination of bortezomib and dexamethasone shows obvious effects on MM, especially in the patients with light-chain type. The adverse events can be tolerant in most patients, and this regimen is also safe in the patients with renal impairment. PMID- 18423133 TI - [Efficacy of DACE regimen on relapsed and refractory non-Hodgkin's lymphoma]. AB - BACKGROUND & OBJECTIVE: There is no standard salvage regimen for patients with relapsed and refractory non-Hodgkin's lymphoma (NHL) at present. This study was to evaluate the efficacy of DACE regimen, as a salvage regimen, on relapsed and refractory NHL, and observe the adverse events. METHODS: From May 2001 to May 2006, 61 patients with relapsed and refractory NHL received chemotherapy of DACE regimen: intravenous infusion of cisplatin (20 mg/m(2)) on Days 1-5, etoposide (100 mg) on Days 1-5, cytarabine (150 mg) on Days 1-3, and dexamethone (15 mg/m(2)) on Days 1-5; repeated every 3 weeks. The efficacy and adverse events were evaluated according to the World Health Organization criteria. RESULTS: The overall response rates were 63.9% after two cycles of chemotherapy and 72.1% after four cycles. The median remission time lasted 4.7 months (1-58 months). The 1-year survival rate was 29.5%u the 2-year survival rate was 21.3%. Myelosuppression was the major adverse event: the occurrence rate of grade III-IV myelosuppression was 49.1%u it was well tolerated. CONCLUSION: DACE regimen is an effective salvage regimen in treating patients with relapsed and refractory NHL. PMID- 18423134 TI - [Treatment outcome of primary central nervous system germ cell tumors after combined therapy: a report of 23 cases]. AB - BACKGROUND & OBJECTIVE: Primary central nervous system (CNS) germ cell tumors (GCTs) are rare malignant neoplasms with various histological types. Excluding pure germinoma and mature teratoma, other types carry a poor prognosis. Previous investigations focused on combined modality treatment including chemotherapy to improve survival. This study was to analyze the efficacy and toxicity of chemotherapy combined with surgery and/or radiotherapy on CNS GCTs. METHODS: A total of 23 patients with CNS GCTs were treated in Cancer Center of Sun Yat-sen University from May 2002 to Jun. 2006. The median age at diagnosis was 16 years. Alpha-fetoprotein (AFP) and/or beta-human chorionic gonadotropin (beta-HCG) levels were elevated in 19 patients (82.6%). All patients were treated with chemotherapy of PEB regimen combined with surgery and/or radiotherapy. PEB regimen was administered every 3 weeks with 20 mg/m(2) cisplatin (DDP) at Days 1 5 or 80-100 mg/m(2) at Day 1, 60-100 mg/m(2) etoposide (VP-16) or teniposide (VM 26) at Days 1-5, 10 mg/m(2) bleomycin (BLM) at Days 1 and 5. RESULTS: Of the 23 patients, 17 newly diagnosed patients received induction chemotherapy followed by radiotherapy or surgery/radiotherapy followed by adjuvant chemotherapy; 6 recurrent patients received salvage chemotherapy, of which 3 patients with disseminated tumor received salvage chemotherapy followed by craniospinal irradiation. The 23 patients completed a total of 61 cycles of PEB regimen with a median of 3 cycles. Chemotherapy alone gained a response rate of 87.0% and a complete remission rate of 30.4%. Craniospinal irradiation was performed in 14 patients and focal irradiation in 8 patients. One patient did not receive irradiation. Incomplete tumor resection or biopsy was performed in 13 patients. Fourteen patients (60.9%) were alive without evidence of diseases after combined modality treatment. With a median follow-up of 24 months, the 2-year survival rate was 67.4%. Main adverse events were late irradiation injury including aplastic anemia and hypothalamus syndrome (1 case), irradiation encephalopathy (1 case), and hypopituitarism (2 cases). CONCLUSIONS: The combined modality treatment including PEB regimen is highly effective in treating CNS GCTs patients, especially in the patients with elevated tumor markers. However, the long-term toxicities which related with craniospinal irradiation should not be ignored. PMID- 18423135 TI - [Anti-angiogenesis targeting drugs: a review]. AB - The dependence of tumor growth and metastasis on blood vessels makes tumor angiogenesis a rational target for cancer therapy. One of the key mediators of angiogenesis is vascular endothelial growth factor (VEGF), which is an appealing target for anticancer therapy. The development of anti-VEGF/VEGFR agents and the latest clinical data are reviewed, including bevacizumab as a monoclonal antibody to VEGF, sunitinib and sorafenib as VEGFR tyrosine kinase inhibitors, and IMC 1C11 as VEGFR monoclonal antibody. PMID- 18423137 TI - [Regulations of and advances in liver transplantation]. PMID- 18423138 TI - [Opinions about the 2008 Asian Pacific Association for the Study of the Liver guidelines for chronic hepatitis B management]. PMID- 18423139 TI - [The optimal time for liver transplantation]. PMID- 18423140 TI - [Current status of adult-to-adult living donor liver transplantation]. PMID- 18423141 TI - [Advances in using marginal donors in liver transplantation]. PMID- 18423142 TI - [Monitoring immunosuppressive agents in liver transplantation patients]. PMID- 18423143 TI - [Outcomes of liver transplantation in different liver diseases and their impact factors]. PMID- 18423144 TI - [Management of biliary complications in liver transplantation]. PMID- 18423145 TI - [Impact factors on reinfection after routine treatment and liver transplantation in hepatitis C patients]. PMID- 18423146 TI - [Enhancing active immunity against hepatitis B virus by HBV vaccine immunization in patients with HBV-related end-stage liver diseases treated with liver transplantation]. AB - OBJECTIVES: To study the active immunity response of liver transplant patients for HBV-related diseases after hepatitis B virus (HBV) vaccine immunization and to investigate the factors that influence the effectiveness of the vaccination in order to find measures to increase its success. METHODS: Thirteen patients who had liver transplants because of HBV-related end-stage liver diseases received hepatitis B virus immunoglobulin and lamivudine for an average of 38 months (range 27-77 months). They received double intramuscular doses (40 microg) of a recombinant vaccine at months 0, 1, 2 and 6. The anti-HBs titers were tested regularly at months 1, 2, 3, 6 and 7. RESULTS: Seven of the 13 patients (53.8%) developed higher serum titers of anti-HBs compared with their titers prior to the vaccinations, 2 patients of the 13 (15.4%) developed an increase by 100 U/L and in 4 patients (30.8%) their base levels were doubled. Those responding patients were followed-up for another 8 months after the fourth vaccination, and only 1 patient among them had a decrease of the anti-HBs titers below the level prior to the vaccination. CONCLUSION: Hepatitis B vaccine immunization can be used to enhance the active immunity against HBV in patients who had liver transplants for HBV-related diseases. PMID- 18423147 TI - [A study on mutations of the overlapping hepatitis B virus surface and polymerase gene in patients with HBV reinfection after liver transplantations]. AB - OBJECTIVE: To investigate the influence of combined hepatitis B immune globulin (HBIG) and lamivudine (LMV) treatment on hepatitis B virus (HBV) surface antigen and polymerase overlapping gene mutations in HBV reinfected liver transplant recipients. METHODS: From June 2002 to December 2003, 320 patients who underwent liver transplantations due to HBV-related end-stage liver diseases were followed up for 1.5 to 3 years postoperatively. Fourteen patients developed HBV reinfection. They had LMV before their liver transplantations and had LMV and HBIG after the transplantations to prevent HBV infections. Their serum levels of HBV DNA were measured by polymerase chain reaction. Gene sequencing method was used to analyze HBV genotype and mutations of the S gene. Micro-particle enzyme immunoassay was used to measure the serum concentration of HBIG. RESULTS: (1) There was no obvious difference in the number of amino acid mutation sites in S and P regions before and after the transplantations. (2) The HBV genotypes were B type (n=2) and C-type (n=12) in the reinfected group before the transplantations, and genotypes after the transplantations remained the same. (3) HBIG concentrations were 0 U/L in 7 patients, less than 100 U/L in 5 patients, and more than 100 U/L in 2 patients. Mutations were detected as I126S, T131N, S143T and G145R in 'a' determinant and L110F, I113S, T160K in up- or down-stream of 'a' determinant. (4) Mutations in S gene caused missense mutation in the surface antigen region. These mutations also caused corresponding missense mutations in the polymerase region. The missense mutation in the polymerase region involved lamivudine mutation sites and other mutation sites. CONCLUSION: Immunosuppressant therapy has no obvious influence on the numbers of mutations, but it can influence the sites of the mutations. Besides 'a' determinant mutations, there exist mutations in up- or down-streams of 'a' determinant and they may cause HBV reinfection. PMID- 18423148 TI - [Living-related liver transplantation for cavernous transformation of portal vein: a clinical study of 3 cases]. AB - OBJECTIVE: To review the outcomes of living-related liver transplantation (LRLT) in treating 3 cases of cavernous transformation of portal vein (CTPV) with severe portal hypertension. METHODS: Three children (two boys and one girl) were presented to our hospital with recurring esophageal variceal bleeding, decompensating ascites, splenomegaly and refractory anemia. CTPV was confirmed by intravenous computed tomographic portography using a helical computed tomography scanner and 3-dimensional image reconstruction. LRLT were performed in these 3 patients from July 2006 to January 2007. The evaluation of the outcomes was made by referring to their clinical features and laboratory and imaging examination findings. RESULTS: Although one patient died from early graft thrombosis, the other two patients showed excellent prognoses. They lived and stayed well during a follow-up period of 12-14 months. Following the transplantations, there had been no esophageal variceal hemorrhage, the ascites disappeared and the portal hypertension vanished. Their hemoglobin, blood platelets count, and serum albumin reached normal values. CONCLUSION: LRLT is an effective procedure in treating CTPV with severe portal hypertension. The reconstruction of the portal vein is the difficult part of the LRLT procedure. PMID- 18423149 TI - [Using 1H-MRS to evaluate the effects of warm ischemia reperfusion injury on the regeneration of hepatic cells in orthotopic transplanted livers in rats]. AB - OBJECTIVE: To investigate the prognostic value of proton magnetic resonance spectroscopy (1H-MRS) in the study of warm ischemia reperfusion injury to the regeneration of hepatic cells of the livers following their orthotopic transplantation in rats. METHODS: A rat orthotopic liver transplantation (OLT) model with warm ischemia, the experimental group, was established and the same was done with a control group but without warm ischemia of the livers. They were studied at 6 time points (6 hours, 1, 3, 7, 14, and 30 days after OLT). All rats took axial T1 weighted and T2 weighted imaging scans and 1H MR spectroscopies. RESULTS: The positive rate of proliferating cell nuclear antigen (PCNA) and the mean peak choline/water ratio in the experimental group were significantly higher than those in the control group and the peak choline /water ratio had a positive correlation with the positive rate of PCNA. Serum ALT and AST increased significantly after OLT, especially during the 6 hour to 3day period. The levels of ALT and AST were markedly higher in the experimental group compared to the control group. CONCLUSION: Warm ischemia reperfusion injury of OLT has a significant effect on the regeneration of hepatic cells, and the choline peak of 1H-MRS can be used to evaluate the regeneration of hepatic cells non-invasively. PMID- 18423150 TI - [Responses of peginterferon-alpha 2a antiviral therapy in chronic hepatitis B patients]. AB - OBJECTIVE: To study the responses of peginterferon-alpha 2a antiviral therapy in chronic hepatitis B (CHB) patients. METHODS: One hundred two CHB patients with their serum ALT values higher than 2x the upper limit of the normal (ULN) were divided into a HBeAg-positive and a HBeAg-negative group. All patients were treated with peginterferon-alpha 2a by subcutaneous injection (180 microgram once weekly). After treatment for 6 months, patients without a defined therapeutic response were dropped from the treatment group; the others completed a 12 month therapy. The sustained response and the antiviral effect of the treatment were assessed at the end of the therapy. To investigate the possible impact factors of the response to peginterferon-alpha 2a, we studied the therapeutic response of patients with different serum ALT levels, inflammation grades of liver histology, stages of fibrosis, and HBV viral load levels. RESULTS: (1) There was no statistical difference of the rates of response at the end of treatment and 6, 12, 18, 24 and 30 months after the cessation of therapy between the HBeAg positive and the HBeAg-negative groups. (2) In the HBeAg-positive group, the rates of response of patients with serum ALT values>3*ULN were significantly higher than those with serum ALT values less than 3*ULN (x2=4.40). However, no statistical difference of serum ALT levels was found in the HBeAg-negative group. (3) In both HBeAg-positive and HBeAg-negative groups, no difference was revealed in the rates of response among patients with different levels of HBV viral loads. (4) In the HBeAg-positive group, patients with more severe liver inflammation histologically (G3 and G4) had significantly higher response rates than those with milder inflammation (G1 and G2) (x2=4.19), but no similar statistical differences were found in the HBeAg-negative group. Moreover, there was no difference in the rates of response among patients in different stages of liver fibrosis in both HBeAg-positive and HBeAg-negative groups. CONCLUSIONS: Similar rates of response and sustained virological response to the peginterferon-alpha 2a treatment can be achieved in both HBeAg-positive and HBeAg-negative patients. Hepatic fibrosis is not a predictor of poor therapeutic response. For HBeAg positive patients, more severe liver inflammation identified with liver biopsies (G3 or G4) and high serum ALT values (more than 3*ULN) can be considered as predictors of a good therapeutic response. PMID- 18423151 TI - [Changes of plasma cytokines in patients with severe hepatitis treated with a probiotic preparation]. AB - OBJECTIVE: To study the changes of plasma cytokines in patients with severe hepatitis treated with a probiotic preparation. METHODS: One hundred twelve patients with severe hepatitis treated with a basic regimen were randomly divided into a probiotic preparation treatment group and a control group. Their plasma cytokines such as tumor necrosis factor alpha, interleukin-2, interleukin-6, interleukin-10, were determined by conventional techniques. RESULTS: Clinical symptoms of 84.5% of the treatment group were obviously improved. The treatment effectiveness in this group was better than that in the control group (X2=8.888). It showed that liver functions were significantly improved compared to those of the control group . After the probiotic preparation therapy, there were significant decreases in the concentrations of TNFa and IL-6. TNFa and IL-6 in the treatment group and in the control group were (109.4+/-14.7) pg/ml vs (128.7+/-18.8) pg/ml and (84.3+/-20.1) pg/ml vs (109.1+/-18.7) pg/ml respectively. However, the concentrations of IL-10 and IL-2 in the treatment group increased obviously: IL-2 was (59.8+/-12.2) pg/ml vs (47.1+/-6.7) pg/ml and IL-10 was (30.6+/-6.6) pg/ml vs (22.5+/-6.1) pg/ml. CONCLUSION: The probiotic preparation treatment effectively reduces the TNFa and IL-6 and increases the concentrations of IL-10 and IL-2 in the blood of the patients with severe hepatitis. PMID- 18423152 TI - [Screening and cloning gene of a hepatocyte protein interacting with HCV NS5ATP4A protein]. AB - OBJECTIVE: To screen and clone hepatocyte protein interacting with hepatitis C virus NS5ATP4A protein for studying its biological functions. METHODS: Bait plasmids of hepatitis C virus NS5ATP4A were constructed. After verifying that hepatitis C virus NS5ATP4A protein could be steadily expressed in AH109 yeast strain, yeast-two hybrid assay was performed by mating AH109 with Y187 which pre transformed with liver cDNA library plasmids pACT2, and the diploidy yeast cells were plated on quadruple dropout (QDO) medium and assayed for X-a-gal activity. Nineteen yeast colonies which grew on QDO and had a-gal activity were obtained, and then the library plasmids were extracted and sequenced. RESULTS: Seven genes were screened out and one of them was a formerly unknown gene. They were associated with RNA synthesis, protein translation, cell cycling and tumor immunity. CONCLUSION: NS5ATP4A binding proteins were successfully screened, which offers new clues for further studying the signal transduction pathway of NS5ATP4A and the pathogenic mechanism of HCV. PMID- 18423153 TI - [Adenovirus vector encoding human KDR elicits immunity against hepatocellular carcinomas in mice]. AB - OBJECTIVE: To investigate the effect of adenovirus vector encoding human vascular endothelial growth factor receptor-2 (hVEGFR-2 or hKDR) on breaking the immune tolerance and inducing immunity against murine hepatocellular carcinomas. METHODS: Human and mouse KDR cDNA were cloned from human umbilical vein endothelial cells (HUVEC) and C57BL/6 mouse embryo cells respectively using RT PCR, and then Ad hKDR and Ad mKDR were constructed. Seven days after immunization of the mice with Ad hKDR or Ad mKDR, an analysis of cytotoxic activity of antigen specific cytotoxic T lymphocytes (CTL) was made by lactate dehydrogenase (LDH) release assay, in which splenocytes of the immunized mice acted as effectors and Hepa 1-6/mKDR cells as the targets. In addition, the survival of the mice immunized with Hepa 1-6 hepatoma cells was checked. RESULTS: Seven days after immunization, the 6 h killing activities of CTL elicited by the Ad hKDR were 84.3%+/-6.7%, 71.5%+/-5.2%, and 44.6%+/-4.7% at the ratio of the effectors:targets (E:T) of 100:1, 50:1, and 25:1, respectively. Correspondingly, the CTL activities by Ad mKDR were 65.2%+/-6.1%, 46.7%+/-5.0%, and 22.6%+/-3.7%. Sixty percent of the Ad hKDR-immunized mice with 5*10(6) Hepa 1-6 hepatoma cells were still alive two months after the inoculation, whereas just 40% of the Ad mKDR-immunized mice with 2*10(6) Hepa 1-6 cells survived two months. When CD8+ or CD4+ T lymphocytes were deleted in the mice the above mentioned CTL activities and protection of the mice from tumors disappeared. CONCLUSION: Adenovirus vector mediated xenogeneic KDR can effectively break the immune tolerance to hepatocellular carcinomas in an animal model and induce a strong antigen-specific T cell response, which is dependent on CD8+ and CD4+ T cells. PMID- 18423154 TI - [The application of 3D liver acquisition volume acceleration integrated with array spatial sensitivity encoding technique in liver dynamic-enhancement scanning]. AB - OBJECTIVE: To evaluate the application of 3D liver acquisition volume acceleration (3D-LAVA) integrated with array spatial sensitivity encoding technique (ASSET) in liver dynamic-enhancement scanning. METHODS: One hundred forty-seven patients underwent conventional plain and contrast enhancement liver MR imaging. 3D-LAVA and 2D fast spoiled gradient recalled echo were used for contrast enhancement liver MR scanning in 90 and 57 patients respectively. In the 3D-LAVA group, integrated ASSET was used in 72 out of the 90 patients. Of the 57 patients who underwent examinations using 2D fast spoiled gradient recalled echo, portal vein CE-MRA was performed on 20. The ability of 3D-LAVA to detect the lesions and the advantage to shorten the acquisition time after integrating with ASSET were analyzed. Original images of 60 patients in the 3D-LAVA group were processed using MIP to illustrate the anatomy of the portal vein. They were compared with those shown by CE-MRA to evaluate the illustration abilities of the two approaches. RESULTS: 3D-LAVA is more sensitive than 2D-FSPGR in detecting metastatic hepatic carcinomas. In the 3D-LAVA group integrated with ASSET, earlier and peak arterial phase images were acquired in 34 cases; and earlier, peak and late arterial phase images were acquired in 23 cases. The illustrations of the portal vein anatomy by 3D-LAVA were similar to those shown by portal vein CE-MRA. CONCLUSION: 3D-LAVA integrated with ASSET can obtain higher quality multi phase dynamic enhancement images of the liver in a shorter time, and in the meantime also shows the vascular anatomy. PMID- 18423155 TI - [Effect of ursolic acid on proliferation and apoptosis of hepatic stellate cells in vitro]. AB - OBJECTIVE: To investigate the effect of ursolic acid on proliferation and apoptosis of hepatic stellate cells (HSC) in vitro and explore the mechanisms of apoptosis of HSC induced by ursolic acid by studying the expressions of apoptosis regulating proteins Bcl-2, Bax and Caspase 3 in HSC. METHODS: Hepatic stellate cells HSC-T6 and hepatocytes L02 were incubated with different concentrations of ursolic acid (25, 50, 75, 100, 125 and 150 micromol/L) for 24 h, 48 h and 72 h. The effect of ursolic acid on the cell proliferation was studied by methyl thiazolyl tetrazolium (MTT) colorimetric assay. The rate of HSC-T6 apoptosis was identified by flow cytometry (FCM) and the morphological change of apoptosis was observed with light microscopy. The expressions of apoptosis-regulating protein Bcl-2, Bax and Caspase 3 in HSC-T6 after apoptosis induced by ursolic acid were examined by immunocytochemical staining assay. RESULTS: MTT analysis indicated administration of 25-150 micromol/L ursolic acid incubated with HSC-T6 for 24 h, 48 h and 72 h significantly inhibited HSC-T6 proliferation in a dose-dependent and time-dependent manner compared with the control group. Promotive effect of ursolic acid on proliferation of hepatocyte L02 was observed in the 25, 50, 75 micromol/L concentration groups. Ursolic acid inhibited L02 proliferation when its concentration was higher than 100 micromol/L and for 72 hours or longer. HE stained histological slides demonstrated morphologic changes of HSC-T6, including karyorrhexis and cytoplasm vacuolization, when they were treated with ursolic acid at 75 micromol/L concentrations for 48 h. FCM showed the apoptosis ratios of HSC-T6 were 10.30%+/-3.85%, 21.87%+/-4.46% and 31.33%+/-6.18% after treating HSC T6 with ursolic acid at concentrations of 25, 50 and 75 micromol/L for 48 h. They were significantly higher than that of the control group 2.93%+/-1.60%. Immunocytochemistry also indicated the expressions of Bax and caspase 3 protein in HSC-T6 cells were up-regulated in a dose-dependent manner, but expressions of Bcl-2 protein were not significantly different from that of the blank control group (P more than 0.05). CONCLUSIONS: Ursolic acid could significantly inhibit HSC proliferation and induce apoptosis in a dose-dependent and time-dependent manner. Ursolic acid in low concentration promotes proliferation of L02 cells, but in high concentrations (more than 100 micromol/L) it inhibits the growth of hepatocytes. Expressions of Bax and Caspase 3 in apoptotic HSC were increased; expressions of Bcl-2 protein were not significantly different from that of the control group, while Bcl-2/Bax ratio was reduced. Our results suggest that HSC-T6 cell apoptosis induced by ursolic acid occurs through mechanisms involving mitochondrial pathways and Bcl-2 family proteins. PMID- 18423156 TI - [Effect of zedoary turmeric oil on proliferation, apoptosis and extracellular matrix secretion of hepatic stellate cells]. PMID- 18423157 TI - [Screening of hepatocyte proteins binding to whole S protein of hepatitis B virus by yeast two-hybrid system]. PMID- 18423158 TI - [A preliminary study on side population cells of hepatocellular carcinomas]. PMID- 18423159 TI - [Peginterferon alpha-2a and ribavirin for treating chronic hepatitis C patients with persistent normal aminotransferase levels]. PMID- 18423160 TI - [An in vitro research on interleukin-1 beta promoted invasiveness of human hepatocellular carcinoma MHCC97-H cells]. PMID- 18423161 TI - [Relationship between the expression of uncoupling protein 2 and insulin resistance in fatty liver rats induced by high-fat food]. PMID- 18423163 TI - [Current status and perspectives of autologous liver transplantation]. PMID- 18423162 TI - [Clinical significance of the over-expression of tetraspanin CD151 in hepatocellular carcinomas]. PMID- 18423164 TI - [An experimental study on the expression and pathogenic effects of cyclooxygenase 2 in nonalcoholic fatty liver disease]. PMID- 18423165 TI - [Etiology of community-acquired lower urinary infections and antimicrobial resistance of Escherichia coli: a national surveillance study]. AB - BACKGROUND AND OBJECTIVES: The increasing rates of resistance exhibited by uropathogens represent a serious problem. The aim of this study was to determine, in Spain, the etiology of community-acquired lower urinary infections and antimicrobial resistance of Escherichia coli isolates. METHODS: Prospective multicenter study conducted between February and June 2006, in 15 microbiology laboratories located in 9 autonomous regions. RESULTS: A total of 3,109 isolates were recovered. E. coli was the most frequent (70.8%), followed by Klebsiella spp (6.8%) Proteus spp (6.6%), and Enterococcus spp (5.5%). The resistant rate of E. coli for phosphomycin was 1.7%, 3.8% nitrofurantoin, 6.9% cefexime, 8.1% amoxicillin-clavulanic, 8.9% cefuroxime, and 23.9% ciprofloxacin. The 5.2% were extended-spectrum beta-lactamase (ESBL)- producing microorganisms. Resistance of E. coli to ciprofloxacin was lower in people younger than 40 years (6.7% vs 33.9% in > 60, p < 0.001), and in some regions (12.5% in Galicia vs 37.3% in Valencia). ESBL-producing E.coli was higher in people older than 60 years (79.1% vs 7% in < 40, p < 0.001), and exhibited geographic variations (18.4% in Valencia, 0.8% in Galicia). The 68.6% of ESBL-producing E.coli were resistant to cotrimoxazole, 72.2% to ciprofloxacin vs 10.6% to nitrofurantoin and 1.9% to phosphomycin. CONCLUSIONS: The increasing rates of resistance and cross-resistance of this study make evident a real problem that strengthens the need for a reevaluation of the empiric treatment of lower urinary infections. PMID- 18423166 TI - [Validation of the Eating Attitudes Test (EAT) as a screening tool in the general population]. AB - BACKGROUND AND OBJECTIVE: To validate the best alternative cut-off point of a Spanish version of the Eating Attitudes Test (EAT-40) for the screening of eating disorders cases in the general population. SUBJECTS AND METHOD: The translated questionnaire was administered to a representative sample of 2,734 female students who ranged in age from 13 to 22 years. Participants who scored above 21 points were interviewed, as were a random sample of participants who scored 21 or below. Internal and external validity parameters were estimated for different alternative cut-off scores below the usual EAT score of 30. RESULTS: The best diagnostic prediction was obtained with a cut-off score of 21 points (sensitivity: 73.3%; specificity: 85.1%). A positive predictive value of 20% and a negative predictive value of 98.4% were obtained. CONCLUSIONS: The best diagnostic prediction in a non-clinical setting is obtained with a cut-off score of 21, below that recommended by the authors of the questionnaire. PMID- 18423167 TI - [Nasogastric enteral nutrition in severe acute pancreatitis]. AB - BACKGROUND AND OBJECTIVE: To establish the nasogastric enteral nutrition tolerance in patients with severe acute pancreatitis. PATIENTS AND METHOD: A total of 12 patients with severe acute pancreatitis (> or = 3 Ranson criteria; C reactive protein > 210 mg/dl) and adverse clinical course were included during 2006. When we verified the disease severity, nasogastric (10 F) enteral nutrition was initiated. We used a low fat semi-elemental feed (Dietgrif) in a slow infusion rate. We evaluated the enteral nutrition tolerance and the adverse events. RESULTS: The patient (4 women and 8 men) mean age (standard deviation) was 70 (11) years and the mean hospital stay was 86 days (range: 14-405 days). The etiology of pancreatitis was: gallstones 8, alcohol abuse one and unknown 3. All patients had medical and/or pancreatic complications. Seven had significant pancreatic necrosis detected in the abdominal computed tomography. Three patients were admitted in the critical care unit and 2 died. The nasogastric enteral nutrition was well tolerated in 8 out of 12 patients (67%) regardless of their medical or pancreatic complications. Only in 3 patients we had initially to discontinue the feeding because of ileus and total parenteral nutrition was provisionally necessary. Enteral nutrition was impossible in one patient because of duodenal stenosis. CONCLUSIONS: Nasogastric enteral nutrition is well tolerated in patients with severe acute pancreatitis and it is an alternative to others nutritional routes. PMID- 18423168 TI - [Community-acquired low urinary tract infection]. PMID- 18423169 TI - [Teledermatology]. PMID- 18423170 TI - [DNA arrays: a general overview and specific applications]. AB - Difficulties derived from the big complexity of the human genome (because of the great amount of genes) and difficulties at the time of performing a correct molecular diagnosis have led to the utilization of advanced technologies such as DNA arrays. By this technique, it is possible to analyze in a single experiment great sequences of genomic regions, to detect mutations or polymorphisms (characteristic of each individual), and to analyze variations in the expression of genes implicated in different disorders or in the utilization of drugs. Similarly, the use of DNA arrays-derived information has led to a considerable advance in the personalized medicine that is applied to every patient, avoiding unwanted adverse effects and the consecution of the best performance effectiveness for each drug. PMID- 18423171 TI - [Organization and functioning model of biomedical research structures. Situation and future challenges]. PMID- 18423173 TI - [Osteoporitic fracture risk factors in Spain]. PMID- 18423174 TI - [Human herpesvirus type 8 infection and development of cavity-based lymphoma in an old, HIV-negative patient with Kaposi's sarcoma]. PMID- 18423175 TI - [Subcutaneous emphysema and pneumomediastinum after dental extraction]. PMID- 18423176 TI - [Infectious mononucleosis]. PMID- 18423177 TI - [Diagnosis of respiratory diseases caused by asbestos]. PMID- 18423178 TI - [Association between chronic colonization or infection with Pseudomonas aeruginosa and bronchial hyperreactivity in patients with cystic fibrosis]. AB - OBJECTIVE: In patients with cystic fibrosis, bronchial hyperreactivity is a common finding that has not been conclusively associated with atopy. The objective of the present study was to determine the relationship between chronic colonization or infection with Pseudomonas aeruginosa and bronchial hyperreactivity in a group of patients with cystic fibrosis. PATIENTS AND METHODS: A nonspecific histamine bronchial provocation test was administered to a group of 32 cystic fibrosis patients with a mean (SD) age of 11.25 (3.7) years. The presence of atopy and of chronic colonization or infection with P aeruginosa was also studied. RESULTS: Nine of the 32 patients (28.1%) studied showed bronchial hyperreactivity. The clinical status of these 9 patients was significantly worse and all were colonized or infected with P aeruginosa. Atopy was present in 17 of the 32 patients (53.1%) in the study group, but in only 3 of the 9 patients (33.3%) with bronchial hyperreactivity. Bronchial hyperreactivity was significantly associated with colonization or infection with P aeruginosa (P< .001), but not with atopy (P=.12). In the patients without atopy, colonization was significantly associated with bronchial hyperreactivity (P=.017). In the group with normal lung function (forced expiratory volume in 1 second >/=80%) this association was also significant (P=.044), while the association between bronchial hyperreactivity and atopy was not (P=.11). CONCLUSIONS: The results of the present study suggest that in patients with cystic fibrosis, bronchial hyperreactivity may be associated with colonization or infection with P aeruginosa, and that this may be a more important risk factor for bronchial hyperreactivity than atopy. PMID- 18423179 TI - [Diagnoses and diagnostic procedures in 500 consecutive patients with clinical suspicion of interstitial lung disease]. AB - OBJECTIVE: To determine the diagnostic yield achieved with the application of current recommendations for evaluating patients with suspected interstitial lung disease (ILD) and the procedures that must be applied to reach a definitive diagnosis. PATIENTS AND METHODS: Over a 10-year period, 500 consecutive patients attending an ILD outpatient clinic who showed features of diffuse lung involvement were assessed with a single diagnostic protocol. Results were introduced in a dedicated database and diagnoses for idiopathic interstitial pneumonia were established according to a recent consensus classification. RESULTS: A definitive diagnosis was reached in 427 (85%) patients: in 125 without invasive procedures and in 302 with invasive procedures. In 73 (14.6%) cases a definitive diagnosis was not reached, and patients were placed in the group of unclassifiable interstitial pneumonia. Idiopathic interstitial pneumonia was the predominant group with 193 (39%) patients. The main specific entities included sarcoidosis with 93 (19%) patients, usual interstitial pneumonia with 84 (17%) patients, and hypersensitivity pneumonitis with 75 (15%) patients. Thirty (6%) patients were diagnosed with an illness other than ILD (false ILD). In 332 patients, we performed a total of 433 invasive procedures: transbronchial biopsy in 252 (direct diagnostic yield, 38%, or if used also to exclude other specific diagnosis, 50%), bronchoalveolar lavage in 260 (yield, 5%), and open lung biopsy in 141 (yield, 93%). Hence, following the current diagnostic approach, a definitive diagnosis was established for 85% of patients, for 25% solely on clinical grounds and imaging criteria and for 60% on the basis of invasive procedures. Diagnosis by open lung biopsy was still required for 141 (28%) patients. CONCLUSIONS: The diagnostic yield was high when the recommended study protocol was followed. A quarter of the diagnoses were reached with clinical criteria alone, but another quarter could only be made after open lung biopsy. PMID- 18423180 TI - [Management according to the Global Initiative for Asthma guidelines of patients with near-fatal asthma reduces morbidity and mortality]. AB - OBJECTIVE: To assess the effect of adequate outpatient care as defined by guidelines of the Global Initiative for Asthma (GINA) on the long-term outcome of near-fatal asthma. PATIENTS AND METHODS: Fifty-three patients who had experienced a near-fatal attack of asthma were treated according to the GINA guidelines and followed for a mean of 49 months (intervention group). Clinical and spirometric measurements corresponding to the periods before the attack (obtained retrospectively) and after the attack (obtained prospectively) were compared to measurements from 40 near-fatal asthma patients who had not been managed according to the GINA guidelines and who were followed for a mean of 51 months (historic control group). RESULTS: There were no deaths in the intervention group and 6 deaths (15%) in the control group (P=.005). The mean (SD) number of new near-fatal asthma attacks was significantly lower in the intervention group (0.17 [0.61]) than in the control group (1.6 [1]) (P< .001). Emergency visits following a near-fatal asthma attack decreased from 0.9 (1.8) to 0.3 (0.6) in the intervention group and hospital admissions decreased from 3.4 (5.1) to 0.5 (1.4) (P< .001). Eosinophil count decreased from 390 (411) x l0(9) cells/L to 159 (121) x l0(9) cells/L (P=.01) and forced expiratory volume in 1 second increased from 68% (23%) of predicted to 76% (20%) (P=.006). CONCLUSIONS: Management according to the GINA guidelines of patients who had experienced a near-fatal asthma attack was associated with a decrease in asthma morbidity and mortality. PMID- 18423181 TI - [Traumatic rupture of the diaphragm]. AB - OBJECTIVE: Traumatic rupture of the diaphragm (TRD) is a rare occurrence, with variable morbidity and mortality. The aim of this study was to analyze cases of TRD in a tertiary hospital and assess prognostic factors associated with mortality. PATIENTS AND METHODS: A retrospective study was performed of patients diagnosed with TRD in Hospital Universitario La Fe, Valencia, Spain, between 1969 and 2006. The following variables were analyzed: sex, age, cause, diagnosis, associated lesions, surgical procedure, side and size of the lesion, visceral herniation, and postoperative morbidity and mortality. RESULTS: The study group comprised 132 patients (105 men, 79.5%) with a mean (SD) age of 39.64 (17.04) years. Traffic accidents were the most common cause of TRD. Rupture involved the left hemidiaphragm in 96 cases (72.7%), and 113 patients (85.6%) had associated lesions, most often affecting the abdomen. Thoracotomy was performed in 83 cases (62.9%) and laparotomy in 41 (31.1%). Visceral herniation was reported in 90 patients (68.3%), most often involving the stomach. The rates of perioperative morbidity and mortality were 62.8% and 20.5%, respectively. Diagnostic delay and the presence of morbidity and serious associated lesions all had a statistically significant impact on mortality (P< .05). In the case of serious associated lesions, the odds ratio was 2.898 (95% confidence interval, 1.018-8.250) and for perioperative morbidity it was 1.488 (95% confidence interval, 1.231-1.798). CONCLUSIONS: TRD is an infrequent occurrence in young men, is generally caused by traffic accidents, and is more common on the left side. Associated lesions are present in most cases and represent the main prognostic factor affecting morbidity and mortality. TRD can be considered a relative surgical emergency when not accompanied by other lesions that in themselves constitute surgical emergencies. PMID- 18423182 TI - [The role of the pulmonologist in the assessment of disability in patients with respiratory disease]. AB - A high percentage of the individuals who consult respiratory medicine specialists are working adults, and these patients often complain that dyspnea on exertion hinders their ability to do their job. In other cases, patients are referred for assessment by those responsible for evaluating disability. Furthermore, diagnosis of a respiratory disease should be accompanied by a therapeutic regimen which, in addition to pharmacotherapy, should include advice on lifestyle, nutrition, and physical exercise and recommendations on the kind of work the patient can undertake. Pulmonologists are therefore heavily involved in the assessment of disability in patients with respiratory diseases. The aim of this review is to offer respiratory specialists a broad view of the medical and legal procedures used to evaluate functional impairment caused by respiratory disease, and suggest how they can make recommendations to these patients concerning the appropriate working conditions for the prevention and treatment of their disease. PMID- 18423183 TI - [Recommendations for pharmacological tobacco cessation treatments: proposals for financing]. PMID- 18423184 TI - [Resection of 8 mediastinal bronchogenic cysts by video-assisted thoracoscopy]. AB - Mediastinal bronchogenic cysts are an uncommon entity and surgical experience of their removal by video-assisted thoracoscopy is limited. We present our patient outcomes and surgical technique in the treatment of bronchogenic cysts by video assisted thoracoscopy. The study included 8 patients (4 females and 4 males between the ages of 4 and 52 years), 7 of whom presented clinical symptoms. The mean widest diameter of the cyst was 7.6 cm. In 5 patients the cyst was in the middle mediastinum and in 3, the posterior mediastinum. The intervention was performed using 3 or 4 entry points. Initial puncture of the cyst and removal of its contents greatly facilitated cyst manipulation and subsequent dissection of the cyst sac from the structures to which it was attached. In all 8 cases resection by video-assisted thoracoscopy was carried out with no intraoperative complications. The mean postoperative hospital stay was 3.3 days. During follow up, which ranged from 4 months to 10 years, no patients presented late-onset or recurrent complications. PMID- 18423185 TI - [Value of thoracoscopy in the diagnosis and treatment of complicated thoracic endometriosis in 2 patients]. AB - We present 2 cases in which, when faced with suspicion of catamenial hemopneumothorax or pneumothorax, video-assisted thoracoscopy was used for identification, biopsy, and electrocoagulation of the tissue sites where pleural endometriosis was suspected. PMID- 18423186 TI - [Hypersensitivity pneumonitis associated with Mycobacterium chelonae]. PMID- 18423187 TI - [Cystic adenomatoid malformation of the lungs in adults]. PMID- 18423188 TI - India takes an open source approach to drug discovery. AB - Open source software may have been around for 17 years, but using an open source model to speed up drug discovery is a relatively new idea. This month, India is launching a new open source initiative for developing drugs to treat diseases such as tuberculosis, malaria, and HIV. PMID- 18423189 TI - Rethinking pseudokinases. AB - Pseudokinases lack conservation of one or more of the catalytic residues in the kinase core and as a consequence are typically thought to be catalytically inactive. New work by Mukherjee et al. (2008) challenges this assumption. They show that the pseudokinase domain of CASK (Ca2+/calmodulin activated serine threonine kinase) adopts an active conformation and displays catalytic activity in vivo. PMID- 18423190 TI - O2 sensing: only skin deep? AB - The transcription factor HIF-1 mediates adaptive responses to hypoxia, and its activity is negatively regulated by O2-dependent binding of the von Hippel-Lindau (VHL) protein. In this issue, Boutin et al. (2008) use conditional knockout mice to demonstrate that sensing of O2 by keratinocytes in the epidermis leads to alterations in cutaneous blood flow that affect the production of the hormone erythropoietin, thereby modulating red blood cell production and the O2-carrying capacity of blood. PMID- 18423191 TI - A TRIFfic perspective on acute lung injury. AB - Acute lung injury (ALI) is a leading cause of death in people infected with H5N1 avian influenza virus or the SARS-coronavirus. Imai et al. (2008) now report that ALI is triggered by the signaling of oxidized phospholipids through Toll-like receptor 4 (TLR4) and the adaptor protein TRIF. These findings provide insight into the molecular pathogenesis of ALI, a condition for which treatment options are currently very limited. PMID- 18423192 TI - Fly courtship song: triggering the light fantastic. AB - In a study in this issue, Clyne and Miesenbock (2008) apply an ingenious optogenetic technology to activate neurons that generate male-specific courtship song in flies. This work sheds new light on the neural circuitry underlying sexually dimorphic behaviors in Drosophila. PMID- 18423193 TI - EJCs at the heart of translational control. AB - In mammalian cells, the splicing machinery deposits the exon junction complex (EJC) on mRNA splice junctions. Two studies in this issue now link the EJC to different aspects of translational control. Ma et al. (2008) show that the EJC activates translation downstream of the mTOR signaling pathway, whereas Isken et al. (2008) establish that translation is repressed by partners of the EJC that are implicated in nonsense mediated decay (NMD). PMID- 18423194 TI - miRiad roles for the miR-17-92 cluster in development and disease. AB - MicroRNAs (miRNAs) encoded by the miR-17-92 cluster and its paralogs are known to act as oncogenes. Expression of these miRNAs promotes cell proliferation, suppresses apoptosis of cancer cells, and induces tumor angiogenesis. New work reveals essential functions for these miRNAs not only in tumor formation but also during normal development of the heart, lungs, and immune system. PMID- 18423195 TI - Epidermal sensing of oxygen is essential for systemic hypoxic response. AB - Skin plays an essential role, mediated in part by its remarkable vascular plasticity, in adaptation to environmental stimuli. Certain vertebrates, such as amphibians, respond to hypoxia in part through the skin; but it is unknown whether this tissue can influence mammalian systemic adaptation to low oxygen levels. We have found that epidermal deletion of the hypoxia-responsive transcription factor HIF-1alpha inhibits renal erythropoietin (EPO) synthesis in response to hypoxia. Conversely, mice with an epidermal deletion of the von Hippel-Lindau (VHL) factor, a negative regulator of HIF, have increased EPO synthesis and polycythemia. We show that nitric oxide release induced by the HIF pathway acts on cutaneous vascular flow to increase systemic erythropoietin expression. These results demonstrate that in mice the skin is a critical mediator of systemic responses to environmental oxygen. PMID- 18423196 TI - Identification of oxidative stress and Toll-like receptor 4 signaling as a key pathway of acute lung injury. AB - Multiple lung pathogens such as chemical agents, H5N1 avian flu, or SARS cause high lethality due to acute respiratory distress syndrome. Here we report that Toll-like receptor 4 (TLR4) mutant mice display natural resistance to acid induced acute lung injury (ALI). We show that TLR4-TRIF-TRAF6 signaling is a key disease pathway that controls the severity of ALI. The oxidized phospholipid (OxPL) OxPAPC was identified to induce lung injury and cytokine production by lung macrophages via TLR4-TRIF. We observed OxPL production in the lungs of humans and animals infected with SARS, Anthrax, or H5N1. Pulmonary challenge with an inactivated H5N1 avian influenza virus rapidly induces ALI and OxPL formation in mice. Loss of TLR4 or TRIF expression protects mice from H5N1-induced ALI. Moreover, deletion of ncf1, which controls ROS production, improves the severity of H5N1-mediated ALI. Our data identify oxidative stress and innate immunity as key lung injury pathways that control the severity of ALI. PMID- 18423197 TI - Direct reprogramming of terminally differentiated mature B lymphocytes to pluripotency. AB - Pluripotent cells can be derived from fibroblasts by ectopic expression of defined transcription factors. A fundamental unresolved question is whether terminally differentiated cells can be reprogrammed to pluripotency. We utilized transgenic and inducible expression of four transcription factors (Oct4, Sox2, Klf4, and c-Myc) to reprogram mouse B lymphocytes. These factors were sufficient to convert nonterminally differentiated B cells to a pluripotent state. However, reprogramming of mature B cells required additional interruption with the transcriptional state maintaining B cell identity by either ectopic expression of the myeloid transcription factor CCAAT/enhancer-binding-protein-alpha (C/EBPalpha) or specific knockdown of the B cell transcription factor Pax5. Multiple iPS lines were clonally derived from both nonfully and fully differentiated B lymphocytes, which gave rise to adult chimeras with germline contribution, and to late-term embryos when injected into tetraploid blastocysts. Our study provides definite proof for the direct nuclear reprogramming of terminally differentiated adult cells to pluripotency. PMID- 18423198 TI - The 3D structure of the immunoglobulin heavy-chain locus: implications for long range genomic interactions. AB - The immunoglobulin heavy-chain (Igh) locus is organized into distinct regions that contain multiple variable (V(H)), diversity (D(H)), joining (J(H)) and constant (C(H)) coding elements. How the Igh locus is structured in 3D space is unknown. To probe the topography of the Igh locus, spatial distance distributions were determined between 12 genomic markers that span the entire Igh locus. Comparison of the distance distributions to computer simulations of alternative chromatin arrangements predicted that the Igh locus is organized into compartments containing clusters of loops separated by linkers. Trilateration and triple-point angle measurements indicated the mean relative 3D positions of the V(H), D(H), J(H), and C(H) elements, showed compartmentalization and striking conformational changes involving V(H) and D(H)-J(H) elements during early B cell development. In pro-B cells, the entire repertoire of V(H) regions (2 Mbp) appeared to have merged and juxtaposed to the D(H) elements, mechanistically permitting long-range genomic interactions to occur with relatively high frequency. PMID- 18423199 TI - Meiosis I is established through division-specific translational control of a cyclin. AB - In budding yeast, key meiotic events such as DNA replication, recombination, and the meiotic divisions are controlled by Clb cyclin-dependent kinases (Clb-CDKs). Using a novel synchronization procedure, we have characterized the activity of these Clb-CDKs and observed a surprising diversity in their regulation during the meiotic divisions. Clb1-CDK activity is restricted to meiosis I, and Clb3-CDK activity to meiosis II, through 5'UTR-mediated translational control of its transcript. The analysis of cells inappropriately producing Clb3-CDKs during meiosis I furthermore defines Clb3 as an inhibitor of the meiosis I chromosome segregation program. Our results demonstrate an essential role for Clb-CDK regulation in establishing the meiotic chromosome segregation pattern. PMID- 18423200 TI - Yeast life span extension by depletion of 60s ribosomal subunits is mediated by Gcn4. AB - In nearly every organism studied, reduced caloric intake extends life span. In yeast, span extension from dietary restriction is thought to be mediated by the highly conserved, nutrient-responsive target of rapamycin (TOR), protein kinase A (PKA), and Sch9 kinases. These kinases coordinately regulate various cellular processes including stress responses, protein turnover, cell growth, and ribosome biogenesis. Here we show that a specific reduction of 60S ribosomal subunit levels slows aging in yeast. Deletion of genes encoding 60S subunit proteins or processing factors or treatment with a small molecule, which all inhibit 60S subunit biogenesis, are each sufficient to significantly increase replicative life span. One mechanism by which reduced 60S subunit levels leads to life span extension is through induction of Gcn4, a nutrient-responsive transcription factor. Genetic epistasis analyses suggest that dietary restriction, reduced 60S subunit abundance, and Gcn4 activation extend yeast life span by similar mechanisms. PMID- 18423201 TI - SKAR links pre-mRNA splicing to mTOR/S6K1-mediated enhanced translation efficiency of spliced mRNAs. AB - Different protein complexes form on newly spliced mRNA to ensure the accuracy and efficiency of eukaryotic gene expression. For example, the exon junction complex (EJC) plays an important role in mRNA surveillance. The EJC also influences the first, or pioneer round of protein synthesis through a mechanism that is poorly understood. We show that the nutrient-, stress-, and energy-sensing checkpoint kinase, mTOR, contributes to the observed enhanced translation efficiency of spliced over nonspliced mRNAs. We demonstrate that, when activated, S6K1 is recruited to the newly synthesized mRNA by SKAR, which is deposited at the EJC during splicing, and that SKAR and S6K1 increase the translation efficiency of spliced mRNA. Thus, SKAR-mediated recruitment of activated S6K1 to newly processed mRNPs serves as a conduit between mTOR checkpoint signaling and the pioneer round of translation when cells exist in conditions supportive of protein synthesis. PMID- 18423202 TI - Upf1 phosphorylation triggers translational repression during nonsense-mediated mRNA decay. AB - In mammalian cells, nonsense-mediated mRNA decay (NMD) generally requires that translation terminates sufficiently upstream of a post-splicing exon junction complex (EJC) during a pioneer round of translation. The subsequent binding of Upf1 to the EJC triggers Upf1 phosphorylation. We provide evidence that phospho Upf1 functions after nonsense codon recognition during steps that involve the translation initiation factor eIF3 and mRNA decay factors. Phospho-Upf1 interacts directly with eIF3 and inhibits the eIF3-dependent conversion of 40S/Met tRNA(i)(Met)/mRNA to translationally competent 80S/Met-tRNA(i)(Met)/mRNA initiation complexes to repress continued translation initiation. Consistent with phospho-Upf1 impairing eIF3 function, NMD fails to detectably target nonsense containing transcripts that initiate translation independently of eIF3 from the CrPV IRES. There is growing evidence that translational repression is a key transition that precedes mRNA delivery to the degradation machinery. Our results uncover a critical step during NMD that converts a pioneer translation initiation complex to a translationally compromised mRNP. PMID- 18423203 TI - CASK Functions as a Mg2+-independent neurexin kinase. AB - CASK is a unique MAGUK protein that contains an N-terminal CaM-kinase domain besides the typical MAGUK domains. The CASK CaM-kinase domain is presumed to be a catalytically inactive pseudokinase because it lacks the canonical DFG motif required for Mg2+ binding that is thought to be indispensable for kinase activity. Here we show, however, that CASK functions as an active protein kinase even without Mg2+ binding. High-resolution crystal structures reveal that the CASK CaM-kinase domain adopts a constitutively active conformation that binds ATP and catalyzes phosphotransfer without Mg2+. The CASK CaM-kinase domain phosphorylates itself and at least one physiological interactor, the synaptic protein neurexin-1, to which CASK is recruited via its PDZ domain. Thus, our data indicate that CASK combines the scaffolding activity of MAGUKs with an unusual kinase activity that phosphorylates substrates recuited by the scaffolding activity. Moreover, our study suggests that other pseudokinases (10% of the kinome) could also be catalytically active. PMID- 18423204 TI - Rac1 activation controls nuclear localization of beta-catenin during canonical Wnt signaling. AB - Canonical Wnt signaling critically regulates cell fate and proliferation in development and disease. Nuclear localization of beta-catenin is indispensable for canonical Wnt signaling; however, the mechanisms governing beta-catenin nuclear localization are not well understood. Here we demonstrate that nuclear accumulation of beta-catenin in response to Wnt requires Rac1 activation. The role of Rac1 depends on phosphorylation of beta-catenin at Ser191 and Ser605, which is mediated by JNK2 kinase. Mutations of these residues significantly affect Wnt-induced beta-catenin nuclear accumulation. Genetic ablation of Rac1 in the mouse embryonic limb bud ectoderm disrupts canonical Wnt signaling and phenocopies deletion of beta-catenin in causing severe truncations of the limb. Finally, Rac1 interacts genetically with beta-catenin and Dkk1 in controlling limb outgrowth. Together these results uncover Rac1 activation and subsequent beta-catenin phosphorylation as a hitherto uncharacterized mechanism controlling canonical Wnt signaling and may provide additional targets for therapeutic intervention of this important pathway. PMID- 18423205 TI - Sex-specific control and tuning of the pattern generator for courtship song in Drosophila. AB - The differentially spliced transcription factors encoded by the fruitless (fru) gene are key determinants of sexual behavior in Drosophila. They are expressed in a minority of neurons with limited dimorphisms and regulate neural processes that remain largely unknown. Here, we use light-activated ion channels to stimulate fru-expressing neurons in the thoracic-abdominal ganglia, enabling direct functional comparisons of homologous circuitry between sexes. Optical stimulation of males or females initiates the unilateral wing vibrations that normally generate the male courtship song. The pattern-generating circuit operates differently in the two sexes, producing wing movement and sound in both but authentic songs only in males and in females expressing male fru product. A song like motor program is thus present in females but lies dormant because the neural commands required for song initiation are absent. Supplying such commands artificially reveals fru-specific differences in the internal dynamics of the song generator and sets the stage for exploring their physiological basis. PMID- 18423206 TI - A quantitative spatiotemporal atlas of gene expression in the Drosophila blastoderm. AB - To fully understand animal transcription networks, it is essential to accurately measure the spatial and temporal expression patterns of transcription factors and their targets. We describe a registration technique that takes image-based data from hundreds of Drosophila blastoderm embryos, each costained for a reference gene and one of a set of genes of interest, and builds a model VirtualEmbryo. This model captures in a common framework the average expression patterns for many genes in spite of significant variation in morphology and expression between individual embryos. We establish the method's accuracy by showing that relationships between a pair of genes' expression inferred from the model are nearly identical to those measured in embryos costained for the pair. We present a VirtualEmbryo containing data for 95 genes at six time cohorts. We show that known gene-regulatory interactions can be automatically recovered from this data set and predict hundreds of new interactions. PMID- 18423207 TI - SnapShot: The TGFbeta pathway interactome. PMID- 18423208 TI - Mass spectrometric characterization of proteins modified by nitric oxide-derived species. AB - Nitric oxide-derived metabolites have been demonstrated to covalently modify cellular protein repertoire, thus affecting specific enzymatic functions. Among the various redox posttranslational modifications, protein nitration has been broadly recognized by immunological, spectroscopical, and chromatographic methods as a widespread reaction regulating essential phatophysiological processes. With the introduction of matrix-assisted laser desorption and electrospray as soft ionization methods for mass spectrometry of biomolecules, nitration has been investigated directly at the protein level, assigning polypeptide modification sites. Peptide mass fingerprinting and fragment fingerprinting upon collisional fragmentation approaches have been widely used to this purpose. This chapter describes how minimal levels of nitration present on a model protein, namely bovine serum albumin, generated in vitro by ONOO(-) treatment, were ascertained by integrated mass spectrometry approaches, identifying sites of modification. Critical considerations on the limits of each mass spectrometric ionization methods are also provided. PMID- 18423209 TI - Detecting nitrated proteins by proteomic technologies. AB - Nitration is a posttranslational modification of tyrosine residues of proteins mediated by peroxynitrite (ONOO(-)). It commonly occurs in neurological and pathological disorders, which involve nitric oxide (NO)-mediated oxidative stress. Nitration of tyrosine or tyrosyl groups of a protein modulates protein function and initiates signal transduction pathways, which lead to alternation of cellular metabolism and functions. Because of its apparent significance, there is an increasing urge to identify nitrated proteins as a bridge to expand our understanding of their involvement in different biological processes. This chapter describes strategies that could be used for rapid screening and detection of nitrated proteins, subsequent resolution, and identification of nitrated proteins and peptides using proteomic technologies. These include two-dimensional gel electrophoresis coupled with Western blotting and matrix-assisted laser desorption/ionization time of flight mass spectrometry, as well as liquid chromatography-linked tandem mass spectrometry. PMID- 18423210 TI - Using tandem mass spectrometry to quantify site-specific chlorination and nitration of proteins: model system studies with high-density lipoprotein oxidized by myeloperoxidase. AB - Protein oxidation is implicated in atherogenesis and other inflammatory conditions. Measuring levels of chlorinated and nitrated proteins in biological matrices serves as a quantitative index of oxidative stress in vivo. One potential mechanism for oxidative stress involves myeloperoxidase, a heme protein expressed by neutrophils, monocytes, and some populations of macrophages. The enzyme uses hydrogen peroxide to generate an array of cytotoxic oxidants, including hypochlorous acid (HOCl), a potent chlorinating intermediate, and nitrogen dioxide radical, a reactive nitrogen species (RNS). One important target may be high-density lipoprotein (HDL), which is implicated in atherogenesis. This chapter describes liquid chromatography-tandem mass spectrometric methods for quantifying site-specific modifications of proteins that have been oxidized by HOCl or RNS. Our studies center on apolipoprotein A-I, the major HDL protein, which provides an excellent model system for investigating factors that target specific residues for oxidative damage. Our approach is sensitive and rapid, applicable to a wide array of posttranslational modifications, and does not require peptides to be derivatized or labeled with an isotope. PMID- 18423211 TI - Influence of intramolecular electron transfer mechanism in biological nitration, nitrosation, and oxidation of redox-sensitive amino acids. AB - Using both high-performance liquid chromatography (HPLC) and electron spin resonance (ESR) spin-trappng techniques, we developed an analytical methodology for investigating intramolecular electron transfer-mediated tyrosyl nitration and cysteine nitrosation in model peptides. Peptides N-acetyl-TyrCys-amide (YC), N acetyl-TyrAlaCys-amide, N-acetyl-TyrAlaAlaCys-amide, and N-acetyl TyrAlaAlaAlaAlaCys-amide were used as models. Product analysis showed that nitration and oxidation products derived from YC and related peptides in the presence of myeloperoxidase (MPO)/H(2)O(2)/NO(2)(-) were not detectable. The major product was determined to be the corresponding disulfide (e.g., YCysCysY), suggestive of a rapid electron transfer from the tyrosyl radical to the cysteinyl residue. ESR spin-trapping experiments with 5,5'-dimethyl-1-pyrroline N-oxide (DMPO) demonstrated that thiyl radical intermediates were formed from peptides (e.g., YC) treated with MPO/H(2)O(2) and MPO/H(2)O(2)/NO(2)(-). Blocking the thiol group in YC totally abrogated thiyl radical formation. Under similar conditions, we were, however, able to trap the tyrosyl radical using the spin trap dibromonitrosobenzene sulfonic acid (DBNBS). Competition spin-trapping experiments revealed that intramolecular electron transfer is the dominant mechanism for thiyl radical formation in YC peptides. We conclude that a rapid intramolecular electron transfer mechanism between redox-sensitive amino acids could influence both protein nitration and nitrosation reactions. This mechanism brings together nitrative, nitrosative, and oxidative mechanisms in free radical biology. PMID- 18423212 TI - Protein thiol modification by peroxynitrite anion and nitric oxide donors. AB - Oxidation and modification of protein cysteines can have profound effects on protein structure and function. Using tubulin and microtubule-associated proteins (MAP) tau and MAP2 as examples, this chapter summarizes methods employed to characterize total cysteine modification using thiol-specific reagent 5 iodoacetamido-fluorescein labeling. Western blot analysis of peroxynitrite damaged tubulin under nonreducing conditions reveals the formation of higher molecular weight dimers and tetramers. Disulfides in microtubule proteins are substrates for both the thioredoxin reductase system and the glutaredoxin/glutathione reductase system. The yield of disulfides formed by peroxynitrite anion is quantitated by monitoring the oxidation of NADPH, a cofactor required by the thioredoxin reductase system. Treatment of proteins with S-nitrosothiols, including S-nitrosoglutathione and S-nitroso-N-acetyl penicillamine, can yield either disulfides or protein S-nitrosation. In the case of tubulin, both types of cysteine modification were detected. PMID- 18423213 TI - Indirect mechanisms of DNA strand scission by peroxynitrite. AB - Peroxynitrite is a highly reactive nitrogen species directly damaging diverse biomolecules in target cells. This is not the case for genomic DNA, as its cleavage is mediated by secondary reactive species produced at the mitochondrial level in a Ca(2+)-dependent reaction in which ubisemiquinone serves as an electron donor. Under these conditions, superoxide is produced in a time dependent manner for up to 30 min and dismutates readily to H(2)O(2), which can now reach the nucleus and generate DNA strand scission in a reaction of the Fenton type. This chapter provides evidence for this mechanism using the alkaline halo assay, a simple technique for the assessment of DNA strand scission at the single cell level. PMID- 18423214 TI - Nitric oxide: interaction with the ammonia monooxygenase and regulation of metabolic activities in ammonia oxidizers. AB - The biological nitrogen cycle is a complex interplay of many microorganisms. In the past, oxidation of the inorganic nitrogen compound ammonia by the ammonia oxidizing bacteria was thought to be restricted to oxic environments, and the metabolic flexibility of these organisms seemed to be limited. The discovery of an anaerobic metabolism in the late 1990s showed that these assumptions are no longer valid. NO and NO(2) are essential intermediates in ammonia oxidation. Both gases have wide-ranging regulatory effects on ammonia oxidation, denitrification, and biofilm formation of the ammonia oxidizing bacteria. PMID- 18423215 TI - Chemiluminescent detection of S-nitrosated proteins: comparison of tri-iodide, copper/CO/cysteine, and modified copper/cysteine methods. AB - The precise quantification of high and low molecular weight S-nitrosothiols (RSNOs) in biological samples is necessary for the study of nitric oxide dependent posttranslational signal transduction. Several chemiluminescence-based methods are used for the detection of S-nitrosothiols using the nitric oxide analyzer, including the tri-iodide method and the Cu/CO/cysteine (3C) method. Despite the fact that the tri-idodide method is the most widely used and validated methodology, the levels of S-nitrosated hemoglobin (SNO-Hb) and S nitrosated albumin have been lower than those reported using photolysis coupled to chemiluminescence. This chapter demonstrates that the tri-iodide method and a newly developed modified copper/cysteine (2C) method compare favorably with the 3C method. Our comparisons include physiologically relevant conditions where the ratio of SNO to heme is low and the frequency of nitrosation of a given Hb tetramer is less than 1. In our studies, the tri-iodide method, the 3C method, and the modified 2C method give consistent and reproducible results. These studies suggest that the proper use of any of these methods can be effective in the accurate measurement of S-nitrosothiols in biological samples. Using more than one in combination has the potential to resolve controversies related to the role of hemoglobin in the generation of RSNOs or the role of RSNOs in biology, whether the RSNOs derive from nitrite or other nitrosative pathways. PMID- 18423216 TI - S-nitrosothiol assays that avoid the use of iodine. AB - S-Nitrosylation is a ubiquitous signaling process in biological systems. Research regarding this signaling has been hampered, however, by assays that lack sensitivity and specificity. In particular, iodine-based assays for S nitrosothiols (1) produce nitrosyliodide, a potent nitrosating agent that can be lost to reactions in the biological sample being studied; (2) require pretreatment of biological samples with several reagents that react with proteins, artifactually forming or breaking S-NO bonds before the assay; and (3) are not sensitive or specific for nitrogen oxides in biological samples, reporting a wide range of different concentrations and falsely reporting NO modified proteins, to be nitrite. These data, therefore, suggest that iodine based assays should never be used for biological S-nitrosothiols. There are other assays that provide reasonably sensitive and accurate data regarding biological S nitrosothiols, including assays based on mass spectrometry, spectrophotometry, chemiluminescence, fluorescence, and immunostaining. Each assay, however, has limitations and should be quantitatively complemented by separate assays. Continued improvement in assays will facilitate improved understanding of S nitrosylation signaling. PMID- 18423217 TI - Analysis of citrulline, arginine, and methylarginines using high-performance liquid chromatography. AB - Citrulline is a product of arginine degradation by nitric oxide synthase and is a precursor for arginine synthesis in animal cells. After arginine is incorporated into proteins, it may undergo methylation to form N(G)-monomethylarginine, which may be converted to asymmetric dimethylarginine and symmetric dimethylarginine. The degradation of these methylated proteins produces free methylarginines. This chapter focuses on the analysis of these amino acids in biological samples (including plasma/serum, urine, cell culture medium, and tissues) using high performance liquid chromatography that involves precolumn derivatization with o phthaldialdehyde. Fluorescence is monitored at excitation and emission wavelengths of 340 and 455 nm, respectively. Detection limits are 5 nM for amino acids. The assays are linear between 1 and 100 microM for citrulline and arginine and between 0.1 and 10 microM for methylarginines. These chromatographic methods are highly sensitive, specific, accurate, and easily automated and provide a useful tool to study the regulation of the arginine-nitric oxide pathway. PMID- 18423218 TI - Quantitative proteome mapping of nitrotyrosines. AB - An essential first step in the understanding disease and environmental perturbations is the early and quantitative detection of the increased levels of the inflammatory marker nitrotyrosine, as compared with its endogenous levels within the tissue or cellular proteome. Thus, methods that successfully address a proteome-wide quantitation of nitrotyrosine and related oxidative modifications can provide early biomarkers of risk and progression of disease, as well as effective strategies for therapy. Multidimensional separations LC coupled with tandem mass spectrometry (LC-MS/MS) has, in recent years, significantly expanded our knowledge of human (and mammalian model system) proteomes, including some nascent work in identification of posttranslational modifications. This chapter discusses the application of LC-MS/MS for quantitation and identification of nitrotyrosine-modified proteins within the context of complex protein mixtures presented in mammalian proteomes. PMID- 18423219 TI - Protein S-nitrosation in signal transduction: assays for specific qualitative and quantitative analysis. AB - S-Nitrosation is a type of protein posttranslational modification that has now been found comparable with phosphorylation and acetylation in terms of its contribution to the intracellular signaling networks associated with pathological cell reactions such as host-pathogen interactions, low oxygen availability, cell cycle arrest, and programmed cell death. Therefore, elegant approaches are required to analyze endogenous S-nitrosation of intracellular proteins employed in the regulation of intracellular signal transduction. This chapter describes and discusses recently developed methods that allow both qualitative and quantitative analyses of S-nitrosation of intracellular proteins based on Western blot and enzyme-linked immunosorbent assay. PMID- 18423220 TI - Determination of mammalian arginase activity. AB - Of all arginine catabolic enzymes, the arginases and nitric oxide (NO) synthases are the ones that are of greatest interest to many investigators. Mammalian arginases catalyze the hydrolysis of arginine to ornithine and urea and are composed of two distinct isozymes: arginase I, located within the cytosol, and arginase II, located within mitochondria. The arginases not only can inhibit NO synthesis by reducing arginine availability, but also can promote the synthesis of polyamines or proline via production of the common precursor ornithine. Because of their inducibility in many cell types and to their potential impact on multiple biochemical pathways in health and disease, there is growing interest in assays of arginase activity. Although arginase activity may be determined by either spectrophotometric or radiochemical assays, radiochemical assays afford greater sensitivity and do not require correction for any ornithine or urea that may be present in the samples. Part of the arginase assay protocol described in this chapter also can be used for radiochemical assays of enzymes that catalyze decarboxylation reactions. No activity assay currently available is capable of distinguishing the arginase isozymes. PMID- 18423221 TI - Measurement of protein S-nitrosylation during cell signaling. AB - S-Nitrosylation, the modification of a cysteine thiol by a nitric oxide (NO) group, has emerged as an important posttranslational modification of signaling proteins. An impediment to studying the regulation of cell signaling by S nitrosylation has been the technical challenge of detecting endogenously S nitrosylated proteins. Detection of S-nitrosylated proteins is difficult because the S-NO bond is labile and therefore can be lost or gained artifactually during sample preparation. Nevertheless, several methods have been developed to measure endogenous protein S-nitrosylation, including the biotin switch assay and the chemical reduction/chemiluminescence assay. This chapter describes these two methods and provides examples of how they have been used successfully to elucidate the role of protein S-nitrosylation in cell physiology and pathophysiology. PMID- 18423222 TI - Pivotal role of arachidonic acid in the regulation of neuronal nitric oxide synthase activity and inducible nitric oxide synthase expression in activated astrocytes. AB - Astrocytes respond to agents leading to progressively greater increases in the intracellular concentration of Ca(2+) ([Ca(2+)](i)) with a linear release of arachidonic acid (ARA), due to activation of cytosolic phospholipase A(2), and with a bell-shaped curve of nitric oxide (NO) release, due to Ca(2+)-dependent activation/inhibition of neuronal NO synthase (nNOS). Inhibition of nNOS is mediated by a signaling driven by ARA, either extensively released at high [Ca(2+)](i) or supplemented to the cultures at nanomolar levels. Proinflammatory factors, as bacterial lipopolysaccharide/interferon-gamma, cause rapid ARA dependent nNOS inhibition, critical for the delayed expression of nuclear factor kappaB (NF-kappaB)-dependent genes as inducible NOS. We therefore propose that the onset of the neuroinflammatory response is strictly regulated by the relative amounts of NO and ARA produced by their constitutive enzymes. In particular, the inflammatory product ARA initiates the inflammatory response via inhibition of nNOS, thereby allowing NF-kappaB activation. Astrocytes contribute to the regulation of this process by producing both constitutive NO and ARA, as well as by expressing NF-kappaB-dependent genes. PMID- 18423223 TI - Red blood cells as a model to differentiate between direct and indirect oxidation pathways of peroxynitrite. AB - Red blood cells are the major physiological scavengers of reactive nitrogen species and have been proposed as real-time biomarkers of some vascular-related diseases. This chapter proposes that the erythrocyte is a suitable cell model for studying the modifications induced by peroxynitrite. Peroxynitrite decays both extra- and intracellularly as a function of cell density and CO(2) concentration, inducing the appearance of distinct cellular biomarkers, as well as the modulation of signaling and metabolism. Intracellular oxidations are due mostly to direct reactions of peroxynitrite with hemoglobin but also lead to the appearance of apoptotic biomarkers. Surface/membrane oxidations are due principally to indirect radical reactions generated by CO(2)-catalyzed peroxynitrite homolysis. PMID- 18423224 TI - Detection and proteomic identification of S-nitrosated proteins in human hepatocytes. AB - The S-nitrosation of protein thiols is a redox-based posttranslational modification that modulates protein function and cell phenotype. Although the detection of S-nitrosated proteins is problematical because of the lability of S nitrosothiols, an increasing range of proteins has been shown to undergo S nitrosation with the improvement of molecular tools. This chapter describes the methodology used to identify potential targets of S-nitrosation in cultured primary human hepatocytes using proteomic approaches. This methodology is based on the biotin switch method, which labels S-nitrosated proteins with an affinity tag, allowing their selective detection and proteomic identification. PMID- 18423225 TI - Identification of S-nitrosylated proteins in plants. AB - Posttranslational protein modifications affect the function or the activity of proteins and exhibit important mechanisms in regulating cellular events. A broad spectrum of modifications is known, including redox-dependent alterations. During the last decade, covalent binding of nitric oxide (NO) to protein cysteines, termed S-nitrosylation, seems especially an evident process for redox-related signaling. To reveal potential target proteins for S-nitrosylation, the biotin switch method gains more and more in importance. This technique is a tool used for analyzing the nitrosylome as well as the examination of single candidates. It is based on substitution of the NO group by a biotin linker that simplifies the detection and the purification of recently S-nitrosylated proteins in a three step procedure. PMID- 18423226 TI - Identification of 3-nitrotyrosine-modified brain proteins by redox proteomics. AB - Two-dimensional (2D) gel electrophoresis allows separation of complex mixtures of proteins based on isoelectric points and relative mobility. This method has not changed much fundamentally since their original description in the late 1970s. Despite several limitations, such as solubilization of membrane proteins and separation of highly basic proteins, this method has been used successfully in many laboratories as part of proteomics protocols. Our laboratory coupled 2D-PAGE with 2D Western blot analysis to identify brain proteins modified oxidatively with excess carbonylation, bound 4-hydroxy-2-nonenal, or 3-nitrotyrosine (3-NT) in various diseases and animal models of these disorders. This chapter describes in detail the protocol used for the identification of 3-NT-modified proteins in biological samples that may help in delineating the role of protein nitration in the progression or pathogenesis of various diseases. PMID- 18423227 TI - Slot-blot analysis of 3-nitrotyrosine-modified brain proteins. AB - 3-Nitrotyrosine (3-NT) is used as a biomarker of nitrosative stress. The formation of 3-NT has been reported in a number of diseases, including Alzheimer's disease, Huntington's disease, Parkinson's disease, amyotrophic lateral sclerosis, and cancer. The nitration of proteins is a reversible process, but it can induce a conformational change and thereby functional alterations of the affected protein. 3-NT measurements in biological samples are usually carried out by methodologies such as immunohistochemistry, high-performance liquid chromatography, gas chromatography, and immunochemical detection. This chapter describes the immunochemical method for the determination of protein-bound 3-NT using slot-blot analysis. PMID- 18423228 TI - Detection assays for determination of mitochondrial nitric oxide synthase activity; advantages and limitations. AB - Nitric oxide (NO) is a reactive radical synthesized by members of the NO synthase (NOS) family, including mitochondrial-specific NOS (mtNOS). Some of the assays used for the determination of cytoplasmic NOS activity have been utilized to detect mtNOS activity. However, it seems that many of those assays need to be adjusted and optimized to detect NO in the unique environment of mitochondria. Additionally, most mtNOS detection assays are designed and optimized for isolated mitochondria and may exert inherent pitfalls and limitations once used in living cells. This chapter describes several assays used commonly for mtNOS detection in isolated mitochondria and in mitochondria of live cells. Those include colorimetric and spectrophotometric methods, Griess reaction, radioassay, and polarographic and chemiluminescence assays. It also describes fluorescent-based assays for the detection of mitochondrial NO in live cells. Advantages and limitations of each assay are discussed. PMID- 18423229 TI - Assay of 3-nitrotyrosine in tissues and body fluids by liquid chromatography with tandem mass spectrometric detection. AB - 3-Nitrotyrosine (3-NT) is a marker of protein nitration in physiological systems. It is present as 3-nitrotyrosine residues in proteins of tissue, extracellular matrix, plasma, and other body fluids and food. It is also present in body fluids and some beverages as free nitrotyrosine and is excreted in urine with the major urinary metabolite 3-nitro-4-hydroxyphenylacetic acid. Quantitation of 3 nitrotyrosine requires tandem mass spectrometry for specific detection. The method developed to determine 3-nitrotyrosine (along with protein glycation and oxidation adducts in a quantitative screening assay) by liquid chromatography with tandem mass spectrometric detection is described. The 3-NT residue contents of plasma protein, hemoglobin, lipoproteins, and cerebrospinal fluid protein and the concentrations of free 3-nitrotyrosine in plasma, urine, and cerebrospinal fluid are given. Changes of 3-nitrotyrosine residue and free 3-nitrotyrosine in diabetes, cirrhosis, acute and chronic renal failure, and neurological disorders, including Alzheimer's disease, are presented and compared with independent estimates. PMID- 18423231 TI - Detection of nitric oxide and its derivatives in human mixed saliva and acidified saliva. AB - Nitrate is secreted into the human oral cavity as a salivary component. The nitrate is transformed to nitrite and nitric oxide (NO) by oral bacteria. NO is oxidized by O(2) producing NO(2) and N(2)O(3) and also by O(2)(-) producing ONOO( ). Salivary peroxidase can oxidize nitrite and NO to NO(2) or its equivalent in the oral cavity. Nitrite dissolved in saliva is mixed with gastric juice, generating nitrous acid that is transformed to NO and NO(2) via N(2)O(3) by self decomposition. In addition, nitrous acid can react with ascorbic acid and phenolics producing NO and with H(2)O(2) producing ONOOH. This chapter deals with the detection of reactive nitrogen oxide species (RNOS), especially NO, N(2)O(3), NO(2), and ONOO(-)/ONOOH, in mixed whole saliva and acidified saliva using fluorescent probes and spin-trapping reagents. It is also shown that measurements of nitration and oxygen consumption are useful in studying the formation and scavenging of RNOS in the aforementioned systems. PMID- 18423230 TI - Nitrite and nitrate measurement by Griess reagent in human plasma: evaluation of interferences and standardization. AB - Nitrite and nitrate represent the final products of nitric oxide (NO) oxidation pathways, and their hematic concentrations are frequently assessed as an index of systemic NO production. However, their intake with food can influence their levels. Nitrite and nitrate could have a role by producing NO, because nitrite can release NO after reaction with deoxyhemoglobin and dietary nitrate can be reduced substantially to nitrite by commensal bacteria in the oral cavity. Different methods have been applied for nitrite/nitrate detection, with the most commonly used being the spectrophotometric assay based on the Griess reagent. However, a reference methodology for these determinations is still missing and many possible interferences have been reported. This chapter assesses how different experimental conditions can influence the results when detecting nitrite and nitrate in human plasma by the Griess assay and provides a simple method characterized by high reproducibility and minimized interferences by plasma constituents. PMID- 18423232 TI - Imaging of reactive oxygen species and nitric oxide in vivo in plant tissues. AB - During the last decades there has been a growing interest in the study of reactive oxygen species (ROS) and nitric oxide (NO) production in plant tissues and their role in signaling and cellular response to biotic and abiotic stress conditions. Despite growing molecular data on this subject, less attention has been paid to the topological distribution of ROS and NO production in plant tissues. Knowledge of the contribution of different cells to the accumulation of ROS and NO is important to get deeper insights into the cellular response of plants to adverse conditions. This chapter focuses on the imaging of ROS and NO accumulation in vivo in plant tissues by confocal laser microscopy using specific fluorescent probes. PMID- 18423234 TI - Male infertility: current concepts and controversies. Preface. PMID- 18423233 TI - Examining nitroxyl in biological systems. AB - Nitroxyl (HNO) has received significant recent attention due to its remarkable and novel biological activity and pharmacological potential. Unlike most other commonly studied nitrogen oxides, examination of HNO biology is not straightforward because of its unique chemical properties. Thus, the study of HNO in biological system requires researchers to be aware of this chemistry in order to properly design and interpret experiments. This chapter focuses on the experimental issues associated with the study of HNO and attempts to provide guidelines for working with this novel and fascinating nitrogen oxide. PMID- 18423235 TI - Declining worldwide sperm counts: disproving a myth. AB - This article explores in detail the issue of the alleged decline in semen quality. The impetus for a comprehensive re-evaluation at this time is threefold: (1) the potential impact of a real decline in semen quality and subsequent human fertility is a priori critical to human welfare; (2) governments have begun to enact "anti-endocrine disruptor" legislation that is based, in part, on selected portions of the published data about semen quality; and (3) confusion and misinformation about semen quality remain widespread in lay and professional circles. PMID- 18423236 TI - Hormonal evaluation of the infertile male: has it evolved? AB - An endocrinologic evaluation of patients who have male-factor infertility has clearly evolved and leads to specific diagnoses and treatment strategies in a large population of infertile men. A well-considered endocrine evaluation is especially essential with the ever-growing popularity of assisted reproductive techniques and continued refinements with intracytoplasmic sperm injection. PMID- 18423237 TI - Assessing sperm function. AB - This article reviews basic semen tests and new fertility tests that are providing great insights to the rapidly developing understanding of male infertility. Finally, promising new tests under development are mentioned with their potential clinical applications. PMID- 18423238 TI - Female fertility: what every urologist must understand. AB - This article provides an overview of the evaluation and treatment of female factor infertility. It reviews the physiology of female reproduction and subsequently discusses pertinent findings in the evaluation and treatment of female infertility that are relevant to a urologist treating the male partner. Finally, it provides an overview of current treatment modalities. PMID- 18423239 TI - Evolving approach to the varicocele. AB - The varicocele is the most common cause of male infertility world wide. Varicoceles have been found in 15% of the normal male population and in up to 40% of patients with male infertility. In approximately 70% of patients with secondary infertility, a varicocele is an underlying cause. Understanding of the pathophysiology, treatments, and outcomes of a varicocele and varicocele repair has evolved significantly over the past several decades. Our goal is to discuss the approach to its diagnosis and treatment that has evolved. PMID- 18423240 TI - Outcomes of varicocelectomy treatment: an updated critical analysis. AB - This article provides an updated analysis of the varicocele literature published since 1994. The present authors have followed the format of the previous review and have included a summary of the results from the 1994 article at the end of each section. PMID- 18423241 TI - Anejaculation and retrograde ejaculation. AB - Anejaculatory infertility is a challenge for the treating physician. Accurate diagnosis and choice of treatment plan must carefully proceed to maximize success rates while maintaining cost-effectiveness. In this article the authors review ejaculatory infertility evaluation and treatment. PMID- 18423242 TI - Ejaculatory duct obstruction. AB - Ejaculatory duct obstruction presents with infertility, pain, or hematospermia. Partial or functional forms of ejaculatory duct obstruction can be difficult to diagnose. Transrectal ultrasound has replaced formal vasography as the first-line diagnostic test but is not specific. Adjunctive procedures such as seminal vesicle aspiration, seminal vesiculography, and chromotubation further delineate the diagnosis. Using an evidence-based approach, this article reviews how best to approach the diagnosis and treatment of ejaculatory duct obstruction. PMID- 18423243 TI - Restructuring reconstructive techniques--advances in reconstructive techniques. AB - Microsurgical reconstruction to correct male infertility, although usually performed for vasectomy reversal, also is performed to correct other types of iatrogenic, congenital, and postinflammatory obstruction. In an effort to improve success rates and facilitate performance of these complex microsurgical procedures, modifications are continually suggested. This article reviews some of these proposed modifications. The modifications can be divided into five general categories: (1) use of biomaterials/sealants, (2) laser soldering, (3) use of absorbable and nonabsorbable stents, (4) new intussusception vasoepididymostomy (VE) anastomotic techniques, and (5) use of robotics. PMID- 18423244 TI - Sperm acquisition in nonobstructive azoospermia: what are the options? AB - Approximately 10% of male-factor infertility is caused by azoospermia, and nearly two thirds of these patients have nonobstructive azoospermia (NOA). As experience has been gained, increasing numbers of men who have NOA are having sperm retrieved from their testes and used for intracytoplasmic sperm injection with vitro fertilization. This article reviews the various sperm retrieval techniques, discussing the advantages and disadvantages and the outcomes of each. Predictive factors for sperm retrieval are presented, as are some of the controversies that exist regarding sperm acquisition in NOA. PMID- 18423245 TI - The genetics you never knew: a genetics primer. AB - Our understanding about genetics is rapidly changing. The goal of this article is to provide an overview of the basics of and new developments in medical genetics- a Genetics 101 primer for 2008. In this article we review the structure and function of the genes, how genes are packaged, gene replication, gene mutations, and the different modes of inheritance. PMID- 18423246 TI - The genetic basis of male reproductive failure. AB - Evolving therapies have allowed the use of sperm from men with spermatogenic compromise, obstructive azoospermia, and sperm functional deficiency, enabling these men to procreate when unable to do so naturally. The genetic basis of only a portion of these conditions is known and research must be pursued into the genetic underpinnings of those that have not yet been delineated. Education and provision of information to patients is the responsibility of all involved in the care of men with reproductive failure. The author concentrates on some of the known causes of nonobstructive azoospermia and obstructive azoospermia with a well-established genetic cause such as congenital bilateral absence of the vas deferens. PMID- 18423247 TI - Intrauterine insemination and male subfertility. AB - In this article we define male subfertility, review clinical indications for the use of intrauterine insemination, evaluate insemination preparation and administration technique, and ultimately advocate for its use as a first-line therapy in the treatment of infertile couples with male subfertility. PMID- 18423248 TI - Intracytoplasmic sperm injection (ICSI)--what are the risks? AB - In vitro fertilization used in combination with intracytoplasmic sperm injection allows otherwise sterile couples to become parents. Despite recent studies on the safety of these technologies, there is still only an incomplete picture of the risks associated with the usage of these assisted reproductive techniques to offspring. The risk of multiple gestations continues to be of major concern because of its association with low birth weight, preterm delivery, and increased perinatal mortality. This article outlines the risks associated with in vitro fertilization/intracytoplasmic sperm injection as a well-defined treatment for couples with severe male factor infertility. PMID- 18423249 TI - Reassessing reconstruction in the management of obstructive azoospermia: reconstruction or sperm acquisition? AB - Treatments for male factor infertility secondary to reconstructable obstructive azoospermia include either surgical reconstruction or direct sperm retrieval. We examine the risks and benefits of both types of therapies and discuss their respective medical and economic implications. Most male factor infertility studies comparing vasectomy reversal with sperm retrieval favor the former as the more cost-effective therapy for obstructive azoospermia. Analysis should include assessment of direct procedural costs and indirect costs, including the cost of complications, lost productivity, and multiple gestation pregnancies. When considering sperm retrieval, the impact of in vitro fertilization-related indirect costs, specifically that driven by multiple gestation pregnancies, is significant. PMID- 18423250 TI - Endocrine manipulation in male infertility. AB - Endocrine therapy for male infertility is broadly categorized as specific or nonspecific therapy. Although uncommon, primary endocrine diagnoses in infertile men are amenable to targeted therapy. The efficacy of empiric endocrine therapy for idiopathic male infertility, however, has not been demonstrated conclusively by clinical trials. With better understanding of the underlying pathophysiology of idiopathic male infertility, careful evaluation of endocrine therapy in well selected treatment groups and well-designed randomized, controlled trials is warranted. Although empiric endocrine therapy for idiopathic male infertility has been largely replaced by assisted reproductive techniques, both treatment modalities could play a role, perhaps as combination therapy. PMID- 18423251 TI - Antioxidant therapy in male infertility. AB - Although reports have suggested the benefit of antioxidant treatment of infertile men, many studies also show no effect. Most studies in the literature are not randomized, placebo controlled, or double blinded in design, which makes it difficult to differentiate regression toward the mean from true positive treatment effects. The small patient sample sizes and varying male populations also add to the difficulty in comparing studies. Pregnancy, the most relevant outcome parameter, is rarely reported. Ideally, patients would be selected based on oxidative stress levels, and improvement in these levels would be correlated to improvement in pregnancy rates. Until those studies are performed, the use of antioxidants for the treatment of male infertility remains empiric. PMID- 18423252 TI - The effect of aging on spermatogenesis and pregnancy outcomes. AB - Until fairly recently, it had been assumed that paternal age had only a minor impact on reproductive outcome. Several recent provocative studies have raised the specter of a causal association between paternal age and significant medical conditions in the offspring. However, the observational nature of these studies leaves open the possibility that factors other than age itself may be responsible for observed results. This article reviews the available data on this topic, with an eye toward providing a basis for clinical counseling of the older man who wishes to have a child. PMID- 18423254 TI - Identity-in-context is not yet identity development-in-context. AB - In this discussion, we summarize the main aspects of the research presented in the papers and discuss what we see as their strong points. To analyze recent developments in identity research, we compared the present issue with the special issue of the Journal of Adolescence in 1996 on 'Identity Development in Context'. In that issue it was concluded that further research is necessary to "detail the ways in which contextual variables may interact both with one another as well as with individual personality variables to determine the course of the identity formation process". In the papers in this issue, the influence of the context has received much more attention than 20 years ago. However, they still have serious limitations with regard to the study of the developmental process. Not more than one study uses a longitudinal design. To stimulate research into developmental processes we conclude with a plea for longitudinal studies over long periods, with enough measurement points to distinguish between different patterns, and for more conceptual attention for what identity development is about. PMID- 18423253 TI - Perceived parenting dimensions and identity styles: exploring the socialization of adolescents' processing of identity-relevant information. AB - This study examined the relationships between crucial dimensions of perceived parenting (support, behavioral control, and psychological control) and the three identity styles defined by Berzonsky [Berzonsky, M. D. (1990). Self-construction over the life span: A process perspective on identity formation. Advances in Personal Construct Psychology, 1, 155-186.]. Each identity style was hypothesized to relate to a specific pattern of perceived parenting dimensions. Hypotheses were examined in a sample of middle and late adolescents (n=674). An information oriented style was positively predicted by parental support. Contrary to expectations, however, an information-oriented style was also positively predicted by psychological control. A normative identity style was positively predicted by support and behavioral control. In line with expectations, a diffuse avoidant identity style was positively predicted by psychological control and negatively by maternal (but not paternal) behavioral control. Findings are discussed in light of the literature on the socialization of identity formation and directions for future research are outlined. PMID- 18423255 TI - Contributions from the 13th International Conference on Bioinorganic Chemistry. Preface. PMID- 18423256 TI - The spectrum of laser skin resurfacing: nonablative, fractional, and ablative laser resurfacing. AB - The drive to attain cosmetic facial enhancement with minimal risk and rapid recovery has inspired the field of nonsurgical skin rejuvenation. Laser resurfacing was introduced in the 1980s with continuous wave carbon dioxide (CO(2)) lasers; however, because of a high rate of side effects, including scarring, short-pulse, high-peak power, and rapidly scanned, focused-beam CO(2) lasers and normal-mode erbium-doped yttrium aluminium garnet lasers were developed, which remove skin in a precisely controlled manner. The prolonged 2 week recovery time and small but significant complication risk prompted the development of non-ablative and, more recently, fractional resurfacing in order to minimize risk and shorten recovery times. Nonablative resurfacing produces dermal thermal injury to improve rhytides and photodamage while preserving the epidermis. Fractional resurfacing thermally ablates microscopic columns of epidermal and dermal tissue in regularly spaced arrays over a fraction of the skin surface. This intermediate approach increases efficacy as compared to nonablative resurfacing, but with faster recovery as compared to ablative resurfacing. Neither nonablative nor fractional resurfacing produces results comparable to ablative laser skin resurfacing, but both have become much more popular than the latter because the risks of treatment are limited in the face of acceptable improvement. LEARNING OBJECTIVES: At the completion of this learning activity, participants should be familiar with the spectrum of lasers and light technologies available for skin resurfacing, published studies of safety and efficacy, indications, methodologies, side effects, complications, and management. PMID- 18423257 TI - Intravenous immunoglobulin selectively decreases circulating autoantibodies in pemphigus. AB - BACKGROUND: Autoantibody-mediated diseases such as pemphigus are caused by a single or very limited number of pathogenic autoantibodies. A major problem with all current therapies for these diseases is that they target all antibodies rather than selectively targeting only pathogenic antibodies. The following study was conducted to confirm observations made in a limited number of patients that suggest intravenous immunoglobulin (IVIg) may be able to selectively lower serum levels of only abnormal autoantibodies. METHODS: The study was conducted in 12 patients who received IVIg for the treatment of recalcitrant pemphigus. Serum levels of antibodies to desmoglein 1 (Dsg 1) and desmoglein 3 (Dsg 3) were measured by enzyme-linked immunosorbent assay immediately before IVIg treatment and following a median of 2 cycles (range, 1-3) of treatment. As control, serum levels of several normal antibodies (against herpes simplex virus types 1 and 2, mumps, and varicella) were measured concurrently. RESULTS: Within a median of 2 weeks following the last cycle of IVIg serum, anti-Dsg 3 declined in all patients who tested positive at baseline and in 8 of 10 (80%) patients testing positive for anti-Dsg 1. On average, anti-Dsg 3 decreased by 45% and anti-Dsg 1 by 32%. By contrast, serum levels of the 4 normal antibodies increased in almost all patients, by an average of 408% (P < .001). LIMITATIONS: Correlation of clinical response to treatment with IVIg was not performed. The sample size was limited. CONCLUSION: These results indicate that IVIg can selectively and markedly decrease serum levels of abnormal antibodies in pemphigus without decreasing the levels of normal antibodies. Thus IVIg appears able to achieve the ideal goal of treatment in autoantibody-mediated diseases--selectively removing from the circulation only those antibodies that cause the disease. PMID- 18423258 TI - Multiple antithrombotic agents increase the risk of postoperative hemorrhage in dermatologic surgery. AB - BACKGROUND: Studies show that holding single antithrombotic agents perioperatively increases the risk of acute thrombotic events and does not significantly decrease the risk of bleeding complications in dermatological surgery. Recent data suggest that selected patients may benefit from combination therapy in preventing acute thrombotic events. OBJECTIVE: We sought to evaluate postoperative bleeding complications in patients who underwent Mohs micrographic surgery while using multiple agents perioperatively compared with patients using a single agent or none at all. METHODS: We conducted a retrospective chart review of patients treated in one academic Mohs micrographic surgery department during 1 year. RESULTS: Patients taking two or more agents at the time of surgery were more likely to bleed than those taking one agent or none at all (P = .0016, Fisher's exact). LIMITATIONS: Small sample size and retrospective nature were limitations. CONCLUSION: Perioperative use of more than one antithrombotic agent increases postoperative bleeding risk. PMID- 18423259 TI - Blood exposure risk during procedural dermatology. AB - BACKGROUND: Dermatologists are at risk of body-fluid contamination during procedures. OBJECTIVE: We sought to determine the frequency of blood splash during procedural dermatology. METHODS: In all, 500 consecutive excisions were performed. Postoperatively, blood droplets on face shields and surgical gowns were counted. A survey regarding universal precautions during procedures was also conducted with members of the American College of Mohs Surgery (ACMS). RESULTS: Contamination from blood splashes during dermatologic procedures (Mohs micrographic surgery, excision, repair) occurred in 66.4%. Reconstruction type, anticoagulation use, wound location, and wound size correlated with a higher blood splash rate. Our survey showed that face shields and goggles are used inconsistently. LIMITATIONS: The 4 participating dermatologists do not represent all practicing dermatologists. It may be possible to generalize the survey results directed at physicians in the ACMS. CONCLUSION: Physician body-fluid contamination risk with procedural dermatology is clinically significant. Dermatologists and their assistants should wear preventive barriers during procedures to minimize the risk of viral transmission. PMID- 18423260 TI - Guidelines of care for the management of psoriasis and psoriatic arthritis: Section 1. Overview of psoriasis and guidelines of care for the treatment of psoriasis with biologics. AB - Psoriasis is a common, chronic, inflammatory, multisystem disease with predominantly skin and joint manifestations affecting approximately 2% of the population. In this first of 5 sections of the guidelines of care for psoriasis, we discuss the classification of psoriasis; associated comorbidities including autoimmune diseases, cardiovascular risk, psychiatric/psychologic issues, and cancer risk; along with assessment tools for skin disease and quality-of-life issues. Finally, we will discuss the safety and efficacy of the biologic treatments used to treat patients with psoriasis. PMID- 18423261 TI - Guidelines of care for the management of psoriasis and psoriatic arthritis: Section 2. Psoriatic arthritis: overview and guidelines of care for treatment with an emphasis on the biologics. AB - Psoriasis is a common, chronic, inflammatory, multisystem disease with predominantly skin and joint manifestations affecting approximately 2% of the population. In this second of 5 sections of the guidelines of care for psoriasis, we give an overview of psoriatic arthritis including its cardinal clinical features, pathogenesis, prognosis, classification, assessment tools used to evaluate psoriatic arthritis, and the approach to treatment. Although patients with mild to moderate psoriatic arthritis may be treated with nonsteroidal anti inflammatory drugs and/or intra-articular steroid injections, the use of disease modifying antirheumatic drugs, particularly methotrexate, along with the biologic agents, are considered the standard of care in patients with more significant psoriatic arthritis. We will discuss the use of disease-modifying antirheumatic drugs and the biologic therapies in the treatment of patients with moderate to severe psoriatic arthritis. PMID- 18423262 TI - The herpes zoster vaccine. PMID- 18423263 TI - Biopsy rates in patients with and without total body photography. PMID- 18423264 TI - Adding over-the-counter dihydroxyacetone self-tanners to sunscreen regimens to increase ultraviolet A light protection. PMID- 18423265 TI - Iatrogenic xanthoderma. PMID- 18423266 TI - Trichoadenoma? [corrected]. PMID- 18423267 TI - Letter to the editor: effects of MTBE on Leydig cell tumors in Sprague-Dawley rats: range of possible poly-3 results. PMID- 18423268 TI - Changes in dental education: dictating the future of the specialty. PMID- 18423270 TI - A method for quantifying condylar motion in patients with osteoarthritis using an electromagnetic tracking device and computed tomography imaging. AB - PURPOSE: The purpose of this study was to develop a method to accurately study the kinematic changes of the temporomandibular joint (TMJ) in patients treated with hemijoint implant reconstruction for dysfunction of advanced degenerative osteoarthritis. MATERIALS AND METHODS: Mandibular kinematic motion data and patient-specific computed tomography (CT) data were acquired. Patients were fitted with custom dental stents that were embedded with metal markers to link the mandibular kinematics data with the 3-dimensional TMJ CT images. An electromagnetic tracking device was used to collect kinematic motion data during maximal mouth opening and closing. The coordinate systems of the kinematic data and CT data were registered to calculate the motion of the mandibular condyle. RESULTS: This technique was successfully used to study patients with motion aberration of the TMJ due to osteoarthritis. A typical case is illustrated in which the motion of both mandibular condyles was simulated preoperatively and postoperatively. CONCLUSIONS: The results of this study suggest that it is possible to use the proposed methodology to accurately quantify the motion of the mandibular condyle in 3 dimensions. The developed technique is user-friendly and noninvasive to the patient. The proposed methodology is a potential clinical tool that may be used in the management of patients with TMJ dysfunction. PMID- 18423269 TI - Inhibition of oral mucosal cell wound healing by bisphosphonates. AB - PURPOSE: Bisphosphonates (BPs) are a widely used class of drugs that are effective in the treatment and prevention of osteoporosis, hypercalcemia of malignancy, and bone metastases associated with multiple myeloma, breast cancer, and other solid tumors. In the past several years there have been numerous reports describing the occurrence of osteonecrosis of the jaws (ONJ) associated with these drugs. Whether the ONJ lesion initiates in the oral mucosa or derives from the underlying bone is not well understood. In this report we describe the effect of pamidronate, a second-generation BP, on oral mucosal cells. MATERIALS AND METHODS: Murine oral keratinocytes were isolated and exposed to pamidronate at a range of clinically relevant doses. Cellular proliferation was measured using a MTS/PMS reagent-based kit and wound healing was examined with a scratch assay. To determine whether oral keratinocytes undergo apoptosis following exposure to pamidronate, TUNEL, caspase-3, and DAPI apoptosis assays were performed. RESULTS: We show that BP pretreatment of oral mucosal cells inhibits proliferation and wound healing at clinically relevant doses, and that this inhibition is not due to cellular apoptosis. CONCLUSIONS: To our knowledge this is the first report investigating the effect of nitrogen-containing BPs on oral mucosal cells. This study suggests that BPs inhibit oral keratinocyte wound healing which may play a significant role in the initiation of ONJ. PMID- 18423271 TI - Office-based ambulatory sedation--the use of the airway protector system during oral surgery: a prospective audit of the first 100 patients. AB - PURPOSE: To evaluate the efficacy of the "Airway Protector System" (APS), a simple homemade device used for airway control during office-based dental sedation. PATIENTS AND METHODS: A prospective audit was performed in 100 severely dental phobic patients submitted to dental treatment in a dentist office under propofol sedation. RESULTS: Sixty-eight females and 32 males were enrolled in this study. The mean age was 45 +/- 7 years (range, 18 to 67 years). A clear airway was obtained in 94 patients. Partial airway obstruction was observed in 4 patients but manual lifting of the jaw was enough to free the partial obstruction and dental treatment was uneventfully conducted. In 2 patients, the APS had to be converted to formal nasotracheal intubation because jaw-lifting maneuvers did not adequately relieve a partial airway obstruction. Sore throat was reported in 56 patients. In 27 cases, the patients rated the complaint as severe. No patient required a chest x-ray after sedation, as there was no clinical evidence of any pulmonary complication including dyspnea, cough, or fever. CONCLUSION: We suggest that the concomitant use of a homemade cuffed nasopharyngeal airway with continuous suction facilitates airway control during deep levels of office-based sedation for dental treatment in severely dental phobic patients. PMID- 18423272 TI - A murine model for septic arthritis of the temporomandibular joint. AB - PURPOSE: This study was designed to establish a murine model of hematogenously acquired bacterial arthritis of the temporomandibular joint (TMJ) to investigate the pathogenesis of this rare TMJ infection. MATERIALS AND METHODS: One hundred forty mice were inoculated intravenously with 3 different staphylococcal suspensions. They were sacrificed at intervals between 1 day and 2 months. Bacterial cultures were obtained from peripheral blood, liver, kidney, TMJs, knees, and interphalangeal joints. The TMJs were collected for histopathological examination. RESULTS: Staphylococcus aureus, isolated from the joint fluid of a patient with septic arthritis of the TMJ, was recovered from the liver, kidneys, knees, interphalangeal joints, and TMJs of several animals. Blood cultures were negative. Acute septic arthritis of the TMJs was confirmed in several animals as soon as 4 days after inoculation. Histopathology showed severe damage to chondrocytes and collagen fibers in the condyles and discs, leading to extensive degenerative changes. All cultures were negative, and there were no histopathologic changes in animals inoculated with bacteria from the other sources. CONCLUSIONS: A murine model for hematogenous septic arthritis of the TMJ was successfully developed with S. aureus isolated from a patient with a TMJ infection. The bacteria induced multiple organ and joint infections. Septic arthritis of the TMJ occurred in 21% of the animals inoculated. Onset was rapid. It produced extensive degenerative changes. The study confirms the need for prompt diagnosis and treatment of patients with septic arthritis. The model may prove to be very useful in the study of this rare infection. PMID- 18423273 TI - Factors related to patients' anxiety before and after oral surgery. AB - PURPOSE: The purpose of this study was to identify factors that may contribute to anxiety of patients undergoing minor oral surgery before and after the operation. PATIENTS AND METHODS: A total of 120 patients who underwent minor oral surgery were included in the study. Each patient's anxiety was measured using Spielberger's State-Trait Anxiety Inventory and Corah's Dental Anxiety Scale immediately before, immediately after, and 1 week after the operation. Anxiety levels for each given situation about the surgery were determined using visual analog scales. The effects of various factors, such as demographic variables, previous dental experience, and postoperative complaints, on anxiety were also analyzed. RESULTS: Postoperative anxiety levels were significantly lower than the preoperative anxiety levels (P<.001). Patients recorded higher anxiety levels for "jaw becoming tired" and "collection of fluid in the mouth" than for "feeling pain during the operation" both before and after the operation. There was a positive correlation between pain expectation with preoperative anxiety and pain actually experienced during surgery with postoperative anxiety (P<.001). Whereas patients' trait anxiety and previous dental experience were correlated with preoperative anxiety (P<.001 and P<.05, respectively), no relationship with postoperative anxiety was found. The difficulty of the procedure was not correlated with anxiety immediately after the operation but was correlated with anxiety during the follow-up visit (P<.05). Postoperative complaints were related to the dental anxiety levels recorded 1 week after surgery (P<.05). CONCLUSIONS: To reduce patients' anxiety, underestimated factors, such as jaw fatigue and fluid collection in the mouth, should be taken into account during oral surgery under local anesthesia. In the postoperative period, swelling and difficulty in eating, as well as pain, should also be considered in relieving patients' anxiety. PMID- 18423274 TI - The effect of informed consent on stress levels associated with extraction of impacted mandibular third molars. AB - PURPOSE: To evaluate the effect of informed consent on stress levels associated with removal of impacted mandibular third molars. PATIENTS AND METHODS: A total of 60 patients scheduled for extraction of impacted mandibular third molars participated in this study. The patients were unaware of the study's objectives. Data from 20 patients established the baseline levels of electrodermal activity (EDA). The remaining 40 patients were randomly assigned into 2 equal groups receiving either a detailed document of informed consent, disclosing the possible risks involved with the surgery, or a simplified version. Pulse, blood pressure, and EDA were monitored before, during, and after completion of the consent document. RESULTS: Changes in EDA, but not in blood pressure, were measured on completion of either version of the consent document. A greater increase in EDA was associated with the detailed version of the consent document (P = .004). A similar concomitant increase (although nonsignificant) in pulse values was monitored on completion of both versions. CONCLUSIONS: Completion of overdisclosed document of informed consent is associated with changes in physiological parameters. The results suggest that overdetailed listing and disclosure before extraction of impacted mandibular third molars can increase patient stress. PMID- 18423275 TI - Effect of estrogen on the expression of cytokines of the temporomandibular joint cartilage cells of the mouse. AB - PURPOSE: The purpose of this study was to evaluate the effect of estrogen on the synthesis of cytokines of the temporomandibular joint cells. METHODS: Mandibular condyles of 10 mice were resected. Twenty of these condylar cartilages were removed and placed in organ culture for 24 hours with media containing different concentrations of 17beta-estradiol. The chondroblasts from the mandibular condyles of 3 mice were cultured with different concentrations of 17beta estradiol the same as above. Cytokine concentrations were measured by ELISA. RESULTS: The expression and concentration of IL-1 beta, IL-6, and IL-8 were increased with increasing concentration of 17beta-estradiol. The expression and concentration of IL-4 and IL-10 were not different from the control and experimental group. CONCLUSIONS: These findings suggest that estrogen has the potential to cause temporomandibular joint disease with induction of the proinflammatory cytokines, IL-1 beta, IL-6, and IL-8. PMID- 18423276 TI - Ocular findings in patients with orbitozygomatic complex fractures: a retrospective study. AB - PURPOSE: To evaluate the incidence of ocular injuries and clinical ocular signs in patients with orbitozygomatic fractures of varying severity, presenting to a regional oral and maxillofacial surgery service. PATIENTS AND METHODS: This is a retrospective study of patients presenting to Limerick Regional Hospital (Limerick, Ireland) with orbitozygomatic fractures from January 1998 to December 2004. Patients with panfacial fractures and those with isolated zygomatic arch fractures were excluded. All patients were examined by the ophthalmology service preoperatively and reviewed postoperatively as indicated. The study population was divided into 3 subgroups based on the extent of the bony injury (confirmed by clinical, plain radiographic, and CT examination) as follows: group 1: "simple" noncomminuted orbitozygomatic complex fractures; group 2: comminuted orbitozygomatic complex fractures; group 3: "pure" orbital blowout fractures. Patient demographics, fracture etiology, and ocular findings were recorded. RESULTS: The study population included 148 patients (130 males, 18 females). All fractures were unilateral. Ocular findings were present in 29 (20%) patients, consisting of 8 of 85 patients (9%) in group 1; 15 of 53 patients (28%) in group 2; and in 6 of 10 patients (60%) in group 3. CONCLUSIONS: Clinical ocular findings and injuries are a relatively common complication of orbitozygomatic fractures, occurring in 29 (20%) patients in this study. These injuries occur more often in patients with orbital blowout fractures compared with comminuted orbitozygomatic complex fractures or simple orbitozygomatic complex fractures. Ophthalmology consultation is recommended for all patients presenting with orbitozygomatic fractures, and is essential for patients with orbital blowout fractures, based on the high incidence of clinical ocular findings and injuries in this subgroup of patients. PMID- 18423277 TI - Evaluation of intraexaminer and interexaminer agreement on classifying lower third molars according to the systems of Pell and Gregory and of Winter. AB - PURPOSE: This study evaluated the intraexaminer and interexaminer reproducibility of 2 systems (Pell and Gregory [P&G] and Winter) for classifying the anatomic position of third molars (M3s) and compared these 2 traditional classification systems with a novel computer-based system (Radio Memory software; Belo Horizonte, Brazil). MATERIALS AND METHODS: Using a descriptive observational study design, we enrolled a sample of 4 surgeons who evaluated M3 position on panoramic radiographs. They classified M3 position (P&G) and inclination (Winter) on 2 different occasions separated by 15 days. M3 position and inclination were also classified using the Radio Memory software. Intraexaminer and interexaminer agreement were computed, as was agreement between the examiners and the Radio Memory software. RESULTS: The radiographic sample comprised 40 M3s. Intraexaminer agreement on classifying the molars according to their inclination was greater than 85% in all cases (mean agreement, 89.37%), whereas reproducibility of classification according to tooth position was observed in only 2 examiners (mean agreement, 66.25%). For the first observations, interexaminer agreement was observed for both classifications, whereas for the second observation, reproducibility among all of the examiners was observed only for the Winter classification. No homogeneity was recorded between the measurements made by the observers and the objective results regarding the position and inclination of the molars determined by the Radio Memory software. CONCLUSIONS: The classification of impacted lower M3s based on their position and following the P&G criteria lacks both intraexaminer and interexaminer reproducibility. However, the levels of intraexaminer and interexaminer agreement are very high when attempting to classify the M3s according to their inclination (Winter classification). These results suggest the advisability of using an objective measurement method to minimize the error introduced by observer interpretation. PMID- 18423278 TI - Skeletal stability after mandibular setback surgery: bicortical fixation using a 2.0-mm locking plate system versus monocortical fixation using a nonlocking plate system. AB - PURPOSE: This study was conducted to compare the time-course changes in condylar long-axis and skeletal stability after sagittal split ramus osteotomy (SSRO) with bicortical plate fixation versus monocortical plate fixation. PATIENTS AND METHODS: Of a group of 40 Japanese patients diagnosed with mandibular prognathism, 20 underwent SSRO with bicortical plate fixation using a locking plate system and the other 20 underwent SSRO with monocortical plate fixation using a conventional plate system. The time-course changes in condylar long-axis and skeletal stability were assessed through axial, frontal, and lateral cephalograms. RESULTS: Significant differences were found between the 2 groups in changes of the left condylar angle between the initial and 1-month measurements (P = .0454) and in ANB between the 1- and 3-month measurements (P = .0206); however, no significant differences were found between the 2 groups in the other measurements in each time interval. CONCLUSIONS: Our findings suggest no significant differences in postoperative time-course changes between bicortical plate fixation using a locking plate system and monocortical plate fixation using a conventional plate system. PMID- 18423279 TI - Histomorphometric evaluation of the effects of zoledronic acid on mandibular distraction osteogenesis in rabbits. AB - PURPOSE: To assess the effect of systemic administration of zoledronic acid (ZA) on mineralization of newly formed bone and to determine strain-related osteoporosis on surrounding bone during lengthening of immature rabbit mandible. MATERIALS AND METHODS: Eighteen New Zealand white rabbits were divided randomly into 2 groups, and bone lengthening was carried out in the left portion of the mandible through distraction osteogenesis with a rate of 0.5 mm every 12 hours for 5 days. The experimental group was administered 0.1 mg/kg ZA intravenously. The control group was given saline infusion only during operation. All animals were sacrificed at the end of the 28-day consolidation period. The mandibles of all animals were removed and regenerate was evaluated. Osteoblasts, osteoclasts, collagen fibers, and fibroblasts were marked within 0.1-mm(2) area and newly formed bone area was measured within 0.5-mm(2) area. All data were analyzed using Mann-Whitney U test. RESULTS: Although irregular bone destruction spots were seen in the control group, the experimental group showed regular ossification areas and significant difference between osteoblast and osteoclast numbers (P < .05). In the regenerate zone, there was considerable difference between the 2 groups in terms of osteoblast, osteoclast, and collagen amounts (P < .05). Additionally, newly formed bone areas and fibroblast count were higher in experimental group. CONCLUSIONS: The results of this study showed that ZA had positive effects on the new bone formation, which may potentially shorten the consolidation period. PMID- 18423280 TI - Pain experience after simple tooth extraction. AB - PURPOSE: To assess pain experience after simple uncomplicated tooth extraction and to see if there is a need to prescribe analgesic drugs after such a procedure. PATIENTS AND METHODS: A random sample of patients presenting for tooth extraction at the Maxillofacial Unit, Jordan University of Science and Technology was included. A baseline assessment of previously experienced general and dental pains using numeric scales was done. Subsequently, tooth extractions were done and telephone interviews were made during evenings for a week. Pain intensity was assessed on a numeric scale, and use of analgesic drugs and pain quality were recorded. RESULTS: At the evening of extraction 81.8% of patients had pain. Female gender predominance in pain reporting was statistically significant on postextraction days 3 and 5. Chronically inflamed teeth caused the highest mean pain intensity scores and nonsmokers showed significantly higher mean pain intensity scores compared with smokers. Mild pain was experienced by most patients (38.6%) on the evening of extraction. It was found that 55.3% of participants (largely females) used analgesic drugs on the evening of extraction, and 6.8% of participants still used analgesic drugs on day 7 postextraction. There was a significant correlation between mean pain intensity score and previous dental injection pain. CONCLUSION: Patients, notably females, experienced pain of varying intensity after simple uncomplicated tooth extraction maximally at the evening of extraction; and greater than 50% of the patients used analgesic drugs. We recommend offering regular analgesic drugs during the first week after tooth extraction. PMID- 18423281 TI - Characterization of gene expression profiles of 3 different human oral squamous cell carcinoma cell lines with different invasion and metastatic capacities. AB - PURPOSE: The gene expression of 3 oral squamous cell carcinoma (OSCC) human cell lines, BHY, HN, and HSC-3, were studied based on their reported ability to invade adjacent bone or metastasize to cervical lymph nodes and/or distant organs. MATERIALS AND METHODS: The characteristics of each cell line were confirmed on scid mice using micro-positron emission tomography (PET)/computerized tomography (CT) imaging techniques. Complimentary DNA (cDNA) microarray techniques were used to determine the gene expression profile differences between each of the three OSCC cell lines. RESULTS: BHY, HN, and HSC-3 cell lines expressed 139, 214, and 128 up-regulated genes; and 117, 262, and 117 down-regulated genes, respectively. The clusterization of data showed that there are 13 genes that are up-regulated and 83 genes that are down-regulated in all 3 OSCC cell lines. Collection of genes organized by pathway may cause aggregate evaluation of anomalies. Thus the pathway analysis performed for each cell line based on cDNA microarray results showed BHY, HN, and HSC-3 cell lines to have 8, 10, and 3 up-regulated pathways and 3, 9, and 6 down-regulated pathways, respectively. CONCLUSIONS: This study showed that cDNA microarray analysis is an effective tool for mapping molecular signatures. With this technique it is possible to observe the entire genome of a malignant tumor so as to appreciate the simultaneous interactions among thousands of genes. PMID- 18423282 TI - Immunohistochemical evaluation of giant cell tumors of the jaws using CD34 density analysis. AB - PURPOSE: To compare CD34 expression in both aggressive and nonaggressive giant cell lesions of the jaws and identify any associations between tumor vascular density and biologic behavior. MATERIALS AND METHODS: This was a retrospective study of subjects treated for giant cell lesions of the jaws at Massachusetts General Hospital from 1992 to 2006. The primary predictor variable was tumor classification (aggressive or nonaggressive); tumors were considered aggressive if they were greater than 5 cm in size, recurred after treatment, or exhibited 3 of the following: presence of root resorption, tooth displacement, or cortical bone thinning or perforation. Secondary predictor variables, recorded for each patient, were demographic, anatomic, and clinical measures. The outcome variable was the average CD34 staining density of histologic specimens quantified in 2 different areas. Descriptive and bivariate statistics were computed to identify predictors associated with vascular density. RESULTS: The study sample was composed of 32 subjects with a mean age of 24.4 +/- 19.77 years (range: 2-83); 23 subjects (71.8%) were female. Of the tumors included, 11 (34.4%) were located in the maxilla, 21 (65.6%) in the mandible. Twenty-six tumors (81.2%) were classified as aggressive; the remainder (18.8%) were nonaggressive. There were no statistically significant differences between subjects with aggressive versus nonaggressive tumors with regard to age, gender, or location. Subjects with aggressive tumors had a significantly higher CD34 staining density (P = .02). None of the secondary predictors was associated with vascular density. CONCLUSION: Vascular density of giant cell tumors of the jaws is significantly increased in aggressive tumors. PMID- 18423283 TI - Complex orthognathic surgery: assessment of patient satisfaction. AB - PURPOSE: The purpose of our survey study was to determine if bimaxillary orthognathic surgery with simultaneous intranasal surgery and other procedures carried out primarily for the correction of a developmental dentofacial deformity can be completed with a high level of patient satisfaction and minimal complications. PATIENTS AND METHODS: A consecutive series of 42 patients entered in the study from senior surgeon's (J.P.) patients at a single institution who underwent the minimum designated simultaneous procedures (Le Fort I, sagittal osteotomies of the mandible, septoplasty, inferior turbinate reduction) during a 1-year period. Patient satisfaction was assessed through analysis of data gathered from a postsurgical patient satisfaction questionnaire. The questionnaire is used to assess overall postsurgical/orthodontic patient satisfaction and patient assessment of head and neck function. The questionnaire was independently completed by each subject at least 6 months after surgery and only after removal of all orthodontic appliances and planned dental rehabilitation. RESULTS: The results of our study clarify that bimaxillary orthognathic surgery including simultaneous intranasal (septoplasty and turbinate reduction) and other procedures (genioplasty, liposuction, and removal of third molars) can be carried out with a high level of patient satisfaction (89% of our study patients). Nevertheless, 2 of 42 patients (5%) in our study group were dissatisfied despite the absence of surgical or orthodontic complications and the clinicians' feelings that the results achieved were an improvement. CONCLUSIONS: The results of our study clarify that complex bimaxillary orthognathic surgery including simultaneous intranasal and other procedures can be carried out with a high level of patient satisfaction. PMID- 18423284 TI - Treatment of delayed-onset infections after impacted lower third molar extraction. AB - PURPOSE: To describe the treatment of delayed-onset infections after lower third molar removal. PATIENTS AND METHODS: A retrospective study was made of 33 delayed onset infections after impacted lower third molar extraction diagnosed between 2001 and 2005 in the Oral Surgery and Implantology Department of the School of Dentistry of the University of Barcelona, Spain. RESULTS: Antibiotic treatment was effective in 22 patients, whereas the remaining 11 needed an additional surgical procedure to resolve this postoperative complication. Patients with prolonged use of antibiotics after the onset of the infection were more likely to require surgical intervention. CONCLUSIONS: Patients with delayed-onset infections should be treated initially with antibiotics for 7 days. If the infection does not respond favorably within that time, surgical debridement of the extraction site should be done. PMID- 18423285 TI - Caries experience and periodontal pathology in erupting third molars. AB - PURPOSE: This study was conducted to document the prevalence of occlusal caries experience and periodontal pathology for erupting third molars in young adults. PATIENTS AND METHODS: The data are from 49 subjects enrolled in an institutional review board-approved trial with at least one third molar below the occlusal plane at baseline that erupted by longest follow-up. Teeth were considered erupted if they reached the occlusal plane. Caries experience on the occlusal surface of third molars was assessed by a visual-tactile examination. At least 1 periodontal probing depth (PD) >or=4 mm in the third molar region was considered indicative of periodontal pathology. The third molar region was defined as the 6 probing sites around third molars and 2 sites on the distal of second molars. The prevalence of third molar caries experience and periodontal pathology at longest follow-up was assessed. RESULTS: Most of the 49 subjects were female (51%), Caucasian (76%), and educated at least through high school (82%). Median age was 20.5 years (interquartile range [IQR] 18.4 to 24.1 years). Median follow-up was 5.1 years (IQR = 3.4 to 6.9 years). At baseline, none of the subjects had occlusal caries experience in a third molar; 51% of subjects had at least 1 PD >or=4 mm in a third molar region. At follow-up, 27% of the subjects had occlusal caries experience in at least 1 third molar that erupted to the occlusal plane; 61% had at least 1 PD >or=4 mm in a third molar region. Twenty-nine percent had occlusal caries in at least 1 third molar at the occlusal plane and at least 1 PD >or=4 mm in a third molar region. Thirty-seven percent had no third molar occlusal caries experience and all third molar region PD <4 mm. CONCLUSIONS: For third molars that erupted "late," periodontal pathology was more prevalent than occlusal caries. PMID- 18423286 TI - Interventional sialoendoscopy: early clinical results. AB - PURPOSE: Sialoendoscopy is a novel minimally invasive technique to explore the salivary duct system and to treat obstructive salivary disease. This article describes the early clinical experience with endoscopic salivary duct exploration and sialolithectomy in 2 medical centers. PATIENTS AND METHODS: This is a retrospective case series of 94 patients, with submandibular (n = 77) or parotid (n = 17) sialadenitis secondary to sialolithiasis, strictures, or mucous plugs. Patients underwent sialoendoscopy at Baptist Hospital, Miami (n = 52) or at Massachusetts General Hospital, Boston (n = 42). Dilatation of the duct through the natural orifice was accomplished with salivary dilators. Three endoscope systems with diameters from 1.1 to 2.3 mm were used. Using a basket, grasper, lithotripsy, laser, or a combination of these, stones were fragmented or removed endoscopically. Strictures were dilated and mucous plugs removed. All cases were carried out under general anesthesia. RESULTS: Salivary duct navigation was accomplished in 91/94 patients. In 3 cases, duct dilatation was not possible due to scarring. Symptomatic relief was achieved in 81/91 patients (89.4%). Strictures and mucous plugs were visualized and managed in 18/18 patients. Sialoliths were visualized in 73 patients and stone fragmentation or retrieval was accomplished in 84.93% (62/73) of cases. Complications included 2 patients with temporary lingual nerve paresthesia and 1 patient with excess extravasation of irrigation fluid. CONCLUSION: The results of this study indicate that interventional sialoendoscopy is an effective, minimally invasive alternative treatment for obstructive salivary gland disease. PMID- 18423287 TI - Facial trauma coverage among level-1 trauma centers of the United States. AB - PURPOSE: A large portion of patients admitted to trauma centers present with isolated or concomitant facial injuries. Multiple surgical specialties including oral and maxillofacial, plastic, and otolaryngology/head and neck surgeons are trained and involved in the management of oral and maxillofacial trauma. The purpose of this study is to evaluate the current distribution of different specialties that cover facial trauma among the leading trauma centers in the United States. MATERIALS AND METHODS: Based on the number of emergency department admissions, the 5 busiest hospitals for each state within the United States were determined from the American Hospital Association, representing a total of 255 hospital emergency rooms. Seventy-six (N = 76) level-1 trauma centers were identified by directly contacting the hospital facility. Information was requested by mail from the chief of surgery and emergency medicine department regarding the percentage of facial trauma coverage by the different surgical specialties. In addition, the professional societies representing the 3 main specialties: American Association of Oral and Maxillofacial Surgeons (AAOMS), American Association of Plastic Surgeons (AAPS), and the American Academy of Otolaryngologists/Head and Neck Surgeons (AAO-HNS) were contacted to determine an estimate of the number of active members within each specialty. RESULTS: Information was obtained from 57 (75%) level-1 trauma hospitals. The remaining 19 (25%) hospitals failed to respond to our survey. The distribution of facial trauma coverage by the different specialties was as follows: plastic surgeons, 39.6%; oral and maxillofacial surgeons, 36.6%; otolaryngologists/head and neck surgeons, 23.3%; and other services (general surgery and oculoplastics), 0.5%. According to the respective professional societies contacted, there are approximately 7,003 plastic surgeons, 6,377 oral and maxillofacial surgeons, and 7,720 head and neck surgeons that are practicing members of their respective societies. CONCLUSION: Treatment of facial trauma is an essential and demanding aspect of all the surgical specialties that provide this service. All major trauma centers require the support of facial trauma specialists for management of these injuries. When considering the ratio of surgeons per specialty and the percentage of facial trauma coverage provided by each specialty, oral and maxillofacial surgeons and plastic surgeons provide the greatest proportion of facial trauma coverage among the level-1 trauma centers that participated in the survey. PMID- 18423288 TI - Quality of life evaluation of maxillomandibular advancement surgery for treatment of obstructive sleep apnea. AB - PURPOSE: The purpose of this study was to investigate the quality of life (QOL) for patients who underwent maxillomandibular advancement (MMA) surgery for obstructive sleep apnea (OSA) treatment using the Functional Outcomes of Sleep Questionnaire (FOSQ). Correlation of the findings with parameters from the polysomnograph (PSG) and physical examination were also performed. No QOL studies have been carried out for MMA in OSA, and the FOSQ is the most appropriate method. PATIENTS AND METHODS: The FOSQ instrument is a survey designed to evaluate QOL in OSA and proven to be effective. All patients followed a standard prospective protocol of pre- and 6 months postoperative data collection. Clinical radiographs, PSG, FOSQ, endoscopy, and computed tomography scans were analyzed. RESULTS: Fifteen patients with complete data were included in the study. There was minimal change in the BMI from a preoperation mean of 32.10 to 31.50 at 6 months postoperation. The operation was 86.7% (13 of 15) successful (postoperative apnea and hypopnea index [AHI] decreased by 50% and was <20), with a significant decrease of the mean preoperation apnea-hypopnea index (AHI) of 69.12 to 13.87 postoperation (P < .0001). The minimum oxygen saturation (Lsat) correspondingly increased from a mean of 76.49 to 84.96 (P = .0001). A total of 93.3% of our patients achieved a successful QOL change based on a FOSQ score of greater than or equal to 18. There was statistically significant changes in all domains; general productivity (P = .0003), social outcome (P = .0020), activity level (P = .0008), vigilance (P = .0028), intimacy and sex (P = .0039), and the total score (P = .0002) postoperatively. Changes of Lsat and AHI were highly correlated (r = -0.728, P = .002). The magnitude of the Le Fort advancement was also found to be negatively correlated to the change in AHI (r = -0.544, P =.036). There was no correlation between the FOSQ scores and all other parameters, including both the PSG parameters (AHI and Lsat). CONCLUSION: MMA remains the most effective operation for the treatment of OSA and yet no QOL studies have been carried out. The disease-specific QOL instrument (FOSQ) showed the subjective functional improvement in MMA patients. QOL instruments should be used for all surgical based treatment as they add a new dimension to the assessment of the patient and procedures. PMID- 18423289 TI - Evidence-based oral and maxillofacial surgery. AB - PURPOSE: The amount and quality of research evidence in oral and maxillofacial surgery (OMFS) journals have never been evaluated. The current study aims to empirically assess the evidence available in this literature. MATERIALS AND METHODS: The main (Database 1) and neighboring (Database 2) journals of OMFS were manually screened over a 3-year period (2004 to 2006). The types and designs of articles were recorded. The identified randomized, controlled trials (RCTs) were further evaluated for issues of reported methodological quality. RESULTS: In total, 3,487 articles were analyzed. Meta-analyses and RCTs were very rare. Only 2 meta-analyses of RCTs were identified, and RCTs represented a mere 1.3% in each database. Case series and case reports were the dominant article types (1,388, or 59%, for Database 1; 686, or 60.1%, for Database 2). Basic research, laboratory and animal studies, had an important share of 19% and 15.4% in both databases, respectively. An equally high percentage was recorded for nonsystematic reviews, personal views, expert opinions, and editorials (15.9% and 19% for Database 1 and Database 2, respectively). Of the 46 identified RCTs, most of them enrolled less than 100 patients (37 studies, or 80.5%). The majority of them did not describe the randomization mode (27 studies, or 59%), did not present power calculations (31 studies, or 67.4%), and did not report allocation concealment (38 studies, or 79.5%). Almost half of them made no mention of masking (22 studies, or 48%), and only one third described withdrawals during follow-up (15 studies, or 32.6%). CONCLUSION: The OMFS literature suffers from a relative shortage of high-quality evidence. More, larger, adequately powered, and better reported RCTs are warranted. PMID- 18423290 TI - Mandible matrix necrosis in beagle dogs after 3 years of daily oral bisphosphonate treatment. AB - PURPOSE: An increasing number of reports have implicated bisphosphonates as contributing to osteonecrosis of the jaw. The goal of this study was to evaluate mandible necrosis in beagle dogs treated for 3 years with oral alendronate (ALN). MATERIALS AND METHODS: Skeletally mature female beagles were treated daily for 3 years with oral doses of vehicle (VEH) or ALN (0.20 or 1.0 mg/kg/day). These doses approximate, on a mg/kg basis, those used for postmenopausal osteoporosis and Paget's disease, respectively. At necropsy, the second molar region of the mandible was excised, stained en bloc with basic fuchsin, and assessed for matrix necrosis and intracortical bone turnover rate using histology. Matrix necrosis was defined as a region greater than 500 microm(2) that was void of basic fuchsin stain, assessed using both bright-field and confocal microscopy. RESULTS: No animals developed exposed bone lesions in the oral cavity during the 3-year study. Matrix necrosis was observed in 25% of ALN0.2 animals, 33% of ALN1.0 animals, and was noticeably absent from all vehicle animals (P < .05 pooled ALN doses vs VEH). These necrotic regions occurred predominately in the alveolar bone and were clearly void of patent canaliculi. Intracortical bone turnover rate of the alveolar mandible bone region was significantly lower (-75%, P < .05) in ALN treated animals compared with VEH. CONCLUSIONS: Three years of daily oral bisphosphonate treatment reduces bone turnover significantly and increases the incidence of matrix necrosis within the mandible of dogs. PMID- 18423291 TI - Mental neuropathy as a manifestation associated with malignant processes: its significance in relation to patient survival. AB - PURPOSE: Mental neuropathy (MN) is characterized by the presence of a sensory defect in the form of paresthesias or dysesthesias in the territory innervated by the mental nerve. MN may be the first manifestation of systemic cancer, a symptom of spread of an established tumor, or a sign of infiltration in an intraoral lesion. In any of these cases, the symptom is indicative of a very poor patient prognosis. MATERIALS AND METHODS: A total of 22 cancer patients with chin paresthesia were studied. Group 1 comprised patients with chin paresthesia who had a primary tumor in some other region at a distance from the oral cavity or maxillofacial zone. Group 2 in turn comprised patients with primary malignancies of the oral and/or maxillofacial territory and who likewise presented with chin paresthesia. Data were collected relating to patient age, gender, primary intraoral lesion (location, size, histologic diagnosis), primary systemic tumor, and mean patient survival. RESULTS: Group 1 consisted of 11 patients (8 men and 3 women), aged between 36 and 81 years (mean, 58.09 +/- 14.99 years), with different systemic cancers. The mean survival after the diagnosis of chin paresthesia was 14.8 +/- 16.5 months, and only 1 patient was still alive after 9 months. Group 2 consisted of 11 patients (8 men and 3 women), aged between 33 and 72 years (mean, 56.18 +/- 15.69 years). All presented with oral squamous cell carcinoma, with the single exception of 1 case of fibrosarcoma. In this group the mean survival of the 8 patients who died was 28.2 +/- 29.6 months. Three patients survived for a mean of 17 months. CONCLUSIONS: Chin paresthesia is a very important prognostic symptom determining the degree of infiltration of intraoral lesions, and in some cases it may be indicative of the existence of a primary tumor (identified or otherwise), with poor short-term survival--given that 81.9% of the patients studied (18 cases) had died before a mean of 20 months. Although mean survival was shorter (14.8 months) among the patients in group 1 than in group 2 (28.2 months), the difference was not statistically significant. PMID- 18423292 TI - Fibula free flap with endosseous implants for reconstructing a resected mandible in bisphosphonate osteonecrosis. AB - PURPOSE: Bisphosphonates are a class of drugs with several uses. Primarily, they are used to treat bone metastases, multiple myeloma, Paget's disease, and osteoporosis. However, the use of bisphosphonates can lead to osteonecrosis of the jaws. Here, we present our experience treating this widely discussed disease. PATIENT AND METHODS: We describe a patient who underwent surgical reconstruction of the mandible with a fibula free flap after total mandibulectomy, performed to treat osteonecrosis induced by the use of intravenous bisphosphonates complicated by a mandibular fracture. Endosseous implants were inserted simultaneously to optimize the length and outcome of surgery. RESULTS: Osteonecrosis of the jaws is a treatment challenge, and several approaches have been suggested. In this patient, the use of a fibula free flap to reconstruct the mandible was a winning strategy. The patient is now free from disease, and the esthetic results were optimal. CONCLUSIONS: With the simultaneous placement of endosseous dental implants, the functional outcome was optimal, with no significant complications or recurrence. PMID- 18423293 TI - The influence of tobacco on early dental implant failure. AB - PURPOSE: The purpose of this study was to assess tobacco use as a risk factor for early implant failure. MATERIALS AND METHODS: To address the research aim, a retrospective study design was used, and a study sample was derived from among patients who had 1 or more implants inserted between July 1996 and July 2004. The main predictor variable was the frequency of tobacco use. The major outcome variable was early implant failure. Appropriate descriptive, bivariate, and multivariate statistics were computed. RESULTS: The study sample was composed of 650 patients who had 1,628 implants inserted and were selected for analysis. The early implant loss rates found in the nonsmoking group and the smoking group were 3.32% and 2.81%, respectively. Univariate and bivariate analyses showed no statistical significance for early implant losses associated with the frequency of tobacco use. CONCLUSIONS: The results of this study suggest that tobacco use alone cannot be considered as a factor for risk related to early implant failures. Prospective studies are needed to assess the risk of early implant failure in conjunction with smoking. PMID- 18423294 TI - The book flap: a technical note. AB - PURPOSE: To present a useful technique for widening the thin alveolar ridge for placement of an implant or bone graft. MATERIALS AND METHODS: A partial soft tissue flap is elevated over the alveolar ridge and the ridge is split with an osteotome. The technique out-fractures the buccal plate and maintains the labial soft tissue to preserve the blood supply to the buccal bone. RESULTS: The book flap has been employed in over 500 cases over the past 12 years. Approximately 90% were successful in providing adequate width to accommodate an implant. Infection occurred in approximately 2% of cases. The average gain in width was around 3 mm. CONCLUSION: This technique can be employed often and has a very low complication rate. PMID- 18423295 TI - Flapless implant surgery: review of the literature and report of 2 cases with computer-guided surgical approach. AB - PURPOSE: Modern implantology uses techniques that can provide function, esthetics, and comfort with a minimally invasive surgical approach. Flapless implant surgery has been proposed to fulfill these requirements. Traditionally, flapless implant surgery was carried out by using a tissue punch technique, which may be potentially harmful because of the inherent blindness of the technique. The purpose of this article is to introduce a predictable flapless approach for treatment of 2 patients through principles of computer-guided implantology. MATERIALS AND METHODS: Using dedicated interactive computer software programs and 3D radiographic techniques such as computed tomography (CT), the precise location of each implant was planned. Using the concept "prosthetic-driven implantology," surgery was carried out for the rehabilitation of 2 patients. RESULTS: The procedure of implant planning in this sophisticated technique has potential to yield substantial public health benefits. CONCLUSION: Although this modern approach may have many advantages, transferring virtual planning to the patient's mouth has only been developed recently. Further research is required to justify this novel approach for implant rehabilitation. PMID- 18423296 TI - Oral bisphosphonates and dental implants: a retrospective study. AB - PURPOSE: The objective of this retrospective study was to examine whether patients who take medications containing bisphosphonates (BPs) are at greater risk of bone graft and implant failure than other patients. PATIENTS AND METHODS: This study involved the examination of 42 patients (101 implants) who had taken medications containing BPs prior to surgeries involving oral bone grafting or endosseous implant placement. Patients had been taking BP medications from 6 months to 11 years prior to implant surgery, and most continue to take these medications through the present time. Patients were examined to determine implant loss, changes in pocket depth bleeding on probing, the height of the ridge, and any evidence of osteonecrosis of the jaw. RESULTS: Five implants failed, giving a 95% success rate, which is comparable to the normal success rate of 96.5% by the same operator. No patient showed signs of osteonecrosis of the jaw. CONCLUSION: Patients who take oral BPs are no more at risk of implant or bone graft failure than other patients. PMID- 18423297 TI - Surgical treatment algorithm for odontogenic keratocyst: combined treatment of odontogenic keratocyst and mandibular defect with marsupialization, enucleation, iliac crest bone graft, and dental implants. PMID- 18423298 TI - The past, present, and future of oral and maxillofacial surgery. PMID- 18423299 TI - Esophageal rupture with the use of the Combitube: report of a case and review of the literature. PMID- 18423300 TI - A combined orthodontic and surgical approach in osteogenesis imperfecta and severe Class III malocclusion: case report. PMID- 18423301 TI - Synchronous Warthin tumors and lymphoepithelial cyst in the ipsilateral parotid gland. PMID- 18423302 TI - Pediatric maxillary odontogenic myxoma: a report of 2 cases and review of management. PMID- 18423303 TI - Peripheral giant cell granuloma associated with hyperparathyroidism secondary to end-stage renal disease: a case report. PMID- 18423304 TI - Cervical Castleman's disease in childhood. PMID- 18423305 TI - Multicentric Gorham's disease in the oral and maxillofacial region: report of a case and review of the literature. PMID- 18423306 TI - Marsupialization catheter. PMID- 18423307 TI - Immediate implants placed into infected sockets. PMID- 18423308 TI - Aloha to a solution looking for a problem. PMID- 18423309 TI - Abstracts of the XIV International Congress on Nutrition and Metabolism in Renal Disease. Marseilles, France. June 11-15, 2008. PMID- 18423311 TI - Radiology resident interpretation of on-call CT pulmonary angiograms. AB - RATIONALE AND OBJECTIVES: To evaluate the interpretation of computed tomographic pulmonary angiograms performed outside of regular reporting hours, comparing the initial interpretation by the radiology resident to the attending radiologist. MATERIALS AND METHODS: Records for 840 consecutive computed tomographic pulmonary angiograms (CTPA) performed outside of regular reporting hours at two tertiary referral centers from January 1, 2004-December 31, 2005 were reviewed. The preliminary interpretation by the on-call radiology resident was compared to the subsequent final report issued by a subspecialty trained chest radiologist. Studies were stratified as positive, negative, or equivocal for pulmonary embolus. Cases with discordant interpretations or negative CTPA were reviewed to determine impact on clinical outcome. Patients were followed up to 12 months after CTPA to document any subsequent thromboembolic event. RESULTS: Sixteen percent (131/840) of CTPAs were reported positive by the staff radiologist. There was agreement in 90% (752/840) of studies (P = .76, 95% confidence interval, 0.71 0.81) with 86% (114/133) agreement for studies interpreted as positive by residents, 95% (582/612) for studies interpreted as negative by residents, and 63% (60/95) for studies interpreted as equivocal by residents. Studies of optimal quality had higher interobserver agreement than studies of suboptimal quality (P < .0001). In-patient studies were more likely to be positive than emergency room patients (20% vs. 13%) (P = .004). No adverse clinical outcomes were attributed to discordant interpretations. CONCLUSIONS: Radiology residents provide a high level interpretation of on-call CTPA studies, achieving good concordance with the attending radiologists' assessment. PMID- 18423312 TI - Pharmacokinetic analysis of malignant pleural mesothelioma-initial results of tumor microcirculation and its correlation to microvessel density (CD-34). AB - RATIONALE AND OBJECTIVES: Malignant mesothelioma (MM) of the pleura is an aggressive and often fatal neoplasm. Because MM frequently demonstrates marked angiogenesis, it may be responsive to antiangiogenic therapy, but effective methods for selecting and monitoring of patients are further needed. We employed dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) and quantitative immunohistochemistry (IHC) to characterize the microvascularity of MM using both a physiologic and ultrastructural method. MATERIALS AND METHODS: Nineteen patients diagnosed with MM were enrolled and DCE-MRI was performed before antiangiogenic treatment. For each patient, tumor regions were characterized by their DCE-MRI-derived pharmacokinetic parameters (Amp, k(ep), k(el)), which were also compared to those of normal tissue (aorta, liver, spleen, and muscle). In addition, quantitative IHC of representative samples was performed with CD-34 staining to compare the calculated microvessel density (MVD) results with DCE-MRI results. RESULTS: MM demonstrated markedly abnormal pharmacokinetic properties compared with normal tissues. Among the parameters tested, Amp was significantly different in MM (P < or = .001) compared to normal organs. Despite the observation that the MVD of mesotheliomas in this series was high compared to other tumors, DCE-MRI pharmacokinetic parameters had a moderately positive correlation with MVD (r = 0.5). CONCLUSIONS: DCE-MRI and IHC can be used in patients with MM to visualize tumor microvascularity and to characterize tumor heterogeneity. DCE-MRI and IHC results positively correlated, though moderately, but these two methods present as essential tumor biomarkers. This multimodal characterization may be useful in selecting possible tumor subtypes that would benefit from antiangiogenic therapy. PMID- 18423310 TI - Computer-aided diagnosis of lung cancer and pulmonary embolism in computed tomography-a review. AB - Computer-aided detection (CADe) and computer-aided diagnosis (CADx) have been important areas of research in the last two decades. Significant progress has been made in the area of breast cancer detection, and CAD techniques are being developed in many other areas. Recent advances in multidetector row computed tomography have made it an increasingly common modality for imaging of lung diseases. A thoracic examination using thin-section computed tomography contains hundreds of images. Detection of lung cancer and pulmonary embolism on computed tomographic (CT) examinations are demanding tasks for radiologists because they have to search for abnormalities in a large number of images, and the lesions can be subtle. If successfully developed, CAD can be a useful second opinion to radiologists in thoracic CT interpretation. In this review, we summarize the studies that have been reported in these areas, discuss some challenges in the development of CAD, and identify areas that deserve particular attention in future research. PMID- 18423313 TI - Evaluation of a real-time interactive pulmonary nodule analysis system on chest digital radiographic images: a prospective study. AB - RATIONALE AND OBJECTIVES: We sought to assess the performance of a real-time interactive pulmonary nodule analysis system for evaluation of chest digital radiographic (DR) images in a routine clinical environment. MATERIALS AND METHODS: A real-time interactive pulmonary nodule analysis system for chest DR image softcopy reading (IQQA-Chest; EDDA Technology, Princeton Junction, NJ) was used in daily practice with a Picture Archiving and Communication System in a National Cancer Institute-designated cancer teaching hospital. Patients referred for follow-up of known cancer underwent digital chest radiography. Posteroanterior and lateral DR images were first read by resident radiologists along with experienced chest radiologists using a Picture Archiving and Communication System work station. The computer-assisted detection (CAD) program was subsequently applied to the posteroanterior DR images, and changes (if any) in diagnosis were recorded. For reference standard, a follow-up chest radiograph at least 6 months following the initial examination or a follow-up computed tomographic scan of the chest within 3 months was used to establish diagnostic accuracy. RESULTS: Of 324 DR examinations, follow-up imaging according to our parameters was available for 214 patients (67%). Lung nodules were found and subsequently confirmed in 35 patients (10%) without CAD. Using CAD, nodules were found and subsequently confirmed in 51 patients (15%), improving sensitivity from 63.8% (95% confidence interval [CI], 0.49%-0.76%) to 92.7% (95% CI, 0.82%-0.98%) (P < .0001, McNemar). Nodules were subsequently proved to be malignant in five of the 16 additional cases (31%). False-positive readings increased from three to six cases; specificity decreased from 98.1% (95% CI, 0.95%-0.99%) to 96.2% (95% CI, 0.92%-0.98%) (not significant). There were 153 true-negative cases (71.4%). CONCLUSIONS: This study suggests that the interpretation of chest radiographs for lung nodules can be improved using an automated CAD nodule detection system. This improvement in reader performance comes with a minimal number of false-positive interpretations. PMID- 18423314 TI - MR colonography with fecal tagging: barium vs. barium ferumoxsil. AB - RATIONALE AND OBJECTIVES: Both magnetic resonance (MR) and computed tomographic (CT) colonography are useful for colon examination. With sensitivities close to those for conventional colonoscopy (CC) for polyps, colonography has been proposed as an alternative to diagnostic CC. MR colonography (MRC) with fecal tagging may be a method of gaining further patient acceptance and widespread use, but the method has to be optimized. The aim of our study was to evaluate the quality of a new contrast agent mixture and to validate a new method for evaluating the tagging efficiency of contrast agents. MATERIALS AND METHODS: Twenty patients referred to CC underwent dark lumen MRC prior to the colonoscopy. Two groups of patients received two different oral contrast agents (barium sulfate and barium sulfate/ferumoxsil) as a laxative-free fecal tagging prior to the MRC. After MRC, the contrast agent was rated qualitatively (with the standard method using contrast-to-wall ratio) and subjectively (using a visual analog scale [VAS]) by three different blinded observers. RESULTS: Evaluated both qualitatively and subjectively, the tagging efficiency of barium sulfate/ferumoxsil was significantly better (P < .05) than barium sulfate alone. The VAS method for evaluating the tagging efficiency of contrast agents showed a high correlation (observer II, r = 0.91) to the standard method using contrast-to wall ratio and also a high interclass correlation (observer II and III = 0.89/0.85). MRC found 1 of 22 (5%) polyps <6 mm, 2 of 3 (67%) polyps 6-10 mm, and 2 of 2 (100%) polyps >10 mm. CONCLUSION: MRC with fecal tagging using barium sulfate/ferumoxsil as contrast agent will give better overall assessment of the colon wall compared to barium sulfate alone. Furthermore, the VAS method of evaluating fecal tagging efficiency correlated with the standard method of calculating the contrast-to-wall ratio. PMID- 18423315 TI - Assessing diagnostic confidence: a comparative review of analytical methods. AB - RATIONALE AND OBJECTIVES: The ability of a test to influence diagnostic confidence is used as a measure of its efficacy. Our aim was to compare analytic methods that evaluate changes in confidence. MATERIALS AND METHODS: The approaches compared were "basic," "retained diagnoses," "Omary," "Tsushima," and "score-based" methods. For illustration, data from a clinical study assessing changes in diagnostic confidence (0%-100%) before and after abdominopelvic computed tomography (CT) in patients with acute abdominal pain were used. RESULTS: The basic, retained diagnoses and Omary methods all ignore whether the test yields a correct diagnosis (confident, but incorrect, diagnoses are regarded positively). Although the Tsushima method takes some account of diagnostic accuracy, all misdiagnoses are considered equal. The score-based method addresses some of the fundamental limitations in the other analytical methods, such as diagnostic accuracy and the varying nature of different misdiagnoses. In the case study, mean (SD) diagnostic confidence for the cohort as a whole (n = 62) increased following CT: 50.7% (20.8%) to 73.2% (20.9%). Pretest diagnoses were changed following CT in 43% (27 of 62) of patients. Pretest diagnoses proved to be incorrect in 52% (32 of 62), and post-test diagnoses incorrect in as many as 19% (12 of 62) of patients. All five analytic methods indicated a positive contribution for CT (all P < or = .003). CONCLUSION: Although our illustrative case study revealed no consequential differences across the five methods, there remain substantial differences in the fundamental principles underlying them that should affect choice of analytic method when assessing diagnostic confidence. PMID- 18423316 TI - Diffusion-weighted MR: therapeutic evaluation after chemoembolization of VX-2 carcinoma implanted in rabbit liver. AB - RATIONALE AND OBJECTIVES: The study goal was to evaluate the ability of diffusion weighted imaging (DWI) in assessing the viability of rabbit liver VX-2 tumor after transcatheter arterial chemoembolization (TACE). MATERIALS AND METHODS: VX 2 tumors were grown in the livers of 19 rabbits, and chemoembolization was performed. MR imaging was acquired 1 week after TACE. The rabbits were killed for histologic investigation immediately after MR imaging, and the proportion of viable tumor was calculated based on histopathologic examination. Apparent diffusion coefficient (ADC) values were measured in viable and necrotic tumor portion, and were compared using the paired Student's t test. RESULTS: Viable tumors were absent (n = 3), less than 5% (n = 6), and 5% or more (n = 10) at pathology examination. On DWI, three tumors with no viable portion were interpreted as having no viable portion, but three of six tumors with a viable portion of less than 5% were considered as having no viable portion. The mean ADC values of necrotic and viable tumor were 1.653 +/- 0.126 mm(2)/sec and 0.883 +/- 0.407 mm(2)/sec (b = 1000 sec/mm(2)), respectively, and the ADC values of necrotic tumors were significantly greater than those in viable tumors (p < .01). CONCLUSION: Although DWI is a useful tool for assessing tumor viability, viable tumor may not be detected on DWI when it is too small. PMID- 18423317 TI - Comparison of secretin-stimulated magnetic resonance pancreatography and manometry results in patients with suspected sphincter of oddi dysfunction. AB - RATIONALE AND OBJECTIVES: To measure main pancreatic duct diameter (PDD) with magnetic resonance pancreatography (MRP) before and after secretin injection in patients with suspected sphincter of Oddi dysfunction (SOD) and to determine if the diameter change is predictive of sphincter of Oddi manometry (SOM) results. MATERIALS AND METHODS: We identified all patients during the study period referred for SOM for clinically suspected SOD; patients with an intact sphincter and without contraindication to MRP examination were considered for study entry. Consenting patients underwent MRP, including dynamic imaging of the pancreatic duct after intravenous administration of porcine secretin followed by SOM during endoscopic retrograde cholangiopancreatography. MRP was defined as abnormal when PDD remained increased by > or = 1.0 mm from baseline 15 minutes after secretin injection. SOM was abnormal when basal sphincter pressure (SP) was > or = 40 mm Hg. Mean PDD before and after secretin administration was compared within normal and abnormal SP groups with two-tailed unpaired t-test; the mean difference between baseline and peak PDD and duration of > or = 0.5 mm increase in PDD was compared between groups with two-tailed t-test. P < .05 was considered significant. RESULTS: Of 70 patients referred for SOM, 30 met all entry criteria, gave consent to participate, and underwent both MRP and SOM. Ten of 30 patients (33%) had normal SP; 20 (67%) were abnormal. PDD increased significantly after secretin injection (normal SP, 1.62 +/- 0.73 to 2.78 +/- 0.77 mm, P < .01; abnormal SP, 1.45 +/- 0.26 to 2.32 +/- 0.75 mm, P < .01). There was no difference between normal and abnormal SP groups in amount of PDD increase (1.15 +/- 0.75 vs. 0.88 +/- 0.72 mm; P = .33) or duration of > or = 0.5 mm increase in PDD (5.28 +/- 8.76 vs. 13.60 +/- 13.00 minutes; P = 0.07). CONCLUSIONS: In patients with suspected sphincter of Oddi dysfunction, magnetic resonance pancreatography demonstrated PDD increase following secretin injection but did not predict the results of manometry. PMID- 18423318 TI - Complicated breast cysts on sonography: is aspiration necessary to exclude malignancy? AB - RATIONALE AND OBJECTIVES: To determine the incidence of breast cancer in women presenting for fine needle aspiration of sonographically diagnosed complicated breast cysts. MATERIALS AND METHODS: Institutional review board approval was obtained. A retrospective study was performed of 186 consecutive women who presented for fine needle aspiration of 243 sonographic complicated cysts detected by clinical examination or imaging between January 2002 and August 2003. Sonographic complicated cysts were defined as those meeting most but not all criteria for simple cysts similar to current Breast Imaging Reporting and Data System classification. We excluded solid masses, cysts with solid components, intracystic masses, and simple cysts. Prospective mammography, ultrasound, and procedure reports were reviewed. A case was considered positive if cytology, core needle biopsy, or surgical excision was positive for cancer. Final diagnosis was established by biopsy, cytology, clinical resolution, or stability over time. RESULTS: One of 243 (0.4%) lesions proved malignant (95% confidence interval 0 1.94%); 210/243 (86.4%) of cases yielded fluid on aspiration. Of 141 samples submitted to cytology, 138 (97.9%) were benign and 3 (2.1%) atypical. All cases of atypia were benign at surgical excision. Ninety five of 243 (39.1%) aspiration samples with typical cyst fluid were discarded. A total of 33/243 (13.6%) cases did not yield fluid, 1 of which was positive for cancer. Thirty cases underwent core needle or excisional biopsy for imaging discordance with benign results. CONCLUSIONS: Breast cancer presenting as a complicated cyst by ultrasound was rare (0.4%, 95% confidence interval 0-1.94%). These results provide support for classification of complicated cysts as probably benign. PMID- 18423319 TI - Indeterminate or suspicious breast lesions detected initially with MR imaging: value of MRI-directed breast ultrasound. AB - RATIONALE AND OBJECTIVES: To retrospectively determine the value of magnetic resonance imaging (MRI)-directed breast ultrasonography (US) in the evaluation of indeterminate or suspicious lesions identified on contrast-enhanced, breast MRI. MATERIALS AND METHODS: A total of 395 patients presenting for breast MRI during a 4-year period was retrospectively reviewed. Seventy-one patients were recommended for MRI-directed US for further characterization of indeterminate or suspicious breast lesions detected on MRI. Fifty-five patients (all female; age 31-80 years) had US. Their MRI and US were reviewed and tested for correlations with histologic results or long term follow-up. Logistic regression analyses were used to test for associations between MRI lesion characteristics and US detection rate. RESULTS: US identified 46 of 97 (47%) lesions depicted at MRI from 55 patients (47 [85%] of these patients had histories of breast malignancies). Twelve cancers were found from the 97 lesions (12%). Biopsy was avoidable in 10 lesions (10%). The detection rate with US was slightly higher with "mass" (55% [23/42]) lesions described in MRI than "non-mass" lesions or lymph nodes (42% [23/55]). There was a significant positive association (odd ratio = 1.23: 95% CI = 1.05-1.43, P = .01) between US detection rate and MRI mass lesion size. There was no statistical significance between US detection rate and the presence of malignancies; 42% (5/12) of MRI malignant lesions were not visualized with US. CONCLUSIONS: MRI-directed US reduced the number of biopsies required for indeterminate or suspicious MRI lesions. Nevertheless, the lesions which were biopsied had a low rate of malignancy. PMID- 18423321 TI - 3D TOF MRA of intracranial aneurysms at 1.5 T and 3 T: influence of matrix, parallel imaging, and acquisition time on image quality - a vascular phantom study. AB - RATIONALE AND OBJECTIVES: A 3-T magnetic resonance imaging system provides a better signal-to-noise ratio and inflow effect than 1.5 T in three-dimensional time-of-flight (3D TOF) magnetic resonance angiography (MRA). The purpose of this study is to analyze the influence of matrix, parallel imaging, and acquisition time on image quality of 3D TOF MRA at 1.5 and 3 T, and to illustrate whether the combination of larger matrixes with parallel imaging technique is feasible, by evaluating the visualization of simulated intracranial aneurysms and aneurysmal blebs using a vascular phantom with pulsatile flow. MATERIALS AND METHODS: An anthropomorphic vascular phantom was designed to simulate the various intracranial aneurysms with aneurysmal bleb. The vascular phantom was connected to an electromagnetic flow pump with pulsatile flow, and we obtained 1.5- and 3-T MRAs altering the parameters of 3D TOF sequences, including acquisition time. Two radiologists evaluated the depiction of simulated aneurysms and aneurysmal blebs. RESULTS: The aneurysmal blebs were not sufficiently visualized on the high spatial resolution 1.5-T MRA (matrix size of 384 x 256 or 512 x 256), even with longer acquisition time (9 or 18 min). At 3 T with acquisition time of 4.5 min using parallel imaging technique, however, the depiction of aneurysmal blebs was significantly better for the high-spatial resolution sequence than for the standard resolution sequence. For the high-spatial resolution sequence, the longer acquisition times did not improve the depiction of aneurysmal blebs in comparison with 4.5 min at 3 T. CONCLUSIONS: For 3D TOF MRA, the combination of the large matrix with parallel imaging technique is feasible at 3 T, but not at 1.5 T. PMID- 18423320 TI - Evaluating the effect of image preprocessing on an information-theoretic CAD system in mammography. AB - RATIONALE AND OBJECTIVES: In our earlier studies, we reported an evidence-based computer-assisted decision (CAD) system for location-specific interrogation of mammograms. A content-based image retrieval framework with information theoretic (IT) similarity measures serves as the foundation for this system. Specifically, the normalized mutual information (NMI) was shown to be the most effective similarity measure for reduction of false-positive marks generated by other prescreening mass detection schemes. The objective of this work was to investigate the importance of image filtering as a possible preprocessing step in our IT-CAD system. MATERIALS AND METHODS: Different filters were applied, each one aiming to compensate for known limitations of the NMI similarity measure. The study was based on a region-of-interest database that included true masses and false-positive regions from digitized mammograms. RESULTS: Receiver-operating characteristics (ROC) analysis showed that IT-CAD is affected slightly by image filtering. Modest, yet statistically significant, performance gain was observed with median filtering (overall ROC area index A(z) improved from 0.78 to 0.82). However, Gabor filtering improved performance for the high-sensitivity portion of the ROC curve where a typical false-positive reduction scheme should operate (partial ROC area index (0.90)A(z) improved from 0.33 to 0.37). Fusion of IT-CAD decisions from different filtering schemes markedly improved performance (A(z) = 0.90 and (0.90)A(z) = 0.55). At 95% sensitivity, the system's specificity improved by 36.6%. CONCLUSIONS: Additional improvement in false-positive reduction can be achieved by incorporating image filtering as a preprocessing step in our IT-CAD system. PMID- 18423322 TI - Comparison between malignant and benign abdominal lymph nodes on diffusion weighted imaging. AB - RATIONALE AND OBJECTIVES: The purpose of this study is to review the apparent diffusion coefficient (ADC) values of benign and metastatic abdominal lymph nodes on diffusion-weighted imaging (DWI). MATERIALS AND METHODS: Twenty-eight patients with a total of 40 benign (20 patients) and 16 malignant (8 patients) lymph nodes who underwent DWI MRI of the abdomen (b = 0.600) were enrolled in the study. ADC values of the lymph nodes were measured and comparison was made between benign and malignant groups. RESULTS: Mean ADC value of lymph nodes was 2.38 +/- 0.29 and 1.84 +/- 0.37 x 10(-3) mm(2)/sec in the benign and malignant groups, respectively. There was a significant statistical difference between the ADC values of benign and malignant lymph nodes (P < .0005). CONCLUSION: A wide range of ADC values exist in patients with metastatic abdominal lymph nodes, with a tendency of higher ADC values in benign lymph nodes. PMID- 18423325 TI - Names make news. PMID- 18423324 TI - Critique of the transitional year internship and its relationship to radiology residency. AB - RATIONALE AND OBJECTIVES: The purpose of the study is to determine if transitional year program (TYP) requirements foster realization of standards of excellence and clinical relevance for future radiologists and to explore demographic and economic factors pertinent to TYPs. MATERIALS AND METHODS: A list of accredited TYPs were obtained from the American Medical Association's Graduate Medical Education (ACGME) Directory 2006-2007. Specialty distribution of TYP graduates was examined from statistics provided by the ACGME, and data from the 2007 Main Residency Match was analyzed. Data derived from a concurrent survey of the perception of the value of internship sent to all current radiology residents and fellows was assessed. The institutional costs of employing TYP interns versus physician assistants were also calculated. RESULTS: Forty-one of the 125 TYPs lack residencies in internal medicine (IM), general surgery (GS), or both, and approximately two-third of these lack full medical school affiliation. The interns who will graduate from these 41 programs account for 103 of the 1,128 radiology residents in their post-graduate year 2. Despite the longest elective time offered in TYPs compared to conventional preliminary programs, current radiology trainees who had participated in preliminary IM or GS internships were more satisfied compared to trainees completing TYPs. CONCLUSIONS: The requirements of the transitional internship and compliance with them need to be carefully assessed to determine their efficacy. Despite the strong economic impetus for hiring TYP interns, the availability of open slots in existing preliminary programs in IM and GS, coupled with radiology residents' greater level of satisfaction with traditional over transitional internships, makes the existence of TYPs less compelling. PMID- 18423323 TI - Recent developments in the Dorfman-Berbaum-Metz procedure for multireader ROC study analysis. AB - RATIONALE AND OBJECTIVES: The Dorfman-Berbaum-Metz (DBM) method has been one of the most popular methods for analyzing multireader receiver-operating characteristic (ROC) studies since it was proposed in 1992. Despite its popularity, the original procedure has several drawbacks: it is limited to jackknife accuracy estimates, it is substantially conservative, and it is not based on a satisfactory conceptual or theoretical model. Recently, solutions to these problems have been presented in three papers. Our purpose is to summarize and provide an overview of these recent developments. MATERIALS AND METHODS: We present and discuss the recently proposed solutions for the various drawbacks of the original DBM method. RESULTS: We compare the solutions in a simulation study and find that they result in improved performance for the DBM procedure. We also compare the solutions using two real data studies and find that the modified DBM procedure that incorporates these solutions yields more significant results and clearer interpretations of the variance component parameters than the original DBM procedure. CONCLUSIONS: We recommend using the modified DBM procedure that incorporates the recent developments. PMID- 18423326 TI - Bad news. PMID- 18423327 TI - Contact lens & anterior eye. Editorial. PMID- 18423328 TI - Differences in effective connectivity between dyslexic children and normal readers during a pseudoword reading task: an fMRI study. AB - PURPOSE: This fMRI study investigated phonological and lexicosemantic processing in dyslexic and in chronological age- and reading level-matched children in a pseudoword reading task. MATERIALS AND METHODS: The effective connectivity network was compared between the three groups using a structural model including the supramarginal cortex (BA 40; BA: Brodmann area), fusiform cortex (BA 37) and inferior frontal cortex (BA 44/45) areas of the left hemisphere. RESULTS: The results revealed differences in connectivity patterns. In dyslexic patients, in contrast with chronological age- and reading level-matched groups, no causal relationship was demonstrated between BA 40 and BA 44/45. However, a significant causal relationship was demonstrated between BA 37 and BA 44/45 both in dyslexic children and in the reading level-matched group. CONCLUSIONS: These findings were interpreted as evidence for a phonological deficit in developmental dyslexia. PMID- 18423330 TI - Thymocognitive input and postural regulation: a study on obsessive-compulsive disorder patients. AB - CONTEXT: To show that emotional and cognitive information acts upon the postural balance system in a way comparable to that of the other known inputs (vision, vestibular, proprioception). METHOD: Controlled case study on 90 subjects. One group was composed of 45 subjects suffering from obsessive-compulsive disorder (OCD) in accordance with the Yale-Brown scale, while the other was the control group. All of the subjects underwent recording of their orthostatic posture on a force platform with eyes open and eyes closed. RESULTS: As regards to the postural findings, the two groups appear to be quite different. The OCD patients present a considerably reduced area and velocity of sway regardless of whether their eyes are open or closed. CONCLUSION: These results are coherent with regard to those of other studies establishing the link between postural balance and psychological status. Recent morphological studies likewise tend to confirm the existence of neuronal networks common to postural regulation and cognitive and emotional functioning. When interpreting symptoms, these interactions should be taken into account. PMID- 18423329 TI - Alcoholism leads to early perceptive alterations, independently of comorbid depressed state: an ERP study. AB - INTRODUCTION: Alcoholism is associated with a deficit in the processing of emotional facial expressions (EFE) and with a delayed P3b component, partially mediated by earlier perceptive deficits (P100, N170). Since alcohol dependence often occurs with depression, we aim at investigating whether classical event related potentials (ERP) alterations observed in alcoholism are modulated or not by depression. METHODS: Four groups (controls; alcoholics; depressed; alcoholics depressed) of 12 participants performed two different discrimination tasks, a gender and an emotional one. They had to decide as quickly as possible about the gender or the emotion displayed by facial stimuli during an ERP recording session (32 channels). Reaction times (RTs), P100, N100, N170 and P3b were recorded. RESULTS: At the behavioural level, control participants discriminated EFE (but not gender) more rapidly than the three other groups. At the ERP level, the differences observed on RTs for emotional task were neurophysiologically indexed by a delayed P3b component. This delay was associated with earlier ERP alterations (P100, N100, N170), but only in participants suffering from alcohol dependence, in association or not with depression. DISCUSSION: On the one hand, individuals with alcoholism, associated or not with a comorbid depression, were impaired in the processing of EFE. This deficit was neurophysiologically indexed by early perceptive (P100, N100, N170) and decisional (P3b) alterations. On the other hand, non-alcoholic patients with depression only exhibited P3b impairment. These results lead to potential implications concerning the usefulness of the ERP for the differential diagnosis in psychiatry, notably concerning the comorbidities in alcoholism. PMID- 18423331 TI - Effect of speed on kinematic, kinetic, electromyographic and energetic reference values during treadmill walking. AB - OBJECTIVE: Evaluation of normal and pathological gait on the level ground has drawbacks that could be overcome by walking on a treadmill. The present work was designed to assess the feasibility of extended gait analysis on a treadmill allowing multiple steps recording at a constant speed in young healthy subjects. It also aimed to provide speed-specific kinematic, kinetic, electromyographic and energetic reference values. METHOD: Twelve healthy volunteers (23 +/- two years) walked on a force measuring treadmill at six speeds (1-6 k mh(-1)). Kinematics and kinetics were analysed at the hip, knee and ankle. Electromyographic muscle activity timing of quadriceps femoris, biceps femoris, tibialis anterior and lateral gastrocnemius was recorded. The energy cost was computed from oxygen consumption measurement. RESULTS: All variables were speed-dependent. Kinematics and kinetics peaks amplitude increased and occurred earlier during the walking cycle with increasing walking speed. Muscle activity timing also changed with speed, although the number of bursts remained constant. The energetic cost presented a U-shaped curve, with minimal values around 4 km h(-1). Data were compared to overground walking data obtained by several authors: all results, except kinetic ones, were similar, turning down the thought that biomechanics of treadmill and overground walking could be different. CONCLUSION: This study provides reference values for normal and pathological walking on treadmill and allows speed-dependent comparison between subjects. PMID- 18423332 TI - [Evolution of locomotion disorders in Huntington's disease]. AB - INTRODUCTION: Locomotion disorders are important in Huntington's disease (HD). Although the rates of evolution of motor, functional or cognitive aspects of HD have been studied, the evolution of locomotion disorders in early stages of the disease remains unknown. OBJECTIVES: To determine the rate of evolution of the HD associated gait and gait initiation disorders and their correlates. PATIENTS AND METHODS: Eighteen HD patients were recorded with a minimum interevaluation interval of one year. Akinesia was studied by evaluating the anticipatory postural adjustment (APA) phase preceding the first step. We also evaluated gait speed, stride time and stride length. RESULTS: We observed an alteration in the APA phase, whose evolution was correlated with that of akinesia. We also observed a decrease in gait speed, which was due both to an increase in stride time and a decrease in stride length. Stride-to-stride variability did not worsen between both evaluations. CONCLUSIONS: A worsening in both gait initiation and gait performance was observed in HD. Initial weak functional capacity and more severe motor impairment seem to be associated with a faster progression of locomotion parameters in these mildly impaired HD patients. PMID- 18423333 TI - Comparison between nerve conduction studies and current perception threshold test in carpal tunnel syndrome. AB - AIM OF STUDY: Nerve conduction studies (NCS) only test large myelinated A(alpha) or A(beta) nerve fibers, whereas the current perception threshold (CPT) test has been suggested to evaluate a wide range of nerve fibers (A(beta), A(delta) and C). This study was undertaken to compare CPT and the standard NCS test by Bland's severity scale with the patient-based measurement of symptoms and functional status of the hand by Boston CTS questionnaire assessment. PATIENTS AND METHODS: We performed NCS and CPT on 31 patients (mean age 54.6+/-11.7 years; 31-79 years) with clinical diagnosis of CTS. NCS severity was classified according to Bland's scale and CPT was measured at 2000, 250 and 5 Hz and severity was graded between 0 and 12. Two-tailed Spearman's correlation analysis was performed to assess correlations between Boston questionnaire score and Bland's severity scale and CPT total score. RESULTS: The results showed that Bland's scale, based on NCS, had more significant correlations with symptoms (Spearman's rho=0.402, p=0.002) and function (rho=0.400, p=0.001) than CPT total scores (rho=0.200, p=0.135; rho=0.234, p=0.069). In CPT, only score measure at 2000 Hz showed a significant correlation with Boston CTS questionnaire scores (with symptom rho=0.308, p=0.020; with function rho=0.302, p=0.018), whereas those measured at 250 Hz and 5 Hz did not (p>0.05). CONCLUSION: Though CPT may have a supplementary role in the diagnosis of CTS, NCS better reflects patients' symptoms and functions than CPT on the patient's perspective. PMID- 18423334 TI - Accessory nerve stimulation: motor response of the sternocleidomastoid muscle. AB - AIMS: The aim of this study is to describe a method for recording compound muscle action potential (CMAP) from the sternocleidomastoid (SCM) in response to accessory-nerve stimulation. METHODS: Fifty-four subjects were included. Accessory nerve was stimulated at neck in two different positions; CMAPs were recorded with surface electrodes. Motor amplitude, area, duration, distal latency and intensity of stimulation at different sites were evaluated. RESULTS: CMAPs were evoked in every subject. We did not observe interside differences or variations related to age. Stimulation behind SCM evoked CMAPs with slightly shorter latencies and with less stimulus intensity. CONCLUSIONS: CMAPs of the SCM can be easily obtained by accessory nerve stimulation at the neck; this method allows a better determination of the accessory nerve lesion site. In addition, this method can be useful in the evaluation of patients with amyotrophic lateral sclerosis. PMID- 18423336 TI - An additional site of pudendal nerve compression? PMID- 18423335 TI - Quantitative EEG and cognitive evoked potentials in anemia. AB - OBJECTIVE: The anemic status may alter brain functions and electrogenesis, as reflected by EEG and cognitive EPs (CEPs). This study aims to evaluate CEPs and EEG power spectra in adult patients with iron-deficiency anemia and to determine the effects of appropriate iron therapy on electrodiagnostic findings. METHODS: Fifty-one patients with iron-deficiency anemia underwent CEP and EEG recording. All patients were re-assessed after three months of oral-iron therapy. RESULTS: All patients had recovered from their anemia through the three-month iron therapy. Central N1 amplitude and parietal P2 amplitude was increased. N2 latencies were shortened in frontal and central regions. P3 latencies were shortened in frontal, central and parietal areas and P3 amplitude was increased in the parietal region. Except in the gamma-band, all pretreatment and post treatment mean-power values were significantly lower at the temporal, parietal and occipital regions. CONCLUSIONS: This study indicates that in iron-deficiency anemia, appropriate iron therapy can improve brain electrogenesis, as reflected by P300 and EEG power spectra. PMID- 18423338 TI - Caution cutting the canthus: thoughts on microphthalmic expansion. PMID- 18423339 TI - Of Pax2 laboratory mice and human papillorenal investigations: maintaining the distinctions between cause and effect. PMID- 18423340 TI - Motor development and surgery for infantile esotropia. PMID- 18423341 TI - Effect of axial length and keratometry measurement error on intraocular lens implant power prediction formulas in pediatric patients. AB - PURPOSE: To examine the relationship between axial length and keratometry measurement errors and intraocular lens (IOL) power calculations for pediatric eyes. METHODS: The sensitivity of IOL power calculation to errors in axial length and keratometry measurements was computed as a function of axial length and keratometry for the SRK II, Hoffer Q, Holladay I, SRK/T, and Haigis formulas. RESULTS: The sensitivity of the IOL power calculation to an axial length measurement error is increased at 4 to 14 D/mm error in axial length in children compared with 3 to 4 D/mm error in axial length in adults. The error in calculation is 0.8 to 1.3 D/D error in keratometry measurement for both children and adults. CONCLUSIONS: Axial length measurement errors in pediatric eyes may lead to large errors in IOL power calculations. PMID- 18423342 TI - Bilateral vitreous hemorrhage in a child due to isolated platelet factor 3 availability defect. AB - Vitreous hemorrhage is an uncommon cause of decreased vision in children. We report a 3-year-old child with bilateral vitreous hemorrhage secondary to an isolated platelet factor 3 (PF-3) availability defect. Prophylactic infusion of platelet-rich plasma followed by pars plana vitrectomy in both eyes resulted in an excellent surgical and visual outcome. This report emphasizes the importance of exhaustive hematological and platelet function studies in a child presenting with bilateral vitreous hemorrhage and its implications on the management of present and future bleeding episodes. PMID- 18423343 TI - A new conformer design to expand the horizontal lid fissure in severely microphthalmic eyes. AB - Effective treatment of severe microphthalmos requires early placement of conformers to increase the size of the conjunctival sac and to increase orbital dimensions. This process may be confounded by inadequate horizontal lid length and microblepharon. Lateral canthotomy can be used to facilitate early placement of serial conformers, but this procedure is often avoided due to frequent peripalpebral scarring and recurrent phimosis. Conformers currently in use do not separate the lateral lid margins enough to prevent such scarring after lateral canthotomy. We describe a child with severe microphthalmos and microblepharon in whom a novel conformer was used to prevent scarring following lateral canthotomy. Use of this conformer design may make lateral canthotomy a viable option to expand the horizontal lid fissure and facilitate early orbital expansion in patients with severe microphthalmos. PMID- 18423345 TI - Long-term follow-up of visual functions in prematurely born children--a prospective population-based study up to 10 years of age. PMID- 18423347 TI - Team care of the patient with cleft and lip palate. Foreword. PMID- 18423348 TI - Team care of the patient with cleft lip and palate. PMID- 18423350 TI - Marfan syndrome and sudden death within a family - aetiologic, molecular and diagnostic issues at autopsy. AB - Although Marfan syndrome has a range of characteristic morphological features involving the ocular, cardiovascular and musculoskeletal systems, the phenotype is variable. In addition, mutations have been identified in the gene encoding for fibrillin-1 and also in the transforming growth factor-beta receptor 2 (TGF betaR2) gene. Two cases are presented of sudden and unexpected deaths in cousins who manifested morphologic features of Marfan syndrome at autopsy. Case 1: A 36 year-old male who collapsed and was found at autopsy to have arachnodactyly, a high arched palate and lethal aortic dissection with haemopericardium. Case 2: A 34-year-old male who collapsed and was found at autopsy to have arachnodactyly, a high arched palate, pes cavus and a dysplastic mitral valve. Current aetiological theories and molecular findings are discussed. While family follow-up and counselling are advised when cases come to autopsy, given the variability in phenotype and genotype, and the difficulties that exist in attempting to determine clinical prognosis from either of these, such deaths may raise more concerns for surviving family members than providing answers. PMID- 18423351 TI - Photography, patient consent and scientific publications: medicolegal aspects in France. AB - To take a photograph of a person is to lay bare their identity to the eyes of others. The photograph generates an ambiguous relationship with the idea of identity. It can in turn lay it bare, exploit it, reveal and embody it. It creates an image which takes on its own existence separately from the person portrayed. It can become a source of profit, a cause of moral harm, a means of proof, and an object of cupidity. The question which arises is: how can we know when it is legitimate to use a photograph? The law protects the person's legitimate interest, but does not only protect private interests, it also watches over the common good, and the interest of society as a whole justifies a certain number of uses of a person's photograph without their consent. This article has been written in order to clarify the ethical and legal conflicts from a French perspective, which the physician has to confront when obtaining consent from a patient before taking a photograph. Awareness of these points should make it possible to avoid any problems which could arise in relation to publications which carry illustrations for the purpose of increasing their impact. PMID- 18423352 TI - Are there age-related effects in antisocial personality disorders and psychopathy? AB - We investigated possible age differences in the frequency of antisocial personality disorder (ASPD), and of psychopathy according to Hare's criteria and its constituent features: In a cross-sectional study 226 male violent offenders, detained in adult or youth custody, were investigated using the Psychopathy Checklist Screening Version (PCL:SV) and the SCID II Interview. Their ages ranged from 18 to 59 years. Total PCL:SV score was negatively correlated with age. ANOVA showed that total PCL scores for three age groups differed significantly. Both effects were due entirely to Factor 2 of the PCL. Factor 1 was not related to age. The frequency of ASPD was also lower among older prisoners. The relationship with age was similar to that of PCL:SV Factor 2. These results point to age related effects in psychopathy and suggest that different aspects of psychopathy follow different developmental courses. The results of our group comparison suggest that the different subfacets of psychopathy are not stable over time to the same extent. In order to make statements about the course of intraindividual development, however, longitudinal studies would be required. PMID- 18423353 TI - Massive internal injury in the absence of significant external injury after collisions of passenger vehicles with much larger vehicles. AB - We have encountered cases of motor vehicle collisions in which there was relatively little external trauma, but there were massive internal injuries that were much more extensive than might be expected from the external examination. Two cases were collisions between trains and pickup trucks, the third a collision between a semi trailer and a van. In all three cases, the external examination showed minor abrasions and lacerations. Internally, however, there were massive injuries which were fatal. While the significant injuries in our cases were not surprising given the force of the collisions involved, it is interesting that the external examination showed relatively little injury. We speculate that collisions between passenger vehicles and very large vehicles generate massive internal injuries by transmission of force through the victims. PMID- 18423354 TI - Foot index: is it a tool for sex determination? AB - Identification of an individual is of paramount importance in forensic investigations. The dimensions of the foot can be used for the determination of sex and stature of an individual in forensic investigations. No systematic studies are available on the determination of sex from foot measurements of North Indians. Therefore, foot index is derived to determine the sex of an individual in a single community of North India. The foot index for both genders is derived by dividing the foot breadth by foot length and multiplying it by hundred. In the present investigation, the foot index is found to be slightly higher in females in the right foot and males in the left foot. The study suggests that although foot length and foot breadth show significant sex differences, sex determination cannot be made conclusively from the foot index. PMID- 18423355 TI - Police custody deaths in Maryland, USA: an examination of 45 cases. AB - Previous published research suggests there are a diverse, yet relatively consistent, set of factors present in sudden, unexpected, and initially inexplicable deaths in police custody. This retrospective analysis examines police custodial deaths in Maryland, USA. Police custody death is operationalized to include deaths which occurred suddenly and unexpectedly during police/citizen encounters. Only deaths for which the cause was initially undeterminable are included. Suicidal hanging, police shootings and fatal pursuits are not included. Review of the records at Maryland's Office of the Chief Medical Examiner indicates that 45 persons died suddenly and unexpectedly in the custody of the police, between 1990 and 2004. Commensurate with existing research, our analysis identifies multiple factors generally present in this type of police custody death. Although individual-specific, these factors include a relatively static constellation of behavioral (e.g., erratic and/or violent behavior, and physical struggle) and physical (e.g., stimulant abuse, natural disease, and obesity) dimensions. While the presence of these factors has been well-documented, their empirical significance, interactions and causal sequence have yet to be established. To increase the methodological rigor of subsequent research, we have developed a comprehensive, national custody death database by which multivariate models may be analyzed. PMID- 18423356 TI - Treating cocaine body packers: the unspoken personal risks. AB - Cocaine trafficking is a significant problem that many Caribbean territories must face. "Body packing" is a common method of transport where the smugglers ingest several cocaine filled packages. Body packers may be taken to hospital when they are detained by law enforcement officers, but occasionally they present on their own or accompanied by persons other than the authorities. This scenario poses a difficult management dilemma in any jurisdiction. We describe our experience with one such case in Jamaica. PMID- 18423357 TI - Safe in our hands?: a study of suicide and self-harm in asylum seekers. AB - This study examined the incidence of suicide and self-harm in asylum seekers in the UK, both those in detention and in the community. The investigation revealed that data recording is seriously flawed or sometimes non-existent. However, the scanty data those were available from Immigration Removal Centres, coroners' records and Prison Ombudsman's reports showed high levels of self-harm and suicide for detained asylum seekers as compared with the United Kingdom prison population. It is suggested that this could be attributed to routine failure to observe and mitigate risk factors. The author makes the following recommendations: coroners should record asylum seeker status and ethnicity of deceased, self-harm monitoring in the community should record asylum seeker status and ethnicity, health care in immigration removal centres should meet the same standards as UK prisons as a minimum, allegation of torture by immigration detainees should trigger a case management review and risk assessment for continued detention, and this process should be open to audit, and interpreters should be used for mental state examinations unless their English has been shown to the fluent. PMID- 18423358 TI - Death certification practices of forensic physicians within the Strathclyde region of Scotland, UK. AB - Data relating to the perceived and actual cause of death, in those dying in the community is sparse. This in part is related to how each country deals with death investigations and certification of death. Under common law in Scotland, sudden unexpected deaths are referred to the Procurator Fiscal. Autopsy is not always the final arbiter of truth. While actual figures are not available at present, anecdotally the police indicate that general practitioners (GPs) are often declining to attend or certify in the vast majority of cases. The purpose of this prospective observational study was to examine the certification practice of the forensic physicians working for Strathclyde Police. In conclusion, the results from this short study indicate that in cases where the cause of death for an individual, who died suddenly in the community and where an autopsy is deemed necessary, forensic physicians are no better or worse than a pathologist in predicting the underlying reason. PMID- 18423359 TI - A unique trend of murder-suicide in the Jamnagar region of Gujarat, India (a retrospective study of 5 years). AB - Jamnagar region, Gujarat state, enjoys a relatively low incidence of homicide in India. In the 5 year period from 2000 to 2004, 8 mothers committed 13 murders involving 3 male and 10 female victims and in every case it was followed by suicide of the assailant mothers. During the study the annual incidence of murder suicide was about 1.8 cases. All the assailants were mothers and the victims were their small children in the age group of 6 months to 7 years. Five incidents took place in rural areas and three in urban areas. It was prevalent only in low socio economic families. Methods both for killing and suicide were either burning or drowning. All the mothers were legally married and living with the family. Family and family related matters were the main motives for killing. In one case there was history of depression of the mother due to her previous miscarriage. Alcohol consumption or drug abuse was not seen even in a single case. All cases fell in the altruistic category of filicide-suicide. PMID- 18423360 TI - Prolonged cholinergic crisis and compartment syndrome following subcutaneous injection of an organophosphate compound for suicide attempt. AB - A case of poisoning with highly lipid soluble organophosphate compound, fenthion is reported in which cholinergic crisis recurred upto 25 days following a suicide attempt. Subcutaneous injection of fenthion in the antecubital fossa by the patient produced massive swelling, cellulitis and compartment syndrome of the left arm. Emergency fasciotomy helped in restoration of circulation and saved the limb from being amputated. PMID- 18423361 TI - A battered child case with duodenal perforation. AB - Battered child syndrome can refer to children exposed to harmful, non-accidental and preventable physical treatment by those are responsible for their care which prevents the child's physical, cognitive and spiritual development. A 28 months old boy was submitted to hospital due to abdominal blunt trauma. He had been firstly applied to Isparta Children Hospital by his parents with the complaint of fever. In the first examination, he was conscious, his general condition was poor there was respiratory acidosis, and neck stiffness was present. There were several fresh traumatic lesions on his face and left arm. His complaints were thought due to meningitis and antibiotics were started. He was transported to Suleyman Demirel University Hospital after a day because of vomiting, abdominal pain, tender, distended and silent abdomen, and air-fluid levels in direct abdominal X-rays. An old fracture of the right 9th rib was detected with chest X ray in university hospital. Additionally, abdominal ultrasound scan showed distended bowel loops filled with fluid. Laparotomy revealed a complete rupture of the junction of the third and fourth parts of the duodenum and several hemorrhagic regions on bowel loops. The patient was discharged after 42 days. This case report described the case through both medical and legal processed in Turkey. PMID- 18423362 TI - Accidental ligature strangulation due to crop thrasher. AB - Forty five years male was accidentally strangled when his shawl was caught in a moving electrical crop thrasher used for separating the grains from dry crops kept after cutting in the field. He was immediately taken to the nearest hospital. He was unconscious. He died after six days. Accidental ligature strangulation in this fashion due to such electrical machine is very rare and not reported previously to the best of our knowledge. PMID- 18423363 TI - Post-traumatic lethal carotid-cavernous fistula. AB - The authors report about an unexpected death by traumatic lesion of the internal carotid artery in a 30-year-old man who had fallen two metres. The man suffered a fracture of the left maxillary sinus and a fracture of the right orbit with bilateral haemorrhage of the maxillary sinus. Surgical treatment was performed with favorable outcome. Clinically, there were no neurological symptoms but about 60 days after his accident, the man died from uncontrolled epistaxis. He was submitted to the autopsy that show a linear fracture in the left side of the turcic sella and lesion of the left internal carotid artery with carotid cavernous fistula. PMID- 18423364 TI - Pericial analysis of a dental element found inside food used for human consumption. AB - The field of forensic dentistry embodies the human identification in a lot of conditions. A case is described when a tooth-like object was found in a pork sausage. A lawsuit was filed against the food company and the possible dental element was analysed. The tests used to analyze the case were: scanning electronic microscopic exam, radiographic exam and compared anatomy. The results confirmed that the object found in the sausage pack was a dental element from an adult pig; which had the potential for problems like asphyxia or contamination. The case illustrates one facet of forensic dentistry in Brazil. PMID- 18423366 TI - Tyramine oxidation by copper/TPQ amine oxidase and peroxidase from Euphorbia characias latex. AB - Tyramine, an important plant intermediate, was found to be a substrate for two proteins, a copper amine oxidase and a peroxidase from Euphorbia characias latex. The oxidation of tyramine took place by two different mechanisms: oxidative deamination to p-hydroxyphenylacetaldehyde by the amine oxidase and formation of di-tyramine by the peroxidase. The di-tyramine was further oxidized at the two amino groups by the amino oxidase, whereas p-hydroxyphenylacetaldehyde was transformed to di-p-hydroxyphenylacetaldehyde by the peroxidase. Data obtained in this study indicate a new interesting scenario in the metabolism of tyramine. PMID- 18423367 TI - p-dimethylaminocinnamaldehyde derivatization for colorimetric detection and HPLC UV/vis-MS/MS identification of indoles. AB - Cytochrome P450 2A13 (CYP2A13) is a lung specific enzyme known to activate the potent tobacco procarcinogen 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) into two carcinogenic metabolites. CYP2A13 has been crystallized and X-ray diffraction experiments illuminated the structure of this enzyme, but with an unknown ligand present in the enzyme active site. This unknown ligand was suspected to be indole but a selective method had to be developed to differentiate among indole and its metabolites in the protein sample. We successfully modified a microbiological colorimetric assay to spectrophotometrically differentiate between indole and a number of possible indole metabolites in nanomolar concentrations by derivatization with p dimethylaminocinnamaldehyde (DMACA). Further differentiation of indoles was made by mass spectrometry (HPLC-UV/vis-MS/MS) utilizing the chromophore generated in the DMACA conjugation as a UV signature for HPLC detection. The ligand in the crystallized protein was identified as unsubstituted indole, which facilitated refinement of two alternate conformations in the CYP2A13 crystal structure active site. PMID- 18423368 TI - Cytochrome c assembly: a tale of ever increasing variation and mystery? AB - Formation of cytochromes c requires a deceptively simple post-translational modification, the formation of two thioether bonds (or rarely one) between the thiol groups of two cysteine residues found in a CXXCH motif (with some occasional variations) and the vinyl groups of heme. There are three partially characterised systems for facilitating this post-translational modification; within these systems there is also variation. In addition, there are clear indications for two other distinct systems. Here some of the current issues in understanding the systems are analysed. PMID- 18423369 TI - Membrane partitioning of various delta-opioid receptor forms before and after agonist activations: the effect of cholesterol. AB - Lipid rafts depicted as densely packed and thicker membrane microdomains, based on the dynamic clustering of cholesterol and sphingolipids, may help as platforms involved in a wide variety of cellular processes. The reasons why proteins segregate into rafts are yet to be clarified. The human delta opioid receptor (hDOR) reconstituted in a model system has been characterised after ligand binding by an elongation of its transmembrane part, inducing rearrangement of its lipid microenvironment [Alves, Salamon, Hruby, and Tollin (2005) Biochemistry 44, 9168-9178]. We used hDOR to understand better the correlation between its function and its membrane microdomain localisation. A fusion protein of hDOR with the Green Fluorescent Protein (DOR*) allows precise receptor membrane quantification. Here we report that (i) a fraction of the total receptor pool requires cholesterol for binding activity, (ii) G-proteins stabilize a high affinity state conformation which does not seem modulated by cholesterol. In relation to its distribution, and (iii) a fraction of DOR* is constitutively associated with detergent-resistant membranes (DRM) characterised by an enrichment in lipids and proteins raft markers. (iv) An increase in the quantity of DOR* was observed upon agonist addition. (v) This DRM relocation is prevented by uncoupling the receptor-G-protein interaction. PMID- 18423370 TI - Electrofused giant protoplasts of Saccharomyces cerevisiae as a novel system for electrophysiological studies on membrane proteins. AB - Giant protoplasts of Saccharomyces cerevisiae of 10-35 microm in diameter were generated by multi-cell electrofusion. Thereby two different preparation strategies were evaluated with a focus on size distribution and "patchability" of electrofused protoplasts. In general, parental protoplasts were suitable for electrofusion 1-12 h after isolation. The electrophysiological properties of electrofused giant protoplasts could be analyzed by the whole-cell patch clamp technique. The area-specific membrane capacitance (0.66+/-0.07 microF/cm(2)) and conductance (23-44 microS/cm(2)) of giant protoplasts were consistent with the corresponding data for parental protoplasts. Measurements with fluorescein-filled patch pipettes allowed to exclude any internal compartmentalisation of giant protoplasts by plasma membranes, since uniform (diffusion-controlled) dye uptake was only observed in the whole-cell configuration, but not in the cell-attached formation. The homogeneous structure of giant protoplasts was further confirmed by the observation that no plasma membrane associated fluorescence was seen in the interior of giant cells after electrofusion of protoplasts expressing the light-activated cation channel Channelrhodopsin-2 (ChR2) linked to yellow fluorescent protein (YFP). Patch clamp analysis of the heterologously expressed ChR2-YFP showed typical blue light dependent, inwardly-directed currents for both electrofused giant and parental protoplasts. Most importantly, neither channel characteristics nor channel expression density was altered by electric field treatment. Summarising, multi-cell electrofusion increases considerably the absolute number of membrane proteins accessible in patch clamp experiments, thus presumably providing a convenient tool for the biophysical investigation of low signal transporters and channels. PMID- 18423371 TI - Proteins and cholesterol-rich domains. AB - Biological membranes are composed of many molecular species of lipids and proteins. These molecules do not mix ideally. In the plane of the membrane components are segregated into domains that are enriched in certain lipids and proteins. Cholesterol is a membrane lipid that is not uniformly distributed in the membrane. Proteins play an important role in determining cholesterol distribution. Certain types of protein lipidation are known to cause the lipoprotein to sequester with cholesterol and to stabilize cholesterol-rich domains. However, proteins that are excluded from such domains also contribute to the redistribution of cholesterol. One of the motifs that favor interaction with cholesterol is the CRAC motif. The role of the CRAC motif of the gp41 fusogenic protein of HIV is discussed. The distribution of the multianionic lipid, phosphatidylinositol(4,5)bis-phosphate (PtnIns(4,5)P2), is also not uniform in cell membranes. This lipid has several functions in the cell, including a morphological role in determining the sites of attachment of the actin cytoskeleton to the plasma membrane. PtnIns(4,5)P2 is sequestered by proteins having clusters of cationic residues in their sequence. Certain proteins containing cationic clusters also contain moieties such as myristoylation or a CRAC segment that would also endow them with the ability to sequester to a cholesterol-rich domain. These proteins interact with PtnIns(4,5)P2 in a cholesterol-dependent manner forming domains that are enriched in both cholesterol and in PtnIns(4,5)P2 but can also be distinct from liquid-ordered raft-like domains. PMID- 18423372 TI - Heterogeneity in binding and gene-expression regulation by HIF-2alpha. AB - Here, we demonstrate by chromatin immunoprecipitation that the binding of hypoxia inducible factors to gene regulatory regions is differentially influenced in cancer cells. Binding of HIF-2alpha varies depending on hypoxic conditions, although HIF-1alpha is constantly bound to these regions. We found by RNA interference experiments that HIF-2alpha plays a minor role in VEGF gene upregulation under hypoxia or CoCl(2) treatment, even when both HIFs are similarly bound to the promoter region. HIF-2alpha activated or suppressed the ENO1 gene under various conditions, irrespective of promoter binding. We additionally found that HIF dependence on EPO gene induction could be altered depending on the conditions, irrespective of the binding pattern of HIFs. These results demonstrate that, unlike HIF-1alpha, HIF-2alpha differentially binds and regulates transcription under hypoxia. PMID- 18423373 TI - Stimulated endocytosis in penetratin uptake: effect of arginine and lysine. AB - Cell-penetrating peptides can deliver macromolecular cargo into cells and show promise as vectors for intracellular drug delivery. Internalization occurs predominantly via endocytosis, but the exact uptake mechanisms are not fully understood. We show quantitatively how penetratin, a 16-residue cationic peptide, stimulates fluid-phase endocytosis and triggers its own uptake into Chinese hamster ovarian cells, using a 70kDa dextran to indicate macropinocytosis. The total cellular endocytotic rate is significantly less affected and we therefore propose up-regulation of macropinocytosis to occur at the expense of other types of endocytosis. By comparing penetratin to its analogs PenArg and PenLys, enriched in arginines and lysines, respectively, we show how these side-chains contribute to uptake efficiency. The degree of peptide and dextran uptake follows similar patterns regarding peptide concentration and arginine/lysine content (PenArg>penetratin>PenLys), indicating that a high content of arginines is beneficial but not necessary for stimulating endocytosis. PMID- 18423374 TI - UDP-glucuronosyltransferases 1A6 and 1A10 catalyze reduced menadione glucuronidation. AB - Menadione (2-methyl-1,4-naphthoquine), also known as vitamin K3, has been widely used as a model compound in the field of oxidative stress-related research. The metabolism of menadione has been studied, and it is known that menadione undergoes a two-electron reduction by NAD(P)H:Quinone oxidoreductase 1 (NQO1) after which the reduced form of menadione (2-methyl-1,4-naphthalenediol, menadiol) is glucuronidated and excreted in urine. To investigate which human UDP glucuronosyltransferase (UGT) isoforms participate in the glucuronidation of menadiol reduced by NQO1 from menadione, we first constructed heterologously expressed NQO1 in Sf9 cells and tested the menadiol glucuronidating activity of 16 human recombinant UGT isoforms. Of the 16 UGT isoforms, UGTs 1A6, 1A7, 1A8, 1A9, and 1A10 catalyzed menadiol glucuronidation, and, of these, UGTs 1A6 and 1A10 catalyzed menadiol glucuronidation at much higher rates than the other UGTs. Menadiol was regioselectively glucuronidated in the manner of 4-position>1 position by UGTs 1A7, 1A8, 1A9, and 1A10. In contrast to these UGTs, only UGT1A6 exhibited 1-menadiol-preferential glucuronidating activity. The results suggest possible detoxification pathways for quinones via NQO1 reduction followed by UGT glucuronidation. PMID- 18423375 TI - Involvement of 67-kDa laminin receptor-mediated myosin phosphatase activation in antiproliferative effect of epigallocatechin-3-O-gallate at a physiological concentration on Caco-2 colon cancer cells. AB - Previously we reported that 67-kDa laminin receptor (67LR) mediates epigallocatechin-3-O-gallate (EGCG)-induced cell growth inhibition and reduction of myosin regulatory light chain (MRLC) phosphorylation at Thr-18/Ser-19, which is important for cytokinesis. Here, we found that human colon adenocarcinoma Caco 2 cells exhibited higher expression level of 67LR and EGCG at a physiologically achievable concentration (1 microM) significantly accumulated the cells in G(2)/M phase without affecting expression of Wnt-signaling components. We also found that myosin phosphatase targeting subunit 1 (MYPT1) phosphorylation at Thr-696, which inhibits myosin phosphatase and promotes MRLC phosphorylation, was reduced in response to 1 microM EGCG. 67LR knockdown by RNA interference abolished the inhibitory effects of 1 microM EGCG on cell cycle progression and the phosphorylation of MRLC and MYPT1. These results suggest that through 67LR, EGCG at a physiological concentration can activate myosin phosphatase by reducing MYPT1 phosphorylation and that may be involved in EGCG-induced cell growth inhibition. PMID- 18423376 TI - Dendritic cell vaccine with mRNA targeted to the proteasome by polyubiquitination. AB - Dendritic cells (DCs) transfected with mRNA encoding tumor-associated antigens (TAAs) can induce tumor-specific T-cell responses. To potentiate this, we transfected mature DCs (mDCs) with mRNA encoding TAA targeted to the proteasome. DCs were generated from bone marrow cells by culture with 20 ng/ml GM-CSF and maturation with 1 microg/ml LPS. These mDCs were then electroporated with 10 microg of mRNA. Antigen presentation after electroporation with in vitro transcribed mRNA was compared with mRNA from a construct of the TAA preceded by ubiquitin. Proteasomal targeting of mRNA encoding cotranslationally ubiquitinated antigen was found to enhance intracellular degradation of target protein, and result in more efficient priming and expansion of TAA-specific CD8(+) T-cells. We therefore suggest that RNA-transfected DC vaccine efficacy could be improved by the use of mRNA targeted to the proteasome. PMID- 18423377 TI - Thiazolidinediones inhibit the growth of PC12 cells both in vitro and in vivo. AB - Thiazolidinediones (TZDs) have recently been proposed as a therapy for PPARgamma expressing tumors. Pheochromocytoma (PHEO) is associated with high morbidity and mortality due to excess catecholamine production, and few effective drug therapies currently exist. We investigated the effects of TZDs on PHEO both in vitro and in vivo. PPARgamma protein was expressed in human adrenal PHEO tissues as well as in rat PHEO cells, PC12. TZDs, including rosiglitazone (RGZ) and pioglitazone (PGZ), inhibited proliferation of PC12 cells in a dose-dependent manner and increased casapse-3 expression of PC12 cells. TZDs also reduced expression of cyclin E and cyclin-dependent kinase2. RGZ inhibited nerve growth factor-induced neurite outgrowth and reduced expression of catecholamine synthesizing enzymes. Finally, rat PHEO growth generated by subcutaneous injection of PC12 cells was slowed in an RGZ-treated mouse. These data suggest that TZDs may be a promising therapeutic approach for medical treatment for PHEO. PMID- 18423378 TI - Characterization of a stem cell population in lung cancer A549 cells. AB - We isolated a stem cell subpopulation from human lung cancer A549 cells using FACS/Hoechst 33342. This side population (SP), which comprised 24% of the total cell population, totally disappeared after treatment with the selective ABCG 2 inhibitor fumitremorgin C. In a repopulation study, isolated SP and non-SP cells were each able to generate a heterogeneous population of SP and non-SP cells, but this repopulation occurred more rapidly in SP cells than non-SP. An MTT assay and cell cycle distribution analysis reveal a similar profile between SP and non-SP groups. However, in the presence of doxorubicin (DOX) and methotrexate (MTX), SP cells showed significantly lower Annexin V staining when compared to non-SP cells. Taken together, these results demonstrate that SP cells have an active regeneration capacity and high anti-apoptotic activity compared with non-SP cells. Furthermore, our GeneChip data revealed a heightened mRNA expression of ABCG2 and ABCC2 in SP cells. Overall these data explain why the SP of A549 has a unique ability to resist DOX and MTX treatments. Therefore, we suggest that the expression of the ABCG2 transporter plays an important role in the multidrug resistance phenotype of A549 SP cells. PMID- 18423379 TI - Sphingosine 1-phosphate induces Mcl-1 upregulation and protects multiple myeloma cells against apoptosis. AB - Sphingosine 1-phosphate (S1P) is a bioactive lysophospholipid which is known to induce diverse cellular responses through at least five G-protein-coupled receptors on various cell types. However, neither the distribution of S1P receptors nor the effects of S1P on multiple myeloma (MM) cells are fully understood. Here, we show that MM cells express the S1P receptors, S1P1, S1P2, and S1P3. Furthermore, S1P protects MM cells against Dex-induced apoptosis. Importantly, S1P upregulates Mcl-1 expression in a time- and concentration dependent manner in human MM cell lines. Treatment of MM cells with pertussis toxin (PTX), a pan-S1P receptor inhibitor, results in blockage of S1P-induced upregulation of Mcl-1. These data demonstrate that S1P upregulates the expression of Mcl-1 and protects MM cells from Dex-induced apoptosis, providing the preclinical framework for novel therapeutics targeting at both Mcl-1 and/or S1P to improve the patient outcome in MM. PMID- 18423380 TI - Difference between Abbott AxSYM BNP and Triage BNP (Beckman Coulter DxI 800) concentrations. PMID- 18423381 TI - Modification of the oxidative stress biomarker AOPP assay: application in uremic samples. AB - BACKGROUND: The levels of advanced oxidation protein products (AOPP), a plasma protein biomarker used to assess oxidative stress, are elevated in patients with chronic kidney disease (CKD). However, this apparent elevation is to a large extent due to assay interference (mostly by triglycerides which are usually markedly elevated in CKD). We therefore developed and tested a modified version of the AOPP assay to minimize the impact of this interference. METHODS: Plasma levels of AOPP, lipids, proteins and various biomarkers of inflammation and oxidative stress were analyzed in 218 prevalent hemodialysis patients and 13 healthy controls using the established original (oAOPP) assay and following precipitation of plasma lipids using dextran sulphate (modified assay, mAOPP). The modified results were validated against a lipid extraction procedure using ether/butanol. RESULTS: The modified assay decreased the levels of triglycerides and AOPP by 87% and 38%, respectively. Whereas oAOPP values correlated strongly with triglycerides, no such correlation was seen with mAOPP. The mAOPP levels correlated significantly with the oxidative stress markers 8-oxo-dG and pentosidine, whereas no such correlations were found for oAOPP. CONCLUSIONS: The oAOPP concentration is largely overestimated in plasma samples due to lipid interferences. Precipitation of triglycerides before analysis yields markedly lower mAOPP values which more accurately reflect oxidative stress. Based on these results we propose that AOPP should be analyzed using the modified assay, which is a cheap, simple and fast method. PMID- 18423382 TI - The multiple functions of the thiol-based electron flow pathways of Escherichia coli: Eternal concepts revisited. AB - Electron flow via thiols is a theme with many variations in all kingdoms of life. The favourable physichochemical properties of the redox active couple of two cysteines placed in the optimised environment of the thioredoxin fold allow for two electron transfers in between top biological reductants and ultimate oxidants. The reduction of ribonucleotide reductases by thioredoxin and thioredoxin reductase of Escherichia coli (E. coli) was one of the first pathways to be elucidated. Diverse functions such as protein folding in the periplasm, maturation of respiratory enzymes, detoxification of hydrogen peroxide and prevention of oxidative damage may be based on two electron transfers via thiols. A growing field is the relation of thiol reducing pathways and the interaction of E. coli with different organisms. This concept combined with the sequencing of the genomes of different bacteria may allow for the identification of fine differences in the systems employing thiols for electron flow between pathogens and their corresponding mammalian hosts. The emerging possibility is the development of novel antibiotics. PMID- 18423383 TI - Brain oxidative stress in a triple-transgenic mouse model of Alzheimer disease. AB - Alzheimer disease (AD) is a neurodegenerative disease which is characterized by the presence of extracellular senile plaques mainly composed of amyloid-beta peptide (Abeta), intracellular neurofibrillary tangles, and selective synaptic and neuronal loss. AD brains revealed elevated levels of oxidative stress markers which have been implicated in Abeta-induced toxicity. In the present work we addressed the hypothesis that oxidative stress occurs early in the development of AD and evaluated the extension of the oxidative stress and the levels of antioxidants in an in vivo model of AD, the triple-transgenic mouse, which develops plaques, tangles, and cognitive impairments and thus mimics AD progression in humans. We have shown that in this model, levels of antioxidants, namely, reduced glutathione and vitamin E, are decreased and the extent of lipid peroxidation is increased. We have also observed increased activity of the antioxidant enzymes glutathione peroxidase and superoxide dismutase. These alterations are evident during the Abeta oligomerization period, before the appearance of Abeta plaques and neurofibrillary tangles, supporting the view that oxidative stress occurs early in the development of the disease. PMID- 18423384 TI - The impact of natural products upon modern drug discovery. AB - In the period 1970-2006, a total of 24 unique natural products were discovered that led to an approved drug. We analyze these successful leads in terms of drug like properties, and show that they can be divided into two equal subsets. The first falls in the 'Lipinski universe' and complies with the Rule of Five. The second is a 'parallel universe' that violates the rules. Nevertheless, the latter compounds remain largely compliant in terms of logP and H-bond donors, highlighting the importance of these two metrics in predicting bioavailability. Natural products are often cited as an exception to Lipinski's rules. We believe this is because nature has learned to maintain low hydrophobicity and intermolecular H-bond donating potential when it needs to make biologically active compounds with high molecular weight and large numbers of rotatable bonds. In addition, natural products are more likely than purely synthetic compounds to resemble biosynthetic intermediates or endogenous metabolites, and hence take advantage of active transport mechanisms. Interestingly, the natural product leads in the Lipinski and parallel universe had an identical success rate (50%) in delivering an oral drug. PMID- 18423385 TI - Characterization of the murine CTP:phosphocholine cytidylyltransferase beta gene promoter. AB - CTP:phosphocholine cytidylyltransferase (CCT) is a key regulatory enzyme in phosphatidylcholine (PtdCho) biosynthesis by the Kennedy pathway. In mammals, there are two genes that encode the enzyme isoforms that catalyze this reaction: Pcyt1a for CCTalpha and Pcyt1b for CCTbeta. In mouse tissues two different CCTbeta variants named CCTbeta2 and CCTbeta3 have been identified. Although little is known about Pcyt1b gene expression, recent data from cell lines propose a distinct role for CCTbeta2 in neuronal differentiation. Also, gonadal dysfunction in the CCTbeta2 knockout mouse suggests a role for this protein in ovary maturation and the maintenance of sperm production. This work defines and characterizes two alternative promoters that drive the expression of the two murine CCTbeta isoforms. The promoter activities were measured in Neuro-2a (mouse neuroblastoma), TM4 (mouse Sertoli) and C3H10T1/2 (mouse embryo fibroblast) cell lines. The transcriptional start points of each transcript and the promoter regions essential for the expression of each isoform were determined. Analysis of the CCTbeta2 promoter sequence suggested the transcription factor AP-1 as a potential regulator of CCTbeta2 expression in neuronal cells. However, CCTbeta3 was not detected in this cell line suggesting a different role or regulation. The activities of alternative promoters provide for greater flexibility in the control of CCTbeta isoform expression. PMID- 18423386 TI - Potential role of phospholipase D2 in increasing interleukin-2 production by T lymphocytes through activation of mitogen-activated protein kinases ERK1/ERK2. AB - Hydrolysis of phosphatidylcholine by phospholipase D (PLD) leads to the generation of phosphatidic acid (PA), which is itself a source of diacylglycerol (DAG). These two versatile lipid second messengers are at the centre of a phospholipid signalling network and as such are involved in several cellular functions. However, their role in T-cell activation and functions are still enigmatic. In order to elucidate this role, we generated a human and a murine T cell line that stably overexpressed the PLD2 isoform. Analysis of the Ras-MAPK pathway upon phorbol myristate acetate (PMA) and ionomycin stimulation revealed that PLD2 promoted an early and sustained increase in ERK1/2 phosphorylation in both cell lines. This response was inhibited by 1-butanol, a well known distracter of PLD activity, or upon overexpression of a dominant negative PLD2, and it was concomitant with a boost of PA/DAG production. As a functional consequence of this PLD2-dependent MAPK activation, interleukin-2 production evoked by PMA/ionomycin stimulation or CD3/CD28 engagement was enhanced in the two T-cell lines overexpressing PLD2. Thus, PLD2 emerged as an early player upstream of the Ras-MAPK-IL-2 pathway in T-cells via PA and DAG production, raising new possibilities of pharmacological manipulation in immune disorders. PMID- 18423388 TI - Electrochemical immunosensor with aptamer-based enzymatic amplification. AB - An electrochemical immunosensor is reported by using aptamer-based enzymatic amplification with immunoglobulin E (IgE) as the model analyte. In this method, the IgE antibody is covalently immobilized as the capture probe on the gold electrode via a self-assembled monolayer of cysteamine. After the target is captured, the biotinylated anti-IgE aptamer is used as the detection probe. The specific interaction of streptavidin-conjugated alkaline phosphatase to the surface-bound biotinylated detection probe mediates a catalytic reaction of ascorbic acid 2-phosphate substrate to produce a reducing agent ascorbic acid. Then silver ions in the solution can be reduced, leading to the deposition of metallic silver on the electrode surface. The amount of deposited silver, which is determined by the amount of IgE target bound on the electrode surface, can be quantified using the stripping voltammetry. The results obtained demonstrated that the electrochemical immunosensor possesses high specificity and a wide dynamic range with a low detection limit that possibly arises from the combination of the highly specific aptamer and the highly sensitive stripping determination of enzymatically deposited silver. PMID- 18423389 TI - A dehydrochlorinase-based pH change assay for determination of DDT in sprayed surfaces. AB - A glutathione S-transferase (GST) from the mosquito Aedes aegypti (aagste2), selected in the field as a major metabolic resistance enzyme for this parasite vector, was employed to produce a highly specific assay for the determination of DDT [1,1,1-dichloro-2,2-bis(p-chlorophenyl)ethylene]. Detection is based on the pH change occurring in an appropriate buffer system by the concomitant release of H(+) during the aagste2-catalyzed dehydrochlorination reaction and is monitored potentiometrically or colorimetrically in the presence of a pH marker. The theoretical limit of detection (LOD) of the assay is 3.8 microg/ml, and the linear range of quantification is 12 to 250 microg/ml. The method does not recognize biologically inactive DDT analogues or major DDT photodegradants and breakdown molecules, and it is highly specific for the insecticidal p.p'DDT [1,1,1-trichloro-2,2-bis(p-chlorophenyl) ethane]. The biosensor was validated with a number of insecticide swabs from DDT-sprayed surfaces and found to be reproducible and reliable as compared with high-performance liquid chromatography (HPLC) (correlation coefficient R(2)=0.98). Given the current expansion of DDT residual sprayings in many regions of Africa as a key strategic intervention for malaria vector control, this simple assay to monitor DDT levels for vector control spraying programs could have an important impact on malaria control. PMID- 18423387 TI - TNF-alpha stimulation inhibits siRNA-mediated RNA interference through a mechanism involving poly-(A) tail stabilization. AB - The control of mRNA stability is a complex biological process that involves numerous factors, including microRNA (miRNA) and short interfering RNA (siRNA). Here, we show that short interfering RNA (siRNA) and microRNA share some similarities in their response to cellular stress. miR16 expedites the degradation of mRNAs containing AU-rich elements (ARE) in their 3' untranslated region (UTR). si20 is an siRNA designed to target a non-ARE sequence in the TNF 3'UTR. We found that both si20 and miR16/ARE-mediated degradation of mRNAs can be inhibited by stimulating cells with different stresses. By analyzing TNF-alpha stimulation-mediated stabilization of si20- and miR16-targeted mRNA, we show that this stabilization is not caused by modifying si20 and miR16 loading into Ago2 complexes, or mRNA targeting to Ago2, but by inhibiting mRNA deadenylation. This is the first report showing that a specific siRNA-mediated mRNA degradation can be regulated by inflammatory stimuli, and that deadenylation is involved in this siRNA-mediated mRNA decay. PMID- 18423390 TI - C18 ceramide analysis in mammalian cells employing reversed-phase high performance liquid chromatography tandem mass spectrometry. AB - Ceramides play an important role in diverse cellular functions such as differentiation, cell cycle progression, cell-cell adhesion, senescence, and apoptosis. Here we report a method of extracting lipids from mammalian cells and quantifying ceramide, where the assay conditions were optimized for reproducibility, linearity, recovery, and sensitivity. Simultaneous chromatographic separations were carried out by reversed-phase high-performance liquid chromatography coupled to electrospray ionization using a Pursuit 3 Diphenyl column (50 x 2.0 mm) and supported by a mobile phase consisting of acetonitrile plus 0.1% formic acid and 25 mM ammonium acetate. Ceramides were detected in the multiple reaction mode by tandem mass spectrometry in the positive ion mode, and all extracted ion peaks were integrated for quantitative analysis. The limits of detection and quantification achieved were 0.2 and 1.0 pg on column, respectively. Using this method, we successfully quantified and compared differences in C(18) ceramide levels induced by two DNA-damaging agents, mitomycin C and daunorubicin, and two apoptosis-inducing ligands, tumor necrosis factor alpha (TNF-alpha) and TNF-related apoptosis-inducing ligand (TRAIL). This work, therefore, describes a method that will be helpful for investigating how ceramide is regulated by different chemotherapeutic agents and will help us to better understand the mechanisms of signal transduction involving ceramide. PMID- 18423391 TI - Distinct organization of energy metabolism in HL-1 cardiac cell line and cardiomyocytes. AB - Expression and function of creatine kinase (CK), adenylate kinase (AK) and hexokinase (HK) isoforms in relation to their roles in regulation of oxidative phosphorylation (OXPHOS) and intracellular energy transfer were assessed in beating (B) and non-beating (NB) cardiac HL-l cell lines and adult rat cardiomyocytes or myocardium. In both types of HL-1 cells, the AK2, CKB, HK1 and HK2 genes were expressed at higher levels than the CKM, CKMT2 and AK1 genes. Contrary to the saponin-permeabilized cardiomyocytes the OXPHOS was coupled to mitochondrial AK and HK but not to mitochondrial CK, and neither direct transfer of adenine nucleotides between CaMgATPases and mitochondria nor functional coupling between CK-MM and CaMgATPases was observed in permeabilized HL-1 cells. The HL-1 cells also exhibited deficient complex I of the respiratory chain. In conclusion, contrary to cardiomyocytes where mitochondria and CaMgATPases are organized into tight complexes which ensure effective energy transfer and feedback signaling between these structures via specialized pathways mediated by CK and AK isoforms and direct adenine nucleotide channeling, these complexes do not exist in HL-1 cells due to less organized energy metabolism. PMID- 18423392 TI - Structure and regulation of the vacuolar ATPases. AB - The vacuolar (H(+))-ATPases (V-ATPases) are ATP-dependent proton pumps responsible for both acidification of intracellular compartments and, for certain cell types, proton transport across the plasma membrane. Intracellular V-ATPases function in both endocytic and intracellular membrane traffic, processing and degradation of macromolecules in secretory and digestive compartments, coupled transport of small molecules such as neurotransmitters and ATP and in the entry of pathogenic agents, including envelope viruses and bacterial toxins. V-ATPases are present in the plasma membrane of renal cells, osteoclasts, macrophages, epididymal cells and certain tumor cells where they are important for urinary acidification, bone resorption, pH homeostasis, sperm maturation and tumor cell invasion, respectively. The V-ATPases are composed of a peripheral domain (V(1)) that carries out ATP hydrolysis and an integral domain (V(0)) responsible for proton transport. V(1) contains eight subunits (A-H) while V(0) contains six subunits (a, c, c', c'', d and e). V-ATPases operate by a rotary mechanism in which ATP hydrolysis within V(1) drives rotation of a central rotary domain, that includes a ring of proteolipid subunits (c, c' and c''), relative to the remainder of the complex. Rotation of the proteolipid ring relative to subunit a within V(0) drives active transport of protons across the membrane. Two important mechanisms of regulating V-ATPase activity in vivo are reversible dissociation of the V(1) and V(0) domains and changes in coupling efficiency of proton transport and ATP hydrolysis. This review focuses on recent advances in our lab in understanding the structure and regulation of the V-ATPases. PMID- 18423393 TI - Prevention of leak in the proton pump of cytochrome c oxidase. AB - The cytochrome c oxidases (CcO), which are responsible for most O(2) consumption in biology, are also redox-linked proton pumps that effectively convert the free energy of O(2) reduction to an electrochemical proton gradient across mitochondrial and bacterial membranes. Recently, time-resolved measurements have elucidated the sequence of events in proton translocation, and shed light on the underlying molecular mechanisms. One crucial property of the proton pump mechanism has received less attention, viz. how proton leaks are avoided. Here, we will analyse this topic and demonstrate how the key proton-carrying residue Glu-242 (numbering according to the sequence of subunit I of bovine heart CcO) functions as a valve that has the effect of minimising back-leakage of the pumped proton. PMID- 18423394 TI - Sorting and assembly of mitochondrial outer membrane proteins. AB - In the last years the picture of protein import into the mitochondria has become much more complicated in terms of new components and new sorting pathways. These novel findings have also changed views concerning the biogenesis pathway of mitochondrial outer membrane proteins. In addition to proteins anchored with transmembrane alpha-helices, the endosymbiotic origin of the mitochondria has resulted in the presence of transmembrane beta-barrels in this compartment. The sorting and assembly pathway of outer membrane proteins involves three machineries: the translocase of the outer membrane (TOM complex) the sorting and assembly machinery (SAM complex) and the MDM complex (mitochondrial distribution and morphology). Here we review recent developments on the biogenesis pathways of outer membrane proteins with a focus on Tom proteins, the most intensively studied class of these precursor proteins. PMID- 18423396 TI - Isoform-specific regulation of adipocyte differentiation by Akt/protein kinase Balpha. AB - The phosphatidylinositol 3-kinase (PI3K)/Akt pathway tightly regulates adipose cell differentiation. Here we show that loss of Akt1/PKBalpha in primary mouse embryo fibroblast (MEF) cells results in a defect of adipocyte differentiation. Adipocyte differentiation in vitro and ex vivo was restored in cells lacking both Akt1/PKBalpha and Akt2/PKBbeta by ectopic expression of Akt1/PKBalpha but not Akt2/PKBbeta. Akt1/PKBalpha was found to be the major regulator of phosphorylation and nuclear export of FoxO1, whose presence in the nucleus strongly attenuates adipocyte differentiation. Differentiation-induced cell division was significantly abrogated in Akt1/PKBalpha-deficient cells, but was restored after forced expression of Akt1/PKBalpha. Moreover, expression of p27(Kip1), an inhibitor of the cell cycle, was down regulated in an Akt1/PKBalpha specific manner during adipocyte differentiation. Based on these data, we suggest that the Akt1/PKBalpha isoform plays a major role in adipocyte differentiation by regulating FoxO1 and p27(Kip1). PMID- 18423395 TI - Forty years of Mitchell's proton circuit: From little grey books to little grey cells. AB - It is more than forty years since Peter Mitchell published his first 'little grey book' laying out his chemiosmotic hypothesis. Although ideas about the molecular mechanisms of the proton pumps have evolved considerably since then, his concept of 'coupling through proton circuits' remains remarkably prescient, and has provided the inspiration for the research careers of this author and many others. This review is a personal account of how the proton circuit has been followed from the little grey book, via brown fat and calcium transport to investigations into the life and death of neurons, Hercule Poirot's 'little grey cells'. PMID- 18423397 TI - Shared traits on the reaction coordinates of ribonuclease and an RNA enzyme. AB - Reaction-intermediate analogs have been used to understand how phosphoryl transfer enzymes promote catalysis. Herein we report the first structure of a small ribozyme crystallized with a 3'-OH, 2',5'-linkage in lieu of the normal phosphodiester substrate. The new structure shares features of the reaction coordinate exhibited in prior ribozyme structures including a vanadate complex that mimicked the oxyphosphorane transition state. As such, the structure exhibits reaction-intermediate traits that allow direct comparison of stabilizing interactions to the 3'-OH, 2',5'-linkage contributed by the RNA enzyme and its protein counterpart, ribonuclease. Clear similarities are observed between the respective structures including hydrogen bonds to the non-bridging oxygens of the scissile phosphate. Other commonalities include carefully poised water molecules that may alleviate charge build-up in the transition state and placement of a positive charge near the leaving group. The advantages of 2',5'-linkages to investigate phosphoryl-transfer reactions are discussed, and argue for their expanded use in structural studies. PMID- 18423398 TI - The changing scenario in laboratory medicine and the role of laboratory professionals in translational medicine. AB - A revolution is now expected to occur in the ever-changing scenario of laboratory medicine thanks to the introduction of "omics" into clinical practice. However, our awareness of the divide between knowledge and practice, and the understandable assumption that omics research will be riddled with difficulties, has led to the concepts of knowledge translation, translational research and translational medicine. The interchangeable terms, translational research or translational medicine (hereafter referred to as translational research), currently underline the pressing need to gain practical benefit from the enormous investments made in biomedical research by the private and public sector. From the viewpoint of physicians, clinical laboratory professionals and patients, who are more directly involved in clinical practice, translational research responds to the need to obtain benefit from research, thus closing the gap between what we know and what we practice. This means transferring diagnostic and therapeutic advances that have proven effective in large well-conducted trials (and are therefore evidenced-based,) to daily medical practice. Translational research should be regarded as a two-way road: bench to bedside, and bedside to bench. However, to achieve a more effective translation process, a new road map should be implemented through interaction and cooperation between basic researchers, clinicians, laboratory professionals and manufacturers. Some examples of recent developments in clinical laboratory testing, including markers of cardiovascular diseases, clinical proteomics and recombinant allergens, attest to the importance of a careful evaluation of all variables allowing the introduction of such new insights into clinical practice in order to assure better clinical outcomes. The ability of laboratory medicine to deliver safer and more effective health care calls for a more careful evaluation not only of analytical characteristics but also of any other variables that may affect the clinical usefulness and diagnostic performances of laboratory tests thus enabling a more accurate interpretation and utilization of laboratory information. PMID- 18423399 TI - Development of an optimized multimarker strategy for early risk assessment of patients with acute coronary syndromes. AB - BACKGROUND: A multitude of biomarkers have been suggested for early risk assessment in patients admitted to the emergency department with suspected acute coronary syndromes. We used logistic regression synergistically with classification and regression tree (CART) analysis to define a multimarker strategy and the cut-off values and sequencing needed to optimize risk stratification in a low to moderate risk population of the emergency department. METHODS: 432 unselected patients (59.7+/-14.5 y, 60.4% male) admitted to the emergency department (ED) with acute coronary syndromes (ACS) were enrolled. Cardiac troponin I (cTnI), N-terminal pro-B-Type natriuretic peptide (NT-proBNP), high sensitivity C-reactive protein (hsCRP), placental growth factor (PlGF), lipoprotein-associated phospholipase A(2) (Lp-PLA(2)) and D-dimers were measured by immunoassay and whole blood choline (WBCHO) and plasma choline (PLCHO) were measured using LC/MS from baseline samples. Logistic regression and CART analysis were used to define the importance of the various biomarkers tested and to define their hierarchy with respect to the prediction of major adverse cardiac events (MACE; cardiac death, non-fatal MI, unstable angina, CHF requiring admission, urgent PCI and CABG) over the 42-day follow-up period. RESULTS: A combination of NT-proBNP, WBCHO and Lp-PLA2 with cutoffs identified by CART-analysis was optimal for risk-stratification and superior to all other possible combinations of markers. Increased concentrations of both NT-proBNP (>1400 ng/l) and WBCHO (>21 micromol/l) identified patients with very high risk (RR=2.4, 39% primary endpoint) while low concentrations of NT-proBNP (< or = 1400 ng/l), WBCHO (< or = 17 micromol/l) and LP-PLA2 (< or = 210 microg/l) indicated very low risk (0% primary endpoint). WBCHO > 17 micromol/l additionally identified a subgroup with intermediate risk (RR=3.0, 13.5% primary endpoint) in patients with NT-proBNP concentrations < or = 1400 ng/l. Troponin when increased was highly prognostic but was not often positive in this early cohort. CONCLUSIONS: A multimarker strategy defined synergistically by logistic regression and by classification and regression tree (CART) analysis can stratify patients into risk groups ranging from very low risk (0% MACE) to very high risk (39.5% MACE) based on admission values. PMID- 18423400 TI - The amino- and carboxyterminal cross-linked telopeptides of collagen type I, NTX I and CTX-I: a comparative review. AB - Bone diseases such as osteoporosis or bone metastases are a continuously growing problem in the ageing populations across the world. In recent years, great efforts have been made to develop specific and sensitive biochemical markers of bone turnover that could help in the assessment and monitoring of bone turnover. The amino- and carboxyterminal cross-linked telopeptides of type I collagen (NTX I and CTX-I, respectively) are two widely used bone resorption markers that attracted great attention due to their relatively high sensitivity and specificity for the degradation of type I collagen, and their rapid adaptation to automated analyzers. However, the clinical performance of both markers differs significantly depending on the clinical situation. These differences have caused considerable confusion and uncertainty. If used correctly, both markers have great potential to improve the management of many bone diseases. We here review the biochemistry, analytical background and clinical performance of NTX-I and CTX I, as documented in the accessible literature until March 2008. PMID- 18423401 TI - Training and education of laboratory medicine professionals. Foreword. PMID- 18423402 TI - Paraoxonase 1 (PON1) status in gastroesophageal malignancies and associated paraneoplastic syndromes - connection with inflammation. AB - OBJECTIVE: To evaluate the status and diagnostic utility of antioxidant paraoxonase 1 (PON1) in gastroesophageal cancers. DESIGN AND METHODS: PON1's arylesterase/paraoxonase activities and phenotype were determined in 82 cancers and 57 controls, and related to clinicopathological features, anemia and cachexia coexistence, cholesterol levels, liver function panel, and inflammatory and angiogenic indices. RESULTS: PON1's activities were decreased in cancer. Arylesterase activity correlated with cancer T and N stages, being an N1 independent predictor. Both activities correlated with transferrin, albumin, CRP and inflammation-based Glasgow Prognostic Score, and arylesterase activity also with interleukin-6 and midkine. Phenotype A (Q192 homozygotes) was associated with lower transferrin and higher TNF-alpha concentrations. PON1's arylesterase activity reflected anemia severity, being correlated with hemoglobin, hematocrit, and iron. PON1 arylesterase activity negatively correlated with alkaline phosphatase and gamma-glutamyl transferase, but not with bilirubin, aminotransferases, HDL or LDL cholesterol. PON1 moderately indicated cancer presence and regional metastasis. CONCLUSIONS: PON1 activity decreases in gastroesophageal cancers and corresponds to inflammation severity and cancer related anemia. PON1 decrease indicates lymph node metastasis, but its moderate predictive power does not recommend PON1 determination alone for clinical application. PMID- 18423403 TI - Adiponectin promotes endothelial progenitor cell number and function. AB - Obesity-linked diseases are associated with suppressed endothelial progenitor cell (EPC) function. Adiponectin is an adipose-derived protein that is downregulated in obese and diabetic subjects. Here, we investigated the effects of adiponectin on EPCs. EPC levels did not increase in adiponectin deficient (APN KO) in response to hindlimb ischemia. Adenovirus-mediated delivery of adiponectin increased EPC levels in both WT and APN-KO mice. Incubation of human peripheral blood mononuclear cells with adiponectin led to an increase of the number of EPCs. Adiponectin induced EPC differentiation into network structures and served as a chemoattractant in EPC migration assays. These data suggest that hypoadiponectinemia may contribute to the depression of EPC levels that are observed in patients with obesity-related cardiovascular disorders. PMID- 18423404 TI - Nuclear actin dynamics--from form to function. AB - Cell biological functions of actin have recently expanded from cytoplasm to nucleus, with actin implicated in such diverse processes as gene expression, transcription factor regulation and intranuclear motility. Actin in the nucleus seems to behave differently than in the cytoplasm, raising new questions regarding the molecular mechanisms by which actin functions in cells. In this review, I will discuss dynamic properties of nuclear actin that are related to its polymerization cycle and nucleocytoplasmic shuttling. By comparing the behaviour of nuclear and cytoplasmic actin and their regulators, I try to dissect the underlying differences of these equally important cellular actin pools. PMID- 18423405 TI - ROCK and PRK-2 mediate the inhibitory effect of Y-27632 on polyglutamine aggregation. AB - Polyglutamine expansion in huntingtin (Htt) and the androgen receptor (AR) causes untreatable neurodegenerative diseases. Y-27632, a therapeutic lead, reduces Htt and AR aggregation in cultured cells, and Htt-induced neurodegeneration in Drosophila. Y-27632 inhibits both Rho-associated kinases ROCK and PRK-2, making its precise intracellular target uncertain. Over-expression of either kinase increases Htt and AR aggregation. Three ROCK inhibitors (Y-27632, HA-1077, and H 1152P), and a specific ROCK inhibitory peptide reduce polyglutamine protein aggregation, as does knockdown of ROCK or PRK-2 by RNAi. RNAi also indicates that each kinase is required for the inhibitory effects of Y-27632 to manifest fully. These two actin regulatory kinases are thus involved in polyglutamine aggregation, and their simultaneous inhibition may be an important therapeutic goal. PMID- 18423406 TI - Aggregation of proteins having Golgi apparatus sorting determinant induces large globular structures derived from the endoplasmic reticulum in plant seed cells. AB - Endoplasmic reticulum (ER)-derived compartments are found in many plant species. Although it has been assumed that aggregation induces formation of the ER-derived compartments in plant seed cells, the effect of aggregation on the trafficking from the ER to the Golgi has not yet been elucidated. In this study, we used an aggregated type of red fluorescent protein (DsRED) to investigate the effect of aggregation on sorting in seed cells. DsRED fused to the Golgi sorting determinant was found mainly in large globular structures derived from the ER where ER-resident proteins were excluded. These results indicate that aggregation of the Golgi protein blocks transport from the ER to the Golgi. PMID- 18423409 TI - 2,4-Diamino-6-hydroxypyrimidine (DAHP) suppresses cytokine-induced VCAM-1 expression on the cell surface of human umbilical vein endothelial cells in a BH(4)-independent manner. AB - 2,4-Diamino-6-hydroxypyrimidine (DAHP) is considered a specific inhibitor of BH(4) biosynthesis and is widely used in order to elucidate the possible biological function of BH(4) in various cells. In the present study, we found that both the synthesis of tetrahydrobiopterin (BH(4)) and expression of vascular cell adhesion molecule 1 (VCAM-1) were increased in human umbilical vein endothelial cells (HUVEC) treated with proinflammatory cytokines. Thus we examined the effects of DAHP to clarify whether BH(4) might be involved in the expression of VCAM-1 in HUVEC. DAHP reduced the levels of both BH(4) and VCAM-1 induced by TNF-alpha and IFN-gamma. However, the dose-response curves of DAHP for the suppression of the VCAM-1 level and that of BH(4) level were markedly different. Supplementation with sepiapterin failed to restore the depressed VCAM 1 level, although it completely restored the BH(4) level. Furthermore, DAHP significantly reduced the VCAM-1 level under the experimental conditions using TNF-alpha alone, which failed to induce BH(4) production. Taken together, these results indicate that DAHP inhibited the expression of VCAM-1 in a BH(4) independent manner in HUVEC. In the present study, we also found that DAHP significantly suppressed the accumulation of cytokine-induced NF-kappaB (p65) in the nucleus as well as the mRNA levels of VCAM-1 and GTP cyclohydrolase I (GTPCH), the rate-limiting enzyme of BH(4) synthesis. The data obtained in this study suggest that DAHP reduced VCAM-1 and GTPCH protein synthesis at least partially via suppressing the NF-kappaB level in the nucleus of HUVEC. PMID- 18423408 TI - Cholesterol homeostasis and the escape tendency (activity) of plasma membrane cholesterol. AB - We review evidence that sterols can form stoichiometric complexes with certain bilayer phospholipids, and sphingomyelin in particular. These complexes appear to be the basis for the formation of condensed and ordered liquid phases, (micro)domains and/or rafts in both artificial and biological membranes. The sterol content of a membrane can exceed the complexing capacity of its phospholipids. The excess, uncomplexed membrane sterol molecules have a relatively high escape tendency, also referred to as fugacity or chemical activity (and, here, simply activity). Cholesterol is also activated when certain membrane intercalating amphipaths displace it from the phospholipid complexes. Active cholesterol projects from the bilayer and is therefore highly susceptible to attack by cholesterol oxidase. Similarly, active cholesterol rapidly exits the plasma membrane to extracellular acceptors such as cyclodextrin and high-density lipoproteins. For the same reason, the pool of cholesterol in the ER (endoplasmic reticulum) increases sharply when cell surface cholesterol is incremented above the physiological set-point; i.e., equivalence with the complexing phospholipids. As a result, the escape tendency of the excess cholesterol not only returns the plasma membrane bilayer to its set-point but also serves as a feedback signal to intracellular homeostatic elements to down-regulate cholesterol accretion. PMID- 18423407 TI - Alteration of Akt activity increases chemotherapeutic drug and hormonal resistance in breast cancer yet confers an achilles heel by sensitization to targeted therapy. PMID- 18423410 TI - Hispolon induces apoptosis in human gastric cancer cells through a ROS-mediated mitochondrial pathway. AB - Severe side effects and complications such as gastrointestinal and hematological toxicities because of current anticancer drugs are major problems in the clinical management of gastric cancer, which highlights the urgent need for novel effective and less toxic therapeutic approaches. Hispolon, an active polyphenol compound, is known to possess potent antineoplastic and antiviral properties. In this study, we investigated the efficacy of hispolon in human gastric cancer cells and explored the cell death mechanism. Hispolon induced ROS-mediated apoptosis in gastric cancer cells and was more toxic toward gastric cancer cells than toward normal gastric cells, suggesting greater susceptibility of the malignant cells. The mechanism of hispolon-induced apoptosis was that hispolon abrogated the glutathione antioxidant system and caused massive ROS accumulation in gastric cancer cells. Excessive ROS caused oxidative damage to the mitochondrial membranes and impaired the membrane integrity, leading to cytochrome c release, caspase activation, and apoptosis. Furthermore, hispolon potentiated the cytotoxicity of chemotherapeutic agents used in the clinical management of gastric cancer. These results suggest that hispolon could be useful for the treatment of gastric cancer either as a single agent or in combination with other anticancer agents. PMID- 18423412 TI - Inhibition of NADPH oxidase-related oxidative stress-triggered signaling by honokiol suppresses high glucose-induced human endothelial cell apoptosis. AB - Angiopathy is a major complication of diabetes. Abnormally high blood glucose is a crucial risk factor for endothelial cell damage. Nuclear factor-kappaB (NF kappaB) has been demonstrated as a mediated signaling in hyperglycemia or oxidative stress-triggered apoptosis of endothelial cells. Here we explored the efficacy of honokiol, a small molecular weight natural product, on NADPH oxidase related oxidative stress-mediated NF-kappaB-regulated signaling and apoptosis in human umbilical vein endothelial cells (HUVECs) under hyperglycemic conditions. The methods of morphological Hoechst staining and annexin V/propidium iodide staining were used to detect apoptosis. Submicromolar concentrations of honokiol suppressed the increases of NADPH oxidase activity, Rac-1 phosphorylation, p22(phox) protein expression, and reactive oxygen species production in high glucose (HG)-stimulated HUVECs. The degradation of IkappaBalpha and increase of NF-kappaB activity were inhibited by honokiol in HG-treated HUVECs. Moreover, honokiol (0.125-1 microM) also suppressed HG-induced cyclooxygenase (COX)-2 upregulation and prostaglandin E(2) production in HUVECs. Honokiol could reduce increased caspase-3 activity and the subsequent apoptosis and cell death triggered by HG. These results imply that inhibition of NADPH oxidase-related oxidative stress by honokiol suppresses the HG-induced NF-kappaB-regulated COX-2 upregulation, apoptosis, and cell death in HUVECs, which has the potential to be developed as a therapeutic agent to prevent hyperglycemia-induced endothelial damage. PMID- 18423413 TI - Prevention of estrogen-DNA adduct formation in MCF-10F cells by resveratrol. AB - Resveratrol (Resv), a natural occurring phytolexin present in grapes and other foods, possesses chemopreventive effects revealed by its striking modulation of diverse cellular events associated with tumor initiation, promotion, and progression. Catechol estrogens generated in the metabolism of estrogens are oxidized to catechol quinones that react with DNA to form predominantly depurinating estrogen-DNA adducts. This event can generate the mutations responsible for cancer initiation. In this regard, Resv acts as both an antioxidant and an inducer of the phase II enzyme NAD(P)H:quinone oxidoreductase 1 (NQO1). In this report, we present the effects of Resv on the metabolism of estrogens in normal breast epithelial cells (MCF-10F) treated with 4 hydroxyestradiol (4-OHE(2)) or estradiol-3,4-quinone (E(2)-3,4-Q). Resv induced NQO1 in a dose- and time-dependent manner, but did not affect the expression of catechol-O-methyltransferase. Ultraperformance liquid chromatography/tandem mass spectrometry was used to determine the effects of Resv on estrogen metabolism. Preincubation of the cells with Resv for 48 h decreased the formation of depurinating estrogen-DNA adducts from 4-OHE(2) or E(2)-3,4-Q and increased formation of methoxycatechol estrogens. When Resv was also present with the 4 OHE(2) or E(2)-3,4-Q, even greater increases in methoxycatechol estrogens were observed, and the DNA adducts were undetectable. We conclude that Resv can protect breast cells from carcinogenic estrogen metabolites, suggesting that it could be used in breast cancer prevention. PMID- 18423411 TI - Redox-based regulation of signal transduction: principles, pitfalls, and promises. AB - Oxidants are produced as a by-product of aerobic metabolism, and organisms ranging from prokaryotes to mammals have evolved with an elaborate and redundant complement of antioxidant defenses to confer protection against oxidative insults. Compelling data now exist demonstrating that oxidants are used in physiological settings as signaling molecules with important regulatory functions controlling cell division, migration, contraction, and mediator production. These physiological functions are carried out in an exquisitely regulated and compartmentalized manner by mild oxidants, through subtle oxidative events that involve targeted amino acids in proteins. The precise understanding of the physiological relevance of redox signal transduction has been hampered by the lack of specificity of reagents and the need for chemical derivatization to visualize reversible oxidations. In addition, it is difficult to measure these subtle oxidation events in vivo. This article reviews some of the recent findings that illuminate the significance of redox signaling and exciting future perspectives. We also attempt to highlight some of the current pitfalls and the approaches needed to advance this important area of biochemical and biomedical research. PMID- 18423414 TI - Oxidative damage to extracellular matrix and its role in human pathologies. AB - The extracellular compartments of most biological tissues are significantly less well protected against oxidative damage than intracellular sites and there is considerable evidence for such compartments being subject to a greater oxidative stress and an altered redox balance. However, with some notable exceptions (e.g., plasma and lung lining fluid) oxidative damage within these compartments has been relatively neglected and is poorly understood. In particular information on the nature and consequences of damage to extracellular matrix is lacking despite the growing realization that changes in matrix structure can play a key role in the regulation of cellular adhesion, proliferation, migration, and cell signaling. Furthermore, the extracellular matrix is widely recognized as being a key site of cytokine and growth factor binding, and modification of matrix structure might be expected to alter such behavior. In this paper we review the potential sources of oxidative matrix damage, the changes that occur in matrix structure, and how this may affect cellular behavior. The role of such damage in the development and progression of inflammatory diseases is discussed. PMID- 18423415 TI - Dissimilarity-based approaches to compound acquisition. AB - The concept of molecular diversity has been integrated in drug discovery efforts for many years. Applications of molecular diversity have been used to identify compounds for screening and to select compounds to augment proprietary collections. These early efforts were crude and suffered from a number of faults, but their evolution has, over the years, led to an improvement in the computational procedures used to identify new commercial compounds for acquisition. Although not much has recently been written about modern methods for augmenting compound collections, this activity is still a very relevant and important task to those involved with the development of compound collections. This review focuses on the process and software used to identify compounds deemed worthy of acquisition. PMID- 18423416 TI - Coverage and bias in chemical library design. AB - The design of chemical libraries directed to target classes is an activity that requires the availability of ligand pharmacological data and/or protein structural data. On the basis of the knowledge derived from these data, chemical libraries directed mainly to G protein-coupled receptors, kinases, proteases, and nuclear receptors have been assembled. However, current design strategies widely overlook assessing the potential ability of the compounds contained in a focused library to provide uniform ample coverage of the protein family they intend to target. Here, we discuss the use of in silico target profiling methods as a means to estimate the actual scope of chemical libraries to probe entire protein families and illustrate its applicability in optimizing the composition of compound sets to achieve maximum coverage of the family with minimum bias to particular targets. PMID- 18423417 TI - Peptidomimetics, a synthetic tool of drug discovery. AB - The demand for modified peptides with improved stability profiles and pharmacokinetic properties is driving extensive research effort in this field. Many structural modifications of peptides guided by rational design and molecular modeling have been established to develop novel synthetic approaches. Recent advances in the synthesis of conformationally restricted building blocks and peptide bond isosteres are discussed. PMID- 18423418 TI - High glucose downregulates endothelial progenitor cell number via SIRT1. AB - Increasing evidence indicates that mammalian SIRT1 mediates calorie restriction and influences lifespan regulating a number of biological molecules such as FoxO1. SIRT1 controls the angiogenic activity of endothelial cells via deacetylation of FoxO1. Endothelial dysfunction and reduced new blood vessel growth in diabetes involve a decreased bioactivity of endothelial progenitor cells (EPCs) via repression of FoxO1 transcriptional activity. The relative contribution of SIRT1 with respect to the direct effects of high glucose on EPC number is poorly understood. We report that treatment of EPCs with high glucose for 3 days determined a consistent downregulation of EPC positive to DiLDL/lectin staining and, interestingly, this was associated with reduced SIRT1 expression levels and enzyme activity, and increased acetyl-FoxO1 expression levels. Moreover, EPCs responded to high glucose with major changes in the expression levels of cell metabolism-, cell cycle-, and oxidative stress-related genes or proteins. Proteomic analysis shows increased expression of nicotinamide phosphoribosyl transferase and mitochondrial superoxide dismutase whereas a glucose-related heat shock protein is reduced. These findings show that SIRT1 is a critical modulator of EPCs dysfunction during alteration of glucose metabolism. PMID- 18423419 TI - Cellular localization and expression of gp63 homologous metalloproteases in Leishmania (Viannia) braziliensis strains. AB - Leishmania (Viannia) braziliensis is the major causative agent of American tegumentary leishmaniasis, a disease that encompasses a broad spectrum of clinical manifestations. In a previous study, we showed that Brazilian and Colombian L. braziliensis strains, isolated from patients with distinct clinical manifestations, display different pattern of metalloprotease activities. Following these results, we investigated the cellular localization of these molecules and their relation to the major surface protease (gp63) of Leishmania. Comparative analyses of metalloprotease expression among different clinical isolates as well as an evaluation of the effect of long-term in vitro passage on the expression pattern of these metalloproteases were also performed. Western blot analysis, using an anti-gp63 antibody, revealed polypeptide patterns with a similar profile to that observed in zymographic analysis. Flow cytometry and fluorescence microscopy analyses corroborated the presence of metalloproteases with homologous domains to gp63 in the parasites and revealed differences in the expression level of such molecules among the isolates. The cellular distribution of metalloproteases, assessed by confocal analysis, showed the existence of intracellular metalloproteases with homologous domains to gp63, predominantly located near the flagellar pocket. Finally, it was observed that differential zymographic profiles of metalloproteases exhibited by L. (V.) braziliensis isolates remain unaltered during prolonged in vitro culture, suggesting that the proteolytic activity pattern is a stable phenotypic characteristic of these parasites. PMID- 18423420 TI - Peptides containing T cell epitopes, derived from Sm14, but not from paramyosin, induce a Th1 type of immune response, reduction in liver pathology and partial protection against Schistosoma mansoni infection in mice. AB - Sm14 and paramyosin are two major Schistosoma mansoni vaccine candidate antigens. Recently, we have identified Sm14 and paramyosin epitopes that are recognized by T cells of resistant individuals living in endemic areas for schistosomiasis. Herein, mice were immunized with these peptides separately or in association in order to evaluate their vaccine potential. Immunization of mice with Sm14 peptides alone or mixed with paramyosin peptides was able to induce 26%-36.7% or 28%-29.2% of worm burden reduction, 67% or 46% of intestinal eggs reduction and also 54%-61% or 43%-52% of liver pathology reduction, respectively. Protection was associated with a Th1 type of immune response induced by Sm14 peptide immunization. In contrast, paramyosin peptide vaccination did not engender protective immunity or liver pathology reduction and immunization was associated with a Th2 type of immune response. PMID- 18423421 TI - New treatments of diabetes: the beta-amylin agonists. PMID- 18423422 TI - [Anaplastic thyroid carcinoma]. AB - Anaplastic thyroid carcinoma may represent the ultimate dedifferentiation step of thyroid tumorigenesis and is one of the poorest cancers in human. It accounts for less than 2% of thyroid cancers and affects older patients in their sixth to eighth decade. Usual clinical presentation is a rapidly growing thyroid mass invading surrounding structures with compressive symptoms. Cervical lymph nodes enlargement and distant metastases occur frequently. Though cytological results obtained by fine needle aspiration may be suggestive of diagnosis, tissue biopsy for immunohistochemical study can be necessary to exclude lymphoma and to validate aggressive therapies. Patients developing anaplastic thyroid cancer must be referred urgently in cancer centers to plan multimodality therapeutic approach depending on their performance status. The treatment regimen combines surgery when feasible, hyperfractionated and accelerated external beam radiotherapy and doxorubicin based chemotherapy. Such treatment can provide control of locoregional disease but does not impact on overall survival in patients with distant metastases. The prognosis is dismal with a mean survival of four to nine months after diagnosis. Long survivors are patients with emerging disease presenting a resectable tumor and receiving adjuvant radiotherapy and/or chemotherapy. Therapeutic researches investigate redifferenciation strategies and targeted therapies to inhibit EGF receptors and neoplastic angiogenesis. Primary prevention of this lethal disease may consist of adequate treatment of differentiated thyroid cancers and goiters in elderly. PMID- 18423423 TI - [A proteomic approach of the endocrine tumors]. PMID- 18423424 TI - Decomposing intra-subject variability in children with attention deficit/hyperactivity disorder. AB - BACKGROUND: Increased intra-subject response time standard deviations (RT-SD) discriminate children with attention-deficit/hyperactivity disorder (ADHD) from healthy control subjects. The RT-SD is averaged over time; thus it does not provide information about the temporal structure of RT variability. We previously hypothesized that such increased variability might be related to slow spontaneous fluctuations in brain activity occurring with periods between 15 sec and 40 sec. Here, we investigated whether these slow RT fluctuations add unique differentiating information beyond the global increase in RT-SD. METHODS: We recorded RT at 3-sec intervals for 15 min during an Eriksen flanker task for 29 children with ADHD and 26 age-matched typically developing control subjects (TDC) (mean ages 12.5 +/- 2.4 and 11.6 +/- 2.5; 26 and 12 boys, respectively). The primary outcome was the magnitude of the spectral component in the frequency range between .027 and .073 Hz measured with continuous Morlet wavelet transform. RESULTS: The magnitude of the low-frequency fluctuation was greater for children with ADHD compared with TDC (p = .02, d = .69). After modeling ADHD diagnosis as a function of RT-SD, adding this specific frequency range significantly improved the model fit (p = .03; odds ratio = 2.58). CONCLUSIONS: Fluctuations in low frequency RT variability predict the diagnosis of ADHD beyond the effect associated with global differences in variability. Future studies will examine whether such spectrally specific fluctuations in behavioral responses are linked to intrinsic regional cerebral hemodynamic oscillations that occur at similar frequencies. PMID- 18423426 TI - Genetic variation in the DAOA (G72) gene modulates hippocampal function in subjects at high risk of schizophrenia. AB - BACKGROUND: Strong evidence exists for an association between genetic variation in the gene DAOA (D-amino acid oxidase activator, also known as G72) and risk for schizophrenia. Preliminary evidence in healthy control subjects has implicated genetic variation in the DAOA gene in the modulation of hippocampal complex and prefrontal cortex activation. METHODS: Assessment was performed on 61 subjects at high genetic risk of schizophrenia for familial reasons. All subjects were genotyped for two closely linked single nucleotide polymorphisms in the DAOA gene complex, M23 (rs3918342) and M24 (rs1421292), that have previously shown association with schizophrenia. The effect of genotype on brain activation was assessed with functional magnetic resonance imaging data gathered during performance of the verbal initiation section of the Hayling Sentence Completion Task. RESULTS: Differences between DAOA genotype groups were seen in the activation of the left hippocampus and parahippocampus in the contrast of sentence completion versus rest. In addition the DAOA genotype groups differed in their recruitment of right inferior prefrontal cortex in relation to increasing task difficulty. The effects of genotype on brain activation could not be explained in terms of differences in grey matter density. CONCLUSIONS: These results support the view that genetic variation in the DAOA gene influences hippocampal complex and prefrontal cortex function, an effect that might be particularly prominent in the context of enhanced genetic risk for schizophrenia. PMID- 18423425 TI - Orexin mediates the expression of precipitated morphine withdrawal and concurrent activation of the nucleus accumbens shell. AB - BACKGROUND: The lateral hypothalamic neuropeptide orexin (or hypocretin) is implicated in drug addiction. Although a role for orexin has been shown in reward and dependence, the molecular and neural mechanisms are unclear. We investigated the mechanism and neuroanatomic basis of orexin's role in morphine withdrawal. METHODS: C57BL/6J mice received chronic morphine followed by naloxone (0 or 1 mg/kg, subcutaneous) to precipitate withdrawal. Before naloxone, mice received SB 334867 (0 or 20 mg/kg, intraperitoneal), an orexin 1 receptor (Ox1r) antagonist. Using immunohistochemistry, c-Fos, a marker of cell activation, was quantified in the nucleus accumbens (Acb), lateral hypothalamus (LH), ventral tegmental area (VTA), and locus coeruleus (LC). Retrograde tracing with fluorogold (FG) was performed to determine whether orexin neurons project directly to the Acb. RESULTS: SB-334867 before naloxone significantly attenuated withdrawal symptoms. Withdrawal was accompanied by an increase in c-Fos expression in the Acb shell (AcbSh), which was reduced by SB-334867 but had no effect on the VTA or the LC. Morphine withdrawal increased c-Fos expression in the dorsomedial (DMH) and perifornical (PFA) regions but not in the lateral region of the LH (LLH). Orexin neurons do not appear to form direct connections with Acb neurons. CONCLUSIONS: Altogether, these data demonstrate that orexin, acting via Ox1r, is critical for the expression of morphine withdrawal. AcbSh activation during withdrawal is dependent on Ox1r function and is likely mediated by indirect action of LH orexin neurons. PMID- 18423427 TI - Collateral axonal projections from rostral ventromedial medullary nitric oxide synthase containing neurons to brainstem autonomic sites. AB - The magnocellular reticular nucleus and adjacent lateral paragigantocellular nucleus have been shown to contain a large population of nitric oxide synthase (NOS) immunoreactive neurons. However, little is known about the projections of these neurons within the central nervous system. Retrograde tract-tracing techniques combined with immunohistochemistry were used in this study to investigate whether NOS neurons in this rostral ventromedial medullary (RVMM) region send collateral axonal projections to autonomic sites in the nucleus of the solitary tract (NTS) and in the nucleus ambiguus (Amb). Fluorogold and/or rhodamine labeled latex microspheres were microinjected into the NTS and Amb at sites that elicited bardycardia and/or depressor responses (l-glutamate; 0.25 M; 10 nl). After a survival period of 10-14 days, the rats were sacrificed and tissue sections of the brainstem were processed immunohistochemically for the identification of NOS containing neuronal perikarya. After unilateral injection of the tract-tracers into the NTS and Amb, retrogradely labeled neurons were observed bilaterally throughout the RVMM region. Of the number of RVMM neurons retrogradely labeled from the NTS (684+/-143), 9% were found to be immunoreactive to NOS. Similarly, of those RVMM neurons retrogradely labeled from the Amb (963+/ 207), 7% also contained NOS immunoreactivity. Neurons with collateral axonal projections to NTS and Amb (14% and 10%, respectively) were observed predominantly within a region of RVMM that extended co-extensively with approximately the rostrocaudal extent of the facial nucleus. Of these double labeled neurons, 36.4+/-20 (39%) were also found to be immunoreactive to NOS. These data indicate that the RVMM contains at least three population of NOS neurons that send axons to innervate functionally similar cardiovascular responsive sites in the NTS and Amb. Although the function of these NOS containing medullary pathways in cardiovascular control is not known, it is likely that those with collateral axonal projections represent the anatomical substrate by which the RVMM may simultaneously coordinate cardiovascular responses during physiological changes associated with respiration and/or motor movements. PMID- 18423428 TI - Role of beta1-adrenoceptor in the basolateral amygdala of rats with anxiety-like behavior. AB - There are evidence suggesting that the function of adrenergic receptor is affected in the amygdala of animals with anxiety-like behavior. However, beta adrenoceptor (beta-AR) subtypes, consisting of three subtypes, exert different effects on anxiety regulation. In order to determine the function of the beta1-AR subtype in anxiety-like behavior, we investigated the change of beta1-AR expression by immunostaining in the basolateral amygdala (BLA) of rats treated by conditional fear training. The results indicated that the level of beta1-AR was significantly increased in the BLA of fear-conditioned animals as compared that of controls. In animal behavioral tests, animals treated with selective beta1-AR antagonist metoprolol before conditional fear training exhibited a significant attenuation of anxiety-like behavior characterized by increased percentage of time spent and percentage of entries in the open arms, and increased number of head-dips in the elevated plus-maze (EPM) test compared with the animals treated with only saline. Furthermore, the rats pretreated with metoprolol in the conditional fear training significantly decreased the freezing behavior in the test compared with the controls. The results suggested that the beta1-AR played an important role in anxiety-like behavior, and inhibition of the beta1-AR in the BLA could produce anxiolytic effect. PMID- 18423429 TI - Neurobehavioral basis of the impaired nurturing in mice lacking the immediate early gene FosB. AB - The transcription factor FosB is induced in neurons of the medial preoptic area (MPOA) during parenting, through activation of the extracellular signal-regulated kinase (ERK). FosB mutant (-/-) postpartum mice and virgin mice that are exposed to pups show defective nurturing behavior. The FosB (-/-) MPOA fails to fully up regulate SPRY1 and Rad, the feedback regulators of ERK and calcium signaling, respectively. Here we studied FosB function by examining the gene expression profiles and the behavioral characteristics of FosB (-/-) mice. We found that FosB (-/|-) mice exhibited not only decreased parenting but also decreased infanticide compared with (+/) littermates. We then performed gene expression analysis in the MPOA of FosB (-/-) mice compared with the wild-type littermates. We found up-regulation of glial fibrillary acidic protein (GFAP), C4, and Ela1 mRNA in the MPOA of FosB (-/-) mice; all of these gene products were implicated in general neuropathological conditions. Immunohistochemical analysis showed that up-regulation of GFAP was not restricted to MPOA but extended throughout the forebrain, including the cerebral cortex and striatum. Such pervasive GFAP up regulation suggested that FosB (-/-) mice might have other behavioral abnormalities than nurturing. Indeed, these mice showed a clear alteration in emotionality, detected by the acoustic startle, elevated plus maze, and passive avoidance tests. These results suggest that FosB (-/-) mice have broader neurobehavioral dysfunctions, with which the nurturing defect might share the common mechanism. PMID- 18423431 TI - When facts go down the rabbit hole: contrasting features and objecthood as indexes to memory. AB - People will often look to empty, uninformative locations in the world when trying to recall spoken information. This spatial indexing behaviour occurs when the information had previously been associated with those locations. It remains unclear, however, whether this behaviour is an example of a simple association across perceptual and cognitive systems, or whether location information plays a role in memory retrieval. In the current study, we investigate whether higher level cognitive processes, such as object-based attention, are involved in spatial indexing. Participants saw creatures burrowing around the screen, appearing from underground to tell them facts. They saw the same creature in two locations, or two identical creatures in two locations, depending on spatiotemporal cues conveyed by a burrowing animation. While answering questions, we found that participants relied on these spatiotemporal cues, fixating the previous locations of the creature associated with the relevant fact, rather than the location of an identical creature. We interpret these findings in terms of an object-based attentional mechanism that is common to semantic memory and scene perception, and allows 'external memory' to be exploited in a dynamic environment. PMID- 18423430 TI - A distinct subset of natural killer T cells produces IL-17, contributing to airway infiltration of neutrophils but not to airway hyperreactivity. AB - Activated natural killer T (NKT) cells produce a broad range of cytokines, including IL-4 and IFN-gamma, that determine immunomodulatory functions in various animal models. In this report, we show that a well-known proinflammatory cytokine, IL-17 is also produced by a distinct population of NKT cells upon TCR stimulation. Administration of alpha-galactosylceramide (alpha-GalCer), a strong agonist of NKT cells, induces rapid IL-17 production by a small population of NKT cells, mostly belonging to a population different from that of IL-4- and IFN gamma-producing NKT cells. IL-17-producing NKT cells showed unresponsiveness after stimulation of alpha-GalCer as conventional NKT cells. During airway inflammation induced by pulmonary activation of NKT cells with alpha-GalCer, IL 17 contributes to the infiltration of neutrophils into the airway but has no effect on airway hyperreactivity (AHR). These results indicate that TCR stimulation induces IL-17 expression by a novel population of NKT cells and may help to explain diverse NKT cell functions. PMID- 18423432 TI - Flashbulb memories: special but not iconic. PMID- 18423433 TI - Reviews of popular science books. PMID- 18423434 TI - Cofilin/ADF is required for retinal elongation and morphogenesis of the Drosophila rhabdomere. AB - Drosophila photoreceptors undergo marked changes in their morphology during pupal development. These changes include a five-fold elongation of the retinal cell body and the morphogenesis of the rhabdomere, the light sensing structure of the cell. Here we show that twinstar (tsr), which encodes Drosophila cofilin/ADF (actin-depolymerizing factor), is required for both of these processes. In tsr mutants, the retina is shorter than normal, the result of a lack of retinal elongation. In addition, in a strong tsr mutant, the rhabdomere structure is disorganized and the microvilli are short and occasionally unraveled. In an intermediate tsr mutant, the rhabdomeres are not disorganized but have a wider than normal structure. The adherens junctions connecting photoreceptor cells to each other are also found to be wider than normal. We propose, and provide data supporting, that these wide rhabdomeres and adherens junctions are secondary events caused by the inhibition of retinal elongation. These results provide insight into the functions of the actin cytoskeleton during morphogenesis of the Drosophila eye. PMID- 18423435 TI - Drosophila Naked cuticle (Nkd) engages the nuclear import adaptor Importin-alpha3 to antagonize Wnt/beta-catenin signaling. AB - Precise control of Wnt/beta-catenin signaling is critical for animal development, stem cell renewal, and prevention of disease. In the fruit fly Drosophila melanogaster, the naked cuticle (nkd) gene limits signaling by the Wnt ligand Wingless (Wg) during embryo segmentation. Nkd is an intracellular protein that is composed of separable membrane- and nuclear-localization sequences (NLS) as well as a conserved EF-hand motif that binds the Wnt receptor-associated scaffold protein Dishevelled (Dsh), but the mechanism by which Nkd inhibits Wnt signaling remains a mystery. Here we identify a second NLS in Nkd that is required for full activity and that binds to the canonical nuclear import adaptor Importin-alpha3. The Nkd NLS is similar to the Importin-alpha3-binding NLS in the Drosophila heat shock transcription factor (dHSF), and each Importin-alpha3-binding NLS required intact basic residues in similar positions for nuclear import and protein function. Our results provide further support for the hypothesis that Nkd inhibits nuclear step(s) in Wnt/beta-catenin signaling and broaden our understanding of signaling pathways that engage the nuclear import machinery. PMID- 18423436 TI - Polychaetoid controls patterning by modulating adhesion in the Drosophila pupal retina. AB - Correct cellular patterning is central to tissue morphogenesis, but the role of epithelial junctions in this process is not well-understood. The Drosophila pupal eye provides a sensitive and accessible model for testing the role of junction associated proteins in cells that undergo dynamic and coordinated movements during development. Mutations in polychaetoid (pyd), the Drosophila homologue of Zonula Occludens-1, are characterized by two phenotypes visible in the adult fly: increased sensory bristle number and the formation of a rough eye produced by poorly arranged ommatidia. We found that Pyd was localized to the adherens junction in cells of the developing pupal retina. Reducing Pyd function in the pupal eye resulted in mis-patterning of the interommatidial cells and a failure to consistently switch cone cell contacts from an anterior-posterior to an equatorial-polar orientation. Levels of Roughest, DE-Cadherin and several other adherens junction-associated proteins were increased at the membrane when Pyd protein was reduced. Further, both over-expression and mutations in several junction-associated proteins greatly enhanced the patterning defects caused by reduction of Pyd. Our results suggest that Pyd modulates adherens junction strength and Roughest-mediated preferential cell adhesion. PMID- 18423438 TI - A new method, using cis-regulatory control, for blocking embryonic gene expression. AB - Many genes, and particularly regulatory genes, are utilized multiple times in unrelated phases of development. For studies of gene function during embryogenesis, there is often need of a method for interfering with expression only at a specific developmental time or place. Here we show that in sea urchin embryos cis-regulatory control systems which operate only at specific times and places can be used to drive expression of short designed sequences targeting given primary transcripts, thereby effectively taking out the function of the target genes. The active sequences are designed to be complementary to intronic sequences of the primary transcript of the target genes. In this work, the target genes were the transcription factors alx1 and ets1, both required for skeletogenesis, and the regulatory drivers were from the sm30 and tbr genes. The sm30 gene is expressed only after skeletogenic cell ingression. When its regulatory apparatus was used as driver, the alx1 and ets1 repression constructs had the effect of preventing postgastrular skeletogenesis, while not interfering with earlier alx1 and ets1 function in promoting skeletogenic mesenchyme ingression. In contrast, repression constructs using the tbr driver, which is active in blastula stage, block ingression. This method thus provides the opportunity to study regulatory requirements of skeletogenesis after ingression, and may be similarly useful in many other developmental contexts. PMID- 18423437 TI - Zonula occludens-1 (ZO-1) is involved in morula to blastocyst transformation in the mouse. AB - It is unknown whether or not tight junction formation plays any role in morula to blastocyst transformation that is associated with development of polarized trophoblast cells and fluid accumulation. Tight junctions are a hallmark of polarized epithelial cells and zonula occludens-1 (ZO-1) is a known key regulator of tight junction formation. Here we show that ZO-1 protein is first expressed during compaction of 8-cell embryos. This stage-specific appearance of ZO-1 suggests its participation in morula to blastocyst transition. Consistent with this idea, we demonstrate that ZO-1 siRNA delivery inside the blastomeres of zona weakened embryos using electroporation not only knocks down ZO-1 gene and protein expressions, but also inhibits morula to blastocyst transformation in a concentration-dependent manner. In addition, ZO-1 inactivation reduced the expression of Cdx2 and Oct-4, but not ZO-2 and F-actin. These results provide the first evidence that ZO-1 is involved in blastocyst formation from the morula by regulating accumulation of fluid and differentiation of nonpolar blastomeres to polar trophoblast cells. PMID- 18423439 TI - Role of brain nicotinic acetylcholine receptor in centrally administered corticotropin-releasing factor-induced elevation of plasma corticosterone in rats. AB - The present study was undertaken to clarify the central mechanisms involved in the intracerebroventricularly administered corticotropin-releasing factor-induced elevation of plasma corticosterone in urethane- and alpha-chloralose-anesthetized rats using microdialysis and immunohistochemical techniques. When corticotropin releasing factor was given at 0.5, 1.5, and 3.0 nmol/animal intracerebroventricularly, it dose-dependently increased noradrenaline release but not adrenaline release in the hypothalamic paraventricular nucleus. The 1.5 nmol/animal dose of corticotropin-releasing factor-induced noradrenaline release was attenuated by CP-154,526 (butyl-ethyl-{2,5-dimethyl-7-(2,4,6 trimethylphenyl) 7H-pyrrolo[2,3-d]pyrimidin-4-yl}amine), a selective corticotropin-releasing factor receptor 1 antagonist, at 1.3 micromol/animal, intracerebroventricularly, and was also abolished by phentolamine at 0.66 micromol/animal, intracerebroventricularly. In addition, the corticotropin-releasing factor induced elevation of noradrenaline release in the hypothalamic paraventricular nucleus and plasma corticosterone were abolished by hexamethonium, a non selective nicotinic acetylcholine receptor antagonist, at 1.8 micromol/animal, intracerebroventricularly, and alpha-conotoxin MII, a potent alpha(3)beta(2) nicotinic acetylcholine receptor antagonist, at 30 nmol/animal, i.c.v. Corticotropin-releasing factor at 1.5 nmol/animal, i.c.v. evoked a significant expression of Fos, an immediate-early transcription factor in neurons, on the dopamine-beta-hydroxylase-containing neurons and alpha(3) nicotinic acetylcholine receptor subunit-expressing neurons in the locus coeruleus, but not in the medullary A(1) and A(2) regions containing noradrenergic neurons. These results suggest that centrally administered corticotrophin-releasing factor elevates plasma corticosterone by the corticotropin-releasing factor 1 receptor and alpha(3) subunit-containing nicotinic acetylcholine receptor (probably alpha(3)beta(2) nicotinic acetylcholine receptor) mediated activation of the locus coeruleus noradrenergic neurons projecting to the paraventricular nucleus in rats. PMID- 18423440 TI - Effects of glutamate-related drugs on marble-burying behavior in mice: implications for obsessive-compulsive disorder. AB - Clinical evidence demonstrates altered glutamatergic neurotransmission in patients suffering from obsessive-compulsive disorder (OCD). We examined the effects of glutamate-related drugs on marble-burying behavior, which is an animal model of OCD. The uncompetitive N-methyl-d-aspartate (NMDA) antagonists memantine (10 mg/kg, i.p.) and amantadine (30 mg/kg, i.p.) significantly inhibited marble burying behavior without affecting locomotor activity in mice. Similarly, the uncompetitive NMDA receptor antagonist 5R,10S-(+)-5-methyl-10,11-dihydro-5H dibenzo(a,d)cyclohepten-5,10-imine hydrogen maleate (MK-801, 0.3 mg/kg, i.p.) inhibited marble-burying behavior. However, MK-801 at the same dose markedly increased locomotor activity. By contrast, the alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionate (AMPA) receptor antagonist 1,2,3,4-tetrahydro-6-nitro-2,3 dioxo-benzo[f]quinoxaline-7-sulfonamide (NBQX) and the glutamate release inhibitor riluzole showed no effect on marble-burying behavior and significant suppression of locomotor activity. MK-801 (0.3 mg/kg, i.p.) and memantine (10 mg/kg, i.p.) significantly disrupted prepulse inhibition as an operational measure of sensorimotor gating. By contrast, amantadine (30 mg/kg, i.p.) did not affect prepulse inhibition. These findings suggest that amantadine could be a useful drug for the treatment of OCD. PMID- 18423441 TI - Effect of co-administration of the selective 5-HT1A receptor antagonist WAY 100,635 and selective 5-HT1B/1D receptor antagonist GR 127,935 on anxiolytic effect of citalopram in conditioned fear stress in the rat. AB - This study investigated the effect of co-administration of the selective 5-HT1A receptor antagonist WAY 100,635 and selective 5-HT(1B/1D) receptor antagonist GR 127,935 with a subactive dose of citalopram [selective serotonin (5-HT) reuptake inhibitor (SSRI)] on the expression of conditioned freezing, an index of fear. In the present study, acute administration of citalopram (s.c.) reduced freezing significantly at high doses (10, 30 and 100 mg/kg), while showing no significant effect at low doses (1 and 3 mg/kg). Co-administration of WAY 100,635 (0.15 mg/kg) with citalopram (3 mg/kg) reduced freezing markedly and significantly, as compared with either drug alone. However, the addition of GR 127,935 (4 mg/kg) did not potentiate the effects of citalopram (3 mg/kg) on freezing and did not enhance the effect of WAY 100,635 (0.15 mg/kg) with citalopram (3 mg/kg). Co administration of WAY 100,635 (0.15 mg/kg) or GR 127,935 (4 mg/kg) gave no effect on high-dose citalopram (30 mg/kg)-induced inhibition of freezing behavior. These results suggest that co-administration of WAY 100,635 (0.15 mg/kg) strengthens the anxiolytic effect of citalopram (3 mg/kg) by facilitating central 5-HT neurotransmission. Since GR 127,935 (4 mg/kg) failed to accelerate the inhibition of freezing induced by citalopram (3 mg/kg) with WAY 100,635 (0.15 mg/kg) or citalopram (3 mg/kg) alone, it is suggested that blocking 5-HT1A receptors is more effective in facilitating the anxiolytic effect of citalopram than blocking 5-HT1B/1D receptors. PMID- 18423442 TI - The use of the hippocampal slice preparation in the study of Alzheimer's disease. AB - In the present article we show how studying synaptic mechanisms in hippocampal slice preparations provides information that may be useful in, firstly, the understanding of the aetiology of Alzheimer's disease and, secondly, in the development of novel therapies for dementia. We use several examples, drawn from our own work: (i) The identification of the function of AMPA receptors and NMDA receptors in synaptic transmission and synaptic plasticity. (ii) The discovery of mechanisms that can regulate the activation of NMDA receptors. (iii) The use of transgenic models of Alzheimer's disease. (iv) The identification of a mechanism that can account for the cognitive enhancing effects of the NMDA receptor antagonist memantine. (v) The discovery of a role of glycogen synthase kinase 3beta (tau kinase) in synaptic plasticity. PMID- 18423443 TI - Corticosteroid receptor-gene variants: modulators of the stress-response and implications for mental health. AB - The stress-response, including autonomic and hypothalamic-pituitary-adrenal (HPA) axis reactivity, is essential for maintaining homeostasis during a challenge. Brain mineralocorticoid receptors and glucocorticoid receptors operate in balance to coordinate the stress-response. Genetic variants in both the human mineralocorticoid and glucocorticoid receptor-genes have been functionally characterized. In vitro effects of these genetic variants on transactivation and mRNA stability have been described. In vivo, two mineralocorticoid receptor-gene SNPs (-2 G/C (allele frequency: 50%), MR I180V (11%)) and four glucocorticoid receptor-gene SNPs (ER22/23EK (3%), N363S (4%), BclI (37%), A3669G (15%)) are associated with changes in hypothalamic-pituitary-adrenal (HPA) axis reactivity. Importantly, the two mineralocorticoid receptor-gene variants (but none of the glucocorticoid receptor-gene variants) also associate with changes in autonomic output as measured as increased heart beat following a psychosocial stress (TSST). Moreover, several of these mineralocorticorticoid receptor- and glucocorticoid receptor variants have been found associated with stress-related disorders, including depression. These data indicate that dysregulation of mineralocorticoid- and glucocorticoid receptor are causative in the pathogenesis of depression. Moreover, these mineralocorticoid- and glucocorticoid receptor gene variants constitute part of the genetic make up that determines individual stress-responsiveness inducing vulnerability to disease. Furthermore, mineralocorticoid- and glucocorticoid receptors are drug targets, thereby aiming at the underlying mechanisms of stress-related disorders. PMID- 18423444 TI - Translational research into sexual disorders: pharmacology and genomics. AB - The existence of sexual dysfunctions in men, including premature and retarded ejaculation poses challenges to develop translational models in rats that may help in improving treatment and delineate the neural mechanisms of action. Most of our current understanding of the neurobiology, neuroanatomy and psychopharmacology of sexual behavior and ejaculatory function has been derived from preclinical studies in the rat. When large populations of male rats are tested on sexual activity during four successive tests, over time individual rats display a very stable sexual behavior that is either slow, normal or fast as characterized by the number of ejaculations performed. These sexual endophenotypes are postulated as rat counterparts of premature (fast rats) or retarded ejaculation (slow rats). Psychopharmacology in these endophenotypes may help to delineate the underlying mechanisms and pathology. This is illustrated by the effects of serotonergic antidepressants and serotonergic compounds on sexual and ejaculatory behavior of rats. Further unravelling of sexual endophenotypes may benefit from the use of chromosomal substitution strains in mice that enable the localization of relevant chromosomal areas and genes involved in ejaculation processes. These preclinical studies and models contribute to a better understanding of the neurobiology of ejaculation and boost the development of novel drug targets to treat ejaculatory disorders such as premature and retarded ejaculation. PMID- 18423446 TI - A life course of adiposity and dementia. AB - Adiposity, commonly measured as body mass index (BMI), may influence or be influenced by brain structures and functions involved in dementia processes. Adipose tissue changes in degree and intensity over the lifespan, and has been shown to influence brain development in relationship to early and late measures of cognitive function, intelligence, and disorders of cognition such as dementia. A lower BMI is associated with prevalent dementia, potentially due to underlying brain pathologies and correspondingly greater rates of BMI or weight decline observed during the years immediately preceding clinical dementia onset. However, high BMI during mid-life or at least approximately 5-10 years preceding clinical dementia onset may increase risk. The interplay of adiposity and the brain occurring over the course of the lifespan will be discussed in relationship to developmental origins, mid-life sequelae, disruptions in brain structure and function, and late-life changes in cognition and dementia. Characterizing the life course of adiposity among those who do and do not become demented enhances understanding of biological underpinnings relevant for understanding the etiologies of both dementia and obesity and their co-existence. PMID- 18423447 TI - Neurovascular pharmacology of migraine. AB - Migraine is a paroxysmal neurovascular disorder, which affects a significant proportion of the population. Since dilation of cranial blood vessels is likely to be responsible for the headache experienced in migraine, many experimental models for the study of migraine have focussed on this feature. The current review discusses a model that is based on the constriction of carotid arteriovenous anastomoses in anaesthetized pigs, which has during the last decades proven of great value in identifying potential antimigraine drugs acting via a vascular mechanism. Further, the use of human isolated blood vessels in migraine research is discussed. Thirdly, we describe an integrated neurovascular model, where dural vasodilatation in response to trigeminal perivascular nerve stimulation can be studied. Such a model not only allows an in-depth characterization of directly vascularly acting drugs, but also of drugs that are supposed to act via inhibition of vasodilator responses to endogenous neuropeptides, or of drugs that inhibit the release of these neuropeptides. We discuss the use of this model in a study on the influence of female sex hormones on migraine. Finally, the implementation of this model in mice is considered. Such a murine model allows the use of genetically modified animals, which will lead to a better understanding of the ion channel mutations that are found in migraine patients. PMID- 18423448 TI - Membrane-type 1 matrix metalloproteinase regulates cell migration during zebrafish gastrulation: evidence for an interaction with non-canonical Wnt signaling. AB - Key to invasiveness is the ability of tumor cells to modify the extracellular matrix, become motile, and engage in directed migration towards the vasculature. One significant protein associated with metastatic progression is membrane-type 1 matrix metalloproteinase (MT1-MMP/MMP14). How MMP14 activity is coordinated with other signaling pathways to regulate cell migration in vivo is largely unknown. Here we have used zebrafish embryogenesis as a model to understand the potential relationship between MMP14-dependent pericellular proteolysis, cell polarity, and motility. Knockdown of zebrafish Mmp14 function disrupted gastrulation convergence and extension cell movements and craniofacial morphogenesis. Using time-lapse imaging and morphometric analyses, we show that Mmp14 is required for proper cell polarity underlying the directed migration of mesodermal cells during gastrulation. We have identified a genetic interaction between mmp14 and non canonical Wnt signaling, a pathway that also regulates cell polarity in embryonic tissues and is increasingly being linked with tumor cell migration. Finally, we demonstrate that Van Gogh-like 2, a key regulator of the non-canonical Wnt pathway, co-localizes with MMP14 and becomes redistributed towards the leading edge of polarized human cancer cells. Together, our results support the notion that pathways regulating pericellular proteolysis and cell polarity converge to promote efficient cell migration. PMID- 18423449 TI - Anterior segment dysgenesis in the eyes of mice deficient for the high-mobility group transcription factor Sox11. AB - We describe that Sox11, a member of the group C of the Sox transcription factor family, is critically required during the morphogenetic processes of early eye development, and that lack of Sox11 results in ocular anterior segment dysgenesis (ASD). Sox11-deficient mice show a persistent lens stalk, a delay in lens formation, and the phenotypes of Peters' anomaly and microphthalmia at birth. In addition, the optic fissure does not close in the anterior halves of the eyes resulting in anterior coloboma. The delay in lens formation is associated with a reduced mitotic activity in the lens placode during its invagination into the optic cup. No changes in Pax6 expression are observed in the developing eyes of Sox11-/- mice, whereas the expression of Sox11 is reduced in optic cup, optic vesicle and lens placode of Pax6+/- embryos and in the optic vesicle of Pax6-/- mice. Transfection experiments show an increase in Sox11 expression when higher doses of Pax6 are present. Considerably smaller amounts of BMP7 are expressed in lens and optic cup of Sox11-/- mice as compared to their wild-type littermates. We conclude that Sox11 is required during separation of the lens vesicle from the surface ectoderm and the closure of the anterior optic fissure. The expression of Sox11 in early eye development is under control of Pax6, and changes in BMP7 signalling appear to be involved in the effects of Sox11 on anterior eye development. Our findings suggest that SOX11 might similarly be involved in the pathogenesis of ASD in human patients. PMID- 18423450 TI - The urokinase system of plasminogen activator plays a role in amyotrophic lateral sclerosis (ALS) pathogenesis. PMID- 18423451 TI - A dopamine receptor antagonist L-745,870 suppresses microglia activation in spinal cord and mitigates the progression in ALS model mice. AB - Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease characterized by a selective loss of motor neurons in the motor cortex, brainstem, and spinal cord. It has been shown that oxidative stress plays a pivotal role in the progression of this motor neuron loss. We have previously reported that L 745,870, a dopamine D4 receptor antagonist, selectively inhibits oxidative stress induced cell death in vitro and exerts a potent neuroprotective effect against ischemia-induced neural cell damage in gerbil. To investigate the efficacy of L 745,870 in the treatment of ALS, we here conducted a chronic administration of L 745,870 to transgenic mice expressing a mutated form of human superoxide dismutase gene (SOD1(H46R)); a mouse model of familial ALS, and assessed whether the mice benefit from this treatment. The pre-onset administration of L-745,870 significantly delayed the onset of motor deficits, slowed the disease progression, and extended a life span in transgenic mice. These animals showed a delayed loss of anterior horn cells in the spinal cord concomitant with a reduced level of microglial activation at a late symptomatic stage. Further, the post onset administration of L-745,870 to the SOD1(H46R) transgenic mice remarkably slowed the disease progression and extended their life spans. Taken together, our findings in a rodent model of ALS may have implication that L-745,870 is a possible novel therapeutic means to the treatment of ALS. PMID- 18423452 TI - Rectosigmoid endometriosis with unusual presentation at magnetic resonance imaging. AB - We report on a patient with a bowel endometriotic nodule that was hypointense on both T1- and T2-weighted magnetic resonance imaging (MRI); histologic examination revealed that the MRI appearance of the nodule was due to the presence of extensive fibrosis. Bowel endometriotic nodules with extensive fibrosis and limited endometrial tissue have a signal intensity similar to that of the muscle on both T1- and T2-weighted images. In these patients, additional techniques such as multislice computed tomography enteroclysis may be used for adequate diagnosis before surgery. PMID- 18423453 TI - Safety and efficacy of a single-rod etonogestrel implant (Implanon): results from 11 international clinical trials. AB - OBJECTIVE: To present efficacy, safety, and bleeding profile results from the clinical trials that supported the U.S. Food and Drug Administration filing for the approval of a single-rod etonogestrel (ENG) contraceptive implant (Implanon). DESIGN: Integrated analysis of 11 international clinical trials. SETTING: Contraceptive clinics in U.S., Chile, Asia, and Europe. PATIENT(S): A total of 942 healthy women, aged 18 to 40 years. INTERVENTION(S): Insertion of an ENG implant. Most women were enrolled in studies lasting either 2 or 3 years. MAIN OUTCOMES MEASURE(S): Efficacy was measured by the cumulative Pearl Index in women .01). LIMITATIONS: A single center study and small sample. CONCLUSIONS: Endoscopic thermal ablation with APC is effective in managing UGIB and in reducing transfusion requirements in patients admitted for GI hemorrhage because of different endoscopic types of GVE. PMID- 18423467 TI - Actinomyces-like organisms from a vaginal granuloma following intravaginal slingplasty with polypropylene mesh. PMID- 18423468 TI - Angiogenic factors for the prediction of pre-eclampsia in women with abnormal midtrimester uterine artery Doppler velocimetry. AB - OBJECTIVE: To determine whether assessing uterine perfusion and angiogenic factors concurrently in the second trimester improves the prediction of pre eclampsia and intrauterine growth restriction (IUGR). METHOD: Plasma levels of soluble fms-like tyrosine kinase 1 (sFlt-1) and placental growth factor (PlGF) were measured for 108 women with abnormal uterine perfusion on Doppler velocimetry in the 23rd week. Later, 33 cases of pre-eclampsia and 9 of IUGR developed. RESULTS: Compared with the plasma levels of the women whose pregnancies remained normal, sFlt-1 levels were significantly higher and PlGF levels significantly lower in the women whose pregnancies became complicated by pre-eclampsia and/or IUGR (P<0.001). The alterations were more pronounced in cases of early-onset pre-eclampsia and in cases of IUGR that necessitated delivery before 34 weeks. Using the sFlt-1/PlGF ratio, these complications could be predicted with 98% sensitivity, 95% specificity, and 93% positive predictive value. CONCLUSION: Measuring uterine perfusion and angiogenic factors concurrently in the second trimester improved the prediction of pre-eclampsia and IUGR. PMID- 18423469 TI - Anencephaly and organ transplantation. PMID- 18423470 TI - Preoperative evaluation of pelvic masses with combined 18F-fluorodeoxyglucose positron emission tomography and computed tomography. AB - OBJECTIVE: To prospectively evaluate the diagnostic value of combined 18F fluorodeoxyglucose position emission tomography and computed tomography (FDG PET/CT) to discriminate malignant or borderline malignant tumors from benign pelvic masses. METHODS: A prospective study of 30 women with suspected ovarian cancer who presented from July 2006 through August 2007. Selection was based on evidence from ultrasound, magnetic resonance imaging, and rising tumor marker levels. All patients underwent FDG-PET/CT prior to standard debulking surgery for a pelvic mass. RESULTS: The sensitivity and specificity of FDG-PET/CT to detect malignant or borderline malignant pelvic tumors were 71.4% and 81.3%, respectively. The sensitivity and specificity of FDG-PET/CT to detect ovarian cancer were 100% and 85.0%, respectively. The maximum standardized uptake value in borderline tumors was significantly lower compared with malignant tumors, but not significantly different compared with benign tumors. CONCLUSION: FDG-PET/CT had a high diagnostic value in differentiating between malignant and benign tumors, and a low diagnostic value in differentiating between borderline malignant and benign tumors. PMID- 18423472 TI - Measurement of stride parameters using a wearable GPS and inertial measurement unit. AB - Both GPS and inertial measurement units (IMUs) have been extensively used in biomechanical studies. Expensive high accuracy GPS units can provide information about intrastride speed and position, but their application is limited by their size and cost. Single and double integration of acceleration from IMU provides information about short-term fluctuations in speed and position, but suffers from integration error over a longer period of time. The integration of GPS and IMU has been widely used in large and expensive units designed for survey and vehicle navigation. Here we propose a data fusion scheme, which is a Kalman filter based complementary filter and enhances the frequency response of the GPS and IMU used alone. We also report the design of a small (28 g) low cost GPS/IMU unit. Its accuracy after post-processing with the proposed data fusion scheme for determining average speed and intrastride variation was compared to a traditional high cost survey GPS. The low cost unit achieved an accuracy of 0.15 ms(-1) (s.d.) for horizontal speed in cycling and human running across a speed range of 3-10 ms(-1). The stride frequency and vertical displacement calculated based on measurements from the low cost GPS/IMU units had an s.d. of 0.08 Hz and 0.02 m respectively, compared to measurements from high performance OEM4 GPS units. PMID- 18423471 TI - Familial predisposition to cardiovascular risk and disease contributes to cardiovascular risk and disease interacting with other cardiovascular risk factors in diabetes: implication for common soil (JDDM 14). AB - In diabetic population cardiovascular morbidity is high and the effects of genetic predisposition remain elucidated. In a large-scale multicenter-based diabetic population, clinical parameters including conventional cardiovascular risk factors and first-degree family history (FH) of diabetes, hypertension, coronary heart disease (CHD) and stroke were investigated in association with presence of CHD and stroke. Among 3611 diabetic patients, 181 (5.0%) had CHD and 118 (3.3%) had stroke. After adjustment for conventional risk factors, FH of CHD (OR 2.32, p<0.0001) and of diabetes (OR 1.44, p<0.05) were associated with CHD, and FH of stroke (OR 1.86, p<0.01) was associated with stroke. FH of hypertension was significantly associated with presence of hypertension and obesity. Synergistic effect of FH of CHD in combination with hypertension or aging on increasing CHD, and that of FH of stroke in combination with microalbuminuria on increasing stroke were found. FH of diabetes, of hypertension, of CHD and of stroke were significantly associated with FH of each disease, indicating clustering of FH. In diabetic population, FH of CHD and FH of stroke doubled the risk of CHD and stroke, respectively, and had synergistic effect in combination with other risk factors. Clustering of FH may indicate interrelation of genetic predisposition. PMID- 18423473 TI - Indirect determination of trabecular bone effective tissue failure properties using micro-finite element simulations. AB - Trabecular bone strength is marked not only by the onset of local yielding, but also by post-yield behavior. To study and predict trabecular bone elastic and yield properties, micro-finite element (micro-FE) models were successfully applied. However, trabecular bone strength predictions require micro-FE models incorporating post-yield behavior of trabecular bone tissue. Due to experimental difficulties, such data is currently not available. Here we used micro-FE modeling to determine failure behavior of trabecular bone tissue indirectly, by iteratively fitting FE simulation to experimental results. Failure parameters were fitted to an isotropic plasticity model based on Hill's yield function, using materially and geometrically nonlinear micro-FE models of seven bovine trabecular bone specimens. The predictive value of the averaged effective tissue properties was subsequently tested. The results showed that compression softening had to be included on the tissue level in order to accurately describe the apparent-level behavior of the bone specimens. A sensitivity study revealed that the simulated response was less sensitive to variations in the post-yield properties of the bone tissue than variations in the elastic and yield properties. Due to fitting of the tissue properties, apparent-level behavior could be accurately reproduced for each specimen separately. Predictions based on the averaged and fixed tissue properties were less accurate, due to inter specimen variations in the tissue properties. PMID- 18423474 TI - Use of supercritical fluid extraction and gas chromatography-mass spectrometry to obtain amino acid profiles from several genetically modified varieties of maize and soybean. AB - A method using supercritical fluid extraction (SFE) and gas chromatography-mass spectrometry (GC/MS) for obtaining the amino acid profiles of genetically modified maize and soybean is proposed. SFE is carried out in a homemade modular system where amino acids are extracted with carbon dioxide modified with 35% of methanol, in conditions optimized by a central composite design. Once extracted, the amino acids are determined by GC/MS. The method has been applied to three samples of maize derived from MON810, other from NK 603 and a Roundup ready soybean sample. The profiles are compared with those obtained from their corresponding isogenic non-transgenic varieties. Although differences are directly observed in some cases by visual comparison of the chromatograms, the application of ANOVA shows more significant differences. In general terms, isogenic varieties seem to have higher content of several amino acids. PMID- 18423475 TI - Application of chaotropic effect in reversed-phase liquid chromatography of structurally related phenothiazine and thioxanthene derivatives. AB - Chromatographic behavior of two main classes of neuroleptics, derivatives of phenothiazine and thioxanthene in RP systems modified by anionic additives: sodium perchlorate and sodium hexafluorophosphate possessing chaotropic properties, was examined. Influence of the method of pH lowering (by addition of acids: trifluoroacetic or perchloric or by adding the appropriate concentration of phosphate buffer) and the kind of organic modifier in the mobile phase (methanol, acetonitrile) were estimated. Stability of complexes created between protonated drugs and anions of added salts was evaluated by comparison of their desolvation parameters (K), limiting retention factors for unsolved molecules calculated on the basis of chromatographic data. Experimentally obtained parameters were used in QSAR studies. It appeared that chosen parameters reflect not only physico-chemical properties of analytes but also contain information about the strength of their antipsychotic activity. Multidimensional cluster analysis has been performed. On the basis of the results obtained, it could be concluded that chaotropic systems can generate useful parameters for further QSAR studies. PMID- 18423476 TI - Determination of quaternary ammonium herbicides in soils. Comparison of digestion, shaking and microwave-assisted extractions. AB - Very challenging analytical problems arise from the continuous introduction in agriculture of chemical pesticides. Particularly, diquat (DQ), paraquat (PQ) and difenzoquat (DF) are a difficult group of quaternary ammonium herbicides to analyze. This article reviews and addresses the most relevant analytical methods for determining the selected herbicides in soil. We discuss and critically evaluate procedures, such as digestion-based methods, shaking extraction and microwave-assisted extraction (MAE). Clean-up of extracts was performed by solid phase extraction (SPE) using silica cartridges. Detection of these herbicides was carried out by liquid chromatography (LC) coupled to UV detection and mass spectrometry (MS) as confirmatory technique. Recoveries ranged from 98% to 100% by digestion, from no recovered to 61% by shaking, and from 102% to 109% by MAE with estimated quantification limits between 1.0 microg/kg and 2.0 microg/kg by digestion and 5.0 mug/kg and 7.5 microg/kg by MAE using LC/MS-MS as detection technique. The recoveries obtained under the optimum conditions are compared and discussed with those obtained from digestion extraction and MAE. PMID- 18423477 TI - Textural characterization of native and n-alky-bonded silica monoliths by mercury intrusion/extrusion, inverse size exclusion chromatography and nitrogen adsorption. AB - Native and n-alkyl-bonded (n-octadecyl) monolithic silica rods with mesopores in the range between 10 and 25 nm and macropores in the range between 1.8 and 6.0 microm were examined by mercury intrusion/extrusion, inverse size exclusion chromatography (ISEC) and nitrogen sorption. Our results reveal very good agreement for the mesopore size distribution obtained from nitrogen adsorption (in combination with an advanced NLDFT analysis) and ISEC. Our studies highlight the importance of mercury porosimetry for the assessment of the macropore size distribution and show that mercury porosimetry is the only method which allows obtaining a combined and comprehensive structural characterization of macroporous/mesoporous silica monoliths. Our data clearly confirm that mercury porosimetry hysteresis and entrapment have different origin, and indicate the intrinsic nature of mercury porosimetry hysteresis in these silica monoliths. Within this context some silica monoliths show the remarkable result of no entrapment of mercury after extrusion from the mesopore system (i.e. for the first intrusion/extrusion cycle). The results of a systematic study of the mercury intrusion/extrusion behavior into native silica monoliths and monoliths with bonded n-alkyl groups reveals that the macro (through) pore structure, which controls the mass transfer to and from the mesopores, here mainly controls the entrapment behavior. Our data suggest that mercury intrusion/extrusion porosimetry does not only allow to obtain a comprehensive pore structure analysis, but can also serve as a tool to estimate the mass transport properties of silica monoliths to be employed in liquid-phase separation processes. PMID- 18423478 TI - Copper and arsenate co-sorption at the mineral-water interfaces of goethite and jarosite. AB - The co-sorption reaction products of arsenate (As(V)) and copper (Cu(II)) on goethite (alpha-FeOOH) and natro-jarosite (Na(3)Fe(3)(SO(4))(2)(OH)(6)) were investigated with extended X-ray absorption fine structure (EXAFS) spectroscopy to determine if Cu(II) and As(V) would form precipitates or compete with each other for surface sites. The reaction products were prepared by mixing 250 microM Cu(SO(4)) with 10, 25, or 50 microM Na(2)HAsO(4) at pH 5.65 and allowing the mixture to react in 10 m(2) L(-1) goethite or jarosite suspensions for 12 days. In addition, EXAFS data of Cu(SO(4)) and As(V) sorbed on goethite and jarosite were collected as control species. All reaction conditions were under-saturated with respect to common copper bearing minerals: tenorite (CuO), brochantite (Cu(4)(OH)(6)SO(4)), and hydrated clinoclase (Cu(3)(AsO(4))(2)2H(2)O). The extents of the As(V) and Cu(II) surface adsorption reactions showed a strong competitive effect from Cu(II) on As(V) adsorption for a nominal Cu:As mole-ratio of 25:1. With increasing nominal As(V) concentration, As(V) sorption on goethite and jarosite increased without diminishing the amount of Cu(II) sorption. In the absence of either co-sorbate, As(V) and Cu(II) formed the expected surface adsorption species, i.e., bidentate binuclear and edge-sharing surface complexes, consistent with previously published results. In each other's presence, the local bonding environments of As(V) and Cu(II) showed that the co-sorbates form a precipitate on the goethite and jarosite surface at nominal concentrations of 10:1 and 5:1. At nominal Cu:As mole-ratios of 25:1, Cu(II) did not form significantly different surface complexes on goethite or jarosite from those in the absence of As(V), however, As K-edge EXAFS results distinctly showed Cu(II) atoms in As(V)'s local bonding environment on the goethite surface. The structures of the two precipitates were different and depended on the anion-layer structure and possibly the presence of structural oxyanions in the case of jarosite. On goethite, the copper-arsenate precipitate was similar to hydrated clinoclase, while on jarosite, a euchroite-like precipitate (Cu(2)[AsO(4)](OH)3H(2)O, P 2(1)2(1)2(1)) had formed. Despite under-saturated solution conditions, the formation of these precipitates may have occurred due to a seed-formation effect from densely surface adsorbed Cu(II) and As(V) for which the "new" saturation index was significantly lower than homogeneous values would otherwise suggest. Synergistic reactions between two co-sorbates of fundamentally different surface adsorption behaviour can thus be achieved if the number of available sites for surface adsorption is limited. PMID- 18423479 TI - Microscopic approach for the identification of cationic membrane fouling during cheddar cheese whey electroacidification. AB - This is the first time that fouling of cation-exchange membranes during cheddar cheese whey electroacidification with bipolar membranes is reported. A mineral fouling was observed only on the cationic membrane side in contact with the base. The deposit was identified as magnesium hydroxide and this fouling was more important on the cation-exchange membrane situated close to the cathode. Little deposit was formed after six electroacidification runs, but on long time, the buildup of fouling film would lead after many electroacidifications to an important decrease of the system efficiency. Since, fouling of permselective membranes represents one of the major issues in electrodialytic processes, this result will be the basis for the determination of cleaning conditions allowing the prevention of such a fouling. PMID- 18423480 TI - Increased exosome production from tumour cell cultures using the Integra CELLine Culture System. AB - Exosomes are nanometer-sized vesicles, secreted from most cell types, with documented immune-modulatory functions. Exosomes can be purified from cultured cells but to do so effectively, requires maintenance of cells at high density in order to obtain sufficient accumulation of exosomes in the culture medium, prior to purification. Whilst high density cultures can be achieved with cells in suspension, this remains difficult with adherent cells, resulting in low quantity of exosomes for subsequent study. We have used the Integra CELLine culture system, originally designed for hybridoma cultures, to achieve a significant increase in obtainable exosomes from adherent and non-adherent tumour cells. Traditional cultures of mesothelioma cells (cultured in 75 cm(2) flasks) gave an average yield of 0.78 microg+/-0.14 microg exosome/ml of conditioned medium. The CELLine Adhere 1000 (CLAD1000) flask, housing the same cell line, increased exosome yield approximately 12 fold to 10.06 microg+/-0.97 microg/ml. The morphology, phenotype and immune function of these exosomes were compared, and found to be identical in all respects. Similarly an 8 fold increase in exosome production was obtained from NKL cells (a suspension cell line) using a CELLine 1000 (CL1000) flask. The CELLine system also incurred ~5.5 fold less cost and reduced labour for cell maintenance. This simple culture system is a cost effective, useful method for significantly increasing the quantity of exosomes available from cultured cells, without detrimental effects. This tool should prove advantageous in future studies of exosome-immune modulation in cancer and other settings. PMID- 18423481 TI - A novel approach for the generation of human dendritic cells from blood monocytes in the absence of exogenous factors. AB - Human dendritic cells (DCs) for research and clinical applications are typically derived from purified blood monocytes that are cultured in a cocktail of cytokines for a week or more. Because it has been suggested that these cytokine derived DCs may be deficient in some important immunological functions and might not accurately represent antigen presenting cell (APC) populations found under normal conditions in vivo, there is an interest in developing methods that permit the derivation of DCs in a more physiologically relevant manner in vitro. Here, we describe a simple and reliable technique for generating large numbers of highly purified DCs that is based on a one-way migration of blood monocytes through a layer of human umbilical vein endothelial cells (HUVECs) that are cultured to confluency in the upper chamber of a Transwell device. The resultant APCs, harvested from the lower Transwell chamber, resemble other cultured DC populations in their expression of major histocompatibility (MHC) and costimulatory molecules, ability to phagocytose protein antigens and capacity to trigger primary antigen-specific T cell responses. This technique offers several advantages over the standard method of in vitro cytokine-driven DC development, including: (1) the rapidity of this approach, as DC differentiation occurs in only 2 days, (2) the differentiation process itself, which is more akin to the development of DCs under physiologic conditions and (3) the cost-effectiveness of the system, since no monocyte pre-selection is required and DC development occurs in the absence of expensive recombinant cytokines. PMID- 18423482 TI - Evolution of entomopathogenicity in fungi. AB - The recent completions of publications presenting the results of a comprehensive study on the fungal phylogeny and a new classification reflecting that phylogeny form a new basis to examine questions about the origins and evolutionary implications of such major habits among fungi as the use of living arthropods or other invertebrates as the main source of nutrients. Because entomopathogenicity appears to have arisen or, indeed, have lost multiple times in many independent lines of fungal evolution, some of the factors that might either define or enable entomopathogenicity are examined. The constant proximity of populations of potential new hosts seem to have been a factor encouraging the acquisition or loss of entomopathogenicity by a very diverse range of fungi, particularly when involving gregarious and immobile host populations of scales, aphids, and cicadas (all in Hemiptera). An underlying theme within the vast complex of pathogenic and parasitic ascomycetes in the Clavicipitaceae (Hypocreales) affecting plants and insects seems to be for interkingdom host-jumping by these fungi from plants to arthropods and then back to the plant or on to fungal hosts. Some genera of Entomophthorales suggest that the associations between fungal pathogens and their insect hosts appear to be shifting away from pathogenicity and towards nonlethal parasitism. PMID- 18423483 TI - Potential of Lecanicillium spp. for management of insects, nematodes and plant diseases. AB - Fungi in the genus Lecanicillium (formerly classified as the single species Verticillium lecanii) are important pathogens of insects and some have been developed as commercial biopesticides. Some isolates are also active against phytoparasitic nematodes or fungi. Lecanicillium spp. use both mechanical forces and hydrolytic enzymes to directly penetrate the insect integument and the cell wall of the fungal plant pathogen. In addition to mycoparasitism of the plant pathogen, the mode of action is linked to colonization of host plant tissues, triggering an induced systemic resistance. Recently it was demonstrated that development of Lecanicillium hybrids through protoplast fusion may result in strains that inherit parental attributes, thereby allowing development of hybrid strains with broader host range and other increased benefits, such as increased viability. Such hybrids have demonstrated increased virulence against aphids, whiteflies and the soybean cyst nematode. Three naturally occurring species of Lecanicillium, L. attenuatum, L. longisporum, and an isolate that could not be linked to any presently described species based on rDNA sequences have been shown to have potential to control aphids as well as suppress the growth and spore production of Sphaerotheca fuliginea, the causal agent of cucumber powdery mildew. These results suggest that strains of Lecanicillium spp. may have potential for development as a single microbial control agent effective against several plant diseases, pest insects and plant parasitic nematodes due to its antagonistic, parasitic and disease resistance inducing characteristics. However, to our knowledge, no Lecanicillium spp. have been developed for control of phytopathogens or phytoparasitic nematodes. PMID- 18423484 TI - Perkinsus marinus in pleasure oyster Crassostrea corteziensis from Nayarit, Pacific coast of Mexico. AB - Culture of the pleasure oyster Crassostrea corteziensis is emerging as an alternative to the Pacific oyster (Crassostrea gigas) for oyster producers, who face severe mortalities since 1997 in Northwest Mexico. For determining the health status of this species, we conducted a histopathological analysis of cultured populations from two estuaries in the Pacific coast of Mexico. Macroscopical analysis revealed animals with transparent and retracted mantle. Histopathological analysis of these specimens showed tissue alterations and parasitic forms consistent with Perkinsus sp. infection. Stages of the parasite identified included tomont and trophozoites with an eccentric vacuole characteristic of Perkinsus spp. Pieces of tissues of infected oysters were incubated in Fluid Thioglycollate Medium (FTM) resulting in blue-black hypnospores after incubation. The identity of the parasite was confirmed by species specific PCR-based assay in DNA samples from oysters, tissue fractions from FTM cultures, and deparaffined samples with Perkinsus-like parasite detected by histology. Sequencing of positive amplified fragments (307bp) showed a sequence similar to Perkinsus marinus strain TXsc NTS ribosomal RNA gene (100% coverage and 98% identity, GenBank Accession No. AF497479.1) and to P. marinus, Genomic DNA, (100% coverage and 97% identity, GenBank Accession No. S78416.1). The prevalence of P. marinus varied from 1 to 5% in Boca del Camichin and from 1 to 6% in Pozo Chino. In general, the intensity of infection was moderate. The infection was observed in oysters from 31 to 110mm of shell length. This is the first record of P. marinus in oysters from the North America Pacific coast and the first record in C. corteziensis. The origin of this parasite in the area is unknown, but it may be associated to introductions of Crassostrea virginica from the East coast of United States of America or Gulf of Mexico. PMID- 18423485 TI - Novel bacterial pathogen Acaricomes phytoseiuli causes severe disease symptoms and histopathological changes in the predatory mite Phytoseiulus persimilis (Acari, Phytoseiidae). AB - Adult female Phytoseiulus persimilis Athias-Henriot (Acari, Phytoseiidae) of a laboratory population show a set of characteristic symptoms, designated as non responding (NR) syndrome. Mature predators shrink, cease oviposition and die. They show a lower degree of attraction to herbivore-induced plant volatiles and a greater tendency to leave prey patches carrying ample prey. Moreover, predators may carry excretory crystals in the legs, may cease prey consumption and have a low excretion rate. Here, we satisfy Koch's postulates for a strain of Acaricomes phytoseiuli (DSM 14247) that was isolated from symptomatic female P. persimilis of the NR-population. Adult female P. persimilis were either exposed to a bacterial inoculum suspension (treatment) or to sterile distilled water (control) for a period of 3 days. Control and treated predators were examined for the occurrence of six symptoms characteristic for the NR-syndrome and the presence of A. phytoseiuli after inoculation. The latter was done by re-isolation of A. phytoseiuli from individual predators and predator feces placed on nutrient agar, by PCR-based identification and by histopathological studies of individual predators. The NR-syndrome was clearly induced in those predators that had been exposed to the bacterial inoculum (incubation time=2-5 days, fraction shrunken females=80%), whereas predators exposed to water did not show the NR-syndrome. A. phytoseiuli was never isolated from control predators whereas it could be re isolated from 60% of the treated predators (N=37) and from feces of 41% of treated predators (N=17). Only one day after exposure A. phytoseiuli could not be re-isolated from treated predators and their feces. Light and electron microscope studies of predators exposed to A. phytoseiuli revealed striking bacterial accumulations in the lumen of the alimentary tract together with extreme degeneration of its epithelium. In addition, bacterial foci also occurred in the fat body. These phenomena were not observed in control predators that were exposed to sterile water. The present data prove that A. phytoseiuli can infect the predatory mite P. persimilis and induce the NR-syndrome and characteristic histopathological changes in adult female P. persimilis. This is the first record of a bacterial pathogen in a phytoseiid mite and the first description of pathogenic effects of a bacterial species in the genus Acaricomes. PMID- 18423487 TI - Steric zipper of the amyloid fibrils formed by residues 109-122 of the Syrian hamster prion protein. AB - We report the results of atomic force microscopy, Fourier-transform infrared spectroscopy, solid-state nuclear magnetic resonance, and molecular dynamics (MD) calculations for amyloid fibrils formed by residues 109-122 of the Syrian hamster prion protein (H1). Our data reveal that H1 fibrils contain no more than two beta sheet layers. The peptide strands of H1 fibrils are antiparallel with the A117 residues aligned to form a linear chain in the direction of the fibril axis. The molecular structure of the H1 fibrils, which adopts the motif of steric zipper, is highly uniform in the region of the palindrome sequence AGAAAAGA. The closest distance between the two adjacent beta-sheet layers is found to be about 5 A. The structural features of the molecular model of H1 fibrils obtained by MD simulations are consistent with the experimental results. Overall, our solid state NMR and MD simulation data indicate that a steric zipper, which was first observed in the crystals of fibril-forming peptides, can be formed in H1 fibrils near the region of the palindrome sequence. PMID- 18423488 TI - Improvement of an antibody neutralizing the anthrax toxin by simultaneous mutagenesis of its six hypervariable loops. AB - The enhancement of antibody affinity by mutagenesis targeting only complementarity determining regions has the advantage of respecting the framework regions, which are important for tolerance if clinical use is envisaged. Here, starting from a Fab (antigen-binding fragment; 35PA(83)) capable of neutralizing the lethal toxin of anthrax and having an affinity of 3.4 nM for its antigen, a phage-displayed library of variants where all six complementarity determining regions (73 positions) were targeted for mutagenesis was built. This library contained 5 x 10(8) variants, and each variant carried four mutations on average. The library was first panned with adsorbed antigen and washes of increasing stringency. It was then screened in parallel with either small concentrations of soluble biotinylated antigen or adsorbed antigen and long elution times in the presence of soluble antigen. The stringencies of both selections were pushed as far as possible. Compared with 35PA(83), the best selected clone had an affinity enhanced 19-fold, to 180 pM, and its 50% inhibitory concentration was decreased by 40%. The results of the two selection methods were compared, and the generality of these methods was considered. PMID- 18423486 TI - Insulin signaling pathways and cardiac growth. AB - The development, growth, function and metabolism of the heart are regulated by extracellular growth factors, cytokines and ligands. In this review, the role of insulin and insulin-like growth factors in the regulation of cardiac growth will be discussed. In addition, the role of insulin- and insulin-like growth factor stimulated intracellular signaling proteins in cardiac growth will be reviewed. PMID- 18423489 TI - Porphyrin binding and distortion and substrate specificity in the ferrochelatase reaction: the role of active site residues. AB - The specific insertion of a divalent metal ion into tetrapyrrole macrocycles is catalyzed by a group of enzymes called chelatases. Distortion of the tetrapyrrole has been proposed to be an important component of the mechanism of metallation. We present the structures of two different inhibitor complexes: (1) N methylmesoporphyrin (N-MeMP) with the His183Ala variant of Bacillus subtilis ferrochelatase; (2) the wild-type form of the same enzyme with deuteroporphyrin IX 2,4-disulfonic acid dihydrochloride (dSDP). Analysis of the structures showed that only one N-MeMP isomer out of the eight possible was bound to the protein and it was different from the isomer that was earlier found to bind to the wild type enzyme. A comparison of the distortion of this porphyrin with other porphyrin complexes of ferrochelatase and a catalytic antibody with ferrochelatase activity using normal-coordinate structural decomposition reveals that certain types of distortion are predominant in all these complexes. On the other hand, dSDP, which binds closer to the protein surface compared to N-MeMP, does not undergo any distortion upon binding to the protein, underscoring that the position of the porphyrin within the active site pocket is crucial for generating the distortion required for metal insertion. In addition, in contrast to the wild-type enzyme, Cu(2+)-soaking of the His183Ala variant complex did not show any traces of porphyrin metallation. Collectively, these results provide new insights into the role of the active site residues of ferrochelatase in controlling stereospecificity, distortion and metallation. PMID- 18423490 TI - Effects of risperidone on glucose metabolism in Chinese patients with schizophrenia: a prospective study. AB - BACKGROUND: While most of the second generation antipsychotic agents are associated with abnormal glucose metabolism, previous studies have shown that risperidone has relatively little effect upon blood glucose levels. This study aimed to explore the effect of risperidone on the glucose-regulating mechanism of patients with schizophrenia by using the oral glucose tolerance test (OGTT), measuring insulin and C-peptide levels. METHODS: Thirty inpatients with schizophrenia taking risperidone were studied. All the patients were given a simplified OGTT at baseline and six weeks after treatment. Plasma glucose, insulin, and C-peptide concentrations were measured at fasting, then 1 and 2h after OGTT respectively. Other data, including demographic characteristics and plasma drug concentrations, were also recorded. RESULTS: (1) There was no significant increase in the proportion of patients demonstrating abnormal plasma glucose levels compared with baseline (p=1.000, McNemar test); (2) risperidone was associated with elevated insulin concentrations (p=0.013), C-peptide levels (p=0.020), insulin/glucose ratio (p=0.020) and BMI (p<0.01); (3) no sex differences in glucose-related measures were observed. CONCLUSION: Risperidone treatment may be associated with alterations in glucose-regulating mechanisms in patients with schizophrenia. PMID- 18423491 TI - Natural selection of cooperation and degree hierarchy in heterogeneous populations. AB - One of the current theoretical challenges to the explanatory powers of Evolutionary Theory is the understanding of the observed evolutionary survival of cooperative behavior when selfish actions provide higher fitness (reproductive success). In unstructured populations natural selection drives cooperation to extinction. However, when individuals are allowed to interact only with their neighbors, specified by a graph of social contacts, cooperation-promoting mechanisms (known as lattice reciprocity) offer to cooperation the opportunity of evolutionary survival. Recent numerical works on the evolution of Prisoner's Dilemma in complex network settings have revealed that graph heterogeneity dramatically enhances the lattice reciprocity. Here we show that in highly heterogeneous populations, under the graph analog of replicator dynamics, the fixation of a strategy in the whole population is in general an impossible event, for there is an asymptotic partition of the population in three subsets, two in which fixation of cooperation or defection has been reached and a third one which experiences cycles of invasion by the competing strategies. We show how the dynamical partition correlates with connectivity classes and characterize the temporal fluctuations of the fluctuating set, unveiling the mechanisms stabilizing cooperation in macroscopic scale-free structures. PMID- 18423492 TI - Predicting protein structural class by SVM with class-wise optimized features and decision probabilities. AB - Determination of protein structural class solely from sequence information is a challenging task. Several attempts to solve this problem using various methods can be found in literature. We present support vector machine (SVM) approach where probability-based decision is used along with class-wise optimized feature sets. This approach has two distinguishing characteristics from earlier attempts: (1) it uses class-wise optimized features and (2) decisions of different SVM classifiers are coupled with probability estimates to make the final prediction. The algorithm was tested on three datasets, containing 498 domains, 1092 domains and 5261 domains. Ten-fold external cross-validation was performed to assess the performance of the algorithm. Significantly high accuracy of 92.89% was obtained for the 498-dataset. We achieved 54.67% accuracy for the dataset with 1092 domains, which is better than the previously reported best accuracy of 53.8%. We obtained 59.43% prediction accuracy for the larger and less redundant 5261 dataset. We also investigated the advantage of using class-wise features over union of these features (conventional approach) in one-vs.-all SVM framework. Our results clearly show the advantage of using class-wise optimized features. Brief analysis of the selected class-wise features indicates their biological significance. PMID- 18423493 TI - An optimization framework of biological dynamical systems. AB - Different biological dynamics are often described by different mathematical equations. On the other hand, some mathematical models describe many biological dynamics universally. Here, we focus on three biological dynamics: the Lotka Volterra equation, the Hopfield neural networks, and the replicator equation. We describe these three dynamical models using a single optimization framework, which is constructed with employing the Riemannian geometry. Then, we show that the optimization structures of these dynamics are identical, and the differences among the three dynamics are only in the constraints of the optimization. From this perspective, we discuss the unified view for biological dynamics. We also discuss the plausible categorizations, the fundamental nature, and the efficient modeling of the biological dynamics, which arise from the optimization perspective of the dynamical systems. PMID- 18423494 TI - Predicting protein structural class based on multi-features fusion. AB - Structural class characterizes the overall folding type of a protein or its domain and the prediction of protein structural class has become both an important and a challenging topic in protein science. Moreover, the prediction itself can stimulate the development of novel predictors that may be straightforwardly applied to many other relational areas. In this paper, 10 frequently used sequence-derived structural and physicochemical features, which can be easily computed by the PROFEAT (Protein Features) web server, were taken as inputs of support vector machines to develop statistical learning models for predicting the protein structural class. More importantly, a strategy of merging different features, called best-first search, was developed. It was shown through the rigorous jackknife cross-validation test that the success rates by our method were significantly improved. We anticipate that the present method may also have important impacts on boosting the predictive accuracies for a series of other protein attributes, such as subcellular localization, membrane types, enzyme family and subfamily classes, among many others. PMID- 18423495 TI - Preliminary health risk assessment for polybrominated diphenyl ethers and polybrominated dibenzo-p-dioxins/furans in seafood from Guangzhou and Zhoushan, China. AB - Dietary intake is one of the important routes of human exposure to brominated flame retardants (BFRs) such as polybrominated diphenyl ethers (PBDEs). The use of PBDEs may also result in exposure to polybrominated dibenzo-p-dioxins and dibenzofurans (PBDDs/DFs), as these compounds are impurities in technical mixtures of BFRs and can also be formed unintentionally by the same processes that generate chlorinated dioxins. This study determined the concentrations of polybrominated compounds in common seafood in Guangzhou and Zhoushan, and assessed the health risks of these chemicals via consumption of contaminated seafood. Seafood samples (fish, bivalves, shrimp, crab, and cephalopods) purchased from local markets in 2003 and 2004 were analyzed for PBDEs and PBDDs/DFs. The highest concentration of total PBDEs (46.3 ng g(-1) lipid wt.) was detected in fish from Guangzhou, in which BDEs 47 and 209 were the two predominant congeners. The total daily intakes of PBDEs, PBDDs, and PBDFs were, 946, 6.39, and 6.54 pg kg(-1) body weight (bw) in Guangzhou, and 489, 4.99, and 7.65 pg kg(-1) bw in Zhoushan, respectively. The hazard ratios for PBDDs and PBDFs were both greater than unity, indicating that these compounds may pose some health risks to the local population. PMID- 18423496 TI - Cloning and developmental expression of kinesin superfamily7 (kif7) in the brackish medaka (Oryzias melastigma), a close relative of the Japanese medaka (Oryzias latipes). AB - kif7 is a member of the kinesin superfamily members which are molecular motor proteins that move along microtubules in a highly regulated manner through ATP hydrolysis. In this paper, we report on the cloning of the Oryziasmelastigmakif7 (omkif7) using primers designed according to the Japanese medaka (Oryziaslatipes) database. The cloned omkif7 has an open reading frame of 3762bp and is deduced to encode a polypeptide of 1254 amino acids that possesses the putative ATP-binding and microtubule-binding motifs in its motor domain at the N-terminal region. We characterized the cloned omkif7 by comparison with the zebrafish kif7. Both omkif7 and zebrafish kif7 are shown to be expressed in all embryonic stages and adult tissues examined with higher expression level in the testis and ovary. Whole-mount in situ hybridization revealed that the expression of omkif7 is ubiquitous during the early stages of embryonic development, but became more restrictive and localized to the brain, fin bud and eye at later development. This study suggested that the brackish O.melastigma can serve as a good seawater model organism for developmental studies by utilizing the resources developed from its close relative of the Japanese medaka. PMID- 18423497 TI - Use of microcirculatory parameters to evaluate clinical treatments of chronic venous disorder (CVD). AB - OBJECTIVES: To evaluate changes on cutaneous microangiopathy in chronic venous disorder (CVD) after use of Cirkan [venotonic drug containing Ruscus aculeatus (plant extract), hesperidine methylchalcone (flavonoid) and vitamin C], elastic compression stockings (ECS) or no treatment for four weeks. PATIENTS AND METHODS: Fifty-five female patients (85 legs), 25 to 57 years, with at least one limb classified as C2,s or C2,3,s (CEAP classification), were allocated consecutively, according to entrance order, in these three groups. Ten healthy women age-matched were also investigated. Using orthogonal polarization spectral technique (noninvasive method), measurements of functional capillary density (FCD, number of capillaries with flowing red blood cells/mm(2)), capillary morphology (CM, % of abnormal capillaries/mm(2)) and diameters (mum) of dermal papilla (DDP), capillary bulk (DCB) and capillary limb (CD) were obtained on the medial perimalleolar region and later analyzed using CapImage software. RESULTS AND CONCLUSIONS: CVD patients showed significant changes on CD and CM compared to healthy subjects in agreement with our previous findings (J Vasc Surg 43:1037 1044, 2006). On Cirkan-treated patients, after 4 weeks, CD decreased on both limbs and CM improved on the left one, suggesting an amelioration of the chronic venous hypertension. No significant changes could be detected on other patient groups. These results confirm the existence of microcirculatory dysfunction in early stages of CVD, probably due to post-capillary hypertension, and further support the venotonic action of Cirkan. PMID- 18423499 TI - No somatic mutations detected in the Mad2 gene in 658 human tumors. PMID- 18423498 TI - Assessment of DNA double-strand breaks and gammaH2AX induced by the topoisomerase II poisons etoposide and mitoxantrone. AB - Double-strand breaks (DSBs) are highly deleterious DNA lesions as they lead to chromosome aberrations and/or apoptosis. The formation of nuclear DSBs triggers phosphorylation of histone H2AX on Ser-139 (defined as gammaH2AX), which participates in the repair of such DNA damage. Our aim was to compare the induction of gammaH2AX in relation to DSBs induced by topoisomerase II (TOPO II) poisons, etoposide (ETOP) and mitoxantrone (MXT), in V79 cells. DSBs were measured by the neutral comet assay, while gammaH2AX was quantified using immunocytochemistry and flow cytometry. Stabilized cleavage complexes (SCCs), lesions thought to be responsible for TOPO II poison-induced genotoxicity, were measured using a complex of enzyme-DNA assay. In the case of ETOP, a no observed adverse effect level (NOAEL) and lowest observed effect level (LOEL) for genotoxicity was determined; gammaH2AX levels paralleled DSBs at all concentrations but significant DNA damage was not detected below 0.5 microg/ml. Furthermore, DNA damage was dependent on the formation of SCCs. In contrast, at low MXT concentrations (0.0001-0.001 microg/ml), induction of gammaH2AX was not accompanied by increases in DSBs. Rather, DSBs were only significantly increased when SCCs were detected. These findings suggest MXT-induced genotoxicity occurred via at least two mechanisms, possibly related to DNA intercalation and/or redox cycling as well as TOPO II inhibition. Our findings also indicate that gammaH2AX can be induced by DNA lesions other than DSBs. In conclusion, gammaH2AX, when measured using immunocytochemical and flow cytometric methods, is a sensitive indicator of DNA damage and may be a useful tool in genetic toxicology screens. ETOP data are consistent with the threshold concept for TOPO II poison-induced genotoxicity and this should be considered in the safety assessment of chemicals displaying an affinity for TOPO II and genotoxic/clastogenic effects. PMID- 18423500 TI - [Malformation of cortical development: which strategy is best?]. AB - Malformations of cortical development (MCD) correspond to a broad spectrum of cerebral lesions resulting from cortical development abnormalities during embryogenesis. They are frequently associated with drug-resistant epilepsy as well as more or less severe neurological and cognitive deficits. Diagnosis of MCD has greatly improved with the progress in contemporary imaging techniques, and patients with cryptogenic epilepsy are increasingly recognized as having MCD. Current classifications based on the combination of clinical, imaging, genetic and pathological data allow analysis of homogeneous patient series and optimal therapeutic strategies. Successful surgical treatment can be proposed to patients with focal lesions such as focal cortical dysplasia or tumors associated with cortical dysplasia (dysembryoplastic neuroepithelial tumors and gangliogliomas). Favorable outcome can also be obtained in some diffuse cases such as tuberous sclerosis, periventricular heterotopia and polymicrogyria. Invasive monitoring, especially stereoelectroencephalography (SEEG), has proved to be useful in determining the organization of the epileptogenic zone in each MCD type and planning cortical resections. Moreover, establishing correlations between neurophysiological data, imaging and pathological findings has allowed surgery without previous invasive procedures in the majority of focal MCD types; however, intracranial recordings remain necessary in case of more diffuse MCD. PMID- 18423501 TI - [Surgical procedures for epilepsy. Introduction]. PMID- 18423502 TI - [Surgical resection of focal cortical dysplasias in the central region]. AB - BACKGROUND AND PURPOSE: Taylor-type focal cortical dysplasias (TTFCD) represent a particular pathological entity responsible for severe drug-resistant epilepsy of extratemporal location. Epilepsy can be surgically cured if complete removal of the lesion can be performed. However, identification on imaging may be difficult and negative standard MRIs are not rare. The frequent location of TTFCD in the central region restrains the possibilities of complete resection. We report a series of patients operated on for intractable epilepsy associated with TTFCD in the central area. PATIENTS AND METHODS: Between 2000 and 2006, of 34 consecutive patients with TTFCD, 17 had a lesion located in the central area. MRI was considered normal in eight, although in five a subtle gyral abnormality was disclosed on further analysis. A (18)FDG PET scan performed in 16 cases demonstrated focal hypometabolism in 15 that correlated with abnormalities on MRI when visible. SEEG performed in 13 cases revealed typical abnormalities for TTFCD in 10 cases. At resection, cortical and subcortical stimulations of the dysplastic cortex did not elicit a motor response. RESULTS: Postoperative motor or sensory deficit was observed in 13 patients--severe in four--which subsequently resolved completely in seven. Six patients had a minor permanent, motor or sensory deficit. Four patients were reoperated for seizure recurrence and residual dysplastic tissue was found at reoperation in three cases. Average postoperative follow-up was 3.7 years. Sixteen patients (94%) were in Engel Class I (65% in Class IA). CONCLUSION: This study suggests that surgical resection of central region TTFCD may be associated with favorable seizure outcome and no or minor functional permanent disability. In cases of seizure relapse, reoperation can be performed without further permanent deficit and lead to seizure-free outcome. Future techniques for intraoperative detection of these lesions could optimize their complete resection in functional areas. PMID- 18423503 TI - Evaluation of dietary and management risk factors for enterolithiasis among horses in California. AB - Enteroliths are intestinal calculi that result in intestinal obstruction and colic in horses. Equine enterolithiasis occurs worldwide, but the disease is particularly prevalent in some geographic locations, including California. The objectives of this study were to evaluate dietary and environmental risk factors for the disease. This was accomplished through a case-control study by comparing horses with colic from enterolithiasis presenting to the University of California, Davis VMTH, to horses with colic of other causes. Data were collected on 61 horses with enterolithiasis and 75 controls via evaluation of patient records and questionnaires completed by owners at the time of admission. Following multiple logistic regression analysis, the following factors were found to be significantly associated with enterolithiasis: feeding > or = 50% of the diet as alfalfa; feeding <50% of the diet as oat hay; feeding <50% of the diet as grass hay; and lack of daily access to pasture grazing. PMID- 18423504 TI - The role of hygienic bathing after defecation in the transmission of Schistosoma mansoni. AB - Transmission of Schistosoma mansoni depends on fecal eggs reaching water, but the way this happens is poorly understood. We studied the role of hygienic bathing after defecation in the contamination of water with S. mansoni eggs. Individuals in an endemic community in Northern Senegal (n=991) were examined for S. mansoni infection and a random sample (22%) was interviewed about stool disposal practices and hygienic behavior. We assessed the presence and viability of S. mansoni eggs adhering to the peri-anal region of 13 infected volunteers, by counting the miracidia in the water they had used for hygienic washing; for 10 of them (77%) miracidia were demonstrated. From the population infection distribution, average number of defecations per day, proportion of individuals bathing after defecation, and association between miracidial counts and infection intensity, we calculated a daily population miracidial output of approximately 30,000 through hygienic bathing. For comparison, one complete stool reaching the water was calculated to yield approximately 2500 miracidia. Thus, 12 individuals in this population should defecate into the water every day to produce the same number of miracidia as through hygienic bathing. Our results suggest a major role of hygienic bathing after defecation in the transmission of S. mansoni. PMID- 18423505 TI - Synthesis of exemestane labelled with (13)C. AB - The synthesis of exemestane Aromasin, an irreversible steroidal aromatase inhibitor, specifically labelled with (13)C is reported. The preparation of [(13)C(3)]exemestane was achieved according to an eight-step procedure starting from the commercially available testosterone. PMID- 18423506 TI - Altered binding of thioflavin t to the peripheral anionic site of acetylcholinesterase after phosphorylation of the active site by chlorpyrifos oxon or dichlorvos. AB - The peripheral anionic site of acetylcholinesterase, when occupied by a ligand, is known to modulate reaction rates at the active site of this important enzyme. The current report utilized the peripheral anionic site specific fluorogenic probe thioflavin t to determine if the organophosphates chlorpyrifos oxon and dichlorvos bind to the peripheral anionic site of human recombinant acetylcholinesterase, since certain organophosphates display concentration dependent kinetics when inhibiting this enzyme. Incubation of 3 nM acetylcholinesterase active sites with 50 nM or 2000 nM inhibitor altered both the B(max) and K(d) for thioflavin t binding to the peripheral anionic site. However, these changes resulted from phosphorylation of Ser203 since increasing either inhibitor from 50 nM to 2000 nM did not alter further thioflavin t binding kinetics. Moreover, the organophosphate-induced decrease in B(max) did not represent an actual reduction in binding sites, but instead likely resulted from conformational interactions between the acylation and peripheral anionic sites that led to a decrease in the rigidity of bound thioflavin t. A drop in fluorescence quantum yield, leading to an apparent decrease in B(max), would accompany the decreased rigidity of bound thioflavin t molecules. The organophosphate-induced alterations in K(d) represented changes in binding affinity of thioflavin t, with diethylphosphorylation of Ser203 increasing K(d), and dimethylphosphorylation of Ser203 decreasing K(d). These results indicate that chlorpyrifos oxon and dichlorvos do not bind directly to the peripheral anionic site of acetylcholinesterase, but can affect binding to that site through phosphorylation of Ser203. PMID- 18423507 TI - Antioxidant status of erythrocytes and their response to oxidative challenge in humans with argemone oil poisoning. AB - Oxidative damage of biomolecules and antioxidant status in erythrocytes of humans from an outbreak of argemone oil (AO) poisoning in Kannauj (India) and AO intoxicated experimental animals was investigated. Erythrocytes of the dropsy patients and AO treated rats were found to be more susceptible to 2,2'-azobis (2 amidinopropane) dihydrochloride (AAPH) induced peroxidative stress. Significant decrease in RBC glutathione (GSH) levels (46, 63%) with concomitant enhancement in oxidized glutathione (172, 154%) levels was noticed in patients and AO intoxicated animals. Further, depletion of glutathione reductase (GR), glucose-6 phosphate dehydrogenase (G-6-PDH) and glutathione-S-transferase (GST) (42-52%) was observed in dropsy patients. Oxidation of erythrocyte membrane lipids and proteins was increased (120-144%) in patients and AO treated animals (112-137%) along with 8-OHdG levels in whole blood (180%) of dropsy patients. A significant reduction in alpha-tocopherol content (68%) was noticed in erythrocytes of dropsy patients and hepatic, plasma and RBCs of AO treated rats (59-70%) thereby indicating the diminished antioxidant potential to scavenge free radicals or the limited transport of alpha-tocopherol from liver to RBCs leading to enhanced oxidation of lipids and proteins in erythrocytes. These studies implicate an important role of erythrocyte degradation in production of anemia and breathlessness in epidemic dropsy. PMID- 18423508 TI - A zebrafish (Danio rerio) model of infectious spleen and kidney necrosis virus (ISKNV) infection. AB - Zebrafish is a model animal for studies of genetics, development, toxicology, oncology, and immunology. In this study, infectious spleen and kidney necrosis virus (ISKNV) was used to establish an infection in zebrafish, and the experimental conditions were established and characterized. Mortality of adult zebrafish infected with ISKNV by intraperitoneal (i.p.) injection exceeded 60%. ISKNV can be passed stably in zebrafish for over ten passages. The ailing zebrafish displayed petechial hemorrhaging and scale protrusion. Histological analysis of moribund fish revealed necrosis of tissue and enlarged cells in kidney and spleen. The real-time RT-PCR analysis of mRNA level confirmed that ISKNV was replicated in zebrafish. Immunohistochemistry and immunofluorescence analyses further confirmed the presence of ISKNV-infected cells in almost all organs of the infected fish. Electron microscope analyses showed that the ISKNV particle was present in the infected tissues. The establishment of zebrafish infection model of ISKNV can offer a valuable tool for studying the interactions between ISKNV and its host. PMID- 18423510 TI - Changes in the intraisolate genetic structure of Beet necrotic yellow vein virus populations associated with plant resistance breakdown. AB - The causal agent of rhizomania disease, Beet necrotic yellow vein virus (BNYVV), typically produces asymptomatic root-limited infections in sugar beets (Beta vulgaris) carrying the Rz1-allele. Unfortunately, this dominant resistance has been recently overcome. Multiple cDNA clones of the viral pathogenic determinant p25, derived from populations infecting susceptible or resistant plants, were sequenced to identify host effects on the viral population structure. Populations isolated from compatible plant-virus interactions (susceptible plant-wild type virus and resistant plant-resistant breaking variants) were large and relatively homogeneous, whereas those from the incompatible interaction (resistant plant avirulent type virus) were small and highly heterogeneous. All populations from susceptible plants had the same dominant haplotype, whereas those from resistant cultivars had a different haplotype surrounded by a spectrum of mutants. Selection and diversification analyses suggest an evolutionary trajectory of BNYVV with positive selection for changes required to overcome resistance, followed by elimination of hitchhiking mutations through purifying selection. PMID- 18423509 TI - Nuclear trafficking of the human cytomegalovirus pp71 (ppUL82) tegument protein. AB - The human cytomegalovirus tegument protein pp71 localizes to the nucleus immediately upon infection, and functions to initiate viral gene expression. Analysis of a series of random insertion mutations revealed that sequences within the mid region (MR) of pp71 are important for localization to the nucleus. Fusion of MR sequences with eGFP revealed that amino acids 94 to 300 were sufficient to target proteins to the nucleus. Random substitution mutagenesis within this domain resulted in two double substitution mutants, pp71P203T/T223M and pp71T228M/L275Q, with a predominantly cytoplasmic localization. Disruption of nuclear targeting resulted in relocalization of the fusion proteins to a distinct perinuclear region. Using tandem mass spectrometry, we determined that threonine 223 can be phosphorylated. Mutation of this residue to a phosphomimetic amino acid resulted in abrogation of nuclear targeting. These results strongly suggest that the intracellular trafficking of pp71 is regulated by phosphorylation. PMID- 18423512 TI - A novel method for on-line evaluation of floc size in coagulation process. AB - Chemical coagulation is a simple and widely used water treatment process. A jar test based on the residual turbidity in the treated water was used to evaluate the optimal conditions for floc formation. However, the final residual turbidity does not show up variation of turbidity and floc formation during the flocculation process. Hence, a nephelometric turbidimeter method based on on-line monitoring was devised to determine the floc size variance during flocculation. A nephelometric turbidimeter coupled with a data acquisition unit was used to measure turbidity every second at 3 cm below the water surface during the coagulation process. Laboratory results indicated that this new instrument was capable of recording floc agglomeration during slow mixing very accurately. The standard deviation (SD) of the measured turbidity was proportional to the square root of the floc size; a greater SD indicated larger floc sizes. Hence, in addition to monitoring turbidity, the nephelometric turbidity meter is also a valuable tool to study the floc agglomeration process and variations in the resulting floc size. This method is simple and effective; it contributes significantly to the selection of coagulant and optimal flocculation conditions to improve water treatment. PMID- 18423511 TI - Protanomaly without darkened red is deuteranopia with rods. AB - The Rayleigh match, a color match between a mixture of 545+670 nm lights and 589 nm light in modern instruments, is the definitive measurement for the diagnosis of inherited red-green color defects. All trichromats, whether normal or anomalous, have a limited range of 545+670 nm mixtures they perceive to match 589 nm: a typical color-normal match range is about 50-55% of 670 nm in the mixture (deutan mode), while deuteranomals have a range that includes mixtures with less 670 nm than normal and protanomals a range that includes mixtures with more 670 nm than normal. Further, the matching luminance of the 589 nm light for deuteranomals is the same as for normals but for protanomals is below normal. An example of an unexpected Rayleigh match, therefore, is a match range above normal (typical of protanomaly) and a normal luminance setting for 589 nm (typical of deuteranomaly), a match called protanomaly "when the red end of the spectrum is not darkened" [Pickford, R.W. (1950). Three pedigrees for color blindness. Nature, 165, 182.]. In this case, Rayleigh matching does not yield a clear diagnosis. Aside from Pickford, we are aware of only one other report of a similar observer [Pokorny, J., & Smith, V. C. (1981). A variant of red-green color defect. Vision Research, 21, 311-317]; this study predated modern genetic techniques that can reveal the cone photopigment(s) in the red-green range. We recently had the opportunity to conduct genetic and psychophysical tests on such an observer. Genetic results predict he is a deuteranope. His Rayleigh match is consistent with L cones and a contribution from rods. Further, with a rod suppressing background, his Rayleigh match is characteristic of a single L-cone photopigment (deuteranopia). PMID- 18423513 TI - New biosorbent materials for heavy metal removal: product development guided by active site characterization. AB - Olive pomace wastes were used as precursors for the development of new biosorbents for heavy metals. Thermal and/or chemical treatments were addressed in terms of their effects on functional group properties and copper removal. Product development was guided by experimental characterization (potentiometric titrations, IR spectra, copper biosorption, total surface area) and theoretical modeling of acid-base properties. Olive pomace was characterized by negative charge due to dissociation of two weakly acidic sites (carboxylic and phenolic). After thermal treatment, a char-like material was obtained, characterized by basic sites with positive charge in the acidic pH range. Copper biosorption properties of this material were improved with respect to olive pomace due to the binding ability of pyrone-like sites. Both native and carbonized olive pomace samples were chemically treated by HNO(3). Experimental tests were performed according to factorial designs and analysis of variance was used to evaluate significant effects on copper removal of both oxidant concentration and temperature. Chemical treatment by HNO(3) of carbonized material developed negatively charged groups in the acidic pH range (carboxylic groups), but presented a decreased copper removal. Chemical treatment by HNO(3) of olive pomace without carbonization gave biosorbents with acid-base properties similar to those of native olive pomace, but with a significant increase of copper biosorption. Specific carboxylic-copper interactions indicated by IR spectra and increased surface area due to chemical treatment were simultaneously responsible for such improvement. PMID- 18423514 TI - Model description and kinetic parameter analysis of MTBE biodegradation in a packed bed reactor. AB - A dynamic modeling approach was used to estimate in-situ model parameters, which describe the degradation of methyl tert-butyl ether (MTBE) in a laboratory packed bed reactor. The measured dynamic response of MTBE pulses injected at the reactor's inlet was analyzed by least squares and parameter response surface methodologies. Response surfaces were found to be statistically significant and thus suitable for estimating the global minimum as well as the 95% parameter uncertainty regions. The linear parameter uncertainty estimates for the half saturation constant (K(S)) and the maximum growth rate (micro(max)) were: 05) and beta-(1-->6) galactofuranosyl residues. Galactan biosynthesis is essential for mycobacterial viability and thus inhibitors of the enzymes involved in its assembly are potential drugs for the treatment of mycobacterial diseases, including tuberculosis. Only two galactofuranosyltransferases, GlfT1 and GlfT2, are responsible for the biosynthesis of the entire galactan domain of the mAG and we report here the first high-throughput assay for GlfT2. Successful implementation of the assay required the synthesis of multi-milligram amounts of the donor for the enzyme, UDP-Galf, 1, which was achieved using a chemoenzymatic approach. We also describe an improved expression system for GlfT2, which provides a larger amount of active protein for the assay. Kinetic analysis of 1 and a known trisaccharide acceptor for the enzyme, 2, have been carried out and the apparent K(m) and k(cat) values obtained for the latter are in agreement with those obtained using a previously reported radiochemical assay. The assay has been implemented in 384-well microtiter plates, which will facilitate the screening of large numbers of potential GlfT2 inhibitors, with possible utility as novel anti TB drugs. PMID- 18423587 TI - Assessment of water quality of polluted lake using multivariate statistical techniques: a case study. AB - Multivariate statistical techniques, cluster analysis (CA) and principal component analysis (PCA) were applied to the data on water quality of Manchar Lake (Pakistan), generated during 2005-06, with monitoring at five different sites for 36 parameters. This study evaluated and interpreted complex water quality data sets and apportioned of pollution sources to get better information about water quality and to design a monitoring network. The chemical correlations were observed by PCA, which were used to classify the samples by CA, based on the PCA scores. Three significant sampling locations--(sites 1 and 2), (site 4) and (sites 3 and 5)--were detected on the basis of similarity of their water quality. The results revealed that the major causes of water quality deterioration were related to inflow of effluent from industrial, domestic, agricultural and saline seeps into the lake at site 1 and also resulting from people living in boats and fishing at sites 2 and 3. PMID- 18423588 TI - Acute effects of 1-octyl-3-methylimidazolium bromide ionic liquid on the antioxidant enzyme system of mouse liver. AB - The effects of acute exposure of intraperitoneal injection of aqueous 1-octyl-3 methylimidazolium bromide on the antioxidant enzymes and lipid peroxidation of the treated mouse liver were investigated in the present paper. Thirty mice were treated with 17.9 (0.5 median lethal dose, LD(50)) and 35.7 mg/kg (1 LD(50)) of the ionic liquid or saline solution by intraperitoneal injection for 10h. Significant increase in hepatosomatic index and decrease in liver protein content were found in 1 LD(50) group. No statistically significant change was observed in the activity of superoxide dismutase in both treated groups compared to the control group. In the case of catalase activity, the decrease was recorded only at dose of 1 LD(50). As for the activity of glutathione peroxidase, increase in 0.5 LD(50) group while significant decrease at dose of 1 LD(50) was noted. There was an increase in the activity of glutathione-S-transferase in these two treated groups in comparison to the control. However, no effect on the level of lipid peroxidation was found in the treated groups. These results showed that the acute exposure of aqueous ionic liquid could cause damage to mouse, leading to antioxidant responses in livers of the treated mice. PMID- 18423589 TI - Mycobacterium bovis Bacillus Calmette-Guerin (BCG) stimulates IL-10 production via the PI3K/Akt and p38 MAPK pathways in human lung epithelial cells. AB - Infection of human cells with mycobacteria has been shown to result in the production of anti-inflammatory cytokines. However, the signaling pathways that regulate the Mycobacterium bovis BCG-induced interleukin (IL)-10 production are currently unknown. In the present study, we investigated the involvement of phosphatidylinoditol 3-kinase (PI3K)/Akt and the p38 MAPK signaling pathways in the secretion of IL-10 in human lung epithelial cells (A549) after infection with M. bovis BCG. Treatment of A549 cells with LY 294002 (2-(4-morpholinyl)-8-phenyl 4H-1-benzopyran-4-one) and wortmannin, two PI3K inhibitors, inhibited M. bovis BCG-induced IL-10 production. Stimulation of cells with M. bovis BCG caused an increase in Akt phosphorylation in a time-dependent manner, which was inhibited by wortmannin. In addition, treatment of A549 cells with an Akt inhibitor significantly blocked M. bovis BCG-induced IL-10 production. Moreover, the p38 inhibitor SB203580 significantly decreased IL-10 production in a dose-dependent manner, whereas M. bovis BCG-induced IL-10 secretion was completely unaffected by the MEK inhibitor PD98059. Finally, the inhibition of PI3K did not significantly affect p38 MAPK activation in M. bovis BCG-infected cells, indicating that PI3K activity is not required for the M. bovis BCG-induced phosphorylation of p38 MAPK. Collectively, these data suggest that the PI3K/Akt and p38 MAPK signaling pathways play an important role in the regulation of M. bovis BCG-induced IL-10 secretion in A549 cells. PMID- 18423590 TI - Experiential and genetic contributions to depressive- and anxiety-like disorders: clinical and experimental studies. AB - Stressful events have been implicated in the precipitation of depression and anxiety. These disorders may evolve owing to one or more of an array of neuronal changes that occur in several brain regions. It seems likely that these stressor provoked neurochemical alterations are moderated by genetic determinants, as well as by a constellation of experiential and environmental factors. Indeed, animal studies have shown that vulnerability to depressive-like behaviors involve mechanisms similar to those associated with human depression (e.g., altered serotonin, corticotropin releasing hormone and their receptors, growth factors), and that the effects of stressors are influenced by previous stressor experiences, particularly those encountered early in life. These stressor effects might reflect sensitization of neuronal functioning, phenotypic changes of processes that lead to neurochemical release or receptor sensitivity, or epigenetic processes that modify expression of specific genes associated with stressor reactivity. It is suggested that depression is a life-long disorder, which even after effective treatment, has a high rate of re-occurrence owing to sensitized processes or epigenetic factors that promote persistent alterations of gene expression. PMID- 18423591 TI - Behavioural and neurochemical effects of post-weaning social isolation in rodents relevance to developmental neuropsychiatric disorders. AB - Exposing mammals to early-life adverse events, including maternal separation or social isolation, profoundly affects brain development and adult behaviour and may contribute to the occurrence of psychiatric disorders, such as depression and schizophrenia in genetically predisposed humans. The molecular mechanisms underlying these environmentally induced developmental adaptations are unclear and best evaluated in animal paradigms with translational salience. Rearing rat pups from weaning in isolation, to prevent social contact with conspecifics, produces reproducible, long-term changes including; neophobia, impaired sensorimotor gating, aggression, cognitive rigidity, reduced prefrontal cortical volume and decreased cortical and hippocampal synaptic plasticity. These alterations are associated with hyperfunction of mesolimbic dopaminergic systems, enhanced presynaptic dopamine (DA) and serotonergic (5-HT) function in the nucleus accumbens (NAcc), hypofunction of mesocortical DA and attenuated 5-HT function in the prefrontal cortex and hippocampus. These behavioural, morphological and neurochemical abnormalities, as reviewed herein, strongly resemble core features of schizophrenia. Therefore unravelling the mechanisms that trigger these sequelae will improve our knowledge of the aetiology of neurodevelopmental psychiatric disorders, enable identification of longitudinal biomarkers of dysfunction and permit predictive screening for novel compounds with potential antipsychotic efficacy. PMID- 18423592 TI - The long-term behavioural consequences of prenatal stress. AB - Maternal distress during pregnancy increases plasma levels of cortisol and corticotrophin releasing hormone in the mother and foetus. These may contribute to insulin resistance and behaviour disorders in their offspring that include attention and learning deficits, generalized anxiety and depression. The changes in behaviour, with or independent of alterations in the function of the hypothalamic pituitary adrenal (HPA) axis, can be induced by prenatal stress in laboratory rodents and non-human primates. The appearance of such changes depends on the timing of the maternal stress, its intensity and duration, gender of the offspring and is associated with structural changes in the hippocampus, frontal cortex, amygdala and nucleus accumbens. The dysregulation of the HPA axis and behaviour changes can be prevented by maternal adrenalectomy. However, only the increased anxiety and alterations in HPA axis are re-instated by maternal injection of corticosterone. CONCLUSION: Excess circulating maternal stress hormones alter the programming of foetal neurons, and together with genetic factors, the postnatal environment and quality of maternal attention, determine the behaviour of the offspring. PMID- 18423593 TI - Expression and functional analysis of Gm114, a putative mammalian ortholog of Drosophila bam. AB - In the testis, the continuous production of sperm is maintained by a small population of stem cells called germ line stem cells (GSCs) in Drosophila, or spermatogonial stem cells (SSCs) in mammals. This stem cell population can self renew or produce daughter cells that differentiate into mature sperm. In Drosophila, BMP signals inhibit GSC differentiation by blocking transcription of the gene bag of marbles (bam). Once bam is activated, germ cells initiate differentiation. We identified a novel gene in mouse, Gm114, that shows homology to Drosophila bam. In male germ cells, expression of Gm114 begins at 12.5-13.5 days post coitum (dpc), the stage in mice when germ cells cease proliferation and begin differentiation into prospermatogonia. In the adult testis, Gm114 is highly expressed in differentiated spermatocytes and spermatids but not in undifferentiated spermatogonia, strongly suggesting that, similar to Bam, GM114 plays an important role in mammalian germ line stem cell self-renewal and differentiation. Interestingly, deletion of the majority of the GM114 protein does not affect mouse viability or fertility. This suggests that either there is a function for the remaining N-terminal of GM114, or that there are alternative mechanisms in the mammalian system that control germ cell differentiation. PMID- 18423595 TI - Occurrence, distribution and possible sources of organochlorine pesticide residues in tropical coastal environment of India: an overview. AB - Organochlorine pesticides (OCPs) are an important potential component of chemical pollutants used extensively for agriculture and sanitation purposes in India as these are comparatively cheap and effective. These persistent organic compounds such as HCH isomers, DDT and its metabolites are the predominant chemical contaminants found along the Indian coast and thus constitute both alluring and grave areas of scientific research. Our objective in the paper is to provide a comprehensive account of the distribution of organochlorine pesticides in biotic and abiotic compartments of the Indian coastal environment, make some comments on their environmental sources, their movement through the food chain and possible ecotoxicological risk of health in biota including humans. The prevalent HCH, DDT and HCB concentrations differ markedly in eastern and western coast of India reflecting differing agricultural and other usage and their ultimate input into the coastal environment by several rivers and the bioturbation activities of macrozoobenthos (bivalve mollusks, polychaetous annelids, etc.). In several cases, the DDT levels exceeded the effects range-low (ER-L) and could thus cause acute biological impairments, in comparison with the sediment quality guidelines. Contributions of DDT metabolites (DDT, DDD and DDE residues) vary in different Indian coastal regions predominated by pp'-DDT and pp'-DDD. Measured concentrations of HCHs were lower than DDTs that might be due to higher water solubility, vapor pressure and biodegradability of the latter. HCH and DDT residues in fish in India were lower than those in the temperate countries indicating a lower accumulation in tropical fish, which might be related to rapid volatilization of this insecticide in the tropical environment. The concentrations of other chlorinated pesticides (aldrin, dieldrin, eldrin, methoxychlor, endosulfan sulphate) were lower and not generally of great concern. PMID- 18423596 TI - Assessing inhalatory and dermal exposures and their resultant health-risks for workers exposed to polycyclic aromatic hydrocarbons (PAHs) contained in oil mists in a fastener manufacturing industry. AB - This study first assessed workers' inhalatory and dermal exposures to polycyclic aromatic hydrocarbons (PAHs) contained in oil mists. Then, their resultant lung cancer and skin cancer risks were estimated. Finally, control strategies were initiated from the health-risk management aspect. All threading workers in a fastener manufacturing plant were included. 16 inhalatory and 88 dermal PAH exposure samples were collected. Results show that the inhalatory gas phase total PAH exposure level (8.60 x 10(4) ng/m3) was much higher than that of particle phase (2.30 x 10(3) ng/m3). Workers' mean inhalatory exposure level (8.83 x 10(4) ng/m3) was lower, but its corresponding 1-sided upper 95% confidence level (UCL1,95% = 1.02 x 10(5) ng/m3) was higher than the time-weighted average permissible exposure level (PEL-TWA) regulated in Taiwan for PAHs (1.00 x 10(5) ng/m3). The mean whole body total PAHs dermal exposure levels was 5.44 x 10(6) ng/day and the top five exposed surface areas were lower arm, hand, upper arm, neck, and head/front. The estimated lifetime skin cancer risk (9.72 x 10(-3)) was lower than that of lung cancer risk (1.64 x 10(-2)), but both were higher than the significant risk level (10(-3)) defined by the US Supreme Court in 1980. The installation of a local exhaust ventilation system at the threading machine should be considered as the first priority measurement because both lung and skin cancer risks can be reduced simultaneously. If the personal protection equipment would be adopted in the future, both respiratory protection equipment and protective clothing should be used simultaneously. PMID- 18423594 TI - Hh and Wnt signaling regulate formation of olig2+ neurons in the zebrafish cerebellum. AB - The cerebellum, which forms from anterior hindbrain, coordinates motor movements and balance. Sensory input from the periphery is relayed and modulated by cerebellar interneurons, which are organized in layers. The mechanisms that specify the different neurons of the cerebellum and direct its layered organization remain poorly understood. Drawing from investigations of spinal cord, we hypothesized that the embryonic cerebellum is patterned on the dorsoventral axis by opposing morphogens. We tested this using zebrafish. Here we show that expression of olig2, which encodes a bHLH transcription factor, marks a distinct subset of neurons with similarities to eurydendroid neurons, the principal efferent neurons of the teleost cerebellum. In combination with other markers, olig2 reveals a dorsoventral organization of cerebellar neurons in embryos. Disruption of Hedgehog signaling, which patterns the ventral neural tube, produced a two-fold increase in the number of olig2(+) neurons. By contrast, olig2(+) neurons did not develop in embryos deficient for Wnt signaling, which patterns dorsal neural tube, nor did they develop in embryos deficient for both Hedgehog and Wnt signaling. Our data indicate that Hedgehog and Wnt work in opposition across the dorsoventral axis of the cerebellum to regulate formation of olig2(+) neurons. Specifically, we propose that Hedgehog limits the range of Wnt signaling, which is necessary for olig2(+) neuron development. PMID- 18423597 TI - Involvement of NF-kappaB and AP-1 activation in icariin promoted cardiac differentiation of mouse embryonic stem cells. AB - Icariin has been reported to facilitate the differentiation of mouse embryonic stem (ES) cells into cardiomyocytes; however, the mechanism on cardiomyogenic cell lineage differentiation has not been fully elucidated yet. In the present studies, an underlying signaling network including p38, extracellular signal regulated kinase 1, 2 (ERK1, 2), nuclear factor-kappaB (NF-kappaB), activator protein-1 (AP-1) transcription factors c-jun and c-fos was assumed in icariin induced cardiomyogenesis. Icariin rapidly activated p38 and ERK1, 2 in embryoid bodies, treatment with p38 antagonist 4-(4-Fluorophenyl)-2-(4 methylsulfinylphenyl)-5-(4-pyridyl)-1H-imidazole (SB203580) or ERK1, 2 inhibitor 1,4-diamino-2,3-dicyano-1,4-bis[2-aminophenylthio] butadiene (U0126) significantly abolished icariin induced cardiac commitment, MEF2C gene expression and nuclear translocation, as well as cardiac-specific protein alpha-actinin expression, indicating that p38 and ERK1, 2 are specifically involved in icariin stimulated cardiomyogenic cell lineage differentiation of ES cells. Further, IkappaBalpha phosphorylation and NF-kappaB p65 translocation to the nucleus appeared rapidly when embryoid bodies exposed to icariin, and the expression of IkappaBalpha or NF-kappaB p65 in cytoplasm was decreased concomitantly. Moreover, icariin increased c-jun and c-fos mRNA and protein expression. Either SB203580 or U0126 displayed inhibitory effect on icariin induced NF-kappaB and AP-1 activation. It could be concluded that p38 and ERK1, 2 are activated in a coordinated manner, which in turn contribute to NF-kappaB and AP-1 activation in icariin induced cardiomyogenic cell lineage differentiation of mouse ES cells. PMID- 18423598 TI - Protective effect of serofendic acid on ischemic injury induced by occlusion of the middle cerebral artery in rats. AB - We previously reported that a sulfur-containing neuroprotective substance named serofendic acid purified and isolated from fetal calf serum prevented glutamate neurotoxicity in rat cortical cultured neurons. In the present study, we investigated the effect of serofendic acid on ischemic injury induced by a transient occlusion of the middle cerebral artery in rats. Serofendic acid was intracerebroventricularly administered 30 min after the onset of the occlusion. Serofendic acid (30 nmol) significantly reduced total infarct volume, similar to edaravone (30 nmol), a free radical scavenger. Treatment with serofendic acid (1 30 nmol) reduced the infarct volume in a dose-dependent manner. Moreover, serofendic acid (30 nmol) improved neurological deficit scores. These results suggest that intracerebroventricular administration of serofendic acid prevents the neurodegeneration induced by a transient focal cerebral ischemia and reperfusion. PMID- 18423599 TI - Cannabinoid modulation of executive functions. AB - Executive functions are higher-order cognitive processes such as attention, behavioural flexibility, decision-making, inhibitory control, planning, time estimation and working memory that exert top-down control over behaviour. In addition to the role of cannabinoid signaling in other cognitive functions such as mnemonic processes, interest in its involvement in executive functions has arisen more recently. Here, we will briefly review some of the recent findings indicating a modulatory role of cannabinoid action on executive functioning. In addition, a growing body of evidence suggests that in particular adolescents are more vulnerable for the deleterious effects of drugs of abuse such as cannabis on cognitive functioning. Therefore, in this paper we will also briefly discuss some recent developments in this research field. PMID- 18423600 TI - Caveolae and endothelial dysfunction: filling the caves in cardiovascular disease. AB - Discovery in the early 1990s of caveolin-1, the structural protein responsible for maintaining the ohm shape of caveolae, greatly enhanced investigations to elucidate the role of these little caves in the plasma membrane. Perhaps one of the most important realizations concerning caveolae and caveolin is that these elements play an important functional role in the modulation of cell signal transduction pathways, including those involved in endothelial nitric oxide synthase (eNOS) function. Their role was confirmed by studies with caveolin-1 knockout mice which lack caveolae and display abnormal endothelial function responses. One limitation of these knockout models, however, is that absence of the caveolin protein not only results in the lack of caveolae as a structure but also in the lack of interaction/modulation of enzymes/molecules (e.g. eNOS) to which caveolin binds (whether in- or outside caveolae). In contrast to caveolin knockout models, recent experimental findings suggest that in certain cardiovascular diseases caveolin may dissociate from caveolae to the cytosol, hence decreasing the number of caveolae without a change in the total amount of caveolin. Therefore, as the importance of defining the role of caveolins both in caveolae and in cellular regions is being highlighted, it seems also important at the same time to further define the role of caveolae per se being present in the plasma membrane as a structural entity. The objective of this review is to make an explorative tour on the role of caveolae in vascular endothelial function based on existing literature together with some preliminary experimental findings. Evidence and arguments are put forward that alterations in endothelial caveolae do occur in cardiovascular disease and may contribute to the observed endothelial dysfunction in these conditions. PMID- 18423601 TI - Twenty years of dopamine research: individual differences in the response of accumbal dopamine to environmental and pharmacological challenges. AB - Individual differences in the dopaminergic system of the nucleus accumbens of rats have extensively been reported. These individual differences have frequently been used to explain individual differences in response to environmental and pharmacological challenges. Remarkably, only little attention is paid to the factors that underlie these individual differences. This review gives an overview of the studies that have been performed in our institute during the last 20 years to investigate individual differences in accumbal dopamine release. Data are summarised demonstrating that individual differences in accumbal dopamine release are due to individual differences in: the functional reactivity of the noradrenergic system, the accumbal concentration of vesicular monoamine transporters and tyrosine hydroxylase as well as in the quantal size of the presynaptic pools of dopamine. Our data are embedded in the available literature to create a model that illustrates the putative hardware giving rise to the individual-specific release of accumbal dopamine. An important role is contributed to individual differences in the reactivity of the: hypothalamic pituitary-adrenal axes, the reactivity of second messenger systems as well in the aminergic reactivity of the accumbens shell and core. The consequences of the individual-specific make-up and reactivity of the nucleus accumbens on the regulation of behaviour and the response to drugs of abuse will also be discussed. Apart from agents that interact with dopaminergic receptors, re-uptake or breakdown, noradrenergic agents as well as agents that interact with vesicular monoamine transporters or tyrosine hydroxylase are suggested to have therapeutic effects in subjects that are suffering from diseases in which the dopaminergic system is disturbed. PMID- 18423602 TI - Expression and tissue localization of collectin placenta 1 (CL-P1, SRCL) in human tissues. AB - Collectin placenta-1 (CL-P1), also known as scavenger receptor with C-type lectin (SRCL), is a type II membrane glycoprotein that shares structural features with both collectins and type A scavenger receptors. CL-P1 was originally cloned from the placenta and found to be associated with endothelial cells. It binds via its lectin domain to desialyated Lewis X containing glycoproteins and it is able to facilitate internalization of bound ligands. Via positively charged residues in the collagen-like region it binds to negatively charged components of microbial membranes. It has previously been proposed that CL-P1 plays a role in the host defense system and in the clearance of glycoproteins from the blood. With the aims of determining the detailed tissue expression of human CL-P1 we expressed CL P1 recombinantly in both E. coli and CHO cells, and raised monoclonal antibodies against human CL-P1. Three monoclonal antibodies were characterized and used in immunohistochemical analyses of a panel of cryo- and formalin-fixed sections. We find that CL-P1 mainly associates with cytotrophoblasts and syncytiotrophoblasts of the placenta, alveolar macrophages and to a less degree with macrophage-like and stromal cells of the tonsils. By real-time RT-PCR we verified that the placenta is also the main organ of CL-P1 synthesis. The only source of endothelial cells whereto CL-P1 associates are umbilical cord vein endothelial cells (human umbilical vein endothelial cells, HUVEC). In vitro cultured HUVECs express both the CL-P1 mRNA and show anti-CL-P1 immunoreactivity but CL-P1 locates mainly to the cytosol and not to the membrane of these cells. We conclude that CL-P1 is not a common membrane protein on endothelial cells found in normal tissues under steady state conditions. PMID- 18423603 TI - Curcumin induces apoptosis in HCT-116 human colon cancer cells in a p21 independent manner. AB - Several micronutrients present in fruits and vegetables exhibit anticancer activity as a result of their actions on molecular targets involved in carcinogenesis and tumor progression. Curcumin, a phenolic phytochemical derived from the rhizome of Curcuma longa, exhibits both cancer-preventative activity and growth inhibitory effects on neoplastic cells. Several studies report that curcumin inhibits cancer cell proliferation and induces apoptosis in cancer cells through p21-mediated cell cycle arrest. Cancer cells that are deficient in p21 are also reported to be more prone to undergo apoptosis in response to a variety of cytotoxic agents. In this study, we determined whether curcumin-induced cytotoxicity in cultures of HCT-116 human colon cancer cells was dependent on p21 status. Curcumin killed wild-type HCT-116 cells in a dose- and time-dependent manner, as measured in an MTT cell viability assay. Moreover, an equivalent cytotoxic effect by curcumin was observed in both p21(+/+) and p21(-/-)HCT-116 cells, indicating that curcumin-induced cytotoxicity was p21-independent. Primary cultures of human dermal fibroblasts were less sensitive than HCT-116 colon cancer cells to lower doses of curcumin, suggesting a degree of selectivity for neoplastic cells. Western blot analysis showed that cell death in curcumin treated cultures of p21(+/+) and p21(-/-) HCT-116 cells was associated with a reduction in pro-caspase-3 and PARP-1 cleavage, which are indicative of apoptosis. We conclude that curcumin-induced apoptosis in HCT-116 colon cancer cells does not depend on p21 status. PMID- 18423604 TI - Preclinical assessment of drug-induced proarrhythmias: role of the arterially perfused rabbit left ventricular wedge preparation. AB - Drug-induced torsade de pointes (TdP) is a rare but lethal side effect of many cardiovascular and non-cardiovascular drugs. It has led to black box warnings or even withdrawal of many useful compounds from the market and is one of the major stumbling blocks for new drug development. The critical need for a better test that can predict the TdP liability of a candidate drug has led to the development of multiple preclinical models. Each of these models has it own merits and limitations in preclinical testing for TdP liability; however, most of these models have not been adequately validated, so their precise sensitivity and specificity remain largely unknown. Recent blinded validation studies have demonstrated that the rabbit left ventricular wedge preparation can predict drug induced TdP with an extremely high sensitivity and specificity. As a matter of fact, the wedge technique was initially developed primarily for studying the electrical heterogeneity of myocardium and the cellular basis of QT prolongation and TdP. Naturally then, the electrophysiological data obtained from the wedge takes into account every critical factor associated with the development of TdP. The TdP scores generated using the wedge technique have been shown to assess the torsadogenic potential of the drugs in a predictable fashion. This review elaborates on the current and prospective role of the rabbit left ventricular wedge preparation in preclinical assessment of drug-induced proarrhythmias including but not limited to TdP. PMID- 18423605 TI - Growth of cancer cell lines under stem cell-like conditions has the potential to unveil therapeutic targets. AB - Malignant tumors comprise a small proportion of cancer-initiating cells (CIC), capable of sustaining tumor formation and growth. CIC are the main potential target for anticancer therapy. However, the identification of molecular therapeutic targets in CIC isolated from primary tumors is an extremely difficult task. Here, we show that after years of passaging under differentiating conditions, glioblastoma, mammary carcinoma, and melanoma cell lines contained a fraction of cells capable of forming spheroids upon in vitro growth under stem cell-like conditions. We found an increased expression of surface markers associated with the stem cell phenotype and of oncogenes in cell lines and clones cultured as spheroids vs. adherent cultures. Also, spheroid-forming cells displayed increased tumorigenicity and an altered pattern of chemosensitivity. Interestingly, also from single retrovirally marked clones, it was possible to isolate cells able to grow as spheroids and associated with increased tumorigenicity. Our findings indicate that short-term selection and propagation of CIC as spheroid cultures from established cancer cell lines, coupled with gene expression profiling, represents a suitable tool to study and therapeutically target CIC: the notion of which genes have been down-regulated during growth under differentiating conditions will help find CIC-associated therapeutic targets. PMID- 18423607 TI - Errorless learning and memory performance in schizophrenia. AB - There is evidence that patients with schizophrenia have impaired explicit memory and intact implicit memory. The present study sought to replicate and extend that of O'Carroll et al. [O'Carroll, R.E., Russell, H.H., Lawrie, S.M. and Johnstone, E.C., 1999. Errorless learning and the cognitive rehabilitation of memory impaired schizophrenic patients. Psychological Medicine 29, 105-112.] which reported that for memory-impaired patients with schizophrenia performance on a (cued) word recall task is enhanced using errorless learning techniques (in which errors are prevented during learning) compared to errorful learning (the traditional trial-and-error approach). Thirty patients with a DSM-IV diagnosis of schizophrenia and fifteen healthy controls (HC) participated. The Rivermead Behavioural Memory Test was administered and from their scores, the schizophrenic patients were classified as either memory-impaired (MIS), or memory-unimpaired (MUS). During the training phase two lists of words were learned separately, one using the errorless learning approach and the other using an errorful approach. Subjects were then tested for their recall of the words using cued recall. After errorful learning training, performance on word recall for the MIS group was impaired compared to the MUS and HC groups. However, after errorless learning training, no significant differences in performance were found between the three groups. Errorless learning may play an important role in remediation of cognitive deficits for patients with schizophrenia. PMID- 18423606 TI - Aging by epigenetics--a consequence of chromatin damage? AB - Chromatin structure is not fixed. Instead, chromatin is dynamic and is subject to extensive developmental and age-associated remodeling. In some cases, this remodeling appears to counter the aging and age-associated diseases, such as cancer, and extend organismal lifespan. However, stochastic non-deterministic changes in chromatin structure might, over time, also contribute to the break down of nuclear, cell and tissue function, and consequently aging and age associated diseases. PMID- 18423608 TI - Cognitive correlates of schizophrenia signs and symptoms: I. Verbal communication disturbances. AB - We examined the relations between verbal communication disturbances and several hypothesized etiological factors in 47 schizophrenia spectrum individuals. Both alogia and disturbed discourse coherence were associated with poor planning abilities. Alogia and discourse coherence were differentially associated with performance on tasks measuring fluency, working memory, word finding abilities, and concentration/attention. PMID- 18423609 TI - The impact of psychoticism on perceived hassles, depression, hostility, and hopelessness in non-psychiatric African Americans. AB - Psychotic symptoms are distributed along a continuum that extends from normality to diagnosable psychotic disorders and the presence of psychoticism among individuals in the general population may lead to morbidity and social impairment. This study examined a model in which psychoticism leads to several important psychological consequences. The analysis included 134 African Americans with no psychiatric history who were being seen in medical walk-in clinics for non-emergency medical problems. Psychoticism, perceived hassles, depression, hostility, and hopelessness were measured. The Linear Structural Relations Program (LISREL) was used to test the fit of the data to the proposed model, a trimmed hierarchical version, and two alternative models. The data supported a model in which psychoticism has substantial effects on several important characteristics: perceived daily hassles, depression, and hostility. Depression mediated the association between psychoticism and hopelessness. Goodness-of-fit indices for a final trimmed model that eliminated one path from the initial postulated model revealed good fit to the data, and the two alternative models were found not to fit the data. Like psychosis itself, psychoticism appears to cause meaningful dysfunction even among non-psychiatric individuals from the general population. Additional research is needed to further characterize the detrimental effects of psychoticism or self-reported psychotic symptoms in the general, non-psychiatric population. PMID- 18423610 TI - Differentiating cortical patterns of cognitive dysfunction in schizophrenia and posttraumatic stress disorder. AB - Comparative studies are needed to determine whether the cognitive impairments found in various psychiatric disorders are specific to those disorders, or are a more universal consequence of mental illness. This study compares the patterns of cognitive dysfunction in two conditions characterized by working memory dysfunction, schizophrenia and posttraumatic stress disorder (PTSD). Three matched groups (Schizophrenia, PTSD, Control) of 16 subjects had event related potentials recorded, using a 27 electrode array, while they performed a working memory auditory target detection task. Both disorders were associated with impaired task performance, with greater impairment in schizophrenia. Reduction in N1 amplitude was found only in schizophrenia, and an increase in target N2 amplitude and latency was found only in PTSD. Both patient groups showed a reduction in the amplitude of the non-target and target P3, but the groups were distinguished by a reduction in non-target parietal P3 amplitude in the schizophrenia group and a reduction in target P3 amplitude over the left posterior parietal region in the PTSD Group. This study demonstrates that there are specific patterns of cognitive dysfunction associated with schizophrenia and with PTSD. PMID- 18423611 TI - Posttraumatic stress disorder, cognitive function and quality of life in patients with schizophrenia. AB - The purpose of the present study was to assess posttraumatic stress disorder (PTSD), cognitive function, and quality of life in patients with schizophrenia who had a self-reported history of trauma exposure. Outpatients diagnosed with schizophrenia or schizoaffective disorder were referred to the study. Each patient was assessed with the Positive and Negative Syndrome Scale (PANSS), the Harvard Trauma Questionnaire (HTQ), a cognitive assessment battery, Heinrich's Quality of Life Scale (QLS), and the Behavior and Symptom Identification Scale (BASIS). Eighty-seven subjects who reported experiencing at least one traumatic event were included in the study. Fifteen of 87 (17%) met the DSM-IV criteria for PTSD. The PTSD group had significantly worse overall cognitive performance than the non-PTSD group, especially in the domains of attention, working memory and executive function. In addition, the PTSD group showed significantly worse self rated quality of life as measured by the BASIS total score. The development of PTSD is associated with poor cognitive function and subjectively, but not objectively, rated low quality of life in patients with schizophrenia. Evaluating PTSD in patients with schizophrenia could have important implications from both clinical and research perspectives. PMID- 18423612 TI - Schizotypal traits impact upon executive working memory and aspects of IQ. AB - Previous inconsistent findings concerning a link between working memory dysfunction and negative aspects of non-clinical schizotypy have been interpreted to cast doubt on the continuity model of 'negative psychosis-proneness'. This study employed the Letter-Number-Sequencing (LNS) task and the Trail-Making Test to assess more demanding, executive working memory. A secondary concern was to rule out possible mediating effects of familial schizophrenia. It was hypothesised that executive working memory impairment would be associated primarily with negative rather than positive schizotypy even in the absence of familial schizophrenia. Matrix reasoning controlled for IQ. In 87 university student participants with no known family history of schizophrenia, lower LNS scores were associated with higher levels of negative and positive schizotypy traits. Counter to expectations, matrix reasoning scores were also associated with schizotypy, primarily the cognitive/perceptual traits. Results were similar when participants with a known family history of schizophrenia (10) were included (N=97). Findings support the view that impairment of executive working memory (indexed by LNS) is a reliable cognitive marker for negative (and perhaps also positive) schizophrenia vulnerability, independent of familial schizophrenia, and provide the first indication that some facets of IQ (e.g. inductive reasoning) might also be compromised in non-clinical schizotypy. PMID- 18423613 TI - Cognitive correlates of schizophrenia signs and symptoms: II. Emotional disturbances. AB - We examined the cognitive and motor correlates of emotional disturbances in 47 schizophrenia spectrum individuals. Neither affective flattening nor anhedonia was significantly associated with tasks measuring working memory or attention/concentration, or with overall performance on tasks measuring fluency or episodic memory. In contrast, as expected, emotional disturbances were associated with patterns of hemispheric lateralization. Affective flattening and anhedonia were both associated with episodic memory laterality and there were similar trends with motor laterality. Anhedonia was also associated with medication motor side effects. PMID- 18423614 TI - Categorization and evaluation of emotional faces in psychopathic women. AB - Psychopathic individuals have been shown to respond less strongly than normal controls to emotional stimuli. Data about their ability to judge emotional facial expressions are inconsistent and limited to males. To measure categorical and dimensional evaluations of emotional facial expressions in psychopathic and non psychopathic women, 13 female psychopathic forensic inpatients, 15 female non psychopathic forensic inmates and 16 female healthy participants were tested in an emotion-categorizing task. Emotional facial expressions were presented briefly (33 ms) or until buttonpress. Participants were to classify emotional expressions, and to rate their valence and arousal. Group differences in categorization were observed at both presentation times. Psychopathic patients performed worst with briefly presented sad expressions. Moreover, their dimensional evaluation resulted in less positive ratings for happy expressions and less arousal for angry expressions compared with the responses of non psychopathic and normal subjects. Results shed light on the mechanism possibly underlying the emotional deficits in psychopathic women. PMID- 18423615 TI - Sexual assault, mental health, and service use among male and female veterans seen in Veterans Affairs primary care clinics: a multi-site study. AB - This study examined the nature and prevalence of sexual assault (SA), as well as its relationship to psychiatric sequelae and service use, among the veteran population. We performed a secondary data analysis of a cross-sectional dataset consisting of 643 male and 173 female veterans seen in four Veterans Affairs (VA) primary care clinics. Original data were obtained through semi-structured clinic assessments, structured telephone interviews, and medical chart reviews. Analyses included descriptive statistics, chi-square, analysis of variance (ANOVA), and logistic regression. The lifetime prevalence of SA was 38% among women and 6% among men. Of veterans reporting a history of SA, most experienced child sexual abuse and sexual revictimization. SA victims also had a more extensive trauma history and demonstrated greater psychological impairment in comparison to veterans reporting other types of trauma. However, only 25% of male SA survivors and 38% of female SA survivors used mental health services in the past year. These findings suggest that VA primary care clinics may benefit from expanding the current mandated screen for military sexual trauma to include lifetime experiences and trauma-related symptoms, thereby connecting more veterans with needed mental health services. PMID- 18423616 TI - Self-reporting software for bipolar disorder: validation of ChronoRecord by patients with mania. AB - With the widespread recognition of the value of active patient participation in their care, ChronoRecord software was developed to automate daily self-reporting by patients with bipolar disorder. A prior study demonstrated concurrent validity between self-ratings on ChronoRecord and clinician ratings on the Hamilton Depression Rating Scale (HAMD), but validity with the Young Mania Rating Scale (YMRS) could not be shown due to a lack of data when the outpatients were manic (Bauer et al., Bipolar Disorders 6, 67-74, 2004). This study expanded upon the prior validation study to include inpatients with mania. Self-reported mood ratings on ChronoRecord and clinician ratings on the YMRS were obtained on the same day from 27 inpatients (57 ratings); these data were also combined with the ratings from the 80 outpatients (total 107 patients, 340 ratings). Using Pearson correlation, the self-reported ratings on ChronoRecord were significantly correlated with the YMRS. The accuracy of ChronoRecord to discriminate hypomania and mania was high, as described by the area under the receiver operating characteristic curve. Post-hoc analysis of the level of agreement between ChronoRecord and YMRS ratings was excellent or good in all cases using the kappa statistic. These data demonstrate concurrent validity between ChronoRecord and YMRS. PMID- 18423617 TI - Blinded, multi-center validation of EEG and rating scales in identifying ADHD within a clinical sample. AB - Previous validation studies of attention deficit/hyperactivity disorder (ADHD) assessment by rating scales or EEG have provided Class-IV evidence per standards of the American Academy of Neurology. To investigate clinical applications, we collected Class-I evidence, namely from a blinded, prospective, multi-center study of a representative clinical sample categorized with a clinical standard. Participating males (101) and females (58) aged 6 to 18 had presented to one of four psychiatric and pediatric clinics because of the suspected presence of attention and behavior problems. DSM-IV diagnosis was performed by clinicians assisted with a semi-structured clinical interview. EEG (theta/beta ratio) and ratings scales (Conners Rating Scales-Revised and ADHD Rating Scales-IV) were collected separately in a blinded protocol. ADHD prevalence in the clinical sample was 61%, whereas the remainder had other childhood/adolescent disorders or no diagnosis. Comorbidities were observed in 66% of ADHD patients and included mood, anxiety, disruptive, and learning disorders at rates similar to previous findings. EEG identified ADHD with 87% sensitivity and 94% specificity. Rating scales provided sensitivity of 38-79% and specificity of 13-61%. While parent or teacher identification of ADHD by rating scales was reduced in accuracy when applied to a diverse clinical sample, theta/beta ratio changes remained consistent with the clinician's ADHD diagnosis. Because theta/beta ratio changes do not identify comorbidities or alternative diagnoses, the results do not support the use of EEG as a stand-alone diagnostic and should be limited to the interpretation that EEG may complement a clinical evaluation for ADHD. PMID- 18423618 TI - Depressive symptoms induce paranoid symptoms in narcissistic personalities (but not narcissistic symptoms in paranoid personalities). AB - Based on clinical experience, anecdotal reports, and past empirical and conceptual work, we predicted that when people with narcissistic tendencies experience depressive symptoms, they are prone to develop paranoid attitudes. Moreover, we expected that this process was unidirectional, and that the combination of paranoid tendencies and depressive symptoms would not be associated with an increase in narcissistic symptoms. In both cases, results from our 6-month longitudinal study of 71 previously suicidal adults conformed to our expectations. PMID- 18423619 TI - Cognitive correlates of schizophrenia signs and symptoms: III. Hallucinations and delusions. AB - We examined the cognitive correlates of hallucinations and delusions in 47 schizophrenia spectrum individuals. Hallucinations were significantly negatively correlated with performance on episodic memory tasks, and were not significantly associated with performance on tasks measuring fluency or concentration/attention. Although hallucinations were more strongly associated with performance on verbal than non-verbal memory tasks, the difference was not statistically significant. There was also a trend for hallucinations to be associated with poorer performance on working memory tasks, though this association was eliminated when episodic memory performance was taken into account. Delusions were not significantly associated with any of the cognitive measures. PMID- 18423620 TI - Expression and distribution of MMPs and TIMPs in human uveal melanoma. AB - Matrix metalloproteinases (MMPs) and tissue inhibitors of MMPs (TIMPs) are involved in tumour invasion, metastasis and angiogenesis, and have been implicated as progression markers in uveal melanoma, although their topographical expression has not been fully described. In this study we compared the distribution and specificity of several classes of MMPs (MMP-1, -2, -9, -19, and MT1-MMP) and physiological MMP inhibitors (TIMP-2 and -3) in different regions of the tumour microenvironment and adjacent choroid in a series of primary uveal melanomas. Paraffin sections of untreated uveal melanomas (n=18, 3/18 spindle; 11/18 mixed, and 4/18 epithelioid) were examined for MMP-1 (collagenase 1), MMP-2 and MMP-9 (gelatinases A and B), MT1-MMP (membrane-type 1-MMP), MMP-19, TIMP-2 and TIMP-3 (tissue inhibitors of MMPs), using indirect peroxidase immunohistochemistry. The distribution and intensity of immunolabelling was graded semi-quantitatively (0-3) by 2 independent observers. Non-parametric analyses were used to test for associations between tumour cell type, and the average grade of MMP or TIMP expression. Immunostaining for MMP-1, -9, -19 and MT1-MMP was > or =Grade 2 in more than 70% of specimens, and a heterogeneous pattern of MMP-1, -9, MT1-MMP and TIMP-3 expression was observed. At the tumour scleral interface (TSI), melanoma cells had a flattened morphology and a much reduced MMP and TIMP expression, with a high expression in tumour areas adjacent to the TSI. Tumour vasculature and stromal cells strongly expressed MMP-2. We also observed heterogeneous immunostaining of the vasculature by MMP-1, -9, MT1 MMP and TIMP-2 antibodies, and of the extravascular matrix by MMP-9 antibody. The distinct immunostaining patterns observed for MMPs and TIMPs within uveal melanoma are consistent with their involvement in tumour growth and angiogenesis. In particular, the heterogeneous expression within regions of the tumours, and the localized expression in vasculature and stromal cells emphasises the importance of the tumour microenvironment in the pathogenesis of uveal melanoma (and other tumours). PMID- 18423621 TI - Cortical and thalamic components of neocortical kindling-induced epileptogenesis in behaving cats. AB - Kindling is an essential operating paradigm of the nervous system extensively used both as a model of epileptogenesis and neuroplasticity. In a parallel study conducted on chronically implanted non-anesthetized kindled cats, we report the occurrence of long-lasting slow oscillatory patterns (1.5-2 Hz) called outlasting activities (OA) following the acute seizures (AS) induced by cortical stimulation. Here, we asked if OA observed in the neocortex of kindled animals are generated exclusively by the cortical networks or if they also rely on the burst firing of thalamic neurons. We analyzed the electrophysiological patterns of synchronization of cortical EEG (areas 4, 5, 7, 21, 17, 18, 22) and thalamic field (EThG) (ventral posterior lateral nucleus-VPL), and the influence of modulatory systems originating in the pedunculo-pontine tegmentum (PPT) and locus coeruleus (LC) on the discharge pattern of thalamic neurons during OA. Synchrony analysis of field recordings showed that during AS cortical paroxysmal activities preceded thalamic ones, while during OA this sequential order was reversed. During OA thalamic neurons regularly discharged bursts with the frequency of OA. Electrical stimulation of either PPT or LC during OA decreased both the probability of bursts in thalamocortical neurons and the amplitude of OA. Yet, neither of them was able to block completely the expression of OA. Following PPT/LC stimulation the burst firing of thalamocortical neurons was replaced by tonic firing. We conclude that the thalamus is involved in the mechanism of generation of OA but that it does not play an exclusive role. PMID- 18423622 TI - Protein kinase A modulates retinal ganglion cell growth during development. AB - During development, retinal ganglion cells (RGCs) extend their axons toward their thalamic and mesencephalic targets. Their navigation is largely directed by guidance cues present in their environment. Since cAMP is an important second messenger that mediates the neural response to guidance molecules and its intracellular levels seem to decrease significantly following birth, we tested whether modulation of the cAMP/protein kinase A (PKA) pathway would affect the normal development of RGC axons. At postnatal day 1, hamsters received a unilateral intraocular injection of either 0.9% saline solution, 12 mM of the membrane-permeable cAMP analogue (dibutyryl cAMP; db-cAMP), or 10 microM of the PKA inhibitor KT5720. Intraocular elevation of cAMP significantly accelerated RGC axonal growth while inhibition of PKA activity decreased it. Moreover, when highly purified RGC cultures were treated with forskolin (an activator of adenylate cyclase) or cAMP analogues (db-cAMP and Sp-cAMP), neurite length, growth cone (GC) surface area and GC filopodia number were significantly increased. This indicates that intraocular elevation of cAMP acts directly on RGCs. Since these effects were prevented by PKA inhibitors, it demonstrates that cAMP also exerts its action via the PKA pathway. Taken together, these results suggest that the cAMP/PKA cascade is essential for the normal development of retinothalamic projections. PMID- 18423623 TI - An allogeneic hybrid-cell fusion vaccine against canine mammary cancer. AB - Mammary cancer is among the most prevalent of canine tumors frequently resulting in death due to metastatic disease. Most tumors fail to raise an effective immune reaction making improving immune recognition a priority. Hybrid-cell fusion strategies have been employed to load dendritic cell populations with tumor cell antigens to stimulate immune recognition; however, recovery, heterogeneity and quality of primary cells from patients present enormous challenges. We employed allogeneic cell lines to develop an improved hybrid-cell fusion strategy and evaluated immune reactions in normal laboratory beagles. Such a strategy relies on enhanced immune recognition of allogeneic tumor cell antigens by antigen presenting cells. Optimized PEG-promoted fusions between uniquely stained canine mammary tumor CMT12 or CMT28 cells and a dendritic cell-like DH82 cell fusion partner resulted in greater than 40% hybrid-cell fusion populations by flow cytometry and fluorescence microscopy. Hybrid-cell fusions were delivered by direct ultrasound guided injection into popliteal lymph nodes of laboratory beagles. Only hybrid-cell fusions provided statistically significant enhancement of cell-mediated immunity ((51)Cr-release assay) compared to innate reactions in naive vehicle injected dogs while dogs vaccinated with either single cell component alone did not. Vaccination with hybrid-cell fusions enhanced IFN-gamma expression in sorted CD8+ and CD4+ cells but not in CD4-/CD8- cells consistent with a CTL response. Cell-mediated immune assays revealed strong reactions against matched (vaccine component) CMT cells and unmatched CMT cells indicative of an immune response to mammary cancer antigens common to both cell lines. These results provide proof of principle for development of an allogeneic vaccination strategy against canine mammary cancer. PMID- 18423624 TI - The neuropeptide calcitonin gene-related peptide (CGRP) stimulates T cell migration into collagen matrices. AB - CGRP significantly stimulated migration of non-activated and anti-CD3 activated T lymphocytes into a collagen matrix when present inside the collagen, whereas somatostatin-14, NPY, substance P, VIP, beta-endorphin and metenkephalin had no or little effect. The CGRP antagonist CGRP 8-37 abrogated the CGRP-induced cell infiltration. Virtually all migrating cells were CD3+ (>96%) and CGRP did not stimulate B-cell migration. The migration capacity showed no selective relationship to the expression of CD4+, CD8+, CD45RO+ (memory), or CD45RA+ (naive) T cell markers indicating that the regulation of T cell migration is distinct from that of the major T cell phenotypes. PMID- 18423625 TI - Successful pregnancies treated with pioglitazone in infertile patients with polycystic ovary syndrome. AB - OBJECTIVE: To investigate the efficacy of pioglitazone on fecundity in infertile patients with polycystic ovary syndrome (PCOS) who are resistant to conventional ovulation induction such as clomiphene, dexamethasone, or metformin. DESIGN: A retrospective pilot study. SETTING: Department of Obstetrics and Gynecology in a general hospital. PATIENT(S): Nine infertile women with PCOS. INTERVENTION(S): Pioglitazone, 15-30 mg/day, up to 32 weeks. MAIN OUTCOME MEASURE(S): Pregnancy rate. RESULT(S): Seven of nine women became pregnant at an average of 11.3 weeks of initiation of pioglitazone. Four of seven pregnant cases conceived after the first successful induction of ovulation with the initiation of pioglitazone, and two cases conceived after the second cycle. Three women have already delivered, and there is one ongoing pregnancy. The other three cases ended in miscarriage during the sixth or eighth week of pregnancy. CONCLUSION(S): Pioglitazone may be effective in infertile patients with resistant PCOS. PMID- 18423626 TI - High serum concentration of total inhibin in polycystic ovary syndrome. AB - OBJECTIVE: To evaluate serum total inhibin (the sum of precursors, subunits, and mature molecules of inhibin) along with inhibin A and inhibin B in a large sample of women with polycystic ovary syndrome (PCOS) and to investigate whether these parameters differ between lean and overweight PCOS patients. DESIGN: Cross sectional, controlled study. SETTING: Academic health centers in Siena and Bologna, Italy. PATIENT(S): A group of women with PCOS (n = 145) was divided according to body mass index (BMI) into "lean PCOS" (BMI 19-25 kg/m(2), n = 52) and "overweight PCOS" (BMI 26-51 kg/m(2), n = 93). A group of healthy women (n = 90) with BMI 19-24 kg/m(2) and a history of regular menstrual cycles and normal results on physical examination served as controls. INTERVENTION(S): Blood samples were drawn between cycle days 3 and 5 in PCOS patients and controls. MAIN OUTCOME MEASURE(S): Serum concentrations of total inhibin, inhibin A, inhibin B, FSH, and LH were measured using commercially available immunoassays. RESULT(S): Women with PCOS had serum total inhibin levels twice as high as in the control group (median = 115 vs. 47 pg/mL). Dimeric inhibin A concentration was lower in the PCOS group compared with controls (10 vs. 25 pg/mL), whereas inhibin B concentration did not differ significantly between PCOS and control groups. There was no difference of either total inhibin or inhibin A levels between lean and overweight women with PCOS, whereas the inhibin B level was higher in the lean PCOS subgroup compared with the overweight PCOS subgroup. Total inhibin did not show any linear correlation with inhibin B, inhibin A, or serum gonadotropins. CONCLUSION(S): Women with PCOS have high serum concentration of total inhibin but not of inhibin A or inhibin B, thus suggesting that PCOS women have an impaired processing of alpha-inhibin precursor proteins. PMID- 18423627 TI - Aggressive recurrent respiratory papillomatosis in a neonate. AB - Recurrent respiratory papillomatosis is the most common benign laryngeal neoplasm in children. We report an infant with widespread papillomatosis at the time of diagnosis and rapidly progressive disease as a neonate. This child did not have the usual risk factors for recurrent respiratory papillomatosis, being the second child born by cesarean section to a mother without history of condyloma. Repeated surgical debulking has been used to control the disease, along with adjuvant therapies that included systemic therapy with an epidermal growth factor receptor inhibitor and intralesional cidofovir. We report our experience with these therapies. PMID- 18423628 TI - Intragastric pH with oral vs intravenous bolus plus infusion proton-pump inhibitor therapy in patients with bleeding ulcers. AB - BACKGROUND & AIMS: Intravenous bolus plus infusion proton pump inhibitor (PPI) therapy is recommended for patients with bleeding ulcers and higher risk stigmata. If frequent oral dosing of PPIs provided similar antisecretory effect, this might be preferred based on ease and cost. METHODS: Patients presenting with overt bleeding due to ulcers had intragastric pH probes placed after endoscopy and baseline pH recorded. They were randomly assigned to intravenous lansoprazole (90-mg bolus followed by 9-mg/h infusion) or oral lansoprazole (120-mg bolus followed by 30 mg every 3 hours). pH was recorded for 24 hours. RESULTS: Intragastric pH was > 6 for 67.8% of the study period with intravenous PPI (n = 32) and 64.8% with oral PPI (n = 34): difference, 3.0%; 95% confidence interval (CI): -9.2% to 15.2%. Intragastric pH was > 6 for > 60% of the study period in 22 (68.8%) patients receiving intravenous and 22 (64.7%) patients receiving oral PPI: difference, 4.0%; 95% CI: -18.7% to 26.8%. At 1 hour, mean pHs for intravenous vs oral were 5.3 +/- 0.4 vs 3.3 +/- 0.4, respectively (difference, 2.0; 95% CI: 0.8-3.1; P = .001). At > or = 1.5 hours, 95% CIs of the differences for all hourly mean pHs included zero. Mean pH rose above 6 after 2-3 hours of intravenous PPI and 3-4 hours of oral PPI. CONCLUSIONS: Frequent oral PPI may be able to replace the currently recommended intravenous bolus plus infusion PPI therapy in patients with bleeding ulcers, although the possibility that intravenous PPIs are superior cannot be definitively excluded given our relatively wide confidence intervals. Intravenous PPI provides more rapid increase in pH, reaching mean pH of 6 approximately 1 hour sooner than oral PPI. PMID- 18423629 TI - Allosteric inhibitors of chemoattractant receptors: opportunities and pitfalls. AB - Given the central role of chemokines in infection, inflammation and immunity, chemokine receptors are a prime target for pharmacological intervention, and more so after the recent approval of chemokine receptor inhibitors for HIV. Allosteric inhibitors offer a largely unexploited opportunity to interfere with and modulate chemokine receptor activation and signaling. In addition to characterizing binding mode as a first step to understanding the specific mechanism underlying drug action, allosteric inhibitors pose new questions concerning different phases in drug discovery and pharmacological characterization, including the identification of appropriate screening tests, the evaluation of inhibitory effects on different signaling pathways and the implications of agonist- and signaling pathway-dependent inhibition for overall in vivo efficacy. PMID- 18423630 TI - Efficacy and safety of 1-step transnasal endoscopic nasobiliary drainage for the treatment of acute cholangitis in patients with previous endoscopic sphincterotomy (with videos). AB - BACKGROUND: Endoscopic nasobiliary drainage (NBD) for the treatment of acute cholangitis is an accepted method. A recently developed ultrathin transnasal videoendoscope is minimally invasive, even for patients who are critically ill. OBJECTIVE: To evaluate the clinical efficacy and safety of 1-step NBD by transnasal videoendoscopy (TNE). DESIGN: Prospective case study. SETTING: This study was performed at Tokyo Medical University Hospital. PATIENTS: Twenty patients with acute cholangitis who had previously undergone an endoscopic sphincterotomy (ES); including 10 with bile-duct stones, 8 with pancreatic cancers, 1 with chronic pancreatitis, and 1 with benign biliary stricture, were enrolled in this study. An indwelling self-expandable metallic stent (SEMS) was placed in all patients with pancreatic cancers. INTERVENTION: All patients underwent NBD via front-viewing TNE. A 5F NBD catheter was placed into the bile duct. MAIN OUTCOME MEASUREMENT: The efficacy and safety of this technique. RESULTS: The transnasal insertion of TNE was feasible in all patients, and none had epistaxis. Abdominal pain, fever, and jaundice were improved at 24 hours after the procedure in the majority of patients. The mean procedural time was 18.1 minutes. One patient pulled out the NBD catheter. None of the patients died. TNE-NBD was achieved in 19 patients (95%). LIMITATIONS: Maneuverability of the TNE, limited to patients with a previous ES or the placement of an SEMS. CONCLUSIONS: NBD that uses TNE may be a useful and novel technique for the treatment of acute cholangitis in patients with previous ES. PMID- 18423631 TI - Gastroventricular fistula in a patient with Roux-en-Y gastric bypass (with video). PMID- 18423632 TI - A simple method of infecting rabbits with Bovine herpesvirus 1 and 5. AB - This report describes an alternative technique to inoculate rabbits and to reproduce infection by Bovine herpesvirus 1 and 5. First, the nostrils are anaesthetized by aspersion with local anaesthetic. A few seconds later, and after proving the insensitivity of the zone, the rabbits are put on their back legs with their nostrils upwards and the inoculum is introduced slowly into each nostril by using disposable droppers. Clinical signs, viral isolation from nasal swabs, histological lesions found, positive polymerase chain reaction and antibodies production confirm the infection. This very simple and bloodless technique, where the animals are exposed to minor distress, may be useful for evaluating the virulence of BoHV-1 and BoHV-5 strains, to study the establishment of latent virus infection and to test the potential of experimental vaccines or properties of antiviral drugs. It may be also suitable for experimental infection with other respiratory viruses in this animal model. PMID- 18423633 TI - Development of an improved genotyping assay for the detection of hepatitis C virus genotypes and subtypes in Pakistan. AB - A new genotyping system was established for the specific detection of HCV genotypes 1a, 1b, 1c, 2a, 2b, 2c, 3a, 3b, 3c, 4a-h, 5a and 6a during the course of this study. The system is based on entire core region and a part of 5' noncoding region (5'NCR) with genotype-specific primers. Genotype-specific primers were designed on the basis of 114 HCV isolates. Serum samples with known genotypes were used as positive controls to validate the assay developed and to generate PCR band patterns. Band patterns generated from the clinical serum samples from HCV patients were compared to the patterns produced from these control samples. In addition, the type-specific bands were sequenced from the test patients and control clinical samples to validate further the test results. To determine sensitivity and specificity of the assay, a total 260 samples were analyzed simultaneously by this HCV genotyping method and that developed by Ohno and Murex HCV Serotyping 1-6 Assay. The system showed 79.2% concordance with Ohno's system and 65.38% with serotyping system. Samples with discordant results were sequenced and their genotypes were determined by molecular evolutionary analysis. The data indicate that the method described in this study may offer better sensitivity and specificity for the detection directly of HCV genotypes present at low levels in HCV patient samples. PMID- 18423634 TI - Extraction buffer contaminated bacterially as a cause of invalid HIV-1 viral load results on the NucliSens EasyQ system. AB - Shortly after starting to use the NucliSens EasyQ HIV-1 V1.1 system for HIV-1 RNA load testing, the number of invalid tests per assay run gradually increased. Within five days, approximately 50% of tests showed a total lack of amplification of the calibrator and in most cases also of the HIV-1 template. According to the manufacturer's specifications, the lysis buffer and three extraction buffers remain on the automated NucliSens easyMAG extraction system between assay runs. Therefore possible microbial contamination of these buffers was investigated, after they had been on the automated system for approximately one week. The NucliSens easyMAG extraction buffer 2 yielded bacterial growth identified as Acinetobacter baumannii. After regular decontamination of the machine's tubing system with 70% alcohol and storage of the buffers at 4 degrees C between assay runs were commenced, invalid results due to failed internal calibrator signal occurred no longer. It is likely that bacterial contamination of the buffer was the cause of assay failure, probably due to ribonuclease (RNase) activity. Bacterial contamination of PCR systems should be added to the list of potential hazards in diagnostic virology. This experience underlines the necessity of state of-the-art assay design incorporating adequate internal controls and calibrators. PMID- 18423635 TI - Adult hippocampal cell proliferation is suppressed with estrogen withdrawal after a hormone-simulated pregnancy. AB - Estradiol withdrawal after pregnancy is hypothesized to precipitate depressive symptoms in vulnerable women. A hormone-simulated pregnancy was induced in female rats and the effects of a 'postpartum' drop in estradiol on hippocampal cell proliferation were examined. All groups were ovariectomized or given sham surgery prior to treatment. Rats were randomly assigned to 'postpartum', 'postpartum'+EB (estradiol benzoate), 'postpartum'+DPN (diarylpropionitrile; an ERbeta agonist), 'postpartum'+IMI (imipramine; a tricyclic antidepressant), sham, ovariectomized (OVX), sham+IMI or OVX+IMI groups. All 'postpartum' groups received hormone injections (estradiol and progesterone) over 23 days to simulate pregnancy, while IMI groups also received daily imipramine injections. After day 23, 'postpartum' rats were withdrawn from the hormone-simulated pregnancy (mimicking the postpartum drop in gonadal hormones), while other 'postpartum' treatment groups received daily injections of DPN, EB or IMI. On day 3 'postpartum' all rats were injected with bromodeoxyuridine (BrdU; a DNA synthesis marker) and perfused 24 h later to assess cell proliferation and cell death in the dentate gyrus. 'Postpartum' hormone withdrawal decreased hippocampal cell proliferation in the 'postpartum' and 'postpartum'+EB groups only. Chronic imipramine significantly increased hippocampal cell proliferation in sham+IMI, but not OVX+IMI rats suggesting that imipramine's effects to increase hippocampal cell proliferation in female rats is related to reproductive status. Cell death (pyknotic cells) was decreased only in the 'postpartum' group. Together, these results suggest an important, though complex, role for gonadal hormones in the cellular changes accompanying this model of postpartum depression. PMID- 18423636 TI - A panic attack-like unusual stress reaction. AB - Ever since the seminal studies of Hans Selye, activation of hypothalamus pituitary-adrenal (HPA) axis is emblematic of stress. Consequently, the lack of HPA axis responses following the undisputable psychological stress of a panic attack stands out as one of the most intriguing findings of contemporary psychiatry. On the other hand, the defensive behaviors and aversive emotions produced by stimulation of the dorsal periaqueductal gray matter (DPAG) have been proposed as a model of panic attacks. Therefore, we examined whether the plasma levels of 'stress hormones' corticotropin and prolactin show any change following the DPAG-evoked freezing and flight behaviors of the rat. Rats bearing an electrode into the DPAG and an intra-atrial catheter were stimulated at 9:00 a.m., 18-24 h after the catheter implantation. Blood samples were withdrawn just before 1-min stimulation of DPAG, immediately after (5 or 15 min) and throughout 3 to 27 h following stimulation. In another experiment, samples were withdrawn either before or following a prolonged stimulation (5 min) of the DPAG with flight threshold intensity. Hormones were measured by either chemiluminescent or double-antibody immunoassays. Hormone plasma levels following freezing and flight behaviors were compared to those of resting or restraint-stressed rats. Data show that stress hormones remain unaltered following the DPAG-evoked defensive behaviors. Not even the 5-min stimulation of DPAG with the flight threshold intensity changed corticotropin plasma levels significantly. As far as we known, this is the first demonstration of the lack of stress hormone responses following the intense emotional arousal and physical exertion of a fear-like behavior in rats. Data add new evidence of DPAG involvement in spontaneous panic attacks. PMID- 18423637 TI - Sex differences in DHEA and estradiol during development in a wild songbird: Jugular versus brachial plasma. AB - Sexual differentiation of the brain has traditionally been thought to be driven by gonadal hormones, particularly testosterone (T). Recent studies in songbirds and other species have indicated that non-gonadal sex steroids may also be important. For example, dehydroepiandrosterone (DHEA)--a sex steroid precursor that can be synthesized in the adrenal glands and/or brain--can be converted into active sex steroids, such as 17beta-estradiol (E(2)), within the brain. Here, we examine plasma DHEA and E(2) levels in wild developing European starlings (Sturnus vulgaris), from hatch (P0) to fledging (P20). Blood samples were collected from either the brachial vein (n=143) or the jugular vein (n=129). In songbirds, jugular plasma is enriched with neurally-synthesized steroids and, therefore, jugular plasma is an indirect measure of the neural steroidal milieu. Interestingly, brachial DHEA levels were higher in males than females at P4. In contrast, jugular DHEA levels were higher in females than males at P0 and P10. Brachial E(2) levels were higher in males than females at P6. Surprisingly, jugular E(2) levels were not high and showed no sex differences. Also, we calculated the difference between brachial and jugular steroid levels. At several ages, jugular steroid levels were lower than brachial levels, particularly in males, suggesting greater neural metabolism of circulating DHEA and E(2) in males than females. At a few ages, jugular steroid levels were higher than brachial levels, suggesting neural secretion of DHEA or E(2) into the general circulation. Taken together, these data suggest that DHEA may play a role in brain sexual differentiation in songbirds. PMID- 18423638 TI - Social status and cortisol levels in singing rock hyraxes. AB - Many mammals use acoustic signals to communicate with conspecifics. Rock hyraxes (Procavia capensis) are social mammals whose vocal communication is usually restricted to quiet sounds used between nearby individuals. Loud repetitive warning trills are an exception. In our study site, a third of the adult male hyraxes also produces a rich, complex and loud vocalization we term 'singing'. In this study, we examine whether singers, which are more conspicuous by the act of singing, have higher cortisol (i.e. basal stress; C) levels than non-singers, and whether there is an association between social status and stress hormones in male hyraxes. We show that 'singing' males are different from the general adult male population in that their C levels are higher than those of silent males. Only in singers, C levels are associated with social rank, with dominants showing the highest levels. Singers are also on average older and more dominant than most other sexually mature non-singing males. Further, they copulate more than non singers, suggesting that singing males may have higher reproductive success. Our results support the 'stress of domination' hypothesis and indicate that in the rock hyrax singing may reflect high competitive ability, designating singers as a distinct class of males, unique in their personal attributes and behavior. PMID- 18423640 TI - In vitro and in vivo anti-hepatitis B virus activities of a plant extract from Geranium carolinianum L. AB - Natural products provide a large reservoir of potentially active agents with anti hepatitis B virus (HBV) activity. We examined the effect of the polyphenolic extract from Geranium carolinianum L. (PPGC) on HBV replication both in vitro and in vivo. In the human HBV-transfected liver cell line HepG(2) 2.2.15, PPGC effectively suppressed the secretion of the HBV antigens in a dose-dependent manner with IC(50) values of 46.85 microg/ml for HBsAg and 65.60 microg/ml for HBeAg at day 9. Consistent with the HBV antigen reduction, PPGC (100 microg/ml) also reduced HBV DNA level by 35.9%. In the duck hepatitis B virus (DHBV) infected ducks, after PPGC was dosed intragastricly (i.g.) once a day for 10 days, the plasma DHBV DNA level was reduced, with an ED(50) value of 47.54 mg/kg. In addition, Southern blot analysis confirmed the in vivo anti-HBV effect of PPGC in ducks and PPGC also reduced the plasma and the liver DHBV DNA level in a dose dependent manner. Furthermore, significant improvement of the liver was observed after PPGC treatment, as evaluated by the histopathological analysis. PMID- 18423639 TI - Is the anti-psychotic, 10-(3-(dimethylamino)propyl)phenothiazine (promazine), a potential drug with which to treat SARS infections? Lack of efficacy of promazine on SARS-CoV replication in a mouse model. AB - Phenothiazine and derivatives were tested for inhibition of SARS-CoV replication. Phenothiazine slightly inhibited SARS-CoV replication in a neutral red (NR) uptake assay. Adding a propylamino group to give promazine reduced virus yields (VYR assay) with an EC(90)=8.3+/-2.8 microM, but without selectivity. Various substitutions in the basic phenothiazine structure did not promote efficacy. Phenazine ethosulfate was the most potent compound by VYR assay (EC(90)=6.1+/-4.3 microM). All compounds were toxic (IC(50)=6.6-74.5 microM) except for phenoxathiin (IC(50)=858+/-208 microM) and 10-(alpha-diethylamino-propionyl) phenothiazine.HCl (IC(50)=195+/-71.2 microM). Consequently, none were selective inhibitors of SARS-CoV replication (SI values <1-3.3 microM). These data portended the poor efficacy of promazine in a SARS-CoV mouse lung replication model. Intraperitoneal treatment with promazine using a prophylactic ( 4h)/therapeutic regimen of 1, 10, or 50mg/(kg day) did not reduce virus lung titers at day 3, yet prolonged virus replication to 14 days. Similar therapeutic promazine doses were not efficacious. Thus, promazine did not affect SARS-CoV replication in vitro or in vivo, nor were any other phenothiazines efficacious in reducing virus replication. Therefore, treating SARS infections with compounds like promazine is not warranted. PMID- 18423641 TI - Ethical aspects concerning termination of pregnancy following prenatal diagnosis. PMID- 18423642 TI - Aspects of learned fear related to the hippocampus are sleep-dependent. AB - Reduced sleep interferes with contextual but not cued learned fear, and it was suggested that this selectivity reflects underlying neural substrates. The apparent lack of contextual fear in sleep-deprived animals, however, could be secondary to hyperactivity. Also, changing the parameters of cued conditioning can change the neural pathways involved, such that some types of cued fear might be sensitive to sleep loss. To address these issues, we measured fear expressed with conditioned defecation as well as behavior, and used a trace cued learning paradigm. Using the platform-over-water method, male Sprague-Dawley rats were continuously sleep-deprived for 3 days, or for 20 h/day for 3 days. Animals then underwent fear conditioning, and were tested for learning the next day. Sleep deprived or -restricted animals showed a lack of contextual fear at testing, as conditioned freezing and defecation were minimal. Sleep deprivation also blocked cued fear after trace conditioning. Therefore, reduced sleep impairs contextual learning, and impairs cued learning only when the hippocampus is involved. The data support a model in which sleep loss interferes with hippocampal function while sparing amygdala function. PMID- 18423643 TI - The social zebrafish: behavioral responses to conspecific, heterospecific, and computer animated fish. AB - Zebrafish has been in the forefront of developmental biology and genetics, but only recently has interest in their behavior increased. Zebrafish are small and prolific, which lends this species to high throughput screening applications. A typical feature of zebrafish is its propensity to aggregate in groups, a behavior known as shoaling. Thus, zebrafish has been proposed as a possible model organism appropriate for the analysis of the genetics of vertebrate social behavior. However, shoaling behavior is not well characterized in zebrafish. Here, using a recently developed software application, we first investigate how zebrafish respond to conspecific and heterospecific fish species that differ in coloration and/or shoaling tendencies. We found that zebrafish shoaled with their own species but not with two heterospecific species, one of which was a shoaling the other a non-shoaling species. In addition, we have started the analysis of visual stimuli that zebrafish may utilize to determine whether to shoal with a fish or not. We systematically modified the color, the location, the pattern, and the body shape of computer animated zebrafish images and presented them to experimental zebrafish. The subjects responded differentially to some of these stimuli showing preference for yellow and avoidance of elongated zebrafish images. Our results suggest that computerized stimulus presentation and automated behavioral quantification of zebrafish responses are feasible, which in turn implies that high throughput forward genetic mutation or drug screening will be possible in the analysis of social behavior with this model organism. PMID- 18423644 TI - Interprofessional attitudes amongst undergraduate students in the health professions: a longitudinal questionnaire survey. AB - BACKGROUND: Interprofessional education (IPE) introduced at the beginning of pre registration training for healthcare professionals attempts to prevent the formation of negative interprofessional attitudes which may hamper future interprofessional collaboration. However, the potential for IPE depends, to some extent, on the readiness of healthcare students to learn together. OBJECTIVES: To measure changes in readiness for interprofessional learning, professional identification, and amount of contact between students of different professional groups; and to examine the influence of professional group, student characteristics and an IPE course on these scores over time. DESIGN: Annual longitudinal panel questionnaire survey at four time-points of pre-registration students (n=1683) drawn from eight healthcare groups from three higher education institutions (HEIs) in the UK. RESULTS: The strength of professional identity in all professional groups was high on entry to university but it declined significantly over time for some disciplines. Similarly students' readiness for interprofessional learning was high at entry but declined significantly over time for all groups, with the exception of nursing students. A small but significant positive relationship between professional identity and readiness for interprofessional learning was maintained over time. There was very minimal contact between students from different disciplines during their professional education programme. Students who reported gaining the least from an IPE course suffered the most dramatic drop in their readiness for interprofessional learning in the following and subsequent years; however, these students also had the lowest expectations of an IPE course on entry to their programme of study. CONCLUSION: The findings provide support for introducing IPE at the start of the healthcare students' professional education to capitalise on students' readiness for interprofessional learning and professional identities, which appear to be well formed from the start. However, this study suggests that students who enter with negative attitudes towards interprofessional learning may gain the least from IPE courses and that an unrewarding experience of such courses may further reinforce their negative attitudes. PMID- 18423645 TI - Xenoestrogen exposure and effects in English sole (Parophrys vetulus) from Puget Sound, WA. AB - Vitellogenin, a yolk protein produced in the liver of oviparous animals in response to estrogens, normally occurs only in sexually mature females with developing eggs. However, males can synthesize vitellogenin when exposed to environmental estrogens, making the abnormal production of vitellogenin in male animals a useful biomarker for xenoestrogen exposure. In 1997-2001, as part of the Washington State's Puget Sound Assessment and Monitoring Program, we surveyed English sole from a number of sites for evidence of xenoestrogen exposure, using vitellogenin production in males as an indicator. Significant levels of vitellogenin were found in male fish from several urban sites, with especially high numbers of fish affected in Elliott Bay, along the Seattle Waterfront. Intersex fish were rare, comprising only two fish out of more than 2900 examined. Other ovarian and testicular lesions, including oocyte atresia, were also observed, but their prevalence did not appear to be related to xenoestrogen exposure. However, at the Elliott Bay sites where abnormal vitellogenin production was observed in male sole, the timing of spawning in both male and female English sole appeared altered. Sources of xenoestrogens and types of xenoestrogens present in Elliott Bay are poorly documented, but the compounds are likely associated with industrial discharges, surface runoff, and combined sewer outfalls. PMID- 18423647 TI - Anticrossproducts and cross divisions. AB - This paper defines, in the context of conventional vector algebra, the concept of anticrossproduct and a family of simple operations called cross or vector divisions. It is impossible to solve for a or b the equation axb=c, where a and b are three-dimensional space vectors, and axb is their cross product. However, the problem becomes solvable if some "knowledge about the unknown" (a or b) is available, consisting of one of its components, or the angle it forms with the other operand of the cross product. Independently of the selected reference frame orientation, the known component of a may be parallel to b, or vice versa. The cross divisions provide a compact and insightful symbolic representation of a family of algorithms specifically designed to solve problems of such kind. A generalized algorithm was also defined, incorporating the rules for selecting the appropriate kind of cross division, based on the type of input data. Four examples of practical application were provided, including the computation of the point of application of a force and the angular velocity of a rigid body. The definition and geometrical interpretation of the cross divisions stemmed from the concept of anticrossproduct. The "anticrossproducts of axb" were defined as the infinitely many vectors x(i) such that x(i)xb=axb. PMID- 18423646 TI - Development of GCxGC/TOF-MS metabolomics for use in ecotoxicological studies with invertebrates. AB - The majority of metabolomic studies used in ecotoxicology have implemented (1)H NMR analysis. Despite constant improvement, major limitations of NMR-based techniques include relatively low sensitivity that results in an examination of a limited number of metabolites. An alternative approach is the use of liquid or gas chromatography (GC) for separation of metabolites and mass spectrometry (MS) for their quantification and identification. The objective of our study was to develop a two dimensional GC coupled with time of flight MS (GCxGC/TOF-MS) coupled with multivariate analysis to compare metabolite profiles of Diporeia under different environmental conditions. We compared metabolite profiles between Diporeia collected from Lake Michigan (declining populations) to those residing in Lake Superior (stable populations), and also between Diporeia exposed to a chemical stressor (atrazine) and controls. Overall, 76 and 302 total metabolites were detected from the lake comparison and atrazine studies, respectively. Many of the identified metabolites included fatty acids, amino acids, and hydrocarbons. Furthermore, we observed unique and almost non-overlapping metabolite profiles in both studies. In conclusion, we established the feasibility of using GCxGC/TOF-MS for detecting metabolites as well as developed software to align and merge chromatographic peaks to compare metabolite differences between invertebrate groups sampled under different environmental conditions. This ability to detect unique metabolite profiles under different environmental conditions will increase our undertsanding on the physiological processes and whole-organism reponses occuring as a result of exposure to different environmental stressors. PMID- 18423648 TI - Changes in segmental inertial properties with age. AB - The purpose of this study was to examine how the limb segment inertial parameters vary across the decades from the 1920s to the 1970s. Sixty-six males participated in this study, ranging in age from 20 to 79 years. Pre-screening ensured that all subjects were healthy. The inertial properties of the segments were determined by modeling each segment as series of geometric solids. A multivariate analysis of variance (ANOVA) revealed statistically significant differences between decade age groups for the upper arm, forearm, shank, and thigh (p<0.01). Subsequent ANOVAs revealed statistically significant differences for all the inertial properties for the upper arm, the center of mass location for the forearm, and segment mass for the thigh. Linear regression lines were fit to the data so that each inertial parameter for each segment could be predicted by subject's age, with the slope of this regression line indicating the trend in the data. These trends were statistically significant for all forearm inertial parameters, thigh mass and longitudinal moment of inertia, and forearm center of mass location. The changes for the thigh, upper arm, and forearm were consistent with the changes, which would accompany a change in muscle mass with aging. Resultant joint moments were computed for a set of gait data using inertial properties reflective of the subjects from the age extremes in the study. The resulting differences in the knee and hip moments, young versus old, were all less than 4.5%. PMID- 18423649 TI - A variational constitutive model for soft biological tissues. AB - In this paper, a fully variational constitutive model of soft biological tissues is formulated in the finite strain regime. The model includes Ogden-type hyperelasticity, finite viscosity, deviatoric and volumetric plasticity, rate and microinertia effects. Variational updates are obtained via time discretization and pre-minimization of a suitable objective function with respect to internal variables. Genetic algorithms are used for model parameter identification due to their suitability for non-convex, high dimensional optimization problems. The material behavior predicted by the model is compared to available tests on swine and human brain tissue. The ability of the model to predict a wide range of experimentally observed behavior, including hysteresis, cyclic softening, rate effects, and plastic deformation is demonstrated. PMID- 18423650 TI - Ambulatory blood pressure monitoring (ABPM) in nonagenarians. AB - The objective of the study is to investigate ambulatory blood pressure monitoring (ABPM) in a sample of Spanish nonagenarians. We also analyzed the misdiagnosis of hypertension and investigated blood pressure (BP) control in treated hypertensive nonagenarians. Twenty-four-hour ABPM was undertaken in a group of 42 nonagenarians. The 24-h mean, daytime BP, nighttime BP and heart rate (HR) were extracted from the ABPM. Sociodemographic data, the ability to perform basic daily activities, measured by the Barthel index (BI) or instrumental activities revealed by the Lawton and Brody index (LI), cognition, and comorbidity were evaluated. Thirty-one subjects were receiving antihypertensive drug treatment. Twenty-four hour, daytime and sleeping pressures averaged 130/65, 131/68 and 128/63mmHg, respectively. Seventeen (40.5%) of the 42 patients had a daytime BP of 135/85 or higher. In terms of the BP pattern, 8 (19%) subjects were dippers, 19 (45%) non-dippers, and 15 (36%) were risers. Five (45.46%) out of 11 patients with no evidence of hypertension (normotensive patients) had a daytime BP of 135/85 or higher. The mean daytime BP was 135/85 or higher in 12 (38.7%) out of 31 nonagenarians who had previously received therapy for hypertension. In, conclusion a high prevalence of hypertension, misdiagnosis and inadequate BP control was found in nonagenarians treated for hypertension. PMID- 18423651 TI - Determination of organic acids in the presence of inorganic anions by ion chromatography with suppressed conductivity detection. AB - Simultaneous separation of 19 organic acids and 10 inorganic anions has been demonstrated using ion chromatography with a high capacity anion exchange column and the suppressed conductivity detector under an auto-suppression external sulfuric acid mode. Quantitative merits of this method were examined for analysis of nine organic acids of potential significance in cell culture broth. External calibration curves for these analytes were linear with correlation coefficients exceeding 0.999, and the relative standard derivations of observed analyte concentrations were less than 3.0% in both inter- and intra-day evaluations of aqueous standards. Developed methodology was subsequently applied to obtain organic acid profiles of Luria-Bertani liquid media, yeast extract, peptone, and the culture broth of a mutant Escherichia coli strain. Analytes recoveries observed for triplicate analysis of LB media spiked at two concentration levels ranged from 88% to 105% with less than 7% RSD. These data demonstrate quantitative accuracy for LB media and suggest that the report method may also be applicable to complex samples such as fermentation mixture and lignocellulose hydrolysate. PMID- 18423652 TI - Comparison of thin-film microextraction and stir bar sorptive extraction for the analysis of polycyclic aromatic hydrocarbons in aqueous samples with controlled agitation conditions. AB - The characterization of a poly(dimethylsiloxane) (PDMS) thin film for active extraction of some polycyclic aromatic hydrocarbons was investigated in both a 1 L aqueous solution and a flow-through system. The thin film was attached to an electric bench-drill at a constant rotation speed for constant agitation and fast extraction. The performance of a PDMS-coated stir bar at a constant stirring speed was also investigated. The comparison showed that the extraction rate was much higher when using the thin film than the stir bar, and that the equilibration time was greatly shortened due to the larger surface area/volume ratio of the thin film. The extraction rate was roughly proportional to surface area of the extraction phase during the initial stage of the extraction process; the amount of analyte extracted was proportional to the ratio of the extraction phase between the thin film and the stir bar. Different agitation and stirring rates of the thin film and stir bar were applied for extraction, revealing that extraction efficiency can be improved by increasing rotation rate. The application of rotated thin film in the field proved it is a valid and easily operated method for the determination of polycyclic aromatic hydrocarbons (PAHs) in a campus river. However, the stir bar is not as convenient as a thin film in field sampling. PMID- 18423653 TI - Miniaturized dielectric barrier discharge induced chemiluminescence for detection of volatile chlorinated hydrocarbons separated by gas chromatography. AB - A new analytical method was proposed for the detection of volatile chlorinated hydrocarbons (VCHCs) including dichloromethane, 1,2-dichloroethane, chloroform, tetrachloromethane, perfluoromethane and bromoform after gas chromatographic (GC) separation. Atmospheric pressure dielectric barrier discharge (DBD) was miniaturized and used as a GC detector by using chemiluminescence (CL) emission from the reaction of DBD-split VCHCs with luminol solution. The miniaturized DBD CL detector possessed the advantages of simple construction, very low power consumption, and high sensitivity to many volatile halocarbons, especially VCHCs. The parameters of the detector were optimized by using dichloromethane as a typical analyte. Under the optimized experimental conditions, the limit of detection (LOD) down to sub-nmol can be achieved. PMID- 18423654 TI - Simultaneous determination of a spectrum of trichothecene toxins out of residuals of biogas production. AB - A sensitive and selective method for the simultaneous determination of a spectrum of trichothecenes in residuals of biogas production has been developed. It comprises sample clean-up by liquid/liquid partition for digested manure and solid phase extraction for digested solid phase. Quantification of A- and B-type trichothecenes as their trifluoroacetyl derivatives is performed by gas chromatography mass spectrometry (GC/MS), that of B-type trichothecenes alternatively by high-performance liquid chromatography (HPLC). Fluorescence detection (FLD) after post-column derivatisation using methylacetoacetate and ammonium acetate after alkaline decomposition of toxins was applied. Detection limits in digestates were between 1 and 30 microg/l and 20 and 50 microg/l for GC/MS and HPLC/FLD, respectively. Recovery rates were between 52 and 129% for GC/MS detection with the exception of T-2 tetraol with 22%, and between 56 and 123% for HPLC/FLD. PMID- 18423655 TI - Selective gas-phase capture of explosives on metal beta-diketonate polymers. AB - A variety of metal beta-diketonate polymers were assessed for gas-phase selective retention of nitro aromatic, nitrate ester, and peroxide explosives. The La(III) complex of p-di(4,4,5,5,6,6,6-heptafluoro-1,3-hexanedionyl)benzene [La(dihed)] showed 13-42 times the retention for the nitro aromatic compounds compared to a control column (identical column but lacking the 5% loading of the metal beta diketonate polymer). Nitrate esters, the peroxide explosive triacetone triperoxide, and the taggant 1,4-dimethyl-1,4-dinitrobutane were too strongly retained to elute from the La(dihed) column; however, these compounds could be eluted from the less retentive Cu(dihed) or Zn(dihed) columns. A Kovats index of 2124 for 2,4,6-trinitrotoluene (TNT) on the La(dihed) column compared to 1662 on the control illustrates the excellent discrimination against nonpolar hydrocarbons, the principal matrix interference expected in air samples. A proof of-principle experiment demonstrated analysis of an extrapolated 47 part-per trillion (ppt) (v/v) of TNT in an air extract concentrate. PMID- 18423656 TI - Isolation of all-trans lycopene by high-speed counter-current chromatography using a temperature-controlled solvent system. AB - The effect of solvent system, partition coefficient, retention of stationary phase, column, revolution speed, and flow rate of mobile phase are well known parameters to effect HSCCC (high-speed counter-current chromatography) separations. Temperature effects on chromatographic techniques like HPLC and GC are well studied, but the influence of temperature on CCC solvent systems is hardly investigated. This paper presents the influence of temperature on several key parameters (partition coefficient, settling time, volume ratios) in the hydrophobic HSCCC solvent system hexane:dichloromethane:acetonitrile (30:11:18, v/v/v) used for the isolation of lycopene from tomato paste at 10, 15, 20 and 25 degrees C. PMID- 18423657 TI - Does how much and how you pay matter? Evidence from the inpatient rehabilitation care prospective payment system. AB - We use the implementation of a new prospective payment system (PPS) for inpatient rehabilitation facilities (IRFs) to investigate the effect of changes in marginal and average reimbursement on costs. The results show that the IRF PPS led to a significant decline in costs and length of stay. Changes in marginal reimbursement associated with the move from a cost-based system to a PPS led to a 7-11% reduction in costs. The elasticity of costs with respect to average reimbursement ranged from 0.26 to 0.34. Finally, the IRF PPS had little or no impact on mortality or the rate of return to community residence. PMID- 18423658 TI - Structure-activity studies with endogenous allatostatins from Periplaneta americana: expressed receptor compared with functional bioassay. AB - The A-allatostatins (F/YXFGLamides) are insect neuropeptides with inhibitory actions on juvenile hormone (JH) synthesis, muscular contraction and vitellogenesis. They exist in multiple forms within each species. In the cockroach, Periplaneta americana, only one receptor for A-allatostatin has been identified thus far. Here, we have characterised the receptor response to all 15 of the endogenous A-allatostatins encoded by the P. americana allatostatin prohormone gene, together with some analogues, using an indirect heterologous system involving co-expression of the receptor and a potassium channel subunit in Xenopus laevis oocytes and electrophysiological measurements. We have also determined the relative potency of the same peptides to inhibit JH synthesis in corpora allata. Our data reveal that the heterologously expressed receptor responds to all of the endogenous allatostatins and, although differences in potency are recorded, this cannot readily be related to particular differences in the primary structure of the peptides. Similarly, all allatostatins act on the corpora allata to inhibit the synthesis of JH, again with varying potency not readily related to peptide structure. Interestingly, some of the peptides did not perform consistently across the two assays. We show that the receptor is widely expressed in adult P. americana tissues (head, retrocerebral glands, fat body, ovary, male accessory gland, gut, leg muscle, Malpighian tubule and nerve cord) as well as in early larval instars. The spatial expression supports the known pleiotropic activity of allatostatins and role as a paracrine effector. This is the first report of such a detailed characterisation of an invertebrate receptor for allatostatin. PMID- 18423661 TI - Fluorescent labeling of cell-free synthesized proteins with fluorophore conjugated methionylated tRNA derived from in vitro transcribed tRNA. AB - A simple and practical method for preparing fluorophore-conjugated methionylated tRNA necessary for tRNA-mediated fluorescent labeling of cell-free synthesized proteins was developed. Without complicated chromatographic purification and subsequent concentration, fluorophore-conjugated methionylated tRNA with higher purity and fluorescence concentration could be synthesized from in vitro transcribed tRNA instead of from a total tRNA mixture, which has been routinely used as a tRNA source. Although fluorophore-conjugated methionylated tRNA derived from in vitro transcribed tRNA was purified by simple phenol extraction following alcohol precipitation, it worked well in tRNA-mediated fluorescent labeling, yielding an improved signal-to-noise ratio and higher fluorescence intensity compared to the conventional total tRNA-based method. Based on its simplicity in the preparation of labeling agent with higher purity and fluorescence concentration, the developed method will accelerate the prevalence of fluorescence-based detection of cell-free synthesized proteins. PMID- 18423659 TI - Allele and species dependent contractile defects by restrictive and hypertrophic cardiomyopathy-linked troponin I mutants. AB - Restrictive cardiomyopathy (RCM) is a debilitating disease characterized by impaired ventricular filling, reduced ventricular volumes, and severe diastolic dysfunction. Hypertrophic cardiomyopathy (HCM) is characterized by ventricular hypertrophy and heightened risk of premature sudden cardiac death. These cardiomyopathies can result from mutations in the same gene that encodes for cardiac troponin I (cTnI). Acute genetic engineering of adult rat cardiac myocytes was used to ascertain whether primary physiologic outcomes could distinguish between RCM and HCM alleles at the cellular level. Co-transduction of cardiac myocytes with wild-type (WT) cTnI and RCM/HCM linked mutants in cTnI's inhibitory region (IR) demonstrated that WT cTnI preferentially incorporated into the sarcomere over IR mutants. The cTnI IR mutants exhibited minor effects in single myocyte Ca(2+)-activated tension assays yet prolonged relaxation and Ca(2+) decay. In comparison RCM cTnI mutants in the helix-4/C-terminal region demonstrated a) hyper-sensitivity to Ca(2+) under loaded conditions, b) slowed myocyte mechanical relaxation and Ca(2+) transient decay, c) frequency-dependent Ca(2+)-independent diastolic tone, d) heightened myofilament incorporation and e) irreversible cellular contractile defects with acute diltiazem administration. For species comparison, a subset of cTnI mutants were tested in isolated adult rabbit cardiac myocytes. Here, RCM and HCM mutant cTnIs exerted similar effects of slowed myocyte relaxation and Ca(2+) transient decay but did not show variable phenotypes by cTnI region. This study highlights cellular contractile defects by cardiomyopathy mutant cTnIs that are allele and species dependent. The species dependent results in particular raise important issues toward elucidating a unifying mechanistic pathway underlying the inherited cardiomyopathies. PMID- 18423660 TI - Up-regulation of A 2B adenosine receptor in A 2A adenosine receptor knockout mouse coronary artery. AB - In this study, we looked into possible compensatory changes of other adenosine receptors (ARs) in A(2A) genetic knockout mice (A2AKO) as well as the functional role of nitric oxide (NO) in A(2A) AR-mediated vasodilation. Gene expression of ARs from coronary arteries of A(2A) AR wild type mice (A2AWT) and A2AKO was studied using real-time PCR. Functional studies were carried out in isolated heart and isolated coronary artery preparations. A(2B) AR was found to be 4.5 fold higher in A2AKO than in A2AWT, while A(2A) AR expression was absent in A2AKO. There was no difference in A(1) and A(3) ARs between WT and KO animals. The concentration-relaxation curve for adenosine-5'-N-ethylcarboxamide (NECA, non selective AR agonist) in isolated coronary arterial rings in A2AKO was shifted to the left when compared to A2AWT. The concentration-response curve for A(2B) selective agonist (BAY 60-6583) was also shifted to the left in A2AKO hearts. L NAME, a non-specific NO synthase inhibitor, did not affect baseline coronary flow (CF) until the concentration reached 10 microM in A2AWT (76.32+/-11.35% from baseline, n=5). In A2AKO, the CF decreased significantly by L-NAME only at a higher concentration (100 microM, 93.32+/-5.8% from baseline, n=5). L-NMA (1 microM, n=4), another non-specific NO synthase inhibitor, also demonstrated similar results in decreasing CF (59.66+/-3.23% from baseline in A2AWT, while 81.76+/-8.91% in A2AKO). It was further demonstrated that the increase in CF by 100 microM NECA was significantly blunted with 10 microM L-NAME (377.08+/-25.23% to 305.41+/-30.73%, n=9) in A2AWT but not in A2AKO (153.66+/-22.7% to 143.88+/ 36.65%, n=5). Similar results were also found using 50 nM of CGS-21680 instead of NECA in A2AWT (346+/-22.85 to 277+/-31.39, n=6). No change in CF to CGS-21680 was noted in A(2A)AKO. Our data demonstrate, for the first time, that coronary A(2B) AR was up-regulated in mice deficient in A(2A) AR. We also provide direct evidence supporting a role for NO in A(2A) AR-mediated coronary vasodilation. The data further support the role for A(2A) AR in the regulation of basal coronary tone through the release of NO. PMID- 18423662 TI - Detection of Mycobacterium immunogenum by real-time quantitative Taqman PCR. AB - A quantitative real-time 5'-nuclease (Taqman) PCR technique was developed to specifically detect Mycobacterium immunogenum. rpoB-specific primers and Taqman probe were evaluated for detection of M. immunogenum DNA extracted from pure cultures and from industrial metal working fluids (MWFs). Specificity was confirmed and the sensitivity of detection of M. immunogenum genomic DNA was shown to be approximately 9 fg (2 cell equivalents). When tested on industrial metal working fluids from the UK and USA from which no M. immunogenum CFU were recovered, the assay detected between 3.4x10(1) and 1.9x10(4) cell equivalents (CE) per ml, and increased the detection rate over culture to 37.5% (12 of 32 samples). This assay provides a specific, sensitive and rapid method for the detection of M. immunogenum and is applicable within industry for the early detection of this human pathogen and to the possible prevention of hypersensitivity pneumonitis (HP) in workers. PMID- 18423663 TI - The in vivo and in vitro aggregation properties of globular proteins correlate with their conformational stability: the SH3 case. AB - Protein misfolding and deposition underlie an increasing number of debilitating human disorders and constitute a problem of major concern in biotechnology. In the last years, in vitro studies have provided valuable insights into the physicochemical principles underlying protein aggregation. Nevertheless, information about the determinants of protein deposition within the cell is scarce and only a few systematic studies comparing in vitro and in vivo data have been reported. Here, we have used the SH3 domain of alpha-spectrin as a model globular protein in an attempt to understand the relationship between protein aggregation in the test-tube and in the more complex cellular environment. The investigation of the aggregation in Escherichia coli of this domain and a large set of mutants, together with the analysis of their sequential and conformational properties allowed us to evaluate the contribution of different polypeptidic factors to the cellular deposition of globular proteins. The data presented here suggest that the rules that govern in vitro protein aggregation are also valid in in vivo contexts. They also provide relevant insights into intracellular protein deposition in both conformational diseases and recombinant protein production. PMID- 18423664 TI - Characterization of conformational and dynamic properties of natively unfolded human and mouse alpha-synuclein ensembles by NMR: implication for aggregation. AB - Conversion of human alpha-synuclein (aS) from the free soluble state to the insoluble fibrillar state has been implicated in the etiology of Parkinson's disease. Human aS is highly homologous in amino acid sequence to mouse aS, which contains seven substitutions including the A53T that has been linked to familial Parkinson's disease, and including five substitutions in the C-terminal region. It has been shown that the rate of fibrillation is highly dependent on the exact sequence of the protein, and mouse aS is reported to aggregate more rapidly than human aS in vitro. Nuclear magnetic resonance experiments of mouse and human aS at supercooled temperatures (263 K) are used to understand the effect of sequence on conformational fluctuations in the disordered ensembles and to relate these to differences in propensities to aggregate. We show that both aS are natively unfolded at low temperature with different propensities to secondary structure, backbone dynamics and long-range contacts across the protein. Mouse aS exhibits a higher propensity to helical conformation around the C-terminal substitutions as well as the loss of transient long-range contacts from the C- to the N-terminal end and hydrophobic central regions of the protein relative to human aS. Lack of back-folding from the C-terminal end of mouse aS exposes the N-terminal region, which is shown, by (15)N relaxation experiments, to be very restricted in mobility relative to human aS. We propose that the restricted mobility in the N terminal region may arise from transient interchain interactions, suggesting that the N-terminal KTK(E/Q)GV repeats may serve as initiation sites for aggregation in mouse aS. These transient interchain interactions coupled with a non-A beta amyloid component (NAC) region that is both more exposed and has a higher propensity to beta structure may accelerate the rate of fibril formation of aS. PMID- 18423666 TI - Nanotechnology in vascular tissue engineering: from nanoscaffolding towards rapid vessel biofabrication. AB - The existing methods of biofabrication for vascular tissue engineering are still bioreactor-based, extremely expensive, laborious and time consuming and, furthermore, not automated, which would be essential for an economically successful large-scale commercialization. The advances in nanotechnology can bring additional functionality to vascular scaffolds, optimize internal vascular graft surface and even help to direct the differentiation of stem cells into the vascular cell phenotype. The development of rapid nanotechnology-based methods of vascular tissue biofabrication represents one of most important recent technological breakthroughs in vascular tissue engineering because it dramatically accelerates vascular tissue assembly and, importantly, also eliminates the need for a bioreactor-based scaffold cellularization process. PMID- 18423665 TI - Computational studies reveal phosphorylation-dependent changes in the unstructured R domain of CFTR. AB - The cystic fibrosis transmembrane conductance regulator (CFTR) is a cAMP dependent chloride channel that is mutated in cystic fibrosis, an inherited disease of high morbidity and mortality. The phosphorylation of its approximately 200 amino acid R domain by protein kinase A is obligatory for channel gating under normal conditions. The R domain contains more than ten PKA phosphorylation sites. No individual site is essential but phosphorylation of increasing numbers of sites enables progressively greater channel activity. In spite of numerous studies of the role of the R domain in CFTR regulation, its mechanism of action remains largely unknown. This is because neither its structure nor its interactions with other parts of CFTR have been completely elucidated. Studies have shown that the R domain lacks well-defined secondary structural elements and is an intrinsically disordered region of the channel protein. Here, we have analyzed the disorder pattern and employed computational methods to explore low energy conformations of the R domain. The specific disorder and secondary structure patterns detected suggest the presence of molecular recognition elements (MoREs) that may mediate phosphorylation-regulated intra- and inter domain interactions. Simulations were performed to generate an ensemble of accessible R domain conformations. Although the calculated structures may represent more compact conformers than occur in vivo, their secondary structure propensities are consistent with predictions and published experimental data. Equilibrium simulations of a mimic of a phosphorylated R domain showed that it exhibited an increased radius of gyration. In one possible interpretation of these findings, by changing its size, the globally unstructured R domain may act as an entropic spring to perturb the packing of membrane-spanning sequences that constitute the ion permeability pathway and thereby activate channel gating. PMID- 18423667 TI - Effectiveness of lamotrigine in bipolar disorder in a clinical setting. AB - OBJECTIVE: To assess lamotrigine effectiveness in bipolar disorder (BD) patients in a clinical setting. METHOD: Open lamotrigine was naturalistically administered to outpatients at the Stanford University BD Clinic assessed with the Systematic Treatment Enhancement Program for BD (STEP-BD) Affective Disorders Evaluation, and monitored longitudinally with the STEP-BD Clinical Monitoring Form. RESULTS: One hundred and ninety-seven patients (64 BD I, 110 BD II, 21 BD NOS, 2 Schizoaffective Bipolar Type, mean+/-SD age 42.2+/-14.4 years, 62% female) had 200 trials of lamotrigine. Lamotrigine was combined with a mean of 2.1+/-1.5 other psychotropic medications, most often during euthymia or depressive symptoms. Mean lamotrigine duration was 434+/-444 days, and mean final dose was 236+/-132mg/day without valproate, and 169+/-137mg/day with valproate. Lamotrigine was discontinued in only 26.5% of trials at 255+/-242 days, most often due to inefficacy, and seldom due to adverse effects. In 31.5% of trials lamotrigine was continued 264+/-375 days with no subsequent psychotropic added. In 42.0% of trials lamotrigine was continued 674+/-479 days, but had subsequent psychotropic added at 146+/-150 days, most often for anxiety/insomnia and depressive symptoms. In 145 trials started at Stanford, lamotrigine primarily yielded relief of depressive symptoms or maintained euthymia. In 55 trials in which lamotrigine was started prior to Stanford, lamotrigine primarily maintained euthymia. Lamotrigine was generally well tolerated, with no serious rash, and only 3.5% discontinuing due to benign rash. CONCLUSION: In a cohort of bipolar disorder outpatients commonly with comorbid conditions, and most often receiving complex combination therapy, lamotrigine had a low (26.5%, with an overall mean duration of treatment of 434 days) discontinuation rate, suggesting effectiveness in BD in a clinical setting. PMID- 18423668 TI - Gustatory and olfactory sensitivity in patients with anorexia and bulimia in the course of treatment. AB - BACKGROUND: The majority of studies on taste and smell in eating disorders have revealed several alterations of olfactory or gustatory functions. Aim of this prospective study was to employ detailed olfactory and gustatory testing in female subjects of three homogenous groups - anorexia nervosa, bulimia nervosa and healthy controls - and to look at the effects of treatment on these measures. METHODS: Sixteen hospitalized female patients with anorexia (restricting type, mean age [M]=24.5 years), 24 female patients with bulimia (purging type, M=24.3 years) as well as 23 healthy controls (M=24.5 years) received olfactory ("Sniffin' Sticks") and gustatory testing ("Taste Strips"). Group differences in olfactory and gustatory sensitivity, body mass index (BMI), the Beck depression inventory, the eating attitudes test (EAT), and the influence of therapy on gustatory and olfactory function were investigated. RESULTS: (1) Group differences were present for odor discrimination and overall olfactory function with anorexic patients having the lowest scores. (2) Regarding taste function, controls scored higher than patients with anorexia. (3) At admission small but significant correlations were found between overall olfactory function and body weight (r(63)=0.35), BMI (r(63) = 0.37), and EAT score (r(63) = -0.27). Similarly, (4) the taste test score correlated significantly with body weight (r(63)= 0.48), and BMI (r(63) = 0.45). Finally, (5) at discharge overall olfactory and gustatory function were significantly higher compared to admission in anorexic patients. CONCLUSIONS: As compared to healthy controls and bulimic patients our results show lowered olfactory and gustatory sensitivities in anorexic patients that improved with increasing BMI and decreasing eating pathology in the course of treatment. PMID- 18423669 TI - In the mind's eye: provider and patient attitudes on functional brain imaging. AB - Success in functional neuroimaging has brought the promise of quantitative data in the form of brain images to the diagnosis of disorders of the central nervous system for which only qualitative clinical criteria have previously existed. Even though the translation of research to clinical neuroimaging for conditions such as major depression may not be available yet, rapid innovation along this trajectory of discovery to implementation compels exploration of how such information will eventually affect providers and patients. Clinical neuroethics is devoted to elucidating ethical challenges prior to and during the transfer of new research capabilities to the bedside. Through a model of proactive ethics, clinical neuroethics promotes the development of responsible social and public policies in response to new diagnostic and prognostic capabilities for the benefit of patients and their families, and for providers within the health care systems in which they practice. To examine views about the potential interaction of clinical neuroimaging and depression, we surveyed both mental health providers and outpatients and inpatients diagnosed with major depressive disorder. From responses of 52 providers and 72 patients, we found high receptivity to brain scans for treatment tailoring and choice, for improving understanding of and coping with disease, and for mitigating the effects of stigma and self-blame. Our results suggest that, once ready, roll out of the fully validated technology has significant potential to reduce social burden associated with highly stigmatized illnesses like depression. PMID- 18423670 TI - Radiation dose, driving performance, and cognitive function in patients with head and neck cancer. AB - Seven head and neck cancer patients participated in a driving evaluation in a driving simulator. Radiation dose on the temporal lobes was moderately associated with time to complete a cognitive test and with driving performance. Results indicated that incidental irradiation may contribute to a decrease in cognition and in unsafe driving performance, which seems to be time-dependent. PMID- 18423671 TI - A phase II trial of low-dose gemcitabine and radiation alternated to cisplatin and 5-fluorouracil: an active and manageable regimen for stage IV squamous cell carcinoma of the head and neck. AB - BACKGROUND: The addition of gemcitabine may be a reasonable way to enhance the activity of the alternating cisplatin/5-fluorouracil and radiation regimen considered the referring approach for patients with advanced squamous cell carcinoma (SCC) of the head and neck at the National Institute for Cancer Research of Genoa. METHODS: Three courses of cisplatin, 20mg/m(2)/day and 5 fluorouracil, 200mg/m(2)/day, days 1-5 (weeks 1, 4, and 7) alternated to 3 courses of radiotherapy at standard fractionation (weeks 2-3, 5-6, 8-9) up to 60Gy, and gemcitabine, 50mg/m(2) on monday of each week of radiation, were administered to 47 patients with stage IV (42 patients) or relapsed after surgery (5 patients), SCC of the oral cavity, pharynx or larynx. RESULTS: Eighty-five percent of the patients completed the planned treatment. Main grade 3-4 acute toxicities were: mucositis (40%), neutropenia (26%) and thrombocytopenia (30%). Twenty-seven patients reached a complete response (57%). Seven partial responders were rendered disease-free by surgery (final complete response rate: 72%). At a median follow-up of 37 months, 3-year overall survival, progression-free survival and loco-regional control are 50%, 43% and 54%, respectively. CONCLUSIONS: The addition of gemcitabine at low dose to our referring alternating regimen is feasible and very active. It may improve the long-term outcomes despite an acceptable increase of acute mucoseal toxicity. PMID- 18423672 TI - A mathematical model of Ca2+ dynamics in rat mesenteric smooth muscle cell: agonist and NO stimulation. AB - A mathematical model of calcium dynamics in vascular smooth muscle cell (SMC) was developed based on data mostly from rat mesenteric arterioles. The model focuses on (a) the plasma membrane electrophysiology; (b) Ca2+ uptake and release from the sarcoplasmic reticulum (SR); (c) cytosolic balance of Ca2+, Na+, K+, and Cl ions; and (d) IP3 and cGMP formation in response to norepinephrine(NE) and nitric oxide (NO) stimulation. Stimulation with NE induced membrane depolarization and an intracellular Ca2+ ([Ca2+]i) transient followed by a plateau. The plateau concentrations were mostly determined by the activation of voltage-operated Ca2+ channels. NE causes a greater increase in [Ca2+]i than stimulation with KCl to equivalent depolarization. Model simulations suggest that the effect of[Na+]i accumulation on the Na+/Ca2+ exchanger (NCX) can potentially account for this difference.Elevation of [Ca2+]i within a concentration window (150-300 nM) by NE or KCl initiated [Ca2+]i oscillations with a concentration-dependent period. The oscillations were generated by the nonlinear dynamics of Ca2+ release and refilling in the SR. NO repolarized the NE-stimulated SMC and restored low [Ca2+]i mainly through its effect on Ca2+-activated K+ channels. Under certain conditions, Na+-K+-ATPase inhibition can result in the elevation of [Na+]i and the reversal of NCX, increasing resting cytosolic and SR Ca2+ content, as well as reactivity to NE. Blockade of the NCX's reverse mode could eliminate these effects. We conclude that the integration of the selected cellular components yields a mathematical model that reproduces, satisfactorily, some of the established features of SMC physiology. Simulations suggest a potential role of intracellular Na+ in modulating Ca2+ dynamics and provide insights into the mechanisms of SMC constriction, relaxation, and the phenomenon of vasomotion. The model will provide the basis for the development of multi-cellular mathematical models that will investigate microcirculatory function in health and disease. PMID- 18423673 TI - The effect of superspreading on epidemic outbreak size distributions. AB - Recently, evidence has been presented to suggest that there are significant heterogeneities in the transmission of communicable diseases. Here, a stochastic simulation model of an epidemic process that allows for these heterogeneities is used to demonstrate the potentially considerable effect that heterogeneity of transmission will have on epidemic outbreak size distributions. Our simulation results agree well with approximations gained from the theory of branching processes. Outbreak size distributions have previously been used to infer basic epidemiological parameters. We show that if superspreading does occur then such distributions must be interpreted with care. The simulation results are discussed in relation to measles epidemics in isolated populations and in predominantly urban scenarios. The effect of three different disease control policies on outbreak size distributions are shown for varying levels of heterogeneity and disease control effort. PMID- 18423674 TI - A novel method for RNA interference in neurons using enhanced green fluorescent protein (EGFP)-transgenic rats. AB - RNA interference (RNAi) is the simplest way of examining gene function by inhibiting expression. However, due to the low rate of introducing short interfering RNA (siRNA) into neurons, it is difficult to discriminate into which neurons that have been successfully introduced. Here, we used neurons from transgenic rats expressing enhanced green fluorescent protein (EGFP), and we simultaneously applied small interfering RNAs (siRNAs) against EGFP and a targeted gene to the EGFP-expressing neurons. EGFP fluorescence and immunoreactivity of the protein were then assessed by immunofluorescence microscopy. Quantitative analysis of the immunofluorescence confirmed that loss of EGFP closely correlates with loss of the target protein. These results indicate that this method can be used in a wider range of the neuroscientific research, especially in genome-wide studies. PMID- 18423675 TI - Ipsilateral and contralateral serotonergic projections from dorsal and median raphe nuclei to the forebrain in rats: immunofluorescence quantitative analysis. AB - The objective of this study was to clarify the 5-HT projections from the right and left sides of the dorsal (DRD), ventral (DRV) and lateral (DRL) subdivisions of the middle level of the dorsal (DR) raphe nucleus and median (MR) raphe nucleus to the lateral septum (LS), preoptic area (POA) or ventromedial hypothalamus (VMH), which are important neural substrates for neuroendocrine regulation of reproduction. A retrograde neural tracer, Fluoro-Gold (FG), was infused into the right side of these regions in ovariectomized rats and the numbers of FG and/or 5-HT immunopositive cells in the right and left sides of the raphe nuclei were counted. It was found that the POA and VMH received more 5-HT projections than the LS from the DR and MR. In the subdivisions of the DR, 70% of all 5-HT projections from the DR to these 3 areas originated from the DRL. Furthermore, ipsilateral projections from the DR to the POA and VMH but not to the LS were dominant, compared to the contralateral projections. A right-left difference was not seen among the MR 5-HT projections. Thus, laterality of the projections is thought to be strong in the 5-HT clusters located far from the midline of the midbrain raphe nuclei. PMID- 18423676 TI - Long-term outcomes of percutaneous nephrolithotomy compared to shock wave lithotripsy and conservative management. AB - PURPOSE: Percutaneous nephrolithotomy is successful at achieving stone-free status but long-term safety data are lacking. We report our long-term experience with percutaneous nephrolithotomy and compare these results with other treatment modalities. MATERIALS AND METHODS: We identified 87 patients treated with percutaneous nephrolithotomy from 1983 to 1984 who continue to receive care at our institution. Retrospective chart review was performed. Long-term results were compared to those of a group of 288 patients with stones treated nonsurgically (controls) and a group of 288 patients treated with shock wave lithotripsy followed for 19 years. RESULTS: Average followup was 19.2 years (range 12.7 to 23.0). After percutaneous nephrolithotomy new onset renal insufficiency was noted in 9 patients (10.6%), hypertension in 29 (34.1%), diabetes mellitus in 20 (23.5%) and ureteropelvic junction obstruction in 3 (3.5%). Stone recurrence occurred in 32 patients (36.8%). Recurrent stone events were associated with residual fragments after percutaneous nephrolithotomy (p = 0.049). Compared to shock wave lithotripsy there were no significant differences in the development of renal insufficiency, hypertension or diabetes mellitus. Stone recurrence was more common following shock wave lithotripsy (53.5%) compared to percutaneous nephrolithotomy (p = 0.033). Compared to controls there were no significant differences in the development of renal insufficiency or hypertension. On univariate analysis percutaneous nephrolithotomy was associated with the development of diabetes mellitus (p <0.001) but this association did not persist in multivariate analysis. CONCLUSIONS: At 19 years of followup stone recurrences were less frequent following percutaneous nephrolithotomy compared to shock wave lithotripsy. Recurrent stone events were associated with residual fragments after percutaneous nephrolithotomy. Percutaneous nephrolithotomy was not associated with the development of adverse medical conditions compared to shock wave lithotripsy or conservatively managed stone cases. PMID- 18423678 TI - Urinary nerve growth factor level could be a potential biomarker for diagnosis of overactive bladder. AB - PURPOSE: The clinical diagnosis of overactive bladder has great variation and is based on subjective symptoms. This study was designed to measure urinary nerve growth factor levels in patients with different types of overactive bladder and to evaluate whether urinary nerve growth factor could be a biomarker for the diagnosis of overactive bladder. MATERIALS AND METHODS: Urinary nerve growth factor levels were measured in patients with increased bladder sensation, overactive bladder dry and overactive bladder wet, and in a group of control subjects without lower urinary tract symptoms. Measurement of urinary nerve growth factor levels was performed by the enzyme-linked immunosorbent assay. The total urinary nerve growth factor levels were then normalized to the concentration of urinary creatinine (nerve growth factor/creatinine level). Comparison was performed using the nerve growth factor/creatinine level in all subgroups. RESULTS: Urinary nerve growth factor/creatinine levels were low in normal controls (0.041 +/- 0.026) and patients with increased bladder sensation (0.033 +/- 0.02). Patients with overactive bladder dry (0.39 +/- 0.08) or overactive bladder wet (1.7 +/- 0.26) had significantly higher urinary nerve growth factor levels compared to the controls and patients with increased bladder sensation. Patients with overactive bladder wet had significantly higher urinary nerve growth factor levels than those with overactive bladder dry (p = 0.000). The sensitivity of a urinary nerve growth factor/creatinine level of more than 0.05 in the diagnosis of overactive bladder dry or overactive bladder wet was 67.9% and the specificity was 93.8%. CONCLUSIONS: Patients with overactive bladder dry or overactive bladder wet had significantly higher urinary nerve growth factor levels compared to the control group and patients with increased bladder sensation. Urinary nerve growth factor levels could be a potential biomarker for the diagnosis of overactive bladder. PMID- 18423679 TI - Minimally invasive endovascular techniques to treat acute renal hemorrhage. AB - PURPOSE: We evaluated the effectiveness of endovascular therapy for severe renal hemorrhage. MATERIALS AND METHODS: We retrospectively reviewed cases compiled from the trauma database, billing records and interventional radiology logs at our institution from 1990 to 2007. Technical success was defined as the cessation of bleeding after angiographic embolization. Clinical success was defined as the absence of recurrent hematuria without the need for additional embolization. RESULTS: A total of 26 patients underwent angiography and endovascular treatment for renal hemorrhage. Mean patient age was 42 years (median 37, range 7 to 70). There were 20 males and 6 females. Mean clinical followup was 11.7 months. The mechanisms of injury were iatrogenic in 6 cases (renal biopsy in 5 and post percutaneous nephrostomy placement in 1), trauma in 16 (blunt in 10 and penetrating in 6) and spontaneous rupture of a renal mass in 4. At presentation 16 patients (62%) were hemodynamically stable, while 10 (38%) were in shock. A total of 11 patients (42%) presented with gross hematuria, 7 (27%) had microscopic hematuria and 8 (31%) had no evidence of hematuria. A total of 16 patients (62%) had kidney injuries alone, while 10 (38%) also had significant concurrent injuries. Treatment failed in all 5 grade 5 acute renal injuries (100%) caused by external trauma. Technical and clinical success was achieved in 22 (85%) and 17 patients (65%), respectively. CONCLUSIONS: Superselective embolization therapy for renal trauma provides an effective and minimally invasive means to stop bleeding. Overall our complication rate was minimal. Most renal traumas, including most grade 4 injuries, were effectively managed by conservative therapy. Embolization proved effective for grade 4 renal trauma for which conservative therapy failed. In our series embolization failed when applied to grade 5 injuries. PMID- 18423682 TI - Long-term efficacy of distal penile circular fasciocutaneous flaps for single stage reconstruction of complex anterior urethral stricture disease. AB - PURPOSE: We determined the overall efficacy and predictors of success of the distal penile circular fasciocutaneous flap in the management of complex anterior urethral stricture disease not due to lichen sclerosus. MATERIALS AND METHODS: We performed a retrospective review of all patients undergoing reconstruction of complex anterior urethral strictures without lichen sclerosus repaired from 1985 to 2006. Primary and overall stricture-free survival curves were estimated using the Kaplan-Meier method. Cox proportional hazards regression analysis was used to identify univariate and multivariate predictors of flap success. RESULTS: A total of 124 patients met the inclusion and exclusion criteria. Median patient age was 48 years (range 16 to 83). Median followup was 7.3 years (range 1 month to 19.5 years). Median stricture length was 8.2 cm (range 0.5 to 24). At 1, 3, 5 and 10 years the overall estimated stricture-free survival rates were 95%, 89%, 84% and 79%, respectively. On multivariate analysis smoking (HR 4.0, 95% CI 1.2-12.9, p = 0.02), history of hypospadias repair (HR 4.4, 95% CI 1.3-14.6, p = 0.01) and stricture length 7 to 10 cm (HR 7.0, 95% CI 1.4-34.7, p = 0.02) were predictive of failure. CONCLUSIONS: Fasciocutaneous flap urethroplasty has good and durable success rates in the treatment of complex anterior urethral strictures. Predictors of failure included smoking, history of hypospadias repair and longer stricture length. PMID- 18423686 TI - Who will fail local therapy for renal cell carcinoma. PMID- 18423691 TI - Animal models of female stress urinary incontinence. AB - PURPOSE: Urinary incontinence affects 40% of women in the United States and stress urinary incontinence accounts for a large portion of affected patients. As defined by the International Continence Society, stress urinary incontinence is the involuntary leakage of urine upon effort, exertion, sneezing or coughing. Since the ultimate success of long-term management for any condition is based on an understanding of its pathophysiology, and because the pathophysiology of stress urinary incontinence is incompletely defined, animal models have recently been developed to better understand stress urinary incontinence and develop novel treatment alternatives. MATERIALS AND METHODS: Several animal models for urethral dysfunction have emerged in the last few years, including those based on pathophysiological theories of urethral sphincter dysfunction that were designed to simulate maternal birth trauma. Other models have focused on the creation of a durable model of dysfunction for investigating novel treatments. RESULTS: Since animals cannot express intent, these animal models have focused on measuring decreased urethral resistance. The most widely used methods are the sneeze test, the tilt table technique and the leak point pressure test. Newer techniques include abdominal leak point pressure, urethral pressure measurement and retrograde urethral perfusion pressure. In addition to the advantages and disadvantages of each technique, all methods measure the composite contribution to urethral resistance from smooth and striated muscle, urethral closure and connective tissue, although none measures intent. CONCLUSIONS: We critically reviewed the different models of stress urinary incontinence and urethral dysfunction as well as the different methods of measuring urethral resistance. PMID- 18423693 TI - Predicting an optimal outcome after radical prostatectomy: the trifecta nomogram. AB - PURPOSE: The optimal outcome after radical prostatectomy for clinically localized prostate cancer is freedom from biochemical recurrence along with the recovery of continence and erectile function, a so-called trifecta. We evaluated our series of open radical prostatectomy cases to determine the likelihood of this outcome and develop a nomogram predicting the trifecta. MATERIALS AND METHODS: We reviewed the records of patients undergoing open radical prostatectomy for clinical stage T1c-T3a prostate cancer at our center during 2000 to 2006. Men were excluded if they received preoperative hormonal therapy, chemotherapy or radiation therapy, if pretreatment prostate specific antigen was more than 50 ng/ml, or if they were impotent or incontinent before radical prostatectomy. A total of 1,577 men were included in the study. Freedom from biochemical recurrence was defined as post-radical prostatectomy prostate specific antigen less than 0.2 ng/ml. Continence was defined as not having to wear any protective pads. Potency was defined as erection adequate for intercourse upon most attempts with or without phosphodiesterase-5 inhibitor. RESULTS: Mean patient age was 58 years and mean pretreatment prostate specific antigen was 6.4 ng/ml. A trifecta outcome (cancer-free status with recovery of continence and potency) was achieved in 62% of patients. In a nomogram developed to predict the likelihood of the trifecta baseline prostate specific antigen was the major predictive factor. Area under the ROC curve for the nomogram was 0.773 and calibration appeared excellent. CONCLUSIONS: A trifecta (optimal) outcome can be achieved in most men undergoing radical prostatectomy. The nomogram permits patients to estimate preoperatively their likelihood of an optimal outcome after radical prostatectomy. PMID- 18423697 TI - The effect of 5alpha-reductase inhibition with dutasteride and finasteride on bone mineral density, serum lipoproteins, hemoglobin, prostate specific antigen and sexual function in healthy young men. AB - PURPOSE: Dutasteride and finasteride are 5alpha-reductase inhibitors that dramatically decrease serum levels of dihydrotestosterone. Because androgens affect bone, lipids, hematopoiesis, prostate and sexual function, we determined the impact of 5alpha-reductase inhibitors on these end points. MATERIALS AND METHODS: We conducted a randomized, double-blinded, placebo controlled trial of 99 men 18 to 55 years old randomly assigned to receive 0.5 mg dutasteride (33), 5 mg finasteride (34) or placebo (32) daily for 1 year. Bone mineral density was measured at baseline, after 1 year of treatment and 6 months after drug discontinuation. In addition, markers of bone turnover, fasting serum lipoprotein concentrations, hemoglobin and prostate specific antigen were measured at baseline, after 26 and 52 weeks of treatment, and again 24 weeks after drug discontinuation. Sexual function was assessed at these points by a validated questionnaire. RESULTS: Significant suppression of circulating dihydrotestosterone levels with the administration of dutasteride or finasteride did not significantly affect bone mineral density or markers of bone metabolism. Similarly serum lipoproteins and hemoglobin were unaffected. Serum prostate specific antigen and self-assessed sexual function decreased slightly during treatment with both 5alpha-reductase inhibitors but returned to baseline during followup. CONCLUSIONS: Profound suppression of circulating serum dihydrotestosterone induced by 5alpha-reductase inhibitors during 1 year does not adversely impact bone, serum lipoproteins or hemoglobin, and has a minimal, reversible effect on serum prostate specific antigen and sexual function in normal men. Circulating dihydrotestosterone does not appear to have a clinically significant role in modulating bone mass, hematopoiesis or lipid metabolism in normal men. PMID- 18423701 TI - Dual transplantation of marginal kidneys from nonheart beating donors selected using machine perfusion viability criteria. AB - PURPOSE: Viability testing can be used to avoid the transplantation of nonheart beating donor organs that are likely to have primary nonfunction. Such testing also identifies a second group of kidneys which, although unsuitable for solitary transplantation, may be considered for dual transplantation. In kidneys in this group solitary transplants would be unlikely to produce a sufficient glomerular filtration rate to support the recipient. However, if used together as a dual transplant, they have the potential to produce sufficient renal function in 1 patient. MATERIALS AND METHODS: The group at our unit has performed 23 dual nonheart beating donor renal transplants from 2003 to date. Using 3 and 12-month post-transplantation recipient glomerular filtration rates as primary end points we compared our dual transplant group with our series of 115 single nonheart beating donor transplants from 1998 to 2006. RESULTS: At 3 and 12 months mean glomerular filtration rates in the dual group were 46.2 and 45.5 ml per minute per 1.73 m(2), respectively. These values were not significantly different from the mean glomerular filtration rates of 40.7 and 43.0 ml per minute per 1.73 m(2), respectively, in the single transplant group. CONCLUSIONS: We have observed that a subset of nonheart beating donor kidneys that do not satisfy the viability criteria for single organ transplantation may become successful dual organ grafts, thus, avoiding unnecessary organ nonuse and maximizing organ resources. PMID- 18423702 TI - A historical perspective on cystocele repair--from honey to pessaries to anterior colporrhaphy: lessons from the past. AB - PURPOSE: Ancient reports of the treatment of anterior vaginal wall prolapse (cystocele) include the use of honey, astringents and even turning the woman upside down. Various objects were inserted into the vagina to correct this condition. These have since evolved to modern day pessaries. There is limited information on the historical surgical management of cystocele. In this review we provide a historical perspective on the treatment of cystocele. MATERIALS AND METHODS: A MEDLINE search was conducted using the words prolapse, cystocele, etiology, anatomy, pathophysiology, classification systems and the modifications in the nonsurgical and surgical techniques involved in cystocele repair. RESULTS: Development of pelvic surgery finds its roots in the Ebers papyrus (1550 BC) and evolved from Hippocrates (400 BC) who used pessaries with pomegranate to reduce uterine prolapse. Other maneuvers were also used. Vesalius was the first to provide a detailed description of the entire female genital tract. Adolf Retzius defined the boundaries of the prevesical space in 1849. The current concepts regarding the etiology of cystocele were proposed in 1912. Modern pelvic organ surgeons have modified these concepts to popularize new surgical approaches to this ancient clinical problem. CONCLUSIONS: These contributions provide a sound basis for future surgical developments. PMID- 18423703 TI - Ultrasound measurement of bladder wall thickness in the assessment of voiding dysfunction. AB - PURPOSE: Ultrasound measurement of bladder wall thickness has been proposed as a useful diagnostic parameter in patients with bladder outlet obstruction and other voiding dysfunctions. We assessed bladder wall thickness measurement as a noninvasive test in patients with suspected bladder outlet obstruction or overactive bladder syndrome. MATERIALS AND METHODS: Transabdominal ultrasound measurement of bladder wall thickness was performed during urodynamic study in 180 patients with nonneurogenic voiding dysfunction. Two measurements of anterior bladder wall thickness, 1 cm apart in the midline and averaged, were obtained at 200 ml filling. Bladder wall thickness findings were correlated with urodynamic diagnoses. RESULTS: A total of 180 patients with an average age of 62 years (range 20 to 94) were recruited, comprising 73 males and 107 females. Of the patients 69 had normal urodynamics, 39 had bladder outlet obstruction, 38 had increased bladder sensation on cystometry and 34 had detrusor overactivity. Bladder wall thickness was 1.1 to 4.5 mm in all groups. Males had a slightly thicker bladder wall than females (mean 2.1 vs 1.9 mm, p = 0.064). Mean bladder wall thickness in patients with normal urodynamics, bladder outlet obstruction, detrusor overactivity and increased bladder sensation was 2.0, 2.1, 1.9 and 1.8 mm, respectively. No significant difference was found between the groups (ANOVA p = 0.064, not significant). In particular there was no difference in bladder wall thickness between patients with normal urodynamics, and those with bladder outlet obstruction (p = 0.31) or detrusor overactivity (p = 0.309). CONCLUSIONS: Bladder wall thickness is remarkably uniform in patients with nonneurogenic voiding dysfunction. Therefore, it cannot reliably predict bladder outlet obstruction or detrusor overactivity. Bladder wall thickness measurement does not provide an alternative to urodynamic studies for diagnosing voiding dysfunction. PMID- 18423704 TI - Determinants of quality of life for patients with kidney stones. AB - PURPOSE: We evaluated various factors relating to quality of life in a population of patients with stones. MATERIALS AND METHODS: A total of 155 patients seen at our urology clinic for stones between January and May 2007 were prospectively questioned regarding stone history and were administered the SF-36 questionnaire to assess quality of life. Age, body mass index, American Society of Anesthesiologists score, number of stone episodes, interval from the last stone episode, number of surgical procedures for stones and associated complications, missed days of work and long-term medical treatment were elicited from the patients. Individual SF-36 domains and composite scores were compared to those of the American general population. Univariate and multivariate regression analyses were performed to assess the impact of all covariates on quality of life scores. RESULTS: Patients with stones scored lower than the average American population in 5 of the 8 domains of the SF-36 as well as in the physical composite score. Multivariate regression modeling showed that increasing body mass index and age were the strongest predictors of decreased physical well-being. The number of stone episodes did not influence SF-36 scores, although the number of surgical procedures impacted physical and mental components. The number of ureteroscopy procedures and stent placements primarily impacted mental well-being. Medical therapy, particularly the use of potassium citrate, was associated with more favorable quality of life. CONCLUSIONS: Various factors impact quality of life in patients with urolithiasis but the most important are body mass index, age and the number of surgical procedures. Prospective longitudinal studies may further elucidate the determinants of quality of life and they might be used to optimize patient care. PMID- 18423706 TI - Evaluation of semen quality, endocrine profile and hypothalamus-pituitary-testis axis in male patients with homozygous beta-thalassemia major. AB - PURPOSE: The endocrine profile, hypothalamus-pituitary-testis axis and semen quality were evaluated in male patients with homozygous beta-thalassemia major. MATERIALS AND METHODS: A total of 168 male patients 18 years or older with homozygous beta-thalassemia major were enrolled in the study and 84 healthy age matched male volunteers served as controls. All subjects provided a medical history and underwent physical examination and routine semen analysis. Two blood samples were drawn from each participant at 20-minute intervals to determine the resting levels of certain hormones, including luteinizing hormone, follicle stimulating hormone, prolactin, testosterone, estradiol, insulin-like growth factor-1 and insulin like growth factor binding protein-3. The hypothalamus pituitary-testis axis was also assessed using the luteinizing hormone-releasing hormone test. RESULTS: The prevalence of hypogonadotropic hypogonadism was 76.2% (128 patients). Of the studied patients with homozygous beta-thalassemia major 75 (44.6%) were short. Compared with normal controls the patients with homozygous beta-thalassemia major had lower serum luteinizing hormone, follicle-stimulating hormone, insulin-like growth factor-1, insulin-like growth factor-1 binding protein-3 and testosterone. In addition, there were decreased luteinizing hormone and follicle-stimulating hormone responses to luteinizing hormone-releasing hormone in patients with homozygous beta-thalassemia major compared to those in normal controls. Total sperm count, sperm motility and percent normal sperm morphology were lower in patients with homozygous beta-thalassemia major than in controls. CONCLUSIONS: Most subjects with beta-thalassemia major have hypogonadotropic hypogonadism state, impairment fertility and growth retardation. Further studies are warranted to better clarify the causes and consequences of hypogonadotropic hypogonadism in patients with homozygous beta-thalassemia major. PMID- 18423707 TI - Stress urinary incontinence: relative importance of urethral support and urethral closure pressure. AB - PURPOSE: Treatment strategies for stress incontinence are based on the concept that urethral mobility is the predominant causal factor with sphincter function a secondary contributor. To our knowledge the relative importance of these 2 factors has not been assessed in properly controlled studies. MATERIALS AND METHODS: The Research on Stress Incontinence Etiology project is a case-control study that compared 103 women with stress incontinence and 108 asymptomatic controls in groups matched for age, race, parity and hysterectomy. Urethral closure pressure, urethral and pelvic organ support, levator ani muscle function and intravesical pressure were measured and analyzed using logistic regression and multivariable modeling. RESULTS: Mean +/- SD maximal urethral closure pressure was 42% lower in cases (40.8 +/- 17.1 vs 70.2 +/- 22.4 cm H(2)O, d = 1.47). Lesser effect sizes were seen for support parameters, including resting urethral axis and urethrovaginal support (d = 0.41 and 0.50, respectively). Other pelvic floor parameters, including genital hiatus size and urethral axis during muscle contraction (d = 0.60 and 0.58, respectively), differed but levator strength and levator defect status did not. Maximum cough pressure, which is an assessment of stress on the continence mechanism, was also different (d = 0.43). After adjusting for body mass index the maximal urethral closure pressure alone correctly classified 50% of cases. Adding the best predictors for urethrovaginal support and cough strength to the model added 11% of predictive ability. CONCLUSIONS: The finding that maximal urethral closure pressure and not urethral support is the factor most strongly associated with stress incontinence implies that improving urethral function may have therapeutic promise. PMID- 18423709 TI - Patient misunderstanding of the individual questions of the American Urological Association symptom score. AB - PURPOSE: Lower urinary tract symptoms are often assessed using the American Urological Association symptom score. However, some patients may experience difficulty completing the AUA questionnaire. We hypothesized that certain individual questions may generate more misunderstanding than others. MATERIALS AND METHODS: This study involved patients at 2 hospitals who completed the American Urological Association symptom score twice, that is 1) self-administered and 2) physician assisted. Analyses compared self-reported and physician obtained responses to each individual question. One-way ANOVA with the Tukey HSD post hoc test was done to assess whether mean disagreements between self-reported and physician administered American Urological Association symptom scores differed significantly by patient education level. RESULTS: The study group consisted of 998 patients. For each symptom score question we found an inverse relationship between education level and symptom misrepresentation. This discrepancy was the largest for questions on frequency (question 2) and urgency (question 4), which are related to irritative symptoms. Mean misrepresentation of the total American Urological Association symptom score was 2.42 and 5.33 for patients with greater than 12 and fewer than 9 years of education, respectively (p <0.001). Of patients with more than 12 years of education 28% misreported their symptoms by 4 points or greater and 1% misreported them by 10 points or greater, while 58% with fewer than 9 years of education misreported their total score by 4 points or greater and 21% misreported it by greater than 10 points. CONCLUSIONS: While the American Urological Association symptom score is a useful tool for the rapid diagnosis of benign prostatic hyperplasia, patients with low education misrepresent their scores more often and to a higher degree, possibly predisposing them to inappropriate care. PMID- 18423712 TI - Incidence and prevention of iatrogenic urethral injuries. AB - PURPOSE: Although improper urethral catheter insertion is a source of preventable injury in male patients, to our knowledge the extent of this problem is not known. We studied the incidence and mechanism of iatrogenic urinary catheter injuries occurring in adult male inpatients at a single institution. We also designed and implemented an intervention to prevent these injuries. MATERIALS AND METHODS: This study was performed during a 13-month period. During months 1 through 6 all catheter related injuries in all adult male admissions to a single academic tertiary care center were prospectively tracked. Incidence data were calculated, injury severity was analyzed and the injury mechanism was identified. During month 7 based on injury mechanism data a nursing education program was designed and implemented by urology staff that included basic urological anatomy, urethral catheter insertion techniques and catheter safety. During months 8 through 13 catheter related injuries were again tracked. The incidences before and after intervention were compared. RESULTS: During the pre-intervention period iatrogenic urethral catheter injuries occurred in 14 of 4,310 consecutive adult male admissions, representing an incidence of 3.2 injuries per 1,000 patients. Penile and/or perineal pain occurred in all 14 affected patients (100%) and penile bleeding occurred in 12 (86%). One patient required cystoscopy for catheter placement and 1 experienced recurrent urethral strictures requiring multiple dilations. During the post-intervention period a total of 3 injuries occurred in 4,523 consecutive patients, representing a statistically significant decrease in risk by a factor of 4.9 and an incidence rate of 0.7/1,000 adult male admissions (Fisher's exact test p = 0.006). Of the 3 post-intervention injuries 2 (67%) occurred in the operating room, an area that was not targeted as part of the intervention. CONCLUSIONS: Iatrogenic urethral injuries are a substantial source of preventable morbidity in hospitalized male patients. Implementation of a nursing education program may significantly decrease the incidence of iatrogenic urethral injury and, thereby, improve patient safety. PMID- 18423715 TI - Interactive spaced education versus web based modules for teaching urology to medical students: a randomized controlled trial. AB - PURPOSE: This study is an assessment of the acceptability and short-term educational efficacy of interactive spaced education compared to web based teaching cases within the compact time frame of a clinical clerkship. MATERIALS AND METHODS: All 237 third-year students completing their 3-month surgery clerkships at 2 medical schools were asked to complete a urology online-education program covering 4 core topics of benign prostatic hyperplasia, erectile dysfunction, prostate cancer and screening with prostate specific antigen. Students were stratified by clinical site and randomized to 1 of 2 cohorts. Students in cohort A received interactive spaced education on prostate cancer/prostate specific antigen and web based teaching on benign prostatic hyperplasia/erectile dysfunction. Students in cohort B received interactive spaced education on benign prostatic hyperplasia/erectile dysfunction and web based teaching on prostate cancer/prostate specific antigen. A validated 28-item test on all 4 topics was administered at the end of the 10-week program. RESULTS: No statistically significant differences in end-of-program test scores were observed between cohorts in the topics of prostate cancer/prostate specific antigen with 87.6% (SD 12.9) for cohort A (interactive spaced education) and 82.4% (SD 19.6) for cohort B (web based teaching) (p = 0.25). Similarly there was also no statistically significant difference in test scores in the topics of benign prostatic hyperplasia/erectile dysfunction with 79.5% (SD 15.9) for cohort A (web based teaching) and 82.1% (SD 14.7) for cohort B (interactive spaced education, p = 0.28). When students were asked which format they would prefer if they were to receive all their urology online education in a single format, 55% of students (109 of 198 respondents) preferred interactive spaced education while 45% (89 of 109) preferred web based teaching (p = 0.16). CONCLUSIONS: Within the compact time frame of a clinical clerkship interactive spaced education is equivalent to web based teaching in short-term learning gains and in acceptability by medical students. PMID- 18423716 TI - Surgeon experience is strongly associated with biochemical recurrence after radical prostatectomy for all preoperative risk categories. AB - PURPOSE: We have previously reported that there is a learning curve for open radical prostatectomy. In the current study we determined whether the effects of the learning curve are modified by patient risk, as defined by preoperative tumor characteristics. MATERIALS AND METHODS: The study included 7,683 eligible patients with prostate cancer treated with open radical prostatectomy by 1 of 72 surgeons. Surgeon experience was coded as the total prior number of radical prostatectomies done by the surgeon before a patient surgery. Multivariate survival time regression models were used to evaluate the association between surgeon experience and biochemical recurrence separately in each preoperative risk group. RESULTS: We saw no evidence that patient risk affected the learning curve. There was a statistically significant association between biochemical recurrence and surgeon experience on all analyses. The absolute risk difference in a patient receiving treatment from a surgeon with 10 vs 250 prior radical prostatectomies was 6.6% (95% CI 3.4-10.3), 12.0% (95% CI 6.9-18.2) and 9.7% (95% CI 1.2-18.2) in patients at low, medium and high preoperative risk. Recurrence free probability in patients with low risk disease approached 100% for the most experienced surgeons. CONCLUSIONS: Cancer control after radical prostatectomy improves with increasing surgeon experience irrespective of patient risk. Excellent rates of cancer control in patients with low risk disease treated by the most experienced surgeons suggest that the primary reason that recurrence develops in such patients is inadequate surgical technique. The results have significant implications for clinical care. PMID- 18423719 TI - Percutaneous renal cryoablation: experience treating 115 tumors. AB - PURPOSE: We determined technical feasibility, safety and short-term outcomes following percutaneous renal cryoablation. MATERIALS AND METHODS: We performed a retrospective review of 115 renal tumors in 110 patients treated with percutaneous cryoablation. Specific attention was directed to tumor characteristics, hospital course, complications, technical success and treatment success based on followup imaging. RESULTS: Mean tumor size was 3.3 cm (range 1.5 to 7.3), including 29 tumors 4.0 cm or larger and 21 tumors in the anterior kidney. Of 90 renal mass biopsies performed 52 (58%) showed renal cell carcinoma. All patients were admitted to the hospital following cryoablation and most (87%) were discharged home the next day (range 1 to 12 days). There were 7 major complications associated with the 113 cryoablation procedures (6%). Technical success was achieved in 112 of the 115 (97%) treated tumors and 3 residual tumors were seen on 3-month followup imaging. There has been no local progression in 80 tumors (100% treatment success) followed 3 months or longer (mean 13.3 months). CONCLUSIONS: Percutaneous renal cryoablation is technically feasible and relatively safe. With experience many anterior tumors and tumors larger than 4 cm can be successfully treated. Long-term followup remains necessary to prove treatment durability. PMID- 18423723 TI - Absence of viable renal carcinoma in biopsies performed more than 1 year following radio frequency ablation confirms reliability of axial imaging. AB - PURPOSE: Radio frequency ablation is an emerging nephron sparing treatment option in select patients with small renal tumors. Some have questioned the completeness of cell death and the reliability of axial imaging for radio frequency ablation followup. We present results in patients with no evidence of radiographic active disease who underwent biopsy more than 1 year following ablation. MATERIALS AND METHODS: Patients who had no clinical evidence of disease, defined as absent lesion growth and contrast enhancement on computerized tomography, 1 year or more following radio frequency ablation underwent percutaneous renal biopsy to evaluate cell viability in the ablative zone. A total of 19 patients (20 lesions) were included in the study. Histological comparison of pre-ablation and post ablation specimens was performed using hematoxylin and eosin staining. RESULTS: Pre-ablation biopsies confirmed that 17 of 20 tumors were renal cell carcinoma, while the remaining 3 were oncocytoma. Following ablation at a mean followup of 26.9 months (range 13.1 to 58.0) all 20 lesions were stable in size without evidence of contrast enhancement on computerized tomography. At repeat biopsy all histology specimens showed unequivocal tumor eradication with no evidence of cellular viability. Histological changes beyond 1 year demonstrated coagulative necrosis, hyalinization, inflammatory cell infiltration and residual ghost cells. CONCLUSIONS: Pathological examination of radiographically negative lesions biopsied more than 1 year following radio frequency ablation confirmed no evidence of disease in all specimens. Therefore, axial imaging can reliably monitor treatment efficacy in the long term. Chronic changes after radio frequency ablation demonstrate coagulative necrosis and nonviable cells. This suggests an evolution of pathological changes that renders early post-ablative biopsy unreliable. PMID- 18423724 TI - Waiting time from initial urological consultation to nephrectomy for renal cell carcinoma--does it affect survival? AB - PURPOSE: We report survival and recurrence outcomes in all patients undergoing radical or partial nephrectomy for renal cell carcinoma, as related to surgical waiting time. MATERIALS AND METHODS: We retrospectively reviewed the records of 722 patients who underwent surgical resection for renal cell carcinoma. Patients were subdivided by waiting time from the initial urology visit until surgery. Surgical waiting time was evaluated as a continuous variable and by monthly subgroups. Univariate and multivariate analyses were performed to evaluate factors associated with overall, disease specific and recurrence-free survival. RESULTS: Mean time from the first visit to surgery was 1.2 months with 64.1% and 94.3% of patients undergoing surgery within 30 days and within 3 months, respectively. Overall and disease specific survival was not affected by surgical waiting time regardless of how time was analyzed. On univariate analysis 5-year recurrence-free survival was poorer in patients undergoing surgery within 1 month vs more than 1 month (75.7% vs 88.4%, p = 0.02). On multivariate analysis T stage (p <0.0001), grade (p = 0.009), lymph node involvement (p = 0.0001) and histology (p = 0.006) were independent predictors of recurrence-free survival, while surgical waiting time was not (p = 0.18). Surgical waiting time less than 1 month was associated with higher stage and higher grade tumors (p <0.0001 and 0.0006, respectively). CONCLUSIONS: Surgical waiting time from initial urological consultation to operative intervention does not adversely affect the outcome of renal cell carcinoma within the time frames analyzed in this study, in which 94% of cases occurred within 3 months. Individual urologist judgment remains a critical factor in the appropriate and timely care of the patient with a suspicious renal mass. PMID- 18423725 TI - Circulating tumor cells in peripheral blood samples from patients with increased serum prostate specific antigen: initial results in early prostate cancer. AB - PURPOSE: We evaluated the hypothesis that circulating tumor cells as determined using the CellSearch System would correlate with tumor volume, pathological stage and Gleason score in men with localized prostate cancer. MATERIALS AND METHODS: Samples of blood (30 ml) were drawn from 97 men with localized prostate cancer before radical prostatectomy, on postoperative days 2 to 3 and at 6 weeks. A control group consisted of 25 men with an increased prostate specific antigen and no tumor detected on extended prostate biopsy. Samples were analyzed for circulating tumor cells using the CellSearch System. RESULTS: Circulating tumor cells were detected in 21% of patients with cancer and 20% of controls (p = 0.946). At 6 weeks after prostatectomy circulating tumor cells were detected in 16% and 11% (p = 0.51) of the men positive and negative for circulating tumor cells at baseline, respectively. Of the 20 patients with cancer who had circulating tumor cells at baseline 18 showed no circulating tumor cells after surgery. Circulating tumor cell values did not correlate with tumor volume, pathological stage or Gleason score. Only 3.1% of the men with cancer and 8% of the control group had 3 or more circulating tumor cells per 22.5 ml blood at baseline. CONCLUSIONS: In metastatic breast, prostate and other cancers more than 5 circulating tumor cells are often detected using the CellSearch System, and may correlate with prognosis. However, in the setting of localized prostate cancer the number of detectable circulating tumor cells was low, with findings comparable to those in men who were biopsy negative for cancer. We found no correlation between the number of circulating tumor cells and known prognostic factors in this population. PMID- 18423729 TI - Focal prostate therapy. PMID- 18423726 TI - Prolapse and incontinence surgery in older women. AB - PURPOSE: Pelvic floor disorders including urinary incontinence, pelvic organ prolapse and fecal incontinence are common problems encountered by the older woman. With the increasing population of older American women urologists and gynecologists can expect to provide evaluation and treatment of these conditions with increasing frequency. These conditions are amenable to medical and surgical therapies. MATERIALS AND METHODS: Contemporary articles (2003 to the present) that included older women undergoing urogynecological surgery were included in this review. Current data on morbidity, mortality and/or surgical outcomes are presented with evidence based preoperative, intraoperative and postoperative surgical management strategies. RESULTS: Older women undergoing pelvic floor surgery can expect operative risks as well as subjective and objective anatomical and quality-of-life outcomes similar to those of younger women. CONCLUSIONS: The decision for surgical intervention for the treatment of pelvic floor disorders should not be based on chronological age alone. Before selecting a specific surgical procedure, all existing pelvic floor defects should be evaluated. Further research is required to understand the impact that surgery for pelvic floor disorders has on anatomical, physiological and functional outcomes in older women. PMID- 18423732 TI - Barriers to the practice of evidence-based urology. AB - PURPOSE: We investigated urologist perceptions of barriers to implementing evidence-based medicine in clinical practice. MATERIALS AND METHODS: In April 2005 an Internet survey was conducted to assess American Urological Association member attitudes toward evidence-based medicine. This analysis presents the responses to an open-ended question about perceived barriers to implementing evidence-based clinical practice in urology. Two raters developed a coding scheme with 5 main categories of evidence concerns, system level factors, physician factors, patient factors and other barriers. Each rater independently assigned a category to each response. Discrepancies were resolved by consensus. RESULTS: A total of 365 participants (72%) responded to the open-ended question, each providing up to 4 codable responses. Of the group 53% cited concerns about the evidence including the lack thereof, low quality, limited applicability and biased presentation. In addition, 37% reported system level factors such as issues of reimbursement, fear of litigation, problems with implementation, interference/bias by third parties, and expectations and attitudes of other providers as important issues. Physician factors and patient factors were further cited by 28% and 9%, respectively. In terms of the legitimacy of evidence-based medicine 9% expressed skepticism whereas 5% of respondents indicated that they saw no barriers to implementing evidence-based medicine in urology. CONCLUSIONS: Efforts to promote evidence-based medicine in urology should focus not only on the generation of high quality clinical research but also on its unbiased reporting and timely dissemination. Concerted efforts should be made to reduce system level factors that hinder the implementation of evidence-based care. PMID- 18423733 TI - Long-term effects of extracorporeal shock wave lithotripsy on renal function: our experience with 156 patients with solitary kidney. AB - PURPOSE: We studied the long-term impact of shock wave lithotripsy on renal function, stone recurrence and hypertension in patients with a solitary kidney. Patients with a solitary kidney provide a unique opportunity to evaluate any clinically significant change in renal function. MATERIALS AND METHODS: We retrospectively reviewed the records of 156 patients with stones in a solitary kidney treated with shock wave lithotripsy monotherapy. Treatment outcome was evaluated after 3 months. Long-term followup (more than 12 months) was available for 108 patients. Serum creatinine, systolic and diastolic blood pressure, new onset hypertension, calculated glomerular filtration rate, and kidney morphology were determined before and after treatment, and compared by chi-square, paired and unpaired t tests. RESULTS: After 3 months the overall stone-free rate was 80.8% (126 of 156). Renal obstruction caused by steinstrasse after shock wave lithotripsy occurred in 14 (8.9%) patients. Secondary procedures were required in 20 (12.8%) patients. Followup ranged from 1 to 16 years with a mean of 3.8 (SD +/ 3.5). After long-term followup the stone-free rate was 76.8% and real stone recurrence developed in 18.5% of patients. There was no significant difference in any evaluated pretreatment or posttreatment parameters. CONCLUSIONS: The demonstrated effectiveness, small number of complications at short-term followup, insignificant effect on renal function, blood pressure and relatively small number of recurrences at the long-term followup confirm that shock wave lithotripsy is not only effective but is also safe in the long run. PMID- 18423734 TI - An approaching new wave of multicomponent biomarker diagnostics? PMID- 18423735 TI - Preoperative nomogram predicting 12-year probability of metastatic renal cancer. AB - PURPOSE: For patients with renal masses localized to the kidney there is currently no preoperative tool to predict the likelihood of metastatic recurrence following surgical intervention. We developed a predictive model that could be used in the preoperative setting. MATERIALS AND METHODS: We pooled institutional databases from Memorial Sloan-Kettering and Mayo Clinic, and identified complete data on 2,517 patients with renal masses and no concurrent evidence of metastases who underwent radical or partial nephrectomy. Cox proportional hazard regression analyses were used to model preoperative clinical and radiographic characteristics as predictors for development of metastases following nephrectomy. Internal validation was performed with a statistical bootstrapping technique. RESULTS: Metastatic recurrence developed in 340 of the 2,517 patients. Median followup for patients without metastatic recurrence was 4.7 years. A nomogram was developed using preoperative characteristics to predict the 12-year likelihood of postoperative metastatic recurrence with a concordance index of 0.80. In contrast, the concordance index of preoperative TNM staging was 0.71. Size of the primary renal mass, evidence of lymphadenopathy or necrosis on preoperative imaging and the mode of presentation were important predictors for the subsequent development of metastases. CONCLUSIONS: We present a preoperative nomogram that accurately predicts the development of metastatic recurrence following nephrectomy. This nomogram may be potentially useful to identify and counsel patients at high risk for recurrence. PMID- 18423736 TI - Residual tumor potentially left behind after local ablation therapy in prostate adenocarcinoma. AB - PURPOSE: We examined contralateral prostate cancer potentially left behind by focal therapy. MATERIALS AND METHODS: We investigated 100 completely embedded radical prostatectomy specimens in which needle biopsy predicted limited disease (less than 3 positive cores, 50% or less involvement of any positive core, Gleason score 6 or less) and all positive needle cores were unilateral. Clinical stage was T1c in 85 and T2a in 15 cases with the palpable lesion on the positive biopsy side. RESULTS: There was 1 positive core in 66 cases. On average 13.9% of each positive core was involved with tumor. The mean number of separate tumor nodules per radical prostatectomy was 2.9. In 65 radical prostatectomy specimens there was some tumor contralateral to the positive biopsy side. Total tumor volume in the radical prostatectomy contralateral to the positive biopsy side averaged 0.2 cm(3) (largest 1.3). In 23 cases contralateral tumor volume was greater than biopsy positive side tumor volume. There were 13 cases in which more than 0.5 cm(3) cancer was contralateral to the positive biopsy and 7 with predominantly anterior tumor. Volume contralateral to positive biopsy side could not be predicted by the number of positive cores (1 vs 2) or maximum percent of the core involved. Gleason pattern 4, extraprostatic extension or positive margins were seen contralateral to the positive biopsy side in 13, 1 and 2 cases, respectively. CONCLUSIONS: In a highly selected population with limited unilateral biopsy cancer, tumor contralateral to the positive biopsy side at radical prostatectomy is typically small. However, 20% of radical prostatectomy specimens had some contralateral adverse pathology in terms of size, extraprostatic extension, grade or margins. PMID- 18423738 TI - Diagnostic and prognostic molecular markers in renal cell carcinoma. AB - PURPOSE: We reviewed the contemporary literature on molecular biomarkers in renal cell carcinoma and their prognostic significance. MATERIALS AND METHODS: Articles published during 1981 to 2007 in English on renal cell carcinoma were surveyed using the MEDLINE/PubMed database. The subject headings included were genetics, biomarkers, prognosis and risk models of renal cell carcinoma. We present a synthesis of currently known renal cell carcinoma biomarkers at various stages of development and their clinical significance, and prognostic nomograms incorporating biomarkers. RESULTS: The beneficiary role of biomarkers in renal cell carcinoma is challenged by the relatively low prevalence of the disease. Even if a biomarker for renal cell carcinoma had 100% sensitivity and 99.4% specificity, the positive predictive value of the marker in men older than 65 years would be only 10%. Several biomarkers are being investigated in renal cell carcinoma, of which many relate to pathogenic molecular changes that are currently therapeutic targets. Carbonic anhydrase IX is a von Hippel-Lindau mediated enzyme that is expressed in most renal cell carcinoma cases. High (greater than 85%) expression of this marker indicates favorable prognosis and may predict the response to interleukin-2 therapy. B7-H1 expression in renal cell carcinoma cells/lymphocytes may indicate worse survival, possibly through impaired host antitumor immunity. Prognostic nomograms incorporating clinical variables and molecular markers to refine the prediction of treatment outcomes are in active development and await prospective clinical validation. CONCLUSIONS: Several renal cell carcinoma molecular markers appear promising to refine the prognosis and prediction of localized, advanced or metastatic renal cell carcinoma. Currently carbonic anhydrase IX is the best studied and promising marker. Prospective, multicenter clinical validation aimed at the practical clinical usefulness of renal cell carcinoma biomarkers is warranted. PMID- 18423739 TI - Tumor characteristics of carriers and noncarriers of the deCODE 8q24 prostate cancer susceptibility alleles. AB - PURPOSE: In collaboration with deCODE Genetics Inc. we previously reported on the association between genetic variants on chromosome 8q24 and prostate cancer susceptibility. Several prior studies have examined the relationship between these 8q24 alleles and clinical tumor features. In this study we examine the differences in clinical and pathological tumor features between carriers and noncarriers of these 8q24 alleles in patients with prostate cancer. MATERIALS AND METHODS: We genotyped 551 white men who underwent radical prostatectomy or radiation therapy for clinically localized prostate cancer at single institution between 2002 and 2005. Of these men 177 (32.1%) were carriers of the -8 allele of the microsatellite marker DG8S737, the A allele of the single nucleotide polymorphism rs1447295 and/or the A allele of the rs16901979 (a surrogate single nucleotide polymorphism of HapC) 8q24 alleles. We used statistical analyses to compare the distribution of clinical characteristics and pathological outcomes between carriers and noncarriers. RESULTS: The -8, A and HapC surrogate single nucleotide polymorphism alleles were present in 77 (14%), 128 (23%) and 61 (14%) patients with prostate cancer, respectively. Carriers of the -8 or multiple 8q24 alleles were significantly more likely to have a Gleason score of 7 or greater and lymph node metastases. Among men with a family history of prostate cancer, carriers of the -8 allele had a significantly greater risk of high grade disease (64% vs 39%, p = 0.04). CONCLUSIONS: In our predominantly surgically treated population there was a significant association between 8q24 prostate cancer susceptibility alleles, particularly the -8 allele, and high grade disease. In men with a family history of prostate cancer the presence of 1 or more of these alleles also conferred a greater risk of some adverse pathological features. PMID- 18423740 TI - Oncological evaluation of prostate sparing cystectomy: the Montsouris long-term results. AB - PURPOSE: Prostate sparing cystectomy provides an alternative therapeutic option in highly selected patients with invasive bladder cancer who wish to avoid the significant functional side effects of traditional surgery. Concern exists regarding the oncological safety of this technique especially with regard to the presence of prostatic urothelial cancer and incidental prostate adenocarcinoma. We present the long-term oncological outcomes of a large series of patients treated at a single institution. MATERIALS AND METHODS: Between October 1992 and June 2004 a total of 117 patients were selected for prostate sparing cystectomy after meeting the inclusion criteria. RESULTS: Nine patients underwent radical cystoprostatectomy after prostate urothelial carcinoma was diagnosed intraoperatively. Long-term oncological results (2 years minimum followup [mean 55 months]) are presented here for the remaining patients. The rate of local and distant recurrence was 4.7% and 34%, respectively, at 20 months. Overall survival at 5 years in our study was 67%, and 5-year disease-free survival rates were 77% for pT2 N0 or less, 44% for pT3 N0 or greater and 22% for pN+ disease. Of 6 patients found to have prostate adenocarcinoma in transurethral prostate resection specimens 1 was treated with high intensity focused ultrasound and 5 were followed with active surveillance, 2 of whom later died of bladder cancer. CONCLUSIONS: We report oncological data on the largest prospective cohort of patients, with the longest followup, treated by prostate sparing cystectomy to date. Outcomes are comparable with the largest published series of cystoprosatectomies. With appropriate screening the risk of a clinically significant prostate cancer appears to be low. For certain selected patients this technique represents a valuable additional option for treatment. PMID- 18423744 TI - Pharmacoangiographic evidence of the presence and anatomical dominance of accessory pudendal artery(s). AB - PURPOSE: Potency preservation is one of the principal concerns surrounding newer developments in the management of organ confined carcinoma prostate. Nerve sparing techniques may not solely preserve erectile function and it is known that vascular factors may be an etiology of the dysfunction. The role of accessory pudendal arteries in the etiology and prevention of erectile dysfunction after radical prostatectomy is at present unclear. We reviewed pudendal angiograms in patients with erectile dysfunction to evaluate the prevalence and importance of these vessels. MATERIALS AND METHODS: Selective pudendal pharmacoangiograms were obtained in 79 consecutive patients with a history of erectile dysfunction. The aim was to identify accessory pudendal arteries, their origin and their significance relative to all identifiable pudendal arteries and the dorsal penile artery with respect to penile arterial inflow. RESULTS: An accessory pudendal artery was identified in 28 (35%) of the patients. The most common origin was the obturator artery. In 15 of the 28 men (54%) in whom an accessory artery was identified it appeared angiographically to be the dominant penile artery. In 3 patients it was apparently the only major arterial inflow to the penis. CONCLUSIONS: Accessory pudendal arteries may be identifiable with pharmacoangiograms in approximately a third of all men. Because they may be the dominant source of blood supply to the penis in some cases, their preservation during radical prostatectomy could be critical to erectile function following radical prostatectomy. PMID- 18423743 TI - Prostate Specific Antigen Working Group guidelines on prostate specific antigen doubling time. AB - PURPOSE: Prostate specific antigen is a glycoprotein found almost exclusively in normal and neoplastic prostate cells. Prostate specific antigen doubling time, or the change in prostate specific antigen over time, has emerged as a useful predictive marker for assessing disease outcome in patients with prostate cancer. It is important to agree on definitions and values for the calculation of prostate specific antigen doubling time, and to develop a common approach to outcome analysis and reporting. MATERIALS AND METHODS: In September 2006 a conference was held at the National Cancer Institute in Bethesda, Maryland to define these parameters and develop guidelines for their use. RESULTS: The Prostate Specific Antigen Working Group defined criteria regarding prostate specific antigen doubling time including the calculation of prostate specific antigen doubling time, evidence to support prostate specific antigen doubling time as a predictive factor in the setting of biochemical recurrence and the use of prostate specific antigen doubling time as a stratification factor in clinical trials. CONCLUSIONS: We propose that investigators calculate prostate specific antigen doubling time before enrolling patients in clinical studies and calculate it as an additional measurement of therapeutic activity. We believe we have developed practical guidelines for the calculation of prostate specific antigen doubling time and its use as a measurement of prognosis and outcome. Furthermore, the use of common standards for prostate specific antigen doubling time in clinical trials is important as we determine which treatments should progress to randomized trials in which "hard" end points such as survival will be used. PMID- 18423745 TI - Prospective evaluation of the clinical usefulness of reflex fluorescence in situ hybridization assay in patients with atypical cytology for the detection of urothelial carcinoma of the bladder. AB - PURPOSE: The finding of atypical cytology is confusing for clinicians and it represents a management dilemma. We prospectively evaluated the clinical usefulness of fluorescence in situ hybridization assay for treating patients with atypical cytology who are at risk for bladder cancer. MATERIALS AND METHODS: Between May 2006 and June 2007 every patient who underwent cystoscopy and cytology with atypical or suspicious cytology underwent a reflex UroVysion test. A comprehensive review was then performed to evaluate clinical and pathological data on each patient. RESULTS: The population comprised 50 patients with no history of cancer and 70 who underwent cystoscopy for cancer surveillance. Fluorescence in situ hybridization assay was positive in all patients with cystoscopically visualized lesions whether they did or did not have a history of bladder cancer (positive predictive value 100%). In patients with equivocal cystoscopy and a history of cancer fluorescence in situ hybridization detected all 5 high grade tumors but it was false-negative for a low grade Ta tumor. In patients with equivocal cystoscopy and no prior cancer the positive predictive value was 50% and there was no false-negative assay result. In patients with negative cystoscopy and a history of cancer fluorescence in situ hybridization detected 3 cancers, including bladder carcinoma in situ in 2 and prostate carcinoma in situ in 1. In patients with no prior cancer and negative cystoscopy fluorescence in situ hybridization detected the only cancer (a high grade ureteral tumor) without yielding any false-negative results. CONCLUSIONS: This prospective evaluation of a reflex fluorescence in situ hybridization assay in patients with atypical cytology shows that the assay was unnecessary in patients with obvious tumors on cystoscopy but it was beneficial in patients with equivocal or negative cystoscopy. Fluorescence in situ hybridization assay identified all high grade cancers. It may help avoid unnecessary evaluation in patients with atypical cytology and equivocal or negative cystoscopy, while identifying those who would need further evaluation. PMID- 18423746 TI - Change in prostate specific antigen following androgen stimulation is an independent predictor of prostate cancer diagnosis. AB - PURPOSE: We tested the hypothesis that a single exogenous androgen injection in men with low prostate specific antigen would provoke a differential prostate specific antigen response that would correlate with the presence and volume of cancer at biopsy. MATERIALS AND METHODS: Following institutional review board approval 40 men with prostate specific antigen between 2.5 and 4.0 ng/ml were given 1 intramuscular injection of 400 mg testosterone cypionate at the start of the study. Prostate specific antigen and early morning serum testosterone were measured at baseline, 48 hours, and weeks 1, 2 and 4. All men underwent 12-core transrectal ultrasound guided biopsy at week 4. RESULTS: Of the 40 men 18 (45%) were diagnosed with prostate cancer. The mean change in prostate specific antigen from baseline to 4 weeks was 3.1 to 3.4 ng/ml (9.7%) in men found to have benign findings on biopsy compared to a mean increase of 2.9 to 3.8 ng/ml (29%) in those with prostate cancer (p = 0.006). The change in prostate specific antigen following androgen stimulation was significantly associated with the percent of tissue involved with cancer and it was an independent predictor of cancer diagnosis on univariate and multivariate analysis. CONCLUSIONS: An increase in prostate specific antigen following androgen stimulation in men with prostate specific antigen between 2.5 and 4.0 ng/ml was highly predictive of the subsequent diagnosis of prostate cancer and it correlated with disease volume. If these findings are corroborated, prostate specific antigen provocation may become an important strategy to identify men at risk for harboring prostate cancer and minimize the number undergoing unnecessary biopsies. PMID- 18423747 TI - Alfuzosin stone expulsion therapy for distal ureteral calculi: a double-blind, placebo controlled study. AB - PURPOSE: We evaluated the efficacy of alfuzosin as medical expulsive therapy for distal ureteral stone passage. MATERIALS AND METHODS: A total of 76 patients with a distal ureteral calculus provided consent for the study. Patients were randomized between placebo and study medication, and investigators and patients were blinded to the randomization scheme. Followup was done on a weekly basis and continued until the patient was rendered stone-free. The patient blood pressure, discomfort level, stone position on imaging, number of remaining pills and any adverse events were assessed. Statistical analysis was performed with the Student t test with p <0.05 considered significant. RESULTS: The overall spontaneous stone passage rate was 75%, including 77.1% for placebo and 73.5% for alfuzosin (p = 0.83). Mean +/- SD time needed to pass the stone was 8.54 +/- 6.99 days for placebo vs 5.19 +/- 4.82 days for alfuzosin. (p = 0.003). There was no difference in the size or volume of stones that passed spontaneously between the placebo and alfuzosin arms, as measured on baseline computerized tomography (4.08 +/- 1.17 and 3.83 +/- 0.95 mm, p = 0.46) and by a digital caliper after stone expulsion (3.86 +/- 1.76 and 3.91 +/- 1.06 mm, respectively, p = 0.57). When comparing the improvement from the baseline pain score, the alfuzosin arm experienced a greater decrease in pain score in the days after the initial emergency department visit to the date of stone passage (p = 0.0005). CONCLUSIONS: Alfuzosin improves the patient discomfort associated with stone passage and decreases the time to distal ureteral stone passage but it does not increase the rate of spontaneous stone passage. PMID- 18423748 TI - Phytotherapy for lower urinary tract symptoms secondary to benign prostatic hyperplasia. AB - PURPOSE: We examined the available data from clinical trials for certain botanicals used for lower urinary tract symptoms secondary to benign prostatic hyperplasia, including Serenoa repens (saw palmetto), Pygeum africanum (African plum), Secale cereale (rye pollen) and Hypoxis rooperi (South African star grass). MATERIALS AND METHODS: MEDLINE and The Cochrane Library searches were done in June 2007 using the terms benign prostatic hyperplasia, lower urinary tract symptoms, phytotherapy, saw palmetto, Serenoa, Permixon, Pygeum africanum, Tadenan, Cernilton, Cernitin and Hypoxis. Search results were assessed for relevance and the inclusion of placebo controlled trials. RESULTS: Two systematic reviews and 3 clinical trials were examined in the evaluation of Serenoa repens. Data from the systematic reviews showed an improvement in flow rates and symptoms. The results of 1 clinical trial were equivocal and the remaining 2 trials clearly showed equivalence to placebo. Systematic reviews were used in the evaluation of P. africanum, Secale cereale and Hypoxis rooperi. P. africanum and H. rooperi showed an improvement in flow rates and symptoms compared to placebo, while S. cereale showed an improvement in symptoms but not flow rates compared to placebo. CONCLUSIONS: Most clinical trials of investigating the efficacy of botanicals suffer from well documented methodological flaws. Saw palmetto has been clearly shown as comparable to placebo in a trial of sound methodology. While preliminary results appear promising, to our knowledge the remaining botanicals have yet to be evaluated in a trial of similar quality. PMID- 18423754 TI - Decreasing size at diagnosis of stage 1 renal cell carcinoma: analysis from the National Cancer Data Base, 1993 to 2004. AB - PURPOSE: The proportion of renal cell carcinoma cases diagnosed at stage I is known to be increasing significantly. We characterized stage I tumors further in terms of tumor size at diagnosis using a large national cancer registry. MATERIALS AND METHODS: The National Cancer Data Base captures approximately 75% of all newly diagnosed cancer cases in the United States. The database was queried for all adults who were diagnosed between 1993 and 2004 with stage I renal cell carcinoma. Trends were assessed in mean size with time as well as in the proportion of stage I tumors diagnosed at less than 2.0, less than 2.5 and less than 3.0 cm. RESULTS: There were 104,150 patients in the National Cancer Data Base diagnosed with stage I renal cell carcinoma during the study period. A total of 10,279 stage I tumors (9.9%) were less than 2.0 cm, 26,621 (25.6%) were 2.5 cm or less and 39,879 (38.3%) were 3.0 cm or less. Analysis of stage I renal cell carcinoma diagnoses with time demonstrated a statistically significant increase in the proportion of renal masses 3.0 cm or less between 1993 and 2004 (32.5% vs 43.4%). Of tumors 3.0 cm or less the proportion smaller than 2.0 cm increased significantly during the study period from 24.1% in 1993 to 29.4% in 2004. Mean tumor size decreased from 4.1 to 3.6 cm between 1993 and 2004 (p <0.001). CONCLUSIONS: Tumor size at diagnosis is decreasing with time in patients with stage I renal cell carcinoma. These data likely underestimate the proportion of all enhancing renal masses diagnosed at a small size. Patients with small masses may be appropriate candidates for nephron sparing surgery, energy based ablative therapy or active surveillance. Better technologies are needed to determine the diagnosis and prognosis of small enhancing renal masses. PMID- 18423755 TI - A new age in the management of vesicoureteral reflux? Infection rates after open surgery versus dextranomer injection. PMID- 18423756 TI - Pelvic floor dysfunction--what do women really want? PMID- 18423757 TI - The future of urological oncology: proceduralists or continued primary physicians and leaders. PMID- 18423758 TI - Positive surgical margins at partial nephrectomy: predictors and oncological outcomes. AB - PURPOSE: The prognostic significance and optimal management of positive surgical margins following partial nephrectomy remain ill-defined. We combine data from 2 tertiary care intuitions, and report predictors of positive surgical margins and long-term oncological outcomes for patients with positive surgical margins. MATERIALS AND METHODS: Clinical, pathological and followup data on 1,344 patients undergoing 1,390 partial nephrectomies for kidney cancer were analyzed. Patients with positive surgical margins on final pathology were treated expectantly. Univariate and multivariable logistic regression models were fit to determine clinicopathological features associated with positive surgical margins. The Kaplan-Meier method was used to estimate freedom from local disease recurrence and metastatic progression. Cox proportional hazards models were used to assess whether positive surgical margin predicted local recurrence or metastatic disease adjusting for tumor size, pathological stage, histological subtype and presence of a solitary kidney. RESULTS: Positive surgical margins were documented in 77 cases (5.5%). Decreasing tumor size and presence of a solitary kidney carried a significantly higher risk of positive surgical margins. The overall 10-year probability of freedom from local disease recurrence was 93% (95% CI 89, 95) and from metastatic progression 93% (95% CI 90, 95), with no significant difference between patients with positive vs negative margins (p = 0.97 and 0.18, respectively). Positive surgical margins were not associated with an increased risk of local recurrence or metastatic disease. CONCLUSIONS: Positive surgical margins in partial nephrectomy specimens do not uniformly portend an adverse prognosis. While every effort should be taken to ensure clear margins, our data suggest that select patients with a positive surgical margin can be safely offered vigilant monitoring without compromising long-term disease-free survival. PMID- 18423759 TI - Case-control study of medical comorbidities in women with interstitial cystitis. AB - PURPOSE: We used physician assigned diagnoses in an electronic medical record to assess comorbidities associated with interstitial cystitis. MATERIALS AND METHODS: A computer search of the administrative database at Kaiser Permanente Northwest, Portland, Oregon was performed for May 1, 1998 to April 30, 2003. All women with a medical record diagnosis of interstitial cystitis (ICD-9 code 595.1) were identified. These cases were then matched with 3 controls each based on age and duration in the health plan. The medical diagnoses (using ICD-9 codes restricted to 3 digits) assigned to these 2 groups were compared using the OR. RESULTS: A total of 239 cases and 717 matched controls were analyzed. There were 23 diagnoses that were significantly more common in cases than in controls (p < or = 0.005). Seven of these 23 diagnoses were other urological or gynecological codes used to describe pelvic symptoms. Additional specific conditions associated with interstitial cystitis were gastritis (OR 12.2), child abuse (OR 9.3), fibromyalgia (OR 3.0), anxiety disorder (OR 2.8), headache (OR 2.5), esophageal reflux (OR 2.2), unspecified back disorder (OR 2.2) and depression (OR 2.0). CONCLUSIONS: A diagnosis of interstitial cystitis was associated with multiple other unexplained physical symptoms and certain psychiatric conditions. Studies to explore the possible biological explanations for these associations are needed. Interstitial cystitis was also associated with a history of child abuse, although 96% of patients with IC did not have this diagnosis. PMID- 18423761 TI - A prospective study of lower urinary tract symptoms and erectile dysfunction. AB - PURPOSE: Several studies have shown that men with lower urinary tract symptoms are more likely to experience erectile dysfunction. All except 1 of these studies were cross-sectional, limiting inferences about whether lower urinary tract symptoms precipitate erectile dysfunction. MATERIALS AND METHODS: The association between lower urinary tract symptoms and incident erectile dysfunction was examined prospectively in the Health Professionals Follow-Up Study. Lower urinary tract symptoms were assessed biennially by the American Urological Association symptom index, which captures symptoms of frequency, urgency and force of urinary stream. Severe lower urinary tract symptoms was defined as a symptom score of 20 points or greater and no lower urinary tract symptoms was defined as a score of 7 points or less in men not treated for lower urinary tract symptoms. In 2000 the men were asked to rate erectile function for several periods. Erectile dysfunction was defined as poor or very poor function, or erectile dysfunction medication use, while no erectile dysfunction was defined as very good or good function and no erectile dysfunction medication use. We estimated the RR using Poisson regression, adjusting for age and other potentially confounding factors. RESULTS: We observed 3,953 incident erectile dysfunction cases among 17,086 men. Men with severe lower urinary tract symptoms in 1994 or earlier had a statistically significant 40% higher risk of erectile dysfunction subsequently than men without lower urinary tract symptoms. The risk of erectile dysfunction increased with increasing lower urinary tract symptom severity (p trend <0.0001). The positive association between lower urinary tract symptoms and erectile dysfunction was stronger in younger than in older men (p interaction = 0.03). CONCLUSIONS: This study provides evidence that men with lower urinary tract symptoms are more likely to have erectile dysfunction subsequently. PMID- 18423762 TI - Surgical versus nonsurgical treatment of women with pelvic floor dysfunction: patient centered goals at 1 year. AB - PURPOSE: In women with pelvic floor dysfunction we assessed the degree to which treatment (surgical vs nonsurgical) was associated with achievement of patient centered goals, satisfaction with care and quality of life. MATERIALS AND METHODS: In this prospective cohort study between September 2003 and December 2004 we recruited women during their first referral visit for pelvic floor dysfunction treatment at our outpatient Urogynecology Clinic. At the first visit women enumerated up to 5 personal treatment goals, and anchored each goal by anticipating best and worst possible outcomes. At 12-month followup women were asked to indicate the level of goal attainment (-2 worst outcome, +2 best outcome). At baseline and followup women completed short forms of the Incontinence Impact Questionnaire and Urogenital Distress Inventory (range 0 to 100, high scores indicating greater impact or distress). Patients indicated the level of treatment satisfaction on a 4-level ordinal scale. RESULTS: Of the 127 study participants with complete data 46 (36.2%) were treated surgically and 81 (63.8%) were treated nonsurgically. There were no major demographic differences between the 2 groups in terms of age, race, weight, prior pelvic floor dysfunction surgery and vaginal parity. The surgical group was more likely to have received a baseline diagnosis of pelvic organ prolapse (80% vs 60%, p = 0.0259) and be postmenopausal (89% vs 72%, p = 0.0261). There were no significant differences in the distribution of goal type (symptom relief, activity, self image, general health) by treatment status (p = 0.1074). Using logistic regression to adjust for age and baseline diagnosis, surgically treated patients at 1 year were significantly more likely to report complete primary goal attainment (OR 4.42, p = 0.0154) and complete treatment satisfaction (OR 6.12, p = 0.0109). For all participants 1-year Incontinence Impact Questionnaire-7 and Urogenital Distress Inventory-6 scores were significantly correlated with primary goal attainment scores. CONCLUSIONS: In this nonrandomized, prospective analysis surgically treated patients with pelvic floor dysfunction had higher 1-year self described complete goal attainment and satisfaction scores compared with patients treated nonsurgically. Goal attainment scores correlated with disease specific quality of life. Patient centered outcomes should be incorporated in multicenter prospective research trials for pelvic floor disorders and in clinical practice to inform treatment plans. PMID- 18423763 TI - Evoked cavernous activity: normal values. AB - PURPOSE: We present normative data for evoked cavernous activity, an electrodiagnostic test that evaluates the autonomic innervation of the corpora cavernosa. MATERIALS AND METHODS: We enrolled 37 healthy, sexually active and potent men for the study. Each subject completed an International Index of Erectile Function questionnaire, and underwent simultaneous evoked cavernous activity and hand and foot sympathetic skin response testing. The sympathetic skin response tests were performed as autonomic controls. RESULTS: A total of 36 men had discernible evoked cavernous activity and sympathetic skin responses. The mean International Index of Erectile Function erectile domain score was 27. Evoked cavernous activity is a low frequency wave that is morphologically and temporally similar in both corpora. The amplitudes of the responses were highly variable. The latencies, although variable, always occurred after the hand sympathetic skin response. There was no change in the quality or the latency of the evoked cavernous activity with age. CONCLUSIONS: Evoked cavernous activity is measurable in healthy, potent men in a wide range of ages. Similar to other evoked responses of the autonomic nervous system, the measured waveform is highly variable but its presence is consistent. The association between evoked cavernous activity and erectile function is to be determined. PMID- 18423764 TI - Comparing the antimicrobial effectiveness of NaCl and KCl with a view to salt/sodium replacement. AB - A study using a small range of pathogenic bacterial species (Aeromonas hydrophila, Enterobacter sakazakii, Shigella flexneri, Yersinia enterocolitica and 3 strains of Staphylococcus aureus) has shown that potassium chloride has an equivalent antimicrobial effect on these organisms when calculated on a molar basis. Combined NaCl and KCl experiments were carried out and data was analysed using a modification to the Lambert and Lambert [Lambert, R.J.W., and Lambert, R., 2003. A model for the efficacy of combined inhibitors. Journal of Applied Microbiology 95, 734-743.] model for combined inhibitors and showed that in combination KCl is a direct 1:1 molar replacement for the antimicrobial effect of common salt. If this is a general finding then, where salt is used to help preserve a product, partial or complete replacement by KCl is possible. PMID- 18423765 TI - Normal sublingual microcirculation during painful crisis in sickle cell disease. AB - Obstruction of the microcirculation is the most important cause of painful crisis in sickle cell disease (SCD). Extensive microvascular obstruction has been observed in mouse models of SCD. A technique to determine the extent of the microcirculatory obstructions in humans may be helpful in the clinical setting and for research purposes. Therefore, we measured sublingual microcirculation longitudinally in patients with SCD admitted with painful crisis. Sublingual microcirculation was recorded with side-stream darkfield (SDF) imaging and semi quantified with a microvascular flow index (MFI) on a range from 0 to 4 (arbitrary units; from 0 (no flow) to 4 (hyperdynamic flow)). Thirteen consecutive adult sickle cell patients admitted with painful crises were included and provided 47 measurements of MFI in 14 episodes of painful crisis. Seven patients provided baseline measurements and seven healthy controls were studied. The mean (+/-standard error of the mean) MFI during painful crisis was 2.6+/-0.1 and did not change during the painful crisis. The mean MFI of patients with SCD during steady state (2.7+/-0.1) and the mean MFI of the controls (2.7+/-0.1) were not different from the mean MFI during painful crisis. During painful crisis irregular microvascular perfusion, expressed by the distribution width of the microvascular blood flow velocity, correlated negatively (r=-0.484; P=0.002) with hemoglobin concentration. We conclude that sublingual microcirculatory blood flow velocity is not disturbed in sickle cell patients during painful crisis. PMID- 18423766 TI - Increase of the pharmacological and pharmacokinetic efficacy of negatively charged polypeptide recombinant hirudin in rats via parenteral route by association with cationic liposomes. AB - Two biodegradable cationic lipids, stearylamine and DC-Chol, were chosen to investigate the effect of cationic lipids on the in vitro and in vivo characteristics of hydrophilic proteins or peptides of low isoelectric point. Thrombin inhibitor recombinant hirudin variant-2 (rHV2) was selected as the model drug. The cationic lipids were found to achieve higher entrapment efficiency of rHV2 in liposomes than zwitterionic lipids. The positively charged liposomes became less positive and relatively stable in serum after loading rHV2. The cationic liposomes induced sustained release of rHV2 in the presence of plasma, significantly prolonged the antithrombotic efficacy and plasma level of rHV2 after intravenous injection in rats in comparison with neutral lipid liposomes, especially for stearylamine group. Both clotting times correlated well with plasma rHV2 levels. No serious adverse events were observed and physical state of rats was satisfactory for all the formulations. Electrostatic interaction between negative charge of rHV2 and cationic liposomes was confirmed and it might affect all the characteristics of rHV2 loaded cationic vehicles. The findings suggest that cationic liposomes may be a potential sustained-release delivery system for parenteral administration of hydrophilic proteins or peptides with low isoelectric point to prolong efficacy and improve bioavailability. PMID- 18423767 TI - A novel bottom-up process to produce drug nanocrystals: controlled crystallization during freeze-drying. AB - To improve the dissolution behavior of lipophilic drugs, a novel bottom-up process based upon freeze drying which allows for the production of nanocrystalline particles was developed: "controlled crystallization during freeze drying". This novel process could strongly increase the dissolution behavior of fenofibrate. For example at a drug load of 30% w/w, 80% of the drug dissolved within 10 min from tablets prepared from the controlled crystallized dispersions, while from tablets prepared from the physical mixture only 50% was dissolved after 120 min. Furthermore it was found that faster freezing or using a solution with a lower water/tertiary butyl alcohol (TBA) ratio resulted in faster dissolution, indicating that the crystalline dispersions contained smaller crystals. Crystallization of the drug could occur during freezing or during drying. When crystallization occurs during freezing, faster freezing or using solutions with a lower water/TBA ratio results in the formation of more nuclei and consequently smaller crystals. When crystallization occurs during drying, faster freezing or using solutions with a higher water/TBA ratio results in the formation of smaller solvent crystals and therefore smaller interstitial spaces which contain the freeze-concentrated fraction. Since crystallization occurs in the freeze-concentrated fraction and the size of the crystals are limited to the size of the interstitial spaces, smaller crystals are formed in these situations. PMID- 18423768 TI - Influence of oil content on physicochemical properties and skin distribution of Nile red-loaded NLC. AB - The aims of this study were to investigate the effect of the oil content on the physicochemical properties of NLC and to elucidate the potential of NLC for skin targeting. The obtained results showed that an increase in the oil content did not affect the mean particle size of NLC but impacted on the zeta potential. The inner structure of NLC was influenced by the increasing proportion of oil towards the less ordered structure as confirmed by differential scanning calorimetry (DSC) and X-ray diffraction (XRD), particularly for the higher medium chain triglycerides (MCT) loading. The data from proton nuclear magnetic resonance (1H NMR) revealed that cetyl palmitate nanoparticles did not completely recrystallize after cooling down to room temperature. 1H NMR and DSC results indicate that MCT molecules were restricted in the NLC as compared to the nanoemulsions (NE). Nile red distribution and penetration into skin from NLC were pronounced as compared to NE and dependent on the MCT loading. The deep penetration and high amount of Nile red were related to the occlusion factor. Moreover, the epidermal targeting was achieved by NLC applications, particularly those containing 5% MCT (NLC-5) depending on the amount of MCT loading. PMID- 18423770 TI - Stage-specificity of spontaneous mutation at a tandem repeat DNA locus in the mouse germline. AB - Mouse expanded simple tandem repeat (ESTR) loci are the most unstable loci in the mouse genome. Despite the fact that over the last decade these loci have been extensively used for studying germline mutation induction in mice, to date little is known about the mechanisms underlying spontaneous and induced ESTR mutation. Here we used flow cytometry and single-molecule PCR to compare the frequency of ESTR mutation in four flow-sorted fractions of the mouse male germ cells - spermatogonia, spermatocytes I, round and elongated spermatids. The frequency and the spectrum of ESTR mutation did not significantly differ between different stages of mouse spermatogenesis. Considering these data and the results of other publications, we propose that spontaneous ESTR mutation is mostly attributed to replication slippage in spermatogonia and these loci may be regarded as a class of expanded microsatellites. PMID- 18423769 TI - Diesel exhaust particles are mutagenic in FE1-MutaMouse lung epithelial cells. AB - The particulate phase of diesel engine exhaust is likely carcinogenic. However, the mechanisms of diesel exhaust particles (DEPs) induced mutagenicity/carcinogenicity are still largely unknown. We determined the mutant frequency following eight repeated 72 h incubations with 37.5 or 75 microg/ml DEP (NIST SRM 1650) in the FE1-MutaMouse lung epithelial cell line. We measured DEP induced acellular and intracellular production of reactive oxygen species (ROS) and compared with ROS production induced by carbon black, which we have previously shown is mutagenic in this cell line [N.R. Jacobsen, A.T. Saber, P. White, P. Moller, G. Pojana, U. Vogel, S. Loft, J. Gingerich, L. Soper, G.R. Douglas, H. Wallin. Increased mutant frequency by carbon black, but not quartz, in the lacZ and cII transgenes of mutamouse lung epithelial cells, Environ. Mol. Mutagen. 48(6) (2007) 451-461]. The mutant frequency was marginally elevated in cells treated with 37.5 microg/ml DEP (1.29-fold [95% CI: 0.96-1.60], p=0.08) and significantly increased in cells treated with 75 microg/ml DEP (1.55-fold [95% CI: 1.23-1.87], p < 0.001). ROS production from DEP was low both within cells and in acellular systems when compared to carbon black. These results show that DEP are mutagenic in a mammalian cell line in vitro and that additional pathways besides ROS production, such as those involving the presence of polycyclic aromatic hydrocarbons, likely are involved in the mutagenesis. PMID- 18423771 TI - Diagnostic strategy to improve the efficiency of screening diabetes--ROC analysis for fasting plasma glucose. PMID- 18423772 TI - [Electrophysiological investigations in childhood epilepsy surgery]. AB - Epilepsy surgery in children is a functional surgery: its goal is to perform the resection of the epileptic brain tissue while sparing the eloquent cortex. Prolonged scalp video-EEGs allow recording of all types of seizures and play a crucial role in localizing the epileptogenic zone. Furthermore, EEG data correlation with clinical and radiological findings provides a guide for the surgical strategy: either resection without further investigations or an invasive recording procedure. In prehemispherotomy evaluation, EEG recordings confirm that limited resections are not indicated and demonstrate that the opposite hemisphere is not involved. If invasive recordings are needed, they consist in foramen ovale electrode insertion, which provides valuable information in mesial temporal lobe epilepsy, stereoelectroencephalography for children older than two years, and subdural grids associated with depth electrodes in infants or when the eloquent areas need to be carefully investigated. Such investigations allow tailoring surgery to each child. PMID- 18423773 TI - [Vagus nerve stimulation for the treatment of refractory epilepsy]. AB - Proposed as an additive symptomatic treatment of refractory epilepsy, vagus nerve stimulation (VNS) has proven to be effective and well-tolerated in patients presenting with refractory epilepsy for whom cortical resection is not indicated. After two years of treatment, the overall reduction of seizure frequency averaged 40%. In 50% of the patients, the frequency of seizures decreased by at least 50%. Moreover, even in absence of a significant reduction of seizures, patients who undergo this treatment reported an improvement in their quality of life. Economic surveys also demonstrate a favorable impact of VNS on the management of refractory epilepsy. Since 1988, 65,000 patients with refractory epilepsy throughout the world have been treated by VNS for this indication (1000 in France). The surgical implantation technique used in our department, the effects of vagus nerve stimulation reported in the literature, and our experience with a cohort of 70 patients with refractory epilepsy who received implants over the last 10 years are described. PMID- 18423774 TI - [Long-term outcome after hemispheric disconnection]. AB - Hemispheric disconnection has been largely proposed for patients with severe epilepsy associated with a congenital or acquired hemispheric cerebral pathology. The classical procedure of anatomical hemispherectomy was progressively abandoned by neurosurgeons in order to avoid postoperative complications since then hemispherotomy techniques have been developed. Globally, with hemispheric disconnection, the rate of patients becoming seizure-free has been between 50 and 80%. The factors affecting seizure control have not been completely elucidated, but several authors suggested that differences in etiology as well as the hemispheric disconnection technique used may partially explain this variability. The percentage of seizure-free patients is higher with hemispherotomy techniques and in the group of patients with Rasmussen encephalitis, Sturge-Weber syndrome, and vascular insults. Depending on overall long-term progression, there is an improvement compared to preoperative status even if children exhibit heterogenous abilities. The lowest scores are observed for motor skills but communication and socialization are relatively well-preserved and strongly related to the duration of epilepsy: the longer the duration, the lower the scores were. Neuropsychological outcome following hemispheric disconnection makes it possible to study the development of hemispheric specialization during infancy and to provide information on cognitive recovery. Cerebral reorganization has been proved to exist in motor and language recovery. Ipsilateral corticospinal pathways seem to be involved in the movement of hemiplegic limbs. Everyday language can be supported by both hemispheres, but there is an early hemispheric specialization of the left hemisphere according to metaphonologic abilities. PMID- 18423775 TI - Changes in out-of-pocket payments for healthcare in Vietnam and its impact on equity in payments, 1992-2002. AB - BACKGROUND: Economic reforms in Vietnam initiated in the late 1980s included deregulation of the health system resulting in extensive changes in health care delivery, access, and financing. One aspect of the health sector reform was the introduction of user fees at both public and private health facilities, which was in stark contrast to the former socialized system of free medical care. Subsequently, health insurance and free health care cards for the poor were introduced to mitigate the barriers to seeking care and financial burden imposed by out-of-pocket (OOP) health payments as a result of the user fees. OBJECTIVE: To examine the determinants of seeking care and OOP payments as well as the relationship between individual out-of-pocket (OOP) health expenditures and household ability to pay (ATP) during 1992-2002. DATA: The data are drawn from 1992-93 and 1997-98 Vietnam Living Standard Surveys (VLSS) and 2002 Vietnam Household and Living Standards Survey (VHLSS). METHODS: We use a two-part model where the first part is a probit model that estimates the probability that an individual will seek treatment. The second part is a truncated non-linear regression model that uses ordinary least-squares and fixed effects methods to estimate the determinants of OOP payments that are measured both as absolute as well as relative expenditures. Based on the analysis, we examine the relationship between the predicted shares of individual OOP health payments and household's ATP as well as selected socioeconomic characteristics. RESULTS: Our results indicate that payments increased with increasing ATP, but the consequent financial burden (payment share) decreased with increasing ATP, indicating a regressive system during the first two periods. However, share of payments increased with ATP, indicating a progressive system by 2002. When comparing across years, we find horizontal inequities in all the years that worsened between 1992 and 1998 but improved by 2002. CONCLUSION: The regressivity in payments noted during 1992 and 1998 might be because the rich could avail of health insurance more than those at lower incomes and as a consequence, were able to use the healthcare system more effectively without paying a high OOP payment. In contrast, the poor either incurred higher OOP payments or were discouraged from seeking treatments until their ailment became serious. This inequality becomes exacerbated in 1998 when insurance take-up rates were not high, but the impact of privatization and deregulation was already occurring. By 2002, insurance take-up rates were much higher, and poverty alleviation policies (e.g., free health insurance and health fund membership targeted for the poor) were instituted, which may have resulted in a less regressive system. PMID- 18423776 TI - Neurokinin B/NK3 receptors exert feedback inhibition on L-DOPA actions in the 6 OHDA lesion rat model of Parkinson's disease. AB - Neurokinin B (NKB) and substance P (SP) act via NK(3) and NK(1) receptors. Using the unilateral 6-hydroxydopamine (6-OHDA) lesion rat model of Parkinson's disease (PD), it was found that chronic, but not acute, administration of L-DOPA increases striatal NKB expression in the dopamine-depleted hemisphere. In contrast, both acute and chronic administrations of L-DOPA restore reduced levels of SP mRNA. Co-treatment with the NK(3) receptor antagonist, SB222200, and L-DOPA increased contralateral rotations compared to L-DOPA alone in L-DOPA primed rats. The NK(3)R agonist, senktide, increased the phosphorylation of tyrosine hydroxylase (TH) at Ser(19)-TH, a CaMKII site, and of Thr(286)-CaMKII in striatal slices. Senktide had no effect on P-Ser(31)-TH, a MAPK site, but reduced P Ser(217/221)-MEK. Amperometry demonstrated that senktide increased evoked dopamine release. SB222200 blocked the effects of senktide. In striatal slices prepared from 6-OHDA-lesioned rats repeatedly treated with L-DOPA, senktide no longer increased P-Thr(286)-CaMKII, suggesting a role of NK(3)R on dopamine terminals under normal conditions. SB222200 increased P-Ser(217/221)-MEK only in dopamine-depleted slices, indicating an increased NK(3)R tone under Parkinsonism conditions. Altogether, these data demonstrate a differential regulation of NKB and SP by L-DOPA in an animal model of PD and indicate a unique role of NKB in long-term effects of L-DOPA. Behavioural, biochemical and amperometric data indicate that NKB/NK(3)R signalling stimulates dopamine transmission at the presynaptic site, but inhibits it at the postsynaptic site. The inhibitory influence of NKB/NK(3)R on dopamine transmission dominates in an animal model of PD and provides a feedback inhibition on actions mediated via L-DOPA. PMID- 18423777 TI - WAY-200070, a selective agonist of estrogen receptor beta as a potential novel anxiolytic/antidepressant agent. AB - Recent studies have reported that estrogen has antidepressant-like effects in animal models. In this study we used the highly selective ER beta agonist, WAY 200070, to examine the role of ER beta activation on brain neurochemistry and activity in antidepressant and anxiolytic models in male mice. Within 15 min of administration, WAY-200070 (30 mg/kg s.c.) caused the nuclear translocation of striatal ER beta receptors from the cytosol. WAY-200070 also increased c-fos activation 4h, but not 15 min after administration. Both nuclear translocation and c-fos induction effects of WAY-200070 demonstrate that WAY-200070 has bound to estrogen receptors and triggered downstream events. The absence of these effects in the ER beta KO mice confirms that WAY-200070 was targeting ER beta. Administration of WAY-200070 (30 mg/kg s.c.) produced a delayed approximately 50% increase in dopamine in the striatum of wild type mice. The effect was significant and maintained from 90 to 240 min. This increase was absent in ER beta KO mice. In wild type mice, WAY-200070 (30 mg/kg s.c.) also produced a delayed and transient approximately 100% increase in 5-HT. To further investigate the role of ER beta receptors on serotonergic function, 5-HTP accumulation was measured. ER beta KO mice were found to have reduced frontal cortex levels of 5 HTP, indicating reduced tryptophan hydroxylase activity. WAY-200070 (3-30 mg/kg s.c.) was also tested in behavioural models. WAY-200070 (30 mg/kg s.c.) reduced immobility time in the mouse tail suspension test indicating an antidepressant like effect. WAY-200070 (30 mg/kg) showed anxiolytic-like effects in the four plate test (increased punished crossings) and stress-induced hyperthermia (attenuation of hyperthermic response). The effects of the selective ER beta agonist, WAY-200070, on dopamine and serotonin, the anxiolytic-like and antidepressant-like effects as well as the genotype specific effects on neurochemistry support that positive modulation of ER beta function may provide a novel treatment for affective disorders. PMID- 18423778 TI - An in vivo [18F]MK-9470 microPET study of type 1 cannabinoid receptor binding in Wistar rats after chronic administration of valproate and levetiracetam. AB - There is substantial evidence that the endocannabinoid system and in particular the type 1 cannabinoid receptor (CB1R) is involved in epilepsy. We evaluated the in vivo effect of chronic administration of the anti-epileptic drugs valproate (VPA) and levetiracetam (LEV) on rat brain CB1 receptors using the positron emission tomography (PET) tracer [(18)F]MK-9470. Six Wistar rats were treated with VPA (200mg/kg) or LEV (50mg/kg) IP daily for 2 weeks. Dynamic imaging after intravenous injection of 18 MBq [(18)F]MK-9470 was performed on a FOCUS 220 microPET at baseline and after chronic treatment. Six animals were used as controls and were injected with saline, using the same protocol. Parametric images based on standardized uptake values (SUV) were generated and were spatially normalized to Paxinos space. These CB1R images were analyzed using a predefined volume of interest (VOI)-based analysis. Differences in SUV values between chronic and baseline scans in each condition (saline, VPA and LEV treatment) were calculated in each VOI. Direct binding affinity of the drugs at CB1R was assessed by competitive binding assay in Chinese hamster ovarian cells expressing human CB1R. Chronic injections of saline did not produce significant changes in global [(18)F]MK-9470 binding (p=0.43), nor in tracer binding in individual VOIs. We found a significant increase in global cerebral [(18)F]MK 9470 binding after chronic VPA administration compared to sham treated animals (+32.5%, p<0.001), as well as in tracer binding in all individual VOIs. After chronic administration of LEV, there was no significant change in global cerebral CB1R binding (+6.9%, p=0.81), nor in tracer binding in individual VOIs. As VPA does not exhibit high affinity for CB1R (displacement of [(3)H]-SR141716A 1.3+/ 14.0%), such upregulation is most likely caused by an indirect effect on the endocannabinoid system. This increase in CB1R tracer binding and possibly signaling may represent a supplementary and new mechanism of VPA, but not LEV, since activation of CB1Rs has been shown to decrease excitability and excitotoxicity on-demand. PMID- 18423779 TI - Protein nanoparticles as drug carriers in clinical medicine. AB - Solvent-based delivery vehicles for chemotherapy agents have been instrumental in providing a means for hydrophobic agents to be administered intravenously. These solvents, however, have been associated with serious and dose-limiting toxicities. Solvent-based formulations of taxanes, a highly active class of cytotoxic agents, are associated with hypersensitivity reactions, neutropenia, and neuropathy. Nanoparticle technology utilizing the human protein albumin exploits natural pathways to selectively deliver larger amounts of drug to tumors while avoiding some of the toxicities of solvent-based formulations. 130 nM albumin-bound (nab) paclitaxel (nab-paclitaxel; Abraxane) was recently approved for use in patients with metastatic breast cancer who have failed combination therapy. In a randomized, phase III study in metastatic breast cancer, nab paclitaxel was found to have improved efficacy and safety compared with conventional, solvent-based paclitaxel. Preliminary data also suggest roles for nab-paclitaxel as a single agent and in combination therapy for first-line treatment of metastatic breast cancer as well as in other solid tumors, including non-small-cell lung cancer, ovarian cancer, and malignant melanoma. The nab technology promises to have broad utility in cancer therapy, and clinical trials are underway using nab formulations of other water-insoluble anticancer agents such as docetaxel and rapamycin. PMID- 18423780 TI - Prion protein M129V polymorphism affects retrieval-related brain activity. AB - The prion protein Met129Val polymorphism has recently been related to human long term memory with carriers of either the 129MM or the 129MV genotype recalling 17% more words than 129(VV) carriers at 24h following learning. Here, we sampled genotype differences in retrieval-related brain activity at 30min and 24h following learning. Furthermore, genotype groups were compared regarding grey matter concentrations and cognitive profiles. We used event-related functional magnetic resonance imaging (fMRI) during a word recognition task on 12 Met/Met carriers, 12 Val/Met carriers, and 12 Val/Val carriers. These groups were matched for retrieval performance, gender, age, education, and other memory-related genetic polymorphisms. Although retrieval performance was matched, Val carriers exhibited enhanced retrieval-related brain activity at 30min and 24h following learning. At both time lags, correlations between retrieval-related brain activity and retrieval success were negative for Val homozygotes (the more activity, the worse retrieval success), while correlations showed no significance or were positive for Met homozygotes and heterozygotes. These results suggest a less economic use of retrieval-related neural resources in Val relative to Met carriers. Furthermore, Val carriers exhibited higher neocortical grey matter concentrations compared to Met carriers. When controlling for grey matter concentration, genotype effects in retrieval-related brain activity remained significant. Val and Met carriers yielded comparable brain activations for correct rejections of non-studied words and for working memory, which speaks to the specificity of the genotype effect. Findings suggest that the prion protein Met129Val polymorphism affects neural plasticity following learning at a time scale of minutes to hours. PMID- 18423781 TI - High efficacy of erlotinib in Taiwanese NSCLC patients in an expanded access program study previously treated with chemotherapy. AB - PURPOSE: Erlotinib is the first epidermal growth factor receptor-tyrosine kinase inhibitor (EGFR-TKI) which has demonstrated a survival benefit in non-small-cell lung cancer (NSCLC) patients. An open label phase II study was conducted in Taiwanese patients with NSCLC to evaluate its efficacy. METHODS: Patients with proven stage IIIB/IV NSCLC who had received at least one line of standard chemotherapy or radiotherapy were enrolled into this study. All patients were given oral erlotinib, 150mg/day till disease progression. RESULTS: From May 2005 to July 2006, 300 patients were entered from 14 hospitals in Taiwan. This analysis was based on 299 patients who received at least one dose of erlotinib. The best response rates were a 29% partial response and 44% stable disease in 273 patients who had response data available. Non-smoking (p=0.033), adenocarcinoma/BAC (p=0.0027), female (p=0.0013), aged less than 65 years (p=0.0115), stage IV (p=0.0492), patients with skin rash (p=0.0216), and a higher grade of skin rash (p=0.003) were significantly correlated with response to treatment. Skin rash was a common adverse event (any grade: 84%, Gr 3-4: 16%). The median time to disease progression was 5.6 months. Cox regression model for progression free survival showed patients most at risk of early progression were males of low performance status having squamous cell carcinoma. CONCLUSIONS: This was the largest multicenter prospective clinical study of NSCLC in Taiwan. The results demonstrated the excellent response rates, time-to-progression and overall survival of erlotinib in a large population of Taiwanese NSCLC patients who had been previously treated with chemotherapy or radiotherapy. PMID- 18423782 TI - The safety and efficacy of gefitinib versus platinum-based doublets chemotherapy as the first-line treatment for advanced non-small-cell lung cancer patients in East Asia: a meta-analysis. AB - BACKGROUND: To evaluate the risk/benefit profiles of gefitinib in comparison with platinum-based doublets chemotherapy as a first-line treatment for chemonaive patients with advanced non-small-cell lung cancer in East Asia. METHODS: We searched MEDLINE, EMBASE, Cochrane Library, and ClinicalTrials.gov to identify randomized and non-randomized phase II or III clinical trials of gefitinib or chemotherapy treatment in East Asian patients published before 4/30/2007. Two reviewers independently applied selection criteria, performed quality assessment, and extracted data. Treatment arms with gefitinib 250mg/day and platinum-based doublets chemotherapy irrespective of dosage and schedule were combined to calculate the pooled estimates for efficacy and safety outcomes of interest. RESULTS: We identified 7 gefitinib and 41 platinum-based doublets chemotherapy trials with nearly 3000 enrolled patients for planned comparison. The pooled response rate (95% confidence interval) to gefitinib for unselected chemonaive population was 31% (23-38%), not substantially different from 34% (31-38%) reported by platinum-based doublets chemotherapy trials. Patients with certain characteristics were more likely to benefit from gefitinib treatment, with pooled response rates as high as 75% (60-90%) for patients with epidermal growth factor receptor (EGFR) exon 18-21 mutations; 56% (38-74%) for never smokers; 55% (41 69%) for female; and 43% (30-57%) for adenocarcinoma or bronchioalveolar carcinoma. Severe hematological adverse events related to gefitinib treatment were not observed in any of the included trials. However, the risks of severe liver and lung injury related to gefitinib treatment were both approximately 6%, significantly higher than 1% and 0.2% reported by platinum-based doublets chemotherapy trials. CONCLUSION: Our data suggest that one third of chemonaive NSCLC patients in East Asia would respond to oral gefitinib monotherapy while 6% would develop severe liver and lung injury. Although patients with EGFR gene mutations, female gender, non-smokers, or adenocarcinoma were more likely to respond to gefitinib, further study with valid comparison groups are needed to identify the optimal treatment strategy in these subpopulations. PMID- 18423783 TI - Change of color and components in sepals of chameleon hydrangea during maturation and senescence. AB - The sepal color of a chameleon hydrangea, Hydrangea macrophylla cv. Hovariatrade mark 'Homigo' changes in four stages, from colorless to blue, then to green, and finally to red, during maturation and the senescence periods. To clarify the chemical mechanism of the color change, we analyzed the components of the sepals at each stage. Blue-colored sepals contained 3-O-sambubiosyl- and 3-O glucosyldelphinidin along with three co-pigments, 5-O-p-coumaroyl-, 5-O-caffeoyl- and 3-O-caffeoylquinic acids. The contents of glycosyldelphinidins decreased toward the green-colored stage, with a coincident increase in the number of chloroplasts. During the last red colored stage, the two species of 3-O glycosyldelphinidin almost disappeared, and another two anthocyanins, 3-O sambubiosyl- and 3-O-glucosylcyanidin, increased in amounts. Mixing of 3-O glycosylcyanidins, co-pigments, and Al3+ in a buffered solution at pH 3.0-3.5 gave not a blue, but a red, colored solution that was the same as that of the sepal color of the 4th stage. Sepals of hydrangea grown in an highland area also turned red in autumn, and contained the same cyanidin glycosides. The red coloration of the hydrangea during senescence was due to a change in anthocyanin biosynthesis. PMID- 18423784 TI - Analytical solutions for flow fields near continuous wall reactive barriers. AB - Permeable reactive barriers (PRBs) are widely applied for in-situ remediation of contaminant plumes transported by groundwater. Besides the goal of a sufficient contaminant remediation inside the reactive cell (residence time) the width of plume intercepted by a PRB is of critical concern. A 2-dimensional analytical approach is applied to determine the flow fields towards rectangular PRBs of the continuous wall (CW) configuration with and without impermeable side walls (but yet no funnel). The approach is based on the conformal mapping technique and assumes a homogeneous aquifer with a uniform ambient flow field. The hydraulic conductivity of the reactive material is furthermore assumed to exceed the conductivity of the aquifer by at least one order of magnitude as to neglect the hydraulic gradient across the reactor. The flow fields are analyzed regarding the widths and shapes of the respective capture zones as functions of the dimensions (aspect ratio) of the reactive cell and the ambient groundwater flow direction. Presented are an improved characterization of the advantages of impermeable side walls, a convenient approach to improved hydraulic design (including basic cost optimization) and new concepts for monitoring CW PRBs. Water level data from a CW PRB at the Seneca Army Depot site, NY, are used for field demonstration. PMID- 18423785 TI - Nutrient dynamics as indicators of karst processes: comparison of the Chalk aquifer (Normandy, France) and the Edwards aquifer (Texas, U.S.A.). AB - Karst aquifers display a range of geologic and geomorphic characteristics in a wide range of climatic and land-use settings; identification of transport dynamics representative of karst aquifers in general could help advance our understanding of these complex systems. To this end, nutrient, turbidity, and major ion dynamics in response to storms were compared at multiple sites in two karst aquifers with contrasting characteristics and settings: the Chalk aquifer (Eure Department, Normandy, France) and the Barton Springs segment of the Edwards Aquifer (Texas, U.S.A.). The Chalk aquifer is typified by high matrix porosity, thick surficial deposits (up to 30 m thick), and agricultural land use; the Barton Springs segment is typified by low matrix porosity, outcropping limestone, and urban land use. Following one to three storms, from 5 to 16 samples from springs and wells were analyzed for major ions, and specific conductance and turbidity were monitored continuously. Comparison of the chemographs indicated some generalized responses, including an increase in turbidity and potassium concentrations and a decrease in major ion and nitrate concentrations with infiltrating storm runoff. Factor analysis of major ions and turbidity revealed strikingly similar behavior of the chemical variables for the two aquifers: The first two factors, explaining more than 75% of the variability, illustrate that dynamics of most major ions (including nitrate) are opposed to those of turbidity and of potassium. The results demonstrate that potassium and nitrate are effective tracers of infiltrating storm runoff and resident ground water, respectively, and the similar results for these two highly contrasting aquifers suggest that the dynamics identified might be applicable to karst systems in general. PMID- 18423786 TI - Gas chromatography-mass spectrometry method for the analysis of 19-nor-4 androstenediol and metabolites in human plasma: application to pharmacokinetic studies after oral administration of a prohormone supplement. AB - 19-Nor-4-androstenediol is a prohormone of nandrolone. Both substances are included in the WADA list of prohibited classes of substances. The aim of this study is to determine the plasma levels of 19-nor-4-androstenediol and its metabolites after oral administration of a nutritional supplement containing the drug. Two capsules of Norandrodiol Select 300 were orally administered to six healthy male volunteers. Plasma samples were collected up to 24h. Samples were extracted to obtain free and glucuronoconjugated metabolic fractions. Trimethylsilyl derivatives of both fractions were analyzed by gas chromatography coupled to mass spectrometry (GC-MS). The method was validated to determine linearity, extraction recovery, limit of detection and quantification, intra- and inter-day precision and accuracy. After administration of 19-nor-4 androstenediol, the main metabolites detected were norandrosterone and noretiocholanolone, mainly in the glucuronide fraction. Nandrolone, norandrostenedione and 19-nor-4-androstenediol were also detected at lower concentrations. PMID- 18423787 TI - Down-regulation of flavonoid 3'-hydroxylase gene expression by virus-induced gene silencing in soybean reveals the presence of a threshold mRNA level associated with pigmentation in pubescence. AB - Changes in flavonoid content are often manifested as altered pigmentation in plant tissues. Two loci have been identified as controlling pigmentation in soybean pubescence. Of these, the T locus appears to encode flavonoid 3' hydroxylase (F3'H) protein: the T and t alleles are associated with tawny and gray colors, respectively, in pubescence. We previously down-regulated F3'H gene expression by virus-induced gene silencing (VIGS) in soybean. Despite this successful VIGS, the tawny pubescence pigmentation proved to be unchanged in greenhouse-grown plants. We hypothesized that the reduced mRNA level of the F3'H gene resulting from VIGS remained high enough to induce pigmentation. To verify this hypothesis, in the present study, we performed F3'H VIGS on plants grown under controlled conditions, in which the steady-state mRNA level of the F3'H gene was reduced to approximately 5% of that of greenhouse-grown plants. This VIGS treatment resulted in the loss of tawny pigmentation in pubescence, suggesting that the sf3'h1 gene is involved in the control of pigmentation in pubescence. We detected a marked decrease in target mRNA, an accumulation of short interfering RNAs (siRNAs), and a decrease in quercetin content relative to kaempferol in leaf tissues, indicating that sequence-specific mRNA degradation of the F3'H gene was induced. These results suggest that leaf tissues have a threshold mRNA level of the F3'H gene, which is associated with the occurrence of tawny pigmentation in pubescence. The estimated threshold mRNA level for pigmentation in pubescence was approximately 3% of the steady-state mRNA level of the F3'H gene in greenhouse-grown plants. PMID- 18423788 TI - Protein profiling of single epidermal cell types from Arabidopsis thaliana using surface-enhanced laser desorption and ionization technology. AB - Here, we describe a novel approach for investigating differential protein expression within three epidermal cell types. In particular, 3000 single pavement, basal, and trichome cells from leaves of Arabidopsis thaliana were harvested by glass micro-capillaries. Subsequently, these single cell samples were joined to form pools of 100 individual cells and analyzed using the ProteinChip technology; SELDI: surface-enhanced laser desorption and ionization. As a result, numerous protein signals that were differentially expressed in the three epidermal cell types could be detected. One of these proteins was characterized by tryptical digestion and subsequent identification via tandem quadrupole-time of flight (Q-TOF) mass spectrometry. Down regulation of this sequenced small subunit precursor of ribulose-1,5 bisphosphate carboxylase(C) oxygenase(O) (RuBisCo) in trichome and basal cells indicates the sink status of these cell types that are located on the surface of A. thaliana source leaves. Based on the obtained protein profiles, we suggest a close functional relationship between basal and trichome cells at the protein level. PMID- 18423789 TI - Isolation of cardiac mast cells in experimental Trypanosoma cruzi infection. AB - Mast cells (MC) secrete diverse pre-stored chemical mediators that are pivotal in inflammatory and fibrotic etiologies, such as Trypanosoma cruzi-induced myocardiopathy. However, due to reduced number of cardiac MC, in situ and in vitro identification, and difficult tissue isolation, these cells are rarely addressed. In this work we optimized the identification of cardiac and peritoneal MC and developed an enzymatic method for MC isolation using control and T. cruzi infected mice. MC were identified by: toluidine blue (TB); alcian blue (AB)/safranin (S); AB or a mixed solution composed by AB/S/TB. Previous evaluations of cardiac MC in T. cruzi infection were based on TB staining and our results using AB/S/TB solution showed an increase in, at least, five times the detection of MC. This mixed solution may improve the identification of MC populations also from skin, mucosa and tissues that are infected by other pathogens or under the influence of chronic inflammation, leading to more precise results. Furthermore, the appropriate combination of samples (frozen/unfixed/thick slices) and staining protocols can assure the best evaluation of MC. We have also isolated cardiac MC using collagenase and developed a highly efficient 60%/70% Percoll-graded protocol that enriched in, at least, 95% the population of cardiac MC. PMID- 18423790 TI - The treatment and prognosis of dermatomyositis: an updated review. AB - Dermatomyositis (DM) is an idiopathic inflammatory myopathy. The mainstay of treatment for DM is oral corticosteroids. However, the dose and length of treatment is debated. Adding to the confusion, there have been no randomized controlled studies comparing the use of various corticosteroid doses and taper rates, and no controlled long-term studies assessing the hypothesis that, unlike systemic lupus erythematous, patients with DM can often achieve long-term remission off therapy. This literature review supports an approach that prednisone should be started at about 1 mg/kg/d, which is then tapered slowly based on the response. As patients respond differently to prednisone, additional therapies may be necessary. When to initiate these therapies requires clinical judgment. In addition, as we learn more about the pathophysiology of DM, newer medications that target specific mechanisms in the immune response may help us better treat the disease. Evidence-based data with long-term follow-up will allow for selection of the best treatment to maximize long-term remission, not simply short-term lowering of the systemic corticosteroid dose. PMID- 18423792 TI - Stycholysin II, a cytolysin from the sea anemone Stichodactyla helianthus promotes higher hemolysis in aged red blood cells. AB - We have investigated the relationship between the status of red blood cells (RBCs) and their susceptibility to toxin sticholysin II (StII) hemolytic activity; we have evaluated this effect in different RBC ensembles, comprising young and old cells, and in cells partially damaged by their pre-exposition to a free radical source. Upon action of StII, young cell populations are less prone to hemolysis than the whole population, while old cell populations and peroxyl oxidized red cells are lysed faster than the whole population. Cell K(+) content was higher in young cells and lower in both senescent cells and in peroxyl damaged cells relative to whole cell population. The relevance of cell K(+) content in St II-induced lysis was shown when external Na(+) was partially replaced by K(+); under this condition, RBC lysed faster in the presence of St II but no difference was observed among young cells, whole cells population and peroxyl-damaged cells; only old cells lysed faster that the whole population, response that can be due to an enhanced St II-induced pore formation as supported by evaluation of St II irreversible binding to RBC. It is concluded that this factor and the amount of intracellular K(+) are the dominant parameters that modulate the resistance of RBC to St II-induced lysis. PMID- 18423793 TI - Identification of a novel S-superfamily conotoxin from vermivorous Conus caracteristicus. AB - Conotoxins have been classified into several different superfamilies based on the highly conserved signal peptide sequences of their precursors. However, little is known about the five disulfide bonds containing S-superfamily conotoxins. Only two S-superfamily conotoxins have been identified but their cDNAs are not reported. In this work, we identified a novel S-superfamily conotoxin ca8a from vermivorous Conus caracteristicus. Its sequence shares no homology with those of two other previously reported toxins of the same superfamily, but they have the same cysteine framework, in particular the CX(3)CXC-CXC-CXCXC pattern at the C terminal part. This implies that these toxins might have the same spatial scaffold, but different local conformation or residue side chains may be the cause of their different biological functions. Furthermore, the cDNA of ca8a was cloned with the RACE method. ca8a has a signal peptide sequence different from those of other conotoxins. This gives a defining feature of S-superfamily conotoxins and led to the cloning of more S-superfamily conotoxins from cone snails of different prey types, which indicates that S-superfamily conotoxins widely exist. These results will certainly enrich our understanding of the highly diversified S-superfamily conotoxins. PMID- 18423794 TI - A multigene family of Heteractis magnificalysins (HMgs). AB - Sea anemones are passive predators. They use their specialized stinging cells (nematocysts) to immobilize any prey that blunders into them. A cnida fires, everting a tubule which delivers toxins that may stick to a prey. These toxins include neurotoxins, cytotoxins, cardiotoxins and haemolysins. Heteractis magnificalysins (HMgs) belong to a family of cytolysins from the sea anemone Heteractis magnifica. HMgs are 19.5kDa basic proteins of 177 amino acids with pI values ranging from 8 to 10. From 52 cloned HMg gene sequences, we showed that HMgs are encoded by a multigene family whose members are highly homologous to each other. HMg genes are intronless, and may have arisen by gene duplication, gene conversion or mutation. By modifying the extraction procedure, we purified more natural HMg proteins from H. magnifica, thus demonstrating that H. magnifica are naturally competent to produce a large number of HMg cytolysins. Native and recombinant HMg proteins differed from each other in their amino acid sequences and biological activities. In each H. magnifica, many cytolysin isoforms are produced. H. magnifica appeared to have evolved a survival mechanism whereby a large number of cytolysins of different biological properties are produced for defense and offence. PMID- 18423795 TI - Various routes of administration of (99m)Tc-labeled synthetic lactoferrin antimicrobial peptide hLF 1-11 enables monitoring and effective killing of multidrug-resistant Staphylococcus aureus infections in mice. AB - The synthetic antimicrobial peptide representative of the first 11 N-terminal amino acids of human lactoferrin (hLF 1-11) kills multidrug-resistant Staphylococcus aureus (MRSA). This study displays antimicrobial activity of hLF 1 11, via various routes of administration, against MRSA infections in mice. Radiolabeling hLF 1-11 with technetium-99m ((99m)Tc-hLF 1-11) enables scintigraphic monitoring directly after administration. (99m)Tc-hLF 1-11 was taken up by the gall bladder, intestines, and kidneys. Most of the radioactivity was captured in the urinary bladder and about 1% of the injected dose accumulated into infected thigh muscles. At 2 or 24h after either intravenously, subcutaneously, intraperitoneally, or orally injected a single dose of 0.04 mg/kg hLF 1-11 in mice significantly reduced (20-60 times) the number of viable MRSA. In a dose-response setting in immunocompetent mice maximum bactericidal effects (10,000 times reduction) of intravenously injected (99m)Tc-hLF 1-11 was seen with 40 mg/kg whereas the same dose of orally administered (99m)Tc-hLF 1-11 induced about approximately 100 times reduction. In conclusion, intravenously and orally administrated (99m)Tc-hLF 1-11 accumulates in infected tissues and is highly effective against experimental infections with MRSA. Moreover, scintigraphy is an excellent tool to study the pharmacology of experimental compounds and to determine the uptake in infected tissues. PMID- 18423796 TI - Novel brevinins from Chinese piebald odorous frog (Huia schmackeri) skin deduced from cloned biosynthetic precursors. AB - Antimicrobial peptides represent the most characterized and diverse class of peptides within the defensive skin secretions of anuran amphibians. With an ever expanding database of primary structures, the current accepted rules for nomenclature have become increasingly difficult to apply to peptides whose primary structural attributes are either unique or that fall between those that define existing groups. An additional factor that adds to the confusion is the regular re-classification or revision of existing taxa. In the present study, we have identified five new antimicrobial peptide homologs in the defensive skin secretion of the Chinese piebald odorous frog, Huia schmackeri (formerly Rana (Odorrana) schmackeri), by cloning of their respective biosynthetic precursors. As these peptides are obvious homologs of the brevinin-1 and brevinin-2 families we have named these in accordance: (1) brevinin-1HS1, (2) brevinin-2HS1, (3) brevinin-2HS2, (4) brevinin-2HS3 and (5) brevinin-1HS2. The reasons for adopting these names are discussed. It is clear that with an ever-increasing number of amphibian skin antimicrobial peptides appearing in the literature that a consistent nomenclature scheme needs to be established. PMID- 18423797 TI - Solution structure of urotensin-II receptor extracellular loop III and characterization of its interaction with urotensin-II. AB - Urotensin-II (U-II) is a vasoactive hormone that acts through a G-protein-coupled receptor named UT. Recently, we have shown, using the surface plasmon resonance technology that human U-II (hU-II) interacts with the hUT(281-300) fragment, a segment containing the extracellular loop III (EC-III) and short extensions of the transmembrane domains VI and VII (TM-VI and TM-VII). To further investigate the interaction of UT receptor with U-II, we have determined the solution structure of hUT(281-300) by high-resolution NMR and molecular modeling and we have examined, also using NMR, the binding with hU-II at residue level. In the presence of dodecylphosphocholine micelles, hUT(281-300) exhibited a type III beta-turn (Q285-L288), followed by an -helical structure (A289-L299), the latter including a stretch of transmembrane helix VII. Upon addition of hU-II, significant chemical shift perturbations were observed for residues located just on the N-terminal side of the beta-turn (end of TM-VI/beginning of EC-III) and on one face of the -helix (end of EC-III/beginning of TM-VII). These data, in conjunction with intermolecular NOEs, suggest that the initiation site of EC-III, as well as the upstream portion of helix VII, would be involved in agonist binding and allow to propose points of interaction in the ligand-receptor complex. PMID- 18423798 TI - Characterizing the extracellular and intracellular fluorescent products of activated sludge in a sequencing batch reactor. AB - Three-dimensional excitation-emission-matrix (EEM) fluorescence spectrometry was used to characterize the extracellular and intracellular substances of activated sludge in a sequencing batch reactor (SBR). Parallel factor analysis (PARAFAC) was applied to extract the pure spectra from the overlapped spectra. Three main components, proteins, fulvic- and humic-like substances, were identified from the extracellular substances. Their fluorescence peaks were at an excitation/emission (Ex/Em) of 280/350, 340/400 and 390/450 nm, respectively. The fluorescence of the extracellular proteins had a similar changing pattern with the wastewater chemical oxygen demand, the fulvic-like substance did not vary significantly in a cycle and the humic-like substances accumulated in the substrate uptake phase but decreased later. Proteins and nicotinamide adenine dinucleotide, reduced form (NADH), were identified as the two main intracellular fluorophores, and their fluorescence peaks (Ex/Em) were at 280/340 and 350/450 nm, respectively. The fluorescence intensity scores of the intracellular fluorophores were closely related to the bioreactor performance. Thus, the results of this work provide a foundation for potential utilization of the EEM fluorescence spectroscopy to monitor the activated sludge systems for wastewater treatment. PMID- 18423799 TI - Presence and activity of ammonia-oxidising bacteria detected amongst the overall bacterial diversity along a physico-chemical gradient of a nitrifying wastewater treatment plant. AB - We wished to discover if we could gain greater insights into how biological treatment plants function by contrasting the presence and activity of the most abundant Bacteria in plug flow and completely mixed activated sludge plants. Presence was assessed by amplifying 16S rRNA gene fragments (using PCR) and activity by amplifying native 16S rRNA, using reverse-transcriptase PCR (RT-PCR), using Bacteria-specific primers. The amplified sequences were compared using denaturing gradient gel electrophoresis (DGGE). The plug flow plant exhibited a strong physico-chemical gradient with an initial anoxic zone, whilst the two completely mixed reactors did not. Similarities were observed between the profile of the banding pattern for presence and activity. However, in the plug flow reactor one prominent band was detected in the active population (16S rRNA) but was absent from the corresponding profile of the 16S rRNA gene. Sequencing of this band revealed its identity as a Nitrosomonas-like sequence. The intensity of the 16S rRNA sequenced varied along the physico-chemical gradient of the plug flow reactor in a manner that coincided with the growth of ammonia-oxidising bacteria (AOB) and the loss of ammonia. This band was also absent from the completely mixed reactors, although significant numbers of AOB were detected in all systems ( approximately 10(6)-10(8)cells ml(-1)) by fluorescence in situ hybridisation (FISH). An abundant and highly active AOB population was present in the anoxic zone of the plug-flow reactor where up to 60% of the total ammonia was removed. An examination of nitrogen removal/production rates, together with the above data, reveal that complex nitrogen removal processes occur in this system. These data also enabled the calculation of a specific in situ growth rate for the AOB as 0.12h(-1). PMID- 18423800 TI - Synthesis, cytotoxicity and apoptosis of naphthalimide polyamine conjugates as antitumor agents. AB - Several naphthalimide polyamine conjugates were synthesized and evaluated for in vitro cytotoxicity against human leukemia K562, murine melanoma B16, Chinese hamster ovary CHO cell lines. Both triamine moieties and the length of spacers were crucial in elevating the potency of 1,8-naphthalimide. The typical compounds 5a and 5d exhibited excellent cell selectivity to cancer cells through the human hepatoma BEL-7402 and human normal hepatocyte QSG-7701 screens. In addition, 5d could disturb the cell cycle in B16 cells. The research on caspase activity and cytochrome c indicated that 5d could induce B16 cell apoptosis via both the mitochondrial and membrane death receptor pathways, and the Bcl-2 family numbers were involved in the control of apoptosis. PMID- 18423801 TI - Luminescence chronology of cave sediments at the Atapuerca paleoanthropological site, Spain. AB - Ascertaining the timing of the peopling of Europe, after the first out-of-Africa demographic expansion at the end of the Pliocene, is of great interest to paleoanthropologists. One of the earliest direct evidences for fossil hominins in western Europe comes from an infilled karstic cave site called Gran Dolina at Atapuerca, in a stratum approximately 1.5m below the Brunhes-Matuyama (B-M) geomagnetic boundary (780ka) within lithostratigraphic unit TD6. However, most of the meters of fossil- and tool-bearing strata at Gran Dolina have been difficult to date. Therefore, we applied both thermoluminescence (TL) and infrared stimulated-luminescence (IRSL) multi-aliquot dating methods to fine-silt fractions from sediment samples within Gran Dolina and the nearby Galeria cave site. We also applied these methods to samples from the present-day surface soils on the surrounding limestone hill slopes to test the luminescence-clock-zeroing by-daylight assumption. Within the uppermost 4m of the cave deposits at Gran Dolina, TL and paired TL and IRSL ages range stratigraphically from 198+/-19ka to 244+/-26ka. Throughout Gran Dolina, all luminescence results are stratigraphically self-consistent and, excepting results from two stratigraphic units, are consistent with prior ESR-U-series ages from progressively deeper strata. Thermoluminescence ages culminate at 960+/-120ka approximately 1m below the 780ka B-M boundary. At Galeria, with one exception, TL and IRSL ages range stratigraphically downward from 185+/-26ka to 503+/-95ka at the base of the lowermost surface-inwash facies. These results indicate that TL and (sometimes) IRSL are useful dating tools for karstic inwash sediments older than ca. 100ka, and that a more accurate chronostratigraphic correlation is now possible among the main Atapuerca sites (Gran Dolina, Galeria, Sima de los Huesos). Furthermore, the oldest TL age of ca. 960ka from Gran Dolina, consistent with biostratigraphic and paleomagnetic evidence, implies a probable numeric age of 900-950ka for the oldest hominin remains ( approximately 0.8m below the TL sample). This age window suggests a correspondence to Marine Isotope Stage (MIS) 25, a relatively warm and humid interglaciation. PMID- 18423802 TI - Mitochondrial relationships and divergence dates of the African colobines: evidence of Miocene origins for the living colobus monkeys. AB - The African colobines represent a neglected area of cercopithecid systematics. Resolving the phylogenetic relationships and estimating divergence dates among the living forms will provide insight into the evolution of this group and may shed light upon the evolution of other African primates as well. This is the first molecular assessment of the evolutionary relationships among the modern colobus monkeys, which are comprised of the black-and-white, olive, and red colobus groups. Over 4,000 base pairs of mitochondrial DNA were amplified and sequenced in over 40 colobus monkey individuals incorporating representatives from all commonly recognized species. Gene trees were inferred using maximum likelihood and Bayesian inference, and penalized likelihood was employed to estimate mitochondrial divergence dates among the sampled taxa. The results are congruent with some aspects of previous phylogenetic hypotheses based on morphology and vocalizations, although the relationships among several West and Central African taxa differ to some degree. The divergence date analysis suggests that the black-and-white, olive, and red colobus had diverged from one another by the end of the Miocene, and that by the Plio-Pleistocene many of the species lineages were already present. This demonstrates that the initial extant colobus monkey diversification occurred much earlier than previously thought and was likely part of the same adaptive radiation that produced the diverse colobine taxa seen in the African Plio-Pleistocene fossil record. The lack of early members from the modern lineages in fossiliferous deposits suggests that they resided in part in the forests of Central and West Africa, which also currently harbor the highest levels of colobus monkey diversity. These forests should not be ignored in models of Plio-Pleistocene human and nonhuman primate evolution. PMID- 18423803 TI - Canine MDCK cell lines are refractory to infection with human and mouse prions. AB - Influenza vaccine production in embryonated eggs is associated with many disadvantages, and production in cell culture systems is a viable alternative. Madin Darby canine kidney (MDCK) cells are permissive for a variety of orthomyxoviruses and have proven particularly suitable for vaccine mass production. However, mammalian cells harboring the Prnp gene can theoretically acquire prion infections. Here, we have attempted to infect MDCK cells and substrains thereof with prions. We found that MDCK cells did not produce any protease-resistant PrP(Sc) upon exposure to brain homogenates derived from humans suffering from Creutzfeldt-Jakob disease (CJD) or from mice infected with Rocky Mountain Laboratory (RML) scrapie prions. Further, transmission of MDCK lysates to N2aPK1 cells did not induce formation of PrP(Sc) in the latter. PrP(C) biogenesis and processing in MDCK cells were similar to those of prion-sensitive N2aPK1 cells. However, steady-state levels of PrP(C) were very low, and PrP(C) did not partition with detergent-resistant membranes upon density gradient analysis. These factors may account for their resistance to infection. Alternatively, prion resistance may be related to the specific sequence of canine Prnp, as suggested by the lack of documented prion diseases in dogs. PMID- 18423804 TI - Addition of CpG ODN to recombinant Pseudomonas aeruginosa ExoProtein A conjugates of AMA1 and Pfs25 greatly increases the number of responders. AB - Both the blood-stage protein apical membrane antigen 1 (AMA1) and the 25-kDa sexual-stage protein (Pfs25) of Plasmodium falciparum are two leading candidates in malarial vaccine development. We have previously demonstrated that conjugation of these malarial antigens to recombinant Pseudomonas aeruginosa ExoProtein A (rEPA) significantly increased the mean-specific functional antibody responses in mice; however, some mice responded poorly and were unable to demonstrate a functional response. We hypothesized that the immunogenicities of these two malarial antigens could be further enhanced by the inclusion of a CpG oligodeoxynucleotide in the formulation. Mice were immunized with either rEPA conjugated or unconjugated AMA1 and Pfs25 formulated on Alhydrogel with or without the addition of CPG 7909. Mice received the formulations on days 0 and 28, and mouse sera were collected on day 42. ELISA analyses on these sera showed that the addition of CPG 7909 to AMA1-rEPA and Pfs25-rEPA formulated on Alhydrogel induced significantly higher mean antibody titers than the formulations without CPG 7909, and led to a mixed Th1/Th2 response as demonstrated by the production of mouse IgG1 and IgG2a subclasses. The presence of CPG 7909 in the formulations of both conjugated antigens greatly increased the proportion of responders with antibody titers sufficient to inhibit blood-stage parasite growth in vitro or block transmission of sexual-stage parasites to mosquitoes. The results obtained in this study indicate the potential use of a combination strategy to increase the number of responders to malarial antigens in humans. PMID- 18423805 TI - Livestock vaccine adoption among poor farmers in Bolivia: remembering innovation diffusion theory. AB - The paper explores the low uptake of livestock vaccination among poor farming communities in Bolivia utilising core elements of the original innovation diffusion theory. Contrary to the recent literature, we found that vaccination behaviour was strongly linked to social and cultural, rather than economic, drivers. While membership in a group increased uptake, the 'hot' and 'cold' distinctions which dictate health versus illness within Andean cosmology also played a role, with vaccination viewed as a means of addressing underlying imbalances. We concluded that uptake of livestock vaccination was unlikely to improve without knowledge transfer that acknowledges local epistemologies for livestock disease. PMID- 18423807 TI - Reduction in beta2-microglobulin with super-flux versus high-flux dialysis membranes: results of a 6-week, randomized, double-blind, crossover trial. AB - BACKGROUND: Uremic toxicity is a major concern in the dialysis population. There is keen interest in techniques that increase the removal of larger uremic molecules. We examined the short-term impact of a new, more porous, super-flux Helixone membrane (FX-E) versus the conventional high-flux Helixone membrane (FX 60) on beta(2)-microglobulin (beta2M) reduction and nutritional and inflammatory parameters. STUDY DESIGN: Randomized, double blind, crossover, pilot trial. SETTING & PARTICIPANTS: A single freestanding dialysis center. 30 stable hemodialysis patients. INTERVENTION: Patients were treated with FX-60 and FX-E membranes for a treatment period of 6 weeks each, with a 2-week washout period in between. OUTCOME & MEASUREMENTS: Primary outcome was change in beta2M concentrations from baseline to end of treatment. Serum samples were obtained predialysis and postdialysis at 0, 2, and 6 weeks, and dialysate albumin samples were collected continuously throughout dialysis sessions. RESULTS: Mean postdialysis beta2M concentrations at the end of 6 weeks of treatment were 6.73 mg/L for FX-E versus 8.22 mg/L for FX-60, which was significantly lower overall by 0.69 mg/L (95% confidence interval [CI], -1.09 to -0.29; P = 0.001). beta2M reduction ratios were greater overall with FX-E by 4.83% (95% CI, 2.78 to 6.89; P < 0.001), with mean values of 57% for FX-60 versus 66% for FX-E at the end of treatment. Median dialysate albumin loss with FX-E was 1.23 g (range, 0.22 to 4.83 g) compared with 0.17 g (range, 0.0017 to 2.69 g) with FX-60, which was greater by 1.52 g (95% CI, 1.11 to 1.93; P < 0.001). Serum albumin concentrations were slightly lower with FX-E by 0.1 g/dL (0.55 g/L; 95% CI, -1.04 to -0.07; P = 0.03), but prealbumin concentrations were not significantly different at 8.53 mg/L (95% CI, -23.76 to 6.71; P = 0.3). There were no differences in inflammatory cytokine concentrations or small-solute removal. LIMITATIONS: Short-term pilot study. CONCLUSION: In this stable dialysis population, removal of beta2M was more efficient with the Helixone super-flux FX-E membrane, with only a small decrease in albumin concentrations despite increased albumin loss. Large trials with longer treatment periods are required to evaluate the impact of the FX-E membrane on clinical outcomes. PMID- 18423808 TI - Two rare forms of renal allograft glomerulopathy during cytomegalovirus infection and treatment. PMID- 18423809 TI - A 1-year randomized trial of calcium acetate versus sevelamer on progression of coronary artery calcification in hemodialysis patients with comparable lipid control: the Calcium Acetate Renagel Evaluation-2 (CARE-2) study. AB - BACKGROUND: Previous clinical trials showed that progression of coronary artery calcification (CAC) may be slower in hemodialysis patients treated with sevelamer than those treated with calcium-based phosphate binders. Because sevelamer decreases low-density lipoprotein cholesterol (LDL-C) levels, we hypothesized that intensive lowering of LDL-C levels with atorvastatin in hemodialysis patients treated with calcium acetate would result in CAC progression rates similar to those in sevelamer-treated patients. STUDY DESIGN: Randomized, controlled, open-label, noninferiority trial with an upper bound for the noninferiority margin of 1.8. SETTING & PARTICIPANTS: 203 prevalent hemodialysis patients at 26 dialysis centers with serum phosphorus levels greater than 5.5 mg/dL, LDL-C levels greater than 80 mg/dL, and baseline CAC scores of 30 to 7,000 units assessed by means of electron-beam computed tomography. INTERVENTIONS: 103 patients were randomly assigned to calcium acetate, and 100 patients to sevelamer for 12 months to achieve phosphorus levels of 3.5 to 5.5 mg/dL. Atorvastatin was added to achieve serum LDL-C levels less than 70 mg/dL in both groups. OUTCOMES & MEASUREMENTS: The primary end point was change in CAC score assessed by means of electron-beam computed tomography. RESULTS: After 12 months, mean serum LDL-C levels decreased to 68.8 +/- 22.0 mg/dL in the calcium-acetate group and 62.4 +/- 23.0 mg/dL in the sevelamer group (P = 0.3). Geometric mean increases in CAC scores were 35% in the calcium-acetate group and 39% in the sevelamer group, with a covariate-adjusted calcium acetate-sevelamer ratio of 0.994 (95% confidence interval, 0.851 to 1.161). LIMITATIONS: Treatment assignment was not blinded. The 1.8 a priori margin is large, CAC is a surrogate outcome, duration of treatment was short, and dropout rate was high. CONCLUSIONS: With intensive lowering of LDL C levels for 1 year, hemodialysis patients treated with either calcium acetate or sevelamer experienced similar progression of CAC. PMID- 18423806 TI - Base excision repair, aging and health span. AB - DNA damage and mutagenesis are suggested to contribute to aging through their ability to mediate cellular dysfunction. The base excision repair (BER) pathway ameliorates a large number of DNA lesions that arise spontaneously. Many of these lesions are reported to increase with age. Oxidized guanine, repaired largely via base excision repair, is particularly well studied and shown to increase with age. Spontaneous mutant frequencies also increase with age which suggests that mutagenesis may contribute to aging. It is widely accepted that genetic instability contributes to age-related occurrences of cancer and potentially other age-related pathologies. BER activity decreases with age in multiple tissues. The specific BER protein that appears to limit activity varies among tissues. DNA polymerase-beta is reduced in brain from aged mice and rats while AP endonuclease is reduced in spermatogenic cells obtained from old mice. The differences in proteins that appear to limit BER activity among tissues may represent true tissue-specific differences in activity or may be due to differences in techniques, environmental conditions or other unidentified differences among the experimental approaches. Much remains to be addressed concerning the potential role of BER in aging and age-related health span. PMID- 18423811 TI - The history of renal transplantation in the Arab world: a view from Saudi Arabia. AB - The first successful renal transplantation in the Arab world took place in Jordan in 1972. Surprisingly, the kidney transplanted was from a non-heart-beating deceased donor. Many Arab countries followed suit, starting their transplantation programs in the 1970s and 1980s, but all were from living related donors. Very few Arab countries managed to start deceased donor programs, notable among which is the Kingdom of Saudi Arabia. Religion has an important part in personal life and government legislation in the Arab world; thus, organ procurement and transplantation had to wait for religious edicts (fatwas) to be passed about the permissibility of organ donation and brain death diagnosis before starting transplantation activities. In Saudi Arabia, the renal transplantation service went through several developmental phases, culminating in the establishment of the Saudi Center for Organ Transplantation, which has become the prototype of a successful multiorgan procurement center to be emulated by Arab and Muslim countries. The story of transplantation in the Arab world is intertwined and shaped by the prevailing socioeconomic and health indicators in the different countries. It also is the story of hard-working pioneers and of human endeavor against adversity, exemplified by 2 of the pioneers having received organ transplants. Arab countries have had more than their fair share of strife and wars, and this has impacted on transplantation services and programs. PMID- 18423810 TI - Relationship of impaired olfactory function in ESRD to malnutrition and retained uremic molecules. AB - BACKGROUND: Olfactory function is impaired in patients with end-stage renal disease (ESRD) and may contribute to uremic anorexia. Only limited correlations of olfactory function and nutritional status were reported. This study examines the relationship of impaired olfactory function to malnutrition and levels of the retained uremic solutes monomethylamine, ethylamine, indoxyl sulfate, and P cresol sulfate. STUDY DESIGN: Cross-sectional observational study. SETTING & PARTICIPANTS: 31 stable maintenance hemodialysis patients from an urban outpatient dialysis unit and 18 people with normal renal function participated. PREDICTOR: Nutritional status assigned by using Subjective Global Assessment (SGA) score; SGA score of 7 indicates normal nutritional status; SGA score of 5 to 6, mild malnutrition; and SGA score of 3 to 4, moderate malnutrition. OUTCOMES & MEASUREMENTS: The primary outcome is olfactory function, assessed using the University of Pennsylvania Smell Identification Test. Levels of retained uremic solutes were measured from a predialysis serum sample. Demographic data and laboratory values for nutritional status, adequacy of dialysis, and inflammation were collected. RESULTS: Mean smell scores were 34.9 +/- 1.4 for controls, 33.5 +/- 3.3 for patients with SGA score of 7, 28.3 +/- 5.8 for patients with SGA score of 5 to 6, and 27.9 +/- 4.4 for patients with SGA score of 3 to 4 (P < 0.001 comparing healthy patients with all patients with ESRD). There was no difference in mean smell scores for healthy controls and patients with SGA score of 7. However, patients with lower smell scores had significantly lower SGA scores (P = 0.02) and higher C-reactive protein levels (P = 0.02). Neither smell score nor nutritional status was associated with levels of retained uremic solutes. LIMITATIONS: Small sample size, only cross-sectional associations can be described. CONCLUSIONS: Our results suggest an association between poor nutritional status and impaired olfactory function in patients with ESRD. Additional research is needed to discover the uremic toxins mediating these processes. PMID- 18423812 TI - Triple pharmacological blockade of the renin-angiotensin-aldosterone system in nondiabetic CKD: an open-label crossover randomized controlled trial. AB - BACKGROUND: Agents inhibiting the renin-angiotensin-aldosterone (RAAS) system have an important role in slowing the progression of chronic kidney disease. We evaluated the hypothesis that the addition of an aldosterone receptor antagonist to an angiotensin-converting enzyme (ACE) inhibitor and angiotensin II type 1 (AT 1) receptor blocker (ARB) (triple RAAS blockade) may provide an additional benefit compared with an ACE inhibitor and ARB (double RAAS blockade). DESIGN: Randomized open controlled crossover study. SETTING & PARTICIPANTS: 18 whites (7 women, 11 men) from the Outpatient Department of Nephrology with chronic nondiabetic proteinuric kidney diseases, mean age 42.4 +/- 1.9 years (SEM). INTERVENTIONS: In the 8-week run-in period, all participants received the ACE inhibitor cilazapril (5 mg), the ARB telmisartan (80 mg), and the diuretic hydrochlorothiazide (12.5 mg) as double RAAS blockade to achieve the target blood pressure of less than 130/80 mm Hg. Participants were then randomly assigned to 2 treatment sequences, either the addition of spironolactone (25 mg) (triple RAAS blockade) through 8 weeks followed by double RAAS blockade through 8 weeks (sequence 1) or double RAAS blockade followed by triple RAAS blockade (sequence 2). MAIN OUTCOME MEASURES: 24-hour urine protein excretion (primary end point) and markers of tubular injury and fibrosis (secondary end points). Analysis was performed using analysis of variance for repeated measurements. RESULTS: At baseline, mean serum creatinine level was 1.16 +/- 0.09 mg/dL (103 +/- 8 micromol/L), estimated glomerular filtration rate was 107.8 mL/min (95% confidence interval, 93 to 140.9 [1.8 mL/s; 95% confidence interval, 1.55 to 2.35; Cockcroft-Gault formula), and 24-hour mean proteinuria was 0.97 +/- 0.18 g. Mean urine protein excretion was 0.7 g/24 h (95% confidence interval, 0.48 to 0.92) less after triple RAAS blockade than after double RAAS blockade (P = 0.01), without change in blood pressure. Urine excretion of N-acetyl-beta-d glucosaminidase (P = 0.02) and amino-terminal propeptide of type III procollagen (P = 0.05) also significantly decreased. Potassium levels increased significantly after triple therapy (P = 0.02). However, no patient was withdrawn because of adverse effects. LIMITATIONS: Absence of blinding, small sample size, short treatment period, absence of histological assessment. CONCLUSIONS: Administration of an aldosterone receptor antagonist in addition to double RAAS blockade with an ACE inhibitor and ARB may slow the progression of chronic kidney disease. Additional studies are necessary to confirm this result. PMID- 18423813 TI - Kidney function and risk factors for left ventricular hypertrophy in untreated uncomplicated essential hypertension. AB - BACKGROUND: Left ventricular (LV) hypertrophy and decreased kidney function are well-established cardiovascular risk factors in hypertensive patients. STUDY DESIGN: We investigated the relationship between creatinine level, creatinine clearance, and estimated glomerular filtration rate (eGFR) with LV mass (LVM) in a cross-sectional study. PREDICTORS: eGFR and serum creatinine level. OUTCOME: LVM index (LVMI). SETTING & PARTICIPANTS: 400 patients with untreated uncomplicated essential hypertension. MEASUREMENTS: LVMI, eGFR (Modification of Diet in Renal Disease Study equation), Framingham risk factors, and a series of specific risk factors, ie, endothelial function (acetylcholine [ACh]-stimulated forearm blood flow [FBF]), insulin sensitivity (Homeostatic Model Assessment for insulin resistance [HOMA-R] index), C-reactive protein (CRP), and uric acid. RESULTS: Both eGFR and creatinine level were significantly related to LVMI (r = 0.34 and r = 0.35; P < 0.001). In a multiple regression model adjusting for Framingham risk factors, eGFR was independently associated with LVMI. However, this association, although highly significant, lost substantial strength after adjustment for such specific risk factors as HOMA-R index, ACh-stimulated FBF, CRP level, and uric acid level. eGFR interacted with insulin resistance in explaining the variability in LVMI (P = 0.007). LIMITATIONS: The cross-sectional nature of this study precludes cause-effect conclusions. CONCLUSIONS: Independently of other risk factors, decreased kidney function contributes to explain the variability in LVMI in patients with untreated uncomplicated essential hypertension. This association is attributable in part to the link between eGFR and such specific risk factors as HOMA-R index, ACh-stimulated FBF, CRP level, and uric acid level. Decreased kidney function and insulin resistance interact in explaining the variability in LVMI in these patients. PMID- 18423814 TI - Obesity-related glomerulopathy in China: a case series of 90 patients. AB - BACKGROUND: The epidemic of obesity has been paralleled by an increase in the incidence of chronic kidney disease. However, epidemiological data for obesity related glomerulopathy (ORG) from developing countries, including China, are very limited. STUDY DESIGN: Case series. ORG defined as body mass index (BMI) of 28.0 kg/m(2) or greater; urinary protein excretion of 0.4 g/24 h or greater, and glomerulomegaly (glomerular volume > 3.27 x 10(6) microm(3)) with or without focal segmental glomerulosclerosis (FSGS). SETTING & PARTICIPANTS: 10,093 renal biopsy samples from patients obtained from February 2002 to November 2006 at the Research Institute of Nephrology, Nanjing University School of Medicine, China. PREDICTOR: Obesity defined as a BMI of 28.0 kg/m(2) or greater. Subjects were divided into 3 groups: mild-obesity group with BMI of 28.0 to less than 30 kg/m(2), moderate-obesity group with BMI of 30 to less than 35 kg/m(2), and severe-obesity group with BMI of 35 kg/m(2) or greater. OUTCOMES & MEASUREMENTS: Clinicoepidemiological and histopathologic characteristics of patients with ORG at the time of biopsy were described separately. RESULTS: ORG was observed in 90 biopsy specimens (0.89%); frequency increased from 0.62% to 1.0% during the last 5 years (P = 0.02). Mean age was 37.5 +/- 9.3 (SD) years, 67% were men, mean BMI was 31.2 +/- 3.3 kg/m(2), waist circumference was 103 cm (range, 89.4 to 124 cm) in men and 96.5 cm (range, 88.5 to 113 cm) in women, waist-hip ratio was 0.95 +/- 0.07, and 100% had visceral obesity. Of the total, 49%, 37%, and 14% had mild, moderate, and severe obesity, respectively. Mean urinary protein excretion of subjects was 1.48 +/- 1.2 g/24 h; 51%, 39%, and 10% had proteinuria with protein of 0.4 to 1.0, 1.0 to 3.5, and greater than 3.5 g/d, respectively. Mean measured creatinine clearance (Ccr) was 109 +/- 32.2 mL/min/1.73 m(2), with 42%, 36%, and 22% with a Ccr greater than 120, 90 to 120, and less than 90 mL/min/1.73 m(2), respectively. Glucose dysmetabolism, insulin resistance, dyslipidemia, and hypertension were observed in 77%, 88%, 76%, and 63% of patients, respectively. FSGS was observed in 70%. Mean foot-process width was 534 +/- 176 nm. Foot process fusion was seen in 36% of patients. Greater BMI was associated with greater proteinuria (P < 0.02), greater Ccr (P < 0.03), and greater foot-process width (P < 0.04). LIMITATIONS: Inability to compute prevalence or incidence from case series. BMI was calculated at time of renal biopsy. CONCLUSIONS: Most patients with ORG had mild obesity, visceral obesity, minor proteinuria, preserved Ccr, and FSGS. PMID- 18423815 TI - Mutations in proteins of the alternative pathway of complement and the pathogenesis of atypical hemolytic uremic syndrome. AB - Atypical hemolytic uremic syndrome is associated with mutations in the complement proteins factor H, factor I, factor B, C3, or membrane cofactor protein in about 50% of patients. The evolution and prognosis of the disease in patients carrying mutations in factor H is particularly poor, and renal transplantation most often fails because of recurrence of the disease in the graft. The risk of rapid loss of renal function in patients with functional mutations in factor H requires that effective treatment be initiated as soon as possible, but identification of these patients relies on genetic studies that are time consuming. We describe a case in which an in vitro hemolytic assay proved useful for rapidly assessing factor H dysfunction and for testing whether this dysfunction could be corrected with fresh frozen plasma. In the context of this case, we summarize recent advances in understanding the molecular mechanisms contributing to atypical hemolytic uremic syndrome, including descriptions of DNA- and protein-based analysis. We conclude that functional analysis of factor H should help rationalize the plasma treatment of patients with atypical hemolytic uremic syndrome. PMID- 18423816 TI - Development and application of an oligonucleotide microarray and real-time quantitative PCR for detection of wastewater bacterial pathogens. AB - Conventional microbial water quality test methods are well known for their technical limitations, such as lack of direct pathogen detection capacity and low throughput capability. The microarray assay has recently emerged as a promising alternative for environmental pathogen monitoring. In this study, bacterial pathogens were detected in municipal wastewater using a microarray equipped with short oligonucleotide probes targeting 16S rRNA sequences. To date, 62 probes have been designed against 38 species, 4 genera, and 1 family of pathogens. The detection sensitivity of the microarray for a waterborne pathogen Aeromonas hydrophila was determined to be approximately 1.0% of the total DNA, or approximately 10(3)A. hydrophila cells per sample. The efficacy of the DNA microarray was verified in a parallel study where pathogen genes and E. coli cells were enumerated using real-time quantitative PCR (qPCR) and standard membrane filter techniques, respectively. The microarray and qPCR successfully detected multiple wastewater pathogen species at different stages of the disinfection process (i.e. secondary effluents vs. disinfected final effluents) and at two treatment plants employing different disinfection methods (i.e. chlorination vs. UV irradiation). This result demonstrates the effectiveness of the DNA microarray as a semi-quantitative, high throughput pathogen monitoring tool for municipal wastewater. PMID- 18423817 TI - Mathematical modeling of first flush in highway storm runoff using genetic algorithm. AB - A mechanistic model was developed to predict the highway runoff pollutographs during precipitation events. Pollutants were assumed to be in two phases, attached to the pavement surface and mobile in the runoff water. Detachment and reattachment of contaminants were considered as rate-limited processes and the detachment rate was assumed to be a function of flow velocity by a power expression. The build-up of pollutants on the surface during the dry period between the storm events was also included in the model. Using measured pollutographs from three highly urbanized highway sites in Los Angeles, California, a hybrid genetic algorithm was used to estimate the model parameters for four different constituents including total suspended solids, oil and grease, dissolved copper, and particulate copper. The model was then validated by predicting pollutographs for the same site for a different monitoring season. Results revealed that estimated model parameters were different for particle associated and dissolved constituents. These findings suggest different mechanisms governing the detachment of particle-associated and dissolved constituents from the surface. The results have also indicated that there is a linear build-up of pollutants during the dry period and the removal of pollutants during dry periods was insignificant. From these findings, it has been concluded that either the removal rate during the dry period is small or it is not proportional to the concentration of pollutants accumulated on the surface. In general, the build-up contaminant concentrations from the build-up model followed the same trend. However, in some cases the estimated and measured pollutograph did not closely match that may be due to some unknown factors affecting the build up rates in the current model. PMID- 18423818 TI - Evidence of temperature-dependent effects on the estrogenic response of fish: implications with regard to climate change. AB - Chemical risk assessment is fraught with difficulty due to the problem of accounting for the effects of mixtures. In addition to the uncertainty arising from chemical-to-chemical interactions, it is possible that environmental variables, such as temperature, influence the biological response to chemical challenge, acting as confounding factors in the analysis of mixture effects. Here, we investigate the effects of temperature on the response of fish to a defined mixture of estrogenic chemicals. It was anticipated that the response to the mixture may be exacerbated at higher temperatures, due to an increase in the rate of physiological processing. This is a pertinent issue in view of global climate change. Fathead minnows (Pimephales promelas) were exposed to the mixture in parallel exposure studies, which were carried out at different temperatures (20 and 30 degrees C). The estrogenic response was characterised using an established assay, involving the analysis of the egg yolk protein, vitellogenin (VTG). Patterns of VTG gene expression were also analysed using real-time QPCR. The results revealed that there was no effect of temperature on the magnitude of the VTG response after 2 weeks of chemical exposure. However, the analysis of mixture effects at two additional time points (24 h and 7 days) revealed that the response was induced more rapidly at the higher temperature. This trend was apparent from the analysis of effects both at the molecular and biochemical level. Whilst this indicates that climatic effects on water temperature are not a significant issue with regard to the long-term risk assessment of estrogenic chemicals, the relevance of short-term effects is, as yet, unclear. Furthermore, analysis of the patterns of VTG gene expression versus protein induction gives an insight into the physiological mechanisms responsible for temperature-dependent effects on the reproductive phenology of species such as roach. Hence, the data contribute to our understanding of the implications of global climate change for wild fish populations. PMID- 18423819 TI - Detoxification and oxidative stress responses along with microcystins accumulation in Japanese quail exposed to cyanobacterial biomass. AB - The cyanobacterial exposure has been implicated in mass mortalities of wild birds, but information on the actual effects of cyanobacteria on birds in controlled studies is missing. Effects on detoxification and antioxidant parameters as well as bioaccumulation of microcystins (MCs) were studied in birds after sub-lethal exposure to natural cyanobacterial biomass. Four treatment groups of model species Japanese quail (Coturnix coturnix japonica) were exposed to controlled doses of cyanobacterial bloom during acute (10 days) and sub chronic (30 days) experiment. The daily doses of cyanobacterial biomass corresponded to 0.2-224.6 ng MCs/g body weight. Significant accumulation of MCs was observed in the liver for both test durations and slight accumulation also in the muscles of the highest treatment group from acute test. The greatest accumulation was observed in the liver of the highest treatment group in the acute test reaching average concentration of 43.7 ng MCs/g fresh weight. The parameters of detoxification metabolism and oxidative stress were studied in the liver, heart and brain. The cyanobacterial exposure caused an increase of activity of cytochrome P-450-dependent 7-ethoxyresorufin O-deethylase representing the activation phase of detoxification metabolism. Also the conjugation phase of detoxification, namely the activity of glutathione-S transferase, was altered. Cyanobacterial exposure also modulated oxidative stress responses including the level of glutathione and activities of glutathione related enzymes and caused increase in lipid peroxidation. The overall pattern of detoxification parameters and oxidative stress responses clearly separated the control and the lowest exposure group from all the higher exposed groups. This is the first controlled study documenting the induction of oxidative stress along with MCs accumulation in birds exposed to natural cyanobacterial biomass. The data also suggest that increased activities of detoxification enzymes could lead to greater biotransformation and elimination of the MCs at the longer exposure time. PMID- 18423820 TI - Exposure, epidemiology and human cancer incidence of naphthalene. AB - This report provides a summary of deliberations conducted under the charge for members of Module B participating in the Naphthalene State-of-the-Science Symposium (NS(3)), Monterey, CA, October 9-12, 2006. The panel's charge was to derive consensus estimates of human exposure to naphthalene under various conditions, cancer incidence plausibly associated with these exposures, and identify quintessential research that could significantly reduce or eliminate material uncertainties to inform human cancer risk assessment. Relying in large part on a commissioned paper [Price, P.S., Jayjock, M.A., 2008. Available data on naphthalene exposures: strengths and limitations, in this issue], exposure levels were estimated for background (0.0001-0.003 microg/m(3)), ambient air (0.001-1.0 microg/m(3)), vehicles (0.003-3.0 microg/m(3)), residences (0.1-10 microg/m(3)), mothball use (on-label: 1-100 microg/m(3); off-label: 10-100 microg/m(3)), and occupational (low: 3-100 microg/m(3); high: 30-1,000 microg/m(3)). There have been few published reports of human cancer associated with naphthalene exposure. Several research projects are suggested that could reduce uncertainty in our understanding of human exposure. Using best scientific judgment, it is reasonably certain that the largest non-occupational exposures to naphthalene are indoor/residential exposures, particularly in households that use naphthalene based products such as mothballs. However, even the highest of these exposures is likely to fall one or more orders of magnitude below moderate or high-level occupational exposure levels experienced by the few known cohorts exposed occupationally to naphthalene alone or as part of chemical mixtures such as jet fuel. PMID- 18423821 TI - Dose metrics in the acquisition of skin sensitization: thresholds and importance of dose per unit area. AB - Allergic contact dermatitis is a common occupational and environmental health problem and many hundreds of chemicals have been implicated as skin sensitizers. Sensitization is acquired following topical exposure to a contact allergen and induction of a cutaneous immune response of an appropriate magnitude. For effective assessment and management of human health risks there is a need to appreciate the dose metrics that drive the induction of skin sensitization. The available evidence suggests that under most normal conditions of exposure it is the dose per unit area of chemical that has over-riding impact on the effectiveness of sensitization. The exception to this rule is when the area of the application site drops below a certain critical level. Here we review in detail the evidence which supports dose per unit area as being the critical exposure metric in the induction of skin sensitization, and the mechanistic bases for this relationship. PMID- 18423822 TI - Introduction to the Biomonitoring Equivalents Pilot Project: development of guidelines for the derivation and communication of Biomonitoring Equivalents. PMID- 18423823 TI - Dichloroacetate induces apoptosis in endometrial cancer cells. AB - PURPOSE: A recent landmark study demonstrated that Dichloroacetate (DCA) treatment promoted apoptosis in lung, breast, and glioblastoma cancer cell lines by shifting metabolism from aerobic glycolysis to glucose oxidation coupled with NFAT-Kv1.5 axis remodeling. The objective of this study was to determine whether DCA induces apoptosis in endometrial cancer cells and to assess apoptotic mechanism. METHODS: A panel of endometrial cancer cell lines with varying degrees of differentiation was treated with DCA and analyzed for apoptosis via flow cytometry. Biological correlates such as gene expression, intracellular Ca(2+), and mitochondrial membrane potential were examined to assess apoptotic mechanism. RESULTS: Initiation of apoptosis was observed in five low to moderately invasive cancer cell lines including Ishikawa, RL95-2, KLE, AN3CA, and SKUT1B while treatment had no effect on non-cancerous 293T cells. Two highly invasive endometrial adenocarcinoma cell lines, HEC1A and HEC1B, were found to be resistant to DCA-induced apoptosis. Apoptotic responding cell lines had a significant increase in early and late apoptotis, a decrease in mitochondrial membrane potential, and decreased Survivin transcript abundance, which are consistent with a mitochondrial-regulated mechanism. DCA treatment decreased intracellular calcium levels in most apoptotic responding cell lines which suggests a contribution from the NFAT-Kv1.5-mediated pathway. DCA treatment increased p53 upregulated modulator of apoptosis (PUMA) transcripts in cell lines with an apoptotic response, suggesting involvement of a p53-PUMA-mediated mechanism. CONCLUSIONS: Dichloroacetate effectively sensitizes most endometrial cancer cell lines to apoptosis via mitochondrial, NFAT-Kv1.5, and PUMA-mediated mechanisms. Further investigation of the cancer therapeutic potential of DCA is warranted. PMID- 18423824 TI - Implications of EGFR inhibition in ovarian cancer cell proliferation. AB - OBJECTIVES: Epidermal Growth Factor Receptor (EGFR) is one of the four members of the Human Epidermal Receptor (HER) family and is over-expressed in multiple malignancies. EGFR over-expression in ovarian cancer has been associated with poor prognosis. Targeted inhibition of EGFR via its tyrosine kinase domain is a successful treatment in lung cancer. Our objective was to correlate EGFR over expression and growth inhibition, by EGF receptor inhibitors, in ovarian cancer. MATERIALS AND METHODS: HER expression in nine epithelial ovarian cancer cell lines and one lung cancer cell line was determined by Western blot analysis. EGFR phosphorylation sites were analyzed and DNA sequencing was performed. Cell proliferation assays were performed in the presence of the tyrosine kinase inhibitor, gefitinib, and the EGFR monoclonal antibody, cetuximab. Inhibitory concentrations of 50% of these therapies were determined and compared across all cell lines. The lung cancer cell line, HCC827, was used as a control. RESULTS: Four of nine (44%) ovarian cancer cell lines and the control lung cancer cell line expressed EGFR. These same cell lines showed a common phosphorylated residue at position 992, while other residues were variably phosphorylated. All but one cell line expressed at least one HER family member. Mutational analysis of the ovarian cancer cell lines showed no mutations in EGFR exons 18-21. Cell proliferation assays using gefitinib and cetuximab showed minimal response in the ovarian cancer cell lines when compared to the control HCC827, but relative sensitivity compared to the one cell line that had no HER family expression. CONCLUSIONS: Ovarian cancer cell lines show variable expression of activated EGFR. EGFR inhibition alone, in ovarian tumors that lack a tyrosine kinase mutation or over-express EGFR is unlikely to result in clinical response. PMID- 18423825 TI - Stigma, ethics and policy: a commentary on Bayer's "Stigma and the ethics of public health: Not can we but should we". PMID- 18423826 TI - Illness experience in fibromyalgia syndrome: a metasynthesis of qualitative studies. AB - There is growing interest in synthesizing qualitative research. Despite certain philosophical and methodological difficulties, such syntheses are potentially useful in enriching the insights of empirical qualitative work in a particular area. This paper reports an interpretive review of research into the subjective experience of fibromyalgia syndrome (FMS), utilizing principles of metasynthesis. Twenty-three separate studies were identified. Each study was evaluated using methodological criteria to provide a context for interpretation of substantive findings. Principal findings were extracted and synthesized under four broad categories: experience of symptoms, search for diagnosis, legitimacy, and coping. Our findings re-emphasised the point that pain in FMS is ambiguous and invisible, raising questions of credibility and legitimacy. People with a diagnosis of FMS appear to frame the experience of symptoms within the biomedical model, where FMS is viewed as an organic entity potentially identifiable through biomedical tests. The subjective meaning and perceived legitimacy of the diagnostic label appear to be important factors in the subjective experience of FMS. Coping strategies adopted can be subsumed under Mannerkorpi, K., Kroksmark, T., Ekdahl, C. [1999. How patients with fibromyalgia experience their symptoms in everyday life. Physiotherapy Research International, 4(2), 110-122.] notions of 'struggling', 'adapting', 'in despair' and 'giving up'. Most studies had at least one identified methodological shortcoming, though it is not straightforward to identify the significance of such shortcomings. We conclude that there is scope for further research into the subjective experience of FMS, and into the methodology of metasynthesis, especially in relation to methodological appraisal. PMID- 18423827 TI - Transitions into informal caregiving and out of paid employment of women in their 50s. AB - Data from the Australian Longitudinal Study on Women's Health were used to study the order of events leading to informal caregiving and changes in labour force participation in mid-aged women, taking into account health and socioeconomic status. This analysis included 9857 women who responded to the third (2001) and fourth (2004) surveys and provided data for the caring and employment variables used. Caring was defined as providing care for an ill, frail or disabled person at least 7h/wk. Between 2001 and 2004, the proportion of women caring increased from 12 to 14%. Paid employment participation decreased from 67 to 62% in 2004. Logistic regression model results show that taking up caring between 2001 and 2004 was not statistically significantly associated with employment status in 2001. Among women who took up caring, however, hours spent in paid employment in 2001 was negatively associated with hours spent caring in 2004. Amongst women working in 2001, taking up caring between 2001 and 2004 was associated with reduced participation in paid employment. In conclusion, among mid-aged women, transitions into caregiving were irrespective of time spent in paid employment, but were followed by a decrease in labour force participation. Policies could aim to support continuing labour force participation during caregiving by creating flexible working arrangements; re-employment programs could support women who quit work in getting back to paid employment after a period of caregiving. PMID- 18423828 TI - Church ladies, good girls, and locas: stigma and the intersection of gender, ethnicity, mental illness, and sexuality in relation to HIV risk. AB - Inner city women with severe mental illness may carry multiple stigmatized statuses. In some contexts these include having a mental illness, being a member of an ethnic minority group, being an immigrant, being poor, and being a woman who does not live up to gendered expectations. These potentially stigmatizing identities influence both the way women's sexuality is viewed and their risk for HIV infection. This qualitative study applies the concept of intersectionality to facilitate understanding of how these multiple identities intersect to influence women's sexuality and HIV risk. We report the firsthand accounts of 24 Latina women living with severe mental illness in New York City. In examining the interlocking domains of these women's sexual lives, we find that the women seek identities that define them in opposition to the stigmatizing label of "loca" (Spanish for crazy) and bestow respect and dignity. These identities have unfolded through the additional themes of "good girls" and "church ladies". Therefore, in spite of their association with the "loca", the women also identify with faith and religion ("church ladies") and uphold more traditional gender norms ("good girls") that are often undermined by the realities of life with a severe mental illness and the stigma attached to it. However, the participants fall short of their gender ideals and engage in sexual relationships that they experience as disempowering and unsatisfying. The effects of their multiple identities as poor Latina women living with severe mental illness in an urban ethnic minority community are not always additive, but the interlocking effects can facilitate increased HIV risks. Interventions should acknowledge women's multiple layers of vulnerability, both individual and structural, and stress women's empowerment in and beyond the sexual realm. PMID- 18423829 TI - Fluoxetine alters feeding behavior and leptin levels in chronically-stressed rats. AB - Stress-induced alterations in feeding behavior are sexually dimorphic and have been related to changes in monoamine levels. Fluoxetine is commonly used as an antidepressant and has also been suggested as an adjunct to other strategies to treat obese individuals. Leptin may interact with stress hormones and with the brain serotonergic system, possibly affecting the feeding behavior of stressed rats. The aim of this study is to evaluate the interaction between chronic fluoxetine treatment and leptin levels in adult female Wistar rats submitted to chronic variable stress. After 30 days of stress, control and stressed groups were subdivided into two groups that received daily injections of vehicle or fluoxetine (8 mg/kg, i.p.). Body weight was evaluated before and after fluoxetine treatment. The animals gained weight with time, signifying that there is a difference in weight gain over time when fluoxetine-treated animals are, or not, subjected to the stress model. Both fluoxetine and stress induced a decrease in sweet food consumption. On the 60th day of fluoxetine treatment, leptin levels were decreased in fluoxetine-treated animals and there was no effect of stress. We conclude that chronic fluoxetine treatment induced a decreased intake of sweet food, as well as a reduction in leptin levels, and that this result could represent a compensatory response to reduced food intake rather than a direct anorectic mechanism. No interaction with chronic stress was observed. PMID- 18423830 TI - The effect of lamotrigine on platelet monoamine oxidase type B activity in patients with bipolar depression. AB - Lamotrigine is an anticonvulsant drug effective in the treatment of epilepsy and bipolar depression. Preclinical data showed that lamotrigine inhibited monoamine oxidase (MAO) activity in vitro. The aim of the study was to determine the effects of 6-weeks lamotrigine treatment on platelet MAO type B (MAO-B) activity in patient with bipolar depression. The study included 26 female patients with bipolar I disorder in depressive episode (DSM-IV criteria, Hamilton Depression Rating Scale (HAMD) and Young Mania Rating Scale). Platelet MAO-B activity was determined spectrofluorimetrically before and after 6 weeks of the treatment with a relatively low dose of lamotrigine (100 mg/day). Six weeks of treatment with lamotrigine significantly decreased platelet MAO-B activity in bipolar depressed patients. This inhibitory effect was not related to smoking status and was independent of the treatment combinations (lamotrigine alone or in combination with either lithium or antipsychotics). Lamotrigine treatment induced a decrease in total HAMD scores in bipolar depressed patients, which was not significantly correlated with reduction of platelet MAO-B activity. These findings provide in vivo insight of lamotrigine effect on platelet MAO-B activity in patients with bipolar depression. Its in vivo MAO-B inhibiting effect might have contributed in part to its antidepressant activity. PMID- 18423831 TI - T-cell regulation in chronic paranasal sinus disease. AB - BACKGROUND: Chronic rhinosinusitis is an inflammatory disease with distinct cytokine and remodeling patterns. Chronic rhinosinusitis with nasal polyps (CRSwNP) is characterized by a T(H)2-skewed eosinophilic inflammation, whereas chronic rhinosinusitis without nasal polyps (CRSsNP) represents a predominant T(H)1 milieu. OBJECTIVE: We aimed to study the direct tissue expression of transcription factors for T-cell subpopulations, including T regulatory cells, in relation to the cytokine expression patterns in the different disease subgroups. METHODS: The expression of forkhead box P3 (FOXP3), T-box transcription factor (T bet), GATA-3, retinoid acid-related orphan receptor C (RORc), the suppressive cytokines TGF-beta1 and IL-10, and T(H)1/ T(H)2/ T(H)17 cytokines (IFN-gamma, IL 4, IL-5, IL-13, IL-17) were analyzed by means of RT-PCR in 13 CRSsNP, 16 CRSwNP, and 10 control samples. Additional protein measurements were performed for TGF beta1 and IFN-gamma. RESULTS: In CRSwNP, we observed a significantly lower FOXP3 mRNA and TGF-beta1 protein expression, but a significantly higher T-bet, GATA-3, IL-5, and IL-13 mRNA expression compared with controls, whereas RORc was not significantly different compared with controls. In CRSsNP, FOXP3, T-bet, GATA-3, and RORc expression was not significantly different from controls, whereas TGF beta1 mRNA, IFN-gamma mRNA, and protein were significantly higher in CRSsNP compared with controls. For IL-17, no significant differences were noted among all groups. CONCLUSION: We demonstrate for the first time a decreased FOXP3 expression accompanied by an upregulation of T-bet and GATA-3 and a downregulation of TGF-beta1 in CRSwNP versus controls and CRSsNP. PMID- 18423832 TI - Highly integrated single-base resolution maps of the epigenome in Arabidopsis. AB - Deciphering the multiple layers of epigenetic regulation that control transcription is critical to understanding how plants develop and respond to their environment. Using sequencing-by-synthesis technology we directly sequenced the cytosine methylome (methylC-seq), transcriptome (mRNA-seq), and small RNA transcriptome (smRNA-seq) to generate highly integrated epigenome maps for wild type Arabidopsis thaliana and mutants defective in DNA methyltransferase or demethylase activity. At single-base resolution we discovered extensive, previously undetected DNA methylation, identified the context and level of methylation at each site, and observed local sequence effects upon methylation state. Deep sequencing of smRNAs revealed a direct relationship between the location of smRNAs and DNA methylation, perturbation of smRNA biogenesis upon loss of CpG DNA methylation, and a tendency for smRNAs to direct strand-specific DNA methylation in regions of RNA-DNA homology. Finally, strand-specific mRNA-seq revealed altered transcript abundance of hundreds of genes, transposons, and unannotated intergenic transcripts upon modification of the DNA methylation state. PMID- 18423835 TI - Involvement of cyclin-dependent kinase 5 in 2,5-hexanedione-induced neuropathy. AB - Occupational exposure to n-hexane produces a neuropathy characterized as a central-peripheral distal axonopathy, which is mediated by 2,5-hexanedione (HD). To investigate the mechanisms of the neuropathy induced by HD, the contents and activities of cyclin-dependent kinase 5 (CDK5) and activators (p35 precursor, p35 and p25) in rats' cerebrum cortex (CC), spinal cord (SC) and sciatic nerve (SN) were determined. The results showed that the levels and activities of CDK5 in CC of 200 or 400mg/kg HD-treated rats were significantly decreased in both the cytosolic and membrane fractions and negatively correlated with gait abnormality in the cytosolic fraction. However, CDK5 contents and activities in SN of rats treated with 200 or 400mg/kg HD were significantly increased and positively correlated with gait abnormality in both the cytosolic and membrane fractions. Although increases of CDK5 contents in both the cytosolic and membrane fractions of SC in 200 and 400mg/kg HD-treated rats were also observed, CDK5 activities were significantly decreased in the cytosolic fraction and negatively correlated with gait abnormality. The changes of p35 precursor, p35 and p25 contents in CC, SC and SN showed the same pattern with that of CDK5 activities. Thus, HD intoxication was associated with deregulation of CDK5 and its activator p35 or p25 in nerve tissues. The inconsistent changes of CDK5 activities in CNS and PNS might delegate the different mechanisms of HD-induced peripheral neuropathy. PMID- 18423833 TI - Alteration of dopamine uptake into rat striatal vesicles and synaptosomes caused by an in vitro exposure to atrazine and some of its metabolites. AB - Studies have shown that both in vivo and in vitro exposure to the herbicide atrazine (ATR) results in dopaminergic neurotoxicity manifested by decreased striatal dopamine (DA) levels. However, the mechanism behind this reduction is largely unknown. A decrease in striatal DA could be due to ATR exposure affecting vesicular and/or synaptosomal uptake resulting in disrupted vesicular storage and/or cellular uptake of DA. Hence, we investigated the effects of in vitro ATR exposure on DA uptake into isolated rat striatal synaptosomes and synaptic vesicles. In addition to ATR, effects of its major mammalian metabolites, didealkyl atrazine (DACT), desethyl atrazine (DE) and desiopropyl atrazine (DIP) were investigated. ATR (1-250 microM) inhibited DA uptake into synaptic vesicles in a dose-dependent manner. Of the three ATR metabolites tested, DACT did not affect vesicular DA uptake. DE and DIP, on the other hand, significantly decreased vesicular DA uptake with the effect of 100 microM DE/DIP being similar to the effect of the same concentration of ATR. Kinetic analysis of vesicular DA uptake indicated that ATR significantly decreased the V(max) while the K(m) value was not affected. Contrary to the inhibitory effects on vesicular DA uptake, synaptosomal DA uptake was marginally (6-13%) increased by ATR and DE, but not by DACT and DIP, at concentrations of or=500 mg/kg/day (3,000 mg/m(2)/day) induced mortality. In a 90-day toxicity study in rats, no gross toxicity was seen at doses of 30, 100, or 300 mg/kg/day (180, 600, or 1,800 mg/m(2)/day, respectively). Antalarmin (300 mg/kg/day) induced mild anemia, increases in serum gamma-glutamyl transferase activity, and microscopic hepatic pathology (bile duct hyperplasia and epithelial necrosis, periportal inflammation). Microscopic renal changes (cortical necrosis, inflammation, hypertrophy, nephropathy) were observed in rats at all Antalarmin doses. In a 14 day range-finding study in dogs, Antalarmin doses >or=50mg/kg/day (1,000 mg/m(2)/day) induced repeated emesis and bone marrow suppression. In a 90-day toxicity study in dogs, Antalarmin (4, 8, or 16 mg/kg/day (80, 160, or 320 mg/m(2)/day, respectively)) induced bone marrow and lymphoid depletion, but no gross toxicity. Comparative in vitro studies using rat, dog, and human neutrophil progenitors demonstrated that canine bone marrow cells are highly sensitive to Antalarmin cytotoxicity, while rat and human bone marrow cells are relatively insensitive. As such, the bone marrow toxicity observed in dogs is considered likely to over-predict Antalarmin toxicity in humans. The hepatic and renal toxicities seen in rats exposed to Antalarmin identify those tissues as the most likely targets for Antalarmin toxicity in humans. PMID- 18423836 TI - Effect of serum starvation and chemical inhibitors on cell cycle synchronization of canine dermal fibroblasts. AB - The cell cycle stage of donor cells and the method of cell cycle synchronization are important factors influencing the success of somatic cell nuclear transfer. In this study, we examined the effects of serum starvation, culture to confluence, and treatment with chemical inhibitors (roscovitine, aphidicolin, and colchicine) on cell cycle characteristics of canine dermal fibroblast cells. The effect of the various methods of cell cycle synchronization was determined by flow cytometry. Short periods of serum starvation (24-72 h) increased (P<0.05) the proportion of cells at the G0/G1 phase (88.4-90.9%) as compared to the control group (73.6%). A similar increase in the percentage of G0/G1 (P<0.05) cells were obtained in the culture to confluency group (91.8%). Treatment with various concentrations of roscovitine did not increase the proportion of G0/G1 cells; conversely, at concentrations of 30 and 45 microM, it increased (P<0.05) the percentage of cells that underwent apoptosis. The use of aphidicolin led to increase percentages of cells at the S phase in a dose-dependent manner, without increasing apoptosis. Colchicine, at a concentration of 0.1 microg/mL, increased the proportion of cells at the G2/M phase (38.5%, P<0.05); conversely, it decreased the proportions of G0/G1 cells (51.4%, P<0.05). Concentrations of colchicines >0.1 microg/mL did not increase the percentage of G2/M phase cells. The effects of chemical inhibitors were fully reversible; their removal led to a rapid progression in the cell cycle. In conclusion, canine dermal fibroblasts were effectively synchronized at various stages of the cell cycle, which could have benefits for somatic cell nuclear transfer in this species. PMID- 18423838 TI - Synergistic effects of the dopaminergic and glutamatergic system on hippocampal volume in alcohol-dependent patients. AB - Several genes of the dopaminergic and glutamatergic neurotransmitter systems have been found to be associated with alcohol disease and related intermediate phenotypes. Here, we evaluated genetic variants of the catechol-O methyltransferase (COMT) and the metabotropic glutamate receptor 3 (mGluR3) genes in alcohol-dependent patients and their association with volumetric measurements of brain structures. By combined analysis of imaging data and genotyping results, large numbers of variables are produced that overstrain conventional statistical methods based on tests for group differences. Limitations in assessment of epistatic effects and multiple testing problems are encountered. Therefore, we introduce a novel method for detecting associations between a set of genetic markers and phenotypical measurements based on machine learning techniques. Hippocampal volume was found to be associated with epistatic effects of the COMT mGluR3 genes in alcohol-dependent patients but not in controls. These data are in line with prior studies supporting a role for dopamine-glutamate interaction in modulation of alcohol disease. PMID- 18423837 TI - Quantifying the heritability of task-related brain activation and performance during the N-back working memory task: a twin fMRI study. AB - Working memory-related brain activation has been widely studied, and impaired activation patterns have been reported for several psychiatric disorders. We investigated whether variation in N-back working memory brain activation is genetically influenced in 60 pairs of twins, (29 monozygotic (MZ), 31 dizygotic (DZ); mean age 24.4+/-1.7S.D.). Task-related brain response (BOLD percent signal difference of 2 minus 0-back) was measured in three regions of interest. Although statistical power was low due to the small sample size, for middle frontal gyrus, angular gyrus, and supramarginal gyrus, the MZ correlations were, in general, approximately twice those of the DZ pairs, with non-significant heritability estimates (14-30%) in the low-moderate range. Task performance was strongly influenced by genes (57-73%) and highly correlated with cognitive ability (0.44 0.55). This study, which will be expanded over the next 3 years, provides the first support that individual variation in working memory-related brain activation is to some extent influenced by genes. PMID- 18423839 TI - Effect of air-drying and solvent evaporation on the strength of HEMA-rich versus HEMA-free one-step adhesives. AB - OBJECTIVES: The objectives of this study were (1) to clarify the relationship between the duration of air-drying of one-step self-etch adhesives (1-SEAs) and the evaporation degree (ED) of solvents, and (2) to evaluate the effect of ED on the ultimate micro-tensile strength (microTS) of the adhesives. METHODS: The ED of one HEMA-rich 1-SEA, Clearfil S3 Bond (Kuraray), and two HEMA-free 1-SEAs, iBond (Heraeus-Kulzer) and G-Bond (GC), was determined without air-drying and after, respectively, 5- and 10-s air-drying using a gravimetric method. Next, the microTS of the adhesives at ED equivalent to 0-, 5- and 10-s air-drying was measured. RESULTS: The ED increased with extension of air-drying time. Among the adhesives tested, iBond showed the largest ED, followed by G-Bond and Clearfil S3 Bond in this order. A longer air-drying time for 10-s resulted in a statistically significantly higher microTS for the HEMA-rich Clearfil S3 Bond. The microTS of the latter was higher than that of the other two HEMA-free adhesives for each air drying time. SIGNIFICANCE: Air-drying of 1-SEAs had a significant effect on the degree of solvent evaporation (ED) and also on the mechanical properties (microTS) of the 1-SEAs upon setting. It is therefore beneficial to remove solvents of the 1-SEAs as much as possible by thorough, strong air-drying in order to achieve a strong adhesive layer at the interface. PMID- 18423840 TI - Application of analytical stress solutions to bi-axially loaded dental ceramic dental cement bilayers. AB - OBJECTIVE: Increased consideration of dental ceramics and dental cements as a singular structural unit for in-vitro mechanical testing has resulted in the reporting of a wide range of analytical methods to calculate the failure stresses. Therefore a comparison of observations between studies is complicated by the use of dissimilar stress solutions despite the employment of a similar testing methodology and specimen geometry. MATERIALS AND METHODS: Three analytical solutions to calculate failure stresses in bi-axially loaded dental ceramic-dental cement bilayers were appraised for a commonly utilized testing geometry (ball-on-ring). Clinically relevant datasets were generated from the bi axial flexure testing of uncoated and dental cement coated aluminous core ceramic exposed to differing ceramic surface preparations. A Weibull statistical approach was utilized in order to provide insight into the impact of the analytical method on both the scale (sigma0) and distribution (m) of the failure stress data. RESULTS: Calculation of the bi-axial flexure stress utilizing Timoshenko's analysis resulted in an increase in sigma0 for the uncoated (6%) and dental cement coated (11-12%) aluminous core ceramic, when compared with the bilayered solutions reported by Rosenstiel and Hsueh. However, the shape of the failure distributions illustrated by the consistency of m and associated 95% confidence intervals was not influenced by the analytical stress solution employed. SIGNIFICANCE: The choice of the analytical method chosen to calculate failure stresses in bi-axially loaded dental ceramic-dental cement bilayers will impact on the magnitude of the reported strength. Comparison between the failure stresses of uncoated and cement coated dental ceramics is more accurately represented by bilayer solutions, which account for the mismatch between the elastic constants of dissimilar materials. However, within the context of dental cement coated dental ceramics of clinically relevant dimensions, the choice of solution is unlikely to impact on the interpretation of the observations previously reported in the dental literature. PMID- 18423841 TI - Environmental service payments: evaluating biodiversity conservation trade-offs and cost-efficiency in the Osa Conservation Area, Costa Rica. AB - The cost-efficiency of payments for environmental services (PES) to private landowners in the Osa Conservation Area, Costa Rica, is evaluated in terms of the trade-off between biodiversity representation and opportunity costs of conservation to agricultural and forestry land-use. Using available GIS data and an 'off-the-shelf' software application called TARGET, we find that the PES allocation criteria applied by authorities in 2002-2003 were more than twice as cost-efficient as criteria applied during 1999-2001. Results show that a policy relevant assessment of the cost-effectiveness of PES relative to other conservation policies can be carried out at regional level using available studies and GIS data. However, there are a number of data and conceptual limitations to using heuristic optimisation algorithms in the analysis of the cost-efficiency of PES. Site specific data on probabilities of land-use change, and a detailed specification of opportunity costs of farm land, labour and capital are required to use algorithms such as TARGET for ranking individual sites based on cost-efficiency. Despite its conceptual soundness for regional conservation analysis, biodiversity complementarity presents a practical challenge as a criterion for PES eligibility at farm level because it varies depending on the set of areas under PES contracts at any one time. PMID- 18423842 TI - Aerospace wetland monitoring by hyperspectral imaging sensors: a case study in the coastal zone of San Rossore Natural Park. AB - The San Rossore Natural Park, located on the Tuscany (Italy) coast, has been utilized over the last 10 years for many remote sensing campaigns devoted to coastal zone monitoring. A wet area is located in the south-west part of the Natural Park and it is characterized by a system of ponds and dunes formed by sediment deposition occurring at the Arno River estuary. The considerable amount of collected data has permitted us to investigate the evolution of wetland spreading and land coverage as well as to retrieve relevant biogeochemical parameters, e.g. green biomass, from remote sensing images and products. This analysis has proved that the monitoring of coastal wetlands, characterized by shallow waters, moor and dunes, demands dedicated aerospace sensors with high spatial and spectral resolution. The outcomes of the processing of images gathered during several remote sensing campaigns by airborne and spaceborne hyperspectral sensors are presented and discussed. A particular effort has been devoted to sensor response calibration and data validation due to the complex heterogeneity of the observed natural surfaces. PMID- 18423843 TI - The management of household hazardous waste in the United Kingdom. AB - Waste legislation in the United Kingdom (UK) implements European Union (EU) Directives and Regulations. However, the term used to refer to hazardous waste generated in household or municipal situations, household hazardous waste (HHW), does not occur in UK, or EU, legislation. The EU's Hazardous Waste Directive and European Waste Catalogue are the principal legislation influencing HHW, although the waste categories described are difficult to interpret. Other legislation also have impacts on HHW definition and disposal, some of which will alter current HHW disposal practices, leading to a variety of potential consequences. This paper discusses the issues affecting the management of HHW in the UK, including the apparent absence of a HHW-specific regulatory structure. Policy and regulatory measures that influence HHW management before disposal and after disposal are considered, with particular emphasis placed on disposal to landfill. PMID- 18423844 TI - Remote sensing water observation for supporting Lake Victoria weed management. AB - This paper aims to assess the suitability of remote sensing for enhancing the management of water body resources and for providing an inexpensive way to gather, on a wide area, weed infestation extent and optical parameter linked to the water body status. Remotely sensed satellite images and ancillary ground true data were used to produce land cover maps, trough classification techniques, and water compounds maps, applying radiative transfer models. The study proposed within the framework of the cooperation between Italian Foreign Affair Ministry (through the University of Rome) and Kenyan Authorities has been carried out on the Kenyan part of the Lake Victoria. This lake is one of the largest freshwater bodies of the world where, over the last few years environmental challenges and human impact have perturbed the ecological balance affecting the biodiversity. The objective of this research study is to define the thematic products, retrievable from satellite images, like weed abundance maps and water compound concentrations. These products, if provided with an appropriate time frequency, are useful to identify the preconditions for the occurrence of hazard events like abnormal macrophyte proliferation and to develop an up-to-date decision support system devoted to an apprised territory, environment and resource management. PMID- 18423845 TI - Application of remote and in situ information to the management of wetlands in Poland. AB - The protection and regeneration of wetlands has been of crucial importance as a goal in ecological research and in nature conservation for some time and is more important than ever now. Knowledge about the biophysical properties of wetlands' vegetation retrieved from satellite images enables us to improve the monitoring of these unique areas, which are otherwise very often impenetrable and therefore difficult to examine, analyze and assess by means of site visits. The Biebrza Wetlands are situated in the North-East part of Poland and are one of the largest areas made up of marshes and swamps in the entire EU. This is still one of the wildest areas and one of the least destroyed, damaged or changed by human impact. However, in the recent decades there have been attempts made to intensify and overexploit the natural resources of the region and implement new agriculture practices in the area. In this period, drainage canals have been built, and a good deal of the area has been drained. The area of this precious ecosystem covers 25 494 ha. This valuable area of peat with unique vegetation species and with very special birds is one of the most valuable areas in Europe and in 1995 was added to the list of Ramsar sites. The investigation of wetlands in the Biebrza River Valley has been carried out at ground level by taking measurements of soil moisture, evapotranspiration, Leaf Area Index, wet and dry biomass and the levels of ground water and meteorological parameters. Also examined were radiative temperature, detailed vegetation mapping, and APAR. For some years the deterioration of peat lands has been noticed due to the drying out of the area and the frequent outbreak of fires. The consequence is the succession of new vegetation and the appearance of new ecosystems. The Remote Sensing Centre in the Institute of Geodesy and Cartography has undertaken the investigation by applying ERS-2.SAR and ENVISAT ASAR of IS2 and IS4 and VV, HH, HV polarization for the purpose of modeling soil moisture and humidity changes of the area under investigation. The investigation also aimed at finding the best biophysical properties of wetlands' vegetation to characterize marshland habitats and its changes. At the same time as registering the microwave data, the optical data from Landsat ETM+, SPOT VEGETATION, ERS-2.ATSR, ENVISAT MERIS, and NOAA/AVHRR have been registered and information about the biomass and heat fluxes as sensible and latent heat has also been calculated. The vegetation indices are calculated from EO satellite data taking into account jointly the features of vegetation responsible for reflection in various bands and combining this information from several spectral bands. Also, the changes in the humidity of the area have been examined by extracting the backscattering coefficients from two SAR images that were taken at a similar period of the year but with a gap of 5 years. The information about soil moisture as retention, soil moisture changes, heat fluxes and evapotranspiration are all very important for estimates of CO(2) sequestration. The ENVISAT images have been obtained for the ESA AO-ID122 project. Also the SMOS and ALOS data will be applied for the Biebrza Wetlands in the future. PMID- 18423846 TI - The purified recombinant precursor of rat mitochondrial dimethylglycine dehydrogenase binds FAD via an autocatalytic reaction. AB - The precursor of the rat mitochondrial flavoenzyme dimethylglycine dehydrogenase (Me(2)GlyDH) has been produced in Escherichia coli as a C-terminally 6-His-tagged fusion protein, purified by one-step affinity chromatography and identified by ESI-MS/MS. It was correctly processed into its mature form upon incubation with solubilized rat liver mitoplasts. The purified precursor was mainly in its apo form as demonstrated by immunological and fluorimetric detection of covalently bound flavin adenine dinucleotide (FAD). Results described here definitively demonstrate that: (i) covalent attachment of FAD to Me(2)GlyDH apoenzyme can proceed in vitro autocatalytically, without third reactants; (ii) the removal of mitochondrial presequence by mitochondrial processing peptidase is not required for covalent autoflavinylation. PMID- 18423847 TI - Maintenance and characterization of spontaneous contraction rhythm in cultured cardiac myocytes fused with cardiac fibroblasts. AB - Cardiomyocytes (CMs) fuse with various cells including endothelial cells, cardiac fibroblasts (CFs). In addition, recent studies have shown that stem cells fuse spontaneously with cells remaining in the damaged tissues, and restore tissue functions after myocardial infarction. In this study, we investigated whether cultured cardiomyocytes fused with proliferative cardiac fibroblasts maintained the phenotype of functional myocytes by analyzing the spontaneous contraction rhythm after fusion with CFs lacking a beating capability. CMs and CFs cultured for 4 days in vitro were used in this study. The fusion of cultured CMs and CFs was achieved with polyethylene glycol (PEG) and hemagglutinating virus of Japan (HVJ). Analyses of CMs fused with CFs by using either PEG or HVJ to imitate spontaneous fusion in vivo demonstrated that CMs and CFs actually fused together and fused cells expressed lineage marker proteins of both CMs and CFs. In addition, fused cells reentered the G2-M phase of the cell cycle. Furthermore, fused cells retained the spontaneous contraction activity. The present study demonstrated that CMs fused with proliferative CFs showed the phenotype of both CMs and CFs and spontaneous rhythmic contraction. PMID- 18423848 TI - Cholecystokinin (CCK) and CCK receptor expression by human gliomas: Evidence for an autocrine/paracrine stimulatory loop. AB - Cholecystokinin (CCK) is a gut-brain peptide has been described to be able to induce mitosis according to recent studies. Additionally, conflicting data has been published on whether tumours of the central and peripheral nervous system in general, and gliomas in particular, express CCK receptors. In the present in vitro study we employed reverse transcription followed by the polymerase chain reaction (RT-PCR) to investigate whether mRNA for CCK-A and CCK-B receptors as well as CCK peptide itself is present in primary human gliomas and the U-87 MG GBM cell line. The data show that 14/14 (100%) of the primary gliomas exhibited mRNA expression for the CCK peptide gene and the B receptor including the U-87 MG cells, whereas, only 2/14 (14%) showed presence of the CCK-A receptor. The presence of CCK receptors together with CCK peptide expression itself suggests presence of an autocrine loop controlling glioma cell growth. In support of this conclusion, a neutralizing antibody against the CCK peptide exhibited a dose dependent inhibition of cell growth whereas, antagonists to CCK caused a dose depend inhibition of exogenous stimulated glioma cell growth in vitro, via the CCK-B receptor which is PKC activated. Assessment of apoptosis and proteasome activity were undertaken and we report that treatment with CCK antagonists decreased proteasome and increased caspase-3 activity. These data indicate that CCK peptide and CCK-B are abundant in human gliomas and they act to stimulate cell growth in an autocrine manner, primarily via the high affinity CCK-B receptor, which was blocked by antagonists to CCK, perhaps via apoptosis. PMID- 18423849 TI - (Neuro-)endocrinology of epithelial hair follicle stem cells. AB - The hair follicle is a repository of different types of somatic stem cells. However, even though the hair follicle is both a prominent target organ and a potent, non-classical site of production and/or metabolism of numerous polypetide and steroid hormones, neuropeptides, neurotransmitters and neurotrophins, the (neuro-)endocrine controls of hair follicle epithelial stem cell (HFeSC) biology remain to be systematically explored. Focussing on HFeSCs, we attempt here to offer a "roadmap through terra incognita" by listing key open questions, by exploring endocrinologically relevant HFeSC gene profiling and mouse genomics data, and by sketching several clinically relevant pathways via which systemic and/or locally generated (neuro-)endocrine signals might impact on HFeSC. Exemplarily, we discuss, e.g. the potential roles of glucocorticoid and vitamin D receptors, the hairless gene product, thymic hormones, bone morphogenic proteins (BMPs) and their antagonists, and Skg-3 in HFeSC biology. Furthermore, we elaborate on the potential role of nerve growth factor (NGF) and substance P dependent neurogenic inflammation in HFeSC damage, and explore how neuroendocrine signals may influence the balance between maintenance and destruction of hair follicle immune privilege, which protects these stem cells and their progeny. These considerations call for a concerted research effort to dissect the (neuro )endocrinology of HFeSCs much more systematically than before. PMID- 18423850 TI - International Working Group on MDS cytogenetics: October 2007 meeting report. AB - The inaugural meeting of the International Working Group on MDS cytogenetics convened 22-23 October 2007 in Chicago, IL. Under the sponsorship of the Myelodysplastic Syndromes Foundation, the group was organized to address the substantial need for worldwide standardized cytogenetic testing for MDS in clinical practice and research. Eighteen cytogeneticists from 10 countries attended the first working group meeting. Representatives from France and Austria were unable to attend the Chicago meeting. Marilyn L. Slovak, PhD (City of Hope, USA) served as Working Group Chair and Gordon Dewald, PhD (Mayo Clinic, USA), served as Working Group Advisor and Co-Chair. Other members in attendance included: Mette Andersen, Rigshospitalet, Denmark; Lynda Campbell, St. Vincent's Hospital Melbourne, Australia; Athena Cherry, Stanford University, USA; Kathy Chun, North York General Hospital, Canada; Mike Griffiths, West Midlands Regional Genetics Lab, UK; Detlef Haase, Georg-August-Universitat, Germany; Claudia Haferlach, MLL Munchner Leukamielabor GmbH, Germany; Anne Hagemeijer, University of Leuven, Belgium; Barbara Hildebrandt, Institut fur Humangenetik & Anthropologie Dupsilonsseldorf, Germany; Douglas Horsman, BC Cancer Agency, Canada; M. Anwar Iqbal, University of Rochester, USA; Suresh Jhanwar, Memorial Sloan-Kettering Cancer Center, USA; Bertil Johansson, University Hospital, Sweden; Michelle LeBeau, University of Chicago, USA; Kazuma Ohyashiki, Tokyo Medical University, Japan; Francesc Sole, Hospital del Mar, Spain. The focus of the working group was to establish the natural history and clinical significance of cytogenetic anomalies associated with the myelodysplastic syndromes (MDS), and to incorporate cytogenetic testing into the development of new treatments to cure MDS. Three specific goals were discussed in an effort to rapidly improve the care of patients with MDS. The first goal was how to educate physicians on the appropriate use of cost effective cytogenetic testing for their patients with MDS. The second goal discussed was how best this working group could assist pharmaceutical companies with the use of appropriate cytogenetic testing in their evaluation of new drugs. The final goal discussed was how to advance cytogenetic research into the origin, progression and clinical significance of genetic anomalies associated with MDS. PMID- 18423851 TI - The benefit of image guidance for the contralateral interhemispheric approach to the lateral ventricle. AB - OBJECTIVES: Recently, neurosurgeons have increasingly faced small intracerebral lesions in asymptomatic or minimally symptomatic patients. Here, we evaluated a series of four patients with nearly asymptomatic intraventricular tumors close to the corpus callosum that had been treated with the aid of an image-guided transcallosal approach. PATIENTS AND METHODS: Four consecutive patients suffering from left intra- and paraventricular tumors were operated on via a contralateral interhemispheric transcallosal approach with the aid of neuronavigation. Our image-guided system directed: (1) the skin incision, (2) the interhemispheric dissection, and (3) the incision of the corpus callosum. RESULTS: Using the image guided contralateral interhemispheric transcallosal approach to the left ventricle all lesions have been completely resected without the risk of damage to the dominant hemisphere. The callosal incision was kept as limited as possible (1.2-2.1cm) depending on the size of the tumor. No postoperative neurological or neuropsychological deficit was observed in our series. CONCLUSION: Neuronavigation facilitates a safe and targeted contralateral interhemispheric transcallosal approach to the dominant hemisphere's lateral ventricle. Our technique minimizes the risk of damage to the dominant hemisphere and requires only a limited opening of the corpus callosum, which might decrease the risk of neuropsychological morbidity. PMID- 18423852 TI - Decolorization of direct dyes by immobilized turnip peroxidase in batch and continuous processes. AB - An inexpensive immobilized turnip peroxidase has been employed for the decolorization of some direct dyes in batch and continuous reactors. Wood shaving was investigated as an inexpensive material for the preparation of bioaffinity support. Concanavalin A-wood shaving bound turnip peroxidase exhibited 67% of the original enzyme activity. Both soluble and immobilized turnip peroxidase could effectively remove more than 50% color from dyes in the presence of metals/salt and 0.6mM 1-hydroxybenzotriazole, after 1h of incubation. The columns containing immobilized peroxidase could decolorize 64% direct red 23% and 50% mixture of direct dyes at 4 and 3 months of operation, respectively. Total organic carbon analysis of treated dye or mixture of dyes revealed that these results were quite comparable to the loss of color from solutions. Thus, this study showed that the immobilized enzyme could be efficiently used for the removal of synthetic dyes from industrial effluents. PMID- 18423853 TI - Evaluation of a municipal landfill site in Southern Spain with GIS-aided methodology. AB - Landfill siting should take into account a wide range of territorial and legal factors in order to reduce negative impacts on the environment. This article describes a landfill siting method, which is based on EVIAVE, a landfill diagnosis method developed at the University of Granada. Geographical Information Systems (GIS) technology is also used to generate spatial data for site assessment. Landfill site suitability is assessed on a scale based on territorial indices that measure the risk of contamination for the following five environmental components: surface water, groundwater, atmosphere, soil, and human health. The method described in this article has been used to evaluate an area in Granada (Spain) where there is a currently operating landfill. The results obtained show that suitable locations for the disposal of municipal waste were successfully identified. The low environmental index values reflect the suitability of this landfill site as well as its minimal negative impacts on the environment. PMID- 18423854 TI - Sorption behavior of tetrabromobisphenol A in two soils with different characteristics. AB - Sorption of tetrabromobisphenol A (TBBPA) in soil influences its fate and transport in the environment. The sorption behaviors of TBBPA in two soils with different characteristics were investigated using batch equilibration experiments in the study, and the impacts of ionic strength and pH on the sorption were also evaluated. The results showed that the fast sorption rather than the slow sorption played a main role in the sorption process. The nonlinear sorption isotherms of TBBPA in the soils were observed and the Freundlich model could describe the sorption behavior of TBBPA well. The calculated KF were 78.5 and 364.6 (mg/kg)(mg/L)-n for LN soil (loamy clay) and GX soil (silt loam), respectively. Soil organic matter (SOM) played a main role in the sorption of TBBPA, which contributed about 90% to the sorption in the soils. The desorption hysteresis of TBBPA was revealed in the single-cycle sorption and desorption experiments. The sorbed amount of TBBPA decreased with the increase in solution pH and increased with the increase in ionic strength. It was found that the effects of ionic strength on the sorption were mainly caused by the change of solution pH. PMID- 18423855 TI - Environmental liability and life-cycle management of used lubricating oils. AB - Used oil handling, as a business, requires an extensive understanding by management that environmental liabilities exist through its supply chain. Findings from a review of the legal requirements of operating a used oil handling business were: understanding the transfer of ownership of used petroleum hydrocarbons is critical to any such business and how this is documented; used oil handlers are responsible for providing training to their staff, including site personnel and any third party waste contractors, and for communicating best practice procedures relating to the management of used petroleum hydrocarbons to all those individuals and organisations involved in business relationships that the used oil handling companies have; used oil handlers should audit the performance of any third party contractors that it engages to conduct work on behalf of its customers. Hypothetical situations of a company planning to enter the used oil handling market are described in relation to petroleum hydrocarbon wastes it handles to illustrate the range of potential liabilities. Companies proposing to establish a used oil handling business should ensure that they provide accurate advice to its employees, its customer's employees and to its third party contractors, all of which may be responsible for handling used petroleum hydrocarbons as part of the service it intends to provide, and that it has a well documented system addressing how environmental issues are managed. PMID- 18423856 TI - Decontamination of soil washing wastewater using solar driven advanced oxidation processes. AB - Decontamination of soil washing wastewater was performed using two different solar driven advanced oxidation processes (AOPs): the photo-Fenton reaction and the cobalt/peroxymonosulfate/ultraviolet (Co/PMS/UV) process. Complete sodium dodecyl sulphate (SDS), the surfactant agent used to enhance soil washing process, degradation was achieved when the Co/PMS/UV process was used. In the case of photo-Fenton reaction, almost complete SDS degradation was achieved after the use of almost four times the actual energy amount required by the Co/PMS/UV process. Initial reaction rate in the first 15min (IR15) was determined for each process in order to compare them. Highest IR15 value was determined for the Co/PMS/UV process (0.011mmol/min) followed by the photo-Fenton reaction (0.0072mmol/min) and the dark Co/PMS and Fenton processes (IR15=0.002mmol/min in both cases). Organic matter depletion in the wastewater, as the sum of surfactant and total petroleum hydrocarbons present (measured as chemical oxygen demand, COD), was also determined for both solar driven processes. It was found that, for the case of COD, the highest removal (69%) was achieved when photo-Fenton reaction was used whereas Co/PMS/UV process yielded a slightly lower removal (51%). In both cases, organic matter removal achieved was over 50%, which can be consider proper for the coupling of the tested AOPs with conventional wastewater treatment processes such as biodegradation. PMID- 18423857 TI - Reducing aluminum dust explosion hazards: case study of dust inerting in an aluminum buffing operation. AB - Metal powders or dusts can represent significant dust explosion hazards in industry, due to their relatively low ignition energy and high explosivity. The hazard is well known in industries that produce or use aluminum powders, but is sometimes not recognized by facilities that produce aluminum dust as a byproduct of bulk aluminum processing. As demonstrated by the 2003 dust explosion at aluminum wheel manufacturer Hayes Lemmerz, facilities that process bulk metals are at risk due to dust generated during machining and finishing operations [U.S. Chemical Safety and Hazard Investigation Board, Investigation Report, Aluminum Dust Explosion Hayes Lemmerz International, Inc., Huntington, Indiana, Report No. 2004-01-I-IN, September 2005]. Previous studies have shown that aluminum dust explosions are more difficult to suppress with flame retardants or inerting agents than dust explosions fueled by other materials such as coal [A.G. Dastidar, P.R. Amyotte, J. Going, K. Chatrathi, Flammability limits of dust minimum inerting concentrations, Proc. Saf. Progr., 18-1 (1999) 56-63]. In this paper, an inerting method is discussed to reduce the dust explosion hazard of residue created in an aluminum buffing operation as the residue is generated. This technique reduces the dust explosion hazard throughout the buffing process and within the dust collector systems making the process inherently safer. Dust explosion testing results are presented for process dusts produced during trials with varying amounts of flame retardant additives. PMID- 18423858 TI - Reductive dechlorination of 3,3',4,4'-tetrachlorobiphenyl (PCB77) using palladium or palladium/iron nanoparticles and assessment of the reduction in toxic potency in vascular endothelial cells. AB - Palladium-based nanoparticles immobilized in polymeric matrices were applied to the reductive dechlorination of 3,3',4,4'-tetrachlorobiphenyl (PCB77) at room temperature. Two different dechlorination platforms were evaluated using (1) Pd nanoparticles within conductive polypyrrole films; or (2) immobilized Fe/Pd nanoparticles within polyvinylidene fluoride microfiltration membranes. For the first approach, the polypyrrole film was electrochemically formed in the presence of perchlorate ions that were incorporated into the film to counter-balance the positive charges of the polypyrrole chain. The film was then incubated in a solution containing tetrachloropalladate ions, which were exchanged with the perchlorate ions within the film. During this exchange, reduction of tetrachloropalladate by polypyrrole occurred, which led to the formation of palladium nanoparticles within the film. For the second approach, the membrane supported Fe/Pd nanoparticles were prepared in three steps: polymerization of acrylic acid in polyvinylidene fluoride microfiltration membrane pores was followed by ion exchange of Fe(2+), and then chemical reduction of the ferrous ions bound to the carboxylate groups. The membrane-supported iron nanoparticles were then soaked in a solution of tetrachloropalladate resulting in the deposition of Pd on the Fe surface. The nanoparticles prepared by both approaches were employed in the dechlorination of PCB77. The presence of hydrogen was required when the monometallic Pd nanoparticles were employed. The results indicate the removal of chlorine atoms from PCB77, which led to the formation of lower chlorinated intermediates and ultimately biphenyl. Toxicity associated with vascular dysfunction by PCB77 and biphenyl was compared using cultured endothelial cells. The data strongly suggest that the dechlorination system used in this study markedly reduced the proinflammatory activity of PCB77, a persistent organic pollutant. PMID- 18423859 TI - Immobilization of heavy metals (Pb, Cu, Cr, Zn, Cd, Mn) in the mineral additions containing concrete composites. AB - The presented work determines the level of heavy metals (Pb+2, Cu+2, Zn+2, Cr+6, Cd+2, Mn+2) immobilization in the composites produced using Ordinary Portland Cement (OPC) as well as of binders containing large amount of mineral additives in its composition-siliceous fly ash (FA), fluidized bed combustion ash (FFA) and ground granulated blast furnace slag (GGBFS). Heavy metals were introduced to cementitious materials in the form of soluble salts as well as components of hazardous wastes (medical ash, metallurgical dust). It has been stated, that the level of heavy metals immobilization is combined with composites composition. Majority of analyzed heavy metals, added to binders' composition in the form of heavy metal salts achieves high level of immobilization, in mortar based on binder with 85% GGBFS and 15% OPC. The lowest immobilization level was reached for chromium Cr+6 added to hardening mortars as Na2Cr2O72H2O. The level ranges from 85.97% in mortars made on blended binder (20% OPC, 30% FFA and 50% GGBFS) to 93.33% in mortar produced on OPC. The increase of the so-called immobilization degree with time of hardened material maturing was found. This should be attributed to the pozzolanic or pozzolanic/hydraulic properties of components used; their effect on microstructure of hardened material is also important. Mineral additions enter the hydration reactions in the mixtures and favor the formation of specific microstructure promoting the immobilization of hazardous elements. PMID- 18423860 TI - Simultaneous removal of copper and lead ions from a binary solution by sono sorption process. AB - The aim of this work is to compare the simultaneous sorption of copper and lead ions from a binary aqueous solution in the presence and in the absence of ultrasound. The experiments under sonication were carried out by 20-kHz apparatus. Results indicated that the removal of copper and lead ions from a binary aqueous solution was greater in the presence of ultrasound than in control method. The removal of these ions was examined by varying experimental conditions such as the amount of sorbent, contact time, and temperature. In addition, the competitive sorption of ions was considered with different concentrations of each ion under the constant total concentration. The Langmuir isotherm model fits adequately the experimental data. In point of kinetics, the second-order kinetic model describes the sorption process for both ions. It was found that more than 90% of the lead and 60% of the copper ions was removed in less than 2min from the solution under sonication. The thermodynamic parameters such as DeltaG degrees , DeltaH degrees , and DeltaS degrees were determined for the sorption of Cu(2+) and Pb(2+) from the temperature dependence of the sorption process. PMID- 18423861 TI - Effect of calcination temperatures on microstructures and photocatalytic activity of tungsten trioxide hollow microspheres. AB - Tungsten trioxide hollow microspheres were prepared by immersing SrWO4 microspheres in a concentrated HNO3 solution, and then calcined at different temperatures. The prepared tungsten oxide samples were characterized by X-ray diffraction, X-ray photoelectron spectroscopy, Fourier transform infrared spectra, differential thermal analysis-thermogravimetry, UV-visible spectrophotometry, scanning electron microscopy, N2 adsorption/desorption measurements. The photocatalytic activity of the samples was evaluated by photocatalytic decolorization of rhodamine B aqueous solution under visible-light irradiation. It was found that with increasing calcination temperatures, the average crystallite size and average pore size increased, on the contrary, Brunauer-Emmett-Teller-specific surface areas decreased. However, pore volume and porosity increased firstly, and then decreased. Increasing calcination temperatures resulted in the changes of surface morphology of hollow microspheres. The un-calcined and 300 degrees C-calcined samples showed higher photocatalytic activity than other samples. At 400 degrees C, the photocatalytic activity decreased greatly due to the decrease of specific surface areas. At 500 degrees C, the photocatalytic activity of the samples increased again due to the junction effect of two phases. PMID- 18423862 TI - Renal expression of polyomavirus large T antigen is associated with nephritis in human systemic lupus erythematosus. AB - We have demonstrated that glomerular expression of polyomavirus large T antigen (T-ag) in a binary tetracycline-regulated T-ag transgenic mouse model (i) terminated tolerance for nucleosomes, (ii) released complexes of nucleosomes and T-ag to the microenvironment from dead cells, and (iii) that these complexes bound induced anti-nucleosome antibodies and finally (iv) that they associated with glomerular membranes as immune complexes. This process may be relevant for human lupus nephritis, since productive polyomavirus infection is associated with this organ manifestation. Here, we compare nephritis in the T-ag transgenic mouse with nephritis in human SLE. Glomerular sections were analysed by transmission electron microscopy, immune electron microscopy (IEM) and by co-localization IEM and TUNEL IEM assays to compare morphological changes, composition of immune complexes and formation of nucleosome-T-ag complexes. Affinity of nucleosome-T-ag complexes for glomerular collagen IV and laminin was determined by surface plasmon resonance (SPR). Analyses revealed electron dense structures in both human and murine kidney samples. These EDS were shown to contain T-ag, DNA and histones, indicating that extra-cellular chromatin may originate from polyomavirus infected cells in human kidneys. SPR analyses demonstrated high affinity of nucleosomes and nucleosome-T-ag complexes for collagen IV and laminin. Complexes of nucleosomes, T-ag and anti-T-ag and anti-dsDNA antibodies bind glomerular membranes and contribute to the evolution of lupus nephritis in human SLE. PMID- 18423863 TI - Neuropeptides, neurogenic inflammation and complex regional pain syndrome (CRPS). AB - This review explains symptoms and nature of neuropeptide signaling and its importance for clinical symptoms of CRPS. Neurogenic inflammation regularly accompanies excitation of primary afferent nociceptors. It has two major components-plasma extravasation and vasodilatation. The most important mediators are the calcitonin gene-related peptide (CGRP) and substance P (SP). After peripheral trauma immune reaction (e.g. cytokines) and the attempts of the tissue to regenerate (e.g. growth factors) sensitize nociceptors and amplify neurogenic inflammation. This cascade of events has been demonstrated in rat models of CRPS. Clinical findings in these animals strongly resemble clinical findings in CRPS, and can be prevented by anti-cytokine and anti-neuropeptide treatment. In CRPS patients, there is meanwhile also plenty of evidence that neurogenic inflammation contributes to clinical presentation. Increased cytokine production was demonstrated, as well as facilitated neurogenic inflammation. Very recently even "non-inflammatory" signs of CRPS (hyperhidrosis, cold skin) have been linked to neuropeptide signaling. Surprisingly, there was even moderately increased neurogenic inflammation in unaffected body regions. This favors the possibility that CRPS patients share genetic similarities. The future search for genetic commonalities will help us to further unravel the "mystery" CRPS. PMID- 18423864 TI - Evaluation of morphine analgesia and motor coordination in mice following cortex specific knockout of the N-methyl-D-aspartate NR1-subunit. AB - Studies have shown that N-methyl-D-aspartate (NMDA) receptors play a critical role in morphine analgesia and motoric processes at different levels of the central nervous system. In this study, we used cortex-specific NR1 knockout (KO) mice (C57BL/6 strain) to elucidate the role of cortical NMDA receptors in morphine analgesia and motor coordination. On post-natal day 20, mice (CTL and KO) received vehicle (saline) or morphine (10 mg/kg) and paw withdrawal latency (PWL) to a noxious thermal stimulus was measured. On post-natal day 21, motor coordination was measured using the rotating pole test. No differences in KO mice were found with respect to PWL following administration of saline or morphine (p>0.05). However, sex-dependent differences were found in motor coordination, with male KO mice showing a greater motor impairment in the rotating pole test than female KO mice (p<0.05). The present results demonstrate that NMDA receptors are involved in both the analgesic effects of morphine and motor coordination, with the existence of sex-related differences in motor coordination. PMID- 18423865 TI - Viral vector-mediated blockade of the endocrine stress-response modulates non spatial memory. AB - Stress results in the release of glucocorticoids (GCs) which at high levels, impair performance on hippocampus-dependent tasks. Estrogen is neurotrophic and can rescue stress-induced memory impairments. Here we report the use of a viral vector to overexpress a chimeric gene (ER/GR) that converts the deleterious effects of glucocorticoids into beneficial estrogenic effects. A short immobilization stress regimen was sufficient to impair non-spatial memory. In contrast, viral vector-mediated overexpression of ER/GR in the dentate gyrus of the hippocampus protected against stress-induced impairments of non-spatial memory. These data add to the growing evidence that increasing estrogenic signaling can protect against the impairing effects of stress on non-spatial memory. PMID- 18423866 TI - Interlimb coupling strength scales with movement amplitude. AB - The relation between movement amplitude and the strength of interlimb interactions was examined by comparing bimanual performance at different amplitude ratios (1:2, 1:1, and 2:1). For conditions with unequal amplitudes, the arm moving at the smaller amplitude was predicted to be more strongly affected by the contralateral arm than vice versa. This prediction was based on neurophysiological considerations and the HKB model of coupled oscillators. Participants performed rhythmic bimanual forearm movements at prescribed amplitude relations. After a brief mechanical perturbation of one arm, the relaxation process back to the initial coordination pattern was examined. This analysis focused on phase adaptations in the unperturbed arm, as these reflect the degree to which the movements of this arm were affected by the coupling influences stemming from the contralateral (perturbed) arm. The thus obtained index of coupling (IC) reflected the relative contribution of the unperturbed arm to the relaxation process. As predicted IC was larger when the perturbed arm moved at a larger amplitude than did the unperturbed arm, indicating that coupling strength scaled with movement amplitude. This result was discussed in relation to previous research regarding sources of asymmetry in coupling strength and the effects of amplitude disparity on interlimb coordination. PMID- 18423867 TI - Spinal injection of IL-2 or IL-15 alters mechanical and thermal withdrawal thresholds in rats. AB - IL-2 and IL-15 were tested for effects on responses to mechanical or thermal stimuli when spinally administered to male Sprague-Dawley rats with surgically implanted intrathecal catheters. Restricted doses of both IL-2 and IL-15 produced increased responsiveness to mechanical stimulation of the hindpaws. This effect lasted up to 48 h. IL-2 had biphasic effects on thermal responses whereas IL-15 produced thermal hypalgesia alone. These effects dissipated within 24h. These results suggest that IL-2 and IL-15 may participate in the generation of hyperalgesia in some pain conditions. PMID- 18423868 TI - Diabetes and the tortuosity of vessels of the bulbar conjunctiva. AB - OBJECTIVES: Diabetes is associated with loss of capillaries and macrovessel dilation in the conjunctiva, similar to well-known vessel changes in the retina. However, little is known about the effect of diabetes on the tortuosity of vessels of the conjunctiva. The authors examined the tortuosity of conjunctival vessels in participants with and without diabetes. DESIGN: Case-control study. PARTICIPANTS AND CONTROLS: Fifty-three patients with diabetes (17 with type 1 diabetes, 36 with type 2 diabetes) and 60 controls (all aged 20-94 years). METHODS: Digital red-free images of conjunctivae were analyzed using an automated computer algorithm to identify vessel axes and to quantify vessel tortuosity. Differences in vessel tortuosity were adjusted for age, gender, blood pressure, and smoking status. MAIN OUTCOME MEASURES: Tortuosity was expressed in units of curve energy (the square of the radian angular change between subsequent locations identified by the algorithm, standardized by vessel length). RESULTS: A longer duration of diabetes was associated with a reduction in overall vessel tortuosity (-2.8%; 95% confidence interval [CI], -4.3% to -1.3% per decade). This inverse association was driven by changes in larger vessels (40 microm in width or more), whereas increased tortuosity was observed in capillary sized vessels (<25 microm, 4.0%; 95% CI, -0.2% to 8.2% per decade). Compared with controls, those with type 1 diabetes (median duration of disease, 26 years) showed a 17.9% increase (95% CI, 4.7% to -31.0%) in capillary tortuosity. Conversely, those with type 1 diabetes showed a 7% decrease (95% CI, -11.8% to -2.3%) in tortuosity among vessels 40 to 80 microm or less in size and a 26.8% decrease (95% CI, 66.2% to 12.7%) in the fewer number of vessels more than 80 microm in size compared with controls. Similar, but smaller differences were seen in those with type 2 diabetes with shorter duration of diabetes (median, 7 years). CONCLUSIONS: Macrovessel dilation associated with diabetes may result in vessel engorgement and straightening, especially among those with longer durations of disease. Increased tortuosity associated with diabetes among conjunctival capillaries mirrors established vessel changes observed in the retina. Conjunctival angiopathy associated with diabetes may contribute to susceptibility to anterior eye disease among patients with diabetes. PMID- 18423869 TI - CFH and LOC387715/ARMS2 genotypes and treatment with antioxidants and zinc for age-related macular degeneration. AB - OBJECTIVE: To determine if CFH and LOC387715/ARMS2 genotypes influence treatment response to AREDS-type nutritional supplementation with antioxidants and zinc. DESIGN: Retrospective analysis of participants in a randomized, controlled clinical trial, the Age-Related Eye Disease Study (AREDS). PARTICIPANTS AND/OR CONTROLS: Eight hundred seventy-six AREDS study participants who were considered at high risk for developing advanced age-related macular degeneration (AMD). METHODS: Using DNA extracted from venous blood of 876 white participants in AREDS categories 3 and 4, that is, those considered to be at high risk for progression to advanced AMD, the authors genotyped for the single nucleotide polymorphisms in the CFH (Y402H, rs1061170) and LOC387715/ARMS2 (A69S, rs10490924) genes. The authors performed adjusted unconditional logistic regression analysis and assessed interactions of these genotypes to determine the relationship between CFH and LOC387715/ARMS2 genotype and treatment with antioxidants plus zinc. MAIN OUTCOME MEASURES: Interaction between genetic variants and treatment response as determined by progression from high-risk to advanced AMD. RESULTS: Progression occurred in 264 of 876 patients from AREDS category 3 (intermediate AMD) to category 4 or 5 (unilateral or bilateral advanced AMD, respectively), or from category 4 to category 5. A treatment interaction was observed between the CFH Y402H genotype and supplementation with antioxidants plus zinc (CC; P = 0.03). An interaction (P = 0.004) was observed in the AREDS treatment groups taking zinc when compared with the groups taking no zinc, but not in groups taking antioxidants compared with those taking no antioxidants (P = 0.59). There were no significant treatment interactions observed with LOC387715/ARMS2. CONCLUSIONS: The findings of this study indicate that an individual's response to AREDS supplements may be related to CFH genotype. This could have clinical relevance by predicting treatment outcome and potentially preventing unwanted side effects in those who may not benefit. Corroboration of these analyses is needed before considering modification of current management. This is among the first pharmacogenetic studies to suggest interaction between genotype and treatment. PMID- 18423870 TI - En bloc perfluorodissection for tractional retinal detachment in proliferative diabetic retinopathy. AB - OBJECTIVE: En bloc perfluorodissection (EBPD) is a surgical dissection technique that facilitates removal of epiretinal membranes and the posterior hyaloid. It is carried out by injecting perfluorocarbon liquid between the retina and the posterior hyaloid to separate the epiretinal tissues from the subjacent retina. The objective of this study was to describe the technique and to demonstrate the applicability of using EBPD during vitrectomy in eyes with tractional retinal detachment (TRD) in proliferative diabetic retinopathy (PDR). DESIGN: Prospective, interventional case series. PARTICIPANTS: Fifty-seven patients (eyes) with TRD in PDR participated in the study. METHODS: The author performed EBPD in 57 eyes (consecutive patients) during vitrectomy for TRD in severe PDR. Mean age of the patients was 42 years (range, 23-84 years). Mean surgical time was 50 minutes (range, 40-75 minutes). The group of patients was followed up for a mean of 18 months (range, 8-26 months). MAIN OUTCOME MEASURES: Best-corrected visual acuity (BCVA), retinal status, and complications at the end of follow up. RESULTS: None of the patients have developed ocular hypertension or undue inflammation, and anatomic success-accompanied by visual improvement (> or =2 Early Treatment Diabetic Retinopathy Study [ETDRS] lines) in 70.1% (40/57) occurred in 100% (57/57) of eyes. In 14 eyes (24.5%), BCVA remained stable, and in 3 eyes (5.2%), BCVA decreased (> or =2 ETDRS lines). Final BCVA was 20/50 or better in 25%, between 20/60 and 20/400 in 47%, and worse than 20/400 in 28%. Complications included phthisis bulbi in 1 (1.7%) eye, iatrogenic retinal breaks in 4 (7%) eyes, vitreous hemorrhage requiring another procedure in 4 (7%) eyes, and cataract in 15 (26.3%) eyes. CONCLUSIONS: This report demonstrates the applicability of using EBPD during vitrectomy in eyes with TRD in PDR. PMID- 18423872 TI - Selective modulation of nociceptive processing due to noise distraction. AB - This study investigates the effects of noise distraction on the different components and sources of laser-evoked potentials (LEPs) whilst attending to either the spatial component (localisation performance task) or the affective component (unpleasantness rating task) of pain. LEPs elicited by CO2 laser stimulation of the right forearm were recorded from 64 electrodes in 18 consenting healthy volunteers. Subjects reported either pain location or unpleasantness, in the presence and absence of distraction by continuous 85 dBa white noise. Distributed sources of the LEP peaks were identified using Low Resolution Electromagnetic Tomography (LORETA). Pain unpleasantness ratings and P2 (430 ms) peak amplitude were significantly reduced by distraction during the unpleasantness task, whereas the localisation ability and the corresponding N1/N2 (310 ms) peak amplitude remained unchanged. Noise distraction (at 310 ms) reduced activation in the anterior cingulate cortex (ACC) and precuneus during attention to localisation and unpleasantness, respectively. This suggests a complimentary role for these two areas in the control of attention to pain. In contrast, activation of the occipital pole and SII were enhanced by noise during the localisation and unpleasantness task, respectively, suggesting that the presence of noise was associated with increased spatial attentional load. This study has shown selective modulation of affective pain processing by noise distraction, indicated by a reduction in the unpleasantness ratings and P2 peak amplitude and associated activity within the medial pain system. These results show that processing of the affective component of pain can be differentially modulated by top-down processes, providing a potential mechanism for therapeutic intervention. PMID- 18423871 TI - Progression of myopia and high myopia in the early treatment for retinopathy of prematurity study: findings to 3 years of age. AB - PURPOSE: Examine the prevalence of myopia and high myopia, at 6 and 9 months postterm and 2 and 3 years postnatal in preterm children with birth weights < 1251 g who developed high-risk prethreshold retinopathy of prematurity (ROP) in the neonatal period and participated in the Early Treatment for ROP Study. DESIGN: Randomized controlled clinical trial. PARTICIPANTS: Four hundred one infants who developed prethreshold ROP and were determined to have a significant risk (>/=15%) of poor structural outcomes without treatment. Children underwent cycloplegic retinoscopy at examinations between 6 months postterm and 3 years' postnatal age. INTERVENTION: Eyes were randomized to receive treatment at high risk prethreshold ROP (early treated [ET]) or conventional management (CM), with treatment only if threshold ROP developed. MAIN OUTCOME MEASURES: Myopia (spherical equivalent >/= 0.25 diopters [D]) or high myopia (>/=5.00 D) at each visit. RESULTS: Prevalences of myopia were similar in treated eyes in the ET and CM groups, increasing from approximately 58% to 68% between 6 and 9 months, with little change thereafter. Both ET and CM eyes showed an increasing prevalence of high myopia, approximately 19% at 6 months and increasing 4% to 8% at successive examinations. Zone of ROP and presence or absence of plus disease had little effect on prevalence of myopia or high myopia between ages 6 months and 3 years. However, eyes with ROP residua (straightened temporal vessels or macular heterotopia) showed a higher prevalence of myopia and high myopia than eyes without residua. CONCLUSIONS: Approximately 70% of high-risk prethreshold ROP eyes were myopic in early childhood, and the proportion with high myopia increased steadily between ages 6 months and 3 years. Timing of treatment of high risk prethreshold ROP did not influence refractive error development. There was little difference in prevalence of myopia or high myopia between eyes with zone I and eyes with zone II ROP, nor between eyes with plus disease and eyes with no plus disease. However, prevalence of myopia and high myopia was higher in eyes with retinal residua of ROP than in eyes with normal-appearing posterior poles, highlighting the importance of follow-up eye examinations of infants who had prethreshold ROP. PMID- 18423873 TI - Acupuncture and the CNS: what can the brain at rest suggest? PMID- 18423874 TI - Neurotoxic and insecticidal properties of venom from the Australian theraphosid spider Selenotholus foelschei. AB - The present study is the first report on the bioactivity of venom from the Australian theraphosid spider Selenotholus foelschei. Venom from female specimens was used in all experiments. Adult spiders yielded an average of 2.2 mg dried venom per milking with a maximum yield of 7.2 mg. To evaluate the activity of pooled S. foelschei venom in invertebrates, a toxicity test in crickets (Acheta domesticus) was used. The results suggest the presence of several insecticidal toxins with different but synergistic modes of action, leading to a fast onset of paralysis as well as persistent paralysis and lethal effects (starting at 4 h after injection) in crickets. Vertebrate activity of S. foelschei venom was tested by using the isolated chick biventer cervicis nerve-muscle preparation. The venom produced a significant reduction in baseline tension (at a concentration of 10 microg/mL) and twitch height (at 0.2-10 microg/mL). Twitches were not restored after repeated washing. The response of the muscle to exogenous acetylcholine (1 mM) and carbachol (0.02 mM) was not reduced by the venom. These results indicate the presence of a vertebrate-active neurotoxin in S. foelschei that irreversibly blocks muscle twitches by acting either on voltage-activated Na(+)-channels or on other pre-synaptically located receptors. PMID- 18423875 TI - Optimal aperture sizes and positions for EMCD experiments. AB - The signal-to-noise ratio (SNR) in energy-loss magnetic chiral dichroism (EMCD)- the equivalent of X-ray magnetic circular dichroism (XMCD) in the electron microscope--is optimized with respect to the detector shape, size and position. We show that an important increase in SNR over previous experiments can be obtained when taking much larger detector sizes. We determine the ideal shape of the detector but also show that round apertures are a good compromise if placed in their optimal position. We develop the theory for a simple analytical description of the EMCD experiment and then apply it to dynamical multibeam Bloch wave calculations and to an experimental data set. In all cases it is shown that a significant and welcome improvement of the SNR is possible. PMID- 18423876 TI - Human metapneumovirus infection in an immunocompetent adult presenting as mononucleosis-like illness. AB - A 37-year-old man was presented with incidental findings of neutropenia, atypical lymphocytosis, thrombocytopenia and deranged liver parenchymal enzymes. Four days later, he developed fever, sore throat and cervical lymphadenopathy, compatible with mononucleosis-like illness (MLI). Polymerase chain reaction (PCR) and viral culture of the nasopharyngeal swab showed human metapneumovirus (hMPV). There was a >/=4-fold rise in IgG against hMPV. This is the first case report illustrating the natural clinical course of hMPV-related MLI. PMID- 18423877 TI - Anthelmintic activity of Lippia sidoides essential oil on sheep gastrointestinal nematodes. AB - Medicinal plants have been investigated for their anthelmintic properties and shown to be effective against eggs and larvae of gastrointestinal nematodes. The aim of this study was to evaluate the efficacy of the Lippia sidoides essential oil (LsEO) on sheep gastrointestinal nematodes. Initially, 44 naturally infected sheep were divided and treated with 200 microg kg(-1) ivermectin and 230 and 283 mg kg(-1) LsEO, respectively, plus the control. Fecal samples were collected from each animal to determine epg at 7, 14 and 21 days after treatment. In another test, 21 sheep were distributed and treated with 200 microg kg(-1) ivermectin, 283 mg kg(-1) LsEO and the control, respectively. Seven days after treatment, they were euthanized and necropsied to count and identify the nematodes from the abomasum, small and large intestines. In the first test, the efficacy of 230 and 283 mg kg(-1) LsEO and ivermectin was 38%, 45.9% and 40.2%, respectively, 7 days after treatment, and 30%, 54% and 39.6%, respectively, 14 days after treatment. In the second experiment, the respective efficacy of 283 mg kg(-1) LsEO and ivermectin was 56.9% and 34.4% against Haemonchus spp., and 39.3% and 63.6% against Trichostrongylus spp. PMID- 18423878 TI - Towards the standardization of the abbreviations of genus names of ticks (Acari: Parasitiformes: Ixodida). AB - An abbreviation system for genus names of ticks of the world is proposed. This abbreviation system adopts two letters (typically, the first two), as previously proposed for Culicidae and more recently for Phlebotominae. I hope this abbreviation system will be useful for anyone interested in tick taxonomy and related areas. PMID- 18423880 TI - Influence of mitochondrial enzyme deficiency on adult neurogenesis in mouse models of neurodegenerative diseases. AB - Mitochondrial defects including reduction of a key mitochondrial tricarboxylic acid cycle enzyme alpha-ketoglutarate-dehydrogenase complex (KGDHC) are characteristic of many neurodegenerative diseases. KGDHC consists of alpha ketoglutarate dehydrogenase, dihydrolipoyl succinyltransferase (E2k), and dihydrolipoamide dehydrogenase (Dld) subunits. We investigated whether Dld or E2k deficiency influences adult brain neurogenesis using immunohistochemistry for the immature neuron markers, doublecortin (Dcx) and polysialic acid-neural cell adhesion molecule, as well as a marker for proliferation, proliferating cell nuclear antigen (PCNA). Both Dld- and E2k-deficient mice showed reduced Dcx positive neuroblasts in the subgranular zone (SGZ) of the hippocampal dentate gyrus compared with wild-type mice. In the E2k knockout mice, increased immunoreactivity for the lipid peroxidation marker, malondialdehyde occurred in the SGZ. These alterations did not occur in the subventricular zone (SVZ). PCNA staining revealed decreased proliferation in the SGZ of E2k-deficient mice. In a transgenic mouse model of Alzheimer's disease, Dcx-positive cells in the SGZ were also reduced compared with wild type, but Dld deficiency did not exacerbate the reduction. In the malonate lesion model of Huntington's disease, Dld deficiency did not alter the lesion-induced increase and migration of Dcx-positive cells from the SVZ into the ipsilateral striatum. Thus, the KGDHC subunit deficiencies associated with elevated lipid peroxidation selectively reduced the number of neuroblasts and proliferating cells in the hippocampal neurogenic zone. However, these mitochondrial defects neither exacerbated certain pathological conditions, such as amyloid precursor protein (APP) mutation-induced reduction of SGZ neuroblasts, nor inhibited malonate-induced migration of SVZ neuroblasts. Our findings support the view that mitochondrial dysfunction can influence the number of neural progenitor cells in the hippocampus of adult mice. PMID- 18423881 TI - Involvement of stem cell factor and its receptor tyrosine kinase c-kit in pain regulation. AB - The c-kit receptor tyrosine kinase is expressed in a subpopulation of small- and medium-sized neurons of the dorsal root ganglia (DRG) and in the superficial layer of the spinal cord. Stem cell factor (SCF), a ligand of the c-kit receptor, induces neurite outgrowth from DRG and supports the survival of c-kit-expressing neurons. To clarify the possible function of the SCF/c-kit receptor system in the adult animal, we investigated the expression of c-kit receptor in the spinal cord and DRG in relation to pain by using H2C7, a newly developed anti-c-kit monoclonal antibody. S.c. and intrathecal injection of SCF markedly reduced the paw withdrawal threshold to mechanical stimuli and intrathecal SCF at 10 pg maximally induced mechanical allodynia in conscious mice. Intrathecal SCF also reduced the paw withdrawal latency to heat stimuli significantly but transiently. The c-kit receptor was co-expressed in 58.4% of calcitonin gene-related peptide (CGRP) -positive, but only 5.1% of isolectin B4-positive, DRG neurons. In the spinal cord, the c-kit receptor was detected in the superficial layer of the dorsal horn and co-localized there with CGRP in central terminals of DRG neurons. Selective elimination of unmyelinated C-fibers by neonatal capsaicin treatment resulted in marked reduction of the c-kit receptor and CGRP expression in the superficial layer of the spinal cord. Cell-size profiles showed that c-kit receptor expression was significantly up-regulated and down-regulated in medium sized DRG neurons after neonatal capsaicin treatment and nerve injury, respectively. These results suggest that the c-kit receptor is mainly expressed in peptidergic small-sized DRG neurons and may be involved in pain regulation both peripherally and centrally. PMID- 18423879 TI - Prevention and treatment of type 2 diabetes: current role of lifestyle, natural product, and pharmacological interventions. AB - Common complications of type 2 diabetes (T2D) are eye, kidney and nerve diseases, as well as an increased risk for the development of cardiovascular disease and cancer. The overwhelming influence of these conditions contributes to a decreased quality of life and life span, as well as significant economic consequences. Although obesity once served as a surrogate marker for the risk of T2D, we know now that excess adipose tissue secretes inflammatory cytokines that left unchecked, accelerate the progression to insulin resistance and T2D. In addition, excess alcohol consumption may also increase the risk of T2D. From a therapeutic standpoint, lifestyle interventions such as dietary modification and/or exercise training have been shown to improve glucose homeostasis but may not normalize the disease process unless weight loss is achieved and increased physical activity patterns are established. Furthermore, utilization of natural products may serve as a significant adjunct in the fight against insulin resistance but further research is needed to ascertain their validity. Since it is clear that pharmaceutical therapy plays a significant role in the treatment of insulin resistance, this review will also discuss some of the newly developed pharmaceutical therapies that may work in conjunction with lifestyle interventions, and lessen the burden of behavioral change as the only strategy against the development of T2D. PMID- 18423882 TI - Response patterns to sound associated with labeled globular/bushy cells in cat. AB - The mammalian cochlear nucleus (CN) consists of a diverse set of neurons both physiologically and morphologically that are involved in processing different aspects of the sound signal. One class of CN neurons that is located near the entrance of the auditory nerve (AN) to the CN has an oval soma with an eccentric nucleus and a short-bushy dendritic tree and is called a globular/bushy cell (GBC). They contact the principal cells of the medial nucleus of the trapezoid body (MNTB) with the very large calyx of Held that is one of the most secure synapses in the brain. Because MNTB cells provide an inhibitory input to the lateral superior olive (LSO), a structure purported to play a role in lateralizing high frequency sounds, GBC physiology is of great interest. Results were obtained with intracellular recording and subsequent labeling with neurobiotin of 32 GBCs along with a number of cells characterized extracellularly as likely GBCs in the cochlear nucleus (CN) of cat. Their poststimulus discharge response pattern to repeated tones varies from a primarylike pattern, i.e. similar to the AN, to a primarylike pattern with a 0.5-2 ms notch after the initial spike, to an onset pattern with a low-sustained rate. They can represent low frequency tones and amplitude modulated signals exceptionally well with a temporal code. PMID- 18423883 TI - Associations between the human growth hormone-releasing hormone- and neuropeptide Y-immunoreactive systems in the human diencephalon: a possible morphological substrate of the impact of stress on growth. AB - Previous studies revealed that stress is a pivotal factor in the regulation of growth. Psychological harassment may result in psychosocial dwarfism with delayed puberty, short stature and depression. Growth hormone (GH) secretion is suppressed by stress, possibly via the attenuation of growth hormone-releasing hormone (GHRH) secretion. However, the morphological substrate of this phenomenon has not been elucidated yet. Since neuropeptide Y (NPY) levels in the plasma is increased by administration of various stressors, the common consensus is that NPY plays a crucial role in the stress response. In the present study, we examined the putative juxtapositions between the NPY- and GHRH-immunoreactive (IR) systems in the human hypothalamus using double-label immunohistochemistry. Our findings revealed that the majority of the GHRH-IR perikarya formed intimate associations with NPY-IR fiber varicosities. The majority of these juxtapositions were found in the infundibular nucleus/median eminence where NPY-IR fiber varicosities often covered a significant surface area of the GHRH neurons. Since the juxtapositions between the GHRH-IR perikarya and NPY-IR fiber varicosities may be functional synapses, they may represent the morphological substrate of stress-suppressed GH secretion. The large number of contacting elements indicates that NPY plays a pivotal role in GH release, and may be considered as a major factor in the attenuation of growth by stress in humans. PMID- 18423884 TI - Click train encoding in primary and non-primary auditory cortex of anesthetized macaque monkeys. AB - We studied encoding of temporally modulated sounds in 28 multiunits in the primary auditory cortical field (AI) and in 35 multiunits in the secondary auditory cortical field (caudomedial auditory cortical field, CM) by presenting periodic click trains with click rates between 1 and 300 Hz lasting for 2-4 s. We found that all multiunits increased or decreased their firing rate during the steady state portion of the click train and that all except two multiunits synchronized their firing to individual clicks in the train. Rate increases and synchronized responses were most prevalent and strongest at low click rates, as expressed by best modulation frequency, limiting frequency, percentage of responsive multiunits, and average rate response and vector strength. Synchronized responses occurred up to 100 Hz; rate response occurred up to 300 Hz. Both auditory fields responded similarly to low click rates but differed at click rates above approximately 12 Hz at which more multiunits in AI than in CM exhibited synchronized responses and increased rate responses and more multiunits in CM exhibited decreased rate responses. These findings suggest that the auditory cortex of macaque monkeys encodes temporally modulated sounds similar to the auditory cortex of other mammals. Together with other observations presented in this and other reports, our findings also suggest that AI and CM have largely overlapping sensitivities for acoustic stimulus features but encode these features differently. PMID- 18423885 TI - Demonstration of decreased activity of dorsal raphe nucleus neurons in depressed suicidal patients by the AgNOR staining method. AB - BACKGROUND: Suicide and depression are closely related yet distinct phenomena. In both these phenomena, research has focused on central serotonergic system disturbances. The dorsal raphe nucleus (DRN) is the main source of serotonergic innervation of limbic structures crucial for the regulation of emotionally influenced behaviour. METHODS: The study was carried out on paraffin-embedded brains from 23 depressed patients (12 suicides and 11 non-suicides) and 26 matched controls without mental disorders. The karyometric parameters of DRN neurons were evaluated by the AgNOR silver staining method. RESULTS: The significant effect of suicide on the nuclear area was found in the cumulative analysis of all DRN subnuclei (ANOVA, P=0.032). A decreased mean value of this parameter was observed in the suicides group versus controls (t-test, P=0.032). This effect was especially pronounced in the violent suicide victims (t-test, P=0.001), who also demonstrated a decreased AgNOR area versus controls (t-test, P=0.007). No significant effect of depression or polarity on AgNOR parameters was found. LIMITATIONS: A major limitation of this study is relatively small case number. A further limitation is given by the lack of data on drug exposure across the whole life span. CONCLUSION: Our findings suggest that hypoactivity of DRN neurons is a distinct phenomenon in depression, specific only for suicidal subgroup of depressed patients. PMID- 18423886 TI - Polymorphism of the interleukin 1 receptor antagonist (IL1Ra) gene and placental abruption. AB - Candidate genes with a possible involvement in placental abruption are mainly those related to thrombophilia and preeclampsia. Some reports have shown by placental histologic investigation that increased risk of placental abruption is associated with prolonged inflammation. The polymorphic allele A2 in the gene coding for interleukin 1 receptor antagonist (IL1Ra) has been associated in various diseases of autoimmune or inflammatory nature. In obstetrics, previous research data has linked altered IL1Ra protein production with placental pathology and some severe pregnancy complications. In this study, we have determined whether IL1Ra gene polymorphism is associated also with an increased risk of placental abruption. The study involved 116 women with placental abruption and 112 healthy control pregnant women who were genotyped for polymorphism of the IL1Ra gene. The genotype and allele frequencies were assessed between the two groups and also compared with those in the general population. The frequency of the A2 allele was 28.0% among cases and 33.0% in controls (p=0.29), both similar to that in the general population (28.9%). In addition, the genotype distribution of IL1Ra polymorphisms was similar in both groups. Interestingly, there were a relatively higher number of cases with allele A3 (n=4; 1.7%) compared with the controls (0.4%) and the general population (1.0%) but the difference was not statistically significant. We conclude that there is no significant difference in IL1Ra polymorphisms between patients with and without placental abruption. PMID- 18423887 TI - The HLA-G 14bp gene polymorphism and decidual HLA-G 14bp gene expression in pre eclamptic and normal pregnancies. AB - Trophoblast expression of the non-classical MHC, HLA-G, is considered essential for feto-maternal immune tolerance and successful placentation in pregnancy. The HLA-G 14bp polymorphism in the 3'-untranslated region (UTR) of the HLA-G gene has been reported to be associated with development of pre-eclampsia (PE). In this study, maternal (peripheral blood, n=54) and fetal (cord blood, n=57) HLA-G 14bp genotypes have been determined by PCR in pre-eclamptic and normal pregnancies. In addition, HLA-G 14bp gene expression in decidua basalis (n=59) was analyzed by RT PCR. The pre-eclamptic syndrome was neither associated with the HLA-G 14bp genotype (maternal or fetal), nor with altered decidual HLA-G 14bp gene expression. Furthermore, the HLA-G 14bp mRNA expressed in decidua basalis was of fetal origin and all potential transcripts, as predicted from the fetal HLA-G 14bp genotype, were expressed. In contrast to previous findings, we found no correlation between the HLA-G 14bp polymorphism and fetal growth. In conclusion, the fetal HLA-G 14bp genotype is reflected in the decidual HLA-G mRNA splice form profile, but does not appear to be associated with increased risk for development of PE. PMID- 18423888 TI - S100B in underweight and weight-recovered patients with anorexia nervosa. AB - Anorexia nervosa (AN) commonly arises during adolescence, leading to interruptions of somatic and psychological development as well as to cortical atrophy and reductions of brain volume. While most brain changes shift towards normal with weight restoration, it is not certain whether they are related to the loss of brain cells, neuropil or merely due to fluid shifts. We measured S100B serum concentrations and psychometric characteristics in 34 patients with acute AN, 19 weight-recovered patients and 35 healthy control women (HCW). Plasma tryptophan and leptin levels were determined as markers for malnutrition and neuroendocrine adaptation to semi-starvation. Peripheral S100B concentrations of acute and former AN patients were not elevated and not statistically different from HCW. BMI, peripheral leptin levels and measures of psychopathology as well as executive cognitive functioning did not correlate with S100B. Plasma tryptophan was positively related to S100B. Our results are in line with our previous findings showing unaltered GFAP and NSE plasma levels in patients with acute AN. Together they do not support hypotheses comprising the degeneration of glial or neuronal cells to explain common signs of brain atrophy in patients with acute AN. PMID- 18423889 TI - Transcytosis of F4 fimbriae by villous and dome epithelia in F4-receptor positive pigs supports importance of receptor-dependent endocytosis in oral immunization strategies. AB - Very few antigens have been described that induce an intestinal immunity when given orally. Our laboratory demonstrated that oral administration of isolated F4 (K88) fimbriae of Escherichia coli to F4-receptor positive (F4R(+)) pigs induces protective mucosal immunity against challenge infection. However, presence of F4 receptors (F4R) on villous enterocytes is a prerequisite for inducing the immune response, as no F4-specific antibody-secreting cells (ASC) can be induced in F4R( ) pigs. In this study, the in vivo binding of isolated F4 fimbriae (F4) to the gut epithelium was examined in F4R(+) and F4R(-) pigs. It was further investigated whether binding of F4 to the F4R results in endocytosis in and translocation across the gut epithelium using microscopy. F4 did not adhere to the intestinal epithelium of F4R(-) pigs, whereas it strongly adhered to the villous epithelium and the follicle-associated epithelium (FAE) of the jejunum and ileum of F4R(+) pigs. Following binding to F4R, F4 was endocytosed by villous enterocytes, follicle-associated enterocytes and M cells. Transcytosis of F4 across the epithelium resulted in the appearance of F4 in the lamina propria and dome region of the jejunal and ileal PP. This is the first study showing transcytosis of fimbriae across the gut epithelium. This receptor-dependent transcytosis can explain the success of F4 fimbriae as oral immunogen for inducing protective immunity in F4R(+) pigs strengthening the importance of receptor-dependent endocytosis and translocation in oral vaccine strategies. Further identification of the receptor responsible for this transport is in progress. PMID- 18423890 TI - Quantitation of canine regulatory T cell populations, serum interleukin-10 and allergen-specific IgE concentrations in healthy control dogs and canine atopic dermatitis patients receiving allergen-specific immunotherapy. AB - Canine atopic dermatitis (AD) shares many clinical and immunological similarities with human AD. Regulatory T cells (Treg) are a distinct lineage of T lymphocytes with various immunosuppressive properties including the down-regulation of allergic inflammation associated with IgE production. Antigen-induced Treg typically regulate immune homeostasis via productions of cytokines such as interleukin-10. Given the immunological similarities with human AD, it is likely that Tregs and the cytokines they produce play an important role in diseases of dogs as well. A cross-reactive FoxP3 antibody was used to identify a subset of CD4(+) T cells in the blood of both healthy dogs and dogs with atopic dermatitis undergoing immunotherapy over a year period. There was no significant difference in the Treg percentage over time in the healthy dogs. The immunotherapy group showed a significant increase in Treg percentage at 6, 9, and 12 months when compared to the healthy dogs. For the immunotherapy group, the mean Treg percentage at the beginning of the study was 4.94+/-0.71 and 10.86+/-2.73 at the completion. A commercially available ELISA kit was also used to quantitate the concentration of IL-10 in the serum of the same subsets of dogs. There was no significant difference in the IL-10 concentrations over time in the healthy dogs. The immunotherapy group showed a significant increase in serum IL-10 concentrations at 6, 9, and 12 months when compared to the control group. The mean serum IL-10 concentration at the initiation of immunotherapy was 20.40+/ 3.52ngL(-1) and 37.26+/-15.26ngL(-1) at the completion of the study. The immunotherapy group also showed a significant decrease in serum IgE levels over the 1-year treatment period for specific allergens identified during ASIT. We conclude from these studies that similar to humans undergoing immunotherapy, increasing Treg populations likely play a significant role in the success of this particular type of therapy for atopic dermatitis and other allergic conditions. PMID- 18423891 TI - Nanodelivery of myelin: a therapeutic option for multiple sclerosis. PMID- 18423892 TI - Cochlear implantation and connexin expression in the child with keratitis ichthyosis-deafness syndrome. AB - We report one child with keratitis-ichthyosis-deafness (KID) syndrome. The child showed sparse, short scalp and body hairs, ichthyotic skin, bilateral sensorineural hearing loss, and visual loss. These are associated with a nucleotide substitution, 148G>A, which results in amino acid change in codon 50 of gap junctional protein connexin (Cx) 26. Immunohistochemistry of skins showed up-regulation of Cx26 and down-regulation of Cx43. He has benefited from cochlear implantation for hearing even though low visual function and skin problems with high risk of infections. This is the very rare report about Cx expression in skins and cochlear implantation in KID syndrome. PMID- 18423893 TI - Body position and obstructive sleep apnea in 8-12-month-old infants. AB - AIM: To determine the association between body position and obstructive events during sleep as determined by polysomnography (PSG) in infants of ages 8-12 months with obstructive sleep apnea (OSA). METHODS: Consecutive nocturnal polysomnograms (NPSGs) of 50 children ages 8-12 months old referred to the sleep disorders center between 1 January 2003 and 1 June 2006 for possible sleep disordered breathing were retrospectively reviewed. Data on total obstructive apnea index (AI), total obstructive apnea-hypopnea index (AHI), AI by body position, AHI by body position, rapid eye movement (REM) and non-REM sleep AHI and REM and non-REM AI were recorded. RESULTS: The mean age was 9.5+/-1.9S.D. months and 46% of the patients were females. There were no significant differences between the mean non-supine AHI (2.0+/-5.1) and supine AHI (2.5+/ 5.4), p=0.63. When comparing specific body positions, there were also no significant differences between the mean supine AHI (2.5+/-5.4), prone AHI (2.9+/ 7.3), left-lateral decubitus AHI (1.1+/-6.1), or the right-lateral decubitus AHI (2.5+/-7.6), p=0.71. Additionally, there were also no significant differences between the mean non-supine AI (0.7+/-2.9) and supine AI (1.4+/-3.0), p=0.23, and no differences between the supine AI (0.7+/-2.9), prone AI (1.0+/-2.9), left lateral decubitus AI (0.3+/-2.9) or the right-lateral decubitus AI (1.1+/-3.0), p=0.44. Children spent an average of 50% of their total sleep time supine. OSA was significantly worse in REM sleep as compared to non-REM sleep (REM AHI 4.3+/ 7.3 versus non-REM AHI 1.4+/-3.9, p=0.015; REM AI 5.1+/-4.9 versus non-REM AI 1.5+/-4.9, p<0.001). Mean time in REM sleep was 26% (range 5-42%). CONCLUSIONS: There was no significant effect of body position on sleep-disordered breathing in 8-12 months old infants, although REM sleep represented a significant risk factor for OSA. PMID- 18423894 TI - Is increased colon subepithelial collagen layer thickness in diabetic patients related to collagenous colitis? An immunohistochemical study. AB - In this study, we evaluated immunohistochemically whether increased thickness of the colon subepithelial collagen layer in diabetic patients relates to collagenous colitis. A total of 100 patients (25 in each group) were included in this study. There were diabetic patients with chronic diarrhea in the first group, diabetic patients without chronic diarrhea in the second group, non diabetic patients with chronic diarrhea in the third group, and control patients in the fourth group. The endoscopic biopsy specimens were obtained from the rectum, sigmoid colon, and descending colon. The thickness of the subepithelial collagen layer was measured using the ocular micrometer method. The immunohistochemical staining was performed with type 1 collagen and fibronectin antibody. The thickness of the colon subepithelial collagen layer in diabetic patients with or without diarrhea was significantly greater than that in control patients. This thickened subepithelial collagen layer in diabetic patients was stained with fibronectin antibody, but not with type 1 collagen antibody in the immunohistochemical study. These immunohistochemical staining characteristics were not similar to those in collagenous colitis, but were similar to those in normal subjects. Increased colon subepithelial collagen layer thickness in diabetic patients does not relate to collagenous colitis. PMID- 18423895 TI - Endolymphatic sac tumor (aggressive papillary tumor of middle ear and temporal bone): report of two cases with analysis of the VHL gene. AB - Endolymphatic sac tumor (Heffner tumor) (ELST) is a very rare nonmetastasizing, locally aggressive low-grade adenocarcinoma of endolymphatic sac origin, which is linked to von-Hippel-Lindau disease (VHLD). VHLD is an autosomal dominant disorder characterized by an inherited genetic abnormality of the VHL gene located on the short arm of chromosome 3 (3p26-p25). VHL gene mutations have been shown both in ELSTs associated with VHLD and in sporadic cases. Because of the rarity of ELST, only a small number of cases have been subjected to molecular genetic analysis. We have encountered two patients with ELST, one of whom presented with a medical and family history of VHLD. The second was a sporadic case, the patient having no symptoms of VHLD. The tissues obtained from Heffner tumor and cerebellar hemangioblastoma from the patient with inherited VHLD possess a point mutation in exon 1 of VHL gene. This mutation is a C to T exchange at position 194, resulting in amino acid exchange S65L. No mutation was found in any of the three exons analyzed and in the exon-intron junctions of the VHL gene in the sporadic case. PMID- 18423896 TI - Cyclin-dependent kinase 9: a key transcriptional regulator and potential drug target in oncology, virology and cardiology. AB - Unlike other CDKs, CDK9 does not regulate the cell cycle but promotes RNA synthesis in genetic programmes for cell growth, differentiation and viral pathogenesis. It is becoming clear that CDK9 inhibition contributes to the anticancer activity of most CDK inhibitors under clinic investigation. CDK9 was discovered in the context of HIV research because retroviruses hijack host transcription and CDK9 inhibitors might become specific antiretroviral agents, particularly as they might prevent drug resistance. Myocardial hypertrophy is a risk factor in congestive heart failure and is characterised by derepressed CDK9 activity. CDK9 inhibitors, thus, can find therapeutic application in cardiology. Although there are strong signs that CDK9 inhibition would be a useful therapeutic strategy in all three indications, the lack of selective inhibitors has so far confounded clinical development. Here we give an overview of the validity of CDK9 as a drug target and of the current knowledge of this kinase and its inhibitors. PMID- 18423897 TI - Flavin-containing monooxygenases: mutations, disease and drug response. AB - Flavin-containing monooxygenases (FMOs) metabolize numerous foreign chemicals, including drugs, pesticides and dietary components and, thus, mediate interactions between humans and their chemical environment. We describe the mechanism of action of FMOs and insights gained from the structure of yeast FMO. We then concentrate on the three FMOs (FMOs 1, 2 and 3) that are most important for metabolism of foreign chemicals in humans, focusing on the role of the FMOs and their genetic variants in disease and drug response. Loss-of-function mutations of FMO3 cause the disorder trimethylaminuria. More common variants that decrease enzyme activity are associated with increased drug efficacy. Most humans are homozygous for a nonsense mutation that inactivates FMO2. But a substantial proportion of sub-Saharan Africans express functional FMO2 and, thus, are predicted to respond differently to drugs and other foreign chemicals. PMID- 18423898 TI - Transitive inference in non-human animals: an empirical and theoretical analysis. AB - Transitive inference has long been considered one of the hallmarks of human deductive reasoning. Recent reports of transitive-like behaviors in non-human animals have prompted a flourishing empirical and theoretical search for the mechanism(s) that may mediate this ability in non-humans. In this paper, I begin by describing the transitive inference tasks customarily used with non-human animals and then review the empirical findings. Transitive inference has been demonstrated in a wide variety of species, and the signature effects that usually accompany transitive inference in humans (the serial position effect and the symbolic distance effect) have also been found in non-humans. I then critically analyze the most prominent models of this ability in non-human animals. Some models are cognitive, proposing for instance that animals use the rules of formal logic or form mental representations of the premises to solve the task, others are based on associative mechanisms such as value transfer and reinforcement and non-reinforcement. Overall, I argue that the reinforcement-based models are in a much better empirical and theoretical position. Hence, transitive inference in non-human animals should be considered a property of reinforcement history rather than of inferential processes. I finalize by shedding some light on some promising lines of research. PMID- 18423899 TI - Development of a high-throughput Alamar blue assay for the determination of influenza virus infectious dose, serum antivirus neutralization titer and virus ca/ts phenotype. AB - FluMist is an intranasal influenza live vaccine containing two Influenza A strains (currently H1N1 and H3N2) and one B strain (Yamagata or Victoria lineage). Characterization of the vaccine requires determination of the median tissue culture infectious dose (TCID(50)) titer, serum antivirus neutralization titer and vaccine cold adapted/temperature sensitive (ca/ts) phenotype. Visual cytopathic effect (CPE) readings are used widely in viral assays, but these are subjective and labor intensive. In response to the need for an efficient, inexpensive and high-throughput assay, a 96-well microplate assay was developed that uses Alamar blue dye staining as a replacement for CPE observation in the determination of influenza virus infectious dose, serum antivirus neutralization titer and virus ca/ts phenotype. Relative operating characteristic curves verified that there was a clear distinction between the fluorescence readings of the Alamar blue stained CPE positive and CPE negative wells. Virus titer was determined by use of both Alamar blue staining and CPE-based TCID(50) assays for wild-type and FluMist influenza vaccine strains as well as a plasmid-rescued influenza FluMist A strain containing a H5N1 derived hemmaglutinin and neuramidinase. Correlation of the two assays was measured by regression analysis and resulted in R(2) values of 0.814 (Influenza A), 0.983 (Influenza B) and 1.000 (H5N1), respectively. Serum microneutralization as well as virus ca/ts phenotype assays also showed a high concordance between readings based on CPE observation and Alamar blue staining. The Alamar blue dye assay is user friendly, environmentally safe and sensitive. Also, it is adaptable to automation, which could provide a high-throughput platform for analysis of pre-clinical and clinical samples. PMID- 18423900 TI - The developmental antecedents of illicit drug use: evidence from a 25-year longitudinal study. AB - BACKGROUND: The present study examined the developmental antecedents of illicit drug use and abuse/dependence. METHODS: A 25-year prospective longitudinal study of the health, development, and adjustment of a birth cohort of 1265 New Zealand children. Measures included assessments of adolescent and young adult illicit drug use and abuse/dependence; cannabis use to age 25; measures of parental adjustment; measures of exposure to childhood sexual abuse, physical abuse, and interparental violence; novelty-seeking; childhood and early adolescent adjustment and substance use; and affiliation with substance-using peers. RESULTS: Illicit drug use and abuse/dependence from ages 16 to 25 were significantly associated (all p values<.05) with a range of parental adjustment measures; exposure to abuse in childhood; individual factors; and measures of childhood and early adolescent adjustment. Analyses using repeated measures logistic regression models suggested that parental illicit drug use, gender, novelty-seeking, and childhood conduct disorder predicted later illicit drug use and abuse/dependence. Further analyses revealed that these pathways to illicit drug use and abuse/dependence were mediated via cannabis use, affiliation with substance-using peers, and alcohol use during ages 16-25. CONCLUSIONS: The current study suggested that the illicit drug use and abuse/dependence were associated with a range of early life circumstances and processes that put individuals at greater risk of illicit drug use and abuse/dependence. However, the use of cannabis in late adolescence and early adulthood emerged as the strongest risk factor for later involvement in other illicit drugs. PMID- 18423902 TI - Inhibitory effects of some derivatives of glycyrrhizic acid against Epstein-Barr virus infection: structure-activity relationships. AB - Glycyrrhizic acid (18beta-GL or GL) is a herbal drug with a broad spectrum of antiviral activities and pharmacological effects and multiple sites of action. Previously we showed that GL inhibits Epstein-Barr virus (EBV) infection in vitro by interfering with an early step of the EBV replication cycle (possibly attachment/penetration). Here we tested the effects of 15 GL derivatives against EBV infection by scoring the numbers of cell expressing viral antigens and quantifying EBV DNA copy numbers in superinfected Raji cells. The derivatives were made either by transformation of GL on carboxyl and hydroxyl groups or by conjugation of amino acid residues into the carbohydrate part. We identified seven compounds active against EBV and all showed dose-dependent inhibition as determined by both assays. Among these active compounds, the introduction of amino acid residues into the GL carbohydrate part enhanced the antiviral activity in three of the seven active compounds. However, when Glu(OH)-OMe was substituted by Glu(OMe)-OMe, its antiviral activity was completely abolished. Introduction of potassium or ammonium salt to GL reduced the antiviral activity with no significant effect on cytotoxicity. The alpha-isomer (18alpha-GL) of 18beta-GL was as potent as the beta-form, but its sodium salt lost antiviral activity. The metabolic product of GL, 18beta-glycyrrhetinic acid (18beta-GA or GA), was 7.5 fold more active against EBV than its parental compound GL but, concomitantly, exhibited increased cytotoxicity resulting in a decreased therapeutic index. PMID- 18423901 TI - Optimising the benefits of unobserved dose administration for stable opioid maintenance patients: follow-up of a randomised trial. AB - BACKGROUND: The registration of combination buprenorphine/naloxone, a formulation designed to reduce risk of diversion, has led some Australian jurisdictional authorities to allow treatment without direct observation of dosing for stable, opioid-dependent patients. AIM: To compare two approaches (1) initiating treatment with observed dosing, then allowing patients who demonstrate stability to change to unobserved dosing; or (2) initiating patients with unobserved dosing, subsequently requiring those who fail to stabilize to change to observed treatment. METHODS: This study builds on an RCT comparing efficacy of observed and unobserved treatment at 3 months. At the conclusion of the RCT, clinically "stable" subjects were allocated to continue without observed dosing, while those who did not demonstrate stability were allocated to observed dosing. Subjects were followed for a further 3 months. Primary end-point was retention in treatment. RESULTS: Of 119 subjects randomised, 70 were retained in treatment to 3 months. Forty-five stable subjects were allocated to unobserved dosing, 25 to observation. Unstable subjects allocated to observed treatment were more likely to drop out thereafter (OR 2.14, 95% CI 1.09-4.19). There was a non-significant trend for people initiated with observed dosing to be better retained during the allocation phase; at 6 months, 13 subjects (22%) from the original unobserved group, and 22 (34%) from the observed group, were retained in treatment (chi2=2.10, 1 df, p=0.15). CONCLUSIONS: Withdrawal of unobserved doses led to marked attrition from treatment. If access to unobserved dosing is to be restricted to stable patients, it appears preferable to initiate dosing with observation and allow unobserved doses for people who successfully stabilize, than to initiate with unobserved doses and transfer unstable patients to observation. PMID- 18423903 TI - Characterization and localization of ORFF gene from the LD1 locus of Leishmania donovani. AB - The Leishmania genome project has identified new genes at a rapid rate. The 32.8 megabase haploid genome of Leishmania major (Friedlin strain) is published and the comparative analysis of genome sequences of two other species, Leishmania infantum and Leishmanai braziliensis has been done. The haploid genome of Leishmania major (Friedlin strain) has around 8272 protein-coding genes, of which only 36% can be ascribed a putative function. Out of these open reading frames around 910 Leishmania major genes have no orthologs in the other two Tritryp genomes. These "Leishmania -restricted" genes hold a potential as novel drug targets and potential vaccine candidates. Open reading frame, ORFF, is a single copy gene located on the chromosome 35 as a part of the multigene LD1 locus. Indirect immunofluorescence study and creation of ORFF-GFP fusion showed that ORFF is localized in the DNA containing compartments of Leishmania donovani, the nucleus and the kinetoplast. In order to characterize ORFF gene of L. donovani, we have created ORFF over-expressors and single allele deletion mutants by homologous replacement strategy. ORFF is likely to be an important gene for the parasite growth since results from over-expression studies and characterization of ORFF heterozygous knockout mutants reveal marked alterations in the cell cycle phenotype compared to the wild-type parasites. Flowcytometry based cell cycle analysis showed selective increase in the DNA synthetic phase of the ORFF over expressors and a subversion of the same in heterozygous knockouts of ORFF suggesting its potential role in cell cycle progression. PMID- 18423904 TI - Imprinting of idiosyncratic experience in cortical sensory maps: neural substrates of representational remodeling and correlative perceptual changes. AB - Over the past 30 years, extensive research has been conducted in the field of cortical plasticity, under the impetus of seminal studies showing that the mature brain retains a capacity to reorganize the morphological and functional architecture of its neural circuits in order to adapt to environmental changes and mediate functional recovery following injury. Much effort has been focused on determining how idiosyncratic experience translates into molecular, structural and physiological changes in the sensory and motor representations embedded within cortical networks. The wealth of data generated by a broad spectrum of experimental manipulations has allowed unprecedented progress in our understanding of the physiological processes and neuroplasticity mechanisms underlying cortical representational remodeling. The objective of the present review is to put various facets of cortical map plasticity into perspective so as to examine possible links between changes occurring at multiple scales of the neural organization of the mature brain. The main focus is on neural substrates that mediate the instructive influence of experience and behavioral context on cortical reorganization, and perceptual correlates of representational remodeling. PMID- 18423906 TI - On the other hand: dummy hands and peripersonal space. AB - Where are my hands? The brain can answer this question using sensory information arising from vision, proprioception, or touch. Other sources of information about the position of our hands can be derived from multisensory interactions (or potential interactions) with our close environment, such as when we grasp or avoid objects. The pioneering study of multisensory representations of peripersonal space was published in Behavioural Brain Research almost 30 years ago [Rizzolatti G, Scandolara C, Matelli M, Gentilucci M. Afferent properties of periarcuate neurons in macaque monkeys. II. Visual responses. Behav Brain Res 1981;2:147-63]. More recently, neurophysiological, neuroimaging, neuropsychological, and behavioural studies have contributed a wealth of evidence concerning hand-centred representations of objects in peripersonal space. This evidence is examined here in detail. In particular, we focus on the use of artificial dummy hands as powerful instruments to manipulate the brain's representation of hand position, peripersonal space, and of hand ownership. We also review recent studies of the 'rubber hand illusion' and related phenomena, such as the visual capture of touch, and the recalibration of hand position sense, and discuss their findings in the light of research on peripersonal space. Finally, we propose a simple model that situates the 'rubber hand illusion' in the neurophysiological framework of multisensory hand-centred representations of space. PMID- 18423905 TI - Chronic administration of DHA and UMP improves the impaired memory of environmentally impoverished rats. AB - Living in an enriched environment (EC) during development enhances memory function in adulthood; living in an impoverished environment (IC) impairs memory function. Compounds previously demonstrated to improve memory among IC rats include CDP-choline and uridine monophosphate (UMP). Brain phosphatidylcholine (PC) synthesis utilizes both the uridine formed from the metabolism of exogenous CDP-choline and UMP, and the choline formed from that of CDP-choline. It also uses the polyunsaturated fatty acid (PUFA) DHA, a precursor for the diacylglycerol incorporated into PC. DHA administration also improves cognition in young and aged rodents and humans; its effects on cognitively impaired IC rats have not been characterized. We have thus examined the consequences of administering DHA (300 mg/kg) by gavage, UMP (0.5% in the diet), or both compounds on hippocampal- and striatal-dependent forms of memory among rats exposed to EC or IC conditions for 1 month starting at weaning, and consuming a choline-containing diet. We observe that giving IC rats either dietary UMP or gavaged DHA improves performance on the hidden version of the Morris water maze (all P<0.05), a hippocampal-dependent task; co-administration of both phosphatide precursors further enhances the IC rats' performance on this task (P<0.001). Neither UMP nor DHA, nor giving both compounds, affects the performance of EC rats on the hidden version of the Morris water maze (P>0.05), nor the performance by IC or EC rats on the visible version of the Morris water maze (all P>0.05), a striatal-dependent task. We confirm that co-administration of UMP and DHA to rats increases brain levels of the phosphatides PC, PE, SM, PS, PI, and total brain phospholipid levels (all P<0.05), and show that rearing animals in an enriched environment also elevates brain PC, PS, and PI levels (all P<0.01) and total brain phospholipids (P<0.01) compared with their levels in animals reared in an IC environment. These findings suggest that giving DHA plus UMP can ameliorate memory deficits associated with rearing under impoverished conditions, and that this effect may be mediated in part through enhanced synthesis of brain membrane phosphatides. PMID- 18423907 TI - Phylogenetic background, virulence gene profiles, and genomic diversity in commensal Escherichia coli isolated from ten mammal species living in one zoo. AB - Three hundred commensal Escherichia coli recovered from healthy herbivorous, carnivorous, and omnivorous mammals from one zoo were characterized for their phylogenetic origin, intestinal virulence gene (VG) prevalence, and genomic diversity. The phylogenetic structure of the E. coli (groups A, B1, B2, and D) from the herbivores was homogenous, with a prevailing representation of group B1. In the carnivores and omnivores, the phylogenetic diversity was species specific with a higher representation of group A compared to the herbivores. Of 16 intestinal VGs in the whole set, 8 were detected and they formed 13 VG profiles. In the herbivores, all the VG-positive isolates belonged to group B1 and harboured the genes eaeA, eastI, ehxA, stx1, and stx2, which separately or in combination formed 8 VG profiles. In the carnivores and omnivores, the VG positive isolates frequently belonged to group A and harboured the estI and estII genes or a combination of eastI and estI, forming three VG profiles. Single genes cnf2, in group B2, and eastI, in group D, were found. Similarity analysis of pulsed-field gel electrophoresis (PFGE) patterns revealed closer relatedness between the isolates from carnivores and omnivores than those from herbivores. The comparison between the prevalence of phylogenetic groups and the phylogenetic origin of VG-positive isolates in the examined E. coli suggested, that E. coli from group B1 in herbivores and E. coli from group A rather than B1 in carnivores and omnivores are "best adapted" to the host organism. The groups revealed different preferences in the acquisition and maintenance of intestinal VGs. PMID- 18423908 TI - Characterization of an erythrocytic virus in the family Iridoviridae from a peninsula ribbon snake (Thamnophis sauritus sackenii). AB - A wild peninsula ribbon snake (Thamnophis sauritus sackenii) in Florida was found to have hypochromic erythrocytes containing two different types of inclusions: purple granular inclusions, and pale orange or pink crystalloid inclusions that were round, oval, rectangular, or hexagonal in shape. Transmission electron microscopy revealed hexagonal or pleomorphic, homogenous inclusions and enveloped particles morphologically consistent with a member of the Iridoviridae. Histopathology of the animal revealed necrotizing hepatitis consistent with sepsis. Consensus PCR was used to amplify a 628-bp region of iridoviral DNA dependent DNA polymerase. Bayesian and maximum likelihood phylogenetic analysis found that this virus was distinct from other known iridoviral genera and species, and may represent a novel genus and species. PMID- 18423909 TI - The home is the hub of health in very old age: Findings from the ENABLE-AGE Project. AB - The aim was to explore health in relation to the home as experienced by very old, single-living Swedish people. Applying a grounded theory approach, 40 men and women aged 80-89 were interviewed in their own homes. Data analysis revealed the main theme, "The home is the hub of health", comprising two categories, "The home as support for health", and "Having an inner driving force to maintain health". Health was described as being active and participating, and during the aging process the home became an increasingly important source of support. The older people were aware of their vulnerability, and knew that their life situation could change rapidly. Thus, health was always interpreted in relation to this. They had a strong inner driving force to maintain health, and within the home they challenged their capacity, and used different adaptive strategies targeting person-environment-activity transactions. Our findings imply the need for focusing on the opportunities for activity and participation in all interventions. They also challenge current national and international housing policy emphasizing the benefits of staying put, thus contributing to a more diverse view of what kind of housing arrangements are optimal for very old people. PMID- 18423911 TI - Consecutively developed late stent malappositions following the implantation of two different kinds of drug-eluting stents associated with spontaneous healing. AB - We describe here a 63-year-old woman, who consecutively developed a stent-related coronary aneurysm of left circumflex artery, which was associated with spontaneous healing, and sequential late stent malapposition of left anterior descending artery following the implantation of two different kinds of drug eluting stents, respectively. She did not experience any cardiac events for thirty eight months. PMID- 18423910 TI - Mathematical description of the uptake of hydrocarbons in jet fuel into the stratum corneum of human volunteers. AB - The objective of this research was to develop a mathematical description of uptake of aromatic and aliphatic hydrocarbons into the stratum corneum of human skin in vivo. A simple description based on Fick's laws of diffusion was used to predict the spatiotemporal variation of naphthalene, 1- and 2-methylnaphthalene, undecane, and dodecane in the stratum corneum of human volunteers. The estimated values of the diffusion coefficients for each chemical were comparable to values predicted using in vitro skin systems and biomonitoring studies. These results demonstrate the value of measuring dermal exposure using the tape-strip technique and the importance of quantifying of dermal uptake. PMID- 18423912 TI - Cria alpaca body weight and perinatal survival in relation to age of the dam. AB - The present study with alpacas determined effect of dam's age on body weight and survival of cria during the first week of life. Pregnant dams (n=424) and their crias were used in the study. Cria body weight (kg) was determined at time of placenta expulsion. Placenta weight and larger width were measured immediately after expulsion. Crias were monitored for the first week and a necropsy was performed if death occurred. Data were analyzed using analysis of variance. The body weight of crias at birth, the weight, and the largest width of placenta increased with age of the dam (P<0.05), reaching a peak at 9 years and then declined progressively. Placental efficiency also increased with the dam's age, and showed a bimodal shape, peaking at 6- and 11-year-old dams (P<0.05). Altogether, 398 crias survived and 26 died; of those 6 died of starvation, 5 of hypothermia, 4 were stillborn and the rest from other miscellaneous causes. More crias died from 2-year-old dams than from dams of any other age (P<0.05). In addition, dead crias had lesser body weights (6.4kg) than those of crias that survived (7.8kg, P<0.05). The weight and width of the placenta was similar for live and dead crias. PMID- 18423913 TI - Flow cytometric quantification of bacteria in vaginal swab samples self-collected by adolescents attending a gynecology clinic. AB - Bacterial vaginosis (BV) is an important risk factor in reproductive health outcomes, such as pre-term birth and sexually transmitted infections including HIV. However, its etiology, diagnosis and treatment remain poorly defined. We evaluated flow cytometry as a tool to quantify total bacterial cells in vaginal specimens self-collected longitudinally by adolescents. BV was diagnosed by Gram stain (criteria of Hay and Ison). Average flow cytometric counts of bacterial cell-units (BCU) was log(10) 8.04 per gram sample and was found to correlate with sample weight (p<0.0001). BV was frequently observed in this group, with 22 of 32 participants (69%) diagnosed with BV for at least one timepoint. Surprisingly, increased BCU was associated with normal Hay-Ison score (p=0.0003), even when adjusting for sample weight (p=0.02). Since presence and quantity of Lactobacillus defines normal vaginal microbiology (ie. absence of BV), this result indicates a possible bias towards dominance of Lactobacillus cells in measurements of "total" BCU. Increased BCU per gram was associated in multivariate analysis with longer self-reported time since last menstruation (p=0.004) and last sexual intercourse (p=0.007). Sperm was detected in 3 samples provided by those reporting sexual intercourse in the previous 24 h. Light scattering profiles of bacteria and vaginal cells in samples collected over time from an individual were often identical and distinct from other individuals. To our knowledge, this is the first description of flow cytometry for analysis of commensal bacteria in vaginal specimens. Further development may help to illuminate the complex dynamics of vaginal microbial communities underlying BV. PMID- 18423914 TI - Radiotherapy for 41 patients with stages I and II MALT lymphoma: a retrospective study. AB - PURPOSE: Mucosa-associated lymphoid tissue (MALT) lymphoma is a distinct disease with specific clinical and pathologic features that may affect diverse organs. We analyzed our recent experience with Stage I/II MALT lymphoma presenting in the stomach and other organs to assess the outcome following radiation therapy (RT) alone. PATIENTS AND METHODS: Forty-one patients with Stages I (37) and II (4) disease were treated between 2000 and 2006. Patients with transformed MALT were excluded. The median age was 60 years (range, 25-86 years), male: female ratio 1:1. Presenting sites included stomach, 11; orbital adnexa, 21; thyroid, 1; other head and neck, 3; small bowel, 3; skin, 1; and rectum, 1. Thirty-five patients (85%) received RT-alone and 6 (15%) received antibiotics followed by RT. RT dose was 30Gy in 20 fractions (fr) in all 41 patients. Mean follow-up time was 32.0 months (range, 2.1-162 months). RESULTS: A first complete response was achieved in all 41 patients. Only one patient died from bile duct carcinoma at 22 months from the start of irradiation for conjunctiva MALT lymphoma without recurrence of lymphoma. The other 40 patients were alive. Thirty-eight patients out of them were alive without recurrence. One patient with a duodenal lymphoma had a recurrence in non-irradiated distant sites at 1 month. Another patient with a bilateral eye lid lymphoma had a recurrence within radiation field at 41 months. The absolute local control rate with radiation was 98% (40/41 patients). CONCLUSION: Localized MALT lymphomas have excellent prognosis following moderate dose RT (30Gy/20fr). PMID- 18423915 TI - MDM2 SNP 309 and p53 codon 72 polymorphisms are associated with the outcome of oral carcinoma patients receiving postoperative irradiation. AB - BACKGROUND AND PURPOSE: The p53 tumor suppression pathway is important in effects associated with radiotherapy. The mouse double minute 2 (MDM2) plays a pivotal role in this pathway by down regulating p53. A functional T-to-G polymorphism at nucleotide 309 in MDM2 promoter intron 1 (SNP309) has been identified which influenced transcription activity. A G-to-C SNP at p53 codon 72 results in an Arg/Pro polymorphism, which is associated with apoptosis induction potential and p53 mutation status. MATERIALS AND METHODS: We sequenced both MDM2 SNP309 and p53 codon 72 SNP in patients with oral squamous cell carcinoma (OSCC, n=189), oral submucosal fibrosis (OSF, n=70), and 116 controls. RESULTS: Neither MDM2 SNP309 nor p53 codon 72 SNP was associated with susceptibility to or the age at onset of OSCC or OSF. p53 codon 72 SNP Arg/Arg polymorphism was associated with the progression of OSCC, and the overall (OS) and disease-free survival (DFS) of irradiated patients. The MDM2 SNP309 G/G polymorphism was associated with poor OS in advanced OSCC, and the OS and DSF of irradiated patients. The combination of MDM2 SNP309 G/G and p53 codon 72 Arg/Arg polymorphism is associated with the worst OS and DFS. CONCLUSIONS: Advanced OSCC has high mortality and recurrence. We identified that both MDM2 SNP309 and p53 codon 72 SNP could be useful factors for evaluating the outcome of advanced OSCC treated with adjuvant radiation. PMID- 18423916 TI - A phase I trial of pre-operative radiotherapy for prostate cancer: clinical and translational studies. AB - BACKGROUND AND PURPOSE: Selected patients undergoing radical prostatectomy for localized prostate cancer can be at high-risk for pT3 disease and require subsequent radiotherapy. In a phase I trial, we investigated the feasibility of pre-operative radiotherapy for this patient subset. MATERIALS AND METHODS: Eligibility criteria were: T1/T2N0M0 tumors plus (i) Gleason >or=7, PSA>10 ng/ml and <35 ng/ml, or (ii), PSA >15 ng/ml and less <35 ng/ml (any Gleason). Patients received 25 Gy in five fractions of radiotherapy followed by radical prostatectomy. Trial endpoints included intra-operative morbidity and late toxicity following combined treatment. We also stained pre- and post-radiotherapy prostate samples for DNA damage response proteins. RESULTS: Between 2001 and 2004, 15 patients were entered on trial. Thirteen patients completed combined modality treatment. Only one patient had signs of intra-operative inflammation. No patient had post-operative complication. There was no severe late gastrointestinal toxicity. Late genitourinary toxicity consisted of severe urinary incontinence in 2 of 13 patients. From a translational standpoint, irradiated prostate tumor tissues had long-term activation of the CDK-inhibitor p21(WAF) associated with reduced cell proliferation. CONCLUSION: Intra-operative morbidity is low following short-course, pre-operative radiotherapy. A phase II trial is planned to fully document biochemical response with this combined modality approach. PMID- 18423917 TI - Swelling and diffusion studies of calcium polysaccharide gels intended for film coating. AB - The aim of this paper was to study the swelling and diffusion behaviors of calcium polysaccharide gel (CaPG) films prepared by an interfacial complexation technique, a new gel formation method that allowed calcium ions to diffuse from a source to form gel films with polysaccharide (i.e., alginate or pectin). The dynamic swelling behavior of CaPG films showed that swelling was a function of time. Most CaPG films showed a maximum amount of water absorption during the first few hours. The films swelled less in water and acidic media but extensively swelled in 0.1M NaCl. The rehydration of the dry films in the acidic media or the 0.1M NaCl solution also lead to the extraction of most of the calcium ions from the CaPG within 4h or less. Partitioning and diffusion of a model drug, theophylline (TPL), were measured through CaPG films equilibrated in different media. The partition and diffusion coefficients of TPL through CaPG films were found to vary, depending upon polysaccharide type, concentration and equilibration medium. The results suggest that both partition and pore mechanisms operated concurrently in the transport of TPL through CaPG films equilibrated in different media. PMID- 18423918 TI - Affect, attention, or anticipatory arousal? Human blink startle modulation in forward and backward affective conditioning. AB - Affect modulates the blink startle reflex in the picture-viewing paradigm, however, the process responsible for reflex modulation during conditional stimuli (CSs) that have acquired valence through affective conditioning remains unclear. In Experiment 1, neutral shapes (CSs) and valenced or neutral pictures (USs) were paired in a forward (CS-->US) manner. Pleasantness ratings supported affective learning of positive and negative valence. Post-acquisition, blink reflexes were larger during the pleasant and unpleasant CSs than during the neutral CS. Rather than affect, attention or anticipatory arousal were suggested as sources of startle modulation. Experiment 2 confirmed that affective learning in the picture picture paradigm was not affected by whether the CS preceded the US. Pleasantness ratings and affective priming revealed similar extents of affective learning following forward, backward or simultaneous pairings of CSs and USs. Experiment 3 utilized a backward conditioning procedure (US-->CS) to minimize effects of US anticipation. Again, blink reflexes were larger during CSs paired with valenced USs regardless of US valence implicating attention rather than anticipatory arousal or affect as the process modulating startle in this paradigm. PMID- 18423919 TI - Uricosuric effect of Roselle (Hibiscus sabdariffa) in normal and renal-stone former subjects. AB - AIM OF THE STUDY: The Roselle (Hibiscus sabdariffa) was investigated for its uricosuric effect. MATERIALS AND METHODS: A human model with nine subjects with no history of renal stones (non-renal stone, NS) and nine with a history of renal stones (RS) was used in this study. A cup of tea made from 1.5 g of dry Roselle calyces was provided to subjects twice daily (morning and evening) for 15 days. A clotted blood and two consecutive 24-h urine samples were collected from each subject three times: (1) at baseline (control); (2) on days 14 and 15 during the tea drinking period; and (3) 15 days after the tea drinking was stopped (washout). Serum and 24-h urinary samples were analyzed for uric acid and other chemical compositions related to urinary stone risk factors. RESULTS: All analyzed serum parameters were within normal ranges and similar; between the two groups of subjects and among the three periods. Vis-a-vis the urinary parameters, most of the baseline values for both groups were similar. After taking the tea, the trend was an increase in oxalate and citrate in both groups and uric acid excretion and clearance in the NS group. In the RS group, both uric acid excretion and clearance were significantly increased (p<0.01). When the fractional excretion of uric acid (FEUa) was calculated, the values were clearly increased in both the NS and SF groups after the intake of tea and returned to baseline values in the washout period. These changes were more clearly observed when the data for each subject was presented individually. CONCLUSIONS: Our data demonstrate a uricosuric effect of Roselle calyces. Since the various chemical constituents in Roselle calyces have been identified, the one(s) exerting this uricosuric effect need to be identified. PMID- 18423920 TI - Different commercial yeast strains affecting the volatile and sensory profile of cava base wine. AB - 36 semi-industrial fermentations were carried out with 6 different yeast strains in order to assess differences in the wines' chemical and volatile profile. Two of the tested strains (Y3 and Y6) showed the fastest fermentation rates throughout 3 harvests and on 2 grape varieties. The wines fermented by three of the tested strains (Y5, Y3 and Y4) stand out for their high amounts of esters and possessed the highest fruity character. Wines from strains producing low amounts of esters and high concentrations of medium chain fatty acids, higher alcohols and six-carbon alcohols were the least appreciated at the sensory analysis. The data obtained in the present study show how the yeast strain quantitatively affects the final chemical and volatile composition of cava base wines and have repercussions on their sensory profile, independently of must variety and harvest year. PMID- 18423921 TI - Purification and characterisation of proteases A and D from Debaryomyces hansenii. AB - The proteases A (PrA; EC. 3.4.23.25) and D (PrD; EC. 3.4.24.37) of Debaryomyces hansenii CECT 12487 were characterised after their isolation by fractionation with protamine sulfate followed by three chromatographic separations, which included two anion exchange and one gel filtration chromatographic steps. The whole procedures for PrA and PrD resulted in 1349 and 2560 purification-fold with a recovery yield of 1.4 and 1.3%, respectively. PrA was active at acidic-neutral pH with an optimum pH between 5.0 and 6.0. PrD was active at neutral-basic pH with an optimum pH between 7.0 and 8.0. The molecular mass of the native PrA was 55 kDa and (being) 42 kDa in denaturing conditions. Polyclonal-antibodies raised against PrA from Saccharomyces cerevisiae cross-reacted with the corresponding PrA from D. hansenii. PrD showed a native molecular mass of 68 kDa and 65 kDa in denaturing conditions. PrA was an aspartic protease effectively inhibited by pesptatin A while PrD was classified as a metallo protease inhibited by 1,10 phenantroline and affected by some divalent cations such as zinc, cadmium and magnesium. The homology of the PrA to the lisosomal cathepsin D suggests its possible participation in the ripening of fermented meat products. PMID- 18423922 TI - A test of individuality of human external ear pattern: its application in the field of personal identification. AB - A preliminary study has been undertaken to test the individuality of human ear patterns and its probable use in profile view facial image recognition. Anthropometric measurements on 12 inter-landmark linear distances have been carried out for both left and right ears of 700 male and female individuals. A 12 dimensional feature space has been framed to represent each ear pattern as a feature vector with the measured inter-landmark distances as its components. The Euclidean distances in such a feature space amongst all possible pairs of ear patterns of both male and female individuals have been computed. Very few pairs have distances which fell below the safe distinction limit. The undistinguished pairs were further examined by direct superimposition of their images. None of the ear patterns is found to be exactly the same in morphology to the other. Even the left and right ears of the same individual are not identical. Thus, the present study reconfirms the individuality of ear patterns. For establishment of identity, a questioned profile view facial image can be compared with a control one on the basis of the ear pattern visible in it. PMID- 18423923 TI - [Isokinetic, functional and proprioceptive assessment of soccer players two years after surgical reconstruction of the anterior cruciate ligament of the knee]. AB - Injuries to the anterior cruciate ligament (ACL) of the knee are common in sport and are treated routinely in the world of sports medicine. In order to resume competitive sport in safety, it is important to know the actual level of performance achieved by the operated leg some time after the ligament repair. OBJECTIVES: The objective of this work was to evaluate the recovery of the operated leg in Tunisian sportsmen two years after surgery by using isokinetic testing (60 degrees s(-1)), the one-leg hop distance test (i.e. a functional activity that one often finds in sport) and proprioceptive assessment. A secondary objective was to detect any possible correlations between the various outcomes. MATERIALS AND METHOD: The ACL group was composed of 26 top-level Tunisian footballers having undergone ACL ligament repair. Two years after surgery, the following assessments were performed: an isokinetic test with an angular velocity of 60 degrees s(-1), a proprioceptive assessment in both active and passive modes and the functional one-leg hop distance test. RESULTS: Our investigations confirmed a muscle deficit of 16% in the hamstring muscles of the injured leg. In both active and passive modes, normal proprioception is observed at a flexion of 60 degrees and in total extension but not in a mid-way position. The performance level of the operated leg (in terms of distance) had an average symmetry value of 93.40+/-2.7%. CONCLUSION: Playing football appears to influence the isokinetic strength profile in sportsmen after ACL repair. The repair procedure leads to bilateral proprioceptive disruption at 15 degrees of knee flexion, on average. PMID- 18423924 TI - Sepsis associated encephalopathy in an infant with biliary atresia. AB - We describe a girl with sepsis-associated encephalopathy complicating biliary atresia. At 4 months of age, decreased consciousness and repetitive seizures of the left upper and lower extremities occurred in association with fever. Pseudomonas aeruginosa was cultured from blood, while bacterial culture was negative and cell counts were normal in cerebrospinal fluid. The interleukin-6 level in the cerebrospinal fluid was markedly elevated. MRI revealed unilateral subcortical white matter lesions in the right hemisphere. She was diagnosed as having sepsis-associated encephalopathy and was treated with dexamethasone and midazolam. She achieved normal psychomotor development until the last follow-up at 19 months of age, whereas mild atrophic changes were observed in the right hemisphere. PMID- 18423925 TI - The linear neuron as marker selector and clinical predictor in cancer gene analysis. AB - OBJECTIVE: The problem of gene selection has been extensively studied in a number of scientific works using various kinds of methods. However, the application of a linear neuron is a novel approach possessing several advantages. In this work we propose to study the behavior of such a linear neuron, appropriately adapted and trained to the problem of gene selection in the DNA microarray experiment. METHODS AND MATERIALS: We explore the proposed approach in terms of an accuracy evaluation criterion, which is used to assess the performance of the proposed methodology, but we also evaluate the produced results in terms of cluster quality and survival prediction. Cluster quality reflects the ability of the method to select differentially expressed genes, which in turn leads to better clustering and survival prediction. RESULTS: We directly compare the proposed methodology with RFE-SVM, a well known and broadly accepted method demonstrating remarkable performance on various data sets of clinical interest. CONCLUSIONS: Conducted computational experiments show that the proposed approach can be efficiently used within the field of gene selection producing high-quality results in terms of accuracy and robustness. PMID- 18423926 TI - A novel blood-cell-two-compartment model for transferring a whole blood time activity curve to plasma in rodents. AB - The term input function usually refers to the tracer plasma time activity curve (pTAC), which is necessary for quantitative positron emission tomography (PET) studies. The purpose of this study was to acquire the pTAC by independent component analysis (ICA) estimation from the whole blood time activity curve (wTAC) using a novel method, namely the FDG blood-cell-two-compartment model (BCM). This approach was compared to a number of published models, including linear haematocrit (HCT) correction, non-linear HCT correction and two exponential correction. The results of this study show that the normalized root mean square error (NRMSE) and the error of the area under curve (EAUC) for the BCM estimate of the pTAC were the smallest. Compartmental and graphic analyses were used to estimate the metabolic rate of the FDG (MR(FDG)). The percentage error for the MR(FDG) (PE(MRFDG)) was estimated from the BCM corrected pTAC and this was also the smallest. It is concluded that the BCM is a better choice when transferring wTAC into pTAC for quantification. PMID- 18423927 TI - Fast computation of approximate entropy. AB - The approximate entropy (ApEn) is a measure of systems complexity. The implementation of the method is computationally expensive and requires execution time analogous to the square of the size of the input signal. We propose here a fast algorithm which speeds up the computation of approximate entropy by detecting early some vectors that are not similar and by excluding them from the similarity test. Experimental analysis with various biomedical signals revealed a significant improvement in execution times. PMID- 18423928 TI - [Celiac axis lymphadenopathy and prognosis in esophageal cancer]. PMID- 18423929 TI - [A changing spectrum: mucosal damage and the clinical picture of adult celiac disease]. PMID- 18423931 TI - 14-3-3 regulates life span by both DAF-16-dependent and -independent mechanisms in Caenorhabditis elegans. AB - Caenorhabditis elegans life span, stress resistance and metabolism are regulated by the Insulin/IGF-1/DAF-2/DAF-16 pathway. DAF-16, a member of FOXO/Forkhead transcription factor family, can be targeted by 14-3-3 proteins to promote stress resistance. We have identified a 14-3-3 C. elegans homolog which promotes life span by both DAF-2-dependent and -independent mechanisms and by an unexpected DAF 16-independent mechanism. Our results demonstrate that C. elegans 14-3-3 proteins modulate stress-responsive genes throughout adulthood. In conclusion, 14-3-3 can be considered as an acute stress-responsive regulator as well as a sustained modulator of the Insulin/IGF-1/DAF-2/DAF-16 regulatory pathway in promoting life expectancy of growing old worms. PMID- 18423932 TI - Temperature, evapotranspiration and primary photochemical responses of apple leaves to hail. AB - The objective of this work was to examine immediate physiological plant responses to hail and subsequent recovery in terms of evapotranspiration, leaf temperature and primary photochemical processes using apple as a model crop. Thermal emission pictures were taken in darkness to avoid interference from stomatal movements; temperature gradients were identified in concentric rings around sites of hail injury, with a distinct drop in temperature of up to 2.3 degrees C in the center immediately after the induced hail injury. This was due to enhanced evapotranspiration from the injured tissue. Six to twelve minutes after hail injury, the initial decrease in leaf temperature partially reversed. Chlorophyll fluorescence kinetics of light-adapted leaves showed a dramatic decrease in effective photosynthetic electron transport rate (ETR), from 20.5 to 9.0 micromol electron m(-2)s(-1) within 5 min from hail injury, and a rapid recovery to 14.1 micromol electron m(-2)s(-1) within the next 5 min. After 7h, ETR partially recovered to 17.4 micromol electron m(-2)s(-1). An initial drop in non photochemical efficiency (NPQ) from 1.07 to 0.90 units within 5 min after hail injury was followed by a sharp increase to 1.67 units after another 5 min. During the next hour, NPQ gradually decreased to the initial level. This indicates increased thermal dissipation in photosystem II (PS II) as a protective mechanism against incident excessive energy in the leaves with closed stomata for 1h after hail injury. In contrast to the fluorescence kinetics of light-adapted leaves, maximum quantum yield Fv/Fm of PSII in the dark-adapted state remained unchanged at 0.79-0.81 relative units for the first 5 min after hail injury. Thereafter, Fv/Fm slowly declined to 0.75 within 1h, and to a trough of 0.73 at 3h. Seven hours after hail injury, Fv/Fm values were at 0.76, indicating partial recovery of PS II efficiency. The discrepancy in the dynamics of ETR and Fv/Fm responses may be explained by the formation of alternative electron sinks such as reactive oxygen species, particularly superoxides, which withdraw electrons from the photosynthetic transport, resulting in apparently higher values of calculated ETR. PMID- 18423930 TI - Combinatorial and rational approaches to polymer synthesis for medicine. AB - High-throughput, combinatorial methods have revolutionized small molecule synthesis and drug discovery. By combining automation, miniaturization, and parallel synthesis techniques, large collections of new compounds have been synthesized and screened. It is becoming increasingly clear that these same approaches can also assist the discovery and development of novel biomaterials for medicine. This review examines combinatorial and rational polymer synthesis for medical applications, including stem cell engineering and nucleic acid drug delivery. PMID- 18423933 TI - Physiological dissection of blue and red light regulation of apical dominance and branching in M9 apple rootstock growing in vitro. AB - This paper presents the results of the interaction of red light (R) with blue light (B), applied to shoots of M9 apple (Malus pumila paradisiaca Schmid) rootstock, on the regulation of stem growth, apical dominance and branching. These results are compared with the active form of phytochromes (PHYs) under monochromatic and dichromatic light treatments. The inhibition of internode elongation was determined by PHY photoequilibrium, with the additional effect of B, by means of a separate photoreceptor. The development of phytomers appeared to be primarily due to the active form of PHY, with a marginal effect from B. Shoot growth, which combines internode elongation and development of the phytomer, was highest under R and lowest under B and far red light (FR), showing the largely positive role of PHY photoequilibrium. Under FR, reduced stem elongation was due to the very small number of phytomers formed. Apical dominance was inhibited, while branching was increased under R, corresponding to the highest values of PHY photoequilibrium determined. Apical dominance was increased and branching was reduced by B, indicating a complex interaction between PHY and B receptor. In the shoot cluster system, photomorphogenic behavior was dependent on the time of exposure to the different light qualities. The information gained from the study will be helpful in improving the set up of in vitro growth light conditions, and in providing useful insights into research of the development of the woody plant canopy, an important factor in ecological plant communities. PMID- 18423934 TI - A pronounced light-induced zeaxanthin formation accompanied by an unusually slight increase in non-photochemical quenching: a study with barley leaves treated with methyl viologen at moderate light. AB - Light-induced deepoxidation of violaxanthin to antheraxanthin and zeaxanthin in plants is associated with the induction of pronounced xanthophyll-dependent non photochemical quenching (NPQ). To date, a misbalance between a high amount of zeaxanthin in thylakoid membranes and low NPQ has been explained by an absence of lumen acidification (e.g. when NPQ is measured in the dark after high light stress). In this study, we report that this misbalance can also be observed under moderate light. We found this result (deepoxidation state, DEPS, above 55% and NPQ approximately 0.9) in barley leaves treated with 10 microM methyl viologen (MV) under white light (100 micromol photonsm(-2)s(-1), photosynthetically active radiation (PAR), growth irradiance). The addition of MV at this moderate light did not accelerate electron transport in thylakoid membranes, and induced only slight oxidative stress (no lipid peroxidation, almost unchanged maximum yield of photosystem II photochemistry, a decrease in activity of ascorbate peroxidase, and an increase in that of glutathion reductase). We suggest that, in leaves treated under the conditions used here, the lumen acidification induced by light limited electron transport in thylakoid membranes was high enough to activate violaxanthin deepoxidase, but not sufficiently high to form the expected number of zeaxanthin-dependent quenching centers in photosystem II antennae. PMID- 18423935 TI - Normal anatomy, variants and pitfalls on shoulder MRI. AB - MR imaging of the shoulder is widely used for assessment of impingement and instability-related clinical conditions. The following review article demonstrates the normal anatomy, variations and classical pitfalls. In addition to classical pitfalls (sublabral hole, sublabral recess, Buford complex) the authors focus on a number of normal, bony, cartilaginous, ligamentous and tendinous structures that can simulate disease at the shoulder. In addition, ways to distinguish these pitfalls from true shoulder abnormalities are shown. PMID- 18423936 TI - (1)H MR spectroscopy in pediatrics. AB - Spectral appearance and concentrations of the most prominent metabolites are affected by brain development. This knowledge is essential for the detection of pathological changes in pediatric patients. This paper discusses specific conditions of MR spectroscopic examination of children and the effects of age on MR spectra quality and quantitation of the studied metabolites. Clinical examples show several diseases that are reflected in changes in (1)H MR spectra due to pathological alterations in the biochemical pathways of the observed metabolites. Attention is given to the main metabolites such as N-acetylaspartate, creatine/phosphocreatine, cholines, lactate, inositol, etc. PMID- 18423937 TI - PPAR-gamma expression in animals subjected to volume overload and chronic Urotensin II administration. AB - Activation of PPAR-gamma through the administration of glitazones has shown promise in preserving function following cardiac injury, although recent evidence has suggested their use may be contraindicated in the case of severe heart failure. This study tested the hypothesis that PPAR-gamma expression increases in a time dependent manner in response to chronic volume overload (VO) induced heart failure. Additionally, we attempted to determine what effect 4 week administration of Urotensin II (UTII) may have on PPAR-gamma expression. VO induced heart failure was produced in Sprague-Dawley rats (n=32) by aorta-caval fistula. Animals were sacrificed at 1, 4, and 14 weeks following shunt creation. In a separate set of experiments, animals were administered 300 pmol/kg/h of UTII for 4 weeks, subjected to 4 weeks of volume overload, or given UTII+VO. Densitometric analysis of left ventricular (LV) protein demonstrated PPAR-gamma expression was significantly ((*)p<0.05) upregulated at 4 and 14 weeks (31.5% and 37%, respectively) post-fistula formation compared to control values. PPAR-gamma activation was decreased in the 4 and 14 week (39.16% and 42.4%, respectively), but not in the 1-week animals, and these changes did not correlate with NF-kappaB activity. Animals given UTII either with or without VO demonstrated increased expression of PPAR-gamma as did animals subjected to 4 week VO alone. Animals given UTII either with or without VO had decreased activity vs. control. These data suggest PPAR-gamma may play a role in the progression of heart failure, however, the exact nature has yet to be determined. PMID- 18423938 TI - UVB photolysis of hydrocortisone 21-acetate. AB - Hydrocortisone 21-acetate (HCA) in methanol solution undergoes photodegradation under UVB light, as monitored by HPLC. Five main photoproducts have been isolated and characterized by means of NMR and mass spectroscopy. One of them derives from a Norrish I photoreaction which cleaves the C17-C20 bond of the steroid yielding the andro-derivative, a second product comes from a Yang-type photorearrangement which links C18 to C20 yielding a cyclobutane adduct. The former photoproduct, in turn, undergoes further photolysis giving rise to various photoproducts, of which three have been characterized. The first is a stereoisomer of the andro derivative, the others arise from the opening of the five-membered ring. HCA also proved photounstable in the solid state and in a commercial formulation for topical use, thus confirming the requirements of the Pharmacopeias for light protection of this drug. Indeed, experiments on LPS-stimulated THP-1 cells demonstrated the loss of anti-inflammatory activity when HCA was UVB photodegraded. The radical mechanism involved in HCA photolysis seems also responsible for the in vitro photohemolytic effect and lipid peroxidation induced by HCA in combination with UVB light. PMID- 18423939 TI - Analysis of the complexation of gemfibrozil with gamma- and hydroxypropyl-gamma cyclodextrins. AB - The interactions of gemfibrozil with gamma- and HP-gamma-cyclodextrin (CD) have been studied in aqueous solution by fluorescence and NMR spectroscopy and by solubility measurements and in the solid state by X-ray diffraction, thermal analysis and FTIR spectroscopy. The influence of the technique employed in the analysis of complexation is discussed. The fluorescence of gemfibrozil increased in the presence of gamma- and hydroxypropyl-gamma-cyclodextrin (HP-gamma-CD), especially with the later, because the inclusion of the aromatic ring in the cavity, evidenced by 1H NMR, has a protective effect on the excited state of the drug. The fluorescence enhancement allowed the determination of the binding constants at pH 2.8. Complexation was a both entropy and enthalpy driven process. The solubility diagrams obtained with gamma-CD and HP-gamma-CD were B(s) and A(L) type, respectively. The apparent stability constants calculated from the solubility data at 25 degrees C were compared with those obtained from the fluorescence assays. It was found that drug solubilization with gamma-CD involves other contributions together with the inclusion phenomena. Solid complexes of gemfibrozil with gamma-CD (and not with HP-gamma-CD) have been obtained by kneading, coevaporation and coprecipitation methods. The solid complexes crystallised in the channel structure, in a process involving the carboxyl and aryl-ether groups. PMID- 18423940 TI - A GSTM3 polymorphism associated with an etiopathogenetic mechanism in Alzheimer disease. AB - Brain-specific glutathione S-transferase Mu 3 (GSTM3) colocalizes with amyloid beta plaques in Alzheimer's disease (AD). A functional polymorphism rs7483 in GSTM3 may contribute to the decrease in GSTM3 expression in AD. The association of the rs7483 SNP with late-onset AD and mild cognitive impairment (MCI) was evaluated and the impact of a SNP background on gene expression was analyzed in blood mononuclear cells (BMC). The allelic association of the GSTM3 allele with AD was significant in women and in APOEvarepsilon4-negative stratum. A significant association was also found in both MCI and AD subjects with AD family history. GSTM3 transcript levels in BMC were lower in AD than in normal elderly controls, and the presence of the risk allele was associated with further mRNA reduction. Diminished GSTM3 mRNA levels correlated with decreased minichromosome maintenance deficient 3 (MCM3) mRNA levels in a diagnostic and SNP-dependent fashion. Reduced antioxidant defense and genome maintenance associated with the GSTM3 polymorphism suggest a common hub of regulatory networks which, when impaired, may lead to AD. PMID- 18423941 TI - Pain control for sternal fracture using an ultrasound-guided hematoma block. AB - In this case report, an ultrasound-guided hematoma block was performed in the Emergency Department (ED) for immediate and effective pain control in a patient suffering from a sternal fracture. This technique of anesthesia may allow safer and more effective analgesia and a more rapid discharge from the hospital or ED in selected cases. PMID- 18423942 TI - Larvicidal activities of some organotin compounds on mosquito larvae: a QSAR study. AB - Mosquitoes are not only the cause of nuisance by their bites but also transmit deadly diseases like malaria, filariasis, yellow fever, dengue, and Japanese encephalitis. In this paper, nine QSAR models were developed using different series of organotins with respect to their larvicidal activities against Aedes aegypti and Anopheles stephensi mosquito larvae. Internal [cross-validation (LOO q(2)), quality factor (Q), Fischer statistics (F), and Y-randomization] and external validation tests have validated all these QSAR models. QSAR results suggest that the two most important determinants for the toxicity are the hydrophobic (pi) and Hammett electronic (sigma(+)) parameters of the substituents, and the kill mechanism is different for these two species of mosquito larvae. On the basis of QSAR (6), nine compounds 4a-4i are suggested as potential synthetic targets. PMID- 18423943 TI - Molecular modeling and inhibition of phospholipase A2 by polyhydroxy phenolic compounds. AB - Phospholipases A(2) are enzymes responsible for the hydrolysis of membrane phospholipids that release arachidonic acid, which serves as substrate for pro inflammatory mediators, such as prostaglandins and leucotriens. The design of specific inhibitors for PLA(2) might help in the development of new anti inflammatory drugs. Polyhydroxy phenolic compounds, such as flavonoids, vitamin E, rosmarinic acid and aristolochic acid, are able to inhibit PLA(2) from different sources. Herein, we have studied the kinetic behavior and the capacity of inhibiting edema formation induced by PLA(2) of five different polyhydroxy phenolic compounds (two phenolic derivatives and three acetophenone hydroxylated derivatives) extracted from the venom of Crotalus adamanteus. The results showed that compounds 1,3-dihydroxy benzene, 1,3,5-trihydroxy benzene and 2,4,6 trihydroxy acetophenone were the most efficient in the inhibition of the enzymatic activity and edema induction by PLA(2). It was also verified that the number of hydroxyls in each molecule is not a limiting factor for the inhibition capacity of these compounds. Molecular modeling studies indicated that the most active compounds are linked to the amino acid Asp 49 and that they destabilize the coordination of the calcium atom, which is essential to the catalytic activity. The study of potential surfaces showed that there are conditions in which the potential values must be adequate for enzyme complex formation with polyhydroxy phenolic compounds. When the potential over the hydroxyl surfaces is very high, formation of stable complexes does not occur and the enzyme does not act intensely. These results might be helpful in the design of a drug that specifically inhibits PLA(2). PMID- 18423944 TI - A QSAR study for the cytotoxic activities of taxoids against macrophage (MPhi) like cells. AB - A series of taxoids (modified at the C-3'N position) have been used for the development of two QSAR models for their cytotoxic activities against macrophage (MPhi)-like cells J774.1 and J7.DEF3. QSAR results suggest that hydrophobic and steric features influence the inhibitory activity in a linear model. The ClogP values were not great enough to establish the upper limit of the inhibition. Internal [cross-validation, quality factor (Q), Fischer statistics (F), Y randomization, and lack of overfitting tests] and external validation tests have validated both the QSAR models. PMID- 18423946 TI - Construction and characterization of a second-generation pseudoinfectious West Nile virus vaccine propagated using a new cultivation system. AB - Safer vaccines are needed to prevent flavivirus diseases. To help develop these products we have produced a pseudoinfectious West Nile virus (WNV) lacking a functional C gene which we have named RepliVAX WN. Here we demonstrate that RepliVAX WN can be safely propagated at high titer in BHK cells and vaccine certified Vero cells engineered to stably express the C protein needed to trans complement RepliVAX WN growth. Using these BHK cells we selected a better growing mutant RepliVAX WN population and used this to generate a second-generation RepliVAX WN (RepliVAX WN.2). RepliVAX WN.2 grown in these C-expressing cell lines safely elicit strong protective immunity against WNV disease in mice and hamsters. Taken together, these results indicate the clinical utility of RepliVAX WN.2 as a vaccine candidate against West Nile encephalitis. PMID- 18423945 TI - Regardless of genotype, offspring of VIP-deficient female mice exhibit developmental delays and deficits in social behavior. AB - Pharmacological studies indicate that vasoactive intestinal peptide (VIP) may be necessary for normal embryonic development in the mouse. For example, VIP antagonist treatment before embryonic day 11 resulted in developmental delays, growth restriction, modified adult brain chemistry and reduced social behavior. Here, developmental milestones, growth, and social behaviors of neonates of VIP deficient mothers (VIP +/-) mated to VIP +/- males were compared with the offspring of wild type mothers (VIP +/+) mated to VIP +/+ and +/- males, to assess the contributions of both maternal and offspring VIP genotype. Regardless of their own genotype, all offsprings of VIP-deficient mothers exhibited developmental delays. No delays were seen in the offspring of wild type mothers, regardless of their own genotype. Body weights were significantly reduced in offspring of VIP-deficient mothers, with VIP null (-/-) the most affected. Regardless of genotype, all offspring of VIP-deficient mothers expressed reduced maternal affiliation compared with wild type offspring of wild type mothers; +/- offspring of wild type mothers did not differ in maternal affiliation from their wild type littermates. Play behavior was significantly reduced in all offsprings of VIP-deficient mothers. Maternal behavior did not differ between wild type and VIP-deficient mothers, and cross-fostering of litters did not change offspring development, indicating that offspring deficits were induced prenatally. This study illustrated that the VIP status of a pregnant mouse had a greater influence on the growth, development and behavior of her offspring than the VIP genotype of the offspring themselves. Deficiencies were apparent in +/+, +/- and -/- offspring born to VIP-deficient mothers; no deficiencies were apparent in +/- offspring born to normal mothers. These results underscore the significant contribution of the uterine environment to normal development and indicate a potential usefulness of the VIP knockout mouse in furthering the understanding of neurodevelopmental disorders with social behavior deficits such as autism. PMID- 18423947 TI - Beta-glycoglycosphingolipid-induced augmentation of the anti-HBV immune response is associated with altered CD8 and NKT lymphocyte distribution: a novel adjuvant for HBV vaccination. AB - BACKGROUND: Non-responsiveness towards the currently used hepatitis B virus (HBV) vaccine is a major problem in attempts to protect against HBV infection. Several methods have been tested to overcome the lack of an effective immune response towards HBV antigens. Adjuvants that augment the immunologic reaction are essential components of the vaccines. Beta-glycosphingolipids exert a natural killer T cell (NKT)-mediated immunomodulatory effect in various disorders. AIMS: The aim of the present study was to test the ability of these compounds to augment the immune response towards HBV antigens, making them potential adjuvants for HBV vaccines. Six groups of mice were injected with different formulations of an HBV vaccine, along with various doses of beta-glucosylceramide (beta-GC), beta lactosylceramide (beta-LC), or a combination of both (IGL) in different doses. The effect of beta-glycosphingolipids on the immune response towards HBV was tested by fluorescence-activated cell sorting analysis of hepatic and splenic NKT and CD8 lymphocytes, and serum cytokine levels. RESULTS: Beta-sphingolipid treatment altered the hepatic NKT and CD8 lymphocyte distribution. beta-LC, beta GC, and the combination of both augmented anti-HBV immunity, increasing both the anti-HBs titers and the percentage of mice exhibiting high titers. This effect was associated with altered hepatic NKT and CD8+ lymphocyte distribution. CONCLUSIONS: In summary, beta-glycosphingolipids increased the anti-HBV immune response in association with an altered NKT and CD8 lymphocyte distribution, making beta-glycosphingolipids potential potent adjuvants for overcoming non responsiveness to HBV vaccination and augmenting the anti-viral immune response. PMID- 18423948 TI - A vector-based minigene vaccine approach results in strong induction of T-cell responses specific of hepatitis C virus. AB - Multiepitope-based vaccines against hepatitis C virus (HCV) were designed in the form of three minigenes encompassing four domains of the NS3, NS4 and NS5B proteins that contain multiple class I/II restricted epitopes. The polyEp-WT minigene encodes all four domains in fusion, the polyEp-C minigene encodes the same fusion but optimised for mammalian translation and the polyEp-E3 minigene has an additional endoplasmic reticulum targeting sequence. Whereas the minigenes vectorised by DNA were poorly immunogenic, adenovirus vectorisation induced strong and broader IFNgamma-ELISpot and CTL responses in HLA-A2 transgenic mice. In addition, polyEp-WT and polyEp-E3 responses were found cross-reactive in a recombinant Listeria-NS3-based surrogate challenge. This study illustrates the potency of vectorised minigenes in the field of HCV vaccine development. PMID- 18423949 TI - Chimeric vapA/groEL2 DNA vaccines enhance clearance of Rhodococcus equi in aerosol challenged C3H/He mice. AB - Rhodococcus equi remains a significant bacterial pathogen, causing severe pyogranulomatous pneumonia in foals aged 1-3 months. There is no effective vaccine currently available for the prevention of R. equi pneumonia. DNA vaccines are known to offer specific advantages over conventional vaccines. The aim of this study was to demonstrate efficacy of our recombinant DNA vaccine candidates, namely pcDNA3-Re1, pcDNA3-Re3 and pcDNA3-Re5 by combining a heat shock protein GroEL2 to a virulence-associated protein A (VapA) from R. equi to protect C3H/He mice against the R. equi infection. VapA was shown to be strongly recognised by sera from pneumonic foals. All vaccines elicited at least a doubling of the IgG2a/IgG1 ratio in comparison to the controls, indicating a bias to the Th1 response, which is postulated to be crucial for bacterial clearance and protective immunity against intracellular pathogens including R. equi. In addition, the immunised mice showed a significant reduction in R. equi in their lungs at 7 days after the aerosol challenge in comparison to PBS treated mice. However, examination of lung pathology 14 days after the challenge showed no gross differences in pathological changes between the unvaccinated and vaccinated animals. The lack of significant pathological changes suggests that the precise level of protection against R. equi pneumonia in the murine model of infection may not represent a true effectiveness of the potential vaccine candidates, indicating the mouse may not be the ideal non-equine model for vaccine studies and (or) the incomplete immunogenic antigen of vapA-based DNA vaccine constructs that mount an inadequate cell-mediated immune response against the R. equi infection. PMID- 18423950 TI - Immunopathological responses and kinetics of Brucella melitensis Rev 1 infection after subcutaneous or conjunctival vaccination in rams. AB - The innocuousness of the Brucella melitensis Rev 1 live attenuated vaccine strain has never been fully assessed in rams. The immunopathological responses and the kinetics and distribution of the infection induced by this strain were determined after subcutaneous or conjunctival vaccination in both young (3-4 months old) and adult (12 months old) rams. At regular intervals after vaccination the animals were bled for serological studies, and slaughtered for both pathological and bacteriological examinations. The serological response after conjunctival inoculation was of lower intensity and duration than that induced subcutaneously, being the differences more evident in young rams. No genital lesions were produced and genital organs and accessory sexual glands were never found infected, being Rev 1 infection restricted to lymph nodes and spleen. Immunostained Rev 1 bacteria were located intracellularly in plasmablasts, dendritic follicular cells and macrophages in the target lymph nodes, in which cellular hyperplasia was the dominant pathological response. Subcutaneous vaccination induced a generalized infection by 2 weeks after vaccination, being then restricted to the prescapular target lymph node. Infection after conjunctival vaccination was less generalized, being restricted essentially to the cranial lymph nodes. Rev 1 infection was fully cleared by 3 months after vaccination in all animals. These results confirm the innocuousness of B. melitensis Rev 1 vaccine in rams. PMID- 18423951 TI - Behavioural responses to perceived risk of tick-borne encephalitis: vaccination and avoidance in the Baltics and Slovenia. AB - Tick-borne encephalitis (TBE) incidence increased markedly in the Baltics and Slovenia in the early 1990s, but then declined again in some places. Our analyses of temporal and spatial data on TBE incidence and vaccination revealed that over 1970-2005 up-take of vaccination varied in both time and space according to incidence, i.e. was apparently responsive to perceived risk. Since 1999, however, decreases in incidence in many counties within each country have far exceeded vaccination rates or immunity through natural exposure, and in Latvia and Lithuania these changes are correlated with previous incidence. Survey data on human activities in Latvia revealed that people in socio-economic groups whose behaviour put them at highest risk of exposure to ticks in forests, including people with lower education and lowest incomes, are least likely to be vaccinated. We conclude that risk avoidance through changing human behaviour has driven incidence-dependent decreases in TBE infection, but targeted vaccination campaigns could provide more secure protection. PMID- 18423952 TI - Quality of Web-based information on cocaine addiction. AB - OBJECTIVE: To evaluate the quality of web-based information on cocaine use and addiction and to investigate potential content quality indicators. METHODS: Three keywords: cocaine, cocaine addiction and cocaine dependence were entered into two popular World Wide Web search engines. Websites were assessed with a standardized proforma designed to rate sites on the basis of accountability, presentation, interactivity, readability and content quality. "Health on the Net" (HON) quality label, and DISCERN scale scores aiding people without content expertise to assess quality of written health publication were used to verify their efficiency as quality indicators. RESULTS: Of the 120 websites identified, 61 were included. Most were commercial sites. The results of the study indicate low scores on each of the measures including content quality. A global score (the sum of accountability, interactivity, content quality and aesthetic criteria) appeared as a good content quality indicator. CONCLUSIONS: While cocaine education websites for patients are widespread, their global quality is poor. There is a need for better evidence-based information about cocaine use and addiction on the web. PRACTICE IMPLICATIONS: The poor and variable quality of web-based information and its possible impact on physician-patient relationship argue for a serious provider for patient talk about the health information found on Internet. Internet sites could improve their content using the global score as a quality indicator. PMID- 18423953 TI - Mechanical stability of structured bone grafts from the anterior iliac crest. AB - BACKGROUND: The anterior part of the iliac crest is frequently used as a donor site for autogenous bone grafts. Depending on the clinical scenario, the grafts are exposed to compression and shear stresses. Insufficient stability of a bone graft can lead to the collapse of a reconstruction. The object of this study was to quantify the influence of surgery-related parameters like the bony composition (cancellous, unicortical and bicortical), the donor site and the angle of removal on the failure load of the bone plugs (shearing and compression tests). METHODS: A total of 137 bone cylinders varying in bony composition and in their angles to the trajectories in the anterior iliac crest were taken from 48 human hemipelvises with diamond hollow cutters. The failure load was measured in a standardized manner, each bone plug being subjected to two shear tests and three compression tests. Possible differences between groups were evaluated using analysis of covariance techniques with BMI, bone density and cortical thickness as covariates. The statistical significance level was set to 5% (P<0.05), no adjustment for multiple comparisons was done due to the descriptive nature of the study. FINDINGS: Bicortical bone plugs were more resistant than unicortical plugs to both compression and shear stress, and both were more stable than cancellous bone plugs (P<0.05). On average, the compression failure load was about twice as high as the corresponding failure load with shearing tests: the factor was approximately 1.5 for cancellous, approximately 2 for unicortical and approximately 3 for bicortical bone plugs. We could not find a significant effect of the precise donor site and the angle of removal (parallel or perpendicular to the crista) on the mechanical stability of cancellous bone plugs. INTERPRETATION: Surgeons should rather focus on the cancellous or cortical composition of the graft than location of the harvesting site or drilling angle in order to obtain a useful bone plug from the iliac crest. PMID- 18423954 TI - Stress distribution in the intervertebral disc correlates with strength distribution in subdiscal trabecular bone in the porcine lumbar spine. AB - BACKGROUND: It is understood that an interdependence of properties exists between the intervertebral disc and the subdiscal trabecular bone. Determining the biomechanics of this relationship is important in the development of novel spinal implants and instruments. The aim of this study was to analyze this relationship for the porcine lumbar spine and to compare it with that of the human spine. METHODS: The stress distribution within the intervertebral disc of 10 porcine lumbar (L4/L5) motion segments was recorded using a 1.5mm needle pressure transducer. For dynamic loading a specialized testing rig was developed to apply flexion/extension and medial/lateral bending while intervertebral disc stress was simultaneously recorded. The regional variation in mechanical properties of trabecular bone was also examined for an additional 10 porcine (L5) vertebral bodies. For compressive testing of the subdiscal bone, columns were prepared using a low speed cutting saw and subjected to axial compression. FINDINGS: Under pure compressive loading, stress levels within the intervertebral disc were relatively uniform. However, during asymmetric loading large peak stresses were evident in the periphery of the intervertebral disc in areas underlying the annulus fibrosus. The mechanical properties of trabecular bone demonstrated regional variations within the vertebral body. The ratio of compressive yield strength of bone underlying the outer annulus fibrosus to that of bone underlying the nucleus pulposus averaged 1.36. INTERPRETATION: Although the effects of stress distribution and bone mass adaptation cannot be directly related, it is probable that peak stresses arising in the annulus fibrosus during asymmetric loading provide greater stimuli for the underlying bone to undergo adaptive remodeling to withstand the greater forces experienced. Findings of intervertebral stress distribution and strength distribution of subdiscal trabecular bone for the porcine spine may aid in the development of strategies for preclinical animal testing of spinal implants. PMID- 18423955 TI - [Herpetic whitlow: a forgotten occupational hazard]. PMID- 18423956 TI - [Audit of the prehospital management of severe head injured patients in the "Pays de-la-Loire" region]. AB - OBJECTIVE: To study severe head injured patients (SHIP) during prehospital emergency care (Samu) and the first day in the French Pays-de-la-Loire area, in 2005. STUDY DESIGN: Prospective and multicenter. PATIENTS AND METHODS: All patients managed by Samu with GCS or =3.0ng/ml prompted a DRE and a transrectal ultrasound (TRUS) guided, lateralized sextant biopsy. Throughout the three screenings of the ERSPC, Rotterdam, 5040 biopsy sessions were evaluated. MEASUREMENTS: We determined the positive predictive values (PPVs) of a suspicious DRE and normal DRE, which entailed, respectively, the proportion of PCs detected in men with a suspicious DRE or normal DRE divided by, respectively, all biopsied men with a suspicious DRE or normal DRE. RESULTS AND LIMITATIONS: At initial screening, the PPV of a suspicious DRE, in conjunction with an elevated PSA level, to detect PC was 48.6% compared to 22.4% for men with a normal DRE. Both PPVs decreased in consecutive screens: respectively, 29.9% versus 17.1% (screen 2) and 21.2% versus 18.2% (screen 3). Respectively, 71.0% (p<0.001), 68.8% (p<0.001), and 85.7% (p=0.002) of all PCs with a Gleason score >7 were detected in men with a suspicious DRE at screens 1, 2, and 3. A limitation is that only biopsied men were evaluated. CONCLUSIONS: At initial and subsequent screenings, the chance of having cancer at biopsy was higher in men with a suspicious DRE compared to men with a normal DRE (to a lesser extent in subsequent screenings), and the combination of a PSA level > or =3.0ng/ml with a suspicious DRE resulted in detecting significantly more PCs with Gleason score >7. DRE may be useful in more selective screening procedures to decrease unnecessary biopsies and overdiagnosis. PMID- 18423978 TI - Relations between the mitogen-activated protein kinase and the cAMP-dependent protein kinase pathways: comradeship and hostility. AB - Inter- and intracellular communications and responses to environmental changes are pivotal for the orchestrated and harmonious operation of multi-cellular organisms. These well-tuned functions in living organisms are mediated by the action of signal transduction pathways, which are responsible for receiving a signal, transmitting and amplifying it, and eliciting the appropriate cellular responses. Mammalian cells posses numerous signal transduction pathways that, rather than acting in solitude, interconnect with each other, a phenomenon referred to as cross-talk. This allows cells to regulate the distribution, duration, intensity and specificity of the response. The cAMP/cAMP-dependent protein kinase (PKA) pathway and the mitogen-activated protein kinase (MAPK) cascades modulate common processes in the cell and multiple levels of cross-talk between these signalling pathways have been described. The first- and best characterized interconnections are the PKA-dependent inhibition of the MAPKs ERK1/2 mediated by RAF-1, and PKA-induced activation of ERK1/2 interceded through B-RAF. Recently, novel interactions between components of these pathways and new mechanisms for cross-talk have been elucidated. This review discusses both known and novel interactions between compounds of the cAMP/PKA and MAPKs signalling pathways in mammalian cells. PMID- 18423979 TI - Molecular characterisation of growth differentiation factor 9 (gdf9) and bone morphogenetic protein 15 (bmp15) and their patterns of gene expression during the ovarian reproductive cycle in the European sea bass. AB - Members of the transforming growth factor-beta superfamily, growth differentiation factor 9 (GDF9) and bone morphogenetic protein 15 (BMP15), have crucial roles in primary follicle growth in mammals. To initiate investigations into their significance in teleost oogenesis, we set out to clone and characterise the cDNAs of gdf9 and bmp15 and analysed their patterns of gene expression during the ovarian reproductive cycle in the European sea bass (Dicentrachus labrax). Sea bass gdf9 and bmp15 cDNAs were 2200 and 2049 bp long, coding for 438 and 459 amino acids (aas), respectively, and were most similar to zebrafish gdf9 and bmp15 (64.4 and 56.1%, respectively). By Northern analysis, sea bass gdf9 and bmp15 mRNA transcripts were detected in the ovary only of the tissues analysed and their sizes were 2.2 and 2.1 kb, respectively. Dot-blot analysis revealed high levels of gdf9 and bmp15 expression in the ovary during primary oocyte growth and previtellogenesis (July to October), with a significant decline at the onset of vitellogenesis (November) and remaining low until the beginning of new oocyte growth (April/May). There was a highly significant positive correlation (r=0.939) between gdf9 and bmp15 gene expression in individual samples. The high levels of gdf9 and bmp15 mRNA transcripts in the ovary, especially during the previtellogenic growth period suggest an important role for these factors in early primary oocyte growth in the European sea bass. PMID- 18423980 TI - Bilateral ischemic necrosis of the tongue due to disseminated intravascular coagulation. AB - Ischemic necrosis of the tongue is a rare condition because the tongue has a rich blood supply. Temporal arteritis appears to be the most frequent cause of tongue necrosis. The case is presented of an 82-year-old man who developed bilateral ischemic necrosis of the tongue. The necrosis was considered to be a consequence of thrombosis of the bilateral lingual arteries due to disseminated intravascular coagulation. To the authors' knowledge, this is the first reported case of necrosis of the tongue secondary to disseminated intravascular coagulation. PMID- 18423981 TI - Differential impact of TGF-beta and EGF on fibroblast differentiation and invasion reciprocally promotes colon cancer cell invasion. AB - Several studies indicate that cancer-associated fibroblasts play a critical role in cancer cell invasion and metastasis, the hallmarks of malignancy. To better understand the mechanisms underlying such effects, we established a heterotypic model of human fibroblasts (primary colon fibroblasts and immortalized human dermal fibroblasts) in co-culture with human colon cancer cells (HCT-8/E11), using three-dimensional collagen type-I and Matrigel matrices. We report that TGF beta is the unique and dominant factor to provide pro-invasive signals to HCT 8/E11 colon cancer cells from TGF-beta-treated human fibroblasts in three dimensional collagen type I and Matrigel matrices. These effects are not mimicked or reversed by EGF or bFGF, and are associated with the TGF-beta-mediated induction of myofibroblast differentiation and functional markers, such as alpha SMA, the haptotactic matrix molecule TNC, collagen type 1 maturation enzyme P4H, serine protease FAP, and myofibroblast contractility. Accordingly, TGF-beta induced a strong activation of RhoA and stress fiber formation in fibroblasts, with no impact on Rac1-GTP levels. In contrast, EGF down-regulated Rho-GTP levels in fibroblasts, giving permissive signals for Rac1 activation, fibroblast polarization, and invasion. Taken together, our data imply that TGF-beta and EGF exert invasive growth-promoting actions in human colon tumors through a differential and cumulative impact on the stromal and cancer cell compartments. Our data predict that inhibitors directed at this reciprocal molecular and cellular crosstalk will have therapeutic applications for targeting the invasive growth of human primary tumors and their metastatic spread. PMID- 18423982 TI - An acidic extracellular pH induces Src kinase-dependent loss of beta-catenin from the adherens junction. AB - Little attention has been paid to the role of adherens junctions (AJs) in acidic extracellular pH (pHe)-induced cell invasion. Incubation of HepG2 cells in acidic medium (pH 6.6) induced cell dispersion from tight cell clusters, and this change was accompanied by downregulation of beta-catenin at cell junctions and a rapid activation of c-Src. Pretreatment with PP2 prevented the acidic pH-induced downregulation of beta-catenin at AJ and in the membrane fractions. The acidic pHe-induced c-Src activation increased tyrosine phosphorylation of beta-catenin and decreased the amount of beta-catenin-associated E-cadherin. The depletion of membrane-bound beta-catenin coincided with enhanced cell migration and invasion, and this acidic pHe-increased cell migration and invasion was prevented by PP2. In conclusion, this study characterizes a novel signaling pathway responsible for acidic microenvironment-promoted migration and invasive behaviors of cancer cells. PMID- 18423983 TI - Overexpression of AQP5, a putative oncogene, promotes cell growth and transformation. AB - Overexpression of several aquaporins has been reported in different types of human cancer but the role of AQPs in human carcinogenesis has not yet been clearly defined. Here, we demonstrate that ectopic expression of human AQP5 (hAQP5), a water channel expressed in lung, salivary glands, and kidney, induces many phenotypic changes characteristic of transformation both in vitro and in vivo. Furthermore, the cell proliferative ability of AQP5 appears to be dependent upon the phosphorylation of a cAMP-protein kinase (PKA) consensus site located in a cytoplasmic loop of AQP5. In addition, phosphorylation of the PKA consensus site was found to be phosphorylated preferentially in tumors. These findings altogether indicate that hAQP5 plays an important role in human carcinogenesis and, furthermore, provide an attractive therapeutic target. PMID- 18423984 TI - Altered 'three-flash' illusion in response to two light pulses in schizophrenia. AB - BACKGROUND: Disorganization is a core dysfunction in schizophrenia. Coherent thought and behavior rely on the interactive neural responses to temporally discrete external events. Previous studies have demonstrated that a single visual stimulus (event) is abnormally affected by another (as in backward masking), but the integration (or 'synthesis') of temporally discrete events remains largely unexplored in schizophrenia. We examined the perceived interaction of two elementary visual events in schizophrenia patients, using a psychophysical approach. METHODS: Two brief, spatially-coincident foveal light pulses (5 ms) were presented with different inter-stimulus intervals (ISIs). At ISIs around 100 ms, a substantial proportion of the light pulse pairs was paradoxically perceived as three flashes, a known phenomenon in normal subjects. The subjects reported the number of flashes perceived ('one', 'two' or 'three'). RESULTS: Schizophrenia patients (n=28) reported fewer 'three flashes' than normal controls (n=26) at the ISIs where 'three flash' reports were greatest in normal subjects (90 to 110 ms). On the other hand, at longer ISIs (130-310 ms) patients reported 'three flashes' in more trials than did normal subjects. The perception of three flashes in patients was correlated with certain aspects of clinical status, including the positive and general subscales of the PANSS. DISCUSSION: The alteration of the 'three-flash' illusion in schizophrenia suggests that the synthesis of discrete visual events is temporally 'dilated' or distorted, which might contribute to disorganized thought and behavior. PMID- 18423985 TI - Predicting response to atypical antipsychotics based on early response in the treatment of schizophrenia. AB - OBJECTIVE: To test whether early onset of response to antipsychotic medications accurately predicts subsequent response in the treatment of patients with schizophrenia. METHODS: We used data from 5 randomized, double-blind clinical trials comparing olanzapine with other atypical antipsychotic drugs in the treatment of patients with schizophrenia and related disorders, who were at least moderately ill at baseline and who were treated for a minimum of 2 weeks (N=1077). Early response was defined as >or=20% improvement on the PANSS total score at 2 weeks. Conditional probabilities (sensitivity, specificity, positive and negative predictive values) were used to characterize the likelihood of "subsequent response" to treatment (i.e., >or=40% improvement on the PANSS total score with treatment up to 3 months). Subsequent analyses focused on varying thresholds of subsequent response, and at different time points. RESULTS: Most (80%) of subsequent non-responders by 3 months were correctly classified as early non-responders at 2 weeks (high specificity), and 84% of early non-responders at 2 weeks were subsequent non-responders by 3 months (high negative predictive value). For early responders, prediction of subsequent response was substantially lower. A higher threshold (e.g., >or=40% improvement) had greater predictive accuracy at all time points measured. Early non-responders attained less symptom improvement overall, and were more likely to discontinue from treatment. CONCLUSIONS: Early non-response to antipsychotic medications was a robust predictor of subsequent lack of response in the treatment of patients with schizophrenia. Evaluating patients as early as 2 weeks in treatment can help identify non-responders who may benefit from an alternative therapeutic approach. PMID- 18423986 TI - Molecular cloning of Bombyx mori cytochrome P450 gene and its involvement in fluoride resistance. AB - To investigate the effects of fluorosis on development and gene expression profiles of silkworm, highly resistant silkworm strain 441, and highly susceptible silkworm strain 440 were treated with 200ppm fluoride (designated as 440F and 441F) and water (designated as 440DZ and 441DZ). Fluorotic silkworm showed body color and behavior changes. Statistical analysis indicated that growth index of 440F was lower than 440DZ, 441DZ, and 441F. The mortality of 440F was higher than others. Fluorescent differential display enabled us to obtain a differentially expressed cDNA. Bioinformatics analyses indicated that it belonged to cytochrome P450 family, denoted Bmcyp306a1, which contained seven exons and six introns. Phylogenetic tree showed BmCYP306A1 had high homology with Manduca sexta' P450 protein. Expression analysis indicated that Bmcyp306a1 was exclusively expressed in 441DZ and 441F and was down-regulated under fluoride treatment. The tissue-specific expression indicated Bmcyp306a1 had high expression level in midgut and ovary in 441F. The data revealed that there was obvious dose-effect and times relationship with the pathological changes and gene expression. Expression profiles of Bmcyp306a1 suggested that P450 gene was crucial to physiological modification and might be involved in fluoride resistance. PMID- 18423987 TI - Preference of medicine and patient-reported quality of life in community-treated schizophrenic patients receiving aripiprazole vs standard of care: results from the STAR study. AB - OBJECTIVE: To evaluate quality of life and patient preference for schizophrenia treatment in a community based study comparing the use of aripiprazole to the standard of care (SOC). METHOD: This open-label, 26-week, multi-centre, randomised study compared aripiprazole with SOC (olanzapine, quetiapine or risperidone) in patients with schizophrenia (DSM-IV-TR criteria). The primary effectiveness variable was the mean total score of the Investigator Assessment Questionnaire (IAQ) at Week 26. The outcome research variables included the Preference of Medicine (POM) questionnaire, the Quality of Life Scale (QLS), and the EuroQoL-5D (EQ-5D). The results from these outcome research variables are the focus of this paper addressing quality of life and patient preference. RESULTS: A total of 555 patients were randomised to receive aripiprazole (n=284) or SOC (n=271). The OC data at Week 26, reported that more respondents rated the study medication as 'much better' compared with their previous medication in the aripiprazole group versus SOC for patients (59% vs 35%, P<0.001) and caregivers (58% vs 30%, P=0.014). The improvement in QLS total score was also significantly greater in the aripiprazole group compared with SOC--mean change from baseline in QLS total score of 16.21 vs 10.01 (P<0.001) at Week 26 (OC data set). A greater proportion of patients (93% vs 85%; P=0.005) in the aripiprazole group had a satisfactory response on the EQ-5D Self Care Scale; all other EQ-5D scores were similar. CONCLUSION: The study findings suggest that quality of life and patient medication preference measures were better for aripiprazole than for SOC. PMID- 18423988 TI - Structural and functional deficits in human amblyopia. AB - Many neuroimaging tools have been used to assess the site of the cortical deficits in human amblyopia. In this paper, we aimed at detecting the structural and functional deficits in humans with amblyopia, with the aid of anatomic magnetic resonance imaging (aMRI) and functional MRI (fMRI). We designed the visual stimulus to investigate the functional deficits, and delineated the V1/V2 areas by retinotopic mapping. Then we performed the brain parcellation to calculate the volume of the subcortical structure on each individual, and reconstructed the cortical surfaces to measure the cortical thickness. At last, the statistical comparison was carried out to find the structural abnormities and their relationship to the functional deficits. Compared with the normal controls, it is found that the hemisphere difference existed on the unilateral amblyopia subjects, and the functional deficit might come along with the changes in the cortical volume, especially in the occipital lobe. The examined results may provide insight to the study of the neural substrates of amblyopia. PMID- 18423989 TI - Numbness in clinical and experimental pain--a cross-sectional study exploring the mechanisms of reduced tactile function. AB - Pain patients often report distinct numbness of the painful skin although no structural peripheral or central nerve lesion is obvious. In this cross-sectional study we assessed the reduction of tactile function and studied underlying mechanisms in patients with chronic pain and in healthy participants exposed to phasic and tonic experimental nociceptive stimulation. Mechanical detection (MDT) and pain thresholds (MPT) were assessed in the painful area and the non-painful contralateral side in 10 patients with unilateral musculoskeletal pain. Additionally, 10 healthy participants were exposed to nociceptive stimulation applied to the volar forearms (capsaicin; electrical stimulation, twice each). Areas of tactile hypaesthesia and mechanical hyperalgesia were assessed. MDT and MPT were quantified adjacent to the stimulation site. Tactile hypaesthesia in pain patients and in experimental pain (MDT-z-scores: -0.66+/-0.30 and -0.42+/ 0.15, respectively, both p<0.01) was paralleled by mechanical hyperalgesia (MPT-z scores: +0.51+/-0.27, p<0.05; and +0.48+/-0.10, p<0.001). However, hypaesthesia and hyperalgesia were not correlated. Although 9 patients reported numbness, only 3 of them were able to delineate circumscript areas of tactile hypaesthesia. In experimental pain, the area of tactile hypaesthesia could be mapped in 31/40 experiments (78%). Irrespective of the mode of nociceptive stimulation (phasic vs. tonic) tactile hypaesthesia and hyperalgesia developed with a similar time course and disappeared within approximately 1 day. Hypaesthesia (numbness) often encountered in clinical pain can be reproduced by experimental nociceptive stimulation. The time course of effects suggests a mechanism involving central plasticity. PMID- 18423990 TI - Investigation of the role of electrolytes and non-electrolytes on the cloud point and dye solubilization in antidepressant drug imipramine hydrochloride solutions. AB - Antidepressant drug imipramine hydrochloride (IMP) is amphiphilic which shows surfactant-like behavior in aqueous solutions. We have studied the effect of adding electrolytes and non-electrolytes on the micellar behavior of IMP by making cloud point (CP) and dye solubilization measurements. The CP of a 100mM IMP solution (prepared in 10mM sodium phosphate (SP) buffer) was found to decrease with increasing pH, both in the absence as well as presence of added salts. Increase in pH increased the visible absorbance of Sudan III dye solubilized in the drug micelles, implying micellar growth. Addition of increasing amounts of salts to 100mM IMP solutions (at pH 6.7) caused continuous increase in CP due to micellar growth. On the basis of these studies, the binding effect orders of counter- and co-ions have been deduced, respectively, as: Br( )>Cl(-)>F(-) and Li(+)or=3, R<200 km, and 21% at site B. Empirical equations between earthquake magnitude, epicenter distance and precursor time enabled estimation or prediction of an earthquake that will rise at the epicenter distance R from the monitoring site in expecting precursor time T. PMID- 18424053 TI - Tritiation of several antihypertensive agents at high specific activity. AB - Methods are described to synthesize and characterize tritium labeled analogues of telmisartan and lacidipine. PMID- 18424054 TI - [How to take up an expertise for medical liability in anaesthesiology?]. AB - Search for responsibility in medicine became everyday. Anaesthetists are particularly exposed and will be, several times, confronted to it during their career. They have to have knowledge of some necessary elements to get to grips with expertise. Expertise can be asked by a penal jurisdiction. In that case, the anaesthetist can be directly and personally implicationed. When expertise is asked by a civil jurisdiction, it concerns anaesthetists, whichever the (liberal or employee of private). Expertise during administrative procedures concern hospital's anaesthetists. It is important to organize a preparatory meeting in any expertise. Praticians must collect together the complete medical file to establish the most exactly possible, chronology of facts. The anaesthetist can be accompanied by medical consultant appointed by the insurance companies and a lawyer. But he does not have to content with be represented by them. Presence in expertise is essential; praticians can so give evidence of their good faith and answer the expert's questions. Vagueness or doubt are never favorable to pratician. It is also, a responsible and respectful behavior toward the patient. PMID- 18424055 TI - [Three case-reports of viperin envenoming in Morocco]. AB - Snake bites are responsible for a high mortality rate in Africa. Problems for the early care of the victims are many. We published three observations of Moroccan typical viperin severe envenomings (rapidly extensive edema, necrosis, haemorrhagic shock) are presented. The overall mortality of those bites is 4%. In the Maghreb, viperin syndromes are the result of the lebetin viper (Vipera lebetina), the horned viper or sand viper (Cerastes cerastes), sometimes Bitis or Echis sp. Immunotherapy remains effective against haemorrhage, even when administered late, in severe disease (bleeding). Death remains inevitable if antivenin is not being administered or if it is at the stage of multiorgan failure. Heparin is contra-indicated in the acute context, but at distance from the bite, the persistence of inflammatory syndrome can cause phlebitis or pulmonary embolism. Fresh frozen plasma and corticosteroids are ineffective and unnecessary. Signs of gravity are rapidly extensive swelling and systemic manifestations, particularly bleeding. The severity of poisoning is related to the difficulties of access to health centers, the use of traditional medicine for more than half of the victims, the lack of training of caregivers, finally and most importantly, the lack of antivenomous serum,that is not within reach of prescribing at the appropriate places and times. Despite its cost and allergic risk (become rare with the current serums), immunotherapy which is the only weapon effective against the venom, should be part of emergency essential drugs. PMID- 18424056 TI - Postsynaptic density protein PSD-95 expression in Alzheimer's disease and okadaic acid induced neuritic retraction. AB - In order to understand how plasticity is related to neurodegeneration, we studied synaptic proteins with quantitative immunohistochemistry in the entorhinal cortex from Alzheimer patients and age-matched controls. We observed a significant decrease in presynaptic synaptophysin and an increase in postsynaptic density protein PSD-95, positively correlated with beta amyloid and phosphorylated Tau proteins in Alzheimer cases. Furthermore, Alzheimer-like neuritic retraction was generated in okadaic acid (OA) treated SH-SY5Y neuroblastoma cells with no decrease in PSD-95 expression. However, in a SH-SY5Y clone with decreased expression of transcription regulator LMO4 (as observed in Alzheimer's disease) and increased neuritic length, PSD-95 expression was enhanced but did not change with OA treatment. Therefore, increased PSD-95 immunoreactivity in the entorhinal cortex might result from compensatory mechanisms, as in the SH-SY5Y clone, whereas increased Alzheimer-like Tau phosphorylation is not related to PSD-95 expression, as suggested by the OA-treated cell models. PMID- 18424058 TI - Effect of a chronic GSM 900 MHz exposure on glia in the rat brain. AB - Extension of the mobile phone technology raises concern about the health effects of 900 MHz microwaves on the central nervous system (CNS). In this study we measured GFAP expression using immunocytochemistry method, to evaluate glial evolution 10 days after a chronic exposure (5 days a week for 24 weeks) to GSM signal for 45 min/day at a brain-averaged specific absorption rate (SAR)=1.5 W/kg and for 15 min/day at a SAR=6 W/kg in the following rat brain areas: prefrontal cortex (PfCx), caudate putamen (Cpu), lateral globus pallidus of striatum (LGP), dentate gyrus of hippocampus (DG) and cerebellum cortex (CCx). In comparison to sham or cage control animals, rats exposed to chronic GSM signal at 6 W/kg have increased GFAP stained surface areas in the brain (p<0.05). But the chronic exposure to GSM at 1.5 W/kg did not increase GFAP expression. Our results indicated that chronic exposure to GSM 900 MHz microwaves (SAR=6 W/kg) may induce persistent astroglia activation in the rat brain (sign of a potential gliosis). PMID- 18424057 TI - Oxidative injury and neuropathy in diabetes and impaired glucose tolerance. AB - Clinical studies suggest that impaired glucose tolerance (IGT) is associated with the development of neuropathy. The aim of the current study was to determine if neuropathy developed in the female Zucker Diabetic Fatty (ZDF) rat, an animal model of IGT and type 2 diabetes. The ZDF rat develops impaired glucose tolerance (IGT) when fed a control diet, and frank diabetes when fed a high fat diet. Following 10 weeks of hyperglycemia, sensory nerve action potentials (SNAP) and compound motor action potentials (CMAP) were reduced and sensory conduction velocities were slowed (distal>proximal) in the tail and hind limb in ZDF animals with IGT and frank diabetes (p<0.01). Neuropathy was coupled with evidence of increased reactive oxygen species (ROS) and cellular injury in dorsal root ganglion (DRG) neurons from IGT animals. Our study supports the hypothesis that neuropathy develops in an animal model of IGT and is associated with evidence of oxidative injury in DRG and peripheral nerves. PMID- 18424059 TI - Histological and histomorphometric analyses of calcium phosphate cement in rabbit calvaria. AB - PURPOSE: To assess bone regeneration in critical sized defects in the rabbit calvarium, filled with the bone substitute calcium phosphate cement. MATERIAL AND METHODS: Circular bone defects (8mm) were made in both parietal bones of 10 rabbits. One of the defects was filled with the calcium phosphate cement, and the other received autogenous bone harvested from the calvaria. The animals were killed at 3 or 6 weeks (n=5). Data analysis included qualitative assessment of the calvarial specimens and histomorphometric analysis was used to quantify the amount of new bone within the defects. RESULTS: The microscopic analysis of the samples showed bone healing with both calcium phosphate cement and autogenous bone graft. Data obtained from the histomorphometric analysis were statistically analyzed using 2-way analysis of variance and the Tukey's test. Data analysis showed that the autogenous bone graft had significantly more new bone compared with calcium phosphate cement at 3 and 6 weeks. Calcium phosphate cement at 6 weeks presented similar results to autogenous bone at 3 weeks. Both treatments presented an increase in bone healing with time. CONCLUSION: Treatments allowed bone regeneration that increased with time, however surgical cavities treated with the autogenous graft had more bone formation than those with calcium phosphate cement. PMID- 18424060 TI - Distraction osteogenesis in secondary microsurgical mandible reconstruction: report of seven cases. AB - INTRODUCTION: Fibula, ilium and scapula are the preferred donor areas for vascularised bone grafts. For secondary mandibular reconstruction, however, compromises often have to be made as a result of the poor quality of the covering soft tissue and recipient vessels especially in irradiated patients. Callus distraction osteogenesis is a complementary method for optimising the intermaxillary relationship and the facial profile from an aesthetic point of view. MATERIALS AND METHODS: Thirteen distractions were performed in seven patients following microsurgical mandibular reconstruction over a period of 4 years, all by the same surgeon. Intra-oral distraction devices were predominantly applied. After a 7-day healing period, the callus distraction was begun, involving two advisements per day of 0.5mm each. The distraction devices were removed after a consolidation phase of 4-5 months. RESULTS: Ten of the 13 distractions were uneventful with an average bone lengthening of 20mm, retained after a follow-up time of 6-47 months (average 27 months). The intermaxillary relationship and the facial profile were improved in all cases. Distraction plate fractures, screw loosening or absence of new bone formation were the complications encountered in the other three patients. CONCLUSION: Regarding the high complication rate, a successful outcome cannot be expected with greater certainty. Thus the indication should be stringent when including this procedure within the possible treatment alternatives available to the clinician. PMID- 18424061 TI - [Cardiac tamponade as an initial manifestation of systemic lupus erythematosus]. PMID- 18424062 TI - Quality of life and anxiety and depressive disorder comorbidity. AB - The present investigation evaluated the relations among anxiety and depressive disorder comorbidity and quality of life (QOL) by utilizing self-report measures of life satisfaction and functional disability. Participants were 94 individuals who were presented for treatment at an outpatient anxiety disorders clinic and 26 nonclinical participants. Results indicated that participants diagnosed with anxiety disorders reported lower QOL than did nonclinical participants. Anxiety disorder comorbidity did not additionally impact QOL; however, presence of a depressive disorder comorbid with an anxiety disorder did negatively impact QOL as these individuals reported significantly more functional disability and less life satisfaction than did individuals with anxiety disorders alone or those without a psychiatric diagnosis. These results highlight the negative nature of anxiety disorders and improve clarification on the role of diagnostic comorbidity on QOL among those with an anxiety disorder. PMID- 18424063 TI - Psychometric properties of the Trauma Relevant Assumptions Scale. AB - This article describes the psychometric properties of a novel questionnaire, i.e. the Trauma Relevant Assumptions Scale (TRAS). The added value of the TRAS over previous trauma relevant belief questionnaires is that the TRAS enables measuring valence and rigidity of beliefs simultaneously. Both aspects are thought to be predictive of the development of chronic PTSD symptoms. For the exploratory factor analysis, the TRAS was administered to 309 adult volunteers. Principal components analysis yielded two factors: Assumptions about Self and Assumptions about the World. The two-factor structure was confirmed in a sample of 185 traumatized individuals. The TRAS seems to be a valid and reliable instrument, which is strongly related to post-trauma symptoms and has good discriminative validity. Apart from research settings, the TRAS may also be suitable in therapeutic settings to identify the severity of dysfunctional assumptions, and to assess the progress in change from negative assumptions to more positive assumptions. PMID- 18424064 TI - No improvement in neurocognitive outcomes after off-pump versus on-pump coronary revascularisation: a meta-analysis. AB - The popularity of off-pump (beating heart) coronary artery bypass grafting (CABG) was initially stimulated by numerous theoretical benefits including lower incidence of stroke and neurocognitive dysfunction. With a postoperative stroke rate of less than 1% for elective CABG, it has been very difficult to demonstrate any significant differences in this outcome between techniques. However, changes in neurocognitive function are more common in the postoperative setting and thus provide greater power for demonstrating improvement with changes in surgical technique. The aim of this meta-analysis was to assess whether there were significant differences in neurocognitive outcomes in patients after undergoing off-pump versus on-pump CABG. A database search for prospective randomised controlled trials of off-pump versus on-pump CABG in any language was conducted. Eight trials incorporating 892 patients fulfilled all the inclusion criteria for reporting of neurocognitive outcomes, and were able to be included in this meta analysis. Sufficient data were available across the seven studies to combine results for five neurocognitive tests (Rey Auditory Verbal Learning, Grooved Pegboard, Trail A and B, and Digit Symbol). Overall there were no convincing differences in outcomes in neurocognitive testing between off-pump and on-pump CABG groups. The results of this meta-analysis show that there are no significant neurocognitive benefits when comparing off-pump versus on-pump CABG. PMID- 18424065 TI - Retrograde type A dissection after endovascular stenting of the descending thoracic aorta. Is the risk real? AB - OBJECTIVE: Retrograde type A dissection during or after endoluminal graft repair of the descending thoracic aorta is a potentially lethal complication unique to thoracic endografting. Our aim is to increase its awareness and to review possible etiological factors. METHODS: Two hundred and eighty-seven patients with different thoracic aortic pathologies were treated with endovascular prostheses over the last 6 years (February 2000 to March 2006) under a single-site protocol. A retrospective review was conducted to identify any retrograde aortic dissections by both chart and film review. Factors that may have contributed to its formation were also documented. This population was analyzed for the complication of retrograde aortic dissection as well as the factors related to its occurrence. RESULTS: Seven patients (2.4%) with a gender distribution of three males and four females experienced a retrograde type A dissection within this sample at a median of 202 days. The mean age was 74 years (range 53-83). Aortic pathologies included aortic dissections (n=6) and thoracic aortic aneurysm (n=1). There were (n=3) 43% retrograde type A dissections identified within the perioperative period. Balloon angioplasty was performed in 71.4% (n=5). Two female patients (28.6%) had this event identified within their initial hospitalization with fatal consequences. Overall mortality was 57% (n=4) with extension of dissection the primary cause of death n=3 and open surgical repair (n=1) after an extension of retrograde dissection. CONCLUSIONS: Female gender, use of stent-grafts for dissection and possible aggressive balloon angioplasty may play a role in the cause of retrograde type A dissection. A close surveillance program is recommended when using thoracic endografts outside the recommended device instructions for use. PMID- 18424066 TI - The outcome of pregnancy following topiramate treatment: a study on 52 pregnancies. AB - In spite of a substantial increase in the use of topiramate at child bearing age, very little is known regarding its use in pregnancy. We describe the outcome of 52 pregnancies with 41 liveborn infants from which it seems that topiramate reduces birth weight without decreasing gestational age at delivery, but does not seem to increase the risk for structural defects. There was an increased rate of spontaneous abortions not related to the drug effects. PMID- 18424067 TI - The potential role of histone deacetylase inhibitors in the treatment of non small-cell lung cancer. AB - Non-small-cell lung cancer (NSCLC) arises from a complex series of genetic and epigenetic changes leading to uncontrolled cell growth and metastases. The exponential growth in the level of research about the histone deacetylase (HDAC) enzymes, responsible for deacetylating core nucleosomal histones and other proteins, has been driven by the ability of HDAC inhibitors to modulate transcriptional activity. As a result, this therapeutic class is able to block angiogenesis and cell cycling, and promote apoptosis and differentiation. The mechanisms resulting in the antiproliferative biologic effects of these agents are not yet known. Clinical experience indicates these agents generally well tolerated, and active in several haematological and solid tumours. HDAC inhibitors, under clinical evaluation in the treatment of NSCLC patients, are pivanex, CI-994, vorinostat, and LBH589. Here, we discuss about the potential role of HDAC inhibitors focusing on their activity, tolerability, efficacy and future development, in the treatment of NSCLC. PMID- 18424068 TI - Detection and quantification of human and bovine noroviruses by a TaqMan RT-PCR assay with a control for inhibition. AB - Noroviruses are single-stranded RNA viruses belonging to the family Caliciviridae. They are a major cause of epidemic and sporadic gastroenteritis in humans and calves. Reverse transcription-polymerase chain reaction (RT-PCR) has become the "gold standard" for detection of noroviruses in faecal and environmental samples. However, false negative results due to co-concentration of RT-PCR inhibitors are a continuous concern. A TaqMan real-time RT-PCR assay making use of a foreign internal RNA control and a RNA standard was developed. Very interestingly, this method is capable of detecting human noroviruses belonging to genogroups I and II, and bovine noroviruses belonging to genogroup III. Inhibitors were removed efficiently by 1/10 dilution of the sample or addition of bovine serum albumin to the RT-PCR mix. This assay was validated with human and bovine stool samples previously tested for norovirus by conventional RT PCR. The ability to detect norovirus in stool samples that were negative by conventional RT-PCR assay demonstrate the higher sensitivity of the TaqMan assay compared to the conventional RT-PCR assay. This real-time RT-PCR assay allows the detection of both human and bovine noroviruses, avoids false negative results and is able to quantify the level of norovirus contamination. PMID- 18424069 TI - Cyclosporin A inhibits the production of IL-17 by memory Th17 cells from healthy individuals and patients with rheumatoid arthritis. AB - Recent evidence from several studies indicated that IL-17-producing Th17 cells can represent the key effector cells in the induction and development of autoimmune disorders. Cyclosporine A (CsA) is a commonly used immunosuppressant to treat lots of autoimmune diseases including rheumatoid arthritis (RA). Here, we demonstrated that PBMCs and purified CD4(+) T cells from healthy individuals and patients with RA could be induced to produce large amounts of IL-17 after stimulation with anti-CD3 plus anti-CD28 mAbs. Phenotypic analysis indicated that the majority of IL-17-producing cells were Th17 cells with memory phenotype. The addition of CsA into cell cultures significantly inhibited the IL-17 production by Th17 cells at protein and at mRNA levels. Compared to the PBMCs from normal individuals, PBMCs from the patients with RA produced higher levels of IL-17 that was also significantly inhibited by CsA both at protein and at mRNA levels. The mechanism might be the effect of CsA on the T cells activation because the expression of CD69 and CD25 molecules on T cells was markedly reduced in the presence of CsA. Taken together, these results demonstrated that CsA suppressed the IL-17 production and inhibited the Th17 cells differentiation from both healthy individuals and patients with RA. PMID- 18424070 TI - Heme content of recombinant catalase from Psychrobacter sp. T-3 altered by host Escherichia coli cell growth conditions. AB - The catalase gene of Psychrobacter sp. T-3 was cloned, and the gene product (PktA) was overexpressed in Escherichia coli. The specific activity of the purified PktA was slightly lower than that of the native purified enzyme obtained from Psychrobacter sp. T-3. Spectrophotometric measurements of the purified enzymes suggested that the recombinant PktA contains a mixture of heme b and d, although the native enzyme contains the sole heme b. An addition of the heme precursor 5-aminolevulinic acid (ALA) to the medium increased the heme b content of the recombinant PktA, and the resulting enzyme showed higher specific activity than the native enzyme. This is the first report that shows the heme content of overproduced catalase altered by the host cell growth conditions. PMID- 18424071 TI - G-protein-dependent and -independent pathways regulate proteinase-activated receptor-2 mediated p65 NFkappaB serine 536 phosphorylation in human keratinocytes. AB - The mechanisms underpinning the coupling of GPCRs, such as PAR-2, to the phosphorylation of p65 NFkappaB have not been investigated. In the current study we found that trypsin and the selective PAR-2 activating peptide, 2f-LIGKV-OH, stimulated large and sustained increases in the serine 536 phosphorylation of p65/RelA in a transfected skin epithelial cell line and primary keratinocytes. Parallel experiments showed that in both cell types, p65 NFkappaB phosphorylation is mediated through the selective activation of IKK2. Treatment with PKC inhibitor GF109203X or PKCalpha siRNA reduced phosphorylation at 15 min but not 30 min, whilst rottlerin, a selective PKCdelta inhibitor and PKCdelta siRNA reduced the response at both time points. Pre-treatment of cells with the novel Gq/11 inhibitor YM-254890 and Gq/11 siRNA caused a similar pattern of inhibition and also reduced PAR-2-mediated NFkappaB transcriptional activity. Furthermore, stimulation of cells through a novel PAR-2 mutant PAR-2(34-43), delayed p65 phosphorylation but was without effect on the kinetics of ERK activation. Inhibition of Gi or G12/13 pathways by pertussis toxin pre-treatment or over expression of the RGS mutant Lsc, also did not effect NFkappaB phosphorylation. Taken together these data indicate dependency for Gq/11 in early phosphorylation of p65 NFkappaB and this subsequently affects initial NFkappaB-dependent gene transcriptional activity, however later regulation of p65 is unaffected. Overall these novel data demonstrate an IKK2-dependent, predominantly G-protein independent pathway involved in PAR-2 regulation of NFkappaB phosphorylation in keratinocytes. PMID- 18424072 TI - A PAK4-LIMK1 pathway drives prostate cancer cell migration downstream of HGF. AB - Hepatocyte growth factor (HGF) is associated with tumour progression and increases the invasiveness of prostate carcinoma cells. Cell migration and invasion requires reorganisation of the actin cytoskeleton; processes mediated by the Rho family GTPases. p21 activated kinase 4 (PAK4), an effector of the Rho family protein Cdc42, is activated downstream of HGF. We report here the novel finding that in prostate cancer cells PAK4 binds to and phosphorylates LIM kinase 1 (LIMK1) in an HGF-dependent manner. We show for the first time that variations in the level of PAK4 expression change the level of cofilin phosphorylation in cells, a change we correlate with LIMK1 activity, cell morphology and migratory behaviour. We identify for the first time a direct and localised interaction between PAK4 and LIMK1 within cells using FRET: FLIM. Moreover we show here that HGF mediates this interaction which is concentrated in small foci at the cell periphery. PAK4 and LIMK1 act synergistically to increase cell migration speed, whilst a reduction in PAK4 expression decreases cell speed. It is well established that unphosphorylated (active) cofilin is a required to drive cell migration. Our results support a model whereby HGF-stimulated cell migration also requires a cofilin phosphorylation step that is mediated by PAK4. PMID- 18424073 TI - A profile of Fritiof S. Sjostrand--the founding editor. AB - The Journal of Ultrastructure Research was founded in 1957 by Fritiof S. Sjostrand, who served as Editor-in-Chief until 1990, when the journal changed the name to the Journal of Structural Biology. This profile summarizes the developments that led to the start of the journal and aspects of Fritiof Sjostrand's scientific and personal carrier. PMID- 18424074 TI - Intracellular precipitation of hydroxyapatite mineral and implications for pathologic calcification. AB - In contrast to physiologic biomineralization occurring in bones, teeth and otoconia in vertebrates, calcification of soft tissues occurs in many pathologic conditions. Although similarities have been noted between the two processes, and despite the important clinical consequences of ectopic calcification, the molecular mechanisms regulating ectopic calcification are poorly understood. Although calcification is mainly extracellular, intracellular calcification has been reported and might indeed contribute to pathologic calcification of soft tissues. To better understand the process of intracellular calcification as a potential origin for pathologic calcification, and to examine the role of proteoglycans in this process, we investigated a pattern of intracellular nucleation and growth of hydroxyapatite in Madin-Darby Canine Kidney (MDCK) epithelial cells using electron microscopy, secondary ion mass spectroscopy (NanoSIMS), cytochemical staining, immunolabeling and biochemical analysis. We report here that under mineralizing cell culture conditions where beta glycerophosphate (betaGP) was added as an exogenous organic source of phosphate, betaGP-cleaving alkaline phosphatase activity increased and hydroxyapatite crystals subsequently nucleated within intracellular, membrane-bounded compartments. The small, leucine-rich proteoglycan decorin was also upregulated and associated with mineral in these cultures. Such information provides insight into the mechanisms leading to pathologic calcification and describes a process whereby hydroxyapatite deposition in cells might lead to ectopic calcification. PMID- 18424075 TI - Long-term outcome of gamma-knife surgery in temporal lobe epilepsy. AB - PURPOSE: Indication of gamma-knife surgery (GKS) in temporal lobe epilepsy (TLE) remains a matter of debate. Either positive or negative results have been reported in studies with a maximum follow-up of 24 months. No long-term data have been published yet. METHODS: We collected data from 15 TLE patients who underwent GKS and whose follow-up was longer than 24 months. Localisation and extension of the epileptogenic zone (EZ) were assessed by SEEG in eight patients. Ten patients were presenting with mesial TLE (mTLE) while in the other five SEEG showed that the EZ was extending beyond mesio-temporal structures. GKS procedure was targeted to the EZ at a dose of 21.1+/-2.6Gy at the margin. RESULTS: The mean duration of the follow-up was 60+/-22.3 months. At last follow-up, seven patients (46.7%) were free of disabling seizure. The last seizure following GKS occurred on average after a delay of 19.75+/-20.62 months. No patient suffered recurrent seizures after a long period free of disabling seizures. No significant prognostic predictor could be individualized in this population. However, 6 of the 10 patients with mTLE (60%) were seizure free whereas only 1 of the 5 (20%) whose EZ was more extended exhibited the same seizure outcome. CONCLUSION: When a positive outcome is achieved, the risk of seizure recurrence remains low at long term. In patients with typical mTLE, long-term GKS results may be closed to those observed after conventional surgery. Conversely, this procedure should not be proposed to patients in whom SEEG results suggest that the EZ is not restricted to mesial temporal structures. PMID- 18424076 TI - Thrombocytopenia during levetiracetam therapy. AB - A 64-year-old patient with symptomatic epilepsy developed thrombocytopenia during treatment with levetiracetam (LEV). As no other medical reason could be evaluated, a medication side effect was postulated. The only new drugs were valproic acid (since 3 weeks) and levetiracetam (since 3 days). After valproic acid medication was ended, thrombocytopenia did not improve and even worsened further. Finally levetiracetam administration was ended and trombocytopenia resolved rapidly and completely within few days. PMID- 18424077 TI - Angiotensin II enhances EGF receptor expression levels via ROS formation in HaCaT cells. AB - BACKGROUND: Recent work has shown a novel function of angiotensin II (Ang II) in skin wound healing in which reactive oxygen species might be involved. As Ang II is known to increase superoxide production by activating NADPH oxidase in some non-phagocytic cells, we hypothesized that the produced superoxide by NADPH activation could contribute to the regulation of epidermal growth factor receptor (EGFR) in keratinocytes. OBJECTIVE: We examined whether Ang II could generate superoxide and enhance EGFR expression levels in HaCaT cells. METHODS: Superoxide formation was assessed by using hydroethidine. EGFR expression levels were examined by Western blotting. RESULTS: Ang II (1-100 microM) increased the superoxide formation. Ang II (1-100 microM) resulted in a dose-dependent increase in cell proliferation in HaCaT cells. Heparin-binding epidermal growth factor activated the EGFR at 5-10 min. Although Ang II did not activate the EGFR, the expression levels of EGFR protein were increased in HaCaT cells treated with Ang II (1 microM) at 6h. Apocynin, a NADPH oxidase inhibitor, decreased the expression levels of EGFR. Xanthine/xanthine oxidase system, an exogenous superoxide generating system, enhanced the EGFR protein expression. Although Ang II did not affect the nitric oxide (NO) production, a NO synthase inhibitor N(omega)-nitro-l-arginine methyl ester suppressed the Ang II-induced EGFR expression levels in HaCaT cells. Thus, constitutive NO is required for the Ang II-induced EGFR expression in HaCaT cells. CONCLUSION: These results suggest that Ang II enhances the cell proliferation and EGFR expression via superoxide production under the regulation of NO in HaCaT cells, implying that Ang II may regulate the proliferation, differentiation and tumorigenesis of the epidermis by harmonizing the superoxide and NO production. PMID- 18424078 TI - Intense pulsed light modulates the expressions of MMP-2, MMP-14 and TIMP-2 in skin dermal fibroblasts cultured within contracted collagen lattices. PMID- 18424079 TI - Role of the interleukin 15 96516A>T and IL15 96330C>A gene polymorphisms in Caucasian patients with chronic plaque psoriasis. PMID- 18424080 TI - The "sense of agency" and its underlying cognitive and neural mechanisms. AB - The sense of agency is a central aspect of human self-consciousness and refers to the experience of oneself as the agent of one's own actions. Several different cognitive theories on the sense of agency have been proposed implying divergent empirical approaches and results, especially with respect to neural correlates. A multifactorial and multilevel model of the sense of agency may provide the most constructive framework for integrating divergent theories and findings, meeting the complex nature of this intriguing phenomenon. PMID- 18424082 TI - Corpus callosum damage in heavy marijuana use: preliminary evidence from diffusion tensor tractography and tract-based spatial statistics. AB - Heavy marijuana use has well established long term consequences for cognition and mental health, but the effect on brain structure is less well understood. We used an MRI technique that is sensitive to the structural integrity of brain tissue combined with a white matter mapping tractography technique to investigate structural changes in the corpus callosum (CC). Diffusion tensor imaging (DTI) was obtained in eleven heavy marijuana users who started using marijuana in early adolescence and eleven age matched controls. Mean diffusivity (MD) and fractional anisotropy (FA) (which measure structural integrity and tract coherence, respectively) were analysed within the corpus callosum which was spatially defined using tractography and tract-based spatial statistics (TBSS). MD was significantly increased in marijuana users relative to controls in the region of the CC where white matter passes between the prefrontal lobes. This observation suggests impaired structural integrity affecting the fibre tracts of the CC and is in keeping with previous reports of altered and diversified activation patterns in marijuana users. There was a trend towards a positive correlation between MD and length of use suggesting the possibility of a cumulative effect of marijuana over time and that a younger age at onset of use may predispose individuals to structural white matter damage. Structural abnormalities revealed in the CC may underlie cognitive and behavioural consequences of long term heavy marijuana use. PMID- 18424081 TI - Two components of the Myb complex, DMyb and Mip130, are specifically associated with euchromatin and degraded during prometaphase throughout development. AB - The Drosophila Myb protein, DMyb, is a transcription factor important for cell proliferation and development. Unlike the mRNAs produced by mammalian myb genes, Drosophila myb transcripts do not fluctuate substantially during the cell cycle. A comprehensive analysis of the localization and degradation of the DMyb protein has now revealed that DMyb is present in nuclei during S phase of all mitotically active tissues throughout embryogenesis and larval development. However, DMyb and Mip130, another member of the Myb complex, are not uniformly distributed throughout the nucleus. Instead, both proteins, which colocalize, appear to be specifically excluded from heterochromatic regions of chromosomes. Furthermore, DMyb and Mip130 are unstable proteins that are degraded during prometaphase of mitosis. The timing of their degradation is reminiscent of Cyclin A, but at least for DMyb, the mechanism differs; although DMyb degradation is dependent on core APC/C components, it does not depend on the Fizzy or Fizzy-related adaptor proteins. DMyb levels are also high in actively endoreplicating polyploid cells, but there is no indication of cyclical degradation. We conclude that cell cycle specific degradation of DMyb and Mip130 is likely to be utilized as a key regulatory mechanism in down-regulating their levels and the activity of the Myb complex. PMID- 18424083 TI - Attentional bias of competitive interactions in neuronal networks of early visual processing in the human brain. AB - Multiple objects in a visual scene compete for neuronal representation. We investigated competitive neuronal dynamics in cortical networks of early visual processing in the human brain. Coloured picture streams flickered at 7.42 Hz, evoking the steady-state visual evoked potential (SSVEP), an electrophysiological response of neuronal populations in early visual areas synchronised by the external pacemaker. While these picture streams were at a fixed location in the upper left and right quadrant, respectively, additional competing picture streams flickering at a different frequency were continuously changing the distance to the stationary streams by slow motion. Analysis of the 7.42 Hz SSVEP amplitude revealed significant amplitude decreases when the competing stimulus was closer than about 4.5 degrees of visual angle. Sources of the SSVEP suppression effect were found in early visual areas of the ventral and dorsal processing streams. Attending the stationary stimulus resulted in no difference in 7.42 Hz SSVEP amplitude regardless of spatial separation to the competing stimulus. Contrary to the predictions of the model, we found co-amplification of the competing stimulus at close spatial proximity accompanied by an increase of an intermodulation frequency, suggesting integrated neuronal processing of target and competing stimuli when both streams are close together. PMID- 18424084 TI - Template-O-Matic: a toolbox for creating customized pediatric templates. AB - Processing pediatric neuroimaging data is a challenge due to pervasive morphological changes that occur in the human brain during normal development. This is of special relevance when reference data is used as part of the processing approach, as in spatial normalization and tissue segmentation. Current approaches construct reference data (templates) by averaging brain images from a control group of subjects, or by creating custom templates from the group under study. In this technical note, we describe a new, and generalized method of constructing such appropriate reference data by statistically analyzing a large sample (n=404) of healthy children, as acquired during the NIH MRI study of normal brain development. After eliminating non-contributing demographic variables, we modeled the effects of age (first, second, and third-order terms) and gender, for each voxel in gray matter and white matter. By appropriate weighting with the parameter estimates from these analyses, complete tissue maps can be generated automatically from this database to match a pediatric population selected for study. The algorithm is implemented in the form of a toolbox for the SPM5 image data processing suite, which we term Template-O-Matic. We compare the performance of this approach with the current method of template generation and discuss the implications of our approach. PMID- 18424085 TI - High-resolution study of nuclear magnetic relaxation dispersion of purine nucleotides: effects of spin-spin coupling. AB - By combining magnetic field cycling in the range from 0.1mT to 7T with high resolution NMR detection the T(1) relaxation dispersion (nuclear magnetic relaxation dispersion (NMRD)) of protons in the nucleotides adenosine mono phosphate and guanosine mono-phosphate was measured in a site-specific way. While at high field the individual spins have distinctly different T(1) times, their scalar spin-spin interaction fulfills at low field the condition of strong coupling and leads to convergence of their T(1) dispersion curves. In addition, the spin-spin coupling can lead to oscillatory components in the relaxation kinetics traceable to a coupling between spin polarization and coherence in the relaxation process. As a consequence the NMRD curves do not directly reflect the spectral density function of the motional processes, but the effects of motion and spin coupling must be separated for a reliable evaluation. A theoretical approach is described allowing such an analysis. PMID- 18424086 TI - [Prevalence of reading disabilities in early elementary school: impact of socioeconomic environment on reading development in 3 different educational zones]. AB - OBJECTIVE: Socioeconomic status (SES) has a known influence on academic achievement. Most studies, however, were conducted in English-speaking countries. Because recent cross-linguistic studies suggest that reading English is much harder to learn than reading other languages, an epidemiological study was conducted in French investigating the impact of socioeconomic background on early reading development. DESIGN: One thousand and twenty second-grade children (476 girls and 544 boys) from 20 different schools participated in the study. Approximately 1/3 of the children lived and were schooled in a high SES area, 1/3 in an intermediate SES area, and one final third in a very low SES area. Assessment of reading, writing and mathematical skills was conducted initially in small groups. Children with suspected learning difficulties were further tested individually. Forty-two children of equivalent age who repeated the first grade received similar individual testing. RESULTS: Average reading scores were in accordance with chronological age, without gender differences. Children from low SES schools had academic performances significantly lower than their peers. Boys exhibited superior arithmetic skills than girls. A significant reading delay was observed in 12.7% of children. The prevalence of poor reading was highly correlated with the area of schooling, varying from 3.3% in the high SES area to 24.2% in low SES area. CONCLUSION: The high rate of children from our sample with a significant delay in reading depended on general socioeconomic environment. An understanding of the origin of such differences is mandatory for defining and coordinating preventive actions and appropriate interventions. PMID- 18424087 TI - WITHDRAWN: Molecular phylogeny of the Papuan microhylid frogs (Anura: Microhylidae: Asterophryinae and Genyophryninae) inferred from 12S and 16S mitochondrial DNA sequences. AB - This article has been withdrawn consistent with Elsevier Policy on Article Withdrawal (http://www.elsevier.com/locate/withdrawalpolicy). The Publisher apologizes for any inconvenience this may cause. PMID- 18424089 TI - Phylogenies without roots? A plea for the use of vouchers in molecular phylogenetic studies. PMID- 18424088 TI - Phylogeography of the Pygmy Rain Frog (Pristimantis ridens) across the lowland wet forests of isthmian Central America. AB - We used a phylogeographic approach to elucidate the evolutionary history of a lineage of frogs, known as Pristimantis (formerly Eleutherodactylus) ridens (Anura: Brachycephalidae), restricted to the wet forests occurring along the Caribbean versant of isthmian Central America as well as the disjunct wet forest on the Pacific slope of Costa Rica. We placed our phylogeographic study of P. ridens within a larger molecular phylogenetic analysis of Central American Pristimantis. All phylogenetic inferences were based on a 1455 base pair fragment of mitochondrial DNA, containing the complete ND2 gene and five flanking tRNA genes. Our reconstruction of the intraspecific phylogeny of P. ridens yielded a basal trichotomy dating to an estimated 12+ million years ago (Ma), consisting of central Panama, western Panama, and Costa Rica plus Honduras. Thus, the presence of P. ridens appears to predate the completion of the Isthmus 3.1Ma. Using a parametric bootstrap (SOWH) test, we evaluated four a priori zoogeographic hypotheses for the origin and spread of P. ridens. This analysis suggested that the P. ridens populations on the Caribbean versant of Costa Rica were established by Pacific versant ancestors only recently, in contrast to the very old lineages found in Panama. Our results support a model of Miocene colonization, long-term geographic stasis, followed by rapid dispersal across the Caribbean lowlands during the Pliocene or Pleistocene. PMID- 18424091 TI - Malpositioning of radial head prostheses: an in vitro study. AB - Nonanatomic insertion of radial head prostheses may result in maltracking and capitellar erosion. The purpose of this study was to analyze the ability to perform anatomic radial head replacement and to study radiocapitellar prosthetic subluxation under unstable conditions. In vitro measurements were performed in 10 intact elbows and after insertion of radial head prostheses (rigid uncemented, rigid cemented, bipolar). The diameter and length of the proximal radius were approximately restored. However, prosthesis-shaft malalignment, up to 21 degrees , was observed frequently. A posterolateral rotatory stress after resection of 50% of the coronoid caused a mean radiocapitellar subluxation of more than 30% for the rigid prostheses, whereas bipolar devices self-aligned with a mean subluxation similar to intact elbows. In conclusion, it was not possible to consistently insert radial head prostheses anatomically. Marked radiocapitellar subluxation occurred for the rigid but not for the bipolar implants. Clinical trials are needed to analyze these findings. PMID- 18424092 TI - Reconstruction of posttraumatic bone defects of the humerus with vascularized fibular graft. AB - Humeral nonunions still present a challenge to the orthopedic surgeon. Many methods of treating recalcitrant, posttraumatic humeral shaft nonunions have been described, with varying degrees of success. The present report reviews our experience with the use of vascularized fibular grafting for the treatment of large humeral defects. We treated 13 patients, with an average length of the humeral defect of 10.5 cm. Nine patients healed primarily, 3 required additional bone grafting, and 1 had a second fibular transplant. The mean period to radiographic bone union was 6 months. Only 5 patients regained full range of motion of the shoulder and elbow. The vascularized fibular graft is a reliable reconstructive procedure for recalcitrant pseudoarthrosis of the humerus in which the bony gap is greater than 6 to 7 cm, especially when traditional procedures have not provided the expected result. PMID- 18424093 TI - The relationship between the pressure adjacent to the ulnar nerve and the disease causing cubital tunnel syndrome. AB - We investigated the relationship between cubital tunnel pressure in patients with cubital tunnel syndrome with osteoarthritis and those without osteoarthritis. We studied 31 elbows in 29 patients. We divided the patients into two groups: one associated with osteoarthritis and the other not associated with osteoarthritis. In the latter group, there was ulnar nerve subluxation in 10 elbows and cubitus valgus in 3. Cubital tunnel pressure was measured intraoperatively with a fiberoptic microtransducer. The extraneural pressure with the elbow flexed was significantly increased in patients with osteoarthritis and those without osteoarthritis. The pressure within the cubital tunnel in osteoarthritic elbows was significantly higher than that in those without osteoarthritis. Moreover, the pressure of osteoarthritic elbows significantly increased from proximally to distally within the cubital tunnel, whereas the pressure in elbows without osteoarthritis was high only proximally. Thus, cubital tunnel pressure could be a more important causative factor for cubital tunnel syndrome in the elbows with osteoarthritis than in those without osteoarthritis. PMID- 18424094 TI - The determination of stratum corneum thickness: an alternative approach. AB - The individual thickness of the stratum corneum is required to normalise drug permeation profiles in dermato-pharmacokinetic studies. The thickness is often estimated using tape-stripping combined with transepidermal water loss measurements. A linear transformation of Fick's first law is used to relate the progressively thinner barrier with the corresponding increase in transepidermal water loss and to estimate the thickness by linear regression. However, the data from an important subset of subjects are poorly fitted to this linear model. This is typically due to the removal of loose outer layers of stratum corneum, which do not contribute significantly to barrier function. This work proposes two alternative non-linear models. All three models were used to fit data from 31 in vivo tape-stripping experiments and their outcomes and goodness-of-fit compared. The results suggest that the linear model may overestimate the stratum corneum thickness and is open to subjectivity regarding the selection of data points to be fitted. The non-linear models satisfactorily fitted all the data, including all data points. No significant differences were found between the thicknesses derived from the two non-linear models. However, the analysis of the goodness-of fit of the models to the data suggests a preference for a baseline-corrected approach. PMID- 18424095 TI - Leaky enteric coating on ranitidine hydrochloride beads: dissolution and prediction of plasma data. AB - The present research is based on the hypothesis that leaky enteric-coated pellets formulations are able to provide sustained input for drugs that have an absorption window, such as ranitidine hydrochloride, without jeopardizing their bioavailability. Leaky enteric-coated pellets formulations are defined as enteric coated pellets that allow some of the drug to be released from the formulation in gastric fluid. Different approaches to making leaky enteric-coated pellets were investigated using extrusion-spheronization followed by spray coating. Leaky enteric coats were formulated using a commonly used enteric polymer, Eudragit L 30 D-55, combined with soluble compounds including lactose, PEG 8000 and surfactants (Span 60 (hydrophobic) or Tween 80 (hydrophilic)). The rate of drug release from the formulations in simulated gastric fluid can be tailored by varying the additive's amount or type. All leaky enteric-coated formulations studied completely released the drugs within 30 min after changing dissolution medium to phosphate buffer, pH 6. Predictions of plasma concentration-time profiles of the model drug ranitidine hydrochloride from leaky enteric-coated pellets in fasted conditions and from immediate-release formulations were performed using computer simulations. Simulation results are consistent with a hypothesis that leaky enteric-coated pellets formulations provide sustained input for drugs shown to have an absorption window without decreasing bioavailability. The sustained input results from the combined effects of the formulation and GI transit effects on pellets. The present research demonstrates a new application of knowledge about gastrointestinal transit effects on drug formulations. It also shows that enteric-coating polymers have new applications in areas other than the usual enteric-coated formulations. The hypothesis that a leaky enteric-coated pellets formulation may maintain or increase the bioavailability of drugs that have a window of absorption is still to be confirmed by further in vivo studies. PMID- 18424096 TI - Adiponectin receptor R1 is upregulated by valproic acid but not by topiramate in human hepatoma cell line, HepG2. AB - Valproic acid (VPA) is an effective and widely used anticonvulsant, associated with metabolic adverse effects such as weight gain, hyperinsulinemia, hyperleptinemia and hypoadiponectinemia. The aim of this study was to evaluate the influence of VPA and topiramate (TPM) on adiponectin binding receptors, adipoR1 and adipoR2, in human liver cancer cells, HepG2. AdipoR1 but not adipoR2 gene expression was upregulated by VPA treatment. TPM did neither affect adipoR1 nor adipoR2 gene expression. Given the tight association between VPA treatment, metabolic side effects and the adipocytokine-axis, upregulation of adipoR1 possibly represents a favoured and insulin-sensitizing mechanism. PMID- 18424098 TI - Cancer chemopreventive activity of "rosin" constituents of Pinus spez. and their derivatives in two-stage mouse skin carcinogenesis test. AB - Natural resin acids present in rosin of Pinus spez., including isopimaric acid (1), mercusic acid (2), neoabietic acid (3), dehydroabietic acid (4), and podocarpic acid (8), as well as resin acid derivatives 8beta,9alpha,13alpha-H tetrahydroabietic acid (5), 8alpha,9alpha,13alpha-H-tetrahydroabietic acid (6), 13alpha-H-Delta(8)-dihydroabietic acid (7), maleopimaric acid (9), and fumaropimaric acid (10), were studied for their possible inhibitory effects on Epstein-Barr virus early antigen (EBV-EA) activation induced by 12-O tetradecanoylphorbol-13-acetate (TPA). Compounds 1, 3, 4, 7, and 10 (IC(50): 352, 330, 311, 340, and 349, respectively) exhibited strong inhibitory effects compared to the other compounds. Among these, 1, 4, and 7 were selected to examine their effects on in vivo two-stage mouse skin carcinogenesis induced by 7,12-dimethylbenz[a]anthracene (DMBA) as initiator and TPA as promoter. Treatment with compounds 4 and 7 (85 nmol) along with DMBA/TPA inhibited papilloma formation up to week 8 and the percentage of papilloma bearers in these two groups was approximately 80% at week 20. The average number of papillomas formed per mouse was 4.4 and 4.2 even at week 20 (p>0.05). Compounds 4 and 7 exhibited high activity in the in vivo anti-tumor-promoting test. In addition, rosin was examined in vivo for its chemopreventive effect. Treatment with rosin (50 MUmol) along with DMBA (100 MUg)/TPA (1 MUg) inhibited papilloma formation up to week 8 and the percentage of papilloma bearers in this group was less than 80% at week 20. The average number of papillomas formed per mouse in the rosin-treated group was 3.8 even at week 20 (p>0.05). The in vivo two-stage mouse skin carcinogenesis test revealed that rosin possessed a pronounced anticarcinogenetic effect, and its high activity is due to the synergism of the diterpenes contained in it. PMID- 18424099 TI - Artichoke leaf extract (Cynara scolymus) reduces plasma cholesterol in otherwise healthy hypercholesterolemic adults: a randomized, double blind placebo controlled trial. AB - Cardiovascular diseases are the chief causes of death in the UK, and are associated with high circulating levels of total cholesterol in the plasma. Artichoke leaf extracts (ALEs) have been reported to reduce plasma lipids levels, including total cholesterol, although high quality data is lacking. The objective of this trial was to assess the effect of ALE on plasma lipid levels and general well-being in otherwise healthy adults with mild to moderate hypercholesterolemia. 131 adults were screened for total plasma cholesterol in the range 6.0-8.0 mmol/l, with 75 suitable volunteers randomised onto the trial. Volunteers consumed 1280 mg of a standardised ALE, or matched placebo, daily for 12 weeks. Plasma total cholesterol decreased in the treatment group by an average of 4.2% (from 7.16 (SD 0.62) mmol/l to 6.86 (SD 0.68) mmol/l) and increased in the control group by an average of 1.9% (6.90 (SD 0.49) mmol/l to 7.03 (0.61) mmol/l), the difference between groups being statistically significant (p=0.025). No significant differences between groups were observed for LDL cholesterol, HDL cholesterol or triglyceride levels. General well-being improved significantly in both the treatment (11%) and control groups (9%) with no significant differences between groups. In conclusion, ALE consumption resulted in a modest but favourable statistically significant difference in total cholesterol after 12 weeks. In comparison with a previous trial, it is suggested that the apparent positive health status of the study population may have contributed to the modesty of the observed response. PMID- 18424100 TI - Anti-inflammatory and analgesic effects and molecular mechanisms of JCICM-6, a purified extract derived from an anti-arthritic Chinese herbal formula. AB - The anti-inflammatory and anti-nociceptive effects and the molecular mechanisms of JCICM-6, a purified extract derived from an anti-arthritic Chinese herbal formula composed of Caulis Sinomenii, Aconiti laterralis Preparata, Rhizoma Curcumae longae, Radix Paeoniae albae, and Cortex Moutan, were examined for the first time. JCICM-6 was prepared using pharmaceutical extraction technology, purified by Amberlite XAD-7HP polymeric resin. Pharmacologically, in carrageenan induced edema and carrageenan-evoked thermal hyperalgesia in paws of rats, the oral administration of JCICM-6 at dosages of 0.4, 0.8, and 1.6g/kg demonstrated significant inhibition with a dose-dependent manner. Mechanistic studies showed that JCICM-6 effectively decreased the production of the pro-inflammatory cytokines of IL-6 and IL-1beta and expression of COX-2 and iNOS proteins, and simultaneously elevated the level of anti-inflammatory cytokine IL-4 in the carrageenan-injected rat paw tissues and exudates. The positive reference drug, indomethacin at a dosage of 10mg/kg, demonstrated inhibitory potency in both rat models, but it could not augment the production of IL-4, indicating JCICM-6 and indomethacin might possess different pharmacological properties and molecular mechanisms although both have anti-inflammatory and analgesic effects in rats. These results suggest that JCICM-6 would be a valuable candidate for further investigation as a new anti-arthritic drug. PMID- 18424101 TI - Tissue distribution and excretion of resveratrol in rat after oral administration of Polygonum cuspidatum extract (PCE). AB - PURPOSE: Polygonum cuspidatum extract as a traditional Chinese medicine is extracted from the dried rhizome and root of Polygonum cuspidatum Sieb.et Zucc. Resveratrol is one of its active components. Studies were performed in rats to define the tissue distribution and excretion of resveratrol in urine and bile, and to characterize (if possible) any metabolites of resveratrol observed in tissues after ig 20mg/kg Polygonum cuspidatum extract. METHOD: For tissue distribution studies, tissues (300 mg) were homogenized and centrifuged with methanol, and metabolites found in selected tissue extract were identified by LC/MS/MS. For urinary and biliary excretion experiments, urine and bile samples were cleaned up by using solid-phase extraction (SPE) with polyamide cartridges. All the concentrations of resveratrol in these biological samples were determined by HPLC with UV detection. RESULT: After a single oral dose of 20mg/kg PCE in rats, resveratrol was mainly distributed in stomach, duodenum, liver and kidney with detectable metabolites resveratrol monoglucuronide and resveratrol monosulfate. The majority of the resveratrol was excreted as metabolites, only 0.59% and 0.027% of the dosage were excreted in urine and bile respectively as unchanged drug within 24h. PMID- 18424102 TI - Inhibiting enoyl-ACP reductase (FabI) across pathogenic microorganisms by linear sesquiterpene lactones from Anthemis auriculata. AB - Enoyl-ACP reductase (FabI) is a key enzyme of the type II fatty acid biosynthesis (FAS-II) pathway and a validated antimicrobial target. In the current study, three linear sesquiterpene lactones obtained from Anthemis auriculata, namely anthecotulide (1), 4-hydroxyanthecotulide (2) and 4-acetoxyanthecotulide (3) were evaluated for specific inhibitory effects against the FabI enzyme from three pathogenic microorganisms, Plasmodium falciparum (PfFabI), Mycobacterium tuberculosis (MtFabI) and Escherichia coli (EcFabI). In addition, the compounds were also tested against two elongation enzymes from the plasmodial FAS-II system, beta-ketoacyl-ACP reductase (PfFabG) and beta-hydroxyacyl-ACP deydratase (PfFabZ). The compounds showed clear differentiation in inhibition of FabI enzymes from different microorganisms. Anthecotulide (1) was most active against MtFabI (IC(50) 4.5 microg/ml), whereas the oxygenated derivatives thereof (compounds 2 and 3) specifically inhibited plasmodial FAS-II enzymes, PfFabI and PfFabG (IC(50) values 20-75 microg/ml). All compounds were inactive towards EcFabI. In whole cell assays, all three compounds exhibited antimalarial and antibacterial activities. PMID- 18424103 TI - Effect of Echinacea augustifolia extract on cell viability and differentiation in mammary epithelial cells. AB - Echinacea spp. are popularly used as an herbal medicine or food supplement for enhancing the immune system and activating biological property in different tissues. In this study we show the biological effect of Echinacea augustifolia extract on cell viability and cell differentiation in mammary epithelial cell lines. These effects have been observed in two different cell line derived from mouse (HC11) and bovine (BME-UV). Echinacea extract enhanced cell liability from 100 to 1000 ng/ml in association with growth factors, epidermal growth factor (EGF) or insulin, but also without EGF (p<0.05) up to 37% vs. control. This effect may be modulated by MAPK and Akt activation that Echinacea extract treatment increased and/or by a reduction of caspase 3 activity, showed a dose response decrease after Echinacea treatment. Finally Echinacea extract was able to increase (p<0.05) at 100 ng/ml beta-casein expression in association with PRL (5 microg/ml). These data demonstrate that Echinacea angustifolia extract can stimulate mammary epithelial cell physiology and may be considered a candidate to support mammary gland activity during a mammogenetic and lactogenetic state. PMID- 18424104 TI - Effect of two artichoke extracts (36_U and 36_EB) on rat ileum (with respect to bowel syndrome) and the peristaltic threshold. AB - The diagnosis and treatment of irritable bowel syndrome (IBS) are complicated. Artichoke extracts are well known to be helpful in various gastrointestinal disorders. A hydrophilic extract 36_U mainly containing luteolin-7-glycoside, luteolin-7-O-glucoside, small amounts of cynarin and luteolin increased contraction of rat ileum. This is mainly mediated by 5-HT(3) - and 5-HT(2) receptors but not 5-HT(4) receptors as can be derived by using specific antagonists such as tropisetrone, GR113806 and ketanserine. Additional mechanisms (receptors) are involved since the combination of these three antagonists was not able to fully prevent the contractive effect of extract 36_U. The lipophilic extract 36_EB mainly containing cynarin, luteolin including its glycosides, and cholorogenic acid in contrast to extract 36_U had a relaxing effect which could hardly be washed out. It was diminishing a serotonin effect and was not modified by ACh or substance P. The peristaltic threshold, i.e. the distension necessary for inducing a pathophysiologically relevant propulsion activity, is one of the important features being correlated with IBS. The peristaltic threshold was decreased by both serotonin and extract U_36. From the data it can be derived that the extract 36_U may be useful in IBS combined with obstipation when gastrointestinal contraction is necessary, whereas 36_EB may be useful in IBS combined with diarrhea when gastrointestinal relaxation is desired. Especially interesting are the influence on the threshold. It would be interesting to know which effects are mediated via cynarin and luteolin or its glycosides. PMID- 18424105 TI - Radioprotective effects of propolis and quercetin in gamma-irradiated mice evaluated by the alkaline comet assay. AB - The radioprotective effects of ethanolic extract of propolis (EEP) and quercetin on the white blood cells of the whole-body irradiated CBA mice were investigated. Irradiation was performed using a gamma-ray source ((60)Co), and absorbed dose was 9 Gy. The efficiency of test components was evaluated when given intraperitoneally (ip) at a dose of 100 mg kg(-1) for 3 consecutive days before and/or after irradiation. Moreover, possible genotoxic effects of test components were also assessed on non-irradiated animals. For each experimental group leukocyte count was determined and the primary DNA damage in leukocytes was assessed using the alkaline comet assay. The higher efficiency of EEP and quercetin was observed when given preventively. The results suggest that propolis and quercetin given to mice before irradiation protect their white blood cells from lethal effects of irradiation and diminish primary DNA damage as confirmed by the alkaline comet assay. Positive results obtained on gamma-irradiated mice given EEP and quercetin, complementary with our earlier observations on survival of irradiated mice, indicate that these compounds could be considered effective non-toxic radioprotectors. The exact mechanisms of radioprotection by these compounds and their effects on DNA repair processes are still to be elucidated. PMID- 18424106 TI - Suppressive effects of bee venom on the immune responses in collagen-induced arthritis in rats. AB - The effect of bee venom (BVA) on the development of type II collagen (CII) induced arthritis (CIA) in rats has been studied. Male rats were immunized with an emulsion of 200 microg of CII and complete Freund's adjuvant (CFA). The rats were then given intraperitoneally (i.p.) injection of a suspension of BVA or saline during the experiment. The effect of BVA on cellular responses to CII was examined. In the control rats, the onset of arthritis was observed at the 24th day after the CII-immunization, and the severity of CIA was developed gradually. As compared with rats treated with saline, BVA i.p. injected at doses of more than 20 microl/100g mouse once a day for 14 days inhibited the ability of inguinal lymph node cells to produce T cell cytokines interleukin-1beta, -2, -6, tumor necrosis factor-alpha and interferon-gamma when the cells were obtained from rats 24 days after immunization and cultured in vitro with CII. When rats were injected i.p. with sheep red blood cells, hemagglutination titers in BVA treated and control rats did not differ significantly when low doses of BVA was given to rats. However, i.p. injection of BVA at doses of more than 10 microl/100g/day suppressed antibody production. Pretreatment of rats with BVA could inhibit the development of collagen arthritis even when 10-20 microl/100g/day of the BVA were used for pretreatment. Interestingly, higher doses than 10 microlBVA/100g mouse were much effective for arthritis incidence. Treatment of rats with BVA prevented the development of collagen arthritis in a dose-dependent manner. Doses of BVA (15 and 20 microl/100g) resulted in decreased incidence of arthritis. In conclusion, therapeutic i.p injection with BVA improved the clinical course of the disease and the immune response to CII. PMID- 18424107 TI - Fully automatic quantification of knee osteoarthritis severity on plain radiographs. AB - OBJECTIVE: Although knee osteoarthritis (OA) is a major public health issue causing chronic disability, there is no objective or accurate method for measurement of the structural severity in general clinical practice. Here we have established a fully automatic program KOACAD (knee OA computer-aided diagnosis) to quantify the major OA parameters on plain knee radiographs, validated the reproducibility and reliability, and investigated the association of the parameters with knee pain. METHODS: KOACAD was programmed to measure joint space narrowing at medial and lateral sides, osteophyte formation, and joint angulation. Anteroposterior radiographs of 1979 knees of a large-scale cohort population were analyzed by KOACAD and conventional categorical grading systems. RESULTS: KOACAD automatically measured all parameters in less than 1s without intra- or interobserver variability. All parameters, especially medial joint space narrowing, were significantly correlated with the conventional gradings. In the parameters, osteophyte formation was associated with none of the joint space parameters, suggesting different etiologic mechanisms between them. Multivariate logistic regression analysis after adjustment for age and confounding factors revealed that medial joint space narrowing and varus angulation of knee joints were risk factors for the presence of pain (594/1979 knees), while neither lateral joint space nor osteophyte area was. CONCLUSION: KOACAD was shown to be useful for objective, accurate, simple and easy evaluation of the radiographic knee OA severity in daily clinical practice. This system may also serve as a surrogate measure for the development of disease-modifying drugs for OA, just as bone mineral density does in osteoporosis. PMID- 18424108 TI - The osteoarthritis initiative (OAI) magnetic resonance imaging quality assurance methods and results. AB - OBJECTIVE: To outline the osteoarthritis initiative's (OAI) magnetic resonance (MR) system quality assurance (QA) processes and present the first 3 years' results. METHOD: OAI MR QA included acquisitions evaluated manually at each of the four sites and other acquisitions assessed by an automated computer program. Key image characteristics such as signal-to-noise, contrast-to-noise, signal uniformity, T2 relaxation times, local and global geometric distortion were quantified monthly using the automated program. RESULTS: Uniformly high quality, artifact-free subject images were obtained from all four OAI 3 tesla MR facilities. Over a 3-year period, key criteria for quantitative cartilage morphometry were excellent with a 190.0 mm diameter and 148.0 mm length object having reproducible diameter (0.04% RMS CV) and length (0.56% RMS CV). This resulted in spherical volume reproducibility of 0.46% RMS CV. Ghost levels were consistently <0.2%. T2 relaxation time varied longitudinally site-by-site from 2.3 to 18.8% RMS CV. All other measures of MR system stability were met except: 3.0 mm and 5.0 mm slice thicknesses were consistently larger than expected; knee coil signal uniformity and signal level varied significantly over time. CONCLUSIONS: OAI MR QA results compared favorably to prior publications and identified similar technical issues for geometric measurements. The longitudinal variations measured in the OAI QA process should have minimal impact on the accuracy and reproducibility of cartilage thickness and volume quantification. This stability should enable direct comparison of baseline and follow-up images. Cross-comparison of the results from all four OAI sites reveals that the MR systems are sufficiently uniform to enable results to be combined. PMID- 18424109 TI - Notes on a cautionary (tall) tale about respondent-driven sampling: a critique of Scott's ethnography. AB - Serious methodological and ethical flaws are detailed in an ethnographic study of a respondent-driven sampling (RDS) project for drug users in Chicago. The study is also disconnected from the larger social context within which the project operated, and from the existing literature on human-subject problems the author claims he "discovered" about RDS - problems common to traditional outreach projects that researchers have known about and managed successfully for years. Due to an admitted bias in the author's sampling, and an eagerness to accept respondents' claims uncritically, the author's results are not generalizeable to RDS projects operating in other cities, in Chicago itself, or even to the specific project studied. PMID- 18424110 TI - Adrift from the moorings of good public policy: ignoring evidence and human rights. PMID- 18424111 TI - Law enforcement and Australia's 2001 heroin shortage: evaluating the evidence. PMID- 18424112 TI - Going soft on evidence and due process: Canada adopts US style harm maximization. AB - After recently adopting a US style drug policy, the Canadian government rejected the recommendations of an independent review of Vancouver's Safer Injecting Facility and ignored the compelling supportive evidence in apparent readiness to close the centre. PMID- 18424113 TI - The implicit rules of evidence-based drug policy: a U.S. perspective. PMID- 18424114 TI - Decline in needle exchange use in Vancouver, Canada coincides with the Australian heroin shortage. PMID- 18424115 TI - Sinus augmentation using human mesenchymal stem cells loaded into a beta tricalcium phosphate/hydroxyapatite scaffold. AB - OBJECTIVE: Implant placement in the posterior maxilla may often be contraindicated because of insufficient bone volume and presence of the maxillary sinus. In these situations, sinus floor augmentation frequently has been proposed as the best treatment. This clinical study was based on the hypothesis that the clinical effectiveness of adult mesenchymal stem cells (MSCs) loaded to the biphasic scaffold. METHODS: In this report, the clinical and radiographic results are presented on 6 consecutively treated patients using MSCs in combination with biphasic hydroxyl apatite/ beta-tricalcium phosphate (HA/TCP) for sinus elevation. All the patients in the study had less than 3 mm initial bone height in the posterior maxillary area (IBH). MSCs were cultured and expanded from bone marrow aspirate for each patient. Three months after sinus elevation, radiographic evaluation was performed for the patients and the secondary bone height was measured (SBH(1)). In the second stage surgery, 30 implants were placed. Trephine bur was used as a pilot drill and a core biopsy was obtained from each implant site. Prosthetic rehabilitation of the patients was performed after 4 months. Secondary bone height was measured 9 months after implant placement (SBH(2)). RESULTS: Of 30 implants, 28 (93%) were considered clinically successful. Two implants were removed due to mobility at the time of surgical exposure. Histologic evaluation of the biopsy specimens revealed numerous areas of osteoid and bone formation HA/TCP, with no evidence of inflammatory cell infiltrate. Mean bone regenerate was 41.34%. Clinically, no complications were observed, and all implants were considered clinically osseointegrated after 4 months. Mean bone height was measured 3 and 12 months after sinus grafting (mean of SBH(1)= 12.08 mm and mean of SBH(2)= 10.08 mm). CONCLUSIONS: These clinical and histological findings suggest that sinus grafting with HA/TCP in combination with MSCs provide a viable therapeutic alternative for implant placement. The findings suggest that the addition of MSCs to bone derivative/substitute materials may enhance bone formation in the maxillary sinus area. Of course more studies with the control groups are needed for the evaluation of this method as a clinical solution for the patients. PMID- 18424116 TI - Congenital epulis: prenatal ultrasonographic and postnatal MR features with pathologic correlation. AB - On prenatal ultrasonogram, a fetus was found to have a protruding mass in the oral cavity. Postnatal magnetic resonance imaging (MRI) revealed the mass to have 2 components: a main portion exhibiting isointensity on T1- and T2-weighted images with mild contrast enhancement and a contrast-enhancing rim showing T1 isointensity and T2 hyperintensity. Histologic examination revealed a congenital granular cell tumor with prominent peripheral vessels. PMID- 18424117 TI - Influence of shaft design on the shaping ability of 3 nickel-titanium rotary systems by means of spiral computerized tomography. AB - OBJECTIVE: To evaluate the influence of shaft design on the shaping ability of 3 rotary nickel-titanium (NiTi) systems. STUDY DESIGN: Sixty curved mesial canals of mandibular molars were used. Specimens were scanned by spiral tomography before and after canal preparation using ProTaper, ProFile, and ProSystem GT rotary instruments. One-millimeter-thick slices were scanned from the apical end point to the pulp chamber. The cross-sectional images from the slices taken earlier and after canal preparation at the apical, coronal, and midroot levels were compared. RESULTS: The mean working time was 137.22 +/- 5.15 s. Mean transportation, mean centering ratio, and percentage of area increase were 0.022 +/- 0.131 mm, 0.21 +/- 0.11, and 76.90 +/- 42.27%, respectively, with no statistical differences (P> .05). CONCLUSIONS: All instruments were able to shape curved mesial canals in mandibular molars to size 30 without significant errors. The differences in shaft designs seemed not to affect their shaping capabilities. PMID- 18424118 TI - Efficacy of intraosseous injections of anesthetic in children and adolescents. AB - OBJECTIVE: The goal of this study was to determine the efficacy of the intraosseous (IO) injections of anesthetic as a primary technique in children and adolescents. STUDY DESIGN: A cohort of 181 children and adolescents underwent a total of 225 sessions of IO injections of 4% articaine with 1:200,000 epinephrine using the Quick Sleeper 2 system. RESULTS: Evaluations could be performed in 215 sessions (171 patients, 247 teeth), yielding success rates of 91.2% (sessions) and 91.9% (teeth). The success rate was 95% (133 of 140) for temporary teeth (endodontics 96.6%, restorations 100%, extractions 88%) and 87.9% (94 of 107) for permanent teeth (endodontics 92.3%, restorations 89.9%, extractions 75%). No difference was noted in terms of age (P > .05). No cases of biting of mucosa or postinjection pain were noted. CONCLUSIONS: The IO injection of anesthetic using a computer-controlled osseous perforation and delivery system can be considered as a good alternative or supplement to classic infiltration techniques in children and adolescents. PMID- 18424119 TI - Influence of CBCT exposure conditions on radiation dose. AB - BACKGROUND: Cone-beam computed tomography (CBCT) has been changing the way dental practitioners use imaging. The radiation dose to the patient and how to effectively reduce the dose is still not completely clear to most users of this technology. OBJECTIVE: The objective of this study was to quantitate the change in radiation dose when using different CBCT settings. METHODS: A CBCT machine was modified to allow different setting combinations. The variables consisted of 4 different mA choices (2, 5, 10, and 15), 2 kVp choices (100 and 120), and 3 fields of view (6 inches, 9 inches, and 12 inches). A radiation phantom with 10 thermoluminescent dosimeters (TLD) was used to measure radiation dose. One specific setting (15 mA, 120 kVp, and 12-inch FOV) was scanned 3 times to determine consistency. RESULTS: The CBCT showed less than 5% variance in radiation dose values. An overall reduction in dose of about 0.62 times was achieved by reducing the kVp from 120 to 100. When reducing the field size the dose decreased 5% to 10%, while for organs that escaped the direct beam the reduction was far greater. CONCLUSIONS: A reduction in radiation dose can be achieved by using the lowest exposure settings and narrow collimation. PMID- 18424120 TI - Mesenchymal stem cells enhance bone regeneration in rat calvarial critical size defects more than platelete-rich plasma. AB - OBJECTIVE: The objective of this study was to compare culture-expanded bone marrow-derived mesenchymal stem cell and PRP loaded to natural bone mineral (Bio Oss) and beta-TCP for rat calvarial bone repair. METHODS: Twenty-two adult male Sprague-Dawley rats were randomly divided into 2 groups (n = 11). In the first group, 2 calvarial defects, 5 mm in diameter, were prepared in the parietal bone. The left defect was filled with Bio-Oss plus PRP while the right defect was filled with mesenchymal stem cell cultured on Bio-Oss. Kasios as a bone substitute was replaced in the second group. The animals were humanely killed 6 weeks postsurgery and the amount of the bone regeneration evaluated using histometric analysis. RESULT: The bone fill length in the calvarial defect had statistically significant difference with other groups (P < .05). In both groups, mesenchymal stem cell revealed a higher percentage of bone formation in comparison with the PRP groups. CONCLUSIONS: New synthetic bone substitutes may offer a better condition for bone regeneration compared to the traditional bone substitute in combination with mesenchymal stem cells. They remained in the defect and contributed bone regeneration. The use of the mesenchymal stem cell in conjunction with bone substitutes can enhance bone regeneration more than PRP. PMID- 18424122 TI - Orbital floor reconstruction: a retrospective study of 21 cases. AB - OBJECTIVE: The purpose of this retrospective study was to investigate the diagnostic methods, therapeutic principles, surgical approach, and materials used for orbital floor reconstruction. STUDY DESIGN: This study consisted of 21 cases with orbital fractures treated at the Hospital of Stomatology, Sichuan University, China, between July 2002 and June 2006. Inclusion criteria were patients with fractures of the orbital floor with bone defects. Patients were retrospectively analyzed for gender, age, mechanism of injury, classification of fracture, and complications. RESULTS: CT scans were 100% accurate in diagnosing the fractures. Shaped autogenous bone, titanium mesh, and Medpor were respectively implanted under the periosteum of the orbital floor in 5 cases, 10 cases, and 6 cases. All the patients had good results including significant improvements in appearance and function after surgery. There were no severe permanent complications. Two cases had postoperative wound infections, and 1 case had temporary blindness that resolved completely. CONCLUSIONS: CT scan is the first choice of investigation for an orbital floor fractures. The objectives of treatment for an orbital floor fracture with a bony defect are reduction of the prolapsed orbital contents and reconstruction of the orbital floor with repair materials, to restore the normal orbital floor and orbital capacity. A subciliary incision was adopted in our surgery. At present, porous polyethylene and titanium mesh are considered to be the ideal orbital floor repair materials. Titanium mesh was used in fractures with large defects that were not easy to fix without obvious enophthalmos. Porous polyethylene can be used in fractures when there is a need to restore the orbital volume. PMID- 18424123 TI - Arthrocentesis of the temporomandibular joint: a proposal for a single-needle technique. AB - Arthrocentesis is a method of flushing out the temporomandibular joint (TMJ) that is currently performed by providing a double access to the joint space. Several studies have shown that arthrocentesis of the upper compartment of the TMJ may be a highly effective method to restore normal maximal mouth opening and functioning. Nonetheless, the classical 2-needle technique has some limits, such as the low tolerability and difficulty in performing it in the presence of intra articular adherences. The adoption of a single-needle for both fluid injection and aspiration might have some advantages with respect to the traditional 2 needle approach in terms of time of execution, tolerability, and retention of medication. These potential advantages have to be assessed with future randomized and controlled clinical trials. PMID- 18424124 TI - Removal of dissolved organic matter and phthalic acid esters from landfill leachate through a complexation-flocculation process. AB - Dissolved organic macromolecules (DOM) and micropollutants, such as phthalic acid esters (PAEs), are often enriched in landfill leachate. PAEs were detected in 10 leachate samples from three landfills in Hubei province, central China. In particular, di-(2-ethyhexyl) phthalate (DEHP) concentration was higher than 70microgL(-1) in most samples, and the highest DEHP concentration was in the colloid fraction (1000Da-0.45microm). Use of flocculants may result in flocculation and precipitation of colloids and associated contaminants, such as DEHP. Biodegradation is not effective for long chain PAEs or the landfill leachate, given the low ratio of BOD5/COD (less than 0.2) and deficient phosphorus content. Complexation-flocculation is therefore proposed as an alternative process for removing DOM and PAEs. Jar-test experiments were employed to investigate the mechanism and feasibility of the process. Among tested coagulants (ferric chloride, aluminum sulfate and poly aluminum chloride (PAC)), PAC was the most efficient. Hydrophobic contaminants with log KOC greater than 4 and DOM in wastewater can be removed simultaneously through the complexation flocculation process. PMID- 18424125 TI - Real-time optical detection of methicillin-resistant Staphylococcus aureus using lytic phage probes. AB - Staphylococcus aureus (S. aureus)-specific bacteriophage was used as a probe for detection of methicillin-resistant S. aureus (MRSA) in aqueous solution using a novel optical method. Biorecognition phage monolayers transferred to glass substrates using Langmuir-Blodgett (LB) technique were exposed individually to MRSA in solution at logarithmic concentrations ranging from 10(6) to 10(9)cfu/ml, and observed for real-time binding using a CytoViva optical light microscope system. Results indicate that LB monolayers possessed high levels of elasticity (K), measuring 22 and 29 mN/m for 10(9) and 10(11)pfu/ml phage concentrations, respectively. Near-instantaneous MRSA-phage binding produced 33+/-5%, 10+/-1%, 1.1+/-0.1%, and 0.09+/-0.01% coverage of the substrate that directly correlated to a decrease in MRSA concentrations of 10(9), 10(8), 10(7), and 10(6)cfu/ml. The exclusive selectivity of phage monolayers was verified with Salmonella enterica subsp. enterica serovar typhimurium (S. typhimurium) and Bacillus subtilis. PMID- 18424126 TI - A novel nitrite biosensor based on the direct electron transfer of hemoglobin immobilized on CdS hollow nanospheres. AB - A novel nitrite biosensor based on the direct electron transfer of hemoglobin (Hb) immobilized on CdS hollow nanospheres (HS-CdS) modified glassy carbon electrode was constructed. The direct electron transfer of Hb showed a pair of redox peaks with a formal potential of -286 mV (vs. SCE) in 0.1M pH 7.0 phosphate buffer solution. It was a surface-controlled electrode process involving a single proton transfer coupled with a reversible one-electron transfer for each heme group of Hb. HS-CdS had a large specific surface area and good biocompatibility and had a better electrochemical response than that of solid spherical CdS. The immobilized Hb on HS-CdS displayed an excellent response to NO(2)(-) with one irreversible electrode process for NO reduction. Under optimal conditions, the biosensor could be used for the determination of NO(2)(-) with a linear range from 0.3 to 182 microM and a detection limit of 0.08 microM at 3 sigma based on the irreversible reduction of NO. HS-CdS provided a good matrix for protein immobilization and had a promising application in constructing sensors. PMID- 18424127 TI - Towards beyond-1 GHz solution NMR: internal 2H lock operation in an external current mode. AB - We have commenced a project to develop a beyond-1 GHz solution NMR spectrometer using a HTS coil. Due to a small residual resistance present in the HTS conductor and joint resistance between conductors, a stable persistent current sufficient for NMR measurements is unlikely. Therefore, a current has to be supplied to the HTS coil from an external power supply. The ripple of an external power supply causes a field fluctuation which must be stabilized. In this study we show results of NMR measurements using a 500-600 MHz NMR in such an external current mode: the field fluctuations are stabilized by an internal 2H lock. The field fluctuation from the external power supply comprises a major field fluctuation component at low frequencies, 0.003-0.005 Hz, and superimposed minor field ripples at 2 Hz and 50 Hz. The former limits the time interval of the internal 2H lock, while the latter generates sidebands in the NMR spectrum. Sideband and baseline noise are controlled by appropriate selection of the feedback loop parameters of the lock. The quality of the 1D-solution NMR spectra observed in external current mode is equivalent to that obtained in persistent current mode. However, if the feedback loop time is as short as the gradient pulse width, refocusing of the NMR signal is lost and NMR peaks disappear. The 2D-NOESY and the 2D-HSQC spectra of ubiquitin in an external current mode have been acquired. The quality of the 2D spectra is equivalent to those obtained in persistent current mode; i.e. the internal 2H lock operates stably over an experimental time interval of 40-50 min. To realize a beyond-1 GHz NMR spectrometer, further investigations must be made of (i) the long term stability of a DC power supply, (ii) the enhancement of the compensation field limit for the internal 2H lock, (iii) the extension of the helium refill time interval, and (iv) a method to correct the field homogeneity in the external current mode. PMID- 18424128 TI - Probing the validity of average Hamiltonian theory for spin I=1, 3/2 and 5/2 nuclei by analyzing a simple two-pulse sequence. AB - In this work, we investigate the accuracy of controlling spin I=1, 3/2 and 5/2 spin systems by average Hamiltonian theory. By way of example, we consider a simple two-pulse echo sequence and compare this perturbation scheme to a numerical solution of the Von Neumann equation. For the different values of I, we examine this precision as a function of the quadrupolar coupling as well as various experimental parameters such as the pulse spacing and pulse width. Experiments and simulations on I=3/2 and I=5/2 spin systems are presented that highlight a spectral artifact introduced due to finite pulse widths as predicted by average Hamiltonian theory. The control of these spin systems by this perturbation scheme is considered by investigating a phase cycling scheme that suppresses these artifacts to zeroth-order of the Magnus expansion. PMID- 18424129 TI - The prognostic impact of functional imaging with (123)I-mIBG in patients with stage 4 neuroblastoma >1 year of age on a high-risk treatment protocol: results of the German Neuroblastoma Trial NB97. AB - AIM/PURPOSE: (123)I-meta-iodobenzylguanidine ((123)I-mIBG) scintigraphy is well established for staging and evaluation of response in children with high-risk neuroblastoma but its prognostic value in highly intensive first-line treatment protocols is uncertain. The presence of any (123)I-mIBG positive tumour tissue was correlated with event-free survival (EFS) and overall survival (OS). PATIENTS AND METHODS: The prognostic impact of residual (123)I-mIBG uptake into the primary tumour and metastases for predicting outcome in 113 stage 4 neuroblastoma patients >1 year of the German Neuroblastoma Trial NB97 was assessed using a univariate log-rank test and multivariate Cox regression analysis. RESULTS: All patients had (123)I-mIBG positive disease at initial staging. After four courses of induction chemotherapy, 71% of patients were still (123)I-mIBG positive for the primary tumour and 61% for metastases. After six courses, 39% of patients had (123)I-mIBG uptake by the primary tumour and 45% residual (123)I-mIBG positive metastatic disease. The (123)I-mIBG status of the primary tumour site had no bearing on outcome. Residual (123)I-mIBG positive metastatic disease after four (3-y-EFS 25.7+/-5.3% versus 55.9+/-7.6%, p=0.009; 3-y-OS 49.8+/-6.1% versus 65.0+/-7.3%; p=0.021) and after six chemotherapy cycles (3-y-EFS 27.5+/-6.2% versus 47.4+/-6.4%, p=0.011; 3-y-OS 50.5+/-7.1% vs 60.0+/-6.4%, p=0.031) was associated with poor outcome. CONCLUSION: Functional imaging with (123)I-mIBG scintigraphy can identify poor responders with any persistent metastatic (123)I mIBG uptake who are at a high risk of disease relapse. (123)I-mIBG response of the primary tumour site had no bearing on outcome. PMID- 18424130 TI - Production of nitric oxide by carp (Cyprinus carpio L.) kidney leukocytes is regulated by cyclic 3',5'-adenosine monophosphate. AB - The inducible nitric oxide synthase (iNOS) plays a central role in the inflammatory reactions that follow infection or tissue damage. Induction of nitric oxide (NO) synthesis by bacterial lipopolysaccharide (LPS) depends on activation of G protein-coupled receptors in mammals. Thus, it was our intention to evaluate whether similar mechanisms are involved in iNOS activation in fish leukocytes. Therefore, the participation of membrane-bound receptors which activate effectors via G proteins has been confirmed using the G protein inhibitor suramin. Furthermore, the NO produced by iNOS performs both beneficial and detrimental actions. It is thus conceivable that regulatory mechanisms exist which control the timing and intensity of NO production by iNOS in order to outweigh protective effects against detrimental ones. The second messenger cAMP produced by adenylyl cyclases (ACs) plays a key role in the regulation of many cellular functions. Since cAMP signaling inhibits numerous immunological reactions, studies have been carried out to determine whether cAMP-dependent pathways could inhibit NO production by carp leukocytes as well. To measure cellular responses such as NO production by carp leukocytes derived from head and trunk kidneys treatments were performed with the cAMP elevating agents forskolin and dibutyryl-cAMP (db-cAMP) prior to stimulation with Aeromonas hydrophila. Pharmacological studies in stimulated kidney leukocytes showed that increased intracellular cAMP levels lead to reduced NO formation. This reduction of NO production was not due to decreased cell numbers, since a tetrazolium dye-based assay revealed no reduction of cell viability by cyclic nucleotide elevating agents. Thus, our data provide evidence that the AC/cAMP signaling pathway is well established in carp leukocytes. Cyclic AMP leads to type II immune response. We provide evidence that the predominant AC in fish leukocytes is a particulate enzyme due to its sensitivity to forskolin. Treatment of leukocytes with agents increasing intracellular cAMP gave clear evidence for participation of this cyclic nucleotide in immune signaling. PMID- 18424131 TI - QSAR study of mosquito repellents from terpenoid with a six-member-ring. AB - A new class of low-toxicity mosquito repellents is synthesized from alpha- and beta-pinene in terpenoid compounds and preliminary biological tests show promising mosquito repellency. Statistical modeling is built using Codessa in order to reveal the quantitative relationship between the structure and the biological activity and to provide the guidance for further synthetic work. PMID- 18424132 TI - Novel echinocandin antifungals. Part 2: Optimization of the side chain of the natural product FR901379. Discovery of micafungin. AB - Further optimization of the potent antifungal activity of side chain analogs of the natural product FR901379 led to the discovery of compound 8 with an excellent, well-balanced profile. Potent compounds with reduced hemolytic potential were designed based upon a disruption of the linearity of the terphenyl lipophilic side chain. The optimized compound (8, FK463, micafungin) displayed the best balance and was selected as the clinical candidate. PMID- 18424133 TI - E-ring-modified 7-oxyiminomethyl camptothecins: Synthesis and preliminary in vitro and in vivo biological evaluation. AB - In contrast to five-membered E-ring analogues, 7-oxyiminomethyl derivatives of homocamptothecins showed ability to form stable ternary complexes with DNA and topoisomerase I. The 7-oxyiminomethyl derivatives of homocamptothecins were evaluated as a racemic mixture. Following the isolation of the two enantiomers, the 20 (R)-hydroxy isomer confirms the best activity. By using a panel of human tumor cells, all tested homocamptothecins showed a potent antiproliferative activity, correlating to the persistence of the cleavable complex. No significant difference was observed between the natural scaffold and the corresponding homocamptothecin homologue. A selected compound of this series exhibited an excellent antitumor activity against human gastrointestinal tumor xenografts. PMID- 18424134 TI - New tetrazole-based selective anandamide uptake inhibitors. AB - A new series of 1,5- and 2,5-disubstituted tetrazoles have been synthesized and evaluated as inhibitors of anandamide cellular uptake. Some of them inhibit the uptake process with a relatively high potency (IC(50)=2.3-5.1microM) and selectively over other proteins involved in endocannabinoid action and metabolism. PMID- 18424135 TI - Design of (N)-methanocarba adenosine 5'-uronamides as species-independent A3 receptor-selective agonists. AB - 2-Chloro-5'-N-methylcarboxamidoadenosine analogues containing the (N) methanocarba (bicyclo[3.1.0]hexane) ring system as a ribose substitute display increased selectivity as agonists of the human A(3) adenosine receptor (AR). However, the selectivity in mouse was greatly reduced due to an increased tolerance of this ring system at the mouse A(1)AR. Therefore, we varied substituents at the N(6) and C2 positions in search of compounds that have improved A(3)AR selectivity and are species independent. An N(6)-methyl analogue was balanced in affinity at mouse A(1)/A(3)ARs, with high selectivity in comparison to the A(2A)AR. Substitution of the 2-chloro atom with larger and more hydrophobic substituents, such as iodo and alkynyl groups, tended to increase the A(3)AR selectivity (up to 430-fold) in mouse and preserve it in human. Extended and chemically functionalized alkynyl chains attached at the C2 position of the purine moiety preserved A(3)AR selectivity more effectively than similar chains attached at the 3-position of the N(6)-benzyl group. PMID- 18424136 TI - Antidiabetic activity of N-(6-substituted-1,3-benzothiazol-2 yl)benzenesulfonamides. AB - N-(6-Substituted-1,3-benzothiazol-2-yl)benzenesulfonamide derivatives 1-8 were synthesized and evaluated for their in vivo antidiabetic activity in a non insulin-dependent diabetes mellitus rat model. Several compounds synthesized showed significant lowering of plasma glucose level in this model. As a possible mode of action, the compounds were in vitro evaluated as 11beta-hydroxysteroid dehydrogenase type 1 (11beta-HSD1) inhibitors. The most active compounds (3 and 4) were docked into the crystal structure of 11beta-HSD1. Docking results indicate potential hydrogen bond interactions with catalytic amino acid residues. PMID- 18424137 TI - Early and presymptomatic detection of Wilson's disease at the mandatory 3-year old medical health care examination in Hokkaido Prefecture with the use of a novel automated urinary ceruloplasmin assay. AB - Wilson's disease (WND) is an autosomal recessive disorder of copper (Cu) accumulation leading to liver and/or brain damage. Oral chelating agents and diet are effective in treating WND. However, once irreversible damage has occurred, the effect of treatment is diminished and the patient's quality of life is compromised. For these reasons an effective method for screening has been needed for early detection of presymptomatic patients. We conducted an early and presymptomatic detection of WND using a novel automated assay of ceruloplasmin (Cp) concentration in urine and selected the mandatory medical health care examination for 3-year-old children in Hokkaido Prefecture (the largest administrative division in Japan) as a sampling point. We measured urinary Cp concentrations in 11,362 children using an immunological latex agglutination assay kit developed by us. Among these children we identified a positive case with markedly reduced urinary Cp concentration. Detailed medical examination provided no clinical manifestations to support the diagnosis of WND, although serum Cp and Cu levels were remarkably low in this case. Therefore, we analyzed the WND gene in order to confirm the diagnosis. Sequence analysis revealed that the case was compound heterozygous for the WND gene mutations 2871del.C and D1296N. According to the Ferenci scoring system for WND diagnosis, the case was established as a WND patient at the presymptomatic stage. Consequently, the patient has maintained a good quality of life under medical treatment with polaprezinc administration to date. Our investigation suggests that the screening system for WND using the automated urinary assay at the mandatory medical health care examination for 3-year-old children is a noninvasive and efficient method for the early and presymptomatic diagnosis of WND. PMID- 18424138 TI - Treatment of Fabry disease with different dosing regimens of agalsidase: effects on antibody formation and GL-3. AB - Two different enzyme preparations are used for the treatment of Fabry disease patients, agalsidase alpha (Replagal, Shire) and agalsidase beta (Fabrazyme, Genzyme). Therapeutic efficacy of both products has been variable probably due to differences in gender, severity, age and other patient characteristics. We studied the occurrence of alpha-Gal A antibodies and their effect on urinary and plasma globotriaosylceramide (GL-3), plasma chitotriosidase and clinical outcome in 52 patients after 12 months of treatment with either 0.2mg/kg agalsidase alppha (10 males, 8 females) or beta (8 males, 5 females) or 1.0mg/kg agalsidase beta (10 males, 11 females). Antibodies were detected in 18/28 male patients after 6 months. None of the females developed antibodies. Following 12 months of 0.2mg/kg treatment, urinary GL-3 decreased in antibody negative (AB-) but increased in antibody positive (AB+) patients. Treatment with 1.0mg/kg gave a reduction in urinary GL-3 in both AB- and AB+ patients. Levels of plasma GL-3 and chitotriosidase decreased in all patient groups. Twelve months of 0.2mg/kg treatment did not change renal function or left ventricular mass. Further, no change in renal function was seen following 1.0mg/kg treatment and left ventricular mass decreased in both AB- and AB+ patients. In summary, alpha-Gal A antibodies frequently develop in male Fabry disease patients and interfere with urinary GL-3 excretion. Infusion of a dose of 1.0mg/kg results in a more robust decline in GL-3, less impact, if any of antibodies, stable renal function and reduction of LVMass. PMID- 18424139 TI - A lungless frog discovered on Borneo. PMID- 18424140 TI - Natural variation in leaf morphology results from mutation of a novel KNOX gene. AB - Striking diversity in size, arrangement, and complexity of leaves can sometimes be seen in closely related species. One such variation is found between wild tomato species collected by Charles Darwin from the Galapagos Islands [1-5]. Here, we show that a single-nucleotide deletion in the promoter of the PETROSELINUM (PTS) [3] gene upregulates the gene product in leaves and is responsible for the natural variation in leaf shape in the Galapagean tomatoes. PTS encodes a novel KNOTTED1-LIKE HOMEOBOX (KNOX) gene that lacks a homeodomain. We also showed that the tomato classical mutant bipinnata (bip) [6], which recapitulates the Pts phenotype, results from the loss of function of a BEL-LIKE HOMEODOMAIN (BELL) gene, BIP. We used bimolecular fluorescence complementation and two-hybrid competition assays to show that PTS represses KNOX1 protein interactions with BIP, as well as subsequent nuclear localization of this transcriptional complex. We suggest that natural variation in leaf shape can be created with a rheostat-like mechanism that alters the KNOX1 protein interaction network specifically during leaf development. This subtle change in interaction between transcription factors leaves essential KNOX1 function in the shoot apical meristem intact and appears to be a facile way to alter leaf morphology during evolution. PMID- 18424141 TI - Polarized subcellular localization of Jak/STAT components is required for efficient signaling. AB - Three protein complexes control polarization of epithelial cells: the apicolateral Crumbs and Par-3 complexes and the basolateral Lethal giant larvae complex. Polarization results in the specific localization of proteins and lipids to different membrane domains. The receptors of the Notch, Hedgehog, and WNT pathways are among the proteins that are polarized, with subcellular receptor localization representing an important aspect of signaling regulation. For example, in the WNT pathway, differential DFz2 receptor localization results in activation of either the canonical or the planar polarity pathway. Despite the large body of research on the vertebrate JAK/STAT pathway, there are no reports indicating polarized signaling. By using the conserved Drosophila JAK/STAT pathway as a system, we find that the receptor and its associated kinase are located in the apical membrane of epithelial cells. Unexpectedly, the transcription factor STAT is enriched in the apicolateral membrane domain of ectoderm epithelial cells in a Par-3-dependent manner. Our results indicate that preassembly of STAT and the Receptor/JAK complex to specific membrane domains is a key aspect for signaling efficiency. Our results also suggest that receptor polarization in the ectoderm cell membrane restricts the cell's response to ligands provided by neighboring cells. PMID- 18424142 TI - The direct assessment of genetic heterozygosity through scent in the mouse. AB - The role of individual genetic heterozygosity in mate choice is the subject of much current debate. Several recent studies have reported female preference for more heterozygous males, but the mechanisms underlying heterozygote preference remain largely unknown. Females could favor males that are more successful in intrasexual competition, but they could also assess male heterozygosity directly at specific polymorphic genetic markers. Here, we use a breeding program to remove the intrinsic correlation between genome-wide heterozygosity and two highly polymorphic gene clusters that could allow direct assessment of heterozygosity through scent in mice: the major histocompatibility complex (MHC) and the major urinary proteins (MUPs). When other sources of variation are controlled and intrasexual competition is minimized, female mice prefer to associate with MUP heterozygous over MUP homozygous males. MHC heterozygosity does not influence preference, and neither does heterozygosity across the rest of the genome when intrasexual competition between males is restricted. Female mice thus assess male heterozygosity directly through multiple MUP isoforms expressed in scent signals, independently of the effects of genome-wide heterozygosity on male competitiveness. This is the first evidence that animals may use signals of genetic heterozygosity that have no direct association with individual vigour. PMID- 18424143 TI - Directional prediction by the saccadic system. AB - One popular and fruitful approach to understanding what influences the decision of where to look next has been to present targets in a series of trials either to the right or left of a central fixation point and examine sequential effects on saccadic latency. However, there is a problem with this paradigm: Every saccade to a target is necessarily followed by an equal and opposite movement back to the center, yet the potentially confounding influence of this refixation saccade is rarely considered. Here, we introduce a novel random-walk paradigm that eliminates this difficulty. Each successive target appears to the left or right of the previous one, allowing us to study long sequences of saccades uncontaminated by refixations. This exposes a new stimulus-history effect, which is remarkably prolonged and relates primarily to movement direction: A saccade reduces the latency for subsequent movements made in the same direction and retards those in the opposite direction. Although in conventional refixation paradigms this effect cancels out, it is of particular significance in the real world--where our fixation point shifts constantly with the object of interest- and reflects a prediction of the way that real objects typically move. PMID- 18424144 TI - Orientation of birds in total darkness. AB - Magnetic compass orientation of migratory birds is known to be light dependent, and radical-pair processes have been identified as the underlying mechanism. Here we report for the first time results of tests with European robins, Erithacus rubecula, in total darkness and, as a control, under 565 nm green light. Under green light, the robins oriented in their normal migratory direction, with southerly headings in autumn and northerly headings in spring. By contrast, in darkness they significantly preferred westerly directions in spring as well as autumn. This failure to show the normal seasonal change characterizes the orientation in total darkness as a "fixed direction" response. Tests in magnetic fields with the vertical or the horizontal component inverted showed that the preferred direction depended on the magnetic field but did not involve the avian inclination compass. A high-frequency field of 1.315 MHz did not affect the behavior, whereas local anesthesia of the upper beak resulted in disorientation. The behavior in darkness is thus fundamentally different from normal compass orientation and relies on another source of magnetic information: It does not involve the radical-pair mechanism but rather originates in the iron-containing receptors in the upper beak. PMID- 18424145 TI - Fragment-based learning of visual object categories. AB - When we perceive a visual object, we implicitly or explicitly associate it with a category we know. It is known that the visual system can use local, informative image fragments of a given object, rather than the whole object, to classify it into a familiar category. How we acquire informative fragments has remained unclear. Here, we show that human observers acquire informative fragments during the initial learning of categories. We created new, but naturalistic, classes of visual objects by using a novel "virtual phylogenesis" (VP) algorithm that simulates key aspects of how biological categories evolve. Subjects were trained to distinguish two of these classes by using whole exemplar objects, not fragments. We hypothesized that if the visual system learns informative object fragments during category learning, then subjects must be able to perform the newly learned categorization by using only the fragments as opposed to whole objects. We found that subjects were able to successfully perform the classification task by using each of the informative fragments by itself, but not by using any of the comparable, but uninformative, fragments. Our results not only reveal that novel categories can be learned by discovering informative fragments but also introduce and illustrate the use of VP as a versatile tool for category-learning research. PMID- 18424146 TI - Subspecies genetic assignments of worldwide captive tigers increase conservation value of captive populations. AB - Tigers (Panthera tigris) are disappearing rapidly from the wild, from over 100,000 in the 1900s to as few as 3000. Javan (P.t. sondaica), Bali (P.t. balica), and Caspian (P.t. virgata) subspecies are extinct, whereas the South China tiger (P.t. amoyensis) persists only in zoos. By contrast, captive tigers are flourishing, with 15,000-20,000 individuals worldwide, outnumbering their wild relatives five to seven times. We assessed subspecies genetic ancestry of 105 captive tigers from 14 countries and regions by using Bayesian analysis and diagnostic genetic markers defined by a prior analysis of 134 voucher tigers of significant genetic distinctiveness. We assigned 49 tigers to one of five subspecies (Bengal P.t. tigris, Sumatran P.t. sumatrae, Indochinese P.t. corbetti, Amur P.t. altaica, and Malayan P.t. jacksoni tigers) and determined 52 had admixed subspecies origins. The tested captive tigers retain appreciable genomic diversity unobserved in their wild counterparts, perhaps a consequence of large population size, century-long introduction of new founders, and managed breeding strategies to retain genetic variability. Assessment of verified subspecies ancestry offers a powerful tool that, if applied to tigers of uncertain background, may considerably increase the number of purebred tigers suitable for conservation management. PMID- 18424147 TI - Cooperative sentinel calling? Foragers gain increased biomass intake. AB - Many foraging animals face a fundamental tradeoff between predation and starvation. In a range of social species, this tradeoff has probably driven the evolution of sentinel behavior, where individuals adopt prominent positions to watch for predators while groupmates forage. Although there has been much debate about whether acting as a sentinel is a selfish or cooperative behavior, far less attention has focused on why sentinels often produce quiet vocalizations (hereafter known as "sentinel calls") to announce their presence. We use observational and experimental data to provide the first evidence that group members gain an increase in foraging success by responding to these vocal cues given by sentinels. Foraging pied babblers (Turdoides bicolor) spread out more, use more exposed patches, look up less often, and spend less time vigilant in response to sentinel calling. Crucially, we demonstrate that these behavioral alterations lead to an increase in biomass intake by foragers, which is likely to enhance survival. We argue that this benefit may be the reason for sentinel calling, making it a truly cooperative behavior. PMID- 18424148 TI - Autologous blood predonation in cardiac surgery. AB - The risk of transfusion of allogeneic blood products on outcome is well documented. Autologous blood donation prior to elective cardiac surgery has repeatedly shown to be an effective practice to reduce the exposure to allogeneic blood, but was criticized because of the alleged high costs. We analyzed the data of 4878 patients undergoing elective open-heart surgery, in whom 18% underwent autologous donation. Overall, autologous blood donation reduced the incidence of allogeneic blood transfusion from 48 to 13% during hospitalization. Additionally, it is shown that diagnosis adjusted autologous blood donation is cost-effective. The higher the probability of transfusion the better is efficacy and cost effectiveness of predonation. Cardiac surgery is a high-transfusion area, thus, it offers ideal conditions for autologous donation. Autologous blood donation still remains a promising and cost-effective alternative to reduce allogeneic blood transfusion in elective cardiac surgery. PMID- 18424149 TI - Posturographic assessment of sitting balance recovery in the subacute phase of stroke. AB - Although early sitting balance is a well-known predictor of functional outcome after stroke, it is still unknown which aspects of normal upright sitting balance are most sensitive to subsequent recovery. This study used an adjustable chair mounted on a force platform to assess the recovery of quiet-sitting balance in 16 patients with a first supratentorial stroke during their inpatient rehabilitation. The patients underwent three posturographic assessments at 6-week intervals from the moment of their admission, on average 5.6 weeks after stroke. Each quiet-sitting balance assessment consisted of two series of four 30-s test conditions: sitting with eyes open and closed, on both a stable and unstable (air cushion) surface. The RMS of the center-of-pressure (COP) velocities was used as the primary measure of lateral and anteroposterior balance control. It was found that, compared to 10 healthy elderly, lateral balance was more affected by stroke than balance in the anteroposterior direction, especially during visual deprivation, and most sensitive to subsequent functional changes induced by spontaneous recovery or rehabilitation. Furthermore, lateral balance control showed the strongest association with the Berg Balance Scale as a clinical measure of balance capacity. Hence, (lateral) trunk control seems to be a primary target for rehabilitation. Since an unstable support was necessary to obtain significant effects of stroke, recovery and visual deprivation, it may be important to use an unstable support during sitting balance training as well. PMID- 18424150 TI - [Endoscopic anatomy of the terminal portion of the basilar artery and its distal perforating branches]. AB - The aim of this study was to describe and highlight the endoscopic anatomy of the tip of the basilar artery and its perforating branches. Knowledges of the anatomy are crucial for neurosurgeons to avoid pitfalls during endoscopic third ventriculostomy. PMID- 18424151 TI - Concurrent presentation of ocular myasthenia and euthyroid Graves ophthalmopathy: a diagnostic challenge. PMID- 18424152 TI - New method of bone reconstruction designed for skull base surgery. AB - The direct endonasal or transoral transclival approaches to the skull base permit effective, minimally invasive surgery along the clivus. Developing long-term, effective techniques to prevent cerebrospinal fluid (CSF) leaks and their consequences (infection and delayed healing) remains a major challenge. In this study we describe a method of bone reconstruction newly developed by us, which uses a custom designed silicone plug for bone replacement after minimally invasive skull base surgery with a low incidence of postoperative CSF leaks. German Landrace pigs were used to test the efficiency of the new technique. Twelve craniotomies were performed in six pigs using a subtemporal approach and subsequently the dura was opened. After these preparations the craniotomy defects were occluded with a silicone ball, which had a near spherical shape. The ball elastically adapts to the bone defect. Each pig also received an intracranial pressure (ICP) catheter and a subdural catheter for later fluorescein injection. Then we increased ICP by infusion of artificial CSF and detected fluorescein leaks from the craniotomy using ultraviolet illumination and a photomacroscope equipped with appropriate filters and a charge-coupled device camera. In all pigs we increased ICP to 75-80 mmHg by infusing 25-30 mL saline containing 0.05% sodium fluorescein. For the first four craniotomies infusions were interrupted after CSF leaks occurred due to technical failures, which were subsequently rectified. The following eight craniotomies were watertight without CSF leakage. This novel medical device allows a leak-proof closure of bone defects after minimally invasive craniotomies; no additional surgery or other therapies were necessary. The application of the silicone plug, which is made of a cost effective and biocompatible material, is easy and fast, making use of a specially developed toolkit. PMID- 18424153 TI - Novel human 3-domain disulfide-stabilized antibody fragment against glycoprotein of rabies virus. AB - Mutated disulfide bond sites VH (Cys44) and VL (Cys100) were constructed in variable domains (Fvs) of the human anti-glycoprotein antigen of the rabies virus (anti-GPRV), and the light chain variable (VL) and heavy chain variable (VH) fragments were linked using the heavy chain constant region 1 (CH1) of the human immunoglobulin (Ig) to successfully construct a 3-domain disulfide-stabilized fragment of variables (3d-dsFv). 3d-dsFv was mainly expressed as an inclusion body. After refolding by the conventional dilution method, 3d-dsFv was purified using a nickel-nitrilotriacetic acid (Ni-NTA) column. Enyzme-linked immunosorbent assay (ELISA) was used to determine the binding activity of 3d-dsFv to GPRV. Flow cytometry studies and rapid fluorescent focus inhibition test were used to evaluate the function of 3d-dsFv. The results showed that the stability of 3d dsFv was improved notably in some aspects such as thermal kinetics, ability to withstand urea denaturation, etc. 3d-dsFv could bind specially to infective cells and the GPRV. The titration of 3d-dsFv to RV-CVS is 83.3 IU/mg, and it can easily reach 2.5IU/mL, which is the value suggested by the WHO as effective for neutralization titration of the rabies virus. PMID- 18424154 TI - Waddlia chondrophila enters and multiplies within human macrophages. AB - Waddlia chondrophila is an obligate intracellular bacterium of the Chlamydiales order. W. chondrophila has been isolated twice from aborted bovine foetuses and a serological study supported the abortigenic role of W. chondrophila in bovine species. Recently, we observed a strong association between the presence of anti Waddlia antibodies and human miscarriage. To further investigate the pathogenic potential of W. chondrophila in humans, we studied the entry and the multiplication of this Chlamydia-like organism in human macrophages. Confocal and electron microscopy confirmed that W. chondrophila is able to enter human monocyte-derived macrophages. Moreover, W. chondrophila multiplied readily within macrophages. The proportion of infected macrophages increased from 13% at day 0 to 96% at day 4, and the mean number of bacteria per macrophage increased by 3logs in 24h. Intracellular growth of W. chondrophila was associated with a significant cytopathic effect. Thus, W. chondrophila may enter and grow rapidly within human macrophages, inducing lysis of infected cells. Since macrophages are one of the major components of the innate immune response, these findings indirectly suggest the possible human pathogenicity of W. chondrophila. PMID- 18424155 TI - Synthesis, cytostatic and anti-HIV evaluations of the new unsaturated acyclic C-5 pyrimidine nucleoside analogues. AB - A series of the novel C-5 alkynyl pyrimidine nucleoside analogues (1-14) in which the sugar moiety was replaced by the conformationally restricted Z- and E-2 butenyl spacer between the phthalimido and pyrimidine ring were synthesized by using Sonogashira cross-coupling reaction. Cytostatic activity evaluation of the novel compounds showed that E-isomers exhibited, in general, better cytostatic activities than the corresponding Z-isomers. E-isomer 14 exhibited the best cytostatic effect against all evaluated malignant cell lines, particularly against hepatocellular carcinoma (Hep G2, IC(50)=4.3microM). However, this compound was also cytotoxic to human normal fibroblasts (WI 38). Its Z-isomer 7 showed highly specific antiproliferative activity against Hep G2 (IC(50)=18microM) and no cytotoxicity to WI 38. Moreover, compounds 3, 4 and 14 expressed some marginal inhibitory activity against HIV-1 and HIV-2. PMID- 18424156 TI - Synthesis and cytotoxic properties of new fluorodeoxyglucose-coupled chlorambucil derivatives. AB - Frequently used in the treatment of malignant cells, alkylating agents, like most anticancer substances, produce adverse side effects caused by the toxicity of the agents toward normal tissues and lose efficiency through poor distribution to target sites. Our approach to developing more selective drugs with low systemic toxicity is based on the premise that the body distribution and cell uptake of a drug can be altered by attaching a neoplastic cell-specific uptake enhancer, such as 2-fluoro-2-deoxyglucose (FDG), the radiotracer most frequently used in PET for tumor imaging. Two properties of deoxyglucose, namely preferential accumulation in neoplastic cells and inhibition of glycolysis, underpin this targeting approach. Here, we report the synthesis of 19 new chlorambucil glycoconjugates in which the alkylating drug is attached to the C-1 position of FDG, directly or via different linkages. This set of compounds was evaluated for in vitro cytotoxicity against different human normal and tumor cell lines. There was a significant improvement in the in vitro cytotoxicity of peracetylated glucoconjugates compared with the free substance. Four compounds were finally selected for further in vivo studies owing to their lack of oxidative stress-inducing properties. PMID- 18424157 TI - Synthetic methods for the preparation of ARQ 501 (beta-Lapachone) human blood metabolites. AB - ARQ 501 (3,4-dihydro-2,2-dimethyl-2H-naphthol[1,2-b] pyran-5,6-dione), a synthetic version of beta-Lapachone, is a promising anti-cancer agent currently in multiple Phase II clinical trials. Promising anti-cancer activity was observed in Phase I and Phase II trials. Metabolism by red blood cells of drugs is an understudied area of research and the metabolites arising from oxidative ring opening (M2 and M3), decarbonylation/ring contraction (M5), and decarbonylation/oxidation (M4 and M6) of ARQ 501 offer a unique opportunity to provide insight into these metabolic processes. Since these metabolites were not detected in in vitro incubations of ARQ 501 with liver microsomes and were structurally diverse, confirmation by chemical synthesis was considered essential. In this report, we disclose the synthetic routes employed and the characterization of the reference standards for these blood metabolites as well as additional postulated structures, which were not confirmed as metabolites. PMID- 18424158 TI - A practice survey of shoulder glucocorticoid injections in patients on antiplatelet drugs or vitamin K antagonists. AB - OBJECTIVE: Little is known about practice patterns regarding shoulder glucocorticoid injections in patients taking antiplatelet drugs or vitamin K antagonists. The objective of this study was to collect relevant data among rheumatologists in France. METHODS: We conducted a postal questionnaire survey among 2015 rheumatologists. RESULTS: We received 1018 completed questionnaires (response rate, 1018/2015, 50.5%). The proportion of rheumatologists who discontinued drugs with antithrombotic effects prior to shoulder injections varied across drugs, as follows: nonsteroidal antiinflammatory drugs (NSAIDs), 2.6%; dipyridamole, 7.5%; aspirin, 9%; ticlopidine, 25%; clopidogrel, 28%; and vitamin K antagonists, 74%. Among rheumatologists who discontinued vitamin K antagonist therapy, 82% prescribed replacement therapy with low-molecular-weight heparin. Time from discontinuation to injection was not consistently appropriate to the duration of drug effects. Prior to the injection, hemostasis tests were obtained by 1% of rheumatologists for patients on NSAIDs, 6% for those on clopidogrel, and 65% for those on vitamin K antagonists. Only 1% of rheumatologists had observed bleeding events in patients on aspirin, compared to 10% in patients on vitamin K antagonists. The mean number of shoulder glucocorticoid injections per rheumatologist per month was 19.6; 4% of rheumatologists routinely obtained hemostasis tests before shoulder injections. CONCLUSION: Considerable variations were found among rheumatologists regarding practice patterns for performing shoulder glucocorticoid injections. This variability reflects the absence of official guidelines. The GREP is working with French hematologists to develop clinical practice guidelines. PMID- 18424159 TI - Low frequency of HLA-B27 in ankylosing spondylitis patients from Turkey. AB - OBJECTIVES: Ankylosing spondylitis is strongly associated with HLA-B27. However, the strength of the association with HLA-B27 and the clinical features may vary in different parts of the world. The aim of this study is to compare the clinical features of AS and the frequencies of HLA-B27 and its alleles in patients from Turkey with other series. METHODS: One hundred and twelve patients (72 male/40 female) fulfilling the modified New York criteria for the classification of AS and 55 (27 male/28 female) healthy controls were typed for HLA-B27 and allele frequencies by sequence specific primer (PCR/SSP) method and assessed for clinical manifestations. RESULTS: Male to female ratio was 1.8, mean age at disease onset was 23.5 and 24.1% of patients reported juvenile onset of symptoms. Peripheral arthritis was seen in 52.7% of patients. Family history (p=0.01) and peripheral arthritis (p=0.02) were more frequent in females and spinal involvement in males. HLA-B27 was found to be positive in 70% of patients and associated with younger mean age, uveitis and shorter time elapsed from symptom to diagnosis. The frequency of HLA-B27 alleles associated with SpA was not different between ankylosing spondylitis patients and healthy controls. CONCLUSION: Low frequency of HLA-B27 and clinical variations in ankylosing spondylitis may be due to different genetic and/or environmental factors in Turkey. PMID- 18424160 TI - Organizing pneumonia associated with rituximab: Challenges raised by establishing causality. PMID- 18424161 TI - Beneficial effects of rolipram in the R6/2 mouse model of Huntington's disease. AB - We have previously showed that rolipram, a phosphodiesterase type IV inhibitor, displays a neuroprotective effect in a rat quinolinic acid model of HD [DeMarch Z., Giampa C., Patassini S., Martorana A., Bernardi G. and Fusco F.R., (2007) Beneficial effects of rolipram in a quinolinic acid model of striatal excitotoxicity. Neurobiol. Dis. 25:266-273.]. In this study, we sought to determine if rolipram exerts a neuroprotective effect in R6/2 mutant mice, which recapitulates, in many aspects, human HD [Mangiarini L., Sathasivam K., Seller M., Cozens B., Harper A., Hetherington C., Lawton M., Trottier Y., Lehrach H., Davies S.W. and Bates G.P. (1996) Exon 1 of the HD gene with an expanded CAG repeat is sufficient to cause a progressive neurological phenotype in transgenic mice. Cell. 87:493-506]. Transgenic mice were treated with rolipram 1.5 mg/kg daily starting from 4 weeks of age. After transcardial perfusion, histological and immunohistochemical studies were performed. We found that rolipram-treated R6/2 mice survived longer and displayed less severe signs of neurological dysfunction than the vehicle treated ones. Primary outcome measures such as brain volume, striatal atrophy, size and morphology of striatal neurons, neuronal intranuclear inclusions and microglial reaction confirmed a neuroprotective effect of the compound. Rolipram was effective in increasing significantly the levels of activated CREB and of BDNF the striatal spiny neurons, which might account for the beneficial effects observed in this model. Our findings show that rolipram could be considered as a valid therapeutic approach for HD. PMID- 18424162 TI - [Role of ultrasound in the evaluation of patients after laparoscopic sacropexy: preliminary study]. AB - OBJECTIVES: To evaluate results of laparoscopic sacropexy with introital and vaginal ultrasonography, that is to observe dynamic behavior of the mesh, to measure precisely its position and to conclude on the feasibility of this exam. PATIENTS AND METHODS: The surgery was a laparoscopic subtotal hysterectomy and a suspension of the cervix with an anterior vesicovaginal mesh and a posterior rectovaginal mesh. These meshes are joined together and fixed on the sacrum. Ultrasound was performed in fifteen patients to see in the sagital plane the position of the mesh. The distance anterior mesh-bladder neck and the distance posterior mesh-anal sphincter were measured at rest, during straining and during retaining. The spreading out and the dynamic behavior of the mesh were studied. RESULTS: The distance anterior mesh-bladder neck is 12 mm at rest, 15 mm during straining and 13 mm during retaining. The distance posterior mesh-anal sphincter is 30 mm at rest, 26 mm during straining and 31 mm during retaining. The mesh is always seen as spread out, or folded, but never strained. Video recording shows that sacropexy does not block pelvic dynamics. DISCUSSION AND CONCLUSION: Ultrasound imaging appears in this study as a simple and efficient exam to study and measure prosthesis. It could be a good help for clinical examination to explain failure or complication of surgery. PMID- 18424163 TI - [Hormonal contraception and risk of venous thromboembolism: When to ask for an assessment of hemostasis? Which parameters?]. AB - One of the deleterious effects of the combined oral contraceptives is venous thromboembolism (VTE) and it is the most frequent. VTE is potentially serious because it is sometimes responsible for fatal pulmonary embolism. Because of the large use of hormonal contraception among healthy women and often for long durations, it is fundamental to target the prescriptions and detect women at high risk of VTE. It has been demonstrated that some congenital or acquired coagulation anomalies are associated with an increase of thromboembolic risk. In addition, combined oral contraceptives modify some parameters of the hemostasis, whatever the route of administration. In order to optimize the benefit-risk balance of oral contraception, the search for a biological thrombophilia is essential in some clinical situations such as in young women with a history of venous thromboembolic event or with a family history of thrombosis at a relatively young age. A thorough questioning must be performed. On the other hand, this biological research is not systematically recommended before any prescription of hormonal contraception in patients having neither previous personal nor family history of venous thrombosis. PMID- 18424165 TI - Development of human lymph nodes and Peyer's patches. AB - In contrast to our understanding of murine lymphoid organogenesis, detailed knowledge on the mechanisms of human lymph node development is virtually lacking. This is mainly due to the obvious difficulties that accompany research using human fetal organs. In this review we will highlight current knowledge on human lymph node and Peyer's patch development and will temporally align observations made in humans with data available from murine studies. In the final paragraphs we will put this knowledge in the context of human malignancies in which interactions between lymphocytes and stroma, resembling those seen in lymphoid organs, are recapitulated. PMID- 18424164 TI - Translocation of arrestin induced by human A(3) adenosine receptor ligands in an engineered cell line: comparison with G protein-dependent pathways. AB - Structurally diverse ligands were studied in A(3) adenosine receptor (AR) mediated beta-arrestin translocation in engineered CHO cells. The agonist potency and efficacy were similar, although not identical, to their G protein signaling. However, differences have also been found. MRS542, MRS1760, and other adenosine derivatives, A(3)AR antagonists in cyclic AMP assays, were partial agonists in beta-arrestin translocation, indicating possible biased agonism. The xanthine 7 riboside DBXRM, a full agonist, was only partially efficacious in beta-arrestin translocation. DBXRM was shown to induce a lesser extent of desensitization compared with IB-MECA. In kinetic studies, MRS3558, a potent and selective A(3)AR agonist, induced beta-arrestin translocation significantly faster than IB-MECA and Cl-IB-MECA. Non-nucleoside antagonists showed similar inhibitory potencies as previously reported. PTX pretreatment completely abolished ERK1/2 activation, but not arrestin translocation. Thus, lead candidates for biased agonists at the A(3)AR have been identified with this arrestin-translocation assay, which promises to be an effective tool for ligand screening. PMID- 18424166 TI - Development of a new design of hip protectors using finite element analysis and mechanical tests. AB - The rate of hip fractures due to falls increases with age. External hip protectors placed over the greater trochanter can prevent hip fractures, but the willingness to wear such protective devices is rather low. Most of the commercially available hip protectors consist either of an energy-absorbing foam pad or of a hard shell that distributes the load to the surrounding tissue. In the present study, a fibre-reinforced shell composed of three curved strips bonded with a ring, was designed and lined with shock absorbing foam. The development of the new shell design was based on quasi-static and impact tests of manufactured shells in combination with finite element simulations. The results of the numerical analysis showed the potential protection effect of the shell and indicated how the design can be further improved. First impact tests on an anatomical hip model showed promising results of the new protector shell in combination with a foam pad. PMID- 18424168 TI - Synaptopodin maintains the neural activity-dependent enlargement of dendritic spines in hippocampal neurons. AB - Synaptopodin (SYNPO) is an F-actin interacting protein expressed in dendritic spines and upregulated during the late-phase of long-term potentiation. Here, we investigated whether SYNPO regulates spine morphology through interactions with F actin, the major cytoskeletal element of spines. In primary hippocampal neuron cultures, both endogenous and exogenous SYNPO localized preferentially in large spines under basal conditions. SYNPO overexpression did not affect the number or volume of spines in unstimulated neurons. Pharmacological activation of synaptic NMDA receptors transiently increased spine volume in control neurons, while the increase was persistent in neurons overexpressing SYNPO. In addition, exogenous SYNPO in PtK2 cells suppressed staurosporine-dependent disruption of F-actin stress fibers, suggesting that SYNPO protected F-actin from disruption. These results suggest that SYNPO stabilized activity-dependent increases in spine volume and imply that late-phase changes in spine morphology involve SYNPO. PMID- 18424167 TI - Calcium-modulating cyclophilin ligand regulates membrane trafficking of postsynaptic GABA(A) receptors. AB - Accumulation of GABA(A) receptors (GABA(A)Rs) at GABAergic synapses requires the cytoplasmic loop region and C-terminal transmembrane domain of the receptor gamma2 subunit. We here report a novel interaction of gamma2 with Calcium Modulating cyclophilin Ligand (CAML), an integral membrane protein that regulates this mechanism. Interaction of GABA(A)Rs with CAML depends on both the cytoplasmic region and fourth transmembrane domain of the gamma2 subunit, CAML immunoprecipitates with GABA(A)Rs from transfected cells and brain lysates and colocalizes with gamma2 in ER vesicles in soma and dendrites of neurons. CAML shRNA treatment results in reduced expression of postsynaptic GABA(A)Rs, along with significant reductions in GABA-evoked whole cell currents and GABAergic synaptic function, while glutamatergic transmission is unaffected. Reduced surface expression of GABA(A)Rs in CAML mutant neurons is associated with selective deficits in recycling of endocytosed GABA(A)Rs to the cell surface. Our results indicate a specific role of CAML in functional expression and endocytic recycling of postsynaptic GABA(A)Rs. PMID- 18424169 TI - Dietary fats, cholesterol and iron as risk factors for Parkinson's disease. AB - BACKGROUND: Epidemiologic findings suggest that dietary components may contribute to the etiology of Parkinson's disease (PD). This population-based case-control study evaluated PD risk and dietary intake of fats, cholesterol and iron. METHODS: Newly diagnosed case (n=420) and age/gender/ethnicity-matched unrelated controls (n=560) were identified between 1992 and 2006 from the Group Health Cooperative health maintenance organization in western Washington State, and the University of Washington Neurology Clinic. In-person interviews elicited data on food frequency habits during most of adult life. Nutritional intakes were calculated and analyzed, with adjustments made for total energy intake (the 'nutrition density' technique). RESULTS: Cholesterol intake in the highest quartile compared with the lowest quartile was associated with a decreased risk of PD in men (odds ratio (OR)=0.53, 95%CI: 0.33, 0.86). The highest versus the lowest quartile of dietary iron increased PD risk in men (OR=1.82, 95%CI: 1.11, 2.99). When the lowest quartile of cholesterol and the highest quartile for iron were compared to the highest quartile of cholesterol and the lowest quartile of iron, no association was seen in women, but for men PD risk was increased (OR=2.70, 95%CI: 1.26, 5.76). Saturated fat intake below the median in combination with iron intake above the median also increased the PD risk (OR=1.50, 95%CI: 1.07, 2.11) in both genders combined. CONCLUSIONS: A low intake of cholesterol, particularly in the presence of high iron, may be associated with an increased risk for PD. PMID- 18424170 TI - Characterization of detergent purified recombinant rat liver monoamine oxidase B expressed in Pichia pastoris. AB - The high level expression and purification of rat monoamine oxidase B (rMAOB) in the methylotrophic yeast Pichia pastoris is reported. Nearly 100 mg of purified rMAOB is obtained from 130 g (wet weight) of cells (0.5 L of culture). The MALDI TOF mass spectrum of the purified protein shows a single species with a molecular mass of 59.228 +/- 0.064 kDa, which agrees with the calculated molecular weight of 59.172 kDa for the rMAOB protein sequence assuming one mole of covalent FAD per mole of the enzyme. Consistent with the MALDI-MS data, purified rMAOB shows a single band near 60 kDa in Coomassie-stained SDS-PAGE gel as well as on Western blot analyses performed using antisera raised against human MAOA and BSA conjugated FAD. A partial amino acid sequence of the purified protein is confirmed to be that of the wild type rMAOB by in-gel trypsin digestion and MALDI TOF-MS analyses of the liberated peptide fragments. Steady state kinetic data show that purified rMAOB exhibits a K(m)(amine) of 176 +/- 15 microM and a k(cat) of 497 +/- 83 min(-1) for benzylamine oxidation, and a K(m)(O2) of 170 +/- 10 microM. Kinetic parameters obtained for purified rMAOB are compared with those reported earlier for recombinant human liver MAOB expressed in P. pastoris. PMID- 18424171 TI - Revealing the structural and functional diversity of plant cell walls. AB - The extensive knowledge of the chemistry of isolated cell wall polymers, and that relating to the identification and partial annotation of gene families involved in their synthesis and modification, is not yet matched by a sophisticated understanding of the occurrence of the polymers within cell walls of the diverse cell types within a growing organ. Currently, the main sets of tools that are used to determine cell-type-specific configurations of cell wall polymers and aspects of cell wall microstructures are antibodies, carbohydrate-binding modules (CBMs) and microspectroscopies. As these tools are applied we see that cell wall polymers are extensively developmentally regulated and that there is a range of structurally distinct primary and secondary cell walls within organs and across species. The challenge now is to document cell wall structures in relation to diverse cell biological events and to integrate this knowledge with the emerging understanding of polymer functions. PMID- 18424172 TI - Inflammatory response to Dentitruncus truttae (Acanthocephala) in the intestine of brown trout. AB - Brown trout, Salmo trutta L., were infected with the acanthocephalan Dentitruncus truttae with the most affected areas being the anterior (near the pyloric caeca) and middle intestine. The parasite attached with a proboscis which usually penetrated the mucosa, lamina propria, stratum compactum, stratum granulosum and, sometimes, the muscularis layer. Around the parasite's body was an area of inflammatory tissue. At the point of attachment the lamina propria was thickened and the stratum compactum, stratum granulosum and muscularis layer were disrupted by proboscis penetration. Rodlet cells were more numerous in infected fish (P<0.01), and were found in the epithelial layer away from the worm. Infected intestines had larger numbers of mast cells (P<0.01), often in close proximity to, and inside, the blood capillaries and associated with fibroblasts of the muscularis layer and the stratum granulosum. Their migration toward the site of infection was suggested. Intense degranulation of mast cells was encountered in all intestinal layers especially near the parasite's body. Immunohistochemical tests were conducted on sections of intestinal tissue of uninfected and infected fish revealing the presence of met-enkephalin and serotonin (5-HT) in immuno related cells of the intestine wall. Infected trout had larger numbers of elements positive to met-enkephalin and serotonin antisera. These data provided evidence for the role of the immune system of brown trout in the modulation of the inflammatory response to D. truttae. Results are discussed with respect to host immune response to an intestinal helminth. PMID- 18424173 TI - In vitro effects of fluor-hydroxyapatite, fluorapatite and hydroxyapatite on colony formation, DNA damage and mutagenicity. AB - The number of biomaterials used in biomedical applications has rapidly increased in the past two decades. Fluorapatite (FA) is one of the inorganic constituents of bone or teeth used for hard-tissue repairs and replacements. Fluor hydroxyapatite (FHA) is a new synthetically prepared composite that in its structure contains the same molecular concentration of OH(-) groups and F(-) ions. The aim of this experimental investigation was to evaluate cytotoxic, genotoxic and mutagenic effects of FHA and FA eluates on Chinese hamster V79 cells and to compare them with the effects of hydroxyapatite (HA) eluate. Cytotoxicity of the biomaterials tested was evaluated by use of the cell colony formation assay and by direct counting of the cells in each colony. Genotoxicity was assessed by single-cell gel electrophoresis (comet assay) and mutagenicity was evaluated by the Hprt gene-mutation assay and in bacterial mutagenicity tests using Salmonella typhimurium TA100. The results show that the highest test concentrations of the biomaterials (100% and 75% eluates) induced very weak inhibition of colony growth (about 10%). On the other hand, the reduction of cell number per colony induced by these concentrations was in the range from 43% to 31%. The comet assay showed that biomaterials induced DNA breaks, which increased with increasing test concentrations in the order HAor=65 years), operated in one big general hospital in Athens, were interviewed before, 4 months and 12 months after CABG with the MacNew Heart disease health-related quality of life questionnaire. RESULTS: The majority of the sample were male (N=48, 76.2%), married (N=49, 77.8%) and pensioners (N=54, 61.7%). After the operation and before discharge 42 (66.6%) patients presented complications. One year after the operation, 45 (80.4%) patients experienced improvement and only 11 (19.6%) deterioration in their reported quality of life. Despite this postoperative improvement in all domains, a high percentage of patients (>60%) continued to report exacerbation in questions related to self confidence and dependence to others indicating an overprotective environment. Approximately one in two patients reported signs of cognitive dysfunction during the postoperative period. Elderly patients knew very little about their disease, especially before the operation (mean=2.03, SD=0.69, R=1-5). Educational level, presence of complications in the immediate postoperative period and reported angina were related to a poorer QoL. CONCLUSION: A high proportion of the patients experienced improvement while a substantial number had exacerbations related to self confidence and dependence to others. An important step to improve this situation might be through the institution of a structured multi-disciplinary rehabilitation program with focus on emotional support, information giving and education to elderly CABG patients and their significant others. PMID- 18424184 TI - Age-related changes among 25 patients with congenital cardiac left-to-right shunts and irreversible plexogenic pulmonary arteriopathy. AB - BACKGROUND: Some patients with congenital heart disease and irreversible plexogenic pulmonary arteriopathy survive into adulthood. The purpose of this study was to compare histopathological and antigen expression patterns in adults and children with congenital heart disease and plexogenic pulmonary arteriopathy. METHODS: Autopsy/explant lung tissues from 25 patients with congenital heart disease and plexogenic pulmonary arteriopathy were reviewed for 24 histopathological parameters associated with plexogenic pulmonary arteriopathy, including the prevalence and character of plexiform lesions. Immunohistochemistry using antibodies against CD31, C-kit, smooth muscle actin, CD68, and Fli-1 was performed to evaluate plexiform lesions. RESULTS: Group 1 consisted of 14 patients aged <20 years, and Group 2 included 11 patients aged >or=20 years (range, 20-69 years). Cellular plexiform lesions trended toward greater prevalence in Group 1 than in Group 2 (P=.081), whereas involuted plexiform lesions more frequently affected Group 2 than Group 1 (P=.0037). In addition, recent platelet-fibrin thrombi within plexiform lesions occurred more often in Group 1 than in Group 2 (P=.0419). Intimal proliferation/fibrosis of elastic arteries and pulmonary vein dilation were more common in Group 2 (P=.0172 and P=.0048, respectively). CD31 and C-kit staining in non-lumen-lining cells of plexiform lesions was more frequent in Group 2 than in Group 1 (P=.0858 and P=.0173, respectively). No statistically significant differences in expression patterns between cellular and involuted plexiform lesions existed. CONCLUSIONS: Among patients with congenital heart disease and plexogenic pulmonary arteriopathy, histopathological differences were observed between those surviving into adulthood and those dying in childhood/adolescence. Plexiform lesions in adults undergo remodeling from a cellular morphology to an involuted morphology. It is unclear whether this simply represents the natural progression of plexiform lesions or also confers a survival advantage. PMID- 18424185 TI - ACL-I, a lectin from the marine sponge Axinella corrugata: isolation, characterization and chemotactic activity. AB - The lectin from the marine sponge Axinella corrugata (ACL-I) was purified by affinity chromatography on rabbit erythrocytic stroma incorporated into a polyacrylamide gel followed by gel filtration on Ultrogel AcA 44 column. Purified ACL-I is a hexameric glycoprotein with a Mr of 82.3 kDa estimated by SDS-PAGE and 78.5 kDa by FPLC on Superose 12 HR column. The pI of lectin is 6.3 and ACL-I is constituted of 13.9 kDa similar subunits some of them linked by disulphide bridges. This lectin agglutinates native rabbit, goat and dog erythrocytes and in less extent human erythrocytes. The hemagglutinating activity is independent of Ca(2+), Mg(2+) and Mn(2+), but it is strongly inhibited by carbohydrates containing N-acetyl groups. ACL-I is stable up to 70 degrees C for 30 min, with optimum pH between 7 and 8, and it is also resistant to enzymatic proteolysis in vitro. In the presence of reducing or denaturant agents, the lectin activity decreases. ACL-I displays chemotactic effect on rat neutrophil in vitro which is inhibited by N-acetyl-d-glucosamine. PMID- 18424186 TI - How well do we understand the overall backbone of cycad phylogeny? New insights from a large, multigene plastid data set. PMID- 18424187 TI - An old adaptive radiation of forest dung beetles in Madagascar. AB - Adaptive radiations of mammals have contributed to the exceptionally high levels of biodiversity and endemism in Madagascar. Here we examine the evolutionary history of the endemic dung beetle tribe Helictopleurini (Scarabaeidae) and its relationship to the widely distributed Oniticellini and Onthophagini. Helictopleurini species are dependent on mammals for their resources. We date the single origin of the tribe at 37 to 23 MY ago, indicating overseas colonization of Madagascar. The main radiation occurred concurrently with the main radiations of lemurs. The ancestors of Helictopleurini are inferred to have been coprophagous species inhabiting open habitats. Subsequent evolution has involved a shift into forests, changes in resource use to a more generalized diet, and changes in body size. Four species of the extant 65 species have shifted to use the dung of the recently introduced cattle in open habitats, allowing these species to greatly expand their geographical ranges. PMID- 18424188 TI - Distinct and nonredundant in vivo functions of IFNAR on myeloid cells limit autoimmunity in the central nervous system. AB - The action of type I interferons in the central nervous system (CNS) during autoimmunity is largely unknown. Here, we demonstrate elevated interferon beta concentrations in the CNS, but not blood, of mice with experimental autoimmune encephalomyelitis (EAE), a model for CNS autoimmunity. Furthermore, mice devoid of the broadly expressed type I IFN receptor (IFNAR) developed exacerbated clinical disease accompanied by a markedly higher inflammation, demyelination, and lethality without shifting the T helper 17 (Th17) or Th1 cell immune response. Whereas adoptive transfer of encephalitogenic T cells led to enhanced disease in Ifnar1(-/-) mice, newly created conditional mice with B or T lymphocyte-specific IFNAR ablation showed normal EAE. The engagement of IFNAR on neuroectodermal CNS cells had no protective effect. In contrast, absence of IFNAR on myeloid cells led to severe disease with an enhanced effector phase and increased lethality, indicating a distinct protective function of type I IFNs during autoimmune inflammation of the CNS. PMID- 18424190 TI - The utility of intraluminal impedance in patients with gastroesophageal reflux disease-like symptoms but normal endoscopy and 24-hour pH testing. AB - BACKGROUND & AIMS: Fifty percent of patients with gastroesophageal reflux disease like symptoms and negative endoscopy have negative 24-hour pH testing, suggesting that symptoms are not caused by abnormal esophageal acid exposure. Multichannel intraluminal impedance (MII)-24-hour pH allows the recognition of major acid, minor acid, nonacid, and gas reflux. Recorded symptoms can be correlated with all reflux events (eg, acid, minor acid, nonacid, and gas) and a symptom score can be generated. We aimed to determine whether the Symptom Index (SI) obtained using MII-pH identified an association of symptoms with reflux events in nonclassic acid-reflux disease. METHODS: Thirty-seven patients with heartburn or regurgitation, negative endoscopy, and 24-hour pH were enlisted. Acid suppression was stopped, a 24-hour MII-pH test was performed, and an SI was calculated for major acid reflux alone and for all reflux episodes including major, minor, and nonacid. On this basis patients were divided into 4 groups: (1) standard acid reflux: positive standard pH test; (2) acid sensitive: positive SI for major acid but normal pH test; (3) general reflux: positive SI for major, minor, and nonacid combined, but not for major acid alone; and (4) no reflux: negative SI. RESULTS: Six patients (16%) had standard reflux, 10 patients (27%) had acid-sensitive esophagus, 14 patients (38%) had general reflux, and 7 patients (19%) had a negative SI. CONCLUSIONS: Fifty-seven percent of patients received a diagnosis unachievable with standard pH testing (38% had symptoms associated with general reflux and 19% had no reflux symptom associations). These findings support a potential role for MII-pH testing in this difficult group. PMID- 18424189 TI - Peripheral CD8+ T cell tolerance to self-proteins is regulated proximally at the T cell receptor. AB - CD8(+) T cell tolerance, although essential for preventing autoimmunity, poses substantial obstacles to eliciting immune responses to tumor antigens, which are generally overexpressed normal proteins. Development of effective strategies to overcome tolerance for clinical applications would benefit from elucidation of the immunologic mechanism(s) regulating T cell tolerance to self. To examine how tolerance is maintained in vivo, we engineered dual-T cell receptor (TCR) transgenic mice in which CD8(+) T cells recognize two distinct antigens: a foreign viral-protein and a tolerizing self-tumor protein. Encounter with peripheral self-antigen rendered dual-TCR T cells tolerant to self, but these cells responded normally through the virus-specific TCR. Moreover, proliferation induced by virus rescued function of tolerized self-tumor-reactive TCR, restoring anti-tumor activity. These studies demonstrate that peripheral CD8(+) T cell tolerance to self-proteins can be regulated at the level of the self-reactive TCR complex rather than by central cellular inactivation and suggest an alternate strategy to enhance adoptive T cell immunotherapy. PMID- 18424191 TI - Effect of the antidepressant venlafaxine in functional dyspepsia: a randomized, double-blind, placebo-controlled trial. AB - BACKGROUND & AIMS: Antidepressants could be effective in the treatment of functional gastrointestinal disorders through their anticholinergic and pain modulating effects. Previous studies with these drugs lacked sufficient power and were predominantly conducted in patients with irritable bowel syndrome. This study aimed to assess the effectiveness of the serotonin and norepinephrine reuptake inhibitor venlafaxine in patients with functional dyspepsia. METHODS: This was a multi-center, randomized, double-blind, placebo-controlled trial. Participants had persistent dyspeptic symptoms and underwent upper gastrointestinal endoscopy in a secondary care hospital to exclude organic abnormalities. They were randomly assigned to receive 8 weeks of treatment with either venlafaxine XR (2 weeks 75 mg once daily, 4 weeks 150 mg once daily, and 2 weeks 75 mg once daily) or placebo. Symptoms, health-related quality of life, anxiety, and depression were assessed before and at 4, 8, 12, and 20 weeks after inclusion. RESULTS: One hundred sixty patients were randomized; 56% and 73% of participants completed treatment with venlafaxine or placebo, respectively, according to protocol. There was no difference in proportions of symptom-free patients after 8 weeks of treatment or at 20 weeks after inclusion, with venlafaxine in comparison to placebo (37% and 39%, respectively; odds ratio [OR], 0.8; 95% confidence interval [CI], 0.3-2.1; and 42% and 41%, respectively; OR, 3.1; 95% CI, 0.9-12.6). Per-protocol analysis did not reveal any differences between venlafaxine and placebo either (38% and 39% symptom-free, respectively; OR, 1.0; 95% CI, 0.4-2.4 at 8 weeks). CONCLUSIONS: Treatment with the selective serotonin and norepinephrine reuptake inhibitor venlafaxine is not more effective than placebo in patients with functional dyspepsia. PMID- 18424192 TI - Determination of antimicrobial susceptibilities of clinically isolated anaerobic bacteria by E-test, ATB-ANA and agar dilution. AB - A total of 60 anaerobic strains were isolated from 322 clinical specimens. These isolates were tested for susceptibility to seven antibiotics (penicillin G, amoxicillin/clavulanic acid, cefoxitin, imipenem, chloramphenicol, metronidazole, clindamycin) by using ATB-ANA and Epsilometer test (E-test) strips and the results were compared with the gold standard agar dilution method. Imipenem was found as the most effective agent in vitro among the agents tested (100%). Susceptibility to penicillin G, amoxicillin/clavulanic acid, cefoxitin, chloramphenicol, metronidazole and clindamycin are 36.7%, 83.3%, 88.3%, 96.6%, 85% and 90%, respectively. E-test has showed a good correlation (r=0.62, p=0.001) statistically with the results of agar dilution (total agreement for all antibiotics changing between 90.01% and 98.45%) and a moderate correlation (r=0.45, p=0.048) with the results of ATB-ANA method (total agreement for all antibiotics changing between 75.46% and 98.76%). However, the routine use of agar dilution procedure is concluded to be cumbersome, whereas E-test method offers a reliable alternative. PMID- 18424193 TI - Development of ER-alpha and ER-beta expression in the developing ovine brain and pituitary. AB - Fetal neuroendocrine development in late gestation is critical for maintenance of fetal homeostasis, growth, and readiness for birth. We designed the present study to identify the regional patterns of expression of the two main isoforms of the estrogen receptor, ER-alpha and ER-beta, in the developing ovine fetal brain. Fetal (80, 100, 120, 130, and 145 days gestation), neonatal (1 and 7 days), and adult sheep were euthanized and the following tissues were collected: pituitary, hypothalamus, hippocampus, cerebral cortex, and brainstem. Both ER's are expressed in the ovine brain as early as 80 days gestation, and the expression of both receptors appears to be developmentally regulated. We conclude that both forms of the estrogen receptor are expressed in fetal brain and pituitary throughout the latter half of gestation. PMID- 18424194 TI - Management of atrophic maxilla in severe osteoporosis treated with bisphosphonates: a case report. AB - Studies investigating whether osteoporosis is a risk factor for using implants have revealed no correlation between possible implant failure and the severity of osteoporosis. However, osteoporotic patients frequently require bone regeneration techniques, because they do not fulfil optimum conditions for implant placement owing to the diminished bone structure and rapid resorption of the alveolar bone. We present a successful case of advanced implant therapy using platelet-rich plasma in a patient with severe osteoporosis who had been previously treated with bisphosphonates. We are not aware that this technique has been previously described in literature. PMID- 18424195 TI - Development and validation of a liquid chromatography-atmospheric pressure chemical ionization-tandem mass spectrometry method for simultaneous analysis of 10 amphetamine-, methamphetamine- and 3,4-methylenedioxymethamphetamine-related (MDMA) analytes in human meconium. AB - A liquid chromatography-atmospheric pressure chemical ionization-tandem mass spectrometry (LC-APCI-MS/MS) method for quantification of 10 amphetamine-related analytes in 1g meconium is presented. Specimen preparation included homogenization and solid-phase extraction. Two multiple reaction monitoring transitions were monitored per analyte. Ten and 1 microL injection volumes permitted quantification up to 10,000 ng/g, with sufficient sensitivity to quantify minor metabolites. Lower limits of quantification ranged from 1.25 to 40 ng/g. Precision was less than 14.2%, with accuracy between 79 and 115%. Meconium from a methamphetamine-exposed neonate was analyzed. Metabolites p hydroxymethamphetamine, norephedrine and 4-hydroxy-3-methoxymethamphetamine were identified in meconium for the first time. PMID- 18424196 TI - Disturbed sleep in post-traumatic stress disorder: secondary symptom or core feature? AB - Sleep disturbances are often viewed as a secondary symptom of post-traumatic stress disorder (PTSD), thought to resolve once PTSD has been treated. Specific screening, diagnosis and treatment of sleep disturbances is therefore not commonly conducted in trauma centres. However, recent evidence shows that this view and consequent practices are as much unhelpful as incorrect. Several sleep disorders-nightmares, insomnia, sleep apnoea and periodic limb movements-are highly prevalent in PTSD, and several studies found disturbed sleep to be a risk factor for the subsequent development of PTSD. Moreover, sleep disturbances are a frequent residual complaint after successful PTSD treatment: a finding that applies both to psychological and pharmacological treatment. In contrast, treatment focusing on sleep does alleviate both sleep disturbances and PTSD symptom severity. A growing body of evidence shows that disturbed sleep is more than a secondary symptom of PTSD-it seems to be a core feature. Sleep-focused treatment can be incorporated into any standard PTSD treatment, and PTSD research needs to start including validated sleep measurements in longitudinal epidemiologic and treatment outcome studies. Further clinical and research implications are discussed, and possible mechanisms for the role of disturbed (REM) sleep in PTSD are described. PMID- 18424197 TI - Unsedated transnasal versus transoral sedated upper gastrointestinal endoscopy: a one-series prospective study on safety and patient acceptability. AB - BACKGROUND: While conventional oesophagogastroduodenoscopy is frequently performed under sedation to improve acceptability, transnasal oesophagogastroduodenoscopy would appear to be less invasive. STUDY AIMS: To compare diagnostic accuracy, feasibility, acceptability and safety of transnasal oesophagogastroduodenoscopy without sedation versus conventional oesophagogastroduodenoscopy under sedation. PATIENTS: Following anxiety assessment, 30 dyspeptic patients underwent transnasal oesophagogastroduodenoscopy under local anaesthesia (lidocaine) and conventional oesophagogastroduodenoscopy under conscious sedation (i.v. midazolam) on two consecutive days. Transnasal oesophagogastroduodenoscopy was performed with an ultrathin and conventional oesophagogastroduodenoscopy with a standard endoscope. METHODS: Safety, evaluated by monitoring cardio-respiratory functions. Acceptability, rated according to discomfort and preference between the two examinations. Diagnostic accuracy evaluated taking into account endoscopic patterns and adequacy of biopsy specimens for histology. Feasibility, defined according to endoscopic performance, quality of images and overall opinion of the endoscopist. Only gastric biopsies were evaluated. RESULTS: All patients but one who refused conventional oesophagogastroduodenoscopy underwent both transnasal oesophagogastroduodenoscopy and conventional oesophagogastroduodenoscopy. No cardiorespiratory complications occurred during either technique. Majority of patients (87%) preferred transnasal oesophagogastroduodenoscopy. Examinations were completed in all cases, with comparable endoscopic patterns. All biopsy specimens were suitable for histology. CONCLUSIONS: Transnasal oesophagogastroduodenoscopy without sedation provides good diagnostic accuracy, is safer and better accepted than conventional oesophagogastroduodenoscopy under sedation and, therefore, represents a valid alternative in routine diagnosis of upper digestive tract diseases. PMID- 18424198 TI - High-speed cinematographic evaluation of claw-ground contact pattern of lactating cows. AB - To evaluate the manner in which a cow's claws make contact with the ground at the walk, the gait, and in particular the claw-ground contact pattern, were studied in 12 healthy, lactating dairy cows, using high-speed cinematography (500frames/s) while the animals were walking on a treadmill. The results showed that the limbs were advanced around the contralateral limbs in a sigmoid curve. The feet contacted the ground with the foot axis and the tips of the claws rotated slightly outwards. In all cows the lateral claws contacted the ground before the medial claws in the hindlimbs, and in 10/12 cows in the forelimbs. The heel of the lateral claws was the region of initial contact with the ground in the hindlimbs of all cows and in the forelimbs in 9/12 cows. Lateral 'heel first' contact in the fore and hindlimbs appeared to be the normal gait pattern in these animals. Compared with a previous study of heifers, lactating cows had a larger step width in the hindlimbs and a smaller step width in the forelimbs. These ground contact patterns offer an explanation for the predisposition to claw disorders of the lateral claw of the hindlimb. The results of this study reinforce the suggestion that soft floor surfaces should be provided for cattle to prevent mechanical injury to the claws. PMID- 18424199 TI - Modelling responses to nutritional, endocrine and genetic strategies to increase fertility in the UK dairy herd. AB - The United Kingdom, as in most countries using intensive dairy management programmes, is facing serious challenges in terms of dairy cow fertility, as highlighted by a rapidly increasing calving interval (CI). A mechanistic, mathematical model is described that predicts the size of the future national dairy herd required to supply domestic requirements and its inherent sustainability in terms of production of replacement female numbers. The results from the model suggest that continuing use of current management strategies may result in the national dairy herd being unsustainable due to increasing CI and reduced fertility in as few as 10years. Adoption of nutritional, endocrine and genetic techniques that increase fertility can effectively and rapidly reverse this trend and reduce the required size of the national herd, thereby reducing methane emissions from dairy production. PMID- 18424200 TI - Effect of intraoperative fluid optimisation on renal function in patients undergoing emergency abdominal surgery: a randomised controlled pilot study (ISRCTN 11799696). AB - BACKGROUND: Emergency abdominal surgery carries a high risk of postoperative morbidity and mortality. Goal directed therapy has been advocated to improve outcome in high-risk surgery. The aim of the present pilot study was to examine the effect of goal directed therapy using fluid alone on postoperative renal function and organ failure score in patients undergoing emergency abdominal surgery. METHODS: This prospective randomised pilot study included patients over the age of 50 undergoing emergency abdominal surgery. In the intervention group pulse pressure variation measurements were used to guide fluid boluses of 6% Hydroxyethylstarch 130/0.4. The control group received standard care. Serum urea, creatinine and cystatin C levels were measured prior to and at the end of surgery and postoperatively on day 1, day 3 and day 5. RESULTS: Thirty patients were recruited. One patient died prior to surgery and was excluded from the analysis. The intervention group received a median of 750 ml of hydroxyethylstarch. The peak values of postoperative urea were 6.9 (2.7-31.8) vs. 6.4 (3.5-11.5)mmol/l (p=0.425), creatinine 100 (60-300) vs. 85 (65-150) micromol/l (p=0.085) and cystatin C 1.09 (0.66-4.94) vs. 1.01 (0.33-2.29)mg/dl (p=0.352) in the control and intervention group, respectively. CONCLUSIONS: In the present pilot study replacing the identified fluid deficit was not associated with a change in renal function. These results do not preclude that goal directed therapy using fluid alone may have an effect on renal function but they would suggest that the effect size of fluid optimisation alone on renal function is small. PMID- 18424201 TI - Encephalitis lethargica in 5 South African children. AB - The clinical features and cognitive outcome in 5 South African childhood cases of sporadic encephalitis lethargica seen between 2002 and 2006 are discussed. All children presented with an acute encephalopathic illness complicated by sleep disturbance, extrapyramidal and neuropsychiatric symptoms. Diagnosis was based on shared clinical features with other cases described in the literature and exclusion of known infective, biochemical and metabolic causes of acute childhood encephalopathy. The negative findings on neuro-imaging in all cases strongly supported the diagnosis. All children survived but 3 cases became learning disabled and all required cognitive rehabilitation after recovery. The cases demonstrate that encephalitis lethargica does indeed occur among South African children. The condition should be considered in any previously well child that presents with an acute encephalopathic illness with prominent extrapyramidal and neuropsychiatric symptoms and negative infectious, biochemical, autoimmune, metabolic and radiologic investigations. Recognition is important as it allows counseling of parents regarding the protracted course but generally favorable outcome of the condition. PMID- 18424203 TI - Pulse sequence for dynamic volumetric imaging of hyperpolarized metabolic products. AB - Dynamic nuclear polarization and dissolution of a (13)C-labeled substrate enables the dynamic imaging of cellular metabolism. Spectroscopic information is typically acquired, making the acquisition of dynamic volumetric data a challenge. To enable rapid volumetric imaging, a spectral-spatial excitation pulse was designed to excite a single line of the carbon spectrum. With only a single resonance present in the signal, an echo-planar readout trajectory could be used to resolve spatial information, giving full volume coverage of 32 x 32 x 16 voxels every 3.5s. This high frame rate was used to measure the different lactate dynamics in different tissues in a normal rat model and a mouse model of prostate cancer. PMID- 18424205 TI - Parallel, stochastic measurement of molecular surface area. AB - Biochemists often wish to compute surface areas of proteins. A variety of algorithms have been developed for this task, but they are designed for traditional single-processor architectures. The current trend in computer hardware is towards increasingly parallel architectures for which these algorithms are not well suited. We describe a parallel, stochastic algorithm for molecular surface area computation that maps well to the emerging multi-core architectures. Our algorithm is also progressive, providing a rough estimate of surface area immediately and refining this estimate as time goes on. Furthermore, the algorithm generates points on the molecular surface which can be used for point-based rendering. We demonstrate a GPU implementation of our algorithm and show that it compares favorably with several existing molecular surface computation programs, giving fast estimates of the molecular surface area with good accuracy. PMID- 18424204 TI - Cytogenetic and molecular characterization of a de-novo t(2p;7p) translocation involving TNS3 and EXOC6B genes in a boy with a complex syndromic phenotype. AB - We describe a premature newborn child with left renal agenesis, right low functional kidney, altered chemical-clinical parameters, neutropenia, recurrent pulmonary infections, long bone diaphysis broadening, growth and developmental delay. Postnatal cytogenetic analysis revealed a 46,XY,t(2;7)(p13;p12) de-novo karyotype. The chromosome breakpoints were defined by FISH using BAC probes and initially restricted to about 123,000bp in 2p13 and delimited to 84,600bp in 7p12. Bioinformatic analysis of these genomic regions showed two genes that are involved in the rearrangement: exocyst C6B (EXOC6B) for chromosome 2 breakpoint and tensin3 (TNS3) for chromosome 7 breakpoint. A EXOC6B-TNS3 fusion transcript together with a reciprocal TNS3-EXOC6B chimeric RNA have been detected by RT-PCR performed on skin fibroblasts RNA of the proband. These data localize the chromosome 2 breakpoint within the first intron of EXOC6B, while the translocation event on chromosome 7 occurred in intron 15 of TNS3. We hypothesize that the phenotype observed in the patient results from one or several mechanisms including: haploinsufficiency of EXOC6B and TNS3 genes; a dominant negative effect exerted by the chimeric transcripts; a disregulation in the expression of other genes adjacent the breakpoints. Although no clear evidences exist supporting a role of any of the above mentioned mechanisms in the pathogenesis of the complex phenotype, immunofluorescence analysis of tensin1 in the patient's fibroblasts suggests that the TNS3 gene haploinsufficiency results in a reduced expression of tensin1. These cells may be therefore a model for understanding the role and the organization of the tensin protein family. PMID- 18424206 TI - Effects of food abundance, age, and flea infestation on the body condition and immunological variables of a rodent host, and their consequences for flea survival. AB - Temporal variation in body condition and immunological variables of animals that harbor parasites may explain patterns of variation in infestation, as well as parasite impact on the host. We emulated such variability in Sundevall's jirds by manipulating food availability and flea infestation in juveniles and adults and examining how these changes affect survival of fleas on their hosts. Body condition of food-restricted jirds deteriorated, but there was no change in their immunological variables. Adult jirds were in better body condition and had higher immunocompetence than juveniles, however there were no significant effects of flea infestation on any of the variables examined. The main effects of flea infestation were a decrease in the response to phytohaemagglutinin injection, and an increase in the negative effects of food restriction on body mass. Flea survival was higher on juveniles, but fleas did not respond to temporal variability in body condition and immunocompetence of the jirds. We concluded that changes in body condition and immune responses due to growth or variability in food abundance are more important than changes caused by the fleas themselves. Flea infestation is more detrimental to jirds when they are not able to compensate for mass loss through increased food consumption. PMID- 18424207 TI - Minimal volume regulation after shrinkage of red blood cells from five species of reptiles. AB - Red blood cells (RBCs) from most vertebrates restore volume upon hypertonic shrinkage and the mechanisms underlying this regulatory volume increase (RVI) have been studied extensively in these cells. Despite the phylogenetically interesting position of reptiles, very little is known about their red cell function. The present study demonstrates that oxygenated RBCs in all major groups of reptiles exhibit no or a very reduced RVI upon approximately 25% calculated hyperosmotic shrinkage. Thus, RBCs from the snakes Crotalus durissus and Python regius, the turtle Trachemys scripta and the alligator Alligator mississippiensis showed no statistically significant RVI within 120 min after shrinkage, while the lizard Tupinambis merianae showed 22% volume recovery after 120 min. Amiloride (10(-4) M) and bumetanide (10(-5) M) had no effect on the RVI in T. merianae, indicating no involvement of the Na(+)/H(+) exchanger (NHE) or the Na(+)/K(+)/2Cl(-) co-transporter (NKCC) or insentive transporters. Deoxygenation of RBCs from A. mississippiensis and T. merianae did not significantly affect RVI upon shrinkage. Deoxygenation per se of red blood cells from T. merianae elicited a slow volume increase, but the mechanism was not characterized. It seems, therefore, that the RVI response based on NHE activation was lost among the early sauropsids that gave rise to modern reptiles and birds, while it was retained in mammals. An RVI response has then reappeared in birds, but based on activation of the NKCC. Alternatively, the absence of the RVI response may represent the most ancient condition, and could have evolved several times within vertebrates. PMID- 18424208 TI - Developmental changes and the impact of isoflavones on mRNA expression of IGF-I receptor, EGF receptor and related growth factors in porcine skeletal muscle cell cultures. AB - OBJECTIVE: Soy that is widely used in human nutrition and in livestock production is a rich source of isoflavones. In addition to the estrogenic or antiestrogenic effects, isoflavones are suggested to affect cell growth via inhibition of protein tyrosine kinases (e.g. growth factor receptors). Therefore, the present in vitro-study was undertaken to determine, whether genistein and daidzein affect the mRNA expression of growth factor receptors (IGF-I receptor and EGF receptor) and their related growth factors in porcine skeletal muscle cell cultures. DESIGN: First, we investigated the basal mRNA expression of IGF-I, IGF-II, EGF, IGF-I receptor, and EGF receptor in proliferating and differentiating porcine skeletal muscle cell cultures using real-time PCR. Secondly, we measured the changes in the mRNA expression in these cell cultures treated with 0 (control), 1, 10, 100 microM genistein or daidzein over 26 h in serum-free medium (n=3). RESULTS: The mRNA expression of IGF-I was slightly decreased, whereas transcript concentrations of IGF-II and EGF were increased during differentiation compared with the proliferating stage of porcine muscle cell cultures. IGF-I receptor transcripts tended to be increased, whereas EGF receptor mRNA expression remained unchanged from proliferation to differentiation. Genistein and daidzein at 1 microM and 10 microM showed no effects on the mRNA expression of these genes, neither in proliferating nor in differentiating cells. However, high-concentrated isoflavones (100 microM) decreased the mRNA expression of IGF-I receptor and of the growth factors examined. CONCLUSIONS: The present study confirms the role of the IGF and EGF system in proliferation and differentiation of skeletal muscle cell culture especially under serum-free culture conditions. Furthermore, the results of this in vitro-study suggest that there is no effect of isoflavones at concentrations resulting from dietary consumption (1 and 10 microM) on IGF- and EGF-associated gene expression in porcine skeletal muscle tissue. Genistein and daidzein at high concentration (100 microM) reduced the mRNA expression of the IGF-I receptor and the growth factors examined, and therefore, may modify their autocrine and paracrine actions in skeletal muscle tissue. PMID- 18424210 TI - OPHDIAT: quality-assurance programme plan and performance of the network. AB - AIMS: There is a need for evaluation of screening and grading services for diabetic retinopathy (DR) in compliance with quality-assurance (QA) standards. We describe the screening/grading QA programme set up for OPHDIAT over the 2005-2006 period. METHODS: Screening and grading objectives, evaluation criteria and minimum acceptable QA standards were set. To ensure the quality of DR photos, the proportion of nongradable photos in at least one eye had to be less than 10%. To ensure grading accuracy, intergrading agreement had to be greater than 90%. Grader-generated reports had to be available in less than 48 h for more than 80% photos. Readers had to grade 500 to 3000 photos per year. RESULTS: Sixteen screening centres were opened between June 2004 and December 2006, and 14,769 patients were screened. Percentages of nongradable photos were consistently below the QA requirement (less than 10%). Overall, 800 photos were graded a second time by a reader blinded to original grading; agreement between graders ranged from 92 to 99%. More than 90% of grader-generated reports were produced within 48 h. The number of readings by each grader nearly achieved the QA standard. CONCLUSION: QA for DR telescreening should be a continuous process to provide performance feedback, thus guaranteeing a high standard for delivered results. Almost all of the predetermined QA standards in OPHDIAT for screening and grading were met. Besides the quality/sensitivity of the screening/grading modalities, it is important to evaluate at-risk patients so that they can be treated efficiently; this should be addressed in a global QA programme. PMID- 18424209 TI - A three-dimensional model of a group II intron RNA and its interaction with the intron-encoded reverse transcriptase. AB - Group II introns are self-splicing ribozymes believed to be the ancestors of spliceosomal introns. Many group II introns encode reverse transcriptases that promote both RNA splicing and intron mobility to new genomic sites. Here we used a circular permutation and crosslinking method to establish 16 intramolecular distance relationships within the mobile Lactococcus lactis Ll.LtrB-DeltaORF intron. Using these new constraints together with 13 established tertiary interactions and eight published crosslinks, we modeled a complete three dimensional structure of the intron. We also used the circular permutation strategy to map RNA-protein interaction sites through fluorescence quenching and crosslinking assays. Our model provides a comprehensive structural framework for understanding the function of group II ribozymes, their natural structural variations, and the mechanisms by which the intron-encoded protein promotes RNA splicing and intron mobility. The model also suggests an arrangement of active site elements that may be conserved in the spliceosome. PMID- 18424211 TI - Organizing pneumonia after rituximab therapy: Two cases. AB - Rituximab, a chimeric monoclonal antibody against CD20, very rarely causes lung toxicity. Clinical presentations include lung infiltrates, alveolar hemorrhage, and adult respiratory distress syndrome. Three cases of organizing pneumoinia (formerly called bronchiolitis obliterans with organizing pneumonia or BOOP) have been reported. In our experience, organizing pneumonia occurred in 2 of 25 patients treated with rituximab, for RA and Castleman's disease, respectively. Because organizing pneumonia may be asymptomatic, as illustrated by one of our cases, we recommend obtaining a chest radiograph routinely before rituximab re treatment. PMID- 18424212 TI - Selecting first-line treatment for patients with osteoporosis. PMID- 18424213 TI - [Role of homeopathy in gynecology]. PMID- 18424214 TI - [How I perform... a laparoscopic cervicoisthmic cerclage]. PMID- 18424215 TI - [Information in perinatal medicine: ethical aspects]. PMID- 18424216 TI - [Male breast cancer: a review of 52 cases collected at the Institute Bergonie (Bordeaux, France) from 1980 to 2004]. AB - OBJECTIVES: To analyze the characteristics and to establish prognosis factors for 52 men suffering from breast cancer from 1980 to 2004. PATIENTS AND METHODS: Men treated for breast cancer (invasive or in situ). A retrospective study analyzed clinical and histological characteristics, and treatment procedures. The probability of survival or recurrence was calculated using the Kaplan-Meier method. Prognostic factors were studied using the Log Rank test. RESULTS: The mean age of our patients was 63.5 years old. In 73.1% of cases, subaerolar tumors were the initial symptoms, the average size was 30.31 mm. Among patients, 17 (32.7%) had T1, 19 (36.5%) T2, two (3.8%) T3 and 14 (26.9%) T4. The most represented histological type was the infiltrative ductal carcinoma (84.6%). The spread rate to axillary lymph nodes was 63.6%. The hormone dependency of these tumors was proven in 84.6% of cases. Overall survival rate were about 69% at five years and 32% at 10 years. The spread to lymph node and to derm, the clinical stage were significant factors influencing disease free survival. None of these factors had any significance regarding overall survival. DISCUSSION AND CONCLUSION: Male breast cancer is a rare disease (about 1% of breast cancer) with a poor prognosis (32% 10 years disease free survival). An early diagnosis and better knowledge of the disease would certainly lead to improvement of prognosis. PMID- 18424217 TI - [Rupture of non-communicating rudimentary pregnant uterine horn in a pseudo unicornuate uterus at 23 weeks of amenorrhea. Case report]. AB - Pregnancy in the rudimentary horn of a unicornuate uterus is an extremely rare form of ectopic gestation associated with a high risk of uterine rupture. We report the case of a pregnancy developed in a non communicating rudimentary horn of a unicornuate uterus complicated by horn rupture at 23 weeks of amenorrhea showing as an acute abdominal pain and massive hemoperitoneum. This patient's uterine abnormality was known before, as this woman has delivered two years before at term a healthy boy by cesarean section. This past pregnancy was located in the normal horn and the non communicating rudimentary horn seemed at this time normal. This uterine malformation is presented with its gynecological and obstetrical entailments as well as methods that could prevent such outcome. PMID- 18424218 TI - Protein release from yeast cells as an evaluation method of physical effects in ultrasonic field. AB - The release rate of intercellular protein from yeast cells by the ultrasonic action is proposed as a method for evaluating the physical (mechanical) effects of the ultrasonic field. The protein concentration was quantitatively determined using UV absorbance of proteins by spectrophotometry. The detail of the procedures, such as the effects of the origin of yeast cells, pretreatment of the cells, and the wavelengths for spectrophotometric determination of protein content, are examined. The effectiveness of the proposed evaluation method was experimentally demonstrated by changing the irradiation conditions of ultrasound, such as the concentration of yeast cells, temperature, ultrasound power, types of sonicator, and the superposition with the mechanical mixing. The results validate the usefulness of the proposed evaluation method for the quantification of the physical effects of ultrasound fields. Also, the range of cavitational effects of ultrasound sensed by the evaluation procedures were discussed. PMID- 18424219 TI - An efficient and practical synthesis of mandelic acid by combination of complex phase transfer catalyst and ultrasonic irradiation. AB - Mandelic acid was synthesized from benzaldehyde with chloroform in a 81% yield at 60 degrees C for 2 h by using triethylbenzylammonium chloride (TEBA) and poly(ethylene glycol)-800 (PEG-800) as a complex phase transfer catalyst under ultrasonic irradiation. The main advantages of this present method are that the reaction time is much shorter and the yield is higher than those of the classical method. p-Methoxymandelic acid was also obtained in a 84% yield under the same reaction conditions. PMID- 18424220 TI - From 1997 to 2007: a decade journey through the H1 haplotype on 17q21 chromosome. AB - The H1 haplotype was first identified 10 years ago. Initially, a dinucleotide polymorphism was detected in the tau (MAPT) gene and was subsequently found to be in linkage disequilibrium (LD) with other polymorphisms, forming the MAPT H1 haplotype, a risk factor for many neurological diseases, considered as tauopathies. Genetic and histopathologic data are in agreement that MAPT and its encoded protein have a pivotal role in the normal function of neurons. Currently, the H1 haplotype extends beyond the outer edges of MAPT encompassing multiple genes on chromosome 17 and thus increasing the number of candidate genes implicated in the pathogenesis of tauopathies. This review highlights the milestones and basic events in the journey towards uncovering the significance of the H1 haplotype. PMID- 18424222 TI - Post-translational regulation of plant bZIP factors. AB - The post-translational regulation of transcription factors plays an important role in the control of gene expression in eukaryotes. The mechanisms of regulation include not only factor modifications but also regulated protein protein interaction, protein degradation and intracellular partitioning. In plants, the basic-region leucine zipper (bZIP) transcription factors contribute to many transcriptional response pathways. Despite this, little is known about their post-translational regulation. Recent findings suggest that plant bZIP factors are under the control of various partially signal-induced and reversible post-translational mechanisms that are crucial for the control of their function. However, the fact that, to date, only a few plant bZIPs have been analyzed with respect to post-translational regulation indicates that we have just identified the tip of an iceberg. PMID- 18424221 TI - Learning and consolidation of visuo-motor adaptation in Parkinson's disease. AB - We have previously shown in normal subjects that motor adaptation to imposed visual rotation is significantly enhanced when tested few days later. This occurs through a process of sleep-dependent memory consolidation. Here we ascertained whether patients with Parkinson's disease (PD) learn, improve, and retain new motor skills in the same way as normal subjects. We tested 16 patients in early stages of PD and 21 control subjects over two days. All subjects performed reaching movements on a digitizing tablet. Vision of the limb was precluded with an opaque screen; hand paths were shown on the screen with the targets' position. Unbeknownst to the subjects, the hand path on the screen was rotated by 30 degrees . In experiment 1, patients taking dopaminergic treatment and controls adapted to rotation with targets appearing in an unpredictable order. In experiment 2, drug-naive patients and controls adapted to rotation in a less challenging task where target's appearance was predictable. Patients and controls made similar movements and adapted to rotation in the same way. However, when tested again over the following days, controls' performance significantly improved compared to training, while patients' performance did not. This lack of consolidation, which is present in the early stages of the disease and is independent from therapy, may be due to abnormal homeostatic processes that occur during sleep. PMID- 18424223 TI - Molecular mechanisms of floral mimicry in orchids. AB - Deceptive plants do not produce floral rewards, but attract pollinators by mimicking signals of other organisms, such as food plants or female insects. Such floral mimicry is particularly common in orchids, in which flower morphology, coloration and odour play key roles in deceiving pollinators. A better understanding of the molecular bases for these traits should provide new insights into the occurrence, mechanisms and evolutionary consequences of floral mimicry. It should also reveal the molecular bases of pollinator-attracting signals, in addition to providing strategies for manipulating insect behaviour in general. Here, we review data on the molecular bases for traits involved in floral mimicry, and we describe methodological advances helpful for the functional evaluation of key genes. PMID- 18424224 TI - Does the chimpanzee have a theory of mind? 30 years later. AB - On the 30th anniversary of Premack and Woodruff's seminal paper asking whether chimpanzees have a theory of mind, we review recent evidence that suggests in many respects they do, whereas in other respects they might not. Specifically, there is solid evidence from several different experimental paradigms that chimpanzees understand the goals and intentions of others, as well as the perception and knowledge of others. Nevertheless, despite several seemingly valid attempts, there is currently no evidence that chimpanzees understand false beliefs. Our conclusion for the moment is, thus, that chimpanzees understand others in terms of a perception-goal psychology, as opposed to a full-fledged, human-like belief-desire psychology. PMID- 18424225 TI - Patterns of prescribing extended wear contact lenses. AB - Patterns of prescribing extended wear contact lenses in the UK were determined by mining through data gathered from annual contact lens fitting surveys conducted over the past 12 years. The increased rate of extended wear prescribing this century - which in 2007 had reached 7% and 19% of all soft lens new fits and refits, respectively - probably reflects the superior clinical performance of silicone hydrogel lenses for this purpose. PMID- 18424226 TI - Genotoxic effects of white fluorescent light on human lymphocytes in vitro. AB - Sources of light beams such as white fluorescent light, are present in our daily life to meet the needs of life in the modern world. This study was conducted with the objective of determining the possible genotoxic, cytotoxic and aneugenic effects caused by this agent in different stages of the cell cycle (G0/early G1, S, and late G2), using different cytogenetic parameters (sister chromatid exchanges--SCE, chromosome aberrations--CA, and detection of aneugenic effects) in lymphocytes from temporary cultures of human peripheral blood. WFL showed a genotoxic effect in vitro, expressed by an increase in the frequency of SCE's, regardless of the cell cycle stage. However, no increase in the frequency of CAs was observed. In addition, disturbances in cell cycle kinetics and chromosomal segregation were also observed. Taken together, such data may contribute to a better understanding and a different management in the use of phototherapy for some pathological conditions. PMID- 18424227 TI - Rad52 has a role in the repair of sodium selenite-induced DNA damage in Saccharomyces cerevisiae. AB - Selenium (Se) is a chemo-preventive agent that has been shown to have a protective role against cancer. The inorganic form of Se, sodium selenite (Na2SeO3), has frequently been included in various chemo-prevention studies, and this commercially available form of Se is used as dietary supplement by the public. Because high doses of this Se compound can be toxic, the underlying molecular mechanisms of sodium selenite toxicity need to be elucidated. Recently, we have reported that sodium selenite is acting as an oxidizing agent in the budding yeast Saccharomyces cerevisiae, producing oxidative damage to DNA. This pro-oxidative activity of sodium selenite likely accounted for the observed DNA double-strand breaks (DSB) and yeast cell death. In this study we determine the genetic factors that are responsible for repair of sodium selenite-induced DSB. We report that the Rad52 protein is indispensable for repairing sodium selenite induced DSB, suggesting a fundamental role of homologous recombination (HR) in this repair process. These results provide the first evidence that HR may have a fundamental role in the repair of sodium selenite-induced toxic DNA lesions. PMID- 18424228 TI - An investigation on electron transfer quenching of zinc(II) meso tetraphenylporphyrin (ZnTPP) by colloidal TiO(2). AB - The interaction of zinc(II) meso-tetraphenylporphyrin (ZnTPP) with colloidal TiO(2) was studied by absorption, steady state and time-resolved fluorescence spectroscopy. The quenching was found to obey the Stern-Volmer equation and the corresponding Stern-Volmer plots were linear in the range of quencher concentration used 0-5 x 10(-4)M. The bimolecular quenching rate constants (k(q)) were 20.5 x 10(10)M(-1)s(-1) (steady-state) and 2.85 x 10(10)M(-1)s(-1) (time resolved). The quenching process is suggested to involve electron transfer from the ZnTPP to TiO(2) considering the experimental evidences obtained. PMID- 18424229 TI - Type-specific reproducibility of the Roche linear array HPV genotyping test. AB - BACKGROUND: Type-specific detection of human papillomavirus (HPV) is increasingly important for monitoring temporal and age-specific changes in type-specific prevalence in support of HPV vaccination efforts. The impact of sampling, extraction and assay characteristics on HPV results is increasingly recognized. Inter-assay comparability studies have been performed, but the robustness of type specific results has neither been emphasized nor has the degree of intra-assay reproducibility been addressed. OBJECTIVES: Here we describe the general and type specific reproducibility of the linear array HPV genotyping test (Roche Molecular Diagnostics, Indianapolis, IN). STUDY DESIGN: Extracts of 276 cervical samples from two ongoing epidemiologic HPV studies were retested while blinded to prior results. The testing involved five different reagent lots and three technologists. RESULTS: Concordance for HPV detection (sample positive versus negative for any of the 37 types) was high (98.2%, kappa=0.959). Type-specific concordance for individual HPV types was also high (99.4%, kappa=0.915), and most samples (83.0%) showed complete concordance for all types. CONCLUSIONS: Type specific reproducibility of the linear array HPV genotyping test is good but not perfect. The results suggest that type-specific performance should be included in the evaluation of HPV typing formats. PMID- 18424230 TI - Establishing correlations of scalp field maps with other experimental variables using covariance analysis and resampling methods. AB - OBJECTIVE: In EEG/MEG experiments, increasing the number of sensors improves the spatial resolution of the results. However, the standard statistical methods are inappropriate for these multivariate, highly correlated datasets. We introduce a procedure to identify spatially extended scalp fields that correlate with some external, continuous measure (reaction-time, performance, clinical status) and to test their significance. METHODS: We formally deduce that the channel-wise covariance of some experimental variable with scalp field data directly represents intracerebral sources associated with that variable. We furthermore show how the significance of such a representation can be tested with resampling techniques. RESULTS: Simulations showed that depending on the number of channels and subjects, effects can be detected already at low signal to noise ratios. In a sample analysis of real data, we found that foreign-language evoked ERP data were significantly associated with foreign-language proficiency. Inverse solutions of the extracted covariances pointed to sources in language-related areas. CONCLUSIONS: Covariance mapping combined with bootstrapping methods has high statistical power and yields unique and directly interpretable results. SIGNIFICANCE: The introduced methodology overcomes some of the 'traditional' statistical problems in EEG/MEG scalp data analysis. Its application can improve the reproducibility of results in the field of EEG/MEG. PMID- 18424232 TI - Immunocytochemical localization of serotonin in the central and peripheral chemosensory system of mosquitoes. AB - Female mosquitoes depend on blood to complete their reproductive cycle and rely mainly on chemosensory systems to obtain blood meals. An immunocytochemical analysis reveals a number of serotonin-immunoreactive neurons that innervate the chemosensory systems, suggesting a potential role of serotonin in modulating chemosensory processes. In the primary olfactory system, we identify a single ipsilateral centrifugal neuron with arborizations in higher brain centers; the varicosities of this neuron display volumetric changes in response to both blood feeding and during a circadian rhythm. Six to eight pairs of serotonin immunoreactive neurons are identified in the primary gustatory neuropil, including the subesophageal ganglion and tritocerebrum. The peripheral chemosensory organs, i.e. the antenna, the maxillary palp and the labium, are described as having extensive serotonergic neurohemal plexi. In addition, we describe the presence of serotonin-immunoreactive fibers in the mechanosensory Johnston's organ. Taking these results together, we discuss the potential role of serotonin as a neuromodulator in the chemosensory system of disease vector mosquitoes. PMID- 18424234 TI - Recent advances in pulmonary trichomonosis. PMID- 18424233 TI - Prostate-cancer mortality in the USA and UK in 1975-2004: an ecological study. AB - BACKGROUND: There is no conclusive evidence that screening based on serum prostate-specific antigen (PSA) tests decreases prostate-cancer mortality. Since its introduction in the USA around 1990, uptake of PSA testing has been rapid in the USA, but much less common in the UK. Our aim was to study trends over time in prostate-cancer mortality and incidence in the USA and UK in 1975-2004, and compare these patterns with trends in screening and treatment. METHODS: Joinpoint regression analysis of cancer-mortality statistics from Cancer Research UK (London, UK) and from the US National Cancer Institute Surveillance, Epidemiology and End Results (SEER) programme from 1975 to 2004 was used to estimate the annual percentage change in prostate-cancer mortality in both countries and the points in time when trends changed. The ratio of USA to UK age-adjusted prostate cancer incidence was also assessed. FINDINGS: Age-specific and age-adjusted prostate-cancer mortality peaked in the early 1990s at almost identical rates in both countries, but age-adjusted mortality in the USA subsequently declined after 1994 by -4.17% (95% CI -4.34 to -3.99) each year, four-times the rate of decline in the UK after 1992 (-1.14% [-1.44 to -0.84]). The mortality decline in the USA was greatest and most sustained in patients aged 75 years or older (-5.32% [-8.23 to -2.32]), whereas death rates had plateaued in this age group in the UK by 2000. The mean ratio of USA to UK age-adjusted prostate-cancer incidence rates in 1975-2003 was 2.5, with a pronounced peak around the time that PSA testing was introduced in the USA. Numbers needed to treat to prevent one death from prostate cancer were 33 000 in the 55-64-year age group. INTERPRETATION: The striking decline in prostate-cancer mortality in the USA compared with the UK in 1994-2004 coincided with much higher uptake of PSA screening in the USA. Explanations for the different trends in mortality include the possibility of an early effect of initial screening rounds on men with more aggressive asymptomatic disease in the USA, different approaches to treatment in the two countries, and bias related to the misattribution of cause of death. Speculation over the role of screening will continue until evidence from randomised controlled trials is published. PMID- 18424235 TI - Ecological immunology of mosquito-malaria interactions. AB - More than a century after the discovery of the complex life cycle of its causative agent, malaria remains a major health problem. Understanding mosquito malaria interactions could lead to breakthroughs in malaria control. Novel strategies, such as the design of transgenic mosquitoes refractory to Plasmodium, or design of human vaccines emulating mosquito resistance to the parasite, require extensive knowledge of processes involved in immune responses and of microevolutionary mechanisms that create and maintain variation in immune responses in wild vector populations. The recent realization of how intimately and specifically mosquitoes and Plasmodium co-evolve in Nature is driving vector molecular biologists and evolutionary ecologists to move closer to the natural setting under the common umbrella of 'Ecological immunology'. PMID- 18424236 TI - Visilizumab induces apoptosis of mucosal T lymphocytes in ulcerative colitis through activation of caspase 3 and 8 dependent pathways. AB - Visilizumab, a humanized low-Fc receptor binding anti-CD3 antibody, induces rapid clinical response in patients with steroid-refractory ulcerative colitis (UC). Several effective treatments in IBD have been linked to the induction of mucosal T cell apoptosis. The aim of the present study was to evaluate the effect of visilizumab on the apoptosis of lamina propria (LP) and peripheral blood (PB) lymphocytes isolated from patients with UC. Visilizumab induced dose- and time dependent apoptosis of LP T cells isolated from non-IBD individuals, UC or CD patients. Maximal effect was seen at a concentration of 100 ng/ml and it was 33% for normal, 34% for UC and 23% for CD LP T cells following 24 h stimulation. Visilizumab induced apoptosis predominantly of CD4(+) LP T cells, whereas CD8(+) LP T cells were relatively resistant to apoptosis. Visilizumab did not induce apoptosis of PB T cells from UC patients. Visilizumab-induced apoptosis of LP T cells was dependent on caspase 3 and 8, but not caspase 9 activation and did not involve the Fas/FasL pathway. Low-Fc receptor binding Abs such as visilizumab may be highly effective for the treatment of UC through induction of apoptosis of LP T cells and rapid elimination of lesional pathogenic T cells in the gut mucosa. PMID- 18424237 TI - Prevalence, patterns, service needs, and assessment of neuropsychiatric disorders among people with epilepsy in residential care: validation of the Neuropsychiatric Inventory as a caregiver-rated measure of neuropsychiatric functioning in epilepsy. AB - We assessed 228 people with epilepsy (PWE) in the residential care setting using the Neuropsychiatric Inventory (NPI) and Brief Psychiatric Rating Scale (BPRS) as caregiver- and observer-rated instruments. There was a significant burden of psychopathology, about half of all subjects surveyed scoring positive on either or both instruments. Psychopathology as measured by the NPI and BPRS was significantly greater in cognitively impaired subjects than in those with intact cognitive function. The NPI was found to be a valid caregiver-rated measure of psychopathology in PWE, with a principal components analysis yielding a reliable and interpretable four-factor solution, psychosis, interictal dysphoric disorder, depression, and anxiety being identified. Mental health service needs were found to be considerable in this population, with a significant hidden burden of psychiatric comorbidity. As this population has ongoing service needs through the life span, further research is necessary. PMID- 18424238 TI - Transcriptional effects of PFOS in isolated hepatocytes from Atlantic salmon Salmo salar L. AB - The main aim of the current in vitro experiment was to search for makers for PFOS exposure in isolated hepatocytes from Atlantic salmon Salmo salar, based on genes responding to PFOS exposure in other animals. Primary cell cultures of hepatocytes were exposed to four concentrations of PFOS (2.1-6.2-15.1-25.0 mg/L) for 24 and 48 h and the transcriptional levels of 12 genes encoding proteins known to respond to PFOS were quantified with real-time RT-PCR. The 12 examined genes were caspase 3B (apoptosis), GSH-Px and HSP70 (cellular stress), CYP1A, CYP3A, GST and UGT (P450 and phase II enzymes), acyl-CoA oxidase, PPARalpha, PPARbeta and PPARgamma (lipid metabolism) and Na(+)-K(+)-ATPase (ion regulation). Most of the studied genes responded in a dose-dependent manner to PFOS exposure, although the transcriptional differences in general where small with regard to fold change. Our results clearly suggest that PFOS exposure enhanced cellular stress in the examined cells, even though the exact mechanisms behind this stress remain unknown. The results from this in vitro experiment showed that genes known to be affected by PFOS exposure in other species also were induced in hepatocytes of Atlantic salmon, giving us the rationale to expand to testing the actual in vivo magnitude of effect in Atlantic salmon exposed to PFOS at doses usually seen in nature/diets. PMID- 18424239 TI - The Impact of CAROtid plaque Screening on Smoking (CAROSS) cessation and control of other cardiovascular risk factors: Rationale and design of a randomized controlled trial. AB - BACKGROUND: Screening tests for subclinical cardiovascular disease, such as markers of atherosclerosis, are increasingly used in clinical prevention to identify individuals at high cardiovascular risk. Being aware of these test results might also enhance patient motivation to change unhealthy behaviors but the effectiveness of such a screening strategy has been poorly studied. METHODS: The CAROtid plaque Screening trial on Smoking cessation (CAROSS) is a randomized controlled trial in 530 regular smokers aged 40-70 years to test the hypothesis that carotid plaque screening will influence smokers' behavior with an increased rate of smoking cessation (primary outcome) and an improved control of other cardiovascular risk factors (secondary outcomes) after 1-year follow-up. All smokers will receive a brief advice for smoking cessation,and will subsequently be randomly assigned to either the intervention group (with plaques screening) or the control group (without plaque screening). Carotid ultrasound will be conducted with a standard protocol. Smokers with at least one carotid plaque will receive pictures of their own plaques with a structured explanation on the general significance of plaques. To ensure equal contact conditions, smokers not undergoing ultrasound and those without plaque will receive a relevant explanation on the risks associated with tobacco smoking. Study outcomes will be compared between smokers randomized to plaque screening and smokers not submitted to plaque screening. SUMMARY: This will be the first trial to assess the impact of carotid plaque screening on 1-year smoking cessation rates and levels of control of other cardiovascular risk factors. PMID- 18424240 TI - Electrode commutation sequence for honeycomb arrangement of electrodes in electrochemotherapy and corresponding electric field distribution. AB - Electrochemotherapy is a treatment based on combination of chemotherapeutic drug and electroporation. It is used in clinics for treatment of solid tumours. For electrochemotherapy of larger tumours multiple needle electrodes were already suggested. We developed and tested electrode commutation circuit, which controls up to 19 electrodes independently. Each electrode can be in one of three possible states: on positive or negative potential or in the state of high impedance. In addition, we tested a pulse sequence using seven electrodes for which we also calculated electric field distribution in tumour tissue by means of finite elements method. Electrochemotherapy, performed by multiple needle electrodes and tested pulse sequence on large subcutaneous murine tumour model resulted in tumour growth delay and 57% complete responses, thus demonstrating that the tested electrode commutation sequence is efficient. PMID- 18424241 TI - Na+-glucose cotransporter is also expressed in mesothelium of species with thick visceral pleura. AB - Molecular evidence for Na+-glucose cotransporter (SGLT1) in rabbit pleural mesothelium has been recently provided, confirming earlier functional findings on solute-coupled liquid absorption from rabbit pleural space. In this research we checked whether SGLT1 is also expressed in pleural mesothelium of species with thick visceral pleura, which receives blood from systemic circulation, but drains it into pulmonary veins. To this end immunoblot assays were performed on total protein extract of scraped visceral and parietal mesothelium of lambs and adult sheep, and of a human mesothelial cell line. All of them showed SGLT1 specific bands. Moreover, confocal immunofluorescence images of lamb pleural mesothelium showed that SGLT1 is located in apical membrane. Therefore, a solute-coupled liquid absorption should also occur from pleural space of species with thick visceral pleura. Because of this protein-free liquid entering interstitium between visceral mesothelium and capillaries, inherent Starling forces should be different than hitherto considered, and visceral pleura capillaries could absorb liquid even in these species. PMID- 18424242 TI - Identification of the heparin-binding domain of TNF-alpha and its use for efficient TNF-alpha purification by heparin-Sepharose affinity chromatography. AB - The N-terminus of the trimeric TNF-alpha molecule comprises two basic arginines within the short amino-acid sequence VRSSSR, which is here shown to be essential for binding of TNF-alpha to heparin-Sepharose. Mixed trimers containing full length and DeltaN6-truncated subunits revealed a single VRSSSR sequence to be sufficient to achieve binding. On the basis of this newly identified heparin binding domain, a new method for efficient purification of TNF-alpha is described. Affinity chromatography on heparin-Sepharose was introduced as a key step for highly purified TNF-alpha at a high yield. With minor modifications, this procedure can be used for TNF-alpha analogues that have full-length N termini, as shown for the less toxic analogue LK-805. PMID- 18424243 TI - Gastric epithelial dysplasia: the Western perspective. AB - The need for early diagnosis of gastric cancer is emphasized by the fact that gastric cancer remains the second most common cause of cancer related deaths worldwide. The aggressive surveillance and definite therapy for low and high grade dysplasia, which can be achieved endoscopic means, remains the cornerstone of clinical management. Although the precursor status of dysplasia is not contested, its classification is controversial and fraught with marked inter observer variations. Most cases of gastric dysplasia have an "intestinal" phenotype referred to as adenomatous dysplasia. Hyperplastic (type II dysplasia) is another less common variant. The progression of dysplasia to carcinoma is paralleled by a stepwise accumulation of multiple, but yet uncertain, genetic abnormalities. There are no immunohistochemical or molecular assays that can stratify with certainty the risk of progression to cancer. Given the low rate of transformation of low-grade dysplasia, annual endoscopic surveillance with re biopsy is advocated. A diagnosis of indefinite for dysplasia should also prompt endoscopic surveillance. A diagnosis of high-grade dysplasia is more ominous, since it progress to cancer in most cases. However, the novel imaging and endoscopic modalities have modified management strategies with mucosal lesions amenable to endoscopic resection, while surgical resection is reserved to invasive adenocarcinoma with submucosal invasion. PMID- 18424245 TI - Impacts of exposure to images of ideal bodies on male body dissatisfaction: a review. AB - Research suggests that young men's body dissatisfaction increases when they see images of attractive muscular men. This article provides the first extensive review of experimental studies exposing men to advertisements or commercials featuring idealized male bodies. Impacts on body dissatisfaction were evaluated by calculating and analyzing effect sizes from 15 studies. The effect sizes indicate that exposure to images of idealized male bodies has a small but statistically significant negative impact on men's body dissatisfaction. Three studies suggest that young men who are dissatisfied with their bodies are at increased risk for negative self-evaluations when exposed to idealized images. Two studies suggest that men who are satisfied with their bodies may be protected against negative impacts from seeing such images. PMID- 18424244 TI - OLGA staging for gastritis: a tutorial. AB - Atrophic gastritis (resulting mainly from long-standing Helicobacter pylori infection) is a major risk factor for (intestinal-type) gastric cancer development and the extent/topography of the atrophic changes significantly correlates with the degree of cancer risk. The current format for histology reporting in cases of gastritis fails to establish an immediate link between gastritis phenotype and risk of malignancy. The histology report consequently does not give clinical practitioners and gastroenterologists an explicit message of use in orienting an individual patient's clinical management. Building on current knowledge of the biology of gastritis and incorporating experience gained worldwide by applying the Sydney System for more than 15 years, an international group of pathologists (Operative Link for Gastritis Assessment) has proposed a system for reporting gastritis in terms of stage (the OLGA staging system). Gastritis staging arranges the histological phenotypes of gastritis along a scale of progressively increasing gastric cancer risk, from the lowest (stage 0) to the highest (stage IV). This tutorial aims to provide unequivocal information on how to consistently apply the OLGA staging system in routine diagnostic histology practice. PMID- 18424246 TI - Neonatal polycythemia and hyperviscosity. AB - Neonatal polycythemia and hyperviscosity are defined as a hematocrit > or =65% and a viscosity value >2 standard deviations greater than the norm. Although polycythemia can reflect normal fetal adaptation, it has been thought to be responsible for abnormalities in the neonate. Polycythemia and hyperviscosity are associated with blood-flow changes in some organs, which alter their function. Partial exchange transfusion (PET) has been used to treat both symptomatic and asymptomatic patients. At present, no data support the use of PET in asymptomatic infants; the potential benefit in symptomatic infants depends on the symptoms. Studies of long-term neurodevelopmental status do not show any clear long-term benefits for PET. Crystalloids are as effective as colloids in PET and have the advantage of being cheaper and more readily available; also, they do not confer any risk of infection or anaphylaxis. PMID- 18424247 TI - In vivo perfusion analysis of normal and dysplastic ears and its implication on total auricular reconstruction. AB - INTRODUCTION: During the first stage of total auricular reconstruction with autologous rib cartilage, according to Nagata, the cartilage framework is placed in a subcutaneous pocket. Its posterior skin flap is relatively large and can be prepared with or without a subcutaneous pedicle. This represents the crucial part of the procedure, as impaired healing and infection can occur due to low perfusion. Nothing was known about the blood supply of ear remnants or flap perfusion during reconstruction. It was not clear whether the preservation of the additional subcutaneous pedicle secures high blood supply. METHODS: We used laser fluorescence angiography with indocyanine green dye for anatomical and functional perfusion studies in eight normal ears. Subsequently the anatomical and functional vessel architecture of 18 dysplastic ears was investigated. Finally, five patients each were operated on with or without subcutaneous pedicle during auricular reconstruction and intraoperatively monitored with laser fluorescence angiography. RESULTS: We showed that the vessel structure of normal ears detected by fluorescence angiography is equivalent to anatomical preparations. The surrounding skin in high grade microtia remnants is biphasically perfused by deep perforators and by the cutaneous vessel network. The preservation of the subcutaneous pedicle during auricular reconstruction leads to significantly better perfusion of the posterior skin flap. No signs of critical perfusion or complications were observed in these patients. DISCUSSION: We present the feasibility of laser fluorescence angiography to simultaneously gain anatomical and functional data about skin blood supply. The first anatomical and functional description of blood supply of ear remnants in third grade microtia is given. Functional data of skin flap perfusion during and after complete ear reconstruction were evaluated. The present study shows that the subcutaneous pedicle of Nagata's procedure is of great importance for success of the first stage operation as it prevents impaired wound healing. In contrast, patients without a subcutaneous pedicle had a broad spectrum of reduced perfusion and therefore some had complications. PMID- 18424248 TI - The alteration of a mechanical property of bone cells during the process of changing from osteoblasts to osteocytes. AB - Osteocytes acquire their stellate shape during the process of changing from osteoblasts in bone. Throughout this process, dynamic cytoskeletal changes occur. In general, changes of the cytoskeleton affect cellular mechanical properties. Mechanical properties of living cells are connected with their biological functions and physiological processes. In this study, we for the first time analyzed elastic modulus, a mechanical property of bone cells. Bone cells in embryonic chick calvariae and in isolated culture were identified using fluorescently labeled phalloidin and OB7.3, a chick osteocyte-specific monoclonal antibody, and then observed by confocal laser scanning microscopy. The elastic modulus of living cells was analyzed with atomic force microscopy. To examine the consequences of focal adhesion formation on the elastic modulus, cells were pretreated with GRGDS and GRGES, and then the elastic modulus of the cells was analyzed. Focal adhesions in the cells were visualized by immunofluorescence of vinculin. From fluorescence images, we could distinguish osteoblasts, osteoid osteocytes and mature osteocytes both in vivo and in vitro. The elastic modulus of peripheral regions of cells in all three populations was significantly higher than in their nuclear regions. The elastic modulus of the peripheral region of osteoblasts was 12053+/-934 Pa, that of osteoid osteocytes was 7971+/-422 Pa and that of mature osteocytes was 4471+/-198 Pa. These results suggest that the level of elastic modulus of bone cells was proportional to the stage of changing from osteoblasts to osteocytes. The focal adhesion area of osteoblasts was significantly higher than that of osteocytes. The focal adhesion area of osteoblasts was decreased after treatment with GRGDS, however, that of osteocytes was not. The elastic modulus of osteoblasts and osteoid osteocytes were decreased after treatment with GRGDS. However, that of mature osteocytes was not changed. There were dynamic changes in the mechanical property of elastic modulus and in focal adhesions of bone cells. PMID- 18424249 TI - Development of in vivo muCT evaluation of neovascularisation in tissue engineered bone constructs. AB - Due to an increasing aging population the need for innovative approaches to aid skeletal repair and reconstruction is a significant socio-economic increasing problem. The emerging discipline of tissue engineering has sort to augment the growth and repair of bone loss particularly in areas of trauma, degeneration and revision surgery. However, the initiation and development of a fully functional vascular network are critical for bioengineered bone to repair large osseous defects, whether the material is osteosynthetic (poly (d,l)-lactic acid, PLA) or natural bone allograft. Quantification and three-dimensional visualization of new vessel networks remain a problem in bone tissue engineering constructs. A novel technique utilising a radio-opaque dye and micro-computed tomography (muCT) has been developed and applied to study angiogenesis in an impaction bone graft model. Tissue-engineered constructs combining human bone marrow stromal cells (HBMSC) with natural allograft and synthetic grafts (PLA) were impacted and implanted into the subcutis of MF-1 nu/nu mice for a period of 28 days. Microfil consisting of radio-opaque polymer was perfused through the mice and scanned using a Bench Top CT system for micro-computed tomography. Analysis of three dimensional muCT reconstructions demonstrated an increase in vessel volume and vessel number in the impacted scaffolds/HBMC compared to scaffolds alone. Vessel volume: allograft/HBMSC=0.57 mm(3)+/-0.19; allograft=0.04 mm(3)+/-0.04; PLA/HBMSC=1.19 mm(3)+/-0.31; and PLA=0.12 mm(3)+/-0.01. Penetrating vessel number: allograft/HBMSC=22.33+/-3.21; allograft=3.67+/-1.153; PLA/HBMSC=32.67+/ 8.33; and PLA=7.67+/-3.06. Type 1 collagen and von Willebrand factor immunohistochemistry in scaffold/HBMSC constructs indicated the osteogenic cell phenotype, and new blood vessel formation respectively. Contrast-enhanced 3D reconstructions facilitated the visualization and quantification of neovascularisation. This novel technique has been used to demonstrate neovascularisation in impacted tissue engineered constructs providing a facile approach with wide experimental application. PMID- 18424251 TI - Quantitative gene expression analysis in a brain slice model: influence of temperature and incubation media. AB - We describe the RNA integrity (28S/18S ratio) and the messenger RNA (mRNA) expression of genes encoding glyceraldehyde 3-phosphate dehydrogenase (GAPDH), microtubule-associated serine/threonine kinase 2 (Mast2), and beta-actin in cortical brain slices incubated for up to 24h in Ringer's solution and Dulbecco's modified Eagle's medium (DMEM) at 25 and 37 degrees C. Our data reveal an optimal temporal working window between 1 and 6h when slices are incubated in Ringer's solution at 25 degrees C that allows experiments related to gene expression dynamics to be performed more suitably than those carried out at 37 degrees C. In addition, we show that reference gene expression may be modified in dynamic experiments and may compromise studies of gene expression. PMID- 18424252 TI - A new method for the mapping of 5' ends of RNAs. AB - In this article, we describe a new procedure to map 5' ends of RNAs. The procedure consists in the use of specific RNase H digestion of a hybrid formed by the RNA and a complementary DNA oligonucleotide. Northern blot hybridization of the resulting RNA fragment allows an accurate measurement of its length. Although we generally use this procedure as a control of previously performed primer extension analyses, the absence of nonspecific bands, which often occur in primer extensions on RNA templates with extended secondary structures, suggests that our method may be preferable when these difficult templates are analyzed. PMID- 18424253 TI - De-phosphorylation of GR at Ser203 in nuclei associates with GR nuclear translocation and GLUT5 gene expression in Caco-2 cells. AB - Glucocorticoid hormones and p44/42 mitogen-activated protein kinase (MAPK) inactivation are considered to be important in small-intestinal differentiation/maturation. In this study, we found that co-treatment with glucocorticoid hormone agonist dexamethasone and p44/42 MAPK inhibitor PD98059 in intestinal cell line Caco-2 strongly induced GLUT5 gene expression. Glucocorticoid hormone receptor (GR) was translocated from the cytoplasm to the nucleus and de-phosphorylated at serine residue 203 in the nucleus, by combined treatment with dexamethasone and PD98059. The binding of GR, as well as acetylated histones H3 and H4, to the promoter/enhancer region of GLUT5 gene was enhanced by combined treatment with dexamethasone and PD98059. These results suggest that the inactivation of p44/42 MAP kinase enhances glucocorticoid hormone-induced GLUT5 gene expression, probably through controlling the phosphorylation at serine 203 and nuclear transport of GR, as well as histone acetylation on the promoter/enhancer region of GLUT5 gene. PMID- 18424254 TI - The direct role of vitamin D on bone homeostasis. AB - Vitamin D requires two metabolic conversions, 25-hydroxylation in the liver and 1alpha-hydroxylation in the kidney, before its hormonal form, 1,25 dihydroxyvitamin D [1,25-(OH)2D], can bind to the vitamin D receptor (VDR) to modulate gene transcription and regulate mineral ion homeostasis. The receptor and metabolic enzymes are expressed in many tissues, however, which has long suggested that the vitamin D hormone could act in an autocrine, paracrine, or intracrine fashion to affect the biology of non-classical target tissues. Strong support for this model has been obtained for Toll-like receptor-mediated innate immunity in macrophages, for example. The classical view is that vitamin D exerts its effects on bone indirectly via control of calcium and phosphate homeostasis, despite expression of cyp27b1, the 25-hydroxyvitamin D-1alpha-hydroxylase, and the VDR in osteoblasts and chondrocytes. Recent molecular genetic studies have revealed direct, but non-essential roles for 1,25-(OH)2D in growth plate chondrocytes. Specific inactivation of the VDR in collagen type II-expressing chondrocytes leads to reduced RANKL expression and delayed osteoclastogenesis, which causes a transient increase in bone volume at the primary spongiosa. Chondrocyte-specific VDR-ablated mice also show reduced circulating levels of FGF23 and thus elevated serum phosphate concentrations. The mechanisms remain to be completely determined but appear to involve a 1,25-(OH)2D-induced secreted factor from chondrocytes that affects FGF23 production by neighboring osteoblasts. The phenotype of additional mutant mice models, including chondrocyte-specific inactivation or overexpression of cyp27b1, is being analyzed to provide further support for these results that show autocrine and paracrine roles for 1,25-(OH)2D during endochondral bone development. PMID- 18424256 TI - The osteocyte lineage. AB - The osteocyte resides in the lacuna/canalicular system in bone and has been hypothesized to orchestrate local bone remodeling. Certainly the identification of the osteocyte as the source of Sclerostin, a molecule that regulates osteoblast function, has supported this possibility. As our understanding of this cell increases it has become clear that it has more far reaching influence than simply local bone turnover activity. The osteocyte is also the source of DMP-1 and FGF-23, the later being a hormone that regulates kidney function in terms of phosphate uptake. We now see the osteocyte as having important roles both locally in the skeleton and also in other distant tissues. The study of osteocyte biology has reached a particularly exiting level of maturity and illustrates the value of this cell type as a drug discovery target. PMID- 18424255 TI - Cell-cell communication in the osteoblast/osteocyte lineage. AB - Skeletal development (bone modeling) and its maintenance in post-natal life in response to local and systemic stimuli (bone remodeling) require coordinated activity among osteoblasts (bone forming cells), osteocytes (cells embedded in bone) and osteoclasts (bone resorbing cells), in order to meet the needs of structural integrity, mechanical competence and maintenance of mineral homeostasis. One mechanism of cell-cell interaction is via direct cell-cell communication via gap junctions. These are transmembrane channels that allow continuity of cytoplasms between communicating cells. The biologic importance of connexin43 (Cx43), the most abundant gap junction protein in the skeleton is demonstrated by the skeletal malformations present in oculodentodigital dysplasia (ODDD), a disease linked to Cx43 gene (GJA1) mutations, and by the low bone mass and osteoblast dysfunction in Gja1 ablated mice. The presence of Cx43 is required for osteoblast differentiation and function, and by forming either gap junctions or "hemichannels" Cx43 allows participation of cell networks to responses to extracellular stimuli, via propagation of specific signals converging upon connexin sensitive transcriptional units. Hence, Cx43 is involved in skeletal responsiveness to anabolic signals, as those provided by parathyroid hormone and physical load, the latter function probably involving osteocyte-osteoblast communication. PMID- 18424257 TI - (-)-Epigallocatechin gallate induces Nrf2-mediated antioxidant enzyme expression via activation of PI3K and ERK in human mammary epithelial cells. AB - The chemopreventive and chemoprotective activities of green tea have been attributed to the polyphenolic ingredient (-)-epigallocatechin-3-gallate (EGCG). Here, we report that treatment of human breast epithelial (MCF10A) cells with EGCG induces the expression of glutamate-cysteine ligase, manganese superoxide dismutase (MnSOD), and heme oxygenase-1 (HO-1). NF-E2-related factor (Nrf2) has been reported to regulate the antioxidant response element (ARE)-mediated expression of many antioxidant as well as detoxifying enzymes. The nuclear accumulation, ARE binding and transcriptional activity of Nrf2 were increased by EGCG treatment. Silencing of Nrf2 by siRNA gene knockdown rendered the MCF10A cells less sensitive to the EGCG-induced expression of HO-1 and MnSOD. Furthermore, EGCG activated Akt and extracellular signal-regulated protein kinase1/2 (ERK1/2). The pharmacologic inhibition of these kinases abrogated the nuclear translocation of Nrf2 induced by EGCG. These findings suggest that Nrf2 mediates EGCG-induced expression of some representative antioxidant enzymes, possibly via Akt and ERK1/2 signaling, which may provide the cells with acquired antioxidant defense capacity to survive the oxidative stress. PMID- 18424258 TI - Osteoclast lineage and function. AB - Osteoclasts are members of the monocyte/macrophage lineage and are formed by cellular fusions from their mononuclear precursors. Their differentiation is regulated by a number of other cells and their products, especially by RANKL and M-CSF. The resorbing osteoclasts are polarized and show specific plasma membrane domains. Polarization and bone resorption need a continuous membrane trafficking and modulation of the cytoskeleton. The most characteristic feature of osteoclasts is their unique capacity to dissolve crystalline hydroxyapatite by targeted secretion of HCl into the extracellular resorption lacuna. Organic matrix is degraded by enzymes like cathepsin K and the degradation products are transcytosed through the cell for secretion. Dissolution of hydroxyapatite releases large amounts of soluble calcium, phosphate and bicarbonate. Removal of these ions apparently involves the vesicular pathways and direct ion transport via different ion exchangers, channels and pumps. Detailed molecular knowledge of osteoclast differentiation and function has helped us to identify several target molecules and develop specific treatments to inhibit pathological bone resorption in various skeletal diseases. PMID- 18424259 TI - A dynamic model of nitric oxide inhibition of mitochondrial cytochrome c oxidase. AB - Nitric oxide can inhibit mitochondrial cytochrome oxidase in both oxygen competitive and uncompetitive modes. A previous model described these interactions assuming equilibrium binding to the reduced and oxidised enzyme respectively (Mason, et al. Proc. Natl. Acad. Sci. U S A 103 (2006) 708-713). Here we demonstrate that the equilibrium assumption is inappropriate as it requires unfeasibly high association constants for NO to the oxidised enzyme. Instead we develop a model which explicitly includes NO binding and its enzyme bound conversion to nitrite. Removal of the nitrite complex requires electron transfer to the binuclear centre from haem a. This revised model fits the inhibition constants at any value of substrate concentration (ferrocytochrome c or oxygen). It predicts that the inhibited steady state should be a mixture of the reduced haem nitrosyl complex and the oxidized-nitrite complex. Unlike the previous model, binding to the oxidase is always proportional to the degree of inhibition of oxygen consumption. The model is consistent with data and models from a recent paper suggesting that the primary effect of NO binding to the oxidised enzyme is to convert NO to nitrite, rather than to inhibit enzyme activity (Antunes et al. Antioxid. Redox Signal. 9 (2007) 1569-1579). PMID- 18424260 TI - The pre-transmembrane region of the HCV E1 envelope glycoprotein: interaction with model membranes. AB - The previously identified membranotropic regions of the HCV E1 envelope glycoprotein, a class II membrane fusion protein, permitted us to identify different sequences which might be implicated in viral membrane fusion, membrane interaction and/or protein-protein binding. HCV E1 glycoprotein presents a membrano-active region immediately adjacent to the transmembrane segment, which could be involved in membrane destabilization similarly to the pre-transmembrane domains of class I fusion proteins. Consequently, we have carried out a study of the binding and interaction with the lipid bilayer of a peptide corresponding to segment 309-340, peptide E1PTM, as well as the structural changes which take place in both the peptide and the phospholipid molecules induced by the binding of the peptide to the membrane. Here we demonstrate that peptide E1(PTM) strongly partitions into phospholipid membranes, interacts with negatively-charged phospholipids and locates in a shallow position in the membrane. These data support its role in HCV-mediated membrane fusion and suggest that the mechanism of membrane fusion elicited by class I and II fusion proteins might be similar. PMID- 18424261 TI - Quantifying the effects of co-expressing EGFR and HER2 on HER activation and trafficking. AB - The human epidermal growth factor receptor (HER) system is an intricately regulated system that plays critical roles in development and tumorigenesis. Here, we apply integrated experimentation and modeling to analyze HER receptor activation in a panel of cell lines expressing endogenous levels of EGFR/HER1 and different levels of HER2. A mathematical model that includes the fundamental processes involved in receptor activation and trafficking was used to fit the experimental data, and values of the independent parameters for active receptor dimer formation affinities, trafficking rates and relative phosphorylation levels were estimated. Obtained parameter values quantitatively support the existing ideas on the effect of HER2 on EGFR dynamics, and enable us to predict the abundances of various phosphorylated receptor dimers in the cell lines. PMID- 18424262 TI - Transcription of the caspase-14 gene in human epidermal keratinocytes requires AP 1 and NFkappaB. AB - Caspase-14, a protease involved in skin barrier formation, is specifically expressed in epidermal keratinocytes (KCs). Here, we mapped three start sites of transcription of the human caspase-14 gene and analyzed the upstream chromosomal region for promoter activity. Reporter gene assays identified a core promoter region proximal to the first exon and a distal regulatory region which differentially suppressed promoter activity in KC and other cells. Sequence elements in the proximal promoter were bound by the transcription factors AP-1 (JunB, c-Jun, JunD, Fra-1 and Fra-2) and NFkappaB (p50 and RelB). Our data reveal the basic organization of the human caspase-14 promoter and suggest an important role of AP-1 and NFkappaB in the transcriptional control of caspase-14. PMID- 18424263 TI - Single-amino-acid mutation in the HA alters the recognition of H9N2 influenza virus by a monoclonal antibody. AB - We explored the molecular basis of antigenic variation by comparing two H9N2 subtype avian influenza viruses, A/Chicken/Shandong/6/96 (CK/SD/6) and A/Chicken/Guangxi/10/99 (CK/GX/10), that react differently to a monoclonal antibody C/B3. To assess the genetic basis for this antigenic difference, we used reverse genetics to generate a series of chimera and mutants of these two viruses. We found that a single-amino-acid substitution of asparagine for serine at position 145 (S145N) in the HA protein prevents the reaction of CK/SD/6 virus with C/B3. Substitution of serine for asparagine at the same position (N145S) enables the CK/GX/10 to react with C/B3 in hemaglutinin inhibition, immunofluorescence and neutralization assays. We further demonstrated that the amino acid N145 in the H9 HA protein is glycosylated. Our results provide experimental evidence that the glycosylation of HA oligosaccharide attachment sites implicated in antibody binding could have a role in antigenic variation. PMID- 18424264 TI - Importance of SARS-CoV spike protein Trp-rich region in viral infectivity. AB - SARS-CoV entry is mediated by spike glycoprotein. During the viral and host cellular membrane fusion, HR1 and HR2 form 6-helix bundle, positioning the fusion peptide closely to the C-terminal region of ectodomain to drive apposition and subsequent membrane fusion. Connecting to the HR2 region is a Trp-rich region which is absolutely conserved in members of coronaviruses. To investigate the importance of Trp-rich region in SARS-CoV entry, we produced different mutated S proteins using Alanine scan strategy. SARS-CoV pseudotyped with mutated S protein was used to measure viral infectivity. To restore the aromaticity of Ala-mutants, we performed rescue experiments using phenylalanine substitutions. Our results show that individually substituted Ala-mutants substantially decrease infectivity by >90%, global Ala-mutants totally abrogated infectivity. In contrast, Phe substituted mutants are able to restore 10-25% infectivity comparing to the wild type. The results suggest that the Trp-rich region of S protein is essential for SARS-CoV infectivity. PMID- 18424265 TI - Identification of tumor antigens that elicit a humoral immune response in breast cancer patients' sera by serological proteome analysis (SERPA). AB - BACKGROUND: In this study we applied a serological proteomics-based approach (SERPA) to identify tumor antigens that commonly induce a humoral immune response in patients with infiltrating ductal breast carcinomas. METHODS: Sera obtained at the time of diagnosis from 40 patients with invasive breast cancer and 42 healthy controls were screened individually for the presence of IgG antibodies to MCF-7 cell line proteins. Immunoreactive proteins were isolated and subsequently identified by MALDI-TOF mass spectrometry. RESULTS: We identified 26 proteins that reacted with antibodies in the sera from breast cancer patients. Among these antigens, a significantly higher frequency occurs against the molecular chaperone HSP60, the tumor suppressor prohibitin, beta-tubulin, the haptoglobin-related protein and peroxiredoxin-2. Immunoreactivity to hnRNPK, Mn-SOD and F1-ATPase was also clearly detected in the patients group, whereas scarcely in control sera. By contrast, two other antigens identified as cytokeratins 8 and 18, as well as, F1 actin were found to elicit humoral immune responses in both control and breast cancer patients' sera. CONCLUSIONS: The immunoproteomic approach implemented here offers a powerful tool for determining novel tumor antigens that elicit a humoral immune response in patients with invasive breast cancer. These antigens and/or their related circulating antibodies may display clinical usefulness as potential diagnostic markers and provide a means for a better understanding of the molecular mechanisms underlying breast cancer development. PMID- 18424266 TI - Undergraduate education in Laboratory Medicine. AB - The graduation of medicine is a title necessary but not sufficient to practice the medical profession. In Italy, as in other countries, Laboratory Medicine teaching is an educational area integrated with relevant pre-clinical disciplines, supporting the main medical processes of patient diagnosis, prognosis, treatment monitoring, evolution following-up, risk screening and hidden/latent condition finding. In the modern concept of core curriculum of medical students, some cardinal aspects of Laboratory Medicine should be included and precisely: test ordering, preanalytical and analytical issues, test interpreting. Such learning objectives would ultimately contribute, by a procedural/methodological approach, to the process of either correctly diagnosing or proper decision making, at the patient's benefit. The present article provides an overview of Laboratory Medicine teaching in the core curriculum of medical students in Italy, and highlights specific tasks of laboratory competencies and skills required to optimize the learning abilities of the students. PMID- 18424267 TI - Airway ion transport impacts on disease presentation and severity in cystic fibrosis. AB - OBJECTIVES: Abnormal airway ion transport is a feature of cystic fibrosis. The aim of this study was to investigate whether distinct components of ion transport are associated with the clinical expression and severity of the disease. DESIGN AND METHODS: Univariate and multivariate analyses were used to study interaction effects between nasal potential difference parameters and clinical status, recorded at stable conditions, in 75 F508del homozygous young adults. RESULTS: All patients demonstrated increased sodium and reduced chloride conductances. Less sodium transport abnormalities were related to better respiratory function and nutrition. Presentation with digestive symptoms at diagnosis was associated with lower chloride conductance. With an accuracy of 85% good nutritional status was linked to more preserved lung function, increasing age and more preserved chloride conductance. CONCLUSIONS: Ion transport abnormalities have distinct clinical outcomes. Sodium conductance relates to respiratory function and nutrition; chloride conductance to nutrition and presentation with digestive symptoms at diagnosis. PMID- 18424268 TI - Redirection of regulatory T cells with predetermined specificity for the treatment of experimental colitis in mice. AB - BACKGROUND & AIMS: Treatment with ex vivo expanded regulatory T cells (Tregs) is regarded as a promising therapeutic approach in inflammatory bowel disease but is hampered by impaired Treg accumulation and function at inflammatory sites. We aim to study whether antigen-specific redirected Tregs can overcome these limitations. METHODS: We developed transgenic mice whose T cells, including Tregs, express chimeric receptor (CR) made of antibody variable region as recognition unit and T-cell stimulatory and costimulatory domains to activate specifically in response to the predetermined model antigen 2,4,6-trinitrophenol (TNP). RESULTS: TNP-specific CR-bearing Tregs were potently and specifically activated by exogenous TNP and suppressed effector T cells in the absence of costimulatory B7-CD28 interaction. TNP-specific transgenic (Tg) mice were resistant to 2,4,6-trinitrobenzene sulphonic acid (TNBS) colitis but not to other hapten-mediated colitis. Adoptive transfer of CR-bearing Tregs to wild-type mice with TNBS colitis was associated with significant amelioration of colitis and improved survival. Although TNP-specific CR-bearing Tregs did not suppress oxazolone colitis, they cured it after addition of traces of TNBS to oxazolone inflamed colons, demonstrating a "bystander" effect. In vivo imaging of adoptively transferred CR-bearing Tregs demonstrated that they preferentially migrate to TNBS-induced colonic mucosal lesions within hours of induction of colitis. CONCLUSIONS: Tregs can be redirected with specificity distinct from that of pathogenic lymphocytes, accumulate at colonic inflammatory lesions, and suppress effector T cells in a specific, nonmajor histocompatibility complex restricted, and noncostimulatory-dependent manner, resulting in significant amelioration of colitis. Hopefully, this approach will lead to a novel therapy for inflammatory bowel disease, as well as other inflammatory diseases. PMID- 18424270 TI - Cationic liposomal lipids: from gene carriers to cell signaling. AB - Cationic lipids are positively charged amphiphilic molecules which, for most of them, form positively charged liposomes, sometimes in combination with a neutral helper lipid. Such liposomes are mainly used as efficient DNA, RNA or protein carriers for gene therapy or immunization trials. Over the past decade, significant progress has been made in the understanding of the cellular pathways and mechanisms involved in lipoplex-mediated gene transfection but the interaction of cationic lipids with cell components and the consequences of such an interaction on cell physiology remains poorly described. The data reported in the present review provide evidence that cationic lipids are not just carriers for molecular delivery into cells but do modify cellular pathways and stimulate immune or anti-inflammatory responses. Considering the wide number of cationic lipids currently available and the variety of cellular components that could be involved, it is likely that only a few cationic lipid-dependent functions have been identified so far. PMID- 18424269 TI - Hepatocellular carcinoma is associated with an increased risk of hepatitis B virus recurrence after liver transplantation. AB - BACKGROUND & AIMS: Hepatitis B virus (HBV) recurrence after orthotopic liver transplantation (OLT) is significantly reduced by prophylaxis with hyperimmune antibody to hepatitis B surface antigen (anti-HBs) globulins (HBIG) and antiviral drugs. The role of hepatocellular carcinoma (HCC) in HBV recurrence remains unclear. We investigated the association between HCC pre-OLT and HBV recurrence post-OLT. METHODS: We studied 99 hepatitis B surface antigen-positive patients who underwent OLT for cirrhosis. The median follow-up period was 43 months. All patients received HBIG, and 51 also received lamivudine and/or adefovir. Of these 99 patients, 31 had HCC before OLT. Total HBV DNA and covalently closed circular (ccc)-DNA were measured in tumor and nontumor tissues from the explanted livers of 16 of these 31 HCC patients and, also, in a context of tumor recurrence, in 3 patients who developed HBV/HCC recurrence. RESULTS: Fourteen patients (14.1%) developed HBV recurrence within a median period of 15 months post-OLT. HCC at OLT, a pre-OLT HBV DNA viral load > or = 100,000 copies/mL, and HBIG monoprophylaxis were independently associated with HBV recurrence post-OLT. Eleven out of the 31 patients with HCC at OLT presented with HBV recurrence and 3 out of the 68 patients without HCC had HBV recurrence (P < .0001). HBV recurrence was more frequent in patients who developed HCC recurrence (7/8 patients, 87.5%) than in those who did not (4/23 patients, 17.4%) (P < .0001). In the 16 explanted livers, cccDNA was detectable in HCC cells in 11 and in nontumor cells in 12. cccDNA was detected in a context of HCC recurrence in 2 of the 3 patients tested who developed HBV/HCC recurrence. CONCLUSIONS: The associations of HCC pre-OLT, and HCC recurrence with HBV recurrence post-OLT, and the detection of HBV DNA and cccDNA in HCC suggest that HBV replication in tumor cells may contribute to HBV recurrence post-OLT. PMID- 18424271 TI - High content analysis of gamma-secretase activity reveals variable dominance of presenilin mutations linked to familial Alzheimer's disease. AB - gamma-Secretase mediates the intramembranous proteolysis of amyloid precursor protein (APP), Notch and other cellular substrates and is considered a prime pharmacological target in the development of therapeutics for Alzheimer's disease (AD). We describe here an efficient, new, simple, sensitive and rapid assay to quantify gamma-secretase activity in living cells by flow cytometry using two membrane-bound fluorescent probes, APP-GFP or C99-GFP, as substrates for gamma secretase. The principle of the assay is based on the fact that the soluble intracellular domain of GFP-tagged APP (AICD-GFP) is released from the membrane into the cytosol following gamma-secretase cleavage. Using this feature, enzymatic activity of gamma-secretase could be deduced from the extent of the membrane retention of the probe observed after plasma membrane permeabilization and washout of the cleaved fraction. By applying two well-known gamma-secretase inhibitors (DAPT and L-685,458), we validated our assay showing that the positional GFP-based probes for gamma-secretase activity behave properly when expressed in different cell lines, providing the basis for the further development of a high-throughput and high content screening for AD targeted drug discovery. Moreover, by co-expression of different familial AD-linked mutated forms of presenilin--the key component of the gamma-secretase complex--in cells devoid of any endogenous gamma-secretase, our method allowed us to evaluate in situ the contribution of different presenilin variants to the modulation of the enzyme. PMID- 18424272 TI - Protein phosphatase 4 regulates apoptosis, proliferation and mutation rate of human cells. AB - The proteins which regulate apoptosis are of great importance both in normal cell biological processes and in the development of pathology in the diverse diseases which involve apoptosis dysfunction. The activity of many of these proteins is controlled by reversible phosphorylation, so that the relevant kinases and phosphatases play crucial roles in apoptosis control. Here we report the analysis of the role of the serine/threonine protein phosphatase, protein phosphatase 4, in controlling the apoptosis of HEK 293 T cells, using the complementary techniques of gene over-expression and down regulation through RNA interference. This analysis has demonstrated that PP4 regulates both apoptosis and proliferation in human cells and has also shown that the level of PP4 has a strong influence on gene mutation rate, which is crucial to oncogenesis. A parallel proteomic analysis has shown that the phosphorylation status of many relevant protein targets is affected by changes in PP4 and has focused attention particularly on the critical apoptosis regulators Bad and PEA-15. The phosphorylation of both of these proteins is increased when PP4 levels are suppressed, and is reduced when PP4 levels are increased, with striking consequences for the fate of the cell. PMID- 18424273 TI - Increasing carbohydrate diversity via amine oxidation: aminosugar, hydroxyaminosugar, nitrososugar, and nitrosugar biosynthesis in bacteria. AB - Bacterial secondary metabolites often contain carbohydrate attachments that play a significant role in conferring biological activity. A small proportion of these bioactive sugars are derived from aminosugar oxidation to ultimately provide hydroxyaminosugars, nitrososugars, and nitrosugars. Recent advances in the elucidation of hydroxyaminosugar-, nitrososugar-, and nitrosugar-containing natural product gene clusters have enabled the proposal of biosynthetic pathways, the in vitro characterization of aminosugar oxidases, and the structure determination of key enzymes. This article focuses upon the key enzymatic transformations in aminosugar, hydroxyaminosugar, nitrososugar, and nitrosugar biosynthesis, as well as the unique chemical reactivity of alkoxyaminosugars, with a particular focus upon developments within the past two years. PMID- 18424274 TI - Complex oligomeric structure of a truncated form of DdrA: a protein required for the extreme radiotolerance of Deinococcus. AB - In order to preserve their genome integrity, organisms have developed elaborate tactics for genome protection and repair. The Deinococcus radiodurans bacteria famous for their extraordinary tolerance toward high doses of radiations or long period of desiccation, possess some specific genes with unknown function which are related to their survival in such extreme conditions. Among them, ddrA is an orphan gene specific of Deinococcus genomes. DdrA, the product of this gene was suggested to be a component of the DNA end protection system. Here we provide a three-dimensional reconstruction of the Deinococcus deserti DdrA((1-160)) by electron microscopy. Although not functional in vivo, this truncated protein keeps its DNA binding ability at the wild-type level. DdrA((1-160)) has a complex three-dimensional structure based on a heptameric ring that can self-associate to form a larger molecular weight assembly. We suggest that the complex architecture of DdrA plays a role in the substrate specificity and favors an efficient DNA repair. PMID- 18424275 TI - The Translin/Trax RNA binding complex: clues to function in the nervous system. AB - Translin and Trax are components of an evolutionarily conserved RNA binding complex. Deletion of Translin in yeast, Drosophila and mouse produces a dramatic loss of Trax protein indicating that its stable expression is dependent on its association with Translin. Analysis of Translin KO mice has revealed multiple behavioral abnormalities and alterations in levels of transcripts encoding synaptic proteins. A confluence of localization, biochemical and RNA trafficking studies supports the view that this complex mediates dendritic trafficking of RNAs, a process thought to play a critical role in synaptic plasticity. However, further studies are needed to define its RNA cargoes, its precise role in this process, and how its binding activity and localization are regulated. Nevertheless, there is sufficient evidence to suggest that the Translin/Trax complex be included among the cadre of RNA binding complexes, such as Staufen and CPEB, that regulate dendritic trafficking of RNA in neurons. PMID- 18424276 TI - Gene regulatory networks in the early ascidian embryo. AB - Ascidians, or sea squirts, are tunicates that diverged from the vertebrate lineage early in the chordate evolution. The compact and simple organization of the ascidian genome makes this organism an ideal model system for analyzing gene regulatory networks in embryonic development. Embryos contain relatively few cells and gene activities by individual cells have been determined. Here we review and discuss advances in our understanding of the ascidian embryogenesis emerging from genomic expression studies and analyses at the single cell level. PMID- 18424277 TI - Does preoperative weight loss predict success following surgery for morbid obesity? AB - BACKGROUND: We analyzed preoperative weight loss as a predictor of postoperative success in patients after bariatric surgery. METHODS: Data were obtained from a retrospective chart review of 562 patients in a multidisciplinary obesity clinic. RESULTS: One hundred forty-six patients met the inclusion criteria (23 men and 123 women). The mean age was 39.5 years, and the mean body mass index (BMI) was 52.6 kg/m(2). Comorbid disease includes diabetes (15.7%), hypertension (30.8%), mental illness (38.4%), and musculoskeletal disease (56.8%). Procedures performed were 16 vertical band gastroplasties, 43 open gastric bypasses, 52 laparoscopic gastric bypasses, and 35 laparoscopic adjustable gastric bands. Preoperative weight change was as follows: 31 patients gained weight (21.2%), 56 patients lost weight (38.3%), and 59 patients maintained their weight (40.4%). Postoperative weight loss was not influenced by preoperative weight change among women. However, men who gained weight preoperatively had significantly worse outcomes. CONCLUSIONS: Patients may achieve satisfactory early postoperative outcomes despite inconsistent or marginal preoperative weight change. PMID- 18424278 TI - Demographic and socioeconomic trends in the use of laparoscopic appendectomy from 1997 to 2003. AB - BACKGROUND: Because there is conflicting evidence regarding the benefits of laparoscopic appendectomy, we hypothesized that there would be measurable differences in its use among various socioeconomic groups and geographic areas. METHODS: The Nationwide Inpatient Sample was queried for appendectomies performed between the years of 1997 and 2003. Rates of laparoscopic appendectomy were compared among hospital subtypes and demographic groups. RESULTS: The percentage of appendectomies performed laparoscopically has increased from 19.1% in 1997 to 37.9% in 2003. Only 11.8% of cases of complicated appendicitis were treated laparoscopically in 1997, compared with 23.5% in 2003. Nonwhite patients and those from low-income areas continue to be less likely to undergo laparoscopic appendectomy (P < .001). CONCLUSIONS: Our analysis indicates that despite expanding use of laparoscopic appendectomy nationwide, patients who live in zip codes areas with a preponderance of minorities or low-income earners are more likely to have open appendectomy. PMID- 18424279 TI - Clinical utility of type 1 growth factor receptor expression in colon cancer. AB - BACKGROUND: To evaluate the expression pattern and prognostic significance of the type 1 growth factor receptor (T1GFR) family in colon carcinoma. METHODS: Tissue microarrays were constructed using 127 tumor samples and 47 metastatic lymph nodes and T1GFR family expression was determined by immunohistochemistry. Univariate and multivariate analyses examined clinicopathologic variables for prognostic significance, and the correlation between primary and lymph node expression was determined by Spearman correlation. RESULTS: Overexpression of HER 1, HER-2, HER-3, and HER-4 in tumor samples was 32%, 1%, 12%, and 37%, respectively, and 30%, 0%, 11%, and 24% in nodal samples, respectively. On multivariate analysis, positive margins, lymphatic invasion, and HER-3 expression were significant predictors of survival outcome. There was significant correlation between tumor and regional lymph node expression for the T1GFR family members. Tumor HER-3 expression was associated with lymphatic invasion and distant recurrence. CONCLUSIONS: Tumor HER-3 expression has prognostic utility in individuals with colon carcinoma. Correlation between tumor and lymph node expression of T1GFR family members suggests that tumor receptor status may guide targeted therapy selection. PMID- 18424280 TI - Primary tumor location impacts breast cancer survival. AB - BACKGROUND: The prognostic significance of tumor location in breast cancer remains unclear. To better understand this relationship, we evaluated the Department of Defense tumor registry. METHODS: Patients with infiltrating ductal adenocarcinoma or lobular carcinoma over a 10-year period were identified and analyzed. RESULTS: Of the 13,984 tumors, 7,871 (58%) originated from the upper outer quadrant or axillary tail, whereas the remainder were found at the nipple complex (9%), upper-inner quadrant (14%), lower-inner quadrant (9%), and lower outer quadrant (10%). Univariate analysis of cancer-specific survival revealed a significant difference based on location of the primary breast cancer. Upper outer quadrant lesions were associated with an independent contribution toward a survival benefit. CONCLUSIONS: Upper-outer quadrant breast cancers have a more favorable survival advantage when compared with tumors in other locations. Factors that negatively impacted survival included high-grade tumors, advanced stage, and race. PMID- 18424281 TI - Incidence and outcomes of contralateral breast cancers. AB - BACKGROUND: The significance of a contralateral breast cancer is largely unknown, making prophylactic mastectomy controversial. METHODS: Differences between stages of initial and contralateral cancers were determined by t test. Survival distributions were compared by log-rank analyses and compared with Surveillance Epidemiology and End Results data for unilateral cancers. RESULTS: Metachronous contralateral cancers occurred at a rate of .13% per year and were of significantly lower stage. Metachronous cancers adversely impacted survival for patients with low-stage initial cancers, but the interval between cancers was less than 36 months. Synchronous tumors occurred in 2.3% of patients; survival was worse than for patients with metachronous cancers. CONCLUSIONS: Prophylactic mastectomy is unlikely to be beneficial because of the lower stages and low incidence of second cancers, even for patients with initial low-stage cancers. PMID- 18424282 TI - Inferior gluteal artery perforator flap breast reconstruction. AB - BACKGROUND: In thin patients or in those with prior surgery that precludes the use of abdominal tissue for autologous breast reconstruction, the skin and fat of the lower buttock, perfused by perforating branches of the inferior gluteal artery, has been proposed as an alternative. METHODS: This study reviewed 19 reconstructions based on the inferior gluteal artery that were performed between July 2001 and March 2007. Patient characteristics, cancer stage and treatment, indications for use of gluteal tissue, surgical time, length of hospitalization, and complications were recorded. RESULTS: Our average patient age was 49 years, with early stage breast cancer, and low body mass index. The average surgical time was 9 hours and 7 minutes, and the average hospitalization time was 4 days. Complications included 2 complete flap losses, seromas, and delayed donor site healing. CONCLUSIONS: We conclude that tissue from the lower buttock, perfused by branches of the inferior gluteal artery, is a useful alternative for autologous breast reconstruction. PMID- 18424283 TI - A remembrance of Victoria and the Canadian Army Medical Corps in the Great War. AB - The North Pacific Surgical Association first met in Victoria in December, 1917, in the midst of World War I, or as it was known then, the Great War. On all sides, the toll in human life was staggering. Canada alone lost more than 60,000 men in the war. Our Association now returns to Victoria as the very last survivors of that generation pass into history. We honor the great sacrifice of the Canadian Army, recall the horrific conditions they endured, and honor the doctors and nurses who attended the countless wounded through the experiences of a Canadian surgeon from Calgary, Dr. Harold McGill, who served for 3 years in the thick of action on the Western Front. PMID- 18424284 TI - Early "sham" feeding of neonates promotes oral feeding after delayed primary repair of major congenital esophageal anomalies. AB - BACKGROUND: To promote the appropriate and early development of the suck and swallow mechanism, we instituted a "sham" feeding protocol in infants undergoing delayed primary repair of an esophageal atresia anomaly. METHODS: Four patients who were born with esophageal atresia required delayed repair and were sham fed before surgical correction. Each infant started with a small volume of oral feeds by using a bottle/nipple with a continuous Replogle suction system placed nasally into the esophageal pouch. They were carefully monitored. The volume of feeds was slowly increased as tolerated. RESULTS: All infants successfully completed the sham feeding protocol before undergoing delayed primary esophageal repair. After repair, they had a shortened time to full oral feeding. CONCLUSION: Our "sham" feeding protocol is safe and very effective in early development of oral feeding mechanisms and shortens time to complete oral feeding after delayed esophageal repair. PMID- 18424285 TI - Left-sided gastroschisis: higher incidence of extraintestinal congenital anomalies. AB - PURPOSE: The purpose of this study was to present 3 cases of left-sided gastroschisis and review the literature concerning this rare condition. METHODS: Charts of 3 previously unreported patients with left-sided gastroschisis were reviewed. A literature review of all cases of left-sided gastroschisis was completed. RESULTS: Sixteen patients with left-sided defects were identified, only 12 of whom had classic periumbilical defects. We present 3 additional patients. Eleven of 15 patients were female, with 1 patient's sex not reported. Forty percent had extraintestinal anomalies. CONCLUSIONS: Left-sided gastroschisis is more common in females and associated with a higher incidence of extraintestinal anomalies compared with right-sided lesions. Although the etiology remains unknown, it may differ from that of right-sided gastroschisis. PMID- 18424286 TI - Evaluation of type 1 growth factor receptor family expression in benign and malignant thyroid lesions. AB - BACKGROUND: The aim of this study was to evaluate the diagnostic and prognostic utility of the type 1 growth factor receptor family in the management of differentiated thyroid cancer (DTC). METHODS: Tissue microarrays consisting of 100 benign thyroid lesions and 105 malignant thyroid lesions stained for HER1, HER2, HER3, and HER4 were evaluated. RESULTS: HER1, HER2, HER3, and HER4 were expressed in 76%, 2%, 57%, and 73% of DTC cases, respectively. HER1 and HER3 showed significantly increased expression, and HER4 showed significantly decreased expression, in DTC compared with benign thyroid lesions. HER3 expression correlated with the presence of lymph node metastasis, tumor type, and higher N stage; the expression of HER4 correlated with lower T stage. A classifier targeting benign versus malignant status with all 4 markers as potential predictors displayed an accuracy, sensitivity, and specificity of 66.8%, 63.5%, and 70.0%, respectively. CONCLUSIONS: For DTC, HER1, HER3, and HER4 have diagnostic and prognostic utility, and warrant further study as targets for cancer treatment. PMID- 18424287 TI - Pancreatic islet transplantation into the bone marrow of the rat. AB - BACKGROUND: The liver is the current site for pancreatic islet transplantation, but presents important technical complications and limitations. We asked whether pancreatic islets could be engrafted in the bone marrow, an easily accessible and widely distributed transplant site that may lack the limitations seen in the liver. METHODS: We implanted pancreatic islet isografts (Lewis islets to Lewis rats), allografts (Wistar Furth islets to Sprague Dawley rats), and xenografts (Tilapia islets to Sprague Dawley rats) into the bone marrow of nondiabetic recipients and assessed survival by histology and immunocytochemistry. No immunosuppression was used. RESULTS: Isografts and allografts showed positive staining for insulin and glucagon and no evidence of allograft rejection up to 21 days posttransplant. Xenografts were acutely rejected. CONCLUSIONS: The bone marrow may be an attractive alternative site for pancreatic islet transplantation. The acceptance of allografts and isografts but rejection of xenografts suggests a selective phenomenon for the inflammatory process. PMID- 18424288 TI - Management and outcome of pneumatosis intestinalis. AB - BACKGROUND: Pneumatosis intestinalis (PI), infiltration of gas into the bowel wall, has traditionally been associated with immediate operative intervention and a high mortality rate. METHODS: We retrospectively reviewed the diagnosis and management of pneumatosis in an attempt to characterize the disease, and examined management strategies. RESULTS: Ninety-seven patients had a computed tomography (CT) diagnosis of pneumatosis. The location of pneumatosis was as follows: 46% colon, 27% small bowel, 5% stomach, and 7% both small and large bowel. Fourteen patients also had portal venous gas and 6 (43%) of these patients died. Management strategy was non-operative in 52%, operative in 33%, and futile care in 15%. The overall mortality rate was 22% (16% operative, 6% non-operative, and 87% futile). Patients who died had a higher mean APACHE II score (25 vs 11, P <.001). CONCLUSIONS: Approximately 50% of patients with pneumatosis can be successfully managed non-operatively. The combination of PI and portal venous gas may confer a higher mortality rate. PMID- 18424289 TI - Does surgeon fatigue influence outcomes after anterior resection for rectal cancer? AB - BACKGROUND: The long work hours in surgery may contribute to medical errors and impact patient outcomes. To date, there are no studies investigating outcomes related to fatigue in general surgery. METHODS: All patients undergoing anterior resection between 1994 and 2005 at 2 university hospitals were identified. Cases were categorized as fatigued or nonfatigued and then compared with respect to complications and cancer recurrence. RESULTS: Two hundred seventy patients underwent anterior resection during the study period. Of these, 22 were performed when the surgeon was fatigued. The fatigued and nonfatigued groups had similar preoperative characteristics. The rates of intraoperative complications (fatigued 14%, rested 18%, P = .58), major postoperative complications (fatigued 9%, rested 15%, P = .68), long-term complications (fatigued 31%, rested 31%, P = .9), and local cancer recurrence rates (fatigued 0%, rested 7%, P = .2) were not significantly different between the 2 groups. CONCLUSIONS: Surgeon fatigue did not influence outcomes after anterior resection for rectal cancer. PMID- 18424290 TI - Mapping surgical practice decision making: an interview study to evaluate decisions in surgical care. AB - BACKGROUND: Training surgeons in vital decision-making skills remains unstructured. We aimed to map a process of care from the perspective of surgical decision making and test whether it could be deconstructed into an accessible format for trainees. METHODS: Semistructured interviews were conducted with 10 experienced surgeons by using symptomatic gallstones as a clinical exemplar. Data were analyzed by 2 independent reviewers to identify decisions, cues, and decision rules, with further thematic analysis of selected decisions. RESULTS: Eighteen specific decisions were identified in 6 or more transcripts, with significant interreviewer reliability (Spearman's rho = .65, P = .004, n = 17, 1 outlier excluded). These were arranged to form a "decision map." Two main decision strategies were described. CONCLUSIONS: We identified and mapped the decisions made in the care of patients with symptomatic gallstone disease. The interpretation of competency at any procedure should include the surgeon's ability to make appropriate decisions at all stages of patient care. PMID- 18424291 TI - Equine-related injury: a retrospective analysis of outcomes over a 10-year period. AB - BACKGROUND: Morbidity and financial loss caused by equine-related injuries may be significant. The purposes of this study were to determine the patterns of equine related injury and the impact on outcomes. METHODS: A 10-year retrospective review of equine-related injuries was performed. Age, gender, mechanism, injury severity score, Glasgow Coma Score, length of stay, surgical interventions, and mortality were assessed. RESULTS: Of 80 emergency department evaluations, 76 patients were admitted and form the basis of this study. The most frequent mechanism of injury was fall (68%), followed by crush injuries (15%), kicks (8%), and trampling (5%). Musculoskeletal injuries were most common (64%). Thirty-eight (50%) patients required surgical intervention. Thirty-seven (52%) patients were discharged home; 34% required outpatient physical therapy, and 14% required inpatient rehabilitation. The mortality rate was 7%. CONCLUSIONS: Equine-related injuries resulted in significant morbidity; most victims required outpatient or inpatient rehabilitation. The use of preventive strategies may minimize mortality and reduce the financial impact of postinjury morbidity. PMID- 18424292 TI - Ante-hepatic gastrojejunostomy. AB - Surgical treatment of morbid obesity continues to increase in popularity. The evolution of laparoscopic techniques has in part fueled that popularity. A common obstacle in achieving exposure of the angle of His during a laparoscopic Roux-en Y gastric bypass is a massively enlarged liver. Exposure of the angle of His is key to successfully creating the gastric pouch and completing the gastrojejunostomy. Several well-described techniques exist to achieve this exposure but suffer from potential draw backs such as bleeding and the need to insert additional trocars. We describe a technique that allows safe, excellent exposure of the gastroesophageal junction during laparoscopic Roux-en-Y gastric bypass. PMID- 18424293 TI - How to improve performance of robotic totally endoscopic coronary artery bypass grafting. AB - BACKGROUND: We investigated whether specific surgical measures during the learning curve can influence procedural performance in robotic totally endoscopic coronary artery bypass grafting (TECAB). METHODS: From 2001 to 2006, 70 patients underwent TECAB using the da Vinci telemanipulation system (Intuitive Surgical, Sunnyvale, CA). The following measures were employed with the intent to improve procedure performance: (1) introduction of a fixed team of surgeons at case 14; (2) application of fibrin glue to seal the anastomosis at case 28; and (3) use of a fourth port for transthoracic assistance at case 49. RESULTS: All 3 measures resulted in a reduction of operating room (OR) time (27%, 20%, and 20%). Use of a stable OR team and peri-anastomotic fibrin glue reduced the rate of conversions and on-table revisions from 39% to 9% (P = .006) and from 26% to 7% (P = .038), respectively. Peri-anastomotic fibrin glue significantly reduced postoperative revisions for bleeding. Using transthoracic assistance, anastomotic time was shortened from 35 minutes (range 23-67) to 25 minutes (range 16-100) (P < .001). CONCLUSION: The 3 measures can improve intraoperative and clinical results in TECAB. PMID- 18424294 TI - Re: Polyethylene glycol versus sodium phosphate mechanic bowel preparation in elective colorectal surgery. PMID- 18424295 TI - Re: Implications of negative technetium 99m-labeled red blood cell scintigraphy in patients presenting with lower gastrointestinal bleeding. PMID- 18424296 TI - An important study comparing an endoscopic and a medical approach in treating acute intracerebral hematomas. Editorial. PMID- 18424297 TI - Research news and notes. PMID- 18424298 TI - Image-guided endoscopic evacuation of spontaneous intracerebral hemorrhage. AB - BACKGROUND: Spontaneous ICH is a devastating disease with high morbidity and mortality. Intracerebral hemorrhage lacks an effective medical or surgical treatment despite the acknowledged pathophysiologic benefits of achieved hemostasis and clot removal. Image-guided stereotactic endoscopic hematoma evacuation is a promising minimally invasive approach designed to limit operative injury and maximize hematoma removal. METHODS: A single-center randomized controlled trial was designed to assess the safety and efficacy of stereotactic hematoma evacuation compared to best medical management. Patients were randomized within 24 hours of hemorrhage in a 3:2 fashion to best medical management plus endoscopic hematoma evacuation or best medical management alone. Data were collected to assess efficacy and safety of hematoma evacuation and to identify procedural components requiring technical improvement. RESULTS: Ten patients have been enrolled and randomized to treatment. Six patients underwent endoscopic evacuation with a hematoma volume reduction of 80% +/- 13% at 24 hours post procedure. The medical arm demonstrated a hematoma enlargement of 78% +/- 142% during this same period. Rehemorrhage rates and deterioration rates were similar in the 2 groups. Mortality was 20% in the endoscopic group and 50% in the medical treatment cohort. The endoscopic technique was shown to be effective in identification and evacuation of hematomas, whereas reduction in the number of endoscopic passes and maintenance of hemostasis require further study. CONCLUSION: Image-guided stereotactic endoscopic hematoma removal is a promising minimally invasive technique that is effective in immediate hematoma evacuation. This technique deserves further investigation to determine its role in ICH management. PMID- 18424300 TI - Invasive intracranial aspergillosis: the management dilemmas. AB - BACKGROUND: Invasive intracranial aspergillosis remains a disease with high morbidity and mortality. The rapid increase in the incidence of this disease led us to review the literature and formulate a treatment protocol for such patients. METHODS: An analysis of 46 patients with invasive intracranial aspergillosis is presented and the subgroups of extradural and intradural variety are evaluated with different treatment strategies. RESULTS: Patients with extradural form of disease had 100% survival, whereas antifungal chemotherapy preloading tends to provide a better outcome. CONCLUSIONS: Extradural aspergillosis does not need chemotherapy preloading, whereas intradural variant may have improved survival chances after preloading. Liposomal formulation has an advantage of shortened time duration for preloading, because the daily administration dose is 6 times higher than conventional preparation. PMID- 18424302 TI - Profiles in volunteerism Gary Heit, MD, PhD, and Americare Neurosurgery International. PMID- 18424303 TI - Therapeutic nihilism and the elderly. PMID- 18424304 TI - The health of the health service's staff in the UK. PMID- 18424305 TI - Good deeds and Google. PMID- 18424306 TI - Stopping trials early for benefit: too good to be true. PMID- 18424307 TI - The search for effective and safe disease control in psoriasis. PMID- 18424308 TI - New diagnostic strategies for pulmonary embolism. PMID- 18424309 TI - Approaching an age of reason with antiplatelet therapy. PMID- 18424310 TI - Antibiotic prophylaxis for infective endocarditis. PMID- 18424311 TI - Public-health crisis after the election violence in Kenya. PMID- 18424312 TI - Do classical origins of medical terms endanger patients? PMID- 18424314 TI - The body. PMID- 18424313 TI - Martina Potschke-Langer. PMID- 18424315 TI - Ambulatory oral amoxicillin for severe pneumonia in children. PMID- 18424316 TI - Ambulatory oral amoxicillin for severe pneumonia in children. PMID- 18424317 TI - Ambulatory oral amoxicillin for severe pneumonia in children. PMID- 18424318 TI - Ambulatory oral amoxicillin for severe pneumonia in children. PMID- 18424321 TI - Folic acid fortification and cancer risk. PMID- 18424322 TI - Folic acid fortification and cancer risk. PMID- 18424323 TI - Efficacy of ISA247 in plaque psoriasis: a randomised, multicentre, double-blind, placebo-controlled phase III study. AB - BACKGROUND: The use of systemic calcineurin inhibitors for the treatment of patients with psoriasis is limited by toxicity, particularly nephrotoxicity. ISA247, a novel inhibitor, was effective and well tolerated in a phase II study of patients with plaque psoriasis. Therefore its efficacy was assessed in this phase III study. METHODS: 451 patients aged 18-65 years with plaque psoriasis involving at least 10% of the body surface area were randomly assigned in equal proportions to receive placebo or ISA247 at 0.2 mg/kg, 0.3 mg/kg, or 0.4 mg/kg orally twice a day in dermatology clinics. The primary endpoint was a 75% reduction in the psoriasis area and severity index (PASI 75) score at week 12. Treatment allocation was concealed from patient and physicians doing the assessments by use of sealed envelopes. The method of analysis was by modified intention to treat. The trial is registered at ClinicalTrials.gov, number NCT00244842. FINDINGS: 107, 113, and 116 patients were assigned to the ISA247 0.2 mg/kg, 0.3 mg/kg, and 0.4 mg/kg groups, respectively, and 115 to the placebo group. At week 12, PASI 75 scores were achieved in the ISA247 0.2 mg/kg, 0.3 mg/kg, and 0.4 mg/kg groups by 14 (16%; 95% CI 9-24) of 105, 26 (25%; 17-24) of 111, and 44 (47%; 27-57) of 113 patients, respectively, and in the placebo group by 4 (4%; 0-8) of 113 patients. Efficacy was maintained during 24 weeks. Mild to moderate glomerular filtration rate reductions were noted in seven patients in the ISA247 0.4 mg/kg group and in one in the ISA247 0.3 mg/kg group. ISA247 blood concentrations showed a strong correlation with mean percentage reduction in PASI. INTERPRETATION: ISA247 was safe and effective in the treatment of patients with moderate to severe psoriasis during 24 weeks, with the highest dose providing the best efficacy. The strong correlation between ISA247 concentrations and efficacy might allow for accurate dosing of patients compared with existing calcineurin inhibitors. PMID- 18424325 TI - Prevalence, pathogenesis, and causes of chronic cough. AB - Cough is a reflex action of the respiratory tract that is used to clear the upper airways. Chronic cough lasting for more than 8 weeks is common in the community. The causes include cigarette smoking, exposure to cigarette smoke, and exposure to environmental pollution, especially particulates. Diseases causing chronic cough include asthma, eosinophilic bronchitis, gastro-oesophageal reflux disease, postnasal drip syndrome or rhinosinusitis, chronic obstructive pulmonary disease, pulmonary fibrosis, and bronchiectasis. Doctors should always work towards a clear diagnosis, considering common and rare illnesses. In some patients, no cause is identified, leading to the diagnosis of idiopathic cough. Chronic cough is often associated with an increased response to tussive agents such as capsaicin. Plastic changes in intrinsic and synaptic excitability in the brainstem, spine, or airway nerves can enhance the cough reflex, and can persist in the absence of the initiating cough event. Structural and inflammatory airway mucosal changes in non-asthmatic chronic cough could represent the cause or the traumatic response to repetitive coughing. Effective control of cough requires not only controlling the disease causing the cough but also desensitisation of cough pathways. PMID- 18424324 TI - Diagnosis of pulmonary embolism by multidetector CT alone or combined with venous ultrasonography of the leg: a randomised non-inferiority trial. AB - BACKGROUND: Multislice CT (MSCT) combined with D-dimer measurement can safely exclude pulmonary embolism in patients with a low or intermediate clinical probability of this disease. We compared this combination with a strategy in which both a negative venous ultrasonography of the leg and MSCT were needed to exclude pulmonary embolism. METHODS: We included 1819 consecutive outpatients with clinically suspected pulmonary embolism in a multicentre non-inferiority randomised controlled trial comparing two strategies: clinical probability assessment and either D-dimer measurement and MSCT (DD-CT strategy [n=903]) or D dimer measurement, venous compression ultrasonography of the leg, and MSCT (DD-US CT strategy [n=916]). Randomisation was by computer-generated blocks with stratification according to centre. Patients with a high clinical probability according to the revised Geneva score and a negative work-up for pulmonary embolism were further investigated in both groups. The primary outcome was the 3 month thromboembolic risk in patients who were left untreated on the basis of the exclusion of pulmonary embolism by diagnostic strategy. Clinicians assessing outcome were blinded to group assignment. Analysis was per protocol. This study is registered with ClinicalTrials.gov, number NCT00117169. FINDINGS: The prevalence of pulmonary embolism was 20.6% in both groups (189 cases in DD-US-CT group and 186 in DD-CT group). We analysed 855 patients in the DD-US-CT group and 838 in the DD-CT group per protocol. The 3-month thromboembolic risk was 0.3% (95% CI 0.1-1.1) in the DD-US-CT group and 0.3% (0.1-1.2) in the DD-CT group (difference 0.0% [-0.9 to 0.8]). In the DD-US-CT group, ultrasonography showed a deep-venous thrombosis in 53 (9% [7-12]) of 574 patients, and thus MSCT was not undertaken. INTERPRETATION: The strategy combining D-dimer and MSCT is as safe as the strategy using D-dimer followed by venous compression ultrasonography of the leg and MSCT for exclusion of pulmonary embolism. An ultrasound could be of use in patients with a contraindication to CT. PMID- 18424326 TI - Management of chronic cough. AB - Cough that remains unexplained after basic clinical assessment is a common reason for referral to secondary care. Much of the evidence about management of isolated chronic cough is derived from case series; this evidence suggests that isolated chronic cough is usually due to asthma, gastro-oesophageal reflux disease, and upper airway conditions, and that it can be cured in most people by treatment of these conditions. However, there is increasing recognition that satisfactory control of chronic cough is not achieved in a substantial number of patients seen in secondary care. Moreover, there is a concern that perpetuation of the belief that chronic cough is solely due to the effects of comorbid conditions is inhibiting research into the pathophysiology of an abnormally heightened cough reflex, and jeopardising development of improved treatments. We advocate a change in emphasis, which makes a clear distinction between cough due to corticosteroid responsive eosinophilic airway diseases and corticosteroid-resistant non eosinophilic cough. We recommend that some factors with weak evidence of an association with cough are best viewed as potential aggravating factors of an intrinsic abnormality of the cough reflex, rather than the cause. We call for more research into the basic mechanisms and pharmacological control of an abnormally heightened cough reflex, and recommend ways to assess the effects of potentially antitussive treatments. PMID- 18424327 TI - Immunisation against poliomyelitis: moving forward. PMID- 18424328 TI - Another reason to dislike medication. PMID- 18424329 TI - Foreword immunodeficiency--improving the deficiency of knowledge. PMID- 18424330 TI - Immunodeficiency disorders. Preface. PMID- 18424331 TI - From infectious diseases to primary immunodeficiencies. AB - The field of primary immunodeficiencies has expanded, thanks to the exploration of novel clinical phenotypes and their connection with morbid genotypes, and the subsequent exploration of new patients who have known primary immunodeficiency defining clinical phenotypes and their connection with novel morbid genotypes. This two-way process is becoming increasingly active, particularly for patients who have infectious diseases in whom the underlying immunologic and genetic causes remain mostly unexplained. The authors review how the exploration of children who have clinical infectious diseases caused by mycobacteria, pneumococcus, and herpes simplex virus recently led to the description of three new groups of primary immunodeficiencies. These three examples justify the continuation of the genetic exploration of novel infectious phenotypes and novel patients who have infections. This challenging process will eventually reap its rewards, to the benefit of patients and their families. PMID- 18424332 TI - Congenital neutropenia syndromes. AB - Congenital neutropenia syndromes comprise a heterogeneous group of inherited disorders. Hereditary conditions associated with low neutrophil counts are persistent and need to be differentiated from neutropenia secondary to autoimmune processes or other pathologic conditions, such as myelodysplasia or leukemia. Clinically, congenital neutropenia is characterized by recurrent bacterial infections. Recently, several novel genetic defects were described in patients with congenital neutropenia, shedding light on the pathophysiology of these rare diseases. PMID- 18424333 TI - The hyper-IgE syndromes. AB - The hyper IgE syndromes (HIES) are rare primary immune deficiencies characterized by elevated serum IgE, rash, and recurrent bacterial infections of the skin and lung. Autosomal dominant HIES, the most common disease in this group, results from STAT3 mutations and has a variety of connective tissue and skeletal abnormalities. The genetic etiologies of the more rare autosomal recessive forms still need delineation. Treatment of these syndromes has relied on prophylactic and therapeutic antimicrobial agents and aggressive skin care. The new and evolving genetic and immunologic understandings of this previously elusive set of diseases should lead to more effective disease-specific therapies. PMID- 18424334 TI - Hemophagocytic lymphohistiocytosis and other hemophagocytic disorders. AB - Hemophagocytic disorders result when critical regulatory pathways responsible for the natural termination of immune/inflammatory responses are disrupted or overwhelmed. Hemophagocytic disorders reflect pathologic defects that alter the normal crosstalk between innate and adaptive immune responses, and compromise homeostatic removal of cells that are superfluous or dangerous to the organism. Although hemophagocytic disorders are considered rare, increased awareness of these conditions has led to more frequent diagnoses, more rapid initiation of life-saving treatments, and new insights into the molecules and pathways involved in natural immune down-regulation. Furthermore, improved understanding of the immunologic abnormalities revealed by hemophagocytic disorders informs potential new treatments for life-threatening multisystem organ dysfunction related to sepsis in the intensive care unit setting and severe cases. PMID- 18424335 TI - Immune dysregulation in primary immunodeficiency disorders. AB - The past several years have brought an increased awareness of the prevalence of autoimmunity and immune dysregulation among patients who have primary immunodeficiency disorders (PIDD). The recent clinical and molecular definition of PIDD, in which the primary defect is in the immunoregulatory compartment of the immune system, has offered insight into the basic mechanisms of immune tolerance, which has provided new targets and new techniques to study immune tolerance in PIDD. Many of these studies have focused on the presence and function of regulatory T (T(REG)) cells in PIDD, particularly since the discovery of murine and human syndromes associated with T(REG) deficiency. This article focuses on the current state of knowledge regarding the role of T(REG) in various PIDD that have clinical features indicative of dysregulated immunity. PMID- 18424337 TI - Chromosome 22q11.2 deletion syndrome: DiGeorge syndrome/velocardiofacial Syndrome. AB - DiGeorge syndrome, or chromosome 22q11.2 deletion syndrome, is a disorder affecting multiple organ systems. The immunologist may be called on to coordinate complex medical care tailored to the specific needs and unique clinical features of each patient. This article focuses on the immune system, but patients require a holistic approach. Attention to cardiac, nutritional, and developmental needs in early infancy is important, and it is critical to identify the rare infants who require either a lymphocyte or thymus transplant. Later, speech and school issues dominate the picture. Allergies and autoimmune disorders also may be troubling for some school-age children. PMID- 18424336 TI - Genetic defects of apoptosis and primary immunodeficiency. AB - Programmed cell death is important for maintaining lymphocyte homeostasis. Several human-inherited diseases with impaired apoptosis have been identified at the genetic level: autoimmune lymphoproliferative syndrome, caspase-8 deficiency state, and X-linked lymphoproliferative syndrome. These diseases feature excess lymphocyte accumulation, autoimmunity, or immunodeficiency. Elucidating their molecular pathogenesis has also provided new insights into the signaling mechanisms regulating apoptosis and lymphocyte activation. PMID- 18424338 TI - Common variable immunodeficiency: an update on etiology and management. AB - Common variable immunodeficiency (CVID) represents a heterogeneous group of primary antibody deficiency disorders characterized by recurrent infection and by inflammatory, granulomatous, and autoimmune complications. Recently, there have been significant advances in understanding the pathogenesis of the disease, with five genetic mutations identified in patients who have a CVID phenotype. Clinical care also has progressed with refinements in treatment and the development of classification schemes for prognostic and research purposes. Significant delays in diagnosis remain, however. It is likely that more genetic defects will be identified in the future, further shrinking the pool of patients who have CVID of unknown cause. PMID- 18424339 TI - Genetic diagnosis of primary immune deficiencies. AB - Gene testing in primary immune deficiencies (PIDs) once was limited to expert academic laboratories, but now is easily available to physicians with a broad range of clinical expertise. Such testing can establish or confirm a suspected diagnosis and also may predict future disease risk in advance of clinical signs and symptoms, inform reproductive decision making, and guide clinicians in selecting the most appropriate therapeutic options. This article, based on the authors' experience and a review of the published literature, discusses some of the advances and challenges currently encountered in the clinical molecular genetic diagnosis of PIDs. PMID- 18424340 TI - Principles of and advances in immunoglobulin replacement therapy for primary immunodeficiency. AB - During the last 2 decades, the continued development and the large-scale production of polyclonal immune serum globulin (ISG) preparations with improved safety and tolerability profiles have allowed treatment to focus on quality of life and long-term freedom from the complications of primary immune deficiency disease, rather than just on freedom from severe acute infections and survival. Available ISG preparations allow routine therapy by a variety of routes and regimens that can be tailored to suit individual patients. Continued vigilance is required, however, because problems with emerging diseases, and the costs and availability of ISG are likely to present continuing challenges. PMID- 18424341 TI - Advances in hematopoietic stem cell transplantation for primary immunodeficiency. AB - The molecular bases of most primary immunodeficiencies (PID) have been discovered. Long-term follow-up of patient cohorts treated with antimicrobial prophylaxis has demonstrated good short-term prognosis but with increasing morbidity and mortality over time. The results of hematopoietic stem cell transplantation (HSCT) for PID have improved incrementally over time, with survival and cure of 90% for some defined diseases. This article examines the advances in HSCT for PID and argues that HSCT should be considered earlier for most patients. PMID- 18424342 TI - Gene therapy for primary immunodeficiencies. AB - Primary immunodeficiencies are a group of disorders that are highly amenable to gene therapy because of their defined pathophysiology and the accessibility of the hematopoietic system to molecular intervention. The development of this new therapeutic modality has been driven by the established morbidity and mortality associated with conventional allogeneic stem cell transplantation, particularly in the human leukocyte antigen-mismatched setting. Recently, several clinical studies have shown that gamma retroviral gene transfer technology can produce major beneficial therapeutic effects, but, as for all cellular and pharmacologic treatment approaches, with a finite potential for toxicity. Newer developments in vector design showing promise in overcoming these issues are likely to establish gene therapy as an efficacious strategy for many forms of primary immunodeficiencies. PMID- 18424343 TI - Genetics in critical care. PMID- 18424344 TI - Genetics in critical care: a toolbox. AB - Becoming comfortable with the onslaught of genetic information requires a familiarity with genetics terminology and basic molecular genetics. Caring for the genetic health of our patients requires an additional appreciation of genetic testing and screening issues and new areas of study with strange-sounding names, such as proteomics, nutrigenomics, and epigenetics. This article provides an overview of basic genetic principles and terminology with applications to critical care patients. It lays a foundation to support understanding of the other articles in this issue. An appreciation of genetics (the study of heredity) and genomics (the study of the interaction of genes within an organism) has an important place in the knowledge base of every critical care clinician. PMID- 18424345 TI - The family history: reemergence of an established tool. AB - Genetics has transformed the use of family history information and has led to the reemergence of the detailed genetic family history. It is critical that public and professional educational efforts to increase family history awareness and working knowledge are prioritized. Patient maintenance of the pedigree provides increased patient awareness and facilitates some of the limitations associated with conventional medical history ascertainment, ultimately improving health care and research. The increasing use of genetic screening promises to cultivate a paradigm shift in medical treatment emphasizing primary prevention and early intervention. Appreciation of the family history is necessary to make this important advance. PMID- 18424346 TI - Congenital heart disease and genetics. AB - Improvements in the diagnosis and treatment of congenital heart disease have drastically reduced the morbidity and mortality associated with such defects. Knowledge regarding the genetic contributions to congenital heart disease is considered to be in its infancy; however, the field of cardiovascular genetics in humans is moving at a rapid pace. This article discusses what is currently known about the genetic contribution to congenital heart disease, including structural defects and congenital cardiac arrhythmias. Genetic and chromosomal syndromes that involve the heart are reviewed along with genetic testing. PMID- 18424347 TI - Genetic influences in common respiratory disorders. AB - Respiratory disorders are common problems for adults and children in North America and generally represent the outcome of gene-environment interactions. Some problems are considered genetic in origin, such as cystic fibrosis, and others are considered environmental in origin, such as respiratory infections. Emerging information indicates that even genetic-based disorders are influenced by the environment and that environmental-based disorders are modified by personal genetic factors in individual physiologic responses. An understanding of an individual's personal risk factors for disease or health problem development can allow health care professionals to tailor health promotion strategies and treatment plans with appropriate environmental manipulation. This article explores the genetic influences that may affect the individual's physiologic responses and the consequences of environmental stimuli. PMID- 18424348 TI - Hereditary hemochromatosis: pathophysiology, diagnosis, and management. AB - Hereditary hemochromatosis (HH) is an autosomal recessive genetic disease resulting in inappropriate intestinal iron absorption leading to iron overload and end-organ disease. The disease is most prevalent in white individuals of European descent. The C282Y mutation on the HFE gene accounts for most cases of HH; however, other genetic mutations have been identified. End-organ damage results in cirrhosis, diabetes mellitus, and cardiomyopathy. Therapeutic phlebotomy to deplete excessive iron stores is the standard treatment of HH and results in normal longevity if therapy is initiated before end-organ disease occurs. PMID- 18424349 TI - Genes and acute neurologic disease and injury: a primer for the neurologic intensive care nurse. AB - The genetics revolution has not evaded the neuroscience community. Neurologic diseases and injuries, particularly of the type seen in neurologic intensive care units, are difficult to treat and often have poor prognoses. Recent work has begun to identify genotype-specific influences on development and treatment of multiple sclerosis and stroke. Additionally, responses to diseases and injuries to the brain and spinal cord have genetic influences. This article informs nurses working with neurologically impaired patients in the intensive care unit of specific genes involved in patient response and potential future therapeutics. PMID- 18424350 TI - Inflammation and genomics in the critical care unit. AB - Inflammation is a physiologic response to irritants, injury, and infection. Inflammatory dysregulation is believed to contribute to mortality and morbidity in illnesses common to critical care units, such as burns, trauma, sepsis, and cardiovascular disease. This article reviews the approaches used to investigate the molecular basis of inflammatory function. Genomic findings are providing insight into clinical diagnosis and treatment of inflammatory derangements in critically ill patients. PMID- 18424351 TI - Pharmacogenetics in critical care: atrial fibrillation as an exemplar. AB - Pharmacogenetic testing is currently not routine in critical care settings but recent changes in the warfarin label are likely to lead to critical care nurses encountering physician or nurse practitioner orders for such testing. Although the science for pharmacogenetics is complex, the components of patient teaching are not beyond that which nurses already provide about other laboratory, disease, and treatment-based information. It is reasonable to expect that as the science of pharmacogenetics and pharmacogenomics expands and discoveries are translated in clinical settings, the additional information from pharmacogenetic test results will help prescribers select or adjust medication doses to reduce the risk for adverse drug reactions and improve the chances of achieving therapeutic targets in a timely fashion. PMID- 18424353 TI - The -1131T>C polymorphism in the apolipoprotein A5 gene is related to hypertriglyceridemia in Taiwanese aborigines. AB - The prevalence of hypertriglyceridemia, considered to be an independent risk factor for the development of cardiovascular disease, is high in Taiwanese aborigines. This study was undertaken to examine the effect of the -1131T>C polymorphism in the apolipoprotein A5 gene on serum triglyceride levels in female Taiwanese aborigines. This was a cross-sectional study, and a total of 316 unrelated female Taiwanese aborigines were genotyped at the -1131T>C polymorphism in apolipoprotein A5 using the polymerase chain reaction-restriction fragment length polymorphism method. Serum triglyceride > or = 150 mg/dL was defined as the hypertriglyceridemia group and triglyceride < 150 mg/dL was considered to be the control group. The frequency of the minor C allele was significantly higher in the hypertriglyceridemia group (0.53) than in the control group (0.35) (p < 0.001). The frequency of this rare allele was comparable to that in Japanese and Han Chinese, but was higher than that in Caucasians. In a multiple logistic model adjusted for possible confounders, C allele-containing variants were independently associated with greater risks (CT genotype: OR = 3.28, 95% CI = 1.43-7.56; CC genotype: OR = 5.86, 95% CI = 2.15-15.99) of hypertriglyceridemia than the TT genotype (p < 0.01), notably with the CC homozygote exhibiting the greatest risks. The genotype polymorphisms were also associated with serum triglyceride concentrations in a linear fashion (for trend, p < 0.05). Our results indicate that the -1131T>C polymorphism of the Apo A5 gene influences serum triglyceride levels in female Taiwanese aborigines, and that differences exist in the frequency of the C allele among people of various ethnicities. PMID- 18424352 TI - Neonatal genetic testing is more than screening. AB - Newborn screening practices have changed since breakthroughs have occurred in genetics and mapping of the human genome. Although newborn screening has been in existence since the 1960s, today's newborn screening practices are subsumed primarily under the umbrella of genetic testing. Inclusion of the family history tool is another dimension of neonatal assessment. Technology allows many noninvasive tests to be run at a low cost but with this advance comes ethical and legal dilemmas. This article discusses neonatal genetic testing and some of the ethical dilemmas that arise. PMID- 18424354 TI - Low-energy helium-neon laser therapy induces repigmentation and improves the abnormalities of cutaneous microcirculation in segmental-type vitiligo lesions. AB - Segmental vitiligo (SV) is a special form of vitiligo occurring in a dermatomal distribution, and an abnormality involving the sympathetic nerves supplying the affected dermatome is known to underlie this disorder. Previously, we have shown that SV is associated with an abnormal increase in cutaneous blood flow and adrenoceptor responses in the affected areas. Since SV is resistant to conventional forms of therapy, its management represents a challenge for dermatologists. Low energy helium-neon lasers (He-Ne laser, wavelength 632.8 nm) have been employed as a therapeutic instrument in many clinical situations, including vitiligo management and repair of nerve injury. The purpose of this study was to evaluate the effectiveness and safety of He-Ne lasers in treating SV, and determine their effects on the repair of sympathetic nerve dysfunction. Forty patients with stable-stage SV on the head and/or neck were enrolled in this study. He-Ne laser irradiation was administered locally at 3.0 J/cm2 with point stimulation once or twice weekly. Cutaneous microcirculatory assessments in six SV patients were performed using a laser Doppler flowmeter. The sympathetic adrenoceptor response of cutaneous microcirculation was determined by measuring cutaneous blood flow before, during and after iontophoresis with sympathomimetic drugs (phenylephrine, clonidine and propranolol). All measurements of microcirculation obtained at SV lesions were simultaneously compared with contralateral normal skin, both before and after He-Ne laser treatment. After an average of 17 treatment sessions, initial repigmentation was noticed in the majority of patients. Marked repigmentation (> 50%) was observed in 60% of patients with successive treatments. Cutaneous blood flow was significantly higher at SV lesions compared with contralateral skin, but this was normalized after He-Ne laser treatment. In addition, the abnormal decrease in cutaneous blood flow in response to clonidine was improved by He-Ne laser therapy. Our study showed that He-Ne laser therapy is an effective treatment for SV by normalizing dysfunctions of cutaneous blood flow and adrenoceptor responses in SV patients. Thus, the beneficial effects of He-Ne laser therapy may be mediated in part by a reparative effect on sympathetic nerve dysfunction. PMID- 18424355 TI - Potential risk and protective factors for in-hospital mortality in hyperacute ischemic stroke patients. AB - In the era of thrombolytic therapy for hyperacute ischemic stroke, most investigators have focused their attention on the factors influencing mortality and functional outcomes in patients treated with thrombolysis, but very few have focused on these factors among patients not receiving thrombolysis. The aim of this study was to investigate the prognostic factors for mortality in all hyperacute stroke patients with or without thrombolysis. In 2005, we enrolled 101 ischemic stroke patients (43 females, 58 males; mean age, 68 years) who were transported to the emergency department (ED) within 4 hours of symptom onset. The overall in-hospital mortality rate was 17.8% (18/101). According to t test analysis, age (p = 0.034), time interval from neurologist consultation (p < 0.0001) and ED to ward admission (p = 0.001), Glasgow coma scale (GCS) (p = 0.001), National Institutes of Health Stroke Scale (NIHSS) (p < 0.0001) and the sum of major risk factors of cerebrovascular disease (CVD) (p < 0.0001) were significantly different between mortality and survivor groups. Further Chi squared test analysis revealed significant differences in the presenting consciousness disturbance (p = 0.001), place of attack (p = 0.04), and referral transportation (p = 0.008) between these groups. In conclusion, old age, delay between neurologist consultation and ward admission, severity of stroke, and multiple risk factors of CVD are significant risk factors for in-hospital mortality. Conversely, being free of initial consciousness disturbance, living in an urban area, and having direct transportation to a stroke center are protective factors in survivors. The concept of "brain attack" should be re-emphasized among ED physicians. The interconnection between stroke centers and emergency medical systems (EMS) should be more tightly built to promote timely management for hyperacute stroke care. PMID- 18424356 TI - Cognitive and motor components of response speed in the stroop test in Parkinson's disease patients. AB - This study assessed the performance of patients with Parkinson's disease (PD) in the Stroop test. Twenty-seven patients with PD (17 men, 10 women; mean age, 63.3 +/- 10.5 years) and 27 age-matched controls (14 men, 13 women; mean age, 63.5 +/- 9.2 years) were administered the color-naming, word-reading, and incongruent color-word-naming tasks in the Stroop test. Compared with the normal control group, the PD group had slower speeds for all three tasks and greater Stroop interference, indicating a response inhibition deficit in PD patients. Further analysis indicated that slowness during color naming might be due to motor slowness, rather than a central cognitive processing problem in color discrimination. In conclusion, the performance of the PD group on the three tasks of the Stroop test suggests that the PD patients were deficient in motor responses and cognitive inhibitory abilities. PMID- 18424357 TI - Comparison of percutaneous nephrolithotomy and ureteroscopic lithotripsy in the management of impacted, large, proximal ureteral stones. AB - The optimal treatment for large, impacted, proximal ureteral stones remains controversial. We report our experience and compare treatment outcomes in patients with single, impacted, proximal ureteral stones undergoing percutaneous nephrolithotomy (PCNL) and ureteroscopic lithotripsy (URSL). Between January 2005 and January 2006, a total of 53 consecutive patients with solitary, impacted, proximal ureteral stones > 15 mm in diameter who had undergone PCNL or URSL treatments were enrolled in this study. The mean age was 48.5 +/- 11.8 years. PCNL and URSL were performed in 22 and 31 patients. Stone burdens in the PCNL and URSL groups were 232.8 +/- 113.2 mm2 and 150.3 +/- 70.3 mm2, respectively. The efficiency quotient (EQ) for the PCNL and URSL groups was 0.95 and 0.67, respectively. The stone-free rate at the 1 month follow-up was 95.4% in the PCNL group and 58% in the URSL group (p < 0.001). Two patients in the PCNL group had blood loss requiring transfusion. Eight patients had stones showing upward migration during the URSL procedure, and these stones were subsequently treated by extracorporeal shock wave lithotripsy and PCNL. For an impacted, proximal ureteral stone > 15 mm in diameter, PCNL had better stone-free rates and could simultaneously treat coexisting renal stones. However, URSL had the advantages of shorter operative times, shorter postoperative hospital stays, and fewer postoperative complications. PMID- 18424358 TI - Duplicated vas deferens: a rare congenital abnormality. AB - Duplication of vas deferens is a rare congenital anomaly for which the overall incidence in the general population is estimated to be less than 0.05%. We report here a case of duplicated vas deferens found during a routine varicocele operation. PMID- 18424359 TI - Giant invasive schwannoma of cauda equina with minimal neurologic deficit: a case report and literature review. AB - A 53-year-old man presented with a history of slight weakness in the right lower limb. Giant invasive cauda equina schwannoma was diagnosed according to the criteria of Sridhar et al. Schwannomas are usually benign and common tumors arising from nerve sheath cells, particularly from sensory nerves. Giant invasive schwannomas, however, are rare, and most of patients with them present with severe neurologic deficits independent of daily activity, although in the case presented here, in spite of the large size of the tumor causing pedicle erosion, expansive destruction of the vertebral body and widening of the neural foramina, there were only minimal neurologic deficits. We have therefore decided to report this case, with a review of the relevant English literature emphasizing clinical presentations, plain film images and magnetic resonance image findings of giant invasive cauda equina schwannoma for early diagnosis and differential diagnosis. PMID- 18424360 TI - Tumor-like cerebral perivasculitis in a pediatric patient with systemic lupus erythematosus. AB - Nervous system manifestations are present in up to 70% of patients with systemic lupus erythematosus (SLE). The spectrum of clinical symptoms varies widely, from severe, life-threatening symptoms at presentation, such as transverse myelitis, to symptoms of more subtle and subclinical abnormalities of neurocognitive function. We report the case of a 14-year-old patient with SLE and lupus nephritis under regular steroid therapy, who had a sudden onset of consciousness change. Brain magnetic resonance imaging showed a huge mass lesion. After surgical decompression and corticosteroid pulse therapy, the patient's neurologic symptoms improved dramatically. Brain biopsy revealed perivasculitis of the brain with marked perivascular infiltration of eosinophils, macrophages, and neutrophils. Microhemorrhage was also evident. The patient recovered without obvious neurologic sequelae. PMID- 18424361 TI - Activated protein C ratio, plasma tissue factor activity and activated factor VII in Chinese patients with coronary heart disease. AB - OBJECTIVE: There is close relationship between abnormal coagulation system and progression of coronary heart disease (CHD), our purpose is to evaluate the contribution of hematologic factors and some other risk factors to the development of coronary heart disease (CHD) in Chinese population. METHODS: 56 patients with CHD at admission and 54 controls were enrolled. Plasma levels of protein C, free protein S, total protein S, thrombomodulin, activated factor VII (FVIIa), factor VII:Ag, P-selectin, tissue-type plasminogen activator, plasminogen activator inhibitor-1 were measured by enzyme linked immunosorbent assay, activity of tissue factor (aTF) by chromogenic activity assay, and activated protein C (APC) ratio, prothrombin time, aPTT, fibrinogen, D-dimmer and thrombin time by full-automated coagulation analyzer. RESULTS: Compared with controls, plasma level of thrombomodulin, FVIIa, factor VII:Ag and aTF were raised in CHD group (p<0.05, 0.001, 0.05, and 0.05, respectively). The average APC ratio in CHD group was lower than that in controls (p<0.001). The result of binary logistic regression analysis showed that activated factor VII (OR2.680, 95%CI1.539-4.665) and tissue factor activity (OR1.019, 95%CI1.004-1.035) were risk factors and high density lipoprotein (OR0.008, 95%CI0-0.478) and activated protein C ratio (OR0.001, 95%CI0-0.011) were protective factors for CHD. CONCLUSIONS: Low activated protein C ratio, elevated tissue factor activity and increased activated factor VII in plasma may contribute to development of coronary heart disease. PMID- 18424362 TI - High performance liquid chromatographic method for the determination of HIV-1 protease inhibitor tipranavir in plasma of patients during highly active antiretroviral therapy. AB - A new high-performance liquid chromatographic method for the determination of tipranavir in human plasma is described. Quantitative recovery following liquid liquid-extraction with diethylether from 100 microl of human plasma was achieved. Subsequently, the assay was performed with 67 mM potassium dihydrogen phosphate acetonitrile as a mobile phase, a Phenomenex C 18 column and UV detection at 255 nm. Linear Standard curves were obtained for concentrations ranging from 2.5 to 400 microg/ml. The calculated intra- and inter-day coefficents of variation were below 7%. PMID- 18424363 TI - HIV-1 infected Germans have more variations on neck region of DC-specific intercellular adhesion molecule-3-grabbing nonintegrin than HIV-1 infected Chinese. AB - The C-type lectins DC-SIGN and DC-SIGNR bind and transmit HIV-1 depending on the tetramer formed by "neck" region and carbohydrate recognition domain (CRD). Recent studies indicated polymorphisms in neck region of DC-SIGNR were more frequent and associated with susceptibility to HIV-1 infection, whereas no similar trends were observed on DC-SIGN for rare variations of DC-SIGN among the researched subjects in those studies. In our present study, we found 9 genotype variations of DC-SIGN neck region among HIV infected Germans and 3 variations among HIV infected Chinese, respectively. Comparison of those variations between Chinese and Germans reveals strong ethnic discrepancies, and no significant evidence indicates those variations have resistance to susceptibility to HIV-1 infection. PMID- 18424364 TI - In vivo results for interstitial laser application in thyroid gland. AB - OBJECTIVE: Aim of this study was to evaluate the potential of denaturation of hormone active tissue in the thyroid gland by laser induced interstitial thermotherapy (LITT) as a treatment of autonomous hyperthyroidism. MATERIALS AND METHODS: An interstitial thyroid laser application (Nd:YAG 1064 nm, 5W, 2 min) was performed in 5 pigs. During laser application, the laryngeal recurrent nerve was controlled electro-physiologically. Postoperatively, TSH, total T(3) (TT(3)) and free T(4) (FT(4)) were measured regularly. After a follow-up period of up to 6 weeks, pigs were sacrificed and the thyroid glands were evaluated histological. RESULTS: A malfunction of the nerve due to laser treatment was not detected. During the first postoperative week there was a decrease of both FT(4) and TSH whereas TT(3) showed an extreme decline of its plasma levels reaching nearly the detection limit. All values showed a recovery to their initial levels during an interval of 10 days and than increased to levels sometimes higher than baseline. The coagulation zones were demarcated clearly towards normal tissue with increasing fibrosis of the treated areas. CONCLUSION: Interstitial thyroid ablation using a Nd:YAG laser is a minimal invasive, safe and effective procedure. Further evaluation including long term follow-up in humans is needed to confirm these results. PMID- 18424365 TI - Fecal elastase 1 and vitamin D3 in patients with osteoporotic bone fractures. AB - BACKGROUND AND AIMS: The aim of the present study was to clarify if patients with osteoporotic bone fractures have exocrine pancreatic insufficiency, especially reduced fecal elastase 1, connected with lowered serum levels of vitamin D3 that could be relevant for predominant osteoporosis. METHODS: Between October 1999 and September 2001, we investigated on 167 patients with an average age of approx. 69 years suffering from typical osteoporotic bone fractures, as well as 20 healthy controls with an average age of 53 years. A standardized osteodensitometry via dual energy X-ray absorptiometry (DEXA) was performed in all participants. Levels of PTH, 1,25(OH)(2) Vitamin D(3), 25(OH) Vitamin D(3), calcium and phosphate in serum, elastase 1 in feces as well as the body mass index were determined in all patients and controls. RESULTS: In patients 25(OH)D3 was more than 60% and 1,25(OH)(2)D(3) was more than 53% decreased compared to controls. Fecal elastase 1 was lower than the lowest reference of 200 microg/g feces in more than 34% of the patients and it was more than 65% reduced in comparison to healthy controls (fecal elastase 1 patients: 240.7 +/- 96.3 microg/g; controls 694.9 +/- 138.6 microg/g). Separation of the patients in accordance with the elastase 1 contend in feces into four groups (below 100 microg/g, between 100 and 200 microg/g, between 201 and 300 microg/g and above 300 microg/g) resulted in significant variations for 25(OH)D(3), 1,25(OH)(2)D(3), calcium and PTH between these groups (p < 0.01). Furthermore 25(OH)D(3), 1,25(OH)(2)D(3), calcium and PTH correlated significantly with elastase 1 in feces (p < 0.01) the way, that lower fecal elastase 1 was connected with lower levels of the other parameters. BMI shows no relevant differences within the patients or between patients and controls. CONCLUSION: Exocrine pancreatic insufficiency, especially lowered fecal elastase 1, may be much more frequent in patients with osteoporotic bone fractures than suggested so far. Lowered exocrine pancreatic function with lowered fecal elastase 1 seems to be relevant as a reason for reduced levels of circulating vitamin D3 metabolites being an appropriate additional cause for predominant osteoporosis. PMID- 18424366 TI - N-terminal acetylation protects glucagon-like peptide GLP-1-(7-34)-amide from DPP IV-mediated degradation retaining cAMP- and insulin-releasing capacity. AB - Since its discovery glucagon-like peptide-1 (GLP-1) is investigated as a treatment for type II diabetes based on its major function as insulin secretagogue. A therapeutic use is, however, limited by its short biological half life in the range of minutes, predominantly caused via degradation catalyzed by dipeptidyl peptidase IV (DPP-IV). Therefore, we aimed to design a GLP-1 analogue exhibiting resistance against DPP-IV-catalyzed inactivation while retaining its biological activity. By means of matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS) we have studied the stability of the N-terminally blocked new analogue Ac-GLP-1-(7-34)-amide against DPP-IV and compared it with both unblocked GLP-1-(7-34)-amide and the major naturally occurring form GLP-1-(7-36) amide. GLP-1-(7-36)-amide and the C-terminally two amino acid residues shorter GLP-1-(7-34)-amide rapidly generated peptide fragments truncated by the N terminal dipeptide. In contrast, the N-terminal blocked Ac-GLP-1-(7-34)-amide was not degraded in the presence of DPP-IV over a period of at least two hours. Ac GLP-1-(7-34)-amide induced a concentration-dependent increase of intracellular cAMP production and insulin release from rat insulinoma RIN-m5F cells to an extent comparable to that found for the N-terminally unblocked peptides GLP-1-(7 34)-amide and GLP-1-(7-36)-amide. Ac-GLP-1-(7-34)-amide may thus have the potential to act as a new long-acting GLP-1 analogue with significant resistance against DPP-IV and retained biological activity in vitro. Further research is required to investigate whether Ac-GLP-1-(7-34)-amide also exhibits its characteristics in animal models and humans. PMID- 18424367 TI - Ischemic preconditioning improves postoperative outcome after liver resections: a randomized controlled study. AB - BACKGROUND: Clamping of the portal triad (Pringle maneuver) prevents blood loss during liver resection, but leads to liver injury upon reperfusion. Ischemic preconditioning (IP) has been shown to protect the liver against prolonged ischemic injury in animal models. However, the clinical value of this procedure has not yet been established. METHODS: 61 Patients undergoing hepatic resection under inflow occlusion were randomized to either to receive (Group-A n = 30) or not to receive (Group-B n = 31) an IP (10 minutes of ischemia followed 10 minutes of reperfusion). RESULTS: Mean (+/- SD)/ Group-A vs. Group-B. Pringle time of 34 +/- 14 and 33 +/- 12 minutes and the extent of resected liver tissue (2.7 +/- 1.3 vs. 2.7 +/- 1.1 segments) were comparable in both groups. Complications, including death, severe liver dysfunction and biliary leakage occurred in 6 patients of Group-A vs. 14 patients of Group-B (p<0.05). Intraoperative blood loss was significantly lower in Group-A (1.28 +/- 0.91 l vs. 1.94 +/- 0.76 l; p<0.001) with 5 vs. 15 patients requiring transfusions (p<0.01). In a multivariate analysis the duration of the Pringle maneuver (p<0.05) and the absence of preconditioning (p<0.05) were independent predictors for the occurrence of postoperative complications. CONCLUSIONS: IP protects against reperfusion injury, reduces the incidence of complications after hepatic resection under inflow occlusion and is simple to use in clinical practice. PMID- 18424368 TI - Course and therapy of acute liver failure. AB - OBJECTIVES AND METHODS: Despite liver transplantation and advances in intensive care medicine fulminant hepatic failure [FHF] remains a life-threatening condition. Actual observations of the clinical course of these patients are rare. Therefore, we analyzed course of disease and survival in all patients treated for FHF at the University of Bonn between 1998 and 2004 and compared it to the patients treated for FHF during 1992-1997. RESULTS: 35 patients were treated for FHF during this period. FHF was viral induced in 13 patients (HBV n = 11, HAV n = 2), toxic in nine, cryptogenic in eleven and autoimmune and hyperthermia in one patient each. According to London- and/or Clichy criteria 16 patients were transplanted. Four of them died during the first year after transplantation due to infectious and hemorrhagic complications. Three patients died without liver transplantation. All together, 1-year survival was 80%. When compared to patients with FHF analyzed in the period 1992-1997 numbers of patients with FHF in our centre had increased from 16 to 35 patients and 1-year survival improved from 67.5% to 80%. This improved survival was associated with a lower proportion of transplanted patients (45% versus 68%). CONCLUSIONS: These changes reflect advances in therapy of patients with FHF, which enables a greater proportion of patients to survive without the need for transplantation. PMID- 18424369 TI - The epidermis: a sensory tissue. AB - The skin is an efficient barrier which protects our bodies from the external environment but it is also an important site for the perception of various stimuli. Sensory neurones of the peripheral nervous system send many primary afferent fibres to the skin. They pass through the dermis and penetrate the basement membrane to innervate epidermal cells or remain as free endings. Nerve fibres are clearly involved in somatosensation. However, they are not always so numerous, for example in distal parts of the limbs, and some kinds of sensors can be at a distance of hundreds of micrometers from each other. The skin can detect patterns at a very fine and smaller scale, which suggests that nerve terminals are helped by epidermal sensors. All epidermal cells (keratinocytes, melanocytes, Langerhans cells and Merkel cells) express sensor proteins and neuropeptides regulating the neuro-immuno-cutaneous system. Hence, they must play a part in the epidermal sensory system. This review will consider the epidermal components of this forefront sensory system and the stimulations they perceive. The epidermis can be considered a true sensory tissue where sensor proteins and neurone-like properties enable epidermal cells to participate in the skin surface perception through interactions with nerve fibres. PMID- 18424370 TI - Photoprotective effect of a water-soluble extract of Rosmarinus officinalis L. against UV-induced matrix metalloproteinase-1 in human dermal fibroblasts and reconstructed skin. AB - Chronic UV exposure is responsible for long term clinical manifestations such as photoaging and photocancers. One of the major events involved in the development of skin photodamage is up-regulation of matrix metalloproteinase-1 (MMP-1). In this study, the effects of a water-soluble extract of Rosmarinus officinalis (Ro) on the expression of UV-induced MMP-1 were investigated. Using MMP-1 promoter reporter gene constructs, Ro extract was shown to inhibit UV-induced up regulation of MMP-1 gene transcription. The anti-MMP-1 effect was confirmed at the protein level in dermal human fibroblasts exposed either to UVB, UVA or Solar Simulated Radiation. Zymographic analysis on casein and gelatin gels revealed that Ro extract more specifically targeted MMP-1 compared to MMP-2. Using a 3D skin model exposed to Solar Simulated Radiation, anti-MMP-1 activity was confirmed together with a photoprotective effect at the morphological level. Finally the release of cytokines IL1alpha and IL6 which participate in the up regulation of MMP-1 induced by UV exposure could be prevented by the Ro extract. All together, from molecular to tissue level, these results illustrate the ability of the Ro water-soluble extract to inhibit UV-induced MMP-1 and its potential benefits in preventing cutaneous photodamage. PMID- 18424371 TI - Prophylactic effect of oral administration of Lactobacillus johnsonii NCC533 (La1) during the weaning period on atopic dermatitis in NC/NgaTnd mice. AB - Bacterial exposure in infancy may be one of the determinants of atopic dermatitis (AD) morbidity in later life. Some clinical studies have shown that an intake of probiotics reduced the risks of AD in children; however, the timing and duration of administration for the prevention of AD still remain unclear. The aim of this study was to evaluate the effects on AD development of the administration of Lactobacillus johnsonii NC553 (La1) during the weaning period, using an animal model of human AD, NC/NgaTnd mice. La1 suspended in drinking water was administered to 4-week-old NC/NgaTnd mice for 4 weeks. Mice were kept up to 16 weeks of age in an air uncontrolled conventional condition. Clinical skin severity, scratching behaviour, histological features, and production of regulatory or inflammatory cytokines in spleens were analyzed. The results indicated that oral administration of La1 suppressed exacerbation of the clinical severity of dermatitis when compared to the controls. Scratching duration, which is the most important cause of skin damage, was also suppressed in mice fed with La1. La1 supplementation also suppressed epidermal hyperplasia and infiltration of inflammatory cells in skin. This study showed that exposure to La1 from the early stages might be beneficial to reduce the exacerbation of AD in children at high-risk of allergy. PMID- 18424372 TI - Differential effects of allergens and irritants on early differentiating monocyte derived dendritic cells. AB - Phenotypic modifications induced by contact allergens on monocyte-derived dendritic cells (MoDC) have been proposed as an in vitro alternative method to discriminate potential sensitizers from irritants. However, the sensitivity of the assay remains controversial. In all the studies reported so far, DC treatment with chemicals was carried out after 5 to 6 days of monocyte culture. Here, we first determined the dynamic range of expression of differentiation and activation markers on human MoDC cultured in the presence, or absence, of TGFbeta. At day three of culture, most monocytes have already differentiated into CD1a+/CD14- DC and, in the presence of TGFbeta, they expressed CD40, CD54 or CD86 antigens with lower fluorescence intensity than 5 day-cultured MoDC. Treatment of 3-day cultured TGFbeta-MoDC with all the tested strong and moderate sensitizers, i.e. NiSO(4), DNCB, balm of Peru, isothiazolinone and cinnamic aldehyde, at non toxic concentrations, induced significant phenotypic changes, whereas the irritant SLS had no effect. However, a large variability was observed in the number and nature of the modified antigens, according to the chemical and the experiments. This implies that many surface antigens must be analyzed and many experiments carried out to use this assay as an alternative screening method for contact sensitizers. PMID- 18424373 TI - Imiquimod in mycosis fungoides. AB - Imiquimod is a topically active imidazoquinoline immunomodulator agent. It works as an indirect antiviral and antitumoral and stimulates the production of INF alpha and various other cytokines. We assayed topical imiquimod in treating early stages of mycosis fungoides. We applied imiquimod 5% cream in four patients with multi-treatment resistant plaques of MF (stages IA and IIB). We applied it on one patient in association with systemic INFalpha-2a. We observed a complete clinical clearance of the lesions in all four patients. In three cases we achieved a complete histopathological clearance and in one case a partial histopathological clearance. The patient treated with imiquimod and systemic INFalpha-2a showed the most spectacular improvement with a rapid total response. We ascribe this improvement to a synergic effect of imiquimod and systemic INFalpha-2a treatment. Before the introduction of imiquimod, this patient had been treated for 2 years with systemic INFalpha-2a alone, without any evidence of clinical response. Imiquimod could be an effective therapy for early-stage disease of CTCL, used alone or in combination with systemic immunomodulatory therapy. PMID- 18424374 TI - Study of persistence and recurrence rates in 106 patients with condyloma and intraepithelial neoplasia after CO2 laser treatment. AB - Our aim was to evaluate remission and relapse rates and the number of laser sessions necessary for treatment. Among the relapses observed, we sought to differentiate between the persistence and recurrence of an HPV-induced lesion. This retrospective study was performed in patients, immunocompetent or not, treated with CO2 laser for condylomatous or neoplastic anogenital lesions by the same operator over a period of 12 months. 106 treated patients were followed for 6 months. Three groups of patients were analysed: HIV(+) patients, patients with therapeutic immunosuppression (ImST) and immunocompetent patients (ImC). Twenty seven (25.5%) patients presented with high-grade intraepithelial neoplasms (IEN III). IEN III lesions were more common in the HIV(+) group than in immunocompetent patients (47.4% versus 20.2%, p = 0.015). The development of HPV induced lesions at several sites on the body was also more common in HIV(+) patients. Post-laser controls at one month demonstrated a clinical absence of HPV induced lesions in 81.2% of cases, recurrence in 12.6% of cases and persistence in 6.6% of cases. Remission rates at one month did not differ significantly between the three groups. 93% of patients in remission at one month were still in remission at three months. IEN III neoplasms in remission at one month remained so at six months. ImC and ImST patients presented more frequently with recurrence than persistence, when compared with HIV(+) patients. At six months, 83% of patients were in remission after 1.4 laser treatments. The excision of HPV induced anogenital lesions using CO2 laser remains an efficient treatment, even if it needs to be repeated if lesions recur or persist. CO2 laser treatment under colposcopic guidance can achieve remission in both immunocompromised and non compromised patients with longstanding lesions. PMID- 18424375 TI - Treatment of pemphigus vulgaris with mycophenolate mofetil as a steroid-sparing agent. AB - Pemphigus vulgaris is a rare autoimmune blistering disease. Estimation of the incidence in Iran is one patient per 100,000 of the population per year. Mycophenolate mofetil is an immunosuppressive drug and successful treatment of pemphigus vulgaris and bullous pemphigoid has been reported with it, in combination with high dose prednisone, or as monotherapy. The present study describes our experience of the adjuvant use of mycophenolate mofetil in the management of 31 patients with pemphigus vulgaris as an initial treatment. We evaluated the efficacy and safety of mycophenolate mofetil combined with prednisolone in this cohort. We also assessed the relationship between the demographic indices/disease severity factors, and the failure of this treatment. In this study, mycophenolate mofetil was of definite benefit in 21 cases (67.7%). Generalized forms; patients with higher sum of the clinical scores at presentation; severe involvement of the groin; chest; face and limbs and those who had nail dystrophy also appeared to have poorer responses. When we excluded patients with generalized forms, only four patients were included in the failure group and the response rate reached 83.3%. It can be concluded that, except for generalized diseases, mycophenolate mofetil can be used safely and effectively in patients with pemphigus vulgaris as a first line, steroid sparing agent. PMID- 18424376 TI - Increased frequency of self-reported parasomnias in patients suffering from vitiligo. AB - Vitiligo is a skin disorder of unknown aetiology, affecting 0.1-2% of the general population. The aim of the present study was to investigate its relationship with sleep disorders, especially parasomnias. Two hundred and sixteen individuals were examined. Among them, 116 were suffering from vitiligo, 52 from other dermatological diseases and 48 were healthy subjects, serving as a control group. An inventory including items related to sleep disorders from childhood and adolescence was used. The study was focused specifically on parasomnias. Patients suffering from vitiligo reported a significantly higher occurrence of sleepwalking, nocturnal enuresis, night illusions, sleep terrors and nightmares than that of the control group, prior to the manifestation of the disease. Patients suffering from other dermatological diseases only reported significantly more often nightmares and nocturnal enuresis compared to the control group. A relationship between parasomnias during early life and later development of vitiligo was detected. This finding supports the hypothesis that neural mechanisms involving monoaminergic systems (especially the serotoninergic one) may potentially be involved in the aetiopathology of vitiligo. PMID- 18424377 TI - Is facial involvement a sign of severe psoriasis? AB - Facial involvement in psoriasis receives little attention in standard descriptions of the disease. To our knowledge there has been only one comparison study on the severe of psoriasis between patients with and without facial involvement. The purpose of this study was to define the prevalence and characteristics of facial involvement, and to compare the severity of psoriasis between patients with and without facial involvement. A total of 120 consecutive patients with psoriasis seen in our polyclinic between January 2005 and May 2005 were enrolled in this study. They were categorized into patients with and without facial psoriasis. The age of onset, family history, nail and joint involvement, associated skin or systemic disease, history of phototherapy or systemic therapy, extent of involvement, the effect of external factors and Psoriasis Area and Severity Index (PASI) scores were recorded. Patients who had PASI score lower than 5 were excluded from the study. Patients with facial psoriasis frequently report early onset, or long duration, and tend to have more extensive disease and more intensive treatment. The patients with facial psoriasis have more positive family histories and Koebner responses. The nail, joint involvement and pruritus did not appear to influence the presence of facial psoriasis. Facial involvement in psoriasis is much more common than generally appreciated. This study demonstrates the significance of facial involvement in psoriasis. In view of its significance as a marker of severe disease, extra care must be taken during treatment to ensure clearance from this site. PMID- 18424378 TI - Facing psoriasis and atopic dermatitis: are there more similarities or more differences? AB - Atopic dermatitis (AD) and psoriasis vulgaris (Pso) represent the most frequent chronic inflammatory skin diseases. It has been assumed for a long time that these diseases have a completely different background. Recent findings about the genetic, epidemiologic and pathophysiologic factors of both diseases have remarkably improved our knowledge about the complex mechanisms underlying AD and Pso. Beyond that, in view of these findings, the question arises, which similarities and differences between AD and Pso exist. In order to address this point, we provide an overview about the current knowledge in the field of AD and Pso. PMID- 18424379 TI - Patient's perspective: an important issue not to be overlooked in assessing acne severity. AB - Acne has significant negative effects on an individual's psychosocial functions. There is not always a correlation between the severity of acne and its impact on quality of life. Our objective was to evaluate the correlation between quality of life scales and both the physician's and patient's assessments of acne severity and to find out which quality of life scale is more sensitive to changes in acne severity by using generic and acne specific scales. One hundred and twenty acne patients were enrolled. The physician's assessment of acne severity was made by means of Global Acne Grading System. Patients evaluated their acne severity on a 10-point Likert-type scale. Quality of life was measured by the Turkish version of Acne Quality of Life (AQOL) scale and Short Form-36 (SF-36). One hundred and seven patients answered the questions of the AQOL scale completely and were included in the statistical analysis. The AQOL scale did not correlate with the physician's assessment of acne severity whereas there was a correlation between patients' self assessments and AQOL scale. No correlation was found between SF-36 and either the physician's or patients' own assessments. Assessment of acne should not be limited to objective acne severity measures but also include patients' self assessments and acne specific quality of life scales. These measures, which provide a better understanding of patients' perception of severity, are important tools when taking treatment decisions. PMID- 18424380 TI - Hair follicle nevus - A dermoscopic approach. AB - We report the case of a 26-year-old man who presented with small soft nodules with tiny hairs that had been present on his nose since childhood. The nodules were initially diagnosed as melanocytic nevi. However, dermoscopy showed many uniform hair follicles and an interfollicular 'pseudo-pigment network' in the nodules. Histologically, many well-differentiated hair follicles and sebaceous glands were seen in the dermis. Serial sectioning revealed neither central cysts nor a central canal. We therefore diagnosed this case as hair follicle nevus. Dermoscopy is now widely used as a non-invasive, in vivo technique for the diagnosis of pigmented skin lesions. Hair follicle nevus is a very rare disease and this is the first report to demonstrate the manifestation of this clinical entity by dermoscopy. PMID- 18424381 TI - Cutaneous lymphoid hyperplasia and reactive lymphadenopathy induced by hepatitis B vaccination. PMID- 18424382 TI - Subcutaneous juvenile xanthogranuloma. PMID- 18424384 TI - Cutaneous sarcoidosis presenting with pinhead-sized papules. PMID- 18424383 TI - Severe liver dysfunction due to bosentan in a patient with mixed connective tissue disease. PMID- 18424385 TI - Acrodermatitis enteropathica in a full-term exclusively breast-fed infant. PMID- 18424386 TI - Simultaneous occurrence of reactive angioendotheliomatosis and leukocytoclastic vasculitis in a patient with periodontitis. PMID- 18424387 TI - Exudative erythema multiforme induced by hydroxyzine. PMID- 18424388 TI - Paraneoplastic dermatomyositis triggered and exacerbated by oral 5-fluorouracil administration. PMID- 18424389 TI - Accentuated Blaschko-linear manifestation of psoriasis. PMID- 18424390 TI - Erythema gyratum repens as the initial manifestation of lung cancer. PMID- 18424391 TI - Minocycline in combination with mycophenolate mofetil in oral mucous membrane pemphigoid. PMID- 18424392 TI - Progressive nodular histiocytosis - A five-year follow up. PMID- 18424393 TI - Inflammation of actinic keratoses with capecitabine therapy for colon cancer. PMID- 18424394 TI - What could be expected after fifteen years of amiodarone therapy? PMID- 18424395 TI - Angiomatoid lesions revealing an atrial myxoma. PMID- 18424396 TI - Leg ulcer and bacteremia due to Cedecea davisae. PMID- 18424397 TI - Erythema annulare centrifugum associated with herpes zoster. PMID- 18424398 TI - Hand warts successfully treated with topical 5-aminolevulinic acid and intense pulsed light. PMID- 18424399 TI - Coexistence of congenital malalignment of the great toenails with ocular melanocytosis. PMID- 18424400 TI - DRESS in a patient taking a dietary supplement containing phenobarbital purchased over the internet. PMID- 18424401 TI - European Dermatology Forum: skin diseases in Europe. Skin diseases with a high public health impact: chronic venous insufficiency. PMID- 18424402 TI - European Dermatology Forum: skin diseases in Europe. Skin diseases with a high public health impact: scleroderma. PMID- 18424403 TI - European Dermatology Forum: skin diseases in Europe. Skin diseases with a high public health impact: Epidermolysis bullosa. PMID- 18424404 TI - European Dermatology Forum: skin diseases in Europe. Skin diseases with a high public health impact: toxic epidermal necrolysis and Stevens-Johnson syndrome. PMID- 18424405 TI - European Dermatology Forum: skin diseases in Europe. Skin diseases with a high public health impact: HIV infection. PMID- 18424407 TI - [Recent development of metabonomics and its applications in clinical research]. AB - In the post-genomic era, systems biology is central to the biological sciences. Functional genomics such as transcriptomics and proteomics can simultaneous determine massive gene or protein expression changes following drug treatment or other intervention. However, these changes can't be coupled directly to changes in biological function. As a result, metabonomics and its many pseudonyms (metabolomics, metabolic profiling, etc.) have exploded onto the scientific scene in the past several years. Metabonomics is a rapidly growing research area and a system approach for comprehensive and quantitative analysis of the global metabolites in a biological matrix. Analytical chemistry approach is necessary for the development of comprehensive metabonomics investigations. Fundamentally, there are two types of metabonomics approaches: mass-spectrometry (MS) based and nuclear magnetic resonance (NMR) methodologies. Metabonomics measurements provide a wealth of data information and interpretation of these data relies mainly on chemometrics approaches to perform large-scale data analysis and data visualization, such as principal and independent component analysis, multidimensional scaling, a variety of clustering techniques, and discriminant function analysis, among many others. In this review, the recent development of analytical and statistical techniques used in metabonomics is summarized. Major applications of metabonomics relevant to clinical and preclinical study are then reviewed. The applications of metabonomics in study of liver diseases, cancers and other diseases have proved useful both as an experimental tool for pathogenesis mechanism re-search and ultimately a tool for diagnosis and monitoring treatment response of these diseases. Next, the applications of metabonomics in preclinical toxicology are discussed and the role that metabonomics might do in pharmaceutical research and development is explained with special reference to the aims and achievements of the Consortium for Metabonomic Toxicology (COMET), and the concept of pharmacometabonomics as a way of predicting an individual's response to treatment is highlighted. Finally, the role of metabonomics in elucidating the function of the unknown or novel enzyme is mentioned. PMID- 18424408 TI - [Genome-wide association study on complex diseases: study design and genetic markers]. AB - Genome-wide association study used to be a dream of geneticists years ago, but now it came true. Since the first paper reported the finding of genetic variation contributing to human age-related macular degeneration by genome-wide association study in 2005, a numbers of whole genome studies have been published. The present paper reviewed some common comments in whole genome association study on complex diseases, including achievements of genome-wide association studies on complex traits or diseases, principles of study design, selection of genetic marker in genome, and comparisons of different commercial products for whole genome association study. Finally a newly defined genetic variation, copy number variation, was briefly introduced. This paper also summarized the shortcomings of current genome-wide association studies and perspectives of its future. PMID- 18424409 TI - [Exploration of carcinogenesis based on tree models using CGH data]. AB - Comparative genomic hybridization (CGH) can detect chromosomal deletions and amplifications of tumors, and various laboratories and public databases have accumulated a large number of CGH data, providing the opportunity to analyze the molecular mechanism of tumorigenesis in the whole genome. Tree models are generally used to study the history of biological formation and evolution in the field of bioinformatics, and evolutionary relationships between species are usually represented using phylogenetic tree. Tree models are also powerful bioinformatics tools to analyze CGH data and explore carcinogenesis. Two common tree models, the branching tree and the distanced-based tree, as well as their basic principles, methods are introduced detailedly, several technical problems in construction of tree models are discussed, and their applications in cancer research are reviewed systematically in this paper. As a generalization of single path linear model, tree models can more accurately conclude multigene, multistep, multipathway process of tumorigenesis, exploring the molecular mechanism of tumorigenesis from different angels. Apart from CGH data, tree models can be used to analyze various types of data, including high-resolution data (e.g., array-CGH data). PMID- 18424410 TI - [Genetic polymorphism for genes of alcohol dependence]. AB - Alcohol dependence is a complex disorder which is influenced by physiological, psychological, environmental factors, individual inheritance and so on. Several candidate genes associated with alcohol dependence risk have been identified. The review focuses on several related genes that control alcohol metabolism such as alcohol dehydrogenase, aldehyde dehydrogenase, cytochrome P450 2E1 and regulate neurotransmission such as catechol-O-methyltransferase, dopamine receptors D2 and D4, and mu opioid receptor. PMID- 18424411 TI - [KISS-1/GPR54 genes and their role in reproduction]. AB - KISS-1 gene and its receptor gene GPR54 play key roles in the initiation of puberty onset. The peptide product of the KiSS-1 gene, Kisspeptins stimulate gonadotrophins release to initiate puberty through the expression of GPR54 gene in the brain. So the level of KISS-1 and GPR54 mRNA in hypothalamus was very high on the onset of puberty. The expression of KISS-1gene was regulated by steroid hormone in different nuclei within the forebrain to control the reproduction in puberty. Loss of function mutations of GPR54 gene could cause idiopathic hypogonadotropic hypogonadism (IHH) and gonadotrophin-dependant premature puberty. This review also introduced the structure, expression, homology comparison, polymorphism of KISS-1 and GPR54 genes and their interrelation with other regulators of reproduction. PMID- 18424412 TI - [DNA methylation regulating factors in plants]. AB - DNA methylation is an important epigenetic modification and multiple factors interact to regulate the establishment and maintenance of DNA methylation in plant genome. Different methylation sites require different cytosine methyltransferases, which contribute to the modification of chromatin structure and mediate epigenetics with chromatin remodeling enzymes and histone modifying factors. DNA glycosylases can remove DNA methylation and alleviate silencing. The functions and interactions of DNA methylation regulating factors, the establishment, maintenance and removement mechanisms of DNA methylations are reviewed in this paper. PMID- 18424413 TI - [Mutation screening of MITF gene in patients with Waardenburg syndrome type 2]. AB - Warrgenburg syndrome type 2 (WS2) is the most common autosomal dominantly inherited syndrome with hearing loss. MITF (microphthalmia associated transcription factor)is a basic-helix-loop-helix-luecine zipper (bHLHZip) factor which regulates expression of tyrosinase, and is involved in melanocyte differentiation. Mutations in MITF associated with WS2 have been identified in some but not all affected families. Here, we report a three-generation Chinese family with a point mutation in the MITF gene causing WS2. The proband exhibits congenital severe sensorineural hearing loss, heterochromia iridis and facial freckles. One of family members manifests sensorineural deafness, and the other patients show premature greying or/and freckles. This mutation, heterozygous deletion c.639delA, creates a stop codon in exon 7 and is predicted to result in a truncated protein lacking normal interaction with its target DNA motif. This mutation is a novel mutation and the third case identified in exon 7 of MITF in WS2. Though there is only one base pair distance between this novel mutation and the other two documented cases and similar amino acids change, significant difference is seen in clinical phenotype, which suggests genetic background may play an important role. PMID- 18424414 TI - [Sequence polymorphism of mtDNA HVR Iand HVR II of Oroqen ethnic group in Inner Mongolia]. AB - Venous blood samples from 50 unrelated Oroqen individuals living in Inner Mongolia were collected and their mtDNA HVR I and HVR II sequences were detected by using ABI PRISM377 sequencers. The number of polymorphic loci, haplotype, haplotype frequence, average nucleotide variability and other polymorphic parameters were calculated. Based on Oroqen mtDNA sequence data obtained in our experiments and published data, genetic distance between Oroqen ethnic group and other populations were computered by Nei's measure. Phylogenetic tree was constructed by Neighbor Joining method. Comparing with Anderson sequence, 52 polymorphic loci in HVR I and 24 loci in HVR II were found in Oroqen mtDNA sequence, 38 and 27 haplotypes were defined herewith. Haplotype diversity and average nucleotide variability were 0.964+/-0.018 and 7.379 in HVR I, 0.929+/ 0.019 and 2.408 in HVR II respectively. Fst and dA genetic distance between 12 populations were calculated based on HVR I sequence, and their relative coefficients were 0.993(P < 0.01). A phylogenetic tree was constructed based on genetic distances and included Oroqen, Taiwan and South Han population in a clade, which indicated near genetic relation between them, and far relation with northern Han, Mongolian and other foreign populations. The genetic polymorphism of mtDNA HVR I and HVR II in Oroqen ethnic group has some specificities compared with that of other populations. These data provide a useful tool in forensic identification, population genetic study and other research fields. PMID- 18424415 TI - [Amplification and expression of human Artemin cDNA]. AB - The cDNA fragment encoding human Artemin was amplified by RT-PCR with human fetal brain RNA as template. Result of sequencing showed that the homology of nucleotides is 99.7% between the amplified human Artemin cDNA and the reported one (GenBank accession No. AF115765) and the homology of amino acids is 100%. The prokaryotic expressing vector pGEX-6p-1-hART carrying the amplified DNA fragment was obtained by ligation with the plasmid pGEX-6p-1. The expression of recombinant human Artemin fusion protein in E.coli was analyzed by SDS polyacarylamide gel electrophoresis. Result demonstrated that the recombinant protein, mostly found in inclusion bodies, accounted for 18.32% of the total bacterial lysate. The inclusion body was dissolved and renatured by the oxidoreduction system, and the recombinant protein is analyzed by Western blotting. Human Artemin cDNA was amplified and the recombinant protein was expressed in vitro successfully. PMID- 18424416 TI - [Association of SNPs in the promoter of MMP-2 and TIMP-2 genes with epithelial ovarian cancer]. AB - The association between single nucleotide polymorphisms (SNPs) in the promoter region of MMP-2 and TIMP-2 genes and the risk of epithelial ovarian cancer was investigated. MMP-2 C-1306T, C-735T and TIMP-2 G-418C SNPs were genotyped by polymerase chain reaction-restrictive fragment length polymorphism (PCR-RFLP) analysis in 246 patients with epithelial ovarian cancer and 324 healthy women as control. Results showed no significant difference between the patient and control groups in allele or genotype distributions of MMP-2 C-1306T (P=0.55 and P=0.42). However, the frequencies of the C allele and the C/C genotype of the MMP-2 C-735T were significantly higher in ovarian cancer patients (80.7% and 66.7%) than those in healthy controls (75.5% and 55.9%). Compared with the T/T+C/T genotypes, the C/C genotype significantly increased the risk of ovarian cancer (OR=1.58, 95%CI=1.12-2.23). Stratification analysis showed that subjects carrying C/C genotype were significantly associated with the risk of endometrioid ovarian cancer and with ovarian cancer in subjects that were 50 or older, with odds ratio at 1.69 (95%CI=1.03-2.79) and 1.71 (95%CI=1.14-2.57), respectively. Haplotype analysis showed that the frequencies of four haplotypes (T(-1306)-T(-735), T( 1306)-C(-735), C(-1306)-T(-735) and C(-1306)-C(-735)) of MMP-2 C-1306T and C-735T were not significantly different between the patient and control groups (P=0.24). The allele and genotype frequencies of TIMP-2 G-418C were not significantly different between the patient and control groups (P=0.33 and P=0.47). But TIMP-2 418G/G genotype was associated with a trend for endometrioid ovarian cancer by stratification analysis according to histological subtypes (OR=1.62, 95%CI=0.94 2.78). Thus, the study suggested that the C/C genotype of the C-735T SNP in the promoter region of MMP-2 gene may be a potential risk factor for epithelial ovarian cancer, but the C-1306T SNP may have no association with the risk of epithelial ovarian cancer. The TIMP-2 G-418C SNP may be associated with the risk of different histological subtypes of epithelial ovarian cancer. PMID- 18424417 TI - [Study on the relationships between polymorphisms of CXCR2 gene and milk quality and mastitis of dairy cow]. AB - PCR-SSCP was applied to analyze the polymorphisms of CXCR2 gene and its relationships with milk quality and mastitis in 160 cattle samples including Holstein dairy cow, Simmental dairy cow and Tongjiang cattle. The results showed that there were 3 SNP loci in CXCR2 gene, which located at 684 bp, 777 bp, 861 bp, respectively. Five alleles (A, B, C, E and F) were determined based on the 3 SNP loci. The genotypes for each locus were BC and CC, AA and AB, CC and BC at 685 bp, 777 bp and 861 bp, respectively. Results also demonstrated that genotypes BC, CC and FF were probably relevant with mastitis, but AA, BB and EE may resist the mastitis of cattle. The genotypes AA, AB and EE may have better milk quality (extremely significant difference or significant difference) than other genotypes. PMID- 18424418 TI - [Analysis on molecular evolution of MC1R gene in dog]. AB - Melanocortin 1 receptor (MC1R) gene regulates pigment synthesis in mammals, and therefore is regarded as an important candidate gene for dog coat color. Based on MC1R amino acids and cDNA sequences of 10 vertebrate animals released by NCBI, molecular evolution of dog MC1R gene was analyzed with bioinformatic software and internet resource. Results showed that 10 vertebrate animals were divided into two major groups, a compact group A (7 mammals) and an incompact group B (chicken, zebrafish and fugu). This phylogenetic tree was consistent with putative evolutionary relationship within these 10 species. Positive selection was detected during the evolutionary process of dog (also cat and pig) from cattle by PAML branch model (omega = 90.8177), and five amino acids of 2V, 25E, 184N, 197V and 314L of dog MC1R were predicted under positive selection by site model. Comparative linkage analysis of chromosome showed that "ZFP276-MC1R-GAS8" linkage group was conservative in human, chimpanzee, chicken and dog. PMID- 18424419 TI - [SNP discovery by F-CSGE in the coding region of mitochondrial DNA in wild house mice from Shanghai suburb]. AB - This study was performed to discover SNPs for genetic polymorphism analysis of mitochondrial DNA from wild house mice. Universal primer florescent PCR, fluorescence-based conformation sensitive gel electrophoresis (F-CSGE) and DNA sequencing were conducted to analyze the coding region of mitochondrial DNA. Different types of unknown mutations were recorded by variable F-CSGE patterns without false positive. Twenty-four SNPs, sixteen of which were first discovered in the coding region of mitochondrial DNA, were found in 64 wild house mice from 4 districts in Shanghai. Therefore, F-CSGE was proved to be powerful technique for SNP discovery in the coding region mitochondrial DNA. The novel SNPs can be used as molecular markers to analyze population structure and genetic polymorphisms of the wild house mice in Shanghai. PMID- 18424420 TI - [Mutation rate and pattern of microsatellites in gynogenetic silver crucian carp (Carassius autatus gibeblio)]. AB - The natural gynogenetic triploid silver crucian carp (Carassius autatus gibeblio Bloch) provides a good system for studying evolutional genetics of the unisexual and polyploidy vertebrate. Microsatellites are abundant across genomes and show high levels of polymorphism and mutational rate, so they have been widely used for studying evolutional biology. In this study, the mutation rate and pattern at 33 microsatellite loci of silver crucian carp were investigated. As a result, it was found that the only one of 22 offspring had 18 mutant alleles at 15 microsatellite loci. The overall mutation rate of the 33 loci was 1.16x10( 2)/locus/generation (95% confidence interval 6.87x10(-3) and 1.83x10(-2)). The mutation rate in the gynogenetic triploid silver crucian carp was obviously higher than other fish, which was closely related to the transitional phase of parthenogenesis and gamogenesis in the natural gynogenetic fish. The repeat numbers had more than 10 times at 13 loci of the mutant alleles, and there was no obviously different in the mutant rate between the 11 compound microsatellite loci (1.31x10(-2) )and the 21 perfect microsatellite loci(1.00x10(-2)) (P = 0.67). The mutant rate had affinity with repeat numbers instead of repeat types and GC content in flanking sequences of microsatellite. The mutation pattern of silver crucian carp was very complexional, as well as some loci did not follow the Stepwise Mutation Model. PMID- 18424421 TI - [SSR mapping of stripe rust resistance gene from Ae. tauschii]. AB - A dominant wheat stripe rust resistance gene, temporarily designated as YrY201, was identified in an accession Y201 of Aegilops tauschii. By bulk segregation analysis, three microsatellite markers Xgwm273b, Xgwm37 and Wmc14 were found to be linked to YrY201 with genetic distance of 11.5, 5.8 and 10.9 cM , respectively. According to the locations of the linked markers, the resistance gene was located on chromosome 7DL. Based on the chromosomal location and the resistance pattern of the gene, we proposed that YrY201 was a novel stripe rust resistance gene, and could be selected by marker-assisted selection. PMID- 18424422 TI - [Overexpression of cotton, vacuolar H+-ATPase subunit C promotes cell elongation and regulates stress tolerance in fission yeast]. AB - Cotton (Gossypium hirsuturm L.) vacuolar H+-ATPase plays an important role in fiber cell elongation through regulating the turgor pressure, which is involved in polarity enlargement of fiber cell. Arabidopsis vacuolar H+-ATPase subunit C (DET3) regulates the activity of vacuolar H+-ATPase and subsequently promotes cell elongation. To rapidly characterize the function of cotton vacuolar H+ ATPase subunit C (GhDET3) and speculate the role of GhDET3 in cotton fiber growth and development, we constructed an expressing vector for fission yeast and introduced it into fission yeast. The results revealed that overexpression of GhDET3 was capable of promoting fission yeast cell elongation and enhancing the tolerance to high NaCl and high pH stresses. These results illuminated that the effects of GhDET3 were important for the activity of vacuolar H+-ATPase and suggested that GhDET3 plays a crucial role in fiber cell elongation. PMID- 18424423 TI - [Application of kanamycin in transgenic mustard(Brassica juncea Coss.)]. AB - In order to find the best screening kanamycin concentration in the genetic transformation of mustard (Brassica juncea Coss.), the seedling cotyledons of mustard were placed on bud-induced media supplemented with different kanamycin concentrations. The bud differentiation of explants was totally inhibited when the kanamycin concentration was greater than 30 mg/L. The seeds of mustard were placed on germination media supplemented with different concentrations of kanamycin. All the seedlings were white when kanamycin concentration was higher than 200 mg/L. The leaves of mustard in field were smeared with the solutions including different concentrations of kanamycin. The treated leaves showed white when kanamycin concentration was over 200 mg/L. To study the segregation of the alien gene in transgenic mustard offspring, the transgenic mustard seeds and the leaves of the transgenic mustard offspring using npt-gene as assistant selection marker were treated with 200 mg/L kanamycin, and the results were analyzed by chi square test. The segregation ratio in the offspring of 4 transgenic lines with single copy of transgene agreed with a ratio of 3:1. The segregation ratio in offspring of the one transgenic line with double copies was agreed with a ratio of 3:1, and the segregation ratio in offspring of the other transgenic line with double copies was agreed with ratios of 3:1 and 15:1 simultaneously. It is indispensable to thoroughly study the insert of the double copies of transgenes in transgenic mustard. PCR technology was used to confirm the above detection methods. It is concluded that applying kanamycin to screen transgenic mustard offspring is feasible. PMID- 18424424 TI - [Cloning of the gene encoding a key enzyme involved in production of glycerol in Candida glycerinogenes]. AB - Candida glycerinogenes WL2002-5, an excellent glycerol producer, has been used for industrial scale fermentation of glycerol by an aerobic process. However, our knowledge about glycerol biosynthesis at the molecular level and genetic background of this yeast species lags far behind those of model yeasts such as Saccharomyces cerevisiae et al. In this report, inverse primers, in conjunction with degenerated primers, were used to amplify the NAD+-dependent glycerol 3 phosphate dehydrogenase (GPD) encoding gene from C. glycerinogenes. The completed nucleotide sequence of the coding, as well as flanking genomic regions was determined (GenBank accession No. EU186536). DNA sequence analysis revealed the present of the open reading frame (ORF) of 1,167 bp, encoding a polypeptide with 388-amino-acid with a molecular mass of 42,695 Da. The CgGPD did not exhibit significant sequence similarity with others described in other eukaryotic systems by comparative analysis. However, it consisted of two typical functional domains which belong to almost all eukaryotic GPDs: a co-enzyme binding domain in the N terminal, and a catalytic domain. Moreover, some relevant features involved in initiation, regulation and stress response element of gene transcription were observed in the nucleotide sequence of the 5'-non-coding regions. Heterologous expression of CgGPD gene in S. cerevisiae improved its glycerol production significantly. In conclusion, the functional CgGPD has been cloned and identified successfully from C. glycerinogenes genome. PMID- 18424425 TI - [Agrobacterium tumefaciens-mediated transformation of the pathogen of Botrytis cinerea]. AB - Employing gfp as a reporter gene and hygromycin gene (hph) as a selection marker, the recombinant vector pKPG was constructed and transformed into fresh conidia of Botrytis cinerea via Agrobacterium tumefaciens. Transformants were identified by PCR analysis of gfp and hph cassette, green fluorescence observation with microscope and Southern hybridization. Results confirmed that target genes were successfully integrated into the genome of Botrytis cinerea. PMID- 18424426 TI - [Establishment and preliminary application of multiplex PCR for detecting the microsatellite markers in silkies]. AB - In order to obtain the stable PCR combinations for silkies genetic analysis, multiplex PCR was used and its reaction condition was optimized. Five combinations of multiplex PCR with good effects were obtained from eighteen microsatellite markers. Three of them were quadruplex-PCRs, and two were triplex PCRs. The products of multiplex PCR were further combined into three sets for the electrophoresis on ABI 3100-Avant Genetic Analyzer. The results suggested that the eighteen microsatellite markers could be successfully applied to the silkies genetic analysis. PMID- 18424427 TI - Trophoblast invasion: the role of intracellular cytokine signalling via signal transducer and activator of transcription 3 (STAT3). AB - Trophoblast cells display a very unique capability: they physiologically invade into the surrounding tissue. This capability is widely associated with tumours, and, indeed, the invasive behaviour of both is rather similar. The imposing difference is that trophoblast cell invasion is temporally and locally controlled in contrast to unlimited tumour invasion. It initiates immediately after embryo implantation into the endometrium. Parallel to tumours, trophoblasts secrete proteases, such as matrix metalloproteinases, which dissolve the extracellular matrix and the surrounding tissue. Thereby, these proteases prepare and allow true invasion of trophoblasts. The invasive capacities of trophoblasts are positively and negatively regulated by numerous cytokines including leukaemia inhibitory factor (LIF), interleukin-6, hepatocyte growth factor, granulocyte macrophage-colony stimulating factor and others. They interact via specific receptors with the trophoblast cells, in which they activate intracellular signalling cascades. These will then induce expression of invasion relevant genes. One of these signalling pathways is the Janus kinase/signal transducers and activators of transcription (STAT) pathway. Especially phosphorylated STAT3 enhances invasiveness of tumours and trophoblast cells, where it is mainly activated by LIF. One of its most efficient physiological antagonists is suppressor of cytokine signalling 3. The balance of these two intracellular molecules seems to be a key regulator of tumour and trophoblast invasion. PMID- 18424428 TI - Are patients with skin cancer at lower risk of developing colorectal or breast cancer? AB - Ultraviolet exposure may reduce the risk of colorectal and breast cancer as the result of rising vitamin D levels. Because skin cancer is positively related to sun exposure, the authors hypothesized a lower incidence of breast and colorectal cancer after skin cancer diagnosis. They analyzed the incidence of colorectal and breast cancer diagnosed from 1972 to 2002 among 26,916 Netherlands skin cancer patients (4,089 squamous cell carcinoma (SCC), 19,319 basal cell carcinoma (BCC), and 3,508 cutaneous malignant melanoma (CMM)). Standardized incidence ratios were calculated. A markedly decreased risk of colorectal cancer was found for subgroups supposedly associated with the highest accumulated sun exposure: men (standardized incidence ratio (SIR) = 0.83, 95% confidence interval (CI): 0.71, 0.97); patients with SCC (SIR = 0.64, 95% CI: 0.43, 0.93); older patients at SCC diagnosis (SIR = 0.59, 95% CI: 0.37, 0.88); and patients with a SCC or BCC lesion on the head and neck area (SIR = 0.59, 95% CI: 0.36, 0.92 for SCC and SIR = 0.78, 95% CI: 0.63, 0.97 for BCC). Patients with CMM exhibited an increased risk of breast cancer, especially advanced breast cancer (SIR = 2.20, 95% CI: 1.10, 3.94) and older patients at CMM diagnosis (SIR = 1.87, 95% CI: 1.14, 2.89). Study results suggest a beneficial effect of continuous sun exposure against colorectal cancer. The higher risk of breast cancer among CMM patients may be related to socioeconomic class, both being more common in the affluent group. PMID- 18424429 TI - Understanding sequelae of injury mechanisms and mild traumatic brain injury incurred during the conflicts in Iraq and Afghanistan: persistent postconcussive symptoms and posttraumatic stress disorder. AB - A cross-sectional study of military personnel following deployment to conflicts in Iraq or Afghanistan ascertained histories of combat theater injury mechanisms and mild traumatic brain injury (TBI) and current prevalence of posttraumatic stress disorder (PTSD) and postconcussive symptoms. Associations among injuries, PTSD, and postconcussive symptoms were explored. In February 2005, a postal survey was sent to Iraq/Afghanistan veterans who had left combat theaters by September 2004 and lived in Maryland; Washington, DC; northern Virginia; and eastern West Virginia. Immediate neurologic symptoms postinjury were used to identify mild TBI. Adjusted prevalence ratios and 95% confidence intervals were computed by using Poisson regression. About 12% of 2,235 respondents reported a history consistent with mild TBI, and 11% screened positive for PTSD. Mild TBI history was common among veterans injured by bullets/shrapnel, blasts, motor vehicle crashes, air/water transport, and falls. Factors associated with PTSD included reporting multiple injury mechanisms (prevalence ratio = 3.71 for three or more mechanisms, 95% confidence interval: 2.23, 6.19) and combat mild TBI (prevalence ratio = 2.37, 95% confidence interval: 1.72, 3.28). The strongest factor associated with postconcussive symptoms was PTSD, even after overlapping symptoms were removed from the PTSD score (prevalence ratio = 3.79, 95% confidence interval: 2.57, 5.59). PMID- 18424431 TI - Maturation of hepatitis C virus core protein by signal peptide peptidase is required for virus production. AB - Complete maturation of hepatitis C virus (HCV) core protein requires coordinate cleavage by signal peptidase and an intramembrane protease, signal peptide peptidase. We show that reducing the intracellular levels of signal peptide peptidase lowers the titer of infectious virus released from cells, indicating that it plays an important role in virus production. Proteolysis by the enzyme at a signal peptide between core and the E1 glycoprotein is needed to permit targeting of core to lipid droplets. From mutagenesis studies, introducing mutations into the core-E1 signal peptide delayed the appearance of signal peptide peptidase-processed core until between 48 and 72 h after the beginning of the infectious cycle. Accumulation of mature core at these times coincided with its localization to lipid droplets and a rise in titer of infectious HCV. Therefore, processing of core by signal peptide peptidase is a critical event in the virus life cycle. To study the stage in virus production that may be blocked by interfering with intramembrane cleavage of core, we examined the distribution of viral RNA in cells harboring the core-E1 signal peptide mutant. Results revealed that colocalization of core with HCV RNA required processing of the protein by signal peptide peptidase. Our findings provide new insights into the sequence requirements for proteolysis by signal peptide peptidase. Moreover, they offer compelling evidence for a function for an intramembrane protease to facilitate the association of core with viral genomes, thereby creating putative sites for assembly of nascent virus particles. PMID- 18424430 TI - Expanded roles for Chk1 in genome maintenance. AB - Chk1 is a conserved kinase that imposes cell cycle delays in response to impediments to DNA replication. Recent experiments have further defined effects of Chk1 on the activity of mammalian origins of DNA replication and progression of replication forks. Moreover, Chk1 now appears to help defend genomic integrity through effects on several other pathways, including Fanconi anemia proteins, the mitotic spindle, and transcription of cell cycle-related genes. These findings can account for the requirement for Chk1 in normal proliferating cells of the early embryo and suggest the potential for diverse effects of Chk1 inhibition in cancer therapy. PMID- 18424432 TI - Nitric oxide production from nitrite occurs primarily in tissues not in the blood: critical role of xanthine oxidase and aldehyde oxidase. AB - Recent studies have shown that nitrite is an important storage form and source of NO in biological systems. Controversy remains, however, regarding whether NO formation from nitrite occurs primarily in tissues or in blood. Questions also remain regarding the mechanism, magnitude, and contributions of several alternative pathways of nitrite-dependent NO generation in biological systems. To characterize the mechanism and magnitude of NO generation from nitrite, electron paramagnetic resonance spectroscopy, chemiluminescence NO analyzer, and immunoassays of cGMP formation were performed. The addition of nitrite triggered a large amount of NO generation in tissues such as heart and liver, but only trace NO production in blood. Carbon monoxide increased NO release from blood, suggesting that hemoglobin acts to scavenge NO not to generate it. Administration of the xanthine oxidase (XO) inhibitor oxypurinol or aldehyde oxidase (AO) inhibitor raloxifene significantly decreased NO generation from nitrite in heart or liver. NO formation rates increased dramatically with decreasing pH or with decreased oxygen tension. Isolated enzyme studies further confirm that XO and AO, but not hemoglobin, are critical nitrite reductases. Overall, NO generation from nitrite mainly occurs in tissues not in the blood, with XO and AO playing critical roles in nitrite reduction, and this process is regulated by pH, oxygen tension, nitrite, and reducing substrate concentrations. PMID- 18424433 TI - The COP9/signalosome increases the efficiency of von Hippel-Lindau protein ubiquitin ligase-mediated hypoxia-inducible factor-alpha ubiquitination. AB - Oxygen-dependent ubiquitination of the alpha-subunit of hypoxia-inducible factor (HIF-alpha) by the (von Hippel-Lindau protein)-Elongin B/C-Cullin2-Rbx1 (VBC Cul2) ubiquitin ligase, a member of the cullin-RING ubiquitin ligases (CRLs), plays a central role in controlling oxygen metabolism. Nedd8 conjugation of cullins enhances the ligase activity of CRLs, and the COP9/signalosome (CSN) enhances the degradation of several CRL substrates, although it removes Nedd8 from cullins. Here we demonstrate that CSN increased the efficiency of the VBC Cul2 complex for recognizing and ubiquitinating substrates by facilitating the dissociation of ubiquitinated substrates from the pVHL subunit of the complex. Moreover CSN enhanced HIF-1alpha degradation by promoting the dissociation of HIF 1alpha from pVHL in cells. The length of the polyubiquitin chain conjugated to the substrate appeared to be involved in CSN-mediated dissociation of the substrate from pVHL. In contrast to other mechanisms underlying CSN-mediated activation of CRLs, the dissociation of ubiquitinated substrates from pVHL did not require the deneddylation activity of CSN, implying that CSN enhances degradation of CRL substrates by multiple mechanisms. PMID- 18424434 TI - Antitopes define preferential proteasomal cleavage site usage. AB - Protein degradation by proteasomes is a major source of peptides presented by major histocompatibility v complex class I proteins. Importantly, interferon gamma-induced immunoproteasomes in many cases strongly enhance the generation of antigenic peptides both in vitro and in vivo. Whether this is due to enhanced substrate turnover or to a change in proteasomal cleavage specificity is, however, largely unresolved. To overcome the problems of peptide quantification inherent to mass spectrometry, we introduced the "antitope" as substrate-specific internal standard. The antitope is a non-functional peptide that is generated by proteasomal cleavage within the epitope, resulting in partial overlaps with the functional epitope. Using antitopes as internal standards we demonstrate that the observed enhanced immunoproteasome-dependent presentation of the bacterial listeriolysin O T-cell epitope LLO(296-304) is indeed due to altered cleavage preferences. This method is also applicable to other major histocompatibility class I epitopes as is shown for two potential epitopes derived from Coxsackievirus. PMID- 18424435 TI - Regulation of the noradrenaline neurotransmitter phenotype by the transcription factor AP-2beta. AB - AP-2 family transcription factors are essential for development and morphogenesis of diverse tissues and organs, but their precise roles in specification of neural crest stem cell (NCSC)-derived cell types have not been determined. Among three members known to be expressed in the NCSC (i.e. AP-2alpha, AP-2beta, and AP 2gamma), we found that only AP-2beta is predominantly expressed in the sympathetic ganglia of developing mouse embryos, supporting its role in sympathetic development. Indeed, AP-2beta null mice expressed significantly reduced levels of both noradrenaline (NA) and NA-synthesizing dopamine beta hydroxylase in the peripheral nervous system. Strikingly, we also found that NA neuron development was significantly compromised in the locus coeruleus as well. Pharmacological treatment with an NA intermediate during pregnancy significantly rescues the neonatal lethality of AP-2beta(-/-) mice, indicating that NA deficiency is one of the main causes for lethality found in AP-2beta(-/-) mice. We also showed that forced expression of AP-2beta, but not other AP-2 factors, in NCSC favors their differentiation into NA neurons. In summary, we propose that AP 2beta plays critical and distinctive roles in the NA phenotype specification in both the peripheral and central nervous system during development. PMID- 18424436 TI - The influence of iron on Pseudomonas aeruginosa physiology: a regulatory link between iron and quorum sensing. AB - In iron-replete environments, the Pseudomonas aeruginosa Fur (ferric uptake regulator) protein represses expression of two small regulatory RNAs encoded by prrF1 and prrF2. Here we describe the effects of iron and PrrF regulation on P. aeruginosa physiology. We show that PrrF represses genes encoding enzymes for the degradation of anthranilate (i.e. antABC), a precursor of the Pseudomonas quinolone signal (PQS). Under iron-limiting conditions, PQS production was greatly decreased in a DeltaprrF1,2 mutant as compared with wild type. The addition of anthranilate to the growth medium restored PQS production to the DeltaprrF1,2 mutant, indicating that its defect in PQS production is a consequence of anthranilate degradation. PA2511 was shown to encode an anthranilate-dependent activator of the ant genes and was subsequently renamed antR. AntR was not required for regulation of antA by PrrF but was required for optimal iron activation of antA. Furthermore, iron was capable of activating both antA and antR in a DeltaprrF1,2 mutant, indicating the presence of two distinct yet overlapping pathways for iron activation of antA (AntR-dependent and PrrF dependent). Additionally, several quorum-sensing regulators, including PqsR, influenced antA expression, demonstrating that regulation of anthranilate metabolism is intimately woven into the quorum-sensing network of P. aeruginosa. Overall, our data illustrate the extensive control that both iron regulation and quorum sensing exercise in basic cellular physiology, underlining how intermediary metabolism can affect the regulation of virulence factors in P. aeruginosa. PMID- 18424437 TI - Glycogen synthase kinase (GSK) 3beta directly phosphorylates Serine 212 in the regulatory loop and inhibits microtubule affinity-regulating kinase (MARK) 2. AB - MARK/Par-1, a kinase family with diverse functions particularly in inducing cell polarity, can phosphorylate microtubule-associated proteins in their repeat domain and cause their detachment from microtubules, and thereby microtubule destabilization. Because of its role in abnormal phosphorylation of the Tau protein in Alzheimer disease, we searched for regulatory kinases. MARK family kinases can be activated by phosphorylation of a conserved threonine (Thr-208 in MARK2), and inactivated by phosphorylation of a serine (Ser-212), both in the activation loop of the catalytic domain. Activation is achieved by the kinases MARKK/TAO1 or LKB1, although the inactivating kinase was unknown. We show here that GSK3beta serves the role of the inhibitory kinase. Because GSK3beta can also phosphorylate Tau at sites outside the repeat domain, the activation of GSK3beta, and concomitant inactivation of MARK can shift the pattern of pathological phosphorylation of Tau protein in Alzheimer disease. PMID- 18424438 TI - NF-kappaB-dependent transcriptional activation in lung carcinoma cells by farnesol involves p65/RelA(Ser276) phosphorylation via the MEK-MSK1 signaling pathway. AB - In this study, we demonstrate that treatment of human lung adenocarcinoma H460 cells with farnesol induces the expression of a number of immune response and inflammatory genes, including IL-6, CXCL3, IL-1alpha, and COX-2. This response was dependent on the activation of the NF-kappaB signaling pathway. Farnesol treatment reduces the level of IkappaBalpha and induces translocation of p65/RelA to the nucleus, its phosphorylation at Ser(276), and transactivation of NF-kappaB dependent transcription. Moreover, overexpression of IkappaBalpha or treatment with the NF-kappaB inhibitor caffeic acid phenethyl ester greatly diminishes the induction of inflammatory gene expression by farnesol. We provide evidence indicating that the farnesol-induced phosphorylation of p65/RelA at Ser(276) is important for optimal transcriptional activity of NF-kappaB. The MEK1/2 inhibitor U0126 and knockdown of MEK1/2 expression with small interfering RNAs effectively blocked the phosphorylation of p65/RelA(Ser(276)) but not that of Ser(536), suggesting that this phosphorylation is dependent on the activation of the MEK1/2 ERK1/2 pathway. We further show that inhibition of MSK1, a kinase acting downstream of MEK1/2-ERK1/2, by H89 or knockdown of MSK1 expression also inhibited phosphorylation of p65/RelA(Ser(276)), suggesting that this phosphorylation is dependent on MSK1. Knockdown of MEK1/2 or MSK1 expression inhibits farnesol-induced expression of CXCL3, IL-1alpha, and COX-2 mRNA. Our results indicate that the induction of inflammatory genes by farnesol is mediated by the activation of the NF-kappaB pathway and involves MEK1/2-ERK1/2-MSK1 dependent phosphorylation of p65/RelA(Ser(276)). The activation of the NF-kappaB pathway by farnesol might be part of a prosurvival response during farnesol induced ER stress. PMID- 18424439 TI - Foxo3 is essential for the regulation of ataxia telangiectasia mutated and oxidative stress-mediated homeostasis of hematopoietic stem cells. AB - Unchecked accumulation of reactive oxygen species (ROS) compromises maintenance of hematopoietic stem cells. Regulation of ROS by the tumor suppressor protein ataxia telangiectasia mutated (ATM) is critical for preserving the hematopoietic stem cell pool. In this study we demonstrate that the Foxo3 member of the Forkhead Box O (FoxO) family of transcription factors is essential for normal ATM expression. In addition, we show that loss of Foxo3 leads to defects in hematopoietic stem cells, and these defects result from an overaccumulation of ROS. Foxo3 suppression of ROS in hematopoietic stem cells is mediated partly by regulation of ATM expression. We identify ROS-independent modulations of ATM and p16(INK4a) and ROS-mediated activation of p53/p21(CIP1/WAF1/Sdi1) tumor suppressor pathways as major contributors to Foxo3-null hematopoietic stem cells defects. Our studies demonstrate that Foxo3 represses ROS in part via regulation of ATM and that this repression is required for maintenance of the hematopoietic stem cell pool. PMID- 18424440 TI - Detailed mechanisms of the inactivation of factor VIIIa by activated protein C in the presence of its cofactors, protein S and factor V. AB - Factor VIIIa is inactivated by a combination of two mechanisms. Activation of factor VIII by thrombin results in a heterotrimeric factor VIIIa that spontaneously inactivates due to dissociation of the A2 subunit. Additionally, factor VIIIa is cleaved by the anticoagulant serine protease, activated protein C, at two cleavage sites, Arg(336) in the A1 subunit and Arg(562) in the A2 subunit. We previously characterized an engineered variant of factor VIII which contains a disulfide bond between the A2 and the A3 subunits that prevents the spontaneous dissociation of the A2 subunit following thrombin activation. Thus, in the absence of activated protein C, this variant has stable activity following activation by thrombin. To isolate the effects of the individual activated protein C cleavage sites on factor VIIIa, we engineered mutations of the activated protein C cleavage sites into the disulfide bond-cross-linked factor VIII variant. Arg(336) cleavage is 6-fold faster than Arg(562) cleavage, and the Arg(336) cleavage does not fully inactivate factor VIIIa when A2 subunit dissociation is blocked. Protein S enhances both cleavage rates but enhances Arg(562) cleavage more than Arg(336) cleavage. Factor V also enhances both cleavage rates when protein S is present. Factor V enhances Arg(562) cleavage more than Arg(336) cleavage as well. As a result, in the presence of both activated protein C cofactors, Arg(336) cleavage is only twice as fast as Arg(562) cleavage. Therefore, both cleavages contribute significantly to factor VIIIa inactivation. PMID- 18424441 TI - The stereochemical course of 4-hydroxy-2-nonenal metabolism by glutathione S transferases. AB - 4-Hydroxy-2-nonenal (HNE) is a toxic aldehyde generated during lipid peroxidation and has been implicated in a variety of pathological states associated with oxidative stress. Glutathione S-transferase (GST) A4-4 is recognized as one of the predominant enzymes responsible for the metabolism of HNE. However, substrate and product stereoselectivity remain to be fully explored. The results from a product formation assay indicate that hGSTA4-4 exhibits a modest preference for the biotransformation of S-HNE in the presence of both enantiomers. Liquid chromatography mass spectrometry analyses using the racemic and enantioisomeric HNE substrates explicitly demonstrate that hGSTA4-4 conjugates glutathione to both HNE enantiomers in a completely stereoselective manner that is not maintained in the spontaneous reaction. Compared with other hGST isoforms, hGSTA4 4 shows the highest degree of stereoselectivity. NMR experiments in combination with simulated annealing structure determinations enabled the determination of stereochemical configurations for the GSHNE diastereomers and are consistent with an hGSTA4-4-catalyzed nucleophilic attack that produces only the S-configuration at the site of conjugation, regardless of substrate chirality. In total these results indicate that hGSTA4-4 exhibits an intriguing combination of low substrate stereoselectivity with strict product stereoselectivity. This behavior allows for the detoxification of both HNE enantiomers while generating only a select set of GSHNE diastereomers with potential stereochemical implications concerning their effects and fates in biological tissues. PMID- 18424442 TI - Comprehensive transcriptional analysis of the oxidative response in yeast. AB - The oxidative stress response in Saccharomyces cerevisiae has been analyzed by parallel determination of mRNA levels and transcription rates for the entire genome. A mathematical algorithm has been adapted for a dynamic situation such as the response to stress, to calculate theoretical mRNA decay rates from the experimental data. Yeast genes have been grouped into 25 clusters according to mRNA level and transcription rate kinetics, and average mRNA decay rates have been calculated for each cluster. In most of the genes, changes in one or both experimentally determined parameters occur during the stress response. 24% of the genes are transcriptionally induced without an increase in mRNA levels. The lack of parallelism between the evolution of the mRNA amount and transcription rate predicts changes in mRNA stability during stress. Genes for ribosomal proteins and rRNA processing enzymes are abundant among those whose mRNAs are predicted to destabilize. The number of genes whose mRNAs are predicted to stabilize is lower, although some protein folding or proteasomal genes are among the latter. We have confirmed the mathematical predictions for several genes pertaining to different clusters by experimentally determining mRNA decay rates using the regulatable tetO promoter in transcriptional expression conditions not affected by the oxidative stress. This study indicates that the oxidative stress response in yeast cells is not only conditioned by gene transcription but also by the mRNA decay dynamics and that this complex response may be particularly relevant to explain the temporary down-regulation of protein synthesis occurring during stress. PMID- 18424443 TI - Testosterone-inducible regulator is a kinase that drives steroid sensing and metabolism in Comamonas testosteroni. AB - The mechanism of gene regulation by steroids in bacteria is still a mystery. We use steroid-inducible 3alpha-hydroxysteroid dehydrogenase/carbonyl reductase (3alpha-HSD/CR) as a reporter system to study steroid signaling in Comamonas testosteroni. In previous investigations we cloned and characterized the 3alpha HSD/CR-encoding gene, hsdA. In addition, we identified two negative regulator genes (repA and repB) in the vicinity of hsdA, the protein products which repress hsdA expression on the level of transcription and translation, respectively. Recently, a positive regulator of hsdA expression, TeiR (testosterone-inducible regulator), was found by transposon mutagenesis, but the mode of its action remained obscure. In the present work we produced a TeiR-green fluorescent fusion protein and showed that TeiR is a membrane protein with asymmetrical localization at one of the cell poles of C. testosteroni. Knock-out mutants of the teiR gene revealed that TeiR provides swimming and twitching motility of C. testosteroni to the steroid substrate source. TeiR also mediated an induced expression of 3alpha HSD/CR which was paralleled by an enhanced catabolism of testosterone. We also found that TeiR responds to a variety of different steroids other than testosterone. Biochemical analysis with several deletion mutants of the teiR gene revealed TeiR to consist of three different functional domains, an N-terminal domain important for membrane association, a central steroid binding site, and a C-terminal part mediating TeiR function. Finally, we could demonstrate that TeiR works as a kinase in the steroid signaling chain in C. testosteroni. Overall, we provide evidence that TeiR mediates steroid sensing and metabolism in C. testosteroni via its steroid binding and kinase activity. PMID- 18424444 TI - Overexpression of mitochondrial methionine sulfoxide reductase B2 protects leukemia cells from oxidative stress-induced cell death and protein damage. AB - According to the mitochondrial theory of aging, mitochondrial dysfunction increases intracellular reactive oxidative species production, leading to the oxidation of macromolecules and ultimately to cell death. In this study, we investigated the role of the mitochondrial methionine sulfoxide reductase B2 in the protection against oxidative stress. We report, for the first time, that overexpression of methionine sulfoxide reductase B2 in mitochondria of acute T lymphoblastic leukemia MOLT-4 cell line, in which methionine sulfoxide reductase A is missing, markedly protects against hydrogen peroxide-induced oxidative stress by scavenging reactive oxygen species. The addition of hydrogen peroxide provoked a time-gradual increase of intracellular reactive oxygen species, leading to a loss in mitochondrial membrane potential and to protein carbonyl accumulation, whereas in methionine sulfoxide reductase B2-overexpressing cells, intracellular reactive oxygen species and protein oxidation remained low with the mitochondrial membrane potential highly maintained. Moreover, in these cells, delayed apoptosis was shown by a decrease in the cleavage of the apoptotic marker poly(ADP-ribose) polymerase-1 and by the lower percentage of Annexin-V-positive cells in the late and early apoptotic stages. We also provide evidence for the protective mechanism of methionine sulfoxide reductase B2 against protein oxidative damages. Our results emphasize that upon oxidative stress, the overexpression of methionine sulfoxide reductase B2 leads to the preservation of mitochondrial integrity by decreasing the intracellular reactive oxygen species build-up through its scavenging role, hence contributing to cell survival and protein maintenance. PMID- 18424445 TI - Clinical applications of musculoskeletal tissue engineering. AB - BACKGROUND: Current surgical techniques for the repair of the musculoskeletal system can be often limited by the availability, quality and quantity of materials, such as grafts to effect repair. This has led to the exploration and development of novel methods of intervention based on tissue engineering and regenerative medicine. SOURCE OF DATA: This review summarizes the successes and investigations which are happening to date in the field of musculoskeletal tissue engineering. This is based on an extensive literature search and through basic research being performed by the authors. AREAS OF AGREEMENT: Due to the constraints surrounding certain surgical techniques and restrictions on their use, novel procedures are required for the repair and regeneration of damaged tissues. AREAS OF CONTROVERSY: The choice of cell type has caused much debate within the tissue-engineering field. However it is widely accepted that currently only autologous primary/adult stem cells are fit for transplantation, until such times that optimized differentiation and selection protocols exist for embryonic stem cells. GROWING POINTS: The current results of the clinical cases utilizing tissue engineered constructs for bone and cartilage repair provide insights for improvement of these techniques thus allowing treatments to become increasingly viable. AREAS TIMELY FOR DEVELOPING RESEARCH: There is a need to better understand the integration of scaffolds and cell populations into the target tissue. This should provide vital information influencing scaffold manufacturing procedures and cell selection. PMID- 18424446 TI - Multiple source surveillance incidence and aetiology of out-of-hospital sudden cardiac death in a rural population in the West of Ireland. AB - AIMS: There is a paucity of published data on prospectively identified rates of out-of-hospital sudden cardiac death (SCD). We sought to determine the incidence, survival and aetiology of out-of-hospital SCD in the West of Ireland for the year 2005. METHODS AND RESULTS: Data from emergency room resuscitation records were collected throughout the year from all hospitals in the West of Ireland and recorded according to pre-specified criteria. Hospital records of survivors were analysed. Simultaneously, autopsy reports from all pathology laboratories in the region were systematically reviewed and cases of SCD identified. Cardiac arrest associated with non-cardiac pathology was excluded. The population base was 414,277. There were 212 recorded cases of out-of-hospital SCD; 160 (75.5%) were male and the mean age was 63.3 years. The incidence rate was 51.2/100,000/year. The most common aetiology was coronary artery disease (161 cases; 75.9%). The majority of cases occurred in the home (152, 71.7%). Thirteen (6.1%) patients survived to admission of whom eight (3.8%) were alive at discharge. All survivors had ventricular fibrillation as the presenting rhythm. CONCLUSION: The burden of SCD in the West of Ireland is considerable. The vast majority of cases occur in the home. Survival rates in this rural population cohort remain low. PMID- 18424447 TI - Molecular dissection of ALS-associated toxicity of SOD1 in transgenic mice using an exon-fusion approach. AB - Mutations in Cu,Zn superoxide dismutase (SOD1) are associated with amyotrophic lateral sclerosis (ALS). Among more than 100 ALS-associated SOD1 mutations, premature termination codon (PTC) mutations exclusively occur in exon 5, the last exon of SOD1. The molecular basis of ALS-associated toxicity of the mutant SOD1 is not fully understood. Here, we show that nonsense-mediated mRNA decay (NMD) underlies clearance of mutant mRNA with a PTC in the non-terminal exons. To further define the crucial ALS-associated SOD1 fragments, we designed and tested an exon-fusion approach using an artificial transgene SOD1(T116X) that harbors a PTC in exon 4. We found that the SOD1(T116X) transgene with a fused exon could escape NMD in cellular models. We generated a transgenic mouse model that overexpresses SOD1(T116X). This mouse model developed ALS-like phenotype and pathology. Thus, our data have demonstrated that a 'mini-SOD1' of only 115 amino acids is sufficient to cause ALS. This is the smallest ALS-causing SOD1 molecule currently defined. This proof of principle result suggests that the exon-fusion approach may have potential not only to further define a shorter ALS-associated SOD1 fragment, thus providing a molecular target for designing rational therapy, but also to dissect toxicities of other proteins encoded by genes of multiple exons through a 'gain of function' mechanism. PMID- 18424448 TI - Variants in the estrogen receptor alpha gene and its mRNA contribute to risk for schizophrenia. AB - Estrogen modifies human emotion and cognition and impacts symptoms of schizophrenia. We hypothesized that the variation in the estrogen receptor alpha (ESR1) gene and cortical ESR1 mRNA is associated with schizophrenia. In a small case-control genetic association analysis of postmortem brain tissue, genotype CC (rs2234693) and haplotypes containing the C allele of a single-nucleotide polymorphism (SNP) in intron1 (PvuII) were more frequent in African American schizophrenics (P = 0.01-0.001). In a follow-up family-based association analysis, we found overtransmission of PvuII allele C and a PvuII C-containing haplotype (P = 0.01-0.03) to African American and Caucasian patients with schizophrenia. Schizophrenics with the 'at risk' PvuII genotype had lower ESR1 mRNA levels in the frontal cortex. Eighteen ESR1 splice variants and decreased frequencies of the wild-type ESR1 mRNA were detected in schizophrenia. In one patient, a unique ESR1 transcript with a genomic insert encoding a premature stop codon and a truncated ESR1 protein lacking most of the estrogen binding domain was the only transcript detected. Using a luciferase assay, we found that mRNA encoding a truncated ESR1 significantly attenuates gene expression at estrogen response elements demonstrating a dominant negative function. An intron 6 SNP [rs2273207(G)] was associated with an ESR1 splice variant missing exon seven. The T allele of another intron 6 SNP was part of a 3' haplotype less common in schizophrenia [rs2273206(T), rs2273207(G), rs2228480(G)]. Thus, the variation in the ESR1 gene is associated with schizophrenia and the mechanism of this association may involve alternative gene regulation and transcript processing. PMID- 18424449 TI - Iron-dependent regulation of frataxin expression: implications for treatment of Friedreich ataxia. AB - Friedreich ataxia (FA) is a progressive neurodegenerative disease caused by expansion of a trinucleotide repeat within the first intron of the gene that encodes frataxin. In our study, we investigated the regulation of frataxin expression by iron and demonstrated that frataxin mRNA levels decrease significantly in multiple human cell lines treated with the iron chelator, desferal (DFO). In addition, frataxin mRNA and protein levels decrease in fibroblast and lymphoblast cells derived from both normal controls and from patients with FA when treated with DFO. Lymphoblasts and fibroblasts of FA patients have evidence of cytosolic iron depletion, as indicated by increased levels of iron regulatory protein 2 (IRP2) and/or increased IRE-binding activity of IRP1. We postulate that this inferred cytosolic iron depletion occurs as frataxin-deficient cells overload their mitochondria with iron, a downstream regulatory effect that has been observed previously when mitochondrial iron sulfur cluster assembly is disrupted. The mitochondrial iron overload and presumed cytosolic iron depletion potentially further compromise function in frataxin-deficient cells by decreasing frataxin expression. Thus, our results imply that therapeutic efforts should focus on an approach that combines iron removal from mitochondria with a treatment that increases cytosolic iron levels to maximize residual frataxin expression in FA patients. PMID- 18424450 TI - Sphingosine kinase 1/S1P receptor signaling axis controls glial proliferation in mice with Sandhoff disease. AB - Sphingosine-1-phosphate (S1P) is a lipid-signaling molecule produced by sphingosine kinase in response to a wide number of stimuli. By acting through a family of widely expressed G protein-coupled receptors, S1P regulates diverse physiological processes. Here we examined the role of S1P signaling in neurodegeneration using a mouse model of Sandhoff disease, a prototypical neuronopathic lysosomal storage disorder. When sphingosine kinase 1 (Sphk1) was deleted in Sandhoff disease mice, a milder disease course occurred, with decreased proliferation of glial cells and less-pronounced astrogliosis. A similar result of milder disease course and reduced astroglial proliferation was obtained by deletion of the gene for the S1P(3) receptor, a G protein-coupled receptor enriched in astrocytes. Our studies demonstrate a functional role of S1P synthesis and receptor expression in astrocyte proliferation leading to astrogliosis during the terminal stages of neurodegeneration in Sandhoff disease mice. Because astrocyte responses are involved in many types of neurodegeneration, the Sphk1/S1P receptor signaling axis may be generally important during the pathogenesis of neurodegenerative diseases. PMID- 18424451 TI - Uncoupling of chondrocyte differentiation and perichondrial mineralization underlies the skeletal dysplasia in tricho-rhino-phalangeal syndrome. AB - Tricho-rhino-phalangeal syndrome (TRPS) is an autosomal dominant craniofacial and skeletal dysplasia that is caused by mutations involving the TRPS1 gene. Patients with TRPS have short stature, hip abnormalities, cone-shaped epiphyses and premature closure of growth plates reflecting defects in endochondral ossification. The TRPS1 gene encodes for the transcription factor TRPS1 that has been demonstrated to repress transcription in vitro. To elucidate the molecular mechanisms underlying skeletal abnormalities in TRPS, we analyzed Trps1 mutant mice (Trps1DeltaGT mice). Analyses of growth plates demonstrated delayed chondrocyte differentiation and accelerated mineralization of perichondrium in Trps1 mutant mice. These abnormalities were accompanied by increased Runx2 and Ihh expression and increased Indian hedgehog signaling. We demonstrated that Trps1 physically interacts with Runx2 and represses Runx2-mediated trans activation. Importantly, generation of Trps1(DeltaGT/+);Runx2(+/-) double heterozygous mice rescued the opposite growth plate phenotypes of single mutants, demonstrating the genetic interaction between Trps1 and Runx2 transcription factors. Collectively, these data suggest that skeletal dysplasia in TRPS is caused by dysregulation of chondrocyte and perichondrium development partially due to loss of Trps1 repression of Runx2. PMID- 18424452 TI - A nonsense mutation in the LIMP-2 gene associated with progressive myoclonic epilepsy and nephrotic syndrome. AB - The main clinical features of two siblings from a consanguineous marriage were progressive myoclonic epilepsy without intellectual impairment and a nephrotic syndrome with a strong accumulation of C1q in capillary loops and mesangium of kidney. The biochemical analysis of one of the patients revealed a normal beta glucocerebrosidase activity in leukocytes, but a severe enzymatic deficiency in cultured skin fibroblasts. This deficiency suggested a defect in the intracellular sorting pathway of this enzyme. The sequence analysis of the gene encoding LIMP-2 (SCARB2), the sorting receptor for beta-glucocerebrosidase, confirmed this hypothesis. A homozygous nonsense mutation in codon 178 of SCARB2 was found in the patient, whereas her healthy parents were heterozygous for the mutation. Besides lacking immunodetectable LIMP-2, patient fibroblasts also had decreased amounts of beta-glucocerebrosidase, which was mainly located in the endoplasmic reticulum, as assessed by its sensitivity to Endo H. This is the first report of a mutation in the SCARB2 gene associated with a human disease, which, contrary to earlier proposals, shares no features with Charcot-Marie-Tooth disease both at the clinical and neurophysiological levels. PMID- 18424453 TI - Interaction between a novel TGFB1 haplotype and CFTR genotype is associated with improved lung function in cystic fibrosis. AB - Cystic fibrosis (CF), the most common lethal single gene disorder in Caucasians, is due to mutations in the CFTR gene. Twin and sibling analysis indicates that modifier genes, rather than allelic variation in CFTR, are responsible for most of the variability in severity of lung disease, the major cause of mortality in CF patients. We used a family-based approach to test for association between lung function and two functional SNPs (rs1800469, '-509' and rs1982073, 'codon 10') in the 5' region of transforming growth factor-beta1 (TGFB1), a putative CF modifier gene. Quantitative transmission disequilibrium testing of 472 CF patient-parent parent trios revealed that both TGFB1 SNPs showed significant transmission distortion when patients were stratified by CFTR genotype. Although lung function and nutritional status are correlated in CF patients, there was no evidence of association between the TGFB1 SNPs and variation in nutritional status. Additional tagging SNPs (rs8179181, rs2278422, rs8110090, rs4803455 and rs1982072) that capture most of the diversity in TGFB1 were also typed but none showed association with variation in lung function. However, a haplotype composed of the -509 C and codon 10 T alleles along with the C allele of the 3' SNP rs8179181 was highly associated with increased lung function in patients grouped by CFTR genotype. These results demonstrate that TGFB1 is a modifier of CF lung disease and reveal a previously unrecognized beneficial effect of TGFB1 variants upon the pulmonary phenotype. PMID- 18424455 TI - Ethical issues in medico-legal exposures. AB - The Medical Exposure Directive (MED) 97/43/Euratom defines medico-legal procedures as 'procedures performed for insurance or legal purposes without a medical indication'. The term 'medico-legal exposures' covers a wide range of possible types of exposures, very different in nature, for which the only feature in common is the fact that the main reason for performing them does not relate directly to the health of the individual being exposed to ionising radiation. The key issue in medico-legal exposures is justification. Balancing the advantages and disadvantages of such exposures is complex because not only can these be difficult to quantify and hence compare, but often the advantage may be to society whereas the disadvantage is usually to an individual. This adds an additional layer of ethical complexity to the problem and one, which requires input from a number of sources beyond the established radiation protection community. Because medico-legal exposures are considered to be medical exposures, they are not subject to dose limits. In medico-legal exposures where the benefit is not necessarily to the individual undergoing the exposure, the question must be asked as to whether or not this is an appropriate framework within which to conduct such exposures. This paper looks at the current situation in Europe, highlighting some of the particular problems that have arisen, and tries to identify the areas, which require further clarification and guidance. PMID- 18424454 TI - ABCB1 (MDR1) genetic variants are associated with methadone doses required for effective treatment of heroin dependence. AB - Methadone is a mu-opioid receptor agonist used for treating opiate dependence. The range of effective methadone doses is broad. Part of the large inter individual variability in efficacy may be accounted for by genetic factors. Methadone is a substrate of the transporter P-glycoprotein (P-gp) 170 that is encoded by the ABCB1 (MDR1) gene. Thus, P-gp variants may play a role in methadone absorption and distribution. We assessed the association between ABCB1 polymorphisms and methadone dose requirements in 98 methadone-maintained patients. The stabilizing methadone doses were normally distributed with a mean and median dose of 160 mg/day (range 30-280 mg/day). Statistical analysis showed significant difference in genotype frequencies between the 'higher' (>150 mg/day) and 'lower' (< or =150 mg/day) methadone dose groups for single nucleotide polymorphism (SNP) 1236C>T (rs1128503) (experiment-wise P = 0.0325). Furthermore, individuals bearing the 3-locus genotype pattern TT-TT-TT (rs1045642, rs2032582 and rs1128503) have an approximately 5-fold chance of requiring the 'higher' methadone dose, while individuals heterozygous for these three SNPs have an approximately 3-fold chance of stabilizing at the 'lower' methadone dose (point wise P-value = 0.026). These data suggest that specific ABCB1 variants may have clinical relevance by influencing the methadone dose required to prevent withdrawal symptoms and relapse in this population. PMID- 18424456 TI - New transvenous implantable cardioverter defibrillator configuration for use after tricuspid valve surgery. AB - A 57-year-old patient with a tricuspid valve (TV) prosthesis underwent successful atrioventricular pacing and internal defibrillation via a totally transvenous approach without crossing the TV. Ventricular pacing and sensing were obtained with a bipolar lead in the lateral cardiac vein. Internal defibrillation was obtained with a coil lead in the middle cardiac vein and an 'active can' pulse generator in the left infraclavicular region. PMID- 18424457 TI - New method for cardiac resynchronization therapy: transapical endocardial lead implantation for left ventricular free wall pacing. AB - Coronary sinus lead placement for transvenous left ventricular (LV) pacing in cardiac resynchronization therapy (CRT) has a significant failure rate at implant and a significant dislocation rate during follow-up. For these patients, epicardial pacing lead implantation is the most frequently used alternative. The aim of this case report is to describe a fundamentally new approach for the endocardial LV lead implantation. An epicardial lead implantation was planned, but after thoracotomy, extensive pericardial adhesions were found. An active fixation lead was placed into the LV cavity using the standard Seldinger technique through the LV apex. After an uneventful post-operative period at the 3 and 6-month follow-up visits, the patient had effective CRT with unchanged pacing parameters. In conclusion, this is the very first report showing feasibility of transapical LV lead implantation. PMID- 18424458 TI - Fast grid layout algorithm for biological networks with sweep calculation. AB - MOTIVATION: Properly drawn biological networks are of great help in the comprehension of their characteristics. The quality of the layouts for retrieved biological networks is critical for pathway databases. However, since it is unrealistic to manually draw biological networks for every retrieval, automatic drawing algorithms are essential. Grid layout algorithms handle various biological properties such as aligning vertices having the same attributes and complicated positional constraints according to their subcellular localizations; thus, they succeed in providing biologically comprehensible layouts. However, existing grid layout algorithms are not suitable for real-time drawing, which is one of requisites for applications to pathway databases, due to their high computational cost. In addition, they do not consider edge directions and their resulting layouts lack traceability for biochemical reactions and gene regulations, which are the most important features in biological networks. RESULTS: We devise a new calculation method termed sweep calculation and reduce the time complexity of the current grid layout algorithms through its encoding and decoding processes. We conduct practical experiments by using 95 pathway models of various sizes from TRANSPATH and show that our new grid layout algorithm is much faster than existing grid layout algorithms. For the cost function, we introduce a new component that penalizes undesirable edge directions to avoid the lack of traceability in pathways due to the differences in direction between in-edges and out-edges of each vertex. AVAILABILITY: Java implementations of our layout algorithms are available in Cell Illustrator. CONTACT: masao@ims.u tokyo.ac.jp SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online. PMID- 18424459 TI - Metabolomic profiling by near-infrared spectroscopy as a tool to assess embryo viability: a novel, non-invasive method for embryo selection. AB - BACKGROUND: The morphology of an embryo has a limited predictive value for assessing viability and ongoing pregnancy, therefore new selection tools are needed to maintain success rates with single-embryo transfer (SET). In this study, we investigated if metabolomic profiling of biomarkers of embryo culture medium by near-infrared (NIR) spectroscopy has a correlation with ongoing pregnancy in SET. METHODS: A total of 333 patients scheduled for in vitro fertilization (IVF) with SET were included in the study. Embryos were selected for transfer by morphological criteria on Days 2 and 3 of in vitro culture, and left over culture media samples were analyzed by NIR spectroscopy. RESULTS: The NIR spectral analysis produced unique metabolomic profiles that correlated to an embryo's reproductive potential. Resulting relative viability scores between positive and negative pregnancy outcomes were statistically significant (P < 0.03). A logistic regression of factors correlated to pregnancy outcomes showed that maternal age, percent fragmentation and relative viability scores all demonstrated a relationship. The extent of the correlation was determined by accuracy computation, where the accuracy of assessing viable embryos on Day 3 by metabolomic profiling was 53.6% and the accuracy of the morphological selection was 38.5%. In addition, the positive predictive value of metabolomic profiling was 0.365 and the negative predictive value was 0.830. CONCLUSIONS: NIR metabolomic profiling of spent embryo culture media was able to distinguish viable embryos from non-viable embryos for reproduction. PMID- 18424460 TI - Inhibition of mineralocorticoid receptors with eplerenone alleviates short-term cyclosporin A nephrotoxicity in conscious rats. AB - BACKGROUND: Recent data indicate that aldosterone aggravates cyclosporin A (CsA) induced nephrotoxicity. We examined whether the mineralocorticoid receptor (MR) blocker eplerenone (EPL) antagonized early deterioration of renal function and blood pressure (BP) increase in CsA-treated rats. METHODS: Male Sprague-Dawley rats received CsA (15 mg/kg/day i.p.) and/or EPL (100 mg/kg/day p.o.) for 21 days. After 2 weeks, arterial, venous and urinary bladder catheters were implanted and the rats were trained to accept a restraining device allowing arterial blood sampling and direct measurement of BP and renal function. BP was measured on-line in conscious rats. RESULTS: CsA significantly increased systolic BP: 139 +/- 4 versus 134 +/- 2 mmHg, reduced body weight gain: -5 +/- 6 versus 36 +/- 7 g, glomerular filtration rate (GFR): 1.02 +/- 0.16 versus 2.64 +/- 0.27 ml/min, renal blood flow (RBF): 5.3 +/- 2.4 versus 13.5 +/- 2.1 ml/min and lithium clearance (C(Li+)): 0.16 +/- 0.04 versus 0.26 +/- 0.07 ml/min compared to controls. These changes were prevented by simultaneous EPL treatment: systolic BP, 130 +/- 4 mmHg; weight gain, 53 +/- 7 g; GFR, 1.67 +/- 0.26 ml/min; RBF, 12.3 +/- 2.1 ml/min and C(Li+), 0.27 +/- 0.03 ml/min. Analysis of kidney morphology after the CsA treatment showed hyaline vacuolization in tubules and vascular depositions in arterioles; these changes were less pronounced after combination therapy. No significant changes were seen regarding haemoglobin, haematocrit, plasma renin and vasopressin, plasma and urinary sodium, potassium, or osmolality. CONCLUSIONS: MR blockade by EPL prevented short-term alterations in GFR, RBF and hypertension associated with CsA nephrotoxicity. We conclude that the aldosterone-MR pathway contributes markedly to the renal toxicity induced by this calcineurin inhibitor. PMID- 18424461 TI - A practice-related risk score (PRS): a DOPPS-derived aggregate quality index for haemodialysis facilities. AB - BACKGROUND: The Dialysis Outcomes and Practice Patterns Study (DOPPS) database was used to develop and validate a practice-related risk score (PRS) based on modifiable practices to help facilities assess potential areas for improving patient care. METHODS: Relative risks (RRs) from a multivariable Cox mortality model, based on observational haemodialysis (HD) patient data from DOPPS I (1996 2001, seven countries), were used. The four practices were the percent of patients with Kt/V > or =1.2, haemoglobin > or =11 g/dl (110 g/l), albumin > or =4.0 g/dl (40g/l) and catheter use, and were significantly related to mortality when modelled together. DOPPS II data (2002-2004, 12 countries) were used to evaluate the relationship between PRS and mortality risk using Cox regression. RESULTS: For facilities in DOPPS I and II, changes in PRS over time were significantly correlated with changes in the standardized mortality ratio (SMR). The PRS ranged from 1.0 to 2.1. Overall, the adjusted RR of death was 1.05 per 0.1 points higher PRS (P < 0.0001). For facilities in both DOPPS I and II (N = 119), a 0.2 decrease in PRS was associated with a 0.19 decrease in SMR (P = 0.005). On average, facilities that improved PRS practices showed significantly reduced mortality over the same time frame. CONCLUSIONS: The PRS assesses modifiable HD practices that are linked to improved patient survival. Further refinements might lead to improvements in the PRS and will address regional variations in the PRS/mortality relationship. PMID- 18424462 TI - New recommendations in the treatment of Gram-positive bacteraemia in dialysis patients. PMID- 18424464 TI - Clinical research of kidney diseases V: extended analytic models. PMID- 18424465 TI - Pseudohypoaldosteronisms, report on a 10-patient series. AB - BACKGROUND: Type 1 pseudohypoaldosteronism (PHA1) is a salt-wasting syndrome caused by mineralocorticoid resistance. Autosomal recessive and dominant hereditary forms are caused by Epithelial Na Channel and Mineralocorticoid Receptor mutation respectively, while secondary PHA1 is usually associated with urological problems. METHODS: Ten patients were studied in four French pediatric units in order to characterize PHA1 spectrum in infants. Patients were selected by chart review. Genetic, clinical and biochemistry data were collected and analyzed. RESULTS: Autosomal recessive PHA1 (n = 3) was diagnosed at 6 and 7 days of life in three patients presenting with severe hyperkalaemia and weight loss. After 8 months, 3 and 5 years on follow-up, neurological development and longitudinal growth was normal with high sodium supplementation. Autosomal dominant PHA1 (n = 4) was revealed at 15, 19, 22 and 30 days of life because of failure to thrive. At 8 months, 3 and 21 years of age, longitudinal growth was normal in three patients who were given salt supplementation; no significant catch-up growth was obtained in the last patient at 20 months of age. Secondary PHA1 (n = 3) was diagnosed at 11, 26 days and 5 months of life concomitantly with acute pyelonephritis in three children with either renal hypoplasia, urinary duplication or bilateral megaureter. The outcome was favourable and salt supplementation was discontinued after 3, 11 and 13 months. CONCLUSIONS: PHA1 should be suspected in case of severe hyperkalemia and weight loss in infants and need careful management. Pathogenesis of secondary PHA1 is still challenging and further studies are mandatory to highlight the link between infection, developing urinary tract and pseudohypoaldosteronism. PMID- 18424466 TI - Vasculitis and anti-thyroid medication. PMID- 18424468 TI - An update on UK rheumatology consultant workforce provision: the BSR/ARC Workforce Register 2005-07: assessing the impact of recent changes in NHS provision. AB - OBJECTIVES: To describe changes in the provision of rheumatology services, monitor the pattern of inequalities in UK rheumatology service provision since 2005, and to summarize the 3-yr impact of the new National Health Service (NHS) consultant contract and the Musculoskeletal Services Framework in England and Wales. METHODS: Questionnaires about timetable and working conditions were sent to all consultants on the BSR/ARC UK Workforce Register in January 2007, along with the personal and job-related details currently held about them on the register to update. The questionnaire included a visual analogue scale asking 'how concerned are you that your current post might be under threat' ranging from 0 'Not at all' to 100 'Extremely'. RESULTS: The response rate of the 2005 and 2007 surveys were 89 and 87%, respectively. Levels of optimal provision now exceed 70% in England and Wales, and 50% in Scotland and Northern Ireland. Levels of provision remain substantially higher in London than anywhere else. The median level of perceived job threat in the UK was 31 (interquartile range 11-61). Consultants in areas where provision is highest and a higher proportion of services are run in conjunction with Clinical Assessment and Treatment (CAT) centres report higher perceived job threat. CONCLUSIONS: Provision of rheumatology services has continued to expand over the past decade; however, inequalities persist at national and sub-national level. There is evidence of improvement in regions with the lowest provision, but there are indications of increased perceived job threat in areas with traditionally higher provision and where CAT centres have been introduced. PMID- 18424469 TI - Surveillance for rare infectious diseases: is one passive data source enough for Haemophilus influenzae? AB - BACKGROUND: The completeness of a compulsory reporting system of systemic Haemophilus influenzae infections in children in Germany is studied by means of cross-linking registry data from three sources and applying capture-recapture methods. METHODS: Cases were collected for the years 2001-05 by three national data sources: a passive administration registry (SurvNet@RKI), an active hospital surveillance system and an active laboratory surveillance system. The case definition required cultural detection of H. influenzae in blood or cerebrospinal fluid. Linkage was carried out by month and year of birth, sex, geographical region and date of disease onset. Capture-recapture models were used to estimate the incidence of invasive H. influenzae infections. RESULTS: SurvNet@RKI reported 113 H. influenzae and 38 H. influenzae type b (Hib) cases, compared to a total of 231 and 68 cases, respectively, reported by all three sources combined. Best fitting 3-source capture-recapture estimations amounts to 258 (95% confidence interval: 247-276) H. influenzae and 71 (69-74) Hib cases. SurvNet@RKI data depicted a similar decrease in annual H. influenzae cases as the capture recapture estimates but failed to detect the underlying decrease in Hib cases which was observed in the capture-recapture estimates due to a considerable annual variability of ascertainment of serotyped cases in SurvNet@RKI ranging from 14% to 69%. CONCLUSIONS: Because of small variability of ascertainment, the compulsory passive reporting system depicted trends in H. influenzae incidence, although less than half of the cases were ascertained. However, time trend in Hib cases could not be depicted, because of highly variable serotyping proportions. PMID- 18424470 TI - The oldest old in England and Wales: a descriptive analysis based on the MRC Cognitive Function and Ageing Study. AB - OBJECTIVE: to describe the characteristics and survival of the oldest old in England and Wales. DESIGN: retrospective analysis of the oldest old from a population-based cohort study. SETTING: population-based study in England and Wales: two rural and three urban sites. METHODS: two types of analyses were conducted: (i) a descriptive analysis of those individuals who were aged 90 years or more, and 100 years or more, and (ii) a survival analysis of those who reached their 90th, 95th, or 100th birthday during the study. Median survival time was calculated by the Kaplan-Meier method. Effects of socio-demographic characteristics on survival were evaluated using the Cox proportional-hazards regression model. RESULTS: in total, 958 individuals aged 90 years or more, and 24 individuals aged 100 years or more, had been interviewed at least once during the study. Twenty-seven per cent were living in residential or nursing homes. Women aged 90 years or more were more likely to be living in residential and nursing homes, be widowed, have any disability or have lower MMSE scores. The centenarians were mostly cognitively and functionally impaired. The median survival times for those reaching their 90th (n = 2,336), 95th (n = 638), or 100th birthday (n = 92) during the study were 3.7 years (95% CI: 3.5-4.0), 2.3 (2.1-2.6) and 2.1 (1.7-2.6) years for women, and 2.9 (95% CI: 2.6- 3.1), 2.0 (1.2 3.1) and 2.2 (0.5-2.3) for men, respectively. Those living in residential and nursing homes had a shorter survival when aged 90 years, with similar non significant effects for those aged 95 and 100 years. After the age of 100 years, the high mortality rate and small sample size limited the ability to detect any differences between the different groups. CONCLUSION: even at the very oldest ages, the majority live in non-institutionalised settings. Among the oldest old, women were frailer than men. Being male and living in residential nursing homes shortened survival in those aged 90 years or more. PMID- 18424471 TI - Pollination ecology and breeding systems of five Gesneria species from Puerto Rico. AB - BACKGROUND AND AIMS: The genus Gesneria diversified in the Greater Antilles giving rise to various floral designs corresponding to different pollination syndromes. The goal of this study was to characterize the pollination and breeding systems of five Puerto Rican Gesneria species. METHODS: The study was conducted in Arecibo and El Yunke National Forest, Puerto Rico, between 2003 and 2007. Floral visitors were documented by human observers and video cameras. Floral longevity and nectar production were recorded for the five study species. Tests for self-compatibility and autonomous selfing were conducted through hand pollination and bagging experiments. KEY RESULTS: Floral phenology and nectar production schedules agree with nocturnal (in bell-shaped flowered G. pedunculosa and G. viridiflora subsp. sintenisii) or diurnal pollination syndromes (in tubular-flowered G. citrina, G. cuneifolia and G. reticulata). Nectar concentration is consistently low (8-13 %) across species. Gesneria citrina and G. cuneifolia are exclusively pollinated by hummingbirds, while Gesneria reticulata relies mostly on autonomous self-pollination, despite having classic ornithophilous flowers. A variety of floral visitors was recorded for the two species with bell-shaped flowers; however, not all visitors have the ability to transfer pollen. Bats are the primary pollinators of G. pedunculosa, with bananaquits probably acting as secondary pollinators. For G. viridiflora subsp. sintenisii, both bats and hummingbirds contact the flower's reproductive organs, thus, this species is considered to be a generalist despite its nocturnal floral syndrome. All species are self-compatible but only tubular-flowered Gesneria are capable of autonomous self-pollination. CONCLUSIONS: The visitation patterns described in this study fit the predicted hummingbird and bat pollination syndromes and support both specialization and generalization of pollination systems in Puerto Rican Gesneria. Specialization is associated with low pollinator visitation, particularly by hummingbirds, which may explain the occurrence of autonomous selfing mechanisms in tubular-flowered species. PMID- 18424472 TI - Association between floral traits and rewards in Erysimum mediohispanicum (Brassicaceae). AB - BACKGROUND AND AIMS: Floral rewards may be associated with certain morphological floral traits and thus act as underlying factors promoting selection on these traits. This study investigates whether some traits that are under pollinator mediated selection (flower number, stalk height, corolla diameter, corolla tube length and corolla tube width) in the Mediterranean herb E. mediohispanicum (Brassicaceae) are associated with rewards (pollen and nectar). METHODS: During 2005 the phenotypic traits and the visitation rate of the main pollinator functional groups were quantified in 720 plants belonging to eight populations in south-east Spain, and during 2006 the same phenotypic traits and the reward production were quantified in 400 additional plants from the same populations. KEY RESULTS: A significant correlation was found between nectar production rate and corolla tube length, and between pollen production and corolla diameter. Visitation rates of large bees and butterflies were significantly higher in plants exhibiting larger flowers with longer corolla tubes. CONCLUSIONS: The association between reward production and floral traits may be a factor underlying the pattern of visitation rate displayed by some pollinators. PMID- 18424473 TI - Needlestick injuries in primary care. PMID- 18424474 TI - Price elasticity estimates for tobacco products in India. AB - The tax base of tobacco in India is heavily dependent on about 14% of tobacco users, who smoke cigarettes. Non-cigarette tobacco products accounting for 85% of the tobacco consumption contributes only 15% of the total tobacco taxes. Though taxation is an important tool to regulate consumption of tobacco, there have been no estimates of price elasticities for different tobacco products in India to date, which can guide tax policy on tobacco. This paper, for the first time in India, examines the price elasticity of demand for bidis, cigarettes and leaf tobacco at the national level using a representative cross-section of households. This study found that own-price elasticity estimates of different tobacco products in India ranged between -0.4 to -0.9, with bidis (an indigenous hand rolled smoked tobacco preparation in India) and leaf tobacco having elasticities close to unity. Cigarettes were the least price elastic of all. With some assumptions, it is shown that the tax on bidis can be increased to Rs. 100 per 1000 sticks compared with the current Rs. 14 and the tax on an average cigarette can be increased to Rs. 3.5 per stick without any fear of losing revenue. The paper argues that the current system of taxing cigarettes in India based on the presence of filters and the length of cigarettes has no justification on health grounds, and should be abolished, if reducing tobacco consumption and the consequent disease burden is one of the objectives of tobacco taxation policy. It also argues that attempts to regulate tobacco use without effecting significant tax increases on bidis may not produce desired results. PMID- 18424475 TI - Sensitivity of between-study heterogeneity in meta-analysis: proposed metrics and empirical evaluation. AB - BACKGROUND: Several approaches are available for evaluating heterogeneity in meta analysis. Sensitivity analyses are often used, but these are often implemented in various non-standardized ways. METHODS: We developed and implemented sequential and combinatorial algorithms that evaluate the change in between-study heterogeneity as one or more studies are excluded from the calculations. The algorithms exclude studies aiming to achieve either the maximum or the minimum final I(2) below a desired pre-set threshold. We applied these algorithms in databases of meta-analyses of binary outcome and >/=4 studies from Cochrane Database of Systematic Reviews (Issue 4, 2005, n = 1011) and meta-analyses of genetic associations (n = 50). Two I(2) thresholds were used (50% and 25%). RESULTS: Both algorithms have succeeded in achieving the pre-specified final I(2) thresholds. Differences in the number of excluded studies varied from 0% to 6% depending on the database and the heterogeneity threshold, while it was common to exclude different specific studies. Among meta-analyses with initial I(2) > 50%, in the large majority [19 (90.5%) and 208 (85.9%) in genetic and Cochrane meta analyses, respectively] exclusion of one or two studies sufficed to decrease I(2) < 50%. Similarly, among meta-analyses with initial I(2) > 25%, in most cases [16 (57.1%) and 382 (81.3%), respectively) exclusion of one or two studies sufficed to decrease heterogeneity even <25%. The number of excluded studies correlated modestly with initial estimated I(2) (correlation coefficients 0.52-0.68 depending on algorithm used). CONCLUSIONS: The proposed algorithms can be routinely applied in meta-analyses as standardized sensitivity analyses for heterogeneity. Caution is needed evaluating post hoc which specific studies are responsible for the heterogeneity. PMID- 18424476 TI - Clinical features of patients with multiple sclerosis from a survey in Shanghai, China. AB - OBJECTIVE: To describe clinical features of patients with multiple sclerosis (MS) in Shanghai, China. METHODS: Prevalent patients with MS were identified and investigated by a network of physicians in 11 districts of Shanghai during the period from 1 September 2004 to 31 August 2005. Admission registries of each hospital in the study area were checked systematically for patients with a diagnosis of MS, neuromyelitis optica or other demyelinating disorders. All patients with collected information were evaluated by four senior neurologists according to the McDonald criteria. RESULTS: There were 249 (146 female and 103 male) patients with a confirmed MS diagnosis, at a female-to-male ratio of 1.4. The mean age at onset of MS was 37.4 years for the 249 patients with MS and, on the prevalence day, 42.7 years. The most frequent location of clinical MS lesions in the central nervous system was the spinal cord (61%), followed by the cerebrum (55%) and optic nerves (41%). Nearly all (96%) of the patients with MS had been examined by magnetic resonance imaging, and 226 (94%) patients of those examined were suggestive of MS. No family history of MS was found in any of the patients. Most (86%) of the patients had no or mild disability on the prevalence day (31 December 2004). Almost all (96%) patients with MS had been treated with corticosteroids. CONCLUSION: Clinical features of patients with MS are described based on the information from the largest case series reported among Chinese. Comparisons and discussions are made with findings from the other populations. PMID- 18424477 TI - A case of dysphagia with clinically isolated syndrome. PMID- 18424478 TI - Magnetic resonance imaging metrics and their correlation with clinical outcomes in multiple sclerosis: a review of the literature and future perspectives. AB - Magnetic resonance imaging (MRI) has revolutionized the diagnosis and management of patients with multiple sclerosis (MS). Conventional MRI metrics are employed as primary endpoints in proof-of-concept clinical trials evaluating new drugs for MS and as secondary endpoints in definitive phase III trials. Metrics derived from non-conventional MRI techniques are now emerging and hold significant promise since they appear to be more correlated with the most disabling features of MS. However, none of these has been approved for use as a surrogate endpoint for accumulation of physical disability, which is the most important clinical endpoint of this disease. Taking into account the large numbers of patients needed, the extensive exposure to placebo, and the relatively long duration required for phase III clinical trials to show a meaningful effect on progression of disability, the need for a valid, reliable, and objective paraclinical marker of disease evolution cannot be overemphasized. This paper reviews the most up-to date data regarding MRI techniques, their relationship with central nervous system pathology, as well as with clinical endpoints, and proposes future insights into the use of MRI metrics as surrogate endpoints in clinical trials of MS. PMID- 18424479 TI - Glatiramer acetate after induction therapy with mitoxantrone in relapsing multiple sclerosis. AB - Forty relapsing multiple sclerosis patients with 1-15 gadolinium (Gd)-enhancing lesions on screening brain magnetic resonance imaging (MRI) and Expanded Disability Status Scale (EDSS) scores 0-6.5 were randomized to receive short-term induction therapy with mitoxantrone (three monthly 12 mg/m(2) infusions) followed by 12 months of daily glatiramer acetate (GA) therapy 20 mg/day subcutaneously for a total of 15 months (M-GA, n = 21) or daily GA 20 mg/day for 15 months (GA, n = 19). MRI scans were performed at months 6, 9, 12 and 15. The primary measure of outcome was the incidence of adverse events; secondary measures included number of Gd-enhanced lesions, confirmed relapses and EDSS changes. Except age, baseline demographic characteristics were well matched in both treatment arms. Both treatments were safe and well tolerated. M-GA induction produced an 89% greater reduction (relative risk (RR) = 0.11, 95% confidence interval (CI): 0.04 0.36, p = 0.0001) in the number of Gd-enhancing lesions at months 6 and 9 and a 70% reduction (RR = 0.30, 95% CI: 0.11-0.86, p = 0.0147) at months 12 and 15 versus GA alone. Mean relapse rates were 0.16 and 0.32 in the M-GA and GA groups, respectively. Short-term immunosuppression with mitoxantrone followed by daily GA for up to 15 months was found to be safe and effective, with an early and sustained decrease in MRI disease activity. PMID- 18424480 TI - Increased IL-10 mRNA and IL-23 mRNA expression in multiple sclerosis: interferon beta treatment increases IL-10 mRNA expression while reducing IL-23 mRNA expression. AB - BACKGROUND: Interferon (IFN)-beta therapy in multiple sclerosis (MS) has been suggested to promote a deviation from T lymphocyte production of pathogenic Th1 cytokines to less detrimental Th2 cytokines, but this is still controversial. We studied patterns of in vivo blood mononuclear cell (MNC) and whole blood cytokine and transcription factor mRNA expression before and during IFN-beta therapy in MS. METHODS: Twenty patients with relapsing-remitting MS were sampled before and after 3 months of treatment with IFN-beta along with 15 healthy volunteers. An additional 39 patients and 50 healthy volunteers served to confirm initial findings. mRNA was analyzed by real-time reverse transcriptase polymerase chain reaction (PCR). RESULTS: We found elevated expression of interleukin (IL)-23 and IL-10 in untreated MS patients. IFN-beta therapy increased IL-10 and decreased IL 23 expression independently of any Th1 or Th2 cytokines. The largest changes in cytokine mRNA levels occurred early (~9-12 h) after an IFN-beta injection. CONCLUSION: We found no evidence of a Th1- or Th2-mRNA-promoting effect of IFN beta therapy. The therapeutic effect of IFN-beta is more likely attributable to the induction of the regulatory cytokine IL-10. The elevated IL-23 mRNA levels in MS patients are noteworthy in view of the newly discovered IL-23-driven Th17 T cell subset, which is crucial in animal models of MS. Since IFN-beta therapy resulted in decreased IL-23 mRNA levels, the Th17 axis could be another target of IFN-beta therapy. PMID- 18424481 TI - Apneusis responding to buspirone in multiple sclerosis. AB - Apneusis is a disturbance of respiratory rhythm characterized by severely prolonged inspiratory effort, and is caused by bilateral lesions in the dorsal pons. In humans it is most commonly caused by pontine infarction and has rarely been reported in multiple sclerosis (MS). Here we report on a patient with MS who developed episodic apneusis which responded to treatment with buspirone, a serotonin type 1A receptor agonist. PMID- 18424482 TI - Axonal loss in the retinal nerve fiber layer in patients with multiple sclerosis. AB - OBJECTIVE: To quantify axonal loss in the retinal nerve fiber layer (RNFL) in patients with multiple sclerosis (MS), with and without a history of optic neuritis, by means of ocular imaging technologies. METHODS: This cross-sectional study enrolled 50 patients with MS and 25 age- and sex-matched healthy controls. All patients underwent neurologic assessment and a complete ophthalmic examination that included visual acuity, visual field examination, optical coherence tomography (OCT), scanning laser polarimetry (GDx) and visual evoked potentials (VEPs). Visual parameters and RNFL measurements were evaluated in MS eyes with a prior optic neuritis episode (MS-ON), with no prior episode (MS-NON) and control subjects. RESULTS: There were significant differences (p < 0.05, analysis of variance) between MS-ON (n = 25 eyes) and healthy eyes (n = 25 eyes) for all RNFL parameters measured by OCT and GDx. Significant differences between MS-NON (n = 75 eyes) and healthy eyes were also found for most of these parameters. RNFL thickness in the temporal quadrant was the parameter with the greatest differences between groups (71.79 microm in healthy eyes, 60.29 microm in MS-NON and 53.92 microm in MS-ON, p < 0.0005). Although there was a highly significant but moderate correlation between RNFL thickness and duration of the disease, no correlation was observed between RNFL thickness and neurologic impairment (Expanded Disability Status Scale). CONCLUSIONS: Axonal loss was detected not only in MS eyes with a previous acute optic neuritis, but also in MS eyes with no known optic neuritis episode. Structural abnormalities correlate with functional assessments of the optic nerve. PMID- 18424483 TI - Which lap is the slowest? An analysis of 32 world mile record performances. AB - OBJECTIVES: The pacing strategies adopted by world-record breakers during the 1 mile footrace in order to evaluate different models for the biological basis of pacing was determined in this study. METHODS: Lap times in 32 world record performances were analysed. Average times for each of the four laps and as percentages of total race time were calculated. RESULTS: The slowest laps in 90% of races were either the second (34%) or the third (56%) laps. In only two (6%) records was the final lap the slowest, whereas in 24 (76%), it was either the fastest (38%) or the second fastest (38%) lap. Mean times for the second and third laps were both significantly slower than were times for the first or final laps, but there was no significant difference in times for the first and final lap. CONCLUSION: The finding that world record beaters run the final lap in their quickest mile races faster than the second and third laps matches findings for races at longer distances. The presence of this "end spurt" suggests that the pacing strategy is regulated "in anticipation" and is not purely the result of a developing "peripheral fatigue". PMID- 18424484 TI - Testing for maximum oxygen consumption has produced a brainless model of human exercise performance. PMID- 18424485 TI - Is your prescription of distance running shoes evidence-based? AB - OBJECTIVES: To determine whether the current practice of prescribing distance running shoes featuring elevated cushioned heels and pronation control systems tailored to the individual's foot type is evidence-based. DATA SOURCES: MEDLINE (1950-May 2007), CINAHL (1982-May 2007), EMBASE (1980-May 2007), PsychInfo (1806 May 2007), Cochrane Database of Systematic Reviews (2(nd) Quarter 2007), Cochrane Central Register of Controlled trials (2(nd) Quarter 2007), SPORTSDiscus (1985 May 2007) and AMED (1985-May 2007). REVIEW METHODS: English language articles were identified via keyword and medical subject headings (MeSH) searches of the above electronic databases. With these searches and the subsequent review process, controlled trials or systematic reviews were sought in which the study population included adult recreational or competitive distance runners, the exposure was distance running, the intervention evaluated was a running shoe with an elevated cushioned heel and pronation control systems individualised to the wearer's foot type, and the outcome measures included either running injury rates, distance running performance, osteoarthritis risk, physical activity levels, or overall health and wellbeing. The quality of these studies and their findings were then evaluated. RESULTS: No original research that met the study criteria was identified either directly or via the findings of the six systematic reviews identified. CONCLUSION: The prescription of this shoe type to distance runners is not evidence-based. PMID- 18424486 TI - Injuries during the 2006 Women's Rugby World Cup. AB - BACKGROUND: Although there have been reports of injury rates in men's World Cup rugby, there has been no research into injury rates in women's international rugby. OBJECTIVE: To determine the rate of injury at the 2006 Women's Rugby World Cup (WRWC). METHODS: All participating teams in the 2006 WRWC were enrolled prospectively in this study. Healthcare workers for each team collected data on training and game exposure and information on injuries. RESULTS: 55 players (16%) sustained at least one injury (10.0/1000 player hours); 4 players (1%) sustained two injuries. 45 injuries occurred during games (37.5/1000 player hours) and 14 injuries occurred during practice (12.5/1000 player hours), a statistically significant difference (RR = 12.5, 95% CI 6.9 to 22.8). The injury rate was 39.3/1000 player hours for the forwards and 42.2/1000 player hours for the backs (RR = 1.3, 95% CI 0.7 to 2.3). The front row had the highest injury rate (62.5/1000 player hours). 63.6% of injuries occurred during the tackle. Most injuries occurred to the neck, knee and head/face. The majority of injuries were sprains, muscle injuries and contusions. There were five fractures during the event and four reported concussions. CONCLUSION: Female players sustained a considerable rate of injury during the 2006 WRWC. Backs had a slightly higher rate of injury than forwards, but the players most often injured were in the front row. The neck/cervical spine and knee were the most commonly injured regions, followed by the head and face. This is the first study to examine injury rates in female rugby players at a World Cup and reports valuable data in this population. PMID- 18424487 TI - Patellofemoral pain syndrome: a review on the associated neuromuscular deficits and current treatment options. AB - BACKGROUND: Patellofemoral pain syndrome (PFPS) is a common clinical presentation. Various neuromuscular factors have been reported to contribute to its aetiology. STUDY DESIGN: Systematic review METHODS: A literature search was carried out from 1998 up to December 2007. Eligible studies were those that: (1) examined the effects of hip strengthening in subjects with PFPS; (2) examined the effects of physiotherapy treatment aimed at restoring muscle balance between the vastus medialis oblique (VMO) and vastus lateralis (VL) in subjects with PFPS; (3) examined the effect of taping on electromyogram (EMG) muscle amplitude in subjects with PFPS; and (4) compared the effects of open versus closed kinetic chain exercises in the treatment of subjects with PFPS. RESULTS AND CONCLUSION: No randomised controlled trials exist to support the use of hip joint strengthening in subjects with PFPS. Physiotherapy treatment programmes appear to be an efficacious method of improving quadriceps muscle imbalances. Further studies are required to determine the true efficacy of therapeutic patellar taping. Both open and closed kinetic chain exercises are appropriate forms of treatment for subjects with PFPS. PMID- 18424488 TI - Tumor suppressor p53 slides on DNA with low friction and high stability. AB - The p53 protein, a transcription factor of key importance in tumorigenesis, is suggested to diffuse one-dimensionally along DNA via its C-terminal domain, a process that is proposed to regulate gene activation both positively and negatively. There has been no direct observation of p53 moving along DNA, however, and little is known about the mechanism and rate of its translocation. Here, we use single-molecule techniques to visualize, in real time, the one dimensional diffusion of p53 along DNA. The one-dimensional diffusion coefficient is measured to be close to the theoretical limit, indicative of movement along a free energy landscape with low activation barriers. We further investigate the mechanism of translocation and determine that p53 is capable of sliding--moving along DNA while in continuous contact with the duplex, rather than through a series of hops between nearby bases. PMID- 18424489 TI - Fast fluorescence laser tracking microrheometry, II: quantitative studies of cytoskeletal mechanotransduction. AB - Fluorescence laser tracking microrheometry (FLTM) is what we believe to be a novel method able to assess the local, frequency-dependent mechanical properties of living cells with nanometer spatial sensitivity at speeds up to 50 kHz. In an earlier article, we described the design, development, and optimization phases of the FLTM before reporting its performances in a variety of viscoelastic materials. In the work presented here, we demonstrate the suitability of FLTM to study local cellular rheology and obtain values for the storage and loss moduli G'(omega) and G''(omega) of fibroblasts consistent with past literature. We further establish that chemically induced cytoskeletal disruption is accompanied by reduced cellular stiffness and viscosity. Next, we provide a systematic study of some experimental variables that may critically influence microrheology measurements. First, we interrogate and justify the relevance of bead endocytosis as a method of cellular internalization of 1-microm probes in FLTM. Second, we show that as sample temperature increases, FLTM findings are elevated toward higher frequencies. Third, we confirm that relevant bead sizes (1 and 2 microm) have no effect on FLTM measurements. Fourth, we report the lack of influence of bead coatings (antiintegrin, antitransferrin, antidystroglycan, or uncoated tracers were surveyed) on their rheological readouts. Finally, we demonstrate the potential of FLTM in studying how substratum rigidity regulates cellular rheological properties. Interestingly, multiple, coupled strain relaxation mechanisms can be observed separated by two plateau moduli. Although these observations can be partly explained by rheological theories describing entangled actin filaments, there is a clear need to extend existing microrheology models to the cytoskeleton, including potentially important factors such as network geometry and remodeling. PMID- 18424490 TI - Endothelin receptor dimers evaluated by FRET, ligand binding, and calcium mobilization. AB - Endothelin-1 (ET-1) mediates physiological responses via endothelin A (ET(A)) and B (ET(B)) receptors, which may form homo- and heterodimers with unknown function. Here, we investigated ET-receptor dimerization using fluorescence resonance energy transfer (FRET) between receptors tagged with CFP (donor) and receptors tagged with tetracysteine-FlAsH (fluorescein arsenical hairpin) (acceptor) expressed in HEK293 cells. FRET efficiencies were 15%, 22%, and 27% for ET(A)/ET(A), ET(B)/ET(B), and ET(A)/ET(B), respectively, and dimerization was further supported by coimmunoprecipitation. For all dimer pairs, the natural but nonselective ligand ET-1 rapidly (50%, but did not detectably reduce coimmunoprecipitation. ET-1 stimulated a transient increase in intracellular Ca(2+) ([Ca(2+)](i)) lasting 1-2 min for both homodimer pairs, and these ET-1 actions on FRET and [Ca(2+)](i) elevation were blocked by the appropriate subtype-selective antagonist. In contrast, ET(A)/ET(B) heterodimers mediated a sustained [Ca(2+)](i) increase lasting >10 min, and required a combination of ET(A) and ET(B) antagonists to block the observed FRET and [Ca(2+)](i) responses. The sensitive CFP/FlAsH FRET assay used here provides new insights into endothelin-receptor dimer function, and represents a unique approach to characterize G-protein-coupled receptor oligomers, including their pharmacology. PMID- 18424491 TI - Mitochondrial swelling measurement in situ by optimized spatial filtering: astrocyte-neuron differences. AB - Mitochondrial swelling is a hallmark of mitochondrial dysfunction, and is an indicator of the opening of the mitochondrial permeability transition pore. We introduce here a novel quantitative in situ single-cell assay of mitochondrial swelling based on standard wide-field or confocal fluorescence microscopy. This morphometric technique quantifies the relative diameter of mitochondria labeled by targeted fluorescent proteins. Fluorescence micrographs are spatial bandpass filtered transmitting either high or low spatial frequencies. Mitochondrial swelling is measured by the fluorescence intensity ratio of the high- to low frequency filtered copy of the same image. We have termed this fraction the "thinness ratio". The filters are designed by numeric optimization for sensitivity. We characterized the thinness ratio technique by modeling microscopic image formation and by experimentation in cultured cortical neurons and astrocytes. The frequency domain image processing endows robustness and subresolution sensitivity to the thinness ratio technique, overcoming the limitations of shape measurement approaches. The thinness ratio proved to be highly sensitive to mitochondrial swelling, but insensitive to fission or fusion of mitochondria. We found that in situ astrocytic mitochondria swell upon short term uncoupling or inhibition of oxidative phosphorylation, whereas such responses are absent in cultured cortical neurons. PMID- 18424492 TI - Multifrequency forcing of a Hopf oscillator model of the inner ear. AB - In response to a sound stimulus, the inner ear emits sounds called otoacoustic emissions. While the exact mechanism for the production of otoacoustic emissions is not known, active motion of individual hair cells is thought to play a role. Two possible sources for otoacoustic emissions, both localized within individual hair cells, include somatic motility and hair bundle motility. Because physiological models of each of these systems are thought to be poised near a Hopf bifurcation, the dynamics of each can be described by the normal form for a system near a Hopf bifurcation. Here we demonstrate that experimental results from three-frequency suppression experiments can be predicted based on the response of an array of noninteracting Hopf oscillators tuned at different frequencies. This supports the idea that active motion of individual hair cells contributes to active processing of sounds in the ear. Interestingly, the model suggests an explanation for differing results recorded in mammals and nonmammals. PMID- 18424493 TI - Identification of the first steps in charge separation in bacterial photosynthetic reaction centers of Rhodobacter sphaeroides by ultrafast mid infrared spectroscopy: electron transfer and protein dynamics. AB - Time-resolved visible pump/mid-infrared (mid-IR) probe spectroscopy in the region between 1600 and 1800 cm(-1) was used to investigate electron transfer, radical pair relaxation, and protein relaxation at room temperature in the Rhodobacter sphaeroides reaction center (RC). Wild-type RCs both with and without the quinone electron acceptor Q(A), were excited at 600 nm (nonselective excitation), 800 nm (direct excitation of the monomeric bacteriochlorophyll (BChl) cofactors), and 860 nm (direct excitation of the dimer of primary donor (P) BChls (P(L)/P(M))). The region between 1600 and 1800 cm(-1) encompasses absorption changes associated with carbonyl (C=O) stretch vibrational modes of the cofactors and protein. After photoexcitation of the RC the primary electron donor P excited singlet state (P*) decayed on a timescale of 3.7 ps to the state P(+)B(L)(-) (where B(L) is the accessory BChl electron acceptor). This is the first report of the mid-IR absorption spectrum of P(+)B(L)(-); the difference spectrum indicates that the 9 keto C=O stretch of B(L) is located around 1670-1680 cm(-1). After subsequent electron transfer to the bacteriopheophytin H(L) in approximately 1 ps, the state P(+)H(L)(-) was formed. A sequential analysis and simultaneous target analysis of the data showed a relaxation of the P(+)H(L)(-) radical pair on the approximately 20 ps timescale, accompanied by a change in the relative ratio of the P(L)(+) and P(M)(+) bands and by a minor change in the band amplitude at 1640 cm(-1) that may be tentatively ascribed to the response of an amide C=O to the radical pair formation. We conclude that the drop in free energy associated with the relaxation of P(+)H(L)(-) is due to an increased localization of the electron hole on the P(L) half of the dimer and a further consequence is a reduction in the electrical field causing the Stark shift of one or more amide C=O oscillators. PMID- 18424494 TI - Tunable blinking kinetics of cy5 for precise DNA quantification and single nucleotide difference detection. AB - Fluorescence correlation spectroscopy (FCS) can resolve the intrinsic fast blinking kinetics (FBKs) of fluorescent molecules that occur on the order of microseconds. These FBKs can be heavily influenced by the microenvironments in which the fluorescent molecules are contained. In this work, FCS is used to monitor the dynamics of fluorescence emission from Cy5 labeled on DNA probes. We found that the FBKs of Cy5 can be tuned by having more or less unpaired guanines (upG) and thymines (upT) around the Cy5 dye. The observed FBKs of Cy5 are found to predominantly originate from the isomerization and back-isomerization processes of Cy5, and Cy5-nucleobase interactions are shown to slow down these processes. These findings lead to a more precise quantification of DNA hybridization using FCS analysis, in which the FBKs play a major role rather than the diffusion kinetics. We further show that the alterations of the FBKs of Cy5 on probe hybridization can be used to differentiate DNA targets with single nucleotide differences. This discrimination relies on the design of a probe target-probe DNA three-way-junction, whose basepairing configuration can be altered as a consequence of a single-nucleotide substitution on the target. Reconfiguration of the three-way-junction alters the Cy5-upG or Cy5-upT interactions, therefore resulting in a measurable change in Cy5 FBKs. Detection of single-nucleotide variations within a sequence selected from the Kras gene is carried out to validate the concept of this new method. PMID- 18424495 TI - Single molecule fluorescence analysis of branch migration of holliday junctions: effect of DNA sequence. AB - The Holliday junction is a central intermediate in various genetic processes including homologous, site-specific recombination and DNA replication. Recent single molecule FRET experiments led to the model for branch migration as a stepwise stochastic process in which the branch migration hop is terminated by the folding of the junction. In this article, we studied the effect of the sequence on Holliday junction dynamics and branch migration process. We show that a GC pair placed at the border of the homologous region almost prevents the migration into this position. At the same time, insertion of a GC pair into the middle of the AT tract does not show this effect, however when the junction folds at this position, it resides at this position much longer time in comparison to the folding at AT pairs. Two contiguous GC pairs do not block migration as well and generally manifest the same effect as one GC pair--the junction when it folds resides at these positions for a relatively long time. The same elevated residence time was obtained for the design with the homology region that consists of only GC pairs. These data suggest a model for branch migration in which the sequence modulates the overall stochastic process of the junction dynamics and branch migration by the variability of the time that the junction dwells before making a migration hop. PMID- 18424496 TI - Geometry of the nucleosomal DNA superhelix. AB - Nucleosome stability is largely an indirect measure of DNA sequence based on the material properties of DNA and the ability of a sequence to assume the required left-handed superhelical conformation. Here we focus attention only on the geometry of the superhelix and present two distinct mathematical expressions that rely on the DNA helical parameters (Shift, Slide, Rise, Tilt, Roll, Twist). One representation requires torsion for superhelix formation; the other requires shear. To compare these mathematical expressions to experimental data we develop a strategy for Fourier-filtering the helical parameters that identifies necessary and sufficient conditions to achieve a high-resolution model of the nucleosome superhelix. We apply this filtering strategy to 24 high-resolution structures of the nucleosome and demonstrate that all structures have a highly conserved distribution of Roll, Slide and Twist that involves two length scales. One length scale spans the entire length of nucleosomal DNA. The other is associated with the helix repeat. Our strategy also enables us to identify ground state or simple nucleosomes and altered nucleosome structures. These results form a basis for characterizing structural variations in the emerging family of nucleosome structures and a method for further developing structure-based models of nucleosome stability. PMID- 18424497 TI - Regulation of membrane proteins by dietary lipids: effects of cholesterol and docosahexaenoic acid acyl chain-containing phospholipids on rhodopsin stability and function. AB - Purified bovine rhodopsin was reconstituted into vesicles consisting of 1 stearoyl-2-oleoyl phosphatidylcholine or 1-stearoyl-2-docosahexaenoyl phosphatidylcholine with and without 30 mol % cholesterol. Rhodopsin stability was examined using differential scanning calorimetry (DSC). The thermal unfolding transition temperature (T(m)) of rhodopsin was scan rate-dependent, demonstrating the presence of a rate-limited component of denaturation. The activation energy of this kinetically controlled process (E(a)) was determined from DSC thermograms by four separate methods. Both T(m) and E(a) varied with bilayer composition. Cholesterol increased the T(m) both the presence and absence of docosahexaenoic acid acyl chains (DHA). In contrast, cholesterol lowered E(a) in the absence of DHA, but raised E(a) in the presence of 20 mol % DHA-containing phospholipid. The relative acyl chain packing order was determined from measurements of diphenylhexatriene fluorescence anisotropy decay. The T(m) for thermal unfolding was inversely related to acyl chain packing order. Rhodopsin kinetic stability (E(a)) was reduced in highly ordered or disordered membranes. Maximal kinetic stability was found within the range of acyl chain order found in native bovine rod outer segment disk membranes. The results demonstrate that membrane composition has distinct effects on the thermal versus kinetic stabilities of membrane proteins, and suggests that a balance between membrane constituents with opposite effects on acyl chain packing, such as DHA and cholesterol, may be required for maximum protein stability. PMID- 18424498 TI - Unloaded speed of shortening in voltage-clamped intact skeletal muscle fibers from wt, mdx, and transgenic minidystrophin mice using a novel high-speed acquisition system. AB - Skeletal muscle unloaded shortening has been indirectly determined in the past. Here, we present a novel high-speed optical tracking technique that allows recording of unloaded shortening in single intact, voltage-clamped mammalian skeletal muscle fibers with 2-ms time resolution. L-type Ca(2+) currents were simultaneously recorded. The time course of shortening was biexponential: a fast initial phase, tau(1), and a slower successive phase, tau(2,) with activation energies of 59 kJ/mol and 47 kJ/mol. Maximum unloaded shortening speed, v(u,max), was faster than that derived using other techniques, e.g., approximately 14.0 L(0) s(-1) at 30 degrees C. Our technique also allowed direct determination of shortening acceleration. We applied our technique to single fibers from C57 wild type, dystrophic mdx, and minidystrophin-expressing mice to test whether unloaded shortening was affected in the pathophysiological mechanism of Duchenne muscular dystrophy. v(u,max) and a(u,max) values were not significantly different in the three strains, whereas tau(1) and tau(2) were increased in mdx fibers. The results were complemented by myosin heavy and light chain (MLC) determinations that showed the same myosin heavy chain IIA profiles in the interossei muscles from the different strains. In mdx muscle, MLC-1f was significantly increased and MLC-2f and MLC-3f somewhat reduced. Fast initial active shortening seems almost unaffected in mdx muscle. PMID- 18424499 TI - DNA overstretching in the presence of glyoxal: structural evidence of force induced DNA melting. AB - When a long DNA molecule is stretched beyond its B-form contour length, a transition occurs in which its length increases by a factor of 1.7, with very little force increase. A quantitative model was proposed to describe this transition as force-induced melting, where double-stranded DNA is converted into single-stranded DNA. The force-induced melting model accurately describes the thermodynamics of DNA overstretching as a function of solution conditions and in the presence of DNA binding ligands. An alternative explanation suggests a transformation into S-DNA, a double-stranded form which preserves the interstrand base pairing. To determine the extent to which DNA base pairs are exposed to solution during the transition, we held DNA overstretched to different lengths within the transition in the presence of glyoxal. If overstretching involved strand separation, then force-melted basepairs would be glyoxal-modified, thus essentially permanently single-stranded. Subsequent stretches confirm that a significant fraction of the DNA melted by force is permanently melted. This result demonstrates that DNA overstretching is accompanied by a disruption of the DNA helical structure, including a loss of hydrogen bonding. PMID- 18424500 TI - Salt dependence of nucleic acid hairpin stability. AB - Single-stranded junctions/loops are frequently occurring structural motifs in nucleic acid structures. Due to the polyanionic nature of the nucleic acid backbone, metal ions play a crucial role in the loop stability. Here we use the tightly bound ion theory, which can account for the possible ion correlation and ensemble (fluctuation) effects, to predict the ion-dependence of loop and stem loop (hairpin) free energies. The predicted loop free energy is a function of the loop length, the loop end-to-end distance, and the ion (Na(+) and Mg(2+) in this study) concentrations. Based on the statistical mechanical calculations, we derive a set of empirical formulas for the loop thermodynamic parameters as functions of Na(+) and Mg(2+) concentrations. For three specific types of loops, namely, hairpin, bulge, and internal loops, the predicted free energies agree with the experimental data. Further applications of these empirical formulas to RNA and DNA hairpin stability lead to good agreements with the available experimental data. Our results indicate that the ion-dependent loop stability makes significant contribution to the overall ion-dependence of the hairpin stability. PMID- 18424501 TI - Docking of liposomes to planar surfaces mediated by trans-SNARE complexes. AB - Soluble N-ethylmaleimide-sensitive factor attachment protein receptors (SNAREs) play a key role in membrane fusion in the secretory pathway. In vitro, SNAREs spontaneously assemble into helical SNARE complexes with the transmembrane domains at the C-terminal end. During fusion, SNAREs are thought to bridge the two membranes and assemble in a zipper-like fashion, pulling the membranes together and initiating fusion. However, it is not clear to what extent SNARE assembly contributes to membrane attachment and membrane fusion. Using the neuronal SNAREs synaptobrevin (VAMP), SNAP-25, and syntaxin as examples, we show here that liposomes containing synaptobrevin firmly attach to planar surfaces containing immobilized syntaxin. Attachment requires the formation of SNARE complexes because it is dependent on the presence of SNAP-25. Binding is competed for by soluble SNARE fragments, with noncognate SNAREs such as endobrevin (VAMP8), VAMP4, and VAMP7 (Ti-VAMP) being effective but less potent in some cases. Furthermore, although SNAP-23 is unable to substitute for SNAP-25 in the attachment assay, it forms complexes of comparable stability and is capable of substituting in liposome fusion assays. Vesicle attachment is initiated by SNARE assembly at the N-terminal end of the helix bundle. We conclude that SNAREs can indeed form stable trans-complexes that result in vesicle attachment if progression to fusion is prevented, further supporting the zipper model of SNARE function. PMID- 18424502 TI - Rate of mixing controls rate and outcome of autocatalytic processes: theory and microfluidic experiments with chemical reactions and blood coagulation. AB - This article demonstrates that the rate of mixing can regulate the rate and outcome of both biological and nonbiological autocatalytic reaction systems that display a threshold response to the concentration of an activator. Plug-based microfluidics was used to control the timing of reactions, the rate of mixing, and surface chemistry in blood clotting and its chemical model. Initiation of clotting of human blood plasma required addition of a critical concentration of thrombin. Clotting could be prevented by rapid mixing when thrombin was added near the critical concentration, and mixing also affected the rate of clotting when thrombin was added at concentrations far above the critical concentration in two clinical clotting assays for human plasma. This phenomenon was modeled by a simple mechanism--local and global competition between the clotting reaction, which autocatalytically produces an activator, and mixing, which removes the activator. Numerical simulations showed that the Damkohler number, which describes this competition, predicts the effects of mixing. Many biological systems are controlled by thresholds, and these results shed light on the dynamics of these systems in the presence of spatial heterogeneities and provide simple guidelines for designing and interpreting experiments with such systems. PMID- 18424504 TI - Line tensions, correlation lengths, and critical exponents in lipid membranes near critical points. AB - Membranes containing a wide variety of ternary mixtures of high chain-melting temperature lipids, low chain-melting temperature lipids, and cholesterol undergo lateral phase separation into coexisting liquid phases at a miscibility transition. When membranes are prepared from a ternary lipid mixture at a critical composition, they pass through a miscibility critical point at the transition temperature. Since the critical temperature is typically on the order of room temperature, membranes provide an unusual opportunity in which to perform a quantitative study of biophysical systems that exhibit critical phenomena in the two-dimensional Ising universality class. As a critical point is approached from either high or low temperature, the scale of fluctuations in lipid composition, set by the correlation length, diverges. In addition, as a critical point is approached from low temperature, the line tension between coexisting phases decreases to zero. Here we quantitatively evaluate the temperature dependence of line tension between liquid domains and of fluctuation correlation lengths in lipid membranes to extract a critical exponent, nu. We obtain nu = 1.2 +/- 0.2, consistent with the Ising model prediction nu = 1. We also evaluate the probability distributions of pixel intensities in fluorescence images of membranes. From the temperature dependence of these distributions above the critical temperature, we extract an independent critical exponent of beta = 0.124 +/- 0.03, which is consistent with the Ising prediction of beta = 1/8. PMID- 18424503 TI - A fluorescence-based technique to construct size distributions from single-object measurements: application to the extrusion of lipid vesicles. AB - We report a novel approach to quantitatively determine complete size distributions of surface-bound objects using fluorescence microscopy. We measure the integrated intensity of single particles and relate it to their size by taking into account the object geometry and the illumination profile of the microscope, here a confocal laser scanning microscope. Polydisperse (as well as monodisperse) size distributions containing objects both below and above the optical resolution of the microscope are recorded and analyzed. The data is collected online within minutes, which allows the user to correlate the size of an object with the response from any given fluorescence-based biochemical assay. We measured the mean diameter of extruded fluorescently labeled lipid vesicles using the proposed method, dynamic light scattering, and cryogenic transmission electron microscopy. The three techniques were in excellent agreement, measuring the same values within 7-9%. Furthermore we demonstrated here, for the first time that we know of, the ability to determine the full size distribution of polydisperse samples of nonextruded lipid vesicles. Knowledge of the vesicle size distribution before and after extrusion allowed us to propose an empirical model to account for the effect of extrusion on the complete size distribution of vesicle samples. PMID- 18424505 TI - Cavity-creating mutations in Pseudomonas aeruginosa azurin: effects on protein dynamics and stability. AB - Changes in flexibility and structural stability of Pseudomonas aeruginosa azurin in response to cavity-creating mutations were probed by the phosphorescence emission of Trp-48, which was deeply buried in the compact hydrophobic core of the macromolecule, and by measurements of guanidinum hydrochloride unfolding, respectively. Replacement of the bulky side chains Phe-110, Phe-29, and Tyr-108 with the smaller Ala introduced cavities at different distances from the hydrophobic core. The phosphorescence lifetime (tau(0)) of Trp-48, buried inside the protein core, and the acrylamide quenching rate constant (k(q)) were used to monitor local and global flexibility changes induced by the introduction of the cavity. The results of this work demonstrate the following: 1), the effect on core flexibility of the insertion of cavities is not correlated readily to the distance of the cavity from the core; 2), the protein global flexibility results are related to the cavity distance from the packed core of the macromolecule; and 3), the increase in protein flexibility does not correspond necessarily to a comparable destabilizing effect of some mutations. PMID- 18424506 TI - Identification of a 2244 base pair interstitial deletion within the human ESR1 gene in the Spanish population. AB - BACKGROUND: The ESR1 AGATA haplotype is composed of five markers located within introns 5 and 6 of the human oestrogen receptor 1 (ESR1) gene. This haplotype has been studied in several male urogenital tract anomalies and male infertility. In one of these studies, a deviation from Hardy-Weinberg equilibrium (DHW) was found for the ESR1 AGATA marker rs3020375 in two groups of healthy controls. In the present study, we investigated whether the observed DHW is caused by structural variants present within the ESR1 gene. PARTICIPANTS: 229 family units achieving pregnancy through assisted reproductive technologies (n = 129) or by natural means (n = 100), 2465 general Spanish population controls and 162 men with idiopathic infertility. MAIN OUTCOME MEASUREMENTS: Segregation analyses of genetic markers in family units and case-control genetic association studies. RESULTS: We identified a new interstitial deletion of 2244 base pairs within intron 6 of the human ESR1 gene as the cause for the observed DHW. This new variant presents a 10% allelic frequency in the general Spanish population and it is associated with idiopathic male infertility (OR = 1.51; p = 0.03). The percentage of infertile couples in which both members carried the ESR1 deletion (10.08%) was also a higher than expected value of 6% (p = 0.03). CONCLUSIONS: We have characterised a novel structural variation in human ESR1 gene. The available data indicate a deleterious action of the ESR1 deletion in both male and couple fertility. The potential effects of this deletion on other oestrogen-related diseases need to be determined. PMID- 18424507 TI - Genome-wide linkage scan for loci of musical aptitude in Finnish families: evidence for a major locus at 4q22. AB - BACKGROUND: Music perception and performance are comprehensive human cognitive functions and thus provide an excellent model system for studying human behaviour and brain function. However, the molecules involved in mediating music perception and performance are so far uncharacterised. OBJECTIVE: To unravel the biological background of music perception, using molecular and statistical genetic approaches. METHODS: 15 Finnish multigenerational families (with a total of 234 family members) were recruited via a nationwide search. The phenotype of all family members was determined using three tests used in defining musical aptitude: a test for auditory structuring ability (Karma Music test; KMT) commonly used in Finland, and the Seashore pitch and time discrimination subtests (SP and ST respectively) used internationally. We calculated heritabilities and performed a genome-wide variance components-based linkage scan using genotype data for 1113 microsatellite markers. RESULTS: The heritability estimates were 42% for KMT, 57% for SP, 21% for ST and 48% for the combined music test scores. Significant evidence of linkage was obtained on chromosome 4q22 (LOD 3.33) and suggestive evidence of linkage at 8q13-21 (LOD 2.29) with the combined music test scores, using variance component linkage analyses. The major contribution of the 4q22 locus was obtained for the KMT (LOD 2.91). Interestingly, a positive LOD score of 1.69 was shown at 18q, a region previously linked to dyslexia (DYX6) using combined music test scores. CONCLUSION: Our results show that there is a genetic contribution to musical aptitude that is likely to be regulated by several predisposing genes or variants. PMID- 18424508 TI - Screening BRCA1 and BRCA2 unclassified variants for splicing mutations using reverse transcription PCR on patient RNA and an ex vivo assay based on a splicing reporter minigene. AB - BACKGROUND: Many unclassified variants (UV) of BRCA1 or BRCA2 may have an effect on pre-mRNA splicing. Patient blood samples suitable for RNA extraction are not always available for testing UVs at the RNA level. METHODS: Analyses of RNA from patient peripheral blood were performed, using a one-step reverse transcriptase PCR (RT-PCR) protocol, and were compared with an ex vivo splicing assay based on PCR-amplified patient DNA inserted into a splicing reporter minigene. Using both methods 20 variants found in 17 patients were examined. RESULTS: Data from patient RNA and from the minigene assay were fully concordant, but the ex vivo splicing assay, which is monoallelic, clarified several ambiguities in the patient RNA data. Two intronic variants induced strong splicing defects: BRCA1 c.4987-5T-->A (IVS16-5T-->A) induced exon 17 skipping and BRCA2 c.316+5G-->C (IVS3+5G-->C) induced complete skipping of exon 3. Of the exonic variants, BRCA2 c.7805G-->C (p.Arg2602Thr), at the last base of exon 16, induced both exon skipping and activation of a cryptic exonic donor site, and BRCA2 c.8023A-->G (p.Ile2675Val) generated a strong donor site within exon 18. These four variants were thus classified as pathogenic, because of the total absence of a normal transcript from the corresponding allele. Variant BRCA2 c.9501+3A-->T (IVS25+3A- >T) induced incomplete skipping of exon 25, suggesting a mutation with incomplete penetrance, and BRCA2 c.8257_8259del (p.Leu2753del) modified the alternative splicing of exons 17 and 18. CONCLUSIONS: We show that functional analysis using a splicing reporter minigene is sensitive and specific, and should be used for initial screening of potential splicing defects, especially when patient RNA is not readily available. PMID- 18424509 TI - Justice, equality and solidarity: the limits of the right to health care in Israel. PMID- 18424510 TI - Candida albicans Tup1 is involved in farnesol-mediated inhibition of filamentous growth induction. AB - Candida albicans is a dimorphic fungus that can interconvert between yeast and filamentous forms. Its ability to regulate morphogenesis is strongly correlated with virulence. Tup1, a transcriptional repressor, and the signaling molecule farnesol are both capable of negatively regulating the yeast to filamentous conversion. Based on this overlap in function, we tested the hypothesis that the cellular response to farnesol involves, in part, the activation of Tup1. Tup1 functions with the DNA binding proteins Nrg1 and Rfg1 as a transcription regulator to repress the expression of hypha-specific genes. The tup1/tup1 and nrg1/nrg1 mutants, but not the rfg1/rfg1 mutant, failed to respond to farnesol. Treatment of C. albicans cells with farnesol caused a small but consistent increase in both TUP1 mRNA and protein levels. Importantly, this increase corresponds with the commitment point, beyond which added farnesol no longer blocks germ tube formation, and it correlates with a strong decrease in the expression of two Tup1-regulated hypha-specific genes, HWP1 and RBT1. Tup1 probably plays a direct role in the response to farnesol because farnesol suppresses the haploinsufficient phenotype of a TUP1/tup1 heterozygote. Farnesol did not affect EFG1 (a transcription regulator of filament development), NRG1, or RFG1 mRNA levels, demonstrating specific gene regulation in response to farnesol. Furthermore, the tup1/tup1 and nrg1/nrg1 mutants produced 17- and 19-fold more farnesol, respectively, than the parental strain. These levels of excess farnesol are sufficient to block filamentation in a wild-type strain. Our data are consistent with the role of Tup1 as a crucial component of the response to farnesol in C. albicans. PMID- 18424511 TI - A folded and functional protein domain in an amyloid-like fibril. AB - The effect of the polypeptide environment on polyalanine-induced fibril formation was investigated with amyloidogenic fragments from PAPBN1, a nuclear protein controlling polyadenylation. Mutation-caused extensions of the natural 10 alanine sequence up to maximally 17 alanines result in fibril formation of PABPN1 and the development of the disease oculopharyngeal muscular dystrophy (OPMD). We explored the influence of fibril formation on the structure and function of a one-domain protein linked to the fibril-forming part of PABPN1. The well-characterized, stably folded, one-domain protein, cold-shock protein CspB from Bacillus subtilis, was fused either to the C terminus of the entire N-terminal domain of PABPN1 or directly to peptides consisting of 10 or 17 alanine residues. The fusion protein between the N-terminal domain of PABPN1 and CspB formed fibrils in which the structure and activity of CspB were retained. In the fibrils formed by fusions in which the polyalanine sequence was directly linked to CspB, CspB was unfolded. These results indicate that the folded conformation and the function of a protein domain can be maintained in amyloid-like fibrils, and that the distance between this domain and the fibril plays an important role. PMID- 18424512 TI - Distinguishing the cross-beta spine arrangements in amyloid fibrils using FRET analysis. AB - The recently published microcrystal structures of amyloid fibrils from small peptides greatly enhanced our understanding of the atomic-level structure of the amyloid fibril. However, only a few amyloid fibrils can form microcrystals. The dansyl-tryptophan fluorescence resonance energy transfer (FRET) pair was shown to be able to detect the inter-peptide arrangement of the Transthyretin (105-115) amyloid fibril. In this study, we combined the known microcrystal structures with the corresponding FRET efficiencies to build a model for amyloid fibril structure classification. We found that fibrils with an antiparallel structural arrangement gave the largest FRET signal, those with a parallel arrangement gave the lowest FRET signal, and those with a mixed arrangement gave a moderate FRET signal. This confirms that the amyloid fibril structure patterns can be classified based on the FRET efficiency. PMID- 18424513 TI - Crystal structures of barley thioredoxin h isoforms HvTrxh1 and HvTrxh2 reveal features involved in protein recognition and possibly in discriminating the isoform specificity. AB - H-type thioredoxins (Trxs) constitute a particularly large Trx sub-group in higher plants. Here, the crystal structures are determined for the two barley Trx h isoforms, HvTrxh1 and HvTrxh2, in the partially radiation-reduced state to resolutions of 1.7 A, and for HvTrxh2 in the oxidized state to 2.0 A. The two Trxs have a sequence identity of 51% and highly similar fold and active-site architecture. Interestingly, the four independent molecules in the crystals of HvTrxh1 form two relatively large and essentially identical protein-protein interfaces. In each interface, a loop segment of one HvTrxh1 molecule is positioned along a shallow hydrophobic groove at the primary nucleophile Cys40 of another HvTrxh1 molecule. The association mode can serve as a model for the target protein recognition by Trx, as it brings the Met82 Cgamma atom (gamma position as a disulfide sulfur) of the bound loop segment in the proximity of the Cys40 thiol. The interaction involves three characteristic backbone-backbone hydrogen bonds in an antiparallel beta-sheet-like arrangement, similar to the arrangement observed in the structure of an engineered, covalently bound complex between Trx and a substrate protein, as reported by Maeda et al. in an earlier paper. The occurrence of an intermolecular salt bridge between Glu80 of the bound loop segment and Arg101 near the hydrophobic groove suggests that charge complementarity plays a role in the specificity of Trx. In HvTrxh2, isoleucine corresponds to this arginine, which emphasizes the potential for specificity differences between the coexisting barley Trx isoforms. PMID- 18424514 TI - Characterization of ubiquitin and ubiquitin-like-protein isopeptidase activities. AB - Conjugation or deconjugation of ubiquitin (Ub) or ubiquitin-like proteins (UBLs) to or from cellular proteins is a multifaceted and universal means of regulating cellular physiology, controlling the lifetime, localization, and activity of many critical proteins. Deconjugation of Ub or UBL from proteins is performed by a class of proteases called isopeptidases. Herein is described a readily quantifiable novel isopeptidase assay platform consisting of Ub or UBL fused to the reporter enzyme phospholipase A(2) (PLA(2)). Isopeptidase activity releases PLA(2), which cleaves its substrate, generating a signal that is linear with deubiquitylase (DUB) concentration and is able to discriminate DUB, deSUMOylase, deNEDDylase, and deISGylase activities. The power and sensitivity of the UBL PLA(2) assay are demonstrated by its ability to differentiate the contrasting deISGylase and DUB activities of two coronavirus proteases: severe acute respiratory syndrome papain-like protease (SARS-CoV PLpro) and NL63 CoV papain like protease 2 (PLP2). Furthermore, direct comparisons with the current Ub-7 amino-4-methylcoumarin (Ub-AMC) assay demonstrated that the Ub-PLA(2) assay is an effective tool for characterizing modulators of isopeptidase activity. This observation was expanded by profiling the inhibitory activity of the nonselective isopeptidase inhibitor NSC 632839 against DUBs and deSUMOylases. Taken together, these studies illustrate the utility of the reporter-based approach to measuring isopeptidase activity. PMID- 18424515 TI - Glucosamine found as a substituent of both phosphate groups in Bordetella lipid A backbones: role of a BvgAS-activated ArnT ortholog. AB - Endotoxins are amphipathic lipopolysaccharides (LPSs), major constituents of the outer membrane of gram-negative bacteria. They consist of a lipid region, covalently linked to a core oligosaccharide, to which may be linked a repetitive glycosidic chain carrying antigenic determinants. Most of the biological activities of endotoxins have been associated with the lipid moiety of the molecule: unique to gram-negative bacteria, LPS is a ligand of the mammalian TLR4 MD2-CD14 pathogen recognition receptor complex. Lipid A preparations are often heterogeneous with respect to both the numbers and the lengths of fatty acids and the natures of substituents on the phosphate groups when present. The variants can significantly affect host immune responses. Nine species in the Bordetella genus have been described, and the fine LPS structures of seven of them have been published. In this report, lipids A from Bordetella pertussis Tohama I and B. bronchiseptica strain 4650 were further characterized and revealed to have a glucosamine substituting both lipid A phosphate groups of the diglucosamine backbone. These substitutions have not been previously described for bordetellae. Moreover, a B. pertussis transposon mutation that maps within a gene encoding a Bordetella ArnT (formerly PmrK) glycosyl transferase ortholog does not carry this substitution, thus providing a genetic basis for the modification. Reverse transcriptase PCR of this locus showed that it is Bvg regulated, suggesting that the ability of Bordetella to modify lipid A via this glucosamine modification is a potential virulence trait. PMID- 18424517 TI - ApoFnr binds as a monomer to promoters regulating the expression of enterotoxin genes of Bacillus cereus. AB - Bacillus cereus Fnr is a member of the Crp/Fnr (cyclic AMP-binding protein/fumarate nitrate reduction regulatory protein) family of helix-turn-helix transcriptional regulators. It is essential for the expression of hbl and nhe enterotoxin genes independently of the oxygen tension in the environment. We studied aerobic Fnr binding to target sites in promoters regulating the expression of enterotoxin genes. B. cereus Fnr was overexpressed and purified as either a C-terminal His-tagged (Fnr(His)) fusion protein or an N-terminal fusion protein tagged with the Strep-tag (IBA BioTAGnology) ((Strep)Fnr). Both recombinant Fnr proteins were produced as apoforms (clusterless) and occurred as mixtures of monomers and oligomers in solution. However, apoFnr(His) was mainly monomeric, while apo(Strep)Fnr was mainly oligomeric, suggesting that the His tagged C-terminal extremity may interfere with oligomerization. The oligomeric state of apo(Strep)Fnr was dithiothreitol sensitive, underlining the importance of a disulfide bridge for apoFnr oligomerization. Electrophoretic mobility shift assays showed that monomeric apoFnr, but not oligomeric apoFnr, bound to specific sequences located in the promoter regions of the enterotoxin regulators fnr, resDE, and plcR and the structural genes hbl and nhe. The question of whether apoFnr binding is regulated in vivo by redox-dependent oligomerization is discussed. PMID- 18424516 TI - Bacteriophage P22 antitermination boxB sequence requirements are complex and overlap with those of lambda. AB - Transcription antitermination in phages lambda and P22 uses N proteins that bind to similar boxB RNA hairpins in regulated transcripts. In contrast to the lambda N-boxB interaction, the P22 N-boxB interaction has not been extensively studied. A nuclear magnetic resonance structure of the P22 N peptide boxB(left) complex and limited mutagenesis have been reported but do not reveal a consensus sequence for boxB. We have used a plasmid-based antitermination system to screen boxBs with random loops and to test boxB mutants. We find that P22 N requires boxB to have a GNRA-like loop with no simple requirements on the remaining sequences in the loop or stem. U:A or A:U base pairs are strongly preferred adjacent to the loop and appear to modulate N binding in cooperation with the loop and distal stem. A few GNRA-like hexaloops have moderate activity. Some boxB mutants bind P22 and lambda N, indicating that the requirements imposed on boxB by P22 N overlap those imposed by lambda N. Point mutations can dramatically alter boxB specificity between P22 and lambda N. A boxB specific for P22 N can be mutated to lambda N specificity by a series of single mutations via a bifunctional intermediate, as predicted by neutral theories of evolution. PMID- 18424518 TI - YscP and YscU switch the substrate specificity of the Yersinia type III secretion system by regulating export of the inner rod protein YscI. AB - Pathogenic yersiniae utilize a type III secretion system to inject antihost factors, called Yops, directly into the cytosol of eukaryotic cells. The Yops are injected via a needle-like structure, comprising the YscF protein, on the bacterial surface. While the needle is being assembled, Yops cannot be secreted. YscP and YscU switch the substrate specificity of the secretion system to enable Yop export once the needle attains its proper length. Here, we demonstrate that the inner rod protein YscI plays a critical role in substrate specificity switching. We show that YscI is secreted by the type III secretion system and that YscI secretion by a yscP mutant is abnormally elevated. Furthermore, we show that mutations in the cytoplasmic domain of YscU reduce YscI secretion by the yscP null strain. We also demonstrate that mutants expressing one of three forms of YscI (those with mutations Q84A, L87A, and L96A) secrete substantial amounts of Yops yet exhibit severe defects in needle formation. In the absence of YscP, mutants with the same changes in YscI assemble needles but are unable to secrete Yops. Together, these results suggest that the formation of the inner rod, not the needle, is critical for substrate specificity switching and that YscP and YscU exert their effects on substrate export by controlling the secretion of YscI. PMID- 18424519 TI - Formate acts as a diffusible signal to induce Salmonella invasion. AB - To infect an animal host, Salmonella enterica serovar Typhimurium must penetrate the intestinal epithelial barrier. This process of invasion requires a type III secretion system encoded within Salmonella pathogenicity island I (SPI1). We found that a mutant with deletions of the acetate kinase and phosphotransacetylase genes (ackA-pta) was deficient in invasion and SPI1 expression but that invasion gene expression was completely restored by supplying medium conditioned by growth of the wild-type strain, suggesting that a signal produced by the wild type, but not by the ackA-pta mutant, was required for invasion. This mutant also excreted 68-fold-less formate into the culture medium, and the addition of sodium formate to cultures restored both the expression of SPI1 and the invasion of cultured epithelial cells by the mutant. The effect of formate was pH dependent, requiring a pH below neutrality, and studies in mice showed that the distal ileum, the preferred site of Salmonella invasion in this species, had the appropriate formate concentration and pH to elicit invasion, while the cecum contained no detectable formate. Furthermore, we found that formate affected the major regulators of SPI1, hilA and hilD, but that the primary routes of formate metabolism played no role in its activity as a signal. PMID- 18424520 TI - Functional analysis of the protein machinery required for transport of lipopolysaccharide to the outer membrane of Escherichia coli. AB - Lipopolysaccharide (LPS) is an essential component of the outer membrane (OM) in most gram-negative bacteria, and its structure and biosynthetic pathway are well known. Nevertheless, the mechanisms of transport and assembly of this molecule at the cell surface are poorly understood. The inner membrane (IM) transport protein MsbA is responsible for flipping LPS across the IM. Additional components of the LPS transport machinery downstream of MsbA have been identified, including the OM protein complex LptD/LptE (formerly Imp/RlpB), the periplasmic LptA protein, the IM-associated cytoplasmic ATP binding cassette protein LptB, and LptC (formerly YrbK), an essential IM component of the LPS transport machinery characterized in this work. Here we show that depletion of any of the proteins mentioned above leads to common phenotypes, including (i) the presence of abnormal membrane structures in the periplasm, (ii) accumulation of de novo-synthesized LPS in two membrane fractions with lower density than the OM, and (iii) accumulation of a modified LPS, which is ligated to repeating units of colanic acid in the outer leaflet of the IM. Our results suggest that LptA, LptB, LptC, LptD, and LptE operate in the LPS assembly pathway and, together with other as-yet-unidentified components, could be part of a complex devoted to the transport of LPS from the periplasmic surface of the IM to the OM. Moreover, the location of at least one of these five proteins in every cellular compartment suggests a model for how the LPS assembly pathway is organized and ordered in space. PMID- 18424521 TI - Identification of a novel trimeric autotransporter adhesin in the cryptic genospecies of Haemophilus. AB - Haemophilus biotype IV strains belonging to the recently recognized Haemophilus cryptic genospecies are an important cause of maternal genital tract and neonatal systemic infections and initiate infection by colonizing the genital or respiratory epithelium. To gain insight into the mechanism of Haemophilus cryptic genospecies colonization, we began by examining prototype strain 1595 and three other strains for adherence to genital and respiratory epithelial cell lines. Strain 1595 and two of the three other strains demonstrated efficient adherence to all of the cell lines tested. With a stably adherent variant of strain 1595, we generated a Mariner transposon library and identified 16 nonadherent mutants. All of these mutants lacked surface fibers and contained an insertion in the same open reading frame, which encodes a 157-kDa protein designated Cha for cryptic haemophilus adhesin. Analysis of the predicted amino acid sequence of Cha revealed the presence of an N-terminal signal peptide and a C-terminal domain bearing homology to YadA-like and Hia-like trimeric autotransporters. Examination of the C-terminal 120 amino acids of Cha demonstrated mobility as a trimer on sodium dodecyl sulfate-polyacrylamide gel electrophoresis and the capacity to present the passenger domain of the Hia trimeric autotransporter on the bacterial surface. Southern analysis revealed that the gene that encodes Cha is conserved among clinical isolates of the Haemophilus cryptic genospecies and is absent from the closely related species Haemophilus influenzae. We speculate that Cha is important in the pathogenesis of disease due to the Haemophilus cryptic genospecies and is in part responsible for the apparent tissue tropism of this organism. PMID- 18424522 TI - Transcriptional interference and repression modulate the conjugative ability of the symbiotic plasmid of Rhizobium etli. AB - Bacteria of the order Rhizobiales are able to establish nitrogen-fixing symbioses with legumes. Commonly, genes for symbiosis are harbored on large symbiotic plasmids. Although the transfer of symbiotic plasmids is commonly detected in nature, there are few experimentally characterized examples. In Rhizobium etli, the product of rctA inhibits the conjugation of the symbiotic plasmid by reducing the transcription of the virB operon. rctA is transcribed divergently from this operon, and its product is predicted to have a DNA binding domain. In the present study, using DNase I footprinting and binding assays, we demonstrated the specific binding of RctA to the virB operon promoter. A 9-bp motif in the spacer region of this promoter (the rctA binding motif box) and the presence of a functional -10 region were critical elements for RctA binding. Transcriptional fusion analyses revealed that the elimination of either element provoked a relief of RctA-mediated repression. These data support a model in which RctA inhibits the access of the RNA polymerase to the virB promoter. Interestingly, rctA expression levels were modulated by transcriptional interference from transcripts emanating from the virB promoter. This phenomenon adds another level of regulation for this system, thus revealing a novel mechanism of plasmid transfer regulation in the Rhizobiales. PMID- 18424523 TI - Mutations in the tacF gene of clinical strains and laboratory transformants of Streptococcus pneumoniae: impact on choline auxotrophy and growth rate. AB - The nutritional requirement that Streptococcus pneumoniae has for the aminoalcohol choline as a component of teichoic and lipoteichoic acids appears to be exclusive to this prokaryote. A mutation in the tacF gene, which putatively encodes an integral membrane protein (possibly, a teichoic acid repeat unit transporter), has been recently identified as responsible for generating a choline-independent phenotype of S. pneumoniae (M. Damjanovic, A. S. Kharat, A. Eberhardt, A. Tomasz, and W. Vollmer, J. Bacteriol. 189:7105-7111, 2007). We now report that Streptococcus mitis can grow in choline-free medium, as previously illustrated for Streptococcus oralis. While we confirmed the finding by Damjanovic et al. of the involvement of TacF in the choline dependence of the pneumococcus, the genetic transformation of S. pneumoniae R6 by using S. mitis SK598 DNA and several PCR-amplified tacF fragments suggested that a minimum of two mutations were required to confer improved fitness to choline-independent S. pneumoniae mutants. This conclusion is supported by sequencing results also reported here that indicate that a spontaneous mutant of S. pneumoniae (strain JY2190) able to proliferate in the absence of choline (or analogs) is also a double mutant for the tacF gene. Microscopic observations and competition experiments during the cocultivation of choline-independent strains confirmed that a minimum of two amino acid changes were required to confer improved fitness to choline-independent pneumococcal strains when growing in medium lacking any aminoalcohol. Our results suggest complex relationships among the different regions of the TacF teichoic acid repeat unit transporter. PMID- 18424524 TI - Conserved residues Asp16 and Pro24 of TnaC-tRNAPro participate in tryptophan induction of Tna operon expression. AB - In Escherichia coli, interactions between the nascent TnaC-tRNA(Pro) peptidyl tRNA and the translating ribosome create a tryptophan binding site in the ribosome where bound tryptophan inhibits TnaC-tRNA(Pro) cleavage. This inhibition delays ribosome release, thereby inhibiting Rho factor binding and action, resulting in increased tna operon transcription. Replacing Trp12 of TnaC with any other amino acid residue was previously shown to prevent tryptophan binding and induction of tna operon expression. Genome-wide comparisons of TnaC amino acid sequences identify Asp16 and Pro24, as well as Trp12, as highly conserved TnaC residues. Replacing these residues with other residues was previously shown to influence tryptophan induction of tna operon expression. In this study, in vitro analyses were performed to examine the potential roles of Asp16 and Pro24 in tna operon induction. Replacing Asp16 or Pro24 of TnaC of E. coli with other amino acids established that these residues are essential for free tryptophan binding and inhibition of TnaC-tRNA(Pro) cleavage at the peptidyl transferase center. Asp16 and Pro24 are in fact located in spatial positions corresponding to critical residues of AAP, another ribosome regulatory peptide. Sparsomycin methylation protection studies further suggested that segments of 23S RNA were arranged differently in ribosomes bearing TnaCs with either the Asp16Ala or the Pro24Ala change. Thus, features of the amino acid sequence of TnaC of the nascent TnaC-tRNA(Pro) peptidyl-tRNA, in addition to the presence of Trp12, are necessary for the nascent peptide to create a tryptophan binding/inhibition site in the translating ribosome. PMID- 18424525 TI - UpaG, a new member of the trimeric autotransporter family of adhesins in uropathogenic Escherichia coli. AB - The ability of Escherichia coli to colonize both intestinal and extraintestinal sites is driven by the presence of specific virulence factors, among which are the autotransporter (AT) proteins. Members of the trimeric AT adhesin family are important virulence factors for several gram-negative pathogens and mediate adherence to eukaryotic cells and extracellular matrix (ECM) proteins. In this study, we characterized a new trimeric AT adhesin (UpaG) from uropathogenic E. coli (UPEC). Molecular analysis of UpaG revealed that it is translocated to the cell surface and adopts a multimeric conformation. We demonstrated that UpaG is able to promote cell aggregation and biofilm formation on abiotic surfaces in CFT073 and various UPEC strains. In addition, UpaG expression resulted in the adhesion of CFT073 to human bladder epithelial cells, with specific affinity to fibronectin and laminin. Prevalence analysis revealed that upaG is strongly associated with E. coli strains from the B2 and D phylogenetic groups, while deletion of upaG had no significant effect on the ability of CFT073 to colonize the mouse urinary tract. Thus, UpaG is a novel trimeric AT adhesin from E. coli that mediates aggregation, biofilm formation, and adhesion to various ECM proteins. PMID- 18424526 TI - The critical role of embC in Mycobacterium tuberculosis. AB - Arabinan polymers are major components of the cell wall in Mycobacterium tuberculosis and are involved in maintaining its structure, as well as playing a role in host-pathogen interactions. In particular, lipoarabinomannan (LAM) has multiple immunomodulatory effects. In the nonpathogenic species Mycobacterium smegmatis, EmbC has been identified as a key arabinosyltransferase involved in the incorporation of arabinose into LAM, and an embC mutant is viable but lacks LAM. In contrast, we demonstrate here that in M. tuberculosis, embC is an essential gene under normal growth conditions, suggesting a more crucial role for LAM in the pathogenic mycobacteria. M. tuberculosis EmbC has an activity similar to that of M. smegmatis EmbC, since we were able to complement an embC mutant of M. smegmatis with embC(Mtb), confirming that it encodes a functional arabinosyltransferase. In addition, we observed that the size of LAM produced in M. smegmatis was dependent on the level of expression of embC(Mtb). Northern analysis revealed that embC is expressed as part of a polycistronic message encompassing embC and three upstream genes. The promoter region for this transcript was identified and found to be up-regulated in stationary phase but down-regulated during hypoxia-induced nonreplicating persistence. In conclusion, we have identified one of the key genes involved in LAM biosynthesis in M. tuberculosis and confirmed its essential role in this species. PMID- 18424527 TI - The ArgP protein stimulates the Klebsiella pneumoniae gdhA promoter in a lysine sensitive manner. AB - The lysine-sensitive factor that binds to the upstream region of the Klebsiella pneumoniae gdhA promoter and stimulates gdhA transcription during growth in minimal medium has been proposed to be the K. pneumoniae ArgP protein (M. R. Nandineni, R. S. Laishram, and J. Gowrishankar, J. Bacteriol. 186:6391-6399, 2004). A knockout mutation of the K. pneumoniae argP gene was generated and used to assess the roles of exogenous lysine and argP in the regulation of the gdhA promoter. Disruption of argP reduced the strength and the lysine-dependent regulation of the gdhA promoter. Electrophoretic mobility shift assays using crude extracts prepared from wild-type and argP-defective strains indicted the presence of an argP-dependent factor whose ability to bind the gdhA promoter was lysine sensitive. DNase I footprinting studies using purified K. pneumoniae ArgP protein indicated that ArgP bound the region that lies approximately 50 to 100 base pairs upstream of the gdhA transcription start site in a manner that was sensitive to the presence of lysine. Substitutions within the region bound by ArgP affected the binding of ArgP to the gdhA promoter region in vitro and the argP-dependent stimulation of the gdhA promoter in vivo. These observations suggest that elevated intracellular levels of lysine reduce the affinity of ArgP for its binding site at the gdhA promoter, preventing ArgP from binding to and stimulating transcription from the promoter in vivo. PMID- 18424528 TI - How family, community, and work structured high blood pressure accounts: from African Americans in Washington State. AB - High blood pressure is one of the most often researched, yet least understood health disparities among African Americans. This descriptive, critical discourse analysis examined how family and community demographics and paid and unpaid work structured participants' accounts of high blood pressure experiences in Washington State. Thirty-seven urban-dwelling African American women (n = 17) and men (n = 20) in Washington State enrolled in the study from 2000-2001. Reports about stress, concerns, worry, loneliness, and paid and unpaid work were given in semi-structured interviews. Analysis results are embedded within three major themes: (a) Aware, But Not Informed, (b) Negotiating Self, Kin and Community Health, and (c) Distant Lives, Distant Love. Knowledge of life factors influencing African Americans' high blood pressure appraisals will help develop context-specific health programs focused on their concerns. PMID- 18424529 TI - Hypericum and nurses: a comprehensive literature review on the efficacy of St. John's Wort in the treatment of depression. AB - PURPOSE: Many patients look to complementary and alternative medicine for a herbal solution to depression. This literature review summarizes recently published research on the treatment of depression using St. John's wort (Hypericum perforatum). CONCLUSIONS: The compounds in St. John's wort herbal preparations are more effective than placebo and, in several studies, more effective than common antidepressant medications in treating minor depression. However, the efficacy of St. John's wort for treating major depression, cyclothymia, or bipolar disorder is less evident. Although some studies are promising in the treatment of these major disorders, research support is lacking, and it is a controversial aspect of Hypericum therapy. PRACTICE IMPLICATIONS: As with any herbal treatment, risks from adverse reactions and drug interactions exist. Providers have an ethical and legal obligation to stay current in knowledge and to provide useful, accurate information to patients. PMID- 18424530 TI - Competing isogenic Campylobacter strains exhibit variable population structures in vivo. AB - Consumption of poultry contaminated with Campylobacter jejuni is a risk factor for human gastrointestinal disease. The rational development of control strategies for Campylobacter within chickens requires an understanding of the colonization process at the molecular and population levels, both within and between hosts. Experiments employing competing strains of Campylobacter have been used to investigate colonization. Implicit in these studies is the assumption that the behavior of competing strains is reproducible between experiments. Variability in the recovery of mutants from the chicken gastrointestinal tract during signature-tagged mutagenesis studies demonstrated that this is not always the case. To further investigate this phenomenon in the absence of confounding factors due to phenotypic differences between mutants, we constructed individually identifiable wild-type isogenic tagged strains (WITS) that have indistinguishable phenotypes in pure culture. By using mixtures of WITS, it is possible to monitor the relative amounts of subpopulations of essentially wild type bacteria. Using a 2-week-old chicken model of colonization, we observed unpredictable variations in population structure both within and between experiments, even in the simplest case of two competing strains. This variation occurred both when birds were simultaneously infected with two WITS and when birds inoculated with different WITS were cohoused. We present evidence for founder effects during initial colonization with subsequent bird-to-bird transmission. We suggest that these and phenotypic variation contribute to the observed variability. These factors render simple models of colonization which do not take them into account inappropriate for Campylobacter and impact the planning and interpretation of competition experiments using this organism. PMID- 18424531 TI - Genetic structure and biology of Xylella fastidiosa strains causing disease in citrus and coffee in Brazil. AB - Xylella fastidiosa is a vector-borne, plant-pathogenic bacterium that causes disease in citrus (citrus variegated chlorosis [CVC]) and coffee (coffee leaf scorch [CLS]) plants in Brazil. CVC and CLS occur sympatrically and share leafhopper vectors; thus, determining whether X. fastidiosa isolates can be dispersed from one crop to another and cause disease is of epidemiological importance. We sought to clarify the genetic and biological relationships between CVC- and CLS-causing X. fastidiosa isolates. We used cross-inoculation bioassays and microsatellite and multilocus sequence typing (MLST) approaches to determine the host range and genetic structure of 26 CVC and 20 CLS isolates collected from different regions in Brazil. Our results show that citrus and coffee X. fastidiosa isolates are biologically distinct. Cross-inoculation tests showed that isolates causing CVC and CLS in the field were able to colonize citrus and coffee plants, respectively, but not the other host, indicating biological isolation between the strains. The microsatellite analysis separated most X. fastidiosa populations tested on the basis of the host plant from which they were isolated. However, recombination among isolates was detected and a lack of congruency among phylogenetic trees was observed for the loci used in the MLST scheme. Altogether, our study indicates that CVC and CLS are caused by two biologically distinct strains of X. fastidiosa that have diverged but are genetically homogenized by frequent recombination. PMID- 18424532 TI - Development and experimental validation of a predictive threshold cycle equation for quantification of virulence and marker genes by high-throughput nanoliter volume PCR on the OpenArray platform. AB - Development of quantitative PCR (QPCR) assays typically requires extensive screening within and across a given species to ensure specific detection and lucid identification among various pathogenic and nonpathogenic strains and to generate standard curves. To minimize screening requirements, multiple virulence and marker genes (VMGs) were targeted simultaneously to enhance reliability, and a predictive threshold cycle (C(T)) equation was developed to calculate the number of starting copies based on an experimental C(T). The empirical equation was developed with Sybr green detection in nanoliter-volume QPCR chambers (OpenArray) and tested with 220 previously unvalidated primer pairs targeting 200 VMGs from 30 pathogens. A high correlation (R(2) = 0.816) was observed between the predicted and experimental C(T)s based on the organism's genome size, guanine and cytosine (GC) content, amplicon length, and stability of the primer's 3' end. The performance of the predictive C(T) equation was tested using 36 validation samples consisting of pathogenic organisms spiked into genomic DNA extracted from three environmental waters. In addition, the primer success rate was dependent on the GC content of the target organisms and primer sequences. Targeting multiple assays per organism and using the predictive C(T) equation are expected to reduce the extent of the validation necessary when developing QPCR arrays for a large number of pathogens or other targets. PMID- 18424533 TI - Associations between antimicrobial resistance genes in fecal generic Escherichia coli isolates from cow-calf herds in western Canada. AB - The objective of this study was to examine associations among the genetic determinants of antimicrobial resistance (AMR) in 207 fecal generic Escherichia coli isolates obtained from 77 cow-calf herds in western Canada. Twenty-three resistance genes corresponding to six different antimicrobial families were assessed using DNA hybridization and PCR. The most common resistance genes in the study sample (207 isolates) were sul2 (48.3%), tet(B) (45.4%), and ant(3'')-Ia (aadA1) (19.3%). Several statistically significant associations between the examined resistance genes were detected. The strongest associations observed were those between genes for resistance to chloramphenicol (catI) and trimethoprim (dhfrI) (odds ratio [OR] = 214; P = 0.0001), sulfonamide (sul1) and chloramphenicol (catI) (OR = 96.9; P = 0.0001), streptomycin [ant(3'')-Ia (aadA1)] and trimethoprim (dhfrI) (OR = 96.2; P = 0.0001), sulfonamide (sul1) and streptomycin [ant(3'')-Ia (aadA1)] (OR = 79.3; P = 0.0001), and tetracycline [tet(B)] and sulfonamides (sul2) (OR = 25.7; P = 0.0001). At least one of the resistance genes corresponding to each nonaminoglycoside family of antimicrobials examined in this study was associated with the two aminoglycoside resistance genes ant(3'')-Ia (aadA1) and aph(3')-Ia. The multiple, strong associations between genes and the diverse nature of the associations described in this study demonstrate the complexity of resistance gene selection in cow-calf herds and should be considered in the planning of AMR control practices for cow-calf operations. PMID- 18424535 TI - Biphenyl dioxygenase from an arctic isolate is not cold adapted. AB - Biphenyl dioxygenase from the psychrotolerant bacterium Pseudomonas sp. strain Cam-1 (BPDO(Cam-1)) was purified and found to have an apparent k(cat) for biphenyl of 1.1 +/- 0.1 s(-1) (mean +/- standard deviation) at 4 degrees C. In contrast, BPDO(LB400) from the mesophile Burkholderia xenovorans LB400 had no detectable activity at this temperature. At 57 degrees C, the half-life of the BPDO(Cam-1) oxygenase was less than half that of the BPDO(LB400) oxygenase. Nevertheless, BPDO(Cam-1) appears to be a typical Pseudomonas pseudoalcaligenes KF707-type dioxygenase. PMID- 18424534 TI - Characterization of a bacterial community in an abandoned semiarid lead-zinc mine tailing site. AB - Bacterial diversity in mine tailing microbial communities has not been thoroughly investigated despite the correlations that have been observed between the relative microbial diversity and the success of revegetation efforts at tailing sites. This study employed phylogenetic analyses of 16S rRNA genes to compare the bacterial communities present in highly disturbed, extremely (pH 2.7) and moderately (pH 5.7) acidic lead-zinc mine tailing samples from a semiarid environment with those from a vegetated off-site (OS) control sample (pH 8). Phylotype richness in these communities decreased from 42 in the OS control to 24 in the moderately acidic samples and 8 in the extremely acidic tailing samples. The clones in the extremely acidic tailing sample were most closely related to acidophiles, none of which were detected in the OS control sample. The comparison generated by this study between the bacteria present in extremely acidic tailing and that in moderately acidic tailing communities with those in an OS control soil provides a reference point from which to evaluate the successful restoration of mine tailing disposal sites by phytostabilization. PMID- 18424536 TI - Rapid screening of quorum-sensing signal N-acyl homoserine lactones by an in vitro cell-free assay. AB - A simple, sensitive, and rapid cell-free assay system was developed for detection of N-acyl homoserine lactone (AHL) autoinducers involved in bacterial quorum sensing (QS). The present approach improves upon previous whole-cell biosensor based approaches in its utilization of a cell-free assay approach to conduct bioassays. The cell-free assay was derived from the AHL biosensor bacterium Agrobacterium tumefaciens NTL4(pCF218)(pCF372), allowing the expression of beta galactosidase upon addition of exogenous AHLs. We have shown that beta galactosidase expression is possible in cell-free solution [lysate from Agrobacterium tumefaciens NTL4(pCF218)(pCF372) culture]. Assay detection limits with the use of chromogenic substrate X-Gal (5-bromo-4-chloro-3-indolyl-beta-D galactopyranoside) ranged from approximately 100 nM to 300 nM depending on the specific AHL. Replacement (of X-Gal) with the luminescent substrate Beta-Glo increased sensitivity to AHLs by 10-fold. A major advantage of the cell-free assay system is elimination of time-consuming steps for biosensor cell culture conditioning, which are required prior to whole-cell bioassays. This significantly reduced assay times from greater than 24 h to less than 3 h, while maintaining high sensitivity. Assay lysate may be prepared in bulk and stored ( 80 degrees C) over 6 months for future use. Finally, the present protocol may be adapted for use with other biosensor strains and be used in high-throughput AHL screening of bacteria or metagenomic libraries. PMID- 18424538 TI - Influence of environmental stress on distributions of times to first division in Escherichia coli populations, as determined by digital-image analysis of individual cells. AB - The distributions of times to first cell division were determined for populations of Escherichia coli stationary-phase cells inoculated onto agar media. This was accomplished by using automated analysis of digital images of individual cells growing on agar and calculation of the "box area ratio." Using approximately 300 cells per experiment, the mean time to first division and standard deviation for cells grown in liquid medium at 37 degrees C and inoculated on agar and incubated at 20 degrees C were determined as 3.0 h and 0.7 h, respectively. Distributions were observed to tail toward the higher values, but no definitive model distribution was identified. Both preinoculation stress by heating cultures at 50 degrees C and postinoculation stress by growth in the presence of higher concentrations of NaCl increased mean times to first division. Both stresses also resulted in an increase in the spread of the distributions that was proportional to the mean division time, the coefficient of variation being constant at approximately 0.2 in all cases. The "relative division time," which is the time to first division for individual cells expressed in terms of the cell size doubling time, was used as measure of the "work to be done" to prepare for cell division. Relative division times were greater for heat-stressed cells than for those growing under osmotic stress. PMID- 18424539 TI - Longitudinal study of the molecular epidemiology of Campylobacter jejuni in cattle on dairy farms. AB - Multilocus sequence typing (MLST), an accurate and phylogenetically robust characterization method for population studies of Campylobacter, was applied to Campylobacter jejuni isolates (n = 297) from the fecal samples of cattle from five dairy farms in Cheshire, United Kingdom, collected throughout 2003. The population dynamics of the C. jejuni strains, as identified by the occurrence of sequence types and clonal complexes, demonstrated variations within and between cattle populations over time. Three clonal lineages have emerged to predominate among the cattle isolates, namely, the ST-61 complex (24.2%), ST-21 complex (23.6%), and ST-42 complex (20.5%). This provided further evidence that the ST-61 clonal complex may present a cattle-adapted C. jejuni genotype. In addition, the ST-42 clonal complex may also represent an important cattle-associated genotype. Strong geographical associations for these genotypes were also found among the farms. This is the first longitudinal study and the largest study to date for C. jejuni involving cattle populations using MLST for accurate strain characterization. This study shows the important associations between cattle and C. jejuni clonal complexes ST-61, ST-21, and ST-42, and it suggests that cattle and/or dairy products are likely to be a source of the human Campylobacter gastroenteritis caused by such genotypes. The reported findings have significant implications for the design of effective intervention strategies for disease control and prevention. PMID- 18424537 TI - Population structure of the fish-pathogenic bacterium Flavobacterium psychrophilum. AB - Flavobacterium psychrophilum is currently one of the main bacterial pathogens hampering the productivity of salmonid farming worldwide, and its control mainly relies on antibiotic treatments. To better understand the population structure of this bacterium and its mode of evolution, we have examined the nucleotide polymorphisms at 11 protein-coding loci of the core genome in a set of 50 isolates. These isolates were selected to represent the broadest possible diversity, originating from 10 different host fish species and four continents. The nucleotide diversity between pairs of sequences amounted to fewer than four differences per kilobase on average, corresponding to a particularly low level of diversity, possibly indicative of a small effective-population size. The recombination rate, however, seemed remarkably high, and as a consequence, most of the isolates harbored unique combinations of alleles (33 distinct sequence types were resolved). The analysis also showed the existence of several clonal complexes with worldwide geographic distribution but marked association with particular fish species. Such an association could reflect preferential routes of transmission and/or adaptive niche specialization. The analysis provided no clues that the initial range of the bacterium was originally limited to North America. Instead, the historical record of the expansion of the pathogen may reflect the spread of a few clonal complexes. As a resource for future epidemiological surveys, a multilocus sequence typing website based on seven highly informative loci is available. PMID- 18424540 TI - Structure of the archaeal community of the rumen. PMID- 18424541 TI - Longitudinal study of antimicrobial resistance among Escherichia coli isolates from integrated multisite cohorts of humans and swine. AB - In a 3-year longitudinal study, we examined the relationship between the seasonal prevalence of antimicrobial-resistant (AR) Escherichia coli isolates from human wastewater and swine fecal samples and the following risk factors: the host species, the production type (swine), the vocation (human swine workers, non swine workers, and slaughter plant workers), and the season, in a multisite, vertically integrated swine and human population representative of a closed agri food system. Human and swine E. coli (n = 4,048 and 3,429, respectively) isolates from wastewater and fecal samples were tested for antimicrobial susceptibility, using the Sensititre broth microdilution system. There were significant (P < 0.05) differences among AR E. coli prevalence levels of (i) the host species, in which swine isolates were at higher risk for resistance to tetracycline, kanamycin, ceftiofur, gentamicin, streptomycin, chloramphenicol, sulfisoxazole, and ampicillin; (ii) the swine production group, in which purchased boars, nursery piglets, and breeding boars isolates had a higher risk of resistance to streptomycin and tetracycline; and iii) the vocation cohorts, in which swine worker cohort isolates exhibited lower sulfisoxazole and cefoxitin prevalence than the non-swine worker cohorts, while the slaughter plant worker cohort isolates exhibited elevated cefoxitin prevalence compared to that of non-swine workers. While a high variability was observed among seasonal samples over the 3 year period, no significant temporal trends were apparent. There were significant differences in the prevalence levels of multidrug-resistant isolates between host species, with swine at a higher risk of carrying multidrug-resistant strains than humans. Considering vocation, slaughter plant workers were at higher risk of exhibiting multidrug-resistant E. coli than non-swine workers. PMID- 18424542 TI - Chitin hydrolysis by Listeria spp., including L. monocytogenes. AB - Listeria spp., including the food-borne pathogen Listeria monocytogenes, are ubiquitous microorganisms in the environment and thus are difficult to exclude from food processing plants. The factors that contribute to their multiplication and survival in nature are not well understood, but the ability to catabolize various carbohydrates is likely to be very important. One major source of carbon and nitrogen in nature is chitin, an insoluble linear beta-1,4-linked polymer of N-acetylglucosamine (GlcNAc). Chitin is found in cell walls of fungi and certain algae, in the cuticles of arthropods, and in shells and radulae of molluscs. In the present study, we demonstrated that L. monocytogenes and other Listeria spp. are able to hydrolyze alpha-chitin. The chitinolytic activity is repressed by the presence of glucose in the medium, suggesting that chitinolytic activity is subjected to catabolite repression. Activity is also regulated by temperature and is higher at 30 degrees C than at 37 degrees C. In L. monocytogenes EGD, chitin hydrolysis depends on genes encoding two chitinases, lmo0105 (chiB) and lmo1883 (chiA), but not on a gene encoding a putative chitin binding protein (lmo2467). The chiB and chiA genes are phylogenetically related to various well characterized chitinases. The potential biological implications of chitinolytic activity of Listeria are discussed. PMID- 18424543 TI - Effect of exposure to UV-C irradiation and monochloramine on adenovirus serotype 2 early protein expression and DNA replication. AB - The mechanisms of adenovirus serotype 2 inactivation with either UV light (with a narrow emission spectrum centered at 254 nm) or monochloramine were investigated by assessing the potential inhibition of two key steps of the adenovirus life cycle, namely, E1A protein synthesis and viral genomic replication. E1A early protein synthesis was assayed by using immunoblotting, while the replication of viral DNA was analyzed by using slot blotting. Disinfection experiments were performed in phosphate buffer solutions at pH 8 and room temperature (UV) or 20 degrees C (monochloramine). Experimental results revealed that normalized E1A levels at 12 h postinfection (p.i.) were statistically the same as the corresponding decrease in survival ratio for both UV and monochloramine disinfection. Normalized DNA levels at 24 h p.i. were also found to be statistically the same as the corresponding decrease in survival ratio for monochloramine disinfection. In contrast, for UV disinfection, genomic DNA levels were much lower than E1A or survival ratios, possibly as a result of a delay in DNA replication for UV-treated virions compared to that for controls. Future efforts will determine the pre-E1A synthesis step in the adenovirus life cycle affected by exposure to UV and monochloramine, with the goal of identifying the viral molecular target of these two disinfectants. PMID- 18424544 TI - Characterization of the pattern of alphas1- and beta-casein breakdown and release of a bioactive peptide by a cell envelope proteinase from Lactobacillus delbrueckii subsp. lactis CRL 581. AB - The cell envelope-associated proteinases (CEPs) of the lactobacilli have key roles in bacterial nutrition and contribute to the development of the organoleptic properties of fermented milk products as well, as they can release bioactive health-beneficial peptides from milk proteins. The influence of the peptide supply, carbohydrate source, and osmolites on the CEP activity of the cheese starter Lactobacillus delbrueckii subsp. lactis CRL 581 was investigated. The CEP activity levels were controlled by the peptide content of the growth medium. The maximum activity was observed in a basal minimal defined medium, whereas in the presence of Casitone, Casamino Acids, or yeast extract, the synthesis of CEP was inhibited 99-, 70-, and 68-fold, respectively. The addition of specific di- or tripeptides containing branched-chain amino acids, such as leucylleucine, prolylleucine, leucylglycylglycine, or leucylproline, to the growth medium negatively affected CEP activity, whereas dipeptides without branched-chain amino acids had no effect on the enzyme's production. The carbon source and osmolites did not affect CEP activity. The CEP of L. delbrueckii subsp. lactis CRL 581 exhibited a mixed-type CEP(I/III) variant caseinolytic specificity. Mass-spectrometric screening of the main peptide peaks isolated by reverse-phase high-pressure liquid chromatography allowed the identification of 33 and 32 peptides in the alpha(s1)- and beta-casein hydrolysates, respectively. By characterizing the peptide sequence in these hydrolysates, a pattern of alpha(s1)- and beta-casein breakdown was defined and is reported herein, this being the first report for a CEP of L. delbrueckii subsp. lactis. In this pattern, a series of potentially bioactive peptides (antihypertensive and phosphopeptides) which are encrypted within the precursor protein could be visualized. PMID- 18424545 TI - Construction of a reporter vector system for in vivo analysis of promoter activity in Propionibacterium freudenreichii. AB - A beta-galactosidase reporter system for the analysis of promoter elements in Propionibacterium freudenreichii was designed. The pTD210 in vivo reporter vector was constructed using a promoterless lacZ gene from Bifidobacterium longum cloned into the pAMT1 plasmid. The utility of the pTD210 reporter vector was demonstrated by an investigation of six predicted promoters in P. freudenreichii. The system produced accurate and reproducible measurements that facilitated both promoter identification and the quantification of promoter activities. PMID- 18424546 TI - The cyclic AMP-dependent catabolite repression system of Serratia marcescens mediates biofilm formation through regulation of type 1 fimbriae. AB - The mechanisms by which environmental carbon sources regulate biofilm formation are poorly understood. This study investigates the roles of glucose and the catabolite repression system in Serratia marcescens biofilm formation. The abilities of this opportunistic pathogen to proliferate in a wide range of environments, to cause disease, and to resist antimicrobials are linked to its ability to form biofilms. We observed that growth of S. marcescens in glucose rich medium strongly stimulated biofilm formation, which contrasts with previous studies showing that biofilm formation is inhibited by glucose in Escherichia coli and other enteric bacteria. Glucose uptake is known to inversely mediate intracellular cyclic AMP (cAMP) synthesis through regulation of adenylate cyclase (cyaA) activity, which in turn controls fundamental processes such as motility, carbon utilization and storage, pathogenesis, and cell division in many bacteria. Here, we demonstrate that mutation of catabolite repression genes that regulate cAMP levels (crr and cyaA) or the ability to respond to cAMP (crp) confers a large increase in biofilm formation. Suppressor analysis revealed that phenotypes of a cAMP receptor protein (crp) mutant require the fimABCD operon, which is responsible for type 1 fimbria production. Consistently, fimA transcription and fimbria production were determined to be upregulated in a cyaA mutant background by using quantitative real-time reverse transcription-PCR and transmission electron microscopy analysis. The regulatory pathway by which environmental carbon sources influence cAMP concentrations to alter production of type 1 fimbrial adhesins establishes a novel mechanism by which bacteria control biofilm development. PMID- 18424547 TI - Minimal Escherichia coli cell for the most efficient production of ethanol from hexoses and pentoses. AB - To obtain an efficient ethanologenic Escherichia coli strain, we reduced the functional space of the central metabolic network, with eight gene knockout mutations, from over 15,000 pathway possibilities to 6 pathway options that support cell function. The remaining pathways, identified by elementary mode analysis, consist of four pathways with non-growth-associated conversion of pentoses and hexoses into ethanol at theoretical yields and two pathways with tight coupling of anaerobic cell growth with ethanol formation at high yields. Elimination of three additional genes resulted in a strain that selectively grows only on pentoses, even in the presence of glucose, with a high ethanol yield. We showed that the ethanol yields of strains with minimized metabolic functionality closely matched the theoretical predictions. Remarkably, catabolite repression was completely absent during anaerobic growth, resulting in the simultaneous utilization of pentoses and hexoses for ethanol production. PMID- 18424548 TI - Anaerobic and aerobic degradation of cyanophycin by the denitrifying bacterium Pseudomonas alcaligenes strain DIP1 and role of three other coisolates in a mixed bacterial consortium. AB - Four bacterial strains were isolated from a cyanophycin granule polypeptide (CGP) degrading anaerobic consortium, identified by 16S rRNA gene sequencing, and assigned to species of the genera Pseudomonas, Enterococcus, Clostridium, and Paenibacillus. The consortium member responsible for CGP degradation was assigned as Pseudomonas alcaligenes strain DIP1. The growth of and CGP degradation by strain DIP1 under anaerobic conditions were enhanced but not dependent on the presence of nitrate as an electron acceptor. CGP was hydrolyzed to its constituting beta-Asp-Arg dipeptides, which were then completely utilized within 25 and 4 days under anaerobic and aerobic conditions, respectively. The end products of CGP degradation by strain DIP1 were alanine, succinate, and ornithine as determined by high-performance liquid chromatography analysis. The facultative anaerobic Enterococcus casseliflavus strain ELS3 and the strictly anaerobic Clostridium sulfidogenes strain SGB2 were coisolates and utilized the beta-linked isodipeptides from the common pool available to the mixed consortium, while the fourth isolate, Paenibacillus odorifer strain PNF4, did not play a direct role in the biodegradation of CGP. Several syntrophic interactions affecting CGP degradation, such as substrate utilization, the reduction of electron acceptors, and aeration, were elucidated. This study demonstrates the first investigation of CGP degradation under both anaerobic and aerobic conditions by one bacterial strain, with regard to the physiological role of other bacteria in a mixed consortium. PMID- 18424549 TI - Blocking of bacterial biofilm formation by a fish protein coating. AB - Bacterial biofilm formation on inert surfaces is a significant health and economic problem in a wide range of environmental, industrial, and medical areas. Bacterial adhesion is generally a prerequisite for this colonization process and, thus, represents an attractive target for the development of biofilm-preventive measures. We have previously found that the preconditioning of several different inert materials with an aqueous fish muscle extract, composed primarily of fish muscle alpha-tropomyosin, significantly discourages bacterial attachment and adhesion to these surfaces. Here, this proteinaceous coating is characterized with regards to its biofilm-reducing properties by using a range of urinary tract infectious isolates with various pathogenic and adhesive properties. The antiadhesive coating significantly reduced or delayed biofilm formation by all these isolates under every condition examined. The biofilm-reducing activity did, however, vary depending on the substratum physicochemical characteristics and the environmental conditions studied. These data illustrate the importance of protein conditioning layers with respect to bacterial biofilm formation and suggest that antiadhesive proteins may offer an attractive measure for reducing or delaying biofilm-associated infections. PMID- 18424550 TI - Protection against hydrogen peroxide-mediated cytotoxicity in Friedreich's ataxia fibroblasts using novel iron chelators of the 2-pyridylcarboxaldehyde isonicotinoyl hydrazone class. AB - Iron-loading diseases remain an important problem because of the toxicity of iron catalyzed redox reactions. Iron loading occurs in the mitochondria of Friedreich's ataxia (FA) patients and may play a role in its pathogenesis. This suggests that iron chelation therapy could be useful. We developed previously the lipophilic iron chelators known as the 2-pyridylcarboxaldehyde isonicotinoyl hydrazone (PCIH) ligands and identified 2-pyridylcarboxaldehyde 2 thiophenecarboxyl hydrazone (PCTH) as the most promising analog. Hence, this study assessed the efficacy of PCTH and other PCIH analogs compared with various chelators, including deferiprone and desferrioxamine (DFO). Age- and sex-matched control and FA fibroblasts were preincubated with iron chelators and subsequently challenged with 50 microM H2O2 for up to 24 h. The current study demonstrates an interesting structure-activity relationship among the closely related PCIH series of ligands, with only PCTH being highly effective at preventing H2O2-induced cytotoxicity. PCTH increased FA fibroblast cell viability by up to 70%, whereas DFO rescued viability by 1 to 5% only. Hence, PCTH, which was well tolerated by cells was far more effective than DFO at preventing oxidative stress. It is noteworthy that kinetic studies demonstrated PCTH to rapidly penetrate cells to induce 59Fe efflux, whereas DFO, PCIH, 2-pyridylcarboxaldehyde benzoyl hydrazone, and 2-pyridylcarboxaldehyde m-bromobenzoyl hydrazone were far slower, indicating it is the rate of chelator permeation that is crucial for protection against H2O2. In addition, PCTH was found to be as effective as or more effective than conventional radical scavengers or the antioxidant idebenone (which has undergone clinical trials) at protecting cells against H2O2-mediated cytotoxicity. These findings further indicate the potential of PCTH for treatment of iron overload. PMID- 18424551 TI - N-palmitoyl glycine, a novel endogenous lipid that acts as a modulator of calcium influx and nitric oxide production in sensory neurons. AB - N-arachidonoyl glycine is an endogenous arachidonoyl amide that activates the orphan G protein-coupled receptor (GPCR) GPR18 in a pertussis toxin (PTX) sensitive manner and produces antinociceptive and antiinflammatory effects. It is produced by direct conjugation of arachidonic acid to glycine and by oxidative metabolism of the endocannabinoid anandamide. Based on the presence of enzymes that conjugate fatty acids with glycine and the high abundance of palmitic acid in the brain, we hypothesized the endogenous formation of the saturated N-acyl amide N-palmitoyl glycine (PalGly). PalGly was partially purified from rat lipid extracts and identified using nano-high-performance liquid chromatography/hybrid quadrupole time-of-flight mass spectrometry. Here, we show that PalGly is produced after cellular stimulation and that it occurs in high levels in rat skin and spinal cord. PalGly was up-regulated in fatty acid amide hydrolase knockout mice, suggesting a pathway for enzymatic regulation. PalGly potently inhibited heat-evoked firing of nociceptive neurons in rat dorsal horn. In addition, PalGly induced transient calcium influx in native adult dorsal root ganglion (DRG) cells and a DRG-like cell line (F-11). The effect of PalGly on the latter cells was characterized by strict structural requirements, PTX sensitivity, and dependence on the presence of extracellular calcium. PalGly-induced calcium influx was blocked by the nonselective calcium channel blockers ruthenium red, 1-(beta-[3-(4 methoxyphenyl)propoxy]-4-methoxyphenethyl)-1H-imidazole (SK&F96365), and La3+. Furthermore, PalGly contributed to the production of NO through calcium-sensitive nitric-oxide synthase enzymes present in F-11 cells and was inhibited by the nitric-oxide synthase inhibitor 7-nitroindazole. PMID- 18424552 TI - (2R)-[(4-Biphenylylsulfonyl)amino]-N-hydroxy-3-phenylpropionamide (BiPS), a matrix metalloprotease inhibitor, is a novel and potent activator of hypoxia inducible factors. AB - Hypoxia-inducible factors (HIFs) are unstable heterodimeric transcription factors and decisive elements for the transcriptional regulation of genes important in the adaptation to low-oxygen conditions. Hypoxia is the ubiquitous inducer of HIFs, stabilizing the alpha-subunit and permitting the formation of a functional HIF complex. Here, we identify (2R)-[(4-biphenylylsulfonyl)amino]-N-hydroxy-3 phenylpropionamide (BiPS), a commercially available metalloprotease-2 and -9 inhibitor, as a rapid and potent inducer of HIFs. We show that in different cell lines, BiPS induces the HIF-alpha subunit by inhibiting its degradation through stabilization of its labile oxygen-dependent degradation domain. This is achieved through the inhibition of HIF-1alpha hydroxylation. The HIF-1 complex, formed after BiPS treatment, is capable of DNA binding and activation of HIF target genes, including the expression of vascular endothelial growth factor. Because novel HIF activators have generated considerable interest in the possible treatment of different ischemic diseases, we believe that BiPS and derivative molecules could have strong therapeutic potential. PMID- 18424553 TI - Individually monitoring ligand-induced changes in the structure of the GABAA receptor at benzodiazepine binding site and non-binding-site interfaces. AB - The mechanisms by which the GABA and benzodiazepine (BZD) binding sites of the GABA-A receptor are allosterically coupled remain elusive. In this study, we separately monitored ligand-induced structural changes in the BZD binding site (alpha/gamma interface) and at aligned positions in the alpha/beta interface. alpha(1)His101 and surrounding residues were individually mutated to cysteine and expressed with wild-type beta2 and gamma2 subunits in Xenopus laevis oocytes. The accessibilities of introduced cysteines to modification by methanethiosulfonate ethylammonium (MTSEA)-Biotin were measured in the presence and absence of GABA site agonists, antagonists, BZDs, and pentobarbital. The presence of flurazepam or the BZD-site antagonist flumazenil (Ro15-1788) decreased the rate of modification of alpha(1)H101C at the BZD binding site. GABA and muscimol each increased MTSEA-Biotin modification of alpha(1)H101C located at the BZD-site, gabazine (SR-95531, a GABA binding site antagonist) decreased the rate, whereas pentobarbital had no effect. Modification of alpha(1)H101C at the alpha/beta interface was significantly slower than modification of alpha(1)H101C at the BZD site, and the presence of GABA or flurazepam had no effect on its accessibility, indicating the physicochemical environments of the alpha/gamma and alpha/beta interfaces are different. The data are consistent with the idea that GABA-binding site occupation by agonists causes a GABA binding cavity closure that is directly coupled to BZD binding cavity opening, and GABA-site antagonist binding causes a movement linked to BZD binding cavity closure. Pentobarbital binding/gating resulted in no observable movements in the BZD binding site near alpha(1)H101C, indicating that structural mechanisms underlying allosteric coupling between the GABA and BZD binding sites are distinct. PMID- 18424554 TI - Reciprocal regulation of dopamine D1 and D3 receptor function and trafficking by heterodimerization. AB - Colocalization of dopamine D1 (D1R) and D3 receptors (D3R) in specific neuronal populations suggests that their functional cross-talk might involve direct interactions. Here we report that the D1R coimmunoprecipitates with the D3R from striatal protein preparations, suggesting that they are clustered together in this region. Using bioluminescence resonance energy transfer (BRET(2)), we further suggest the existence of a physical interaction between D1R and D3R. Tagged D1R and D3R cotransfected in human embryonic kidney (HEK) 293 cells generated a significant BRET(2) signal that was insensitive to agonist stimulation, suggesting that they form a constitutive heterodimer. D1R and D3R regulate adenylyl cyclase (AC) in opposite ways. In HEK 293 cells coexpressing D1R and D3R, dopamine stimulated AC with higher potency and displaced [3H]R-(+)-7 chloro-8-hydroxy-3-methyl-1-phenyl-2,3,4,5-tetrahydro-1H-3-benzazepine (SCH23390) binding with higher affinity than in cells expressing the D1R. In HEK 293 cells individually expressing D1R or D3R, agonist stimulation induces internalization of D1R but not of D3R. Heterodimerization with D3R abolishes agonist-induced D1R cytoplasmic sequestration induced by selective D1R agonists and enables internalization of the D1R/D3R complex in response to the paired stimulation of both D1R and D3R. This mechanism involves beta-arrestin binding because it was blocked by mutant beta-arrestinV53D. These data suggest that as a result of dimerization, the D3R is switched to the desensitization mechanisms typical of the D1R. These data give a novel insight into how D1R and D3R may function in an integrated way, providing a molecular mechanism by which to converge D1R- and D3R related dysfunctions. PMID- 18424555 TI - PU.1 and C/EBPalpha/beta convert fibroblasts into macrophage-like cells. AB - Earlier work has shown that the transcription factor C/EBPalpha induced a transdifferentiation of committed lymphoid precursors into macrophages in a process requiring endogenous PU.1. Here we have examined the effects of PU.1 and C/EBPalpha on fibroblasts, a cell type distantly related to blood cells and akin to myoblasts, adipocytes, osteoblasts, and chondroblasts. The combination of the two factors, as well as PU.1 and C/EBPbeta, induced the up-regulation of macrophage/hematopoietic cell surface markers in a large proportion of NIH 3T3 cells. They also up-regulated these markers in mouse embryo- and adult skin derived fibroblasts. Based on cell morphology, activation of macrophage associated genes, and extinction of fibroblast-associated genes, cell lines containing an attenuated form of PU.1 and C/EBPalpha acquired a macrophage-like phenotype. The lines also display macrophage functions: They phagocytose small particles and bacteria, mount a partial inflammatory response, and exhibit strict CSF-1 dependence for growth. The myeloid conversion is primarily induced by PU.1, with C/EBPalpha acting as a modulator of macrophage-specific gene expression. Our data suggest that it might become possible to induce the transdifferentiation of skin-derived fibroblasts into cell types desirable for tissue regeneration. PMID- 18424556 TI - Deletion of G protein-coupled receptor 48 leads to ocular anterior segment dysgenesis (ASD) through down-regulation of Pitx2. AB - The development of the anterior segment of the mammalian eye is critical for normal ocular function, whereas abnormal development can cause glaucoma, a leading cause of blindness in the world. We report that orphan G protein-coupled receptor 48 (Gpr48/LGR4) plays an important role in the development of the anterior segment structure. Disruption of Gpr48 causes a wide spectrum of anterior segment dysgenesis (ASD), including microphthalmia, iris hypoplasia, irdiocorneal angle malformation, cornea dysgenesis, and cataract. Detailed analyses reveal that defective iris myogenesis and ocular extracellular matrix homeostasis are detected at early postnatal stages of eye development, whereas ganglion cell loss, inner nuclear layer thinness, and early onset of glaucoma were detected in 6-month-old Gpr48(-/-) mice. To determine the molecular mechanism of ASD caused by the deletion of Gpr48, we performed gene expression analyses and revealed dramatic down-regulation of Pitx2 in homozygous knockout mice. In vitro studies with the constitutively active Gpr48 mutant receptor demonstrate that Pitx2 is a direct target of the Gpr48-mediated cAMP-CREB signaling pathway in regulating anterior segment development, suggesting a role of Gpr48 as a potential therapeutic target of ASD. PMID- 18424557 TI - Arabidopsis TAO1 is a TIR-NB-LRR protein that contributes to disease resistance induced by the Pseudomonas syringae effector AvrB. AB - The type III effector protein encoded by avirulence gene B (AvrB) is delivered into plant cells by pathogenic strains of Pseudomonas syringae. There, it localizes to the plasma membrane and triggers immunity mediated by the Arabidopsis coiled-coil (CC)-nucleotide binding (NB)-leucine-rich repeat (LRR) disease resistance protein RPM1. The sequence unrelated type III effector avirulence protein encoded by avirulence gene Rpm1 (AvrRpm1) also activates RPM1. AvrB contributes to virulence after delivery from P. syringae in leaves of susceptible soybean plants, and AvrRpm1 does the same in Arabidopsis rpm1 plants. Conditional overexpression of AvrB in rpm1 plants results in leaf chlorosis. In a genetic screen for mutants that lack AvrB-dependent chlorosis in an rpm1 background, we isolated TAO1 (target of AvrB operation), which encodes a Toll-IL 1 receptor (TIR)-NB-LRR disease resistance protein. In rpm1 plants, TAO1 function results in the expression of the pathogenesis-related protein 1 (PR-1) gene, suggestive of a defense response. In RPM1 plants, TAO1 contributes to disease resistance in response to Pto (P. syringae pathovars tomato) DC3000(avrB), but not against Pto DC3000(avrRpm1). The tao1-5 mutant allele, a stop mutation in the LRR domain of TAO1, posttranscriptionally suppresses RPM1 accumulation. These data provide evidence of genetically separable disease resistance responses to AvrB and AvrRpm1 in Arabidopsis. AvrB activates both RPM1, a CC-NB-LRR protein, and TAO1, a TIR-NB-LRR protein. These NB-LRR proteins then act additively to generate a full disease resistance response to P. syringae expressing this type III effector. PMID- 18424558 TI - Small-molecule RETRA suppresses mutant p53-bearing cancer cells through a p73 dependent salvage pathway. AB - Identification of unique features of cancer cells is important for defining specific and efficient therapeutic targets. Mutant p53 is present in nearly half of all cancer cases, forming a promising target for pharmacological reactivation. In addition to being defective for the tumor-suppressor function, mutant p53 contributes to malignancy by blocking a p53 family member p73. Here, we describe a small-molecule RETRA that activates a set of p53-regulated genes and specifically suppresses mutant p53-bearing tumor cells in vitro and in mouse xenografts. Although the effect is strictly limited to the cells expressing mutant p53, it is abrogated by inhibition with RNAi to p73. Treatment of mutant p53-expressing cancer cells with RETRA results in a substantial increase in the expression level of p73, and a release of p73 from the blocking complex with mutant p53, which produces tumor-suppressor effects similar to the functional reactivation of p53. RETRA is active against tumor cells expressing a variety of p53 mutants and does not affect normal cells. The results validate the mutant p53 p73 complex as a promising and highly specific potential target for cancer therapy. PMID- 18424560 TI - Health-state utilities in liver disease: a systematic review. AB - OBJECTIVES: Health-state utilities are essential for cost-utility analysis. Few estimates exist for liver disease in the literature. The authors' aim was to conduct a systematic review of health-state utilities in liver disease, to look at the variation of study designs used, and to pool utilities for some liver disease states. METHODS: A search of MED-LINE, EMBASE, and CINAHL from 1966 to September 2006 was conducted including key words related to liver disease and utility measuring tools. Articles were included if health-state utility tools or expert opinion were used. Variance-weighted mean utility estimates were pooled using metaregression adjusting for disease state and utility assessment method. RESULTS: Thirty studies measured utilities of liver diseases/disease states. Half of these estimated utilities for hepatitis viruses: hepatitis A (n = 1), hepatitis B (n = 4), and hepatitis C (n = 10). Others included liver transplant (n= 6) and chronic liver disease (n= 5) populations. Twelve utility methods were used throughout. The EQ-5D (n = 10) was most popular method, followed by visual analogue scale (n = 9), time tradeoff (n = 6), and standard gamble (n = 4). Respondents were patients (n= 16), an expert panel (n = 10), non-liver diseases adults ( n=2), patient and expert (n = 1), and patient and healthy adult (n = 1). Type of perspective included community (n=21), patient (n=4), and both (n = 5). The pooled mean estimates in hepatitis C with moderate disease, compensated cirrhosis, decompensated cirrhosis, and post-liver transplant using the EQ-5D were 0.75, 0.75, 0.67, and 0.71, respectively. The change in these utilities using different methods were -0.07 (visual analogue scale), -0.01 (health utilities index version 3), +0.04 (standard gamble), + 0.08 (health utilities index version 2), + 0.12 (time tradeoff), and + 0.15 (standard gamble-transformed visual analogue scale). CONCLUSIONS: The authors have created a valuable liver disease- based utility resource from which researchers and policy makers can easily view all available utility estimates from the literature. They have also estimated health-state utilities for major states of hepatitis C. PMID- 18424559 TI - Impaired sodium excretion and increased blood pressure in mice with targeted deletion of renal epithelial insulin receptor. AB - Renal tubule epithelial cells express the insulin receptor (IR); however, their value has not been firmly established. We generated mice with renal epithelial cell-specific knockout of the IR by Cre-recombinase-loxP recombination using a kidney-specific (Ksp) cadherin promoter. KO mice expressed significantly lower levels of IR mRNA and protein in kidney cortex (49-56% of the WT) and medulla (32 47%) homogenates. Immunofluorescence showed the greatest relative reduction in the thick ascending limb and collecting duct cell types. Body weight, kidney weight, and food and water intakes were not different from WT littermates. However, KO mice had significantly increased basal systolic blood pressure (BP, 15 mm Hg higher) as measured by radiotelemetry. In response to a volume load by gavage (20 ml/kg of body weight, 0.9% NaCl, 15% dextrose), KO mice had impaired natriuresis (37 +/- 10 versus 99 +/- 9 mmol of Na(+) per 2 h in WT). Furthermore, volume load led to a sustained increase in BP in KO mice only. In contrast, insulin administration i.p. (0.5 units/kg of body weight) resulted in a significant fall in BP in WT, but not in KO mice. To test the role of reduced renal nitric oxide (NO) production in these responses, basal urinary nitrates plus nitrites excretion (UNOx) was measured and found to be 61% lower in KO vs. WT mice. Furthermore, acute insulin increased UNOx by 202% in the WT, relative to a significantly blunted rise (67%) in KO animals. These results illuminate a previously uncharacterized role for renal IR to reduce BP and facilitate sodium and water excretion, possibly via NO production. PMID- 18424561 TI - Infant pacifiers: an overview. PMID- 18424562 TI - An infant with crossed thumbs. PMID- 18424563 TI - Dipylidium caninum mimicking recurrent enterobius vermicularis (pinworm) infection. AB - Pinworm infection is a very common diagnosis in young children that is not always confirmed through laboratory evaluation before empiric therapy is prescribed. This article describes a toddler who was treated several times for pinworms because small white worms were seen in her perianal area. Laboratory analysis of parasite material found in her diaper later confirmed a diagnosis of dipylidiasis. Because the signs of dipylidiasis and pinworm infection overlap and the treatments for these parasitic infections are different, the laboratory should clinically confirm suspected persistent or recurrent pinworms. PMID- 18424564 TI - Unusual soft tissue manifestations of Neisseria meningitidis infections. AB - Soft tissue involvement is an unusual presenting feature for children with meningococcal infection. We describe 2 children, 1 with conjunctivitis and another with a thyroglossal duct cyst abscess associated with Neisseria meningitidis, and review previous reports of these entities to emphasize the broad spectrum of meningococcal disease and pertinent aspects of treatment and of prophylaxis of contacts. PMID- 18424565 TI - Failure to thrive. PMID- 18424566 TI - Rickets in an otherwise healthy 11-month-old. PMID- 18424567 TI - Domestic violence: its impact on children. PMID- 18424568 TI - Gastro-oesophageal reflux symptoms, oesophagitis and Barrett's oesophagus in the general population: the Loiano-Monghidoro study. AB - OBJECTIVE: Existing endoscopy-based data on gastro-oesophageal reflux disease (GORD) in the general population are scarce. This study aimed to evaluate typical symptoms and complications of GORD, and their associated risk factors, in a representative sample of the Italian population. METHODS: 1533 adults from two Italian villages were approached to undergo symptom assessment using a validated questionnaire and upper gastrointestinal endoscopy. Data were obtained from 1033 individuals (67.4% response rate). RESULTS: The prevalence of reflux symptoms was 44.3%; 23.7% of the population experienced such symptoms on at least 2 days per week (frequent symptoms). The prevalence rates of oesophagitis and Barrett's oesophagus in the population were 11.8% and 1.3%, respectively. Both frequent (relative risk (RR) 2.6; 95% confidence interval (CI) 1.7 to 3.9) and infrequent (RR 1.9; 95% CI 1.2 to 3.0) reflux symptoms were associated with the presence of oesophagitis. No reflux symptoms were reported by 32.8% of individuals with oesophagitis and 46.2% of those with Barrett's oesophagus. Hiatus hernia was associated with frequent reflux symptoms and oesophagitis, and was present in 76.9% of those with Barrett's oesophagus. We found no association between body mass index and reflux symptoms or oesophagitis. CONCLUSIONS: GORD is common in Italy, but the prevalence of Barrett's oesophagus in the community is lower than has been reported in selected populations. Both frequent and infrequent reflux symptoms are associated with an increased risk of oesophagitis. Individuals with oesophagitis and Barrett's oesophagus often have no reflux symptoms. PMID- 18424569 TI - The development and validation of the Khon Kaen University Pediatric Pain Assessment Tool for school-aged Isaan children in Thailand. AB - This research was aimed at developing a tool for pain assessment specifically for parents to use with children of the Isaan culture of Northeast Thailand. The Khon Kaen University (KKU) Pediatric Pain Assessment Tool was first constructed through the use of a qualitative study and literature reviews. Then, it was modified using the results of a pilot study with 38 stakeholders. Seventeen nurses, 150 postoperative children, and 150 family caregivers participated in a study comparing the tool with the standard Faces Scale and Numeric Rating Scale. Results revealed no difference in children's pain scores, as well as no statistical difference in children's pain score ratings between the three groups, suggesting the new tool is appropriate for Isaan children. PMID- 18424570 TI - Gastric cancer and Helicobacter pylori: the bug, the host or the environment? PMID- 18424571 TI - Prescribing errors by family practice residents. PMID- 18424572 TI - Current and future management of chronic hepatitis C infection. AB - Current treatment for patients with chronic hepatitis C virus (HCV) infection consists of the combination of pegylated interferon and ribavirin. This treatment regimen achieves a sustained virological response, defined as undetectable HCV RNA 6 months after treatment cessation, in 50% of patients overall. There is therefore a need for new treatments to improve the sustained virological response rate and reduce the number of adverse effects associated with pegylated interferon and ribavirin. This review examines the current management of chronic HCV infection, including who is eligible for treatment, the optimum duration of treatment, and management of side effects. New drugs in development, such as HCV specific protease inhibitors, polymerase inhibitors, immune modulators and ribavirin analogues, are outlined, and their role in the treatment armamentarium is discussed, whether used alone or in combination with existing treatments. PMID- 18424573 TI - Relevance of clotting tests in liver disease. AB - Liver disease is associated with impairment of the haemostatic function due to the abnormal and decreased synthesis of the clotting factors. It is thus only logical to have considered assessment of the clotting profile (to include prothrombin time (PT) and activated partial thromboplastin time (aPTT)) to be an integral part of the comprehensive assessment of a patient who presents with liver impairment. Laboratory abnormalities of coagulation are considered to be a predictive risk factor for bleeding, but patients with liver disease do not have bleeding pattern as those who have coagulation factor deficiencies. Recent experiments have cast doubts over the use of PT and aPTT as a marker of bleeding in liver disease and the use of such tests to decide the need for plasma replacement before interventions like liver biopsy. This article reviews the relevance of the clotting profile in liver disease, the other factors involved in the haemostatic failure associated with it, and the technical problems in the interpretation of these results. Most importantly, it stresses the need for more trials to help us guide the management of bleeding in patients with liver impairment. PMID- 18424575 TI - Diabetic heart disease. AB - Diabetes mellitus is responsible for a spectrum of cardiovascular disease. The best known complications arise from endothelial dysfunction, oxidation, inflammation, and vascular remodelling and contribute to atherogenesis. However, the effects on the heart also relate to concurrent hypertensive heart disease, as well as direct effects of diabetes on the myocardium. Diabetic heart disease, defined as myocardial disease in patients with diabetes that cannot be ascribed to hypertension, coronary artery disease, or other known cardiac disease, is reviewed. PMID- 18424576 TI - Prognostic significance of genotyping Helicobacter pylori infection in patients in younger age groups with gastric cancer. AB - BACKGROUND: Malignant tumours of the stomach are common, but the incidence of stomach cancer varies from country to country, probably a result of genetic, epigenetic and environmental factors. Stomach cancer often occurs in older people whose stomachs produce only small quantities of acid. Although infection with Helicobacter pylori has been proven beyond doubt in the aetiopathogenesis of various gastric disorders, not much is known about the genotypes of H pylori infection in early-onset gastric cancer. AIM: To ascertain the genotypes of H pylori in gastric cancer. METHODS: Ninety-two patients were separated into three groups on the basis of their endoscopic findings: group 1, gastric cancer; group 2, gastric ulcer; group 3, non-ulcer dyspepsia. Gastric biopsy specimens were obtained for culture and DNA isolation; additional specimens were taken from subjects with gastric cancer for histopathological analysis. Amplification was performed using specific oligonucleotide primers to obtain genotypic data. Four samples from each group were randomly selected for sequence analysis. RESULTS: Genotypic analysis showed cagT+ve/hrgA+ve/cagA+ve/cagE+ve/vacAs1+ve to be highly prevalent in 79% of cases of H pylori infection. This genotype was found in 88% of subjects in group 1 and 78% in group 2. Intestinal-type adenocarcinoma was found in 35 subjects (83%), 32 (9%) of which harboured this genotype. Sequence analysis showed no significant strain-specific variations. CONCLUSIONS: Certain genotypes of H pylori have higher predictive value for the development of intestinal-type carcinoma at an early age. Genotyping of H pylori may well be a useful tool for screening people at increased risk of developing malignancy. PMID- 18424577 TI - Prescription writing skills of residents in a family practice residency programme in Bahrain. AB - PURPOSE OF THE STUDY: To evaluate the prescription writing skill of final year residents in a family practice residency programme (FPRP) in Bahrain, and to compare skill of residents who have graduated from medical schools with problem based learning (PBL) versus traditional (non-PBL) curricula. STUDY DESIGN: Prescriptions issued by the residents were prospectively collected for two consecutive cohorts in May 2004 and May 2005. Prescription errors were classified as errors of omission (minor and major), commission (incorrect information) and integration (drug-drug interactions). RESULTS: In 69.6% of medications with major omission errors, dosage form (39.4%) and length of treatment (18.5%) were not specified. In 24.7% of medications with commission errors, dosing frequency (19.9%) and incorrect strength/dose (2.2%) were the most common errors. Integration errors comprised 5.7% of all prescribing errors. No significant differences were observed between PBL and non-PBL graduates with regard to the total number of prescriptions with errors, drugs per prescription, polypharmacy, and the total number of drugs with errors. The proportion of prescriptions with a potential for drug-drug interactions was comparable between PBL and non-PBL graduates. PBL graduates prescribed medications using brand names at a rate greater than non-PBL, whereas non-PBL graduates prescribed medications on inappropriate "as required" basis, and injections at a rate greater than PBL residents. CONCLUSIONS: Prescription writing skill of the final year residents in an FPRP programme was suboptimal for both PBL and non-PBL graduates. Integration of prescription writing skill and a rational pharmacotherapeutic programme into the FPRP curriculum is recommended. PMID- 18424578 TI - Cardiac autonomic neuropathy in diabetes mellitus: prevalence, risk factors and utility of corrected QT interval in the ECG for its diagnosis. AB - OBJECTIVES: To study the prevalence and risk factors for cardiac autonomic neuropathy (CAN) and the utility of prolongation of corrected QT interval (QTc) in the ECG to diagnose CAN in patients with diabetes mellitus. DESIGN AND SETTING: Cross-sectional study conducted among patients attending the diabetic clinic of a teaching hospital. METHODS: The prevalence of CAN among 100 patients with type 1 and type 2 diabetes mellitus was assessed by the five autonomic function tests by Eving's methodology. The CAN score in each patient and its relationship to the QTc interval were analysed. Possible influences of age, duration of diabetes and coexistent peripheral neuropathy on the occurrence of CAN also were studied. RESULTS: The prevalence of CAN was 60%. Univariate analysis showed a significant association between CAN and higher age (odds ratio (OR) 15.75), prolongation of QTc (OR 5.55), duration of disease over 10 years (OR 2) and peripheral neuropathy (p<0.001) in patients with type 1 diabetes. Significant risks for CAN among patients with type 2 diabetes were coexistent peripheral neuropathy (OR 14), prolonged QTc (OR 9.75), higher age (OR 7.2) and disease duration over 10 years (OR 1.92) in univariate analysis, but none of them showed independent risk in multivariate analysis. Disease duration over 10 years resulted in QTc prolongation in a significant numbers of cases with type 1 (p<0.001) and type 2 (p = 0.006) diabetes. The sensitivity, specificity and positive predictive value of QTc prolongation for the diagnosis of CAN were 77%, 62.5% and 77% in type 1 and 76.5%, 75% and 81.3% in type 2, respectively. Higher CAN scores correlated with longer QTc intervals (coefficient of correlation 0.73; p<0.001). CONCLUSIONS: The prevalence of CAN in diabetes mellitus is high. Higher age, longer duration of diabetes and peripheral neuropathy are significant risk factors. QTc interval in the ECG can be used to diagnose CAN with reasonable sensitivity, specificity and positive predictive value. PMID- 18424579 TI - Educational quality improvement report: outcomes from a revised morbidity and mortality format that emphasised patient safety. AB - PROBLEM: Although Morbidity and Mortality conferences (MMC) were originally designed to promote quality care through careful analysis of adverse events, focus on individual actions or the fear of incrimination may interfere with identification of system issues contributing to the adverse outcomes. DESIGN: Pre and post-intervention assessments of participant attitudes toward patient safety and conference redesign were performed utilizing an attitudinal survey. Participants provided a unique identifier for paired-means procedure. A list of contributing factors, recommended solutions, and targeted system improvements was maintained with ongoing progress recorded. SETTING: Department of Internal Medicine training program at University of Missouri-Columbia, an academic health care center affiliated with the University of Missouri Hospitals and Clinics and the Harry S. Truman Veteran's Administration Hospital. PARTICIPANTS: Residents and fellows from the Department of Internal Medicine residency program. EDUCATIONAL OBJECTIVES: (1) Distinguish between culture of blame/shame and patient safety culture, (2) Identify gaps in quality contributing to adverse outcomes, (3) Identify strategies to close gaps, (4) Participate in root cause analysis, demonstrating an ability to review an adverse event and recommend an action plan. STRATEGIES FOR CHANGE: An interdisciplinary team modified the internal medicine MMC to emphasize a better understanding of patient safety principles and system-based practice interventions. For each adverse event analyzed, root causes were identified, followed by discussion of system interventions that might prevent future such events. KEY MEASURES FOR IMPROVEMENT: (1) Attitudes of residents and fellows regarding patient safety, as measured on a 20 item, five-point ordinal scale (strongly disagree to strongly agree) survey, (2) System improvements generated from the Patient Safety M&M Conferences (PSMMC), and (3) Attendance at PSMMC. EFFECTS OF CHANGE: Clinical outcomes: Conference participants offered 121 system improvement recommendations; 39 suggested system interventions were pursued based upon the likelihood of achieving high impact changes. These targeted changes were assigned to department/facility representatives with 23 (59%) improvements implemented, 11 (28%) partially implemented or in progress, and five (13%) abandoned due to impracticality or redundancy. Educational outcomes: Surveys were completed by 58 residents and fellows before and after modification of conference format. Six of the 20 survey items showed significant change with four of these changes occurring in the desired direction. Eleven of the remaining 14 responses changed in the desired direction, but did not reach statistical significance. Average MMC attendance increased from 41+/-8 to 50+/-10 (p<0.03) participants. LESSONS LEARNT: The new PSMMC initiated multiple improvements in the quality of patient care without sacrificing attendance or attitudes of the residents or fellows. The new PSMMC promotes opportunities for participants to improve quality of patient care in a safe and nurturing environment. PMID- 18424580 TI - Drug interaction with anti-mycobacterial treatment as a cause of clopidogrel resistance. AB - The case is reported of a patient presenting with recurrent acute myocardial infarction due to stent thrombosis resulting from possible clopidogrel resistance caused by interaction between clopidogrel and anti-tuberculosis drugs. A brief overview of drug interaction as a risk factor for clopidogrel resistance is given. PMID- 18424581 TI - Importance of checking anti-glomerular basement membrane antibody status in patients with anti-neutrophil cytoplasmic antibody-positive vasculitis. AB - The case is reported of a 68-year-old man with perinuclear anti-neutrophil cytoplasmic antibody (pANCA)-associated glomerulonephritis who developed antibodies to glomerular basement membrane (anti-GBM) resulting in end stage renal failure. His pANCA titre on admission was 1:1024 IgG and he was anti myeloperoxidase positive. A renal biopsy showed advanced sclerosing necrotising glomerulonephritis consistent with a pauci-immune ANCA-positive glomerulonephritis. He was treated with steroids and cyclophosphamide. His serum creatinine profile improved. He had a relapse of disease 16 months later, which was successfully treated. After a further 16 months, he presented with acute renal failure (creatinine 1060 micromol/l). His pANCA titre on admission was 1:64 IgG. This was treated as a further relapse of ANCA-positive vasculitis. He became oliguric and his haemoglobin concentration fell. Eight days after admission, he was found to be strongly positive for anti-GBM (138 U/ml). Despite receiving cyclophosphamide, steroids and plasma exchange, he remained dialysis-dependent. PMID- 18424582 TI - A day out with Darwin. PMID- 18424584 TI - Surveillance of work-related musculoskeletal disorders. PMID- 18424583 TI - Growing knowledge about "what works" to prevent work injuries. PMID- 18424586 TI - James Allen Olson (1924-2000). PMID- 18424587 TI - Harriette Chick and the problem of rickets. PMID- 18424588 TI - Cinnamon polyphenol extract affects immune responses by regulating anti- and proinflammatory and glucose transporter gene expression in mouse macrophages. AB - Tristetraprolin (TTP/zinc finger protein 36) family proteins have antiinflammatory effects by destabilizing proinflammatory mRNA. TTP expression is reduced in fats of obese people with metabolic syndrome and brains of suicide victims and is induced by insulin and cinnamon polyphenol extract (CPE) in adipocytes, by lipopolysaccharide (LPS) in macrophages, and by green tea polyphenol extract in rats. CPE was reported to improve immune function against microorganisms, but the mechanism is unknown. This study tested the hypothesis that CPE regulates immune function involving genes encoding TTP, proinflammatory cytokines, and glucose transporter (GLUT) families and compared the effects of CPE to those of insulin and LPS in mouse RAW264.7 macrophages. CPE increased TTP mRNA and protein levels, but its effects were less than LPS. CPE (100 mg/L, 0.5-4 h) increased TTP and tumor necrosis factor (TNF) mRNA levels by up to 2- and 6 fold that of the control, respectively, and the base level of TTP was 6-fold that of TNF. LPS (0.1 mg/L, 4 h) increased TTP, TNF, granulocyte-macrophage colony stimulating factor, cyclooxgenase-2, and interleukin 6 mRNA levels by 39-1868 fold. CPE and LPS increased GLUT1 expression (the major GLUT form in macrophages) to 3- and 2-fold that of the control, respectively. Insulin (100 nmol/L, 0.5-4 h) did not exhibit major effects on the expression of these genes. CPE increased TTP expression more rapidly than those of proinflammatory cytokines and the net increases of TTP mRNA levels were larger than those of proinflammatory cytokines. These results suggest that CPE can affect immune responses by regulating anti- and proinflammatory and GLUT gene expression. PMID- 18424590 TI - Copper deficiency increases the virulence of amyocarditic and myocarditic strains of coxsackievirus B3 in mice. AB - Deficiency in several trace elements, including copper and selenium, is associated with increased levels of oxidative stress. Copper deficiency also has been shown to impair immune function. Previous work by others demonstrated that passage of an amyocarditic or myocarditic strain of coxsackievirus B3 (CVB3) through selenium- or vitamin E-deficient mice led to increased cardiac pathology. To determine whether a copper deficiency would similarly alter the pathogenesis of CVB3 infections, Swiss outbred dams and their litters were fed copper deficient diets from birth and received either deionized water or water with 0.315 mmol/L copper as copper sulfate. At 4 wk of age, copper-adequate or deficient male and female offspring were infected with an amyocarditic or myocarditic strain of CVB3. Heart titers were elevated at d 3 and 7 postinfection in copper-deficient mice infected with the myocarditic CVB3 strain (CVB3/20) but only at d 7 in deficient mice infected with the amyocarditic CVB3 strain (CVB3/0) compared with copper-adequate controls. Copper-deficient mice infected with either strain of CVB3 had increased cardiac pathology compared with copper adequate controls. Genomic sequences of viruses isolated from copper-adequate and -deficient mice were identical. Heart cytokine expression was elevated in copper deficient CVB3-infected mice compared with infected controls. Circulating CVB3 specific IgG2a but not IgM levels were decreased in copper-deficient mice. Thus, copper deficiency is associated with an increased inflammatory response but decreased acquired immune response to CVB3 infection that results in increased cardiac pathology, presumably due to increased viral load. PMID- 18424591 TI - Energy restriction results in a mass-adjusted decrease in energy expenditure in cats that is maintained after weight regain. AB - Dietary energy restriction (ER) is used to treat obesity in cats but it is often unsuccessful. The purpose of this study was to determine whether ER results in a sustained decrease in mass-adjusted energy expenditure (EE) that may oppose weight loss and promote weight regain. EE and body composition were measured in 10 adult neutered cats at 3 time points: baseline (obese cats), during weight loss (40% ER), and following weight regain. The cats started with a body weight (BW) of 6.1 +/- 0.30 kg, body condition score (BCS) of 7.6 +/- 0.14 (on a 9-point scale), and fat body mass (FM) of 38 +/- 1.0% of BW. After weight loss, BW was 5.0 +/- 0.19 kg, BCS was 5.5 +/- 0.07 kg, and FM was 31 +/- 1.6% (P < 0.01). After weight regain, BW was 6.2 +/- 0.30 kg, BCS was 7.7 +/- 0.16, and FM was 42 +/- 1.8% (P < 0.01). Total EE decreased from 1258 +/- 33.7 kJ/d to 1025 +/- 39.6 kJ/d during weight loss (P < 0.001). After weight regain, EE was still lower than baseline (1103 +/- 41.5 kJ/d, P < 0.001). Energy intake (EI) at baseline (1337 +/ 50.6 kJ/d) was higher than EI after weight loss and regain (1217 +/- 61.2 kJ/d), resulting in no differences in energy balance (78 +/- 30.4 and 104 +/- 35.4 kJ/d, respectively, P = 0.581). These results support the hypothesis that ER results in a mass-adjusted decrease in EE in cats that is maintained after weight regain. PMID- 18424589 TI - Gamma-synuclein is an adipocyte-neuron gene coordinately expressed with leptin and increased in human obesity. AB - Recently, we characterized tumor suppressor candidate 5 (Tusc5) as an adipocyte neuron PPARgamma target gene. Our objective herein was to identify additional genes that display distinctly high expression in fat and neurons, because such a pattern could signal previously uncharacterized functional pathways shared in these disparate tissues. gamma-Synuclein, a marker of peripheral and select central nervous system neurons, was strongly expressed in white adipose tissue (WAT) and peripheral nervous system ganglia using bioinformatics and quantitative PCR approaches. Gamma-synuclein expression was determined during adipogenesis and in subcutaneous (SC) and visceral adipose tissue (VAT) from obese and nonobese humans. Gamma-synuclein mRNA increased from trace levels in preadipocytes to high levels in mature 3T3-L1 adipocytes and decreased approximately 50% following treatment with the PPARgamma agonist GW1929 (P < 0.01). Because gamma-synuclein limits growth arrest and is implicated in cancer progression in nonadipocytes, we suspected that expression would be increased in situations where WAT plasticity/adipocyte turnover are engaged. Consistent with this postulate, human WAT gamma-synuclein mRNA levels consistently increased in obesity and were higher in SC than in VAT; i.e. they increased approximately 1.7-fold in obese Pima Indian adipocytes (P = 0.003) and approximately 2-fold in SC and VAT of other obese cohorts relative to nonobese subjects. Expression correlated with leptin transcript levels in human SC and VAT (r = 0.887; P < 0.0001; n = 44). Gamma synuclein protein was observed in rodent and human WAT but not in negative control liver. These results are consistent with the hypothesis that gamma synuclein plays an important role in adipocyte physiology. PMID- 18424592 TI - Restoration of body energy reserves during refeeding in rats is dependent on both the intensity of energy restriction and the metabolic status at the onset of refeeding [corrected]. AB - During starvation, after a short dynamic period of adaptation (phase I), a metabolic steady state is reached in which proteins are spared and lipids provide most of the energy expended [phase II (P2)]. However, protein breakdown increases dramatically once a lower threshold of body lipids is reached [phase III (P3)]. Body composition, energy intake, energy expenditure, and energy efficiency were determined in 8 groups of rats (fed, food-deprived up to P2 or P3 of starvation and refed for 3 d, 7 d, or until body mass restoration) to determine whether the kinetics of lipid and/or protein reserve recovery may be slowed down when refeeding occurs after the lipid threshold has been reached. Despite larger losses, P3 refed rats restored their body reserves as efficiently as those refed in P2. Whatever the nutritional status at the onset of refeeding, rehydration occurred first and hyperphagia played a more important role than hypometabolism in the restoration of the lost reserves. However, the pattern of body component gains was different during early refeeding. In P3 refed rats, body lipids were restored preferentially by significant contribution from endogenous lipid production. Thus, the extent of lipid depletion has important consequences for the restoration pattern of the body reserves. It depends not only on the intensity of the energy restriction (partial or total) as already demonstrated but also on the metabolic status at the onset of refeeding. These results may have significant implications on the way refeeding should be conducted after severe energy depletion. PMID- 18424593 TI - Dietary arginine supplementation increases mTOR signaling activity in skeletal muscle of neonatal pigs. AB - Dietary arginine supplementation increases growth of neonatal pigs, but the underlying mechanisms are unknown. This study was conducted to test the hypothesis that the arginine treatment activates translation initiation factors and protein synthesis in skeletal muscle. Piglets were fed milk-based diets supplemented with 0 or 0.6% L-arginine between 7 and 14 d of age. Following a 7-d period of arginine supplementation, at 1 h after the last meal, jugular venous blood samples were obtained for metabolite analysis, whereas longissimus muscle and liver were collected to determine the abundance and phosphorylation state of the mammalian target of the rapamycin (mTOR), ribosomal protein S6 kinase 1 (S6K1), eukaryotic initiation factor (eIF) 4E-binding protein-1 (4E-BP1), eIF4E, and eIF4G. Fractional rates of protein synthesis were measured in muscle and liver using the [(3)H]phenylalanine flooding-dose technique. Arginine supplementation increased (P < 0.05) daily gain, the plasma insulin concentration, and protein synthesis in skeletal muscle but not in liver. The arginine treatment enhanced the formation of the active eIF4E x eIF4G complex but reduced the amount of the inactive 4E-BP1 x eIF4E complex in muscle. These changes were associated with elevated levels of phosphorylated mTOR and 4E-BP1 in muscle of arginine-supplemented piglets (P < 0.05). Neither the total amounts nor phosphorylation levels of the translation initiation factors in the liver differed between control and arginine-supplemented piglets. Collectively, these results suggest that dietary arginine supplementation increases mTOR signaling activity in skeletal muscle, but not in liver, of milk-fed neonatal pigs. The findings provide a molecular mechanism for explaining the previous observation that increased circulating arginine stimulated muscle protein synthesis and promoted weight gain in neonatal pigs. PMID- 18424594 TI - L-alpha-glycerophosphocholine contributes to meat's enhancement of nonheme iron absorption. AB - In this research, our aim was to isolate and characterize the substance known as "meat factor," which is reported to enhance nonheme iron absorption. We used various analytical techniques, and the final step was a human study to measure the effect of a candidate compound on iron absorption. Lean beef was selected for study, as it is known to increase nonheme iron absorption. Cooked ground beef was homogenized and aliquots were taken through a simulated gastric and intestinal digestion. This was followed by purification using fast protein liquid chromatography. The fractions were collected and applied to a Caco-2 cell system designed to measure iron absorption using radioiron. Fractions with an enhancing effect were analyzed by mass spectrometry, nuclear magnetic resonance, and HPLC, and a proposed empirical formula was obtained for the substance in the most active fraction (C(8)H(20) NO(6)P). Tandem mass spectrometry was used to identify the compound as L-alpha-glycerophosphocholine (L-alpha) by comparing the spectra against authentic material. We added a commercially available food grade source of L-alpha to vegetarian lasagna, with and without 100 mg ascorbic acid (a known enhancer of nonheme iron absorption), at the same enhancer:iron molar ratio (2:1), and fed meals to 13 women of child-bearing age with low iron stores. The nonheme iron was labeled with stable isotopes of iron to provide a total dose per meal of 10 mg iron, and absorption was measured from erythrocyte incorporation. Nonheme iron absorption from lasagna was increased by the addition of either ascorbic acid (P = 0.010) or L-alpha (P = 0.023). We have identified L-alpha as a component of muscle tissue that enhances nonheme iron absorption, and this finding provides new opportunities for iron fortification of foods. PMID- 18424595 TI - Ferritin-iron is released during boiling and in vitro gastric digestion. AB - Biofortification of staple foods with iron in the form of ferritin-iron is a promising approach to fighting iron-deficiency anemia in developing countries. However, contradictory results regarding iron bioavailability to humans from ferritin are not yet fully clarified. Furthermore, the question has been raised whether ferritin can potentially survive gastric passage intact and be absorbed via a ferritin-specific uptake mechanism. We studied changes of ferritin-iron and protein during cooking and in vitro gastric digestion. Water soluble, native ferritin-iron, measured in different legumes, represented 18% (soybeans) up to maximally 42% (peas) of total seed iron. Ferritin-iron was no longer detectable after boiling the legumes for 50 min in excess water. When the same cooking treatment was applied to recombinant bean ferritin propagated in Escherichia coli, some ferritin-iron remained measurable. During in vitro gastric digestion of recombinant bean ferritin and red kidney bean extract, ferritin-iron was fully released from the protein and dissolved at pH 2. Stability tests at varying pH at 37 degrees C showed that the release of ferritin-iron starts at pH 5 and is complete at pH 2. We concluded that ferritin-iron is efficiently released from the ferritin molecule during cooking and at gastric pH and that it should be absorbed as efficiently as all other nonheme iron in food. PMID- 18424596 TI - Quercetin from shallots (Allium cepa L. var. aggregatum) is more bioavailable than its glucosides. AB - The lipophilic character of quercetin suggests that it can cross enterocyte membranes via simple diffusion. Therefore, it should be more bioavailable than its glucosides, which require preliminary hydrolysis or active transport for absorption. However, the published human studies show that quercetin is less bioavailable than its glucosides. Assuming that low bioavailability of quercetin aglycone provided to humans as a pure substance is the result of its low solubility in the digestive tract, we studied its bioavailability from dietary sources in which quercetin was dispersed in the food matrix. In a randomized crossover study, 9 volunteers took a single dose of either shallot flesh (99.2% quercetin glucosides and 0.8% quercetin aglycone) or dry shallot skin (83.3% quercetin aglycone and 16.7% quercetin glucosides), providing 1.4 mg quercetin per kg of body weight. Blood samples were collected before and after consumption of shallot preparations. Plasma quercetin was measured on HPLC with electrochemical detection after plasma enzymatic treatment. The maximum plasma quercetin concentration of 1.02 +/- 0.13 micromol/L was reached at 2.33 +/- 0.50 h after shallot flesh consumption compared with 3.95 +/- 0.62 micromol/L at 2.78 +/- 0.15 h after dry skin consumption. The area under the concentration-time curve after dry skin consumption was 47.23 +/- 7.53 micromol x h(-1) x L(-1) and was significantly higher than that after shallot flesh intake (22.23 +/- 2.32 micromol x h(-1) x L(-1)). When provided along with dietary sources, quercetin aglycone is more bioavailable than its glucosides in humans. Results point to the food matrix as a key factor. PMID- 18424597 TI - Fish oil increases the duodenal flow of long chain polyunsaturated fatty acids and trans-11 18:1 and decreases 18:0 in steers via changes in the rumen bacterial community. AB - Ruminant fat is rich in SFA, partly due to the biohydrogenation of dietary PUFA to SFA in the rumen. This process can be inhibited by the dietary inclusion of fish oil. The only bacteria isolated from the rumen capable of converting PUFA to SFA are closely related to Clostridium proteoclasticum. The aim of this study was to investigate if a correlation could be found in vivo between dietary fish oil inclusions and the composition of the ruminal bacterial community and specifically of C. proteoclasticum. Six Hereford x Friesian steers, prepared with ruminal and duodenal cannulae, received grass silage plus 1 of 3 concentrates resulting in total dietary fish oil contents of 0, 1, or 3% of dry matter. A dual flow marker technique was employed to estimate the relative flow of fatty acids. Steers fed the 3% fish oil diet had 100% increases in trans 18:1 flow, whereas 18:0 flow declined to 39% of steers fed the control diet. 16S ribosomal RNA-based denaturing gradient gel electrophoresis profiles obtained from ruminal digesta showed major changes in the bacterial community within steers fed the 3% fish oil diet. Quantitative PCR indicated only a weak relation between numbers of C. proteoclasticum and 18:0 flow between treatments and in individual steers (P < 0.05, but the percentage variance accounted for only 22.8) and did not provide unambiguous evidence that numbers of C. proteoclasticum in the rumen dictate the ratios of SFA:PUFA available for absorption by the animal. Understanding which microbes biohydrogenate PUFA in the rumen is key to developing novel strategies to improve the quality of ruminant products. PMID- 18424598 TI - Anthocyanin excretion by humans increases linearly with increasing strawberry dose. AB - A clinical study was conducted to investigate the dose response and metabolism of strawberry anthocyanins. In a crossover study design, 12 healthy adults consumed each of 3 strawberry treatments. The treatments were 100 g, 200 g, and 400 g of pureed strawberries, delivering 15 micromol, 30 micromol, and 60 micromol anthocyanin, respectively. Urine samples were collected for 24 h after each dose and samples were analyzed by HPLC with diode array detection and ion trap MS. Pelargonidin 3-glucoside was the major anthocyanin form in the treatments, and pelargonidin 3-glucoside and 3 metabolites of pelargonidin 3-glucoside (detected as monoglucuronides) were excreted in urine after ingestion. One predominant monoglucuronide form was detected in urine in masses 10-fold higher than the other 2 monoglucuronide forms. Increasing dose resulted in increasing appearance of anthocyanins in urine, and the mass of each pelargonidin monoglucuronide increased in urine with increasing dose. These results suggest that pelargonidin 3-glucoside absorption and metabolism are not saturated at masses < or = 60 micromol, thus showing that more strawberry anthocyanin can be absorbed with increasing dose. PMID- 18424599 TI - PPARgamma mRNA expression is reduced in peripheral blood mononuclear cells after fat overload in patients with metabolic syndrome. AB - PPARgamma is a transcriptional regulator of metabolism; its activity can be modulated by direct binding of dietary lipids. The most prevalent human PPARgamma gene variant, Ala12, is associated with postprandial hypertriglyceridemia in patients with metabolic syndrome, although the mechanism whereby this polymorphism affects lipid homeostasis remains to be fully determined. Using peripheral blood mononuclear cells (PBMC), we studied the effect of the Pro12 and Ala12 polymorphisms on mRNA expression of PPARgamma and nuclear factor kappa B genes before and 3 and 4 h after fat overload. We also studied several biochemical and oxidative stress variables. Most of the indicators of oxidative stress were higher in patients with metabolic syndrome than in healthy subjects before and after fat overload. Patients also differed depending on whether they had the Pro12 or Ala12 variant in PBMC; PPARgamma expression was lower in healthy subjects compared with patients. After fat overload, circulating triglycerides and PPARgamma expression were positively correlated (r = 0.617, P < 0.05), and PPARgamma expression tended to be negatively correlated with 2 important markers of oxidative stress: plasma lipid peroxidation (r = -0.224, P < 0.1) and carbonylated proteins (CPro) (r = -0.340, P < 0.1) concentrations. We also found differences in several indicators of oxidative stress between Pro12 and Ala12 patients, including an increase in plasma CPro before and after fat overload in Ala12 but not Pro12 patients. These data provide evidence that the Ala12 sequence variant is associated with a worse metabolic profile than Pro12. This is related to differences in the expression of PPARgamma and to oxidative imbalance after fat overload. PMID- 18424600 TI - Almonds reduce biomarkers of lipid peroxidation in older hyperlipidemic subjects. AB - Nut consumption has been associated with reduced coronary heart disease (CHD) risk. In addition to cholesterol-lowering properties, almonds have been shown to lower oxidized LDL concentrations. However, little is known regarding their effects on other markers of oxidative stress. The dose-response effects of whole almonds, taken as snacks, were compared with low-saturated fat (<5% energy) whole wheat muffins (control) in the therapeutic diets of hyperlipidemic subjects. In a randomized crossover study, 27 hyperlipidemic men and women consumed 3 isoenergetic (mean 423 kcal/d or 1770 kJ/d) supplements each for 1 mo. Supplements consisted of full-dose almonds (73 +/- 3 g/d), half-dose almonds plus half-dose muffins (half-dose almonds), and full-dose muffins (control). Subjects were assessed at wk 0, 2 and 4. Mean body weights differed < or = 300 g between treatments, although the weight loss on the half-dose almond treatment was greater than on the control (P < 0.01). At 4 wk, the full-dose almonds reduced serum concentrations of malondialdehyde (MDA) (P = 0.040) and creatinine-adjusted urinary isoprostane output (P = 0.026) compared with the control. Serum concentrations of alpha- or gamma-tocopherol, adjusted or unadjusted for total cholesterol, were not affected by the treatments. Almond antioxidant activity was demonstrated by their effect on 2 biomarkers of lipid peroxidation, serum MDA and urinary isoprostanes, and supports the previous finding that almonds reduced oxidation of LDL-C. Antioxidant activity provides an additional possible mechanism, in addition to lowering cholesterol, that may account for the reduction in CHD risk with nut consumption. PMID- 18424601 TI - Divergent associations of plasma choline and betaine with components of metabolic syndrome in middle age and elderly men and women. AB - Choline is involved in the synthesis of phospholipids, including blood lipids, and is the immediate precursor of betaine, which serves as a methyl group donor in a reaction converting homocysteine to methionine. Several cardiovascular risk factors are associated with plasma homocysteine, whereas little is known about their relationship to choline and betaine. We examined the relation of plasma choline and betaine to smoking, physical activity, BMI, percent body fat, waist circumference, blood pressure, serum lipids, and glucose in a population-based study of 7074 men and women aged 47-49 and 71-74 y. Overall plasma concentrations (means +/- SD) were 9.9 +/- 2.3 micromol/L for choline and 39.5 +/- 12.5 micromol/L for betaine. Choline and betaine were lower in women than in men and in younger subjects compared with older (P < 0.0001). Multivariate analyses showed that choline was positively associated with serum triglycerides, glucose, BMI, percent body fat, waist circumference (P < 0.0001 for all), and physical activity (P < 0.05) and inversely related to HDL cholesterol (P < 0.05) and smoking (P < 0.0001). Betaine was inversely associated with serum non-HDL cholesterol, triglycerides, BMI, percent body fat, waist circumference, systolic and diastolic blood pressure (P < 0.0001 for all), and smoking (P < 0.05) and positively associated with HDL cholesterol (P < 0.01) and physical activity (P < 0.0001). Thus, an unfavorable cardiovascular risk factor profile was associated with high choline and low betaine concentrations. Choline and betaine were associated in opposite directions with key components of metabolic syndrome, suggesting a disruption of mitochondrial choline dehydrogenase pathway. PMID- 18424602 TI - Lysozyme transgenic goats' milk influences gastrointestinal morphology in young pigs. AB - Transgenesis provides a method of expressing novel proteins in milk to increase the functional benefits of milk consumption. Transgenic goats expressing human lysozyme (hLZ) at 67% of the concentration in human breast milk were produced, thereby enhancing the antimicrobial properties of goats' milk. The objective of this study was to investigate the impact of pasteurized milk containing hLZ on growth, the intestinal epithelium, and an enteropathogenic Escherichia coli (EPEC) infection in young weaned pigs. Pigs were placed into 4 groups and fed a diet of solid food and either control (nontransgenic) goats' milk or milk from hLZ-transgenic goats. Growth was assessed by weight gain. Nonchallenged pigs were necropsied after 6 wk, whereas the remaining pigs were necropsied at 7 wk following bacterial challenge. We determined the numbers of total coliforms and E. coli and examined small intestinal histology for all pigs. Complete blood counts were also determined pre- and postchallenge. Challenged pigs receiving hLZ milk had fewer total coliforms (P = 0.029) and E. coli (P = 0.030) in the ileum than controls. hLZ-fed pigs also had a greater duodenal villi width (P = 0.029) than controls. Additionally, nonchallenged hLZ-fed pigs had fewer intraepithelial lymphocytes per micron of villi height (P = 0.020) than nonchallenged controls. These results indicate that the consumption of pasteurized hLZ goats' milk has the potential to improve gastrointestinal health and is protective against an EPEC in young weaned pigs. These same benefits may occur in young children if they were to consume milk from hLZ-transgenic goats. PMID- 18424603 TI - Carboxyl ester lipase from either mother's milk or the pancreas is required for efficient dietary triglyceride digestion in suckling mice. AB - Because dietary fats provide an important source of energy in the newborn, the efficient digestion of dietary fats is critical to their well-being. Despite the importance of dietary fat digestion, newborns have a deficiency of pancreatic triglyceride lipase, the predominant digestive lipase in adults. The efficient dietary fat digestion in newborns suggests that other lipases must compensate for the lack of pancreatic triglyceride lipase. In this study, we test the hypothesis that breast milk, pancreatic carboxyl ester lipase (CEL), or both contribute to dietary fat digestion in the newborn. To test this hypothesis, we determined the amount and composition of fecal fat in wild-type and CEL-deficient newborns nursed by either wild-type or CEL-deficient dams. We tested all genetic permutations of the nursing pairs. An interaction between the genotype of the dam and of the pup determined the amount of fecal fat (P < 0.001). Fecal fat was highest in CEL-deficient pups nursed by CEL-deficient dams. Furthermore, only the feces from the CEL-deficient pups nursed by CEL-deficient dams contained undigested lipids. Even with increased fecal fats, the CEL-deficient pups had normal weight gain. Our results demonstrate that CEL contributes significantly to dietary triglyceride digestion whether it originates from mother's milk or pancreatic secretions. However, only the absence of both mother's milk and pancreatic CEL produces fat maldigestion. The absence of a single CEL source makes no difference in the efficiency of dietary fat absorption. PMID- 18424604 TI - Perinatal iron deficiency affects locomotor behavior and water maze performance in adult male and female rats. AB - Iron deficiency during early growth and development adversely affects multiple facets of cognition and behavior in adult rats. The purpose of this study was to assess the nature of the learning and locomotor behavioral deficits observed in male and female rats in the absence of depressed brain iron levels at the time of testing. Adult female Wistar rats were fed either an iron-enriched diet (>225 mg/kg Fe) or an iron-restricted diet (3 mg/kg Fe) for 2 wk prior to and throughout gestation, and a nonpurified diet (270 mg/kg Fe) thereafter. Open field (OF) and Morris water maze (MWM) testing began when the offspring reached early adulthood (12 wk). At birth, perinatal iron-deficient (PID) offspring had reduced (P < 0.001) hematocrits (-33%), liver iron stores (-83%), and brain iron concentrations (-38%) compared with controls. Although there were no differences in iron status in adults, the PID males and females exhibited reduced OF exploratory behavior, albeit only PID males had an aversion to the center of the apparatus (2.5 vs. 6.9% in controls, P < 0.001). Additionally, PID males required greater path lengths to reach the hidden platform in the MWM, had reduced spatial bias for the target quadrant, and had a tendency for greater thigmotactic behavior in the probe trials (16.5 vs. 13.0% in controls; P = 0.06). PID females had slower swim speeds in all testing phases (-6.2%; P < 0.001). These results suggest that PID has detrimental programming effects in both male and female rats, although the behaviors suggest different mechanisms may be involved in each sex. PMID- 18424605 TI - Dietary phytoestrogens are not associated with risk of overall breast cancer but diets rich in coumestrol are inversely associated with risk of estrogen receptor and progesterone receptor negative breast tumors in Swedish women. AB - Results from epidemiological and experimental studies indicate that phytoestrogens may protect against breast cancer. Because one of the biological effects of phytoestrogens is probably estrogenic, it's possible that the preventive effect on breast cancer differs by estrogen receptor (ER) or progesterone receptor (PR) status of the tumor. We evaluated the associations between dietary phytoestrogen (isoflavonoids, lignans, and coumestrol) intake and risk of breast cancer and whether the ER/PR statuses of the tumor influence this relationship. In 1991-2 a prospective population-based cohort study among Swedish pre- and postmenopausal women was performed, making questionnaire data available for 45,448 women. A total of 1014 invasive breast cancers were diagnosed until December 2004. Cox proportional hazards models were performed to estimate multivariate risk ratios, 95% CI for associations with risk of breast cancer. Intakes of lignan, isoflavonoid, or coumestrol were not associated with breast cancer risk overall or before or after 50 y of age. The effects of lignans or isoflavonoids were independent of receptor status. However, intake of coumestrol was associated with decreased risk of receptor negative tumors (ER-PR-) but not positive tumors. The risk of ER-PR- tumors was significantly lower (50%) in women with intermediate coumestrol intake compared with those who did not consume any. In conclusion, we found no association between intake of isoflavonoids or lignans and breast cancer risk. Our results of a decreased risk of ER-PR- tumors in women with intermediate intake of coumestrol could be due to chance because of the low intake. The results should be confirmed in other studies. PMID- 18424606 TI - The French National Nutrition and Health Program score is associated with nutritional status and risk of major chronic diseases. AB - Few studies have found that adherence to dietary guidelines reduces the incidence of chronic disease. In 2001, a National Nutrition and Health Program (Program National Nutrition Sante) was implemented in France and included 9 quantified priority nutritional goals involving fruit, vegetable, and nutrient intakes, nutritional status, and physical activity. We developed an index score that includes indicators of these public health objectives and examined the association between this score and the incidence of major chronic diseases in the Supplementation en Vitamines et Mineraux AntioXydants cohort. Data from middle aged adults free of major chronic diseases and who provided at least 3 24-h dietary records during the first 2 y of follow-up have been included in the present analysis (n = 4,976). Major chronic disease, documented during the 8-y follow-up period (n = 455), was defined as the combination of cardiovascular disease (n = 131), cancer (n = 261), or death (n = 63), whichever came first. In fully adjusted Cox models, men in the top tertile score compared with those in the lowest one had a 36% lower risk of major chronic diseases (hazard ratio = 0.64; 95% CI: 0.44-0.96). No association was found in women. Healthy diet and lifestyle were associated with a lower risk of chronic diseases, particularly in men, thereby underlying relevance of the French nutritional recommendations. PMID- 18424607 TI - Long-chain polyunsaturated fatty acids modulate interleukin-1beta-induced changes in behavior, monoaminergic neurotransmitters, and brain inflammation in rats. AB - Recent evidence has suggested that an imbalance between membrane (n-3) and (n-6) fatty acids may contribute to the etiology of autoimmune and neurodegenerative diseases. In this study, the mechanisms by which eicosapentaenoic acid (EPA), gamma-linolenic acid (GLA), and arachidonic acid (AA) modulate neurotransmitters, behavior, and brain inflammation were evaluated in rats that received central saline or interleukin-1beta (IL-1) administrations. In rats treated with saline, only the AA-enriched diet significantly increased anxiety-like behavior in the elevated plus maze, which was associated with increased corticosterone secretion. AA also increased the turnover of dopamine (DA), noradrenaline (NA), and serotonin (5-HT) in the amygdala and increased the prostaglandin (PG)E(2) level in the hippocampus. IL-1 administration slowed rat learning in the water maze and increased anxiety-like behavior, changes which were associated with increased homovanillic acid and 5-HT turnover, decreased NA in the hippocampus and amygdala, decreased DA in the frontal cortex, and decreased IL-10 in limbic brain regions. Increased corticosterone secretion following IL-1 administration was accompanied by increased NA turnover in the hippocampus (P < 0.05) and increased PGE(2) concentration (P < 0.01) in the limbic brain regions. Of the 3 diets tested, only EPA attenuated IL-1-induced behavioral changes (P < 0.05 or 0.01), which was associated with the modulation of EPA on the neuroendocrine and immune changes induced by IL-1. GLA reduced hippocampal PGE(2) concentration in rats given IL-1 (P < 0.01). AA did not counteract any of the changes induced by IL-1. These results suggest that EPA, GLA, and AA play different roles in the neuroendocrine-immune network. PMID- 18424608 TI - Dietary energy source affects glucose kinetics in trained Arabian geldings at rest and during endurance exercise. AB - Advances in modeling and tracer techniques provide new perspective into glucose utilization and potential consequences to health or exercise performance. This study used stable isotope and compartmental modeling to evaluate how adaptation to a feed high in sugar and starch (SS) compared with a feed high in fat and fiber (FF) affects glucose kinetics at rest and during exercise in horses. Six trained Arabians adapted to each feed underwent similar tests at rest and while running approximately 4 m/s on a treadmill. For both tests, horses received 100 micromol/kg body weight [6,6-(2)H]glucose through a venous catheter. Circulating tracer glucose was described for 150 min by exponential decay curves and compartmental analysis. All parameters of glucose transfer increased with exercise (P < or = 0.004). Compared with FF horses, SS horses had higher circulating glucose (P = 0.022) and fractional glucose transfer rates (min(-1)) at rest (P = 0.055). Exercise increased glucose irreversible loss (mmol/min) more in SS horses (P = 0.037). Total glucose transfer during exercise tended to be greater in SS horses (0.027 +/- 0.002 mmol/min) compared with FF horses (0.023 +/ 0.002 mmol/min) (P = 0.109). This study characterized the effect of diet on glucose kinetics in resting and exercising horses using new modeling methods. Horses adapted to a fat-supplemented feed utilized less glucose during low intensity exercise. Fat supplementation in horses may therefore promote greater flexibility in the selection of substrate to meet energy demands for optimal health and performance. PMID- 18424609 TI - Kinetic analysis shows that vitamin A disposal rate in humans is positively correlated with vitamin A stores. AB - Vitamin A (VA) kinetics, storage, and disposal rate were determined in well nourished Chinese and U.S. adults using model-based compartmental analysis. [(2)H(8)]Retinyl acetate (8.9 micromol) was orally administered to U.S. (n = 12; 59 +/- 9 y; mean +/- SD) and Chinese adults (n = 14; 54 +/- 4 y) and serum tracer and VA concentrations were measured from 3 h to 56 d. Using the Windows version of the Simulation, Analysis and Modeling software, we determined that the average time from dosing until appearance of labeled retinol in serum was greater in U.S. subjects (40.6 +/- 8.47 h) than in Chinese subjects (32.2 +/- 5.84 h; P < 0.01). Model-predicted total traced mass (898 +/- 637 vs. 237 +/- 109 micromol), disposal rate (14.7 +/- 5.87 vs. 5.58 +/- 2.04 micromol/d), and system residence time (58.9 +/- 28.7 vs. 42.9 +/- 14.6 d) were greater in U.S. than in Chinese subjects (P < 0.05). The model-predicted VA mass and VA mass estimated by deuterated retinol dilution at 3 and 24 d did not differ. VA disposal rate was positively correlated with VA traced mass in Chinese (R(2) = 0.556), U.S. (R(2) = 0.579), and all subjects (R(2) = 0.808). Additionally, VA disposal rate was significantly correlated with serum retinol pool size (R(2) = 0.227) and retinol concentration (R(2) = 0.330) in all subjects. Our results support the hypothesis that VA stores are the principle determinant of VA disposal rate in healthy, well nourished adults. PMID- 18424610 TI - Antioxidant supplementation and risk of skin cancers. PMID- 18424611 TI - Plasma S-adenosylhomocysteine versus homocysteine as a marker for vascular disease. PMID- 18424613 TI - Distinct light-initiated gene expression and cell cycle programs in the shoot apex and cotyledons of Arabidopsis. AB - In darkness, shoot apex growth is repressed, but it becomes rapidly activated by light. We show that phytochromes and cryptochromes play largely redundant roles in this derepression in Arabidopsis thaliana. We examined the light activation of transcriptional changes in a finely resolved time course, comparing the shoot apex (meristem and leaf primordia) and the cotyledon and found >5700 differentially expressed genes. Early events specific to the shoot apices included the repression of genes for Really Interesting New Gene finger proteins and basic domain/leucine zipper and basic helix-loop-helix transcription factors. The downregulation of auxin and ethylene and the upregulation of cytokinin and gibberellin hormonal responses were also characteristic of shoot apices. In the apex, genes involved in ribosome biogenesis and protein translation were rapidly and synchronously induced, simultaneously with cell proliferation genes, preceding visible organ growth. Subsequently, the activation of signaling genes and transcriptional signatures of cell wall expansion, turgor generation, and plastid biogenesis were apparent. Furthermore, light regulates the forms and protein levels of two transcription factors with opposing functions in cell proliferation, E2FB and E2FC, through the Constitutively Photomorphogenic1 (COP1), COP9-Signalosome5, and Deetiolated1 light signaling molecules. These data provide the basis for reconstruction of the regulatory networks for light regulated meristem, leaf, and cotyledon development. PMID- 18424614 TI - Dissecting the molecular basis of the regulation of wood formation by auxin in hybrid aspen. AB - Indole acetic acid (auxin) is a key regulator of wood formation, and an observed overlap between auxin concentration gradient and developing secondary xylem cells has led to the hypothesis that auxin regulates wood formation by acting as a morphogen. We dissected the role of auxin in wood formation by identifying the auxin-responsive transcriptome in wood-forming tissues and investigating alterations in wood formation in transgenic hybrid aspen plants (Populus tremula x Populus tremuloides) with perturbed auxin signaling. We showed that auxin responsive genes in wood-forming tissues respond dynamically to changes in cellular auxin levels. However, the expression patterns of most of the auxin responsive genes displayed limited correlation with the auxin concentration across this developmental zone. Perturbing auxin signaling by reducing auxin responsiveness reduced the cambial cell division activity, caused spatial deregulation of cell division of the cambial initials, and led to reductions in not only radial but also axial dimensions of fibers and vessels. We propose that, instead of acting as a morphogen, changes in auxin concentration in developing secondary xylem cells may provide important regulatory cues that modulate the expression of a few key regulators; these, in turn, may control the global gene expression patterns that are essential for normal secondary xylem development. PMID- 18424615 TI - Arabidopsis SCARs function interchangeably to meet actin-related protein 2/3 activation thresholds during morphogenesis. AB - During polarized growth and tissue morphogenesis, cells must reorganize their cytoplasm and change shape in response to growth signals. Dynamic polymerization of actin filaments is one cellular component of polarized growth, and the actin related protein 2/3 (ARP2/3) complex is an important actin filament nucleator in plants. ARP2/3 alone is inactive, and the Arabidopsis thaliana WAVE complex translates Rho-family small GTPase signals into an ARP2/3 activation response. The SCAR subunit of the WAVE complex is the primary activator of ARP2/3, and plant and vertebrate SCARs are encoded by a small gene family. However, it is unclear if SCAR isoforms function interchangeably or if they have unique properties that customize WAVE complex functions. We used the Arabidopsis distorted group mutants and an integrated analysis of SCAR gene and protein functions to address this question directly. Genetic results indicate that each of the four SCARs functions in the context of the WAVE-ARP2/3 pathway and together they define the lone mechanism for ARP2/3 activation. Genetic interactions among the scar mutants and transgene complementation studies show that the activators function interchangeably to meet the threshold for ARP2/3 activation in the cell. Interestingly, double, triple, and quadruple mutant analyses indicate that individual SCAR genes vary in their relative importance depending on the cell type, tissue, or organ that is analyzed. Differences among SCARs in mRNA levels and the biochemical efficiency of ARP2/3 activation may explain the functional contributions of individual genes. PMID- 18424616 TI - The structure of sucrose phosphate synthase from Halothermothrix orenii reveals its mechanism of action and binding mode. AB - Sucrose phosphate synthase (SPS) catalyzes the transfer of a glycosyl group from an activated donor sugar, such as uridine diphosphate glucose (UDP-Glc), to a saccharide acceptor D-fructose 6-phosphate (F6P), resulting in the formation of UDP and D-sucrose-6'-phosphate (S6P). This is a central regulatory process in the production of sucrose in plants, cyanobacteria, and proteobacteria. Here, we report the crystal structure of SPS from the nonphotosynthetic bacterium Halothermothrix orenii and its complexes with the substrate F6P and the product S6P. SPS has two distinct Rossmann-fold domains with a large substrate binding cleft at the interdomain interface. Structures of two complexes show that both the substrate F6P and the product S6P bind to the A-domain of SPS. Based on comparative analysis of the SPS structure with other related enzymes, the donor substrate, nucleotide diphosphate glucose, binds to the B-domain of SPS. Furthermore, we propose a mechanism of catalysis by H. orenii SPS. Our findings indicate that SPS from H. orenii may represent a valid model for the catalytic domain of plant SPSs and thus may provide useful insight into the reaction mechanism of the plant enzyme. PMID- 18424617 TI - Augmented inducible nitric oxide synthase expression and increased NO production reduce sepsis-induced lung injury and mortality in myeloperoxidase-null mice. AB - The myeloperoxidase (MPO)-hydrogen peroxide-halide system is an efficient oxygen dependent antimicrobial component of polymorphonuclear leukocyte (PMN)-mediated host defense. However, MPO deficiency results in few clinical consequences indicating the activation of compensatory mechanisms. Here, we determined possible mechanisms protecting the host using MPO(-/-) mice challenged with live gram-negative bacterium Escherichia coli. We observed that MPO(-/-) mice unexpectedly had improved survival compared with wild-type (WT) mice within 5-12 h after intraperitoneal E. coli challenge. Lungs of MPO(-/-) mice also demonstrated lower bacterial colonization and markedly attenuated increases in microvascular permeability and edema formation after E. coli challenge compared with WT. However, PMN sequestration in lungs of both groups was similar. Basal inducible nitric oxide synthase (iNOS) expression was significantly elevated in lungs and PMNs of MPO(-/-) mice, and NO production was increased two- to sixfold compared with WT. Nitrotyrosine levels doubled in lungs of WT mice within 1 h after E. coli challenge but did not change in MPO(-/-) mice. Inhibition of iNOS in MPO(-/-) mice significantly increased lung edema and reduced their survival after E. coli challenge, but iNOS inhibitor had the opposite effect in WT mice. Thus augmented iNOS expression and NO production in MPO(-/-) mice compensate for the lack of HOCl-mediated bacterial killing, and the absence of MPO-derived oxidants mitigates E. coli sepsis-induced lung inflammation and injury. PMID- 18424618 TI - Does adiponectin play a role in pulmonary emphysema? PMID- 18424619 TI - Prostacyclin analogs stimulate VEGF production from human lung fibroblasts in culture. AB - Prostacyclin is a short-lived metabolite of arachidonic acid that is produced by several cells in the lung and prominently by endothelial cells. It increases intracellular cAMP levels activating downstream signaling thus regulating vascular mesenchymal cell functions. The alveolar wall contains a rich capillary network as well as a population of mesenchymal cells, i.e., fibroblasts. The current study evaluated the hypothesis that prostacyclin may mediate signaling between endothelial and mesenchymal cells in the alveolar wall by assessing the ability of prostacyclin analogs to modulate fibroblast release of VEGF. To accomplish this study, human lung fibroblasts were cultured in routine culture on plastic support and in three-dimensional collagen gels with or without three prostacyclin analogs, carbaprostacyclin, iloprost, and beraprost, and the production of VEGF was evaluated by ELISA and quantitative real-time PCR. Iloprost and beraprost significantly stimulated VEGF mRNA levels and protein release in a concentration-dependent manner. These effects were blocked by the adenylate cyclase inhibitor SQ-22536 and by the protein kinase A (PKA) inhibitor KT-5720 and were reproduced by a direct PKA activator but not by an activator of exchange protein directly activated by cAMP (Epac), indicating that cAMP activated PKA signaling mediated the effect. Since VEGF serves to maintain the pulmonary microvasculature, the current study suggests that prostacyclin is part of a bidirectional signaling network between the mesenchymal and vascular cells of the alveolar wall. Prostacyclin analogs, therefore, have the potential to modulate the maintenance of the pulmonary microcirculation by driving the production of VEGF from lung fibroblasts. PMID- 18424620 TI - Overexpression of the Na-K-ATPase alpha2-subunit improves lung liquid clearance during ventilation-induced lung injury. AB - Mechanical ventilation with high tidal volumes (HV(T)) impairs lung liquid clearance (LLC) and downregulates alveolar epithelial Na-K-ATPase. We have previously reported that the Na-K-ATPase alpha(2)-subunit contributes to LLC in normal rat lungs. Here we tested whether overexpression of Na-K-ATPase alpha(2) subunit in the alveolar epithelium would increase clearance in a HV(T) model of lung injury. We infected rat lungs with a replication-incompetent adenovirus that expresses Na-K-ATPase alpha(2)-subunit gene (Adalpha(2)) 7 days before HV(T) mechanical ventilation. HV(T) ventilation decreased LLC by approximately 50% in untreated, sham, and Adnull-infected rats. Overexpression of Na-K-ATPase alpha(2) subunit prevented the decrease in clearance caused by HV(T) and was associated with significant increases in Na-K-ATPase alpha(2) protein abundance and activity in peripheral lung basolateral membrane fractions. Ouabain at 10(-5) M, a concentration that inhibits the alpha(2) but not the Na-K-ATPase alpha(1), decreased LLC in Adalpha(2)-infected rats to the same level as sham and Adnull infected lungs, suggesting that the increased clearance in Adalpha(2) lungs was due to Na-K-ATPase alpha(2) expression and activity. In summary, we provide evidence that augmentation of the Na-K-ATPase alpha(2)-subunit, via gene transfer, may accelerate LLC in the injured lung. PMID- 18424621 TI - Differences in STIM1 and TRPC expression in proximal and distal pulmonary arterial smooth muscle are associated with differences in Ca2+ responses to hypoxia. AB - Hypoxic pulmonary vasoconstriction (HPV) requires Ca(2+) influx through store operated Ca(2+) channels (SOCC) in pulmonary arterial smooth muscle cells (PASMC) and is greater in distal than proximal pulmonary arteries (PA). SOCC may be composed of canonical transient receptor potential (TRPC) proteins and activated by stromal interacting molecule 1 (STIM1). To assess the possibility that HPV is greater in distal PA because store-operated Ca(2+) entry (SOCE) is greater in distal PASMC, we measured intracellular Ca(2+) concentration ([Ca(2+)](i)) and SOCE in primary cultures of PASMC using fluorescent microscopy and the Ca(2+) sensitive dye fura 2. Both hypoxia (4% O(2)) and KCl (60 mM) increased [Ca(2+)](i). Responses to hypoxia, but not KCl, were greater in distal cells. We measured SOCE in PASMC perfused with Ca(2+)-free solutions containing cyclopiazonic acid to deplete Ca(2+) stores in sarcoplasmic reticulum and nifedipine to prevent Ca(2+) entry through L-type voltage-operated Ca(2+) channels. Under these conditions, the increase in [Ca(2+)](i) caused by restoration of extracellular Ca(2+) and the decrease in fura 2 fluorescence caused by Mn(2+) were greater in distal PASMC, indicating greater SOCE. Moreover, the increase in SOCE caused by hypoxia was also greater in distal cells. Real time quantitative polymerase chain reaction analysis of PASMC and freshly isolated deendothelialized PA tissue demonstrated expression of STIM1 and five of seven known TRPC isoforms (TRPC1 > TRPC6 > TRPC4 >> TRPC3 approximately TRPC5). For both protein, as measured by Western blotting, and mRNA, expression of STIM1, TRPC1, TRPC6, and TRPC4 was greater in distal than proximal PASMC and PA. These results provide further support for the importance of SOCE in HPV and suggest that HPV is greater in distal than proximal PA because greater numbers and activation of SOCC in distal PASMC generate bigger increases in [Ca(2+)](i). PMID- 18424622 TI - Proteasome inhibition improves diaphragm function in congestive heart failure rats. AB - In congestive heart failure (CHF), diaphragm weakness is known to occur and is associated with myosin loss and activation of the ubiquitin-proteasome pathway. The effect of modulating proteasome activity on myosin loss and diaphragm function is unknown. The present study investigated the effect of in vivo proteasome inhibition on myosin loss and diaphragm function in CHF rats. Coronary artery ligation was used as an animal model for CHF. Sham-operated rats served as controls. Animals were treated with the proteasome inhibitor bortezomib (intravenously) or received saline (0.9%) injections. Force generating capacity, cross-bridge cycling kinetics, and myosin content were measured in diaphragm single fibers. Proteasome activity, caspase-3 activity, and MuRF-1 and MAFbx mRNA levels were determined in diaphragm homogenates. Proteasome activities in the diaphragm were significantly reduced by bortezomib. Bortezomib treatment significantly improved diaphragm single fiber force generating capacity (approximately 30-40%) and cross-bridge cycling kinetics (approximately 20%) in CHF. Myosin content was approximately 30% higher in diaphragm fibers from bortezomib-treated CHF rats than saline. Caspase-3 activity was decreased in diaphragm homogenates from bortezomib-treated rats. CHF increased MuRF-1 and MAFbx mRNA expression in the diaphragm, and bortezomib treatment diminished this rise. The present study demonstrates that treatment with a clinically used proteasome inhibitor improves diaphragm function by restoring myosin content in CHF. PMID- 18424623 TI - EGF receptor tyrosine kinase inhibitors diminish transforming growth factor-alpha induced pulmonary fibrosis. AB - Transforming growth factor-alpha (TGF-alpha) is a ligand for the EGF receptor (EGFR). EGFR activation is associated with fibroproliferative processes in human lung disease and animal models of pulmonary fibrosis. We determined the effects of EGFR tyrosine kinase inhibitors gefitinib (Iressa) and erlotinib (Tarceva) on the development and progression of TGF-alpha-induced pulmonary fibrosis. Using a doxycycline-regulatable transgenic mouse model of lung-specific TGF-alpha expression, we determined effects of treatment with gefitinib and erlotinib on changes in lung histology, total lung collagen, pulmonary mechanics, pulmonary hypertension, and expression of genes associated with synthesis of ECM and vascular remodeling. Induction in the lung of TGF-alpha caused progressive pulmonary fibrosis over an 8-wk period. Daily administration of gefitinib or erlotinib prevented development of fibrosis, reduced accumulation of total lung collagen, prevented weight loss, and prevented changes in pulmonary mechanics. Treatment of mice with gefitinib 4 wk after the induction of TGF-alpha prevented further increases in and partially reversed total collagen levels and changes in pulmonary mechanics and pulmonary hypertension. Increases in expression of genes associated with synthesis of ECM as well as decreases of genes associated with vascular remodeling were also prevented or partially reversed. Administration of gefitinib or erlotinib did not cause interstitial fibrosis or increases in lavage cell counts. Administration of small molecule EGFR tyrosine kinase inhibitors prevented further increases in and partially reversed pulmonary fibrosis induced directly by EGFR activation without inducing inflammatory cell influx or additional lung injury. PMID- 18424625 TI - Evidence for the role of metabotropic glutamate (mGlu)2 not mGlu3 receptors in the preclinical antipsychotic pharmacology of the mGlu2/3 receptor agonist (-) (1R,4S,5S,6S)-4-amino-2-sulfonylbicyclo[3.1.0]hexane-4,6-dicarboxylic acid (LY404039). AB - (-)-(1R,4S,5S,6S)-4-amino-2-sulfonylbicyclo[3.1.0]hexane-4,6-dicarboxylic acid (LY404039) is a potent and selective group II metabotropic glutamate [(mGlu)2 and mGlu3] receptor agonist for which its prodrug LY2140023 [(1R,4S,5S,6S)-2 thiabicyclo[3.1.0]-hexane-4,6-dicarboxylic acid,4-[(2S)-2-amino-4-(methylthio)-1 oxobutyl]amino-, 2,2-dioxide monohydrate] has recently been shown to have efficacy in the treatment of the positive and negative symptoms of schizophrenia. In this article, we use mGlu receptor-deficient mice to investigate the relative contribution of mGlu2 and mGlu3 receptors in mediating the antipsychotic profile of LY404039 in the phencyclidine (PCP) and d-amphetamine (AMP) models of psychosis. To further explore the mechanism of action of LY404039, we compared the drugs' ability to block PCP-induced hyperlocomotion to that of atypical antipsychotics in wild-type and mice lacking mGlu2/3 receptors. In wild-type animals, LY404039 (3-30 mg/kg i.p.) significantly reversed AMP (5 mg/kg, i.p.) induced increases in ambulations, distance traveled, and reduced time spent at rest. LY404039 reversed PCP (7.5 mg/kg i.p.)-evoked behaviors at 10 mg/kg. The antipsychotic-like effects of LY404039 (10 mg/kg i.p.) on PCP and AMP-evoked behavioral activation were absent in mGlu2 and mGlu2/3 but not in mGlu3 receptor deficient mice, indicating that the activation of mGlu2 and not mGlu3 receptors is responsible for the antipsychotic-like effects of the mGlu2/3 receptor agonist LY404039. In contrast, the atypical antipsychotic drugs clozapine and risperidone inhibited PCP-evoked behaviors in both wild-type and mGlu2/3 receptor-deficient mice. These data demonstrate that the antipsychotic-like effects of the mGlu2/3 receptor agonist LY404039 in psychostimulant models of psychosis are mechanistically distinct from those of atypical antipsychotic drugs and are dependent on functional mGlu2 and not mGlu3 receptors. PMID- 18424626 TI - The absence of ALTERNATIVE OXIDASE1a in Arabidopsis results in acute sensitivity to combined light and drought stress. AB - Treatment of Arabidopsis (Arabidopsis thaliana) alternative oxidase1a (aox1a) mutant plants with moderate light under drought conditions resulted in a phenotypic difference compared with ecotype Columbia (Col-0), as evidenced by a 10-fold increase in the accumulation of anthocyanins in leaves, alterations in photosynthetic efficiency, and increased superoxide radical and reduced root growth at the early stages of seedling growth. Analysis of metabolite profiles revealed significant changes upon treatment in aox1a plants typical of combined stress treatments, and these were less pronounced or absent in Col-0 plants. These changes were accompanied by alteration in the abundance of a variety of transcripts during the stress treatment, providing a molecular fingerprint for the stress-induced phenotype of aox1a plants. Transcripts encoding proteins involved in the synthesis of anthocyanins, transcription factors, chloroplastic and mitochondrial components, cell wall synthesis, and sucrose and starch metabolism changed, indicating that effects were not confined to mitochondria, where the AOX1a protein is located. Microarray and quantitative reverse transcription-polymerase chain reaction analysis revealed that transcripts typically induced upon stress treatment or involved in antioxidant defense systems, especially chloroplast-located antioxidant defense components, had altered basal levels in untreated aox1a plants, suggesting a significant change in the basal equilibrium of signaling pathways that regulate these components. Taken together, these results indicate that aox1a plants have a greatly altered stress response even when mitochondria or the mitochondrial electron transport chain are not the primary target of the stress and that AOX1a plays a broad role in determining the normal redox balance in the cell. PMID- 18424627 TI - Transcript profiling reveals new insights into the acclimation of the mesophilic fresh-water cyanobacterium Synechococcus elongatus PCC 7942 to iron starvation. AB - The regulatory network for acclimation of the obligate photoautotrophic fresh water cyanobacterium Synechococcus elongatus PCC 7942 to iron (Fe) limitation was studied by transcript profiling with an oligonucleotide whole genome DNA microarray. Six regions on the chromosome with several Fe-regulated genes each were identified. The irpAB and fut region encode putative Fe uptake systems, the suf region participates in [Fe-sulfur] cluster assembly under oxidative stress and Fe limitation, the isiAB region encodes CP43' and flavodoxin, the idiCB region encodes the NuoE-like electron transport associated protein IdiC and the transcriptional activator IdiB, and the ackA/pgam region encodes an acetate kinase and a phosphoglycerate mutase. We also investigated the response of two S. elongatus PCC 7942 mutants to Fe starvation. These were mutant K10, lacking IdiB but containing IdiC, and mutant MuD, representing a idiC-merodiploid mutant with a strongly reduced amount of IdiC as well as IdiB. The absence of IdiB in mutant K10 or the strongly reduced amount of IdiB in mutant MuD allowed for the identification of additional members of the Fe-responsive IdiB regulon. Besides idiA and the irpAB operon somB(1), somA(2), ftr1, ackA, pgam, and nat also seem to be regulated by IdiB. In addition to the reduced amount of IdiB in MuD, the low concentration of IdiC may be responsible for a number of additional changes in the abundance of mainly photosynthesis-related transcripts as compared to the wild type and mutant K10. This fact may explain why it has been impossible to obtain a fully segregated IdiC-free mutant, whereas it was possible to obtain a fully segregated IdiB-free mutant. PMID- 18424628 TI - Effect of chronic and selective endothelin receptor antagonism on microvascular function in type 2 diabetes. AB - Vascular dysfunction, which presents either as an increased response to vasoconstrictors or an impaired relaxation to dilator agents, results in worsened cardiovascular outcomes in diabetes. We have established that the mesenteric circulation in Type 2 diabetes is hyperreactive to the potent vasoconstrictor endothelin-1 (ET-1) and displays increased nitric oxide-dependent vasodilation. The current study examined the individual and/or the relative roles of the ET receptors governing vascular function in the Goto-Kakizaki rat, a mildly hyperglycemic, normotensive, and nonobese model of Type 2 diabetes. Diabetic and control rats received an antagonist to either the ET type A (ETA; atrasentan; 5 mg x kg(-1) x day(-1)) or type B (ET(B); A-192621; 15 or 30 mg x kg(-1) x day( 1)) receptors for 4 wk. Third-order mesenteric arteries were isolated, and vascular function was assessed with a wire myograph. Maximum response to ET-1 was increased in diabetes and attenuated by ETA antagonism. ETB blockade with 15 mg/kg A-192621 augmented vasoconstriction in controls, whereas it had no further effect on ET-1 hyperreactivity in diabetes. The higher dose of A-192621 showed an ETA-like effect and decreased vasoconstriction in diabetes. Maximum relaxation to acetylcholine (ACh) was similar across groups and treatments. ETB antagonism at either dose had no effect on vasorelaxation in control rats, whereas in diabetes the dose-response curve to ACh was shifted to the right, indicating a decreased relaxation at 15 mg/kg A-192621. These results suggest that ETA receptor blockade attenuates vascular dysfunction and that ETB receptor antagonism exhibits differential effects depending on the dose of the antagonists and the disease state. PMID- 18424629 TI - Transmural distribution of metabolic abnormalities and glycolytic activity during dobutamine-induced demand ischemia. AB - The heterogeneity across the left ventricular wall is characterized by higher rates of oxygen consumption, systolic thickening fraction, myocardial perfusion, and lower energetic state in the subendocardial layers (ENDO). During dobutamine stimulation-induced demand ischemia, the transmural distribution of energy demand and metabolic markers of ischemia are not known. In this study, hemodynamics, transmural high-energy phosphate (HEP), 2-deoxyglucose-6-phosphate (2-DGP) levels, and myocardial blood flow (MBF) were determined under basal conditions, during dobutamine infusion (DOB: 20 microg x kg(-1) x min(-1) iv), and during coronary stenosis + DOB + 2-deoxyglucose (2-DG) infusion. DOB increased rate pressure products (RPP) and MBF significantly without affecting the subendocardial-to-subepicardial blood flow ratio (ENDO/EPI) or HEP levels. During coronary stenosis + DOB + 2-DG infusion, RPP, ischemic zone (IZ) MBF, and ENDO/EPI decreased significantly. The IZ ratio of creatine phosphate-to-ATP decreased significantly [2.30 +/- 0.14, 2.06 +/- 0.13, and 2.04 +/- 0.11 to 1.77 +/- 0.12, 1.70 +/- 0.11, and 1.72 +/- 0.12 for EPI, midmyocardial (MID), and ENDO, respectively], and 2-DGP accumulated in all layers, as evidenced by the 2 DGP/PCr (0.55 +/- 0.12, 0.52 +/- 0.10, and 0.37 +/- 0.08 for EPI, MID, and ENDO, respectively; P < 0.05, EPI > ENDO). In the IZ the wet weight-to-dry weight ratio was significantly increased compared with the normal zone (5.9 +/- 0.5 vs. 4.4 +/ 0.4; P < 0.05). Thus, in the stenotic perfused bed, during dobutamine-induced high cardiac work state, despite higher blood flow, the subepicardial layers showed the greater metabolic changes characterized by a shift toward higher carbohydrate metabolism, suggesting that a homeostatic response to high-cardiac work state is characterized by more glucose utilization in energy metabolism. PMID- 18424630 TI - Coronary nitric oxide production controls cardiac substrate metabolism during pregnancy in the dog. AB - The aim of this study was to examine the role of nitric oxide (NO) in the control of cardiac metabolism at 60 days of pregnancy (P60) in the dog. There was a basal increase in diastolic coronary blood flow during pregnancy and a statistically significant increase in cardiac output (55 +/- 4%) and in cardiac NOx production (44 +/- 4 to 59 +/- 3 nmol/min, P < 0.05). Immunohistochemistry of the left ventricle showed an increase in endothelial nitric oxide synthase staining in the endothelial cells at P60. NO-dependent coronary vasodilation (Bezold-Jarisch reflex) was increased by 20% and blocked by N(G)-nitro-l-arginine methyl ester (l NAME). Isotopically labeled substrates were infused to measure oleate, glucose uptake, and oxidation. Glucose oxidation was not significantly different in P60 hearts (5.4 +/- 0.5 vs. 6.2 +/- 0.4 micromol/min) but greatly increased in response to l-NAME injection (to 19.9 +/- 0.9 micromol/min, P < 0.05). Free fatty acid (FFA) oxidation was increased in P60 (from 5.3 +/- 0.6 to 10.4 +/- 0.5 micromol/min, P < 0.05) and decreased in response to l-NAME (to 4.5 +/- 0.5 micromol/min, P < 0.05). There was an increased oxidation of FFA for ATP production but no change in the respiratory quotient during pregnancy. Genes associated with glucose and glycogen metabolism were downregulated, whereas genes involved in FFA oxidation were elevated. The acute inhibition of NO shifts the heart away from FFA and toward glucose metabolism despite the downregulation of the carbohydrate oxidative pathway. The increase in endothelium-derived NO during pregnancy results in a tonic inhibition of glucose oxidation and reliance on FFA uptake and oxidation to support ATP synthesis in conjunction with upregulation of FFA metabolic enzymes. PMID- 18424631 TI - Exogenous stromal cell-derived factor-1 induces modest leukocyte recruitment in vivo. AB - Stromal cell-derived factor-1 (SDF-1; CXCL12), a CXC chemokine, has been found to be involved in inflammation models in vivo and in cell adhesion, migration, and chemotaxis in vitro. This study aimed to determine whether exogenous SDF-1 induces leukocyte recruitment in mice. After systemic administration of SDF 1alpha, expression of the adhesion molecules P-selectin and VCAM-1 in mice was measured using a quantitative dual-radiolabeled Ab assay and leukocyte recruitment in various tissues was evaluated using intravital microscopy. The effect of local SDF-1alpha on leukocyte recruitment was also determined in cremaster muscle and compared with the effect of the cytokine TNFalpha and the CXC chemokine keratinocyte-derived chemokine (KC; CXCL1). Systemic administration of SDF-1alpha (10 microg, 4-5 h) induced upregulation of P-selectin, but not VCAM 1, in most tissues in mice. It caused modest leukocyte recruitment responses in microvasculature of cremaster muscle, intestine, and brain, i.e., an increase in flux of rolling leukocytes in cremaster muscle and intestines, leukocyte adhesion in all three tissues, and emigration in cremaster muscle. Local treatment with SDF-1alpha (1 microg, 4-5 h) reduced leukocyte rolling velocity and increased leukocyte adhesion and emigration in cremasteric venules, but the responses were much less profound than those elicited by KC or TNFalpha. SDF-1alpha-induced recruitment was dependent on endothelial P-selectin, but not P-selectin on platelets. We conclude that the exogenous SDF-1alpha enhances leukocyte endothelial cell interactions and induces modest and endothelial P-selectin dependent leukocyte recruitment. PMID- 18424632 TI - Oxidative stress contributes to pulmonary hypertension in the transgenic (mRen2)27 rat. AB - The transgenic (mRen2)27 (Ren2) rat overexpresses mouse renin in extrarenal tissues, causing increased local synthesis of ANG II, oxidative stress, and hypertension. However, little is known about the role of oxidative stress induced by the tissue renin-angiotensin system (RAS) as a contributing factor in pulmonary hypertension (PH). Using male Ren2 rats, we test the hypothesis that lung tissue RAS overexpression and resultant oxidative stress contribute to PH and pulmonary vascular remodeling. Mean arterial pressure (MAP), right ventricular systolic pressure (RVSP), and wall thickness of small pulmonary arteries (PA), as well as intrapulmonary NADPH oxidase activity and subunit protein expression and reactive oxygen species (ROS), were compared in age matched Ren2 and Sprague-Dawley (SD) rats pretreated with the SOD/catalase mimetic tempol for 21 days. In placebo-treated Ren2 rats, MAP and RVSP, as well as intrapulmonary NADPH oxidase activity and subunits (Nox2, p22phox, and Rac-1) and ROS, were elevated compared with placebo-treated SD rats (P < 0.05). Tempol decreased RVSP (P < 0.05), but not MAP, in Ren2 rats. Tempol also reduced intrapulmonary NADPH oxidase activity, Nox2, p22phox, and Rac-1 protein expression, and ROS in Ren2 rats (P < 0.05). Compared with SD rats, the cross sectional surface area of small PA was 38% greater (P < 0.001) and luminal surface area was 54% less (P < 0.001) in Ren2 rats. Wall surface area was reduced and luminal area was increased in tempol-treated SD and Ren2 rats compared with untreated controls (P < 0.05). Collectively, the results of this investigation support a seminal role for enhanced tissue RAS/oxidative stress as factors in development of PH and pulmonary vascular remodeling. PMID- 18424633 TI - Structure and torsion in the normal and situs inversus totalis cardiac left ventricle. II. Modeling cardiac adaptation to mechanical load. AB - Mathematical models provide a suitable platform to test hypotheses on the relation between local mechanical stimuli and responses to cardiac structure and geometry. In the present model study, we tested hypothesized mechanical stimuli and responses in cardiac adaptation to mechanical load on their ability to estimate a realistic myocardial structure of the normal and situs inversus totalis (SIT) left ventricle (LV). In a cylindrical model of the LV, 1) mass was adapted in response to myofiber strain at the beginning of ejection and to global contractility (average systolic pressure), 2) cavity volume was adapted in response to fiber strain during ejection, and 3) myofiber orientations were adapted in response to myofiber strain during ejection and local misalignment between neighboring tissue parts. The model was able to generate a realistic normal LV geometry and structure. In addition, the model was also able to simulate the instigating situation in the rare SIT LV with opposite torsion and transmural courses in myofiber direction between the apex and base [Delhaas et al. (6)]. These results substantiate the importance of mechanical load in the formation and maintenance of cardiac structure and geometry. Furthermore, in the model, adapted myocardial architecture was found to be insensitive to fiber misalignment in the transmural direction, i.e., myofiber strain during ejection was sufficient to generate a realistic transmural variation in myofiber orientation. In addition, the model estimates that, despite differences in structure, global pump work and the mass of the normal and SIT LV are similar. PMID- 18424634 TI - Effective arterial elastance as an index of pulmonary vascular load. AB - The aim of this study was to test whether the simple ratio of right ventricular (RV) end-systolic pressure (Pes) to stroke volume (SV), known as the effective arterial elastance (Ea), provides a valid assessment of pulmonary arterial load in case of pulmonary embolism- or endotoxin-induced pulmonary hypertension. Ventricular pressure-volume (PV) data (obtained with conductance catheters) and invasive pulmonary arterial pressure and flow waveforms were simultaneously recorded in two groups of six pure Pietran pigs, submitted either to pulmonary embolism (group A) or endotoxic shock (group B). Measurements were obtained at baseline and each 30 min after injection of autologous blood clots (0.3 g/kg) in the superior vena cava in group A and after endotoxin infusion in group B. Two methods of calculation of pulmonary arterial load were compared. On one hand, Ea provided by using three-element windkessel model (WK) of the pulmonary arterial system [Ea(WK)] was referred to as standard computation. On the other hand, similar to the systemic circulation, Ea was assessed as the ratio of RV Pes to SV [Ea(PV) = Pes/SV]. In both groups, although the correlation between Ea(PV) and Ea(WK) was excellent over a broad range of altered conditions, Ea(PV) systematically overestimated Ea(WK). This offset disappeared when left atrial pressure (Pla) was incorporated into Ea [Ea * (PV) = (Pes - Pla)/SV]. Thus Ea * (PV), defined as the ratio of RV Pes minus Pla to SV, provides a convenient, useful, and simple method to assess the pulmonary arterial load and its impact on the RV function. PMID- 18424636 TI - Inosine and equilibrative nucleoside transporter 2 contribute to hypoxic preconditioning in the murine cardiomyocyte HL-1 cell line. AB - The purine nucleoside adenosine is a physiologically important molecule in the heart. Brief exposure of cardiomyocytes to hypoxic challenge results in the production of extracellular adenosine, which then interacts with adenosine receptors to activate compensatory signaling pathways that lead to cellular resistance to subsequence hypoxic challenge. This phenomenon is known as preconditioning (PC), and, while adenosine is clearly involved, other components of the response are less well understood. Flux of nucleosides, such as adenosine and inosine, across cardiomyocyte membranes is dependent on equilibrative nucleoside transporters 1 and 2 (ENT1 and ENT2). We have previously shown in the murine cardiomyocyte HL-1 cell line that hypoxic challenge leads to an increase in intracellular adenosine, which exits the cell via ENT1 and preconditions via A1 and A3 adenosine receptor-dependent mechanisms. However, the role and contribution of inosine and ENT2 are unclear. In this study, we confirmed that ENT1 and ENT2 are both capable of transporting inosine. Moreover, we found that hypoxic challenge leads to a significant increase in levels of intracellular inosine, which exits the cell via both ENT1 and ENT2. Exogenously added inosine (5 microM) preconditions cardiomyocytes in an A1 adenosine receptor-dependent manner since preconditioning can be blocked by the A1 adenosine receptor antagonist 8-cyclopentyl-1,3-dipropylxanthine (1 microM) but not the A3 adenosine receptor antagonist MRS-1220 (200 nM). These data suggest that cardiomyocyte responses to hypoxic PC are more complex than previously thought, involving both adenosine and inosine and differing, but overlapping, contributions of the two ENT isoforms. PMID- 18424635 TI - Angiotensin II increases GABAB receptor expression in nucleus tractus solitarii of rats. AB - Increasing evidence indicates that both the angiotensin II (ANG II) and gamma aminobutyric acid (GABA) systems play a very important role in the regulation of blood pressure (BP). However, there is little information concerning the interactions between these two systems in the nucleus tractus solitarii (NTS). In the present study, we examined the effects of ANG II on GABAA and GABAB receptor (GAR and GBR) expression in the NTS of Sprague-Dawley rats. The direct effect of ANG II on GBR expression was determined in neurons cultured from NTS. Treatment of neuronal cultures with ANG II (100 nM, 5 h) induced a twofold increase in GBR1 expression, as detected with real-time RT-PCR and Western blots, but had no effect on GBR2 or GAR expression. In electrophysiological experiments, perfusion of neuronal cultures with the GBR agonist baclofen decreased neuronal firing rate by 39% and 63% in neurons treated with either PBS (control) or ANG II, respectively, indicating that chronic ANG II treatment significantly enhanced the neuronal response to GBR activation. In contrast, ANG II had no significant effect on the inhibitory action of the GAR agonist muscimol. In whole animal studies, intracerebroventricular infusion of ANG II induced a sustained increase in mean BP and an elevation of GBR1 mRNA and protein levels in the NTS. These results indicate that ANG II stimulates GBR expression in NTS neurons, and this could contribute to the central nervous system actions of ANG II that result in dampening of baroreflexes and elevated BP in the central actions of ANG II. PMID- 18424638 TI - The diastatic pressure-volume relationship is not the same as the end-diastolic pressure-volume relationship. AB - The end-diastolic pressure-volume (P-V) relationship (EDPVR) is routinely used to determine the passive left ventricular (LV) stiffness, although the diastatic P-V relationship (D-PVR) has also been measured. Based on the physiological difference between diastasis (the LV and atrium are relaxed and static) and end diastole (LV volume increased by atrial systole and the atrium is contracted), we hypothesized that, although both D-PVR and EDPVR include LV chamber stiffness information, they are two different, distinguishable P-V relations. Cardiac catheterization determined LV pressures, and conductance volumes in 31 subjects were analyzed. Physiological, beat-to-beat variation of the diastatic and end diastolic P-V points were fit by linear and exponential functions to generate the D-PVR and EDPVR. The extrapolated exponential D-PVR underestimated LVEDP in 82% of the heart beats (P < 0.001). The extrapolated EDPVR overestimated pressure at diastasis in 84% of the heart beats (P < 0.001). If each subject's diastatic and end-diastolic P-V data were combined to form a continuous data set to be fit by one exponential relation, the goodness of fit was always worse than if the diastatic and end-diastolic data were grouped separately and fit by two distinct exponential relations. Diastatic chamber stiffness was less than EDPVR stiffness (defined by the slope of P-V relation) for all 31 subjects (0.16 +/- 0.11 vs. 0.24 +/- 0.15 mmHg/ml, P < 0.001). We conclude that the D-PVR and EDPVR are distinguishable. Because it is not coupled to a contracted atrium, the D-PVR conveys passive LV stiffness better than the EDPVR. Additional studies that fully elucidate the physiology and biology of diastasis in health and disease are in progress. PMID- 18424637 TI - Vasoprotective effects of resveratrol and SIRT1: attenuation of cigarette smoke induced oxidative stress and proinflammatory phenotypic alterations. AB - The dietary polyphenolic compound resveratrol, by activating the protein deacetylase enzyme silent information regulator 2/sirtuin 1 (SIRT1), prolongs life span in evolutionarily distant organisms and may mimic the cytoprotective effects of dietary restriction. The present study was designed to elucidate the effects of resveratrol on cigarette smoke-induced vascular oxidative stress and inflammation, which is a clinically highly relevant model of accelerated vascular aging. Cigarette smoke exposure of rats impaired the acetylcholine-induced relaxation of carotid arteries, which could be prevented by resveratrol treatment. Smoking and in vitro treatment with cigarette smoke extract (CSE) increased reactive oxygen species production in rat arteries and cultured coronary arterial endothelial cells (CAECs), respectively, which was attenuated by resveratrol treatment. The smoking-induced upregulation of inflammatory markers (ICAM-1, inducible nitric oxide synthase, IL-6, and TNF-alpha) in rat arteries was also abrogated by resveratrol treatment. Resveratrol also inhibited CSE-induced NF-kappaB activation and inflammatory gene expression in CAECs. In CAECs, the aforementioned protective effects of resveratrol were abolished by knockdown of SIRT1, whereas the overexpression of SIRT1 mimicked the effects of resveratrol. Resveratrol treatment of rats protected aortic endothelial cells against cigarette smoking-induced apoptotic cell death. Resveratrol also exerted antiapoptotic effects in CSE-treated CAECs, which could be abrogated by knockdown of SIRT1. Resveratrol treatment also attenuated CSE-induced DNA damage in CAECs (comet assay). Thus resveratrol and SIRT1 exert antioxidant, anti-inflammatory, and antiapoptotic effects, which protect the endothelial cells against the adverse effects of cigarette smoking-induced oxidative stress. The vasoprotective effects of resveratrol will likely contribute to its antiaging action in mammals and may be especially beneficial in pathophysiological conditions associated with accelerated vascular aging. PMID- 18424639 TI - Anoxic cell core can promote necrotic cell death in cardiomyocytes at physiological extracellular PO2. AB - The physical law of diffusion imposes O2 concentration gradients from the plasma membrane to the center of the cell. The present study was undertaken to determine how such intracellular radial gradients of O2 affect the fate of isolated single cardiomyocytes. In single rat cardiomyocytes, mitochondrial respiration was moderately elevated by an oxidative phosphorylation uncoupler to augment the intracellular O2 gradient. At physiological extracellular O2 levels (2-5%), decreases in myoglobin O2 saturation and increases in NADH fluorescence at the center of the cell were imaged (anoxic cell core) while the mitochondrial membrane potential (DeltaPsim) and ATP levels at the anoxic cell core were relatively sustained. In contrast, treatment with 0.5 mM iodoacetamide (IA) to inhibit creatine kinase (CK) resulted in depletion of both DeltaPsim and ATP at the anoxic cell core. Even at normal extracellular Po2, actively respiring cardiomyocytes developed rigor contracture followed by necrotic cell death. Furthermore, such rigor was remarkably accelerated by IA, whereas cell injury was perfectly rescued by mitochondrial F1Fo inhibition by oligomycin. These results suggest that increases in radial gradients of O2 potentially promote cell death through the reverse action of F1Fo in mitochondria located at the anoxic cell core. However, in the intact cardiomyocyte, the CK-mediated energy flux from the subsarcolemmal space may sustain DeltaPsim at the cell core, thus avoiding uncontrolled consumption of ATP that can lead to necrotic cell death. Mitochondria at the anoxic core can cause necrotic cell death in cardiomyocytes at physiological extracellular Po2. PMID- 18424640 TI - A novel mouse model of X-linked cardiac hypertrophy. AB - We recovered a novel mouse mutant exhibiting neonatal lethality associated with severe fetal cardiac hypertrophy and with some adult mice dying suddenly with left ventricular hypertrophic cardiomyopathy. Using Doppler echocardiography, we screened surviving adult mice in this mutant line for cardiac hypertrophy. Cardiac dimensions were obtained either from two-dimensional images collected using a novel ECG-gated ultra-high-frequency ultrasound system or by traditional M-mode imaging on a clinical ultrasound system. These analyses identified, among the littermates, two populations of mice: those with apparent cardiac hypertrophy with hypercontractile function characterized by ejection fraction of 75-80%, and normal littermates with ejection fraction of 53-55%. Analysis of the ECG-gated two-dimensional cines indicated that the hypertrophy was of the nonobstructive type. Further analysis of heart-to-body weight ratio confirmed the ultrasound diagnosis of left ventricular hypertrophic cardiomyopathy. Histopathology showed increased ventricular wall thickness, enlarged myocyte size, and mild myofiber disarray. Ultrastructural analysis by electron microscopy revealed mitochondria hyperproliferation and dilated sarcoplasmic reticulum. Genome scanning using microsatellite DNA markers mapped the mutation to the X chromosome. DNA sequencing showed no mutations in the coding regions of several candidate genes on the X chromosome, including several known to be associated with left ventricular hypertrophic cardiomyopathy. These findings suggest that this mouse line may harbor a mutation in a novel gene causing X-linked cardiomyopathy. PMID- 18424641 TI - Apelin decreases the SR Ca2+ content but enhances the amplitude of [Ca2+]i transient and contractions during twitches in isolated rat cardiac myocytes. AB - Apelin has been reported to have a positive inotropic action in the isolated rat heart. However, the effect of apelin on sarcoplasmic reticulum (SR) Ca2+ content and its influence on intracellular Ca2+ transient during excitation-contraction coupling remains poorly understood. In the present study, we determined the effect of apelin on Ca2+ transient and contractions in isolated rat cardiomyocytes. When compared with control, treatment with apelin caused a 55.7 +/- 13.9% increase in sarcomere fraction shortening and a 43.6 +/- 4.56% increase in amplitude of electrical-stimulated intracellular Ca2+ concentration (E[Ca2+]i) transients (n = 14, P < 0.05). But SR Ca2+ content measured by caffeine-induced [Ca2+]i (C[Ca2+]i) transient was decreased 8.41 +/- 0.92% in response to apelin (n = 14, P < 0.05). Na+/Ca2+ exchanger (NCX) function was increased since half decay time of C[Ca2+]i was decreased 16.22 +/- 1.36% in response to apelin. Sarco(endo)plasmic reticulum Ca2+-ATPase (SERCA) activity was also increased by apelin. These responses can be partially or completely blocked by chelerythrine chloride, a PKC inhibitor. In addition, to confirm our data, we used indo-1 as another Ca2+ indicator and rapid cooling as another way to measure SR Ca2+ content, and we observed similar results. So we conclude that apelin has a positive inotropic effect on isolated myocytes, and increased amplitude of E[Ca2+]i is at least partially involved in the mechanism. NCX function and SERCA activity are increased by apelin, and the SR Ca2+ content is decreased by apelin during twitches. PKC played an important role in these signaling mechanisms. PMID- 18424642 TI - Semiquantitative human cerebral perfusion assessment with ultrasound in brain space-occupying lesions: preliminary data. AB - OBJECTIVE: Transcranial Duplex ultrasound imaging with ultrasound contrast agents is an emerging technique for evaluating brain perfusion. The aim of this study was to evaluate cerebral perfusion with ultrasound in brain space-occupying lesions to identify different perfusion patterns. METHODS: Twenty patients with brain space-occupying lesions underwent ultrasound assessment of brain perfusion with a contrast pulse sequencing nonharmonic ultrasound technique and an ultrasound contrast agent bolus. Data were analyzed with software for semiquantitative analysis. RESULTS: Contrast pulse sequencing imaging with the semiquantitative analysis software allowed identification of qualitative and semiquantitative brain perfusion. Brain hemorrhages showed lower or absent perfusion compared with normal tissue. Meningiomas and glioblastomas without large necrotic areas showed higher perfusion compared with normal tissue. Glioblastomas with large necrotic areas showed overall reduced perfusion compared with normal tissue but higher than that of brain hemorrhages. In glioblastomas with large necrotic areas, it was possible to distinguish between solid and necrotic tissue. CONCLUSIONS: This bedside ultrasound technique, if validated by larger-scale studies, may add helpful information in noninvasive staging of brain tumors. Further potential applications may be in follow-up imaging to evaluate postoperative tumor recurrence or the presence of radionecrosis. PMID- 18424643 TI - Brachial artery pulsatility index change 1 minute after 5-minute forearm compression: comparison with flow-mediated dilatation. AB - OBJECTIVE: Endothelial impairment evaluation by sonographic measurement of flow mediated dilatation (FMD) has become broadly used. However, this method has 2 main caveats: the dilatation depends on the baseline arterial diameter, and a high precision level is required. Vasodilatation leads to an amplified fall in impedance. We hypothesized that assessment of the pulsatility index change (PI-C) 1 minute after 5-minute forearm compression might evaluate that fall in impedance. The aim of this study was to compare the PI-C with FMD. METHODS: Flow mediated dilatation and the PI-C were assessed in 51 healthy women aged between 35.1 and 67.1 years. We correlated both FMD and the PI-C with age, body mass index, waist circumference, cholesterol level, high-density lipoprotein level, glucose level, systolic and diastolic blood pressure, pulse pressure, brachial artery diameter, simplified Framingham score, intima-media thickness, and carotid stiffness index. Intraclass correlation coefficients between 2 FMD and PI-C measurements were also examined. RESULTS: Only FMD correlated with baseline brachial diameter (r = -0.53). The PI-C had a high correlation with age, body mass index, waist circumference, cholesterol level, systolic blood pressure, pulse pressure, simplified Framingham score, and intima-media thickness. The correlation between FMD and the PI-C was high (r = -0.66). The PI-C had a higher intraclass correlation coefficient (0.991) than FMD (0.836) but not brachial artery diameter (0.989). CONCLUSIONS: The PI-C had a large correlation with various markers of cardiovascular risk. Additionally, PI-C measurement does not require offline analysis, extra software, or electrocardiography. We think that the PI-C could be considered a marker of endothelial function. However, more studies are required before further conclusions. PMID- 18424644 TI - Sonographic nomogram of the fetal calf between 15 and 42 weeks' gestation using 3 dimensional sonography. AB - OBJECTIVE: The purpose of this study was to establish a new reference growth chart of calf muscle biometric measurements throughout gestation in normal singleton pregnancies. METHODS: A prospective cohort study was designed. One hundred pregnant women were included in the study and assessed by 3-dimensional sonography. Excluded were those with multiple pregnancies, congenital anomalies, abnormal karyotypes, and polyhydramnios or oligohydramnios. Three-dimensional multiplanar sonographic images were used to measure the calf muscles: soleus, gastrocnemius, popliteal, peroneus longus, and tibialis posterior. RESULTS: Calf muscle widths increased with increasing gestational age (R(2) = 0.857; P < .0001), and the ratio between calf muscles and the tibia increased as well (R(2) = 0.356; P = .001). CONCLUSIONS: The data provide a nomogram of calf width with a direct correlation between muscle growth and gestational age. This nomogram can offer a basis for normal calf development and may assist in distinguishing between different etiologies leading to clubfoot and other joint contractures associated with calf atrophy. PMID- 18424645 TI - The lung-to-thorax transverse area ratio at term and near term correlates with survival in isolated congenital diaphragmatic hernia. AB - OBJECTIVE: The purpose of this study was to determine how well liver position, the lung area-to-head circumference (L/H) ratio, and the lung-to-thorax transverse area (L/T) ratio predicted the need for extra-corporeal membrane oxygenation (ECMO) and survival in fetuses with isolated congenital diaphragmatic hernia (CDH). METHODS: Antenatal records of 25 fetuses with isolated left-sided CDH who were born by cesarean delivery under fetal stabilization at this institution were reviewed. The latest determinations of the L/H and L/T ratios before birth (between 34 and 38 weeks' gestation) were compared on the basis of the cutoff points for mortality: less than 1.0 versus 1.0 or greater for the L/H ratio and 0.08 or less versus greater than 0.08 for the L/T ratio. Outcome measures assessed were survival (discharge to home) and the need for ECMO. RESULTS: Overall survival was 64% (16/25). Postnatal survival in fetuses with an L/T ratio of 0.08 or less was statistically lower than in those with an L/T ratio of greater than 0.08 (33% versus 81%; P = .0308). The percentage requiring ECMO in the group with an L/T ratio of 0.08 or less was also higher than that of the group with an L/T ratio of greater than 0.08, but the difference was not statistically significant (67% versus 25%; P = .0872). Neither the L/H ratio nor herniation of the fetal liver into the chest affected survival or the need for ECMO. CONCLUSIONS: In fetuses with isolated CDH at term or near term, the L/T ratio may be a better predictor of outcome than the L/H ratio or liver herniation. PMID- 18424646 TI - Prenatal course of isolated muscular ventricular septal defects diagnosed only by color Doppler sonography: single-institution experience. AB - OBJECTIVE: Counseling patients with an isolated ventricular septal defect (i-VSD) is clinically important because with high-resolution ultrasound equipment, more small muscular VSDs are now being diagnosed. The prevalence of these lesions is not yet completely described, and the frequency with which muscular VSDs resolve in utero has also not been extensively reported. METHODS: We investigated the perinatal course of isolated muscular VSDs diagnosed only on color Doppler examinations and followed between January 1, 2005, and December 31, 2006. A complete evaluation of the fetal heart was performed by gray scale, spectral Doppler, and color Doppler examinations. RESULTS: We performed a total of 2583 fetal echocardiographic examinations on 2410 fetuses during 2318 pregnancies. The study group included 78 twin gestations (3.4%) and 7 triplet gestations (0.3%). There were 16 fetuses with an i-VSD (6.6/1000 fetuses) within the study group. The mean gestational age +/- SD at diagnosis was 23.5 +/- 4.3 weeks. Two of the i VSDs (12.5%) spontaneously resolved prenatally. One fetus with an i-VSD had trisomy 21 and also had increased nuchal translucency in the first trimester. One i-VSD was diagnosed among 22 fetuses with trisomy 21 examined during the study period. CONCLUSIONS: An i-VSD is a common congenital heart defect. Prenatal resolution of i-VSDs is less frequent than reported in the literature. A larger cohort is needed to provide a better risk estimate for aneuploidy in the presence of an i-VSD. PMID- 18424647 TI - Assessment of factors that affect the quality of performance and interpretation of sonography of adnexal masses. AB - OBJECTIVE: The purpose of this study was to assess factors that affect the quality of performance and interpretation of sonography of adnexal masses. METHODS: Two gynecologic sonographers performed blinded reviews of up to 3 sonograms within 5 years before surgery in 325 consecutive women who underwent oophorectomy (610 sonograms). Three 5-point quality scores were assigned (with 5 being the best score) on the basis of the technical quality of the images, accurate description of findings, and summary impression in the report. The location of the examination (on-site, remote, or emergency department), type of fellowship, practice experience (<5, 5-10, or >10 years), and specialty (women's imaging, abdominal imaging, or other) of the radiologists were recorded. Analysis of variance was used to assess the impact of these multiple factors on quality. RESULTS: No significant differences were found among 31 radiologists on the basis of the type of training, years in practice, or number of examinations read. Average scores among radiologists for technical quality, findings, and impressions were 4.96, 4.88, and 4.83, respectively. Radiologists who specialized in women's imaging performed best in the quality of their impressions (4.86 versus 4.79; P = .029). There were no significant differences in scores with respect to the examination location. CONCLUSIONS: Within this sample of radiologists, the technical quality of the examinations was not affected by the type of training or subspecialty practice. Reports of adnexal mass findings were accurately described by all radiologists, although specialization in women's imaging improved the ability to provide an accurate impression. PMID- 18424648 TI - Three-dimensional ultrasonographic application for analyzing synovial hypertrophy of the knee in patients with osteoarthritis. AB - OBJECTIVE: The purpose of this study was to evaluate 3-dimensional (3D) ultrasonography for the visualization of intra-articular synovial hypertrophy in patients with osteoarthritis. METHODS: Knee joints of 22 patients with osteoarthritis were examined by 3D ultrasonography, and their synovial fluids were analyzed. Ultrasonographic image patterns, vascular endothelial growth factor and transforming growth factor beta concentrations in synovial fluid, and serum inflammatory markers were analyzed. RESULTS: It was possible to visualize the intra-articular structure by 3D ultrasonography. Three-dimensional ultrasonography revealed various interesting stereoscopic views of the synovial structures, and the patterns of synovial proliferation ranged from simple proliferations to complex shrubby structures. Patients with a more complex and proliferative pattern in the synovium tended to have higher C-reactive protein concentrations, but this difference was not significant (P = .09). The concentrations of vascular endothelial growth factor and transforming growth factor beta in synovial fluid were significantly higher in patients with complex hypertrophy (P < .05). CONCLUSIONS: Three-dimensional technology was useful in delineating the shape of the synovium. It may have a possible impact on future imaging in rheumatology. PMID- 18424649 TI - Ultrasonographic measurements of the ulnar nerve at the elbow: role of confounders. AB - OBJECTIVE: The purpose of this study was to identify factors confounding high resolution ultrasonographic measurements of the ulnar nerve to test their influence when discriminating between limbs affected and unaffected by ulnar nerve entrapment (UNE) at the elbow. METHODS: High-resolution ultrasonographic measurements of ulnar nerve dimensions at the elbow were compared between 2 groups of subjects: symptomatic and asymptomatic for UNE. Rank analysis of covariance regression tests were performed to determine whether significant differences existed between the 2 groups. The changing coefficient method (using rank analysis of covariance tests) was used to test for potential confounding effects of age, weight, height, body mass index, sex, limb sidedness, limb handedness, and nerve mobility. These tests were repeated for each measurement while controlling for the identified confounders. Exact 2-tailed Wilcoxon signed rank tests were performed to test for significant differences between measurements of the diameter of the ulnar nerve with the elbow in full extension and full flexion. RESULTS: Age, weight, body mass index, sex, and elbow position were shown to have confounding influences on high-resolution ultrasonographic measurements of the ulnar nerve. No confounding effect was apparent for limb sidedness or dominance. Cross-sectional area and long-axis diameter measurements demonstrated significant differences between nerves with and without UNE after controlling for confounders. CONCLUSIONS: Two cross-sectional measurements (area and maximum cross-sectional diameter) of the ulnar nerve, made at the level of the medial epicondyle, were found to be robust discriminators between nerves with and without UNE. In the absence of normative reference values of the ulnar nerve, the contralateral limb may be used as the comparative control. PMID- 18424650 TI - A pilot study of comprehensive ultrasound education at the Wayne State University School of Medicine: a pioneer year review. AB - OBJECTIVE: Ultrasound is a versatile diagnostic modality used in a variety of medical fields. Wayne State University School of Medicine (WSUSOM) is one of the first medical schools in the United States to integrate an ultrasound curriculum through both basic science courses and clinical clerkships. METHODS: In 2006, 25 portable ultrasound units were donated to WSUSOM. First-year medical students were provided an ultrasound curriculum consisting of 6 organ-system sessions that addressed the basics of ultrasound techniques, anatomy, and procedural skills. After the last session, students were administered 2 anonymous and voluntary evaluations. The first assessed their overall experience with the ultrasound curriculum, and the second assessed their technical skills in applying ultrasound techniques. RESULTS: Eighty-three percent of students agreed or strongly agreed that their experience with ultrasound education was positive. On the summative evaluation, nearly 91% of students agreed or strongly agreed that they would benefit from continued ultrasound education throughout their 4 years of medical school. Student performance on the technical assessment was also very positive, with mean class performance of 87%. CONCLUSIONS: As residency programs adopt ultrasound training, medical school faculty should consider incorporating ultrasound education into their curriculum. Portable ultrasound has the potential to be used in many different settings, including rural practice sites and sporting events. The WSUSOM committee's pilot ultrasound curriculum will continue to use student feedback to enhance the ultrasound experience, helping students prepare for challenges that they will face in the future. PMID- 18424651 TI - Hurthle cell neoplasms of the thyroid: sonographic appearance and histologic characteristics. AB - OBJECTIVE: The purpose of this study was to determine the sonographic features of Hurthle cell neoplasms (HCNs) of the thyroid. METHODS: We retrospectively analyzed the sonographic appearance of 15 histologically proven HCNs in 15 patients aged 16 to 70 years (mean age, 44 years). Sonographic features that were reviewed included the size and echogenicity of the tumors, the presence of cystic areas or calcifications, and detectable blood flow on color Doppler imaging. Correlation of sonographic findings with pathologic results was performed. RESULTS: The tumors ranged from 0.4 to 7 cm in diameter, but most were less than 3 cm in diameter. Four (27%) of the 15 tumors were homogeneously hypoechoic. Two tumors (13%) were predominantly hypoechoic with isoechoic areas to thyroid parenchyma. Two (13%) neoplasms were isoechoic to thyroid parenchyma. Four (27%) tumors were predominantly isoechoic, containing hypoechoic areas, and 3 (20%) tumors were hyperechoic. Three neoplasms contained cystic components. None of the tumors contained calcifications. One tumor was avascular on Doppler examination. One neoplasm showed only peripheral blood flow. Thirteen tumors showed internal vascularity, 7 of them with peripheral blood flow. Twelve HCNs were benign, and 3 were malignant on pathologic examination. CONCLUSIONS: Hurthle cell neoplasms show a spectrum of sonographic appearances from predominantly hypoechoic to hyperechoic lesions and from peripheral blood flow with no internal flow to extensively vascularized lesions. Pathologic criteria differentiating benign and malignant HCNs (absence or presence of a capsular breach, vascular or extrathyroidal tissue invasion, nodal involvement, and distant metastasis) are beyond the resolution of sonography and fine-needle aspiration biopsy and require removal of the entire lesion. This precludes diagnosis and characterization of HCNs by sonography. PMID- 18424652 TI - Sonography of the ear pinna. AB - OBJECTIVE: This study was undertaken to assess the scope of sonography for the diagnosis of diseases of the external ear. METHODS: We developed a description of the sonographic anatomy of the normal ear pinna, including the thickness of the normal cartilage and lobule, through systematic scanning of the external ears of 11 healthy volunteers (2 male and 9 female), and reviewed clinical cases with pathologic entities. RESULTS: Reproducible and recognizable images were collected from normal as well as pathologic cases of the external ear. Images are presented for reference. CONCLUSIONS: Sonography permits good visualization of the internal structure, including the cartilage, and it is possible to use sonography as a method of study to differentiate between inflammation, vascular lesions, and tumors. PMID- 18424653 TI - Application of automated sonography on 4-dimensional volumes of fetuses with transposition of the great arteries. AB - OBJECTIVE: The purpose of this study was to assess, in second-trimester fetuses with transposition of the great arteries (TGA), the performance of software (sonographically based volume computer-aided analysis) that automatically retrieves diagnostic cardiac planes from a 4-dimensional volume of the fetal chest obtained with spatiotemporal image correlation. METHODS: We retrospectively evaluated the 4-dimensional spatiotemporal image correlation volumes of 12 fetuses with TGA (complete TGA, 10 cases; correct TGA, 2 cases). The data were analyzed to determine whether the target diagnostic planes, that is, cardiac plane 1 (left ventricle outflow tract) and cardiac plane 2 (right ventricle outflow tract), were correctly identified in at least 1 of the 7 automatically generated tomographic sonographic image displays and whether they allowed diagnosis of TGA. RESULTS: In 9 of 10 fetuses with complete TGA, target diagnostic cardiac plane 1 showed a branching arterial vessel (pulmonary artery) arising from the left ventricle, whereas in 7 of 10 fetuses, the aorta arising from the right ventricle was shown. In both cases with correct TGA, the pulmonary artery starting from the morphologic left ventricle was shown, whereas in 1 of 2, the connection of the aorta with the morphologic right ventricle was found. In all of the fetuses with TGA, a ventricular arterial connection anomaly was shown in either cardiac plane 1 or 2. CONCLUSIONS: This automatic approach shows good retrieval of diagnostic cardiac planes in fetuses with TGA, which may improve diagnostic efficacy for this disease. PMID- 18424654 TI - Role of sonography in the recognition, assessment, and treatment of cesarean scar ectopic pregnancies. AB - OBJECTIVE: Cesarean scar ectopic pregnancies (CSEPs) are rare but may have serious adverse consequences and are therefore important to promptly recognize on sonography. We aim to describe the typical sonographic appearances. Potential treatments are discussed, including sonographic guidance for transcervical injection of methotrexate (MTX) into the gestational sac. METHODS: Two patients with CSEPs were treated with systemic and intra-amniotic administration of MTX under sonographic guidance. RESULTS: Both patients were followed clinically after medical treatment, resulting in low maternal morbidity and mortality. CONCLUSIONS: Considering the increasing rate of cesarean delivery and the increased risk of CSEPs, sonologists should be familiar with the sonographic appearances of a pregnancy implanted into the cesarean scar. We show how to correctly diagnose scar implantation and describe how to perform sonographically guided transcervical injection of MTX. PMID- 18424655 TI - Point-of-care sonographic detection of left endobronchial main stem intubation and obstruction versus endotracheal intubation. AB - OBJECTIVE: Determining the correct position of endotracheal tubes in critically ill patients may be complicated by external factors such as noise, body habitus, and the need for ongoing resuscitation. Multiple detection techniques have been developed to determine the correct endotracheal tube position, recently including the use of sonography to evaluate lung expansion and diaphragmatic excursion. These techniques have also been applied to diagnosis of right endobronchial main stem intubation, which may be confused with a unilateral pneumothorax in some cases. METHODS: We describe the sonographic findings in a case series of endobronchial main stem intubations and obstruction, highlighting the utility of this sonographic application. Previous literature and future applications are discussed. RESULTS: Sonographic detection of the sliding lung sign, the lung pulse, and diaphragmatic excursion can accurately detect main stem bronchial intubation as well as bronchial obstruction. CONCLUSIONS: Clinical use of lung sonography may decrease the need for chest radiography and may allow more rapid diagnosis of main stem intubation and bronchial obstruction. PMID- 18424656 TI - Ultrasonographic cerebral perfusion in assessment of brain death: a preliminary study. AB - OBJECTIVE: The purpose of this series was to describe the ultrasonographic perfusion pattern in patients with brain death. METHODS: Thirteen patients with different neurologic disorders in whom brain death developed were studied. Transcranial perfusion was analyzed after injection of 2.5 mL of a sulfur hexafluoride ultrasonographic contrast agent. Time-intensity curves were analyzed in predetermined regions of interest. RESULTS: In all patients, analysis of regions of interest showed no bolus-like curve progression. This finding implies a complete absence of cerebral perfusion. CONCLUSIONS: Patients with brain death studied by ultrasonographic perfusion techniques have a characteristic pattern. PMID- 18424657 TI - Intracranial dissection and extracranial hypoplasia of the internal carotid artery. PMID- 18424658 TI - Congenital dermal sinus. PMID- 18424659 TI - Inflammatory pseudotumor of the kidney. PMID- 18424660 TI - Sonographic features of an ossifying fibromyxoid tumor of the buttock. PMID- 18424661 TI - Sonographic findings of an eccrine spiradenoma: case report and literature review. PMID- 18424662 TI - Mannosylated self-peptide inhibits the development of experimental autoimmune encephalomyelitis via expansion of nonencephalitogenic T cells. AB - Tolerance to experimental autoimmune encephalomyelitis (EAE) in SJL mice can be induced by immunization with a mannosylated form of the proteolipid protein (M PLP139-151), despite the presence of CFA. The state of tolerance is characterized by poor delayed-type hypersensitivity responses and the absence of clinical EAE symptoms. In vivo monitoring of CFSE-labeled PLP139-151-specific TCR-transgenic (5B6) T cells revealed that immunization with M-PLP139-151 increases the clonal expansion of 5B6 T cells that do not develop full effector functions. Moreover, nonfunctional T cells obtained from M-PLP139-151-immunized mice showed poor blastogenesis and were unable to transfer EAE to naive recipients. Nevertheless, the in vitro production of cytokines and chemokines associated with EAE was unaffected. Importantly, tolerance induced by M-PLP139-151 was abrogated by the administration of pertussis toxin, resulting in EAE development. Our results suggest that M-PLP139-151 inhibits EAE development by affecting the differentiation of T cells into encephalitogenic effector cells. PMID- 18424663 TI - Human mast cell activation with virus-associated stimuli leads to the selective chemotaxis of natural killer cells by a CXCL8-dependent mechanism. AB - Human mast cells are found in skin and mucosal surfaces and next to blood vessels. They play a sentinel cell role in immunity, recognizing invading pathogens and producing proinflammatory mediators. Mast cells can recruit granulocytes, and monocytes in allergic disease and bacterial infection, but their ability to recruit antiviral effector cells such as natural killer (NK) cells and T cells has not been fully elucidated. To investigate the role of human mast cells in response to virus-associated stimuli, human cord blood-derived mast cells (CBMCs) were stimulated with polyinosinic.polycytidylic acid, a double stranded RNA analog, or infected with the double-stranded RNA virus, reovirus serotype 3 Dearing for 24 hours. CBMCs responded to stimulation with polyinosinic.polycytidylic acid by producing a distinct chemokine profile, including CCL4, CXCL8, and CXCL10. CBMCs produced significant amounts of CXCL8 in response to low levels of reovirus infection, while both skin- and lung-derived fibroblasts were unresponsive unless higher doses of reovirus were used. Supernatants from CBMCs infected with reovirus induced substantial NK cell chemotaxis that was highly dependent on CXCL8 and CXCR1. These results suggest a novel role for mast cells in the recruitment of human NK cells to sites of early viral infection via CXCL8. PMID- 18424664 TI - CD38 gene polymorphism and chronic lymphocytic leukemia: a role in transformation to Richter syndrome? AB - CD38 rules proliferation signals in chronic lymphocytic leukemia (CLL) cells, suggesting that the molecule is not merely a prognostic marker but also a key element in the pathogenetic network underlying the disease. CD38 has a genetic polymorphism, characterized by a C>G variation in the regulatory region of intron 1. The working hypothesis is that the presence of different alleles in CLL patients marks (or accounts for) some of the clinical heterogeneity. CD38 allele distribution in 248 Italian patients overlapped with that of the controls (n = 232), suggesting that susceptibility to CLL is not influenced by CD38 genotype. Stratification of patients according to markers of unfavorable prognosis constantly resulted in a significantly higher frequency of the rare G allele. Furthermore, analysis of clinical parameters showed that G allele is independently associated with nodal/splenic involvement. The highest G allele frequency was observed in the 16 patients of the cohort that developed Richter syndrome (RS). Five-year cumulative incidence of transformation was significantly higher in G allele carriers than in CC homozygotes. Multivariate analysis on a total of 30 RS patients confirmed that the probability of transformation is strongly associated with G allele, likely representing an independent risk factor for RS development. PMID- 18424665 TI - Flk2+ common lymphoid progenitors possess equivalent differentiation potential for the B and T lineages. AB - Mature blood cells develop from multipotent hematopoietic stem cells through a series of sequential intermediates in which the developmental potential for particular blood lineages is progressively extinguished. We previously reported the identification of one of these developmental intermediates, the common lymphoid progenitor (CLP), which can give rise to T cells, B cells, dendritic cells (DCs), and natural killer cells (NKs), but lacks myeloid and erythroid potential. Recently, several studies have suggested that the T-cell and DC potential of CLP is limited or absent, and/or that CLP contains significant myeloid potential. Here, we show that the originally identified CLP population can be divided into functionally distinct subsets based on the expression of the tyrosine kinase receptor, Flk2. The Flk2(+) subset contains robust in vivo and in vitro T-cell, B-cell, DC, and NK potential, but lacks myeloid potential and, therefore, represents an oligopotent, lymphoid-restricted progenitor. This population of cells does not appear to be B cell-biased and robustly reconstitutes both B and T lineages in vivo, consistent with its being a physiologic progenitor of both of these subsets. Thus, Flk2 expression defines a homogeneous, readily obtainable subset of bone marrow CLP that is completely lymphoid-committed and can differentiate equivalently well into both B and T lineages. PMID- 18424666 TI - Insight into the pathogenesis of chronic lymphocytic leukemia (CLL) through analysis of IgVH gene usage and mutation status in familial CLL. AB - To address the proposition that familial B-cell chronic lymphocytic leukemia (CLL) may exhibit a more restricted phenotype than sporadic CLL with respect to immunoglobulin gene usage or ontogenic development, we compared immunoglobulin (Ig) heavy chain variable region (VH) gene usage and IgVH mutation status in 327 patients with CLL from 214 families with 724 patients with sporadic cases. The frequency of mutated CLL was higher in familial CLL (P < .001), and there was evidence of intrafamilial concordance in mutation status (P < .001). The repertoire and frequency of IgVH usage was, however, not significantly different between familial and sporadic CLL. Furthermore, IgVH usage was not correlated between affected members of the same family. These observations provide evidence that familial CLL is essentially indistinguishable from sporadic CLL, favoring a genetic basis to disease development in general rather than a simple environmental etiology. PMID- 18424668 TI - Behavioral and neural responses of juvenile crayfish to moving shadows. AB - One of the most important decisions any animal has to make is how to respond to sensory cues that suggest an imminent attack by a predator. We measured behavioral and neural responses of juvenile crayfish to moving shadows of different velocities while the animals were searching for food. In all experiments, and independent of shadow velocity, each crayfish produced one of two discrete behavioral outputs: it either tail-flipped backwards by rapid flexion of its abdomen or it immediately stopped its forward locomotion. The probability of each behavioral response was dependent on the velocity of the shadows that were presented. While most animals responded with tail-flips to slow moving shadows and stops were rarely observed, the number of tail-flips decreased as shadow velocity increased. Tail-flips were almost absent for very fast-moving shadows and stopping behavior became the dominating response. By using a non invasive technique to record neural activity, we were able to identify the underlying neural circuit that controlled the observed tail-flips. All tail-flips were mediated by activation of the medial giant neurons, which are part of a hardwired neural circuit previously described to produce reflexive responses to tactile stimulation. PMID- 18424669 TI - Comparative sequence analysis of myosin heavy chain proteins from congeneric shallow- and deep-living rattail fish (genus Coryphaenoides). AB - The evolutionary adaptations of functional genes to life at high pressure are not well understood. To elucidate the mechanisms of protein adaptation to high pressure, we cloned the myosin heavy chain (MyHC) cDNA from skeletal muscle of two deep-sea fishes, Coryphaenoides yaquinae and C. armatus, and two non-deep-sea fishes, C. acrolepis and C. cinereus. The MyHCs of deep-sea fishes have a unique structure in two loop regions, loop-1 and loop-2, in comparison with those of non deep-sea fishes. The loop-1 region of deep-sea fishes has a Pro residue and the loop-2 region, which is an actin-binding site, is shorter than the same region in non-deep-sea fishes. The amino acid substitution in the loop-1 region is expected to be mainly involved in ATPase activity, whereas the deletion in the loop-2 region affects the association of MyHC with actin filaments at high pressure. In addition, the MyHC of deep-sea fishes has biased amino acid substitutions at core positions in the coiled-coil structure of the rod region. These amino acid substitutions are likely to decrease the cavities in the coiled-coil structure and consequently make the structure more compact and unaffected by high pressure. Together, these results indicate that amino acid substitutions can adaptively alter the pressure sensitivity of a protein even if they do not directly influence core structure. PMID- 18424667 TI - Borrelia infection and risk of non-Hodgkin lymphoma. AB - Reports of the presence of Borrelia burgdorferi DNA in malignant lymphomas have raised the hypothesis that infection with B. burgdorferi may be causally related to non-Hodgkin lymphoma (NHL) development. We conducted a Danish-Swedish case control study including 3055 NHL patients and 3187 population controls. History of tick bite or Borrelia infection was ascertained through structured telephone interviews and through enzyme-linked immunosorbent assay serum analyses for antibodies against B. burgdorferi in a subset of 1579 patients and 1358 controls. Statistical associations with risk of NHL, including histologic subtypes, were assessed by logistic regression. Overall risk of NHL was not associated with self reported history of tick bite (odds ratio [OR] = 1.0; 95% confidence interval: 0.9-1.1), Borrelia infection (OR = 1.3 [0.96-1.8]) or the presence of anti Borrelia antibodies (OR = 1.3 [0.9-2.0]). However, in analyses of NHL subtypes, self-reported history of B. burgdorferi infection (OR = 2.5 [1.2-5.1]) and seropositivity for anti-Borrelia antibodies (OR = 3.6 [1.8-7.4]) were both associated with risk of mantle cell lymphoma. Notably, this specific association was also observed in persons who did not recall Borrelia infection yet tested positive for anti-Borrelia antibodies (OR = 4.2 [2.0-8.9]). Our observations suggest a previously unreported association between B. burgdorferi infection and risk of mantle cell lymphoma. PMID- 18424670 TI - Lower-limb biomechanics during stair descent: influence of step-height and body mass. AB - The aim of the present study was to examine the biomechanics of the lower limb during stair descent and the effects of increasing demand in two ways: by increasing step-height and by increasing body mass. Ten male subjects walked down a four-step staircase, the height of which could be altered. The step-heights were: standard (17 cm), 50% decreased, 50% increased and 75% increased. At the standard height, subjects also walked down wearing a weighted jacket carrying 20% extra body mass. Lower limb kinematics and kinetics were determined using motion capture and ground reaction forces. Also measured were gastrocnemius medialis (GM) muscle electromyography and GM muscle fascicle length using ultrasonography. GM muscle fascicles actively shortened during the touch-down phase of stair descent in all conditions, while the muscle-tendon complex (MTC), as calculated from the knee and ankle joint kinematics, lengthened. The GM muscle fascicles shortened more when step-height was increased, which corresponded to the increase in ankle joint moment. Increased body mass did not alter the ankle or knee joint moment in the first contact phase of a step down; due to a change in strategy, the trailing leg, instead of the leading leg, supported the extra mass. Hence, the amount of GM muscle fascicle shortening, during the touch-down phase, also did not change with added body mass. Our results suggest that the increase in joint moments is related to the amount of fascicle shortening, which occurs whilst the MTC is lengthening, thereby stretching the elastic tendinous tissues. PMID- 18424671 TI - Retinal processing and opponent mechanisms mediating ultraviolet polarization sensitivity in rainbow trout (Oncorhynchus mykiss). AB - A number of teleost fishes have photoreceptor mechanisms to detect linearly polarized light. We studied the neuronal mechanism underlying this ability. It was found that a polarized signal could be detected in rainbow trout (Oncorhynchus mykiss) both in the electroretinogram (ERG) and in the compound action potential (CAP) measured in the optic nerve, indicating a strong retinal contribution to the processing of polarized light. The CAP recordings showed a W shaped sensitivity curve, with a peak at 0 degrees , 90 degrees and 180 degrees , consistent with processes for both vertical and horizontal orientation. By contrast, the ERG recordings reveal a more complex pattern. In addition to the peaks at 0 degrees , 90 degrees and 180 degrees , two additional peaks appeared at 45 degrees and 135 degrees . This result suggests a specialized contribution of the outer retina in the processing of polarized light. The spectral sensitivity of the mechanisms responsible for these intermediate peaks was studied using chromatic adaptation. Here we show that long wavelength-sensitive (LWS) cone mechanism adaptation shifted the intermediate peaks towards 90 degrees , whereas ultraviolet-sensitive (UVS) cone mechanism adaptation shifted the peaks away from 90 degrees towards either 0 degrees or 180 degrees . These results provide further confirmation that the 90 degrees peak is dominated by the LWS cone mechanism and the 0 degrees and 180 degrees peaks are dominated by the UVS cone mechanism. In addition, a pharmacological approach was used to examine the retinal neural mechanisms underlying polarization sensitivity. The effect of blocking negative feedback from horizontal cells to cones on the ERG was studied by making intraocular injections of low doses of cobalt, known to block this feedback pathway. It was found that the intermediate peaks seen in the ERG polarization sensitivity curves were eliminated after application of cobalt, suggesting that these peaks are due to outer retinal inhibition derived from feedback of horizontal cells onto cones. A simple computational model was developed to evaluate these results. The model consists of opponent and non opponent processing elements for the two polarization detectors. This model provides a first approximation analysis suggesting that opponent processing occurs in the outer retina for polarization vision. Although it seems that polarization vision uses a slightly more complicated coding scheme than colour vision, the results presented in this paper suggest that opponent and non opponent channels process polarization information. PMID- 18424672 TI - The effects of acute temperature change on swimming performance in bluegill sunfish Lepomis macrochirus. AB - Many fish change gait within their aerobically supported range of swimming speeds. The effects of acute temperature change on this type of locomotor behavior are poorly understood. Bluegill sunfish swim in the labriform mode at low speeds and switch to undulatory swimming as their swimming speed increases. Maximum aerobic swimming speed (U(max)), labriform-undulatory gait transition speed (U(trans)) and the relationships between fin beat frequency and speed were measured at 14, 18, 22, 26 and 30 degrees C in bluegill acclimated to 22 degrees C. At temperatures below the acclimation temperature (T(a)), U(max), U(trans) and the caudal and pectoral fin beat frequencies at these speeds were reduced relative to the acclimation level. At temperatures above T(a) there was no change in these variables relative to the acclimation level. Supplementation of oxygen levels at 30 degrees C had no effect on swimming performance. The mechanical power output of the abductor superficialis, a pectoral fin abductor muscle, was measured in vitro at the same temperatures used for the swimming experiments. At and below T(a), maximal power output was produced at a cycle frequency approximately matching the in vivo pectoral fin beat frequency. At temperatures above T(a) muscle power output and cycle frequency could be increased above the in vivo levels at U(trans). Our data suggest that the factors triggering the labriform-undulatory gait transition change with temperature. Muscle mechanical performance limited labriform swimming speed at T(a) and below, but other mechanical or energetic factors limited labriform swimming speed at temperatures above T(a). PMID- 18424673 TI - Is there life in the horny layer? Dihydropyridine and ryanodine receptors in the skin of female and male chickens (Gallus domesticus). AB - Previous findings in pigeons and chickens show that Ca(2+) may be accumulated inside the cornified skin cells and that Ca(2+) microenvironments with a lower- or higher-than-blood concentration may exist in the skin. It has been suggested that the skin may function as a secretory pathway or a reservoir for Ca(2+) recycling. To test this hypothesis, we studied the dermis and epidermis of female and male chickens in vivo to find out whether cellular mechanisms exist for the accumulation, recycling or secretion of Ca(2+). For calcium influx and intracellular Ca(2+) release, respectively, the density of dihydropyridine receptors (DHPRs) and ryanodine receptors (RyRs) was examined, using high affinity (-)-enantiomers of dihydropyridine and ryanodine labelled with fluorophores. To investigate Ca(2+) utilization in the skin, the systemic and local activity of the enzyme alkaline phosphatase (ALP) and the concentration of ionic Ca(2+) were measured in plasma and in cutaneous extracellular fluid, collected by suction blister technique. We found that both DHPRs and RyRs were present in all skin layers from dermis to horny layer. However, receptor densities were highest in the surface layers. With a basic calcium-rich diet, receptor densities were higher in males, particularly in the dermis and mid epidermis. After a reduction in the nutritional Ca(2+) input, receptor densities in males decreased to the same level as in females, in which the receptor densities were not affected by the amount of Ca(2+) in the diet or that resulting from coming out of lay. The extracellular concentration of ionic Ca(2+) per se was not found to affect the density of DHPRs and RyRs in the skin. Spatially, RyRs seem to be located in the periphery of the sebokeratinocyte. ALP activity was shown to be lower in the extracellular fluid than in the plasma in both sexes. However, activity in both extracellular domains increased significantly in females that had come out of lay. This was probably connected with the increased osteoblast activity related to the reformation of structural bone. In conclusion, voltage-sensitive L-type Ca(2+) channels for ion influx and RyRs for Ca(2+) release are present in the cells of the skin of female and male chickens. Higher densities in the males receiving excessive Ca(2+) imply an increased capacity for Ca(2+) influx and intracellular processing. Even though the functional interactions between DHPRs and RyRs in the sebokeratinocytes could not be demonstrated, peripheral colocation and high receptor densities at the level of exocytosis of the lamellar bodies point to their role as part of a signalling pathway for secretion. The finding that DHPRs and RyRs are present in the horny layer implies that the function of the outermost skin might be more active than had been previously thought and that this function might be both secretory and sensory. PMID- 18424674 TI - Mechanics and energetics of level walking with powered ankle exoskeletons. AB - Robotic lower limb exoskeletons that can alter joint mechanical power output are novel tools for studying the relationship between the mechanics and energetics of human locomotion. We built pneumatically powered ankle exoskeletons controlled by the user's own soleus electromyography (i.e. proportional myoelectric control) to determine whether mechanical assistance at the ankle joint could reduce the metabolic cost of level, steady-speed human walking. We hypothesized that subjects would reduce their net metabolic power in proportion to the average positive mechanical power delivered by the bilateral ankle exoskeletons. Nine healthy individuals completed three 30 min sessions walking at 1.25 m s(-1) while wearing the exoskeletons. Over the three sessions, subjects' net metabolic energy expenditure during powered walking progressed from +7% to -10% of that during unpowered walking. With practice, subjects significantly reduced soleus muscle activity (by approximately 28% root mean square EMG, P<0.0001) and negative exoskeleton mechanical power (-0.09 W kg(-1) at the beginning of session 1 and 0.03 W kg(-1) at the end of session 3; P=0.005). Ankle joint kinematics returned to similar patterns to those observed during unpowered walking. At the end of the third session, the powered exoskeletons delivered approximately 63% of the average ankle joint positive mechanical power and approximately 22% of the total positive mechanical power generated by all of the joints summed (ankle, knee and hip) during unpowered walking. Decreases in total joint positive mechanical power due to powered ankle assistance ( approximately 22%) were not proportional to reductions in net metabolic power ( approximately 10%). The ;apparent efficiency' of the ankle joint muscle-tendon system during human walking ( approximately 0.61) was much greater than reported values of the ;muscular efficiency' of positive mechanical work for human muscle ( approximately 0.10-0.34). High ankle joint ;apparent efficiency' suggests that recoiling Achilles' tendon contributes a significant amount of ankle joint positive power during the push-off phase of walking in humans. PMID- 18424675 TI - Effects of maternal carotenoid availability in relation to sex, parasite infection and health status of nestling kestrels (Falco tinnunculus). AB - Mothers can strongly influence the development of their offspring, and if maternal resources are limited, they may influence optimal reproductive strategies. In birds, maternally deposited carotenoids are a prominent component of egg yolk and are vital for the development of the embryo. However, results of long-lasting fitness consequences of this early nutritional environment have been scarce and inconsistent. In addition, sex-biased sensitivity to different egg components is one of the mechanisms postulated to account for sex-linked environmental vulnerability during early life. However, this important aspect is usually not accounted for when investigating maternal investment in carotenoids. In this study we gave carotenoid (lutein) supplements to female Eurasian kestrels (Falco tinnunculus) before and during egg laying. The experiment increased female plasma carotenoids, but this effect was not apparent in hatchling and fledgling plasma carotenoid concentration. Also, results showed that carotenoid supplementation increased the high density lipoprotein to low density lipoprotein ratio in adult females, suggesting that dietary carotenoids may influence lipid metabolism. Furthermore, the effect of the treatment was manifested in several nestling health state parameters. Nestlings of carotenoid-supplemented females were infested by less intestinal parasite groups, had higher lymphocyte concentrations in blood plasma, and were less stressed (heterophile to lymphocyte ratio) than control nestlings. In addition, an interaction between the experimental treatment and nestling sex was apparent for globulin concentrations, favouring the smaller male nestlings. Thereby, suggesting that males benefited more than females from an increase in maternal carotenoid investment. Our study shows that an increase in carotenoids in the maternal diet during egg laying favours nestling development in kestrels, and may also affect nestlings in a sex specific way. PMID- 18424676 TI - Aldehyde-encapsulating liposomes impair marine grazer survivorship. AB - In the last decade, there has been an increased awareness that secondary metabolites produced by marine diatoms negatively impact the reproductive success of their principal predators, the copepods. Several oxylipins, products of the enzymatic oxidation of fatty acids, are produced when these unicellular algae are damaged, as occurs during grazing. In the past, the dinoflagellate Prorocentrum minimum, which does not produce the oxylipin 2-trans,4-trans-decadienal (DD), has been used as a live carrier to calculate daily ingestion rates of this molecule by copepod crustaceans. However, since the interaction between oxylipins and live carriers is unknown, the question as to how much and for how long ingestion of these molecules affects copepod reproduction remains a critical point to understanding the functional role of such compounds at sea. In the investigation presented here we used giant liposomes ( approximately 7 mum) as a delivery system for the oxylipin DD, prepared in the same size range as copepod food and containing known amounts of DD. The aim of this work was to relate the ingestion of DD to the reproductive failure of the copepods Temora stylifera and Calanus helgolandicus. Liposomes were very stable over time and after 10 days of feeding, liposomes encapsulating DD reduced egg hatching success and female survival with a concomitant appearance of apoptosis in both copepod embryos and female tissues. Concentrations of DD inducing blockage were one order of magnitude lower that those used in classical feeding experiments demonstrating that liposomes are a useful tool to quantitatively analyze the impact of toxins on copepods. PMID- 18424677 TI - The pyloric neural circuit of the herbivorous crab Pugettia producta shows limited sensitivity to several neuromodulators that elicit robust effects in more opportunistically feeding decapods. AB - Modulation of neural circuits in the crustacean stomatogastric nervous system (STNS) allows flexibility in the movements of the foregut musculature. The extensive repertoire of such resulting motor patterns in dietary generalists is hypothesized to permit these animals to process varied foods. The foregut and STNS of Pugettia producta are similar to those of other decapods, but its diet is more uniform, consisting primarily of kelp. We investigated the distribution of highly conserved neuromodulators in the stomatogastric ganglion (STG) and neuroendocrine organs of Pugettia, and documented their effects on its pyloric rhythm. Using immunohistochemistry, we found that the distributions of Cancer borealis tachykinin-related peptide I (CabTRP I), crustacean cardioactive peptide (CCAP), proctolin, red pigment concentrating hormone (RPCH) and tyrosine hydroxylase (dopamine) were similar to those of other decapods. For all peptides except proctolin, the isoforms responsible for the immunoreactivity were confirmed by mass spectrometry to be the authentic peptides. Only two modulators had physiological effects on the pyloric circuit similar to those seen in other species. In non-rhythmic preparations, proctolin and the muscarinic acetylcholine agonist oxotremorine consistently initiated a full pyloric rhythm. Dopamine usually activated a pyloric rhythm, but this pattern was highly variable. In only about 25% of preparations, RPCH activated a pyloric rhythm similar to that seen in other species. CCAP and CabTRP I had no effect on the pyloric rhythm. Thus, whereas Pugettia possesses all the neuromodulators investigated, its pyloric rhythm, when compared with other decapods, appears less sensitive to many of them, perhaps because of its limited diet. PMID- 18424678 TI - Role of the PGC-1 family in the metabolic adaptation of goldfish to diet and temperature. AB - In mammals, the peroxisome proliferator-activated receptor (PPAR) gamma coactivator-1 (PGC-1) family members and their binding partners orchestrate remodelling in response to diverse challenges such as diet, temperature and exercise. In this study, we exposed goldfish to three temperatures (4, 20 and 35 degrees C) and to three dietary regimes (food deprivation, low fat and high fat) and examined the changes in mitochondrial enzyme activities and transcript levels for metabolic enzymes and their genetic regulators in red muscle, white muscle, heart and liver. When all tissues and conditions were pooled, there were significant correlations between the mRNA for the PGC-1 coactivators (both alpha and beta) and mitochondrial transcripts (citrate synthase), metabolic gene regulators including PPARalpha, PPARbeta and nuclear respiratory factor-1 (NRF 1). PGC-1beta was the better predictor of the NRF-1 axis, whereas PGC-1alpha was the better predictor of the PPAR axis (PPARalpha, PPARbeta, medium chain acyl CoA dehydrogenase). In contrast to these intertissue/developmental patterns, the response of individual tissues to physiological stressors displayed no correlations between mRNA for PGC-1 family members and either the NRF-1 or PPAR axes. For example, in skeletal muscles, low temperature decreased PGC-1alpha transcript levels but increased mitochondrial enzyme activities (citrate synthase and cytochrome oxidase) and transcripts for COX IV and NRF-1. These results suggest that in goldfish, as in mammals, there is a regulatory relationship between (i) NRF-1 and mitochondrial gene expression and (ii) PPARs and fatty acid oxidation gene expression. In contrast to mammals, there is a divergence in the roles of the coactivators, with PGC-1alpha linked to fatty acid oxidation through PPARalpha, and PGC-1beta with a more prominent role in mediating NRF-1-dependent control of mitochondrial gene expression, as well as distinctions between their respective roles in development and physiological responsiveness. PMID- 18424680 TI - Mechanical specialization of the obliquely striated circular mantle muscle fibres of the long-finned squid Doryteuthis pealeii. AB - The centrally located, mitochondria-poor (CMP) and superficially located, mitochondria-rich (SMR) circular muscle fibres in the mantles of some squids provide one of the few known examples of specialization in an obliquely striated muscle. Little is known of the mechanical properties or of the mechanisms and performance consequences of specialization in these fibres. We combined morphological and physiological approaches to study specialization in the SMR and CMP fibres of the long-finned squid Doryteuthis pealeii. The mean thick filament length was 3.12+/-0.56 mum and 1.78+/-0.27 mum for the SMR and CMP fibres, respectively. The cross-sectional areas of the whole fibre and the core of mitochondria were significantly higher in the SMR fibres, but the area occupied by the myofilaments did not differ between the two fibre types. The area of sarcoplasmic reticulum visible in cross sections was significantly higher in CMP fibres than in SMR fibres. In live bundles of muscle fibres partially isolated from the mantle, mean peak isometric stress during tetanus was significantly greater in SMR [335 mN mm(-2) physiological cross section (pcs)] than in CMP (216 mN mm(-2) pcs) fibres. SMR fibres had a lower average twitch:tetanus ratio (SMR=0.073; CMP=0.18) and a twofold lower unloaded maximum shortening velocity at 20 degrees C (SMR=2.4 L(0) s(-1); CMP=5.1 L(0) s(-1)), where L(0) was the preparation length that yielded the highest tetanic force. The structural differences in the two muscle fibre types play a primary role in determining their mechanical properties, and the significant differences in mechanical properties indicate that squid have two muscle gears. A simple model of the mantle shows that a gradient of strain and strain rate exists across the mantle wall, with fibres adjacent to the outer edge of the mantle experiencing 1.3- to 1.4-fold lower strain and strain rate than fibres adjacent to the inner edge of the mantle. The model also predicts that the CMP fibres generate virtually no power for slow jetting while the SMR fibres are too slow to generate power for the escape jets. The transmural differences in strain and strain rate predicted by the model apply to any cylindrical animal that has circumferentially oriented muscle fibres and an internal body cavity. PMID- 18424679 TI - Reptilian uncoupling protein: functionality and expression in sub-zero temperatures. AB - Here we report the partial nucleotide sequence of a reptilian uncoupling protein (repUCP) gene from the European common lizard (Lacerta vivipara). Overlapping sequence analysis reveals that the protein shows 55%, 72% and 77% sequence homology with rat UCP1, UCP2 and UCP3, respectively, and 73% with bird and fish UCPs. RepUCP gene expression was ubiquitously detected in 4 degrees C cold acclimated lizard tissues and upregulated in muscle tissues by a 20 h exposure to sub-zero temperatures in a supercooling state or after thawing. In parallel, we show an increase in the co-activators, peroxisome proliferator-activated receptor gamma coactivator-1alpha (PGC-1alpha) and peroxisome proliferator-activated receptors (PPAR), mRNA expression, suggesting that the mechanisms regulating UCP expression may be conserved between mammals (endotherms) and reptiles (ectotherms). Furthermore, mitochondria extracted from lizard skeletal muscle showed a guanosine diphosphate (GDP)-sensitive non phosphorylating respiration. This last result indicates an inhibition of extra proton leakage mediated by an uncoupling protein, providing arguments that repUCP is functional in lizard tissues. This result is associated with a remarkable GDP-dependent increase in mitochondrial endogenous H(2)O(2) production. All together, these data support a physiological role of the repUCP in superoxide limitation by lizard mitochondria in situations of stressful oxidative reperfusion following a re-warming period in winter. PMID- 18424681 TI - Sugars are complementary resources to ethanol in foods consumed by Egyptian fruit bats. AB - Food resources are complementary for a forager if their contribution to fitness is higher when consumed together than when consumed independently, e.g. ingesting one may reduce the toxic effects of another. The concentration of potentially toxic ethanol, [EtOH], in fleshy fruit increases during ripening and affects food choices by Egyptian fruit bats, becoming deterrent at high concentrations (>/=1%). However, ethanol toxicity is apparently reduced when ingested along with some sugars; more with fructose than with sucrose or glucose. We predicted (1) that ingested ethanol is eliminated faster by bats eating fructose than by bats eating sucrose or glucose, (2) that the marginal value of fructose-containing food (food+fructose) increases with increasing [EtOH] more than the marginal value of sucrose- or glucose-containing food (food+sucrose, food+glucose), and (3) that by increasing [EtOH] the marginal value of food+sucose is incremented more than that of food+glucose. Ethanol in bat breath declined faster after they ate fructose than after eating sucrose or glucose. When food [EtOH] increased, the marginal value of food+fructose increased relative to food+glucose. However, the marginal value of food+sucrose increased with increasing [EtOH] more than food+fructose or food+glucose. Although fructose enhanced the rate at which ethanol declined in Egyptian fruit bat breath more than the other sugars, the bats treated both fructose and sucrose as complementary to ethanol. This suggests that in the wild, the amount of ethanol-containing fruit consumed or rejected by Egyptian fruit bats may be related to the fruit's own sugar content and composition, and/or the near-by availability of other sucrose- and fructose containing fruits. PMID- 18424682 TI - Functional and morphological plasticity of crocodile (Crocodylus porosus) salt glands. AB - The estuarine crocodile, Crocodylus porosus, inhabits both freshwater and hypersaline waterways and maintains ionic homeostasis by excreting excess sodium and chloride ions via lingual salt glands. In the present study, we sought to investigate the phenotypic plasticity, both morphological and functional, in the lingual salt glands of the estuarine crocodile associated with chronic exposure to freshwater (FW) and saltwater (SW) environments. Examination of haematological parameters indicated that there were no long-term disruptions to ionic homeostasis with prolonged exposure to SW. Maximal secretory rates from the salt glands of SW-acclimated animals (100.8+/-14.7 micromol 100 g(-0.7) body mass h( 1)) were almost three times greater than those of FW-acclimated animals (31.6+/ 6.2 micromol 100 g(-0.7) body mass h(-1)). There were no differences in the mass specific metabolic rate of salt gland tissue slices from FW- and SW-acclimated animals (558.9+/-49.6 and 527.3+/-142.8 microl O(2) g(-1) h(-1), respectively). Stimulation of the tissue slices from SW-acclimated animals by methacholine resulted in a 33% increase in oxygen consumption rate. There was no significant increase in the metabolic rate of tissues from FW-acclimated animals in response to methacholine. Morphologically, the secretory cells from the salt glands of SW acclimated animals were larger than those of FW-acclimated animals. In addition, there were significantly more mitochondria per unit volume in secretory tissue from SW-acclimated animals. The results from this study demonstrate that the salt glands of C. porosus are phenotypically plastic, both morphologically and functionally and acclimate to changes in environmental salinity. PMID- 18424683 TI - Duelling aphids: electrical penetration graphs reveal the value of fighting for a feeding site. AB - Horned aphids (Cerataphidini) fight each other for access to feeding sites on leaves. An attacker attempts to force another aphid to abandon its feeding site; the victor then appears to insert its stylets into the site relinquished by the loser. This study used electrical penetration graph (EPG) recordings of Astegopteryx pallida (Van der Goot) individuals to pinpoint the benefits of fighting. We show that victors take significantly less time to commence feeding in the phloem, measured from the start of probing with their stylets, compared with aphids that initiate a new or discover an abandoned site: 9.0 versus 22.9 min, respectively. We also recorded the behaviour of aphids unencumbered with the wire necessary for EPG recordings. Those adult aphids that acquired a feeding site through fighting commenced feeding on average 20 min earlier than those that did not, taking into account the time spent searching and fighting as well as probing. This study clearly establishes that horned aphids use the exact feeding site vacated by another individual and that the benefit they gain is rapid access to the phloem - more rapid, indeed, than has previously been recorded in any aphid. PMID- 18424684 TI - The influence of ontogeny and light environment on the expression of visual pigment opsins in the retina of the black bream, Acanthopagrus butcheri. AB - The correlation between ontogenetic changes in the spectral absorption characteristics of retinal photoreceptors and expression of visual pigment opsins was investigated in the black bream, Acanthopagrus butcheri. To establish whether the spectral qualities of environmental light affected the complement of visual pigments during ontogeny, comparisons were made between fishes reared in: (1) broad spectrum aquarium conditions; (2) short wavelength-reduced conditions similar to the natural environment; or (3) the natural environment (wild-caught). Microspectrophotometry was used to determine the wavelengths of spectral sensitivity of the photoreceptors at four developmental stages: larval, post settlement, juvenile and adult. The molecular sequences of the rod (Rh1) and six cone (SWS1, SWS2A and B, Rh2Aalpha and beta, and LWS) opsins were obtained and their expression levels in larval and adult stages examined using quantitative RT PCR. The changes in spectral sensitivity of the cones were related to the differing levels of opsin expression during ontogeny. During the larval stage the predominantly expressed opsin classes were SWS1, SWS2B and Rh2Aalpha, contrasting with SWS2A, Rh2Abeta and LWS in the adult. An increased proportion of long wavelength-sensitive double cones was found in fishes reared in the short wavelength-reduced conditions and in wild-caught animals, indicating that the expression of cone opsin genes is also regulated by environmental light. PMID- 18424685 TI - Acoustic pressure and particle motion thresholds in six sciaenid fishes. AB - Sciaenid fishes are important models of fish sound production, but investigations into their auditory abilities are limited to acoustic pressure measurements on five species. In this study, we used auditory brainstem response (ABR) to assess the pressure and particle acceleration thresholds of six sciaenid fishes commonly found in Chesapeake Bay, eastern USA: weakfish (Cynoscion regalis), spotted seatrout (Cynoscion nebulosus), Atlantic croaker (Micropogonias undulatus), red drum (Sciaenops ocellatus), spot (Leiostomus xanthurus) and northern kingfish (Menticirrhus saxatilis). Experimental subjects were presented with pure 10 ms tone bursts in 100 Hz steps from 100 Hz to 1.2 kHz using an airborne speaker. Sound stimuli, monitored with a hydrophone and geophone, contained both pressure and particle motion components. Sound pressure and particle acceleration thresholds varied significantly among species and between frequencies; audiograms were notably flatter for acceleration than pressure at low frequencies. Thresholds of species with diverticulae projecting anteriorly from their swim bladders (weakfish, spotted seatrout, and Atlantic croaker) were typically but not significantly lower than those of species lacking such projections (red drum, spot, northern kingfish). Sciaenids were most sensitive at low frequencies that overlap the peak frequencies of their vocalizations. Auditory thresholds of these species were used to estimate idealized propagation distances of sciaenid vocalizations in coastal and estuarine environments. PMID- 18424686 TI - Control of shell colour changes in the lobster, Panulirus cygnus. AB - The transition from juvenile to adult in the Australian western rock lobster, Panulirus cygnus (George), is preceded by a mass migration from inshore nursery reefs to offshore breeding grounds. Associated with this migration is a moult which results in the animals that are due to migrate undergoing characteristic colour change from deep red to pale pink, known as the ;white' phase, which is believed to be triggered by environmental factors. To investigate this phenomenon, the colour change of wild-caught animals was measured over two separate years in response to two important modifiers of crustacean shell colour, dietary carotenoid and background substrate colour. Changes in shell colour during this colour transition period were influenced more greatly by other factors independent of diet or background substrate and no mass colour change was induced during this time. Shell colour measurement and carotenoid quantification confirmed the presence of animals similar to wild-caught ;whites', regardless of the treatment. From these experimental observations we infer that the ;white' phase of the western rock lobster is not triggered by dietary modification or in response to background substrate. We propose that this transition is under the regulation of an ontogenetic program activated at a specific moult, which induces presently unidentified molecular changes linked to shell colour production. This unique colour transition may have evolved to provide protective camouflage during migration, and serves as an excellent model to study the genetic mechanisms underlying crustacean shell colouration. These data also provide insight into the changes in carotenoid levels induced by environmental factors, and the ability to modify crustacean shell colour in aquaculture. PMID- 18424687 TI - White matter thresholds for ischemic penumbra and infarct core in patients with acute stroke: CT perfusion study. AB - PURPOSE: To prospectively determine the parameters derived at admission computed tomographic (CT) perfusion imaging admission that best differentiate ischemic white matter that recovers from that which infarcts, with the latter retrospectively defined at a CT examination performed without contrast material (unenhanced CT) 5-7 days after the event. MATERIALS AND METHODS: Ethics committee approval and informed consent were obtained. Thirty patients with stroke underwent unenhanced CT, CT angiography, and CT perfusion studies at admission. Additionally, CT angiography was performed 24 hours after the stroke, and an unenhanced CT study was performed 5-7 days after the stroke. Five patients were excluded; the remaining patients (10 men, 15 women; mean age, 70 years +/- 13 [standard deviation]) were separated into those with recanalization (n = 16) and those without recanalization (n = 9) at 24 hours. For patients with recanalization, the final infarct was outlined on unenhanced CT images obtained 5 7 days after the event and was superimposed on coregistered maps from the CT perfusion study performed at admission. Ischemic white matter tissue (cerebral blood flow [CBF] < 14 mL/min/100 g) was identified at the admission CT perfusion study, and the penumbra was defined as the difference between the ischemic region and the infarct region. RESULTS: Infarct regions showed a matched decrease in CBF and cerebral blood volume (CBV) at admission, whereas penumbra regions showed a significant (P < .05) decrease in CBF but no change in CBV (P > .05) from contralateral values. A threshold CBF . CBV value of 8.14 was the most sensitive (95%, 20 of 21 regions) and specific (94%, 32 of 34 regions) parameter for differentiating between regions of ischemic white matter that recovered and regions of ischemic white matter that infarcted. CONCLUSION: The product of CBF and CBV derived from CT perfusion data provided the best differentiation between regions of ischemic white matter that infarcted and regions of ischemic white matter that recovered 5-7 days after a stroke. PMID- 18424688 TI - Coronary stent patency: dual-energy multidetector CT assessment in a pilot study with anthropomorphic phantom. AB - PURPOSE: To prospectively evaluate, by optimizing image acquisition and introducing alternative image postprocessing techniques, dual-energy multidetector computed tomography (CT) for depiction of the lumens of coronary artery stents placed in a moving anthropomorphic heart phantom. MATERIALS AND METHODS: Four coronary stents (2, 3, 4, and 5 mm) were examined at 64-section dual-energy multidetector CT by using a dual-detector "double-decker" imager with stacked high- and low-energy detector arrays, 0.67-mm section thickness, and 32 x 0.625-mm collimation. Simultaneous high- and low-energy data sets were acquired at 80 and 140 kVp and at 400 mAs. Cardiac motion was simulated in a moving anthropomorphic heart phantom. Stents were imaged longitudinally and axially with the phantom at rest and with it in motion. Use of an enhancement algorithm based on high- and low-energy absorption profiles was proposed. Stent lumen depiction and stent mesh delineation were quantified in terms of contrast-to-noise ratio (CNR) and kurtosis (kappa), respectively. Image quality was analyzed at univariate general linear model analysis in which peak voltage and data enhancement algorithm were dependent factors and stent orientation and cardiac motion were independent factors. RESULTS: Analysis of CNR and kappa revealed an interdependency between CNR and kappa and stent diameter: The CNR and kappa of smaller stents increased significantly when these stents were imaged at lower peak voltages in the axial plane and with the enhancement algorithm applied to the 80-kVp data sets (P < .001). The CNR and kappa of larger stents increased significantly when these stents were imaged at higher peak voltages in the longitudinal plane, and imaging of these stents benefited from the enhancement algorithm being applied to the 140-kVp data sets (P < .001). CONCLUSION: Dual energy multidetector CT performed with optimized acquisition parameters and alternative image postprocessing led to enhanced coronary stent lumen depiction to an extent beyond that achieved with single-energy multidetector CT. PMID- 18424689 TI - CT cholangiography in potential liver donors: effect of premedication with intravenous morphine on biliary caliber and visualization. AB - PURPOSE: To retrospectively determine whether premedication with intravenously administered morphine improves bile duct caliber and visualization in potential liver donors undergoing computed tomographic (CT) cholangiography. MATERIALS AND METHODS: This was a retrospective single institution study approved by the institutional review board and compliant with requirements of the HIPAA. Multidetector CT cholangiography was performed after slow infusion of 20 mL of iodipamide meglumine 52% diluted in 80 mL of normal saline in 143 consecutive potential liver donors (81 men and 62 women; mean age, 37 years); 43 received premedication with intravenous morphine sulfate (0.04 mg per kilogram of body weight) and 100 did not. Two independent readers recorded common bile duct diameter and area on axial CT images. Readers also scored bile duct visualization, including all second-order biliary branches, on a four-point scale (0, not seen; 3, excellent visualization). RESULTS: For scans obtained without and those obtained with morphine, there was no significant difference in the mean common bile duct diameter (4.1 vs 4.3 mm for reader 1 and 4.4 vs 4.6 mm for reader 2, respectively; P > .39 for both readers), in common bile duct area (20.7 vs 21.5 mm(2), for reader 1 and 21.3 vs 20.2 mm(2) for reader 2, respectively, P > .60 for both), or in second-order bile duct visualization score (2.34 vs 2.36 for reader 1 and 2.58 vs 2.50 for reader 2, respectively; P > .5 for both). CONCLUSION: The results suggest that premedication with intravenous morphine prior to CT cholangiography in potential liver donors does not increase bile duct caliber or improve biliary visualization. PMID- 18424690 TI - Comment on "Cigarette smoke-induced pulmonary inflammation is TLR4/MyD88 and IL 1R1/MyD88 signaling dependent". PMID- 18424691 TI - Tracking and trapping somatic mutations in Ig genes. PMID- 18424692 TI - Pillars article: generation of antibody diversity in the immune response of BALB/c mice to influenza virus hemagglutinin. Proc. Natl. Acad. Sci. USA, 81: 3180-3184, May 1984. PMID- 18424693 TI - IL-10: the master regulator of immunity to infection. AB - IL-10 is an anti-inflammatory cytokine. During infection it inhibits the activity of Th1 cells, NK cells, and macrophages, all of which are required for optimal pathogen clearance but also contribute to tissue damage. In consequence, IL-10 can both impede pathogen clearance and ameliorate immunopathology. Many different types of cells can produce IL-10, with the major source of IL-10 varying in different tissues or during acute or chronic stages of the same infection. The priming of these various IL-10-producing populations during infections is not well understood and it is not clear whether the cellular source of IL-10 during infection dictates its cellular target and thus its outcome. In this article we review the biology of IL-10, its cellular sources, and its role in viral, bacterial, and protozoal infections. PMID- 18424694 TI - Cutting edge: Membrane nanotubes in vivo: a feature of MHC class II+ cells in the mouse cornea. AB - Membrane nanotubes are a recently discovered form of cellular protrusion between two or more cells whose functions include cell communication, environmental sampling, and protein transfer. Although clearly demonstrated in vitro, evidence of the existence of membrane nanotubes in mammalian tissues in vivo has until now been lacking. Confocal microscopy of whole-mount corneas from wild-type, enhanced GFP chimeric mice, and Cx3cr1(gfp) transgenic mice revealed long (>300 microm) and fine (<0.8 microm diameter) membrane nanotube-like structures on bone marrow derived MHC class II(+) cells in the corneal stroma, some of which formed distinct intercellular bridges between these putative dendritic cells. The frequency of these nanotubes was significantly increased in corneas subjected to trauma and LPS, which suggests that nanotubes have an important role in vivo in cell-cell communication between widely spaced dendritic cells during inflammation. Identification of these novel cellular processes in the mammalian cornea provides the first evidence of membrane nanotubes in vivo. PMID- 18424695 TI - Cutting edge: Dendritic cells prime a high avidity CTL response independent of the level of presented antigen. AB - CTL that possess a high functional avidity are known to be optimal for the clearance of pathogens in vivo. We have shown that the amount of peptide encountered by a CD8+ CTL determines its functional avidity. Notably, in these studies nonprofessional APC were used. However, it is mature dendritic cells (DC) that are predominantly responsible for the activation of naive T cells in vivo. Whether DC also direct dose dependent-differences in avidity is unknown. In this work we examined the ability of mature DC presenting a high vs low level of peptide to generate CTL of distinct avidities. In contrast to what was observed with nonprofessional APC, CTL generated by stimulation with mature DC were of high avidity regardless of the amount of peptide presented. This DC property may promote generation of highly effective CTL that retain plasticity, which would allow the tuning of avidity in the periphery to promote optimal pathogen recognition and clearance. PMID- 18424696 TI - Cutting edge: Enhanced IL-2 signaling can convert self-specific T cell response from tolerance to autoimmunity. AB - Naive and memory T cells show differences in their response to antigenic stimulation. We examined whether this difference extended to the peripheral deletion of T cells reactive to self-Ag or, alternatively, the induction of autoimmunity. Our results show that although both populations where susceptible to deletion, memory T cells, but not naive T cells, also gave rise to autoimmunity after in vivo presentation of skin-derived self-Ags. The same migratory dendritic cells presented self-Ag to both naive and memory T cell populations, but only the latter had significant levels of the effector molecule granzyme B. Memory T cells also expressed increased levels of the high affinity IL-2 receptor chain after self-Ag recognition. Provision of IL-2 signaling using a stimulatory complex of anti-IL-2 Ab and IL-2 drove the otherwise tolerant naive T cells toward an autoimmune response. Therefore, enhanced IL-2 signaling can act as a major selector between tolerance and autoimmunity. PMID- 18424697 TI - Cutting edge: Foxp3-mediated induction of pim 2 allows human T regulatory cells to preferentially expand in rapamycin. AB - Addition of rapamycin to cultures of expanding natural CD4+CD25+Foxp3+ T regulatory cells (Tregs) helps maintain their suppressive activity, but the underlying mechanism is unclear. Pim 2 is a serine/threonine kinase that can confer rapamycin resistance. Unexpectedly, pim 2 was found to be constitutively expressed in freshly isolated, resting Tregs, but not in CD4+CD25- T effector cells. Introduction of Foxp3, but not Foxp3Delta2, into effector T cells induced pim 2 expression and conferred preferential expansion in the presence of rapamycin, indicating that Foxp3 can regulate pim 2 expression. Finally, we determined there is a positive correlation between Treg expansion and Foxp3 expression in the presence of rapamycin. Together, these results indicate that Tregs are programmed to be resistant to rapamycin, providing further rationale for why this immunosuppressive drug should be used in conjunction with expanded Tregs. PMID- 18424698 TI - Cutting edge: Overlapping functions of TLR7 and TLR9 for innate defense against a herpesvirus infection. AB - As initially demonstrated with murine cytomegalovirus (MCMV), plasmacytoid dendritic cells (pDCs) are the major source of IFN-alpha/beta in response to a variety of viruses in vivo. However, contradictory results have been obtained pertaining to the mechanisms promoting IFN-alpha/beta production by pDCs in response to MCMV. In this study we show that TLR7 and TLR9 exert redundant functions for IFN-alpha/beta, IL-12p40, and TNF-alpha production by pDCs in vivo during MCMV infection. In contrast, we confirm that systemic production of IL 12p70 strictly depends on TLR9. The combined loss of TLR7 and TLR9 recapitulates critical features of the phenotype of MyD88-deficient mice, including a dramatic decrease in systemic IFN-alpha/beta levels, an increase in viral load, and increased susceptibility to MCMV-induced mortality. This is the first demonstration of the implication of TLR7 in the recognition of a DNA virus. PMID- 18424699 TI - Lck-dependent tyrosine phosphorylation of diacylglycerol kinase alpha regulates its membrane association in T cells. AB - TCR engagement triggers phospholipase Cgamma1 activation through the Lck-ZAP70 linker of activated T cell adaptor protein pathway. This leads to generation of diacylglycerol (DAG) and mobilization of intracellular Ca(2+), both essential for TCR-dependent transcriptional responses. TCR ligation also elicits transient recruitment of DAG kinase alpha (DGKalpha) to the lymphocyte plasma membrane to phosphorylate DAG, facilitating termination of DAG-regulated signals. The precise mechanisms governing dynamic recruitment of DGKalpha to the membrane have not been fully elucidated, although Ca(2+) influx and tyrosine kinase activation were proposed to be required. We show that DGKalpha is tyrosine phosphorylated, and identify tyrosine 335 (Y335), at the hinge between the atypical C1 domains and the catalytic region, as essential for membrane localization. Generation of an Ab that recognizes phosphorylated Y335 demonstrates Lck-dependent phosphorylation of endogenous DGKalpha during TCR activation and shows that pY335DGKalpha is a minor pool located exclusively at the plasma membrane. Our results identify Y335 as a residue critical for DGKalpha function and suggest a mechanism by which Lck dependent phosphorylation and Ca(2+) elevation regulate DGKalpha membrane localization. The concerted action of these two signals results in transient, receptor-regulated DGKalpha relocalization to the site at which it exerts its function as a negative modulator of DAG-dependent signals. PMID- 18424700 TI - Virus-like display of a neo-self antigen reverses B cell anergy in a B cell receptor transgenic mouse model. AB - The ability to distinguish between self and foreign Ags is a central feature of immune recognition. For B cells, however, immune tolerance is not absolute, and factors that include Ag valency, the availability of T help, and polyclonal B cell stimuli can influence the induction of autoantibody responses. Here, we evaluated whether multivalent virus-like particle (VLP)-based immunogens could induce autoantibody responses in well-characterized transgenic (Tg) mice that express a soluble form of hen egg lysozyme (HEL) and in which B cell tolerance to HEL is maintained by anergy. Immunization with multivalent VLP-arrayed HEL, but not a trivalent form of HEL, induced high-titer Ab responses against HEL in both soluble HEL Tg mice and double Tg mice that also express a monoclonal HEL specific BCR. Induction of autoantibodies against HEL was not dependent on coadministration of strong adjuvants, such as CFA. In contrast to previous data showing the T-independent induction of Abs to foreign epitopes on VLPs, the ability of HEL-conjugated VLPs to induce anti-HEL Abs in tolerant mice was dependent on the presence of CD4(+) Th cells, and could be enhanced by the presence of pre-existing cognate T cells. In in vitro studies, VLP-conjugated HEL was more potent than trivalent HEL in up-regulating surface activation markers on purified anergic B cells. Moreover, immunization with VLP-HEL reversed B cell anergy in vivo in an adoptive transfer model. Thus, Ag multivalency and T help cooperate to reverse B cell anergy, a major mechanism of B cell tolerance. PMID- 18424701 TI - Danger signaling through the inflammasome acts as a master switch between tolerance and sensitization. AB - Efficient priming of adaptive immunity depends on danger signals provided by innate immune pathways. As an example, inflammasome-mediated activation of caspase-1 and IL-1beta is crucial for the development of reactive T cells targeting sensitizers like dinitrofluorobenzene (DNFB). Surprisingly, DNFB and dinitrothiocyanobenzene provide cross-reactive Ags yet drive opposing, sensitizing vs tolerizing, T cell responses. In this study, we show that, in mice, inflammasome-signaling levels can be modulated to turn dinitrothiocyanobenzene into a sensitizer and DNFB into a tolerizer, and that it correlates with the IL-6 and IL-12 secretion levels, affecting Th1, Th17, and regulatory T cell development. Hence, our data provide the first evidence that the inflammasome can define the type of adaptive immune response elicited by an Ag, and hint at new strategies to modulate T cell responses in vivo. PMID- 18424702 TI - Conserved mycobacterial lipoglycoproteins activate TLR2 but also require glycosylation for MHC class II-restricted T cell activation. AB - CD4(+) T cell clones derived from a leprosy lesion and patient blood were used to monitor the isolation and identification of an Ag associated with the self limited form of the disease. Biochemical purification and genetic analysis identified the T cell Ag as a conserved mycobacterial lipoglycoprotein LprG. LprG mediated activation of CD4(+) T cells required specific MHC class II restriction molecules and intracellular processing. Although LprG activated TLR2, this alone was not sufficient to stimulate or inhibit T cell activation. A striking finding was that the carbohydrate moieties of LprG were required for optimal T cell activation, because recombinant LprG produced in Escherichia coli, or recombinant LprG produced in Mycobacterium smegmatis and digested by alpha-mannosidase, did not activate T cells. This study demonstrates that the universe of bacterial T cell Ags includes lipoglycoproteins, which act as TLR2 ligands but also require glycosylation for MHC class II-restricted T cell activation in vivo. PMID- 18424703 TI - Systemic control of plasmacytoid dendritic cells by CD8+ T cells and commensal microbiota. AB - The composition of the intestinal microbial community is a distinctive individual trait that may divergently influence host biology. Because dendritic cells (DC) regulate the quality of the host response to microbiota, we evaluated DC in mice bearing distinct enteric microbial communities divergent for colitis susceptibility. Surprisingly, a selective, systemic reduction of plasmacytoid dendritic cells (pDC) was observed in isogenic mice with different microbiota: restricted flora (RF) vs specific pathogen free (SPF). This reduction was not observed in germfree mice, suggesting that the pDC deficiency was not simply due to a lack of intestinal microbial products. The microbial action was linked to cytotoxic CD8(+) T cells, since pDC in RF mice were preserved in the CD8(-/-) and perforin(-/-) genotypes, partially restored by anti-CD8beta Ab, and augmented in SPF mice bearing the TAP(-/-) genotype. Direct evidence for pDC cytolysis was obtained by rapid and selective pDC depletion in SPF mice transferred with RF CD8(+) T cells. These data indicate that commensal microbiota, via CTL activation, functionally shape systemic immune regulation that may modify risk of inflammatory disease. PMID- 18424704 TI - Pathogen proliferation governs the magnitude but compromises the function of CD8 T cells. AB - CD8+ T cell memory is critical for protection against many intracellular pathogens. However, it is not clear how pathogen virulence influences the development and function of CD8+ T cells. Salmonella typhimurium (ST) is an intracellular bacterium that causes rapid fatality in susceptible mice and chronic infection in resistant strains. We have constructed recombinant mutants of ST, expressing the same immunodominant Ag OVA, but defective in various key virulence genes. We show that the magnitude of CD8+ T cell response correlates directly to the intracellular proliferation of ST. Wild-type ST displayed efficient intracellular proliferation and induced increased numbers of OVA specific CD8+ T cells upon infection in mice. In contrast, mutants with defective Salmonella pathogenicity island II genes displayed poor intracellular proliferation and induced reduced numbers of OVA-specific CD8+ T cells. However, when functionality of the CD8+ T cell response was measured, mutants of ST induced a more functional response compared with the wild-type ST. Infection with wild-type ST, in contrast to mutants defective in pathogenicity island II genes, induced the generation of mainly effector-memory CD8+ T cells that expressed little IL-2, failed to mediate efficient cytotoxicity, and proliferated poorly in response to Ag challenge in vivo. Taken together, these results indicate that pathogens that proliferate rapidly and chronically in vivo may evoke functionally inferior memory CD8+ T cells which may promote the survival of the pathogen. PMID- 18424705 TI - Differential control of T regulatory cell proliferation and suppressive activity by mature plasmacytoid versus conventional spleen dendritic cells. AB - Anergy and suppression are cardinal features of CD4(+)CD25(+)Foxp3(+) T cells (T regulatory cells (Treg)) which have been shown to be tightly controlled by the maturation state of dendritic cells (DC). However, whether lymphoid organ DC subsets exhibit different capacities to control Treg is unclear. In this study, we have analyzed, in the rat, the role of splenic CD4(+) and CD4(-) conventional DC and plasmacytoid DC (pDC) in allogeneic Treg proliferation and suppression in vitro. As expected, in the absence of exogenous IL-2, Treg did not expand in response to immature DC. Upon TLR-induced maturation, all DC became potent stimulators of CD4(+)CD25(-) T cells, whereas only TLR7- or TLR9-matured pDC induced strong proliferation of CD4(+)CD25(+)Foxp3(+) T cells in the absence of exogenous IL-2. This capacity of pDC to reverse Treg anergy required cell contact and was partially CD86 dependent and IL-2 independent. In suppression assays, Treg strongly suppressed proliferation and IL-2 and IFN-gamma production by CD4(+)CD25(-) T cells induced by mature CD4(+) and CD4(-) DC. In contrast, upon stimulation by mature pDC, proliferating Treg suppressed IL-2 production by CD25( ) cells but not their proliferation or IFN-gamma production. Taken together, these results suggest that anergy and the suppressive function of Treg are differentially controlled by DC subsets. PMID- 18424706 TI - Tumor-induced impairment of TCR signaling results in compromised functionality of tumor-infiltrating regulatory T cells. AB - This study demonstrates, for the first time, that murine regulatory T (Treg) cells in the tumor microenvironment display both enhanced proliferation and reduced functionality. This enhanced proliferation, combined with decreased apoptosis, leads to an intratumoral accumulation of Treg cells with a unique phenotype: CD4(+)CD25(+)FoxP3(+)GITR(high)CD27(low)CD62L(-). The loss of functionality is associated with down-regulation of the TCR signaling complex, including IL-2-inducible T cell kinase. It is also demonstrated that tumor infiltrating Treg cells have impaired TCR-mediated signaling and calcium influx. Based on these findings, this study supports the hypothesis that 1) tumor infiltrating Treg cells lose functionality due to their diminished ability to become effectively activated and 2) intratumoral accumulation of Treg cells may compensate for the impaired functionality, thus maintaining immune tolerance to the tumor. PMID- 18424708 TI - CD83 expression in CD4+ T cells modulates inflammation and autoimmunity. AB - The transmembrane protein CD83 has been initially described as a maturation marker for dendritic cells. Moreover, there is increasing evidence that CD83 also regulates B cell function, thymic T cell maturation, and peripheral T cell activation. Herein, we show that CD83 expression confers immunosuppressive function to CD4(+) T cells. CD83 mRNA is differentially expressed in naturally occurring CD4(+)CD25(+) regulatory T cells, and upon activation these cells rapidly express large amounts of surface CD83. Transduction of naive CD4(+)CD25( ) T cells with CD83 encoding retroviruses induces a regulatory phenotype in vitro, which is accompanied by the induction of Foxp3. Functional analysis of CD83-transduced T cells in vivo demonstrates that these CD83(+)Foxp3(+) T cells are able to interfere with the effector phase of severe contact hypersensitivity reaction of the skin. Moreover, adoptive transfer of these cells prevents the paralysis associated with experimental autoimmune encephalomyelitis, suppresses proinflammatory cytokines IFN-gamma and IL-17, and increases antiinflammatory IL 10 in recipient mice. Taken together, our data provide the first evidence that CD83 expression can contribute to the immunosuppressive function of CD4(+) T cells in vivo. PMID- 18424707 TI - IFN-gamma and IL-17 production in experimental autoimmune encephalomyelitis depends on local APC-T cell complement production. AB - IFN-gamma- and IL-17-producing T cells autoreactive across myelin components are central to the pathogenesis of multiple sclerosis. Using direct in vivo, adoptive transfer, and in vitro systems, we show in this study that the generation of these effectors in myelin oligodendrocyte glycoprotein(35-55)-induced experimental autoimmune encephalomyelitis depends on interactions of locally produced C3a/C5a with APC and T cell C3aR/C5aR. In the absence of the cell surface C3/C5 convertase inhibitor decay-accelerating factor (DAF), but not the combined absence of DAF and C5aR and/or C3aR on APC and T cells, a heightened local autoimmune response occurs in which myelin destruction is markedly augmented in concert with markedly more IFN-gamma(+) and IL-17(+) T cell generation. The augmented T cell response is due to increased IL-12 and IL-23 elaboration by APCs together with increased T cell expression of the receptors for each cytokine. The results apply to initial generation of the IL-17 phenotype because naive CD62L(high) Daf1(-/-) T cells produce 3-fold more IL-17 in response to TGF-beta and IL-6, whereas CD62L(high) Daf1(-/-)C5aR(-/-)C3aR(-/-) T cells produce 4-fold less. PMID- 18424709 TI - Deacetylase activity is required for STAT5-dependent GM-CSF functional activity in macrophages and differentiation to dendritic cells. AB - After interaction with its receptor, GM-CSF induces phosphorylation of the beta chain in two distinct domains in macrophages. One induces activation of mitogen activated protein kinases and the PI3K/Akt pathway, and the other induces JAK2 STAT5. In this study we describe how trichostatin A (TSA), which inhibits deacetylase activity, blocks JAK2-STAT5-dependent gene expression but not the expression of genes that depend on the signal transduction induced by the other domain of the receptor. TSA treatment inhibited the GM-CSF-dependent proliferation of macrophages by interfering with c-myc and cyclin D1 expression. However, M-CSF-dependent proliferation, which requires ERK1/2, was unaffected. Protection from apoptosis, which involves Akt phosphorylation and p21(waf-1) expression, was not modified by TSA. GM-CSF-dependent expression of MHC class II molecules was inhibited because CIITA was not induced. The generation of dendritic cells was also impaired by TSA treatment because of the inhibition of IRF4, IRF2, and RelB expression. TSA mediates its effects by preventing the recruitment of RNA polymerase II to the promoter of STAT5 target genes and by inhibiting their expression. However, this drug did not affect STAT5A or STAT5B phosphorylation or DNA binding. These results in GM-CSF-treated macrophages reveal a relationship between histone deacetylase complexes and STAT5 in the regulation of gene expression. PMID- 18424710 TI - HIV-1-specific T cell precursors with high proliferative capacity correlate with low viremia and high CD4 counts in untreated individuals. AB - Evidences have recently suggested that the preservation of vaccine-induced memory rather than effector T cells is essential for better outcome and survival following pathogenic SIV challenge in macaques. However, an equivalent demonstration in humans is missing, and the immune correlates of HIV-1 control have been only partially characterized. We focused on the quantification of Ag specific T cell precursors with high proliferative capacity (PHPC) using a peptide-based cultured IFN-gamma ELISPOT assay (PHPC assay), which has been shown to identify expandable memory T cells. To determine which responses correlate with viral suppression and positive immunologic outcome, PBMC from 32 chronically untreated HIV-1-infected individuals were evaluated in response to peptide pools, representing the complete HIV-1 Gag, Nef, and Rev proteins, by PHPC and IFN-gamma ELISPOT assay, which instead identifies effector T cells with low proliferative capacity. High magnitude of Gag-specific PHPC, but not ELISPOT, responses significantly correlated with low plasma viremia, due to responses directed toward p17 and p15 subunits. Only Gag p17-specific PHPC response significantly correlated with high CD4 counts. Analysis of 20 additional PBMC samples from an independent cohort of chronically untreated HIV-1-infected individuals confirmed the correlation between Gag p17-specific PHPC response and either plasma viremia (inverse correlation) or CD4 counts (direct correlation). Our results indicate that the PHPC assay is quantitatively and qualitatively different from the ELISPOT assay, supporting that different T cell populations are being evaluated. The PHPC assay might be an attractive option for individual patient management and for the design and testing of therapeutic and prophylactic vaccines. PMID- 18424711 TI - Regulatory T cells inhibit dendritic cells by lymphocyte activation gene-3 engagement of MHC class II. AB - Lymphocyte activation gene-3 (LAG-3) is a CD4-related transmembrane protein expressed by regulatory T cells that binds MHC II on APCs. It is shown in this study that during Treg:DC interactions, LAG-3 engagement with MHC class II inhibits DC activation. MHC II cross-linking by agonistic Abs induces an ITAM mediated inhibitory signaling pathway, involving FcgammaRgamma and ERK-mediated recruitment of SHP-1 that suppresses dendritic cell maturation and immunostimulatory capacity. These data reveal a novel ITAM-mediated inhibitory signaling pathway in DCs triggered by MHC II engagement of LAG-3, providing a molecular mechanism in which regulatory T cells may suppress via modulating DC function. PMID- 18424713 TI - IL-12 signaling drives CD8+ T cell IFN-gamma production and differentiation of KLRG1+ effector subpopulations during Toxoplasma gondii Infection. AB - IFN-gamma-producing CD8(+) T lymphocytes are essential effector cells that mediate protective immunity during murine toxoplasmosis, and yet their effector development remains poorly characterized. Vaccination with the carbamoyl phosphate synthase (CPS) mutant strain of Toxoplasma gondii was used to examine the CD8(+) T cell response in the peritoneal effector site. Four CTL subpopulations with varying effector potentials were defined based on the expression of effector molecules and the cell surface activation markers CD62L and killer cell lectin-like receptor G1 (KLRG1). Further phenotypic analysis revealed that the acquisition of KLRG1 among effector subpopulations correlated with the down-regulation of both IL-7R and CD27, suggesting that KLRG1 marks dominant, end-stage effector cells. Using gene-targeted mice, we tested the in vivo requirements of key IL-12 signaling components for effector CTL differentiation. Contrary to established models of viral and bacterial infection, CD8(+) T cell-intrinsic IL-12 signaling was required for the generation of IFN gamma-producing CTLs in response to T. gondii. Importantly, the development of the KLRG1(+) effector subpopulations, but not the memory precursor-containing KLRG1(-) effector subset, was critically reliant on IL-12. Furthermore, IL-12 signaling-dependent T-bet expression was also found to be important for differentiation of KLRG1(+) effectors. Our results underscore a vital role for IL 12 in not only the induction of IFN-gamma expression but also in the development of heterogeneous subpopulations of effector CD8(+) T cells generated in response to the intracellular parasite T. gondii. PMID- 18424714 TI - An interaction between CD200 and monoclonal antibody agonists to CD200R2 in development of dendritic cells that preferentially induce populations of CD4+CD25+ T regulatory cells. AB - In previous studies we reported that while interaction between the relatively ubiquitously expressed molecule CD200 and one of its receptors, CD200R1, resulted in direct suppression of alloreactivity, engagement of alternate receptors led instead to altered differentiation of dendritic cells (DCs) from marrow precursors, which could in turn foster development of Foxp3(+) regulatory T cells. We have explored this effect of engagement of alternate receptors by using a monoclonal agonist Ab to CD200R2 and investigating expression of TLRs on DCs induced in vivo and in vitro after CD200 stimulation in mice in which the gene encoding CD200R1 was deleted. CD200 stimulation was achieved by using either a soluble form of CD200 (CD200Fc) or overexpression of CD200 as a doxycycline inducible transgene. Although broadly similar effects were seen, consistent with the hypothesis that triggering of CD200R2 does produce DCs with a characteristic TLR repertoire, there are subtle differences in suppression of alloreactivity achieved by CD200 delivered in these two manners, which is consistent with a complexity of CD200:CD200R engagement not previously appreciated. PMID- 18424712 TI - Disruption of mutually negative regulatory feedback loop between interferon inducible p202 protein and the E2F family of transcription factors in lupus-prone mice. AB - Studies have identified IFN-inducible Ifi202 gene as a lupus susceptibility gene (encoding p202 protein) in mouse models of lupus disease. However, signaling pathways that regulate the Ifi202 expression in cells remain to be elucidated. We found that steady-state levels of Ifi202 mRNA and protein were high in mouse embryonic fibroblasts (MEFs) from E2F1 knockout (E2F1(-/-)) and E2F1 and E2F2 double knockout (E2F1(-/-)E2F2(-/-)) mice than isogenic wild-type MEFs. Moreover, overexpression of E2F1 in mouse fibroblasts decreased expression of p202. Furthermore, expression of E2F1, but not E2F4, transcription factor in mouse fibroblasts repressed the activity of 202-luc-reporter in promoter-reporter assays. Interestingly, the E2F1-mediated transcriptional repression of the 202 luc-reporter was independent of p53 and pRb expression. However, the repression was dependent on the ability of E2F1 to bind DNA. We have identified a potential E2F DNA-binding site in the 5'-regulatory region of the Ifi202 gene, and mutations in this E2F DNA-binding site reduced the E2F1-mediated transcriptional repression of 202-luc-reporter. Because p202 inhibits the E2F1-mediated transcriptional activation of genes, we compared the expression of E2F1 and its target genes in splenic cells from lupus-prone B6.Nba2 congenic mice, which express increased levels of p202, with age-matched C57BL/6 mice. We found that increased expression of Ifi202 in the congenic mice was associated with inhibition of E2F1-mediated transcription and decreased expression of E2F1 and its target genes that encode proapoptotic proteins. Our observations support the idea that increased Ifi202 expression in certain strains of mice contributes to lupus susceptibility in part by inhibiting E2F1-mediated functions. PMID- 18424715 TI - Conversion of tyrosine to the inflammation-associated analog 3'-nitrotyrosine at either TCR- or MHC-contact positions can profoundly affect recognition of the MHC class I-restricted epitope of lymphocytic choriomeningitis virus glycoprotein 33 by CD8 T cells. AB - Immunohistochemical detection of increased levels of protein-associated nitrotyrosine has become widely used as a surrogate marker of in situ inflammation. However, the potential consequences of protein-associated nitrotyrosine formation in terms of cellular immune recognition has received surprisingly little attention. Using a well-defined I-E(K)-restricted epitope of pigeon cytochrome c, we previously demonstrated that conversion of a single tyrosine residue to nitrotyrosine can have a profound effect on recognition by CD4 T cells. In this study, we used the MHC class I-restricted epitope of lymphocytic choriomeningitis virus glycoprotein (gp33) to demonstrate that conversion of tyrosine to nitrotyrosine can also profoundly affect recognition of MHC class I-restricted epitopes. Conversion of the Y4 residue of the gp33 epitope to nitrotyrosine completely abrogated recognition by gp33-specific T cells from P14 TCR-transgenic mice. In contrast, CD8(+) T cells specific for "nitrated gp33" (NY-gp33) can be readily elicited in C57BL/6 mice after immunization with NY-gp33 peptide. Interestingly, T-T hybridomas specific for NY-gp33 peptide were found to fall into two distinct subsets, being specific for NY-gp33 presented in the context of either H-2D(b) or H-2K(b). This latter result is surprising in light of previous structural studies showing that Y4 comprises a critical TCR-contact residue when presented by H-2D(b) but that the same residue points downward into the peptide-binding groove of the MHC when presented by H-2K(b). Together, these results indicate that nitrotyrosine formation can impact T cell recognition both directly, through alteration of TCR-contact residues, or indirectly, through alterations in MHC-contact positions. PMID- 18424716 TI - Plasmacytoid dendritic cells migrate in afferent skin lymph. AB - Conventional dendritic cells enter lymph nodes by migrating from peripheral tissues via the lymphatic route, whereas plasmacytoid dendritic cells (pDC), also called IFN-producing cells (IPC), are described to gain nodes from blood via the high endothelial venules. We demonstrate here that IPC/pDC migrate in the afferent lymph of two large mammals. In sheep, injection of type A CpG oligodinucleotide (ODN) induced lymph cells to produce type I IFN. Furthermore, low-density lymph cells collected at steady state produced type I IFN after stimulation with type A CpG ODN and enveloped viruses. Sheep lymph IPC were found within a minor B(neg)CD11c(neg) subset expressing CD45RB. They presented a plasmacytoid morphology, expressed high levels of TLR-7, TLR-9, and IFN regulatory factor 7 mRNA, induced IFN-gamma production in allogeneic CD4(pos) T cells, and differentiated into dendritic cell-like cells under viral stimulation, thus fulfilling criteria of bona fide pDC. In mini-pig, a CD4(pos)SIRP(pos) subset in afferent lymph cells, corresponding to pDC homologs, produced type I IFN after type A CpG-ODN triggering. Thus, pDC can link innate and acquired immunity by migrating from tissue to draining node via lymph, similarly to conventional dendritic cells. PMID- 18424717 TI - Preferential development of CD4 and CD8 T regulatory cells in RasGRP1-deficient mice. AB - RasGRP1 and Sos are two Ras-guanyl-nucleotide exchange factors that link TCR signal transduction to Ras and MAPK activation. Recent studies demonstrate positive selection of developing thymocytes is crucially dependent on RasGRP1, whereas negative selection of autoreactive thymocytes appears to be RasGRP1 independent. However, the role of RasGRP1 in T regulatory (Treg) cell development and function is unknown. In this study, we characterized the development and function of CD4(+)CD25(+)Foxp3(+) and CD8(+)CD44(high)CD122(+) Treg lineages in RasGRP1(-/-) mice. Despite impaired CD4 Treg cell development in the thymus, the periphery of RasGRP1(-/-) mice contained significantly increased frequencies of CD4(+)Foxp3(+) Treg cells that possessed a more activated cell surface phenotype. Furthermore, on a per cell basis, CD4(+)Foxp3(+) Treg cells from mutant mice are more suppressive than their wild-type counterparts. Our data also suggest that the lymphopenic environment in the mutant mice plays a dominant role of favored peripheral development of CD4 Treg cells. These studies suggest that whereas RasGRP1 is crucial for the intrathymic development of CD4 Treg cells, it is not required for their peripheral expansion and function. By contrast to CD4(+)CD25(+)Foxp3(+) T cells, intrathymic development of CD8(+)CD44(high)CD122(+) Treg cells is unaffected by the RasGRP1(-/-) mutation. Moreover, RasGRP1(-/-) mice contained greater numbers of CD8(+)CD44(high)CD122(+) T cells in the spleen, relative to wild-type mice. Activated CD8 Treg cells from RasGRP1(-/-) mice retained their ability to synthesize IL-10 and suppress the proliferation of wild-type CD8(+)CD122(-) T cells, albeit at a much lower efficiency than wild-type CD8 Treg cells. PMID- 18424718 TI - SUMOylation regulates the transcriptional activity of JunB in T lymphocytes. AB - The AP-1 family member JunB is a critical regulator of T cell function. JunB is a transcriptional activator of various cytokine genes, such as IL-2, IL-4, and IL 10; however, the post-translational modifications that regulate JunB activity in T cells are poorly characterized. We show here that JunB is conjugated with small ubiquitin-like modifier (SUMO) on lysine 237 in resting and activated primary T cells and T cell lines. Sumoylated JunB associated with the chromatin-containing insoluble fraction of cells, whereas nonsumoylated JunB was also in the soluble fraction. Blocking JunB sumoylation by mutation or use of a dominant-negative form of the SUMO-E2 Ubc-9 diminished its ability to transactivate IL-2 and IL-4 reporter genes. In contrast, nonsumoylable JunB mutants showed unimpaired activity with reporter genes controlled by either synthetic 12-O tetradecanoylphorbol-13-acetate response elements or NF-AT/AP-1 and CD28RE sites derived from the IL-2 promoter. Ectopic expression of JunB in activated human primary CD4(+) T cells induced activation of the endogenous IL-2 promoter, whereas the nonsumoylable JunB mutant did not. Thus, our work demonstrates that sumoylation of JunB regulates its ability to induce cytokine gene transcription and likely plays a critical role in T cell activation. PMID- 18424719 TI - Prevention of allograft tolerance by bacterial infection with Listeria monocytogenes. AB - Exposure to certain viruses and parasites has been shown to prevent the induction of transplantation tolerance in mice via the generation of cross-reactive memory T cell responses or the induction of bystander activation. Bacterial infections are common in the perioperative period of solid organ allograft recipients in the clinic, and correlations between bacterial infections and acute allograft rejection have been reported. However, whether bacterial infections at the time of transplantation have any effect on the generation of transplantation tolerance remains to be established. We used the Gram-positive intracellular bacterium Listeria monocytogenes (LM) as a model pathogen because its effects on immune responses are well described. Perioperative LM infection prevented cardiac and skin allograft acceptance induced by anti-CD154 and donor-specific transfusion in mice. LM-mediated rejection was not due to the generation of cross-reactive T cells and was largely independent of signaling via MyD88, an adaptor for most TLRs, IL-1, and IL-18. Instead, transplant rejection following LM infection was dependent on the expression of the phagosome-lysing pore former listeriolysin O and on type I IFN receptor signaling. Our results indicate that bacterial exposure at the time of transplantation can antagonize tolerogenic regimens by enhancing alloantigen-specific immune responses independently of the generation of cross-reactive memory T cells. PMID- 18424720 TI - Fungal proteases induce Th2 polarization through limited dendritic cell maturation and reduced production of IL-12. AB - Allergens are capable of polarizing the T cell immune response toward a Th2 cytokine profile in a process that is mediated by dendritic cells (DCs). Proteases derived from Aspergillus species (Aspergillus proteases; AP) have been shown to induce a Th2-like immune response when administered directly to the airway and without adjuvant or prior priming immunizations at sites remote from the lung in models of allergic airway disease. To explore mechanisms that underlie the Th2 immune response, we have investigated the effect of AP on DC function. We found that human DCs derived from CD14(+) monocytes from healthy donors underwent partial maturation when incubated with AP. Naive allogeneic T cells primed with AP-activated DCs proliferated and displayed enhanced production of IL-4 and reduced expression of IFN-gamma as compared with naive T cells primed with LPS-activated DCs. Global gene expression analysis of DCs revealed relatively low expression of IL-12p40 in AP-activated DCs as compared with those activated by LPS, and this was confirmed at the protein level by ELISA. Exogenous IL-12p70 added to cocultures of DCs and T cells resulted in reduced IL-4 and increased IFN-gamma expression when DCs were activated with AP. When the proteolytic activity of AP was neutralized by chemical inactivation it failed to up-regulate costimulatory molecules on DCs, and these DCs did not prime a Th2 response in naive T cells. These findings provide a mechanism for explaining how proteolytically active allergens could preferentially induce Th2 responses through limited maturation of DCs with reduced production of IL-12. PMID- 18424721 TI - Inorganic arsenic activates reduced NADPH oxidase in human primary macrophages through a Rho kinase/p38 kinase pathway. AB - Inorganic arsenic is an immunotoxic environmental contaminant to which millions of humans are chronically exposed. We recently demonstrated that human primary macrophages constituted a critical target for arsenic trioxide (As(2)O(3)), an inorganic trivalent form. To specify the effects of arsenic on macrophage phenotype, we investigated in the present study whether As(2)O(3) could regulate the activity of NADPH oxidase, a major superoxide-generating enzymatic system in human phagocytes. Our results show that superoxide levels were significantly increased in a time-dependent manner in blood monocyte-derived macrophages treated with 1 muM As(2)O(3) for 72 h. Concomitantly, As(2)O(3) induced phosphorylation and membrane translocation of the NADPH oxidase subunit p47(phox) and it also increased translocation of Rac1 and p67(phox). Apocynin, a selective inhibitor of NADPH oxidases, prevented both p47(phox) translocation and superoxide production. NADPH oxidase activation was preceded by phosphorylation of p38-kinase in As(2)O(3)-treated macrophages. The p38-kinase inhibitor SB 203580 prevented phosphorylation and translocation of p47(phox) and subsequent superoxide production. Pretreatment of macrophages with the Rho-kinase inhibitor Y-27632 was found to mimic inhibitory effects of SB-203580 and to prevent As(2)O(3)-induced phosphorylation of p38 kinase. Treatment with As(2)O(3) also resulted in an increased secretion of the proinflammatory chemokine CCL18 that was fully inhibited by both apocynin and SB-203580. Taken together, our results demonstrate that As(2)O(3) induced a marked activation of NADPH oxidase in human macrophages, likely through stimulation of a Rho-kinase/p38-kinase pathway, and which may contribute to some of the deleterious effects of inorganic arsenic on macrophage phenotype. PMID- 18424722 TI - Reovirus activates human dendritic cells to promote innate antitumor immunity. AB - Oncolytic viruses can exert their antitumor activity via direct oncolysis or activation of antitumor immunity. Although reovirus is currently under clinical investigation for the treatment of localized or disseminated cancer, any potential immune contribution to its efficacy has not been addressed. This is the first study to investigate the ability of reovirus to activate human dendritic cells (DC), key regulators of both innate and adaptive immune responses. Reovirus induced DC maturation and stimulated the production of the proinflammatory cytokines IFN-alpha, TNF-alpha, IL-12p70, and IL-6. Activation of DC by reovirus was not dependent on viral replication, while cytokine production (but not phenotypic maturation) was inhibited by blockade of PKR and NF-kappaB signaling. Upon coculture with autologous NK cells, reovirus-activated DC up-regulated IFN gamma production and increased NK cytolytic activity. Moreover, short-term coculture of reovirus-activated DC with autologous T cells also enhanced T cell cytokine secretion (IL-2 and IFN-gamma) and induced non-Ag restricted tumor cell killing. These data demonstrate for the first time that reovirus directly activates human DC and that reovirus-activated DC stimulate innate killing by not only NK cells, but also T cells, suggesting a novel potential role for T cells in oncolytic virus-induced local tumor cell death. Hence reovirus recognition by DC may trigger innate effector mechanisms to complement the virus's direct cytotoxicity, potentially enhancing the efficacy of reovirus as a therapeutic agent. PMID- 18424723 TI - Phospholipase D1 plays a key role in TNF-alpha signaling. AB - The primary characteristic features of any inflammatory or infectious lesions are immune cell infiltration, cellular proliferation, and the generation of proinflammatory mediators. TNF-alpha is a potent proinflammatory and immuno regulatory cytokine. Decades of research have been focused on the physiological/pathophysiological events triggered by TNF-alpha. However, the signaling network initiated by TNF-alpha in human leukocytes is still poorly understood. In this study, we report that TNF-alpha activates phospholipase D1 (PLD1), in a dose-dependent manner, and PLD1 is required for the activation of sphingosine kinase and cytosolic calcium signals. PLD1 is also required for NFkappaB and ERK1/2 activation in human monocytic cells. Using antisense oligonucleotides to reduce specifically the expression of PLD isozymes showed PLD1, but not PLD2, to be coupled to TNF-alpha signaling and that PLD1 is required to mediate receptor activation of sphingosine kinase and calcium transients. In addition, the coupling of TNF-alpha to activation of the phosphorylation of ERK1/2 and the activation of NFkappaB were inhibited by pretreating cells with antisense to PLD1, but not to PLD2; thus, demonstrating a specific requirement for PLD1. Furthermore, use of antisense oligonucleotides to reduce expression of PLD1 or PLD2 demonstrated that PLD1 is required for TNF alpha-induced production of several important cytokines, such as IL-1beta, IL-5, IL-6, and IL-13, in human monocytes. These studies demonstrate the critical role of PLD1 in the intracellular signaling cascades initiated by TNF-alpha and its functional role for coordinating the signals to inflammatory responses. PMID- 18424725 TI - Protective immunosurveillance and therapeutic antitumor activity of gammadelta T cells demonstrated in a mouse model of prostate cancer. AB - In contrast to Ag-specific alphabeta T cells, gammadelta T cells can kill malignantly transformed cells in a manner that does not require the recognition of tumor-specific Ags. Although such observations have contributed to the emerging view that gammadelta T cells provide protective innate immunosurveillance against certain malignancies, particularly those of epithelial origin, they also provide a rationale for developing novel clinical approaches to exploit the innate antitumor properties of gammadelta T cells for the treatment of cancer. Using TRAMP, a transgenic mouse model of prostate cancer, proof-of concept studies were performed to first establish that gammadelta T cells can indeed provide protective immunosurveillance against spontaneously arising mouse prostate cancer. TRAMP mice, which predictably develop prostate adenocarcinoma, were backcrossed with gammadelta T cell-deficient mice (TCRdelta(-/-) mice) yielding TRAMP x TCRdelta(-/-) mice, a proportion of which developed more extensive disease compared with control TRAMP mice. By extension, these findings were then used as a rationale for developing an adoptive immunotherapy model for treating prostate cancer. Using TRAMP-C2 cells derived from TRAMP mice (C57BL/6 genetic background), disease was first established in otherwise healthy wild-type C57BL/6 mice. In models of localized and disseminated disease, tumor-bearing mice treated i.v. with supraphysiological numbers of syngeneic gammadelta T cells (C57BL/6-derived) developed measurably less disease compared with untreated mice. Disease-bearing mice treated i.v. with gammadelta T cells also displayed superior survival compared with untreated mice. These findings provide a biological rationale for clinical trials designed to adoptively transfer ex vivo expanded autologous gammadelta T cells for the treatment of prostate cancer. PMID- 18424724 TI - TLR6 modulates first trimester trophoblast responses to peptidoglycan. AB - Intrauterine bacterial infections are a well-established cause of pregnancy complications. One key observation in a number of abnormal pregnancies is that placental apoptosis is significantly elevated. First trimester trophoblast cells are known to express TLR1 and TLR2 and to undergo apoptosis following exposure to Gram-positive bacterial peptidoglycan (PDG). Thus, the objectives of this study were to determine whether PDG-induced pregnancy complications are associated with placental apoptosis and to characterize the cellular mechanisms involved. We have demonstrated, using an animal model, that delivery of PDG to pregnant mice early in gestation resulted in highly elevated placental apoptosis, evidenced by trophoblast M-30 and active caspase 3 immunostaining. Using an in vitro model of human first trimester trophoblasts, apoptosis induced by PDG was found to be mediated by both TLR1 and TLR2 and that this could be blocked by the presence of TLR6. Furthermore, in the presence of TLR6, exposure to PDG resulted in trophoblast NF-kappaB activation and triggered these cells to secrete IL-8 and IL 6. The findings of this study suggest that a Gram-positive bacterial infection, through TLR2 and TLR1, may directly promote the elevated trophoblast cell death and that this may be the underlying mechanism of pregnancy complications, such as preterm delivery. Furthermore, the expression of TLR6 may be a key factor in determining whether the response to PDG would be apoptosis or inflammation. PMID- 18424726 TI - Characterization of IL-10-secreting T cells derived from regulatory CD4+CD25+ cells by the TIRC7 surface marker. AB - Natural CD25(+)CD4(+) regulatory T cells (Treg) are essential for self-tolerance and for the control of T cell-mediated immune pathologies. However, the identification of Tregs in an ongoing immune response or in inflamed tissues remains elusive. Our experiments indicate that TIRC7, T cell immune response cDNA 7, a novel membrane molecule involved in the regulation of T lymphocyte activation, identifies two Treg subsets (CD25(low)TIRC7(+) and CD25(high)TIRC7( )) that are characterized by the expression of Foxp3 and a suppressive activity in vitro and in vivo. We also showed that the CD25(low)TIRC7(+) subset represents IL-10-secreting Tregs in steady state, which is accumulated intratumorally in a tumor-bearing mice model. Blockade of the effect of IL-10 reversed the suppression imposed by the CD25(low)TIRC7(+) subset. Interestingly, these IL-10 secreting cells derived from the CD25(high)TIRC7(-) subset, both in vitro and in vivo, in response to tumoral Ags. Our present results strongly support the notion that, in the pool of natural Tregs, some cells can recognize foreign Ags and that this recognition is an essential step in their expansion and suppressive activity in vivo. PMID- 18424727 TI - Suppression of normal and malignant kit signaling by a bispecific antibody linking kit with CD300a. AB - Through its receptor Kit (CD117), stem cell factor (SCF) critically regulates human mast cell (MC) differentiation, survival, priming, and activation. The dominance of SCF in setting these parameters compels stringent contra-regulation to maintain a balanced MC phenotype. We have synthesized a library of bispecific Ab fragments to examine the effect of linking Kit with CD300a. In this study, we report that CD300a exerts a strong inhibitory effect on Kit-mediated SCF-induced signaling, consequently impairing MC differentiation, survival, and activation in vitro. This effect derives from Kit-mediated tyrosine phosphorylation of CD300a and recruitment of the SHIP-1 but not of SH2-containing protein phosphatase 1. CD300a inhibits the constitutive activation of the human leukemic HMC-1 cells but not their survival. Finally, CD300a abrogates the allergic reaction induced by SCF in a murine model of cutaneous anaphylaxis. Our findings highlight CD300a as a novel regulator of Kit in human MC and suggest roles for this receptor as a suppressor of Kit signaling in MC-related disorders. PMID- 18424728 TI - Loss of STAT3 in CD4+ T cells prevents development of experimental autoimmune diseases. AB - Th17 cells are implicated in CNS autoimmune diseases. We show that mice with targeted-deletion of Stat3 in CD4(+) T cells (CD4(Stat3)(-/-)) do not develop experimental autoimmune uveoretinitis (EAU) or experimental autoimmune encephalomyelitis. Defective Th17 differentiation noted in CD4(Stat3)(-/-) mice is compensated by exaggerated increases in Foxp3-, IL-10-, IL-4-, and IFN-gamma expressing T cells, suggesting critical roles of STAT3 in shaping Ag-specific CD4(+) T cell repertoire. In mice with EAU, a high percentage of IL-17-expressing T cells in their peripheral lymphoid organs also secrete IFN-gamma while these double-expressors are absent in CD4(Stat3)(-/-) and wild-type mice without EAU, raising the intriguing possibility that uveitis maybe mediated by Th17 and IL-17 expressing Th1 cells. Resistance of Stat3-deficient mice to EAU derives in part from an inability of uveitogenic Th17 and Th1 cells to enter eyes or brain of the CD4(Stat3)(-/-) mouse because of the reduction in the expression of activated alpha4/beta1 integrins on CD4(Stat3)(-/-) T cells. Adoptive transfer of activated interphotoreceptor retinoid-binding protein-specific uveitogenic T cells induced in CD4(Stat3)(-/-) mice a severe EAU characterized by development of retinal folds, infiltration of inflammatory cells into the retina, and destruction of retinal architecture, underscoring our contention that the loss of STAT3 in CD4(+) T cells results in an intrinsic developmental defect that renders CD4(Stat3)(-/-) resistant to CNS inflammatory diseases. STAT3 requirement for IL 17 production by Th17, generation of double positive T cells expressing IL-17 and IFN-gamma, and for T cell trafficking into CNS tissues suggests that STAT3 may be a therapeutic target for modulating uveitis, sceritis, or multiple sclerosis. PMID- 18424729 TI - Long-term control of alloreactive B cell responses by the suppression of T cell help. AB - Alloantibodies can play a key role in acute and chronic allograft rejection. However, relatively little is known of factors that control B cell responses following allograft tolerance induction. Using 3-83 Igi mice expressing an alloreactive BCR, we recently reported that allograft tolerance was associated with the sustained deletion of the alloreactive B cells at the mature, but not the immature, stage. We have now investigated the basis for the long-term control of alloreactive B cell responses in a non-BCR-transgenic model of C57BL/6 cardiac transplantation into BALB/c recipients treated with anti-CD154 and transfusion of donor-specific spleen cells. We demonstrate that the long-term production of alloreactive Abs by alloreactive B cells is actively regulated in tolerant BALB/c mice through the dominant suppression of T cell help. Deletion of CD25(+) cells resulted in a loss of tolerance and an acquisition of the ability to acutely reject allografts. In contrast, the restoration of alloantibody responses required both the deletion of CD25(+) cells and the reconstitution of alloreactive B cells. Collectively, these data suggest that alloreactive B cell responses in this model of tolerance are controlled by dominant suppression of T cell help as well as the deletion of alloreactive B cells in the periphery. PMID- 18424730 TI - Carcinoembryonic antigen-related cell adhesion molecule 1 inhibits proximal TCR signaling by targeting ZAP-70. AB - The long cytoplasmic tail (CT) isoforms of carcinoembryonic Ag-related cell adhesion molecule 1 (CEACAM1) are expressed on activated human T cells and possess two ITIM motifs in the CT. These isoforms of CEACAM1 are inhibitory for T cell responses initiated by the TCR/CD3 complex with the inhibition dependent upon the ITIMs of CEACAM1 and Src homology 2 domain-containing phosphatase 1 (SHP 1). However, the mechanism by which this inhibition occurs in T cells is unknown. We demonstrate here that the Src family kinase, Lck, and the ability of CEACAM1 to bind homophilically are required for the ITIM phosphorylation of CEACAM1 that is a prerequisite for CEACAM1 association with SHP-1. We further show that CEACAM1 associates with and recruits SHP-1 to the TCR/CD3 complex leading to decreased phosphorylation of CD3-zeta and ZAP-70 and consequently decreased activation of the elements downstream of ZAP-70. This is physiologically relevant because extinction of SHP-1 expression or blockade of homophilic binding by CEACAM1 using a Fab that specifically recognizes the homophilic binding region of human CEACAM1 increases the cytolytic function initiated by the TCR/CD3 complex. These studies show that long CT isoforms of CEACAM1 orchestrate an inhibitory program that abrogates extremely proximal events downstream of the TCR/CD3 complex by focusing on the activation of ZAP-70. PMID- 18424731 TI - Antigen receptor editing in anti-DNA transitional B cells deficient for surface IgM. AB - In response to encounter with self-Ag, autoreactive B cells may undergo secondary L chain gene rearrangement (receptor editing) and change the specificity of their Ag receptor. Knowing at what differentiative stage(s) developing B cells undergo receptor editing is important for understanding how self-reactive B cells are regulated. In this study, in mice with Ig transgenes coding for anti-self (DNA) Ab, we report dsDNA breaks indicative of ongoing secondary L chain rearrangement not only in bone marrow cells with a pre-B/B cell phenotype but also in immature/transitional splenic B cells with little or no surface IgM (sIgM( /low)). L chain-edited transgenic B cells were detectable in spleen but not bone marrow and were still found to produce Ab specific for DNA (and apoptotic cells), albeit with lower affinity for DNA than the unedited transgenic Ab. We conclude that L chain editing in anti-DNA-transgenic B cells is not only ongoing in bone marrow but also in spleen. Indeed, transfer of sIgM(-/low) anti-DNA splenic B cells into SCID mice resulted in the appearance of a L chain editor (Vlambdax) in the serum of engrafted recipients. Finally, we also report evidence for ongoing L chain editing in sIgM(low) transitional splenic B cells of wild-type mice. PMID- 18424732 TI - Functional activity of MD-2 polymorphic variant is significantly different in soluble and TLR4-bound forms: decreased endotoxin binding by G56R MD-2 and its rescue by TLR4 ectodomain. AB - MD-2 is an essential component of endotoxin (LPS) sensing, binding LPS independently and when bound to the ectodomain of the membrane receptor TLR4. Natural variation of proteins involved in the LPS-recognition cascade such as the LPS-binding protein, CD14, and TLR4, as well as proteins involved in intracellular signaling downstream of LPS binding, affect the cellular response to endotoxin and host defense against bacterial infections. We now describe the functional properties of two nonsynonymous coding polymorphisms of MD-2, G56R and P157S, documented in HapMap. As predicted from the MD-2 structure, the P157S mutation had little or no effect on MD-2 function. In contrast, the G56R mutation, located close to the LPS-binding pocket, significantly decreased cellular responsiveness to LPS. Soluble G56R MD-2 showed markedly reduced LPS binding that was to a large degree rescued by TLR4 coexpression or presence of TLR4 ectodomain. Thus, cells that express TLR4 without MD-2 and whose response to LPS depends on ectopically produced MD-2 were most affected by expression of the G56R variant of MD-2. Coexpression of wild-type and G56R MD-2 yielded an intermediate phenotype with responses to LPS diminished to a greater extent than that resulting from expression of the D299G TLR4 polymorphic variant. PMID- 18424734 TI - Mannose-binding lectin (MBL)-associated serine protease (MASP)-1 contributes to activation of the lectin complement pathway. AB - The complement system plays an important role in innate immunity. In the lectin complement pathway, mannose-binding lectin (MBL) and ficolins act as recognition molecules, and MBL-associated serine protease (MASP) is a key enzyme. It has been suggested that MASP-2 is responsible for the activation of C4. Other serine proteases (MASP-1 and MASP-3) are also associated with MBL or ficolins; however, their functions are still controversial. In this study, a MASP-1- and MASP-3 deficient mouse model (MASP1/3(-/-)) was generated by a gene targeting strategy to investigate the roles of MASP-1 and MASP-3 in the lectin pathway. Serum derived from MASP1/3(-/-) mice showed significantly lower activity of both C4 and C3 deposition on mannan-agarose, and this low activity was restored by the addition of recombinant MASP-1. MASP-1/3-deficient serum showed a significant delay for activation of MASP-2 compared with normal serum. Reconstitution of recombinant MASP-1 in MASP-1/3-deficient serum was able to promote the activation of MASP-2. From these results, we propose that MASP-1 contributes to the activation of the lectin pathway, probably through the activation of MASP-2. PMID- 18424733 TI - Single and dual amino acid substitutions in TCR CDRs can enhance antigen-specific T cell functions. AB - Single and dual amino acid substitution variants were generated in the TCR CDRs of three TCRs that recognize tumor-associated Ags. Substitutions that enhance the reactivity of TCR gene-modified T cells to the cognate Ag complex were identified using a rapid RNA-based transfection system. The screening of a panel of variants of the 1G4 TCR, that recognizes a peptide corresponding to amino acid residues 157-165 of the human cancer testis Ag NY-ESO-1 (SLLMWITQC) in the context of the HLA-A*02 class I allele, resulted in the identification of single and dual CDR3alpha and CDR2beta amino acid substitutions that dramatically enhanced the specific recognition of NY-ESO-1(+)/HLA-A*02(+) tumor cell lines by TCR gene modified CD4(+) T cells. Within this group of improved TCRs, a dual substitution in the 1G4 TCR CDR3alpha chain was identified that enhanced Ag-specific reactivity in gene-modified CD4(+) and CD8(+) T cells. Separate experiments on two distinct TCRs that recognize the MART-1 27-35 (AAGIGILTV) peptide/HLA-A*02 Ag complex characterized single amino acid substitutions in both TCRs that enhanced CD4(+) T cell Ag-specific reactivity. These results indicate that simple TCR substitution variants that enhance T cell function can be identified by rapid transfection and assay techniques, providing the means for generating potent Ag complex-specific TCR genes for use in the study of T cell interactions and in T cell adoptive immunotherapy. PMID- 18424735 TI - Immunological and genetic evidence for a crucial role of IL-10 in cutaneous lesions in humans infected with Leishmania braziliensis. AB - In populations exposed to Leishmania braziliensis, certain subjects develop skin ulcers, whereas others are naturally protected against cutaneous leishmaniasis. We have evaluated which cytokines are most crucial in the development of skin lesions. We found that active lesions occur in subjects with polarized Th2 or mixed Th1/Th2 responses, both associated with elevated IL-10 production. IL-10 was strongly associated (p = 0.004, odd ratio (OR) = 6.8, confidence interval = 1.9-25) with lesions, excluding IFN-gamma, IL-12, TNF, IL-13, and IL-4 from the regression model. IL-10 was produced by blood monocytes and CD4(+)CD25(+) T lymphocytes (mostly Foxp3(+)). However, we did not observe any difference between the number of these cells present in the blood of subjects with active lesions and those present in resistant subjects. Genetic analysis of the IL10-819C/T polymorphism, located in the IL10 promoter, showed that the C allele increased the risk of lesions (OR = 2.5 (1.12-5.7), p = 0.003). Functional analysis of these variants showed allele-specific binding of nuclear factors. The IL10-819C/C genotype was associated with higher levels of IL-10 than C/T and T/T genotypes. These observations demonstrate an important role for IL-10 in skin lesions in humans infected with L. braziliensis, and identify circulating monocytes and Tregs as principal sources of IL-10 in these patients. PMID- 18424736 TI - Lipoproteins are critical TLR2 activating toxins in group B streptococcal sepsis. AB - Group B streptococcus (GBS) is the most important cause of neonatal sepsis, which is mediated in part by TLR2. However, GBS components that potently induce cytokines via TLR2 are largely unknown. We found that GBS strains of the same serotype differ in released factors that activate TLR2. Several lines of genetic and biochemical evidence indicated that lipoteichoic acid (LTA), the most widely studied TLR2 agonist in Gram-positive bacteria, was not essential for TLR2 activation. We thus examined the role of GBS lipoproteins in this process by inactivating two genes essential for bacterial lipoprotein (BLP) maturation: the prolipoprotein diacylglyceryl transferase gene (lgt) and the lipoprotein signal peptidase gene (lsp). We found that Lgt modification of the N-terminal sequence called lipobox was not critical for Lsp cleavage of BLPs. In the absence of lgt and lsp, lipoprotein signal peptides were processed by the type I signal peptidase. Importantly, both the Deltalgt and the Deltalsp mutant were impaired in TLR2 activation. In contrast to released factors, fixed Deltalgt and Deltalsp GBS cells exhibited normal inflammatory activity indicating that extracellular toxins and cell wall components activate phagocytes through independent pathways. In addition, the Deltalgt mutant exhibited increased lethality in a model of neonatal GBS sepsis. Notably, LTA comprised little, if any, inflammatory potency when extracted from Deltalgt GBS. In conclusion, mature BLPs, and not LTA, are the major TLR2 activating factors from GBS and significantly contribute to GBS sepsis. PMID- 18424737 TI - A DNA vaccine prime followed by a liposome-encapsulated protein boost confers enhanced mucosal immune responses and protection. AB - A variety of DNA vaccine prime and recombinant viral boost immunization strategies have been developed to enhance immune responses in humans, but inherent limitations to these strategies exist. There is still an overwhelming need to develop safe and effective approaches that raise broad humoral and T cell mediated immune responses systemically and on mucosal surfaces. We have developed a novel mucosal immunization regimen that precludes the use of viral vectors yet induces potent T cell responses. Using hepatitis B surface Ag (HBsAg), we observed that vaccination of BALB/c mice with an i.m. HBsAg-DNA vaccine prime followed by an intranasal boost with HBsAg protein encapsulated in biologically inert liposomes enhanced humoral and T cell immune responses, particularly on mucosal surfaces. Intranasal live virus challenge with a recombinant vaccinia virus expressing HBsAg revealed a correlation between T cell immune responses and protection of immunized mice. A shortened immunization protocol was developed that was successful in both adult and neonatal mice. These results support the conclusion that this new approach is capable of generating a Th-type-1-biased, broad spectrum immune response, specifically at mucosal surfaces. The success of this design may provide a safe and effective vaccination alternative for human use. PMID- 18424738 TI - Alternatively activated myeloid cells limit pathogenicity associated with African trypanosomiasis through the IL-10 inducible gene selenoprotein P. AB - Uncontrolled inflammation is a major cause of tissue injury/pathogenicity often resulting in death of a host infected with African trypanosomes. Thus, comparing the immune response in hosts that develop different degrees of disease severity represents a promising approach to discover processes contributing to trypanosomiasis control. It is known that limitation of pathogenicity requires a transition in the course of infection, from an IFN-gamma-dependent response resulting in the development of classically activated myeloid cells (M1), to a counterbalancing IL-10-dependent response associated with alternatively activated myeloid cells (M2). Herein, mechanisms and downstream effectors by which M2 contribute to lower the pathogenicity and the associated susceptibility to African trypanosomiasis have been explored. Gene expression analysis in IL-10 knockout and wild-type mice, that are susceptible and relatively resistant to Trypanosoma congolense infection, respectively, revealed a number of IL-10 inducible genes expressed by M2, including Sepp1 coding for selenoprotein P. Functional analyses confirm that selenoprotein P contributes to limit disease severity through anti-oxidant activity. Indeed, Sepp1 knockout mice, but not Sepp1(Delta)(240-361) mice retaining the anti-oxidant motif but lacking the selenium transporter domain of selenoprotein P, exhibited increased tissue injury that associated with increased production of reactive oxygen species and increased apoptosis in the liver immune cells, reduced parasite clearance capacity of myeloid cells, and decreased survival. These data validate M2 associated molecules as functioning in reducing the impact of parasite infection on the host. PMID- 18424740 TI - Tick saliva inhibits dendritic cell migration, maturation, and function while promoting development of Th2 responses. AB - Similarly to other blood-feeding arthropods, ticks have evolved immunosuppressive mechanisms enabling them to overcome the host immune system. Although the immunomodulatory effect of tick saliva on several cell populations of the immune system has been extensively studied, little is known about its impact on dendritic cells (DCs). We have examined the effect of Ixodes ricinus tick saliva on DC function in vitro and in vivo. Exposure of DCs to tick saliva in vitro resulted in impaired maturation, upon CD40 or TLR9, TLR3 and TLR7 ligation, as well as reduced Ag presentation capacity. Administration of tick saliva in vivo significantly inhibited maturation and early migration of DCs from inflamed skin to draining lymph nodes, and decreased the capacity of lymph node DCs to present soluble Ag to specific T cells. Moreover, saliva-exposed DCs failed to induce efficient Th1 and Th17 polarization and promoted development of Th2 responses. Our data reveal a complex inhibitory effect exerted by tick saliva on DC function. Given the role of DCs as the key instigators of adaptive immune responses, alteration of their function might represent a major mechanism of tick mediated immune evasion. PMID- 18424739 TI - Neisseria gonorrhoeae-induced human defensins 5 and 6 increase HIV infectivity: role in enhanced transmission. AB - Sexually transmitted infections (STIs) increase the likelihood of HIV transmission. Defensins are part of the innate mucosal immune response to STIs and therefore we investigated their role in HIV infection. We found that human defensins 5 and 6 (HD5 and HD6) promoted HIV infection, and this effect was primarily during viral entry. Enhancement was seen with primary viral isolates in primary CD4(+) T cells and the effect was more pronounced with R5 virus compared with X4 virus. HD5 and HD6 promoted HIV reporter viruses pseudotyped with vesicular stomatitis virus and murine leukemia virus envelopes, indicating that defensin-mediated enhancement was not dependent on CD4 and coreceptors. Enhancement of HIV by HD5 and HD6 was influenced by the structure of the peptides, as loss of the intramolecular cysteine bonds was associated with loss of the HIV-enhancing effect. Pro-HD5, the precursor and intracellular form of HD5, also exhibited HIV-enhancing effect. Using a cervicovaginal tissue culture system, we found that expression of HD5 and HD6 was induced in response to Neisseria gonorrhoeae (GC, for gonococcus) infection and that conditioned medium from GC-exposed cervicovaginal epithelial cells with elevated levels of HD5 also enhanced HIV infection. Introduction of small interfering RNAs for HD5 or HD6 abolished the HIV-enhancing effect mediated by GC. Thus, the induction of these defensins in the mucosa in the setting of GC infection could facilitate HIV infection. Furthermore, this study demonstrates the complexity of defensins as innate immune mediators in HIV transmission and warrants further investigation of the mechanism by which defensins modulate HIV infection. PMID- 18424741 TI - T cell response mediated by myeloid cell-derived IL-12 is responsible for Porphyromonas gingivalis-induced periodontitis in IL-10-deficient mice. AB - Periodontal disease is a chronic inflammatory disease in the oral cavity, which culminates in alveolar bone loss. Porphyromonas gingivalis is a consensus periodontal pathogen that has been implicated in adult forms of periodontitis. We previously demonstrated that IL-10-deficient mice exhibit a hyperinflammatory phenotype and are highly susceptible to P. gingivalis-induced periodontitis, indicating an important anti-inflammatory effect of IL-10 in suppressing bone loss. In this study, we analyzed the pathway(s) by which IL-10 deficiency leads to severe P. gingivalis-induced periodontitis. Because Stat3 is essential in IL 10 signaling, immune cell-specific Stat3-deficient mice were subjected to P. gingivalis infection to identify the key IL-10-responsive cells in preventing periodontitis. Myeloid cell-specific Stat3-deficient mice exhibited increased periodontal bone loss (p < 0.001), whereas T cell- and B cell-specific Stat3 mice were resistant, suggesting that macrophages (MP) and/or polymorphonuclear leukocytes are the key target cells normally suppressed by IL-10. Myeloid cell specific Stat3-deficient mice exhibited elevated gingival CD40L gene expression in vivo compared with wild-type controls (p < 0.01), and Stat3-deficient MPs exhibited vigorous P. gingivalis-stimulated IL-12 production in vitro and induced elevated Ag-specific T cell proliferation compared with wild-type MPs (p < 0.01). Of importance, both IL-12p40/IL-10 and T cell/IL-10 double-deficient mice were resistant to P. gingivalis-induced periodontitis, demonstrating roles for both IL 12p40 and T cells in pathogenesis in a hyperinflammatory model of disease. These data demonstrate that P. gingivalis-induced periodontitis in IL-10-deficient mice is dependent upon IL-12p40-mediated proinflammatory T cell responses. PMID- 18424743 TI - Leukotriene B4 induces release of antimicrobial peptides in lungs of virally infected mice. AB - Leukotriene B(4) (LTB(4)) is a lipid mediator of inflammation that was recently shown to exert antiviral activities. In this study, we demonstrate that the release of antimicrobial proteins by neutrophils contribute to an early host defense against influenza virus infection in vitro as well as in vivo. Daily i.v. treatments with LTB(4) lead to a significant decrease in lung viral loads at day 5 postinfection in mice infected with influenza A virus compared with the placebo treated group. This reduction in viral load was not present in mice deficient in the high-affinity LTB(4) receptor. Viral clearance in lungs was associated with up-regulated presence of antimicrobial peptides such as beta-defensin-3, members of the mouse eosinophil-related RNase family, and the mouse cathelicidin-related antimicrobial peptide. Our results also indicate that neutrophils are important in the antiviral effect of LTB(4). Viral loads in neutrophil-depleted mice were not diminished by LTB(4) administration, and a substantial reduction in the presence of murine cathelicidin-related antimicrobial peptide and the murine eosinophil-related RNase family in lung tissue was observed. Moreover, in vitro treatment of human neutrophil cultures with LTB(4) led rapidly to the secretion of the human cathelicidin LL-37 and eosinophil-derived neurotoxin, known as antiviral peptides. Pretreatment of cell cultures with specific LTB(4) receptor antagonists clearly demonstrate the implication of the high-affinity LTB(4) receptor in the LTB(4)-mediated activity. Together, these results demonstrate the importance of neutrophils and the secretion of antimicrobial peptides during the early immune response mediated by LTB(4) against a viral pathogen. PMID- 18424742 TI - Inhibition of Yin Yang 1-dependent repressor activity of DR5 transcription and expression by the novel proteasome inhibitor NPI-0052 contributes to its TRAIL enhanced apoptosis in cancer cells. AB - TRAIL promotes apoptotic tumor cell death; however, TRAIL-resistant tumors need to be sensitized to reverse resistance. Proteasome inhibitors potentiate TRAIL apoptosis in vitro and in vivo and correlate with up-regulation of death receptor 5 (DR5) via an unknown mechanism. We hypothesized that the proteasome inhibitor NPI-0052 inhibits the transcription repressor Yin Yang 1 (YY1) which regulates TRAIL resistance and negatively regulates DR5 transcription. Treatment of PC-3 and Ramos cells with NPI-0052 (A; p.Arg240His), which is associated with aHUS. This heterozygous change was found in 6/166 aHUS patients compared with 5/542 normal (chi2 = 6.021; p = 0.014), which was replicated in a second cohort of aHUS patients in which we found 5/170 carriers. The polymorphism does not decrease expression efficiency of C4BP. p.Arg240His is equally efficient as the wild type in binding and supporting degradation of C4BP but its ability to bind C3b and act as cofactor to its degradation both in fluid phase and on surfaces is impaired. This observation supports the hypothesis that dysregulation of the alternative pathway of complement is pivotal for aHUS. Three of the patients carry also mutations in membrane cofactor protein and factor H strengthening the hypothesis that individuals may carry multiple susceptibility factors with an additive effect on the risk of developing aHUS. PMID- 18424764 TI - Correlation of blood T cell and antibody reactivity to myelin proteins with HLA type and lesion localization in multiple sclerosis. AB - Multiple sclerosis (MS) is a chronic inflammatory demyelinating disease of the CNS. The numbers of autoimmune T cells and Abs specific for proteins of CNS myelin are increased in the blood in some patients with MS. The aim of this study was to investigate whether there are correlations between the specificity of the autoimmune responses in the blood, the HLA molecules carried by the patient, and the clinical features of MS, because studies on experimental autoimmune encephalomyelitis, an animal model of MS, indicate that autoimmune responses targeting particular myelin proteins and the genetic background of the animal play a role in determining the pattern of lesion distribution. We tested blood T cell immunoreactivity to myelin proteins in 100 MS patients, 70 healthy controls, and 48 patients with other neurological disorders. Forty MS patients had strongly increased T cell reactivity to one or more myelin Ags. In these 40 patients, the most robust correlation was between CD4(+) T cell reactivity to myelin proteolipid protein residues 184-209 (PLP(184-209)) and development of lesions in the brainstem and cerebellum. Furthermore, carriage of HLA-DR4, -DR7, or -DR13 molecules by MS patients correlated with increased blood T cell immunoreactivity to PLP(184-209), as well as the development of lesions in the brainstem and cerebellum. Levels of PLP(190-209)-specific Abs in the blood also correlated with the presence of cerebellar lesions. These findings show that circulating T cells and Abs reactive against specific myelin Ags can correlate with lesion distribution in MS and suggest that they are of pathogenic relevance. PMID- 18424763 TI - Blocking NK cell inhibitory self-recognition promotes antibody-dependent cellular cytotoxicity in a model of anti-lymphoma therapy. AB - Human NK cells lyse Ab-coated target cells through the process of Ab-dependent cellular cytotoxicity (ADCC). Improving ADCC responses is desirable because it is thought to be an important antitumor mechanism for some Abs. NK cell inhibitory receptors, such as killer cell Ig-like receptors, engage with MHC class I molecules on self-cells to block NK cell activation. Accordingly, we enhanced ADCC responses by blocking NK cell inhibitory receptors, thus perturbing induction of the self-recognition signal. In a cell line model of anti-lymphoma therapy, the combination of rituximab with an Ab that blocks inhibitory self recognition yielded increased NK cell-mediated target cell lysis when compared with rituximab alone. To validate this proof-of-concept, we then used a more representative approach in which an individual's fresh primary NK cells encountered autologous, EBV-transformed B cells. In this system, rituximab and a combination of Abs that block NK cell inhibitory receptors yielded improved NK cell-mediated lysis over rituximab alone. The results show, for the first time, that disruption of inhibitory self-recognition can efficiently promote ADCC in a human model, applying an autologous system in which physiologic checkpoints are in place. This method provides an alternative approach to potentiate the therapeutic benefit of antitumor Abs that mediate ADCC. PMID- 18424765 TI - Natural naive CD4+CD25+CD127low regulatory T cell (Treg) development and function are disturbed in multiple sclerosis patients: recovery of memory Treg homeostasis during disease progression. AB - Patients with relapsing-remitting multiple sclerosis (RR-MS) show a suboptimal CD4(+)CD25(+) regulatory T cell (Treg) function, whereas no Treg alterations are observed in secondary progressive MS (SP-MS) patients. To clarify the difference in Treg activity between early and chronic disease stages in MS, we analyzed the functional capacity and homeostatic parameters of naive CD4(+)CD25(+)CD127(low)CD45RA(+) Tregs (nTregs) and their memory counterparts CD4(+)CD25(+)CD127(low)CD45RO(+) Tregs (mTregs) in untreated MS patients and healthy controls. Interestingly, whereas the suppressive capacity of FACS-sorted nTregs was impaired in both early and chronic MS patients, only the latter group showed a restored mTreg function. Consistent with this observation, chronic MS patients had increased numbers of mTregs as compared with age-matched early MS patients, whereas nTreg frequencies did not differ significantly. TCR excision circle numbers were reduced in nTregs of early MS patients, suggestive of a diminished nTreg thymic output. Moreover, a decreased number of CD31(+) mTregs were observed in early vs chronic MS patients, indicating that inflammatory processes drive the homeostatic turnover of mTregs during the early disease stage. Additionally, early MS patients showed a more restricted nTreg and mTreg TCR BV gene profile as compared with healthy controls and chronic MS patients. Finally, analysis of IFN-beta and glatiramer acetate-treated MS patients showed that these immunomodulatory drugs modify nTreg homeostasis. Taken together, this study provides strong evidence for a disturbed thymic nTreg development and function in MS patients. Moreover, memory Treg but not naive Treg homeostasis recovers during disease progression. PMID- 18424766 TI - Decreased expression of the fractalkine receptor CX3CR1 on circulating monocytes as new feature of sepsis-induced immunosuppression. AB - Although it is known that septic shock rapidly induces immune dysfunctions, which contribute to the impaired clearance of microorganisms observed in patients, the mechanisms for this phenomenon remain incompletely understood. We recently observed, in a microarray study, an altered circulating leukocyte CX3CR1 mRNA expression associated with patients' mortality. As monocytes play a central role in septic shock pathophysiology and express high levels of CX3CR1, we therefore further investigated the alteration of CX3CR1 expression and of its ligand fractalkine (CX3CL1) on those cells in this clinical condition. We observed that CX3CR1 expression (both mRNA and protein) was severely down-regulated in monocytes and consequently associated with a lack of functionality upon fractalkine challenge. Importantly, nonsurvivors presented with significantly sustained lower expression in comparison with survivors. This down-regulation was reproduced by incubation of cells from healthy individuals with LPS, whole bacteria (Escherichia coli and Staphylococcus aureus), and, to a lower extent, with corticosteroids-in accordance with the concept of LPS-induced monocyte deactivation. In addition, CX3CL1 serum concentrations were elevated in patients supporting the hypothesis of increased cleavage of the membrane-anchored form expressed by endothelial cells. As CX3CR1/CX3CL1 interaction preferentially mediates arrest and migration of proinflammatory cells, the present observations may contribute to patients' inability to kill invading microorganisms. This could represent an important new feature of sepsis-induced immunosuppression. PMID- 18424767 TI - CREB has a context-dependent role in activity-regulated transcription and maintains neuronal cholesterol homeostasis. AB - Induction of specific gene expression patterns in response to activity confers functional plasticity to neurons. A principal role in the regulation of these processes has been ascribed to the cAMP responsive element binding protein (CREB). Using genome-wide expression profiling in mice lacking CREB in the forebrain, accompanied by deletion of the cAMP responsive element modulator gene (CREM), we here show that the role of these proteins in activity-induced gene expression is surprisingly selective and highly context dependent. Thus, only a very restricted subset of activity-induced genes (i.e., Gadd45b or Nr4a2) requires these proteins for their induction in the hippocampus after kainic acid administration, while they are required for most of the cocaine-induced expression changes in the striatum. Interestingly, in the absence of CREB, CREM is able to rescue activity-regulated transcription, which strengthens the notion of overlapping functions of the two proteins. In addition, we show that cholesterol metabolism is dysregulated in the brains of mutant mice, as reflected coordinated expression changes in genes involved in cholesterol synthesis and neuronal accumulation of cholesterol. These findings provide novel insights into the role of CREB and CREM in stimulus-dependent transcription and neuronal homeostasis. PMID- 18424768 TI - A protein complex in the brush-border membrane explains a Hartnup disorder allele. AB - Protein absorption in the intestine is mediated by proteases and brush-border peptidases together with peptide and amino acid transporters. Neutral amino acids are generated by a variety of aminopeptidases and carboxypeptidases and are subsequently taken up by the amino acid transporter B(0)AT1 (SLC6A19), which is mutated in Hartnup disorder. Coexpression of B(0)AT1 together with the brush border carboxypeptidase angiotensin-converting enzyme 2 (ACE2) in Xenopus laevis oocytes led to a dramatic increase of transporter expression at the oocyte surface. Other members of the SLC6 family were not stimulated by coexpression with ACE2. Addition of a peptide containing a carboxyterminal leucine residue to ACE2- and B(0)AT1-coexpressing oocytes caused inward currents due to Na(+) leucine cotransport, demonstrating the formation of a metabolic complex. Coexpression of the Hartnup disorder causing mutation B(0)AT1(R240Q) showed reduced interaction with ACE2 and its renal paralogue collectrin. This would result in reduced surface expression in both kidney and intestine, thereby explaining the onset of the disorder in individuals carrying this mutation. PMID- 18424769 TI - Gamma-secretase and metalloproteinase activity regulate the distribution of endoplasmic reticulum to hippocampal neuron dendritic spines. AB - The neuronal endoplasmic reticulum (ER) contributes to many physiological and pathological processes in the brain. A subset of dendritic spines on hippocampal neurons contains ER that may contribute to synapse-specific intracellular signaling. Distribution of ER to spines is dynamic, but knowledge of the regulatory mechanisms is lacking. In live cell imaging experiments we now show that cultured hippocampal neurons rapidly lost ER from spines after phorbol ester treatment. ER loss was reduced by inhibiting gamma-secretase (DAPT at 2 microM) and metalloproteinase (TAPI-0 and GM6001 at 4 microM) activity. Inhibition of protein kinase C also diminished loss of ER by preventing exit of ER from spines. Furthermore, gamma-secretase and metalloproteinase inhibition, in the absence of phorbol ester, triggered a dramatic increase in spine ER content. Metalloproteinases and gamma-secretase cleave several transmembrane proteins. Many of these substrates are known to localize to adherens junctions, a structural specialization with which spine ER interacts. One interesting possibility is thus that ER content within spines may be regulated by proteolytic activity affecting adherens junctions. Our data demonstrate a hitherto unknown role for these two proteolytic activities in regulating dynamic aspects of cellular ultrastructure, which is potentially important for cellular calcium homeostasis and several intracellular signaling pathways. PMID- 18424770 TI - The Wilms' tumor suppressor Wt1 activates transcription of the erythropoietin receptor in hematopoietic progenitor cells. AB - The Wilms' tumor protein Wt1 is required for embryonic development and has been implicated in hematologic disorders. Since Wt1 deficiency may compromise the proliferation and differentiation of erythroid progenitor cells, we analyzed the possible role of the transcriptionally active Wt1 isoform, Wt1(-KTS), in regulating the expression of the erythropoietin receptor (EpoR). Wt1 and EpoR were coexpressed in CD117(+) hematopoietic progenitor cells and in several hematopoietic cell lines. CD117(+) cells of Wt1-deficient murine embryos (Wt1(-/ )) exhibited a significantly lower proliferation response to recombinant erythropoietin than CD117(+) cells of heterozygous (Wt1(+/-)) and wild-type littermates (Wt1(+/+)). EpoR expression was significantly diminished in hematopoietic progenitors (CD117(+)) that lacked Wt1, and the erythroid colony forming capacity was reduced by more than 50% in fetal liver cells of Wt1 deficient embryonic mice. Wt1(-KTS) significantly increased endogenous EpoR transcripts in transfected cells. The proximal EpoR promoter of human and mouse was stimulated more than 10-fold by Wt1(-KTS) in transiently cotransfeced K562 erythroleukemia cells. A responsible cis-element, which is highly conserved in the EpoR promoter of human and mouse, was identified by mutation analysis, electrophoretic mobility shift assay, and chromatin immunoprecipitation assay. In conclusion, activation of the EpoR gene by Wt1 may represent an important mechanism in normal hematopoiesis. PMID- 18424771 TI - Evaluating phonological processing skills in children with prelingual deafness who use cochlear implants. AB - This study investigated the phonological processing skills of 29 children with prelingual, profound hearing loss with 4 years of cochlear implant experience. Results were group matched with regard to word-reading ability and mother's educational level with the performance of 29 hearing children. Results revealed that it is possible to obtain a valid measure of phonological processing (PP) skills in children using CIs. They could complete rhyming tasks and were able to complete sound-based tasks using standard test materials provided by a commercial test distributor. The CI children completed tasks measuring PP, but there were performance differences between the CI users and the hearing children. The process of learning phonological awareness (PA) for the children with CIs was characterized by a longer, more protracted learning phase than their counterparts with hearing. Tests of phonological memory skills indicated that when the tasks were controlled for presentation method and response modality, there were no differences between the performance of children with CIs and their counterparts with hearing. Tests of rapid naming revealed that there were no differences between rapid letter and number naming between the two groups. Results yielded a possible PP test battery for children with CI experience. PMID- 18424772 TI - Social construction of American sign language--English interpreters. AB - Instructors in 5 American Sign Language--English Interpreter Programs and 4 Deaf Studies Programs in Canada were interviewed and asked to discuss their experiences as educators. Within a qualitative research paradigm, their comments were grouped into a number of categories tied to the social construction of American Sign Language--English interpreters, such as learners' age and education and the characteristics of good citizens within the Deaf community. According to the participants, younger students were adept at language acquisition, whereas older learners more readily understood the purpose of lessons. Children of deaf adults were seen as more culturally aware. The participants' beliefs echoed the theories of P. Freire (1970/1970) that educators consider the reality of each student and their praxis and were responsible for facilitating student self awareness. Important characteristics in the social construction of students included independence, an appropriate attitude, an understanding of Deaf culture, ethical behavior, community involvement, and a willingness to pursue lifelong learning. PMID- 18424773 TI - Difficulties in testing for covarion-like properties of sequences under the confounding influence of changing proportions of variable sites. AB - The covarion (COV)-like properties of sequences are poorly described and their impact on phylogenetic analyses poorly understood. We demonstrate using simulations that, under an evolutionary model where the proportion of variable sites changes in nonadjacent lineages, log likelihood values for rates across site (RAS) and COV models become similar, making models difficult to distinguish. Further, although COV and RAS models provide a great improvement in likelihood scores over a homogeneous model with these simulated data, reconstruction accuracy of tree building is low, suggesting caution when it is suspected that proportions of variable sites differ in different evolutionary lineages. We study the performance of a recently developed contingency test that detects the presence of COV-type evolution modified for protein data. We report that if proportions of variable sites (p(var)) change in a lineage-specific manner such that their distributions in different lineages become sufficiently nonoverlapping, then the contingency test can incorrectly suggest a homogeneous model. Also of concern is the possibility of different proportions of variable sites between the groups being studied. In a study of chloroplast proteins, interpretation of the test is found to be susceptible to different partitioning of taxon groups, making the test very subjective in its implementation. Extreme intergroup differences in the extent of divergence and difference in proportions of variable sites could be contributing to this effect. PMID- 18424774 TI - Differential evolution of the 13 Atlantic salmon Hox clusters. AB - Hox cluster organization represents a valuable marker to study the effects of recent genome duplication in salmonid fish (25-100 Mya). Using polymerase chain reaction amplification of cDNAs, BAC library screening, and genome walking, we reconstructed 13 Hox clusters in the Atlantic salmon containing 118 Hox genes including 8 pseudogenes. Hox paralogs resulting from the genome duplication preceding the radiation of ray-finned fish have been much better preserved in salmon than in other model teleosts. The last genome duplication in the salmon lineage has been followed by the loss of 1 of the 4 HoxA clusters. Four rounds of genome duplication after the vertebrate ancestor salmon Hox clusters display the main organizational features of vertebrate Hox clusters, with Hox genes exclusively that are densely packed in the same orientation. Recently, duplicated Hox clusters have engaged a process of divergence, with several cases of pseudogenization or asymmetrical evolution of Hox gene duplicates, and a marked erosion of identity in noncoding sequences. Strikingly, the level of divergence attained strongly depends on the Hox cluster pairs rather than on the Hox genes within each cluster. It is particularly high between both HoxBb clusters and both HoxDa clusters, whereas both HoxBa clusters remained virtually identical. Positive selection on the Hox protein-coding sequences could not be detected. PMID- 18424775 TI - Where is a nose with respect to a foot? The left posterior parietal cortex processes spatial relationships among body parts. AB - Neuropsychological studies suggest that patients with left parietal lesions may show impaired localization of parts of either their own or the examiner's body, despite preserved ability to identify isolated body parts. This deficit, called autotopagnosia, may result from damage to the Body Structural Description (BSD), a representation which codes spatial relationships among body parts. We used functional magnetic resonance imaging to identify the neural mechanisms underlying the BSD. Two human body or building parts (factor: STIMULI) were shown to participants who either identified them or evaluated their distance (factor: TASK). The analysis of the interaction between STIMULI and TASK, which isolates the neural mechanism underlying BSD, revealed an activation of left posterior intraparietal sulcus (IPS) when the distance between body parts was evaluated. The results show that the left IPS processes specifically the information about spatial relationships among body parts and thereby suggest that damage to this area may underlie autotopagnosia. PMID- 18424776 TI - Language conflict in the bilingual brain. AB - The large majority of humankind is more or less fluent in 2 or even more languages. This raises the fundamental question how the language network in the brain is organized such that the correct target language is selected at a particular occasion. Here we present behavioral and functional magnetic resonance imaging data showing that bilingual processing leads to language conflict in the bilingual brain even when the bilinguals' task only required target language knowledge. This finding demonstrates that the bilingual brain cannot avoid language conflict, because words from the target and nontarget languages become automatically activated during reading. Importantly, stimulus-based language conflict was found in brain regions in the LIPC associated with phonological and semantic processing, whereas response-based language conflict was only found in the pre-supplementary motor area/anterior cingulate cortex when language conflict leads to response conflicts. PMID- 18424777 TI - Auditory, somatosensory, and multisensory insular cortex in the rat. AB - Compared with other areas of the forebrain, the function of insular cortex is poorly understood. This study examined the unisensory and multisensory function of the rat insula using high-resolution, whole-hemisphere, epipial evoked potential mapping. We found the posterior insula to contain distinct auditory and somatotopically organized somatosensory fields with an interposed and overlapping region capable of integrating these sensory modalities. Unisensory and multisensory responses were uninfluenced by complete lesioning of primary and secondary auditory and somatosensory cortices, suggesting a high degree of parallel afferent input from the thalamus. In light of the established connections of the posterior insula with the amygdala, we propose that integration of auditory and somatosensory modalities reported here may play a role in auditory fear conditioning. PMID- 18424779 TI - Selective attention increases the dependency of cortical responses on visual motion coherence in man. AB - Attention improves visual discrimination and consequently allows to discern stimuli with low signal-to-noise ratios that otherwise would remain undetected. We used magnetoencephalography (MEG) to test whether neuromagnetic responses recorded from occipito-temporal cortex, reflecting the size of visual motion signals embedded in noise (motion coherence), would mirror the perceptual changes induced by attention. Attention directed to a given hemifield increased and decreased the coherence modulation of the MEG response over contralateral and ipsilateral visual cortex, respectively, indicating a change in the neuronal signal-to-noise ratio at the population level. PMID- 18424778 TI - An additional motor-related field in the lateral frontal cortex of squirrel monkeys. AB - Our earlier efforts to document the cortical connections of the ventral premotor cortex (PMv) revealed dense connections with a field rostral and lateral to PMv, an area we called the frontal rostral field (FR). Here, we present data collected in FR using electrophysiological and anatomical methods. Results show that FR contains an isolated motor representation of the forelimb that can be differentiated from PMv based on current thresholds and latencies to evoke electromyographic activity using intracortical microstimulation techniques. In addition, FR has a different pattern of cortical connections compared with PMv. Together, these data support that FR is an additional, previously undescribed motor-related area in squirrel monkeys. PMID- 18424780 TI - Right parietal brain activity precedes perceptual alternation of bistable stimuli. AB - Momentary fluctuations of baseline activity have been shown to influence responses to sensory stimulation both behaviorally and neurophysiologically. This suggests that perceptual awareness does not solely arise from physical stimulus properties. Here we studied whether the momentary state of the brain immediately before stimulus presentation indicates how a physically unique but perceptually ambiguous stimulus will be perceived. A complex Necker cube was intermittently presented and subjects indicated whether their perception changed with respect to the preceding presentation. EEG was recorded from 256 channels. The prestimulus brain-state was defined as the spatial configuration of the scalp potential map within the 50 ms before stimulus arrival, representing the sum of all momentary ongoing brain processes. Two maps were found that doubly dissociated perceptual reversals from perceptual stability. For EEG sweeps classified as either map, distributed inverse solutions were computed and statistically compared. This yielded activity confined to a region in right inferior parietal cortex that was significantly more active before a perceptual reversal. In contrast, no significant topographic differences of the evoked potentials elicited by stable vs. reversed Necker cubes were found. This indicates that prestimulus activity in right inferior parietal cortex is associated with the perceptual change. PMID- 18424781 TI - Goodpasture antigen-binding protein and its spliced variant, ceramide transfer protein, have different functions in the modulation of apoptosis during zebrafish development. AB - Human Goodpasture antigen-binding protein (GPBP) is an atypical protein kinase that phosphorylates the Goodpasture auto-antigen, the alpha3 chain of collagen IV. The COL4A3BP gene is alternatively spliced producing two protein isoforms: GPBP and GPBPDelta26. The latter lacks a serine-rich domain composed of 26 amino acid residues. Both isoforms also function as ceramide transfer proteins (CERT). Here, we explored the function of Gpbp and GpbpDelta26/CERT during embryogenesis in zebrafish. We cloned both splice variants of the zebrafish gene and found that they are differentially expressed during development. We used antisense oligonucleotide-mediated loss-of-function and synthetic mRNA-based gain-of function approaches. Our results show that the loss-of-function phenotype is linked to cell death, evident primarily in the muscle of the somites, extensive loss of myelinated tracks, and brain edema. These results indicate that disruption of the nonvesicular ceramide transport is detrimental to normal embryonic development of somites and brain because of increased apoptosis. Moreover, this phenotype is mediated by Gpbp but not GpbpDelta26/CERT, suggesting that Gpbp is an important factor for normal skeletal muscle and brain development. PMID- 18424782 TI - A common beta-sheet architecture underlies in vitro and in vivo beta2 microglobulin amyloid fibrils. AB - Misfolding and aggregation of normally soluble proteins into amyloid fibrils and their deposition and accumulation underlies a variety of clinically significant diseases. Fibrillar aggregates with amyloid-like properties can also be generated in vitro from pure proteins and peptides, including those not known to be associated with amyloidosis. Whereas biophysical studies of amyloid-like fibrils formed in vitro have provided important insights into the molecular mechanisms of amyloid generation and the structural properties of the fibrils formed, amyloidogenic proteins are typically exposed to mild or more extreme denaturing conditions to induce rapid fibril formation in vitro. Whether the structure of the resulting assemblies is representative of their natural in vivo counterparts, thus, remains a fundamental unresolved issue. Here we show using Fourier transform infrared spectroscopy that amyloid-like fibrils formed in vitro from natively folded or unfolded beta(2)-microglobulin (the protein associated with dialysis-related amyloidosis) adopt an identical beta-sheet architecture. The same beta-strand signature is observed whether fibril formation in vitro occurs spontaneously or from seeded reactions. Comparison of these spectra with those of amyloid fibrils extracted from patients with dialysis-related amyloidosis revealed an identical amide I' absorbance maximum, suggestive of a characteristic and conserved amyloid fold. Our results endorse the relevance of biophysical studies for the investigation of the molecular mechanisms of beta(2) microglobulin fibrillogenesis, knowledge about which may inform understanding of the pathobiology of this protein. PMID- 18424783 TI - Inflammatory levels of nitric oxide inhibit airway epithelial cell migration by inhibition of the kinase ERK1/2 and activation of hypoxia-inducible factor-1 alpha. AB - Increased synthesis of NO during airway inflammation, caused by induction of nitric-oxide synthase 2 in several lung cell types, may contribute to epithelial injury and permeability. To investigate the consequence of elevated NO production on epithelial function, we exposed cultured monolayers of human bronchial epithelial cells to the NO donor diethylenetriaamine NONOate. At concentrations generating high nanomolar levels of NO, representative of inflammatory conditions, diethylenetriaamine NONOate markedly reduced wound closure in an in vitro scratch injury model, primarily by inhibiting epithelial cell migration. Analysis of signaling pathways and gene expression profiles indicated a rapid induction of the mitogen-activated protein kinase phosphatase (MPK)-1 and decrease in extracellular signal-regulated kinase (ERK)1/2 activation, as well as marked stabilization of hypoxia-inducible factor (HIF)-1alpha and activation of hypoxia-responsive genes, under these conditions. Inhibition of ERK1/2 signaling using U0126 enhanced HIF-1alpha stabilization, implicating ERK1/2 dephosphorylation as a contributing mechanism in NO-mediated HIF-1alpha activation. Activation of HIF-1alpha by the hypoxia mimic cobalt chloride, or cell transfection with a degradation-resistant HIF-1alpha mutant construct inhibited epithelial wound repair, implicating HIF-1alpha in NO-mediated inhibition of cell migration. Conversely, NO-mediated inhibition of epithelial wound closure was largely prevented after small interfering RNA suppression of HIF-1alpha. Finally, NO-mediated inhibition of cell migration was associated with HIF-1alpha-dependent induction of PAI-1 and activation of p53, both negative regulators of epithelial cell migration. Collectively, our results demonstrate that inflammatory levels of NO inhibit epithelial cell migration, because of suppression of ERK1/2 signaling, and activation of HIF-1alpha and p53, with potential consequences for epithelial repair and remodeling during airway inflammation. PMID- 18424785 TI - Estimating time-to-event from longitudinal ordinal data using random-effects Markov models: application to multiple sclerosis progression. AB - Longitudinal ordinal data are common in many scientific studies, including those of multiple sclerosis (MS), and are frequently modeled using Markov dependency. Several authors have proposed random-effects Markov models to account for heterogeneity in the population. In this paper, we go one step further and study prediction based on random-effects Markov models. In particular, we show how to calculate the probabilities of future events and confidence intervals for those probabilities, given observed data on the ordinal outcome and a set of covariates, and how to update them over time. We discuss the usefulness of depicting these probabilities for visualization and interpretation of model results and illustrate our method using data from a phase III clinical trial that evaluated the utility of interferon beta-1a (trademark Avonex) to MS patients of type relapsing-remitting. PMID- 18424784 TI - Bone morphogenetic protein 2 mediates dentin sialophosphoprotein expression and odontoblast differentiation via NF-Y signaling. AB - Dentin sialophosphoprotein (DSPP), an important odontoblast differentiation marker, is necessary for tooth development and mineralization. Bone morphogenetic protein 2 (BMP2) plays a vital role in odontoblast function via diverse signal transduction systems. We hypothesize that BMP2 regulates DSPP gene transcription and thus odontoblast differentiation. Here we report that expression of BMP2 and DSPP is detected during mouse odontogenesis by in situ hybridization assay, and BMP2 up-regulates DSPP mRNA and protein expression as well as DSPP-luciferase promoter activity in mouse preodontoblasts. By sequentially deleting fragments of the mouse DSPP promoter, we show that a BMP2-response element is located between nucleotides -97 and -72. By using antibody and oligonucleotide competition assays in electrophoretic mobility shift analysis and chromatin immunoprecipitation experiments, we show that the heterotrimeric transcription factor Y (NF-Y) complex physically interacts with the inverted CCAAT box within the BMP2-response element. BMP2 induces NF-Y accumulation into the nucleus increasing its recruitment to the mouse DSPP promoter in vivo. Furthermore, forced overexpression of NF-Y enhances promoter activity and increases endogenous DSPP protein levels. In contrast, mutations in the NF-Y-binding motif reduce BMP2 induced DSPP transcription. Moreover, inhibiting BMP2 signaling by Noggin, a BMP2 antagonist, results in significant inhibition of DSPP gene expression in preodontoblasts. Taken together, these results indicate that BMP2 mediates DSPP gene expression and odontoblast differentiation via NF-Y signaling during tooth development. PMID- 18424786 TI - Diabetes, left ventricular systolic dysfunction, and chronic heart failure. AB - Chronic heart failure (HF) and diabetes mellitus (DM) commonly coexist. Each condition increases the likelihood of developing the other, and when they occur together in the same patient the risk of morbidity and mortality increases markedly. We discuss the epidemiological overlap and consider the complex patho physiological pathways linking the two diseases. The treatment of each condition is made more problematic by the presence of the other. We review the evidence based treatment strategies and discuss the common problems faced by physicians when treating patients with both conditions. This article forms a comprehensive overview of a fascinating intersection between two common diseases. PMID- 18424787 TI - Sex differences in atheroma burden and endothelial function in patients with early coronary atherosclerosis. AB - AIMS: Women and men have different clinical presentations and outcomes in coronary artery disease (CAD). We tested the hypothesis that sex differences may influence coronary atherosclerotic burden and coronary endothelial function before development of obstructive CAD. METHODS AND RESULTS: A total of 142 patients (53 men, 89 women; mean +/- SD age, 49.3 +/- 11.7 years) with early CAD simultaneously underwent intravascular ultrasonography and coronary endothelial function assessment. Atheroma burden in the left main and proximal left anterior descending (LAD) arteries was significantly greater in men than women (median, 23.0% vs. 14.1%, P = 0.002; median, 40.1% vs. 29.3%, P = 0.001, respectively). Atheroma eccentricity in the proximal LAD artery was significantly higher in men than women (median, 0.89 vs. 0.80, P = 0.04). The length of the coronary segments with endothelial dysfunction was significantly longer in men than women (median, 39.2 vs. 11.1 mm, P = 0.002). In contrast, maximal coronary flow reserve was significantly lower in women than men (2.80 vs. 3.30, P < 0.001). Sex was an independent predictor of atheroma burden in the left main and proximal LAD arteries (both P < 0.05) by multivariate analysis. CONCLUSION: Men have greater atheroma burden, more eccentric atheroma, and more diffuse epicardial endothelial dysfunction than women. These results suggest that men have more severe structural and functional abnormalities in epicardial coronary arteries than women, even in patients with early atherosclerosis, which may result in the higher incidence rates of CAD and ST-segment myocardial infarction in men than women. PMID- 18424788 TI - Far-field oversensing of atrial signals: an unusual cause for very short V-V intervals and inappropriate implantable cardioverter defibrillator therapy. PMID- 18424789 TI - Fosfomycin tromethamine as second agent for the treatment of acute, uncomplicated urinary tract infections in adult female patients in The Netherlands? AB - BACKGROUND: Uncomplicated urinary tract infections (UTIs) are common among female patients. According to the national guidelines of the Dutch College of General Practitioners (GPs), the drugs of first and second choice as therapy for UTIs are nitrofurantoin and trimethoprim with resistance percentages of 2% and 23%, respectively. The third choice is fosfomycin tromethamine for which no current resistance data from The Netherlands are available. The aim of this study was to determine these resistance percentages. METHODS: During 2003-04, urine samples were collected from a representative sample of 21 general practices spread over The Netherlands, the Sentinel Stations of The Netherlands Institute for Health Services Research (NIVEL). Escherichia coli isolated from female patients visiting their GP with symptoms of an acute, uncomplicated UTI were used. Fosfomycin tromethamine susceptibility was determined by Etests. An MIC of fosfomycin tromethamine of 64 mg/L or lower was considered to indicate susceptibility, and MIC values of 96 mg/L or higher were considered to indicate resistance. E. coli ATCC 25922 was used as a reference strain. RESULTS: In total, 1705 E. coli strains were tested, of which 11 (0.65%) were resistant to fosfomycin tromethamine. The MIC(50) and MIC(90) values for this population were 1 and 4 mg/L, respectively. Within the inhibition zone of 162 susceptible E. coli, resistant mutant colonies were observed, of which after repetition of the susceptibility testing 68 were resistant. In total, 79 (5%) strains were resistant to fosfomycin tromethamine. There was no cross-resistance observed between fosfomycin tromethamine and other antimicrobial agents tested previously. CONCLUSIONS: The high in vitro susceptibility to fosfomycin tromethamine in this population and the lack of cross-resistance between fosfomycin tromethamine and other agents together with the extensive global clinical experience support the choice of the national guidelines of the Dutch College of GPs to include fosfomycin tromethamine as a therapeutic option in general practice for uncomplicated UTIs. PMID- 18424790 TI - In vitro activity of tigecycline against multidrug-resistant Acinetobacter baumannii. PMID- 18424791 TI - Incidence and antimicrobial susceptibilities of genital mycoplasmas in outpatient women with clinical vaginitis in Athens, Greece. AB - OBJECTIVES: The incidence and antimicrobial susceptibilities of Ureaplasma urealyticum and Mycoplasma hominis, isolated from vaginal and endocervical swabs collected from 369 outpatient women, were determined. METHODS: Isolation, identification and typing of the pathogens were performed by means of conventional methods. The antimicrobial susceptibilities of the genital mycoplasmas were determined with commercially available kits and evaluated according to the CLSI. RESULTS AND CONCLUSIONS: In 65 (47.44%) out of the 137 positive specimens, U. urealyticum was grown as a single pathogen, in 0.72% M. hominis was grown as a single pathogen and in 2.92% both urogenital mycoplasmas were grown. In the remaining specimens (48.90%), there was a mixed growth with other microbes. Of the isolated U. urealyticum strains, 87.4% and 98.2% were susceptible to tetracycline and doxycycline, respectively, 79.2% were susceptible to josamycin, 48.6% were susceptible to clarithromycin and 91.8% were susceptible to pristinamycin, while erythromycin, azithromycin, ciprofloxacin and ofloxacin proved to be inactive against most of the strains. M. hominis isolates were 100% susceptible to tetracycline, doxycycline and pristinamycin, while susceptibilities to the other antimicrobial agents varied mainly in the range of 'intermediate' or 'resistant'. As results originating from similar studies from various countries are very controversial, the simplest way to avoid therapeutic failures would be the implementation of rational treatment regimens based on culture isolation and the in vitro determination of the antimicrobial susceptibility of genital mycoplasmas in each clinical case. PMID- 18424792 TI - In vitro activity of telavancin against recent Gram-positive clinical isolates: results of the 2004-05 Prospective European Surveillance Initiative. AB - OBJECTIVES: Telavancin is a novel semi-synthetic lipoglycopeptide currently in late-stage clinical development for the treatment of serious infections due to Gram-positive bacteria. The objective of this study was to provide a baseline prospective assessment of its in vitro activity against a large and diverse collection of Gram-positive clinical isolates from Europe and Israel. METHODS: Gram-positive clinical isolates, collected between October 2004 and December 2005 from 36 hospital laboratories in 15 countries, were tested by broth microdilution using CLSI methodology. RESULTS: In total, 3206 isolates were collected. Telavancin had potent activity against Staphylococcus aureus and coagulase negative staphylococci (MIC range < or =0.015 to 2 mg/L), independent of resistance to methicillin or to multiple drugs. Telavancin had particularly strong activity against streptococcal isolates (MIC range < or =0.001 to 0.5 mg/L), including penicillin-resistant and multiple drug-resistant Streptococcus pneumoniae and erythromycin non-susceptible beta-haemolytic and viridans group streptococci. Telavancin also had excellent activity against vancomycin susceptible enterococci (MIC(90) 0.5 mg/L), and although its MICs were elevated against VanA strains (Enterococcus faecalis MIC(90) 8 mg/L and Enterococcus faecium MIC(90) 4 mg/L), its MIC(90) was substantially lower than observed with available glycopeptides. CONCLUSIONS: Telavancin has potent in vitro activity against contemporary Gram-positive clinical isolates from diverse geographic areas in Europe and Israel. PMID- 18424793 TI - Health insurance reform in Vietnam: a review of recent developments and future challenges. AB - Vietnam is undertaking health financing reform with a view to achieve universal coverage of health insurance within the coming years. To date, around half of the population is covered with some type of health insurance or prepayment. This review applies a conceptual framework of health financing to provide a coherent assessment of the reforms to date with respect to a set of key policy objectives of health financing, including financial sustainability, efficiency in service provision, and equity in health financing. Based on the assessment, the review discusses the main implications of the reforms focusing on achievements and remaining challenges, the nature of the Vietnamese reforms in an international perspective, and the role of the government. The main lessons from the Vietnamese experiences, from which other reforming countries may draw, are the need for sustained resource mobilization, comprehensive reform involving all functions of the health financing system, and to adopt a long-term view of health insurance reform. Future analysis should include continued evaluation of the reforms in terms of impacts on key outcomes and the political dimensions of health reform. PMID- 18424794 TI - UniPrime: a workflow-based platform for improved universal primer design. AB - UniPrime is an open-source software (http://uniprime.batlab.eu), which automatically designs large sets of universal primers by simply inputting a gene ID reference. UniPrime automatically retrieves and aligns homologous sequences from GenBank, identifies regions of conservation within the alignment and generates suitable primers that can amplify variable genomic regions. UniPrime differs from previous automatic primer design programs in that all steps of primer design are automated, saved and are phylogenetically limited. We have experimentally verified the efficiency and success of this program by amplifying and sequencing four diverse genes (AOF2, EFEMP1, LRP6 and OAZ1) across multiple Orders of mammals. UniPrime is an experimentally validated, fully automated program that generates successful cross-species primers that take into account the biological aspects of the PCR. PMID- 18424795 TI - The Vienna RNA websuite. AB - The Vienna RNA Websuite is a comprehensive collection of tools for folding, design and analysis of RNA sequences. It provides a web interface to the most commonly used programs of the Vienna RNA package. Among them, we find folding of single and aligned sequences, prediction of RNA-RNA interactions, and design of sequences with a given structure. Additionally, we provide analysis of folding landscapes using the barriers program and structural RNA alignments using LocARNA. The web server together with software packages for download is freely accessible at http://rna.tbi.univie.ac.at/. PMID- 18424797 TI - Phylogeny.fr: robust phylogenetic analysis for the non-specialist. AB - Phylogenetic analyses are central to many research areas in biology and typically involve the identification of homologous sequences, their multiple alignment, the phylogenetic reconstruction and the graphical representation of the inferred tree. The Phylogeny.fr platform transparently chains programs to automatically perform these tasks. It is primarily designed for biologists with no experience in phylogeny, but can also meet the needs of specialists; the first ones will find up-to-date tools chained in a phylogeny pipeline to analyze their data in a simple and robust way, while the specialists will be able to easily build and run sophisticated analyses. Phylogeny.fr offers three main modes. The 'One Click' mode targets non-specialists and provides a ready-to-use pipeline chaining programs with recognized accuracy and speed: MUSCLE for multiple alignment, PhyML for tree building, and TreeDyn for tree rendering. All parameters are set up to suit most studies, and users only have to provide their input sequences to obtain a ready-to-print tree. The 'Advanced' mode uses the same pipeline but allows the parameters of each program to be customized by users. The 'A la Carte' mode offers more flexibility and sophistication, as users can build their own pipeline by selecting and setting up the required steps from a large choice of tools to suit their specific needs. Prior to phylogenetic analysis, users can also collect neighbors of a query sequence by running BLAST on general or specialized databases. A guide tree then helps to select neighbor sequences to be used as input for the phylogeny pipeline. Phylogeny.fr is available at: http://www.phylogeny.fr/ PMID- 18424796 TI - FireDock: a web server for fast interaction refinement in molecular docking. AB - Structural details of protein-protein interactions are invaluable for understanding and deciphering biological mechanisms. Computational docking methods aim to predict the structure of a protein-protein complex given the structures of its single components. Protein flexibility and the absence of robust scoring functions pose a great challenge in the docking field. Due to these difficulties most of the docking methods involve a two-tier approach: coarse global search for feasible orientations that treats proteins as rigid bodies, followed by an accurate refinement stage that aims to introduce flexibility into the process. The FireDock web server, presented here, is the first web server for flexible refinement and scoring of protein-protein docking solutions. It includes optimization of side-chain conformations and rigid-body orientation and allows a high-throughput refinement. The server provides a user friendly interface and a 3D visualization of the results. A docking protocol consisting of a global search by PatchDock and a refinement by FireDock was extensively tested. The protocol was successful in refining and scoring docking solution candidates for cases taken from docking benchmarks. We provide an option for using this protocol by automatic redirection of PatchDock candidate solutions to the FireDock web server for refinement. The FireDock web server is available at http://bioinfo3d.cs.tau.ac.il/FireDock/. PMID- 18424799 TI - Rapid one-step recombinational cloning. AB - As an increasing number of genes and open reading frames of unknown function are discovered, expression of the encoded proteins is critical toward establishing function. Accordingly, there is an increased need for highly efficient, high fidelity methods for directional cloning. Among the available methods, site specific recombination-based cloning techniques, which eliminate the use of restriction endonucleases and ligase, have been widely used for high-throughput (HTP) procedures. We have developed a recombination cloning method, which uses truncated recombination sites to clone PCR products directly into destination/expression vectors, thereby bypassing the requirement for first producing an entry clone. Cloning efficiencies in excess of 80% are obtained providing a highly efficient method for directional HTP cloning. PMID- 18424798 TI - Crystal structures of I-SceI complexed to nicked DNA substrates: snapshots of intermediates along the DNA cleavage reaction pathway. AB - I-SceI is a homing endonuclease that specifically cleaves an 18-bp double stranded DNA. I-SceI exhibits a strong preference for cleaving the bottom strand DNA. The published structure of I-SceI bound to an uncleaved DNA substrate provided a mechanism for bottom strand cleavage but not for top strand cleavage. To more fully elucidate the I-SceI catalytic mechanism, we determined the X-ray structures of I-SceI in complex with DNA substrates that are nicked in either the top or bottom strands. The structures resemble intermediates along the DNA cleavage reaction. In a structure containing a nick in the top strand, the spatial arrangement of metal ions is similar to that observed in the structure that contains uncleaved DNA, suggesting that cleavage of the bottom strand occurs by a common mechanism regardless of whether this strand is cleaved first or second. In the structure containing a nick in the bottom strand, a new metal binding site is present in the active site that cleaves the top strand. This new metal and a candidate nucleophilic water molecule are correctly positioned to cleave the top strand following bottom strand cleavage, providing a plausible mechanism for top strand cleavage. PMID- 18424800 TI - PharmaGist: a webserver for ligand-based pharmacophore detection. AB - Predicting molecular interactions is a major goal in rational drug design. Pharmacophore, which is the spatial arrangement of features that is essential for a molecule to interact with a specific target receptor, is an important model for achieving this goal. We present a freely available web server, named PharmaGist, for pharmacophore detection. The employed method is ligand based. Namely, it does not require the structure of the target receptor. Instead, the input is a set of structures of drug-like molecules that are known to bind to the receptor. The output consists of candidate pharmacophores that are computed by multiple flexible alignment of the input ligands. The method handles the flexibility of the input ligands explicitly and in deterministic manner within the alignment process. PharmaGist is also highly efficient, where a typical run with up to 32 drug-like molecules takes seconds to a few minutes on a stardard PC. Another important characteristic is the capability of detecting pharmacophores shared by different subsets of input molecules. This capability is a key advantage when the ligands belong to different binding modes or when the input contains outliers. The webserver has a user-friendly interface available at http://bioinfo3d.cs.tau.ac.il/PharmaGist. PMID- 18424801 TI - Prediction of phosphotyrosine signaling networks using a scoring matrix-assisted ligand identification approach. AB - Systematic identification of binding partners for modular domains such as Src homology 2 (SH2) is important for understanding the biological function of the corresponding SH2 proteins. We have developed a worldwide web-accessible computer program dubbed SMALI for scoring matrix-assisted ligand identification for SH2 domains and other signaling modules. The current version of SMALI harbors 76 unique scoring matrices for SH2 domains derived from screening oriented peptide array libraries. These scoring matrices are used to search a protein database for short peptides preferred by an SH2 domain. An experimentally determined cut-off value is used to normalize an SMALI score, therefore allowing for direct comparison in peptide-binding potential for different SH2 domains. SMALI employs distinct scoring matrices from Scansite, a popular motif-scanning program. Moreover, SMALI contains built-in filters for phosphoproteins, Gene Ontology (GO) correlation and colocalization of subject and query proteins. Compared to Scansite, SMALI exhibited improved accuracy in identifying binding peptides for SH2 domains. Applying SMALI to a group of SH2 domains identified hundreds of interactions that overlap significantly with known networks mediated by the corresponding SH2 proteins, suggesting SMALI is a useful tool for facile identification of signaling networks mediated by modular domains that recognize short linear peptide motifs. PMID- 18424803 TI - Lipophilic beta-adrenoceptor antagonist propranolol increases the hypnotic and anti-nociceptive effects of isoflurane in a swine model. AB - BACKGROUND: We have previously reported that landiolol, an ultra-short-acting beta1-adrenoceptor antagonist, does not alter the anaesthetic effects of isoflurane. Here, we investigated the influence of propranolol on the electroencephalographic (EEG) effects and minimum alveolar concentration (MAC) of isoflurane. METHODS: Fourteen swine [25.0 (SD 4.0) kg] were anaesthetized by isoflurane inhalation. The inhalation concentration was decreased to 0.5% and maintained for 25 min, before being returned to 2%, and maintained for a further 25 min. End-tidal isoflurane concentrations and spectral edge frequencies were recorded. Pharmacodynamic analysis was performed using a sigmoidal inhibitory maximal effect model for spectral edge frequency vs effect-site concentration. After measurement of the EEG effect, MAC was determined using the dew-claw clamp technique, in which movement in response to clamping is recorded. After completion of control measurements, a propranolol 4 mg bolus followed by an infusion (2 mg h(-1)) was started. After a 30 min stabilization period, the inhalation concentration of isoflurane was varied as in the control period and MAC was re-assessed. RESULTS: Propranolol shifted the concentration-effect relationship to the left and decreased the effect-site concentration that produced 50% of the maximal effect from 1.30 (0.18) to 1.13 (0.17)%. Propranolol also decreased isoflurane MAC from 1.91 (0.35) to 1.54 (0.32)%. CONCLUSIONS: Propranolol alters both the hypnotic and anti-nociceptive effects of isoflurane. In contrast to landiolol, lipophilic beta-adrenoceptor antagonists may increase the potency of inhalational anaesthetics. PMID- 18424804 TI - Analgesia from a veterinary perspective. AB - The last decade has seen continued progress in both the recognition and management of animal pain. This upsurge in the use of analgesics in animals is welcome, but the main areas of use continue to be the control of postoperative or post-trauma pain, and the management of musculoskeletal pain, in companion animals and horses. The management of pain associated with other conditions, such as soft-tissue inflammation or cancer, is still relatively neglected. Pain management in farm animals, and in animals used in biomedical research could also be improved further. Apart from providing some interesting parallels with pain management in people, development of veterinary pain management has potentially much greater significance. For many years, animal pain management has benefited from the use of analgesics used in man. In the future, it may be that a better understanding of animal pain, and in particular chronic pain states, may lead to translation of therapies in the opposite direction. PMID- 18424802 TI - Conserved residues in the delta subunit help the E. coli clamp loader, gamma complex, target primer-template DNA for clamp assembly. AB - The Escherichia coli clamp loader, gamma complex (gamma(3)deltadelta'lambdapsi), catalyzes ATP-driven assembly of beta clamps onto primer-template DNA (p/tDNA), enabling processive replication. The mechanism by which gamma complex targets p/tDNA for clamp assembly is not resolved. According to previous studies, charged/polar amino acids inside the clamp loader chamber interact with the double-stranded (ds) portion of p/tDNA. We find that dsDNA, not ssDNA, can trigger a burst of ATP hydrolysis by gamma complex and clamp assembly, but only at far higher concentrations than p/tDNA. Thus, contact between gamma complex and dsDNA is necessary and sufficient, but not optimal, for the reaction, and additional contacts with p/tDNA likely facilitate its selection as the optimal substrate for clamp assembly. We investigated whether a conserved sequence HRVW(279)QNRR--in delta subunit contributes to such interactions, since Tryptophan-279 specifically cross-links to the primer-template junction. Mutation of delta-W279 weakens gamma complex binding to p/tDNA, hampering its ability to load clamps and promote proccessive DNA replication, and additional mutations in the sequence (delta-R277, delta-R283) worsen the interaction. These data reveal a novel location in the C-terminal domain of the E. coli clamp loader that contributes to DNA binding and helps define p/tDNA as the preferred substrate for the reaction. PMID- 18424805 TI - Activated protein C inhibits chemotaxis and interleukin-6 release by human neutrophils without affecting other neutrophil functions. AB - BACKGROUND: Activated protein C (APC) therapy reduces mortality in high-risk patients with severe sepsis. The effects of APC on inflammatory responses have also been reported. Neutrophils are key cells involved in early host defence mechanisms in sepsis. We hypothesized that APC may have effects on neutrophil function. METHODS: Neutrophils were isolated from 10 healthy volunteers and incubated in the presence of lipopolysaccharide (LPS) with and without a range of therapeutically relevant concentrations of recombinant human APC. Respiratory burst activity was determined using flow-activated cell sorting (FACS) analysis. Apoptosis was determined using Annexin-V staining and FACS analysis. Cytokine bead array was used to simultaneously measure three key cytokines in culture supernatants: interleukin (IL)-1 beta, -6, and -8. For chemotaxis, neutrophil migration through a 5 microm membrane was measured in response to formyl-methyl leucine-phenylalanine (FMLP) or IL-8 in the presence and absence of APC. RESULTS: Exposure to LPS resulted in significant increases in respiratory burst activity, IL-1 beta, -6, and -8 expression (all P<0.0001) and decreased the number of apoptotic cells (P<0.0001). The APC exposure resulted in a significant release of IL-6 (P=0.04) without affecting other cytokines. Respiratory burst and apoptosis were also unaffected by APC. Neutrophil chemotaxis in response to either FMLP or IL-8 was reduced by APC (P=0.005 and 0.007, respectively). CONCLUSIONS: This pilot study showed that APC treatment of human neutrophils results in a decreased IL-6 expression and chemotaxis, without affecting other cytokines, apoptosis, or respiratory burst activity. PMID- 18424806 TI - Complications of awake fibreoptic intubation without sedation in 200 healthy anaesthetists attending a training course. AB - BACKGROUND: Two hundred anaesthetists underwent airway endoscopy and attempted awake fibreoptic intubation (FOI) on a training course. Complications were recorded and each subject's response to the procedure was assessed. METHODS: Topical airway local anaesthesia was produced with up to 9 mg kg(-1) of lidocaine, sedation was not used. Complications during and after the procedure were noted. Later, the subjects completed an anonymous questionnaire about anxiety, pain, coughing, and side-effects of lidocaine. RESULTS: More than 1300 endoscopies were performed, 180 delegates were intubated, 175 by the nasal route and five orally. Intubation was abandoned in 20 (10%) subjects. Nasal bleeding occurred in 20 (10%) subjects. Symptoms that could be attributed to lidocaine were reported by 71 (36%) subjects. Afterwards, two (1%) subjects experienced rigors and one developed a lower respiratory tract infection. CONCLUSIONS: Nasendoscopy and FOI under local anaesthesia are associated with complications, notably those of infection and airway trauma. Side-effects potentially attributable to lidocaine administration were commonly reported. PMID- 18424807 TI - Evidence for mitochondrial localization of P5, a member of the protein disulphide isomerase family. AB - This report demonstrates for the first time that P5, a member of the protein disulphide isomerase (PDI) family, is present in the mitochondria. Various organelles were screened for proteins bearing the CGHC motif using an affinity column conjugated with the phage antibody 5E, which cross-reacts with PDI family proteins. P5 was found in bovine liver mitochondrial extract and identified by Western blot analysis using anti-P5 antibody and by mass spectrometric analysis. Results of cell fractionation, proteinase sensitivity experiments and immuno electron microscopy supported the mitochondrial localization of P5 and also indicated the presence of ERp57, another PDI family protein, in mitochondria. Our findings will be useful for the elucidation of the translocation mechanism of PDI family proteins and their roles in mitochondria. PMID- 18424808 TI - Susceptibilities of phospholipid membranes containing cholesterol or ergosterol to gramicidin and its derivative incorporated in lysophospholipid micelles. AB - Complex formation of gramicidin (GA) and desformylgramicidin (des-GA) with sterols was investigated by measuring the intrinsic Trp fluorescence. In organic solvents, the Trp fluorescence of momeric GA was quenched upon binding either cholesterol or ergosterol, but that of monomeric des-GA was not quenched by adding cholesterol. Both dimeric GA and des-GA bound highly to ergosterol, but not to cholesterol, determined by quenching of Trp fluorescence. Furthermore, GA- and des-GA-loaded lysophosphatidylcholine micelles were incubated with phosphatidylcholine vesicles containing cholesterol or ergosterol. The results showed that both monomeric and dimeric peptides hardly bound to cholesterol incorporated into phospholipid vesicles, but markedly bound to ergosterol incorporated into the bilayer membranes. Interestingly, des-GA bound more specifically to the two sterols than GA. In addition, fluorescence resonance energy transfer analysis showed that des-GA bound more specifically to the two sterol than GA. PMID- 18424809 TI - Elongation and desaturation of fatty acids are critical in growth, lipid metabolism and ontogeny of Caenorhabditis elegans. AB - Recently, it was reported that a deficit in the mouse stearoyl-CoA desaturase 1 gene decreases biosynthesis and accumulation of fatty acid and revitalizes the beta-oxidation of fatty acid. To examine the physiological role of fatty acid desaturase (FAT) and elongase (ELO)-gene transduction in ontogeny, fatty acid accumulation and individual lifespan, we performed bacteria-mediated RNA interference (RNAi) in the nematode Caenorhabditis elegans. Suppression of the expression of FAT-2 gene mRNA caused a drastic decrease in the amount of body fat and defects in egg-hatching. The amount of body fat was markedly decreased, and body size reduced, by down regulation of FAT-6 and FAT-7, whereas lifespan was drastically reduced. RNAi of the FAT-2 gene caused a remarkable increase of the beta-oxidation-related gene expression and the DAF-16 transcriptional activity, whereas, ELO-2 RNAi caused a remarkable decrease in fatty acid biosynthesis related gene expression. Additionally, RNAi of FAT-6 decreased the mRNA levels of the genes involved in fatty acid synthesis, and FAT-7 RNAi increased the mRNA levels of beta-oxidation system genes. These results indicated that the elongation and desaturation of fatty acids are integral to various phenomena such as ontogeny and lifespan and play important roles in fatty acid accumulation and consumption. PMID- 18424810 TI - Modulation of glucose uptake in a human choriocarcinoma cell line (BeWo) by dietary bioactive compounds and drugs of abuse. AB - The aim of this work was to investigate the putative modulation of glucose uptake in trophoblast cells by several dietary compounds and by drugs of abuse. For this, the acute (26 min) and chronic (48 h) effect of these substances on the apical uptake of 3H-2-deoxy-D-glucose (3H-DG) by a human choriocarcinoma cell line (BeWo) was determined. 3H-DG apical uptake by BeWo cells was time dependent, displayed saturable kinetics (Vmax=1210+/-29 nmol mg protein(-1) 6 min(-1) and Km=13.4+/-0.5 mM) and was insulin-insensitive and cytochalasin B-sensitive (by up to 60%). Acutely, acetaldehyde (30-100 mM), resveratrol, xanthohumol, epigallocatechin-3-gallate (100 microM), chrysin and quercetin (10-100 microM) decreased 3H-DG apical uptake, whereas rutin, catechin (10-100 microM), epicatechin (100 microM) and ethanol (10 mM) increased it. Quercetin and xanthohumol seem to be non-competitive inhibitors of 3H-DG apical uptake, whereas both epigallocatechin-3-gallate and acetaldehyde decreased both the Km and Vmax values. Chronically, rutin and myricetin increased the apical uptake of 3H-DG both isolated (0.1-1 microM) and in combination (both at 1 microM), whereas theophylline (0.1-1 microM) and amphetamine, 3,4-methylenedioxymethamphetamine (0.25-1 microM) and Delta9-tetrahydrocannabinol (1 nM) decreased it. In conclusion, 3H-DG apical uptake by BeWo cells is differentially modulated by different compounds present in drinks and by drugs of abuse. PMID- 18424811 TI - 15N NMR relaxation studies of Y14F mutant of ketosteroid isomerase: the influence of mutation on backbone mobility. AB - The backbone dynamics of Y14F mutant of Delta(5)-3-ketosteroid isomerase (KSI) from Comamonas testosteroni has been studied in free enzyme and its complex with a steroid analogue, 19-nortestosterone hemisuccinate (19-NTHS), by 15N NMR relaxation measurements. Model-free analysis of the relaxation data showed that the single-point mutation induced a substantial decrease in the order parameters (S2) in free Y14F KSI, indicating that the backbone structures of Y14F KSI became significantly mobile by mutation, while the chemical shift analysis indicated that the structural perturbations of Y14F KSI were more profound than those of wild-type (WT) KSI upon 19-NTHS binding. In the 19-NTHS complexed Y14F KSI, however, the key active site residues including Tyr14, Asp38 and Asp99 or the regions around them remained flexible with significantly reduced S2 values, whereas the S2 values for many of the residues in Y14F KSI became even greater than those of WT KSI upon 19-NTHS binding. The results thus suggest that the hydrogen bond network in the active site might be disrupted by the Y14F mutation, resulting in a loss of the direct interactions between the catalytic residues and 19-NTHS. PMID- 18424812 TI - Humanization of an anti-CD34 monoclonal antibody by complementarity-determining region grafting based on computer-assisted molecular modelling. AB - 4C8 is a new mouse anti-human CD34 monoclonal antibody (mAb), which recognizes class II CD34 epitopes and can be used for clinical hematopoietic stem/progenitor cell selection. In an attempt to improve its safety profiles, we have developed a humanized antibody of 4C8 by complementarity-determining region (CDR) grafting method in this study. Using a molecular model of 4C8 built by computer-assisted homology modelling, framework region (FR) residues of potential importance to the antigen binding were identified. A humanized version of 4C8, denoted as h4C8, was generated by transferring these key murine FR residues onto a human antibody framework that was selected based on homology to the mouse antibody framework, together with the mouse CDR residues. The resultant humanized antibody was shown to possess antigen-binding affinity and specificity similar to that of the original murine antibody, suggesting that it might be an alternative to mouse anti-CD34 antibodies routinely used clinically. PMID- 18424813 TI - Improving the scale and precision of hypotheses to explain root foraging ability. AB - BACKGROUND: Numerous hypotheses have been proposed to explain the wide variation in the ability of plants to forage for resources by proliferating roots in soil nutrient patches. Comparative analyses have found little evidence to support many of these hypotheses, raising the question of what role resource-foraging ability plays in determining plant fitness and community structure. SCOPE: In the present viewpoint, we respond to Grime's (2007; Annals of Botany 99: 1017-1021) suggestion that we misinterpreted the scope of the scale-precision trade-off hypothesis, which states that there is a trade-off between the spatial scale over which plant species forage and the precision with which they are able to proliferate roots in resource patches. We use a meta-analysis of published foraging scale-precision correlations to demonstrate that there is no empirical support for the scale-precision trade-off hypothesis. Based on correlations between foraging precision and various plant morphological and ecophysiological traits, we found that foraging precision forms part of the 'fast' suite of plant traits related to rapid growth rates and resource uptake rates. CONCLUSIONS: We suggest there is a need not only to examine correlations between foraging precision and other plant traits, but to expand our notion of what traits might be important in determining the resource-foraging ability of plants. By placing foraging ability in the broader context of plant traits and resource economy strategies, it will be possible to develop a new and empirically supported framework to understand how plasticity in resource uptake and allocation affect plant fitness and community structure. PMID- 18424814 TI - Preoperative diagnosis of lymph node metastases of colorectal cancer by FDG PET/CT. AB - PURPOSE: The purpose of this study was to assess the diagnostic value of 18F fluorodeoxyglucose positron emission tomography/computed tomography (FDG-PET/CT) for lymph node (LN) metastasis of colorectal cancer. METHODS: FDG-PET/CT was used to preoperatively evaluate 88 patients with colorectal cancer. In this study, LN sites were divided into proximal and distant according to their distance from the primary tumor. The FDG-PET/CT images were evaluated by three criteria; nodal diameter, abnormal uptake and maximum standardized uptake value (SUV). We compared the diagnostic ability of these methods for LN metastasis at proximal and distant sites. RESULTS: The mean SUV of the malignant LNs was significantly higher than that of the benign LNs. The sensitivity, specificity and accuracy of diagnosis by abnormal uptake were 28.6, 92.9 and 75.0%, those by nodal diameter using cutoff value of 10 mm were 30.6, 95.3 and 74.4% and those by SUV using cutoff value of 1.5 were 53.1, 90.6 and 80.1%, respectively. The sensitivity, specificity and accuracy of diagnosis based on optimal SUV were 51.2, 85.1 and 69.3% in the proximal site and 62.5, 92.5 and 89.7%, respectively, in the distant site. CONCLUSIONS: FDG-PET/CT is useful for preoperative diagnosis of distant LN metastases of colorectal cancers. PMID- 18424815 TI - A note on the false discovery rate and inconsistent comparisons between experiments. AB - MOTIVATION: The false discovery rate (FDR) has been widely adopted to address the multiple comparisons issue in high-throughput experiments such as microarray gene expression studies. However, while the FDR is quite useful as an approach to limit false discoveries within a single experiment, like other multiple comparison corrections it may be an inappropriate way to compare results across experiments. This article uses several examples based on gene-expression data to demonstrate the potential misinterpretations that can arise from using FDR to compare across experiments. Researchers should be aware of these pitfalls and wary of using FDR to compare experimental results. FDR should be augmented with other measures such as p-values and expression ratios. It is worth including standard error and variance information for meta-analyses and, if possible, the raw data for re-analyses. This is especially important for high-throughput studies because data are often re-used for different objectives, including comparing common elements across many experiments. No single error rate or data summary may be appropriate for all of the different objectives. PMID- 18424816 TI - IMGT, a system and an ontology that bridge biological and computational spheres in bioinformatics. AB - IMGT, the international ImMunoGeneTics information system (http://imgt.cines.fr), is the reference in immunogenetics and immunoinformatics. IMGT standardizes and manages the complex immunogenetic data that include the immunoglobulins (IG) or antibodies, the T cell receptors (TR), the major histocompatibility complex (MHC) and the related proteins of the immune system (RPI), which belong to the immunoglobulin superfamily (IgSF) and the MHC superfamily (MhcSF). The accuracy and consistency of IMGT data and the coherence between the different IMGT components (databases, tools and Web resources) are based on IMGT-ONTOLOGY, the first ontology for immunogenetics and immunoinformatics. IMGT-ONTOLOGY manages the immunogenetics knowledge through diverse facets relying on seven axioms, 'IDENTIFICATION', 'DESCRIPTION', 'CLASSIFICATION', 'NUMEROTATION', 'LOCALIZATION', 'ORIENTATION' and 'OBTENTION', that postulate that objects, processes and relations have to be identified, described, classified, numerotated, localized, orientated, and that the way they are obtained has to be determined. These axioms constitute the Formal IMGT-ONTOLOGY, also designated as IMGT-Kaleidoscope. These axioms have been essential for the conceptualization of the molecular immunogenetics knowledge and for the creation of IMGT. Indeed all the components of the IMGT integrated system have been developed, based on standardized concepts and relations, thus allowing IMGT to bridge biological and computational spheres in bioinformatics. The same axioms can be used to generate concepts for multi-scale level approaches at the molecule, cell, tissue, organ, organism or population level, emphasizing the generalization of the application domain. In that way the Formal IMGT-ONTOLOGY represents a paradigm for the elaboration of ontologies in system biology. PMID- 18424817 TI - Is rapid initiation of peritoneal dialysis feasible in unplanned dialysis patients? A single-centre experience. AB - BACKGROUND: Starting dialysis in an unplanned manner is a frequent situation in a dialysis centre even for patients with a regular nephrology follow-up. For no clear reason, chronic haemodialysis (HD) is more frequently used than peritoneal dialysis for unplanned dialysis patients. OBJECTIVE: The purpose of this study was to evaluate the results of a dialysis policy dedicated to unplanned dialysis patients. The aim of this policy was to increase the use of peritoneal dialysis (PD) in an attempt to reduce the need for tunnelled catheter. METHODS: One hundred seventy-one patients from a single centre, who started dialysis between 1 January 2004 and 31 December 2006, were prospectively followed until 31 December 2006. Unplanned dialysis patients were defined as patients entering in dialysis with no vascular access or peritoneal dialysis catheter. PD was presented as a modality of choice for renal replacement therapy to avoid the need for a tunnelled HD catheter. RESULTS: There were 60 unplanned dialysis patients during the study period. Among these patients, 34 agreed to be treated by PD. Compared with unplanned peritoneal dialysis patients, unplanned haemodialysis patients had a greater modified Charlson's comorbidity index (5.9 +/- 2.4 versus 4.4 +/- 1.9, P < 0.05). The mean duration of the temporary catheter period was 32 +/- 29 days (median: 24 days) for haemodialysis patients compared with 26 +/- 21 days (median: 25 days) for peritoneal dialysis patients (P = NS). The initial hospitalization duration was similar in haemodialysis patients and peritoneal dialysis patients (24 +/- 28 versus 30 +/- 33 days; median value: 17 versus 20 days, P = NS). PD was started 8.6 +/- 10 days (median: 4 days) after catheter insertion. A tunnelled catheter was used only in three patients until peritoneal dialysis was initiated. Acute automated peritoneal dialysis was used in 19 patients. Among 26 haemodialysis patients, 23 were dialyzed through a tunnelled catheter. Of these 23 patients, 15 were successfully converted to fistula. Median time for fistula creation was 2.6 months after dialysis initiation; median time for fistula utilization was 4.4 months. Actuarial patients survival at 1 year was 79% on haemodialysis compared with 83% on peritoneal dialysis (P = NS). After adjustment of the initial modified Charlson's comorbidity index, dialysis modality had no impact on patient's survival. There was no significant difference between haemodialysis patients and peritoneal dialysis patients regarding survival free of re-hospitalization. Actuarial survival free of peritonitis was 73% at 6 months and 58% at 1 year. CONCLUSION: Peritoneal dialysis is a safe and efficient alternative to haemodialysis for unplanned dialysis patients. Peritoneal dialysis offers the advantage of reducing the need for tunnelled catheter in unplanned dialysis patients. PMID- 18424818 TI - Proteomic profiling and identification in peritoneal fluid of children treated by peritoneal dialysis. AB - BACKGROUND: Proteomic technologies offer a high-throughput analysis of the expression of proteins in biological samples. The global analysis of the proteins in peritoneal dialysis (PD) fluid will provide a better understanding of the biological processes of the peritoneal membrane. METHODS: The dialysate of nine paediatric PD patients was collected from peritoneal equilibrium tests with 3.86% glucose. Proteins were separated on a 10% SDS-PAGE gel and in-gel digested with trypsin. Peptide mixtures were analysed using nanoLC-MS/MS and results were searched against the NCBI database. RESULTS: A total number of 189 proteins were identified in the PD fluid of nine patients, with 88 proteins shared by all patients. These 88 proteins accounted for 47% of the identified proteins and >90% of the total protein content in the analysed samples. Proteins were subdivided into eight different classes according to function. CONCLUSIONS: This study gives a representative overview of the proteins present in PD fluid. The proteins in PD fluid reflect plasma proteins as well as local peritoneal processes. Potentially interesting proteins are revealed. PMID- 18424819 TI - Renal transplantation in adults with Henoch-Schonlein purpura: long-term outcome. AB - BACKGROUND: Little information is available about the long-term outcome of renal transplantation in adults with Henoch-Schonlein purpura (HSP). METHODS: We compared the outcomes of 17 patients with HSP who received 19 renal transplants with those of 38 controls matched for time of transplantation, age, gender and source of donor. The mean post-transplant follow-up was 109 +/- 99 months for HSP patients and 110 +/- 78 months for controls. RESULTS: The actuarial 15-year patient survival was 80% in HSP patients and 82% in controls, and the death censored graft survival was 64% in HSP patients and in controls. The risks of acute rejection, chronic graft dysfunction, arterial hypertension and infection were not different between the two groups. In eight grafts (42%) recurrence of HSP nephritis was found (0.05/patient/year). In spite of therapy, one patient died and four eventually restarted dialysis respectively 10, 32, 35 and 143 months after renal transplant. Seventy-one percent of grafts transplanted in patients with necrotizing/crescentic glomerulonephritis of the native kidney had HSP recurrence in comparison to 12% of recurrences in patients with mesangial nephritis (P = 0.05) CONCLUSIONS: Long-term patient and allograft survival of HSP patients was good. However, 42% of HSP patients, particularly those with necrotizing/crescentic glomerulonephritis of the native kidneys, developed a recurrence of HSP nephritis that eventually caused the loss of the graft function in half of them. PMID- 18424820 TI - Calcium-mediated parathyroid hormone release changes in patients treated with the calcimimetic agent cinacalcet. AB - BACKGROUND: The parathyroid-calcium (Ca(2+)-PTH) curve expresses modulation of parathyroid hormone (PTH) secretion by the parathyroid gland as a function of changing extracellular Ca(2+) concentration. Patients with hyperparathyroidism (HPT) show a rightward shift of the curve compared with controls, suggesting a reduced sensitivity of parathyroid cells to Ca(2+). Increasing the sensitivity of the parathyroid gland to extracellular Ca(2+) by manipulation of the Ca(2+) sensing receptor (CaR) may have therapeutic potential. Calcimimetics allosterically modify CaR and render it more sensitive to extracellular Ca(2+), accounting for the simultaneous reduction of Ca(2+) and PTH seen in most patients. METHODS: The Ca(2+)-PTH curve was evaluated in 10 haemodialysis patients, with baseline intact PTH levels >300 pg/ml in two haemodialysis sessions, one before and the other after (range, 9-22 weeks) cinacalcet treatment. In each session a 2-h low-dialysate Ca(2+) concentration was used to induce hypocalcaemia and maximally stimulate PTH secretion, followed immediately by a 2-h high-dialysate Ca(2+) concentration to induce hypercalcaemia and maximally inhibit PTH secretion. RESULTS: Significant decreases in ionized Ca(2+) and intact PTH were observed following cinacalcet treatment. Cinacalcet treatment also led to a decrease in the set point for Ca(2+) and to a leftward shift of the Ca(2+)-PTH curve. Significant differences were present in all segments of the Ca(2+)-PTH curves. CONCLUSION: The pathological rightward shift of the Ca(2+)-PTH curve seen in many HPT patients may be reversed by cinacalcet treatment. PMID- 18424821 TI - Improving activity and stability of cutinase towards the anionic detergent AOT by complete saturation mutagenesis. AB - Cutinase is an enzyme suitable for detergent applications as well as for organic synthesis in non-aqueous solvents. However, its inactivation in the presence of anionic surfactants is a problem which we have addressed by creating a complete saturation library. For this, the cutinase gene from Fusarium solani pisi was mutated to incorporate all 19 possible amino acid exchanges at each of the 214 amino acid positions. The resulting library was screened for active variants with improved stability in the presence of the anionic surfactant dioctyl sulfosuccinate sodium salt (AOT). Twenty-four sites in cutinase were discovered where amino acid replacements resulted in a 2-11-fold stability increase as compared to the wild-type enzyme. PMID- 18424822 TI - Thomas Carlyle Jones. PMID- 18424823 TI - Should we kill the case report? PMID- 18424824 TI - Estrogens metabolism associated with polymorphisms: influence of COMT G482a genotype on age at onset of canine mammary tumors. AB - Catechol-O-methyltransferase (COMT) is an important enzyme participating in inactivation of carcinogenic oestrogen metabolites. In humans there is a single nucleotide polymorphism in COMT gene (COMT val158met) that has been associated with an increased risk for developing breast cancer. In dogs, there is a single nucleotide polymorphism in COMT gene (G482A), but its relation with mammary carcinogenesis has never been investigated. The aim of this study was to focus on the evaluation of such polymorphism as a risk factor for the development of mammary tumors in bitches and on the analysis of its relationship with some clinicopathologic features (dog's age and weight, number and histologic type of the lesions, lymph node metastasis) of canine mammary neoplasms. A case-control study was conducted analyzing 90 bitches with mammary tumors and 84 bitches without evidence of neoplastic disease. The COMT G482A polymorphism was analyzed by PCR-RFLP. We found a protective effect of the polymorphism in age of onset of mammary tumors, although we could not establish a significant association between COMT genotype and other clinicopathologic parameters nor with mammary tumor risk overall. Animals carrying the variant allele have a threefold likelihood of developing mammary tumors after 9 years of age in comparison with noncarriers. The Kaplan-Meier method revealed significant differences in the waiting time for onset of malignant disease for A allele carrier (12.46 years) and noncarrier (11.13 years) animals. This investigation constitutes the first case-control study designed to assess the relationship between polymorphic genes and mammary tumor risk in dogs. Our results point to the combined effect of COMT genotype with other genetic and/or environmental risk factors as important key factors for mammary tumor etiopathogenesis. PMID- 18424825 TI - Vascular endothelial growth factor mRNA expression and peritumoral edema in canine primary central nervous system tumors. AB - Vascular endothelial growth factor (VEGF) is an important regulator of tumor angiogenesis and vascular permeability, and has been implicated both in progression of central nervous system (CNS) tumors and development of vasogenic peritumoral edema. A retrospective study was done to characterize the levels of expression of the 3 major canine VEGF isoforms (VEGF(120), VEGF(164), VEGF(188)) in a variety of spontaneous canine CNS tumors using quantitative TaqMan reverse transcription real-time polymerase chain reaction. Presence and degree of peritumoral edema also were determined in sampled tumors using magnetic resonance imaging (MRI). Increased expression of VEGF relative to normal cerebral cortex tissue was seen predominantly in high grade astrocytic (grade IV) and oligodendroglial (grade III) tumors, with lower expression in low grade astrocytomas (grade II) and meningiomas (grade I). All 3 major VEGF isoforms were present; VEGF(164) was the predominant isoform, particularly in the tumors with the highest VEGF expression. Peritumoral edema was present in all tumor types; however, a significant association between the extent of peritumoral edema and the level of VEGF expression was not apparent. PMID- 18424827 TI - Polysaccharide storage myopathy in Cob Normand draft horses. AB - Gluteus medius muscle was sampled from 53 Cob Normand horses for histologic evaluation. Twenty horses (38%) exhibited amylase-resistant material in myocytes consistent with polysaccharide storage myopathy. Diameter of affected type II fibers was increased (67.7 +/- 21.4 microm) compared with normal ones (57.3 +/- 19.7 microm). Two groups were distinguished by quantitative study. The first group (n = 14; 26%) was characterized by a low percentage of fibers (m = 0.98%) containing aggregates occurring singly or in perifascicular clusters without myopathic changes. The second group (n = 6; 11%) was characterized by a high percentage (m = 18.1%) of fibers containing aggregates scattered in biopsy with chronic myopathic changes. Re-biopsy of 4 horses showed an increase with time in the number of aggregate-containing fibers for horses of the first group only. In 1 necropsied horse, aggregates were observed in a wide range of muscles including smooth muscles. Ultrastructurally, granular material was found interspersed among arrays of filamentous material. PMID- 18424826 TI - Detection of antigenic heterogeneity in feline coronavirus nucleocapsid in feline pyogranulomatous meningoencephalitis. AB - A new monoclonal antibody (mAb), CCV2-2, was compared with the widely used FIPV3 70 mAb, both directed against canine coronavirus (CCoV), as a diagnostic and research tool. Western blot showed that both anti-CCoV mAbs only reacted with a protein of 50 kD, a weight consistent with the feline coronavirus (FCoV) viral nucleocapsid. A competitive inhibition enzyme-linked immunosorbent assay showed that the 2 recognized epitopes are distinct. Preincubation of CCV2-2 mAb with FCoV antigen suppressed the immunostaining. Formalin-fixed, paraffin-embedded sections from brains of 15 cats with the dry form of feline infectious peritonitis (FIP) were examined by immunohistochemistry. Immunohistochemistry was performed with both anti-CCoV mAbs, either on consecutive or on the same sections. A myeloid-histiocytic marker, MAC 387, was also used to identify FIP virus-infected cells. In all regions where MAC 387-positive cells were present, positive staining with the CCV2-2 mAb was systematically detected, except at some levels in 1 cat. In contrast, none or only a few cells were positive for the FIPV3-70 mAb. Double immunostaining showed macrophages that were immunopositive for either CCV2-2 alone or alternatively for CCV2-2 and FIPV3-70 mAbs. This reveals the coexistence of 2 cohorts of phagocytes whose FIP viral contents differed by the presence or absence of the FIPV3-70-recognized epitope. These findings provide evidence for antigenic heterogeneity in coronavirus nucleocapsid protein in FIP lesions, a result that is in line with molecular observations. In addition, we provide for the first time morphologic depiction of viral variants distribution in these lesions. PMID- 18424828 TI - Histopathologic, immunohistochemical, and cytologic analysis of feline myeloma related disorders: further evidence for primary extramedullary development in the cat. AB - Feline myeloma-related disorders (MRD) are rare neoplasms of plasma cells. The multistep transformation model of myeloma in humans is based on the premise that plasma cells undergo neoplastic transformation primarily within the intramedullary compartment and that over time they become poorly differentiated and metastasize to extramedullary locations. Historically, diagnostic criteria used for human multiple myeloma have been applied to the cat, with the assumption that feline MRD commonly arises in the intramedullary compartment. Our objectives were to describe the features of feline MRD confirmed by cytology, histopathology, histochemistry, and immunohistochemistry and to categorize these tumors. A priori hypotheses were 1) tumor category predicts survival and 2) cats with well-differentiated tumors commonly have extramedullary involvement in contrast to human myeloma patients. This multicenter, retrospective study identified 26 MRD cases. There was good agreement between histopathologic and cytologic tumor categorization. Histochemistry and immunohistochemistry were shown to be valuable adjunct tests in the diagnosis of MRD. Cats with well differentiated tumors had increased median survival relative to those with poorly differentiated tumors (254 versus 14 days). We have reported that marked extramedullary involvement at initial clinical presentation is significantly more common in the cat than in human MRD patients. In this study, we demonstrate that cats with well-differentiated tumors more commonly have extramedullary involvement than human myeloma patients with well-differentiated tumors (90% versus 20%, P < 0.0002). These results contrast strongly with the human myeloma model of primary intramedullary neoplastic transformation and suggest that primary extramedullary neoplastic transformation may be more common in feline MRD. PMID- 18424830 TI - Nasal and paranasal adenocarcinomas with neuroendocrine differentiation in dogs. AB - Tumors of the nasal cavity or paranasal sinuses of 18 dogs were examined histopathologically, immunohistochemically, and histochemically. The tumors were classified histologically as 13 adenocarcinomas, 3 transitional carcinomas, 1 squamous cell carcinoma, and 1 adenosquamous carcinoma. Tumor cells were strongly immunoreactive for broad-spectrum cytokeratins in all cases, for cytokeratin 8/18 in 16 cases, and for cytokeratin 19 in 17 cases. None of the 18 carcinomas had cytologic or histologic features indicative of neuroendocrine differentiation, yet tumor cells in 5 of the 13 adenocarcinomas were argyrophilic and immunohistochemically positive for synaptophysin and chromogranin A. Results of this study indicate that neuroendocrine markers may be detected immunohistochemically and histochemically in canine nasal or paranasal adenocarcinomas despite the lack of typical histologic features of neuroendocrine differentiation. PMID- 18424829 TI - The clinical and morphologic features of nonepidermolytic ichthyosis in the golden retriever. AB - A scaling disorder specific to Golden Retriever dogs has been recognized by both dermatologists and pathologists, but to date has not been well characterized. At the University of Pennsylvania's Laboratory of Toxicology and Pathology, 46 cases of ichthyosis were diagnosed histologically in Golden Retriever dogs from January 2004 to January 2007. A total of 22 dogs had skin lesions documented at younger than 1 year of age; 3 dogs between 1 and 2 years of age; 13 dogs developed lesions at older than 2 years; and the time of onset was unknown for 8 dogs. A total of 25 dogs were female, and 21 were male. All dogs had strikingly similar histopathologic changes that consisted of mild to moderate laminar orthokeratotic hyperkeratosis with an absence of epidermal hyperplasia and dermal inflammation. Ultrastructural analysis using a ruthenium tetroxide fixation method was performed on punch biopsy samples from 5 dogs and compared with 2 control dogs (1 clinically and histologically normal sibling of an affected dog and 1 Cairn Terrier). All affected dogs had retained and convoluted membranes with crystalline structures in the stratum corneum. Scattered keratinocytes in the granular cell layer had prominent, clear, membrane-bound, cytoplasmic vacuoles. Pedigree analysis of 14 dogs was compatible with autosomal recessive inheritance, but incomplete dominance could not be ruled out. This unique hyperkeratotic/scaling disorder in Golden Retrievers has distinctive clinical, histologic, and ultrastructural features, which are consistent with a primary cornification defect. PMID- 18424831 TI - Multiple cystic intestinal duplications in a cat. AB - A 7-year-old, female European shorthair cat with a history of recurrent vomiting had a 2-cm cystic mass in the midjejunum. Cross-sectioning and histology revealed 3 separate cystic structures in the muscular layer, in addition to a regularly structured intestinal lumen. One cyst had a 3-layered wall consisting of a dysplastic mucosa, a regularly structured submucosa, and partly double-layered muscularis that sporadically contained neurons resembling a myenteric plexus. The remaining 2 cysts had similar structures except for granulation tissue lining the lumen. The lesion was diagnosed as multiple cystic duplications in the midjejunum, which is unknown to the veterinary literature to date. PMID- 18424832 TI - Concurrent peste des petits ruminants virus and pestivirus infection in stillborn twin lambs. AB - Concurrent infection with peste des petits ruminants virus (PPRV) and pestivirus was diagnosed in stillborn twin lambs. With the flock history, the findings of epidermal syncytial cells and necrotizing bronchitis/bronchiolitis prompted testing for PPRV infection, and PPRV antigen was detected by immunohistochemistry (IHC) in the skin, lungs, kidneys, rumen, and thymus. Macroscopic anomalies that were typical of border disease included scoliosis, brachygnathism, prognathism, arthrogryposis, hydranencephaly, cerebellar hypoplasia, and hairy fleece; pestiviral antigen was detected by IHC in the brain, liver, lungs, and kidneys. Tissues from both lambs were positive by reverse transcriptase-polymerase chain reaction (RT-PCR) for PPRV and pestivirus. To the authors' knowledge, PPR has not been reported previously as a congenital infection or in combination with pestiviral infection. PMID- 18424833 TI - Bone dysplasia in the radial and ulnar metaphysis of a Newfoundland dog. AB - A recently described metaphyseal irregularity of the radius and ulna was diagnosed radiographically in a significant proportion of Newfoundland dogs during the course of a large study. This case report describes the pathological picture of a Newfoundland dog with these radiographic changes. The lesions in the distal radius and ulna were characterized by focal, longitudinal striations of sclerosis of the bone marrow cavity, surrounding thin trabeculae of primary spongiosa of the distal metaphysis. It is suggested that these lesions represent a sclerosing dysplasia not previously described in dogs, but with some similarities to the human disorder, osteopathia striata. PMID- 18424834 TI - Sphingomyelinase deficiency (Niemann-Pick disease) in a Hereford calf. AB - A 5-month-old Hereford calf with neurologic disease was euthanatized, and a necropsy was done. No gross lesions were seen in the brain. Microscopically, neurons throughout the brain and spinal cord had distended, foamy vacuolated cytoplasm. Ultrastructure showed clear vacuoles filling the neuronal cytoplasm. A lysosomal storage disease was suspected. Sphingomyelinase deficiency was confirmed by biochemical analysis of liver and brain. PMID- 18424835 TI - Mandibular ossifying fibroma in a dog. AB - An aged mongrel dog was admitted for hemimandibulectomy as treatment for a mandibular mass that had been diagnosed as osteosarcoma. The fibro-osseous mass that surrounded the first molar tooth and replaced alveolar and cortical bone was reclassified as ossifying fibroma on the basis of anatomic location and histologic features. The tumor was composed of isomorphic fusiform cells with few mitotic figures. Tumoral stroma contained trabeculae of woven bone that were bordered by a single layer of osteoblasts. Excision was deemed complete with no evidence of extension or metastasis by computed tomography of the skull or thoracic and abdominal radiography. The dog was reportedly healthy 6 months after initial presentation. Though far less common than osteosarcoma as a primary canine bone tumor, ossifying fibroma should be included in the differential diagnosis for fibro-osseous proliferations, especially those of the jaw. Although benign, en bloc excision may be necessary for surgical cure. PMID- 18424836 TI - Xanthogranulomatous inflammation of the small bowel in a dog. AB - A case of xanthogranulomatous inflammation of the small bowel in a 12-year-old male American Staffordshire Terrier is described. Disseminated yellow-white nodules 2 to 3 mm in diameter bulging on the serosal surface of the small bowel, as well as on mesenteric tissue, were detected. Histopathologic examination revealed a nodular collection of foamy cells, mainly involving serosal and muscular layers, associated with necrotic areas, hemorrhages, neovascularization, variable numbers of reactive spindle cells, neutrophils, lymphocytes, plasma cells, and rare multinucleated giant cells. Transmural lymphangectasia and mucosal lymphoplasmacytic inflammation were also observed. Both Oil Red O stain and ultrastructural study revealed lipid droplets in the cytoplasm of foamy cells. Lysozyme immunoreactivity was detected in single as well as in clustered foamy cells, while smooth muscle actin was positive in spindle cells and scattered foamy elements. Lymphangectasia associated with lymphoplasmacytic enteritis suggests a component of lymphatic fluid stasis in the pathogenesis of such lesions. PMID- 18424837 TI - Herpetiform genital lesions in a heifer with mucosal disease. AB - A 14-month-old heifer with a 17-day history of unresponsive bloody diarrhea was necropsied. There were focal, pink-red erosions of the nares and hard palate; ulcers and fissures of the tongue; and multiple ulcerative lesions of the alimentary canal. Interdigital skin of both rear limbs was ulcerated and bleeding; and the margins of the vulva contained punctiform red ulcers. The gross lesions were consistent with mucosal disease. Histopathology and laboratory testing ruled out rinderpest, foot-and-mouth disease, and vesicular stomatitis, and identified bovine virus diarrhea virus to be the cause of this disease. Lesions of the vulva similar to those seen in some stages of infectious pustular vulvovaginitis were negative for bovine herpesvirus-1 and tested positive for bovine viral diarrhea virus antigen by immunohistochemistry. PMID- 18424838 TI - The immunohistochemical evaluation of estrogen receptor-alpha and progesterone receptors of normal, hyperplastic, and neoplastic endometrium in 88 pet rabbits. AB - The expression of estrogen receptor-alpha (ER) and progesterone receptor (PR) in normal, hyperplastic, and neoplastic endometrium in rabbits was evaluated by immunohistochemistry. The tissues evaluated were 27 normal uteri, 19 cases with endometrial hyperplasia, and 42 adenocarcinomas. Sixteen of 27 cases of normal uteri (59.3%) and 13 out of 19 hyperplasias (68.4%) stained positive with both ER alpha and PR. Adenocarcinomas were further subdivided into 26 papillary and 16 tubular/solid adenocarcinomas. Papillary adenocarcinoma infiltrated the myometrium late in the disease and caused attenuation of the myometrium. In contrast, tubular/solid adenocarcinoma invaded into the deep myometrium early in the disease without thinning of the myometrium. Twenty-one cases out of 26 (80.8%) cases of papillary adenocarcinoma were both ER-alpha and PR negative, whereas 15 out of 16 (93.8%) of the tubular/solid adenocarcinomas were positive for ER-alpha, PR, or both. The total immunoreactive scores of ER-alpha, PR, and mode of myometrial invasion were significantly different between histopathologic types. This suggests that there may be 2 different developmental pathways for uterine adenocarcinomas in the rabbit. PMID- 18424839 TI - Multisystemic abscesses in African green monkeys (Chlorocebus aethiops) with invasive Klebsiella pneumoniae--identification of the hypermucoviscosity phenotype. AB - Invasive Klebsiella pneumoniae is an emerging disease of humans characterized by abscesses in the liver or other sites involving bacteria with the unique hypermucoviscosity phenotype. Over several months, 7 African green monkeys in our research colony developed abscess formation in multiple locations and succumbed to disease. K. pneumoniae was identified by bacterial culture in 6 monkeys and immunohistochemistry in 1 additional monkey. All monkeys had been housed in, or had contact with monkeys housed in, 1 animal room in our facility. All affected monkeys had 1 or more abscesses, most notably in the abdomen, but also affecting the lungs, cerebellum, and skin. Abdominal abscesses and associated adhesions entrapped loops of bowel, forming palpable masses. Abdominal masses were located at the root of the mesentery, the ileocecocolic junction, or the pelvic inlet. In 1 case, culture, serotyping, and polymerase chain reaction (PCR) analysis of the bacterial isolate identified K. pneumoniae expressing the hypermucoviscosity phenotype and capsular serotype K2 and determined that the K. pneumonia was genetically rmpA(+)/magA(-). PMID- 18424840 TI - Congenital cystic adenomatoid-like malformation in a cynomolgus monkey (Macaca fascicularis). AB - Congenital cystic adenomatoid malformation (CCAM) is a developmental lung abnormality characterized by abnormal proliferation of mesenchymal elements and failure of bronchiolar structures to mature, ultimately resulting in the compression of normal pulmonary tissue and mediastinal shift with rapid expansion of cysts. Although various clinical and pathologic studies of CCAM in humans exist, CCAM has yet to be reported in animals, even in nonhuman primates. In the present study, histopathologic analyses of a neonatal cynomolgus monkey that died 17 days after birth revealed that normal lung architecture was replaced by disorganized overgrowths of cysts lined with simple cuboidal epithelium. The epithelium projected a few ciliates into the air spaces and produced mucus. To our knowledge, this is the first case study describing CCAM or a CCAM-like lesion in nonhuman primates. PMID- 18424841 TI - Clinicopathologic features of a systemic coronavirus-associated disease resembling feline infectious peritonitis in the domestic ferret (Mustela putorius). AB - From 2002 to 2007, 23 ferrets from Europe and the United States were diagnosed with systemic pyogranulomatous inflammation resembling feline infectious peritonitis (FIP). The average age at the time of diagnosis was 11 months. The disease was progressive in all cases, and average duration of clinical illness was 67 days. Common clinical findings were anorexia, weight loss, diarrhea, and large, palpable intra-abdominal masses; less frequent findings included hind limb paresis, central nervous system signs, vomiting, and dyspnea. Frequent hematologic findings were mild anemia, thrombocytopenia, and hypergammaglobulinemia. Grossly, whitish nodules were found in numerous tissues, most frequently the mesenteric adipose tissue and lymph nodes, visceral peritoneum, liver, kidneys, spleen, and lungs. One ferret had a serous abdominal effusion. Microscopically, pyogranulomatous inflammation involved especially the visceral peritoneum, mesenteric adipose tissue, liver, lungs, kidneys, lymph nodes, spleen, pancreas, adrenal glands, and/or blood vessels. Immunohistochemically, all cases were positive for coronavirus antigen using monoclonal antibody FIPV3-70. Electron microscopic examination of inflammatory lesions identified particles with coronavirus morphology in the cytoplasm of macrophages. Partial sequencing of the coronavirus spike gene obtained from frozen tissue indicates that the virus is related to ferret enteric coronavirus. PMID- 18424842 TI - Fatal coxiellosis in Swainson's Blue Mountain Rainbow Lorikeets (Trichoglossus haematodus moluccanus). AB - Three Swainson's Blue Mountain Rainbow Lorikeets (Trichoglossus haematodus moluccanus), ranging from 6 to 8 months of age, presented with lethargy, emaciation, and progressive neurologic signs. The first one died 24 hours after the onset of clinical signs, and the other two were euthanized 10 to 14 days after the onset of progressive neurologic disease. Clinical signs in these lorikeets included head pressing, hemiparesis, seizures, obtunded mentation, weakness, and lethargy. Two of the lorikeets had hepatomegaly, and one had splenomegaly on gross examination. Histopathology revealed disseminated microgranulomas in the liver, spleen, and brain, and lymphohistocytic perivascular encephalitis and cephalic vasculitis. Electron microscopic examination of macrophages in brain lesions revealed spherical to rod-shaped prokaryotic organisms with a trilaminar cell wall. Molecular analysis revealed a novel species of Coxiella. This is believed to be the first report of a Coxiella sp. causing disease in a lorikeet. PMID- 18424843 TI - Encephalitozoon cuniculi-associated placentitis and perinatal death in an alpaca (Lama pacos). AB - Placentitis, premature birth, and perinatal death were associated with Encephalitozoon cuniculi infection in an alpaca. Histologically, chorionic trophoblasts contained many Gram-positive, period acid-Schiff positive, variably acid-fast spores. Multifocal necrosis and infiltration by lymphocytes, eosinophils, and neutrophils were scattered throughout the chorionic membrane. Spores in trophoblasts were approximately 1 microm x 2 microm, thick-walled, and contained polar filaments and polar vacuoles consistent with microsporidia. The presence of E. cuniculi DNA was confirmed by sequencing the polymerase chain reaction amplicon from frozen placental tissue. A few glial nodules were scattered throughout the cerebrum, and mild lymphocytic inflammation was present in the heart, liver, and lung. No organisms were detected in tissues other than the placenta. This is the first reported case of E. cuniculi infection in an alpaca. PMID- 18424844 TI - Characterization of a biphasic neoplasm in a Madagascar tree boa (Sanzinia madagascariensis). AB - We describe a disseminated biphasic neoplasm in a young Madagascar tree boa utilizing transmission electron and light microscopy. Discrete neoplastic cells identified within pulmonary capillaries and hepatic sinusoids represented the leukemic phase. Spindloid cells represented the sarcomatous phase, which comprised hepatic and fat body nodules. A zone of transition of the neoplastic cells, from discrete to spindloid, was noted along the periphery of the hepatic and fat body nodules. Ultrastructural examination elucidated similar nuclear features in the discrete and spindloid neoplastic cells and revealed collagen fibers within the spindloid neoplastic cells. These ultrastructural findings indicate that the discrete and spindloid cells represent a single neoplastic process with a subpopulation of cells exhibiting mesenchymal differentiation. PMID- 18424845 TI - Ocular albinism and hypopigmentation defects in Slc24a5-/- mice. AB - As part of a high-throughput mutagenesis and phenotyping process designed to discover novel drug targets, we generated and characterized mice with a targeted mutation in Slc24a5, a gene encoding a putative cation exchanger. Upon macroscopic examination, Slc24a5-/- mice were viable, fertile, and indistinguishable by coat color from their heterozygous and wild-type litter mates. Ophthalmoscopic examination revealed diffuse retinal hypopigmentation, and a histologic examination of the eye confirmed the presence of moderate-to-marked hypopigmentation of the retinal pigmented epithelium (RPE), ciliary body, and iris pigment epithelium (IPE). Hypopigmentation was most severe in the anterior layer cells of the IPE, where melanosomes were smaller, paler, and more indistinct than those of the anterior stroma and posterior IPE. The pigment granules of the posterior IPE appeared to be nearly as dark as those in stromal melanocytes; however, both cell layers were thinner and paler than corresponding layers in wild-type mice. Ultrastructural analysis of the RPE, IPE, and ciliary body pigmented cells confirmed that mutation of Slc24a5 results in marked hypopigmentation of melanosomes in optic cup-derived pigmented neuroepithelium in the eyes. Milder reductions in melanosome size and pigmentation were noted in neural crest-derived melanocytes. The severe hypopigmentation of neuroepithelium derived cells in the eyes resulted in a novel form of ocular albinism in Slc24a5 /- mice. Our findings suggest that SLC24A5 may be a candidate gene for some forms of ocular albinism and for the BEY1/EYCL2 locus previously associated with central brown eye color in humans. PMID- 18424846 TI - Postweaning multisystemic wasting syndrome in Danish pig herds: productivity, clinical signs and pathology. AB - A case-control study of 74 herds with postweaning multisystemic wasting syndrome (pmws) and 74 matched control herds was carried out. In the case herds the mortality rates of weaner and finisher pigs were 11.2 and 5.2 per cent respectively, compared with 3.1 and 3.2 per cent in the control herds. In most case herds, pmws developed within the first four weeks after weaning. Wasting, diarrhoea and respiratory signs were observed in 10 per cent of the weaner pigs (7 to 30 kg) in the case herds compared with 7 per cent in the control herds. The average daily gains of the weaner pigs and finisher pigs were 36 g and 52 g less in the case herds than in the control herds. By examining three weaner pigs from each herd the pmws diagnosis was confirmed by histopathology and immunohistochemistry in 78 per cent of the case herds, but at least one pmws positive weaner pig was found in 19 of the control herds. The prevalence of pmws positive pigs among illthriven weaner pigs was 45 per cent (101/222) in the case herds, and 12 per cent (27/222) in the control herds. Specific gross pathological findings were associated with a positive pmws diagnosis; pigs with heavy, rubber like lungs, atonic intestines, and enlarged bronchial and inguinal lymph nodes, had a 0.7 probability of a positive pmws diagnosis by laboratory examinations. However, for illthriven pigs, this probability of having pmws was equal in the case herds and the control herds. PMID- 18424847 TI - E-cadherin expression in canine cutaneous histiocytomas. AB - Canine cutaneous histiocytoma is a common skin tumour of Langerhans cell origin. Langerhans cells are members of the dendritic cell family of antigen-presenting cells and are located in the epidermis. They are unique among the dendritic cell lineage in that they express high levels of the adhesion molecule E-cadherin. The expression of E-cadherin by the neoplastic Langerhans cells in 37 dogs with cutaneous histiocytoma was studied by flow cytometry and immunohistochemistry. In all the cases, these cells expressed E-cadherin, whereas the infiltrating lymphocytes did not. PMID- 18424848 TI - Use of cardiac troponin kits for the qualitative determination of myocardial cell damage due to traumatic reticuloperitonitis in cattle. AB - This study was designed to investigate whether kits to measure circulating cardiac troponin-I (cTn-I) and cardiac troponin-T (cTn-T) can be used to determine myocardial cell damage in cattle with traumatic reticuloperitonitis (trp). Twenty cattle with trp were compared with 10 clinically healthy cattle. cTn-I and cTn-T were determined qualitatively and cTn-I was determined quantitatively; biochemical analyses were also performed on both groups. The mean serum concentrations of total protein, globulin, glucose and calcium, and the mean activities of creatine kinase mb, aspartate aminotransferase, lactate dehydrogenase and gamma-glutamyl transferase were higher in the cattle with trp than in the control group. The cTn-I and cTn-T kits both gave positive results in three of the cattle with trp and the quantitative measurement of cTn-I was positive in 11 of the trp cases. Both tests were negative in the healthy cattle. PMID- 18424849 TI - Atypical scrapie in a sheep in Scotland. PMID- 18424850 TI - Diagnostic sensitivity of nasopharyngeal and nasal swabs for the molecular detection of EHV-1. PMID- 18424851 TI - Progressive encephalomyelopathy and cerebellar degeneration in a captive-bred snow leopard (Uncia uncia). PMID- 18424852 TI - Scrapie transmission via milk. PMID- 18424853 TI - When will bluetongue transmission start in 2008? PMID- 18424855 TI - Seeking consensus on a new Veterinary Surgeons Act. PMID- 18424854 TI - Appeals against disciplinary decisions. PMID- 18424857 TI - Rumen fluke (paramphistomosis) in British cattle. PMID- 18424859 TI - Xanthophylls are preferentially taken up compared with beta-carotene by retinal cells via a SRBI-dependent mechanism. AB - The purpose of this study was to investigate the mechanisms by which carotenoids [xanthophylls vs. beta-carotene(beta-C)] are taken up by retinal pigment epithelial (RPE) cells. The human RPE cell line, ARPE-19, was used. When ARPE-19 cells were fully differentiated (7-9 weeks), the xanthophylls lutein (LUT) and zeaxanthin (ZEA) were taken up by cells to an extent 2-fold higher than beta-C (P < 0.05). At 9 weeks, cellular uptakes were 1.6, 2.5, and 3.2%, respectively, for beta-C, LUT, and ZEA. Similar extents were observed when carotenoids were delivered in either Tween 40 or "chylomicrons" produced by Caco-2 cells. Differentiated ARPE-19 cells did not exhibit any detectable beta-C 15,15' oxygenase activity or convert exogenous beta-C into vitamin A. When using specific antibodies against the lipid transporters cluster determinant 36 (CD36) and scavenger receptor class B type I (SR-BI), cellular uptake of beta-C and ZEA were significantly decreased (40-60%) with anti-SR-BI but not with anti-CD36. Small interfering RNA transfection for SR-BI led to marked knockdown of SR-BI protein expression (approximately 90%), which resulted in decreased beta-C and ZEA uptakes by 51% and 87%, respectively. Thus, the present data show that RPE cells preferentially take up xanthophylls versus the carotene by a process that appears to be entirely SR-BI-dependent for ZEA and partly so for beta-C. This mechanism may explain, in part, the preferential accumulation of xanthophylls in the macula of the retina. PMID- 18424860 TI - The pathophysiology of organophosphorus pesticide self-poisoning is not so simple. PMID- 18424861 TI - Organophosphorus pesticide poisoning: cases and developments. AB - Self-poisoning with organophosphate pesticides is a major health problem world wide. Through the inhibition of acetylcholinesterase, organophosphorus poisoning is characterised by the clinical picture of acute cholinergic crisis. Other manifestations are the intermediate neurotoxic syndrome and delayed polyneuropathy. In the Western world, the occurrence of organophosphorus poisoning is less prevalent due to the declining availability of organophosphate pesticides, which could render the recognition of this particular type of intoxication and its specific treatment more difficult. In this article we discuss some recent developments and treatment dilemmas, illustrated by cases from our clinic, followed by a review of the current recommendations in the treatment of organophosphate poisoning. PMID- 18424863 TI - Visceral involvement in an immunocompetent male: a rare presentation of cat scratch disease. AB - We report a case of an adult, immunocompetent male with lymphadenopathy of both groins, para-aortal lymph nodes and multiple lesions in the spleen. A neoplasm was excluded by histology of the largest lymph node from the left groin. The diagnosis of cat-scratch disease (CSD ) became apparent when serological testing for Bartonella henselae showed to be positive. A review of literature shows that disseminated (visceral) infection is a rare presentation of CSD. PMID- 18424862 TI - Preoperative levosimendan in heart failure patients undergoing noncardiac surgery. AB - BACKGROUND: Heart failure (HF) is a major cause of perioperative morbidity and mortality in noncardiac surgery. Preoperative optimisation of these patients is, thus, of utmost importance. Levosimendan seems promising for patients undergoing cardiac surgery; however, its safety and efficacy in HF patients undergoing noncardiac surgery have not been evaluated. OBJECTIVE: To evaluate the effects of prophylactic preoperative levosimendan administration on left ventricular function in HF patients undergoing noncardiac surgery. METHODS: HF patients with ejection fraction <30%undergoing elective noncardiac surgery in 2005 were included in this prospective study. Patients were admitted to our surgical intensive care unit one day preoperatively. Under continuous haemodynamic monitoring, the treatment protocol consisted of an initial loading dose (24 microg/kg) for ten minutes followed by a continuous 24-hour infusion (0.1 microg/kg/min) at the end of which patients underwent surgery. Echocardiography was performed before infusion (day 0) and on the 7th postinfusion day (day 7). Measurements included left ventricular ejection fraction (LVEF), velocity time integral(VTI), pre-ejection period (PEP), ejection time (ET),maximum (Pmax) and minimum P(min) transvalvular aortic pressure gradient, and maximum (Vmax) and minimum V(min) aortic velocity. RESULTS: Twelve consecutive patients were enrolled. Levosimendan resulted in a significant increase in LVEF,VTI, P(max), P(min), V(max), and V(min) (p<0.01) and, moreover, a significant reduction in PEP, ET, and PEP/ET (p=0.04) on day 7 compared with day 0 values. No adverse reactions,complications or mortality occurred during 30-day follow-up. CONCLUSION: Prophylactic preoperative levosimendan treatment may be safe and efficient for perioperative optimisation of heart failure patients undergoing noncardiac surgery. PMID- 18424864 TI - Paraganglioma of the urinary bladder. AB - Since paragangliomas of the urinary bladder are rare and not easily recognised, histological examination is often the only leading key to diagnosis. We report on a patient with a paraganglioma of the urinary bladder. Although the patient presented with classical signs and symptoms, these were only appreciated after histological examination of a transurethral resection specimen had elucidated the correct diagnosis. PMID- 18424865 TI - Acute renal failure in Plasmodium malariae infection. AB - We report an unusual case of transfusion-transmitted malaria which remained undiagnosed for several months in an Italian woman splenectomised and polytransfused for thalassaemia major. The infecting species was Plasmodium malariae, and the patient developed acute renal failure, severe thrombocytopenia, and hepatic failure. Treatment with chlorochine was followed by a slow, but complete recovery of renal function. PMID- 18424866 TI - Clinical practice guideline for cardiovascular risk management in the Netherlands. PMID- 18424867 TI - Unusual cause of chronic ascites. PMID- 18424868 TI - Erythematous pigmentation of the arm for more than ten years. PMID- 18424869 TI - Comments on the review article: Ascites in cirrhosis: a review of management and complications. PMID- 18424872 TI - Are psychological treatments for depression in primary care cost-effective? AB - BACKGROUND: Depression is a highly prevalent condition that is associated with high levels of work absenteeism and high health care costs. Most patients are treated in primary care. A large group of patients prefers psychological treatments to antidepressants. AIMS OF THE STUDY: To systematically review the evidence for the cost-effectiveness of psychological treatments, psychotherapy and counselling, in comparison with usual care or antidepressant treatment in adult primary care patients with depression. METHODS: A computer-assisted search of MEDLINE, EMBASE, CINAHL, PsycINFO, and the Cochrane Library was carried out. Two independent reviewers selected studies for the review, extracted data and assessed the methodological quality of the included studies. RESULTS: Seven studies were included in the review. Forms of psychotherapy that were evaluated were cognitive behavioural therapy, interpersonal psychotherapy and couple therapy. Usual care generally consisted of care as usually provided by the general practitioner. No conclusion can be drawn on the cost-effectiveness of the above mentioned forms of psychotherapy in comparison with usual care or antidepressant treatment. The cost-effectiveness of counselling in comparison with usual care and antidepressant therapy is yet to be established. Meta analyses showed that psychotherapy was significantly more expensive than usual care, but not significantly more expensive than antidepressant treatment. Counselling was associated with no statistically significant differences in costs and effects in comparison with usual care in the pooled analysis. DISCUSSION: Based on this review, no firm conclusions on the cost-effectiveness of psychotherapy and counselling in primary care can be drawn. Most studies had methodological shortcomings, which limit the generalisibility of the results. IMPLICATIONS FOR HEALTH POLICIES: Given the reluctance of patients to use antidepressants and the large economic impact of depression, policy makers have a need for well designed and sufficiently powered economic evaluations of psychological treatments. The available evidence seems to indicate that psychotherapy has more substantial clinical effects than counselling. Therefore, the emphasis should be on economic evaluations of forms of psychotherapy that have proved to be clinically effective. IMPLICATIONS FOR FURTHER RESEARCH: There are indications that the cost-effectiveness of depression treatment on the whole may be improved by incorporating psychological treatments into enhanced care models, tailored to the needs of individual patients and/or by providing them by trained nurses instead of psychologists or psychotherapists. Further research should investigate these patient tailored, stepped care treatment modalities for depression treatment. PMID- 18424873 TI - State mental health funding and mental health system performance. AB - BACKGROUND: The evaluation of state mental health system performance is a priority to funders, service providers and clients. AIMS: This paper tests whether state levels of mental health care expenditures are associated with state indicators of performance and access. METHODS: Data from multiple sources measured 2003 state mental health spending, on a per client and a per capita basis, and a set of 21 performance indicators. Performance measures addressed access to care, client reported quality and outcomes, employment, incarceration, homelessness, and other areas. Analyses were conducted at the state or person level depending on the data, and included bivariate correlations, multiple linear regression, and hierarchical categorical modeling. RESULTS: For 17 of 21 measures, no statistical relationship was found between performance and mental health spending, after adjusting for state income and illness severity. Spending was related to basic access measures and to lower incarceration rates, but not to the remaining measures of quality or outcome. DISCUSSION: The results suggest that expenditures through state mental health authorities in the United States do not translate to improved outcomes or quality. This may be due to ineffective administrative practices and policies which are not conducive to promoting best practices. Limitations of the data include the cross-sectional design and some weaknesses in measurement. Implications for improvement of state systems are discussed, including the need for policies that emphasize shared state-local models to implement treatment programs of known effectiveness. PMID- 18424874 TI - Costs of day hospital and community residential chemical dependency treatment. AB - BACKGROUND: Patient placement criteria developed by the American Society of Addiction Medicine (ASAM) have identified a need for low-intensity residential treatment as an alternative to day hospital for patients with higher levels of severity. A recent clinical trial found similar outcomes at social model residential treatment and clinically-oriented day hospital programs, but did not report on costs. AIMS: This paper addresses whether the similar outcomes in the recent trial were delivered with comparable costs, overall and within gender and ethnicity stratum. METHOD: This paper reports on clients not at environmental risk who participated in a randomized trial conducted in three metropolitan areas served by a large pre-paid health plan. Cost data were collected using the Drug Abuse Treatment Cost Analysis Program (DATCAP). Costs per episode were calculated by multiplying DATCAP-derived program-specific costs by each client's length of stay. Differences in length of stay, and in per-episode costs, were compared between residential and day hospital subjects. RESULTS: Lengths of stay at residential treatment were significantly longer than at day hospital, in the sample overall and in disaggregated analyses. This difference was especially marked among non-Whites. The average cost per week was USD 575 per week at day hospital, versus USD 370 per week at the residential programs. However, because of the longer stays in residential, per-episode costs were significantly higher in the sample overall and among non-Whites (and marginally higher for men). DISCUSSION: These cost results must be considered in light of the null findings comparing outcomes between subjects randomized to residential versus day hospital programs. The longer stays in the sample overall and for non-White clients at residential programs came at higher costs but did not lead to better rates of abstinence. The short stays in day hospital among non-Whites call into question the attractiveness of day hospital for minority clients. CONCLUSION: Outcomes and costs at residential versus day hospital programs were similar for women and for Whites. For non-Whites, and marginally for men, a preference for residential care would appear to come at a higher cost. IMPLICATIONS FOR HEALTH CARE PROVISION AND USE: Lengths of stay in residential treatment were significantly longer than in day hospital, but costs per week were lower. Women and Whites appear to be equally well-served in residential and day hospital programs, with no significant cost differential. Provision of residential treatment for non-Whites may be more costly than day hospital, because their residential stays are likely to be 3 times longer than they would be if treated in day hospital. For men, residential care will be marginally more costly. IMPLICATIONS FOR HEALTH POLICY FORMULATION: Residential treatment appears to represent a cost-effective alternative to day hospital for female and White clients with severe alcohol and drug problems who are not at environmental risk. IMPLICATIONS FOR FURTHER RESEARCH: The much shorter stays in day hospital than at residential among non-Whites highlight the need for research to better understand how to best meet the needs and preferences of non-White clients when considering both costs and outcomes. PMID- 18424875 TI - Differences in the cost of antidepressants across state Medicaid programs. AB - BACKGROUND: Depression is the most prevalent major mental health disorder, affecting between eight and ten percent of the population in the United States. The U.S. Medicaid programs spent in total over USD 2.3 billion on antidepressant drugs in 2003, across three categories of antidepressants including selective serotonin reuptake inhibitors, tricyclic antidepressants, and others. Each state has its own set of cost-containment policies with respect to antidepressants, as well as other drugs, including preferred drug lists, prior authorization policies, copay systems, drug utilization reviews, and physician and patient education. AIMS: Our objectives for this study are to describe in detail state Medicaid spending on antidepressants and to determine the magnitude and significance of the effects of Medicaid drug policies on reimbursement expense. METHODS: Data from the Centers for Medicare and Medicaid Services are used to calculate state expenditures on antidepressants and number of prescriptions for antidepressants. Policy variables are taken from a 2003 Kaiser Commission report. Additional data on state population, employment, and weather are found in Census 2000 and other government sources. Descriptive summary tables are used to explain reimbursement per capita (the per capita 'burden' of depression) and reimbursement per prescription. RESULTS: Per-capita reimbursement ranges from less than USD 5 in Nevada and Wisconsin to over USD 20 in Tennessee and Maine. We find that the burden of depression is heaviest in states in which the amount of annual sunshine is low; the percent of people living in rural areas is high; and the employment rate is low. Those states in which the depression burden is heaviest are those states in which cost-containment policies are pursued most vigorously. The state of Michigan has the lowest per-prescription reimbursement USD 50), followed closely by Wisconsin. Meanwhile, California, Texas, and Oklahoma have the highest reimbursement per prescription (over USD 75 in each of these states). Reimbursement per antidepressant prescription is highest in states in which the population is high; the percentage of generic prescriptions is low; and there does not exist a tiered-copay system. DISCUSSION: Of all the Medicaid policies considered, the tiered-copay system is the only policy with a statistically significant negative correlation with per-prescription cost. Since an important limitation of the study is only a single year of observation, we cannot establish the direction of causation between policy and drug cost. Another limitation of the study is that actual acquisition costs are lower than reimbursements due to manufacturer rebates. For other cost-containment programs besides cost sharing, it is the quality of the program, not its existence per se, that seems to matter. Moreover, states that have high percentages of generic drugs, regardless of policy, enjoy significantly lower costs per prescription. The results of the study also lend support to the importance of sunlight and urbanization in reducing the depression burden. IMPLICATIONS FOR POLICY AND RESEARCH: Policy makers in state Medicaid programs can learn from experiences in other states. Additional research is required to ensure that the results hold up across different years and for other therapeutic classes of drugs. PMID- 18424876 TI - Altered osmotic swelling behavior of proteoglycan-depleted bovine articular cartilage using high frequency ultrasound. AB - Swelling behavior is an electrochemical mechanical property of articular cartilage. It plays an important role in weight bearing and joint lubrication. In this study, the altered transient and inhomogeneous swelling behavior of the degenerated articular cartilage was observed and quantified in situ using ultrasound. Three groups of bovine patellar articular cartilage samples (n = 10 x 3) were obtained and digested by trypsin for 10, 20 and 30 min respectively to mimic different levels of degeneration. The osmotic-free shrinkage and swelling behavior induced by changing the concentration of the bathing saline solution from 0.15 M to 2 M and then back to 0.15 M were characterized using high frequency ultrasound (central frequency = 35 MHz) before and after digestion. It was found that the degenerated cartilage specimens showed a weaker shrinkage swelling behavior compared with the normal cartilage samples. However, no significant differences in the peak shrinkage or swelling strains were observed between different groups. The absolute values of the peak shrinkage strain significantly (p < 0.05) decreased by 45.4%, 42.1% and 50.6% respectively after the trypsin digestion for 10, 20 and 30 min, but such significance was not demonstrated for the peak swelling strains. Due to the potential alterations in the collagen-PG matrix during trypsin digestion, the correlation between the swelling strain and the shrinkage strain of the degenerated samples changed slightly in comparison with the normal samples. The proposed ultrasound method has been successfully used to measure the transient and inhomogeneous swelling behavior of the degenerated articular cartilage and has the potential for the characterization of osteoarthritis. PMID- 18424877 TI - Evaluation of a low-dose/slow-rotating SPECT-CT system. AB - The 4-slice CT that forms part of the GE Infinia Hawkeye-4 SPECT-CT scanner (Hawkeye) is evaluated against the diagnostic 16-slice CT that is incorporated in the GE Discovery ST PET-CT system (DST). The x-ray tube of the slow-rotating Hawkeye system (23 s/rotation) operates at approximately a third of the dose of diagnostic systems as used for conventional diagnostic imaging. Image reconstruction is optimized for low noise. High-contrast spatial resolution significantly falls behind diagnostic figures: the average of MTF(50) and MTF(10) (resolution where the MTF has fallen to 50% and 10%) is 2.8 +/- 0.1 cm(-1) for Hawkeye and 5.3 +/- 0.1 cm(-1) for the DST (standard reconstruction filters). Resolution in the direction of the couch movement (z coordinate) is governed by the fixed Hawkeye slice width of 5 mm. Reconstruction accuracy is found to be increased by reducing the default z increment from 4.4 mm to 2.2 mm. Low-contrast object detectability is superior compared with diagnostic systems operating in the Hawkeye dose range. In the diagnostic dose regime, however, small low contrast details remain visible in DST that are not detectable with Hawkeye. Although not of diagnostic quality, the low-dose Hawkeye provides appropriate data for SPECT attenuation correction and anatomical localization capability. PMID- 18424878 TI - Image covariance and lesion detectability in direct fan-beam x-ray computed tomography. AB - We consider noise in computed tomography images that are reconstructed using the classical direct fan-beam filtered backprojection algorithm, from both full- and short-scan data. A new, accurate method for computing image covariance is presented. The utility of the new covariance method is demonstrated by its application to the implementation of a channelized Hotelling observer for a lesion detection task. Results from the new covariance method and its application to the channelized Hotelling observer are compared with results from Monte Carlo simulations. In addition, the impact of a bowtie filter and x-ray tube current modulation on reconstruction noise and lesion detectability are explored for full scan reconstruction. PMID- 18424879 TI - EDR-2 film response to charged particles. AB - A useful tool for verifying segmental or dynamic treatments with multiple multi leaf collimator positions, spinning range modulator propellors or magnetically scanned beams would be a film with a linear dose response up to several hundred centiGray, as typical for delivered treatments. Kodak has released an extended range film (EDR-2) that may satisfy this desire. In this study, dose response curves were obtained for several electron, proton, carbon ion and iron ion beams of different energies to determine the utility of this film. PMID- 18424880 TI - Influence of the SURLAS applicator on radiation dose distributions during simultaneous thermoradiotherapy with helical tomotherapy. AB - Simultaneous thermoradiotherapy has been shown to maximize the effect of hyperthermia as a radiation sensitizer in cancer treatment. Here we follow our previous work on feasibility of thermoradiotherapy with the scanning ultrasound reflector linear array system (SURLAS) and TomoTherapy HiArt treatment system, and investigate the influence of the SURLAS hyperthermia applicator on delivered radiation dose with the TomoTherapy. A radiation treatment plan was calculated and the treatment was delivered to a phantom with SURLAS on top simulating the likely clinical setup. Proper positioning of the SURLAS was assisted with a magnetic position-and-orientation tracking device (POTD) and was verified with megavoltage-computed tomography. The delivered dose was measured with an ionization chamber (point measurement) and a radiographic film (2D dose distributions). The planned and delivered point dose data agreed within 0.61% +/- 0.63%. Planar dose data agreed within a dose difference of < or =3% of the maximum dose, and a distance-to-dose-agreement of < or =1 mm. The susceptibility of the delivered radiation dose on correct SURLAS positioning was studied as well. The largest dose discrepancy was measured for a position for which a maximum number of radiation beams intersected the incorrectly positioned SURLAS within one TomoTherapy gantry rotation. The point dose disagreed by 6.14% +/- 0.52%, and 2.25% of pixels of the 2D dose distribution did not pass the 3% dose difference/1 mm distance-to-dose-agreement criteria. Our study showed that correct positioning of the SURLAS applicator had an influence on the delivered radiation dose. Delivered and planned dose distributions were in an excellent agreement when SURLAS was positioned according to the treatment plan. Moving the applicator from its planned position was found to cause a modification of delivered dose distributions. A precise and reproducible positioning of the applicator was assured with a POTD. PMID- 18424881 TI - Regularization of DT-MR images using a successive Fermat median filtering method. AB - Tractography using diffusion tensor magnetic resonance imaging (DT-MRI) is a method to determine the architecture of axonal fibers in the central nervous system by computing the direction of greatest diffusion in the white matter of the brain. To reduce the noise in DT-MRI measurements, a tensor-valued median filter, which is reported to be denoising and structure preserving in the tractography, is applied. In this paper, we proposed the successive Fermat (SF) method, successively using Fermat point theory for a triangle contained in the two-dimensional plane, as a median filtering method. We discussed the error analysis and numerical study about the SF method for phantom and experimental data. By considering the computing time and the image quality aspects of the numerical study simultaneously, we showed that the SF method is much more efficient than the simple median (SM) and gradient descents (GD) methods. PMID- 18424882 TI - Measuring brain hemodynamic changes in a songbird: responses to hypercapnia measured with functional MRI and near-infrared spectroscopy. AB - Songbirds have been evolved into models of choice for the study of the cerebral underpinnings of vocal communication. Nevertheless, there is still a need for in vivo methods allowing the real-time monitoring of brain activity. Functional Magnetic Resonance Imaging (fMRI) has been applied in anesthetized intact songbirds. It relies on blood oxygen level-dependent (BOLD) contrast revealing hemodynamic changes. Non-invasive near-infrared spectroscopy (NIRS) is based on the weak absorption of near-infrared light by biological tissues. Time-resolved femtosecond white laser NIRS is a new probing method using real-time spectral measurements which give access to the local variation of absorbing chromophores such as hemoglobins. In this study, we test the efficiency of our time-resolved NIRS device in monitoring physiological hemodynamic brain responses in a songbird, the zebra finch (Taeniopygia guttata), using a hypercapnia event (7% inhaled CO(2)). The results are compared to those obtained using BOLD fMRI. The NIRS measurements clearly demonstrate that during hypercapnia the blood oxygen saturation level increases (increase in local concentration of oxyhemoglobin, decrease in deoxyhemoglobin concentration and total hemoglobin concentration). Our results provide the first correlation in songbirds of the variations in total hemoglobin and oxygen saturation level obtained from NIRS with local BOLD signal variations. PMID- 18424883 TI - Preoperative paclitaxel/carboplatin radiochemotherapy for stage III/IV resectable oral and oropharyngeal cancer: seven-year follow-up of a phase II trial. AB - Numerous treatment concepts for advanced but resectable oral and oropharyngeal squamous cell carcinoma exist. In this study, we present the 7-year results of a promising treatment with preoperative simultaneous chemoradiation using paclitaxel and carboplatin within a prospective phase II trial comprising 56 patients. After determination of the local tumor extension, chemoradiation was applied for 4 weeks and up to 40 Gy. Following a recovery period of 3-4 weeks, tumor resection was performed within the initially tattooed resection margins, together with a functional modified neck dissection. The median follow-up time was 44.9 +/- 19.6 months (range 0.76-87.9). After 7 years, 35 (63.3%) patients were alive and 20 (36.4%) had died. In 2 patients (3.6%), the cause of death was related to treatment. After 7 years, the overall survival rate declined to 63.6%, whereas the local recurrence-free probability was still 84.2%. These results confirm the excellent local control and high survival rates of preoperative radiochemotherapy with the combination of paclitaxel/carboplatin. PMID- 18424884 TI - Phase II trial of pegylated liposomal doxorubicin-cyclophosphamide combination as first-line chemotherapy in older metastatic breast cancer patients. AB - OBJECTIVE: To evaluate the efficacy and toxicity of pegylated liposomal doxorubicin (PLD; Caelyx)-cyclophosphamide combination in older metastatic breast cancer patients. METHODS: A multicenter phase II trial was conducted. Inclusion criteria were age 65-75 years, ECOG 0-1 and left ventricular ejection fraction > or =50%. First-line chemotherapy was given to metastatic breast cancer patients resistant to hormonal therapy. The treatment schedule was PLD 40 mg/m(2) and cyclophosphamide 500 mg/m(2) on day 1 every 4 weeks. Efficacy was the primary endpoint, while response duration and tolerance were the secondary endpoints. RESULTS: Thirty-five patients (median age 71.3 years) were enrolled. No treatment related death, no congestive heart failure or decrease in left ventricular ejection fraction and no febrile neutropenia were reported. TOXICITY: grade 3 dyspnea was found in 1 patient, neutropenia in 11 patients (7 grade 3, 4 grade 4), grade 3 mucositis in 4 patients, grade 3 hand-foot syndrome in 1 patient and a generalized rash in 1 patient. An objective response (complete and partial response) was achieved in 10 (28.6%) patients and disease control in 24 (69%) with a progression-free survival of 8.8 months and a median overall survival of 20.3 months. CONCLUSION: The PLD-cyclophosphamide combination is moderately active and safe in elderly metastatic breast cancer patients, but cannot be recommended routinely due to myelotoxicity and mucositis hazards. PMID- 18424885 TI - The baseline ratio of neutrophils to lymphocytes is associated with patient prognosis in advanced gastric cancer. AB - OBJECTIVE: In cancer patients, the balance between neutrophil (N) and lymphocyte (L) cell counts fluctuates with advancing disease. The objective of our study was to determine the prognostic implications of the N/L ratio in the peripheral blood of gastric cancer patients. METHODS: Study participants were identified from a prospective cohort of patients with advanced gastric cancer in Japan (n = 1,220). RESULTS: The median baseline N/L was 2.58 (range, 0.63-12.7). Univariate analysis revealed that patients with an N/L > pr =2.5 (n = 644) had a significantly poorer prognosis than those with an N/L <2.5 (n = 576; log rank test, p = 0.019 x 10( 12)). The median survival times for these two groups were 239 (95% confidence interval, CI, 217-251 days) and 363 days (95% CI, 334-406 days), respectively, while the 1-year survival rates were 30 (95% CI, 26-34%) and 50% (95% CI, 45 54%), respectively. A multivariate Cox model established a significant relationship between the N/L ratio and survival (adjusted hazard ratio = 1.52; 95% CI, 1.32-1.75; p = 0.077 x 10(-8)). CONCLUSIONS: These results suggest that the N/L ratio is an independent prognostic factor in advanced gastric cancer. Measurement of this ratio may serve as a clinically accessible and useful biomarker for patient survival. PMID- 18424886 TI - Oral capecitabine in gemcitabine-pretreated patients with advanced pancreatic cancer. AB - OBJECTIVE: To date, no standard regimen for salvage chemotherapy after gemcitabine (Gem) failure has been defined for patients with advanced pancreatic cancer (PC). Oral capecitabine (Cap) has shown promising activity in first-line chemotherapy trials in PC patients. METHODS: Within a prospective single-center study, Cap was offered to patients who had already received at least 1 previous treatment regimen containing full-dose Gem (as a single agent, as part of a combination chemotherapy regimen or sequentially within a chemoradiotherapy protocol). Cap was administered orally at a dose of 1,250 mg/m(2) twice daily for 14 days followed by 7 days of rest. Study endpoints were objective tumor response rate by imaging criteria (according to RECIST), carbohydrate antigen 19-9 (CA19 9) tumor marker response, time to progression, overall survival and toxicity. RESULTS: A median of 3 treatment cycles (range 1-36) was given to 39 patients. After a median follow-up of 6.6 months, 27 patients were evaluable for response: no complete or partial responses were observed, but 15 patients (39%) had stable disease. A CA19-9 reduction of >20% after 2 cycles of Cap was documented in 6 patients (15%). Median time to progression was 2.3 months (range 0.5-45.1) and median overall survival (since start of Cap treatment) was 7.6 months (range 0.7 45.1). Predominant grade 2 and 3 toxicities (per patient analysis) were hand-foot syndrome 28% (13% grade 3); anemia 23%; leg edema 15%; diarrhea 13%; nausea/vomiting 10%, and leukocytopenia 10%. CONCLUSION: Single-agent Cap is a safe treatment option for Gem-pretreated patients with advanced PC. Further evaluation of Cap in controlled clinical trials of Gem-pretreated patients with advanced PC is recommended. PMID- 18424887 TI - Capecitabine and vinorelbine as first-line treatment in elderly patients (> or =65 years) with metastatic breast cancer. A phase II trial (SAKK 25/99). AB - BACKGROUND: We evaluated previously established regimens of capecitabine plus vinorelbine in older patients with advanced breast cancer stratified for presence versus absence of bone metastases. PATIENTS AND METHODS: Patients > or =65 years who had received no prior chemotherapy for advanced breast cancer received up to six 21-day cycles of vinorelbine 20 mg/m(2) i.v. on days 1 + 8 with oral capecitabine on days 1-14 (1,000 vs. 1,250 mg/m(2) daily in patients with vs. without bone involvement). RESULTS: Median age was 72 years in patients with bone metastases (n = 47) and 75 years in patients without bone metastases (n = 23). Response rates were 43% (95% confidence interval, CI, 28.3-58.8) and 57% (95% CI = 34.5-76.8), respectively. Median time to progression was 4.3 (95% CI = 3.5-6.0 months) and 7.0 months (CI = 4.1-8.3), respectively. Neutropenia was the most common toxicity, with grade 3/4 occurring in 43 and 39%, respectively. Pulmonary embolism was seen in 5 and grade 3 thrombosis in 3 patients. Other toxicities were mild to moderate. CONCLUSIONS: These regimens of capecitabine and vinorelbine are active and well tolerated in patients with advanced breast cancer > or =65 years. Response rates were comparable to published results. The lower capecitabine doses appeared appropriate given the advanced age, bone involvement and prior radiotherapy. PMID- 18424888 TI - Loss of expression of EphB1 protein in gastric carcinoma associated with invasion and metastasis. AB - OBJECTIVE: EphB1 is a member of the Eph family of receptor tyrosine kinases that is involved in embryonic nervous and vascular system development. Over- or underexpression of certain Eph receptors has been found in some cancer samples compared to normal tissue. Expression of Eph receptors is related to malignant transformation, metastasis, differentiation, and prognosis of cancers. Recently, the EphB subfamily has been shown to be involved in the tumorigenesis of colorectal cancer. In the present study, expression of the EphB1 transcript and protein in gastric carcinoma samples was determined to investigate the roles of EphB1 in development, progress and prognosis of gastric carcinoma. METHODS: Quantitative real-time reverse transcriptase-polymerase chain reaction and immunohistochemical staining were used. RESULTS: The EphB1 transcript was overexpressed in 68.9% (42/61) and underexpressed in 14.8% (9/61) of cases. However, the expression of protein was greatly different from the transcript expression, with overexpression and underexpression being 17.2% (10/58) and 44.8% (26/58), respectively. In addition, we showed that underexpression of EphB1 protein is significantly associated with invasion, stage and metastasis in gastric carcinomas. CONCLUSION: EphB1 may have a tumor-suppressive role in gastric cancer. PMID- 18424889 TI - Expression of interferon-gamma-inducible protein 10 related to angiogenesis in uterine endometrial cancers. AB - OBJECTIVE: Angiogenesis is essential for the development, growth and advancement of solid tumors. Interferon-gamma-inducible protein 10 (IP-10) regulates lymphocyte chemotaxis, mediates vascular pericyte proliferation and acts as an angiostatic agent, thus inhibiting tumor growth. This prompted us to study the clinical implications of IP-10 expression related to angiogenesis in uterine endometrial cancers. METHOD: Sixty patients underwent curative resection for uterine endometrial cancers. In the tissue of these cancers, the levels of IP-10, vascular endothelial growth factor, interleukin-8 and basic fibroblast growth factor (bFGF) were determined by enzyme immunoassay, and the localization of IP 10 and counts of microvessels were determined by immunohistochemistry. RESULT: IP 10 is diffusely localized in the cancer cells, but not in the stromal cells. There was a significant, reverse correlation between microvessel counts and IP-10 levels in uterine endometrial cancers. The IP-10 levels significantly decreased with more advanced disease and significantly reverse-correlated with bFGF levels in uterine endometrial cancers. CONCLUSIONS: IP-10 might affect the suppression of angiogenesis associated with bFGF in advanced cancer. Furthermore, IP-10 activation might be effective in the suppression of regrowth or recurrence after intensive treatment for advanced endometrial cancers. PMID- 18424890 TI - Altered gene expression in rat colonic adenocarcinomas induced in an azoxymethane plus 2-amino-1-methyl-6-phenylimidazo[4,5-b]- pyridine initiation-promotion model. AB - 2-Amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP), the most abundant food derived mutagenic/carcinogenic heterocyclic amine (HCA), has attracted particular attention as a probable human colon carcinogen. Some studies have shown that PhIP administered in the post-initiation phase is able to enhance rat colon carcinogenesis remarkably. To determine whether this genotoxicant leaves a DNA footprint in colon carcinogenesis, 6-week-old male F344 rats were first subcutaneously injected with azoxymethane (AOM) and then continuously treated with various doses (0-200 ppm) of PhIP added to their diet. Animals were killed at week 36 for histopathological examination, and colonic adenocarcinomas derived from animals receiving 0, 50 and 200 ppm PhIP were subjected to a novel three dimensional (3D)-microarray and real-time reverse transcription-polymerase chain reaction (RT-PCR) analysis. A total of five candidate genes were identified in adenocarcinomas following 200 ppm of PhIP and AOM initiation, with a dose dependent increment. Among them, Stat1 (signal transducer and activator of transcription 1) and VEGFc (vascular endothelial growth factor c) demonstrated statistically significant upregulation by real-time RT-PCR. In addition, HSP90 (heat shock protein 90) and VEGFa showed a non-significant tendency to increase. In summary, overexpression of Stat1, VEGF and other genes could be involved in PhIP-enhanced colon tumorigenesis in the post-initiation phase. PMID- 18424891 TI - Circulating adiponectin levels and expression of adiponectin receptors in relation to lung cancer: two case-control studies. AB - BACKGROUND: Decreased circulating levels of adiponectin, an adipocyte-secreted hormone and endogenous insulin sensitizer, have been associated with several obesity-related malignancies. Thiazolidinedione administration, which increases adiponectin levels, decreases risk for lung cancer. Whether circulating adiponectin levels are associated with lung cancer and/or whether adiponectin receptors are expressed in lung cancer remains unknown. METHODS: We conducted a case-control study of 85 patients with incidental, histologically confirmed lung cancer and 170 healthy controls matched by gender and age. In a separate study, archival lung specimens from 134 cancerous and 8 noncancerous tissues were examined for relative expression of adiponectin receptors AdipoR1 and AdipoR2 using immunohistochemistry. RESULTS: Tobacco smoking, heavy alcohol intake and education were all associated with lung cancer risk, whereas serum adiponectin levels were not significantly different between cases and controls (multiple logistic regression, odds ratio per SD of adiponectin among controls: 1.13, 95% confidence interval: 0.64-2.02). Adiponectin levels were significantly lower (odds ratio: 0.25, 95% confidence interval: 0.10-0.78) among patients with advanced compared to those with limited disease stage. Expression of adiponectin receptors was apparent only in the cancerous lung tissue (64.2% AdipoR1 and 61.9% AdipoR2 in cancerous vs. 0% among noncancerous tissue). Specifically, AdipoR1 was expressed in all disease types, but no difference was noted with disease stage, whereas AdipoR2 was mainly expressed in the non-small cell carcinomas and more prominently in the advanced disease stage (80%). CONCLUSIONS: Circulating adiponectin levels are not different in cases of this malignancy - which seems to be unrelated to obesity and insulin resistance - compared to their healthy controls, though hormonal levels were significantly lower in advanced versus limited lung cancer. Both adiponectin receptors were expressed in cancerous lung tissue, but not in normal control tissue and there was a differential expression by disease stage. These findings should be further explored, especially in the context of the recently reported protective effect of thiazolidinediones in diabetic patients with lung cancer. PMID- 18424892 TI - Pulmonary fibrosis after chemotherapy with oxaliplatin and 5-fluorouracil for colorectal cancer. AB - BACKGROUND: Chemotherapy with oxaliplatin and 5-fluorouracil (5-FU)/folinic acid is the standard first-line therapy of metastatic colorectal carcinoma and has shown activity in several other malignancies. This regimen is mostly well tolerated. Known side effects include myelosuppression, nausea/vomiting and neuropathies; acute pulmonary toxicity has only been described in very few reports. CASE REPORT: A 66-year-old male with metastatic rectal adenocarcinoma treated with 12 cycles of oxaliplatin and 5-FU/folinic acid developed bilateral pulmonary infiltrates and respiratory failure. Broad-spectrum antibiotic therapy did not improve his condition and extended microbiological diagnostics did not show an infectious etiology. Therapy with corticosteroids led to a short improvement, however the patient died 1 week after the initiation of corticosteroid treatment due to respiratory insufficiency. The clinical and histopathological data as well as the lack of an infectious cause indicate that pulmonary fibrosis was induced by oxaliplatin and 5-FU/folinic acid. CONCLUSION: This case demonstrates that treatment with oxaliplatin and 5-FU/folinic acid can cause acute pulmonary fibrosis. Even though pulmonary toxicity is rare in patients treated with this chemotherapy regimen compared to infectious pulmonary complications, it should be considered early in the clinical course of otherwise unexplained pulmonary infiltrates hopefully leading to a better outcome. PMID- 18424893 TI - Rapid manifestation of CNS metastatic disease in a cervical carcinoma patient: a case report. AB - OBJECTIVES: Brain metastases originating from primary cervical carcinoma are very rare. Despite surgery and adjuvant therapy, patient prognosis is extremely unfavorable. Prior studies have reported that the median time from the primary disease's diagnosis to development of CNS metastases is approximately 18 months. CASE REPORT: A 60-year-old woman was initially diagnosed and treated for FIGO clinical stage IB2 cervical carcinoma in March 2007. She underwent a radical hysterectomy, bilateral salpingo-oophorectomy and peri-aortic lymphadenectomy. Two weeks later, the patient presented with dysmetria and homonymous hemianopsia, which was related to a large right occipital CNS metastasis. The patient had surgery to resect the brain tumor and then received Gamma Knife stereotactic radiosurgery and chemotherapy. The neurological deficits were resolved and the patient's disease status has since remained stable, with five months of follow up. CONCLUSION: The incidence of brain metastases originating from primary cervical carcinoma is very rare. Furthermore, the occurrence of CNS metastatic disease manifesting itself within a couple weeks following diagnosis of the primary disease is even more unusual. Since the presenting symptoms of CNS metastatic disease are not well characterized and patient prognosis is very poor, oncology physicians should anticipate the presence of this condition in order to provide prompt and comprehensive treatment. PMID- 18424894 TI - Pyridoxine as prophylactic therapy for palmar-plantar erythrodysesthesia associated with administration of pegylated liposomal doxorubicin (caelyx): a single-center experience. PMID- 18424895 TI - Organic elemental composition in fingernail plates varies between sexes and changes with increasing age in healthy humans. AB - BACKGROUND: Keratin, an alpha-helical fibrous protein, is the primary component of human nail plates. No data on age-related changes in healthy subjects are present. OBJECTIVE: This study investigated whether keratin amount and composition, as indicated by organic elemental composition of fingernails, varies with aging and between sexes. METHODS: Nail clippings from 225 healthy individuals (93 males, 132 females), aged 20-90 years, were analyzed for carbon (C), nitrogen (N), and sulfur (S) contents with an automatic elemental analyzer. C/N and N/S ratios were calculated. The C/N ratio is indicative of keratin composition, i.e. the ratio of alpha-amino acids and protein. The relationship of elemental composition with dietary intake was analyzed by standardized food record. RESULTS: Females have in their nails more sulfur (p < 0.001) and less nitrogen (p = 0.001), and thereby, a lower N/S ratio than males, whereas their carbon content is not different. With aging, the carbon content increases (p < 0.01, both sexes) and the nitrogen content decreases (p = 0.05, females), both leading to an increased C/N ratio (p < 0.001). By contrast, the sulfur content and the N/S ratio do not change with aging (p > 0.05). The carbon content correlates positively with macronutrient intake in females. Nitrogen and sulfur contents are not related with dietary intake. CONCLUSIONS: Results suggest that the N/S ratio is indicative of sex differences and the C/N ratio of aging in healthy humans. The increasing carbon content with ongoing age could be explained by loss of inorganic material from the nails, followed by a subsequent increase of organic material. The increasing C/N ratio gives evidence that keratin composition changes towards a higher amount of alpha-amino acids with aging. PMID- 18424896 TI - Insufficient tetanus vaccination status in patients with chronic leg ulcers. Results of a prospective investigation in 100 patients. AB - BACKGROUND: Tetanus disease is caused by Clostridium tetani and is one of the most common infectious diseases worldwide. Despite international recommendations for patients with a chronic leg ulcer, there has been a distinctive lack of protection provided by vaccination for these patients in the past decades. METHODS: Within the context of our prospective clinical investigation we consecutively determined the concentrations of immunoglobulin G antibodies against C. tetani in 100 patients with a chronic leg ulcer between January 2005 and November 2006. RESULTS: A total of 38 patients were male, and 62 were female. Their mean age was 71 years (25-94). In a total of 47% (n = 47; 13 male, 34 female, mean age: 76 years) of the patients, insufficient immunoglobulin G antibody concentrations were detected. Particularly the subanalysis indicated an insufficient tetanus protection provided by vaccination in 70% of the people aged >or=80 years. CONCLUSION: A chronic wound, e.g. in the form of a leg ulcer, is known as a potential entrance for C. tetani. Unlike acute wounds, however, it is hardly ever considered to be a reason for assessment of the tetanus immune status. The results of our investigation clarify that particularly elderly people suffering from a leg ulcer have to be tested for tetanus protection provided by vaccination more strictly than ever, and if necessary, vaccinations have to be renewed. PMID- 18424897 TI - Skin cancer knowledge and preventive behaviors among patients with a recent history of cutaneous squamous cell carcinoma. AB - AIMS: To evaluate skin cancer knowledge and preventive behaviors of patients recently treated for cutaneous squamous cell carcinoma (SCC) and to examine the factors associated with the adoption of preventive behaviors. METHODS: Telephone survey on 315 SCC patients treated at a large dermatological hospital in Italy, evaluating skin cancer knowledge, sun protection and skin examination practices as well as medical recommendations received after SCC removal. RESULTS: Skin cancer knowledge was fair/low for 48.9% of the participants. Doctors were the main source of skin cancer information for 24.4% of the patients. Of the patients assessed >or=12 months after SCC removal, 32.7% reported a total skin examination after removal. Of the participants, 41.6% never/rarely used sunscreens. In a multivariate analysis, the likelihood of having complete skin examinations was associated with a doctor's advice to have an examination (odds ratio, OR = 2.29; 95% confidence interval, CI = 1.2-4.4), a higher knowledge level (OR = 2.05; 95% CI = 1.1-3.8) and past skin examinations (OR = 3.62; 95% CI = 1.9-7.0). Doctor's recommendations increased the likelihood of adopting preventive behaviors. CONCLUSIONS: We found substantial knowledge gaps and limited adoption of skin cancer prevention, highlighting the need for interventions promoting knowledge and preventive behaviors, particularly among higher-risk patients. PMID- 18424898 TI - Relationship between smiling and laughter in humans (Homo sapiens): testing the power asymmetry hypothesis. AB - The power asymmetry hypothesis claims that individuals should have distinct signals of appeasement/affiliation and play when status difference is high, whereas these signals should overlap in egalitarian interactions. Naturalistic observations were conducted on humans interacting in groups that differed in terms of age composition (and presumably social status). Three affiliative behaviours were recorded by focal sampling: spontaneous smiles, deliberate smiles and laughter. Interestingly, young men showed significantly higher proportions of deliberate smiles in comparison to laughter when interacting with people of a different age class than when interacting in same-age groups. The pattern of affiliative behaviours in women remained unaffected by the age composition of groups. This partly supports the power asymmetry hypothesis and suggests that in men, deliberate smiles could play a role in the regulation of hierarchical relationships. PMID- 18424899 TI - Cardiothoracic ratio, malnutrition, inflammation, and two-year mortality in non diabetic patients on maintenance hemodialysis. AB - BACKGROUND/AIMS: Cardiothoracic ratio (CTR) is associated with left ventricular mass, left ventricular systolic dysfunction and mortality in patients with hypertension or chronic cardiovascular disease. The clinical significance of CTR in non-diabetic maintenance hemodialysis (MHD) patients remains unclear. METHODS: 468 non-diabetic MHD patients were enrolled. Geographic, hematological, biochemical and dialysis-related data were obtained. The patients were analyzed for nutritional and inflammatory markers as well as CTR. All patients were followed up for 2 years to investigate the risks for mortality. RESULTS: Chi square analysis showed that the incidence of malnutrition and inflammation was significantly higher in patients with CTR >60% than in patients with CTR 50%. PMID- 18424900 TI - Measuring the prevalence of stroke. PMID- 18424901 TI - Homocysteine, folate and cognition in a large community-based sample of elderly people--the 3C Dijon Study. AB - BACKGROUND: Hyperhomocysteinemia is associated with an increased risk of cognitive impairment in the elderly. Recent studies suggest that folate level may also influence the course of cognitive decline. OBJECTIVE: We performed the cross sectional analysis of the relationship between homocysteine and folate levels and cognitive performances in a population-based study including 3,914 subjects aged 65 years and older. METHOD: Subjects had an evaluation of their cognitive level using five neuropsychological tests. A Cognitive Summary Score was computed as the sum of each of the 5 cognitive tests score standardized. Relations of folate and homocysteine levels with cognition were first studied separately in a covariance analysis. Stratified analyses were also performed because of interaction between folate and homocysteine in relation to cognition. RESULTS: Subjects in the higher quartile of homocysteine (high homocysteine group) and subjects in the lower quartile of folate (low folate group) had consistently lower cognitive performances in all tests. When stratified on folate level, high homocysteinemia was associated with lower cognitive performances only in subjects with a low folate level. CONCLUSION: In this large population-based sample of elderly people, the association between high homocysteinemia and decreased cognition was only seen in participants with low folate levels. PMID- 18424902 TI - Creutzfeldt-Jakob disease in Austria: an autopsy-controlled study. AB - BACKGROUND: Definite diagnosis of prion diseases or transmissible spongiform encephalopathies (TSEs) requires neuropathology, usually at autopsy. Epidemiology of human TSEs has relied on definite as well as 'probable' cases in which neuropathological confirmation is lacking, usually because of low autopsy rates in most countries. METHODS: In Austria, an active surveillance program for human prion diseases was established in 1996. Since then, more than 900 referrals were analyzed. Postmortem investigation of brain tissue is mandatory in every suspect case of TSE. Thus, epidemiological data on TSEs from Austria may serve as autopsy controlled reference for countries with lower autopsy rates. RESULTS: The total number of TSE cases in Austria since 1969 is 206. The average yearly mortality for the active surveillance period from 1996 to 30 June 2006 is 1.39 per million, with the highest rates for Vienna (2.37) compared with other provinces. Eighty five percent of definite TSEs were classified as sporadic Creutzfeldt-Jakob disease (sCJD). We observed a significant linear increase in the mean age at death of 0.6 years per calendar year. Clinical diagnostic surveillance criteria had a sensitivity and specificity of 82.7 and 80.0% for probable CJD, respectively, and a positive predictive value of 80.5% for probable and 38.9% for 'possible' CJD. Alternative neuropathological diagnoses in suspect cases included Alzheimer's disease with or without Lewy body pathology, vascular encephalopathy, metabolic encephalopathies and viral or limbic encephalitis. CONCLUSION: The steady increase in mortality rates, especially in old age groups, most likely reflects improved case ascertainment due to active surveillance causing higher awareness of the medical community. In comparison with other European countries, it is reassuring to note that the overall death rate of TSEs does not differ from the Austrian autopsy-controlled data, thus confirming the value of clinical surveillance criteria. PMID- 18424903 TI - Spatial clusters of Creutzfeldt-Jakob disease mortality in Japan between 1995 and 2004. AB - BACKGROUND: There is suggested to be a geographical difference in Creutzfeldt Jakob disease (CJD) mortality in Japan. We performed a study to detect localized clusters and hot-spot areas of deaths from CJD in Japan during the 10-year period from 1995 to 2004. METHODS: The diagnosis of CJD was taken from the death certificate (coded as A81.0 in the ICD-10). A total number of 1,168 CJD deaths (500 males and 668 females) were used for analysis using empirical Bayes estimates of standardized mortality ratios and the flexible spatial scan statistic to detect clusters. To detect the most likely cluster, p values were obtained using Monte Carlo hypothesis testing (with p < 0.05 as statistical significance). RESULTS: The most likely cluster of CJD mortality was located in the northwest region from the base of Mt. Fuji, stretching over the two neighboring prefectures of Yamanashi and Shizuoka (relative risk = 2.28, p = 0.021). Some other clusters were detected but were not significant. CONCLUSIONS: The present study supports the evidence of geographical clustering of deaths from CJD at a specific location in Japan. PMID- 18424904 TI - Association of the homeobox transcription factor gene ENGRAILED 2 with autistic disorder in Chinese children. AB - BACKGROUND: Autism is a neurodevelopmental disorder with a strong genetic component. Previous studies have mapped the disease to chromosome 7q, where the homeobox transcription factor ENGRAILED 2 (EN2) gene is located. EN2 is specifically involved in patterning the region that gives rise to the cerebellum. In the present work, we carried out a case-control study to determine whether 2 intronic single-nucleotide polymorphisms (SNPs) of EN2 are a susceptibility to autism in a Han Chinese population. METHOD: We enrolled 184 cases of DSM-IV-TR diagnosed autistic disorder, 225 controls of unrelated healthy volunteers and 409 randomly selected controls from the community who lives in the adjacent geographical regions for this study. Two SNPs (rs1861972, rs1861973) at the EN2 gene that have been reported to be associated with autism underwent analysis among our studied cohorts. Both the UNPHASE and PHASE statistical programs were utilized for evaluating the association of EN2 SNPs with autism based on allelic and genotypic frequencies and haplotype compositions accompanied with the goodness-of-fit method of the chi(2) test. The gender difference was also investigated by using 2-side Fisher's exact test treated as a covariate in logistic regression analysis. RESULTS AND CONCLUSION: Both the allelic and genotypic distributions of the 2 polymorphisms were concordant with Hardy Weinberg equilibrium. Significant differences were found for cases versus community and overall controls. By using the UNPHASE and PHASE programs, the 2 marker haplotype A-C of EN2 was identified to have a protective effect for autism, indicating that the ethnic difference might confound the EN2 association with autism. Therefore, more EN2 gene association studies of Han Chinese populations are warranted to confirm this finding. PMID- 18424905 TI - Dipole source analysis of temporal slow waves in the elderly. AB - BACKGROUND: Temporal low-voltage irregular delta-waves (TLID) are often found in elderly subjects. The physiological significance of TLID has not been clarified; however, our previous studies suggest that TLID are associated with mild cerebrovascular dysfunction. OBJECTIVE: The present study aimed to reveal the origin of TLID and their neural mechanisms by dipole source modeling. METHODS: From electroencephalography records taken from 21 scalp electrodes, clear and typical TLID of 6 elderly subjects (mean age, 69 +/- 6.2 years) were selected. Among these, we selected and averaged 7-12 clear TLID on the left side in each subject, and estimated a single equivalent current dipole for the averaged TLID. RESULTS: The best equivalent current dipoles were estimated to be located in the medial part of the temporal lobe in or near the parahippocampal gyrus in the hemisphere ipsilateral to the TLID, with a high reliability in all subjects. CONCLUSIONS: Considering the source localization of TLID, TLID seem to indicate certain dysfunctions of the hippocampus or adjacent regions. This is the first study to report the cerebral origin of TLID and suggest its physiological significance. PMID- 18424906 TI - Diurnal cortisol levels and cortisol response in youths with obsessive-compulsive disorder. AB - BACKGROUND/AIMS: Recent results indicate a role of the hypothalamic-pituitary adrenal (HPA) axis in the pathophysiology of obsessive-compulsive disorder (OCD). Although childhood onset is common, the HPA axis has scarcely been studied in young OCD subjects. Therefore, the present study aimed at examining basal and response levels of salivary cortisol in a sample of young OCD subjects. METHODS: Twenty-three children and adolescents with DSM-IV OCD were compared to a reference group of school children (n = 240-336). The basal cortisol rhythm was measured through saliva samples 3 times/day. The cortisol response to a psychological stressor (exposure therapy in the OCD group and a fire alarm in the reference group) was also examined. RESULTS: Compared to the reference group, OCD subjects displayed higher early-morning cortisol values (p = 0.005) with no difference between the late-morning and evening values. The cortisol levels in the OCD group diminished in response to the psychological stressor, compared to a positive response in the reference group (p < 0.001). No relation was found between cortisol and clinical parameters. CONCLUSION: These results support the idea that HPA hyperactivity, commonly found in adult OCD patients, is also present at an earlier stage of development, with specificity for the early morning peak. PMID- 18424907 TI - Val158Met polymorphism in the catechol-O-methyltransferase (COMT) gene is not associated with tardive dyskinesia in schizophrenia. AB - OBJECTIVE: This study investigated whether the catechol-O-methyltransferase (COMT) gene V158M single-nucleotide polymorphism (SNP) influences susceptibility to tardive dyskinesia (TD). METHODS: We examined 209 Korean schizophrenic patients using the Abnormal Involuntary Movement Scale (AIMS), with genotyping performed for the COMT gene V158M SNP. RESULTS: The logistic regression analysis showed that old age [p = 0.032, OR = 1.40 (OR corresponds to 10-year), 95% CI = 1.03-1.90] was a risk factor for TD, but there was no significant association between the COMT genotype and TD. The heterozygotes (MV genotype) of the COMT gene polymorphism tended to develop TD less than homozygotes (MM and VV); however, the risk did not reach statistical significance (p = 0.050, OR = 1.81, 95% CI = 1.00-3.29). CONCLUSIONS: These results suggest that the V158M SNP of the COMT gene is not associated with TD in schizophrenia. However, there is a tendency that the heterozygous genotype of the COMT gene polymorphism has a protective effect against TD. Further investigations are warranted to evaluate a molecular heterosis of this polymorphism in development of TD in a large sample of subjects. PMID- 18424908 TI - Tobacco smoking and depression--results from the WHO/ISBRA study. AB - AIMS: To elucidate the relationship between tobacco smoking and depression, and to estimate the impact of other substance dependencies. DESIGN: Cross-sectional. PARTICIPANTS: A total of 1,849 men and women were interviewed face-to-face using a validated structured questionnaire. According to their tobacco smoking behavior, participants were grouped into never smokers, ex-smokers and current smokers. MEASUREMENTS: Data were generated through the WHO/ISBRA study, an international multicenter study with a cross-sectional design. A standardized questionnaire was administered face-to-face by trained interviewers. Logistic regression analysis was used to predict depression. RESULTS: There was a significant difference across the 3 smoking groups in the number of subjects who had major depression (DSM-IV) during their lifetime. The highest rate of depressives was found in current smokers (23.7%), the lowest rate in never smokers (6.2%), while the rate of those who had quit smoking (14.6%) was between both. In a logistic regression analysis, alcohol dependence (both current and during lifetime) as well as cocaine dependence were significant predictors of depression. However, the association between smoking and depression still remained statistically significant. CONCLUSIONS: This study adds support to the evidence that smoking is linked to depression. It also elucidates the importance of taking into account alcohol and cocaine dependence since they have a significant impact on the relationship between smoking and depression. PMID- 18424909 TI - A UDP-glucose derivative is required for vacuolar autophagic cell death. AB - Autophagic cell death in Dictyostelium can be dissociated into a starvation induced sensitization stage and a death induction stage. A UDP-glucose pyrophosphorylase (ugpB) mutant and a glycogen synthase (glcS) mutant shared the same abnormal phenotype. In vitro, upon starvation alone mutant cells showed altered contorted morphology, indicating that the mutations affected the pre death sensitization stage. Upon induction of cell death, most of these mutant cells underwent death without vacuolization, distinct from either autophagic or necrotic cell death. Autophagy itself was not grossly altered as shown by conventional and electron microscopy. Exogenous glycogen or maltose could complement both ugpB(-) and glcS(-) mutations, leading back to autophagic cell death. The glcS(-) mutation could also be complemented by 2-deoxyglucose that cannot undergo glycolysis. In agreement with the in vitro data, upon development glcS(-) stalk cells died but most were not vacuolated. We conclude that a UDP glucose derivative (such as glycogen or maltose) plays an essential energy independent role in autophagic cell death. PMID- 18424910 TI - Silencing of Bcl-2 expression by small interfering RNA induces autophagic cell death in MCF-7 breast cancer cells. AB - Apoptosis (programmed cell death type I) and autophagy (type II) are crucial mechanisms regulating cell death and homeostasis. The Bcl-2 proto-oncogene is overexpressed in 50-70% of breast cancers, potentially leading to resistance to chemotherapy, radiation and hormone therapy-induced apoptosis. Here, we investigated the role of Bcl-2 in autophagy in breast cancer cells. Silencing of Bcl-2 by siRNA in MCF-7 breast cancer cells downregulated Bcl-2 protein levels (>85%) and led to inhibition of cell growth (71%) colony formation (79%), and cell death (up to 55%) by autophagy but not apoptosis. Induction of autophagy was demonstrated by acridine orange staining, electron microscopy and an accumulation of GFP-LC3-II in autophagosomal membranes in MCF-7 cells transfected with GFP-LC 3(GFP-ATG8). Silencing of Bcl-2 by siRNA also led to induction of LC-3-II, a hallmark of autophagy, ATG5 and Beclin-1 autophagy promoting proteins. Knockdown of ATG5 significantly inhibited Bcl-2 siRNA-induced LC3-II expression, the number of GFP-LC3-II-labeled autophagosome positive cells and autophagic cell death (p < 0.05). Furthermore, doxorubicin at a high dose (IC(95), 1 microM) induced apoptosis but at a low dose (IC(50), 0.07 microM) induced only autophagy and Beclin-1 expression. When combined with Bcl-2 siRNA, doxorubicin (IC(50)) enhanced autophagy as indicated by the increased number cells with GFP-LC3-II stained autophagosomes (punctuated pattern positive). These results provided the first evidence that targeted silencing of Bcl-2 induces autophagic cell death in MCF-7 breast cancer cells and that Bcl-2 siRNA may be used as a therapeutic strategy alone or in combination with chemotherapy in breast cancer cells that overexpress Bcl-2. PMID- 18424911 TI - Loss of Pten, a tumor suppressor, causes the strong inhibition of autophagy without affecting LC3 lipidation. AB - (1)Pten (phosphatase and tensin homolog deleted on chromosome ten), a tumor suppressor, is a phosphatase with a variety of substrate specificities. Its function as a negative regulator of the class I phosphatidyl-inositol 3 kinase/Akt pathway antagonizes insulin-dependent cell signaling. The targeted deletion of Pten in mouse liver leads to insulin hypersensitivity and the upregulation of the phosphatidyl-inositol 3-kinase/Akt signaling pathway. In this study, we investigated the effects of Pten deficiency on autophagy, a major cellular degradative system responsible for the turnover of cell constituents. The autophagic degradation of [(14)C-leucine-labeled proteins of hepatocytes isolated from Pten-deficient livers was strongly inhibited, compared with that of control hepatocytes. However, no significant difference was found in the levels of the Atg12-Atg5 conjugate and LC3-II, the lipidated form of LC3, an intrinsic autophagosomal membrane marker, between control and Pten-deficient livers. Electron microscopic analyses showed that numerous autophagic vacuoles (autophagosomes plus autolysosomes) were present in the livers of control mice that had been starved for 48 hours, whereas they were markedly reduced in Pten deficient livers under the same conditions. In vivo administration of leupeptin to control livers caused the inhibition of autophagic proteolysis, resulting in the accumulation of autolysosomes. These autolysosomes could be separated as a denser autolysosomal fraction from other cell membranes by Percoll density gradient centrifugation. In leupeptin-administered mutant livers, however, the accumulation of denser autolysosomes was reduced substantially. Collectively, we conclude that enhanced insulin signaling in Pten deficiency suppresses autophagy at the formation and maturation steps of autophagosomes, without inhibiting ATG conjugation reactions. PMID- 18424913 TI - Rottlerin induces autophagy and apoptotic cell death through a PKC-delta independent pathway in HT1080 human fibrosarcoma cells: the protective role of autophagy in apoptosis. AB - Rottlerin is widely used as a protein kinase C-delta inhibitor. Recently, several reports have shown the possible apoptosis-inducing effect of rottlerin in some cancer cell lines. Here we report that rottlerin induces not only apoptosis but also autophagy via a PKC-delta-independent pathway in HT1080 human fibrosarcoma cells. Rottlerin treatment induced a dose- and time-dependent inhibition of cell growth, and cytoplasmic vacuolations were markedly shown. These vacuoles were identified as acidic autolysosomes by electron microscopy, acidic vesicular organelle (AVO) staining and transfection of green fluorescent protein-LC3. The LC3-II protein level also increased after treatment with rottlerin. Prolonged exposure to rottlerin eventually caused apoptosis via loss of mitochondrial membrane potential and translocation of AIF from mitochondria to the nucleus. However, the activities of caspase-3, -8 and -9 were not changed, and PARP did not show signs of cleavage. Interestingly, the pretreatment of cells with a specific inhibitor of autophagy (3-methyladenine) accelerated rottlerin-induced apoptosis as revealed by an analysis of the subdiploid fraction and TUNEL assay. Nevertheless, the knockdown of PKC-delta by RNA interference neither affected cell growth nor acidic vacuole formation. Similarly, rottlerin-induced cell death was not prevented by PKC-delta overexpression. Taken together, these findings suggest that rottlerin induces early autophagy and late apoptosis in a PKC-delta independent manner, and the rottlerin-induced early autophagy may act as a survival mechanism against late apoptosis in HT1080 human fibrosarcoma cells. PMID- 18424912 TI - Autophagy upregulation by inhibitors of caspase-3 and mTOR enhances radiotherapy in a mouse model of lung cancer. AB - Autophagy has been reported to be increased in irradiated cancer cells resistant to various apoptotic stimuli. We therefore hypothesized that induction of autophagy via mTOR inhibition could enhance radiosensitization in apoptosis inhibited H460 lung cancer cells in vitro and in a lung cancer xenograft model. To test this hypothesis, combinations of Z-DEVD (caspase-3 inhibitor), RAD001 (mTOR inhibitor) and irradiation were tested in cell and mouse models. The combination of Z-DEVD and RAD001 more potently radiosensitized H460 cells than individual treatment alone. The enhancement in radiation response was not only evident in clonogenic survival assays, but also was demonstrated through markedly reduced tumor growth, cellular proliferation (Ki67 staining), apoptosis (TUNEL staining) and angiogenesis (vWF staining) in vivo. Additionally, upregulation of autophagy as measured by increased GFP-LC3-tagged autophagosome formation accompanied the noted radiosensitization in vitro and in vivo. The greatest induction of autophagy and associated radiation toxicity was exhibited in the tri modality treatment group. Autophagy marker, LC-3-II, was reduced by 3 methyladenine (3-MA), a known inhibitor of autophagy, but further increased by the addition of lysosomal protease inhibitors (pepstatin A and E64d), demonstrating that there is autophagic induction through type III PI3 kinase during the combined therapy. Knocking down of ATG5 and beclin-1, two essential autophagic molecules, resulted in radiation resistance of lung cancer cells. Our report suggests that combined inhibition of apoptosis and mTOR during radiotherapy is a potential therapeutic strategy to enhance radiation therapy in patients with non-small cell lung cancer. PMID- 18424914 TI - Potentiation of the antitumor effects of both selective cyclooxygenase-1 and cyclooxygenase-2 inhibitors in human hepatic cancer cells by inhibition of the MEK/ERK pathway. AB - The molecular mechanisms behind the anti-neoplastic effects of non-steroidal anti inflammatory drugs (NSAIDs) are not completely understood and cannot be explained by the inhibition of the cyclooxygenase (COX) enzymes COX-1 and COX-2 alone. We previously reported that both the selective COX-1 inhibitor SC-560 and the selective COX-2 inhibitor CAY10404 exhibit anti-tumor effects in human hepatoma cells. NSAID inhibitors have many COX-independent actions and, among others, the mitogen-activated protein kinase (MAPK) pathways are targets for NSAIDs. Here, we examined the role of MEK/ERK1/2 signaling in the anti-neoplastic effects of both selective COX-1 and COX-2 inhibitors in two human hepatoma cell lines. Treatment of hepatoma cells with the selective COX-1 inhibitor SC-560, as well as with the selective COX-2 inhibitor CAY10404, was associated with activation of ERK1/2 in a time- and dose-dependent manner. Treatment with COX-1 and COX-2 inhibitors in the presence of the selective MEK1/2 inhibitor U0126 effectively suppressed ERK1/2 activation and combinations of either SC-560 or CAY10404 with U0126 resulted in synergistic effects on cell growth inhibition and induction of apoptosis. In HuH 6 hepatoma cells the combination-induced apoptosis was associated with caspase-9 and -3 activation, PARP cleavage, release of cytochrome c from the mitochondria into the cytosol and down-regulation of survivin and beta-catenin levels. In conclusion, our study showed that growth inhibitory concentrations of selective COX-1 and COX-2 inhibitors increased ERK1/2 phosphorylation in hepatoma cells, and that inhibition of the MEK/ERK signaling pathway potentiates the antitumor activity of both types of inhibitors. Therefore, our results provide preclinical support for a combined chemotherapeutic approach with selective NSAIDs and MEK inhibitors for the treatment of hepatocellular carcinoma. PMID- 18424915 TI - DNA replication, cell cycle progression and the targeted gene repair reaction. AB - Single-stranded oligonucleotides (ssODNs) can direct base changes in mammalian cells and influence changes in phenotype. The mechanism by which ssODNs alters the sequence is being revealed by studies carried out in model systems. In the long run, this information will provide the basis for clinical protocols designed to target genetic diseases. It is now clear that DNA replication plays an important part in the gene repair reaction. Here, we examine gene repair as a function of the amount of cells passing through S phase. We find that cells in mid to late S are most amenable to gene repair, and reaction manipulations that enrich the population of cells in S phase naturally lead to elevated correction frequencies. Our data suggest that these intra-S sub phases support higher levels of repair independent of transfection efficiencies or the rates of replication. A preliminary gene expression profile of cells in the most amenable correction phase indicates that the levels of cyclin G(2), cyclin H, CDK12A and CDK12B are raised significantly. Taken together, our data identify sections of S phase that enable higher levels of gene repair and establish a mechanistic framework for the use of gene repair in clinical setup. PMID- 18424916 TI - Bad things happen in the basal layer: KLF4 and squamous cell carcinoma. PMID- 18424917 TI - Enhanced EGFR inhibition and distinct epitope recognition by EGFR antagonistic mAbs C225 and 425. AB - Monoclonal antibodies (mAbs) that inhibit activation of the epidermal growth factor receptor (EGFR) have shown therapeutic potential in select malignancies including breast cancer. Here, we describe that combined use of two such mAbs, C225 (Cetuximab) and 425 (EMD55900), reduced growth and survival of EGFR overexpressing MDA-MB-468 breast cancer cells more effectively than either antibody alone. Similarly, the C225/425 antibody combination more effectively inhibited AKT and MAPK phosphorylation in MDA-MB-468 cells. Surface plasmon resonance, size exclusion chromatography and analytical ultracentrifugation demonstrated that mAbs C225 and 425 simultaneously bind to distinct antigenic epitopes on domain III of the soluble wild-type EGFR. Furthermore, neither mAb competed with the other for binding to cells expressing either wild-type EGFR or a mutant EGFR (EGFRvIII) associated with neoplasia. Mutagenesis experiments revealed that residues S460/G461 in EGFR domain III are essential components of the 425 epitope and clearly distinguish it from the EGF/ TGFalpha binding site and the C225 interaction interface. Collectively, these results support the conclusion that therapeutic EGFR blockade in cancer patients by combined use of mAbs C225 and 425 could provide advantages over the use of the two antibodies as single agents. PMID- 18424918 TI - Vaccine knowledge and practices of primary care providers of exempt vs. vaccinated children. AB - OBJECTIVES: Compare vaccine knowledge, attitudes and practices of primary care providers for fully vaccinated children and children who are exempt from school immunization requirements. METHODS: We conducted a mailed survey of parent identified primary care providers from four states to measure perceived risks and benefits of vaccination and other key immunization beliefs. Frequencies of responses were stratified by type of provider, identified by exempt versus vaccinated children. Logistic regression was used to calculate odds ratios for responses by provider type. RESULTS: 551 surveys were completed (84.3% response rate). Providers for exempt children had similar attitudes to providers for non exempt children. However, there were statistically significant increased concerns among providers for exempt children regarding vaccine safety and lack of perceived individual and community benefits for vaccines compared to other providers. CONCLUSIONS: The great majority of providers for exempt children had similar attitudes about vaccine safety, effectiveness and benefits as providers of non-exempt children. Although providers for exempt children were more likely to believe that multiple vaccines weaken a child's immune system and were concerned about vaccine safety and less likely to consider vaccines were beneficial, a substantial proportion of providers of both exempt and vaccinated children have concerns about vaccine safety and believe that CDC underestimates the frequency of vaccine side effects. Effective continuing education of providers about the risks and benefits of immunization and including in vaccine recommendations more information on pre and post licensing vaccine safety evaluations may help address these concerns. PMID- 18424919 TI - Autophagy: an overlooked mechanism of HIV-1 pathogenesis and neuroAIDS? AB - Human immunodeficiency virus type 1 (HIV-1) establishes a persistent infection characterized by progressive depletion of CD4(+) lymphocytes and immunosuppression. Although extensive research has examined the importance of apoptosis as a cause of cell death associated with HIV-1 infection, the role of autophagy has been largely ignored. Our laboratory has examined the autophagic process in HIV-1-infected cells. Following infection of human peripheral blood CD4(+) T-cells or U937 cells with HIV-1 for 48 hours, the autophagy proteins Beclin 1 and LC3-II were found to be markedly decreased. Beclin 1 mRNA expression and autophagosomes were also reduced in HIV-1 infected cells. Thus, our data indicate that HIV-1 infection inhibits autophagy in infected cells in contrast to the previously described induction of autophagy by gp120 in uninfected bystander cells. It is likely that HIV-1 has evolved this mechanism as part of an elaborate attempt to evade the immune system while promoting its own replication. We believe that autophagy is an overlooked mechanism in HIV-1 pathogenesis and plays a particularly important role in the early cognitive impairment and dementia often associated with advanced AIDS. A model is presented that describes the potential role of autophagy in NeuroAIDS. PMID- 18424947 TI - Manganese enhanced magnetic resonance imaging in a contusion model of spinal cord injury in rats: correlation with motor function. AB - OBJECTIVES: Various models of spinal cord injury in rodents have been established, and also techniques for lesion quantification. Measurement of the extent of the underlying injury is essential for monitoring the reproducibility of the experimental injury and assessment of therapeutic effects. In this study, we tested manganese-enhanced magnetic resonance imaging (MEMRI) for postmortem quantification of experimental spinal cord injury in rats. MATERIALS AND METHODS: Twelve rats were subjected to contusion injuries at the 11th thoracic vertebra, followed by MnCl2 injections into the cisterna magna. After 3 days of observation, postmortem MEMRI-features were correlated with values of locomotion testing and histology. RESULTS: MnCl2 yielded a strong contrast enhancement of the uninjured spinal cord, whereas no enhancement was observed at the injury site or caudally. Magnetic resonance imaging findings correlate closely with locomotor ratings. CONCLUSIONS: MEMRI represents a reliable method for visualization and functional assessment of spinal cord integrity in rats. PMID- 18424948 TI - Left ventricular and left atrial dimensions and volumes: comparison between dual source CT and echocardiography. AB - OBJECTIVES: We sought to determine the agreement for the quantification of cardiac chamber dimensions, volumes, and myocardial mass between dual-source computed tomography (DSCT) and echocardiography. MATERIAL AND METHODS: One hundred patients underwent DSCT and transthoracal echocardiography within 1 week. Measurements of dimensions were obtained in standardized planes in end-systole and end-diastole and included the anterior-posterior diameter of the left atrium, septal and posterior wall thickness, and inner diameter of the left ventricle. Global left ventricular (LV) functional parameters [end-systolic volume (ESV), end-diastolic volume (EDV), ejection fraction, and LV myocardial mass (LVMM)] were computed using semiautomated software. ESV, EDV, and LVMM were normalized to the body-surface-area (BSA). Intraobserver and interobserver agreement of DSCT analysis was assessed. Correlation between DSCT and echocardiography was tested through linear regression and Bland-Altman analysis. RESULTS: DSCT measurements had an excellent inter- and intraobserver agreement with close limits of agreement (R = 0.85-0.99, P < 0.001). All measurements obtained with DSCT showed a significant correlation with echocardiography, with close limits of agreement between modalities for all parameters. Significant differences of the mean difference from zero were only found for septal and posterior wall thickness (P < 0.001) (with a homogenous underestimation) and for EDV/BSA (P < 0.05) (showing an overestimation) in DSCT compared with echocardiography. No significant directional measurement bias was found for any parameter except for LVMM/BSA (R = 0.24, P < 0.05). CONCLUSION: Our results indicate that DSCT provides reliable measurements of LV dimensions, volumes, and myocardial mass with similar values as compared with echocardiography. PMID- 18424949 TI - Positron emission tomography/computed tomography and whole-body magnetic resonance imaging in staging of advanced nonsmall cell lung cancer--initial results. AB - OBJECTIVE: To evaluate and compare positron emission tomography/computed tomography (PET/CT) with whole-body magnetic resonance imaging (wbMRI) in the correct staging of patients with advanced nonsmall cell lung cancer (NSCLC). MATERIALS AND METHODS: Fifty-two patients with an NSCLC stage IIIa or IIIb (36 males and 16 females) were included in this study. Patients were referred to our department for restaging. Within 1 week PET/CT and wbMRI were performed in all patients. Images were examined independently by 2 experienced physicians from the Department of Nuclear Medicine and Radiology. Afterward, consensus reading was performed. In 22 patients, surgery served as gold standard, whereas in 30 patients, follow-up controls (after 2 months) were performed. RESULTS: The use of wbMRI correctly T-staged all patients. Especially volume interpolated breathhold examination sequence correctly T-staged all tumors. PET/CT did not correctly stage chest wall infiltration in 4 cases [sensitivity 92.3% (P < 0.05 to wbMRI)/specificity 100%], verified by surgery. PET/CT correctly N-staged 51 patients (sensitivity 96.1%/specificity 100%). WbMRI showed a significant tendency to understage N-status [sensitivity 88.5% (P < 0.05)/specificity 96.1%]. Different N-status by PET/CT changed operability in 4 patients. In 2 patients, distant metastases were detected by both techniques. CONCLUSION: In the correct staging of advanced NSCLC, PET/CT has advantages in N-staging. This is of high relevance for therapy planning. WbMRI especially using volume interpolated breathhold examination sequences, has certain advantages in T-staging. PMID- 18424950 TI - Magnetic resonance imaging assays for dimethyl sulfoxide effect on cancer vasculature. AB - OBJECTIVES: To evaluate the potential of quantitative assays of vascular characteristics based on dynamic contrast-enhanced magnetic resonance imaging (MRI) using a macromolecular contrast medium (MMCM) to search for and measure effects of dimethyl sulfoxide (DMSO) on cancer vasculature with microscopic correlations. MATERIAL AND METHODS: Saline-treated control (n = 8) and DMSO treated (n = 7) human breast cancer xenografts (MDA-MB-435) in rats were imaged dynamically by MMCM-enhanced MRI using albumin-(Gd-DTPA)27-(biotin)11 (molecular weight approximately 90 kDa), before and after a 1-week, 3-dose treatment course. After the posttreatment MRI examinations, tumors were perfused with lectin and fixative and subsequently stained with RECA-1 and streptavidin for quantitative fluorescent microscopy. Quantitative MRI estimates of cancer microvessel permeability (KPS; microL/min.100 cm3) and fractional plasma volume (fPV; %) were based on a 2-compartment kinetic model. Fluorescent microscopy yielded estimates of MMCM extravasation and vascular density that were compared to the MRI results. RESULTS: DMSO decreased cancer vascular endothelial permeability significantly (P < 0.05) from tumor KPSday0 = 19.3 +/- 8.8 microL/min.100 cm3 to KPSday7 = 0 microL/min.100 cm3). K values in the saline-treated tumors did not change significantly. The amount of extravasated albumin-Gd-(DTPA)27-(biotin)11, as assayed by a fluorescently labeled streptavidin stain that strongly binds to the biotin tag on the MMCM, was significantly (P < 0.05) lower in the DMSO-treated cancers than in the control cancers (57.7% +/- 5.5% vs. 34.2% +/- 4.9%). Tumor vascular richness as reflected by the MRI-assayed fPV and by the RECA-1 and lectin-stained microscopy did not change significantly with DMSO or saline treatment. CONCLUSION: Reductions in cancer microvascular leakiness induced by a 7-day course of DMSO could be detected and measured by dynamic MMCM-enhanced MRI and were confirmed by microscopic measurements of the leaked macromolecular agents in the same cancers. Results support the robustness of an MMCM-enhanced MRI approach to the characterization of cancers and providing first evidence for an in vivo effect of DMSO on cancer blood vessels. PMID- 18424951 TI - Noncontrast 3D steady state free precession magnetic resonance angiography of the thoracic central veins using nonselective radiofrequency excitation over a large field of view: initial experience. AB - PURPOSE: To evaluate the feasibility of three-dimensional (3D) steady state free precession (SSFP) magnetic resonance angiography (MRA) using nonselective radiofrequency excitation for the assessment of thoracic central veins. MATERIALS AND METHODS: Thirty consecutive patients (17 males, 13 females, age range 22-76) with various cardiac and thoracic vascular diseases underwent free-breathing electrocardiogram-gated noncontrast SSFP MRA and conventional high-resolution 3D contrast-enhanced (CE) MRA of the thorax at 1.5 T. Two readers evaluated both datasets for findings: venous visibility and sharpness (from 0, not visualized to 3, excellent definition); artifacts; signal-to-noise ratio (SNR); and contrast-to noise ratio (CNR) in 8 venous segments including superior vena cava (SVC), supra diaphragmatic inferior vena cava, bilateral brachiocephalic, proximal subclavian, and lower internal jugular veins. Statistical analysis was performed using Wilcoxon test for overall image quality and vessel visibility, t test for SNR and CNR analysis, and kappa coefficient for inter-observer variability. RESULTS: 3D SSFP and CE-MRA were successfully performed in all patients. Scan time for SSFP MRA ranged from 5 to 10 minutes (mean +/- standard deviation, 7 +/- 2 minutes). Reader 1 (2) graded the overall image quality as excellent and good on SSFP MRA in 23 (25) and 7 (5) patients, and on CE-MRA in 22 (23) and 8 (9) patients, respectively. On SSFP MRA, readers 1 and 2 graded 234 (97.5%) and 233 (97.1%) venous segments with diagnostic definition (grades 2 and 3) (kappa = 0.69), respectively. On conventional CE-MRA, readers 1 and 2 graded 231 (96.3%) and 232 (96.7%) venous segments with diagnostic definition (grades 2 and 3) (kappa = 0.68), respectively. Segmental visibility and sharpness were higher for lower internal jugular veins on CE-MRA for each reader (P < 0.001). No significant difference existed for venous visibility and sharpness scores for other venous segments between the 2 techniques for both readers (P > 0.05). SNR and CNR values were lower for internal jugular veins on SSFP MRA (P < 0.001). No significant difference existed between SNR and CNR values for the other venous segments on SSFP and CE-MRA (P > 0.05 for all). The 2 readers demonstrated patent SVC Glenn shunt to main pulmonary artery (n = 3), patent extra cardiac Fontan shunt from inferior vena cava to pulmonary artery confluence (n = 2), and dilatation and thrombosis of SVC (n = 1) and right brachiocephalic vein (n = 1) on both datasets. CONCLUSION: Free breathing navigator-gated noncontrast 3D SSFP MRA with nonselective radiofrequency excitation provides high image quality and sufficient SNR and CNR for confident evaluation of thoracic central veins. PMID- 18424952 TI - Quantification of coronary plaque by 64-slice computed tomography: a comparison with quantitative intracoronary ultrasound. AB - BACKGROUND: Noninvasive assessment of coronary atherosclerotic plaque may be useful for risk stratification and treatment of atherosclerosis. MATERIALS AND METHODS: We studied 47 patients to investigate the accuracy of coronary plaque volume measurement acquired with 64-slice multislice computed tomography (MSCT), using newly developed quantification software, when compared with quantitative intracoronary ultrasound (QCU). Quantitative MSCT coronary angiography (QMSCT-CA) was performed to determine plaque volume for a matched region of interest (regional plaque burden) and in significant plaque defined as a plaque with > or =50% area obstruction in QCU, and compared with QCU. Dataset with image blurring and heavy calcification were excluded from analysis. RESULTS: In 100 comparable regions of interest, regional plaque burden was highly correlated (coefficient r = 0.96; P < 0.001) between QCU and QMSCT-CA, but QMSCT-CA overestimated the plaque burden by a mean difference of 7 +/- 33 mm3 (P = 0.03). In 76 significant plaques detected within the regions of interest, plaque volume determined by QMSCT-CA was highly correlated (r = 0.98; P < 0.001) with a slight underestimation of 2 +/- 17 mm3 (P = not significant) when compared with QCU. Calcified and mixed plaque volume was slightly overestimated by 4 +/- 19 mm3 (P = ns) and noncalcified plaque volume was significantly underestimated by 9 +/- 11 mm3 (P < 0.001) with QMSCT-CA. Overall, the limits of agreement for plaque burden/volume measurement between QCU and QMSCT-CA were relatively large. Reproducibility for the measurements of regional plaque burden with QMSCT-CA was good, with a mean intraobserver and interobserver variability of 0% +/- 16% and 4% +/- 24%, respectively. CONCLUSIONS: Quantification of coronary plaque within selected proximal or middle coronary segments without image blurring and heavy calcification with 64-slice CT was moderately accurate with respect to intravascular ultrasound and demonstrated good reproducibility. Further improvement in CT resolution is required for more reliable measurement of coronary plaques using quantification software. PMID- 18424953 TI - Acoustic attenuation by contrast agent microbubbles in superficial tissue markedly diminishes petechiae bioeffects in deep tissue. AB - OBJECTIVE: To measure how ultrasound attenuation by contrast agent microbubbles (MBs) in superficial tissue affects petechiae creation in underlying deep tissue. MATERIALS AND METHODS: Studies using Sprague-Dawley rats were approved by the Animal Care and Use Committee. MBs were injected intravenously, and 12 ultrasound pulses (100 sinusoids of 1 MHz ultrasound per pulse) were applied through the skin overlying the hindlimb adductors at intervals of 10 or 60 seconds. In some groups, the skin was resected and immediately returned without re-establishing vascular connections. Muscle petechiae were counted. RESULTS: Applying ultrasound through unperfused skin after bolus and continuous intravenous MB injection yielded, respectively, 30-fold and 3.5-fold more petechiae than for perfused skin. Surprisingly, petechiae/mm2 decreased with a higher MB dosage [0.12 +/- 0.05 (1 x 10 MBs/g) vs. 0.04 +/- 0.02 (3 x 10 MBs/g)] when ultrasound was applied through perfused skin. In contrast, petechiae/mm2 was approximately proportional to MB dosage for unperfused skin [0.17 +/- 0.10(5) (1 x 10 MBs/g) vs. 0.42 + 0.14 (3 x 10(5) MBs/g)]. In comparison to MB-free controls, MB solutions in this concentration range reduced the peak-negative pressure of ultrasound by 65% to 85%. CONCLUSIONS: Acoustic attenuation by MBs in skin markedly reduces petechiae creation in deep muscle. Petechiae inhibition is dependent on [MB]2.1 and, therefore, dominates the otherwise proportional relationship between petechiae and [MB] in muscle. The drop of peak-negative pressure below a critical microvessel rupturing threshold is the probable mechanism for petechiae inhibition. These results indicate that high MB doses could, paradoxically, reduce the potential for petechiae creation and may have important bearing on the design of contrast ultrasound-based therapeutics. PMID- 18424954 TI - Quantification of pancreatic lipomatosis and liver steatosis by MRI: comparison of in/opposed-phase and spectral-spatial excitation techniques. AB - OBJECTIVES: The goal of the present study was the assessment of pancreatic and hepatic fat content applying 2 established magnetic resonance (MR) imaging techniques: in-phase/opposed-phase gradient-echo MR imaging and fat-selective spectral-spatial gradient-echo imaging. Results of both approaches were compared, and influences of T1- and T2*-related corrections were assessed. The possibility of a correlation between pancreatic lipomatosis and liver steatosis was investigated. MATERIALS AND METHODS: Seventeen volunteers at risk for type 2 diabetes (6 male, 11 female; age, 26-70 years; body mass index, 19.4-41.3 kg/m2; mean, 31.7 kg/m2) were examined. Liver and pancreas fat content were quantified with 2 different gradient-echo techniques: one uses a spectral-spatial excitation technique with 6 binomial radio frequency pulses, which combines chemical shift selectivity with simultaneous slice-selective excitation. The other technique based on double-echo chemical shift gradient-echo MR provides in- and opposed phase images simultaneously. Influences of T1 and individual T2* effects on results using in-phase/opposed-phase imaging were estimated and corrected for, based on additional T2* measurements. RESULTS: The fat content calculated from images recorded with the fat-selective spectral-spatial gradient-echo sequence correlated well with the fat fraction determined with in-phase/opposed-phase imaging and following correction for T1/T2* effects: pancreas r = 0.93 (P < 0.0001) and liver r = 0.96 (P < 0.0001). In-phase/opposed-phase imaging revealed a pancreatic fat content between 1.6% and 22.2% (mean, 8.8% +/- 5.7%) and a hepatic fat fraction between 0.6% and 33.3% (mean, 7.9% +/- 9.1%). The fat selective spectral-spatial gradient-echo sequence revealed a pancreatic lipid content between 3.4% and 16.1% (mean, 9.8% +/- 4.0%) and a hepatic fat content between 0% and 28.5% (mean, 8.8% +/- 8.3%). With neither technique was a substantial correlation between pancreatic and hepatic fat content found. CONCLUSION: The presented results suggest that both methods are reliable tools for pancreatic and hepatic fat quantification. However, for reliable assessment of quantitative fat by the in-phase/opposed-phase technique, an additional measurement of T2* seems crucial. PMID- 18424956 TI - Validity of the animal models for nephrogenic systemic fibrosis. PMID- 18424957 TI - An alternative source of autograft bone for spinal fusion: the femur: technical case report. AB - OBJECTIVE: Autograft bone obtained from the iliac crest remains the "gold standard" for spinal fusion. For various reasons, including previous harvesting or pelvic dysmorphism, the iliac crest bone graft may not be available to the spinal surgeon. We present a novel use of a common orthopedic procedure, intramedullary reaming, for obtaining autograft for revision spinal fusion. METHODS: A 47-year-old woman presented with failed back syndrome after multiple lumbar surgeries with previous bilateral iliac crest bone harvest. A commercially available reaming system (Synthes Reamer-Irrigator-Aspirator; Synthes USA, West Chester, PA) was introduced into the left intramedullary canal of the femur while the patient remained in the prone position. Using continuous irrigation and aspiration, the reaming debris was collected and used as autograft for the subsequent spinal fusion. RESULTS: The patient underwent a successful L4-L5, L5 S1 transforaminal lumbar interbody fusion with L3-S1 pedicle screw fixation. No complications from the femoral reaming were observed, and 6-month follow-up x rays demonstrated osseous fusion. CONCLUSION: Femoral reaming provides an alternative source of autograft bone when other sources are unavailable. PMID- 18424961 TI - Microsurgical anatomy of the supracerebellar transtentorial approach to the posterior mediobasal temporal region: technical considerations with a case illustration. AB - OBJECTIVE: Surgical access to the posterior portion of the mediobasal temporal lobe presents a formidable challenge to neurosurgeons, and much controversy still exists regarding the selection of the surgical approach to this region. The supracerebellar transtentorial (SCTT) approach to the posterior mediobasal temporal region can be used as an alternative to the subtemporal or transtemporal approaches. The aim of this study was to demonstrate the surgical anatomy of the SCTT approach and review the gyral, sulcal, and vascular anatomy of the posterior mediobasal temporal lobe. The use of this approach in the resection of a ganglioglioma located in the left posterior parahippocampal gyrus is illustrated. METHODS: The SCTT approach to the posterior parahippocampal gyrus was performed on three silicone-injected cadaveric heads. The gyral, sulcal, and arterial anatomy of the posterior mediobasal temporal lobe was studied in six formalin fixed injected hemispheres. RESULTS: The SCTT approach provided a direct path to the posterior mediobasal temporal lobe and exposed the posterior parahippocampal gyrus as well as the adjacent gyri in all of the cadaveric specimens. Through this approach, gross total resection of the ganglioglioma was possible in our patient. CONCLUSION: The SCTT approach provided a viable surgical route to the posterior mediobasal temporal lobe in the cadaveric studies. This approach provides an advantage over the subtemporal and transtemporal routes in that there is less temporal lobe retraction. PMID- 18424962 TI - Microsurgical anatomy of the safe entry zones on the anterolateral brainstem related to surgical approaches to cavernous malformations. AB - OBJECTIVE: To study the microanatomy of the brainstem related to the different safe entry zones used to approach intrinsic brainstem lesions. METHODS: Ten formalin-fixed and frozen brainstem specimens (20 sides) were analyzed. The white fiber dissection technique was used to study the intrinsic microsurgical anatomy as related to safe entry zones on the brainstem surface. Three anatomic landmarks on the anterolateral brainstem surface were selected: lateral mesencephalic sulcus, peritrigeminal area, and olivary body. Ten other specimens were used to study the axial sections of the inferior olivary nucleus. The clinical application of these anatomic nuances is presented. RESULTS: The lateral mesencephalic sulcus has a length of 7.4 to 13.3 mm (mean, 9.6 mm) and can be dissected safely in depths up to 4.9 to 11.7 mm (mean, 8.02 mm). In the peritrigeminal area, the distance of the fifth cranial nerve to the pyramidal tract is 3.1 to 5.7 mm (mean, 4.64 mm). The dissection may be performed 9.5 to 13.1 mm (mean, 11.2 mm) deeper, to the nucleus of the fifth cranial nerve. The inferior olivary nucleus provides safe access to lesions located up to 4.7 to 6.9 mm (mean, 5.52 mm) in the anterolateral aspect of the medulla. Clinical results confirm that these entry zones constitute surgical routes through which the brainstem may be safely approached. CONCLUSION: The white fiber dissection technique is a valuable tool for understanding the three-dimensional disposition of the anatomic structures. The lateral mesencephalic sulcus, the peritrigeminal area, and the inferior olivary nucleus provide surgical spaces and delineate the relatively safe alleys where the brainstem can be approached without injuring important neural structures. PMID- 18424963 TI - Image-guided lateral suboccipital approach: part 1-individualized landmarks for surgical planning. AB - OBJECTIVE: Being situated close to the transverse and sigmoid sinus, the asterion has traditionally been viewed as a landmark for surgical approaches to the posterior fossa. Cadaveric studies, however, have shown its variability in relation to underlying anatomic structures. We have used an image-guidance technology to determine the precise anatomic relationship between the asterion and the underlying transverse-sigmoid sinus transition (TST) complex in patients scheduled for posterior fossa surgery. The applicability of three-dimensional (3 D) volumetric image-rendering for presurgical anatomic identification and individualization of a surgical landmark was evaluated. METHODS: One-millimeter computed tomographic slices were combined with venous computed tomographic angiography in 100 patients, allowing for 3-D volumetric image-rendering of the cranial bone and the dural vasculature at the same time. The spatial relationship between the asterion and the TST was recorded bilaterally by using opacity modulation of the bony surface. The location of both the asterion and the TST could be confirmed during surgery in all of these patients. RESULTS: It was possible to accurately visualize the asterion and the sinuses in a single volumetrically rendered 3-D image in more than 90% of the patients. The variability in the anatomic position of the asterion as shown in cadaveric studies was confirmed, providing an individualized landmark for the patients. In this series, the asterion was located from 2 mm medial to 7 mm lateral and from 10 mm inferior to 17 mm superior to the TST, respectively. CONCLUSION: Volumetric image-rendering allows for precise in vivo measurements of anatomic distances in 3-D space. It is also a valuable tool for assessing the validity of traditional surgical landmarks and individualizing them for surgical planning. PMID- 18424964 TI - Image-guided lateral suboccipital approach: part 2-impact on complication rates and operation times. AB - OBJECTIVE: Image-guidance systems are widely available for surgical planning and intraoperative navigation. Recently, three-dimensional volumetric image rendering technology that increasingly applies in navigation systems to assist neurosurgical planning, e.g., for cranial base approaches. However, there is no systematic clinical study available that focuses on the impact of this image guidance technology on outcome parameters in suboccipital craniotomies. METHODS: A total of 200 patients with pathologies located in the cerebellopontine angle were reviewed, 100 of whom underwent volumetric neuronavigation and 100 of whom underwent treatment without intraoperative image guidance. This retrospective study analyzed the impact of image guidance on complication rates (venous sinus injury, venous air embolism, postoperative morbidity caused by venous air embolism) and operation times for the lateral suboccipital craniotomies performed with the patient in the semi-sitting position. RESULT: This study demonstrated a 4% incidence of injury to the transverse-sigmoid sinus complex in the image guided group compared with a 15% incidence in the non-image-guided group. Venous air embolisms were detected in 8% of the image-guided patients and in 19% of the non-image-guided patients. These differences in terms of complication rates were significant for both venous sinus injury and venous air embolism (P < 0.05). There was no difference in postoperative morbidity secondary to venous air embolism between both groups. The mean time for craniotomy was 21 minutes in the image-guided group and 39 minutes in non-image-guided group (P = 0.036). CONCLUSION: Volumetric image guidance provides fast and reliable three dimensional visualization of sinus anatomy in the posterior fossa, thereby significantly increasing speed and safety in lateral suboccipital approaches. PMID- 18424965 TI - Endoscopic sublabial transmaxillary approach to the rostral middle fossa. AB - OBJECTIVE: The rostral middle fossa faces the temporal pole and is the endocranial anterosuperior aspect of the greater wing of the sphenoid. Standard approaches to this region, such as the subtemporal, pterional, or orbitozygomatic approaches, require significant brain retraction or manipulation of the temporalis muscle. We report an endoscopic sublabial transmaxillary approach to this cranial base region that avoids the aforementioned pitfalls. METHODS: Ten adult cadaveric half heads were used to develop the endoscopic approach and to identify the salient surgical landmarks. RESULTS: The approach was divided into three stages: entry into the maxillary sinus, entry into the infratemporal fossa, and entry into the middle fossa. A craniotomy of greater than 20 mm in diameter can be safely created in the rostral middle fossa. When coupled with image guidance, the approach provides the flexibility to tailor the size and location of the middle fossa craniotomy. CONCLUSION: Although endonasal endoscopic approaches are increasing in popularity, the middle fossa has not been adequately accessed with these techniques. The endoscopic sublabial transmaxillary approach provides safe and direct access to the rostral middle fossa, eliminating the need for brain retraction, temporalis muscle manipulation, or an external incision. The approach also permits early devascularization of cranial- or dural-based lesions. PMID- 18424966 TI - Medial sphenoid ridge meningiomas: classification, microsurgical anatomy, operative nuances, and long-term surgical outcome in 35 consecutive patients. AB - OBJECTIVE: On the basis of contemporary multiplanar imaging, microsurgical observations, and long-term follow-up in 60 consecutive patients with sphenoid ridge meningiomas, we propose a modification to Cushing's classification of these tumors. This article will concentrate on patients from this series with global medial sphenoid ridge tumors. METHODS: Data were collected prospectively for 35 patients with global meningiomas arising from the medial portion of the sphenoid ridge that were surgically treated between 1982 and 2002. RESULTS: All patients were followed for the entire length of this study (mean, 12.8 yr). The tumor size ranged from 2 to 8 cm (mean, 4.5 cm). Of the 24 patients with purely intradural tumors, four (17%) had Simpson Grade I and 19 had Simpson Grade II resections; 23 (96%) had gross total resections. Of the 11 patients with tumors extending extradurally (i.e., cavernous sinus), one (9%) patient had a Simpson Grade II resection, whereas nine (82%) had Simpson Grade III resections, with the latter being all visible tumor removed except that in the cavernous sinus. One (9%) of these 11 patients had a gross total resection, and 9 (82%) had radical resections, with the latter defined as total removal of all intradural tumor. The overall morbidity rate was 18%. There was no surgical mortality or symptomatic cerebral infarction. CONCLUSION: An accurate classification of global medial sphenoid meningiomas is mandatory to gain insight into their clinical behavior and for understanding the long-term efficacy and safety of available treatment options. Primary medial sphenoid ridge tumors consistently involve the unilateral arteries of the anterior cerebral circulation, and therefore, the resection of tumor from around these arteries is the most important operative nuance for their safe excision. PMID- 18424967 TI - Purely endoscopic resection of colloid cysts. AB - OBJECTIVE: To further assess the safety and long-term efficacy of endoscopic resection of colloid cysts of the third ventricle. METHODS: A retrospective review of a series of 35 consecutive patients (18 male, 17 female) with colloid cysts treated by endoscopic surgery was undertaken. RESULTS: The mean patient age was 32.4 years (range, 11-54 yr). Headache was the most common presenting symptom (22 patients). The average tumor size was 18 mm (range, 3-50 mm). The endoscopic technique could not be completed in six patients, necessitating conversion to an open craniotomy and a transcortical approach to the colloid cyst. All patients had histologically confirmed colloid cysts of the third ventricle, and complete resection of the lesion was confirmed macroscopically and radiologically in all patients. There were no deaths. Two patients developed aseptic meningitis without any permanent sequelae. One patient developed unilateral hydrocephalus attributable to obstruction of the foramen of Monro, which was treated with endoscopic septum pellucidotomy. The median follow-up period was 88 months (range, 10-132 mo). There was one asymptomatic radiological recurrence. No seizures occurred after surgery. CONCLUSION: The results of this study support the role of endoscopic resection in the treatment of patients with colloid cysts as a safe and effective modality. In some cases, conversion to an open procedure may be required. Additional follow-up will be required to continue to address the duration of lesion-free survival. PMID- 18424968 TI - Endoscopic endonasal pituitary transposition for a transdorsum sellae approach to the interpeduncular cistern. AB - OBJECTIVE: The interpeduncular cistern, including the retroinfundibular area, is one of the most challenging regions to approach surgically. The pituitary gland and the infundibulum guard the region when an endonasal route is undertaken. Superior transposition of the pituitary gland and infundibulum is described as a functional means to access this complex region through a fully endoscopic, completely transnasal route. METHODS: Ten consecutive patients in whom a pituitary transposition was performed during an expanded endonasal approach at the University of Pittsburgh Medical Center for resection of retroinfundibular lesions were reviewed. The series consisted of seven men and three women with a mean age of 44.4 years. Pathology consisted of four craniopharyngiomas, four chordomas, and two petroclival meningiomas. RESULTS: Five patients (50%) underwent total resection of the tumor, three patients (30%) underwent near total resection ( > 95% removal), and two patients (20%) had partial resection of petroclival meningiomas with the goal of optic apparatus decompression. All four patients with visual deficits recovered their vision completely. There was no neurological deterioration. Eight patients had normal pituitary function preoperatively, seven of whom (87.5%) had confirmed function preservation postoperatively, with one of these patients experiencing transient diabetes insipidus. The remaining patient with a hypothalamic craniopharyngioma underwent complete resection with obligatory panhypopituitarism and diabetes insipidus. CONCLUSION: Endoscopic endonasal transposition of the pituitary gland and its stalk can provide a valuable corridor to the retroinfundibular space and interpeduncular cistern with pituitary function preservation in the majority of patients. This approach should only be pursued once significant experience with endoscopic endonasal approaches has been acquired. PMID- 18424969 TI - The juxtacondylar approach to the jugular foramen. AB - OBJECTIVE: We sought to describe the juxtacondylar approach to jugular foramen tumors. METHODS: Through an anterolateral approach, the third segment of the vertebral artery (between C2 and the dura mater) is controlled. The C1 transverse process of the atlas, which is located just inferiorly to the jugular foramen, is then removed. The dissection of the internal jugular vein is performed as high as possible, with control of the IXth, Xth, XIth, and XIIth cranial nerves. If required by a tumor extending into the neck, the internal and external carotid arteries can be exposed and controlled. Through a partial mastoidectomy and after removal of the bone covering the jugular tubercle, the end of the sigmoid sinus and then the posteroinferior part of the jugular foramen are reached. RESULTS: This technique is efficient to expose tumors extending into the jugular foramen. Contrary to the infratemporal approach, it has the main advantage of avoiding petrous bone drilling and associated potential complications. Lower cranial nerves are well exposed in the neck. In patients with schwannomas, complete resection with selective dividing of only the few involved rootlets can be achieved. CONCLUSION: The juxtacondylar approach is an efficient approach to tumors located in the jugular foramen. It necessitates control of the third segment of the vertebral artery but has the advantage of avoiding complications associated with petrous bone drilling. Extension beyond the jugular foramen requires combination with an infratemporal or a retrosigmoid approach. PMID- 18424970 TI - The new generation polestar n20 for conventional neurosurgical operating rooms: a preliminary report. AB - OBJECTIVE: The objective of this work is to present the preliminary clinical experience we acquired in using the new PoleStar generation, N20 (Medtronic Navigation, Louisville, CO), in a modified conventional operating room. METHODS: PoleStar N20 is a 0.15-T, intraoperative scanner combined with both an integrated optical and a magnetic resonance imaging tracking scanner. All standard imaging modes, such as T1, T2, and fluid-attenuated inversion recovery, are available through the magnet. To shield the operating room from radiofrequency interference, a Faraday cage was constructed using a conductive metal mesh installed under the wall decoration. Sixty-one patients, most of whom had gliomas or pituitary adenomas, underwent intraoperative magnetic resonance imaging in our clinic. The extent of resection and the surgical consequences of intraoperative imaging were analyzed. DISCUSSION: The image quality for T1-weighted, gadolinium enhanced tumors was sufficiently good to enable us to evaluate the extent of tumor resection, whereas the T2-weighted image quality must be improved. New technologies, such as high-temperature superconductive coils and ultra-small super-paramagnetic iron particles, e.g., ferumoxtran-10, can lead to a dramatic improvement in image quality, heralding the commencement of the widespread use of intraoperative magnetic resonance imaging. CONCLUSION: The acquisition of the PoleStar N20 opened new horizons in the treatment of our patients. This novel, compact, intraoperative magnetic resonance imaging scanner can be installed in a standard operating room without major modifications. Standard surgical instruments can be used. Intraoperative magnetic resonance imaging provided valuable information that allowed intraoperative modification of the surgical strategy. PMID- 18424971 TI - A comparative analysis of coregistered ultrasound and magnetic resonance imaging in neurosurgery. AB - OBJECTIVE: This work presents qualitative and quantitative side-by-side comparisons of oblique coregistered magnetic resonance imaging (MRI) scans and ultrasound images obtained during 35 neurosurgical procedures. METHODS: Spatially registered series of ultrasound images were recorded for subsequent off-line evaluation and comparison with corresponding preoperative MRI studies. The degree of misalignment was reduced by reregistering the target volume directly with segmented features. RESULTS: The initial apparent spatial misalignment of the target volume after craniotomy ranged from 0.11 to 8.73 mm (mean, 4.01 mm). After reregistration, the mutual information in overlapping segmented features was increased, presumably evidence of a better alignment locally. Additionally, the degree of feature congruence, which was assessed quantitatively through a convex hull approximation, demonstrated that the ultrasound volume was consistently smaller than its MRI counterpart. CONCLUSION: Although intraoperative ultrasound tends to be difficult to interpret by itself, when accurately coregistered with preoperative MRI scans, its potential utility as a navigational guide is enhanced. PMID- 18424972 TI - Usefulness of intraoperative photodynamic diagnosis using 5-aminolevulinic acid for meningiomas with cranial invasion: technical case report. AB - OBJECTIVE: We present a case of a meningioma in which photodynamic diagnosis (PDD) using 5-aminolevulinic acid was very useful in identifying the cranial involvement. CLINICAL PRESENTATION: An 83-year-old woman presented with a bony, hard, immobile bulge in her left forehead. Computed tomographic scans showed a thickening in the left frontal bone with a flat mass underneath. Magnetic resonance imaging scans revealed that enhancing lesions spread to the dura mater and subcutaneous tissue around the thickened frontal bone, reaching the upper margin of the left orbit. INTERVENTION: Intraoperative PDD using 5-aminolevulinic acid indicated the optimal extent of the excision by showing clear fluorescence of affected tissues. The tumor was totally resected and diagnosed as an atypical meningioma. Histopathological examination confirmed the consistency of the extent of tumor invasion with affected lesions on PDD. CONCLUSION: To the best of our knowledge, this is the first case demonstrating the efficacy of PDD using 5 aminolevulinic acid for a meningioma with cranial invasion. Additional studies are warranted, as shown in cases of malignant gliomas. PMID- 18424973 TI - Endoscope-controlled microneurosurgery to treat middle fossa epidermoid cysts: technical case report. AB - OBJECTIVE: To present an alternative technique of endoscope-controlled microneurosurgery for the treatment of middle fossa epidermoid cysts. METHODS: The three operations described were performed through an approximately 2-cm diameter temporal craniotomy after a straight skin incision was made. Resection was then performed under the magnification of a 30-degree rigid endoscope, which mandated the use of exclusively conventional microsurgical instruments. RESULTS: Total resection was accomplished in all three patients with large middle fossa epidermoid cysts through a small temporal corticectomy, without damage to neurovascular structures. CONCLUSION: This procedure allowed the association of a smaller craniotomy, better cosmetic results, and minor retraction of the brain to wide resection of the tumor and satisfactory functional outcomes. PMID- 18424974 TI - Endoscopic supracerebellar infratentorial approach for pineal cyst resection: technical case report. AB - OBJECTIVE: Accepted surgical strategies to address symptomatic pineal cysts include transventricular flexible or rigid endoscopy and supracerebellar infratentorial or occipital transtentorial microsurgical approaches. We report the first application of the endoscopic supracerebellar infratentorial approach for the complete resection of a pineal cyst. Unlike transventricular endoscopy, this technique poses no risk to the fornices and can be applied independent of ventricular size. CLINICAL PRESENTATION: A 37-year-old woman sought treatment for intractable headaches. A thorough evaluation revealed only a pineal cyst exerting mass effect on the tectum but causing no hydrocephalus. A period of nonoperative management was unsuccessful, and the patient was referred for surgery. TECHNIQUE: The patient was positioned in the semi-sitting position. The supracerebellar infratentorial corridor was accessed through a burr-hole. The pineal cyst was resected completely via the endoscope. Postoperatively, the patient's headaches resolved completely. CONCLUSION: The endoscopic supracerebellar infratentorial approach involves minimal brain retraction, poses no risk to the fornices, allows visualization and avoidance of the Galenic veins, and can be performed regardless of the size of the ventricle. Consequently, it is an excellent minimally invasive surgical option for resection or fenestration of symptomatic pineal cysts. PMID- 18424975 TI - Tentorial dural arteriovenous fistulae: operative strategies and microsurgical results for six types. AB - OBJECTIVE: Tentorial dural arteriovenous fistulae (DAVF) are rare, have a high risk of hemorrhage, often cannot be obliterated endovascularly, and frequently require microsurgical interruption of the draining vein. We differentiated these fistulae into six types and developed specific operative strategies on the basis of these types. METHODS: During a 9-year period, 31 patients underwent microsurgical treatment for tentorial fistulae: seven galenic DAVF, eight straight sinus DAVF, three torcular DAVF, three tentorial sinus DAVF, eight superior petrosal sinus DAVF, and two incisural DAVF. RESULTS: The posterior interhemispheric approach was used with galenic DAVF; the supracerebellar infratentorial approach was used with straight sinus DAVF; a torcular craniotomy was used with torcular DAVF; the supratentorial-infraoccipital approach was used with tentorial sinus DAVF; the extended retrosigmoid approach was used with superior petrosal sinus DAVF; and a pterional or subtemporal approach was used with incisural DAVF. Angiographically, 94% of the fistulae were obliterated completely. Four patients had transient neurological morbidity, none had permanent neurological morbidity; and there was no operative mortality (mean follow-up, 4.2 yr). CONCLUSION: Tentorial DAVF can be differentiated on the basis of fistula location, dural base, associated sinus, and direction of venous drainage. The operative strategy for each type is almost algorithmic, with each type having an optimum surgical approach and an optimum patient position that allows gravity to retract the brain, open subarachnoid planes, and shorten dissection times. No matter the type, the fistula is treated microsurgically by simple interruption of the draining vein. PMID- 18424976 TI - Usefulness of preoperative three-dimensional computed tomographic angiography with two-dimensional computed tomographic imaging for rupture point detection of middle cerebral artery aneurysms. AB - OBJECTIVE: We report the technique of three-dimensional computed tomographic (CT) angiography with a two-dimensional CT image aiding in the early operation of ruptured middle cerebral artery aneurysms. This combined image allows the prediction of the rupture point in the aneurysm and may reduce the risk of rupture during early clipping surgery. METHODS: The findings for 14 patients with 14 middle cerebral artery ruptured aneurysms who underwent subsequent early clipping were analyzed. The average aneurysm size was 8.5 mm, and there were two large and one giant aneurysms. CT examinations were performed by means of a multidetector CT scanner (Aquilion M16; Toshiba Medical Systems, Tokyo, Japan) and reconstructed with a workstation (ZIO M900 QUADRA; Amin Co., Ltd., Tokyo, Japan). We constructed an operating view through three-dimensional CT angiography for a lateral transsylvian approach with a two-dimensional CT image (nonshaded volume-rendering image), which was perpendicular to the direction of the surgical approach. Using this combined image, we predicted the rupture point of the aneurysm and successfully performed clipping surgery through a lateral transsylvian approach. Rupture points were confirmed at the time of surgery. Rupture points of 13 out of 14 aneurysms appeared as we expected, but one differed; all aneurysms were successfully clipped. Thirteen of the 14 patients could be clipped without rupture at surgery, but the remaining patient experienced rupture just after craniotomy. CONCLUSION: The combination of three dimensional CT angiography and two-dimensional CT images may help improve the surgical outcome by indicating aneurysmal rupture points, leading to the prevention of rupture. PMID- 18424977 TI - External carotid artery to middle cerebral artery bypass with the saphenous vein graft. AB - Patients with occlusive cerebrovascular disease who have failed maximal medical therapy, which consists of antiplatelet agents as well as maximizing modifiable risk factors such as blood pressure, cholesterol, smoking cessation, and obesity, and whose lesions are not amenable or have not responded to the more common vascular procedures (i.e., carotid endarterectomy or stenting) are considered candidates for an extracranial-intracranial bypass. Additionally, for a patient to be a candidate, he/she must have an adequate graft vessel. Typically, this vessel is the superficial temporal artery. However, oftentimes, the superficial temporal artery is an inadequate vessel or the patient requires a high-flow conduit. It is in these patients that use of the saphenous vein should be considered. In this report, we detail the technical aspects of performing an extracranial-intracranial bypass by using a saphenous vein graft. PMID- 18424978 TI - Improved image interpretation with combined superselective and standard angiography (double injection technique) during embolization of arteriovenous malformations. AB - OBJECTIVE: Interpretation of angioarchitecture during embolization of intracranial arteriovenous malformations (AVMs) is critical to optimizing results. We describe an adjunctive technique to aid in the interpretation of AVM embolization and improve safety. METHODS: In the past 100 consecutive patients who underwent AVM embolization by a single surgeon (RAM), each AVM nidus was selectively catheterized and microangiography was performed. After the microcatheter contrast exited the AVM, guiding catheter angiography was performed during the same digital run. The microangiogram was digitally superimposed on the guiding catheter angiogram to delineate important landmarks such as the nidus perimeter, draining veins, and microcatheter tip, which were then drawn on the digital subtraction angiographic monitor with a marking pen in two orthogonal views. RESULTS: Important landmarks were continually visualized during the embolization procedure despite subtracted fluoroscopy ("blank" roadmap). These techniques qualitatively helped to: 1) appreciate the overall size and morphology of the nidus, 2) clearly visualize the safe limits of the embolic injection within the nidus perimeter, 3) clearly visualize draining patterns to help avoid premature venous embolization, 4) decipher small draining veins from arteries, 5) continuously monitor the location and status of the microcatheter tip, and 6) increase the confidence of the surgeon during prolonged embolic injections. CONCLUSION: The double injection technique, with marking pen demarcation of the nidus perimeter, venous drainage, and microcatheter tip position, was qualitatively useful in every case. PMID- 18424979 TI - Combined endoscope-assisted transclival clipping and endovascular stenting of a basilar trunk aneurysm: case report. AB - OBJECTIVE: We describe a patient with a mid-basilar aneurysm treated with combined endoscope-assisted transsphenoidal clipping and endovascular stenting. CLINICAL PRESENTATION: A 28-year-old woman was transferred to the cranial base surgery center with an acute Grade III subarachnoid hemorrhage. Cerebral angiography demonstrated a small basilar trunk aneurysm that was not amenable to acute endovascular treatment. INTERVENTION: The patient underwent sublabial transsphenoidal/transclival endoscope-assisted clipping of the aneurysm and subsequent stenting of the affected segment. The aneurysm was repaired with a low profile Weck clip (Weck Closure Systems Research, Triangle Park, NC) that permitted a watertight closure of the clival dura using cardiac Medtronic U-clips (Medtronic, Inc., Minneapolis, MN). Postoperatively, the patient had no evidence of cerebrospinal fluid leakage. CONCLUSION: Watertight dural closure was possible due to the use of a low-profile aneurysm clip that did not protrude through the dural defect, as well as self-tying sutures. PMID- 18424980 TI - Modifications of the transoral approach to the craniovertebral junction: anatomic study and clinical correlations. AB - OBJECTIVE: This study was designed to more precisely characterize the changes in exposure achieved by modifying the standard transoral approach by sequential mandibulotomy and mandibuloglossotomy with or without palatotomy. METHODS: A series of cadaveric dissections was performed and the operative distance and angle of exposure in both axial and sagittal planes was evaluated for each approach, with and without palatotomy. Intraoperative measurements were made in patients undergoing transoral approaches to assess the validity of the anatomic model. The use of this model was then assessed by a retrospective analysis of a group of 19 patients operated on through transoral approaches between 1991 and 2006. RESULTS: The simple transoral approach exposed the region from the lower third of the clivus to the middle of the C2 vertebral body at an operative distance of 12.9 +/- 1.0 cm from the dura. The axial and sagittal angles of exposure were 39.4 +/- 3.5 degrees and 36.8 +/- 3.5 degrees, respectively. Mandibulotomy significantly increased the sagittal exposure to 59.0 +/- 1.0 degrees (P < 0.001), exposing the area from the midclivus to the C2-C3 interspace while simultaneously increasing the axial angle of exposure to 51.9 +/- 7.4 degrees (P < 0.01) and decreasing the operative distance to the dura to 10.7 +/- 1.7 cm (P < 0.05). Mandibuloglossotomy augmented sagittal exposure to 85.3 +/- 0.3 degrees (P < 0.001), revealing the region between the upper one-third of the clivus and the C4-C5 interspace (P < 0.001) while decreasing the operative distance to the dura to 8.7 +/- 0.3 cm (P < 0.05). Palatotomy significantly increased the rostral exposure achieved by each approach by 8.5 to 12.3 degrees (P < 0.01) without altering caudal or axial exposure or the operative distance. CONCLUSION: The cadaveric data correlated well with intraoperative measurements and the need for modifications of the transoral approach in 15 of the 16 adult patients (93.8%). Pediatric patients, patients with limited mouth opening, elevated craniovertebral junctions, and particularly deep lesions required more extensive exposure. This analysis may be useful for determining the optimal approach for patients undergoing transoral surgery. PMID- 18424981 TI - Cerebrospinal fluid-related complications with autologous duraplasty and arachnoid sparing in type I Chiari malformation. AB - OBJECTIVE: Although there is a current consensus that Type I Chiari malformations (CM-I) should be treated only in the setting of symptomatic disease, significant controversy surrounds the most appropriate surgical procedure. Recent enthusiasm for osseous decompression without duraplasty is supported by the purportedly lower morbidity of this approach. Precise rates of morbidity with duraplasty, however, have not been reported. This study is intended to assess the cerebrospinal fluid-related morbidity associated with a patient population treated uniformly with autologous duraplasty for symptomatic CM-I. METHODS: A review of one surgeon's practice (MMS) from 1997 to 2007 identified patients treated for symptomatic CM-I with osseous decompression and autologous duraplasty. A retrospective chart review was then performed for these patients with an emphasis on cerebrospinal fluid-related complications. RESULTS: Forty patients were treated for CM-I with decompression and autologous duraplasty. Twenty-four patients presented with a preoperative syrinx. The mean age was 13.3 years, and the median age was 12.9 years (range, 3.3-45.8 yr). The mean follow-up period was 11.4 months (range, 1-101 mo). There was no mortality associated with the procedure. Clinical response was observed in 91.8% of patients, with 70.2% experiencing complete symptomatic resolution and 21.6% experiencing partial improvement. Two patients (5.4%) had persistent symptomatic syringomyelia requiring syringosubarachnoid shunting. There was an overall morbidity rate of 2.5% due to one pseudomeningocele treated with a single percutaneous tap. There were no incidences of cerebrospinal fluid leak, meningitis, or postoperative hydrocephalus. CONCLUSION: The cerebrospinal fluid-related morbidity associated with autologous duraplasty for CM-I in a uniformly treated population is negligible. These results challenge the current rationale for a less aggressive surgical approach to CM-I. PMID- 18424982 TI - Correction of late traumatic thoracic and thoracolumbar kyphotic spinal deformities using posteriorly placed intervertebral distraction cages. AB - OBJECTIVE: To assess the safety and stability of thoracic or thoracolumbar deformity correction from a solely posterior approach with placement of modular anterior cages and posterior segmental fixation in one operation. METHODS: Twenty eight patients who failed brace trial for 6 months or longer were included in the series. All patients had progressive neurological deficit and/or deformity progression at time of operation. All patients underwent a single operation in the prone position. Segmental fixation was accompanied by anterior column reconstruction using modular cages avoiding nerve root sacrifice. Stackable cages were used for high thoracic deformity. Deformity, Cobb angle, visual analog pain score, and x-ray evaluation of fusion ensued for mean follow-up period of 31 months. RESULTS: Patients achieved a mean sagittal deformity correction of 13.3 degrees +/- 7.4 standard deviation. Improved or maintained American Spinal Injury Association scores were noted in all patients. The mean time of operation was 334 minutes +/- 85 standard deviation, or 6 to 7 hours. At a mean follow-up of 31 months (range, 12-36 mo), the following complications were noted: subsidence greater than 2.5 mm (n = 3), cage migration requiring revision (n = 1), brachial plexopathy from malpositioning (n = 1), and intraoperative cerebrospinal fluid leak managed via lumbar drain (n = 2). Plain and dynamic radiographic evidence of maintained deformity correction was noted in 27 patients. CONCLUSION: Delayed kyphotic deformity correction of the thoracolumbar spine is achieved via a posterior-only approach. At a mean follow-up period of 31 months, sagittal angles remained acceptable. Improved fusion criteria and patient numbers will be required to determine fusion and loss of correction rates over time. PMID- 18424983 TI - The translaminar approach to lumbar disc herniations impinging the exiting root. AB - OBJECTIVE: We undertook a prospective, non-randomized study on the translaminar approach for the treatment of cephalad extruded disc fragments impinging the exiting root. METHODS: Between May 2000 and July 2004, 104 patients (59 men) presenting with upper lumbar root compression in 74% of the cases -underwent a translaminar approach. The mean age was 57 years (range, 27-80 yr). The lamina was approached either through the conventional subperiosteal route or via a muscle splitting access. Mostly intraforaminal disc fragments were removed through a translaminar hole 10 mm in diameter, and the disc space was cleared in cases of evident perforation of the annulus. Follow-up examinations were performed by an independent observer at 1 and 6 weeks; 3, 6, and 12 months; and once yearly thereafter (mean follow-up period, 32 mo). RESULTS: Extruded (61%) or subligamentous (39%) disc fragments were found intra-operatively. Laminae L4 (44%) and L5 (26%) were mostly involved. In eight cases, the translaminar hole was enlarged to a conventional laminotomy. In 13 patients, the disc space was cleared. The outcomes according to the Macnab criteria were excellent (67%), good (27%), fair (5%), and poor (1%). The incidence of recurrent disc herniations was 7%. Functional radiography performed in the first 20 patients 6 months after surgery and an additional 12 patients complaining of postsurgical back pain excluded any instability. CONCLUSION: The translaminar approach is recommended in disc herniations encroaching the exiting root, as an alternative to the conventional interlaminar route. PMID- 18424984 TI - The infraclavicular approach to the brachial plexus. AB - The infraclavicular approach to the brachial plexus is commonly indicated in patients with traumatic injuries and tumors of the brachial plexus elements. We describe the anatomy and operative technique of this approach. PMID- 18424985 TI - Surgical approach to ulnar nerve compression at the elbow caused by the epitrochleoanconeus muscle and a prominent medial head of the triceps. AB - OBJECTIVE: We sought to describe the operative technique in ulnar nerve compression caused by the epitrochleoanconeus muscle and a prominent medial head of the triceps. These anatomic features make the approach to the ulnar nerve at the elbow peculiar and may create technical difficulties during surgical treatment of this area. METHODS: We reviewed patients who underwent surgery for cubital tunnel syndrome between November 1997 and December 2004. The presence of the epitrochleoanconeus muscle with prominent medial head of the triceps occurred in 3.2% of patients. A detailed and illustrated description of the surgical anatomy and the peculiarities of the surgical approach are provided. RESULTS: Epitrochleoanconeus muscle and the prominent portion of the medial head of the triceps were sectioned and removed, and simple decompression of the ulnar nerve was performed. This treatment achieved complete recovery in all of the patients affected by moderate-grade syndrome (Dellon Grade 2 syndrome) who had not shown severe-grade syndrome preoperatively. CONCLUSION: The simple decompression of the ulnar nerve with myotomy or removal of epitrochleoanconeus muscle and the prominent portion of the medial head of the triceps achieved good postoperative results. Experiences from the literature and alternative surgical options are reported. PMID- 18424986 TI - Clinical and electrophysiological comparison of different methods of soft tissue coverage of the median nerve in recurrent carpal tunnel syndrome. AB - OBJECTIVE: To evaluate the clinical and electrophysiological results of 26 patients treated with either a hypothenar fat flap or a synovial flap to prevent recurrent scar compression of the median nerve after previously failed carpal tunnel decompression. METHODS: A total of 26 patients underwent flap coverage as a result of a nerve tethering attributable to a position within scar; 15 were covered by a synovial flap and 11 by a hypothenar fat flap. Only patients in whom the median nerve was significantly enveloped in scar tissue were included. All candidates underwent a thorough clinical examination and nerve conduction test. The pre- and postoperative nerve conduction tests and the results of the two groups were statistically compared. RESULTS: The reduction rates of brachial nocturnal pain and pillar pain were 25 and 25%, respectively, in the synovial flap group and 64 and 37%, respectively, in the hypothenar fat flap group. The reduction rates of a positive Tinel's sign (25%) and a positive Phalen's test (13%) were lower in the synovial flap group compared with hypothenar fat flap coverage (55% Tinel's sign, 46% Phalen's test). Thenar atrophy and paresthesia were reduced in 44 and 62%, respectively, in the synovial flap group and in 46 and 64%, respectively, in the hypothenar fat flap group. The overall patient satisfaction (73%) and the Disabilities of the Arm, Shoulder and Hand score (31 points) appeared superior in the hypothenar fat flap group compared with the synovial flap group (56%; 37 points). Nerve conduction tests demonstrated a significant improvement when comparing the pre- and postoperative measurements in both groups. Distal motor latency decreased in the hypothenar fat flap group from 6.81 ms to 4.92 msec (P = 0.01; mean value) and in the synovial flap group from 6.04 ms to 4.43 msec (P < 0.001; mean value). CONCLUSION: Coverage by an ulnar based hypothenar fat flap appeared to produce superior clinical results compared with coverage with synovial tissue from adjacent flexor tendons, although conclusive statistical evaluation of clinical outcomes was not possible. Further studies to confirm this are warranted. PMID- 18424987 TI - Fiducial versus nonfiducial neuronavigation registration assessment and considerations of accuracy. AB - OBJECTIVE: For frameless stereotaxy, users can choose between anatomic landmarks (ALs) or surface fiducial markers (FMs) for their match points during registration to define an alignment of the head in the physical and radiographic image space. In this study, we sought to determine the concordance among a point merged FM registration, a point-merged AL registration, and a combined point merged anatomic/surface-merged (SM) registration, i.e., to determine the accuracy of registration techniques with and without FMs by examining the extent of agreement between the system-generated predicted value and physical measured values. METHODS: We examined 30 volunteers treated with gamma knife surgery. The frameless stereotactic image-guidance system called the StealthStation (Medtronic Surgical Navigation Technologies, Louisville, CO) was used. Nine FMs were placed on the patient's head and four were placed on a Leksell frame rod-box, which acted as a rigid set to determine the difference in error. For each registration form, we recorded the generated measurement (GM) and the physical measurement (PM) to each of the four checkpoint FMs. Bland and Altman plot difference analyses were used to compare measurement techniques. Correlations and descriptive analyses were completed. RESULTS: The mean of values for GMs were 1.14 mm for FM, 2.3 mm for AL, and 0.96 mm for SM registrations. The mean errors of the checkpoints were 3.49 mm for FM, 3.96 mm for AL, and 3.33 mm for SM registrations. The correlation between GMs and PMs indicated a linear relationship for all three methods. AL registration demonstrated the greatest mean difference, followed by FM registration; SM registration had the smallest difference between GMs and PMs. Differences in the anatomic registration methods, including SM registration, compared with FM registration were within a mean +/- 1.96 (standard deviation) according to the Bland and Altman analysis. CONCLUSION: For our sample of 30 patients, all three registration methods provided comparable distances to the target tissue for surgical procedures. Users may safely choose anatomic registration as a less costly and more time-efficient registration method for frameless stereotaxy. PMID- 18424988 TI - Compensation of geometric distortion effects on intraoperative magnetic resonance imaging for enhanced visualization in image-guided neurosurgery. AB - OBJECTIVE: Preoperative magnetic resonance imaging (MRI), functional MRI, diffusion tensor MRI, magnetic resonance spectroscopy, and positron-emission tomographic scans may be aligned to intraoperative MRI to enhance visualization and navigation during image-guided neurosurgery. However, several effects (both machine- and patient-induced distortions) lead to significant geometric distortion of intraoperative MRI. Therefore, a precise alignment of these image modalities requires correction of the geometric distortion. We propose and evaluate a novel method to compensate for the geometric distortion of intraoperative 0.5-T MRI in image-guided neurosurgery. METHODS: In this initial pilot study, 11 neurosurgical procedures were prospectively enrolled. The scheme used to correct the geometric distortion is based on a nonrigid registration algorithm introduced by our group. This registration scheme uses image features to establish correspondence between images. It estimates a smooth geometric distortion compensation field by regularizing the displacements estimated at the correspondences. A patient-specific linear elastic material model is used to achieve the regularization. The geometry of intraoperative images (0.5 T) is changed so that the images match the preoperative MRI scans (3 T). RESULTS: We compared the alignment between preoperative and intraoperative imaging using 1) only rigid registration without correction of the geometric distortion, and 2) rigid registration and compensation for the geometric distortion. We evaluated the success of the geometric distortion correction algorithm by measuring the Hausdorff distance between boundaries in the 3-T and 0.5-T MRIs after rigid registration alone and with the addition of geometric distortion correction of the 0.5-T MRI. Overall, the mean magnitude of the geometric distortion measured on the intraoperative images is 10.3 mm with a minimum of 2.91 mm and a maximum of 21.5 mm. The measured accuracy of the geometric distortion compensation algorithm is 1.93 mm. There is a statistically significant difference between the accuracy of the alignment of preoperative and intraoperative images, both with and without the correction of geometric distortion (P < 0.001). CONCLUSION: The major contributions of this study are 1) identification of geometric distortion of intraoperative images relative to preoperative images, 2) measurement of the geometric distortion, 3) application of nonrigid registration to compensate for geometric distortion during neurosurgery, 4) measurement of residual distortion after geometric distortion correction, and 5) phantom study to quantify geometric distortion. PMID- 18424989 TI - Location of active contacts in patients with primary dystonia treated with globus pallidus deep brain stimulation. AB - OBJECTIVE: Deep brain stimulation of the globus pallidus internus has been used for the treatment of various forms of dystonia, but the factors influencing postoperative outcomes remain unknown. We compared the location of the contacts being used for stimulation (active contacts) in patients with cervical dystonia, generalized dystonia, and Parkinson's disease and correlated the results with clinical outcome. METHODS: Postoperative magnetic resonance scans of 13 patients with cervical dystonia, six patients with generalized dystonia, and five patients with Parkinson's disease who underwent globus pallidus internus deep brain stimulation were analyzed. We assessed the location of the active contacts relative to the midcommisural point and in relation to the anteroposterior and mediolateral boundaries of the pallidum. Postoperative outcome was measured with the Toronto Western Spasmodic Torticollis Rating Scale (for cervical dystonia) and the Burke-Fahn-Marsden Dystonia Rating Scale (for generalized dystonia) during the last follow-up. RESULTS: We found that the location of the active contacts relative to the midcommisural point and the internal boundaries of the pallidum was similar across the groups. In our series, the contacts used for stimulation were clustered in the posterolateral region of the pallidum. Within that region, we found no correlation between the location of the contacts and postoperative outcome. CONCLUSION: The location of the active contacts used for globus pallidus internus deep brain stimulation was similar in patients with cervical dystonia, generalized dystonia, and Parkinson's disease. PMID- 18424990 TI - Percutaneous computed tomography-guided radiofrequency ablation of upper spinal cord pain pathways for cancer-related pain. AB - OBJECTIVE: The author presents data to support the continued need for ablative procedures, particularly cordotomy, in the management of cancer-related pain. METHODS: Fifty-one patients with cancer-related body or face pain were treated with computed tomography-guided radiofrequency ablation of the spinothalamic tract or trigeminal tract nucleus in the upper cervical region of the spinal cord. Forty-one patients underwent a unilateral cervical cordotomy, and 10 patients underwent a trigeminal tractotomy-nucleotomy. Three methods to assess patient pain were used: degree of pain relief, Visual Analog Scale, and total sleeping hours. The Karnofsky scale was used to measure the patient's level of function pre- and postprocedure. RESULTS: After surgical intervention, patients reported initial and 6-months follow-up pain relief as 98 and 80%, respectively. CONCLUSION: Computed tomography-guided ablation of the upper cervical spinal cord is a safe and effective procedure to treat cancer pain involving the body or face. There remains a need for ablative procedures, in particular cordotomy, in the management of cancer-related pain. PMID- 18424991 TI - Spinal and nucleus caudalis dorsal root entry zone operations for chronic pain. AB - OBJECTIVE: Dorsal root entry zone (DREZ) operations came into medical practice after the demonstration of increased electrical activity in the dorsal horn of the spinal cord and brainstem in patients with deafferentation of the central nervous system after injury to these areas. The aim of the study was to describe the technique and the effectiveness of spinal DREZ and nucleus caudalis (NC) DREZ operations, which may be the treatments of choice in unique chronic pain conditions that do not respond to medical therapy or any other surgical methods. METHODS: Fifty-five patients (44 spinal, 11 NC DREZ) underwent 59 (48 spinal, 11 NC DREZ) operations. There were 44 men and 11 women with a mean age of 46.4 years (range, 24-74 yr). The mean follow-up period was 72 months (range, 6 mo-20 yr). Follow-up assessments were performed with clinical examination on the first day and in the sixth and twelfth months postoperatively. Patients' pain scores and Karnofsky Performance Scale scores were also evaluated pre- and postoperatively. RESULTS: The initial success rates for spinal and NC DREZotomy procedures were 77 and 72.5%, respectively. In the spinal DREZotomy group, mortality occurred in one patient (2.2%). There were two cases of transient muscle weakness (4.4%) and two of cerebrospinal fluid fistulae (4.4%). In the NC DREZotomy group, mortality occurred in one patient (9%). There were two cases of transient ataxia (18%) and two of transient hemiparesis (18%). CONCLUSION: Spinal and trigeminal NC DREZ operations are effective in the treatment of intractable pain syndromes, especially in traumatic brachial plexus avulsions, segmental pain after spinal cord injury, postherpetic neuralgia, topographically limited cancer pain, and atypical facial pain. PMID- 18424992 TI - Use of an integrated platform system in the placement of deep brain stimulators. AB - The placement of deep brain stimulator leads requires a great deal of technology and equipment. We describe our 25-month experience with an integrated platform system, the StimPilot (Medtronic Inc., Minneapolis, MN), for the placement of deep brain stimulator leads. The platform consists of a neuronavigation station, microdrive control, and microelectrode recording display and control. This platform is run from a laptop-sized portable control unit. The unit was used in 147 patients for the placement of 262 leads. Leads were placed into the subthalamic nucleus, ventral intermediate nucleus, globus pallidus interna, and anterior thalamic nucleus. One patient required replacement of one lead during this time frame, with successful reimplantation. No system failures occurred. PMID- 18424993 TI - Methods of scalp revision for deep brain stimulator hardware: case report. AB - OBJECTIVE: The use of deep brain stimulation (DBS) to treat a variety of disorders has expanded and will result in an increasingly larger number of patients and implanted electrodes. Hardware failure can result from malfunction, lead migration, fracture, and infection. Scalp erosion with exposure of underlying hardware can lead to potential infectious complications and is, in itself, a strong indication for explantation of the neurostimulation system. The patient's relief of symptoms after DBS will be limited by hardware-related complications and thus, strategies to revise scalp overlying hardware are important in the widespread application of DBS. CLINICAL PRESENTATION: We describe strategies to address complications related to implanted DBS neurostimulator hardware specifically designed to address breach of the integrity of the scalp over the burr hole site. The aim of these approaches is to treat scalp erosion to allow for the reimplantation of previously explanted, infected hardware, or to treat thinned scalp with threatened erosion and prevent the need to remove exposed hardware that is otherwise functioning. INTERVENTION: Two different approaches are presented: 1) a temporoparieto-occipital flap based on the superficial temporal artery with or without scalp expansion, and 2) a scalp fasciocutaneous flap with or without cranioplasty. CONCLUSION: Stimulation of various deep brain targets helps patients with a wide range of diseases. In the future, with continued refinement, hardware complications can be minimized. Until then, novel approaches need to be developed to save DBS systems and provide symptomatic relief to patients. PMID- 18424994 TI - Quantification of the frontotemporal orbitozygomatic approach using a three dimensional visualization and modeling application. AB - OBJECTIVE: We sought to simulate the frontotemporal orbitozygomatic (FTOZ) craniotomy in a three-dimensional virtual environment on patient-specific data and to quantify the exposure afforded by the FTOZ while simulating controlled amounts of brain retraction. METHODS: Four computed tomographic angiograms were reconstructed with commercially available software (Amira 4.1.1; Mercury Computer Systems, Inc., Chelmsford, MA), and virtual FTOZ craniotomies were performed bilaterally (n = 8). Brain retraction was simulated at 1 and 2 cm. Surgical freedom and projection angle were measured and compared at each stage of the FTOZ. RESULTS: At 1 cm of retraction, surgical freedom increased by 27 +/- 14% for the removal of the orbital rim and by 31 +/- 18% for FTOZ (P < 0.01) when compared with frontotemporal (FT) craniotomy. At 2 cm of retraction, surgical freedom increased by 15 +/- 5% and 26 +/- 8% for the removal of the orbital rim and FTOZ, respectively (P < 0.01). With increased retraction, surgical freedom increased by 100 +/- 26%, 81 +/- 15%, and 82 +/- 27% for the FT, removal of the orbital rim, and FTOZ craniotomies, respectively (P < 0.001). Projection angle increased by 24.2% when orbital rim removal was added to the FT craniotomy (P < 0.01). CONCLUSION: Surgical freedom increases significantly at every step of the FTOZ craniotomy. This effect is less robust when brain retraction is increased. Brain retraction alone has a greater impact on surgical freedom than bone removal alone. Projection angle is significantly increased when orbital rim removal is added to the FT craniotomy. This model overcomes two major limitations of cadaver based models: quantification of brain retraction and incorporation of patient specific anatomy. PMID- 18424995 TI - Frontozygomatic titanium cranioplasty in frontosphenotemporal ("pterional") craniotomy. AB - OBJECTIVE: One of the most common problems after frontosphenotemporal, or pterional, craniotomy is the marked depression of the frontozygomatic fossa caused by atrophy of the temporalis muscle. Although temporalis muscle reconstruction techniques have been proposed to prevent this problem, a definitive solution has not been achieved. We report the results of a titanium cranioplasty technique in a prospective series of patients who underwent frontosphenotemporal craniotomy. METHODS: Between April 2002 and June 2006, 209 consecutive patients underwent a frontosphenotemporal craniotomy for aneurysms, vascular malformations, or tumors. At the time of surgery, the patients underwent a frontozygomatic fossa cranioplasty with a titanium plate, to which the temporalis muscle was attached. In this series, 194 patients had documented follow-up periods averaging 9.5 months (range, 1 mo-4 yr; median, 7.5 mo), and the cosmetic results of the cranioplasty have been assessed. RESULTS: The cosmetic outcomes have been outstanding in all patients treated to date. Two patients had the cranioplasty removed due to either orbital pain or local infection secondary to sepsis. CONCLUSION: The frontozygomatic cranioplasty during frontosphenotemporal craniotomy prevents the characteristic depression at the frontozygomatic fossa and accomplishes an outstanding cosmetic result. PMID- 18424996 TI - Dynact soft-tissue visualization using an angiographic C-arm system: initial clinical experience in the operating room. AB - INTRODUCTION: DynaCT is a clinical application protocol to create computed tomographic (CT)-like images allowing soft-tissue visualization acquired from an angiography system. A cone beam three-dimensional CT reconstruction is produced from the acquisition of two-dimensional projection images by rotating the c-arm with x-ray source and image receptor around the patient. The purpose of this study is to evaluate the clinical efficacy of DynaCT in the operating room. METHODS: DynaCT was performed in 100 patients undergoing cerebral or spinal interventional procedures in the new-concept operating room. Specially designed AXIOM Artis BA (conventional image intensifier system; 55 patients; Siemens Medical Solutions, Erlangen, Germany) and AXIOM Artis dBA (flat-panel detector; 45 patients; Siemens Medical Solutions) biplane neuroangiographic systems (Siemens Medical Solutions, Germany) were installed in the operating room. The volumetric data set from the AXIOM Artis systems were reconstructed immediately on the three-dimensional workstation in the operating room. We compared DynaCT images with the use of multidetector computed tomography. RESULTS: DynaCT was performed successfully in all patients. High-contrast structures, such as bone, calcified lesions, and metallic materials, were visualized on DynaCT as well as in multidetector computed tomography for each group. Although contrast differentiation of soft tissue such as cerebral cortex, muscle, and hematoma on DynaCT were inferior to multidetector CT scans, the images were sufficiently used as intraoperative CT-like images. However, DynaCT images acquired from flat panel detector-based systems were found to be superior to those images acquired from image intensifier-based systems. Striking ring artifacts were exhibited and resulted in major limitations in the image intensifier group. CONCLUSION: DynaCT has the potential to be used as a powerful tool for endovascular and neurosurgical procedures and will open new possibilities for neurosurgical management. PMID- 18424997 TI - Novel equine collagen-only dural substitute. AB - OBJECTIVE: A watertight and meticulous dural closure is an essential step after intradural neurosurgical procedures. When such a task cannot be performed, dural replacement materials and other adjunctive measures can provide an effective barrier between the subarachnoid compartment and the extradural space. METHODS: We present our experience with a novel collagen-derived dural substitute in a series of 114 patients undergoing a variety of neurosurgical procedures. The patients were clinically or neuroradiologically observed, for immediate and delayed local or systemic complications related to the implant. In three patients who underwent reoperation after decompressive duraplasty and craniectomy for bone flap repositioning, we performed biopsy of the dural implant for histopathological studies. RESULTS: None of the patients experienced local or systemic complications or toxicity related to the dural patch. None of the patients experienced a postoperative cerebrospinal fluid fistula, except one patient who underwent an endoscopic endonasal transsphenoidal marsupialization of a large intrasuprasellar arachnoid cyst; the fistula required reoperation for cerebrospinal fluid fistula repair and intravenous antibiotic therapy for bacterial meningitis. Postoperative magnetic resonance imaging scans showed signs of severe inflammatory response in only one patient who did not present any postoperative clinical symptom or neurological deficits. Three patients underwent reoperation for bone flap repositioning after decompressive craniectomy; in all patients, the dural substitute appeared to have promoted satisfactory dural regeneration, as confirmed by the histological studies. Furthermore, in such patients, no or minimal adherence with the other tissues and the brain cortex was observed. CONCLUSION: This study demonstrates that the new collagen-only biomatrix is a safe and effective dural substitute for routine neurosurgical procedures. The absence of local and systemic toxicity or complications and the scarce promotion of adherences and inflammation make this material appealing for its use as a dural substitute, even in cases in which the necessity of reoperation is foreseen. PMID- 18424999 TI - Radiation-induced meningioma concealed by shunt valve artifact: case report. AB - OBJECTIVE: Ventricular shunting is a common neurosurgical procedure. Metal containing shunt components can cause artifacts on magnetic resonance imaging (MRI), obscuring visualization of the surrounding tissue. We present the case of a radiation-induced meningioma growing around a ventricular catheter that was not noted at an early stage as a result of artifact from an overlying shunt. CLINICAL PRESENTATION: We present the case of a 27-year-old survivor of a childhood medulloblastoma who required ventriculoperitoneal shunting as a consequence of her treatment. As a result of radiation-induced meningiomas that developed in adolescence, the patient was screened biennially via contrasted MRI. Her most recent imaging revealed a new meningioma at the edge of the MRI artifact associated with the shunt valve. A contrasted computed tomographic scan demonstrated a large meningioma with mass effect on the surrounding brain. INTERVENTION: Complete surgical resection of the meningioma was obtained. The ventricular catheter was preserved and the shunt valve replaced with a newer system that is reported to generate less magnetic artifact. CONCLUSION: Artifact from shunt valves or other implanted metallic devices obscures the surrounding tissues on MRI. Patients with significant artifact who are at higher risk for development of intracranial pathology may benefit from periodic imaging through alternate modalities that are not susceptible to magnetic artifact. PMID- 18425000 TI - Myelolipomatous change in an interhemispheric lipoma associated with corpus callosum agenesis: case report. AB - OBJECTIVE: Interhemisperic lipomas are almost always associated with hypogenesis or agenesis of the corpus callosum. These lesions are stable. CLINICAL PRESENTATION: We report a case of an interhemispheric lipoma in a 72-year-old man who had undergone myelolipomatous differentiation. INTERVENTION: Gross total excision of the lesion was performed. CONCLUSION: Although extremely uncommon, the possibility of myelomatous change should be considered in the differential diagnosis of an intracranial lipoma if it shows some areas of mixed density. PMID- 18425001 TI - Persistent trigeminal artery terminating in the posterior inferior cerebellar artery: case report. AB - OBJECTIVE: Persistent trigeminal arteries are rare and represent a remnant of the fetal carotid-basilar circulation. They typically extend from the internal carotid artery to the basilar artery. An unusual case of a patient with a trigeminal artery originating from the internal carotid artery and terminating as the dominant hemispheric branch of the posterior inferior cerebellar artery is presented. CLINICAL PRESENTATION: A 66-year-old woman presented to the emergency department with pulsatile tinnitus, increasing left eye pain, proptosis, conjunctival injection, diplopia, and decreased visual acuity. Conventional contrast-enhanced computed tomographic and magnetic resonance imaging demonstrated findings consistent with a left carotid-cavernous fistula. The patient underwent an emergency diagnostic cerebral angiogram. Besides an indirect carotid-cavernous fistula on the left side, a right-sided persistent trigeminal artery terminating as the dominant hemispheric trunk of the posteroinferior cerebellar artery was incidentally noted. The vermian branch of the right posteroinferior cerebellar artery arose from the ipsilateral vertebral artery, whereas duplicate superior cerebellar arteries supplied the left posteroinferior cerebellar artery region. INTERVENTION: The patient was treated for the indirect carotid-cavernous fistula with detachable platinum coils and N-butyl cyanoacrylate, resulting in the resolution of her symptoms. CONCLUSION: We report a case of a persistent trigeminal artery supplying only the cerebellar hemisphere. The clinical significance of this anomaly relates to its role in endovascular therapeutic and surgical complications and the paradoxical lesions in the cerebellum that occur as a result of carotid disease. We also discuss the Saltzman classification of persistent trigeminal arteries and their variants. PMID- 18425002 TI - Survival with good outcome after traversing brainstem injury: case report. AB - OBJECTIVE: A penetrating injury to the brainstem is usually a fatal injury. The number of cases in which the brainstem is traversed during the injury and the patient survives is exceedingly small. CLINICAL PRESENTATION: We report a case of an automobile collision in which blunt injury to the face of a 22-year-old man resulted in a fragment of the clivus being impacted posteriorly traversing through the pons in a left-sided through-and-through manner. The striking radiographic images demonstrate a potentially devastating brainstem injury. Surprisingly, the patient was able to follow commands and move his left side on arrival at the hospital. INTERVENTION: The patient required a temporary tracheostomy and underwent surgical repair of his facial fractures. His hospital course was complicated by meningitis, and he required a long stay on the rehabilitation service. The patient made a remarkable recovery and became able to converse and walk with assistance. He has been able to live independently. CONCLUSION: The striking radiographic images of this rare case illustrate an unusual pathology with an even more unusual outcome. PMID- 18425004 TI - Proteomics: present and future implications in neuro-oncology. AB - PROTEOMICS, IN ITS broadest mandate, is the study of proteins and their functions. As the "workhorses" of the genome, proteins govern normal cellular structure and function. Protein function is not just a reflection of its expression level; it is also the cumulative result of many post-transcriptional (splicing) and post-translational events that together determine cellular localization, interactions, and longevity. The composition and variability of the proteome is vastly more complex than the corresponding genome. It is this proteome variation that helps define an organism and the unique characteristics that separate one individual from another. Aberrations in protein function, which alter normal cellular structure and function, are the ultimate basis of disease, including cancer. Therefore, an understanding of protein networks through a systems biology approach of proteomics is necessary to understand normal and abnormal cellular function, with the goal of performing rational therapeutic interventions. In this review, we focus on two emerging proteomic technologies: mass spectrometry and bioluminescence resonance energy transfer. In addition to reviewing the principles and potential utilization of these two techniques, we highlight their application in neuro-oncology research. PMID- 18425005 TI - Tuberculum sellae meningiomas: high route or low route? A series of 51 consecutive cases. AB - OBJECTIVE: Tuberculum sellae meningiomas represent 5 to 10% of all intracranial meningiomas. Such lesions are classically removed through a variety of well standardized transcranial approaches. The extended endonasal transsphenoidal route, under either microscopic or endoscopic visualization, has only recently been proposed as a viable surgical technique for the management of such tumors. MATERIAL AND METHODS: A total of 51 consecutive patients with tuberculum sellae meningiomas were treated at our institution during a 21-year period. Forty-four patients had transcranial surgery, and the last seven were treated via the extended endoscopic transsphenoidal approach. We also compared our data with those reported in the pertinent literature related to the surgical, ophthalmological, and endocrinological outcome. RESULTS: The significant difference among the transcranial and transsphenoidal series, both in our experience and in the reviewed literature, did not allow us to draw statistically significant results but rather a reporting of the outcomes. In the transcranial group, 86.4% had a gross total removal of the lesion, whereas the percentage was 83.3% in the transsphenoidal group. Concerning the visual outcome, we experienced postoperative improvement in 61.4% of the transcranial patients and a worsening of 13.6%, whereas improvement was reported in 71.4% of the patients in the transsphenoidal group; in the last group, we did not observe any postoperative worsening. The main drawback of the transsphenoidal approach still remains the difficulty in reconstructing the cranial base dural and bone defects, which expose patients to a greater risk of postoperative cerebrospinal fluid leakage (28.6% in our series) and related complications. CONCLUSION: When treating a patient with a diagnosis of tuberculum sellae meningioma, a neurosurgeon should know that, aside from the classical transcranial approach, the possibility of an extended transsphenoidal approach exists. Although it is still not a standardized procedure, in carefully selected cases (i.e., small midline lesions, without major vessel encasement, or parasellar extension) and in experienced hands, it could be considered a viable alternative, especially in overcoming the reconstruction-related problems. PMID- 18425007 TI - Localization of cerebrospinal fluid leaks by gadolinium-enhanced magnetic resonance cisternography: a 5-year single-center experience. AB - OBJECTIVE: Intrathecal gadolinium (Gd)-enhanced magnetic resonance (MR) cisternography is a newly introduced imaging method. Two main objectives of this study were to investigate the sensitivity of Gd-enhanced MR cisternography for presurgical localization of cerebrospinal fluid (CSF) leaks in patients with CSF rhinorrhea and to study the potential long-term adverse effects of intrathecal Gd application. METHODS: Fifty-one patients (19 women; mean age, 36.2 yr) with CSF rhinorrhea were included in the study. A total of 0.5 ml of Gd was injected into the lumbar subarachnoid space. T1-weighted MR cisternographic images were obtained to detect CSF leakage. The patient's neurological states and vital signs were recorded for the first 24 hours after the procedure. Neurological evaluations were repeated 1, 3, and 12 months after the procedure. The patients were followed for at least 3 years with annual neurological examinations. RESULTS: Gd-enhanced MR cisternography demonstrated CSF leaks in 43 of the 51 patients. The sensitivity of Gd-enhanced MR cisternography for localization of CSF leaks was 84%. Forty-four patients underwent surgery to repair dural tears. Surgical findings confirmed the results of Gd-enhanced cisternography in 43 of the 44 patients who underwent surgery (98%). Eight patients with negative Gd enhanced MR cisternography had no active rhinorrhea at the time of procedure, and seven of them did not need surgery. None of the patients developed an acute adverse reaction that could be attributed to the procedure. None of the patients developed any neurological symptoms or signs caused by intrathecal Gd injection during a mean follow-up period of 4.12 years. CONCLUSION: Gd-enhanced MR cisternography is a sensitive and safe imaging method for detection of CSF leaks in patients with rhinorrhea. PMID- 18425006 TI - Extent of resection and survival in glioblastoma multiforme: identification of and adjustment for bias. AB - OBJECTIVE: The influence of the degree of resection on survival in patients with glioblastoma multiforme is still under discussion. The highly controlled 5 aminolevulinic acid study provided a unique platform for addressing this question as a result of the high frequency of "complete" resections, as revealed by postoperative magnetic resonance imaging scans achieved by fluorescence-guided resection and homogeneous patient characteristics. METHODS: Two hundred forty three patients with glioblastoma multiforme per protocol from the 5 aminolevulinic acid study were analyzed. Patients with complete and incomplete resections as revealed by early magnetic resonance imaging scans were compared. Prognostic factors that might cause bias regarding resection and influence survival (e.g., tumor size, edema, midline shift, location, age, Karnofsky Performance Scale score, National Institutes of Health Stroke Scale score) were used for analysis of overall survival. Time to reintervention (chemotherapy, reoperation) was analyzed further to exclude bias regarding second-line therapies. RESULTS: Treatment bias was identified in patients with complete (n = 122) compared with incomplete resection (n = 121), i.e., younger age and less frequent eloquent tumor location. Other factors, foremost preoperative tumor size, were identical. Patients without residual tumor survived longer (16.7 versus 11.8 mo, P < 0.0001). In multivariate analysis, only residual tumor, age, and Karnofsky Performance Scale score were significantly prognostic. To account for distribution bias, patients were stratified for age (>60 or or=4) as other ruptured aneurysms (74 versus 69%), but their mortality rate was lower (13 versus 24%). Factors predicting unfavorable outcome for ruptured DACA aneurysms were advanced age, Hunt and Hess grade greater than or equal to III, rebleeding before treatment, intracerebral hematoma, intraventricular hemorrhage, and severe preoperative hydrocephalus. CONCLUSION: Despite their specific features, with modern treatment methods, ruptured DACA aneurysms have the same favorable outcome and lower mortality at 1 year as ruptured aneurysms in general. PMID- 18425009 TI - Ventriculoperitoneal shunting after aneurysmal subarachnoid hemorrhage: analysis of the indications, complications, and outcome with a focus on patients with borderline ventriculomegaly. AB - OBJECTIVE: The goals of this study were to investigate the risk factors, indications, complications, and outcome for patients with ventriculoperitoneal shunts (VPSs) after subarachnoid hemorrhage and to define a subgroup eligible for future prospective studies designed to clarify indications for placement of a VPS. METHODS: Clinical characteristics of 236 prospectively evaluated patients with subarachnoid hemorrhage and 6 months of follow-up were analyzed. Hydrocephalus was estimated by the relative bicaudate index (RBCI) measured on computed tomographic scans at the time of shunting. Patients were divided into three groups by ventricle size: Group 1 included 121 patients with small ventricles (RBCI <1.0), Group 2 included 88 patients with borderline ventricle size (RBCI 1.0-1.4), and Group 3 included 27 patients with markedly enlarged ventricles (RBCI >1.4). RESULTS: Initially, 86 patients (36%) underwent ventriculoperitoneal shunting: 19 in Group 1 (16%), 43 in Group 2 (49%), and 24 in Group 3 (90%). Indications for placement of a VPS, risk factors, and outcome differed markedly by group. Four patients (3% of those not initially shunted) developed delayed hydrocephalus requiring a VPS, including one in Group 2 (2%). The 6-month shunt complication rate was 13%. Evaluation of patients in Group 2 indicated that functional status was an important factor in selecting candidates for shunting, and that patients receiving shunts and shunt-free patients demonstrated improvement in functional status during follow-up. CONCLUSION: Although we currently use a proactive shunting paradigm for posthemorrhagic hydrocephalus, this report demonstrates that a conservative approach to patients with borderline ventricle size (i.e., RBCI of 1.0-1.4) and normal intracranial pressure should be evaluated in a prospective randomized trial. PMID- 18425010 TI - Risk factors for hemorrhagic presentation in patients with dural arteriovenous fistulae. AB - OBJECTIVE: Intracranial dural arteriovenous fistulae (DAVFs) can present as disabling intracranial hemorrhage. The aim of this study was to investigate the independent effects of specific demographic and clinical variables on hemorrhagic presentation in patients with DAVFs. METHODS: All patients with DAVFs evaluated at the University of California at San Francisco from July 1988 through June 2004 were identified. Clinical and radiographic characteristics were recorded using a detailed abstraction form. RESULTS: A total of 402 patients with DAVFs were identified, 73 (18%) of whom presented with intracranial hemorrhage. Men were twice as likely to present with hemorrhage (men 70% versus women 30%, P < 0.001). Cortical venous drainage (85 versus 22%; P < 0.001), retrograde venous drainage (59 versus 36%; P < 0.001), and sinus occlusion (33 versus 18%; P = 0.004) were also more common in patients with DAVF with hemorrhagic presentation. In multivariate logistic regression analysis, cortical venous drainage (odds ratio [OR], 10.5; P < 0.001), focal neurological deficits (OR, 4.7; P < 0.001), DAVFs in the posterior fossa (OR, 4.0; P = 0.005), male sex (OR, 3.4, P = 0.001), and age older than 50 years were found to be independently associated with hemorrhagic presentation. CONCLUSION: Although DAVFs are less frequent in men than in women, they are more likely to present with hemorrhage. In addition to cortical venous drainage, a well-known risk factor for intracranial hemorrhage, posterior fossa location, older age at presentation, and focal neurological deficits were independently associated with hemorrhagic presentation in patients with DAVFs. PMID- 18425011 TI - Cyberknife radiosurgery for trigeminal neuralgia treatment: a preliminary multicenter experience. AB - OBJECTIVE: Radiosurgery has gained acceptance as a treatment option for trigeminal neuralgia. We report our preliminary multicenter experience treating trigeminal neuralgia with the CyberKnife (Accuray, Inc., Sunnyvale, CA). METHODS: A total of 95 patients were treated for idiopathic trigeminal neuralgia between May 2002 and October 2005. Radiosurgical dose and volume parameters were retrospectively analyzed in relation to pain response, complications, and recurrence of symptoms. Optimal treatment parameters were identified for patients who had excellent and sustained pain relief with no complications, including severe or moderate hypesthesia. RESULTS: Excellent pain relief was initially experienced by 64 out of 95 patients (67%). The median time to pain relief was 14 days (range, 0.3-180 d). Posttreatment numbness occurred in 45 (47%) of the patients treated. Using higher radiation doses and treating longer segments of the nerve led to both better pain relief and a higher incidence of hypesthesia. The presence of posttreatment numbness was predictive of better pain relief. The overall rate of complications was 18%. At the mean follow-up time of 2 years, 47 of the 95 patients (50%) had sustained pain relief, all of whom were completely off pain medications. CONCLUSION: The results of this study suggest the following optimal radiosurgical treatment parameters for treatment of idiopathic trigeminal neuralgia: a median maximal dose of 78 Gy (range, 70-85.4 Gy) and a median length of the nerve treated of 6 mm (range, 5-12 mm). PMID- 18425012 TI - Results of end-to-side nerve coaptation in severe obstetric brachial plexus lesions. AB - OBJECTIVE: Options for nerve repair are limited in brachial plexus lesions with multiple root avulsions because an insufficient number of proximal nerve stumps are available to serve as lead-out for nerve grafts. End-to-side nerve repair might be an alternative surgical technique for repair of such severe lesions. In this technique, an epineurial window is created in a healthy nerve, and the distal stump of the injured nerve is coapted to this site. Inconsistent results of end-to-side nerve repairs in traumatic nerve lesions in adults have been reported in small series. This article evaluates the results of end-to-side nerve repair in obstetric brachial plexus lesions and reviews the literature. METHODS: A retrospective analysis was performed of 20 end-to-side repairs in 12 infants. Evaluation of functional recovery of the target muscle was performed after at least 2 years of follow up (mean, 33 mo). RESULTS: Five repairs failed (25%). Seven times (35%) good function (Medical Research Council at least 3) of the target muscle occurred in addition to eight partial recoveries (40%). In the majority of patients, however, the observed recovery cannot be exclusively attributed to the end-to-side repair. The reinnervation may be based on axonal outgrowth through grafted or neurolyzed adjacent nerves. It seems likely that recovery was solely based on the end-to-side repair in only two patients. No deficits occurred in donor nerve function. CONCLUSION: This study does not convincingly show that the end-to-side nerve repair in infants with an obstetric brachial plexus lesion is effective. Its use cannot be recommended as standard therapy. PMID- 18425013 TI - Surgical treatment of traumatic peroneal nerve lesions. AB - OBJECTIVE: In this study, we compare different surgical procedures regarding the functional outcome of traumatic peroneal nerve lesions. METHODS: In a retrospective study, 48 patients with traumatic lesions (17 iatrogenic) of the peroneal nerve were evaluated. Twenty-two patients presented with lesions in continuity displaying regenerative potential by nerve action potential recording. In these cases, surgery was restricted to either external (12/) or interfascicular neurolysis (10/). Twenty-two cases had no regenerative potential (10/) or showed discontinuity (12/) and thus were reconstructed with autologous sural nerve grafts. In four cases, a reconstructive procedure was intraoperatively abandoned as a result of the large extent of the lesion. RESULTS: Thirty-six patients with an adequate follow-up period of at least 18 months were included in this study. Among those with external neurolysis, 73% (eight out of 11) showed a good functional outcome, obviating the need for a kick up foot brace (M >or= 4). In the interfascicular neurolysis group, 71% (five out of seven) exhibited a similar outcome. In the grafted group, however, only 28% (five out of 18) obtained a functionally useful result dependent on graft length. A graft length under 6 cm led to a functionally useful outcome in 44% of patients (four out of nine) compared with 11% (one out of nine) when the graft length was greater than or equal to 6 cm. In six patients, muscle-tendon transfers were performed, resulting in strong, useful foot lift. CONCLUSION: Peroneal nerve lesions lacking regenerative signs should be explored. A functionally useful result (M >or=4) was achieved in 72% of the patients with either external or internal neurolysis and in 28% of the patients after a nerve graft procedure. Patients in whom nerve surgery failed to reconstitute useful foot lift need to be evaluated for their suitability to undergo a tendon transfer procedure. PMID- 18425014 TI - Cervical spondylotic myelopathy treated by oblique corpectomy: a prospective study. AB - OBJECTIVE: Anterolateral partial oblique corpectomy (OC) aims to decompress the cervical spinal cord without subsequent fusion and saves the patient from graft-, instrument-, and fusion-related complications. Although it is a promising technique, there are few studies dealing with its efficacy and safety. METHODS: In this prospective study, 40 consecutive patients underwent an OC (one to four levels from C3 to C7) for cervical spondylotic myelopathy; they ranged in age from 43 to 78 years (mean, 55 yr). The average follow-up period was 59 months (range, 24-98 mo). Clinical and radiological data were analyzed to assess the results and find possible factors related to outcomes. RESULTS: Thirty-seven (92.5%) of the 40 patients improved by the 6-month follow-up examination according to the Japanese Orthopedic Association score. The improvement was the most prominent in lower extremity dysfunction. Recovery was positively correlated with the preoperative Japanese Orthopedic Association score (r = 0.37, P = 0.018). Permanent Horner's syndrome developed in four patients (10%). During the long-term follow-up period, neurological improvement was maintained and there were no signs of postoperative instability, posture change, or axial pain. CONCLUSION: OC for treating multilevel cervical spondylotic myelopathy achieved good results with a low morbidity rate. The results of the current study suggest that OC is a good alternative to conventional median corpectomy and fusion techniques in selected cases. PMID- 18425015 TI - Predictors of ambulatory function after decompressive surgery for metastatic epidural spinal cord compression. AB - OBJECTIVE: Metastatic epidural spinal cord compression (MESCC) is a relatively common and debilitating complication of metastatic disease that often results in neurological deficits. This study was designed to explore associations with maintaining and regaining ambulatory function after decompressive surgery for MESCC. METHODS: Seventy-eight patients undergoing decompressive surgery for MESCC at an academic tertiary care institution between 1995 and 2005 were retrospectively reviewed. Fisher's exact analysis was used to compare preoperative ambulatory and nonambulatory patients. Multivariate Cox proportional hazards regression was used to identify associations with either maintaining or regaining the ability to walk. RESULTS: Patients were followed for 7.1 +/- 1.6 (mean +/- standard deviation) months after surgery. Preoperative nonambulatory patients required more extensive surgery (increased operative spinal levels and number of laminectomies) and had more surgical site complications (wound dehiscences and cerebrospinal fluid leaks) compared with preoperative ambulatory patients. From the multivariate analysis, preoperative ability to walk (relative risk [RR], 2.320; 95% confidence interval [CI], 1.301-4.416; P < 0.01) independently increased the likelihood of ambulation at the last follow-up evaluation 2.3-fold. Pathological vertebral compression fracture at presentation (RR, 0.471; 95% CI, 0.235-0.864; P = 0.01) independently decreased the likelihood of ambulation at the time of the last follow-up evaluation 2.1-fold. For patients unable to walk at the time of surgery, preoperative radiation therapy (RR, 0.406; 95% CI, 0.124-0.927; P = 0.03) decreased the likelihood of regaining the ability to walk 2.5-fold. Symptoms present for less than 48 hours (RR, 2.925; 95% CI, 1.133-2.925; P = 0.02) and postoperative radiotherapy (RR, 2.595; 95% CI, 1.039 8.796; P = 0.04) independently increased the likelihood of regaining ambulatory ability 2.9- and 2.6-fold, respectively, by the time of last follow-up evaluation. CONCLUSION: The identification of these associations with neurological outcome may help guide in the preservation or return of ambulation after surgery for patients with MESCC. PMID- 18425016 TI - The impact of minimally invasive spine surgery on perioperative complications in overweight or obese patients. AB - OBJECTIVE: Open lumbar spinal surgery in overweight or obese patients has been associated with increased risk of perioperative complications. The impact of minimally invasive spinal (MIS) surgery on the incidence of perioperative adverse events in overweight or obese patients, however, has not been well evaluated. METHODS: A retrospective review of consecutive patients undergoing lumbar MIS surgery from January 2006 to April 2007 was performed. Of the 77 patients identified, 56 had a body mass index (BMI) of 25.0 kg/m2 or greater. RESULTS: Of the 56 patients with a BMI of 25 kg/m2 or greater, 32 (57.1%) were men; the mean age was 54.1 years. The mean BMI was 31.0 kg/m2 (range, 25.1-43.8 kg/m2). Using a broad definition of an adverse event, eight (14.3%) complications were identified. In the discectomy/laminotomy subgroup (31 patients), two (6.5%) adverse events were noted. In the fusion subgroup (25 patients), six (24%) adverse events were noted, most of which were minor. Of the 21 patients with a BMI less than 25 kg/m2, eight (38.1%) were men, and the mean age was 43.7 years. The mean BMI was 22.5 kg/m2 (range, 16.8-24.6 kg/m2). Three (14.3%) complications were noted overall. In the discectomy/laminotomy subgroup (17 patients), two (11.8%) adverse events occurred. One (25%) complication developed in the four patients making up the fusion subgroup. There was no statistically significant difference in complication rates between groups. Logistic regression also found no statistically significant relationship between BMI and perioperative complications. CONCLUSION: There does not appear to be an increased risk of developing perioperative complications in overweight or obese patients undergoing MIS surgery, which may reflect a potential benefit of the MIS approach. PMID- 18425017 TI - Neurosurgical emergency transfers to academic centers in Cook County: a prospective multicenter study. AB - OBJECTIVE: The absence of surgical subspecialty emergency care in the United States is a growing public health concern. Neurosurgery is a field lacking coverage in many areas of the country; however, this is generally thought to be of greater concern in rural areas. Because of decreasing numbers of neurosurgeons, medical malpractice, and liability concerns, neurosurgery coverage is becoming a public health crisis in urban areas. Our objective was to quantify neurosurgical emergency transfers to academic medical centers in Cook County, IL, including patient demographics, reasons for transfer, time lapse in transfer, and effects on patient condition. METHODS: Data on neurosurgery emergency transfers was gathered prospectively by all five of the academic neurosurgery departments in Cook County, IL, over a 2-month period. Patient demographics devoid of identifiers, diagnosis, transfer origin, time lapse of transfer, and patient condition at the time of transfer and at the receiving hospital were recorded. RESULTS: Two-hundred thirty emergent neurosurgical transfers occurred during the study period. The most common diagnoses were parenchymal intracerebral hemorrhage (33%) and subarachnoid hemorrhage (28%). Sixty-six percent of neurosurgical transfers to academic medical facilities originated at hospitals without full time neurosurgery coverage. The mean time to transfer for all patients was 5 hours 10 minutes (standard deviation, 3 h 42 min; range, 1-20 h 12 min). A decline in Glasgow Coma Scale score was seen in 29 patients. A shortage of neurosurgical intensive care unit beds occurred on 55% of the days in the study. Only 19% of the emergency cases were related to cranial trauma, and only 3% of transfers came from Level 1 trauma centers. CONCLUSION: A combination of factors has led to decreases in availability of neurosurgical coverage in Cook County community hospital emergency departments. This has placed an increased burden on neurosurgical departments at academic centers, and, in some cases, delays led to a decline in patient condition. Eighty-one percent of the cases were not related to cranial trauma; thus, acute care trauma surgeons would be of little use. Coordinated efforts among local governments, medical centers, and emergency medical services to regionalize subspecialty services will be necessary to manage this problem. PMID- 18425018 TI - Microsurgical anatomy of the diaphragma sellae and its role in directing the pattern of growth of pituitary adenomas. AB - OBJECTIVE: To evaluate the anatomic aspects of the diaphragma sellae and its potential role in directing the growth of a pituitary adenoma. METHODS: Twenty cadaveric heads were dissected and measurements were taken at the level of the diaphragma sellae. RESULTS: The diaphragma sellae is composed of two layers of dura mater. There is a remarkable variation in the morphology of the diaphragm opening. The average anteroposterior distance of the opening was 7.26 mm (range, 3.4-10.7 mm) and the average lateral-to-lateral distance was 7.33 mm (range, 2.8 14.1 mm). CONCLUSION: The variability in the diameter of the opening of the diaphragma sellae could explain the growth of pituitary tumors toward the cavernous sinus or toward the suprasellar region. PMID- 18425019 TI - Clinical problem solving: lightning never strikes twice? PMID- 18425020 TI - Costanzo Varolio (Constantius Varolius 1543-1575) and the Pons Varolli. AB - COSTANZO VAROLIO (CONSTANTIUS Varolius) (1543-1575) was born in Bologna and died in Rome. This professor of anatomy and papal physician was the first to examine the brain from its base up, in contrast with previous dissections of this organ performed from the top down. Varolio was the first to describe many structures, including the pons, which is still known today as the pons Varolli. Varolio was a pupil to the well-known anatomist Aranzio, who was in turn a pupil of Vesalius. Our current understanding of the nervous system is based on the early anatomic descriptions and depictions by such individuals as Varolio. PMID- 18425022 TI - Quality Management in Health Care. From the editor. PMID- 18425021 TI - An account of the life and achievements of Miss Diana Beck, neurosurgeon (1902 1956). AB - DIANA BECK (1902-1956) WAS probably the first female neurosurgeon in the world and played a large part in establishing neurosurgery in the United Kingdom. She served as the consultant advisor in neurosurgery for the Emergency Medical Service in the southwest region during the Second World War and subsequently set up the neurosurgical unit at Frenchay Hospital in Bristol before becoming a consultant at The Middlesex Hospital in London in 1947. There, she was the first woman to be given charge of men in a consultant capacity in a major London teaching hospital. During her 31-year career, she was the only female neurosurgeon in Western Europe and the United States. Miss Beck came to the attention of the general public when she operated on Alan Alexander Milne in 1952. However, she was better known in neurosurgical circles for her interest in intracranial hemorrhage. In this article, some of her original research, case reports, and her seminal paper on the surgical treatment of intracerebral hemorrhage are presented. They are a stark reminder of the limited investigations available at that time. In addition to her neurosurgical achievements, Miss Beck was a remarkable character and teacher. Her Saturday ward rounds proved so popular that they competed successfully with the students' weekend plans. In a specialty in which women continue to represent a disproportionately small percentage of the workforce, Miss Beck was one of our earliest role models. PMID- 18425023 TI - Pay for performance and medical education: strategies for preparing physicians of the future. AB - Performance-based reimbursement has become an increasingly important topic in the field of medicine and one that has met with significant legislative support. Small- and large-scale pilot programs in the United States and more comprehensive programs implemented abroad have yielded preliminary findings that raise several interesting questions regarding the form that pay-for-performance programs will take and concerns about the unintended and unforeseen consequences of this new reimbursement approach. One important area that has not been explored, however, is the potential implications of pay for performance to "clinician educators" individuals from diverse health-related disciplines who both provide health care and are responsible for training the next generation of health care professionals. Because the effect of pay for performance is likely to vary by health care discipline, we focus here on potential implications for physician educators. Our objective is to analyze the experience to date with pay for performance, make predictions from these experiences about the potential impact of pay for performance on the education of resident physicians, and propose educational strategies that might be useful in positioning physician trainees for success as they enter the workforce. PMID- 18425024 TI - Pay for performance in hospitals: implications for nurses and nursing care. AB - As pay-for-performance programs gain momentum, hospital administrators and clinical leaders will need to consider the organization's infrastructure and measures that promote quality management initiatives. Many hospital performance measures by the Centers for Medicare & Medicaid Services involve chronic diseases that may be best managed by an interdisciplinary team-based approach, of which nurses are significant members. While the primary focus of pay-for-performance has been concentrated on physicians, comparatively less attention has been given to the potential impact on nurses and nursing care. Moreover, the impact of the pay-for-performance measures on nursing labor and processes has not been well studied. Within acute care settings, increasing attention has focused on the structure of nursing, such as number and skill mix of nursing personnel, processes of care, and influence on patient outcomes. As pay-for-performance standards evolve and encompass patient outcomes, attention to nursing's contribution will follow. Nursing leadership will need to address a number of strategies to (a) address the impact of pay-for-performance on nursing performance measures as well as (b) on staff nurses' ability to contribute to the organization's efforts in achieving pay-for-performance standards, including education, documentation, team collaboration, and patterns of care. PMID- 18425025 TI - Reducing wait time in a hospital pharmacy to promote customer service. AB - PURPOSE: The purpose of this study was to compare the effects of 2 different interventions on wait times at a hospital outpatient pharmacy: (1) giving feedback to employees about customer satisfaction with wait times and (2) giving a combined intervention package that included giving more specific feedback about actual wait times and goal setting for wait time reduction in addition to the customer satisfaction feedback. The relationship between customer satisfaction ratings and wait times was examined to determine whether wait times affected customer service satisfaction. SUBJECTS AND METHODS: Participants were 10 employees (4 pharmacists and 6 technicians) of an outpatient pharmacy. Wait times and customer satisfaction ratings were collected for "waiting customers." An ABCBA' within-subjects design was used to assess the effects of the interventions on both wait time and customer satisfaction, where A was the baseline (no feedback and no goal setting); B was the customer satisfaction feedback; C was the customer satisfaction feedback, the wait time feedback, and the goal setting for wait time reduction; and A' was a follow-up condition that was similar to the original baseline condition. RESULTS AND CONCLUSIONS: Wait times were reduced by approximately 20%, and there was concomitant increased shift in levels of customer satisfaction, as indicated by the correlation between these variables (r = -0.57 and P < .05). Given the current prescription-filling process, we do not expect that major, additional reductions in wait times could be produced. Many variables may account for the variability in any individual customer's wait time. Data from this study may provide useful preliminary benchmarking data for standard pharmacy wait times. PMID- 18425026 TI - Watching your wait: evidence-informed strategies for reducing health care wait times. AB - Nobody wants the health care system to be characterized by long, involuntary waits for treatment. Both a strong theoretical rationale and a growing number of case studies support approaches that address the root cause of long waits-usually a poorly designed system, rather than an absolute lack of capacity. This structured review of both peer-reviewed and "gray" literature on waitlist management identifies the 7 common problems that underlie long waitlists and describes remedies that have been successfully applied, using Canadian and British examples with international relevance. Understanding these issues can help administrators and providers develop effective wait-reduction strategies in diverse health care settings. PMID- 18425027 TI - Different time perspectives of the doctor and the patient reduce quality in health care. AB - Time-related problems interfere with treatment decisions and evaluations in health care, both from the perspective of the doctor and from the perspective of the patient. The compliance level of the patient and subsequent evaluations in clinical practice might be affected by a discrepancy in the time perspective. Context factors related to time and health perspectives are relevant to clinical decisions and quality management. A summary of evaluation factors in quality management is presented: (a) the time perspective of the patient is different from the time perspective of the doctor, both in an objective and in a subjective manner; (b) disease chronicity chronic affects the perception of time; (c) assessments need to extend over a period sufficiently long for variations in a disease activity to be noticed; (d) there is variation both in time for an outcome to occur and in time span for that outcome; (e) the number of patients benefiting from certain drugs and variability over time is valuable information; (f) the outcome of a specified treatment could be estimated for different periods in a sequence; and (g) changes occur in judgments and decisions over time both for the doctor and for the patient. PMID- 18425028 TI - How work context shapes physician approach to safety and error. AB - A study was undertaken to examine how different hospital clinical settings compare in their capacity for physicians to attend to safety and employ a learning approach to error. Multiple qualitative methods were used to examine medical residency teams in the emergency department, surgery department, and the medical intensive care unit. The focus was on how physicians responded to errors that occurred and safety-related issues, and what features of the surrounding work context were associated with those responses. Observations of 3 separate medical residency team's everyday work were conducted over three 3-week spans, and follow-up interviews with select team members were conducted after each observational period. Evidence supported that physician capacity for attending to safety and error is shaped by structural features of the surrounding work context within hospitals, as well as the cultural dynamics inherent in physician groups working in a given clinical setting. Compared with the emergency department and surgery, the medical intensive care unit offered the greatest potential from a work setting perspective to have heightened physician attention to safety and error. This was due to its more collegial environment, the low permeability of its doctors and patients, more available physician downtime and group interactions, fewer disruptions, and greater work predictability. The emergency department had less capacity because of its intense workload, excessive disruptions, and high unpredictability of the clinical work found there. Surgery's capacity to deal with error and safety issues was adversely affected by the emphasis on hierarchy among surgeons, the high permeability of surgeons across hospital work settings, emphasis on individual blame when mistakes occurred, and workload. These findings highlight the necessity for health care organizations to conduct regular assessments of their clinical environments to help identify the workplace factors that shape clinician approach to safety and error. It also calls into question the singular, uniform approaches to enhancing quality and safety within health care organizations. PMID- 18425029 TI - A parametric control chart adjustment for handling serial dependence in health care quality data. AB - Numerous tools are available to health care quality managers geared toward helping them make data-based inferences about quality processes. Recently, in this journal, Tukey's control chart technique was promoted as a good option for handling short streams of time series data when the assumption of data normality cannot be confirmed. Although this technique does not appear to perform well with serially dependent (or autocorrelated) data, an autocorrelation-corrected version of the technique is now available. However, when managers wish to capitalize on the superior power of parametric control charts (ie, when data sets are large and normality can be confirmed), there are currently few options available in the way of statistical process control techniques that appropriately handle autocorrelated data. In this article, the authors report the empirical false positive rates and power performance of the mean-sigma (X-S) control chart technique under various levels of autocorrelation. Results indicate that this popular technique offers poor false-positive control with autocorrelated data. Next, the authors describe a method for autocorrelation correction and finally compare the autocorrelation-corrected X-S chart with the original X-S technique. The autocorrelation-corrected X-S chart demonstrates better type I error control with similar power to the original chart and may offer quality managers an important tool for appropriately handling autocorrelated quality data. PMID- 18425030 TI - Patient safety and the integration of graduate nurses into effective organizational clinical risk management systems and processes: an Australian study. AB - OBJECTIVES: This article explores and describes key processes facilitating the integration of neophyte graduate nurses into safe and effective organizational systems and processes as part of a hospital's overall local clinical governance and clinical risk management (CRM) program. METHODS: This study was undertaken using an exploratory descriptive case study method. Four sample units of analysis were used, notably, 2 cohorts of graduate nurses (n = 11) undertaking a 12-month graduate nurse transition program; key stakeholders (n = 34), that is, nurse unit managers, clinical teachers, preceptors, a quality manager, a librarian, and senior nurse administrators employed by the participating health service; and pertinent literature sources. RESULTS: Data revealed that the cohort of graduate nurses under study became substantively integrated into the hospital's organizational and patient safety system processes within the first 3 to 4 months of their employment. Processes most contributing to this integration included the graduates receiving pertinent information about CRM at the very beginning of their employment (at induction) and thereafter throughout their graduate nurse year. In addition, the graduates needed focused support to assist their efforts to integrate the CRM systems and processes of their employer organization into their "normal working practices." CONCLUSIONS: In order for new graduate nurses to become integrated into safe and effective organizational systems and processes as part of a hospital's overall local clinical governance and CRM program, they needed pertinent information on CRM at the beginning of their employment and thereafter be supported throughout their graduate nurse year to develop and translate that information into their "routine" (everyday) practice. Further comparative research involving larger cohorts of graduate nurses is required. PMID- 18425031 TI - Development of communities of practice to facilitate quality improvement initiatives in surgical oncology. AB - BACKGROUND: The process of developing clinical guidelines and standards for cancer treatment and screening is well established in the Ontario health care system; however, the dissemination and implementation of such guidelines and standards are more recent undertakings. Traditional implementation strategies to improve surgical practice and the delivery of cancer care have not been consistently effective. There is a recognized need to develop integrated models that offer direct support for implementation strategies. Such a model should be feasible, adaptable, and open to evaluation across diverse surgical settings. DISCUSSION: Research suggests that successful implementation should consider tools and expertise from other disciplines. This article considers a community of practice (COP) model to provide a supportive infrastructure for quality improvements in cancer surgery. The COP model was adapted for cancer surgeons. It is supported by 5 enablers referred to as tools: communication system, project development support, access to data, access to evidence review, and accreditation with continued medical education and continued professional development. These tools need to be part of an infrastructure that is both provided and supported by a team of administrators and health care professionals, who have active roles and responsibilities. Therefore, the primary objective of this article is to describe our COP model in cancer surgery including the key success factors necessary for providing the infrastructure and tools. The secondary objective is to offer the integrated COP model as a basis for future research and the evaluation of various collaborative improvement projects. SUMMARY: Building on knowledge management concepts, we identified the 4 essential processes that should be targeted by implementation strategies. A common COP evaluation framework uses the outcomes of 4 knowledge conversion modes-organizational memory, social capital, innovation, and knowledge transfer-as proxies for actual provider and organizational behavior. Insights from different collaborative improvement projects described in a consistent way could inform future research and assist in the collation of systematic reviews on this topic. PMID- 18425032 TI - Personal quality improvement and individualized health process control: an update. AB - Chronic disease management is about personal quality improvement. This involves tracking personal health measures over time to develop the patient's understanding of common and special cause variation. This can be done through the use of run, control, and time between charts. This review gives examples of different methods taken from the literature. PMID- 18425035 TI - Transmission of donor diseases: efforts to protect recipients and increase deceased donor organ transplants. PMID- 18425036 TI - Workshop on cancer risk after solid organ transplantation. Guest editors' introduction. PMID- 18425037 TI - Quality and safety in the Italian donor evaluation process. AB - BACKGROUND: The shortage of available organs, has increasingly prompted the use of elderly donors, with a consequent growth of possible risk factors. In this context the risk of donor-recipient transmission of infectious or neoplastic pathologies may be considered as a major issue; in each case for each organ potentially available, acceptable quality must be provided and unacceptable risks must be avoided. METHODS: We are presenting here the process of risk management followed by the Italian centers. In 2001, the Italian National Transplant Centre created a national commission of experts, with the mission of defining guidelines for the evaluation process of the potential organ donor. As a supplement to these measures, the Italian National Transplant Centre has supported transplant network health workers through ad hoc developed information tools and an expert task force (second opinion) for evaluation of doubtful cases. RESULTS: Starting from the date of guidelines application and second opinion start up, 9519 potential cadaveric donors were reported in Italy. Of these, 1611 presented a neoplastic or infectious risk. Over this period, 4861 donors were used for transplantation, equal to 48.5% of reported donors. Among the 1611 donors, who had been diagnosed at risk, 674 were neoplastic-disease affected donors and 937 infection-disease affected donors. CONCLUSIONS: At the European level, several new activities have been recently implemented to increase organ safety. In Italy, new guidelines and actions to ensure organ safety have been implemented. The evaluation of the impact of these actions will be performed in the near future. PMID- 18425038 TI - The Italian multiorgan donor cancer screening protocol: 2002-2005 experience. AB - BACKGROUND: We describe the results of the application of the Italian donor cancer screening protocol to all the 7608 candidate multiorgan donors presented in Italy in 2002-2005. METHODS: All suspect findings raised in the two presurgical and surgical phases of the protocol were investigated by extemporary pathologic evaluation. Donors were classified as standard risk (no transmissible risk); nonstandard risk (low-risk of transmission, eligibility restricted to certified clinical emergencies pending informed consent); and unacceptable risk (unconditional exclusion because of high-risk pathologies). RESULTS: The protocol was successfully implemented for all 7608 candidates. In addition to 8 (0.1%) independent exclusions, clinical suspicion of cancer was raised for 337 (4.6%) donors. According to pathological examination 198 donors (2.6%) were judged at unacceptable risk of tumor transmission; 80 (1%) were included in the "standard risk". Used standard risk and nonstandard risk donors provided a total of 241 organs in 231 recipients. Although no suspect was raised after implementation of the protocol, a malignant tumor was discovered after organ transplantation in 14 (0.2%) donors. All the recipients transplanted with organs from ascertained nonstandard risk donors or from neoplastic donors who donated by accident have been carefully followed. At the time of most recent follow-up no donor/recipient tumor transmission has been reported. CONCLUSIONS: Implementation of the multiorgan cancer screening protocol is feasible at a national level in Italy. In view of the increasing demand for organs our protocol provides a useful tool for rationalization of the use of organs from neoplastic marginal donors. PMID- 18425039 TI - The Spanish National Transplant Organization (ONT) tumor registry. AB - BACKGROUND: It is important to know the actual risk of tumor transmission from donor to recipient and the serious consequences for the recipient. Tumor registries can help us to improve our knowledge about this problem. METHODS: We have studied all the donors registered in the Spanish National Transplant Organization from January 1, 1990, to December 31, 2006, and especially the donors with a malignant tumor as well as the recipients who have received an organ from these donors. RESULTS: We found 117 donors with a malignant tumor (5.8 per 1000 donors). One hundred fifty-five recipients were transplanted with an organ from these donors. The average age (SD) of donors with tumor was 53 (17) years. The most frequent cause of death was cerebral stroke in 81 donors. Donors with tumor are older than donors without tumor. The cause of death was cerebral stroke more frequently in donors with tumor than donors without tumor. Twenty-two of the recipients who received an organ from a donor with a tumor are dead. In 7 of these 22 recipients the death was cancer-related. Only 13 of the 100 recipients studied developed a malignant tumor, and only 10 of these tumors were donor-related. CONCLUSIONS: The profile of a donor who could have a tumor was most frequently an elderly person who had died of a cerebral stroke. In our experience, the risk of tumor transmission from donors to recipients is low and depends on the aggressiveness of the donor tumor. PMID- 18425040 TI - Gender differences in survival after lung transplant: implications for cancer etiology. AB - Predictors of survival after lung transplant were analyzed in a large cohort of 990 lung transplanted patients from a single center. The overall survival was 41.6%, (41.5% in males, and 41.8% in females), the average length of the follow up was 45.84+/-51.98 months (range 0-282.47 months). Females tend to live longer than males: 50.75+/-55.41 months versus 40.64+/-47.60 months, respectively. Males had a risk of dying during the follow up that was 1.18 (95% CI 1.01-1.40) relative to females, after adjusting for ethnicity, age, smoking status, diagnosis and donor characteristics. Females who had at least one full term pregnancy during their life had better survival rates than females who had no full term pregnancies. Our results of a better survival after lung transplant in females, and among them in those who had at least a full term pregnancy support the hypothesis of a hormonal contribution to survival and of the development of immunotolerance after pregnancy. This model could be useful for understanding the role of immunity in cancer development. PMID- 18425041 TI - Malignancy in lung transplantation: biomarkers, gender differences, and consideration of a systems biology approach. AB - We present here new molecular tools such as gene expression microarrays as promising biomarkers for diagnosis and prognosis in lung cancer as well as for an early diagnosis of malignancy after lung transplantation. Data regarding gender differences in survival, recently collected in a single-institution study, may allow better targeting of immunosuppression goals to maintain effectiveness while minimizing cancer development and progression. Combining these differing types of data using an integrative approach that is the central theme of systems biology may allow us to provide powerful and more individualized risks of cancer after lung transplantation. PMID- 18425042 TI - Sebaceous epithelial-myoepithelial carcinoma of the salivary gland: clinicopathologic and immunohistochemical analysis of 6 cases of a new histologic variant. AB - Epithelial-myoepithelial carcinoma (EMC) of the salivary glands is an uncommon, low-grade malignant tumor. A recent report demonstrates sebaceous differentiation in this tumor even though its significance has never been documented as a precise histologic variant. Six cases of EMC exhibiting sebaceous differentiation (sebaceous EMC) of the parotid gland were analyzed for their clinicopathologic features and immunohistochemical characteristics. In addition, primary salivary sebaceous carcinomas were also examined for comparison. In our series, the incidence of sebaceous EMC was 0.2% among 3012 cases of parotid gland tumors and 14.3% of all EMC cases. The 6 patients comprised 2 men and 4 women, age ranging from 77 to 93 years (mean, 83.7 y). Neither cervical lymph node nor distant organ metastases were found in any cases of sebaceous EMC and no patients died of disease, though local recurrences developed in 1 patient. Conversely, cervical lymph node metastasis was detected in 2 of 3 patients with sebaceous carcinoma, 1 of whom died of disease at 12 months. Histologically, all 6 tumors had an area of sebaceous differentiation admixed with features of bilayered ductal structures typical of EMC. A component of sebaceous differentiation was distributed diffusely in 4 tumors and focally in 2. Cytologic atypia of sebaceous EMCs was lesser than that of sebaceous carcinomas. Immunohistochemically, putative myoepithelial markers such as alpha-smooth muscle actin, calponin, p63, cytokeratin 14, S-100 protein, and vimentin were highly expressed in sebaceous EMC. However, the expression of the latter 4 markers was also observed in primary sebaceous carcinomas, whereas these tumors were all negative for alpha-smooth muscle actin and calponin. Positive immunoreactivity for epithelial membrane antigen, adipophilin, and perilipin confirmed sebaceous differentiation in EMC. These results indicate that sebaceous EMC is a low-grade malignancy, similar to conventional EMC. Our data also suggest that immunohistochemical examination of specific myoepithelial markers is helpful in distinguishing sebaceous EMC from sebaceous carcinoma, which may occasionally be associated with an aggressive clinical course. PMID- 18425043 TI - Benign nerve sheath tumors on urinary bladder biopsy. AB - Benign nerve sheath tumors (schwannoma and neurofibroma) involving the urinary bladders are rare with only case reports and limited series. We identified 6 neurofibromas and 2 schwannomas involving the urinary bladder. Of the 8 cases, 7 were sent to one of the authors in consultation and the last came for treatment to The Johns Hopkins Hospital. Patients (3 males, 5 females) ranged in age from 3 to 69 years old (average 36 y old). Presenting symptoms included recurrent infection in 6 patients, hematuria in 3 patients, and irritative symptoms in 3 patients (some patients with 2 symptoms). Three patients had a pelvic mass on physical examination. Three patients (3/8) had a solitary lesion in the bladder, 4 patients (4/8) had multifocal lesions, and 1 patient had no information available. Five patients underwent local resection. One patient, who was 3 years old, had multiple plexiform neurofibromas within the bladder and did not have surgical treatment and another patient is also undergoing surveillance. One patient had no treatment information available. Seven patients had clinical follow-up information available. The length of follow-up varied from 2 to 124 months (average 47 mo). Three out of the seven patients with follow-up information had neurofibromas in other sites, including the skin, uterus, mesentery, and ureter. One patient was known to have other stigmata of neurofibromatosis. Both cases with schwannoma had only bladder involvement. No family history of neurofibromatosis and no genetic studies were performed on any of the patients. Three neurofibromas were of the diffuse type, with 2 cases plexiform, and 1 case could not be subtyped. Neither recurrence nor malignant transformation was demonstrated on follow-up. The importance of accurately diagnosing plexiform neurofibromas of the bladder is that a subset harbors neurofibromatosis. If the lesion is focal, conservative excision is the choice of treatment with a low risk of recurrence. Diffuse neurofibromas can be difficult to diagnose leading to delay of treatment and potentially the need for a more extensive excision. Once recognized as a neurofibroma, it is important to identify it as a diffuse neurofibroma, given its lack of relationship to neurofibromatosis. PMID- 18425044 TI - BD ProEx C: a sensitive and specific marker of HPV-associated squamous lesions of the cervix. AB - BD ProEx C (ProEx C) is a recently developed immunocytochemical assay that targets the expression of topoisomerase II-alpha and minichromosome maintenance protein-2, 2 genes shown to be overexpressed in cervical cancers. Recent studies validated this reagent in liquid-based cytology specimens and suggested its usefulness as an adjunct in the diagnosis of challenging cases. Limited information is available on its expression in tissue sections. This study aims to assess ProEx C expression patterns in benign, atypical, and dysplastic lesions of the cervix and to compare these patterns with p16 and Ki67 expression and with the presence of human papilloma virus DNA as determined by in situ hybridization. ProEx C positivity was limited to the basal layers of the epithelium in 75% of benign cervices. In the remaining 25%, staining extended into the lower half of the epithelium particularly in areas of squamous metaplasia and immature squamous metaplasia. In 92% of high-grade dysplasias [cervical intraepithelial neoplasia (CIN) II and III] strong positive staining for ProEx C involved the lower and upper halves of the epithelium. Condylomas and CIN I showed greater variability in staining pattern with ProEx C positivity extending into the upper half of the epithelium in 48% of cases. Statistically significant correlations were noted with presence of human papilloma virus DNA and with p16/Ki67 expression. Atypical squamous metaplasia showed varied staining with 24% being positive. To summarize, ProEx C is a reliable marker for high-grade CIN that can be applied to tissue sections to confirm the diagnosis of high-grade CIN and to triage cases of atypical squamous metaplasia. ProEx C may also have a potential role in triaging patients with low-grade CIN. ProEx C expression patterns are similar to those for p16 and Ki67 with most discrepancies occurring in the CIN I category. PMID- 18425045 TI - Intravascular large T-cell or NK-cell lymphoma: a rare variant of intravascular large cell lymphoma with frequent cytotoxic phenotype and association with Epstein-Barr virus infection. AB - Most cases of intravascular large cell lymphoma have a B-cell phenotype, but rare T-cell and natural killer (NK)-cell variants have been reported. We describe the clinicopathologic features of 4 patients (M:F=3:1; age range: 63 to 87; median age: 65) with intravascular large NK/T-cell lymphoma. The skin was the site of presentation in all patients (leg: 1 case; trunk: 1 case; trunk and extremities: 2 cases). Two patients had lesions confined to the skin; in 1 case concomitant involvement of the brain was detected and in 1 case no further studies were carried out. Immunohistology showed positivity for cytotoxic markers in 3/4 cases. One case had an NK phenotype similar to NK/T-cell lymphoma, nasal-type, whereas the other cases could not be precisely classified into specific categories (peripheral T-cell lymphoma, NOS). One of these cases was negative for cytotoxic markers and was positive only for CD2 and CD3 epsilon. Association with Epstein-Barr virus (EBV) was demonstrated in 2 cases by in situ hybridization, whereas 1 case was negative. All our patients had aggressive disease and died between 2 weeks and 7 months from presentation. Analysis of our cases and of those published in the literature shows that intravascular large NK/T-cell lymphoma is a rare, aggressive lymphoma with variable phenotypic features, frequent expression of cytotoxic proteins, true NK-cell phenotype and association with Epstein-Barr virus infection, and common presentation in the skin. Homogeneous studies on larger number of patients and reevaluation of cases published with incomplete phenotypic data would be necessary to gather more information on this extremely rare type of lymphoma. PMID- 18425046 TI - Expression of CD56 and WT1 in ovarian stroma and ovarian stromal tumors. AB - The immunophenotype of ovarian stroma and spindle cell tumors derived from ovarian stroma has not been well studied. We studied the expression of CD56, WT1, estrogen receptor-beta (ER-beta), progesterone receptor (PR), smooth muscle actin, S-100, CD34, and muscle specific actin in 16 normal ovaries, 17 ovarian fibromas, 11 ovarian cellular fibromas, 10 ovarian fibrothecomas, and 11 ovarian leiomyomas. In addition, we studied CD56 and WT1 expression in 7 cases of normal endometrium, 8 uterine smooth muscle tumors, 5 endometrial stromal tumors and 64 nongynecologic (GYN) spindle cell sarcomas. All normal ovaries, ovarian fibromas, fibrothecomas, and ovarian leiomyomas were positive for CD56 and WT1. Most of the normal ovaries, ovarian fibromas, ovarian fibrothecomas, and ovarian leiomyomata also expressed ER-beta and PR. Eight of 17 ovarian fibromas, 5 of 11 ovarian cellular fibromas, and 4 of 10 ovarian fibrothecoma with focal fibroblastic differentiation were positive for smooth muscle actin. A few cases of these tumors also expressed S-100 and CD34. Only rare cases of non-GYN spindle cell sarcomas expressed WT1. Our study results show that ovarian fibromas, fibrothecomas, and leiomyomas have a similar immunophenotype (positive for CD56, WT1, ER-beta, and PR) to that of ovarian stromal cells, supporting an ovarian stromal origin for these neoplasms. However, unlike normal ovarian stromal cells, ovarian fibromas, fibrothecomas, and leiomyomas can also show fibroblastic, smooth muscle, Schwannian, and solitary fibrous tumorlike differentiation. WT1 is a fairly specific marker for spindle cell tumors of gynecologic organs, including ovarian spindle cell tumors, endometrial stromal tumors, and uterine smooth muscle tumors. Non-GYN spindle cell sarcomas rarely express WT1. CD56 is strongly expressed in ovarian stromal cells but not in endometrial stromal cells. CD56 is often expressed by a wide variety of spindle cell sarcomas, thus, it has no value in differentiating GYN from non-GYN spindle cell tumors. PMID- 18425047 TI - Pneumolabyrinth without temporal bone fracture: different outcomes for hearing recovery. AB - Pneumolabyrinth is a rare clinical manifestation seen in the practice of otology. It occurs usually after head trauma with temporal bone fracture. However, pneumolabyrinth without temporal bone fracture is a very rare occurrence. When it occurs, it is usually diagnosed by high-resolution computed tomography of the temporal bone. Therapy for pneumolabyrinth is conservative treatment or an exploratory tympanotomy. Recently, we saw two cases of pneumolabyrinth after trauma without temporal bone fracture. The patients were treated with conservative management for the pneumolabyrinth. However, the degree of recovery from hearing loss was different for each patient. PMID- 18425048 TI - Endoscopic treatment of subcondylar fractures. AB - OBJECTIVES/HYPOTHESIS: To evaluate the effectiveness of endoscopic treatment of subcondylar fractures, highlighting an improved technique of repair that facilitates ease of repair. STUDY DESIGN: A retrospective review. METHODS: All subcondylar fractures treated by the author with the improved endoscopic technique from 2001 to 2007 were reviewed. RESULTS: A total 34 subcondylar fractures were initially treated with the outlined technique. Thirty-three of 34 were successfully managed with the endoscopic technique alone. There were no instances of facial nerve paralysis or palsy noted. There were two instances of malocclusion that were believed to be minor in the 27 of 34 patients who made themselves available for 6 week follow-up. Both of these patients had associated multiple maxillofacial fractures repaired. Average operative time from ramus incision start to completion of plate fixation for the subcondylar fracture was 32 (range, 21-49) minutes. CONCLUSIONS: The outlined technique results in improved ease of rigid endoscopic fixation of subcondylar fractures in the majority of patients. PMID- 18425049 TI - Relationship between rhinitis duration and response to nasal decongestion test. AB - OBJECTIVES/HYPOTHESIS: Nasal obstruction depends on allergic inflammation. Decongestion tests evaluate the reversibility of nasal airflow limitation. It has been previously reported that duration of persistent allergic rhinitis (PER) may involve important functional consequences. The purpose of the study was to evaluate the impact of the duration of rhinitis on the response to nasal decongestion test in a cohort of patients with PER. METHODS: A total of 312 patients with moderate-severe PER were prospectively and consecutively evaluated: 234 males and 78 females, mean age 23.6 years. A detailed clinical history was taken and complete physical examination, nasal endoscopy, skin prick test, rhinomanometry, and nasal decongestion test were performed for all patients. RESULTS: A strong inverse correlation was observed (Pearson's r = -0.81) between rhinitis duration (years) and posttest percentage change of nasal airflow values. CONCLUSIONS: The duration of PER may induce a progressive impairment of the response to nasal decongestion test. PMID- 18425050 TI - Secreting jugulotympanic paraganglioma with venous involvement into the thorax. AB - Paragangliomas are highly vascular tumors that arise from chief cells in extra adrenal paraganglia of the autonomic nervous system. Jugulotympanic paragangliomas involve the middle ear and jugular fossa. Secreting jugulotympanic paragangliomas with thoracic extension are rare. We report a case with unexpected involvement of the internal jugular vein. Otolaryngologists should be aware that the jugulotympanic paragangliomas traverse variable anatomic pathways. Multiple imaging options, including magnetic resonance venography, are useful and should include the neck for definitive characterization of tumor anatomy. Patients with refractory hypertension and masses suspicious for paraganglioma should be examined for functional tumors. PMID- 18425051 TI - Effects of long-term denervation on the rat thyroarytenoid muscle. AB - OBJECTIVES/HYPOTHESIS: To determine the effects of long-term denervation on the rat thyroarytenoid (TA) muscle and neuromuscular junctions. STUDY DESIGN: A quantitative histologic assessment of the TA muscle after long-term denervation. METHODS: Thirty Wistar rats were euthanized 10, 18, 26, 42, and 58 weeks after left recurrent laryngeal nerve resection. The areas of the entire muscle and individual muscle fibers were evaluated using hematoxylin-eosin staining, and neuromuscular junctions were detected by immunohistochemistry. Changes after denervation were evaluated by comparing the treated (T) and untreated (U) sides (T/U ratio). The ratio of the number of nerve terminals (NTs) to that of acetylcholine receptors (AChRs) (NT/AChR ratio) was also assessed. RESULTS: The average T/U ratio for the entire muscle area of the denervation groups ranged between 61.1% and 72.5% and did not differ significantly. Similarly, the T/U ratios for the individual muscle fiber area ranged between 45.0% and 51.9%, and the differences were not significant. The T/U ratio of AChRs at 58 weeks (35.3 +/ 20.2%) was significantly lower than that at 10 weeks (76.3 +/- 9.0%; P < .01). The NT/AChR ratios ranged between 30.3% and 35.6% and did not differ significantly among the denervation groups. CONCLUSIONS: The entire TA muscle area, individual muscle fiber area, and NT/AChR ratio did not decrease with long term denervation. Thus, the TA muscle may retain an ability to receive regenerating nerve axons. However, the ability of the TA muscle to receive nerve axons may deteriorate after an excessively long denervation period because the T/U ratio of AChRs decreased with long-term denervation. PMID- 18425052 TI - In reference to cost-effective diagnosis of ingested foreign bodies. PMID- 18425053 TI - In reference to bacteriological comparison of tonsil core in recurrent tonsillitis and tonsillar hypertrophy. PMID- 18425054 TI - Which statin? PMID- 18425055 TI - Diclofenac gel for osteoarthritis. PMID- 18425057 TI - Imaging neuroendocrine tumours with radionuclide techniques. AB - Neuroendocrine tumours (NETs) are a rare type of tumour that can range from well differentiated slow growing tumours to poorly differentiated aggressive tumours. NETs can arise in different parts of the body. Patients often present with a large spectrum of clinical syndromes, which depend on the presence of hormones secreted by the tumour type or by a mass effect. Several imaging modalities are used in the diagnosis of NETs. Radionuclide imaging plays an important role in diagnosis, staging and treatment of these tumours. Several radiopharmaceuticals are used to image NETs and are based on different cellular uptake mechanisms. This review will focus on the functional imaging of NETs using single photon emission computed tomography and positron emission tomography radiopharmaceuticals. PMID- 18425058 TI - Alterations of the tear film and ocular surface health in chronic smokers. AB - OBJECTIVE: To investigate the effects of chronic smoking on ocular surface and tear functions. METHODS: Fifteen right eyes of 15 healthy chronic smokers (9 men, 6 women; age range: 36-47 years) who smoked 20 cigarettes per day for 20 years and 20 eyes of 20 control non-smokers (12 men, 8 women; age range: 38-43 years) were included in this prospective study. All subjects underwent measurements of breath and haemoglobin CO concentration, tear lipid layer interferometry, evaporimetry, tear film break-up time (TBUT), Schirmer's I test, corneal fluorescein staining, conjunctival impression, and brush cytology. RESULTS: The mean Hb CO level was significantly higher in smokers compared to non-smokers. TBUT was also significantly shorter in smokers. Tear lipid layer showed significant slowing in spread over the tear film with a concomitant significant increase in tear evaporation rate. Conjunctival impression cytology revealed significant loss of goblet cells and squamous metaplasia in smokers. Brush cytology showed significant conjunctival neutrophil infiltration in smoker subjects. CONCLUSION: Chronic smoking induced distinctive quantitative and qualitative disturbances on the ocular surface health. PMID- 18425059 TI - Ocular manifestations in oculodentodigital dysplasia resulting from a heterozygous missense mutation (L113P) in GJA1 (connexin 43). AB - PURPOSE: To characterize the ophthalmic findings, intrafamilial variability, and molecular genetic basis of oculodentodigital dysplasia (ODDD; MIM no. 164200). METHODS: Ophthalmic examination included best-corrected visual acuity, slit-lamp biomicroscopy, direct and indirect ophthalmoscopy, Goldmann applanation tonometry and A-scan ultrasonography. Blood samples were taken for DNA extraction and mutation screening of GJA1 (connexin 43). RESULTS: All three affected individuals had characteristic features of ODDD. The ophthalmic features were epicanthus, microcornea, and the presence of glaucoma. The ocular phenotype resulted from a heterozygous T>C transition at nucleotide 338 in GJA1 (L113P) that was not detected in 120 chromosomes of unaffected individuals. The L113P mutation results in a nonconservative substitution in the cytoplasmic loop of Cx43 (GJA1) and is predicted to disrupt the high-order structure of Cx43. CONCLUSIONS: This report describes the ocular phenotype in a molecularly characterized ODDD syndrome family. The ocular features in this family highlight the key role Cx43 plays in eye development and in the development of glaucoma. L113P represents a pathogenic mutation in GJA1 (Cx43) and results in ODDD with marked intrafamilial variation in glaucoma type and severity. PMID- 18425060 TI - Vitreous management in penetrating trauma: primary repair and secondary intervention. AB - Ocular trauma involving the vitreous produces a very wide range of pathology, the treatment of which can confuse. In the past, the prognosis for vision following such injuries was poor. Modern closed intraocular surgical techniques have transformed the management of penetrating trauma and allowed salvage of vision in many previously hopeless cases. However, many controversies remain, including the timing of surgical intervention, the use of encircling buckles, and the type of internal tamponade. This article will discuss those controversies, and make some practical recommendations for the management of penetrating and perforating trauma involving the vitreous. PMID- 18425061 TI - Clinicopathological changes at the vitreoretinal junction: posterior vitreous detachment. AB - Separation of the vitreous and posterior hyaloid membrane (PHM) or posterior vitreous detachment (PVD) typically occurs between the ages of 45 and 65 years in the general population, but may occur earlier in myopic or otherwise predisposed individuals. Age-related synergetic changes occurring within the cortical and central gel must be distinguished from the PHM, which envelopes it. This study reports on the correlation between 'true' PVD seen clinically by the physician using dynamic examination, high-power slit-lamp biomicroscopy, and oblique illumination with some of its histological, immunohistochemical, and ultrastructural features post-mortem. The presence of the Weiss ring does not necessarily indicate total clean separation of PHM, nor does its absence confirm that the PHM remains attached, since it may be destroyed during the process of separation. Immediately prior to PVD with the vitreous gel attached, the PHM must, by definition, form part of the inner limiting membrane. The detached PHM frequently exhibits basement membrane (BM) and its indigenous laminocytes stain focally for GFAP and type IV collagen. The PHM is distinct from and much thicker than the BM of Muller cells alone and the factors that initiate or limit separation of the PHM require greater study, particularly the role of laminocyte proliferation and migration. PMID- 18425062 TI - Intravitreal pegaptanib sodium for refractory pseudophakic macular oedema. AB - PURPOSE: Evaluate the efficacy of intravitreal pegaptanib sodium (Macugen) in refractory pseudophakic cystoid macular oedema (CME). DESIGN AND METHODS: Prospective, nonrandomized, interventional case series. Four eyes of four patients with refractory pseudophakic CME to pars plana vitrectomy and intravitreal bevacizumab and triamcinolone, were treated with pegaptanib sodium, with a mean follow up of 4 months. Pre- and postinfection examinations included assessment of best-corrected visual acuity (BCVA) using the Early Treatment Diabetic Retinopathy Study chart (ETDRS), fluorescein angiography (FA), and optical coherence tomography (OCT). RESULTS: Visual acuity increased in all patients after intravitreal pegaptanib sodium. OCT showed improvement of the retinal thickness in the macular area. CONCLUSION: Intravitreal pegaptanib sodium (Macugen) is a promising treatment for pseudophakic cystoid macular oedema resistant to other medical treatment strategies. However, further study is needed to assess the treatment's long term efficacy and the need for retreatment. PMID- 18425063 TI - Pain response and follow-up of patients undergoing panretinal laser photocoagulation (PRP) with reduced exposure times. PMID- 18425064 TI - The prevalence of cystoid macular oedema on optical coherence tomography in retinitis pigmentosa patients without cystic changes on fundus examination. AB - PURPOSE: To determine the prevalence of cystoid macular oedema (CME) by optical coherence tomography (OCT) in retinitis pigmentosa (RP) patients with no evidence of cystic macular lesions on fundus examination. METHODS: We included 63 RP patients with no evidence of cystic-appearing macular changes on fundus examination. All patients underwent a complete ocular examination including best corrected visual acuity using an ETDRS (Early Treatment Diabetic Retinopathy Study) chart, intraocular pressure measurement, anterior segment examination, and a detailed fundus examination. On 50 of the 63 patients, Fourier-domain OCT was performed using the radial slicer protocol. An additional 13 of the 63 patients were scanned using the macular thickness protocol on a time-domain OCT unit. The diagnosis of CME was defined by the presence of hyporeflective lacunae with well defined boundaries on at least two of the scans. RESULTS: The mean age of patients included in the study was 36 years (range 9-71 years). Out of the 63 patients examined, 20 showed CME in at least one eye (32%), whereas 11 patients showed CME in both eyes (18%). CONCLUSIONS: Our findings demonstrate that a substantial number of RP patients with CME, as determined by OCT, may not show cystic changes by direct ophthalmoscopy or contact lens biomicroscopy. Knowledge of the high frequency for CME in such patients can serve to identify those who may be amenable to current or future treatment strategies of their macular oedema and can potentially impact on future therapeutic trials where visual acuity is used as an outcome measure. PMID- 18425065 TI - Correlation of previous acute angle-closure attack with extent of synechial angle closure in chronic primary angle-closure glaucoma patients. AB - AIM: To document any correlation between previous acute angle-closure attack and the extent of synechial angle closure in chronic primary angle-closure glaucoma (PACG) patients. METHODS: Consecutive cases of chronic PACG with patent peripheral iridotomy had gonioscopy performed. The extents of synechial angle closure of those chronic PACG eyes with previous documented acute angle-closure attack were compared to those eyes without such a history. RESULTS: A total of 102 chronic PACG eyes of 102 patients were recruited. Twenty-seven eyes (26.5%) had a previous documented acute angle closure, while 75 eyes (73.5%) did not. The mean extent of synechial angle closure +/-1 SD was 307+/-68 degrees (range, 150 360 degrees) in those chronic PACG eyes with a history of previous acute angle closure, compared to 266+/-89 degrees (range, 90-360 degrees) in those chronic PACG eyes without such a history (P=0.03, Student's t-test). There were no statistically significant differences between the two groups in age, LogMAR visual acuity, intraocular pressure (IOP), number of glaucoma eye drops, vertical cup-to-disk ratio, mean deviation or pattern SD in Humphrey automated perimetry, and anterior chamber depth (P>0.05). CONCLUSION: Previous acute angle-closure attack correlated with more extensive synechial angle closure in chronic PACG patients in this study. PMID- 18425066 TI - Myopathic ptosis following posterior sub-Tenon's triamcinolone acetonide injection. PMID- 18425067 TI - Prevalence of cystic macular lesions in patients with Usher II syndrome. AB - PURPOSE: To evaluate the prevalence of cystic macular lesions in patients with Usher II syndrome. METHODS: All Usher type II patients seen in the inherited eye disease clinic at the University of Illinois at Chicago between January 2002 and December 2007 were included (n=76). Each participating patient underwent a detailed clinical examination, including best-corrected visual acuity, slit-lamp biomicroscopy and dilated fundus examination. The presence of cystoid lesions was determined by optical coherence tomography (OCT), fundus fluorescein angiogram (FFA), fundus photographs and/or clinical examination. RESULTS: A cystic appearing macular change was observed in at least one eye in 19 out of the 76 patients (25%), 13 on the basis of OCT, five using FFA (two solely with the use of FFA and three based on clinical notes and FFA findings) and one based solely on clinical notes. Of the 18 patients with CME, determined by OCT or FFA, five (27.8%) showed either a funduscopically normal-appearing macula (n=4) or an atrophic appearing macular change (n=1). CONCLUSIONS: One-fourth of our total cohort of Usher II patients had cystic macular lesions. Moreover, a funduscopically normal-appearing macula was observed in 22% (n=4) of our 18 patients with cystic-appearing macular lesions on OCT and/or FFA testing. On the basis of the reasonably high prevalence of cystic macular lesions in our cohort, it would seem prudent to evaluate Usher II patients for the presence of cystoid macular oedema. PMID- 18425068 TI - Age-specific changes in the prevalence and management of optically correctable visual impairment between 1988 and 2000: the Ponza Eye Study. AB - PURPOSE: To obtain age-specific data on changes in the prevalence and management of optically correctable visual impairments (OCVIs) in Ponza, Italy. METHODS: Ophthalmologic examinations were carried out to 1000 Ponzans aged 40-87 years in 1988 and to 836 persons in 2000. Visual acuity (VA) was evaluated under uncorrected (VA(UC)), presenting (VA(PR)), and best-corrected (VA(BC)) conditions. We calculated the prevalence of total OCVIs (subjects with VA(UC)>0.5 logMAR and VA(BC)0.5 logMAR), and corrected OCVIs (VA(PR)15 letters. Mean OCT central foveal thickness (CFT) (n=13) was: baseline 325+/-123 microm, 12 weeks 343+/-130 microm, 24 weeks 321+/-115 microm; difference at 24 weeks was not statistically significant (P=0.9). A pigment epithelial detachment was present in 12 cases; height was reduced in 10 cases at 24 weeks. Persistent leakage on FA was seen in 13 out of 15 cases at 24 weeks. CONCLUSION: Early results of treatment of RAP with intravitreal pegaptanib suggest some stabilizing effect on this normally progressive disease. PMID- 18425071 TI - Antioxidant supplements to prevent or slow down the progression of AMD: a systematic review and meta-analysis. AB - INTRODUCTION: The aim of this review was to examine the evidence as to whether antioxidant vitamin or mineral supplements prevent the development of AMD or slow down its progression. METHODS: Randomised trials comparing antioxidant vitamin and/or mineral supplement to control were identified by systematic electronic searches (updated August 2007) and contact with investigators. Data were pooled after investigating clinical and statistical heterogeneity. RESULTS: There was no evidence that antioxidant (vitamin E or beta-carotene) supplementation prevented AMD. A total of 23 099 people were randomised in three trials with treatment duration of 4-12 years; pooled risk ratio=1.03 (95% CI, 0.74-1.43). There was evidence that antioxidant (beta-carotene, vitamin C, and vitamin E) and zinc supplementation slowed down the progression to advanced AMD and visual acuity loss in people with signs of the disease (adjusted odds ratio=0.68, 95% CI, 0.53 0.87 and 0.77, 95% CI, 0.62-0.96, respectively). The majority of people were randomised in one trial (AREDS, 3640 people randomised). There were seven other small trials (total randomised 525). CONCLUSIONS: Current evidence does not support the use of antioxidant vitamin supplements to prevent AMD. People with AMD, or early signs of the disease, may experience some benefit from taking supplements as used in the AREDS trial. Potential harms of high-dose antioxidant supplementation must be considered. These may include an increased risk of lung cancer in smokers (beta-carotene), heart failure in people with vascular disease or diabetes (vitamin E) and hospitalisation for genitourinary conditions (zinc). PMID- 18425073 TI - The availability and content of dental instrument manufacturers' decontamination information. AB - OBJECTIVE: The effective decontamination of reusable dental instruments is essential to reduce the risks from onward transmission of infectious diseases. There are therefore a number of legislative requirements placed upon manufacturers of medical devices (which includes dental instruments) to provide validated methods for the reprocessing of such devices. The aim of this study was to determine the availability and content of manufacturer's instructions for the reprocessing of reusable dental instruments. MATERIALS AND METHODS: A database of reusable dental instruments with details of their manufacturers was collated from information received from three dental hospitals. A questionnaire was sent to all the manufacturers requesting information about the reprocessing instructions for their products. The response from each manufacturer was assessed for the quality of the information and compliance with the British, European and International Standard, BS EN ISO 17664 (2004). RESULTS: The database from the three dental hospitals included over 800 items supplied by 54 different manufacturers/suppliers. Forty protocols were available for assessing compliance with BS EN ISO 17664 (2004). These protocols accounted for 25 (46%) manufacturers covering 300 devices. The majority (90%) of the returned questionnaires did not comply with the required standard and provided insufficient information to allow for the effective decontamination of the instruments. CONCLUSIONS: Manufacturers of medical devices are legally required to supply the user with validated instructions to enable effective decontamination of these devices. The information must be in a format as specified in BS EN ISO 17664 (2004). The information obtained in this survey demonstrated that the manufacturers' instructions fall short of the required regulatory requirements. The absence of such instructions increases the risk of cross-infection arising from inadequate cleaning, decontamination and sterilisation. PMID- 18425074 TI - Comparison of mineral trioxide aggregate and formocresol as pulp medicaments for pulpotomies in primary molars. AB - PURPOSE: The aim of this study was to compare the effect of white mineral trioxide aggregate (MTA) to that of formocresol (FC) as pulp dressing agents in pulpotomised primary molars. METHODS: In this clinical trial study, 60 lower second primary molars of 46 children were treated by a conventional pulpotomy technique. The teeth were randomly assigned to the MTA (experimental) and FC (control) groups by random numbered table. Following removal of the coronal pulp and haemostasis, the pulp stumps were covered with an MTA paste in the experimental group. In the control group, FC was placed with a cotton pellet over the pulp stumps. The teeth of both groups were restored with stainless steel crowns. Children arrived for clinical and radiographic follow-up evaluation after 6, 12 and 24 months. RESULTS: The treated teeth in FC group (n = 18) were clinically and radiographically successful after 24 months. The radiographic follow-up evaluation revealed one failure (furcation involvement) in 18 molars treated with MTA after 24 months. The treated teeth in MTA group were clinically successful 24 months postoperatively. Pulp canal obliteration was observed in one of the teeth treated with MTA and four of the teeth treated with FC. CONCLUSION: MTA could be used as a safe medicament for pulpotomy in cariously exposed primary molars and could be a substitute for FC. PMID- 18425072 TI - Targeting the glutamatergic system to develop novel, improved therapeutics for mood disorders. AB - Mood disorders are common, chronic, recurrent mental illnesses that affect the lives of millions of individuals worldwide. To date, the monoaminergic systems (serotonergic, noradrenergic and dopaminergic) in the brain have received the greatest attention in neurobiological studies of mood disorders, and most therapeutics target these systems. However, there is growing evidence that the glutamatergic system is central to the neurobiology and treatment of these disorders. Here, we review data supporting the involvement of the glutamatergic system in mood-disorder pathophysiology as well as the efficacy of glutamatergic agents in mood disorders. We also discuss exciting new prospects for the development of improved therapeutics for these devastating disorders. PMID- 18425075 TI - Evaluation of dentists' perceived needs regarding treatment of the anxious patient. AB - BACKGROUND: With regard to the management of dental anxiety in general dental practice, it has been considered that general dental practitioners (GDPs) are well placed to treat adults with mild forms of dental anxiety. However, little is known about the specific anxiety management techniques being used by GDPs in the UK. Aim To determine the views and experiences of dental practitioners in their current use of anxiety management techniques, their undergraduate and post graduation training in these techniques and future training needs. METHODS: A postal questionnaire was sent to a sample of GDPs working in the Midlands region (n = 750) in the UK. Dentists were randomly selected using lists provided by the primary care trusts for each locality. RESULTS: The response rate was 73% (n = 550). Of these, 90 were not included in the final analysis due to exclusion criteria set prior to questionnaire release. This left 460 questionnaires for analysis. Eighty-five percent of respondents agreed that dentists had a responsibility to help dentally anxious patients (n = 391). Dentists were asked their reasons for not using anxiety management techniques in practice. Psychological techniques, sedation (oral, inhalation, or intravenous) and hypnosis were reported as not having been used due to the paucity of time available in practice, a shortage of confidence in using these techniques and the lack of fees available under the NHS regulations. Also, 91% reported feeling stressed when treating anxious patients. When asked about the quality of teaching they had received (undergraduate and postgraduate), 65% considered that the teaching was less than adequate in the use of psychological methods, whereas 44% indicated that they would be interested in further training in psychological methods if financial support was available. CONCLUSION: The need for further training in managing the dentally anxious patient is supported by dentists' lack of confidence and inadequate training in treating such patients, as determined from the results of a postal questionnaire to UK GDPs. PMID- 18425076 TI - The role of the dental surgeon in detecting osteoporosis: the OSTEODENT study. AB - OBJECTIVE: To determine if thinning (<3 mm width) of the lower cortical border of the mandible on dental panoramic radiographs, as well as other clinical risk factors, may provide a useful diagnostic test for osteoporosis in young postmenopausal women. DESIGN: Six hundred and fifty-two subjects (age range 45-70 years) were involved in this multi-centre, cross-sectional study. SETTING: Patients were recruited from centres in Leuven (Belgium), Athens (Greece), Manchester (UK), and Malmo (Sweden). SUBJECTS AND METHODS: The subject's age, body weight, whether the patient took hormone replacement therapy or had a history of low trauma fracture were used to form a clinical osteoporosis risk assessment (the OSteoporosis Index of RISk or OSIRIS index). Each patient also received a dental panoramic radiographic examination. RESULTS: One hundred and forty subjects had osteoporosis involving at least one of the measurement sites (lumbar spine, femoral neck or total hip). Those with osteoporosis tended to have a low OSIRIS score and a thinned cortical mandibular border. The area under the ROC curve for using both cortical width and OSIRIS to predict osteoporosis was 0.90 (95% CI = 0.87 to 0.92). There was a significant improvement in the diagnostic ability of the combined OSIRIS and cortical width test over both tests applied separately (p <0.001). The cost effectiveness of the cortical width and OSIRIS model was improved by using a high specificity threshold rather than high sensitivity. However, this analysis ignores the costs associated with missed cases of osteoporosis. CONCLUSION: Dentists have a role to play in the detection and referral of patients at high risk of osteoporosis. PMID- 18425077 TI - Extrahepatic bile duct carcinoma with extensive intraepithelial spread: a clinicopathological study of 21 cases. AB - Extrahepatic bile duct carcinoma occasionally presents with intraepithelial spread for a considerable area around the main tumor. In this study, we compared clinicopathological features of extrahepatic bile duct carcinoma with and without extensive intraepithelial spread (>or=20 mm from the main tumor). Out of 117 cases of extrahepatic bile duct carcinoma, 21 (18%) were found to have extensive intraepithelial spread. Those cases were pathologically characterized by a papillary or nodular main tumor, a more differentiated histological grade, less deep invasion, and infrequent portal vein or hepatic invasion in comparison with cases without intraepithelial spread. Areas of intraepithelial spread histologically consisted of low-papillary growth (17 cases, 81%) and completely flat growth (4 cases, 19%) of carcinoma cells. The former histology corresponded to a macroscopic granular mucosa, whereas the latter growth was hardly detected by gross examination. Immunohistochemically, in 16 of 21 cases (76%), at least one of p53, CEA, and MUC1 was expressed in both the main tumor and the spreading area. Interestingly, patients with intraepithelial spread had a better postoperative prognosis than those without intraepithelial spread (P=0.009). However, three patients had anastomotic recurrence 54-130 months after surgery. In conclusion, intraepithelial-spreading bile duct carcinoma is characterized by papillary or nodular main lesions, a more differentiated histological grade, and less invasiveness. The presence of intraepithelial spread was not an indicator of a poor prognosis, but carcinoma in situ at the bile duct stump could cause late anastomotic recurrence after surgery. PMID- 18425078 TI - Expression of Smad2 and Smad4 in cervical cancer: absent nuclear Smad4 expression correlates with poor survival. AB - Alterations in transforming growth factor-beta signaling, due to a decrease in Smad2 and especially Smad4 expression, has primarily been reported in pancreatic and colorectal cancers, although loss of the chromosomal region 18q21.1, containing the loci of Smad2 and Smad4, is among the most frequent molecular alterations in cervical cancer. The aim of our study was to investigate whether decreased Smad2 and Smad4 protein expression in primary cervical cancers is associated with molecular alterations at 18q21.1, mutations in the functional domains of Smad2 and Smad4 or hypermethylation, and to assess the biological relevance of decreased Smad2 and Smad4 expression. Subsequently, Smad2, Smad4 and p21 protein expression was determined by immunohistochemistry in 117 primary cervical carcinomas, assembled in a tissue array. Smad signaling was shown to be associated with p21 mRNA expression. All the tumors expressed Smad2 or Smad4. Weak cytoplasmic Smad2 or weak cytoplasmic Smad4 expression could not be attributed to loss of heterozygosity at 18q21.1. Despite weak/moderate Smad2 expression and absent nuclear Smad4 expression, the coding regions of the functional MH1 and MH2 domains of Smad2 and Smad4 were unchanged, as assessed by sequence analysis. The Smad4 promoter region was unmethylated in tumor samples with weak/moderate cytoplasmic Smad4 expression. Remarkably, both weak cytoplasmic Smad4 expression and absent nuclear Smad4 expression significantly correlated with poor disease-free (P=0.003 and P=0.003, respectively) and overall 5-year survival (P=0.003 and P=0.010, respectively). Our findings support the hypothesis that Smad4 is a target molecule for functional inactivation in cervical cancer. PMID- 18425079 TI - Nuclear expression of survivin portends a poor prognosis in Merkel cell carcinoma. AB - Inhibition of apoptosis is a critical step in tumorigenesis in many cancers, including Merkel cell carcinoma; however, the exact regulatory mechanisms are not fully understood. Survivin is an inhibitor of apoptosis that is undetectable in most terminally differentiated normal human tissues, strongly expressed in embryonic and fetal organs and is strongly expressed in many different human cancers. In this study, we investigated the expression of survivin in cutaneous Merkel cell carcinoma using immunohistochemistry and correlated the findings with long-term clinical follow-up. We collected and immunostained 19 cases of Merkel cell carcinoma with antibodies to survivin. The median patient age was 79 years, with an average follow-up of 17 months, and a male/female ratio of 7:11. All but one sample represented primary lesions and two cases were obtained from one patient. Clinical follow-up was obtained in 15 cases (79%). All 19 cases of Merkel cell carcinoma demonstrated strong immunoreactivity for survivin. Survivin protein was localized and classified into predominately nuclear (N=8) or cytoplasmic (N=4) compartments. A mixed pattern of survivin expression was also seen in three cases. Cases with a nuclear staining pattern were distinguished by an aggressive clinical course, with seven of eight patients developing metastases or dead of disease on follow-up. Furthermore, all of the cases with predominately cytoplasmic survivin localization (N=4) were free of disease on follow-up. Merkel cell carcinomas represent aggressive malignancies regulated by apoptotic pathways. We demonstrate that survivin, a protein with a dual role in inhibition of apoptosis and regulation of cellular proliferation is expressed in Merkel cell carcinoma. Moreover, nuclear subcellular localization of survivin in Merkel cell carcinomas may portend a poor prognosis and identification of these cases may assist clinical management. PMID- 18425080 TI - Reduction of GSTP1 expression by DNA methylation correlates with clinicopathological features in pituitary adenomas. AB - Pi-class glutathione-S-transferase (GSTP1) located on chromosome 11q13 encodes a phase II metabolic enzyme that detoxifies reactive electrophilic intermediates. GSTP1 plays an important role in the protecting cells from cytotoxic and carcinogenic agents and is expressed in normal tissues at variable levels in different cell types. Altered GSTP1 activity and expression have been reported in many tumors and this is largely due to GSTP1 DNA hypermethylation. The role of GSTP1 in pituitary tumorigenesis has not been investigated. In this study, we evaluated the GSTP1 expression level and GSTP1 DNA methylation status in a series of pituitary adenomas. Using immunohistochemistry, we identified expression of GSTP1 in all of the various normal hormone-producing adenohypophysial cell types. In pituitary adenomas, loss or reduced expression of GSTP1 was detected in 27 of 53 tumors (50.9%). Expression of GSTP1 was significantly lower in invasive adenomas than in noninvasive adenomas (P<0.05). Using methylation-specific polymerase chain reaction (MS-PCR), GSTP1 DNA promoter hypermethylation was detected in adenomas (38 of 53, 71.7%) but not in normal tissues. GSTP1 methylation was more frequent in grade II, III, and IV tumors (66.7, 85, and 83%, respectively) than in grade I tumors (33%, P<0.05). In addition, the frequency of GSTP1 methylation was higher in invasive tumors (85%) than in noninvasive tumors (59%; P<0.05). Methylation status correlated with significant downregulation of GSTP1 expression; the frequency of GSTP1 methylation was higher in tumors with reduced-GSTP1 expression (85%) than in tumors with normal or high GSTP1 expression (54%; P<0.05). These data indicate that GSTP1 inactivation through CpG hypermethylation is common in pituitary adenomas and may contribute to aggressive pituitary tumor behavior. PMID- 18425081 TI - Ezrin expression is associated with hepatocellular carcinoma possibly derived from progenitor cells and early recurrence after surgical resection. AB - Heterogeneous biological characteristics of hepatocellular carcinoma may be attributed to the cellular origin of the tumor. Patients with hepatocellular carcinoma probably derived from hepatic progenitor cells had early tumor recurrence after surgical resection or liver transplantation, suggesting that these tumors have aggressive characteristics. Ezrin, a member of the ERM (ezrin radixin-moesin) cytoskeleton-associated protein family, is highly expressed in several types of human cancers and correlations between its immunoreactivity and patient outcome have been shown. In this study, ezrin expression, as well as cytokeratin19 and cytokeratin 7 expression, which are regarded as progenitor cell/ductular markers were immunohistochemically assessed in cases of hepatocellular carcinoma. In normal livers, ezrin expression was not found in any cell types, whereas cytokeratin 7 and cytokeratin 19 were exclusively stained in bile duct cells. In contrast, in livers with chronic hepatitis or cirrhosis, positive ezrin expression was observed in ductular reactions with strong intensity and intermediate hepatobiliary cells with various intensity. Of 77 cases of hepatocellular carcinoma, 28 (36%) had positive ezrin expression, 32 (42%) had cytokeratin 7 expression, and 11 (14%) had cytokeratin 19 expression. Ezrin expression in hepatocellular carcinoma was significantly associated with cytokeratin 19 expression, but not with cytokeratin 7 expression. Patients with ezrin-positive hepatocellular carcinoma had a significantly higher prevalence of elevated serum alpha-fetoprotein. Patients with immunohistochemical ezrin positive hepatocellular carcinoma demonstrated significantly shorter recurrence free and overall survival compared to patients with ezrin-negative hepatocellular carcinoma. Multivariate analysis revealed positive ezrin expression and multiple tumors to be independently associated with early recurrence in patients with hepatocellular carcinoma after curative surgical resection. These results suggested that hepatocellular carcinoma with ezrin expression might be at least partly derived from hepatic progenitor cells. Measurement of ezrin expression might be used to identify patients with an increased risk of early recurrence. PMID- 18425082 TI - Histological heterogeneity and somatic mtDNA mutations in gastric intraepithelial neoplasia. AB - Somatic mutations of mitochondrial DNA (mtDNA) are associated with various types of human cancer. To elucidate their role in gastric carcinogenesis, we analyzed mutations in the displacement loop region of mtDNA in 24 paraffin-embedded gastric intraepithelial neoplasias (formerly dysplasia) from a high gastric cancer risk area in northern Italy. Helicobacter pylori infection was assessed by histological examination (Giemsa staining). Gastritis was classified according to the guidelines of the Updated Sydney System. The mtDNA displacement loop region was amplified and sequenced from gastric intraepithelial neoplasia samples and adjacent non-neoplastic gastric mucosa. The gastric intraepithelial neoplasias were divided into two groups by their association with H. pylori gastritis. Group A with lesions arising on a background of H. pylori-positive gastritis contained 7 patients, and group B with lesions associated with H. pylori-negative gastritis contained 17 patients. Group A had a larger proportion of high-grade lesions than group B and showed a foveolar phenotype (type II dysplasia). Group B had a larger proportion of cases with mtDNA displacement loop region mutations than group A (P=0.004, Fisher's exact test) and exhibited an intestinal phenotype. No evidence of heteroplasmic variants in the mtDNA displacement loop, suggestive of mutations, was detected in gastric biopsies from 25 H. pylori-negative subjects and 60 cancer-unaffected H. pylori-positive patients. These results provide further evidence for the morphologic and mtDNA biomolecular differences of gastric intraepithelial neoplasias, and suggest the existence of two distinct pathways to gastric cancer--corpus-dominant H. pylori gastritis and the atrophy metaplasia pathway. PMID- 18425084 TI - Therapeutic targeting: a crucible for individualized medicine. PMID- 18425088 TI - Are trials of intravascular infusions of autologous mesenchymal stem cells in patients with multiple system atrophy currently justified, and are they effective? AB - Scientists and patients are keen to apply new experimental techniques to human diseases, especially currently untreatable progressive neurodegenerative diseases, of which multiple system atrophy (MSA) is an example. We argue that it behooves us to first have robust preclinical experimental evidence before launching into such studies in humans, and to control as much as possible for confounding effects when such trials come to be performed. PMID- 18425089 TI - Actin in action: the interplay between the actin cytoskeleton and synaptic efficacy. AB - Synapse regulation exploits the capacity of actin to function as a stable structural component or as a dynamic filament. Beyond its well-appreciated role in eliciting visible morphological changes at the synapse, the emerging picture points to an active contribution of actin to the modulation of the efficacy of pre- and postsynaptic terminals. Moreover, by engaging distinct pools of actin and divergent signalling pathways, actin-dependent morphological plasticity could be uncoupled from modulation of synaptic strength. The aim of this Review is to highlight some of the recent progress in elucidating the role of the actin cytoskeleton in synaptic function. PMID- 18425090 TI - Plasticity of intrinsic neuronal properties in CNS disorders. AB - The input-output relationship of neuronal networks depends both on their synaptic connectivity and on the intrinsic properties of their neuronal elements. In addition to altered synaptic properties, profound changes in intrinsic neuronal properties are observed in many CNS disorders. These changes reflect alterations in the functional properties of dendritic and somatic voltage- and Ca2+-gated ion channels. The molecular mechanisms underlying this intrinsic plasticity comprise the highly specific transcriptional or post-transcriptional regulation of ion channel expression, trafficking and function. The studies reviewed here show that intrinsic plasticity, in conjunction with synaptic plasticity, can fundamentally alter the input-output properties of neuronal networks in CNS disorders. PMID- 18425091 TI - General anaesthesia: from molecular targets to neuronal pathways of sleep and arousal. AB - The mechanisms through which general anaesthetics, an extremely diverse group of drugs, cause reversible loss of consciousness have been a long-standing mystery. Gradually, a relatively small number of important molecular targets have emerged, and how these drugs act at the molecular level is becoming clearer. Finding the link between these molecular studies and anaesthetic-induced loss of consciousness presents an enormous challenge, but comparisons with the features of natural sleep are helping us to understand how these drugs work and the neuronal pathways that they affect. Recent work suggests that the thalamus and the neuronal networks that regulate its activity are the key to understanding how anaesthetics cause loss of consciousness. PMID- 18425093 TI - Pushing the limits. PMID- 18425092 TI - Choices in neuroscience careers. AB - How do I choose a mentor? How do I decide what field of neuroscience to work in? Should I consider doing research in industry? Most students and postdoctoral researchers aiming for a successful career in neuroscience ask themselves these questions. In this article, Nature Reviews Neuroscience asks four successful neuroscientists for their thoughts on the factors one should consider when making these decisions. We hope that this Viewpoint will serve as a useful resource for junior neuroscientists who have to make important and sometimes difficult decisions that might have long-lasting consequences for their careers. PMID- 18425094 TI - Autoimmunity and transplantation: a meeting at the crossroads in Berlin. PMID- 18425095 TI - T cell heterogeneity: firmly fixed, predominantly plastic or merely malleable? PMID- 18425096 TI - Moving out: mobilizing activated T cells from lymphoid tissues. PMID- 18425097 TI - CD3varepsilon: PeRuSing for positive selection. PMID- 18425098 TI - 'Fine tuning' TLR signaling. PMID- 18425099 TI - Cross-presentation: avoiding trafficking chaos? PMID- 18425101 TI - Outside the box. PMID- 18425102 TI - Natural killer cells: diversity in search of a niche. AB - Over the years, the enigmatic lifestyle of natural killer (NK) cells has fascinated immunologists. Elucidating the principles governing NK cell biology is a constant source of surprise, as these lymphocytes often break the rules of lymphocyte 'behavior' established by their T and B cell counterparts. In this Focus issue, experts address how NK cells have secured a fundamental place in the immune system, how they 'see' their targets and how they participate in and influence immune responses. We now need to understand how environmental 'niches' condition the generation and function of distinct NK cell subsets in vivo. Ultimately, we can hope that unraveling the mysteries of how these atypical lymphocytes function will have an impact on human disease therapies. PMID- 18425103 TI - Natural killer cell recognition of missing self. PMID- 18425104 TI - Mistaken notions about natural killer cells. AB - As natural killer (NK) cells were first described more than 30 years ago--a lifetime in recent immunological history--this is a good time to reflect on their transition from outcasts of mainstream immunology to prominent players in innate immunity. Whereas much of our existing knowledge about NK cells is taken as longstanding fact, it may be surprising to immunologists of a younger vintage, particularly those working on NK cells today, that some of this knowledge was, at least initially, unexpected. In this Perspective, I take an unconventional approach to discussing our progress in understanding NK cells. PMID- 18425105 TI - Natural killer cell-directed therapies: moving from unexpected results to successful strategies. AB - Natural killer (NK) cells influence innate and adaptive immune host defenses. Existing data indicate that manipulating the balance between inhibitory and activating NK receptor signals, the sensitivity of target cells to NK cell mediated apoptosis, and NK cell cross-talk with dendritic cells might hold therapeutic promise. Efforts to modulate NK cell trafficking into inflamed tissues and/or lymph nodes, and to counteract NK cell suppressors, might also prove fruitful in the clinic. However, deeper investigation into the benefits of combination therapy, greater understanding of the functional distinctions between NK cell subsets, and design of new tools to monitor NK cell activity are needed to strengthen our ability to harness the power of NK cells for therapeutic aims. PMID- 18425106 TI - Up on the tightrope: natural killer cell activation and inhibition. AB - Natural killer (NK) cells circulate through the blood, lymphatics and tissues, on patrol for the presence of transformed or pathogen-infected cells. As almost all NK cell receptors bind to host-encoded ligands, signals are constantly being transmitted into NK cells, whether they interact with normal or abnormal cells. The sophisticated repertoire of activating and inhibitory receptors that has evolved to regulate NK cell activity ensures that NK cells protect hosts against pathogens, yet prevents deleterious NK cell-driven autoimmune responses. Here I highlight recent advances in our understanding of the structural properties and signaling pathways of the inhibitory and activating NK cell receptors, with a particular focus on the ITAM-dependent activating receptors, the NKG2D-DAP10 receptor complexes and the CD244 receptor system. PMID- 18425107 TI - Functions of natural killer cells. AB - Natural killer (NK) cells are effector lymphocytes of the innate immune system that control several types of tumors and microbial infections by limiting their spread and subsequent tissue damage. Recent research highlights the fact that NK cells are also regulatory cells engaged in reciprocal interactions with dendritic cells, macrophages, T cells and endothelial cells. NK cells can thus limit or exacerbate immune responses. Although NK cells might appear to be redundant in several conditions of immune challenge in humans, NK cell manipulation seems to hold promise in efforts to improve hematopoietic and solid organ transplantation, promote antitumor immunotherapy and control inflammatory and autoimmune disorders. PMID- 18425110 TI - Human metabolic phenotype diversity and its association with diet and blood pressure. AB - Metabolic phenotypes are the products of interactions among a variety of factors dietary, other lifestyle/environmental, gut microbial and genetic. We use a large scale exploratory analytical approach to investigate metabolic phenotype variation across and within four human populations, based on 1H NMR spectroscopy. Metabolites discriminating across populations are then linked to data for individuals on blood pressure, a major risk factor for coronary heart disease and stroke (leading causes of mortality worldwide). We analyse spectra from two 24 hour urine specimens for each of 4,630 participants from the INTERMAP epidemiological study, involving 17 population samples aged 40-59 in China, Japan, UK and USA. We show that urinary metabolite excretion patterns for East Asian and western population samples, with contrasting diets, diet-related major risk factors, and coronary heart disease/stroke rates, are significantly differentiated (P < 10(-16)), as are Chinese/Japanese metabolic phenotypes, and subgroups with differences in dietary vegetable/animal protein and blood pressure. Among discriminatory metabolites, we quantify four and show association (P < 0.05 to P < 0.0001) of mean 24-hour urinary formate excretion with blood pressure in multiple regression analyses for individuals. Mean 24-hour urinary excretion of alanine (direct) and hippurate (inverse), reflecting diet and gut microbial activities, are also associated with blood pressure of individuals. Metabolic phenotyping applied to high-quality epidemiological data offers the potential to develop an area of aetiopathogenetic knowledge involving discovery of novel biomarkers related to cardiovascular disease risk. PMID- 18425111 TI - Evolution of metal hyperaccumulation required cis-regulatory changes and triplication of HMA4. AB - Little is known about the types of mutations underlying the evolution of species specific traits. The metal hyperaccumulator Arabidopsis halleri has the rare ability to colonize heavy-metal-polluted soils, and, as an extremophile sister species of Arabidopsis thaliana, it is a powerful model for research on adaptation. A. halleri naturally accumulates and tolerates leaf concentrations as high as 2.2% zinc and 0.28% cadmium in dry biomass. On the basis of transcriptomics studies, metal hyperaccumulation in A. halleri has been associated with more than 30 candidate genes that are expressed at higher levels in A. halleri than in A. thaliana. Some of these genes have been genetically mapped to broad chromosomal segments of between 4 and 24 cM co-segregating with Zn and Cd hypertolerance. However, the in planta loss-of-function approaches required to demonstrate the contribution of a given candidate gene to metal hyperaccumulation or hypertolerance have not been pursued to date. Using RNA interference to downregulate HMA4 (HEAVY METAL ATPASE 4) expression, we show here that Zn hyperaccumulation and full hypertolerance to Cd and Zn in A. halleri depend on the metal pump HMA4. Contrary to a postulated global trans regulatory factor governing high expression of numerous metal hyperaccumulation genes, we demonstrate that enhanced expression of HMA4 in A. halleri is attributable to a combination of modified cis-regulatory sequences and copy number expansion, in comparison to A. thaliana. Transfer of an A. halleri HMA4 gene to A. thaliana recapitulates Zn partitioning into xylem vessels and the constitutive transcriptional upregulation of Zn deficiency response genes characteristic of Zn hyperaccumulators. Our results demonstrate the importance of cis-regulatory mutations and gene copy number expansion in the evolution of a complex naturally selected extreme trait. The elucidation of a natural strategy for metal hyperaccumulation enables the rational design of technologies for the clean-up of metal-contaminated soils and for bio-fortification. PMID- 18425112 TI - A global assembly line for cyanobactins. AB - More than 100 cyclic peptides harboring heterocyclized residues are known from marine ascidians, sponges and different genera of cyanobacteria. Here, we report an assembly line responsible for the biosynthesis of these diverse peptides, now called cyanobactins, both in symbiotic and free-living cyanobacteria. By comparing five new cyanobactin biosynthetic clusters, we produced the prenylated antitumor preclinical candidate trunkamide in Escherichia coli culture using genetic engineering. PMID- 18425113 TI - Root growth in Arabidopsis requires gibberellin/DELLA signalling in the endodermis. AB - Gibberellins (GAs) are key regulators of plant growth and development. They promote growth by targeting the degradation of DELLA repressor proteins; however, their site of action at the cellular, tissue or organ levels remains unknown. To map the site of GA action in regulating root growth, we expressed gai, a non degradable, mutant DELLA protein, in selected root tissues. Root growth was retarded specifically when gai was expressed in endodermal cells. Our results demonstrate that the endodermis represents the primary GA-responsive tissue regulating organ growth and that endodermal cell expansion is rate-limiting for elongation of other tissues and therefore of the root as a whole. PMID- 18425114 TI - Intercellular transfer of the oncogenic receptor EGFRvIII by microvesicles derived from tumour cells. AB - Aggressive human brain tumours (gliomas) often express a truncated and oncogenic form of the epidermal growth factor receptor, known as EGFRvIII. Within each tumour only a small percentage of glioma cells may actually express EGFRvIII; however, most of the cells exhibit a transformed phenotype. Here we show that EGFRvIII can be 'shared' between glioma cells by intercellular transfer of membrane-derived microvesicles ('oncosomes'). EGFRvIII expression in indolent glioma cells stimulates formation of lipid-raft related microvesicles containing EGFRvIII. Microvesicles containing this receptor are then released to cellular surroundings and blood of tumour-bearing mice, and can merge with the plasma membranes of cancer cells lacking EGFRvIII. This event leads to the transfer of oncogenic activity, including activation of transforming signalling pathways (MAPK and Akt), changes in expression of EGFRvIII-regulated genes (VEGF, Bcl x(L), p27), morphological transformation and increase in anchorage-independent growth capacity. Thus, membrane microvesicles of cancer cells can contribute to a horizontal propagation of oncogenes and their associated transforming phenotype among subsets of cancer cells. PMID- 18425115 TI - Myeloid and lymphoid contribution to non-haematopoietic lineages through irradiation-induced heterotypic cell fusion. AB - Recent studies have suggested that regeneration of non-haematopoietic cell lineages can occur through heterotypic cell fusion with haematopoietic cells of the myeloid lineage. Here we show that lymphocytes also form heterotypic-fusion hybrids with cardiomyocytes, skeletal muscle, hepatocytes and Purkinje neurons. However, through lineage fate-mapping we demonstrate that such in vivo fusion of lymphoid and myeloid blood cells does not occur to an appreciable extent in steady-state adult tissues or during normal development. Rather, fusion of blood cells with different non-haematopoietic cell types is induced by organ-specific injuries or whole-body irradiation, which has been used in previous studies to condition recipients of bone marrow transplants. Our findings demonstrate that blood cells of the lymphoid and myeloid lineages contribute to various non haematopoietic tissues by forming rare fusion hybrids, but almost exclusively in response to injuries or inflammation. PMID- 18425117 TI - A crucial role of a high mobility group protein HMGA2 in cardiogenesis. AB - The high mobility group (HMG) of nuclear proteins regulates expression of many genes through architectural remodelling of the chromatin structure, and formation of multiprotein complexes on promoter/enhancer regions. This leads to the active transcription of their target genes. Here we show that HMGA2, a member of the HMGA sub-family of HMG proteins, has a critical function in cardiogenesis. Overexpression of HMGA2 enhanced, whereas siRNA-mediated knockdown of HMGA2 blocked, cardiomyocyte differentiation of the embryonal carcinoma cell line P19CL6. Moreover, overexpression of a dominant-negative HMGA2 or morpholino mediated knockdown of HMGA2 expression blocked normal heart formation in Xenopus laevis embryos, suggesting that HMGA2 has an important role in cardiogenesis both in vitro and in vivo. Mechanistically, HMGA2 associated with Smad1/4 and showed synergistic trans-activation of the gene for a cardiac transcription factor Nkx2.5; a conserved HMGA2 binding site was required for the promoter activity of Nkx2.5 gene, both in P19CL6 cells and in transgenic Xenopus embryos. Thus, HMGA2 is a positive regulator of Nkx2.5 gene expression and is essential for normal cardiac development. PMID- 18425116 TI - Extensive fusion of haematopoietic cells with Purkinje neurons in response to chronic inflammation. AB - Transplanted bone marrow-derived cells (BMDCs) have been reported to fuse with cells of diverse tissues, but the extremely low frequency of fusion has led to the view that such events are biologically insignificant. Nonetheless, in mice with a lethal recessive liver disease (tyrosinaemia), transplantation of wild type BMDCs restored liver function by cell fusion and prevented death, indicating that cell fusion can have beneficial effects. Here we report that chronic inflammation resulting from severe dermatitis or autoimmune encephalitis leads to robust fusion of BMDCs with Purkinje neurons and formation of hundreds of binucleate heterokaryons per cerebellum, a 10-100-fold higher frequency than previously reported. Single haematopoietic stem-cell transplants showed that the fusogenic cell is from the haematopoietic lineage and parabiosis experiments revealed that fusion can occur without irradiation. Transplantation of rat bone marrow into mice led to activation of dormant rat Purkinje neuron-specific genes in BMDC nuclei after fusion with mouse Purkinje neurons, consistent with nuclear reprogramming. The precise neurological role of these heterokaryons awaits elucidation, but their frequency in brain after inflammation is clearly much higher than previously appreciated. PMID- 18425119 TI - Angiopoietins assemble distinct Tie2 signalling complexes in endothelial cell cell and cell-matrix contacts. AB - The receptor tyrosine kinase Tie2, and its activating ligand Angiopoietin-1 (Ang1), are required for vascular remodelling and vessel integrity, whereas Ang2 may counteract these functions. However, it is not known how Tie2 transduces these different signals. Here, we show that Ang1 induces unique Tie2 complexes in mobile and confluent endothelial cells. Matrix-bound Ang1 induced cell adhesion, motility and Tie2 activation in cell-matrix contacts that became translocated to the trailing edge in migrating endothelial cells. In contrast, in contacting cells Ang1 induced Tie2 translocation to cell-cell contacts and the formation of homotypic Tie2-Tie2 trans-associated complexes that included the vascular endothelial phosphotyrosine phosphatase, leading to inhibition of paracellular permeability. Distinct signalling proteins were preferentially activated by Tie2 in the cell-matrix and cell-cell contacts, where Ang2 inhibited Ang1-induced Tie2 activation. This novel type of cellular microenvironment-dependent receptor tyrosine kinase activation may explain some of the effects of angiopoietins in angiogenesis and vessel stabilization. PMID- 18425118 TI - A pathway for phagosome maturation during engulfment of apoptotic cells. AB - Removal of apoptotic cells is critical for the physiological well-being of the organism and defects in corpse removal have been linked to disease states. Genes regulating corpse recognition and internalization have been identified, but few molecules involved in the processing of internalized corpses are known. Through a combination of targeted and unbiased reverse genetic screens in Caenorhabditis elegans, and studies in mammalian cells, we have identified genes required for maturation of apoptotic-cell-containing phagosomes. We have further ordered these candidates, which include the GTPases RAB-5 and RAB-7 and the HOPS complex, into a coherent linear pathway for the maturation of apoptotic cells within phagosomes. In depth analysis of two additional candidate genes, the phosphatidylinositol 3 kinase (PI(3)K) vps-34 (A001762) and dyn-1/dynamin, showed an accumulation of internalized, but undegraded, corpses within abnormal Rab5 negative phagosomes. We ordered these candidates in our pathway, with DYN-1 functioning upstream of VPS-34 in the recruitment and/or retention of RAB-5 to the phagosome. Finally, we have also identified a previously undescribed biochemical complex containing Vps34, dynamin and Rab5(GDP), thus providing a mechanism for Rab5 recruitment to the nascent phagosome. PMID- 18425120 TI - Differential function of Tie2 at cell-cell contacts and cell-substratum contacts regulated by angiopoietin-1. AB - Tie2 belongs to the receptor tyrosine kinase family and functions as a receptor for Angiopoietin-1 (Ang1). Gene-targeting analyses of either Ang1 or Tie2 in mice reveal a critical role of Ang1-Tie2 signalling in developmental vascular formation. It remains elusive how the Tie2 signalling pathway plays distinct roles in both vascular quiescence and angiogenesis. We demonstrate here that Ang1 bridges Tie2 at cell-cell contacts, resulting in trans-association of Tie2 in the presence of cell-cell contacts. In clear contrast, in isolated cells, extracellular matrix-bound Ang1 locates Tie2 at cell-substratum contacts. Furthermore, Tie2 activated at cell-cell or cell-substratum contacts leads to preferential activation of Akt and Erk, respectively. Microarray analyses and real-time PCR validation clearly show the differential gene expression profile in vascular endothelial cells upon Ang1 stimulation in the presence or absence of cell-cell contacts, implying downstream signalling is dependent upon the spatial localization of Tie2. PMID- 18425121 TI - Control of the establishment of aversive memory by calcineurin and Zif268. AB - Emotional memory is a rapidly acquired and persistent form of memory, and its robustness is in part determined by the initial strength of the memory. Here, we provide new evidence that the protein phosphatase calcineurin (CaN), a potent negative regulator of neuronal signaling that is known to constrain learning and memory, critically regulates the establishment of emotional memory through mechanisms involving the immediate early gene Zif268 (also known as Egr1). We found that CaN is inhibited in the amygdala during the establishment of aversive memory, but Zif268 is activated. Using inducible transgenesis in mice, we further saw that CaN inhibition and Zif268 overexpression during memory establishment strengthen the memory trace and enhance its resistance to extinction. We found that CaN inhibition correlates with increased Zif268 expression and that a common pool of proteins is regulated in the amygdala after CaN inhibition and Zif268 overexpression. Together, these findings reveal a previously unknown mechanism for the control of emotional memory that depends on CaN and Zif268. PMID- 18425122 TI - Quantal noise from human red cone pigment. AB - The rod pigment, rhodopsin, shows spontaneous isomerization activity. This quantal noise produces a dark light of approximately 0.01 photons s(-1) rod(-1) in human, setting the threshold for rod vision. The spontaneous isomerization activity of human cone pigments has long remained a mystery because the effect of a single isomerized pigment molecule in cones, unlike that in rods, is small and beyond measurement. We have now overcome this problem by expressing human red cone pigment transgenically in mouse rods in order to exploit their large single photon response, especially after genetic removal of a key negative-feedback regulation. Extrapolating the measured quantal noise of transgenic cone pigment to native human red cones, we obtained a dark rate of approximately 10 false events s(-1) cone(-1), almost 10(3)-fold lower than the overall dark transduction noise previously reported in primate cones. Our measurements provide a rationale for why mammalian red, green and blue cones have comparable sensitivities, unlike their amphibian counterparts. PMID- 18425123 TI - Cortical activity patterns predict speech discrimination ability. AB - Neural activity in the cerebral cortex can explain many aspects of sensory perception. Extensive psychophysical and neurophysiological studies of visual motion and vibrotactile processing show that the firing rate of cortical neurons averaged across 50-500 ms is well correlated with discrimination ability. In this study, we tested the hypothesis that primary auditory cortex (A1) neurons use temporal precision on the order of 1-10 ms to represent speech sounds shifted into the rat hearing range. Neural discrimination was highly correlated with behavioral performance on 11 consonant-discrimination tasks when spike timing was preserved and was not correlated when spike timing was eliminated. This result suggests that spike timing contributes to the auditory cortex representation of consonant sounds. PMID- 18425124 TI - Theta phase-specific codes for two-dimensional position, trajectory and heading in the hippocampus. AB - Temporal coding is a means of representing information by the time, as opposed to the rate, at which neurons fire. Evidence of temporal coding in the hippocampus comes from place cells, whose spike times relative to theta oscillations reflect a rat's position while running along stereotyped trajectories. This arises from the backwards shift in cell firing relative to local theta oscillations (phase precession). Here we demonstrate phase precession during place-field crossings in an open-field foraging task. This produced spike sequences in each theta cycle that disambiguate the rat's trajectory through two-dimensional space and can be used to predict movement direction. Furthermore, position and movement direction were maximally predicted from firing in the early and late portions of the theta cycle, respectively. This represents the first direct evidence of a combined representation of position, trajectory and heading in the hippocampus, organized on a fine temporal scale by theta oscillations. PMID- 18425125 TI - Single-round infectious particles enhance immunogenicity of a DNA vaccine against West Nile virus. AB - DNA vaccines encoding replication-defective viruses are safer than inactivated or live attenuated viruses but may fail to stimulate an immune response sufficient for effective vaccination. We augment the protective capacity of a capsid-deleted flavivirus DNA vaccine by co-expressing the capsid protein from a separate promoter. In transfected cells, the capsid-deleted RNA transcript is replicated and translated to produce secreted virus-like particles lacking the nucleocapsid. This RNA is also packaged with the help of co-expressed capsid protein to form secreted single-round infectious particles (SRIPs) that deliver the RNA into neighboring cells. In SRIP-infected cells, the RNA is replicated again and produces additional virus-like particles, but in the absence of capsid RNA no SRIPs are formed and no further spread occurs. Compared with an otherwise identical construct that does not encode capsid, our vaccine offers better protection to mice after lethal West Nile virus infection. It also elicits virus neutralizing antibodies in horses. This approach may enable vaccination against pathogenic flaviviruses other than West Nile virus. PMID- 18425126 TI - SmcHD1, containing a structural-maintenance-of-chromosomes hinge domain, has a critical role in X inactivation. AB - X-chromosome inactivation is the mammalian dosage compensation mechanism by which transcription of X-linked genes is equalized between females and males. In an N ethyl-N-nitrosourea (ENU) mutagenesis screen on mice for modifiers of epigenetic reprogramming, we identified the MommeD1 (modifier of murine metastable epialleles) mutation as a semidominant suppressor of variegation. MommeD1 shows homozygous female-specific mid-gestation lethality and hypomethylation of the X linked gene Hprt1, suggestive of a defect in X inactivation. Here we report that the causative point mutation lies in a previously uncharacterized gene, Smchd1 (structural maintenance of chromosomes hinge domain containing 1). We find that SmcHD1 is not required for correct Xist expression, but localizes to the inactive X and has a role in the maintenance of X inactivation and the hypermethylation of CpG islands associated with the inactive X. This finding links a group of proteins normally associated with structural aspects of chromosome biology with epigenetic gene silencing. PMID- 18425127 TI - Interpreting principal component analyses of spatial population genetic variation. AB - Nearly 30 years ago, Cavalli-Sforza et al. pioneered the use of principal component analysis (PCA) in population genetics and used PCA to produce maps summarizing human genetic variation across continental regions. They interpreted gradient and wave patterns in these maps as signatures of specific migration events. These interpretations have been controversial, but influential, and the use of PCA has become widespread in analysis of population genetics data. However, the behavior of PCA for genetic data showing continuous spatial variation, such as might exist within human continental groups, has been less well characterized. Here, we find that gradients and waves observed in Cavalli Sforza et al.'s maps resemble sinusoidal mathematical artifacts that arise generally when PCA is applied to spatial data, implying that the patterns do not necessarily reflect specific migration events. Our findings aid interpretation of PCA results and suggest how PCA can help correct for continuous population structure in association studies. PMID- 18425128 TI - A structural-maintenance-of-chromosomes hinge domain-containing protein is required for RNA-directed DNA methylation. AB - RNA-directed DNA methylation (RdDM) is a process in which dicer-generated small RNAs guide de novo cytosine methylation at the homologous DNA region. To identify components of the RdDM machinery important for Arabidopsis thaliana development, we targeted an enhancer active in meristems for methylation, which resulted in silencing of a downstream GFP reporter gene. This silencing system also features secondary siRNAs, which trigger methylation that spreads beyond the targeted enhancer region. A screen for mutants defective in meristem silencing and enhancer methylation retrieved six dms complementation groups, which included the known factors DRD1 (ref. 3; a SNF2-like chromatin-remodeling protein) and Pol IVb subunits. Additionally, we identified a previously unknown gene DMS3 (At3g49250), encoding a protein similar to the hinge-domain region of structural maintenance of chromosomes (SMC) proteins. This finding implicates a putative chromosome architectural protein that can potentially link nucleic acids in facilitating an RNAi-mediated epigenetic modification involving secondary siRNAs and spreading of DNA methylation. PMID- 18425129 TI - Mast cell activators: a new class of highly effective vaccine adjuvants. AB - Mast cells (MCs) have recently received recognition as prominent effectors in the regulation of immune cell migration to draining lymph nodes and lymphocyte activation. However, their role in the development of humoral immune responses is not clear. Here, we demonstrate that subcutaneous or nasal administration of small-molecule MC activators with vaccine antigens evokes large increases in antigen-specific serum immunoglobulin G (IgG) responses. These responses were MC dependent and correlated with increased dendritic cell and lymphocyte recruitment to draining lymph nodes. Nasal instillation of these formulations also evoked antigen-specific secretory IgA and provided protection against anthrax lethal toxin challenge in vitro and against vaccinia virus infection in vivo. Collectively, these results define the MC as an integral sensory arm of the adaptive immune system. Moreover, they highlight MC activators as a new class of vaccine adjuvants, capable of inducing protective antigen-specific immune responses through needle-free routes of administration. PMID- 18425130 TI - A GRK5 polymorphism that inhibits beta-adrenergic receptor signaling is protective in heart failure. AB - Beta-adrenergic receptor (betaAR) blockade is a standard therapy for cardiac failure and ischemia. G protein-coupled receptor kinases (GRKs) desensitize betaARs, suggesting that genetic GRK variants might modify outcomes in these syndromes. Re-sequencing of GRK2 and GRK5 revealed a nonsynonymous polymorphism of GRK5, common in African Americans, in which leucine is substituted for glutamine at position 41. GRK5-Leu41 uncoupled isoproterenol-stimulated responses more effectively than did GRK5-Gln41 in transfected cells and transgenic mice, and, like pharmacological betaAR blockade, GRK5-Leu41 protected against experimental catecholamine-induced cardiomyopathy. Human association studies showed a pharmacogenomic interaction between GRK5-Leu41 and beta-blocker treatment, in which the presence of the GRK5-Leu41 polymorphism was associated with decreased mortality in African Americans with heart failure or cardiac ischemia. In 375 prospectively followed African-American subjects with heart failure, GRK5-Leu41 protected against death or cardiac transplantation. Enhanced betaAR desensitization of excessive catecholamine signaling by GRK5-Leu41 provides a 'genetic beta-blockade' that improves survival in African Americans with heart failure, suggesting a reason for conflicting results of beta-blocker clinical trials in this population. PMID- 18425131 TI - Drug-induced cure drives conversion to a stable and protective CD8+ T central memory response in chronic Chagas disease. AB - In this study, we document the development of stable, antigen-independent CD8+ T cell memory after drug-induced cure of a chronic infection. By establishing a system for drug cure of chronic Trypanosoma cruzi infection, we present the first extensively documented case of total parasite clearance after drug treatment of this infection. Cure resulted in the emergence of a stable, parasite-specific CD8+ T cell population with the characteristics of central memory cells, based upon expression of CD62L, CCR7, CD127, CD122, Bcl-2 and a reduced immediate in vivo CTL function. CD8+ T cells from treated and cured mice also expanded more rapidly and provided greater protection following challenge than those from chronically infected mice. These results show that complete pathogen clearance results in stable, antigen-independent and protective T cell memory, despite the potentially exhausting effects of prior long-term exposure to antigen in this chronic infection. PMID- 18425132 TI - Restoration of lymphoid organ integrity through the interaction of lymphoid tissue-inducer cells with stroma of the T cell zone. AB - The generation of lymphoid microenvironments in early life depends on the interaction of lymphoid tissue-inducer cells with stromal lymphoid tissue organizer cells. Whether this cellular interface stays operational in adult secondary lymphoid organs has remained elusive. We show here that during acute infection with lymphocytic choriomeningitis virus, antiviral cytotoxic T cells destroyed infected T cell zone stromal cells, which led to profound disruption of secondary lymphoid organ integrity. Furthermore, the ability of the host to respond to secondary antigens was lost. Restoration of the lymphoid microanatomy was dependent on the proliferative accumulation of lymphoid tissue-inducer cells in secondary lymphoid organs during the acute phase of infection and lymphotoxin alpha(1)beta(2) signaling. Thus, crosstalk between lymphoid tissue-inducer cells and stromal cells is reactivated in adults to maintain secondary lymphoid organ integrity and thereby contributes to the preservation of immunocompetence. PMID- 18425133 TI - High-frequency micromechanical resonators from aluminium-carbon nanotube nanolaminates. AB - At micro- and nanoscales, materials with high Young's moduli and low densities are of great interest for high-frequency micromechanical resonator devices. Incorporating carbon nanotubes (CNTs), with their unmatched properties, has added functionality to many man-made composites. We report on the fabrication of < or = 100-nm-thick laminates by sputter-deposition of aluminium onto a two-dimensional single-walled CNT network. These nanolaminates--composed of Al, its native oxide Al(2)O(3) and CNTs--are fashioned, in a scalable manner, into suspended doubly clamped micromechanical beams. Dynamic flexural measurements show marked increases in resonant frequencies for nanolaminates with Al-CNT laminae. Such increases, further supported by quasi-static flexural measurements, are partly attributable to enhancements in elastic properties arising from the addition of CNTs. As a consequence, these nanolaminate micromechanical resonators show significant suppression of mechanical nonlinearity and enhanced strength, both of which are advantageous for practical applications and analogous to biological nanocomposites, similarly composed of high-aspect-ratio, mechanically superior mineral platelets in a soft protein matrix. PMID- 18425134 TI - Bulk superconductivity at 38 K in a molecular system. AB - C(60)-based solids are archetypal molecular superconductors with transition temperatures (Tc) as high as 33 K (refs 2-4). Tc of face-centred-cubic (f.c.c.) A(3)C(60) (A=alkali metal) increases monotonically with inter C(60) separation, which is controlled by the A(+) cation size. As Cs(+) is the largest such ion, Cs(3)C(60) is a key material in this family. Previous studies revealing trace superconductivity in Cs(x)C(60) materials have not identified the structure or composition of the superconducting phase owing to extremely small shielding fractions and low crystallinity. Here, we show that superconducting Cs(3)C(60) can be reproducibly isolated by solvent-controlled synthesis and has the highest Tc of any molecular material at 38 K. In contrast to other A(3)C(60) materials, two distinct cubic Cs(3)C(60) structures are accessible. Although f.c.c. Cs(3)C(60) can be synthesized, the superconducting phase has the A15 structure based uniquely among fullerides on body-centred-cubic packing. Application of hydrostatic pressure controllably tunes A15 Cs(3)C(60) from insulating at ambient pressure to superconducting without crystal structure change and reveals a broad maximum in Tc at approximately 7 kbar. We attribute the observed Tc maximum as a function of inter C(60)separation--unprecedented in fullerides but reminiscent of the atom-based cuprate superconductors--to the role of strong electronic correlations near the metal-insulator transition onset. PMID- 18425135 TI - Probing the structure of heterogeneous diluted materials by diffraction tomography. AB - The advent of nanosciences calls for the development of local structural probes, in particular to characterize ill-ordered or heterogeneous materials. Furthermore, because materials properties are often related to their heterogeneity and the hierarchical arrangement of their structure, different structural probes covering a wide range of scales are required. X-ray diffraction is one of the prime structural methods but suffers from a relatively poor detection limit, whereas transmission electron analysis involves destructive sample preparation. Here we show the potential of coupling pencil-beam tomography with X-ray diffraction to examine unidentified phases in nanomaterials and polycrystalline materials. The demonstration is carried out on a high-pressure pellet containing several carbon phases and on a heterogeneous powder containing chalcedony and iron pigments. The present method enables a non-invasive structural refinement with a weight sensitivity of one part per thousand. It enables the extraction of the scattering patterns of amorphous and crystalline compounds with similar atomic densities and compositions. Furthermore, such a diffraction-tomography experiment can be carried out simultaneously with X-ray fluorescence, Compton and absorption tomographies, enabling a multimodal analysis of prime importance in materials science, chemistry, geology, environmental science, medical science, palaeontology and cultural heritage. PMID- 18425136 TI - A sensitivity-enhanced field-effect chiral sensor. AB - Organic thin-film transistor sensors have been recently attracting the attention of the plastic electronics community for their potential exploitation in novel sensing platforms. Specificity and sensitivity are however still open issues: in this respect chiral discrimination-being a scientific and technological achievement in itself--is indeed one of the most challenging sensor bench-tests. So far, conducting-polymer solid-state chiral detection has been carried out at part-per-thousand concentration levels. Here, a novel chiral bilayer organic thin film transistor gas sensor--comprising an outermost layer with built-in enantioselective properties-is demonstrated to show field-effect amplified sensitivity that enables differential detection of optical isomers in the tens-of parts-per-million concentration range. The ad-hoc-designed organic semiconductor endowed with chiral side groups, the bilayer structure and the thin-film transistor transducer provide a significant step forward in the development of a high-performance and versatile sensing platform compatible with flexible organic electronic technologies. PMID- 18425137 TI - Fluorescent protein FRET pairs for ratiometric imaging of dual biosensors. AB - Fluorescence resonance energy transfer (FRET) with fluorescent proteins is a powerful method for detection of protein-protein interactions, enzyme activities and small molecules in the intracellular milieu. Aided by a new violet-excitable yellow-fluorescing variant of Aequorea victoria GFP, we developed dual FRET-based caspase-3 biosensors. Owing to their distinct excitation profiles, each FRET biosensor can be ratiometrically imaged in the presence of the other. PMID- 18425138 TI - Monovalent, reduced-size quantum dots for imaging receptors on living cells. AB - We describe a method to generate monovalent quantum dots (QDs) using agarose gel electrophoresis. We passivated QDs with a carboxy-terminated polyethylene-glycol ligand, yielding particles with half the diameter of commercial QDs, which we conjugated to a single copy of a high-affinity targeting moiety (monovalent streptavidin or antibody to carcinoembryonic antigen) to label cell-surface proteins. The small size improved access of QD-labeled glutamate receptors to neuronal synapses, and monovalency prevented EphA3 tyrosine kinase activation. PMID- 18425140 TI - Straightforward ladder sequencing of peptides using a Lys-N metalloendopeptidase. AB - We introduce a method for sequencing peptides by mass spectrometry using a metalloendopeptidase that cleaves proteins at the amino side of lysine (Lys-N). When analyzed by electron transfer dissociation (ETD)-based mass spectrometric sequencing, Lys-N-digested peptides that contain a single lysine residue produce spectra dominated by c-type fragment ions, providing simple ladders for sequence determination. This method should be a valuable strategy for de novo sequencing and the analysis of post-translational modifications. PMID- 18425139 TI - Modeling lymphangiogenesis in a three-dimensional culture system. AB - A lack of appropriate in vitro models of three-dimensional lymph vessel growth hampers the study of lymphangiogenesis. We developed a lymphatic ring assay--a potent, reproducible and quantifiable three-dimensional culture system for lymphatic endothelial cells that reproduces spreading of endothelial cells from a pre-existing vessel, cell proliferation, migration and differentiation into capillaries. In the assay, mouse thoracic duct fragments are embedded in a collagen gel, leading to the formation of lumen-containing lymphatic capillaries, which we assessed by electron microscopy and immunostaining. We developed a computerized method to quantify the lymphatic network. By applying this model to gene-deficient mice, we found evidence for involvement of the matrix metalloproteinase, MMP-2, in lymphangiogenesis. The lymphatic ring assay bridges the gap between two-dimensional in vitro models and in vivo models of lymphangiogenesis, can be used to exploit the potential of existing transgenic mouse models, and rapidly identify regulators of lymphangiogenesis. PMID- 18425141 TI - Aminoacylation of tRNA with phosphoserine for synthesis of cysteinyl-tRNA(Cys). AB - Cysteinyl-tRNA(Cys) (Cys-tRNA(Cys)) is required for translation and is typically synthesized by cysteinyl-tRNA synthetase (CysRS). However, Methanocaldococcus jannaschii synthesizes Cys-tRNA(Cys) by an indirect pathway, whereby O phosphoseryl-tRNA synthetase (SepRS) acylates tRNA(Cys) with phosphoserine (Sep), and Sep-tRNA-Cys-tRNA synthase (SepCysS) converts the tRNA-bound phosphoserine to cysteine. We show here that M. jannaschii SepRS differs from CysRS by recruiting the m1G37 modification as a determinant for aminoacylation, and in showing limited discrimination against mutations of conserved nucleotides. Kinetic and binding measurements show that both SepRS and SepCysS bind the reaction intermediate Sep-tRNA(Cys) tightly, and these two enzymes form a stable binary complex that promotes conversion of the intermediate to the product and sequesters the intermediate from binding to elongation factor EF-1alpha or infiltrating into the ribosome. These results highlight the importance of the protein binary complex for efficient synthesis of Cys-tRNA(Cys). PMID- 18425142 TI - Phosphorylation of human PRP28 by SRPK2 is required for integration of the U4/U6 U5 tri-snRNP into the spliceosome. AB - Several protein kinases, including SRPK1 and SRPK2, have been implicated in spliceosome assembly and catalytic activation. However, little is known about their targets. Here we show that SRPK1 is predominantly associated with U1 small nuclear ribonucleoprotein (snRNP), whereas SRPK2 associates with the U4/U6-U5 tri snRNP. RNAi-mediated depletion in HeLa cells showed that SRPK2 is essential for cell viability, and it is required for spliceosomal B complex formation. SRPK2 knock down results in hypophosphorylation of the arginine-serine (RS) domain containing human PRP28 protein (PRP28, also known as DDX23), and destabilizes PRP28 association with the tri-snRNP. Immunodepletion of PRP28 from HeLa cell nuclear extract and complementation studies revealed that PRP28 phosphorylation is required for its stable association with the tri-snRNP and for tri-snRNP integration into the B complex. Our results demonstrate a role for SRPK2 in splicing and reveal a previously unknown function for PRP28 in spliceosome assembly. PMID- 18425144 TI - What is the optimal dosage of valganciclovir as preemptive therapy for CMV infection in allogeneic hematopoietic SCT? PMID- 18425143 TI - A role for ubiquitin in the spliceosome assembly pathway. AB - The spliceosome uses numerous strategies to regulate its function in mRNA maturation. Ubiquitin regulates many cellular processes, but its potential roles during splicing are unknown. We have developed a new strategy that reveals a direct role for ubiquitin in the dynamics of splicing complexes. A ubiquitin mutant (I44A) that can enter the conjugation pathway but is compromised in downstream functions diminishes splicing activity by reducing the levels of the U4/U6-U5 small nuclear ribonucleoprotein (snRNP). Similarly, an inhibitor of ubiquitin's protein-protein interactions, ubistatin A, reduces U4/U6-U5 triple snRNP levels in vitro. When ubiquitin interactions are blocked, ATP-dependent disassembly of purified U4/U6-U5 particles is accelerated, indicating a direct role for ubiquitin in repressing U4/U6 unwinding. Finally, we show that the conserved splicing factor Prp8 is ubiquitinated within purified triple snRNPs. These results reveal a previously unknown ubiquitin-dependent mechanism for controlling the pre-mRNA splicing pathway. PMID- 18425145 TI - Contemporary analysis of the influence of acute kidney injury (AKI) after myeloablative hematopoietic cell transplantation on long-term patient's survival. PMID- 18425146 TI - Comparable engraftment and chimerism kinetics using oral and intravenous fludarabine as part of a reduced intensity conditioning regimen. PMID- 18425147 TI - Subsets of CD34+ and early engraftment kinetics in allogeneic peripheral SCT for AML. AB - This study aimed to identify which graft product subset of CD34+ cells might be the most predictive of early hematopoietic recovery following allogeneic peripheral SCT (allo-PBSCT). The relationship between the number of 'mature' subsets of CD34+ cells (CD34+/CD33+, CD34+/CD38+, CD34+/DR+ and CD34+/CD133-) and 'immature' subsets of CD34+ cells (CD34+/CD33-, CD34+/CD38-, CD34+/DR- and CD34+/CD133+) and early neutrophil and platelet engraftment were studied in a homogeneous series (for disease, pre transplant chemotherapy, conditioning regimen and GVHD prophylaxis) of 30 AML patients after allo-PBSCT from HLA identical siblings. In our experience, the total CD34+/CD133+ cell number was inversely correlated with the days required for the recovery of 0.5 x 10(9)/l neutrophils (r=or-0.82, P=0.02) and platelets of 20 x 10(9)/l (r=or-0.60, P=0.06); this correlation was better than the total CD34+ cell dose and neutrophil (r=or-0.70, P=0.04) and platelet engraftment (r=or-0.56, P=0.07). We suggest that a high number of CD34+/CD133+ PBSC may be associated with faster neutrophil and platelet recovery; these findings may help to predict the repopulating capacity of PBSC in patients after allo-PBSCT, especially when a relatively low number of CD34+ cells is infused. PMID- 18425148 TI - Medication error in New Zealand--time to act. PMID- 18425149 TI - Personalised medicine in New Zealand. PMID- 18425150 TI - Health services research: an essential part of a wider spectrum of national health information? PMID- 18425151 TI - Preventable medication-related events in hospitalised children in New Zealand. AB - AIMS: To evaluate the frequency and characteristics of preventable medication related events in hospitalised children, to determine the yield of several methods for identifying them and to recommend priorities for prevention. METHODS: A prospective observational cohort study was conducted over a 12-week period on the paediatric wards at a university-affiliated urban general hospital in New Zealand. For all admissions of greater than 24 hours, medication-related events were identified using a multifaceted approach and subsequently classified independently by three reviewers (using a standardised reviewer form) by event type, type of error, stage of the medication process, and preventability. RESULTS: There were 495 eligible study patients, who had 520 admissions and 3037 patient days of admission, during which 3160 medication orders were written. Of 761 medication-related events reported during the study period, 630 (83.3%) were identified by chart review; 111 (14.6%) by a voluntary staff quality improvement reporting system; 16 (2.1%) by interview of parents; and 4 (0.53%) events via the concurrent routine hospital-incident reporting system. Excluding duplicate reports and practice-related issues, a total of 696 study patient-specific events were included in the analysis. Excluding the inconsequential events (trivial rule violation and 'other' categories), the majority [368/399 (92.2%)] of events were found to be preventable; comprising 38/67 (56.7%) ADEs, 75/77 (97.4%) potential ADEs, and all 255 (100%) harmless medication errors. Most commonly implicated in preventable ADEs and potential ADEs were, event rate (95%CI): improper dose and the prescribing stage-35 (29 to 42) and 74 (64 to 84) respectively per 1000 patient days; and antibacterial agents and the intravenous route of administration 21 (17 to 25) and 11 (10 to 13) respectively per 100 medication orders. CONCLUSIONS: Preventable medication-related events occur commonly in the paediatric inpatient setting, and importantly over half of the events that caused patient harm were deemed preventable. Voluntary staff reporting in a quality improvement environment was found to be inferior to chart review for identifying events, but a vast improvement on the conventional incident reporting system. Most commonly implicated in the harmful or potentially harmful preventable events, and hence the best targets for prevention are dosing errors, particularly during the prescribing stage of the medication use process, and use of antibacterial agents, particularly when administered by the intravenous route. PMID- 18425152 TI - Allele frequency differences of cytochrome P450 polymorphisms in a sample of New Zealand Maori. AB - AIMS: To determine the prevalence of functional alleles for drug metabolising genes in a sample of Maori and compare allele frequencies with Caucasians estimates. PROCEDURES: DNA from 60 Maori volunteers was genotyped for cytochrome P450 polymorphisms--CYP2A6, CYP2C9, CYP2C19, and CYP2D6--and allele frequencies calculated and compared with Caucasian estimates. RESULTS: Absolute allele frequency differences between Maori and Caucasian groups ranged from 1% to 16% for the polymorphisms tested. CONCLUSIONS: Functional allele frequencies of drug metabolising genes differed between Maori and European groups warranting larger general population surveys. These findings may also bear thinking about when conducting pharmacogenetic studies or clinical trials in New Zealand cohorts because patients with Maori ancestry may respond differently to certain medicines based on genotype. PMID- 18425153 TI - Surveillance of vaccine breakthrough cases following MeNZB vaccination. AB - AIM: To describe and investigate epidemic strain serogroup B meningococcal disease in recipients of the meningococcal vaccine, MeNZB. METHOD: Epidemic strain meningococcal disease cases in vaccine recipients were identified by matching disease notification and laboratory data against the National Immunisation Register. Descriptive analyses were undertaken for disease cases aged under 20 years and vaccine breakthrough cases (epidemic strain meningococcal disease cases with onset 28 or more days after receipt of the third MeNZB dose). Questionnaires were sent to hospital clinicians requesting medical histories and laboratory test results for vaccine breakthrough cases. A committee reviewed this information to assess immune competence in these cases. RESULTS: From the start of the meningococcal B immunisation programme in July 2004 to the end of 2006, 34 vaccine breakthrough cases were identified. No underlying host factors were identified that explained disease occurrence for the 30 cases (88.2%) for whom questionnaires were completed. For 12 (35.3%) cases all requested laboratory tests to assess immune competence were performed and these subjects were judged to be immune competent. CONCLUSION: While epidemic strain meningococcal disease incidence has fallen dramatically with the introduction of the vaccine, these early results confirm the expectation that vaccine breakthrough cases will occur in immune competent individuals given the anticipated vaccine effectiveness of approximately 75%. PMID- 18425155 TI - Correlation of physician seniority with increased emergency department efficiency during a resident doctors' strike. AB - AIM: Physician seniority has increasingly been shown to correlate with improved clinical outcomes. However few studies examine the relationship between treating doctor experience and the efficiency of emergency care systems. We explored the hypothesis that increased seniority of emergency department (ED) medical staff would result in improved ED efficiency. METHOD: This was prospective observational study conducted at the ED of Waikato Hospital, a 650-bed university affiliated teaching hospital. All patient presentations during a 5-day resident doctors' strike when the ED was staffed by senior physicians, and the corresponding normally staffed days of the subsequent calendar week were examined. Patient waiting times, time seen to disposition, and total ED length of stay were recorded according to Australasian Triage Score (ATS). RESULTS: 608 and 683 patient presentations were recorded during the strike and non-strike period respectively. Waiting times were reduced for ATS3 (43.8 vs 73.6 minutes, p<0.001) and ATS4 (53.7 vs 82.0 minutes, p<0.001) during the strike period. Time seen to disposition were reduced for ATS2 (147.9 vs 255.1 minutes, p=0.001) and ATS3 (119.9 vs 165.0 minutes, p<0.001) during the strike period. ED length of stay was reduced for ATS2 (162.6 vs 278.6 minutes, p<0.001), ATS3 (161.9 vs 238.4 minutes, p<0.001), and ATS4 (134.1 vs 179.2 minutes, p<0.001) during the strike period. No difference was observed in patient walkout, ED mortality, 48-hour mortality, or 30-day unscheduled representation rates. CONCLUSIONS: Increasing seniority of front line ED staff during a period of resident doctors' strike action was associated with increased efficiency of ED patient processing. Early specialist involvement with ED patients may replicate these efficiencies during periods of normal departmental operation. PMID- 18425154 TI - Capitation funding of primary health organisations in New Zealand: are enrolled populations being funded according to need? AB - AIM: To determine whether the three main funding formulas for Primary Health Organisations achieved a stated aim of the Primary Health Care Strategy to fund enrolled populations according to need. METHODS: National data were obtained from the Ministry of Health for a 12-month period beginning in April 2004: these included demographic characteristics of the enrolled Primary Health Organisation population, plus rates tables for: First-Contact Services, Services to Improve Access, and Health Promotion. Funding for Access and Interim practices for four quarters was calculated for each of these three funding streams. Analysis of the demographic characteristics of Access and Interim practices was undertaken. RESULTS: Maori and Pacific peoples made up a greater proportion of the Access population than the Interim, had higher rates of deprivation than the non Maori/non-Pacific population, and demonstrated a younger age distribution. The first quarter (April 2004-June 2004) showed there was preferential funding for Access PHOs and in particular high-needs groups. In quarter two, this level of preferential funding had diminished, coinciding with the introduction of increased government funding for all Interim enrolees aged 65 and over. CONCLUSIONS: The greater funding for Access enrolees was notably eroded with the introduction of Access-level funding for those aged 65+ in Interim PHOs. Since these data were analysed all remaining Interim age groups have shifted to Access level funding, benefiting non-Maori /non-Pacific in Interim PHOs. The rapid shift to Access-level funding for First Contact Services has seen a continued erosion of the redistributive effect of the original needs-based formulas. A system cannot be considered equitable if some members of society are not realising their health potential, and financing of primary care should remain redistributive until such a time as this objective is attained. PMID- 18425156 TI - Evidence-based recommendations for hand hygiene for health care workers in New Zealand. AB - AIM: The aim of this systematic review is to establish evidence-based recommendations for hand hygiene for health care workers in New Zealand. METHODS: Using a systematic approach to literature searching, relevant studies were retrieved and evaluated using a standardised tool. The 23 studies that met the inclusion criteria were categorised into subgroups depending on the type of comparison: hand hygiene product; skin condition; hand drying method. A 'best evidence synthesis' was utilised for classifying the evidence. RESULTS: Included studies provided evidence to support the use of alcohol-based hand rub as the preferred hand hygiene product. There was conflicting evidence for the use of medicated or plain soap, or any particular method to dry hands. CONCLUSIONS: Hand hygiene is a crucial component of risk management for both health care workers and their patients. It is important that hand hygiene practice is based on the best current evidence. As a result of a systematic review, evidence-based recommendations for hand hygiene for health care workers are proposed. PMID- 18425157 TI - A rare cause of early post-partum haematuria secondary to uterovesical fistula. PMID- 18425158 TI - Impacted radio-opaque glass in the oesophagus of a child. AB - Most ingested foreign bodies in the aero-digestive tracts in both children and adults are not radio-opaque, and as a result, a preoperative plain radiograph may not be helpful. However, incorporating radio-opaque markers into such potential foreign bodies like toys, beads, and dentures during manufacture (as illustrated in this case of a 6-year-old boy who ingested a piece of glass mirror which was easily seen on X-ray) would obviate this problem. While this may constitute some financial burden on manufacturers, the cost effectiveness on health management is non-negotiable. We suggest that appropriate authorities and regulatory bodies should enforce this through legislation. PMID- 18425159 TI - What is the state of health services research in New Zealand? AB - The article reflects on the current state of health services research (HSR) in New Zealand. A recent HSR conference held in Auckland highlighted a number of issues regarding the practice of HSR, suggesting that while there is some good research currently being conducted on issues pertaining to access, quality and costs of health services, more needs to be done. There is limited capacity in the public sector to conduct this work but barriers exist to commissioning academic and private researchers. Academic and private researchers have additional capacity, but often face constraints in producing policy-relevant research. We discuss ways that we might better coordinate and conduct HSR in the future. PMID- 18425160 TI - Medical image. Acute lower gastrointestinal bleeding: multidetector-row CT findings. PMID- 18425161 TI - Medical image. Portomesenteric venous gas and pneumatosis intestinalis. PMID- 18425162 TI - Alcohol and breast cancer. PMID- 18425163 TI - An update on tobacco smoking among New Zealand health care workers, the current picture, 2006. PMID- 18425164 TI - "We must discuss suicide", and response by Dr Beautrais. PMID- 18425165 TI - Barefoot running and walking: the pros and cons based on current evidence. PMID- 18425166 TI - Mid-infrared spectroscopic investigation of methylamines by a continuous-wave difference-frequency-generation-based system. AB - A laser spectroscopic system based on a cw difference-frequency generation source with a ratiometric multipass absorption detection scheme was employed for high resolution spectroscopic investigation of gas-phase monomethylamine (MMA), dimethylamine (DMA), and trimethylamine (TMA). Possible application of the system as a noninvasive human breath analyzer for renal and liver diseases is targeted. The system operates in the fundamental C-H stretch absorption region around 2740 2860 cm(-1). A detection sensitivity of 2 x 10(-6) cm(-1) Hz(-(1/2)) (for signal to-noise ratio SNR=1) is achieved, corresponding to detection limits of 900 ppb (parts in 10(9)) for MMA, 450 ppb for DMA, and 120 ppb for TMA in mixtures containing H2O and CO2 with concentrations of up to those present in human breath (2% and 5%, respectively). Future developments are discussed to further improve these detection limits that are currently still about 2 orders of magnitude higher than required for direct methylamine monitoring in human breath. PMID- 18425167 TI - Tight focusing of radially and azimuthally polarized vortex beams through a uniaxial birefringent crystal. AB - Under the approximation of small birefringence, the properties of radially and azimuthally polarized vortex beams tightly focused through a uniaxial birefringent crystal are investigated. The contour plots of intensity distribution near the focus and in the real focal plane are illustrated by performing numerical calculations. The dependence of the focal shift on numerical aperture and birefringence are analyzed. Moreover, the Strehl ratio in the real focal plane as a function of birefringence is also analyzed. It is revealed that the variation of birefringence has no influence on the focal shift and the Strehl ratio of azimuthally polarized vortex beams. PMID- 18425168 TI - Quasi-flat-top frequency-doubled Nd:glass laser for pumping of high-power Ti:sapphire amplifiers at a 0.1 Hz repetition rate. AB - A Nd:glass laser based on a novel design delivers up to 120 J energy pulses with a quasi-flat-top spatial profile at a 0.1 Hz repetition rate. The laser output is frequency-doubled with 50% efficiency and used to pump Ti:sapphire amplifiers. The developed design is perspective for use in the currently contemplated next step in ultra-high-intensity laser development. PMID- 18425169 TI - Formation of all fourteen Bravais lattices of three-dimensional photonic crystal structures by a dual beam multiple-exposure holographic technique. AB - We make use of a dual beam multiple-exposure (DBME) holographic technique for the formation of all 14 Bravais lattices of three-dimensional photonic crystal microstructures. For simplicity of experimental implementation, the DBME method has been modified such that, prior to each exposure, once the proper angle between the wave vectors of the interfering beams is chosen, a single axis rotation of the recording medium gives the desired results. The parameters required for the generation of the lattice structures have been derived by appropriate modification of interference of four noncoplanar beams (IFNB) analysis for corresponding implementation in the DBME technique, and the results have been verified by computer simulations. After giving a comparative study of the results with the IFNB method, recording geometries for the DBME approach are also proposed in order to realize all 14 Bravais lattices experimentally. PMID- 18425170 TI - Finite-difference-time-domain analysis of finite-number-of-periods holographic and surface-relief gratings. AB - The total-field-scattered-field formulation of the finite-difference time-domain method (FDTD) is used to analyze the diffraction of finite incident beams by finite-number-of-periods holographic and surface-relief gratings. Both second order and fourth-order FDTD formulations are used with various averaging schemes to treat permittivity discontinuities and a comparative study is made with alternative numerical methods. The diffraction efficiencies for gratings of several periods and various beam sizes, for both TE and TM polarization cases, are calculated and the FDTD results are compared with the finite-difference frequency-domain (FDFD) method results in the case of holographic gratings, and with the boundary element method results in the case of surface-relief gratings. Furthermore, the convergence of the FDTD results to the rigorous coupled-wave analysis results is investigated as the number of grating periods and the incident beam size increase. PMID- 18425171 TI - Dust absorption averaged over the Sahara inferred from moderate resolution imaging spectroradiometer. AB - We estimated the average dust single scattering albedo (omega(0)) over the Sahara using four years of moderate resolution imaging spectroradiometer data. The method employed is based on the theory that the critical surface reflectance (rho(c)) for which the reflectance at the top-of-the-atmosphere is not influenced by the variability of dust optical thickness depends on omega(0). The average dust absorption over the Sahara was estimated to be smaller than that previously reported in the literature, and this may be causing the cooling of the climate system. Our method enables one to estimate omega(0) from data with a variety of aerosol optical thickness values using samples in the vicinity of rho(c). Use of satellite data over large areas and for long periods enables quantification of the average dust absorption over the entire Sahara. PMID- 18425172 TI - 3x3 Mach-Zehnder interferometer with unbalanced differential detection for full range swept-source optical coherence tomography. AB - Quadrature interferometry based on 3x3 fiber couplers could be used to double the effective imaging depth in swept-source optical coherence tomography. This is due to its ability to suppress the complex conjugate artifact naturally. We present theoretical and experimental results for a 3x3 Mach-Zehnder interferometer using a new unbalanced differential optical detection method. The new interferometer provides simultaneous access to complementary phase components of the complex interferometric signal. No calculations by trigonometric relationships are needed. We demonstrate a complex conjugate artifact suppression of 27 dB obtained in swept-source optical coherence tomography using our unbalanced differential detection. We show that our unbalanced differential detection has increased the signal-to-noise ratio by at least 4 dB compared to the commonly used balanced detection technique. This is due to better utilization of optical power. PMID- 18425173 TI - Diffuse optical tomography guided quantitative fluorescence molecular tomography. AB - We describe a method that combines fluorescence molecular tomography (FMT) with diffuse optical tomography (DOT), which allows us to study the impact of heterogeneous optical property distribution on FMT, an issue that has not been systemically studied. Both numerical simulations and phantom experiments were performed based on our finite-element reconstruction algorithms. The experiments were conducted using a noncontact optical fiber free, multiangle transmission system. In both the simulations and experiments, a fluorescent target was embedded in an optically heterogeneous background medium. The simulation results clearly suggest the necessity of considering the absorption coefficient (mu(a)) and reduced scattering coefficient (mu'(s)) distributions for quantitatively accurate FMT, especially in terms of the accuracy of reconstructed fluorophore absorption coefficient (mu(a(x-->m))). Subsequent phantom experiments with an indocyanine green (ICG)-containing target confirm the simulation findings. In addition, we performed a series of phantom experiments with low ICG concentration (0.1, 0.2, 0.4, 0.6 and 1.0 microM) in the target to systematically evaluate the quantitative accuracy of our FMT approach. The results indicate that, with the knowledge of optical property distribution, the accuracy of the recovered fluorophore concentration is improved significantly over that without such a priori information. In particular absolute value of mu(a(x-->m) ) from our DOT guided FMT are quantitatively consistent with that obtained using spectroscopic methods. PMID- 18425174 TI - Study of process parameters on optical qualities for injection-molded plastic lenses. AB - Numerical simulations for mold-flow analysis and experimental measurements of injection-molded plastic lenses have been conducted for investigation of optical qualities, residual birefringence, and form accuracy resulting from various pertinent process conditions. First, residual birefringence distributions on the lens have been predicted and verified experimentally. Furthermore, full-scale factorial design of experiments was conducted to comprehend the influences of qualities, such as shear stresses, form accuracy, and volumetric deviation, on the measured primary or Seidel aberrations. In conclusion, residual birefringence induced by stresses represented by photoelasticity measurements agrees well with the numerical predictions and the experimental results indicate that the residual birefringence is mainly generated during the mold-filling stage. In addition, spherical aberration of the injection-molded plastic lenses is more sensitive to the pertinent qualities as compared to coma and astigmatism. PMID- 18425175 TI - Simultaneous detection of molecular oxygen and water vapor in the tissue optical window using tunable diode laser spectroscopy. AB - We report on a dual-diode laser spectroscopic system for simultaneous detection of two gases. The technique is demonstrated by performing gas measurements on absorbing samples such as an air distance, and on absorbing and scattering porous samples such as human tissue. In the latter it is possible to derive the concentration of one gas by normalizing to a second gas of known concentration. This is possible if the scattering and absorption of the bulk material is equal or similar for the two wavelengths used, resulting in a common effective pathlength. Two pigtailed diode lasers are operated in a wavelength modulation scheme to detect molecular oxygen ~760 nm and water vapor ~935 nm within the tissue optical window (600 nm to 1.3 mum). Different modulation frequencies are used to distinguish between the two wavelengths. No crosstalk can be observed between the gas contents measured in the two gas channels. The system is made compact by using a computer board and performing software-based lock-in detection. The noise floor obtained corresponds to an absorption fraction of approximately 6x10(-5) for both oxygen and water vapor, yielding a minimum detection limit of ~2 mm for both gases in ambient air. The power of the technique is illustrated by the preliminary results of a clinical trial, nonintrusively investigating gas in human sinuses. PMID- 18425176 TI - Sources and assumptions for the vicarious calibration of ocean color satellite observations. AB - Spaceborne ocean color sensors require vicarious calibration to sea-truth data to achieve accurate water-leaving radiance retrievals. The assumed requirements of an in situ data set necessary to achieve accurate vicarious calibration were set forth in a series of papers and reports developed nearly a decade ago, which were embodied in the development and site location of the Marine Optical BuoY (MOBY). Since that time, NASA has successfully used data collected by MOBY as the sole source of sea-truth data for vicarious calibration of the Sea-viewing Wide field of-view Sensor (SeaWiFS) and Moderate Resolution Imaging Spectroradiometer instruments. In this paper, we make use of the 10-year, global time series of SeaWiFS measurements to test the sensitivity of vicarious calibration to the assumptions inherent in the in situ requirements (e.g., very low chlorophyll waters, hyperspectral measurements). Our study utilized field measurements from a variety of sources with sufficient diversity in data collection methods and geophysical variability to challenge those in situ restrictions. We found that some requirements could be relaxed without compromising the ability to vicariously calibrate to the level required for accurate water-leaving radiance retrievals from satellite-based sensors. PMID- 18425177 TI - Single frame interferogram evaluation. AB - We present a simple and novel algorithm for the phase extraction from a single interferogram based on the spatial processing of interference patterns. This new evaluation procedure is suitable for application in environments where the presence of vibrations impedes the use of a classical phase-shifting interferometry scheme with multiple exposures. The algorithm does not require the introduction of a linear carrier as required in Fourier transform techniques. The addition of a carrier can be a significant drawback, e.g. in the case of wavefronts with strong aberrations where the minimum required linear carrier is not even resolved by the detector. The basic idea relies on the spatial application of a temporal phase-shifting algorithm and an iterative correction process to obtain an accurate reconstruction of the wavefront. The validity and performance of the proposed method is shown with numerical and experimental results. PMID- 18425178 TI - Multiple-wavelength digital holographic interferometry using tunable laser diodes. AB - Here we present multiple-wavelength digital holographic interferometry with a wide measurement range using laser diodes. Small wavelength differences can be easily realized by the wavelength tuning of laser diodes with injection current controls. A contour map of an object with a wide measurement range and a high sensitivity is demonstrated by combining a few contour maps with several measurement sensitivities. Synthetic wavelengths are calibrated using a known height difference. This alleviates the need to have high precise knowledge of the recording wavelengths. The synthetic wavelengths ranged from ~3 mm for high measurement sensitivity to ~4 cm for wide measurement range. An rms error of ~35 mum for a ~1 cm height measurement is shown. The measured profile of holographic interferometry agrees with a standard stylus instrument. PMID- 18425179 TI - Laser Doppler blood flow complementary metal oxide semiconductor imaging sensor with analog on-chip processing. AB - A 4 x 4 pixel array with analog on-chip processing has been fabricated within a 0.35 mum complementary metal oxide semiconductor process as a prototype sensor for laser Doppler blood flow imaging. At each pixel the bandpass and frequency weighted filters necessary for processing laser Doppler blood flow signals have been designed and fabricated. Because of the space constraints of implementing an accurate omega(0.5) filter at the pixel level, this has been approximated using the "roll off" of a high-pass filter with a cutoff frequency set at 10 kHz. The sensor has been characterized using a modulated laser source. Fixed pattern noise is present that is demonstrated to be repeatable across the array and can be calibrated. Preliminary blood flow results on a finger before and after occlusion demonstrate that the sensor array provides the potential for a system that can be scaled to a larger number of pixels for blood flow imaging. PMID- 18425180 TI - Stabilizing method for reflection interference contrast microscopy. AB - Reflection interference contrast microscopy (RICM) is a technique for measuring the shape and position of microscopic objects in solution; it has many biological and biophysical applications. Use of RICM for long-time acquisitions requires minimizing defocusing effects that are due to thermal and mechanical drift. We present a simple stabilizing method that accomplishes this using an image analysis-based linear focus function to establish feedback control of the focal position. While implementing this routine, we used RICM for independent measurement of the apparent fluctuation in the vertical position of an immobilized bead: the measured height had a standard deviation of 0.12 nm during a 45 min acquisition while under feedback control, demonstrating the high stability achievable with our approach. PMID- 18425181 TI - Spatial amplitude and phase modulation using commercial twisted nematic LCDs. AB - We present a method for full spatial phase and amplitude control of a laser beam using a twisted nematic LCD combined with a spatial filter. By spatial filtering we combine four neighboring pixels into one superpixel. At each superpixel we are able to independently modulate the phase and the amplitude of light. We experimentally demonstrate the independent phase and amplitude modulation using this novel technique. Our technique does not impose special requirements on the spatial light modulator and allows precise control of fields even with imperfect modulators. PMID- 18425182 TI - Optimal implementation of a microspectrometer based on a single flat diffraction grating. AB - An analytical model has been developed and applied to explore the limits in the design of a highly miniaturized planar optical microspectrometer based on an imaging diffraction grating. This design tool has been validated as providing the smallest possible dimensions while maintaining acceptable spectral resolution. The resulting planar spectrometer is composed of two parallel glass plates, which contain all components of the device, including a reflective slit and an imaging diffraction grating. Fabrication is based on microelectromechanical system technology and starts with a single glass wafer; IC-compatible deposition and lithography are applied to realize the parts in aluminum, which makes the microspectrometer highly tolerant for component mismatch. The fabricated spectrometer was mounted directly on top of an image sensor and takes up a volume of only 50 mm(3). The measured spectral resolution of 6 nm (FWHM) in the 100 nm operating wavelength range (600-700 nm) is in agreement with a model calculation. PMID- 18425183 TI - Shape reconstruction from gradient data. AB - We present a generalized method for reconstructing the shape of an object from measured gradient data. A certain class of optical sensors does not measure the shape of an object but rather its local slope. These sensors display several advantages, including high information efficiency, sensitivity, and robustness. For many applications, however, it is necessary to acquire the shape, which must be calculated from the slopes by numerical integration. Existing integration techniques show drawbacks that render them unusable in many cases. Our method is based on an approximation employing radial basis functions. It can be applied to irregularly sampled, noisy, and incomplete data, and it reconstructs surfaces both locally and globally with high accuracy. PMID- 18425184 TI - Low-loss waveguides fabricated in BK7 glass by high repetition rate femtosecond fiber laser. AB - For the first time femtosecond-laser writing has inscribed low-loss optical waveguides in Schott BK7 glass, a commercially important type of borosilicate widely used in optical applications. The use of a variable repetition rate laser enabled the identification of a narrow processing window at 1 MHz repetition rate with optimal waveguides exhibiting propagation losses of 0.3 dB/cm and efficient mode matching to standard optical fibers at a 1550 nm wavelength. The waveguides were characterized by complementary phase contrast and optical transmission microscopy, identifying a micrometer-sized guiding region within a larger complex structure of both positive and negative refractive index variations. PMID- 18425185 TI - Three-dimensional profiling using the Fourier transform method with a hexagonal grating projection. AB - We present three-dimensional profilometry based on triangulation in which a hexagonal pattern is projected on the object. To obtain an accurate result with a one-shot photographic image, the Fourier transform method and method of excess fraction are adopted. The three grating components of the hexagonal pattern are used. For compactness a new pattern projection scheme is introduced. The experimental results show that the constructed optical system works well for measuring the profile of a mannequin with a height resolution of ~ +/- 1 mm. PMID- 18425186 TI - Probing multilayer stack reflectors by low coherence interferometry in extreme ultraviolet. AB - We use low coherence interferometry to investigate the depth structure of a complex multilayer stack reflector. The probing instrument is an interferometer based on a Fresnel's bi-mirror illuminated by relatively wide-band synchrotron undulator light near 13.5 nm. Simulations clearly confirm that our test object generates two back propagated signals that behave as if reflected on two effective planes. First results in this spectral range may open the way to a new physical approach to extreme ultraviolet sample characterization in the form of line-scan optical coherence tomography. PMID- 18425187 TI - Wide-range sounding of free-tropospheric water vapor with a differential absorption lidar (DIAL) at a high-altitude station. AB - A differential absorption lidar (DIAL) system has been developed for the measurement of water vapor throughout the free troposphere [3 to 12 km above sea level (asl.)] with high vertical resolution varied from 50 m next to the ground to 300 m above an altitude of 10 km. The system was installed at the Schneefernerhaus high-altitude research station (2675 m asl., Zugspitze, Germany). The DIAL system is based on a tunable single-mode laser system with a high pulse energy of currently 250 mJ and a repetition rate of 20 s(-1). For lidar operation with energies typically between 100 mJ and 150 mJ and an integration time of 1000 s (10000 laser shots for both DIAL wavelengths) a vertical range of at least 10 km has been demonstrated even under dry conditions and during daytime, while daytime measurements up to 12 km have been possible under humid conditions. The system was intercompared with radiosondes, which suggests an agreement within 5% in a major part of the operating range. Further improvements are planned in the upper troposphere to approach the accuracy requirements needed in climate research. PMID- 18425188 TI - Three-flat test with plates in horizontal posture. AB - Measuring flats in the horizontal posture with interferometers is analyzed in detail, taking into account the sag produced by gravity. A mathematical expression of the bending is provided for a plate supported at three unevenly spaced locations along the edge. It is shown that the azimuthal terms of the deformation can be recovered from a three-flat measuring procedure, while the pure radial terms can only be estimated. The effectiveness of the iterative algorithm for data processing is also demonstrated. Experimental comparison on a set of three flats in horizontal and upright posture is provided. PMID- 18425189 TI - Multispectral camera and radiative transfer equation used to depict Leonardo's sfumato in Mona Lisa. AB - The technique used by Leonardo da Vinci to paint flesh tints--the sfumato--has never been scientifically depicted until now. From 100,000,000 reflectance spectra recorded on Mona Lisa, a virtual removal of the varnish is first obtained. A unique umber pigment is then identified in the upper layer and an exceptional maximum of the color saturation is underlined, both characteristics of a glaze technique. The modeling calling upon the radiative transfer equation confirms this maximum of saturation, the identification of an umber in the upper layer, and moreover underlines a mixture of 1% vermilion and 99% lead white in the base layer. Finally, the modeling, using the auxiliary function method, explains the spectacular maximum of saturation by the multiple scattering. PMID- 18425190 TI - Optimization of multilayer mirrors at 13.4 nm with more than two materials. AB - The design of multilayer mirrors with more than two materials is one of the key technologies for investigating lithography. We study a new procedure for optimizing multilayer mirrors of different combinations of materials at a wavelength of 13.4 nm. By adding Be and C layers in different orders to a Si/Mo stack, we have observed enhancement of the reflectivity and a reduction in the number of layers. The Luus-Jaakola optimization procedure has been implemented for the global optimization of the multilayer mirrors. With this algorithm it is not necessary to specify initially the number of layers present in a given design. PMID- 18425191 TI - Effects of sample polydispersity and beam profile on ellipsometric light scattering. AB - Ellipsometric light scattering (ELS) is shown to selectively extract the coherent scattering contribution representing the averaged properties of a particle ensemble. This property is essential for the previously reported [Erbe et al., Phys. Rev. E73, 031406 (2006)] high sensitivity of ELS to the refractive index profile at particle interfaces. Two mechanisms for coherence loss in ELS measurements are discussed: sample polydispersity and illumination by a Gaussian beam. Suitable experimental quantities for a distinction of coherent and incoherent scattering contributions are introduced. Furthermore, the application of the concepts to reflection ellipsometry at rough surfaces is discussed. PMID- 18425192 TI - Short coherence length produced by a spatial incoherent source applied for the Linnik-type interferometer. AB - An axial resolution created by a spatial incoherent source is investigated theoretically and experimentally for the Linnik-type interferometer. The axial resolution in interference microscopy depends on both the temporal coherence length of the source and the objective numerical aperture (NA). Here the problem is treated in a more general situation by considering the spatial and temporal coherence of the illumination source which may be important for deep coherence imaging application. The results show that the axial resolution is degraded at the depth much less by using the optimal spectral bandwidth of the incoherent source and high-NA objectives. PMID- 18425193 TI - Analysis of optically pumped equilateral triangular microlasers with three mode selective trenches. AB - Semiconductor laser devices based on triangular resonators can provide cheap, compact, and high performance optical sources for optical communications, computing, defense, and biological applications. I modify the original structure by introducing three trenches and analyze their effects on the electromagnetic modes propagating in the triangular cavity. I also analyze the coupling of light into single-mode waveguides. These analyses are conducted by using two dimensional finite difference time-domain methods. Results show that the introduction of such trenches can considerably reduce the quality factors of most of the modes, but one mode is not significantly degraded, providing nearly single mode operation. The effects of radiation losses are further investigated by introducing a photonic crystal shielding around the triangular structure. Finally I solve the rate equations to obtain the steady-state response for these structures. PMID- 18425211 TI - Synthesis, Solution and Solid State Structure of Titanium-Maltol Complex. AB - The reaction of Cp(2)TiCl(2) with two equivalents of maltol (3-hydroxy-2-methyl-4 pyrone) in water, at room temperature and pH of 5.4, leads to a complete replacement of Cp and chloride ligands affording, Ti(maltolato)(2)(OH)(2.) The complex has been characterized by IR, NMR and ESI-MS spectroscopic and cyclic voltammetry methods. In DMSO-d(6) solution, the complex shows two isomers in a ratio of 4:1, in which one OH signal can be identified per isomer. This suggests that in solution the complex is monomeric, most likely a chiral cis Ti(maltolato)(2)(OH)(2) and trans-Ti(maltolato)(2)(OH)(2). The monomeric nature of the complex (in water/methanol 1:1) was verified by ESI-MS spectroscopy, showing a parent peak at 329 m/z. Electrochemical behavior of Ti(maltolato)(2)(OH)(2) using cyclic voltammetry experiments showed the complex undergoes irreversible reduction in aprotic solvents. In D(2)O solution, at pH of 8.4, the (1)H NMR spectrum of the complex shows a mixture of monomer and tetramer Ti(IV)-maltol complexes in a ratio of 1:1. The crystallization of Ti(maltolato)(2)(OH)(2) at pH of 8.4 leads to the formation of [Ti(4)(maltolato)(8)(MU-O(4))]*18H(2)O. A single crystal of [Ti(4)(maltolato)(8)(MU-O(4))]*18H(2)O was analyzed by X-ray diffraction methods. The complex crystallizes in a monoclinic space group P2(1)/c with a = 12.617(4) A, b = 24.058(8) A, c = 22.686(7) A, beta= 97.678(4) degrees and V = 6824(4) A(3) for Z = 4. Solid state structure determination of the Ti-maltol complex showed to be tetrameric, containing two bridging oxides (in cis position) and two bidentate maltol ligands per titanium in a pseudo-octahedral coordination geometry. PMID- 18425212 TI - Design, Syntheses, and Characterization of a Sterically Encumbered Dioxo Molybdenum (VI) Core. AB - Dioxo-Mo(VI) complexes of general formula Tp*MoO(2)(p-SC(6)H(4)Dn) (6a-6c) (where Tp* = hydrotris(3,5-dimethyl-pyrazol-1-yl)borate and Dn= dendritic unit) have been synthesized and characterized by spectroscopy and mass spectrometry. (1)H NMR spectra of the metal complexes indicate that the C(s) local symmetry about the metal core does not change by the incorporation of dendritic functionality at the thiophenolato ring. Electrochemical data show ~20 mV change in the redox potential in the complexes with dendritic ligands suggesting a very small perturbation in the redox orbital, which is also supported by small changes in the electronic spectra. The peak-to peak separation (DeltaE(p)) increases from 125 mV in 6(a) to 240 mV in 6(c), suggesting sluggish electron transfer in molecules with larger dendritic ligands. PMID- 18425214 TI - Beyond Field Effect: Analysis of Shrunken Centroids in Normal Esophageal Epithelia Detects Concomitant Esophageal Adenocarcinoma. AB - BACKGROUND AND AIMS: Because of the extremely low neoplastic progression rate in Barrett's esophagus, it is difficult to diagnose patients with concomitant adenocarcinoma early in their disease course. If biomarkers existed in normal squamous esophageal epithelium to identify patients with concomitant esophageal adenocarcinoma, potential applications would be far-reaching. The aim of the current study was to identify global gene expression patterns in normal esophageal epithelium capable of revealing simultaneous esophageal adenocarcinoma, even located remotely in the esophagus. METHODS: Tissues comprised normal esophageal epithelia from 9 patients with esophageal adenocarcinoma, 8 patients lacking esophageal adenocarcinoma or Barrett's, and 6 patients with Barrett's esophagus alone. cDNA microarrays were performed, and pattern recognition in each of these subgroups was achieved using shrunken nearest centroid predictors. RESULTS: Our method accurately discriminated normal esophageal epithelia of 8/8 patients without esophageal adenocarcinoma or Barrett's esophagus and of 6/6 patients with Barrett's esophagus alone from normal esophageal epithelia of 9/9 patients with Barrett's esophagus and concomitant esophageal adenocarcinoma. Moreover, we identified genes differentially expressed between the above subgroups. Thus, based on their corresponding normal esophageal epithelia alone, our method accurately diagnosed patients who had concomitant esophageal adenocarcinoma. CONCLUSIONS: These global gene expression patterns, along with individual genes culled from them, represent potential biomarkers for the early diagnosis of esophageal adenocarcinoma from normal esophageal epithelia. Genes discovered in normal esophagus that are differentially expressed in patients with vs. without esophageal adenocarcinoma merit further pursuit in molecular genetic, functional, and therapeutic interventional studies. PMID- 18425215 TI - Periventricular leukomalacia in very low birth weight preterm neonates with high risk for neonatal sepsis. AB - OBJECTIVE: To investigate the association between periventricular leukomalacia (PVL) and neonatal sepsis in very low birth weight infants (VLBWI). METHODS: We studied VLBWI with a clinical suspicion of infection who had been born at our institution between the 1st of August, 2005 and the 31st of July, 2007. Children were excluded if they died before reaching 14 days, had malformations of the central nervous system or congenital infections. Ultrasound brain scans were carried out on the third day and weekly up until the sixth week of life or discharge. Periventricular leukomalacia was diagnosed by persistent diffuse periventricular hyperechogenecity for more than 7 days, or by periventricular cysts. The VLBWI were separated into two groups on the basis of the presence or absence of PVL. Sepsis was defined as clinical manifestation plus a positive culture. The Mann-Whitney, chi-square and t tests were applied followed by logistic regression. RESULTS: A total of 88 VLBWI were studied. Of these, 62 (70.5%) survived and 51 (57.8%) had PVL. Both groups were similar in terms of birth weight, gestational age, Apgar score, type of delivery, SNAPPE-II score, presence of necrotizing enterocolitis, persistent ductus arteriosus and deaths. Sepsis and mechanical ventilation were more common in the group with PVL (23.5 and 2.7%, p = 0.005; 86 and 59%, p = 0.004, respectively). Both of these were identified as, independent risk factors for PVL by logistic regression (p = 0.027 and 0.015, respectively). CONCLUSIONS: Chorioamnionitis has been defined as a risk factor for PVL. We have demonstrated that neonatal sepsis is also an important risk factor. We believe that the systemic inflammatory response is the principal factor involved in the etiopathogenesis of PVL among VLBWI. PMID- 18425213 TI - Parallels between pathogens and gluten peptides in celiac sprue. AB - Pathogens are exogenous agents capable of causing disease in susceptible organisms. In celiac sprue, a disease triggered by partially hydrolyzed gluten peptides in the small intestine, the offending immunotoxins cannot replicate, but otherwise have many hallmarks of classical pathogens. First, dietary gluten and its peptide metabolites are ubiquitous components of the modern diet, yet only a small, genetically susceptible fraction of the human population contracts celiac sprue. Second, immunotoxic gluten peptides have certain unusual structural features that allow them to survive the harsh proteolytic conditions of the gastrointestinal tract and thereby interact extensively with the mucosal lining of the small intestine. Third, they invade across epithelial barriers intact to access the underlying gut-associated lymphoid tissue. Fourth, they possess recognition sequences for selective modification by an endogenous enzyme, transglutaminase 2, allowing for in situ activation to a more immunotoxic form via host subversion. Fifth, they precipitate a T cell-mediated immune reaction comprising both innate and adaptive responses that causes chronic inflammation of the small intestine. Sixth, complete elimination of immunotoxic gluten peptides from the celiac diet results in remission, whereas reintroduction of gluten in the diet causes relapse. Therefore, in analogy with antibiotics, orally administered proteases that reduce the host's exposure to the immunotoxin by accelerating gluten peptide destruction have considerable therapeutic potential. Last but not least, notwithstanding the power of in vitro methods to reconstitute the essence of the immune response to gluten in a celiac patient, animal models for the disease, while elusive, are likely to yield fundamentally new systems level insights. PMID- 18425216 TI - Association of TGF-beta1, CD14, IL-4, IL-4R and ADAM33 gene polymorphisms with asthma severity in children and adolescents. AB - OBJECTIVE: To verify the association of transforming growth factor-beta1(TGF beta1) (C-509T and T869C), CD14 (C-159T), IL-4 (C-590T), IL-4R (ILe50Val) and ADAM33 (S_2) gene polymorphisms with asthma severity in a sample of patients with mild, moderate and severe persistent atopic asthma. METHODS: A clinical, laboratory, prospective study was performed in patients with persistent atopic asthma, compared to a control group at Hospital Universitario da Universidade Estadual de Campinas between 2006 and 2007. Analysis of the TGF-beta1 T869C gene polymorphism was performed using the technique polymerase chain reaction (PCR) + amplification refractory mutation system (ARMS). TGF-beta1 C-509T, CD14 C-159T, IL-4 C-590T, IL-4Ra ILe50Val, and ADAM33 S2 gene polymorphisms were detected by PCR and restriction enzyme. RESULTS: This study included 88 patients with persistent atopic asthma (27 mild, 23 moderate and 38 severe) and 202 healthy blood donors. As to T869C polymorphism (TGF-beta1), there was an association between the CC genotype and patients with severe asthma. There was no association in polymorphisms C-509T (TGF-beta1), C-590T (IL-4) and S_2 (ADAM33). When distribution of C-159T polymorphism genotype frequency (CD14) in severe asthma was compared with the control group, there was a significant result with the TT genotype. There was significant association of the Val/Val genotype (IL-4R) with mild asthma. CONCLUSION: Our results indicate that T869C (TGF-beta1), C-159T (CD14) and Val/Val (IL-4R) polymorphisms might be involved in modulation of asthma severity. PMID- 18425217 TI - [A historical view of rabies in Chile]. AB - In this review international and national historical features of rabies are presented remarking the start of preventive immunization in Chile, at year 1896, with the creation of the first service for rabies vaccination and la descentralizacion of preventive treatment from year 1929. Figures of human cases produced by this zoonosis between 1950 and 1986 are described, senalandose changes that occurred in local epidemiology of rabies since 1990, with an endemic pattern in bats and almost disappearance of canine rabies. PMID- 18425218 TI - [Rabies virus glycoprotein: structure, immunogenicity and pathogenic role]. AB - Rabies glycoprotein is the only exposed protein which is inserted in the viral lipidie envelope. This 65-67 kda protein is a N-glycosilated transmembrane protein forming trimers on the viral surface. It has been identified as the major pathogenicity determinant, playing a role in the budding, viral axonal transport during infection, apoptosis and immune evasion. It is also the major antigen responsible for the protective immune response and it is been used in commercial recombinant vaccines. Its structure, antigenicity and pathogenic role have been well studied, identifying main antigenic sites that have the responsibility for virulence, cellular receptors attachment and epitope acquisition. PMID- 18425219 TI - [Rabies in Chile: 1989-2005]. AB - A retrospective epidemiological study about epidemiology of rabies in Chile between years 1989 and 2005 was done. A data base of 39793 national registries of rabies samples was analyzed by means of statistical packages. Out of 39793 analyzed cases, 719 bats, 7 dogs, 7 cats, 1 bovine and 1 human were positive to rabies throughout the 17 years of this study. The statistical analysis established a significant increase in the proportions of positivity in bats, with predominance of variant 4 between the reservoirs. Given the complexity of the wild cycle of the rabies in Chile, it is necessary to maintain a program control of rabies, directed to educate people for a responsible possession of domestic animals, due to the risk of rabies transmission from bat to the susceptible species. PMID- 18425220 TI - [Morbid obesity: risks control]. PMID- 18425221 TI - Evaluation of risk factors in superobese patients submitted to conventional Fobi Capella surgery. AB - BACKGROUND: Obesity is one of the worlds greatest health problems. The Roux-en-Y gastric bypass is the gold standard treatment for severe obesity. Surgery in obese patients has an acceptable level of morbidity and mortality. The superobese patient, a subcategory of severe obese patients with a high surgical risk has not yet been analyzed as a group. METHODS: A retrospective and prospective cohort study was conducted enrolling 135 patients submitted to Roux-en-Y gastric bypass for treatment of severe obesity at the " Hospital das Clinicas" , Federal University of Pernambuco, Recife, PE, Brazil, between November 1997 and September 2003. The independent variables were possible risk factors of adverse outcomes: age, gender, weight, body mass index, diabetes, hypertension, hypercholesterolemia, sleep apnea, cardiopathy/coronariopathy, pneumopathy or any other co-morbidity. The dependent variables were major complications, minor complications and death. RESULTS: Diabetes (RR = 1.6 and CI = 1.02-2.40) and sleep apnea (RR = 1.8 and CI = 1.18-2.64) were associated to minor complications. Cardiopathy/coronariopathy were associated with major complications (RR = 5.42 and CI = 1.22-2.40) and death (RR = 16.25 and CI = 3.00-87.95). BMI >or=55 kg/m2 was associated with minor complications (RR = 1.58 and CI = 1.04-2.40), major complications (RR = 3.17 and CI = 1.03-9.80) and death (P = 0.007). After logistic regression, the body mass index >55 kg/m2 remained as a strong risk factor of death (OR = 3.6 and CI = 1.05-12.32). CONCLUSIONS: The body mass index >or=55 kg/m2 was the main risk factor for severe complications and death. Other risk factors affecting the outcome were diabetes, obstructive sleep apnea and cardiopathy/coronariopathy. PMID- 18425222 TI - Duplex sonography study in schistosomiasis portal hypertension: characterization of patients with and without a history of variceal bleeding. AB - BACKGROUND: Presinusoidal portal hypertension with frequent episodes of upper gastrointestinal variceal bleeding are hallmarks of hepatosplenic Mansons schistosomiasis; a clinical form that affects about 5% of Brazilians who are infected by Schistosoma mansoni. AIMS: To evaluate duplex sonography findings in patients with hepatosplenic Mansons schistosomiasis with and without upper gastrointestinal variceal hemorrhage. METHODS: A cross-sectional study was performed whereby 27 consecutive patients with hepatosplenic Mansons schistosomiasis were divided into two groups: group I (six men and six women; mean age 48.7 years) with a past history of bleeding and group II (four men and eight women; mean age 44.7 years) without a past history of upper gastrointestinal bleeding, underwent duplex sonography examination. All patients underwent the same upper gastrointestinal endoscopy and laboratory examinations. Those with signs of mixed chronic liver disease or portal vein thrombosis (three cases) were excluded. RESULTS: Group I showed significantly higher mean portal vein flow velocity than group II (26.36 cm/s vs 17.15 cm/sec). Although, as a whole it was not significant in all forms of collateral vessels (83% vs 100%), there was a significantly higher frequency of splenorenal collateral circulation type in group II compared with group I (17% vs 67%). The congestion index of the portal vein was significantly lower in group I than in group II (0.057 cm vs 0.073 cm/sec). CONCLUSION: Our duplex sonography findings in hepatosplenic Mansons schistosomiasis support the idea that schistosomotic portal hypertension is strongly influenced by overflow status, and that collateral circulation seems to play an important role in hemodynamic behavior. PMID- 18425223 TI - Endoscopic ultrasound-guided endoscopic transmural drainage of pancreatic pseudocysts. AB - BACKGROUND: Surgery is the traditional treatment for symptomatic pancreatic pseudocysts, but the morbidity is still too high. Minimally invasive endoscopic approaches have been encouraged. AIMS: To evaluate the efficacy of endoscopic ultrasound-guided endoscopic transmural drainage of pancreatic pseudocysts. METHODS: From January, 2003 to August, 2006, 31 consecutive symptomatic patients submitted to 37 procedures at the same endoscopic unit were retrospectively analysed. Chronic and acute pancreatitis were found in, respectively, 17 (54.8%) and 10 (32.3%) cases. Bulging was present in 14 (37.8%) cases. Cystogastrostomy or cystoduodenostomy were created with an interventional linear echoendoscope under endosonographic and fluoroscopic control. By protocol, only a single plastic stent, without nasocystic drain, was used. Straight or double pigtail stents were used in, respectively, 22 (59.5%) and 15 (40.5%) procedures. RESULTS: Endoscopic ultrasound-guided transmural drainage was successful in 29 (93.5%) patients. Two cases needed surgery, both due to procedure-related complications. There was no mortality related to the procedure. Twenty-four patients were followed-up longer than 4 weeks. During a mean follow-up of 12.6 months, there were six (25%) symptomatic recurrences due to stent clogging or migration, with two secondary infections. Median time for developing complications and recurrence of the collections was 3 weeks. These cases were successfully managed with new stents. Complications were more frequent in patients treated with straight stents and in those with a recent episode of acute pancreatitis. CONCLUSIONS: Endoscopic transmural drainage provides an effective approach to the management of pancreatic pseudocysts. PMID- 18425224 TI - [Endoscopic ultrasound in the preoperative staging of gastric cancer: correlation with the surgical and/or pathological findings]. AB - BACKGROUND: Gastric adenocarcinoma is the major cause of death by cancer in Brazil. For the planning of the treatment and evaluation of the prognosis, the preoperative staging according to the TNM classification is very important. AIM: To evaluate the results of endoscopic ultrasound for the T and N categories. METHODS: We examined 30 patients with gastric adenocarcinoma in the period of 1 year. We used a Olympus GIF UM-20. After positioning in the third portion of the duodenum, we started to evaluate the lymph node stations 16, 13, 12, 6, 5, 4, 3, 8, 7, 9, 10, 11, 1 and 2, using the frequency of 7.5 MHz. The depth of the tumor in the gastric wall was evaluated with the frequencies of 7.5 and 12 MHz. The results T and N were compared with the conclusive findings. In the unresectable cases, the correlation was made by the macroscopic findings. RESULTS: We performed 16 total gastrectomies, 7 subtotal gastrectomies, 5 exploratory laparotomies and 2 laparoscopies. For T category, endoscopic ultrasound results were correct in 25/30 cases (83.3%). All the other 16.7% were overestimated. The accuracy varied from 90% to 96.7% according to the subcategories T1 to T4. For the N category, endoscopic ultrasound results were correct in 23/30 cases (76.7%). Sixteen point six percent were underestimated and 6.7% overestimated. The accuracy varied from 76.7% to 90%, in agreement with the subcategories N1 to N3. CONCLUSION: There was a clear correlation between endoscopic ultrasound and the surgical and pathological findings in the evaluation of T and N categories in patients with gastric adenocarcinoma. PMID- 18425225 TI - Gastric cancer patients treated by a general or gastric cancer surgical team: a comparative study. AB - BACKGROUND: Although gastric cancer has been decreasing in incidence in many countries, it is still the second most common cause of cancer deaths worldwide. Its prognosis is poor and depends, among other factors, on early diagnosis as well as on surgeon expertise. AIM: To compare the outcomes of gastric cancer patients treated at a university hospital by a general surgical team and later on by a gastric cancer surgical team. METHODS: Gastric cancer patients were separated into two groups according to whether they were treated by a general surgical team (group 1, n = 136; 1984 to 1993) or by gastric cancer team (group 2, n = 149; 1994 to 2003). Clinical and pathologic features and survival rates were assessed. RESULTS: During a 20-year period, a decreased number of patients underwent surgical resection in the second period (94% vs 86%), a greater number of upper gastrointestinal endoscopies were performed resulting in an increased number of tumors diagnosed as stage I (5% vs 22%). Also, D2 gastrectomies were more frequently performed instead of D0 gastrectomies and negative surgical margins were adequate. Mortality decreased from 9% to 6% in group 1 and 2, respectively and adjuvant therapy has been considered. CONCLUSION: Surgical specialized units for gastric cancer are necessary if better results are to be expected since this approach definitely provides better patient care. PMID- 18425226 TI - Prevalence of hepatopulmonary syndrome in patients with decompensated chronic liver disease and its impact on short-term survival. AB - BACKGROUND: Hepatopulmonary syndrome is reported to occur in 4% to 32% of the patients with chronic liver disease and is associated with poor liver function and shortened patient survival before and after liver transplantation. AIMS: To assess the frequency of hepatopulmonary syndrome in Brazilian patients with decompensated chronic liver disease and to investigate its impact on patient survival. METHODS: One hundred and thirty patients (101 males, mean age 61 +/- 12 years) with decompensated chronic liver disease were evaluated for the presence of hepatopulmonary syndrome. The diagnosis of hepatopulmonary syndrome was considered in the presence of alveolar arterial oxygen gradient of more than 15 mm Hg and of pulmonary vascular dilatation assessed by contrast enhanced echocardiography. RESULTS: Hepatopulmonary syndrome was observed in 21 (16%) patients. The presence of hepatopulmonary syndrome was significantly associated with severity of liver disease assessed by the MELD (Model for End-Stage Liver Disease) score, but not with in hospital mortality after admission due to decompensated chronic liver disease. CONCLUSIONS: Hepatopulmonary syndrome occurs in 16% of patients with chronic liver disease and is associated with disease severity according to the MELD score. Short term mortality following decompensation of chronic liver disease was not associated with hepatopulmonary syndrome. PMID- 18425227 TI - [Vascularization in hepatic cirrhosis: an immunohistochemical study on necropsies]. AB - BACKGROUND: Fibrosis has been the most cited variable in cirrhosis, but major alterations in hepatic vascularization have been pointed as basic elements in the physiopathology of the illness and its complications as portal hypertension, hepatic failure and hepatocellular carcinoma. METHODS: The present study aims at assessing microvascular density in 35 cases of necropsies of cirrhotic patients by immunohistochemical detection of endothelial marker CD34, comparing semi quantification with morphometric quantitative method, also searching for a possible relation of vascular alterations with the main causal agents, injury patterns and major clinical complications. RESULTS: A significant association was detected between semi-quantitative and quantitative approach of microvessel density in parenchyma, but not in septa. No significant association was detected between neovascularization and any specific clinical complication of cirrhosis. Under our standpoint, the main achievement of the present study was the demonstration that the vascular neoformation in hepatic parenchyma is significantly higher in cirrhosis associated with chronic hepatitis than in cirrhosis resulting from steatohepatitis. CONCLUSION: These findings require further clinical studies to assess the hypothesis that the rearrangement of liver microcirculation through the detection of CD34 might be relevant in prognostic assessment of cirrhotic patients. PMID- 18425228 TI - Prevalence of Barrett's esophagus in individuals without typical symptoms of gastroesophageal reflux disease. AB - BACKGROUND: Barretts esophagus, the major risk factor for esophageal adenocarcinoma, is detected in approximately 10%-14% of individuals submitted to upper endoscopy for the assessment of gastroesophageal reflux disease related symptoms. Prevalence studies of Barretts esophagus in individuals without typical symptoms of gastroesophageal reflux disease have reported rates ranging from 0.6% to 25%. AIM: To determine the prevalence of Barretts in a Brazilian population older than 50 years without typical symptoms of gastroesophageal reflux disease. METHODS: A total of 104 patients (51 men), mean age of 65 years, with an indication for upper endoscopy but without symptoms of heartburn and/or acid regurgitation (determined with a validated questionnaire) were recruited. Subjects submitted to upper endoscopic examination in the last 10 years or using antisecretory medication (proton pump inhibitors) during the last 6 months were not included. Methylene blue chromoscopy was performed during the endoscopic exam to facilitate identification of the metaplastic epithelium. RESULTS: Barretts esophagus was diagnosed endoscopically and confirmed by histology in four patients, all of them males. The metaplastic segment was short (less than 3 cm) and free of dysplasia in all patients. The prevalence of Barretts esophagus was 7.75% in the male population and 3.8% in the general population studied. CONCLUSION: Due to the low prevalence of Barretts esophagus found in the present study, associated with the finding of short-segment Barretts esophagus in all cases diagnosed and the absence of dysplasia in the material analyzed, endoscopic screening for Barretts esophagus in patients above the age of 50 without the classical symptoms of gastroesophageal reflux disease is not indicated for the Brazilian population. PMID- 18425229 TI - Chronic constipation - the role of clinical assessment and colorectal physiologic tests to obtain an etiologic diagnosis. AB - BACKGROUND: Diagnosis of subtypes of chronic constipation has been considered difficult to achieve even in specialized centers. Although colorectal physiologic tests have brought an important contribution, it remains unclear in which patients these tests should be indicated for. AIMS: This study aims to establish a differential diagnosis for chronic constipation cases using clinical assessment and physiologic tests and to identify clinical parameters that could predict which patients need physiologic tests. METHODS: One hundred and seventy nine patients (83% females; mean age, 45) with chronic constipation according to Rome II criteria were initially treated by dietary advice and functional reeducation and those unresponsive (110 or 61.5%) were submitted to colonic transit time, defecography, anorectal manometry and electromyography, as needed. RESULTS: A differential diagnosis was achieved in 63.6% of patients tested. However, 61.5% of 179 patients with chronic constipation (69 with no need to tests and 40 with normal tests) have etiologic diagnosis established only on clinical basis. Irritable bowel syndrome (32%), pelvic floor dysfunction (29%) and functional constipation due to faulty diet and life style habits (22%) were the main causes of chronic constipation. Alternating constipation and nausea/vomiting were symptoms significantly related to the diagnosis of irritable bowel syndrome; younger age, larger intervals between bowel movements, occurrence of fecal impaction and necessity of enema were related to the diagnosis of non-chagasic megacolon and digital assistance to evacuate and large rectocele or spastic pelvic floor on rectal exam were associated to pelvic floor dysfunction. Patients with long-standing constipation, fecal impaction, abdominal pain not eased after defecation, necessity for enemas, digital assistance and evidence of rectocele tended to be in need for physiologic tests to define the cause of chronic constipation. CONCLUSIONS: The etiologic diagnosis of chronic constipation can be achieved in most of patients on a clinical basis and some symptoms may be significantly related to specific diagnoses. Indications for physiologic tests should be based on specific clinical parameters. PMID- 18425230 TI - [Fiber intake and colonic transit time in functional constipated patients]. AB - BACKGROUND: Patients with functional constipation presenting no response to treatment using fibers supplement represents important clinical issue. AIMS: To evaluate the relations among the amount of ingested fiber, the constipation intensity and the colonic transit time in patients with functional constipation. METHODS: We evaluated 30 patients, presenting no response to treatment using fibers supplement, and 18 healthy volunteers conducting individual inquiry into fibers intake, constipation intensity and the total and segmental colonic transit evaluation using radiopaque markers. RESULTS: In the constipated, despite the good level of fiber intake (26.3 +/- 12.9 g, constipated x 9.3 +/- 5,2 g, control), the symptoms of constipation was serious (score = 21.3 +/- 4.07). Mean total colonic transit was 58.8h. The colonic transit was slower in the constipated group (41.0 +/- 22.8 hours, constipated x 21.8 +/- 18.5h, control). In constipated patients with slow colonic transit (>58.8h) there were colonic inertia (eight), outlet constipation (one) and slow transit in left colon (one), and among constipated patients with normal colonic transit (<58.8h), there were isolated slow transit, in the right colon (nine), left colon (three) and in the rectosigmoid segment (eight). There were no relation among the amount of ingested fiber, constipation intensity and the colon transit. CONCLUSIONS: In the functional constipation the gravity of symptoms does not depend only on the dietary fibers intake, which is not the only responsible for the differences in the colonic transit. The colonic transit can differentiate normal from constipated patients and, among them, those with altered transit that demand approaches distinct of fiber supplementation. PMID- 18425231 TI - Application of the Vienna classification for Crohn's disease to a single center from Brazil. AB - BACKGROUND: Crohn's disease is a chronic inflammatory disorder with diversity on its clinical presentation that may be observed from the varying age of onset of symptoms to the site of occurrence of the illness. There is a need for a replicable and uniform description of the disease allowing a comparison between distinct study populations. The 1998 Vienna classification characterizes patients according to three clinical aspects: age at diagnosis, location and disease behavior. AIM: To describe Crohn's disease in patients from a reference center of Salvador, BA, Brazil according to the Vienna classification. METHODS: Between January and October of 2005, patients (n = 47) having at least one endoscopic and radiological examination of the intestine participated in this study. RESULTS: Most of the participants had the diagnosis of the disease when they were younger than 40 years old (70.2%) while an ileocolic location (38.3%) and the penetrating form (46.8%) were the most prevalent clinical presentation. The restricted location of the ileum (L1) was more frequent in nonstricturing, nonpenetrating disease (B1) while the ileocolic disease (L3) was more associated with the penetrating behavior (B3). CONCLUSION: In this study, differently from the first description of the Vienna classification, the large number of patients presenting a complicated stage of the disease can be attributed to the fact that it was carried out at a reference center, where many patients present with the disease at an advanced stage. PMID- 18425232 TI - Prevalence of celiac disease among first degree relatives of Brazilian celiac patients. AB - BACKGROUND: Several studies have shown that celiac disease, an autoimmune disorder that occurs in genetically susceptible individuals, is highly prevalent among relatives of celiac patients. AIM: To determine the prevalence of celiac disease in a group of first degree relatives of Brazilian celiac patients. METHODS: First degree relatives of celiac patients attending the Brasilia University Hospital Pediatric Gastroenterology Outpatient Clinic or the Celiac Disease Investigation Center, Brasilia, DF, Brazil, between March 2001 and November 2004 were invited to undergo serological screening for celiac disease applying the IgA anti-endomysium antibody test (IgA-EMA). All positive IgA-EMA sera underwent a second screening using the IgA anti-tissue transglutaminase antibodies test. Duodenal or small intestinal biopsies were performed in all subjects positive to serological testing. Biopsy samples were classified as type (O) normal, (I) infiltrative, (II) infiltrative hyperplastic, (III) flat destructive, and (IV) atrophic hypoplastic. The final diagnosis was ascertained in subjects showing positive serological tests and a grade I to III small intestinal lesion. RESULTS: Nine new cases of celiac disease were found among the 188 first degree relatives tested (4.8%). CONCLUSION: The present study confirms the high prevalence of celiac disease among first degree celiac patients relatives and reinforces the need of extensive diagnostic screening in this specific group. PMID- 18425233 TI - [Central inferior bisegmentectomy (S4B+S5) for gallbladder carcinoma treatment: a series of seven resectable cases]. AB - BACKGROUND: Despite its rarity, gallbladder cancer is an aggressive type of neoplasia with a very poor prognosis. The best resection for oncological purposes continues to be right hepatectomy extended to segment IV. However, bisegmentectomy IV-V is becoming an interesting alternative because of greater preservation of the parenchyma. AIM: To report the early and late results from bisegmentectomy IV-V in cases of carcinoma of the gallbladder. METHODS: A series of seven cases of invasive carcinoma is presented (six women and one man). These patients underwent bisegmentectomy IV-V at the General Surgery Service of the Teaching Hospital of the ABC Medical School, Santo Andre, SP, Brazil. The study was conducted between 2002 and 2006. The patients ages ranged from 52 to 72 years. The diagnosis was preoperative (radiological) in five cases, which were all confirmed by intraoperative frozen-tissue examination, while in two cases the diagnosis was postoperative, following open cholecystectomy. RESULTS: The duration of the operation ranged from 180 to 340 minutes. The quantity of intraoperative bleeding ranged from 200 to 1500 mL. There were two major complications but no mortality. Six patients did not present any recurrence over the course of 3 to 30 months of follow-up. CONCLUSION: Bisegmentectomy IV-V may constitute a curative surgical alternative for treating gallbladder cancer. This procedure presents acceptable morbidity and mortality. PMID- 18425234 TI - [Evaluation of the event-related potential (ERP-P300) in patients with hepatic cirrhosis without encephalopathy]. AB - BACKGROUND: In hepatic cirrhosis structural liver alterations occur leading to the loss of the organ functions with neuro-psychiatric consequence, as cognitive dysfunctions. One of the most effective ways of objectively evaluating cognition is to measure electrophysiological activity in the central nervous system trough event-related potentials (ERP-P300). AIM: To assess the value of the event related potential (ERP) in order to determine cognitive disturbances in patients with liver cirrhosis and to assist in the diagnosis of minimal hepatic encephalopathy. METHODS: Fifty patients with liver cirrhosis were selected, without clinical symptoms of hepatic encephalopathy and 35 healthy volunteers, matched by sex and age. The patients were submitted to clinical-neurological and laboratorial examination. The ERP-P300 was performed by the two groups to determine cognitive disturbances. RESULTS: The study showed significant differences between the ERP-P300 latency averages of the two groups. CONCLUSION: The ERP-P300 is simple to use and depends on controllable variables. It is also easy to reproduce and, when properly used, can be useful both to determine cognitive disturbances in patients with hepatic cirrhosis and to assist in minimal hepatic encephalopathy diagnosis. PMID- 18425235 TI - Morphine does no promote esophageal carcinogenesis in rats exposed to diethylnitrosamine. AB - BACKGROUND: The high incidence of esophageal cancer in the north of Iran has been associated to the consumption of opium and exposure to nitrosamines. Diethylnitrosamine has an established potential of producing experimental cancer in the esophagus and liver. AIM: To evaluate by histopathology the effect of oral administration of morphine and diethylnitrosamine during 23 weeks on the hepatic and esophageal carcinogenesis on 176 rats. METHODS: We divided the rats into the following groups: Morph: morphine; Den: diethylnitrosamine; Den+morph: Den and morphine in the same solution; Den/morph: Den and morphine in different solutions and days. RESULTS: Morphine did not promote neoplasias. The highest neoplastic incidents were found: a) in the esophagus, Den in relation to Den/morph and Den+morph (71.1%, 55.8%, and 50.0%); b) in the liver, Den and Den/morph in relation to Den+morph (73.8%, 81.4%, and 40.9%); c) higher incident of hepatic neoplasia than esophageal in Den/morph (81.4% and 55.8%). Different doses of diethylnitrosamine were ingested among the groups Den, Den/morph, and Den+morph, respectively 2.9, 2.8, and 2.3 mg/kg/day. CONCLUSIONS: These results show that the morphine did not promote esophageal carcinogenesis and may have stimulated the hepatic metabolism of the first pass of the carcinogen. PMID- 18425236 TI - [Effect of partially hidrolized guar-gum in the treatment of functional constipation among hospitalized patients]. AB - The effect of hydrolyzed partially guar-gum was evaluated in the treatment of functional constipation among hospitalized patients. Following a randomized blind controlled-trial 64 adults were allocated to two groups: one received daily high fiber diet (approximately 30 g) and the other similar diet plus 10 g of hydrolyzed partially guar-gum, during 15 days. Dietary fiber reduced functional constipation by 78.0%. Hydrolyzed partially guar-gum did not show any additional effect in defecation frequency, fecal consistence, need of laxative drug use, although a reduction in bowel complaints. Dietary fiber may be used in the treatment of functional constipation. However the therapeutic role of hydrolyzed partially guar-gum should be further investigated. PMID- 18425237 TI - The BOR (Brazilian Oral Research) is changing its submission instruction. PMID- 18425238 TI - Physicochemical characterization of two deproteinized bovine xenografts. AB - Calcium phosphate salts, or more specifically hydroxyapatite, are products of great interest in the fields of medical and dental science due to their biocompatibility and osteoconduction property. Deproteinized xenografts are primarily constituted of natural apatites, sintered or not. Variations in the industrial process may affect physicochemical properties and, therefore, the biological outcome. The purpose of this work was to characterize the physical and chemical properties of deproteinized xenogenic biomaterials, Bio-Oss (Geistlich Biomaterials, Wolhuser, Switzerland) and Gen-Ox (Baumer S.A., Brazil), widely used as bone grafts. Scanning electron microscopy, infrared region spectroscopy, X-ray diffraction, thermogravimetry and degradation analysis were conducted. The results show that both materials presented porous granules, composed of crystalline hydroxyapatite without apparent presence of other phases. Bio-Oss presented greater dissolution in Tris-HCl than Gen-Ox in the degradation test, possibly due to the low crystallinity and the presence of organic residues. In conclusion, both commercial materials are hydroxyapatite compounds, Bio-Oss being less crystalline than Gen-Ox and, therefore, more prone to degradation. PMID- 18425239 TI - Assessment in vitro of brushing on dental surface roughness alteration by laser interferometry. AB - Noncarious cervical lesions (NCCLs) are considered to be of multifactorial origin, normally associated with inadequate brushing. This study assessed the influence in vitro of simulated brushing on NCCL formation. Fifteen human premolars were submitted to brushing in the cementoenamel junction region, using hard-, medium- and soft-bristled brushes associated with a toothpaste of medium abrasiveness under a 200 g load, at a speed of 356 rpm for 100 minutes. The surface topography of the region was analyzed before and after brushing, by means of a laser interferometer, using "cut-off" values of 0.25 and considering roughness values in mm. The initial roughness (mm) results for dentin (D / bristle consistency: 1--soft, 2--medium and 3--hard) were as follows: (D1) 1.25 +/- 0.45; (D2) 1.12 +/- 0.44; (D3) 1.05 +/- 0.41. For enamel (E / bristle consistency: 1--soft, 2--medium and 3--hard), the initial results were: (E1) 1.18 +/- 0.35; (E2) 1.32 +/- 0.25; (E3) 1.50 +/- 0.38. After brushing, the following were the values for dentin: (D1) 2.32 +/- 1.99; (D2) 3.30 +/- 0.96; (D3) Over 500. For enamel, the values after brushing were: (E1) 1.37 +/- 0.31; (E2) 2.15 +/ 0.90; (E3) 1.22 +/- 0.47. Based on the results of the ANOVA and Tukey statistical analyses (a = .05) it was concluded that soft, medium and hard brushes are not capable of abrading enamel, whereas dentin showed changes in surface roughness by the action of medium- and hard-bristled brushes. PMID- 18425240 TI - Evaluation of the risk of a stripping perforation with Gates-Glidden drills: serial versus crown-down sequences. AB - The aim of this study was to evaluate the remaining dentine/cementum thickness using Gates-Glidden burs in serial and crown-down sequences and to observe which of the two sequences is the safest for preparing mesial roots of molars. Thirty six left and right human mandibular first molars were selected. Standard access cavities were made and initially explored with Flexofiles sizes 10 and 15 until the tip was visible at the apex. The teeth were embedded in a muffle specially developed for this study using a PVC tube with two parallel metal rods in its lid. Each tooth-block was sectioned 3 mm apically to the furcation using a low speed saw with a diamond disc. The tooth-block was examined under a microscope and an initial image was captured by a digital video system with 8 X and 12 X magnifications. Finally, the tooth-blocks were reassembled in the muffle so that the canals could be instrumented. After instrumentation the area of each mesial canal as well as the smallest distance to the root furcation were measured again. The mesio-buccal canals (crown-down order) and the mesio-lingual canals (serial sequence) presented an average area of 0.46 +/- 0.16 mm(2) and 0.88 +/- 0.27 mm(2) (P < 0.01), respectively. The mean values of the smallest distance to the furcation for the mesio-buccal and mesio-lingual canals were 0.66 +/- 0.19 mm and 0.39 +/- 0.13 mm (P < 0.01), respectively. The remaining dentine/cementum thickness using Gates-Glidden burs was greater in the crown-down sequence than in the serial sequence. PMID- 18425241 TI - The effect of endodontic materials on the optical density of dyes used in marginal leakage studies. AB - The aim of this study was to determine the effect of the exposure of different endodontic materials to different dye solutions by evaluating the optical density of the dye solutions. Seventy-five plastic tubes were filled with one of the following materials: AH Plus, Sealapex, Portland cement, MTA (Angelus and Pro Root) and fifteen control plastic tubes were not. Each specimen of material and control was immersed in a container with 1 ml of each dye solution. A 0.1 ml-dye solution aliquote was removed before immersion and after 12, 24, 48 and 72 hours of each specimen immersion to record its optical density (OD) in a spectrophotometer. Statistical analysis was performed with ANOVA and Tukey tests (5%). No significant difference was found among any of the solution OD values for AH Plus cement. Portland cement promoted different OD values after 12 hours of immersion. MTA-Angelus cement presented different OD values only for 2% rhodamine B and the MTA-Pro Root cement presented different OD values in all 2% rhodamine B samples. Sealapex cement promoted a reduction in the India Ink OD values. Dye evaluation through OD seems to be an interesting method to select the best dye solution to use in a given marginal leakage study. PMID- 18425242 TI - Prevalence of bruxism and emotional stress and the association between them in Brazilian police officers. AB - This study aimed to assess the prevalence of bruxism and emotional stress in Brazilian police officers, due to exposure to stressful situations, and to assess the relationship between the type of work done by a police officer and the presence of bruxism and emotional stress. A cross-sectional study was conducted at the Military Police of the State of Sao Paulo, Campinas, SP, Brazil. The final sample included 394 male police officers (mean age = 35.5 years). Bruxism was diagnosed by the presence of aligned dental wear facets associated with the presence of one of the following signs or symptoms: self-report of tooth grinding, painful sensitivity of the masseter and temporal muscles, discomfort in the jaw musculature upon waking. The Stress Symptoms Inventory (SSI) was applied to evaluate emotional stress. The type of work done by the police was classified as organizational or operational, the latter being assumed as the more stressful since it exposes the police officer to life risk. The results showed a prevalence of bruxism of 50.2% and a prevalence of emotional stress of 45.7%. The Chi-square test indicated an association between stress and bruxism (P < .05). No significant association was found between emotional stress and type of work (P = .382) or between bruxism and work activity (P = .611). It could be concluded that emotional stress was associated with bruxism, independently of the type of work done by police officers. PMID- 18425243 TI - Relationship between oral health and its impact on quality of life among adolescents. AB - The aim of this study was to assess oral health status and its relationship with quality of life. A household population, cross-sectional study was carried out; participants were between 15 and 17 years of age (n = 247) and were examined by two calibrated dentists. Socio-economic status was classified according to ANEP ABIPEME criteria. Clinical examinations to observe DMFT, CPI and Dean indices were performed as per WHO criteria. The Significant Caries Index (SiC) was used to evaluate polarization of the occurrence of caries among participants of the tercile with higher DMF-T. The OHIP instrument was used to measure quality of life. The Spearman and Mann-Whitney tests were used for assessing correlations (5% significance level). Examinations were carried out in 117 (47.37%) females and in 130 (52.63%) males. Of the examined participants, 45.75% were classified as belonging to socio-economic class C. Caries occurrence was observed in 218 subjects (88.26%); the mean DMFT was 5.40. The SiC index was 9.97. Almost half (47.77%) of the participants examined did not present sextants affected by periodontal disease. Of the participants examined, 80.16% presented absence of fluorosis. The mean OHIP was 3.95. The following correlations were observed: a positive and statistically significant correlation between the highest score in the OHIP and decayed teeth; a positive correlation with threshold significance between OHIP and DMFT; an inverse correlation between intact teeth and OHIP; and a positive and non statistically significant correlation between SiC and OHIP (correlation coefficient = 0.13, p = 0.245). Association between the mean OHIP and the terciles was not significant (p = 0.146); there were also no associations between periodontal condition and OHIP nor were there associations between the presence of fluorosis and mean OHIP. PMID- 18425244 TI - In vivo qualitative analysis of the biocompatibility of different cyanoacrylate based adhesives. AB - Cyanocrylates have been widely used in the medical and dental fields for several years. In Dentistry, cyanoacrylates have been used for suturing, pulp capping, as retrofilling material in endodontic surgeries, and as cervical plug for pulpless teeth bleaching. The biocompatibility of these adhesives has been the topic of many researches and subcutaneous implantation is an effective methodology for these studies. The present study evaluated the biocompatibility of three different cyanoacrylate-based adhesives. Thirty-six Wistar rats were used, divided into four groups of 9 animals each: A (control)--distilled water, B- cyanoacrylate ester (Super Bonder), C - n-butyl-cyanoacrylate (Histoacryl) and D- alpha-cyanoacrylate (Three Bond). The materials were dispensed in sponges of polyvinyl chloride, the animals were incised and the sponges were inserted in the subcutaneous tissue and sutured. Each group was sub-divided according to the time of sacrifice of the animals: 7, 21 and 45 days. Subjective analysis of the histologic material showed that all groups presented some degree of irritability, but the inflammatory reaction decreased with the experimental time in all groups. Group D showed an inflammatory reaction which was closer to that of the control group and was considered to have good biocompatibility. Groups B and C were similar and presented more aggressive inflammatory reactions when compared to the control group. Based on the results, it was concluded that alpha-cyanoacrylate (Three Bond) was the most biocompatible adhesive because it caused the lowest levels of inflammation. PMID- 18425245 TI - Facial dimensions, bite force and masticatory muscle thickness in preschool children with functional posterior crossbite. AB - Posterior crossbite may affect craniofacial growth and development. Thus, this study aimed to associate facial dimensions (by standardized frontal photographs) to masseter and anterior portion of the temporal muscle thickness (by ultrasonography) and maximal bilateral bite force in 49 children with deciduous and early mixed dentitions. They were distributed in four groups: deciduous normal occlusion (DNO, n = 15), deciduous-crossbite (DCB, n = 10), mixed-normal occlusion (MNO, n = 13) and mixed-crossbite (MCB, n = 11). Anterior facial height (AFH), bizygomatic width (FWB), and intergonial width (FWI) were determined and associated with muscle thickness and bite force, applying Pearsons coefficients and multiple logistic regression, with age, gender, body weight and height as the covariates. FWB and FWI were correlated positively with the masseter thickness, whereas AFH/FWB and AFH/FWI ratios had negative correlation, except in the DNO group. The correlation between AFH/FWB and bite force in the MCB group was significantly negative. A higher AFH/FWB in MNO and MCB led to a significantly higher probability for functional crossbite development. In the studied sample, it was observed that children in the early mixed dentition with a long-face trend showed lower bite force and higher probability to present functional posterior crossbite, without significant influence of the covariates. PMID- 18425246 TI - Streptococcus mutans counts in plaque adjacent to orthodontic brackets bonded with resin-modified glass ionomer cement or resin-based composite. AB - This study investigated the number of Streptococcus mutans CFU (colony forming units) in the saliva and plaque adjacent to orthodontic brackets bonded with a glass ionomer cement--GIC (Fuji Ortho) or a resin-based composite--RC (Concise). Twenty male and female patients, aged 12 to 20 years, participated in the study. Saliva was collected before and after placement of appliances. Plaque was collected from areas adjacent to brackets and saliva was again collected on the 15th, 30th, and 45th day after placement. On the 30th day, 0.4% stannous fluoride gel was applied for 4 minutes. No significant modification in the number of Streptococcus mutans CFU in saliva was observed after placement of the fixed orthodontic appliances. On the 15th day, the percentage of Streptococcus mutans CFU in plaque was statistically lower in sites adjacent to GIC-bonded brackets (mean = 0.365) than in those adjacent to RC-bonded brackets (mean = 0.935). No evidence was found of a contribution of GIC to the reduction of CFU in plaque after the 15th day. Topical application of stannous fluoride gel on the 30th day reduced the number of CFU in saliva, but not in plaque. This study suggests that the antimicrobial activity of GIC occurs only in the initial phase and is not responsible for a long-term anticariogenic property. PMID- 18425247 TI - Evaluation of mandibular range of motion in Brazilian children and its correlation to age, height, weight, and gender. AB - This study aimed to measure the active mandibular range of motion (ROM) (mouth opening: MO; right and left lateral movements: RL and LL; protrusion: P) in Brazilian children of both genders and of various ages, weights, and heights, and to establish correlations among such variables. Study subjects (n = 303) were healthy boys and girls, with ages ranging between 6 and 14 years, who were regular students of a public school in the state of Sao Paulo. Analysis of variance and intra-class correlation coefficients were considered significant for p < 0.05. Weak significant correlations were observed between mandibular ROM and age, height, and weight. No statistically significant differences (p > 0.05) were observed regarding mandibular ROM between gender groups. Mean mandibular ROM values showed significant increases (p < 0.05) in relation to age, height, and weight, except for RL (p > 0.05), in the studied age range. Significantly smaller (p < 0.05) mean mandibular ROM values were observed for the intervals of 6 and 7 years of age, 1.15-1.35 m, and 17.30-26.50 kg, in relation to the other ranges. Nonetheless, no differences were observed among mean mandibular ROM values in the ranges 8 to 12-14 years, 1.36-1.75 m, and 47-85 kg. Thus, it is suggested that weight, height, and age variables be considered when obtaining mandibular ROM values, particularly in children aged 6 to 7 years, measuring 1.15-1.35 m, and weighing between 17.3-46.5 kg. PMID- 18425248 TI - The influence of residual salivary fluoride from dentifrice on enamel erosion: an in situ study. AB - The objective of this study was to assess the salivary residual effect of fluoride dentifrice on human enamel subjected to an erosive challenge. This crossover in situ study was performed in two phases (A and B), involving ten volunteers. In each phase, they wore acrylic palatal appliances, each containing 3 human enamel blocks, during 7 days. The blocks were subjected to erosion by immersion of the appliances in a cola drink for 5 minutes, 4 times a day. Dentifrice was used to brush the volunteers teeth, 4 times a day, during 1 minute, before the appliance was replaced into the mouth. In phases A and B the dentifrices used had the same formulation, except for the absence (PD) or presence (FD) of fluoride, respectively. Enamel alterations were determined using profilometry, microhardness (%SMHC), acid- and alkali-soluble F analysis. The data were tested using ANOVA (p < 0.05). The concentrations (mean +/- SD) of alkali- and acid-soluble F (microgF/cm(2)) were, respectively, PD: 1.27 feminine +/- 0.70/2.24A +/- 0.36 and FD: 1.49 feminine +/- 0.44/2.24A +/- 0.67 (p > 0.05). The mean wear values (+/- SD, microm) were PD: 3.63 feminine +/- 1.54 and FD: 3.54 feminine +/- 0.90 (p > 0.05). The mean %SMHC values (+/- SD) were PD: 89.63 feminine +/- 4.73 and FD: 87.28 feminine +/- 4.01 (p > 0.05). Thus, we concluded that the residual fluoride from the fluoride-containing dentifrice did not protect enamel against erosion. PMID- 18425249 TI - Periodontal condition in patients with rheumatoid arthritis. AB - The purpose of this clinical study was to investigate if periodontal disease and rheumatoid arthritis (RA) are associated. The study included 39 RA patients (test group) and 22 age- and gender-matched healthy individuals (control group). Questionnaires on general and oral health were applied and a complete periodontal exam, including visible plaque, marginal bleeding, attachment loss (AL) and number of teeth present, was also performed by a single calibrated examiner. Diabetes mellitus patients and smokers were excluded. RA patients had fewer teeth, higher prevalence of sites presenting dental plaque and a higher frequency of sites with advanced attachment loss. Although the prevalence of dental plaque was higher in the test group (Chi-square test, p = 0.0006), the percentage of sites showing gingival bleeding was not different (Fishers exact test, p > 0.05). Based on our results, we suggest that there is an association between periodontal disease and RA. PMID- 18425250 TI - Image quality in partially erased DenOptix storage phosphor plates. AB - This study aimed at investigating the effect of the partial erasing of DenOptix system storage phosphor plates on the image quality of digital radiographs. Standardized digital radiographs were acquired of a phantom mandible, using size 2 intraoral DenOptix storage phosphor plates (n = 10). Subsequently, the active areas of the plates were placed in a viewing box with a constant light intensity of 1,700 lux for 130 seconds to achieve complete erasing (control plate), as well as for 0, 5, 10, 15, 20, 25, 34, 66, and 98 seconds, to compose the experimental group of partially erased plates. The same exposure settings were repeated using the control and experimental plates, which were scanned at a resolution of 300 dpi. Five radiologists independently examined the pairs of digital radiographs obtained with the control and partially erased plates, in random order, and indicated the best image for oral diagnosis. Cochran-Mantel-Haenszels chi-square test, at a significance level of 5%, was used to compare the percentages of superior quality images in each combination of control and partially erased plates, subjectively assessed. No significant differences were found between radiographic images acquired with control and partially erased plates, except for the combination of 0 second (30%) versus 130 seconds (70%), p = 0.0047. It can be concluded that, under adequate light intensity conditions, erasing intraoral DenOptix storage phosphor plates may require time intervals of as little as 5 seconds. PMID- 18425251 TI - Flexural and diametral tensile strength of composite resins. AB - This study evaluated the flexural strength (sf) and the diametral tensile strength (st) of light-cured composite resins, testing the hypothesis that there is a positive relation between these properties. Twenty specimens were fabricated for each material (Filtek Z250- 3M-Espe; AM- Amelogen, Ultradent; VE- Vit-l escence, Ultradent; EX- Esthet-X, Dentsply/Caulk), following ISO 4049 and ANSI/ADA 27 specifications and the manufacturers instructions. For the st test, cylindrical shaped (4 mm x 6 mm) specimens (n = 10) were placed with their long axes perpendicular to the applied compressive load at a crosshead speed of 1.0 mm/min. The sf was measured using the 3-point bending test, in which bar shaped specimens (n = 10) were tested at a crosshead speed of 0.5 mm/min. Both tests were performed in a universal testing machine (EMIC 2000) recording the fracture load (N). Strength values (MPa) were calculated and statistically analyzed by ANOVA and Tukey (a = 0.05). The mean and standard deviation values (MPa) were Z250-45.06 +/- 5.7; AM-35.61 +/- 5.4; VE-34.45 +/- 7.8; and EX-42.87 +/- 6.6 for st; and Z250-126.52 +/- 3.3; AM-87.75 +/- 3.8; VE-104.66 +/- 4.4; and EX-119.48 +/- 2.1 for sf. EX and Z250 showed higher st and sf values than the other materials evaluated (p < 0.05), which followed a decreasing trend of mean values. The results confirmed the study hypothesis, showing a positive relation between the material properties examined. PMID- 18425252 TI - Spectrophotometric assessment of the effects of 10% carbamide peroxide on enamel translucency. AB - Tooth shade results from the interaction between enamel color, enamel translucency and dentine color. A change in any of these parameters will change a tooths color. The objective of this study was to evaluate the changes occurring in enamel translucency during a tooth whitening process. Fourteen human tooth enamel fragments, with a mean thickness of 0.96 mm (+/- 0.3 mm), were subjected to a bleaching agent (10% carbamide peroxide) 8 hours per day for 28 days. The enamel fragment translucency was measured by a computer controlled spectrophotometer before and after the bleaching agent applications in accordance with ANSI Z80.3-1986--American National Standard for Ophthalmics--nonprescription sunglasses and fashion eyewear-requirements. The measurements were statistically compared by the Mann-Whitney non-parametric test. A decrease was observed in the translucency of all specimens and, consequently, there was a decrease in transmittance values for all samples. It was observed that the bleaching procedure significantly changes the enamel translucency, making it more opaque. PMID- 18425253 TI - Educational camp for children with asthma. PMID- 18425254 TI - Educational camp for children with asthma. AB - OBJECTIVE: To evaluate the impact of a 5-day educational camp program for children with asthma in terms of improving their knowledge of asthma and enhancing their performance in the use of inhaled medication and in physical activities. METHODS: Every day, the children received 20-min interactive educational sessions, the technique for using the metered-dose inhaler was reviewed, two peak flow readings were recorded, and the children performed physical activities that included breathing and relaxation exercises. A questionnaire regarding knowledge of asthma, as well as asthma triggers, asthma medications, misconceptions regarding asthma, and the use of spacers, was administered before and after the intervention. Correct use of inhaled medication and exercise-related symptoms were also evaluated before and after the intervention. RESULTS: A total of 37 children with asthma, aged 8-10 years (15 females and 22 males), were included in this study. Of those, 25% showed an improvement in the level of knowledge of asthma after the educational camp program, as evidenced by the greater number of correct answers on three of the twelve questions analyzed (p < 0.05). The exercise-related dyspnea scores decreased significantly (p < 0.05). The ability to use inhaled medication correctly was significantly improved after the intervention (p < 0.05). CONCLUSIONS: The asthma educational camp program can improve knowledge about specific questions, encourage participation in physical activities, and improve the asthma management skills of children. PMID- 18425255 TI - Prevalence of emotional and behavioral disorders in adolescents with asthma. AB - OBJECTIVE: To determine the prevalence of emotional and behavioral disorders in adolescents with asthma and to compare it with that of adolescents without asthma. METHODS: A transversal study using the Strengths and Difficulties Questionnaire, administered to adolescents with or without asthma, ranging from 14 to 16 years of age and randomly selected from schools in the city of Belo Horizonte, Brazil. RESULTS: The prevalence of emotional and behavioral disorders in adolescents with and without asthma was 20.4% (95% CI: 14.5-27.8%) and 9.0% (95% CI: 6.1-12.8%), respectively. Among adolescents with asthma, 56.6% (95% CI: 48.3-64.5%) presented normal scores, and 23.0% (95% CI 16.8-30.7%) presented borderline scores. Among adolescents without asthma, 75.0% (95% CI: 69.7-79.6%) presented normal scores, and 16.0% (95% CI: 12.2-20.7%) presented borderline scores. The median total score on the questionnaire was 14 and 12 among subjects with and without asthma, respectively (p < 0.01). In the final logistic regression model, adjusted for socioeconomic variables, the association between emotional/behavioral disorders and the following variables remained significant: being female (OR = 1.98; 95% CI: 1.10-3.56; p = 0.02) and having asthma (OR = 2.66; 95% CI: 1.52-4.64; p = 0.001). CONCLUSIONS: The prevalence of emotional and behavioral disorders is higher in adolescents with asthma than in those without asthma, underscoring the need for a holistic, interdisciplinary approach. PMID- 18425256 TI - Thoracoscopy in the treatment of pleural empyema in pediatric patients. AB - OBJECTIVE: To evaluate the results of thoracoscopy for the treatment of pleural empyema in pediatric patients. METHODS: A retrospective study of 117 patients who underwent mediastinoscopy or video-assisted thoracoscopy for pleural empyema treatment. General anesthesia and single-lumen oral intubation were used. Surgery was indicated when there was pleural effusion and no clinical and radiological response to clinical treatment (antibiotics, physiotherapy and thoracocentesis) or severe sepsis, together with loculated pleural effusion (confirmed through ultrasound or computed tomography of the chest). RESULTS: Between February of 1983 and July of 2006, 117 thoracoscopies were performed in patients ranging in age from 5 months to 17 years (mean, 4 years). Mean time for thoracic drainage was 9 days (range, 2-33 days), and mean period of hospitalization was 16.4 days (range, 4 to 49 days). One patient (0.8%) died after surgery, and persistent fistula was observed in 33 patients (28%). In 7 cases (6%), open thoracotomy with pulmonary decortication was performed due to the disposition of the empyema. CONCLUSIONS: Management of pleural empyema in this age bracket is still controversial, and surgical indication is often delayed, particularly when there are multiple loculations or severe sepsis. Early thoracoscopy yields a better clinical outcome for pediatric patients with pleural empyema, with apparent decreased morbidity and mortality, earlier chest tube removal, earlier hospital discharge and improved response to antibiotic therapy. PMID- 18425257 TI - Oxygen outflow delivered by manually operated self-inflating resuscitation bags in patients breathing spontaneously. AB - OBJECTIVE: To determine the oxygen outflow delivered by seven different models of manually operated self-inflating resuscitation bags (with and without an oxygen reservoir connected), which were tested using different oxygen supply rates without manipulating the bag, by simulating their use in patients breathing spontaneously. METHODS: The oxygen outflow was measured using a wall oxygen flow meter and a flow meter/respirometer attached to the bag, together with another flow meter/respirometer attached to the patient connection port. The resuscitation bags that allow the connection of an oxygen reservoir were tested with and without this device. All resuscitation bags were tested using oxygen supply rates of 1, 5, 10, and 15 L/min. Statistical analyses were performed using analysis of variance and t-tests. RESULTS: The resuscitation bags that allow the connection of an oxygen reservoir presented a greater oxygen outflow when this device was connected. All resuscitation bags delivered a greater oxygen outflow when receiving oxygen at a rate of 15 L/min. However, not all models delivered a sufficient oxygen outflow even when the two previous conditions were satisfied. CONCLUSIONS: Of the resuscitation bags studied, those that allow the connection of an oxygen reservoir must have this reservoir connected to the bag when used as a source of oxygen in nonintubated spontaneously breathing patients. All of the models studied should receive oxygen at a rate > 15 L/min. It is not safe to use manually operated self-inflating resuscitation bags for this purpose without knowing their characteristics. PMID- 18425258 TI - Evaluation of adenosine deaminase in the diagnosis of pleural tuberculosis: a Brazilian meta-analysis. AB - OBJECTIVE: To evaluate Brazilian studies by summarizing the accuracy of adenosine deaminase in the diagnosis of pleural tuberculosis, with the objective of lending support to the movement to make the test part of the routine investigation of pleural effusions. METHODS: A search for Brazilian studies related to the determination of adenosine deaminase levels in the pleural liquid was carried out. These studies were evaluated and included in this study. The data were analyzed using summary receiver operating characteristic (SROC) curves, which enabled the studies to be collected and evaluated regarding the accuracy of the diagnosis. As for the global values of sensitivity and specificity, the Bayes' theorem was applied to calculate the post-test probabilities in different prevalences of the disease. RESULTS: Twenty-five studies dating from 1987 to 2005 and including enough information to be used in the meta-analysis were identified. After evaluation, nine studies were included, totaling 1674 patients. According to the SROC curve, a sensitivity of 91.8% (95% CI: 89.8-93.6%) and a specificity of 88.4% (95% CI: 86.0-90.5%) were found, with an area of 0.969 below the curve. The overall odds ratio was 112.0 (95% CI: 51.6-243.2). Considering a prevalence of tuberculosis of 50% (considered neutral), the post-test probability was 88.7% for a positive test and 91.5% for a negative test. CONCLUSIONS: Despite the differences found among studies, it is possible to conclude that the determination of adenosine deaminase levels has high accuracy in the diagnosis of the pleural tuberculosis and should be used as a routine test in its investigation. PMID- 18425259 TI - Completeness of tuberculosis control program records in the case registry database of the state of Espirito Santo, Brazil: analysis of the 2001-2005 period. AB - OBJECTIVE: To investigate the completeness of tuberculosis control program records for the 2001-2005 period in the case registry database of the state of Espirito Santo, Brazil. METHODS: A descriptive study using secondary data analysis. The following variables were studied: race/color; educational level, supervised treatment, sputum culture at treatment outcome, HIV status, smear sputum microscopy at the sixth month of treatment and treatment outcome. RESULTS: Using technical parameters of the case registry database of the Brazilian National Ministry of Health regarding the percentage of completeness of data on the reporting forms, the tuberculosis database of the state of Espirito Santo can be classified as fair to excellent. The state of Espirito Santo has 78 cities, 8 of which are considered priorities for tuberculosis control. For the variables studied, the cities not considered priorities presented better completeness of records than did those considered priorities, the exception being the variable treatment outcome, for which the percentage of completeness was comparable among all of the cities. CONCLUSIONS: Since the cities that are considered priorities accounted for 70.2% of the total number of reports, the responsible officials in those cities should improve the quality of those reports in terms of the completeness of forms and the updating of data, which are fundamental to the reliability of epidemiological analyses. That will make it possible to plan activities aimed at the prevention and control of tuberculosis in the state. PMID- 18425260 TI - Allergic rhinitis: epidemiological aspects, diagnosis and treatment. AB - This study was a review of the literature on the epidemiological, clinical, diagnostic and therapeutic aspects of allergic rhinitis. Bibliographic searches were based on the information contained within the Medline, Latin American and Caribbean Health Sciences Literature and HighWire databases, covering the last thirty years and using the following search terms: 'allergic rhinitis', 'epidemiology', 'diagnosis' and 'treatment'. Sixty articles were selected. This study describes the increase in the prevalence of allergic rhinitis, its relationship with asthma, the diagnostic criteria and the treatment. The classification of allergic rhinitis and strategies for its treatment are presented. Therapeutic modalities presented and discussed include the administration of antihistamines, corticosteroids, immunotherapy, anti leukotrienes, sodium cromoglycate and anti-IgE antibodies, as well as minimizing exposure to inhaled allergens. Finally, the importance of the management of allergic rhinitis in public health is emphasized. PMID- 18425261 TI - Video-assisted thoracoscopic removal of foreign bodies from the pleural cavity. AB - Video-assisted thoracoscopy is a minimally invasive surgical technique. It is the procedure of choice for the treatment of certain lung and pleural diseases. It has been increasingly used in the acute phase of thoracic trauma. Another indication for its use, which is rarely described in the literature, is for the removal of retained intrathoracic foreign bodies. We report the cases of two hemodynamically stable gunshot victims in whom the bullet was removed from the pleural cavity through video-assisted thoracoscopy, thereby avoiding the need for thoracotomy. PMID- 18425262 TI - Treatment of pulmonary actinomycosis with levofloxacin. AB - Actinomycosis is a chronic suppurative bacterial infection characterized by multiple abscesses, fistulous pathways, and fibrosis involving the face, neck, chest, and abdomen. It is caused by an anaerobic Gram-positive saprophytic bacterium (Actinomyces). Primary actinomycosis of the lung is a rare disease that probably results from aspiration of oropharyngeal secretions. It can present as a chronic respiratory disease. The treatment of choice is antibiotic therapy with penicillin. The authors report the case of a 55-year-old female diagnosed with pulmonary actinomycosis and successfully treated with levofloxacin. PMID- 18425264 TI - Speciation of antimony (III) and antimony (V) using hydride generation for meglumine antimoniate pharmaceutical formulations quality control. AB - The pentavalent antimonies, mainly the meglumine antimoniate, are recommends as first-choice medicines for leishmaniasis therapy. In this work we described the development of formulations of meglumine antimoniate injectable medication, as well as the analytical methodology used in the selective determination of Sb(III) and Sb(Total) by hydride generation - inductively coupled plasma atomic emission spectrometry (HG-ICP-AES) and ICP-AES, respectively. On that purpose the analytical methodology was developed focusing on the HG-ICP-AES technique. The formulations using propylene glycol/water as vehicles in a 20:80 proportion were more appropriate for subsequent use in industrial scale. These formulations also showed a lower variation on Sb(III) percentage, no need of buffer solution to stabilize the formulation and no influence of the autoclaving in the quality of the product. The results of the development of the analytical methodology point out the proposed method as an efficient alternative for the determination of Sb(III) in the presence of large quantities of Sb(V) in injectable solutions of meglumine antimoniate, in a selective, linear, accurate and precise manner. In addition, the method showed a low limit of quantification, less interference of the matrix, and more resilience than batch techniques proposed in the Brazilian Pharmacopeia. PMID- 18425263 TI - The hope for an HIV vaccine based on induction of CD8+ T lymphocytes--a review. AB - The only long-term and cost-effective solution to the human immunodeficiency virus (HIV) epidemic in the developing world is a vaccine that prevents individuals from becoming infected or, once infected, from passing the virus on to others. There is currently little hope for an AIDS vaccine. Conventional attempts to induce protective antibody and CD8(+) lymphocyte responses against HIV and simian immunodeficiency virus (SIV) have failed. The enormous diversity of the virus has only recently been appreciated by vaccinologists, and our assays to determine CD8(+) lymphocyte antiviral efficacy are inadequate. The central hypothesis of a CTL-based vaccine is that particularly effective CD8(+) lymphocytes directed against at least five epitopes that are derived from regions under functional and structural constraints will control replication of pathogenic SIV. This would be somewhat analogous to control of virus replication by triple drug therapy or neutralizing antibodies. PMID- 18425265 TI - The utility of the polymerase chain reaction assay for aetiologic definition of unspecified bacterial meningitis cases. AB - Most patients with acute suppurative meningitis are otherwise healthy individuals with regard to immune mechanisms against invasive bacterial disease. This medical emergency is among the most dramatic and potentially ravaging diseases that affect humans, particularly young children. The illness often strikes suddenly, and can either result in death or leave the survivors with significant neurological dysfunctions. The demonstration of a bacterial aetiology is necessary for decisions regarding treatment and prophylaxis. Conventional bacteriological methods frequently fail to identify an agent, as a result of administration of antibiotics or delayed lumbar punctures. We investigated the major aetiologic sources of unspecified bacterial meningitis cases (G00.9, ISCD 10) by polymerase chain reaction (PCR)-based identification of Neisseria meningitidis (crgA), Streptococcus pneumoniae (ply) and Haemophilus influenzae (bexA) in cerebrospinal fluid samples. The multiplex PCR detected N. meningitidis in 92%, S. pneumoniae in 4% and H. influenzae in 1% of the 192 clinical samples assayed; 3% were negative for all three DNA targets. Bacterial DNA detection was found to be a valuable adjunct to enhance bacterial meningitis surveillance when the yield of specimens by culture is reduced. The implementation of PCR assays as a diagnostic procedure in Public Health Laboratories is perceived to be a significant advance in the investigation of bacterial meningitis. PMID- 18425266 TI - Prevalence of HIV type 1 drug resistance mutations in treatment-naive and experienced patients from resource-limited settings with universal access to antiretroviral therapy: a survey in two small Brazilian cities. AB - Concerns have been raised that universal availability of antiretroviral agents in resource-limited settings might lead to the emergence and spread of resistant strains. We present the largest survey on human immunodeficiency virus type 1 (HIV-1) resistance among treatment-naive and experienced patients followed in small, relatively underprivileged cities in Brazil with universal availability to standard of care antiretroviral combinations. Samples were collected between 2004 and 2006 from 95 patients followed in the cities of Saquarema and Santo Antonio de Padua, state of Rio de Janeiro. A proviral fragment encompassing protease and reverse transcriptase (RT) regions was generated and drug susceptibility level was inferred. Among 50 strains from drug-naive subjects, one (2%) had intermediate-level resistance to RT inhibitors. Among 38 patients on therapy as of sampling, 28 (73.7%) had plasma viral load (PVL) below detection limit (26 of whom without evidence of resistance mutations) and 11 (28.9%) harbored strains with reduced susceptibility. Only two strains harbored both protease and RT inhibitor mutations. Among seven patients who were off-treatment as of sampling, two (28.5%) harbored strains with reduced susceptibility to RT inhibitors. The relatively high frequency of undetectable PVL among patients on treatment and the overall low prevalence of resistance-associated mutations are reassuring. Continued surveillance, however, is necessary. PMID- 18425267 TI - Molecular and morphological characterization of heterorhabditid entomopathogenic nematodes from the tropical rainforest in Brazil. AB - Despite massive losses of primary forest, the Amazonian rainforest remains an extremely rich source of biodiversity. In recent years, entomopathogenic nematodes (EPNs) have been isolated from soil in various parts of the world and used successfully as biological control agents against numerous insect pests. Therefore, a sampling in the rainforest of Monte Negro, Rondonia, Brazil was conducted with the aim of discovering new strains and/or species of EPNs for future development as biological control agents. From 156 soil samples taken at nine collecting sites, 19 isolates were obtained, all of them belonging to the genus Heterorhabditis. Four strains were subjected to detailed morphological and molecular evaluation. Based on morphometrics and internal transcribed spacer (ITS) sequence data, the strains LPP1, LPP2 and LPP4 were identified as Heterorhabditis indica, whereas LPP7 was considered Heterorhabditis baujardi. Comparative analysis of the ITS1 sequence of H. indica and H. baujardi isolates showed a polymorphic site for the restriction enzyme Tth 111 that could be used to distinguish the two species. Consequently, strains LPP1, LPP2, LPP3, LPP4, and LPP9 were identified as H. indica, whereas LPP5, LPP7, LPP8 and LPP10 were identified as H. baujardi. PMID- 18425268 TI - Chromosome variability in the Chagas disease vector Rhodnius pallescens (Hemiptera, Reduviidae, Rhodniini). AB - Rhodnius pallescens is the main vector of Trypanosoma cruzi in Panama and one of the most relevant secondary vectors in Colombia. Despite the importance of this species, there is limited knowledge about the genetic variability along its geographical distribution. In order to evaluate the degree of karyotype variability we analyzed the meiotic behavior and banding pattern of the chromosomes of 112 males of R. pallescens coming from different regions of Colombia and Panama. Using the C-banding technique we identified two chromosomal patterns or cytotypes characterized by differences in the amount, size and distribution of constitutive heterochromatic regions in the chromosome complement (2n = 20 autosomes plus XY in males). The individuals can be easily classified in each cytotype by the analysis of the chromosomes during first meiotic prophase. The frequencies of the cytotypes are variable according to the geographic origin of the populations. This chromosomal divergence together with morphological data supports the existence of three genetically different populations of R. pallescens and provides new information to understand the distribution dynamics of this species. PMID- 18425269 TI - Preliminary characterization of a Rhodnius prolixus hemolymph trypanolytic protein, this being a determinant of Trypanosoma rangeli KP1(+) and KP1(-) subpopulations' vectorial ability. AB - Rhodnius prolixus is the main Trypanosoma rangeli vector in several Latin American countries and is susceptible to infection with KP1(+) strains; however, it presents an invasion-resistant response to KP1(-) strains. The present work has identified a trypanolytic protein against T. rangeli KP1(-) in the R. prolixus hemolymph which was fractioned with ammonium sulfate (following dialysis). The results revealed a protein component which did not depend on divalent cations for its biological function whilst keeping its trypanolytic activity at temperatures ranging from -20 masculineC to 37 masculineC, at 7.0 to 10.5 pH. The protein was partially purified by gel filtration chromatography and ionic exchange chromatography. The major component presented a molecular weight of around 79 kDa and an isoelectric point between 4.9 and 6.3 and may be directly related to hemolymph trypanolytic activity against T. rangeli KP1(-) populations. PMID- 18425270 TI - The epidemiology and antigenic characterization of influenza viruses isolated in Curitiba, South Brazil. AB - Several studies conducted all over the world have reported that the influenza virus is associated with great morbidity and mortality rates. In this study, we analyzed the incidence of the influenza virus between 2000 and 2003 in Curitiba. We studied 1621 samples obtained from outpatients and hospitalized patients of both sexes and all ages. The study was conducted at the local primary care health units (outpatients) and at the tertiary care unit (hospitalized) of the General Hospital of the Federal University in the state of Parana, Brazil. Nasopharyngeal aspirates and, eventually, bronchoalveolar lavage were assayed for the presence of viral antigens, either by indirect immunofluorescence or cell culture. Of the samples studied, 135 (8.3%) were positive for influenza virus, and of those, 103 (76.3%) were positive for type A and 32 (23.7%) for type B. Additionally, positive samples were analyzed by reverse transcription followed by polymerase chain reaction and subtypes H1 and H3 were identified from this group. A high incidence of positive samples was observed mainly in the months with lower temperatures. Furthermore, outpatients showed a higher incidence of influenza viruses than hospitalized patients. PMID- 18425271 TI - IgG and IgG2 antibodies from cattle naturally infected with Anaplasma marginale recognize the recombinant vaccine candidate antigens VirB9, VirB10, and elongation factor-Tu. AB - Anaplasma marginale is an important vector-borne rickettsia of ruminants in tropical and subtropical regions of the world. Immunization with purified outer membranes of this organism induces protection against acute anaplasmosis. Previous studies, with proteomic and genomic approach identified 21 proteins within the outer membrane immunogen in addition to previously characterized major surface protein1a-5 (MSP1a-5). Among the newly described proteins were VirB9, VirB10, and elongation factor-Tu (EF-Tu). VirB9, VirB10 are considered part of the type IV secretion system (TFSS), which mediates secretion or cell-to-cell transfer of macromolecules, proteins, or DNA-protein complexes in Gram-negative bacteria. EF-Tu can be located in the bacterial surface, mediating bacterial attachment to host cells, or in the bacterial cytoplasm for protein synthesis. However, the roles of VirB9, VirB10, and TFSS in A. marginale have not been defined. VirB9, VirB10, and EF-Tu have not been explored as vaccine antigens. In this study, we demonstrate that sera of cattle infected with A. marginale, with homologous or heterologous isolates recognize recombinant VirB9, VirB10, and EF Tu. IgG2 from naturally infected cattle also reacts with these proteins. Recognition of epitopes by total IgG and by IgG2 from infected cattle with A. marginale support the inclusion of these proteins in recombinant vaccines against this rickettsia. PMID- 18425272 TI - Molecular identification of Rickettsia felis in ticks and fleas from an endemic area for Brazilian Spotted Fever. AB - Rickettsioses are arthropod-borne diseases caused by parasites from the Order Rickettsiales. The most prevalent rickettsial disease in Brazil is Brazilian Spotted Fever (BSF). This work intends the molecular detection of those agents in ectoparasites from an endemic area of BSF in the state of Espirito Santo. A total of 502 ectoparasites, among them Amblyomma cajennense, Amblyomma dubitatum (A. cooperi), Riphicephalus sanguineus, Anocentor nitens and Ctenocephalides felis, was collected from domestic animals and the environment and separated in 152 lots according to the origin. Rickettsia sp. was detected in pools of all collected species by amplification of 17 kDa protein-encoding gene fragments. The products of PCR amplification of three samples were sequenced, and Rickettsia felis was identified in R. sanguineus and C. felis. These results confirm the presence of Rickettsia felis in areas previously known as endemic for BSF, disease caused by Rickettsia rickettsii. Moreover, they show the needing of further studies for deeper knowledge of R. felis-spotted fever epidemiology and differentiation of these diseases in Brazil. PMID- 18425273 TI - Adenoviruses C in non-hospitalized Mexican children older than five years of age with acute respiratory infection. AB - Adenoviruses (AdV) are commonly involved in acute respiratory infections (ARI), which cause high morbidity and mortality in children. AdV are grouped in six species (A-F), which are associated with a wide range of diseases. The aim of this study was to identify the AdV species infecting non-hospitalized Mexican children with ARI symptoms, attending to the same school. For that, a PCR/RFLP assay was designed for a region of the hexon gene, which was chosen, based on the bioinformatical analysis of AdV genomes obtained from GenBank. A total of 100 children's nasopharyngeal samples were collected from January to June, 2005, and used for viral isolation in A549 cells and PCR/RFLP analysis. Only 15 samples produced cytopathic effect, and in all of them AdV C was identified. AdV C was also identified in eight additional nasopharyngeal samples which were negative for viral isolation. In summary, this outpatient population showed a rate of AdV infection of 23%, and only AdV C was detected. PMID- 18425275 TI - First record in South America of Didymosulcus palati and Didymosulcus philobranchiarca (Digenea, Didymozoidae) with new hosts records and pathological alterations. AB - Two species of Didymozoidae, Didymosulcus palati (Yamaguti 1970) and Didymosulcus philobranchiarca (Yamaguti 1970) were reported for the first time in South America, Atlantic Ocean, parasitizing three different tuna species from the coast of Rio de Janeiro, Brazil: Thunnus atlanticus (Lesson), Thunnus albacares (Bonnaterre) and Thunnus obesus (Lowe). Pairs of D. philobranchiarca were found on gill arches of T. albacares and T. obesus, in longitudinal rows of yellow cysts located inside grooves in the hard denticle palate (new site) of the three hosts species studied, and as disperse groups of cysts in the operculum (new site) and gill arches of T. atlanticus (new host record). D. palati occurred as disperse groups of encysted worm pairs in the gill arches of T. albacares and T. obesus and in gill arches and operculum of T. atlanticus (new host record). The pathological alterations induced by D. philobranchiarca in the palate of T. atlanticus are described for the first time. Original measurements and figures are presented. PMID- 18425274 TI - Hospitalization due to norovirus and genotypes of rotavirus in pediatric patients, state of Espirito Santo. AB - Viruses are the leading cause for hospitalization due to gastroenteritis worldwide. Group A rotaviruses (RV) are the most prevalent and are assorted in glycoproteins (G) and protease sensitive (P) dual genotypes based on polymorphic genes that encode the external VP7 and VP4 capsid proteins, respectively. Noroviruses (NoV) have increasingly answered by sporadic gastroenteritis. This study aimed to determine the prevalence of NoV and RV in 68 hospitalized children, between July 2004 and November 2006, at a pediatric hospital in Vitoria city, state of Espirito Santo, Southeastern Brazil. Nucleic acid was extracted from fecal suspension following the guanidine-silica procedure. Reverse transcriptase-polymerase chain reaction (RT-PCR) and polyacrylamide gel electrophoresis were employed for NoV and RV detection, respectively. RV genotyping was accomplished using RT-PCR followed by heminested multiplex PCR with specific primers for the most prevalent types of G and P. Fecal samples were positive for NoV and RV in 39.7% (27/68) and 20.5% (14/68), respectively and together were responsible for 60% (41/68) of the cases. RV genotypes were: 50% G9P[8], 28.7% G2P[4], 7.1% G1P[8], G2P[8] and G?P[8]. Vomit was a prominent manifestation observed in 92% and 85% of the NoV and RV cases, respectively. The median hospitalization was 5 and 5.5 days for the patients infected with NoV and RV, respectively. The data showed that NoV prevailed over RV and it also corroborated the emergence of RV G9 genotype followed by G2P[4], reinforcing the need for RV genotype surveillance. PMID- 18425276 TI - Diptera Brachycera found inside the esophagus of a mummified adult male from the early XIX century, Lisbon, Portugal. AB - Fly puparia and adult fragments of diptera muscid were found inside the esophagus of a mummified body from the early XIX century, buried inside the crypt of the Sacrament Church (Lisbon, Portugal). The identification of the material revealed a monospecific colonization by Ophyra capensis (Wiedemann) (Diptera: Muscidae), a species known to invade corpses in the ammoniacal fermentation wave. This species can be found in corpses kept indoors, not available to the early waves of blowflies (Diptera: Calliphoridae). In the present case, the number of pupae and their developmental stage suggest that the female invaded the mummified corpse through the partially opened mouth and the oviposition took place directly inside the esophagus. This is the first case of O. capensis infesting internal organs of an intact corpse. The use of chemical products for the embalming process probably explains why external colonization did not occur. PMID- 18425277 TI - Impact of moisture on survival of Aedes aegypti eggs and ovicidal activity of Metarhizium anisopliae under laboratory conditions. AB - The effect of relative humidity (43%, 75%, 86% and > 98%) on Aedes aegypti eggs treated with Metarhizium anisopliae or water only was tested for up to a six months exposure at 25 degrees C. Survival of larvae inside eggs was clearly affected by the lowest humidity (43%) tested, and eclosion diminished at all humidities after increasing periods of exposure. M. anisopliae showed to have a strong ovicidal activity only at humidity close to saturation. No difference of activity was found between conidia and hyphal bodies tested. This fungus affected larvae inside eggs and has potential as a control agent of this important vector in breeding sites with high moisture. PMID- 18425278 TI - Reproductive isolation between sympatric and allopatric Brazilian populations of Lutzomyia longipalpis s.l. (Diptera: Psychodidae). AB - Lutzomyia longipalpis s.l., the main vector of Leishmania chagasi in Latin America, is a species complex although the exact number of siblings is yet unknown. In Brazil, the siblings differ in male copulatory courtship songs and pheromones that most certainly act as pre-zygotic reproductive barriers. Here we analysed the reproductive isolation between three allopatric and two sympatric populations of Lu. longipalpis s.l. from Brazil. The results indicate a strong copulatory and pre-mating isolation between the three allopatric populations. In addition, the results also indicate a stronger pre-mating isolation between the two sympatric siblings than between the three allopatric ones, suggesting a role for reinforcement in the speciation of the Lu. longipalpis s.l. complex. PMID- 18425279 TI - Evidence-based healthcare for all: a new era? PMID- 18425280 TI - Risk factors associated with developmental abnormalities among high-risk children attended at a multidisciplinary clinic. AB - CONTEXT AND OBJECTIVE: Knowledge of risk factors associated with child development disorders is essential for delivering high-quality childcare. The objective here was to evaluate the relationships between risk factors and occurrences of developmental abnormalities among children attended at a reference clinic for children at risk of developmental abnormalities. DESIGN AND SETTING: Retrospective study at a multidisciplinary reference center, Embu, Sao Paulo. METHODS: All cases followed up for more than three months between 1995 and 2003 were reviewed. The risk factors assessed were low birth weight, gestational age, length of stay in neonatal ward, perinatal asphyxia, mothers age < 18 years, congenital infections, malformations and low mothers education level. Developmental abnormalities were defined according to developmental tests and assessments by the clinics professionals. The statistical analysis consisted of the chi-squared test for comparing categorical variables and a logistic regression model for multivariate analysis. RESULTS: 211 children were followed up for more than three months. Developmental abnormalities occurred in 111 (52.6%). Univariate analysis showed significant relationships between developmental abnormality and low birth weight, perinatal asphyxia, length of stay > 5 days, prematurity and mothers age 18 years and older. Low birth weight, history of perinatal asphyxia and mothers age continued to be significant in multivariate analysis. CONCLUSIONS: Special attention must be paid to the development of low birth weight infants and/or infants with histories of neonatal complications. Low birth weight is easily assessed and should be considered to be an important marker when defining guidelines for following up child development. PMID- 18425281 TI - Postoperative study of vital capacity and ventilation measurements following elective craniotomy. AB - CONTEXT AND OBJECTIVE: Changes in pulmonary function commonly occur after general surgery. The aims were to evaluate vital capacity, tidal volume and respiratory frequency among patients undergoing elective craniotomy and to determine possible correlations of these parameters with surgery duration and etiology for neurosurgery. DESIGN AND SETTING: Prospective, open study at a tertiary university hospital. METHODS: Twenty-six patients underwent elective craniotomy for aneurysm clipping (11) or tumor resection (15). Vital capacity (VC), tidal volume (TV), minute volume (VE) and respiratory rate were determined before the operation and on the first to fourth postoperative days. RESULTS: There were significant decreases of 25% in VC, 22% in TV and 12% in VE (p < 0.05) and no significant increase in respiratory frequency (5%) on the first postoperative day. VE returned to baseline on the second postoperative day and TV on the third postoperative day, while VC was 8% lower on the fourth postoperative day, compared with before the operation (p < 0.05). VC reduction was significantly greater in patients undergoing aneurysm clipping (43%) than in patients undergoing tumor resection (14%) when surgery duration was more than four hours (p < 0.05), with no significant change when surgery duration was less than four hours. CONCLUSION: Reductions in VC, TV and VE were observed during the postoperative period in patients undergoing aneurysm clipping or tumor resection. The reductions in VC and TV were greater in patients undergoing craniotomy due to aneurysm and with longer surgery duration. PMID- 18425282 TI - Recurrence of cervical intraepithelial neoplasia grades 2 or 3 in HIV-infected women treated by large loop excision of the transformation zone (LLETZ). AB - CONTEXT AND OBJECTIVE: Women infected by HIV are more likely to have cervical cancer and its precursors. Treatment of the precursor lesions can prevent this neoplasia. The aim of this study was to assess the likelihood of recurrent cervical intraepithelial neoplasia grades 2 or 3 (CIN 2-3) in HIV-infected women, compared with HIV-negative women, all treated by large loop excision of the transformation zone (LLETZ). DESIGN AND SETTING: A cohort study in Instituto Fernandes Figueira, Fundacao Oswaldo Cruz (IFF-Fiocruz), Rio de Janeiro. METHOD: 55 HIV-positive and 212 HIV-negative women were followed up after LLETZ for CIN 2 3 (range: 6-133 months). RESULTS: The incidence of recurrent CIN 2-3 was 30.06/10,000 woman-months in the HIV-positive group and 4.88/10,000 woman-months in the HIV-negative group (relative risk, RR = 6.16; 95% confidence interval, CI: 2.07-18.34). The likelihood of recurrence reached 26% at the 62nd month of follow up among the HIV-positive women, and remained stable at almost 0.6% at the 93rd month of follow-up among the HIV-negative women. We were unable to demonstrate other prognostic factors relating to CIN recurrence, but the use of highly active antiretroviral therapy (HAART) may decrease the risk of this occurrence among HIV patients. CONCLUSION: After LLETZ there is a higher risk of recurrence of CIN 2-3 among HIV-positive women than among HIV-negative women. This higher risk was not influenced by margin status or grade of cervical disease treated. The use of HAART may decrease the risk of this occurrence in HIV patients. PMID- 18425283 TI - Accuracy study on "Osteorisk": a new osteoporosis screening clinical tool for women over 50 years old. AB - CONTEXT AND OBJECTIVE: Osteoporosis is the greatest cause of quality-of-life reductions, morbidity and mortality among postmenopausal women, with growing incidence as populations age. Clinical tools like Osteorisk provide an easy access and low-cost alternative method that helps physicians to reduce the need for dual-energy X-ray absorptiometry (DXA), the expensive gold standard examination for diagnosing osteoporosis. The aim here was to study the accuracy of Osteorisk using heel ultrasonography for bone mineral density (BMD). DESIGN AND SETTING: Cross-sectional study, at Faculdade de Medicina do ABC. METHODS: A structured questionnaire was applied to 615 postmenopausal women, with anthropometric measurements, Osteorisk calculations and quantitative ultrasound on the heel using Sonost 2000 equipment. RESULTS: 461 women were included, with mean age 60 +/- 9 years, weight 67.6 +/- 12.9 kg and body mass index (BMI) 28.8 +/- 5.0 kg/m(2). Their Osteorisk classifications were: 61.0% low-risk, 28.4% medium-risk and 10.6% high-risk. Quantitative ultrasound showed 81.3% low-risk, 10.0% medium-risk and 8.7% high-risk regarding osteoporosis. Statistically significant results were observed (p < 0.001) when Osteorisk was correlated with age, years since menopause and BMI. Correlating these same variables with quantitative ultrasound, statistically significant results were observed for age (p < 0.001), years since menopause (p < 0.001) and BMI (p < 0.006). The sensitivity, specificity, negative predictive value and positive predictive value for Osteorisk were 64%, 6.7%, 89% and 30.6%, respectively. CONCLUSION: Osteorisk is a valid tool for screening for women at low risk of osteoporosis, making it possible for these women not to have to undergo densitometry. PMID- 18425284 TI - Relationship between stuttering severity in children and their mothers speaking rate. AB - CONTEXT AND OBJECTIVE: Stuttering is a complex disease that influences occupational, social, academic and emotional achievements. The aim of this study was to correlate the stuttering severity index with speaking rates of mothers and children. DESIGN AND SETTING: Cross-sectional study, at the child rehabilitation clinics of Tehran city. METHODS: 35 pairs of mothers and their children who stuttered were studied. There were 29 boys and six girls, of mean age 8.5 years (range: 5.1-12.0). Speech samples from the mother-child pairs were audiotaped for approximately 15 minutes, until a reciprocal verbal interaction had been obtained. This sample was then analyzed in accordance with a stuttering severity index test and speaking rate parameters. RESULTS: The research results outlined a significant relationship between the mothers speaking rate and their childrens stuttering severity. CONCLUSION: The results suggest that the mothers speaking rate should be incorporated in the assessment and treatment of stuttering. PMID- 18425285 TI - Divergences in antihypertensive therapy in special situations in nephrology. AB - CONTEXT AND OBJECTIVE: The choice of an antihypertensive drug is based on several criteria and specific situations give rise to doubt and controversy. The aim here was to evaluate physicians approaches towards treatment with antihypertensive agents in specific situations. DESIGN AND SETTING: Cross-sectional study, at Universidade Federal de Sao Paulo, Sao Paulo. METHODS: A questionnaire was applied during a nephrology meeting to evaluate individual approaches towards each hypothetical clinical situation. The questionnaire consisted of five multiple-choice questions (clinical cases) concerning controversial aspects of antihypertensive therapy. RESULTS: A total of 165 questionnaires were analyzed. Most participants were nephrologists (93.2%). There was a preference for angiotensin-converting enzyme (ACE) inhibitors in at least two of the cases. Only 57.2% of the physicians were correct in choosing beta-blockers as the first-line drugs for patients with ischemic coronary disease. Moreover, 66.2% chose ACE inhibitors as the first-line drugs for patients with chronic kidney disease and proteinuria. About 5% of the physicians did not follow the current recommendations for the use of ACE inhibitors in diabetic patients with microalbuminuria. The most controversial question concerned the first-line drug for advanced chronic kidney disease. Most physicians were correct in choosing calcium channel blockers and avoiding ACE inhibitors in renovascular hypertension in the case of a patient with a single functioning kidney. CONCLUSIONS: Most physicians adopted the correct approach, but some had an alternative strategy for the same situations that was not based on evidence. PMID- 18425286 TI - Cutaneous melanoma: descriptive epidemiological study. AB - CONTEXT AND OBJECTIVE: Cutaneous melanoma represents around 3% of all skin tumors. About 20% of such patients will have advanced disease and will die before reaching five years of survival. The aim of this paper was to describe the clinical and histopathological variables and their correlations. DESIGN AND SETTING: Retrospective, descriptive, epidemiological study at the Melanoma Unit, Dermatological Clinic, Irmandade da Santa Casa de Misericordia, Sao Paulo. METHODS: Records from 364 cases between May 1993 and January 2006 were analyzed. The frequencies of all study variables and their 95% confidence intervals were determined. The chi-squared test was used to evaluate associations among the variables, adopting a significant level of 0.05. RESULTS: Females predominated, with 1.4 women for each man. The patients mean age was 58.9 years. Nonwhite patients represented 13.7% of the sample. The prevalent anatomical sites for cutaneous melanoma were the trunk and feet, for both men and women. Acral lentiginous melanoma represented 22.3% of the cohort. In situ primary lesions were observed in few cases and a high percentage of thick cutaneous melanoma was detected. Ulceration was found in 13.4% of the thin tumors (< 1.0 mm). Thicker and ulcerated lesions predominated in male patients (p = 0.011 and p < 0.001 respectively) and in elderly patients (p = 0.021 and p = 0.015). CONCLUSIONS: The cohort mostly presented thick and ulcerated tumors, denoting late diagnosis and bad prognosis. Also, the sample was characterized by considerable prevalence of female patients, nonwhite patients, limb lesions and acral lentiginous melanoma. PMID- 18425287 TI - Sudden infant death syndrome in Brazil: fact or fancy? AB - CONTEXT AND OBJECTIVE: The true incidence of sudden infant death syndrome (SIDS) in Brazil is unknown. The aim here was to identify SIDS cases in the city of Ribeirao Preto, State of Sao Paulo, between 2000 and 2005, in order to estimate its incidence. DESIGN AND SETTING: Retrospective analysis of data on live births and infant deaths in Ribeirao Preto and from autopsies of infants performed at the Death Verification Service of the Interior (SVOI) between 2000 and 2005. RESULTS: There were 47,356 live births and 537 deaths, with infant mortality rates ranging from 12.9 to 10.9 of live births. Among the 24 infants who died possibly due to SIDS and who were autopsied at the SVOI, six were from families living in the municipality (0.13 of live births): three (50%) were diagnosed as SIDS, and one each (16.66%) as indeterminate cause, bronchoaspiration and cerebral edema. Two deaths occurred in the first month of life (33.33%) and one each (16.66%) at two, four, six and eight months. Two deaths each (33.33%) occurred in the months of February and December, one each in August and October (16.66%). Four cases (66.7%) occurred in the summer and one each (16.66%) in winter and spring. There was 5:1 predominance of males over females. CONCLUSIONS: The frequency of SIDS was lower than what has been reported worldwide and in the Brazilian literature, thus suggesting underdiagnosis, indicating the lack of any specific postmortem protocol for SIDS identification and showing the need to implement this. PMID- 18425288 TI - Diagnosis and treatment of polycythemia vera: Brazilian experience from a single institution. AB - CONTEXT AND OBJECTIVE: Polycythemia vera (PV) is a chronic myeloproliferative disorder characterized by predominant proliferation of erythroid precursors. Few data are available concerning Brazilian patients with this condition. The aim of this study was to describe clinical and demographic characteristics of PV patients at diagnosis and analyze their long-term outcomes. DESIGN AND SETTING: Retrospective study at the Division of Hematology, Faculdade de Medicina da Universidade de Sao Paulo (FMUSP), Sao Paulo. METHODS: All consecutive patients with PV diagnosed according to World Health Organization criteria were eligible for this study. Clinical and demographic characteristics, thrombotic events, transformation to acute leukemia, myelofibrosis and survival were evaluated. RESULTS: Sixty-six patients were evaluated. Thirty-six (54.5%) were females, with a median age at diagnosis of 61 years. At diagnosis, the median hemoglobin concentration was 18.8 mg/dl and the median platelet count was 593,000/mm(3). Fifty-eight patients (88.0%) were treated with hydroxyurea with or without phlebotomy. During a median follow-up of 77 months, 22 patients (33.3%) had new thrombotic events, mainly of arterial type. The overall incidence of leukemia and myelofibrosis was 0.42% per patient-year and 1.06% per patient-year, respectively. Median overall survival was not reached and the seven-year survival rate was 77.8%. CONCLUSION: The PV patients described here had long survival and arterial thrombotic events were the most important and common complication among this population. PMID- 18425289 TI - Demographics and complaints of university students who sought help at a campus mental health service between 1987 and 2004. AB - CONTEXT AND OBJECTIVE: Client characterization is an important step in evaluating the services offered by campus counseling and mental health centers and in their further planning and development. The objectives here were to describe reported complaints and demographics among students who sought counseling/mental healthcare at a Brazilian campus mental health service over a 17-year period and to compare these characteristics with those of the general university student body. DESIGN AND SETTING: Retrospective study at the Psychological and Psychiatric Service for Students (SAPPE), Universidade Estadual de Campinas (Unicamp). METHODS: The participants were all of the 2,194 students who sought counseling/mental health care at SAPPE from 1987 to 2004. Information was obtained from clients clinical charts. Unicamps database was consulted for general information on its students. RESULTS: The findings indicated overrepresentation, among the clients, of undergraduates, female students, students from Brazilian states other than Sao Paulo, students living in the campus residence hall and those whose main source of income was a scholarship grant. We also found overrepresentation of Humanities and Arts students among the clients. The most frequently reported complaints were difficulties in interpersonal relationships, family conflicts and poor academic performance. CONCLUSION: Course level (undergraduate or postgraduate), study field, living in a university residential facility and reliance on a scholarship grant were found to influence the behavior of seeking mental health counseling among Brazilian university students in this study. Course level was found to influence the pattern of complaints reported at first contact with the mental health service. PMID- 18425290 TI - Low-cost outpatient chemotherapy regimen of cisplatin, 5-fluorouracil and leucovorin for advanced head and neck and esophageal carcinomas. AB - CONTEXT AND OBJECTIVE: Patients with advanced head and neck (H/N) and esophageal squamous cell carcinoma (SCC) often have a poor performance status and a dire prognosis. Our aim was to evaluate the feasibility, activity and quality of life (QOL) of an outpatient chemotherapy regimen consisting of cisplatin, 5 fluorouracil and leucovorin (CFL). DESIGN AND SETTING: Prospective phase II study conducted at a Brazilian public institution. METHODS: Fifteen patients with residual, recurrent or metastatic SCC of the H/N or esophagus received bolus infusions of leucovorin 20 mg/m(2)/day and 5-fluorouracil 370 mg/m(2)/day on days 1-4, and 90 minutes of infusion of cisplatin 25 mg/m(2)/day on days 1-3, every 21 to 28 days, depending on hematological recovery. We also evaluated QOL by applying the European Organization for Research and Treatment of Cancer Quality of Life-C30 questionnaire (EORTC QLQ-C30) before each cycle. RESULTS: The overall response rate was 36%, and the mean overall survival and progression-free survival were six and three months, respectively. We observed grade 3 or higher hematological toxicity in seven patients and one patient had grade 3 nausea and vomiting. One patient died because of neutropenic fever. Seven out of the 12 patients who could be evaluated regarding QOL presented an improvement in their overall health status and functional QOL scores over the course of the treatment. CONCLUSIONS: CFL is an active outpatient protocol with tolerable toxicity and a favorable QOL impact. Larger studies are warranted, in order to confirm these results. PMID- 18425291 TI - Comments on the manuscript "Hepatorenal syndrome: an update". PMID- 18425292 TI - [Cultural meanings of infantile asthma]. AB - OBJECTIVE: To analyze the cultural meanings of infantile asthma from the perspective of the mother/carer of the child. METHODOLOGICAL PROCEDURES: Qualitative research conducted in 2004, in the city of Fortaleza (Northeastern Brazil). An ethnographic approach was utilized, consisting of participant observation and interviews with seven mothers, accompanying their children in a hospital emergency ward. Thematic analysis was the technique employed when identifying the meanings of mothers' discourse. ANALYSIS OF RESULTS: Two discussion categories were identified: "disinformation on illness" in which it was possible to perceive that the mothers were not informed with respect to their children's illness; and "cultural care" in which they relate information on caretaking and they use resources of popular knowledge to prevent their children's asthma, such as environmental care and the use of popular remedies, based on medicinal plants. CONCLUSIONS: The disinformation and lack of maternal knowledge on infantile asthma among the mothers indicate the need for an intense educational program, problematizing in nature and based on dialogue, conducted in close collaboration with the treatment aimed at improving the prognosis of the disease. PMID- 18425293 TI - [Food frequency questionnaire for adults from a population-based study]. AB - The objective of the study was to develop a food frequency questionnaire for each of the following groups: female adults, male adults and male and female adults, based on dietary data obtained in a population-based study comprising individuals from different income levels. Dietary data was obtained from a 24-hour Dietary Recall applied to a probabilistic sample of 1,477 subjects in the city of Sao Paulo (Southeastern Brazil) in 2003. Food items accounting for at least 90% of total daily calorie intake and nutrients were selected. The reference time frame was the year preceding the interview and subjects had the choice of four serving sizes. PMID- 18425294 TI - Seroprevalence of human parvovirus B19 in a suburban population in Sao Paulo, Brazil. AB - OBJECTIVE: To analyze the prevalence of IgG antibodies to human parvovirus B19. METHODS: Cross-sectional study in a suburban community in Sao Paulo, Southeastern Brazil, between November 1990 and January 1991. Randomly selected (N=435) representative samples of sera were collected from healthy children older than 15 days old and adults up to 40 years old. IgG antibodies were detected using ELISA. RESULTS: High prevalence of IgG antibodies to B19 parvovirus was found in 87% of newborns. The prevalence of maternally derived IgG antibodies exponentially plunged up to the 19th month of age. Low prevalence of antibodies was found in the first 4 years of life, increasing up to 72% in those aged 31-40 years. It was estimated that the average age of first infection in this population is 21 +/- 7 years old and the optimal age for vaccination with a hypothetical vaccine would be 1 year of age. CONCLUSIONS: Parvovirus B19 IgG antibody prevalence was high in newborns and those aged 31-40 years. The analysis by age groups showed a pattern similar to that found in previous studies, i.e., low prevalence of infection in children that increases with age. PMID- 18425296 TI - [Socioeconomic status and health: a discussion of two paradigms]. AB - Socioeconomic status and its impact on health are in the mainstream of public health thinking. This text discusses two paradigms utilized in assessing socioeconomic status in epidemiologic studies. One paradigm refers to prestige based measurements and positive differentiation among social strata. This paradigm is characterized by classifications assessing social capital and the access to goods and services. The other paradigm refers to the classification of social deprivation and negative differentiation among social strata. The proposal of State-funded reposition to the mostly deprived social strata is acknowledged as characteristic of this paradigm. The contrast between these paradigms, and their potential interaction and debate are discussed. Fostering reflection on methodological strategies to assess socioeconomic status in epidemiologic studies can contribute to the promotion of health and social justice. PMID- 18425295 TI - [Sex, income and level of education associated with physical activity level among workers]. AB - OBJECTIVE: To analyze socioeconomic and demographic factors associated with physical activity level among workers. METHODS: Cross-sectional study conducted in Brazil's Federal District, from 2000 to 2001, with a representative sample of 1,044 workers from both sexes, who are covered by the Programa de Alimentacao do Trabalhador (Workers' Food Program). Binary logistic regression was used to determine possible risk factors associated with physical activity level and selected by the backward elimination process. Interactive effects of the variables were verified in the final model by means of the odds ratio for physical activity level >1.4. RESULTS: The final model included sex, income and level of education. Male individuals who only finished elementary school and earned less than four Brazilian minimum wages per month were the most likely to show physical activity level >1.4. The odds ratio estimate indicated that men were five times more likely to show physical activity level >1.4 than women. Individuals who only finished elementary school and had an income equal to or below four minimum wages were two times more likely to show physical activity level >1.4 than individuals who earned more than four minimum wages and had a university degree or higher. CONCLUSIONS: Results indicate that the factors associated with physical activity level among workers are the following: male, elementary school level, and income below four Brazilian minimum wages per month. PMID- 18425297 TI - [Air quality and acute respiratory disorders in children]. AB - OBJECTIVE: To assess the relationship between air pollution and acute respiratory disorders in children. METHODS: A time series ecological study was carried out in three public health posts in a region of the city of Rio de Janeiro (Southeastern Brazil), between April 2002 and March 2003. Data for PM10, SO2, NO2, CO and O3 were analyzed daily and as closure variables, a total of 45,595 emergency pediatric consultations for respiratory symptoms or specifically for disorders in the upper and lower airways. To control for confounders, other variables were included in the model including those relating to weather, seasonality, temperature, relative air humidity, rain volume, respiratory infections and the calendar effects (such as holidays and weekends). A Poisson regression was applied using generalized counting models to estimate the effects of pollutants and confusing factors. RESULTS: Only O3 had a positive and statistically significant effect, both among emergency consultations for respiratory problems and consultations for symptoms relating to the lower airways. Effect and exposure occurred on the same day (lag 0). A significant negative association was found with CO and pediatric consultations for respiratory complaints. Other air pollutants were not found to have a significant effect. CONCLUSIONS: There were found associations between outdoor air pollution and the number of emergency pediatric consultations for respiratory problems in the studied area, in spite of the fact that the levels of all pollutants monitored during the study period were below recommended levels. PMID- 18425298 TI - Supporting the diagnosis of non-specific respiratory symptoms in primary care: the role of exhaled nitric oxide measurement and spirometry. AB - AIMS: To assess whether exhaled nitric oxide (FENO) measurements improve management and clinician confidence in patients presenting with non-specific respiratory symptoms. METHODS: This observational study was based in a large primary care practice (15,500 patients, 14 GPs). Patients had non-specific respiratory symptoms for at least six weeks. FENO and spirometry measurements were performed at initial assessment. An algorithm was employed to assist interpretation of FENO and spirometry results. GPs evaluated the diagnostic contribution of FENO and spirometry at 3- month follow-up. RESULTS: In 48/51 (94%) of cases FENO was considered significant in formulating a diagnosis. Spirometry was deemed helpful in 27/51 (54%). CONCLUSION: FENO measurements improved diagnostic confidence when assessing non-specific respiratory symptoms. This may be because, in contrast to spirometry, both low and high FENO values have clinical significance. TRIAL REGISTRATION: Australian Clinical Trials Registry ACTRN012605000354684. PMID- 18425299 TI - The use of omalizumab in asthma. AB - Asthma causes a substantial burden of morbidity and mortality, affecting 300 million people worldwide - a figure predicted to increase to 400 million by 2025. Despite the availability of a variety of treatment options and detailed treatment guidelines, many patients with asthma, and in particular those with severe persistent asthma, remain inadequately controlled. Approximately 50-80% of severe asthma has an allergic component, with immunoglobulin E (IgE) playing a role in the underlying allergic inflammatory cascade. Omalizumab is a humanised monoclonal anti-IgE antibody that targets IgE and partially inhibits the inflammatory cascade. Clinical trials have demonstrated that omalizumab added to standard asthma therapy reduces exacerbations and emergency visits with concomitant improvements in asthma control and quality of life in patients with moderate-to-severe and severe persistent allergic (IgE-mediated) asthma. Add-on omalizumab is indicated for the treatment of patients with inadequately controlled moderate-to-severe (US label) and severe (EU label) persistent allergic asthma despite treatment with high-dose inhaled corticosteroids (and in the EU, high-dose inhaled corticosteroids plus a long-acting beta2-agonist). Within this highly-targeted patient population, analyses have been unable to identify pre-treatment clinical characteristics that are predictive of a greater response to omalizumab. In contrast, assessment of response to omalizumab following 16 weeks of treatment appears to be reliably judged by physicians in clinical trial settings and may be a feasible means of selecting patients who should continue treatment. PMID- 18425300 TI - Systems for the management of respiratory disease in primary care - an international series: Canada. AB - Canada has a universal health care system funded by the government. All people are supposed to have unrestricted access to all essential health care in a timely fashion. Canada has ten provinces and three territories. Health care is funded by each province/territory, with federal payments providing some of the funding. The bulk of the provision of respiratory care in Canada is provided by primary care practitioners. Across the country we have a family physician shortage; thus, in many areas of the country there is insufficient access since patients do not actually have a family physician. This has less effect on acute medical services, which can be available in ER or walk-in settings, but certainly does affect the provision of services for chronic illnesses. While thus far the health care system is 'free', there are some significant limitations; the commonest is waiting times for specialist care and investigations. Other significant deficiencies include the lack of coverage for medications for both acute and chronic conditions and of medical devices. Primary care reforms by local governments have attempted to fix these issues by changing care models. Fee-for service medicine by physicians is slowly being changed in places to capitation models and other systems such as rewards for managing chronic conditions optimally. Ontario has instituted 'reward systems' for diabetes and smoking cessation. British Columbia has rewards for multiple chronic diseases. In many areas, care in the provinces is regionalised to allow local arrangements to help decide on where and how care is organised. Respiratory diseases (excluding lung cancer) rank fourth in Canada in the total proportion of direct health care costs (behind neuropsychiatric, injury and cardiovascular diseases). A number of studies have shown that respiratory conditions such as asthma and COPD are underdiagnosed and/or undermanaged. Other conditions need treatment by specialists or physicians with a special interest (TB, cystic fibrosis, lung cancer) and access to these physicians also is a barrier to health care provision. Health care systems are thus part, but not all, of the problem. PMID- 18425301 TI - Protective effect of polydatin against ischemia/reperfusion injury in rat heart. AB - The aim of the present study was to investigate the protective effect of polydatin against myocardial ischemia/reperfusion injury in rats and the underlying mechanism. In anesthetized rats, ischemia and reperfusion arrhythmia produced by ligating and loosing the coronary artery was recorded and myocardial infarct size was measured. In Langendorff isolated rat heart, cardiac function was recorded before and after 30 min of global ischemia followed by 60 min of reperfusion. The parameters of cardiac function include left ventricular developed pressure (LVDP), maximal differentials of LVDP (+/-LVdp/dt(max)) and coronary flow (CF) were measured. Myocardial superoxide dismutase (SOD) activity, the contents of myocardial malondialdehyde (MDA) and nitric oxide (NO) as well as the activity of nitric oxide synthase (NOS) were measured in isolated heart. The results showed: (1) Arrhythmia score and myocardial infarct size were significantly lower in polydatin group than that in the control group (P<0.05, P<0.01); (2) The recovery of LVDP, +/-LVdp/dt(max) and CF during reperfusion in polydatin group were significantly better than that in the control rats (P<0.05, P<0.01); (3) SOD activity in polydatin group was significantly higher than that in the control group, but MDA content was lower in polydatin group than that in the control group (P<0.05); (4) NO content and NOS activity, especially constitutive nitric oxide synthase (cNOS) activity in polydatin group were higher than that in the control group (P<0.05); (5) L-NAME, the NOS inhibitor, reversed the protective effect of polydatin against ischemia/reperfusion injury. The results suggest that polydatin has a protective effect against ischemia/reperfusion injury in rat heart. The cardioprotection of polydatin is mainly mediated by cNOS which leading to an increase in NO production. PMID- 18425302 TI - [Nongenomic action and mechanism of 17beta-estradiol in cytosolic calcium concentration in delayed implantation mouse endometrial stromal cells]. AB - To investigate the existence of nongenomic action of 17beta-estradiol (E(2)) in the delayed implantation mouse endometrial stromal cells, the changes in intracellular calcium concentration ([Ca(2+)](i)) and the upstream of calcium signal in vitro were detected. The experiment was composed of two parts. Firstly, the change in [Ca(2+)](i) in endometrial stromal cells induced by E(2) under different conditions was detected. The mice were divided into 6 groups as follows: 0.1% dimethylsulfoxide (DMSO) control group, 1*10(-8) mol/L bovine serum albumin (BSA) control group, 1*10(-8) mol/L E(2) group, 1*10(-8) mol/L E(2) conjugated with BSA (E(2)-BSA) group, 1*10(-8) mol/L E(2) + calcium-free medium group, 1*10(-8) mol/L E(2) + 5 mg/mL tamoxifen group, with 4 mice in each group. The endometrial tissue was obtained from delayed implantation mice at pregnant day 7, and digested by incubation of tissue minces in Hankos balanced salts (HBSS, pH 7.2), which contained glucose (1 g/L), and collagenase I (0.125%), for 1 h at 37 degrees C. The stromal cells were preloaded with 2.5 mmol/L Fluo-3/AM, a fluorescent probe of calcium, for 30 min. A confocal laser scanning microscope, which fixed the wave length of excitation and emission at 488 nm and 526 nm, respectively, was used to detect the change in [Ca(2+)](i). Secondly, the mechanism of E(2) effects in endometrial stromal cells was investigated. Immunofluorescent method was used to detect the change in phosphorylation of phospholipase C (PLC) before and after the stromal cells were treated with E(2) for 5 min, 15 min, and 30 min. Seven delayed implantation mice were used. The results were as follows. [Ca(2+)](i) increased immediately and reached the maximum at 15 min after the stromal cells were treated with 1*10(-8) mol/L E(2) and returned to the normal level at 30 min. In E(2)-BSA group and E(2) + calcium free medium group the same results were obtained as that in E(2) group, but there was no increase of [Ca(2+)](i) in DMSO and BSA groups. Tamoxifen, a traditional antagonist of estrogen receptor, did not inhibit the increase in [Ca(2+)](i) induced by E(2). Immunofluorescent results showed that the change in phosphorylated-PLC level had the same trend as [Ca(2+)](i) after the cells were treated with E(2). Compared with that in the control group, the immunofluorescent intensity increased at the beginning and achieved the maximum at 15 min (P<0.001), then declined to the normal level at 30 min. These results suggest that the rapid response of [Ca(2+)](i) induced by E(2) in the endometrial stromal cells in delayed implantation mice is possibly carried out through a nongenomic pathway, and the transmembrane signal transduction is related to the phosphorylation of PLC in this process. PMID- 18425303 TI - Electrophysiological effects of hydrogen sulfide on pacemaker cells in sinoatrial nodes of rabbits. AB - The cardiac electrophysiological effects of hydrogen sulfide (H(2)S) on pacemaker cells in sinoatrial (SA) nodes of rabbits were examined using intracellular microelectrode technique. The results obtained were as follows: (1) The velocity of diastolic (phase 4) depolarization (VDD) and rate of pacemaker firing (RPF) in normal pacemaker cells in SA nodes were decreased by NaHS (H(2)S donor) (50, 100, 200 MUmol/L) in a concentration-dependent manner; (2) ATP-sensitive K(+) (K(ATP)) channel blocker glybenclamide (Gli, 20 MUmol/L) blocked the effect of NaHS (100 MUmol/L) on pacemaker cells; (3) Pretreatment with CsCl (2 mmol/L), a blocker of pacemaker current (I(f)), did not affect the effect of NaHS (100 MUmol/L) on SA node pacemaker cells; (4) DL-propargylglycine (PPG, 200 MUmol/L), an inhibitor of cystathionine gamma-lyase (CSE), did not affect the parameters of action potentials in pacemaker cells in SA nodes. All these results suggest that H(2)S exerts a negative chronotropic action on pacemaker cells in SA nodes of rabbits. These effects are likely due to an increase in potassium efflux through opening K(ATP) channels; I(f)is unlikely to play a major role in these effects. In our study, there was no evidence for the generation of endogenous H(2)S by CSE in SA node pacemaker cells. PMID- 18425304 TI - Role of transient receptor potential vanilloid 4 in the effect of osmotic pressure on myocardial contractility in rat. AB - The aim of the present study was to investigate the influence of osmotic pressure on myocardial contractility and the possible mechanism. Electrical stimulation was used to excite papillary muscles of the left ventricle of Sprague-Dawley (SD) rats. The contractilities of myocardium in hyposmotic, isosmotic, and hyperosmotic perfusates were recorded. The influences of agonist and antagonist of the transient receptor potential vanilloid 4 (TRPV4) on the contractility of myocardium under hyposmotic, isosmotic and hyperosmotic conditions were observed. The results were as follows: (1) Compared with that under isosmotic condition (310 mOsm/L), the myocardial contractility was increased by 11.5%, 21.5% and 25.0% (P<0.05) under hyposmotic conditions when the osmotic pressure was at 290, 270 and 230 mOsm/L, respectively; and was decreased by 16.0%, 23.7% and 55.2% (P<0.05) under hyperosmotic conditions when the osmotic pressure was at 350, 370 and 390 mOsm/L, respectively. (2) When ruthenium red (RR), an antagonist of TRPV4, was added to the hyposmotic perfusate (270 mOsm/L), the positive inotropic effect of hyposmia was restrained by 36% (P<0.01); and when RR was added to the hyperosmotic perfusate (390 mOsm/L), the inhibitory effect of hyperosmia on myocardial contractility was increased by 56.1% (P<0.01). (3) When 4-alpha phorbol-12,13-didecanoate (4alpha-PDD), an agonist of TRPV4, was added to the isosmotic perfusate (310 mOsm/L), the myocardial contractility did not change; and when 4alpha-PDD was added to the hyperosmotic perfusate (390 mOsm/L), the inhibition of myocardial contractility by hyperosmia was increased by 27.1% (P<0.01). These results obtained indicate that TRPV4 is possibly involved in the osmotic pressure-induced inotropic effect. PMID- 18425305 TI - [Effect of phalloidin on electrophysiological changes induced by stretch of myocardial infarcted hearts in rats]. AB - The present study aimed to explore whether the stretch of ischemic myocardium could modulate the electrophysiological characteristics via mechanoelectric feedback (MEF), as well as the effect of phalloidin on the electrophysiological changes. Thirty-two Wistar rats were randomly divided into 4 groups: control group (n=9), phalloidin group (n=7), myocardial infarction (MI) group (n=9), MI + phalloidin group (n=7). The acute myocardial infarction (AMI) was conducted by ligation of the left anterior descending (LAD) coronary artery for 30 min in isolated rat heart. The volume alternation of a water-filled latex balloon in the left ventricle produced the stretch of myocardium. After perfused on Langendorff, the isolated hearts were stretched for 5 s by an inflation of 0.1, 0.2 and 0.3 mL separately and the effect of stretch was observed for 30 s, including the left ventricular systolic pressure (LVSP), left ventricular end-diastolic pressure (LVEDP), +/-dp/dt(max), monophasic action potential duration at 90% repolarization (MAPD90), and occurrence of premature ventricular beats (PVB) and ventricular tachycardia (VT). The stretch caused an increase of MAPD(90) in both control and MI rats (P<0.05, P<0.01). Moreover, MAPD(90) in MI group increased more significantly than that in the control group at the same degree of stretch (P<0.05, P<0.01). Phalloidin (1 MUmol/L) had no effect on MAPD(90) in basal state. After stretch, MAPD(90) in phalloidin group slightly increased but was not significantly different from that in the control group. However, phalloidin reduced MAPD(90) in infarcted myocardium, especially when DeltaV=0.3 mL (P<0.05). The incidence rates of PVB and VT in MI group were higher than that in the control group (both P<0.01). And there was no significant difference in the incidence rates of PVB and VT between phalloidin group and control group. Phalloidin inhibited the occurrence of PVB and VT in infarcted hearts (both P<0.01). LVSP and +dp/dt(max) in MI group obviously decreased (P<0.01 vs control). With application of phalloidin, LVSP slightly, but not significantly increased in infarcted hearts, while -dp/dt(max) significantly increased (P<0.05). It is suggested that MI facilitates the generation and maintenance of malignant arrhythmias, while phalloidin obviously inhibits the occurrence of arrhythmias. PMID- 18425306 TI - [Protein kinase C regulates unloading contraction of cardiomyocytes in hypertrophic heart of spontaneously hypertensive rat]. AB - The activation of protein kinase C (PKC) is not only a pivotal node during cardiac hypertrophy in chronic pressure-overloaded heart, but also involved in the regulation of cardiac contractility. The aim of this paper was to observe PKC modulation in cardiac contractility at different stages of cardiac hypertrophy in spontaneously hypertensive rat (SHR). The cardiomyocytes were isolated from 4- and 10-month-old normotensive Wistar-Kyoto (WKY) and SHR rat hearts. The shortening amplitude of unloading contraction in cardiomyocytes was observed by an Edge Detector system. The shortening amplitude in WKY rat cardiomyocytes increased gradually as the stimulating frequency increased from 1 to 3 Hz. The shortening amplitude was positively correlated with stimulating frequency. The shortening amplitude in 4-month-old SHR group was enhanced as compared with that in WKY group at the same stimulating frequency. When the stimulating frequency increased, the shortening amplitude did not increase in 4-month-old SHR group. There was no difference in shortening amplitude between 10-month-old SHR and WKY groups at 1-Hz stimulating frequency. But the shortening amplitude in 10-month old SHR group decreased when the stimulating frequency increased to 3 Hz. The perfusion of 50, 100 or 200 nmol/L PMA (a non-specific agonist of PKCs) significantly reduced the shortening amplitude in WKY and SHR groups. The shortening amplitude was reduced to (69.8+/-1.9)%, (58.2+/-2.2)% and (22.7+/ 2.5)% (all P<0.01), respectively, in WKY group, as compared with that in HEPES buffer perfusion (100%). It was reduced to (6.1+/-0.7)%, (2.4+/-0.2)% and (12.5+/ 2.6)% (all P<0.01) in 4-month-old SHR group, and (65.7+/-1.6)%, (53.9+/-4.0)% and (16.3+/-2.0)% (all P<0.01) in 10-month-old SHR group. The decreases in shortening amplitude in 4-month-old SHR group were more significant than those in 10-month old SHR and WKY groups. On the other hand, 200 nmol/L of staurosporine, an inhibitor of PKC, significantly increased the shortening amplitude of cardiomyocytes in WKY, 4-month-old SHR, and 10-month-old SHR groups by (63.63+/ 4.53)%, (80.82+/-4.61)% and (80.97+/-4.59)%, respectively (all P<0.05). The results suggest that the PKC isoforms inducing negative inotropic effect may be activated at the early stage of cardiac hypertrophy in SHR rats, and are possibly involved in the modulation of cardiac contractility. The activated PKC isoforms return to their normal activity at the stable stage of cardiac hypertrophy in SHR rats. PMID- 18425307 TI - [Activity identification of peptide containing rat sodium pump alpha2 subunit M1 M2 extramembrane fragment in vitro]. AB - In order to explore the activity of a peptide containing rat sodium pump alpha2 subunit M1-M2 extramembrane fragment (RES2 derivative) in vitro, the peptide (Leu Ala-Ala-Met-Glu-Asp-Glu-Pro-Ser-Asn-Asp-Asn-Gly-Gly-Gly-Ser) was synthesized by peptide synthesizer with Fmoc method and purified by high performance liquid chromatography (HPLC). Its binding activity was identified by radioligand receptor binding assay (RRA) and its bioactivity was measured by erythrocyte (86)Rb uptake. The results of saturation binding experiment and competitive binding experiment showed that the synthesized RES2 derivative had the capability to bind to (3)H-ouabain. The dissociation constant (K(d)) was 38.46 nmol/L and IC(50) was 6.353 nmol/L. Erythrocyte (86)Rb uptake experiment showed that the RES2 derivative blocked the inhibitory effect of ouabain on the sodium pump on erythrocyte membrane in a dose-dependent manner. The results showed that the RES2 derivative is capable of binding to ouabain and improving the activity of sodium pump on erythrocyte membrane, suggesting that the RES2 derivative might become an effective antihypertensive drug in the future. PMID- 18425308 TI - [Role of endogenous hydrogen sulfide in pulmonary hypertension induced by lipopolysaccharide]. AB - The purpose of the present study was to explore the role of endogenous hydrogen sulfide (H2S) in pulmonary arterial hypertension induced by endotoxin. Adult male Sprague-Dawley (SD) rats were randomly divided into four groups: Control group (0.5 mL/kg body weight of normal saline, i.v.), lipopolysaccharide (LPS)-treated group (5 mg/kg body weight of LPS, i.v.), LPS + NaHS (5 mg/kg body weight of LPS, i.v., and 28 MUmol/kg body weight of NaHS, i.p.) and LPS + PPG group (5 mg/kg body weight of LPS, i.v., and 30 MUmol/kg body weight of PPG, i.p.). Rats were anesthetized with 20% urethane (1 g/kg body weight, i.p.). A polyethylene catheter was inserted into the pulmonary artery through the right external jugular vein to measure the mean pulmonary arterial pressure (mPAP) for 7 h, and then the pulmonary artery was isolated rapidly by the method described previously. Pulmonary arterial activity was detected. H2S concentration and cystathionine gamma-lyase (CSE) activity in pulmonary artery tissues were determined by biochemical method. CSE mRNA expression was detected by competitive reverse transcriptase-polymerase chain reaction (RT-PCR). Compared with control, LPS significantly increased mPAP [(1.82+/-0.29) kPa vs (1.43+/-0.26) kPa, P<0.01], decreased H2S production [(26.33+/-7.84) vs (42.92+/-8.73) pmol/g wet tissue per minute, P<0.01), and reduced endothelium-dependent relaxation response [(75.72+/-7.22)% vs (86.40+/-4.40) %, P<0.01) induced by ACh (1*10(-6) mol/L). These effects were partly reversed by co-administration of NaHS and enhanced by co-administration of PPG. Both CSE activity and CSE mRNA expression were consistent with H2S production. It is suggested that the inhibitory effect of LPS on endothelium-dependent relaxation results in pulmonary hypertension, which might be mediated through H(2)S. PMID- 18425309 TI - [Role of 5-HT(2A) receptor in increase in respiratory-related rhythmic discharge activity by nikethamide in neonatal rat transverse medullary slices]. AB - To investigate the effects of nikethamide on the generation and modulation of rhythmic respiration of neonatal rats and the role of 5-HT(2A) receptor in this course, experiments were performed on the transverse medullary slices of neonatal rats (both sexes, 1-3 d) in vitro. The slices containing the medial region of the nucleus retrofacialis (mNRF) with the hypoglossal nerve rootlets were prepared in which the respiratory-related rhythmic discharge activity (RRDA) was recorded from the hypoglossal nerve rootlets by suction electrode. The possible role of nikethamide on RRDA was investigated by administration of an agonist of 5-HT(2A) receptor, 1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane (DOI), and an antagonist of 5-HT(2A) receptor, ketanserine, dissolved in modified Krebos solution (MKS). Thirty slices were randomly divided into five groups: Group 1: the slices were perfused with different concentrations of nikethamide (0.5, 1, 3, 5, 7, 10 MUg/mL), and the most effective concentration was selected; Group 2: the slices were perfused with DOI (40 MUmol/L); Group 3: the slices were perfused with ketanserine (40 MUmol/L); Group 4: the slices were perfused with ketanserine + DOI; Group 5: the slices were perfused with nikethamide, then perfused with nikethamide + ketanserine after washout of nikethamide. Nikethamide increased RRDA in transverse medullary slices at 0.5-7 MUg/mL, and 5 MUg/mL was the most effective concentration. DOI increased RRDA with prolonged inspiratory time (TI), increased integral amplitude (IA), and shortened respiratory cycle (RC). Ketanserine decreased RRDA with shortened TI, decreased IA and prolonged RC. Ketanserine + DOI had no significant effects on RRDA. The effects of nikethamide on RC and IA were totally and partially reversed by additional application of ketanserine, but the effect of nikethamide on TI was not influenced by ketanserine. It is proposed that nikethamide increases RRDA partly via 5-HT(2A) receptors. PMID- 18425310 TI - H(2)S protects myocardium against ischemia/reperfusion injury and its effect on c Fos protein expression in rats. AB - The present study was aimed to study the effect of hydrogen sulfide (H(2)S) on rat myocardial ischemia/reperfusion (I/R) injury and whether the effect is mediated by c-Fos protein expression. Male Sprague-Dawley rats were randomly divided into 4 groups: CONTROL GROUP: sham treatment; I/R group: the rat anterior descending branch of left coronary artery was occluded for 30 min and then released to allow reperfusion for 60 min; NaHS (exogenous H(2)S donor) groups: the rats were pretreated with NaHS at 2.8 MUmol/kg body weight and 14 MUmol/kg body weight (i.v.), respectively, before I/R treatment. Hemodynamics (LVSP, LV+/ dp/dt(max)) and electrocardiogram (ECG, lead II) were monitored continuously with multi-channel physiological signal analysis system after reperfusion. Myocardial infarct size was measured using triphenyltetrazolium chloride (TTC) staining. H(2)S concentration in the plasma was determined with a spectrophotometer. Morphological and ultrastructural changes in myocardial tissue were evaluated by HE staining and by a transmission electron microscope. The evaluation of c-Fos protein expression in myocardial tissue was performed by immunohistological staining. The results showed that H(2)S concentration in rat plasma in I/R group was significantly decreased compared with that in the control group [(30.32+/ 5.26) vs (58.28+/-7.86) MUmol/L, P<0.05]. NaHS at 2.8 and 14 MUmol/kg body weight reduced the changes in LVSP, LV+/-dp/dt(max) in rat myocardium induced by I/R injury. The values of LVSP, +dp/dt(max) and -dp/dt(max) at 60 min during myocardial reperfusion were enhanced from (75.93+/-1.10)%, (66.27+/-4.78)% and (66.01+/-4.79)% in I/R group to (84.34+/-2.24)%, (76.38+/-1.93)% and (75.47+/ 5.29)% in 2.8 MUmol/kg body weight NaHS group (P<0.05, P<0.01, n=6), (88.40+/ 2.88)%, (80.10+/-2.09)% and (80.48+/-6.20)% in 14 MUmol/kg body weight NaHS group (P<0.01, n=6), respectively. Compared with that in 2.8 MUmol/kg body weight NaHS group, the enhancing effect was more prominent in 14 MUmol/kg body weight NaHS group. NaHS at 14 MUmol/kg body weight markedly alleviated the injury in morphological changes and decreased c-Fos protein expression in myocardial tissue compared with that in I/R group (0.20+/-0.06 vs 0.32+/-0.10, P<0.05). These results suggest that H(2)S protects myocardium against I/R injury and this protective effect may be related to the down-regulation of c-Fos protein expression. PMID- 18425311 TI - [Effect of Rhizoma curcumae oil on the learning and memory in rats exposed to chronic hypoxia and the possible mechanisms.]. AB - The effect of Rhizoma curcumae oil on the learning and memory in rats exposed to chronic hypoxia and the possible mechanisms were investigated. The rats were divided randomly into 5 groups (14 animals in each group): control, chronic hypoxia, chronic hypoxia with low (5 mg/kg body weight), middle (10 mg/kg body weight) and high (20 mg/kg body weight) concentrations of Rhizoma curcumae oil injection. The animals undergoing chronic hypoxia were exposed to hypoxia in a hypoxic chamber containing 10% O(2) and 5% CO(2) for 10 h/d, lasting 28 d. Morris water maze (MWM) test was used to obtain the scores of leaning and memory. The superoxide dismutase (SOD) activity and malonaldehyde (MDA) content were determined in the serum and hippocampus as well as [Ca(2+)](i) in the hippocampus. The expression of phosphorylated Ca(2+)/calmodulin-dependent protein kinase II (p-CaMKII) in the hippocampus was evaluated by using immunohistochemistry and Western blot. Compared with the control group, the chronic hypoxia group showed the following changes: (1) The escape latency to the hidden platform was remarkably prolonged (P<0.05); (2) The content of MDA and [Ca(2+)](i) were obviously higher, but the activity of SOD and the expression of p-CaMKII were significantly lower (P<0.05, P<0.01). Compared with the chronic hypoxia group, groups with Rhizoma curcumae oil injection had the following changes: (1) The escape latency to the hidden platform was remarkably shorter in 10, 20 mg/kg body weight groups (P<0.05); (2) The content of MDA and [Ca(2+)](i) were markedly decreased in 5, 10, 20 mg/kg body weight groups (P<0.05, P<0.01), but the activity of SOD in the serum and the expression of p-CaMKII were significantly higher in 10, 20 mg/kg body weight groups (P<0.05, P<0.01). The results showed that the capacity of learning and memory was degraded following chronic hypoxia. The decrease in MDA content and [Ca(2+)](i) and (or) the increase in SOD activity and p-CaMKII expression might participate in the enhancing effect on learning and memory induced by Rhizoma curcumae oil. PMID- 18425312 TI - [Protection of hepatocyte growth factor on neurons subjected to oxygen-glucose deprivation/reperfusion]. AB - The present study was conducted to investigate the effect of hepatocyte growth factor (HGF) on cortical neurons exposed to oxygen-glucose deprivation/reperfusion (OGD/R). Primary cultured cerebral cortical neurons were prepared from Sprague-Dawley rats. The cells were used for experiments after culture for 12 d in vitro. To initiate OGD/R, the culture medium was replaced by glucose-free medium, and cells were transferred to a humidified incubation chamber flushed by a gas mixture of 95% N(2) and 5% CO(2) at 37 degrees C for 2 h. Following this treatment, neurons were fed with glucose-supplemented (25 mmol/L) medium, and returned to the incubator under normoxic condition for 0-24 h. The cell viability was assessed by MTT assay, and cell injury was evaluated by lactate dehydrogenase (LDH) leakage rate. The percentage of apoptotic cells was analyzed by flow cytometry and Hoechst 33258 staining. The expressions of c-Met mRNA and protein were detected by RT-PCR and Western blot analysis, respectively. Oxygen-glucose deprivation for 2 h decreased the cell viability and increased LDH leakage rate in cultured cerebral cortical neurons. The cell viability declined and LDH leakage rate increased with the reperfusion time going on (0-24 h). To explore the influence of HGF on neurons under oxygen-glucose deprivation for 2 h/reperfusion for 24 h (OGD(2)/R(24)) condition, the cultures were pretreated with HGF at different concentrations (5-120 ng/mL) 2 h prior to OGD(2)/R(24). The results showed that OGD(2)/R(24) treatment significantly decreased the cell viability, increased LDH leakage rate and the percentage of apopototic cells. Pretreatment with HGF at 5 ng/mL and 10 ng/mL did not affect the decrease in cell viability resulting from OGD(2)/R(24). In the presence of 20 ng/mL HGF, the increase in cell viability in cortical neurons exposed to OGD(2)/R(24) began to appear, and 80 ng/mL of HGF exhibited the maximal effect. HGF at 5, 10 and 20 ng/mL did not affect the increase in LDH leakage rate in cortical neurons exposed to OGD(2)/R(24). In the presence of 40 ng/mL HGF, the decrease in LDH leakage rate in cortical neurons subjected to OGD(2)/R(24) began to appear, and 80 ng/mL of HGF displayed the maximal effect. In addition, HGF at 80 ng/mL significantly attenuated cell apoptosis resulting from OGD(2)/R(24). As detected by semi quantitative RT-PCR and Western blot analysis, c-Met mRNA and protein were expressed in cerebral cortical neurons cultured for 12 d in vitro. c-Met mRNA and protein expressions in cortical neurons exposed to OGD(2)/R(24) were significantly upregulated and were not affected by pretreatment of HGF at 80 ng/mL. Treatment with c-Met inhibitor SU11274 (5 MUmol/L) completely eliminated HGF-mediated protection of cortical neurons subjected to OGD(2)/R(24). The results suggest that HGF directly protects cortical neurons against OGD/R-induced cell injury in a dose-dependent manner, and HGF has a potent anti-apoptotic action on neurons exposed to OGD/R. PMID- 18425313 TI - [Distribution and effects of 5-HT(1A) receptors in distal cerebral spinal fluid contacting neurons in rat brain parenchyma in neuropathic pain]. AB - The present study aimed to explore the effects of 5-HT(1A) receptors in the distal cerebral spinal fluid-contacting neurons (CSF-CNs) in rat brain parenchyma in neuropathic pain. The model of neuropathic pain with chronic constriction injury (CCI) of the sciatic nerve was made in Sprague-Dawley rats. The behavioral studies of animal were scored and the paw withdrawal latency (PWL) and paw withdrawal threshold (PWT) were measured. The distribution and expression of 5 HT(1A) receptors were observed in the distal CSF-CNs in brain parenchyma with double labeling of cholera toxin subunit B with horseradish peroxidase (CB-HRP) and 5-HT(1A) receptors with immunhistochemistry. The relationship between 5 HT(1A) receptors in distal CSF-CNs and neuropathic pain was analyzed. The results were as follows. On days 1, 3, 7, 14 of neuropathic pain, the PWL was 19.37+/ 2.74, 12.04+/-1.77, 8.74+/-1.15 and 12.31+/-1.94, respectively; the PWT was 18.58+/-3.62, 13.05+/-1.81, 6.66+/-1.43 and 11.55+/-2.01, respectively. CB-HRP labeled neurons of two clusters were always found in definite region but not in other area in brain parenchyma. The number of neurons double-labeled with CB HRP/5-HT(1A) receptors in each group was 276.14+/-36.00, 161.72+/-28.41, 108.64+/ 6.81, and 139.76+/-44.64, which was about 95%, 60%, 40% and 55% of all CB-HRP labeled neurons in the four courses of neuropathic pain, respectively. It is suggested that the distal CSF-CNs are always located in a special region in rat brain parenchyma and most of them have 5-HT(1A) receptors. A negative correlation is found between the expression of 5-HT(1A) receptors and neuropathic pain. PMID- 18425314 TI - [Changes in Bcl-2 and Caspase-3 expressions in cortex of hypoxic preconditioning mice]. AB - The purpose of the present study was to explore the roles of Bcl-2 and Caspase-3 in mouse cortex in hypoxic preconditioning. Blb/c mice were randomly divided into three groups: control group, hypoxic group and hypoxic preconditioning group. Fluorescence intensity of Bcl-2 and Caspase-3 was observed and number of positive cells was counted in parietal cortex by immunofluorescence and confocal laser scanning microscope. Fluorescence intensity of Bcl-2 in the normal group, hypoxic group and hypoxic preconditioning group was 6.2+/-1.7, 68.5+/-13.1, 180.6+/-34.8, respectively, and number of Bcl-2-positive cells was 18.5+/-4.9, 52.3+/-10.5, 150.8+/-24.7, respectively. Fluorescence intensity of Caspase-3 in the control group, hypoxic group and hypoxic preconditioning group was 8.6+/-2.0, 40.2+/-8.2, 26.4+/-6.1, respectively, and number of Caspase-3-positive cells of was 4.3+/ 1.2, 63.6+/-12.5, 45.7+/-9.8, respectively. The results showed that the expressions of Bcl-2 in both hypoxic group and hypoxic preconditioning group were significantly higher than that in the control group; and the expression of Bcl-2 in hypoxic preconditioning group was even higher than that in hypoxic group. The expressions of Caspase-3 in hypoxic group and hypoxic preconditioning group were also significantly higher than that in the control group; whereas the expression of Caspase-3 in hypoxic preconditioning group was significantly lower than that in hypoxic group. These results suggest that cortex cells are resistant to apoptosis via increased expression of Bcl-2 and lowered expression of Caspase-3 in the cortex and brain cells are thereby protected during hypoxic preconditioning. PMID- 18425315 TI - [Different effects of adenosine A2A receptors in the models of traumatic brain injury and peripheral tissue injury]. AB - Recently, activation of the adenosine A2A receptors has been shown to exert protection against peripheral tissue injuries but aggravation in the central nervous system (CNS) injuries. To explore the different effects of adenosine A2A receptors and try to perform some new treatment strategies for peripheral tissue and CNS traumas, we constructed the mouse models of skin trauma, skin combined radiation-impaired wound and traumatic brain injury (TBI), respectively. Wild type mice and A2A receptor gene knockout mice were both used in the experiments. In skin trauma and combined radiation-impaired wound models, the time of wound healing was observed, while in TBI model, neurological deficit scores, water content in injured brain and glutamate concentration in cerebral spinal fluid (CSF) were detected at 24 h after TBI. The results showed that in skin trauma and combined radiation-impaired wound models, CGS21680 (an agonist of the A2A receptors) promoted while A2A receptor gene knockout delayed the course of skin wound healing. On the contrary, in TBI model, A2A receptor gene knockout, not CGS21680, showed a protective role by inhibition of glutamate release. These data further indicate that promoting glutamate release may account for the different effects of A2A receptor activation in CNS injury and peripheral tissue injury models. These findings may provide some experimental evidence and a new strategy for clinical treatment of peripheral tissue damages by agonists of A2A receptors, while treatment of CNS injuries by antagonists of A2A receptors. PMID- 18425317 TI - Different glutamate receptor mechanisms in long-term depression induced by different stimulus patterns in the CA1 area of adult rat hippocampus. AB - Previous reports suggested that a novel stimulus pattern of multi-train stimulus at low-frequency (2-Hz or 5-Hz) could induce stable long-term depression (LTD) in the CA1 area of adult rat hippocampus. In the present study, in order to determine the mechanism in LTD induced by the two novel tetanus patterns, changes in the population spikes (PS) in the hippocampal CA1 area of adult rats following the multi-train stimulus in the presence of AP5 [antagonist of N-methyl-D aspartate receptors (NMDARs)] or MCPG [antagonist of type I/II metabotropic glutamate receptors (mGluRs)] were recorded. The results showed that both AP5 and MCPG inhibited the LTD induced by 2-Hz multi-train stimulus. The mean amplitude of population spikes (PSA) normalized to the baseline was (96.0+/-3.5)% after applying AP5 (n=10) and (95.7+/-4.1)% after applying MCPG (n=8), respectively, measured at 20 min post-tetanus. While 5-Hz multi-train tetanus failed to induce LTD in the presence of MCPG. The mean PSA was (73.6+/-4.4)% (n=10) and (98.2+/ 8.9)% (n=8) in the presence of AP5 and MCPG, respectively, measured at 35 min post-tetanus. So it is suggested that LTD induced by 2-Hz multi-train tetanus involves co-activation of NMDARs and mGluRs, while LTD induced by 5-Hz multi train tetanus is only related to activation of mGluRs. PMID- 18425316 TI - The selective 5-HT(1A) receptor antagonist WAY-100635 increases neuronal activity of the basolateral nucleus of the amygdala in 6-hydroxydopamine-lesioned rats. AB - In the present study, extracellular recording was used to examine the neuronal activity of the basolateral nucleus (BL) of the amygdala and the effects of systemic administration of the selective 5-HT(1A) receptor antagonist WAY-100635 on the neuronal activity in the normal rats and rats with 6-hydroxydopamine (6 OHDA)-produced lesions in the substantia nigra pars compacta (SNc). The results showed that the firing rates of BL projection neurons and interneurons were (0.39+/-0.04) Hz and (0.83+/-0.16) Hz in the normal rats, and (0.32+/-0.04) Hz and (0.53+/-0.12) Hz in 6-OHDA-lesioned rats. There was no significant difference in the firing rates of BL projection neurons and interneurons between the normal and 6-OHDA-lesioned rats. In the normal rats, all BL projection neurons fired in burst; 94% of BL interneurons fired in burst and 6% fired irregularly. In 6-OHDA lesioned rats, 85% of BL projection neurons displayed a burst firing pattern and 15% fired irregularly; 86% of BL interneurons had a burst firing pattern and 14% fired irregularly. The distribution of firing patterns of projection neurons and interneurons in the BL in 6-OHDA-lesioned rats did not differ from that in the normal rats. Systemic administration of WAY-100635 at 0.1 mg/kg body weight did not change the mean firing rates of projection neurons and interneurons in the BL in both normal and 6-OHDA-lesioned rats. However, a higher dose of WAY-100635 at 0.5 mg/kg body weight significantly decreased the mean firing rate of BL projection neurons from (0.43+/-0.07) to (0.15+/-0.02) Hz in the normal rats (P<0.01), but significantly increased the activity of BL projection neurons in 6 OHDA-lesioned rats from (0.37+/-0.08) to (0.69+/-0.18) Hz (P<0.004). The mean firing rates of BL interneurons in the normal and 6-OHDA-lesioned rats did not change after administration of a higher dose of WAY-100635 at 0.5 mg/kg body weight. These results demonstrate that the activity of BL neurons after substantia nigra dopaminergic lesion in the SNc is regulated by activation of intrinsic and extrinsic inputs, and that 5-HT(1A) receptors significantly contribute to the regulation of the activity of BL projection neurons in both normal and 6-OHDA-lesioned rats. Furthermore, WAY-100635 induced an increase in the mean firing rate of projection neurons in the BL in 6-OHDA-lesioned rats, suggesting that 5-HT(1A) receptor is likely to play a role in generating affective symptoms in Parkinson's disease. PMID- 18425318 TI - [Role of platelet-activating factor in progesterone synthesis and vascular endothelial growth factor expression in rat luteal cells]. AB - The present study aimed to investigate the role of platelet-activating factor (PAF) in progesterone synthesis and vascular endothelial growth factor (VEGF) expression in rat luteal cells. Immature (25-28 days old) female Sprague-Dawley rats were injected subcutaneously with 50 IU pregnant mare serum gonadotrophin (PMSG), and 25 IU human chorionic gonadotrophin (hCG) 48 h later, to induce follicular development and luteum formation. On day 6 after hCG administration (the day of hCG administration was the first day), the rats were killed by guillotine and the ovarian luteal cells were collected. After incubation for 24 h, luteal cells were incubated without or with different doses (0.1 MUg/mL, 1 MUg/mL, 10 MUg/mL) of PAF at 37 degrees C (5% CO(2)) for 24 h, and then progesterone concentration was evaluated by radioimmunoassay (RIA); apoptotic rate and VEGF mRNA expression in luteal cells were assessed by flow cytometry and RT-PCR, respectively. The results showed that PAF promoted progesterone production, with a maximal effect at 1 MUg/mL (P<0.05); PAF increased apoptotic rate but not in a dose-dependent manner, and 10 MUg/mL PAF enhanced apoptotic rate significantly (P<0.05); furthermore, PAF stimulated VEGF mRNA expression in luteal cells, especially at 1 MUg/mL (P<0.01). It is suggested that PAF regulates progesterone synthesis and VEGF mRNA expression in luteal cells to mediate corpus luteum formation in rat ovary. PMID- 18425320 TI - Analysis of high-frequency stimulation-evoked synaptic plasticity in mouse hippocampal CA1 region. AB - Extracellular recordings of field excitatory postsynaptic potential (fEPSP) is one of the most common ways for studies of synaptic plasticity, such as long-term potentiation (LTP) and paired-pulse plasticity (PPP). The measurement of the changes in the different components of fEPSP waveform, such as the initial slope, initial area, peak amplitude and whole area, were commonly used as criteria for the judgement of potentiation or depression of synaptic plasticity. However, the differences in the conclusions drawn from measuring different components of fEPSP waveform at the same recording have still been largely ignored. Here we compared high-frequency stimulation (HFS)-evoked synaptic plasticity, both LTP and PPP, by measuring different components of fEPSP waveform, including the initial slope, initial area, peak amplitude, whole area and time course. The results not only indicated the acceleration of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor kinetics underlies LTP in hippocampal CA1 region of mice, but also showed that different measurements of fEPSP waveform at the same recording result in different magnitudes of LTP and different forms of PPP in hippocampal CA1 region of mice. After HFS, the paired-pulse ratio was slightly decreased by measurement of the initial area, but obviously increased by measurement of the initial slope of the pair fEPSPs. These results might draw apparently contradictory conclusions. Therefore, careful and complete analysis of the data from different parts of fEPSP waveforms is important for reflection of the faithful changes in synaptic plasticity. PMID- 18425319 TI - Resveratrol inhibits electrical activity of paraventricular nucleus neurons in rat hypothalamic slices. AB - To study the role of resveratrol in the discharges of neurons in paraventricular nucleus (PVN) in hypothalamic slices, extracellular single-unit discharge recording technique was used. The effects of resveratrol were examined with glass microelectrodes in the rat PVN neurons at resting potential level. The results were as follows: (1) In response to the application of resveratrol (0.05, 0.5, 5.0 MUmol/L, n=29) to the superfusate for 2 min, the spontaneous discharge rate (SDR) of neurons in 28/29 (96.6%) hypothalamic slices significantly decreased in a dose-dependent manner; (2) Pretreatment with L-glutamate (0.2 mmol/L) led to a marked increase in the SDR in all 8/8 (100%) slices in an epileptiform pattern. The increased discharges were suppressed by the application of resveratrol (5.0 mmol/L) in all 8 slices; (3) In 8 slices, perfusion of the selective L-type calcium channel agonist, Bay K8644 (0.1 MUmol/L), induced a significant increase in the discharge rate in 8/8 (100%) slices. Resveratrol (5.0 MUmol/L) significantly attenuated the increased SDR in all 8 slices; (4) Pretreatment with the nitric oxide synthase (NOS) inhibitor N(omega)-nitro-L-arginine methyl ester (L-NAME, 50 MUmol/L) increased SDR in 7/8 (87.5%) slices, but did not affect the inhibitory effect of resveratrol (5.0 MUmol/L). These results suggest that resveratrol inhibits the electrical activity of PVN neurons and exerts neuroprotective actions on central neurons. The inhibitory effect of resveratrol is possibly related to the blockade of L-type calcium channel, but not due to NO release. PMID- 18425321 TI - [Electrophysiological characteristics of central neuronal dendrites and roles of dendritic back-propagating action potentials in modifications of synaptic plasticity]. AB - For expressing the condolences on the passing away of Dr. Hsiang-Tung Chang, one of the distinguished members of the Chinese Academia of Sciences, the pioneer studies on cortical dendritic potentials that Dr. Chang carried out in the 1950s and the prosperous progresses since then, especially, concerning the modifications of synaptic plasticity by the dendritic back-propagating action potentials were briefly reviewed. PMID- 18425322 TI - Genetic polymorphisms and head and neck cancer risk (Review). AB - The aim of this report is to review and evaluate, in a comprehensive manner, the published data regarding the contribution of genetic polymorphisms to risk of head and neck cancer (HNC). All relevant studies available in MEDLINE and published before July 2007 were identified. Studies carried out in humans that compared HNC patients with at least 1 standard control group were considered for analysis. Two hundred and eighteen publications and 3 published meta-analyses were identified. Seventy-five (34%) studies were conducted in Asian, 72 (33%) in American, and 68 (31%) in European countries. The most widely studied gene was GSTM1 (58 studies), followed by GSTT1 (42 studies), GSTP1 (codon 105, 22 studies) and p53 (codon 72, 20 studies). GSTM1, GSTT1, GSTP1, XRCC1 codons 194 and 399, and CYP1A1 codon 462 were examined by meta-analyses, and significant relations were found between the GSTM1-null genotype and an increased risk for HNC. In addition, increased risk for HNC was associated consistently with the ALDH2*1/*2, p53 codon 72 Pro/Pro and EPHX1 codon 113 Tyr/His and His/His genotypes. Cohort studies that simultaneously consider multiple genetic and environmental factors possibly involved in carcinogenesis of the head and neck are needed to ascertain not only the relative contribution of these factors to tumor development but also the contributions of their putative interactions. PMID- 18425323 TI - Characterization of gene expression regulated by American ginseng and ginsenoside Rg3 in human colorectal cancer cells. AB - American ginseng (Panax quinquefolius L., Araliaceae) possesses anti-cancer potential and is one of the most commonly used herbal medicines in the United States. Ginsenoside Rg3, one of the saponins in American ginseng, has been shown to inhibit tumor growth. In this study, we sought to characterize the downstream genes targeted by American ginseng extracts in HCT-116 human colorectal cancer cells. We first demonstrated that the content of Rg3 in American ginseng steamed at 120 degrees C for 2 h (referred to as S2h) was significantly increased when compared with that of the unsteamed ginseng. Both S2h and Rg3 exhibited antiproliferative effects on HCT-116 cells. Using the Affymetrix high density genechips containing more than 40,000 genes and ESTs, the gene expression profiling of HCT-116 cells were assayed. Microarray data indicated that the expression levels of 76 genes were changed significantly after treatment with S2h or Rg3, whereby it was found that 52 of the 76 genes were up-regulated while the remaining 24 were down-regulated. Ingenuity pathways analysis of top functions affected by both S2h and Rg3 were carried out. The most effected pathway is the Ephrin receptor pathway. To validate the microarray data, quantitative real-time PCR of six candidate target genes was conducted, whereby it was found that three genes were up-regulated (AKAPA8L, PMPCB and PDE5A) and three were down-regulated (PITPNA, DUS2L and RIC8A). Although further studies are needed to elucidate the mechanisms of action, our findings should expand the understanding of the molecular framework of American ginseng as an anti-cancer agent. PMID- 18425324 TI - HLA-A2 and -A24-restricted glypican-3-derived peptide vaccine induces specific CTLs: preclinical study using mice. AB - We previously reported that glypican-3 (GPC3) is uniquely overexpressed in human hepatocellular carcinoma and melanoma and that it is an ideal tumor antigen for immunotherapy in mouse models. We recently identified both HLA-A24 (A*2402) and H 2Kd-restricted GPC3298-306 (EYILSLEEL) and HLA-A2 (A*0201)-restricted GPC3144-152 (FVGEFFTDV), both of which can induce GPC3-reactive cytotoxic T cells (CTLs). The present study was a preclinical study in a mouse model that was conducted in order to design an optimal schedule for clinical trial of GPC3-derived peptide vaccine. When BALB/c mice were intradermally vaccinated at the base of the tail with Kd-restricted GPC3298-306 peptide mixed with incomplete Freund's adjuvant (IFA), the peptide-specific CTLs were induced. But the peptide alone could not induce peptide-specific CD8+ T cells. Furthermore, proteomic analyses showed that IFA protected the peptide against degradation in the human serum. Peptide reactive CTLs were induced by peptide vaccine in a dose-dependent manner. In addition, at least two vaccinations with a single dose >10 microg were needed for the induction of GPC3298-306-specific CTLs. But repeated vaccination with a lower dose of GPC3298-306 did not induce peptide-specific CTLs. Similarly, induction of an Ag-specific immune response by HLA-A2 GPC3144-152 depended on the dose administered. The results of this study suggested that IFA is one of the indispensable adjuvants for peptide-based immunotherapy, and that the immunological effect of peptide vaccines depends on the dose of peptide injected. PMID- 18425325 TI - A novel molecular mechanism for anticancer drug-induced ovarian failure: Irinotecan HCl, an anticancer topoisomerase I inhibitor, induces specific FasL expression in granulosa cells of large ovarian follicles to enhance follicular apoptosis. AB - Clinical use of CPT-11 combination chemotherapy frequently induces ovarian dysfunction in premenopausal and perimenopausal cancer patients, but its mechanism remains unclear. Mouse experiments were performed to clarify the molecular mechanism of CPT-11-induced ovarian dysfunction. Clinically therapeutic doses of CPT-11 were injected intraperitoneally into 8-week-old female MCH mice, and their ovaries were examined by the TUNEL assay to detect dead cells. Immunohistochemical examinations were simultaneously performed to detect the expression of activated caspase 3, Fas antigen and Fas ligand (FasL). Furthermore, normal murine ovarian tissue fragments were incubated with recombinant soluble FasL in organ cultures and stained by the TUNEL assay to detect apoptotic cells. Intraperitoneal CPT-11 injections induced specific TUNEL positive cells and cell death with cleaved caspase 3 expression among large ovarian follicular granulosa cells. Apoptotic follicles (follicles containing >/=10 TUNEL-positive cells per ovarian section) were only found among large follicles. The final apoptotic follicle ratios were approximately 30% of the total follicles independent of the CPT-11 dose, while CPT-11 dose-dependently enhanced apoptotic processes in murine ovarian follicles. Fas antigen was expressed in most ovarian cells, with extremely high expression levels detected in luteal cells. CPT-11 injections did not significantly increase the Fas expression levels in ovarian cells. Although no FasL expression was detected in normal ovarian tissues, CPT-11 injections significantly induced specific FasL expression in granulosa cells. Incubation of organ-cultured normal murine ovarian tissue fragments with recombinant mouse soluble FasL significantly increased the numbers of TUNEL-positive granulosa and luteal cells. In conclusion, CPT-11 dose dependently induced specific FasL expression in granulosa cells of developing ovarian follicles. The induced FasL reacted with the Fas antigen constitutively expressed on granulosa cells, such that apoptosis can only be enhanced and induced in granulosa cells in an autocrine and/or paracrine manner. This cell lineage-specific and differentiation stage-specific apoptosis in granulosa cells is thought to be the main molecular mechanism of the ovarian dysfunction induced by CPT-11 combination chemotherapy. PMID- 18425326 TI - Hyaluronan-mediated motility: a target in oral squamous cell carcinoma. AB - To characterize cancer-related gene expression changes in oral squamous cell carcinomas (OSCCs), we compared the gene expression profiles in OSCC-derived cell lines with human normal oral keratinocytes (HNOKs). Microarray analysis identified 166 genes that were up-regulated in OSCC-derived cell lines. Gene ontology analysis showed that cancer-related function had the highest significance. Among the genes mapped to the cancer-related network with the highest significance, the receptor for hyaluronan-mediated motility (RHAMM) was evaluated further for mRNA and protein expression in the OSCC cell lines, primary OSCCs. Overexpression of RHAMM protein was observed in all cell lines compared to HNOKs. Immunohistochemical analysis showed highly expressed RHAMM in primary OSCCs, whereas most corresponding normal tissues had no or significant down regulation of protein immunoreactivity. Real-time quantitative reverse transcriptase-polymerase chain reaction data agreed with the protein expression. Moreover, the RHAMM expression status was correlated with the TNM stage (P<0.001). The results suggested that RHAMM expression may be correlated with tumor aggressiveness and offer clues to the development of new treatments for human OSCCs. PMID- 18425327 TI - Pleiotropic effects of fenretinide in neuroblastoma cell lines and multicellular tumor spheroids. AB - The efficacy and mechanism of action of fenretinide (4-HPR), a vitamin A analogue, was investigated in a panel of six neuroblastoma cell lines and multicellular tumor spheroids. The latter are three dimensional cell aggregates and as such, a model for micrometastases. In all cell lines, the production of reactive oxygen species (ROS) increased with 163-680% after 1 h of treatment with 4-HPR. In addition, a decrease of the mitochondrial membrane potential of 30-75% was observed after 4 h of incubation with 4-HPR. A 6-12-fold difference was observed between the IC50 values for cell proliferation and viability between the most sensitive (IMR32) and most resistant (NASS) cell line towards 4-HPR. Flow cytometric analysis showed an increased amount of apoptotic bodies and no cell cycle arrest. The antioxidant Trolox completely inhibited the accumulation of 4HPR-induced ROS and prevented the 4HPR-associated cytotoxicity. In all neuroblastoma spheroids, 4-HPR induced a complete cytostasis at clinical relevant concentrations (3-10 microM). Immunohistochemical analysis of 4-HPR-treated spheroids showed a decreased staining for proliferation marker Ki-67 and an increased staining for cleaved-PARP, a marker of apoptosis. Our results suggest that 4-HPR might be a promising agent for the treatment of micrometastases and high-risk neuroblastoma. PMID- 18425328 TI - Detection of a hypersialylated beta1 integrin endogenously expressed in the human astrocytoma cell line A172. AB - Gliomas are the most common deadly brain tumors. Human cerebral tumors express high level of alpha5beta1 integrins. As a potential new target, alpha5beta1 was investigated here in two human astrocytoma cell lines, A172 and U87MG. We found that a hypersialylated beta1 integrin was endogenously expressed in A172 cells. It forms heterodimers with alpha5 subunits, localizes at the cell membrane and allows adhesion to fibronectin. This form of beta1 integrin was only recognized by the 9EG7 anti-beta1 antibody and appeared devoid of other specific antibody epitopes (12G10, TS2/16 and mAb13 shown here to be N-glycosylation sensitive). Overexpression of the beta1 integrin subunit in A172 cells not only increased the hypersialylated form but also led to the appearance of a non-hypersialylated beta1 form also addressed to the cell surface. Compared to wild-type A172 cells, beta1-A172 cells showed increased adhesion to fibronectin and decreased sensitivity to SJ749, a non-peptidic alpha5beta1 antagonist. In addition, beta1 A172 cells exhibited increased matrix dependence for normal cell cycling. Collectively, the data add new evidence for the role of beta1 glycosylation/sialylation in the regulation of integrin functions. PMID- 18425329 TI - Estrogen and antiestrogen-dependent regulation of breast cancer cell proliferation in multicellular spheroids: Influence of cell microenvironment. AB - Multicellular tumor spheroids, an in vitro 3-D model that simulates malignant cell contacts within a tumor, can be used to evaluate tumor response to therapeutic agents. We found that MELN (derived from MCF-7 cells) cells grown in 3-D as spheroids, remain highly sensitive to estradiol in terms of growth, down regulation of ERalpha expression and ERalpha-induced transcriptional activity. Estradiol induces cyclin D1 and CDK1 proteins in Ki-67 positive proliferating cells, whereas survivin is up-regulated in both Ki-67 positive proliferative outer layer of cells and around the necrotic zone in non-proliferating cells. OH Tam inhibits both estradiol-induced transcriptional activity and estradiol dependent growth of MELN spheroids. Consistent with its antiproliferative effect, we observed that OH-Tam induces an important decrease in the proportion of proliferating cells, positive for Ki-67, cyclin D1 and CDK1. But, in contrast to what was expected, OH-Tam treatment resulted in a decrease in the proportion of p21 positive cells. Furthermore, despite its ability to down-regulate survivin in MELN spheroids, OH-Tam did not trigger apoptosis. Taken together, these results indicate that this model, is more relevant to an in vivo situation than monolayer cultures. It could be useful to identify new markers of the response to endocrine treatment and to investigate the effects of drugs combination. PMID- 18425330 TI - Upregulation of Igf and Wnt signalling associated genes in pleomorphic adenomas of the salivary glands in PLAG1 transgenic mice. AB - The Pleomorphic adenoma gene 1 (PLAG1) is involved in various human neoplasias, including pleomorphic adenomas of the salivary glands. Moreover, the oncogenic role of PLAG1 was clearly demonstrated in two independent PLAG1 transgenic mouse founders, in which PLAG1 expression could be targeted to different tissues using the Cre/loxP system. MMTV-Cre-mediated targeted overexpression of PLAG1 in the salivary glands of double transgenic offspring mice, referred to as P1-MCre and P2-MCre mice, induced pleomorphic adenomas in this organ. Igf2, a genuine PLAG1 target gene, was highly upregulated in those tumours as well as in human pleomorphic adenomas of the salivary glands. These and previous observations in other PLAG1-induced tumours e.g. breast adenomyoepitheliomas emphasize the importance of Igf upregulation in such tumours. In this study, further evidence for the role of Igf2 in PLAG1-induced tumourigenesis, is reported. Inactivation of Igf2 in P1-MCre mice leads to a significant delay in tumour development. Since tumour development is not fully abrogated by inactivation of Igf2, other signalling pathways are likely to contribute to PLAG1-induced tumourigenesis as well. Further studies revealed that several genes such as H19, Dlk1, Gtl2, Igfbp2, Igfbp3 and genes involved in Wnt signalling, such as Wnt6, Cyclin D1 and beta-catenin are upregulated in P1-MCre mice in which Igf2 is inactivated. In conclusion, we clearly demonstrate upregulation of several genes associated with Igf and Wnt signalling in PLAG1-induced pleomorphic adenomas. Furthermore, inactivation of Igf2 does not affect upregulation of genes associated with Wnt signalling, which might suggest that both signalling pathways are involved. PMID- 18425331 TI - Antisense-mediated inhibition of survivin, hTERT and VEGF in bladder cancer cells in vitro and in vivo. AB - Since cancer cells are characterised by multiple genetic alterations the single inhibition of one tumour- associated gene might not be sufficient as a therapeutic strategy. We examined the effects of a combined inhibition of survivin, human telomerase reverse transcriptase (hTERT) and vascular endothelial growth factor (VEGF) with antisense oligodeoxynucleotides (AS-ODNs) and small interfering RNAs (siRNAs) in EJ28 and 5637 bladder cancer (BCa) cells. Following verification of the uptake of intraperitoneally applied fluorescence-labelled AS ODNs and siRNAs in subcutaneous BCa xenografts, the target-directed constructs were tested as single agents in SCID mice bearing subcutaneous EJ28. Simultaneous inhibition of two of the selected transcripts significantly enhanced cell viability reduction compared to the controls consisting of a target directed construct and an appropriate control construct without any homology to the human genome. The uptake of both antisense inhibitor types in the subcutaneous BCa was achieved even without a carrier. In vivo studies with 9 consecutive intraperitoneal injections with 20 mg/kg AS-ODNs or 4.6 mg/kg siRNAs revealed the biocompatibility of both antisense inhibitor types and showed anti-tumoural activity of the AS-ODNs used. PMID- 18425332 TI - Reduced T-cadherin expression and promoter methylation are associated with the development and progression of hepatocellular carcinoma. AB - Loss of T-cadherin expression has been reported in a number of human cancers. We previously reported that T-cadherin re-expression suppressed cell growth and motility in glioma. Here, we report that the T-cadherin expression was significantly decreased in human hepatocellular carcinoma (HCC) compared to adjacent normal liver. In addition, T-cadherin expression in HCC with metastasis was significantly lower than in HCC without metastasis. To determine the mechanism underlying the reduced T-cadherin expression in HCC, we examined T cadherin promoter methylation. We found that methylation of the T-cadherin promoter was present in 40% of HCC, but absent in all adjacent liver tissues. In the HCC with T-cadherin promoter methylation, the T-cadherin expression was significantly decreased compared to HCC without methylation. To provide a functional link between T-cadherin promoter methylation and T-cadherin growth regulation, we used the HepG2 hepatoma cell line that exhibits T-cadherin promoter methylation. Treatment of HepG2 cells with the demethylating agent 5-aza 2-deoxycytidine resulted in increased T-cadherin expression and reduced cell proliferation. These results demonstrate that the T-cadherin down-regulation by promoter methylation is associated with the development and progression of HCC, and suggest that T-cadherin is an important tumor suppressor in liver cancer. PMID- 18425333 TI - Silencing of ZNF217 gene influences the biological behavior of a human ovarian cancer cell line. AB - Zinc-finger protein 217 (ZNF217), a candidate oncogene on 20q13.2, can lead cultured human ovarian and mammary epithelial cells to immortalization, which indicates selective expression of ZNF217 affecting 20q13 amplification during critical early stages of cancer progression. In this study, we tested the hypothesis that ZNF217 is a key factor in regulating ovarian cancer proliferation and progression. We examined the effect of the inhibition of ZNF217 expression on proliferation and invasion by establishing the ZNF217 knockdown ovarian cancer cell line using RNA interference (RNAi). Our results showed that silencing of ZNF217 resulted in the effective inhibition of ovarian cancer cell growth and invasive ability. The results suggested that ZNF217 might play a crucial role in the proliferation and invasion of ovarian cancer. PMID- 18425334 TI - MMTV-cre-mediated fur inactivation concomitant with PLAG1 proto-oncogene activation delays salivary gland tumorigenesis in mice. AB - Proprotein convertases are serine endoproteases implicated in the proteolytic processing of a large variety of regulatory proteins. An important role of proprotein convertases in tumorigenic processes has been suggested by various studies. In this study, the role of the proprotein convertase furin in PLAG1 proto-oncogene-induced salivary gland tumorigenesis was investigated. PLAG1 overexpression in salivary glands has previously been shown to result in salivary gland tumors in 100% of mice within 5 weeks after birth. MMTV-cre-mediated inactivation of fur without over-expression of PLAG1 caused smaller but histologically normal salivary glands. Moreover, the lymph nodes close to the salivary glands were enlarged, and histology showed that they had activated follicles. When genetic ablation of 1 or 2 alleles of fur and overexpression of the PLAG1 transgene were simultaneously achieved, a significant delay in tumorigenesis was observed. Collectively, these results suggest an important role for furin in PLAG1-induced salivary gland tumorigenesis in mice. PMID- 18425335 TI - Evaluation of aqueous extracts of Taraxacum officinale on growth and invasion of breast and prostate cancer cells. AB - Ethnotraditional use of plant-derived natural products plays a significant role in the discovery and development of potential medicinal agents. Plants of the genus Taraxacum, commonly known as dandelions, have a history of use in Chinese, Arabian and Native American traditional medicine, to treat a variety of diseases including cancer. To date, however, very few studies have been reported on the anti-carcinogenic activity of Taraxacum officinale (TO). In the present study, three aqueous extracts were prepared from the mature leaves, flowers and roots, and investigated on tumor progression related processes such as proliferation and invasion. Our results show that the crude extract of dandelion leaf (DLE) decreased the growth of MCF-7/AZ breast cancer cells in an ERK-dependent manner, whereas the aqueous extracts of dandelion flower (DFE) and root (DRE) had no effect on the growth of either cell line. Furthermore, DRE was found to block invasion of MCF-7/AZ breast cancer cells while DLE blocked the invasion of LNCaP prostate cancer cells, into collagen type I. Inhibition of invasion was further evidenced by decreased phosphorylation levels of FAK and src as well as reduced activities of matrix metalloproteinases, MMP-2 and MMP-9. This study provides new scientific data on TO and suggests that TO extracts or individual components present in the extracts may be of value as novel anti-cancer agents. PMID- 18425336 TI - ERCC1 codon 118 polymorphism is a useful prognostic marker in patients with pancreatic cancer treated with platinum-based chemotherapy. AB - Excision repair cross-complementation 1 (ERCC1) has been reported to play a major role in the response to platinum-based therapies. It has recently been proposed that a synonymous polymorphism at codon 118 converting a common codon usage (AAC) to an infrequent one (AAT) may impair ERCC1 translation and to affect the response to cisplatin chemotherapy. We analyzed the association between this polymorphism and clinical outcome in 67 pancreatic cancer patients treated with cisplatin and S-1 or with S-1 alone. ERCC1 codon 118 polymorphism was analyzed using PCR-RFLP. Thirty-nine of the patients (58.2%) were homozygous for AAC codon, 7 (10.4%) were homozygous for AAT codon, and 21 (31.3%) were heterozygous. Among those treated with cisplatin and S-1, no significant difference in objective response rate was observed between genotypes. However, the patients with one or two AAT codons had a significantly longer progression-free survival (PFS) and overall survival (OS) than those homozygous for AAC allele (PFS: 338 days vs 106 days, p=0.006, OS: 763 days vs 415 days, p=0.030). In contrast, no significant difference in PFS or OS was observed between genotypes among the patients treated with S-1 alone. ERCC1 polymorphism may be a useful prognostic marker in patients with pancreatic cancer treated with platinum-based chemotherapy. PMID- 18425337 TI - Meta-analysis of long-term mobile phone use and the association with brain tumours. AB - We evaluated long-term use of mobile phones and the risk for brain tumours in case-control studies published so far on this issue. We identified ten studies on glioma and meta-analysis yielded OR = 0.9, 95% CI = 0.8-1.1. Latency period of > or =10-years gave OR = 1.2, 95% CI = 0.8-1.9 based on six studies, for ipsilateral use (same side as tumour) OR = 2.0, 95% CI = 1.2-3.4 (four studies), but contralateral use did not increase the risk significantly, OR = 1.1, 95% CI = 0.6-2.0. Meta-analysis of nine studies on acoustic neuroma gave OR = 0.9, 95% CI = 0.7-1.1 increasing to OR = 1.3, 95% CI = 0.6-2.8 using > or =10-years latency period (four studies). Ipsilateral use gave OR = 2.4, 95% CI = 1.1-5.3 and contra lateral OR = 1.2, 95% CI = 0.7-2.2 in the > or =10-years latency period group (three studies). Seven studies gave results for meningioma yielding overall OR = 0.8, 95% CI = 0.7-0.99. Using > or =10-years latency period OR = 1.3, 95% CI = 0.9-1.8 was calculated (four studies) increasing to OR = 1.7, 95% CI = 0.99-3.1 for ipsilateral use and OR = 1.0, 95% CI = 0.3-3.1 for contralateral use (two studies). We conclude that this meta-analysis gave a consistent pattern of an association between mobile phone use and ipsilateral glioma and acoustic neuroma using > or =10-years latency period. PMID- 18425338 TI - CDK inhibitor enhances the sensitivity to 5-fluorouracil in colorectal cancer cells. AB - Thymidylate synthase (TS) is a dNTP synthetic enzyme and is also a target enzyme of 5-fluorouracil (5-FU). 5-FU is one of the anticancer agents most frequently used for the treatment of colorectal cancers. However, the clinical rate of response to its use as a single agent is not exceptionally high. Therefore, various combination chemotherapies have been devised. The elevated expression of TS in cancer cells is a serious obstacle in the clinical use of 5-FU. In the present study, TS expression was up-regulated by the knockout of the p21WAF1/CIP1 gene in human colorectal cancer HCT116 cells, suggesting that TS expression is mediated through the inhibition of cyclin-dependent kinase (CDK). Based on these findings, we tested whether the CDK inhibitor (CDKI) SU9516, acted as a suppressor of TS. SU9516 effectively reduced the expression of TS in a dose dependent manner. Furthermore, the reduction of TS expression resulted in enhancement of the sensitivity to 5-FU in human colon cancer DLD-1 cells. Thus, SU9516 might be a promising compound for combination chemotherapy with 5-FU. PMID- 18425339 TI - Prophylactic pelvic irradiation as part of primary therapy in uterine sarcomas. AB - In a complete geographic series of 155 cases of primary uterine sarcomas, prophylactic pelvic irradiation was used as standard postoperative therapy in the majority of the cases. Vaginal brachytherapy was also added as a boost. The histology was leiomyosarcoma in 62 cases (40%), carcinosarcoma in 60 (39%), endometrial stromal sarcoma in 25 (16%), and other types in 8 cases (5%). The primary surgery was extended hysterectomy in 11 cases (7%), simple hysterectomy in 110 (71%), and supravaginal hysterectomy in 12 (8%). In 22 cases (14%) no major surgery was possible. In the complete series, 62 recurrences (40%) were recorded. Local (9%), regional (9%), and distant recurrences (28%) were the most frequent. The type of surgery was associated with the risk of tumor recurrence. Extended surgery reduced the risk of local and regional recurrences. The 5-year overall survival rate was 42% and the recurrence-free survival (RFS) was 37%. The number of mitoses was significantly (P=0.007) associated with survival. The locoregional RFS rate was 75% for patients treated with adjuvant irradiation and 83% for patients treated with primary surgery alone. Serious late tissue reactions from the bladder and intestine occurred in 7% of the irradiated cases. The locoregional tumor control rate was high in this series of patients, but no significant difference was found between patients treated with surgery alone and surgery plus postoperative pelvic irradiation. This was true for all histological subtypes of the uterine sarcomas. However, this was not a randomized study and selection bias cannot be ruled out. PMID- 18425340 TI - Evaluation of a nanotechnology-based carrier for delivery of curcumin in prostate cancer cells. AB - We have initiated studies to enhance targeted delivery of an anticancer agent, curcumin, for prostate cancer treatment by incorporating this agent into the liposomes (nanodelivery vehicles primarily composed of phospholipids) coated with prostate membrane specific antigen specific antibodies. We prepared curcumin loaded liposomes of various lipid compositions by sonication at an average size of 100-150 nm. Un-entrapped curcumin was removed by size exclusion chromatography. Data show that curcumin preferentially partitioned into liposomes prepared from dimyristoyl phosphatidyl choline (DMPC) and cholesterol among the various compositions tested. The anti-proliferative activity of liposomal curcumin was studied using two human prostate cancer cell lines (LNCaP and C4-2B) by a tetrazolium dye-based (MTT) assay. Treatment of cells with liposomal curcumin (5-10 microM) for 24-48 h at 37 degrees C resulted in at least 70-80% inhibition of cellular proliferation without affecting their viability. On the other hand, free curcumin exhibited similar inhibition only at 10-fold higher doses (>50 microM). We also observed that LNCaP cells were relatively more sensitive to liposomal curcumin mediated block of cellular proliferation than C4 2B cells. We are currently developing liposome formulations with targeting ability to further improve the efficacy of curcumin in vivo. PMID- 18425341 TI - Migration of renal carcinoma cells is dependent on protein kinase Cdelta via beta1 integrin and focal adhesion kinase. AB - Migration and adhesion of tumor cells are essential prerequisites for the formation of metastases in malignant diseases. Protein kinase C (PKC) has been shown to regulate cell migration, adhesion and proliferation. In order to identify a connection between PKC isoforms and tumor progression in renal cell carcinoma (RCC), the influence of PKC isoforms on cell migration, adhesion and proliferation and possible influences of the activity of integrins and focal adhesion kinase (FAK) were analyzed in RCC cells. The experiments were performed in the RCC cell line CCF-RC1 after pre-incubation of the cells with the PKC inhibitors GF109203X, GO6976, RO31-8220 and rottlerin. Cell migration and adhesion were assessed through chemotaxis analysis and adhesion to an endothelial monolayer, respectively. Cell proliferation was analysed by a BrdU incorporation assay. The expression and activity of beta1 integrins and FAK were analysed by Western blot analysis. GF109203X reduced cell migration to 69%, the activity of beta1 integrins to 63% and FAK expression to 82% compared to untreated cells. Rottlerin reduced cell migration in a concentration-dependent manner to 36%, cell proliferation to 81%, expression and activity of beta1 integrins to 72 and 79%, and expression and activity of FAK to 56 and 76% of untreated cells, respectively. RO31-8220 also reduced the expression and activity of beta1 integrins as well as the expression of FAK to 84, 66 and 66% of untreated cells, respectively. GO6976 reduced the expression of FAK to 60% of untreated cells. Cell migration was only slightly reduced by GO6976 to 84% of untreated cells, and cell adhesion remained uninfluenced. These findings show a critical role of PKCdelta in the regulation of tumor cell migration, which seems to be caused by affecting the expression and activity of beta1 integrins and FAK. These results can provide a basis for new strategies in preventing metastases of renal cell carcinoma. PMID- 18425342 TI - Constitutive activation of p70 S6 kinase is associated with intrinsic resistance to cisplatin. AB - Cisplatin is widely used for the treatment of solid tumors, including small cell lung cancers, but its success is often compromised due to relapse and resistance to further treatment. p70 ribosomal S6 kinase (p70S6K) has been shown to be upregulated in lung cancer cells. In the present study, we investigated whether the p70S6K pathway contributes to cisplatin resistance in human small cell lung cancer H69 cells. The levels of phosphorylated p70S6K and its downstream target S6 but not total p70S6K or S6 were elevated in the H69 cells that acquired resistance to cisplatin (H69/CP) compared to parental H69 cells. Cisplatin treatment resulted in the activation of p70S6K and downregulation of p70S6K was associated with cisplatin-induced PARP cleavage. While the ability of cisplatin to induce apoptosis was attenuated in H69/CP cells, inhibition of p70S6K by rapamycin enhanced cisplatin-induced apoptosis in these cells as evident by the increase in cisplatin-induced poly(ADP-ribose) polymerase (PARP) cleavage. The phosphoinositide 3-kinase (PI3K) inhibitor Ly294002 alone induced PARP cleavage and further augmented cisplatin-induced PARP cleavage. In contrast, inhibition of extracellular signal-regulated kinase (ERK) by U0126 attenuated cisplatin-induced PARP cleavage. Both rapamycin and Ly294002 enhanced cisplatin-induced acti-vation of ERK1/2. Taken together, these results suggest that activation of p70S6K contributes to cisplatin resistance in small cell lung cancer H69 cells, and inhibition/downregulation of p70S6K as well as activation of ERK1/2 could circumvent cisplatin resistance. PMID- 18425343 TI - Prevention of amyloid beta-induced memory impairment by fluvastatin, associated with the decrease in amyloid beta accumulation and oxidative stress in amyloid beta injection mouse model. AB - Alzheimer's disease (AD), the most common cause of dementia in the elderly, is characterized by amyloid beta (Abeta)-containing plaques and neurofibrillary tangles, and synaptic and neuronal loss, along with progressive cognitive impairment. Although growing evidence suggests the beneficial effects of 3 hydroxy-3-methylglutaryl coenzyme A reductase inhibitors (statins) on AD, this notion is still controversial. To evaluate the efficacy of statins for Abeta induced cognitive impairment, we employed an Abeta injection model. Using this model, the present study demonstrated that pretreatment with fluvastatin, but not post-treatment just after Abeta exposure, prevented Abeta-induced memory impairment. We also observed that fluvastatin significantly decreased Abeta accumulation and oxidative stress after Abeta injection. Mice treated with simvastatin, but not fluvastatin, did not demonstrate the prevention of Abeta induced memory impairment, and showed no significant decrease in oxidative stress. More importantly, fluvastatin significantly prevented the loss of neurons in the basal forebrain induced by Abeta. Overall, the present study demonstrated that fluvastatin significantly prevented memory impairment induced by Abeta. The beneficial effects of fluvastatin might be explained by the preservation of neurons through a significant decrease in Abeta accumulation and oxidative stress. In clinical practice, the timing of the start of fluvastatin treatment might be critical in achieving a beneficial effect on cognitive function. PMID- 18425344 TI - Osteogenic potential of embryonic stem cells in tooth sockets. AB - Embryonic stem cells (ESCs) are established from blastocysts and give rise to various types of cells and tissues. In the present study, we assessed the osteogenic potential of ESCs using in vitro culture conditions and in vivo differentiation in tooth sockets. An ESC-derived embryoid body (EB) was formed and subsequently induced to an osteogenic lineage. The differentiated EB cells exhibited increased expression of various osteogenic markers as determined by real-time PCR analysis. Likewise, the differentiated EB-derived cells had enhanced alkaline phosphatase activity and calcium accumulation, as determined by cytochemical methods. For in vivo transplantation, mixtures of ESCs and hydroxyapatite/ tricalcium phosphate particles or EBs alone were transplanted into female rat tooth sockets. After 12 weeks, we observed formation of osteogenic structure in the tooth sockets without evidence of teratomas. These data suggest that pluripotent ESCs can serve as an alternative source for the reconstruction of craniofacial structures, as well as for further applications. PMID- 18425345 TI - An immunohistochemical, histological, and electron-microscopic study of the human periodontal ligament during orthodontic treatment. AB - The periodontal ligament lies between the hard tissues of alveolar bone and cementum of teeth and serves to anchor the tooth to the alveolus and functions as a cushion between these hard tissues to migrate occlusal force during mastication. This tissue is always exposed to mechanical stress during mastication. When occlusal forces exceed the adaptive capacity of the periodontal ligament, the periodontal ligament tissue will be injured and then occlusal trauma will occur. The different modifications of periodontal ligament during load deformation can be monitored by analysis of the expression of different collagen types and fibronectin, with immunohistochemical techniques, and by morphological study of ligament, with light- and transmission electron microscopic techniques. The use of continued and light orthodontic force generates a pressure of ligament with ejection of parodontal fluid externally and partial closing of vessels. On these basis we performed a study in order to evaluate periodontal ligament collagen types I and IV and the fibronectin modifications induced by application of a precalibrated orthodontic strength. We integrated these results, with light and transmission electron-microscopic observations, in order to evaluate the morphological modifications of periodontal tissue. Our observations showed that the type I collagen immunofluorescence staining is increased in the pressure side; in the tension side, it shows prior to treatment an increase, and after 72 h of treatment, a diminution of the staining pattern. Type IV collagen staining is reduced in both sides, but increased gradually after 7 days from treatment; finally, fibronectin staining pattern is gradually increased in the pressure side and reduced in the tension side. In light and transmission electron-microscopic observations it is possible to show a reduction of vessels at 72 h from treatment, and an increase of vessels after 7 days from treatment. The Malassez's epithelial residues are decreased at 72 h, while they are increased after 7 days from treatment. The modifications of immunofluorescence staining patterns of tested proteins revealed angiogenesis and reparative processes, and a thickening of fibrillar matrix as a defensive reply to mechanical stress. The modification of normal staining patterns of tested protein in our observations, could be determined by variation of scaffold geometry of periodontal ligament. The reduced movements of contraction and relaxation of periodontal ligament, due to orthodontic treatment, provoke a loss of mechanical stresses transmitted over ligament surface. Mechanical signals, therefore, could be integrated with other environmental signals and transduced into biochemical signals through force-dependent changes in scaffold geometry. Physical forces of gravity, hemodynamic stresses, and movement play a critical role in tissues, since the cells use tensegrity architecture for their structural organization. PMID- 18425346 TI - Molecular detection of circulating tyrosinase mRNA: optimization in a preclinical xenograft mouse melanoma model and further evaluation in samples from advanced melanoma patients. AB - We designed high-affinity primers for the mRNA sequence of human tyrosinase to test the value of molecular detection of circulating melanoma cells by reverse transcription-polymerase chain reaction (RT-PCR). The optimization process included in vitro settings and in vivo studies in a xenograft mouse model. We detected tyrosinase expression with at least 40 pg and 1.5 pg of total RNA extracted from cultured SKmel human melanoma cells, using a first round of PCR amplification and nested PCR, respectively. Human tyrosinase expression was found in the blood of nude mice bearing subcutaneous SKmel tumors, and the expression bands were stronger after manipulation of the tumor mass. We also examined the fate of circulating melanoma cells in the present melanoma model. Tyrosinase expression declined in blood 6 h after a direct intravenous injection of SKmel cells. A preliminary study in human blood samples demonstrated a baseline positive tyrosinase determination in 64% (16/25) of advanced melanoma patients using the RT-PCR nested assay. Baseline tyrosinase expression was significantly associated with disease progression after 12 months, and sequential determination during follow-up of the remaining disease-free patients showed a progressive increase of negative results. PMID- 18425347 TI - BigH3 protein expression as a marker for breast cancer. AB - The current hypothesis of tumorigenesis in humans suggests that cancer cells acquire their hallmarks of malignancy through the accumulation of advantageous gene activation and inactivation events over long periods of time. For breast cancer development, this multistep process may manifest itself as a sequence of pathologically defined stages. It is widely held that breast cancer originates at the premalignant stage of atypical ductal hyperplasia, progresses to the preinvasive stage of ductal carcinoma in situ, and culminates in the potentially lethal stage of invasive ductal carcinoma. Tumor grade has been a highly valuable prognostic factor for breast cancer, and high-grade ductal carcinoma in situ lesions are associated with poor clinical outcome. The aim of this work was to investigate the BigH3 protein expression changes associated with various stages of breast cancer progression in comparison to benign specimens using tissue microarray technology. Pathological characteristics of breast tissues ranged from benign lesions to breast cancers either of lobular or ductal carcinomas in origin, and included in situ ductal carcinomas, lobular carcinomas, infiltrating ductal carcinomas, carcinomas, scirrhous carcinomas, adenocarcinomas and infiltrating colloid carcinomas. BigH3 protein expression was analyzed by immunohistochemistry in 192 cases of breast tumors. Results indicated a decrease in BigH3 protein expression from benign tissues to in situ ductal carcinoma, lobular carcinoma, infiltrating ductal carcinomas, carcinomas, scirrhous carcinoma, adenocarcinomas to infiltrating colloid carcinomas. We observed that the benign tissue had a 23-fold increase in BigH3 protein expression compared to the infiltrating colloid carcinoma which was the most malignant tissue analyzed. In summary, these studies confirmed the suppressor effect of the BigH3 gene expressed as protein expression in those processes related to the progression of breast tumorigenesis. We conclude that this protein can be used as a marker for breast cancer progression. PMID- 18425348 TI - Immunoglobulin heavy chain gene analysis in bone marrow biopsies and corresponding lymph node specimens: dependency on pre-treatment, histological subtype and extension of B-cell lymphoma. AB - Bone marrow biopsies (BMB) are the conventional staging method for assessing marrow involvement by lymphoma. Morphological criteria provide basic data determining their dignity, but concerning microfocal infiltrates, these criteria are rather inaccurate. Here, by examination of immunoglobulin H (IgH) receptor rearrangement and comparison of medullar and nodular lymphoma sites, diagnostic reliability was improved. Employing non-nested IgH rearrangement analysis with FR3A and JHa consensus primers, B-cell clonality was assessed on glutardialdehyde fixed, decalcified BMB with and without lymphoma infiltration and on the corresponding lymph node specimens. Malignancy was confirmed when polymerase chain reaction (PCR) generated no more than two peaks and was observed in 60% of the medullar B-cell lymphoma. Comparison of lymph node tissues and BMB revealed an identical pattern in 50% of the probes. In 25% of the cases a single clonal peak derived from the lymph node tissues was also observed in the BMB but was surrounded by additional peaks. Here, direct comparison of the data permitted determination of lymphoma in the BMB. Therefore, IgH FR3 PCR analysis is a suitable tool to examine small lymphoid infiltrates in BMB, and direct comparison with corresponding nodal lymphoma can further facilitate estimation of their dignity. PMID- 18425349 TI - Ganoderic acids suppress growth and invasive behavior of breast cancer cells by modulating AP-1 and NF-kappaB signaling. AB - Structurally related lanostane-type triterpenes, ganoderic acid A, F and H (GA-A, GA-F, GA-H), were identified in an oriental medicinal mushroom Ganoderma lucidum. In the present study we evaluated the effect of GA-A, GA-H and GA-F on highly invasive human breast cancer cells. We showed that GA-A and GA-H suppressed growth (cell proliferation and colony formation) and invasive behavior (adhesion, migration and invasion) of MDA-MB-231 cells. Our results suggest that GA-A and GA H mediate their biological effects through the inhibition of transcription factors AP-1 and NF-kappaB, resulting in the down-regulation of expression of Cdk4 and the suppression of secretion of uPA, respectively. Furthermore, the activity of ganoderic acids is linked to the hydroxylation in the position 7 and 15 (GA-A) and 3 (GA-H) in their triterpene lanostane structure. In conclusion, hydroxylated triterpenes from G. lucidum could be promising natural agents for the therapy of invasive breast cancers. PMID- 18425350 TI - Thermosensitization by parthenolide in human lung adenocarcinoma A549 cells and p53- and hsp72-independent apoptosis induction via the nuclear factor-kappaB signal pathway. AB - The thermo-enhancement effects of the sesquiterpene lactone parthenolide (PTL), which targets the transcription factor nuclear factor-kappaB (NF-kappaB), and hyperthermia at 40, 42 and 44 degrees C on the human lung adenocarcinoma A549 cell line were investigated in vitro. Thermotherapy using a combined treatment with PTL (0.02 microM) prior to hyperthermia showed synergistic thermo enhancement effects towards A549 cells. The expression of p53 and hsp72 proteins following the application of PTL and hyperthermia at 44 degrees C, both alone and in combination, were examined to investigate whether p53 and hsp72 participated in apoptosis induction via the NF-kappaB signal pathway. After treatment with PTL alone, Hsp72 was only slightly induced, which was the same as for the control, while the level following the combination treatment was not significantly different as compared with hyperthermia alone. In addition, the level of p53 after the combination treatment was only slightly increased in comparison with hyperthermia alone. The kinetics of apoptosis and necrosis induction during the incubation periods following PTL exposure and hyperthermia, and the combination of both were also determined. The incidence of apoptosis following hyperthermia alone was approximately 0.6% on average after 12, 24 and 48 h of incubation, while that of PTL alone was approximately 1.7%, and that with the combination treatment was around 2.3%. Thus, induction of apoptosis following the combination treatment was increased as compared to each treatment alone. With regard to the kinetics of necrosis, the incidence of necrosis after treatment with hyperthermia alone was approximately 2.7%, while that with the combination treatment was lower, at around 2.2%. We hypothesized that cells treated with PTL had an altered arrangement of stressed cells undergoing the transformation from necrotic cell death to apoptotic cell death via another mechanism. Our results suggested that the PTL-induced apoptosis of A549 cells was due to the direct suppression of NF kappaB activity in a p53- and hsp72-independent manner based on NF-kappaB signaling. PMID- 18425351 TI - Steroidogenic acute regulatory protein gene expression, steroid-hormone secretion and proliferative activity of adrenocortical cells in the presence of proteasome inhibitors: in vivo studies on the regenerating rat adrenal cortex. AB - Previous studies have shown that proteasome inhibitors promote the accumulation of steroidogenic acute regulatory protein (StAR) in cultured rat adrenocortical cells. Unexpectedly, this response was associated with a moderate lowering in the corticosterone secretion and proliferation rate of cultured cells. Hence, we studied the effects of proteasome inhibitors MG115 and MG132 on the secretion and proliferative activity of the regenerating adrenal cortex in rats 5 days after surgery. Animals were given two subcutaneous injections of 0.15 or 1.5 nmol/100 g of inhibitors 24 and 12 h before decapitation. Real-time PCR and Western blotting showed that StAR expression, both mRNA and protein, was markedly lower in regenerating adrenals than in the intact gland of sham-operated rats. Neither MG115 nor MG132 affected StAR expression in regenerating gland. Inhibitors induced a slight decrease in the plasma concentrations of aldosterone and corticosterone, but did not significantly alter metaphase index of the regenerating adrenal cortex. Our findings provide the first evidence that down regulation of StAR occurs during the early stages of adrenal regeneration. Moreover, this suggests that the steroidogenic pathway is more sensitive to proteasome inhibitors than that regulating proliferative activity of regenerating adrenal cortex in the rat. PMID- 18425352 TI - Protection of telomeres 1 protein levels are associated with telomere length in gastric cancer. AB - Protection of telomeres 1 (Pot1) is a telomere-associated protein, which binds to the single-stranded DNA extensions of telomeres and regulates telomere length. Pot1 production was examined and compared with telomere length in gastric cancer. Pot1 production and telomere lengths were assessed in 5 human gastric cancer cell lines by immunoblotting and Southern blotting, respectively. Pot1 intracellular localization was examined with protein fractionation. Pot1 index and telomere volume were examined in human gastric mucosa and cancer by immunohistochemistry and in situ hybridization. Pot1 protein levels, which were lower than those in the lymphocytes of healthy persons, were significantly correlated with telomere length in gastric cancer cells (P=0.0167). Pot1 protein was mainly detected in the nuclear fraction and increased by G2/M blocking with nocodazole in MKN28 cells. Pot1 indexes were correlated with telomere volumes in gastric cancers (P<0.0001). Pot1 index was decreased in gastric epithelia distant from cancer (84+/-14%), in peritumoral epithelia (72+/-24%), and in stage I-II (39+/-14%) and stage III-IV (23+/-14%) gastric cancers (P<0.0001). Pot1 index was lower in stage III-IV than in stage I-II gastric cancers (P<0.05). Pot1-low cases showed advanced cancer invasion (P<0.05). Thus, Pot1 production was closely associated with telomere length in gastric mucosa and cancers. Pot1 might be a good in situ marker for the examination of cell-specific telomere length. PMID- 18425353 TI - Nuclear localization of regucalcin is enhanced in culture with protein kinase C activation in cloned normal rat kidney proximal tubular epithelial NRK52E cells. AB - In this study we investigated whether the nuclear localization of regucalcin in cloned normal rat kidney tubular epithelial NRK52E cells is regulated after culture with hormonal signaling factors. Stable regucalcin/pCXN2 transfectants with subconfluent monolayers were further cultured for 24 or 48 h in a serum-free medium containing either vehicle, tumor necrosis factor-alpha (TNF-alpha), transforming growth factor-beta1 (TGF-beta1), parathyroid hormone (PTH), phorbol 12-myristate 13-acetate (PMA), or other factors. Culture with TNF-alpha (1.0 ng/ml of medium) or TGF-beta1 (5.0 ng/ml) for 48 h caused a significant decrease in regucalcin mRNA levels in NRK52E cells (wild-type), while regucalcin mRNA levels were markedly increased in the presence of PMA (10(-6) M), an activator of protein kinase C, in wild-type cells. Immunocytochemical observation showed that HA-regucalcin was markedly localized in the nucleus of HA-regucalcin/ phCMV2 transfected cells. The nuclear localization was enhanced in culture with BS (5%), PTH (10(-7) M), Bay K 8644 (2.5x10(-6) M), or PMA (10(-6) M) for 24 or 48 h. Culture with staurosporine, an inhibitor of protein kinase C, caused a remarkable decrease in the localization of HA-regucalcin in the nucleus of HA-RGPR p117/phCMV2-transfected cells with PMA. Culture with PMA (10(-6) M) for 24 or 48 h caused a remarkable increase in nuclear regucalcin protein levels. The effect of PMA in increasing nuclear regucalcin levels was completely absent in culture with staurosporine (10(-8) M). The nuclear localization of regucalcin in the stable regucalcin/pCXN2-transfected cells (transfectant) increased markedly as compared with that of wild-type cells, whereas the increase was less evident in the transfectants cultured with staurosporine. This study demonstrated that regucalcin localizes in the nucleus of cloned normal rat kidney proximal tubular epithelial NRK52E cells, and that its nuclear localization is enhanced through an intracellular signaling process which involves protein kinase C. PMID- 18425354 TI - Autocrine stimulation of human hepatocytes triggers late DNA synthesis and stabilizes long-term differentiation in vitro. AB - Isolated human hepatocytes are of great value in investigating cell transplantation, liver physiology, pathology, and drug metabolism. Though hepatocytes possess a tremendous proliferative capacity in vivo, their ability to grow in culture is severely limited. We postulated that repeated medium change, common to most in vitro systems, may prevent long-term maintenance of hepato specific functions and growth capacity. To verify our hypotheses we compared the DNA synthesis and differentiation status of isolated human hepatocytes, cultured in medium which was renewed every day or was not changed for 3 weeks ('autocrine' setting). Daily medium change led to rapid hepatocellular de-differentiation without any signs of DNA replication. In contrast, the autocrine setting allowed hepatocytes to become highly differentiated, demonstrated by an elevated ASGPr expression level, and increased albumin and fibrinogen synthesis and release. Cytokeratin 18 filaments were stably expressed, whereas cytokeratin 19 remained undetectable. Hepatocytes growing in an autocrine fashion were activated in the presence of hepatocyte growth factor (HGF), evidenced by c-Met phosphorylation. However, HGF response was not achieved when the culture medium was renewed daily. Furthermore, the autocrine setting evoked a late but strong interleukin 6 release into the culture supernatant, reaching maximum values after a 10-day cultivation period, and intense BrdU incorporation after a further 5-day period. Our data suggest that preservation of the same medium creates environmental conditions which allow hepatocytes to control their differentiation status and DNA synthesis in an autocrine fashion. Further studies are necessary to identify the key mediators involved in autocrine communication and to design the optimal culture configuration for clinical application. PMID- 18425355 TI - Pseudomonas fluorescens proliferates in a mouse organ homogenate at low temperature. AB - In this study we observed the proliferation of Pseudomonas fluorescens (P. fluorescens) in mouse organ homogenates at 4 degrees C. P. fluorescens secreted a protease possessing properties different from those of the mammalian tissue proteases. The specificity of this protease required a basic amino acid residue at the P1 position at a pH optimum of 6.0. The specificity of the protease was similar to that of trypsin, but the pH optimum was different. The protease mildly degraded elastin-Congo red; this suggests that the protease serves as an alternative for elastase in the case of P. fluorescens strains that lack virulent elastase. The protease was identified as an alkaline protease of P. fluorescens by liquid chromatography-tandem mass spectrometry analysis. Our results show that proteome analysis of the soluble proteins is useful in identifying bacterial species, particularly the bacterial contaminants in samples containing antibiotics. PMID- 18425356 TI - Gene expression profiling of breast cells induced by X-rays and heavy ions. AB - Several genetic aberrations and gene expression changes have been shown to occur when cells are exposed to various types of radiation. The integrity of DNA depends upon several processes that include DNA damage recognition and repair, replication, transcription and cell cycle regulation. Ionizing radiation has many sources, including radon decay from the soil and X-rays from medical practice. Epidemiological evidence indicates a risk for cancer by inducing genetic alterations through DNA damage, and molecular alterations have been reported in epidemiological studies of the A-bomb survivors. A spontaneously immortalized human breast epithelial cell model, MCF-10F, was used to examine the gene expression profiling of breast cells induced by X-ray and heavy ion exposure, by a cDNA expression array of DNA damage and repair genes. This cell line was exposed to 10, 50, 100 and 200 cGy of either X-rays or heavy ions and gene expression profiles were studied. Results indicated that out of a total of 161 genes, 38 were differentially expressed by X-ray treatment and 24 by heavy ion (Fe(+2)) treatment. Eight genes were common to both treatments and were confirmed by Northern blot analysis: BRCA1, BIRC2/CIAP1, CENP-E, DDB1, MRE11A, RAD54/ATRX, Wip1 and XPF/ERCC4. A number of candidate genes reported here may be useful molecular biomarkers of radiation exposure in breast cells. PMID- 18425357 TI - Bufalin induces growth inhibition, cell cycle arrest and apoptosis in human endometrial and ovarian cancer cells. AB - Bufalin is a traditional Chinese medicine and it induces apoptosis in certain human tumor cell lines. We investigated the effect of bufalin on three endometrial cancer cell lines, two ovarian cancer cell lines, and on normal human endometrial epithelial cells. Endometrial and ovarian cancer cells were treated with various concentrations of bufalin, and its effect on cell growth, cell cycle, apoptosis, and related measurements was investigated. The 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay showed that all endometrial and ovarian cancer cell lines were sensitive to the growth-inhibitory effect of bufalin, although normal endometrial epithelial cells were viable after treatment with the same doses of bufalin that induced growth inhibition in endometrial and ovarian cancer cells. Cell cycle analysis indicated that their exposure to bufalin decreased the proportion of cells in the S-phase and increased the proportion in the G0/G1 phases of the cell cycle. Induction of apoptosis was confirmed by annexin V staining of externalized phosphatidylserine and loss of the transmembrane potential of mitochondria. This induction occurred in concert with the altered expression of genes related to cell cycle and apoptosis. These results suggest that bufalin may become a useful adjuvant therapy for endometrial and ovarian cancers with minimal side effects. PMID- 18425358 TI - p53-related apoptosis resistance and tumor suppression activity in UVB-induced premature senescent human skin fibroblasts. AB - Chronic exposure to solar UV irradiation leads to photoaging, immunosuppression, and ultimately carcinogenesis. Cellular senescence is thought to play an important role in tumor suppression and apoptosis resistance. However, the relationships among stress-induced premature senescence (SIPS), tumorigenesis and apoptosis induced by UVB remain unknown. We developed a model of UVB-induced premature senescence in human skin fibroblasts (HSFs). After five repeated subcytotoxic UVB exposures at a dose of 10 mJ/cm2, the following biomarkers of senescence were markedly present: senescence-associated beta-galactosidase (SA beta-gal) activity, growth arrest, and the overexpression of senescence associated genes. Firstly, there was an increase in the proportion of cells positive for SA beta-gal activity. Secondly, there was a loss of replicative potential as assessed by MTT assay. FACS analysis showed that UVB-stressed HSFs were blocked mostly in the G1 phase of the cell cycle, and replicative senescence, and protein expression of p53, p21(WAF-1) and p16(INK-4a) increased significantly. Thirdly, the mRNA levels of three senescence-associated genes, fibronectin, osteonectin and SM22, also increased. A real time PCR array to investigate the mRNA expression of p53-related genes involved in growth arrest, apoptosis and tumorigenesis indicated that p53, p21, p19, Hdm2, and Bax were up regulated, and bcl, HIF-1alpha and VEGF were down-regulated. Collectively, our data suggest that UVB-induced SIPS plays an important role in p53-related apoptosis resistance and tumor suppression activity. PMID- 18425359 TI - TP53 gene mutations as an independent marker for urinary bladder cancer progression. AB - This study evaluates the influence of the TP53 genetic status on tumour recurrence and progression with a highly effective electrophoretic technique. DNA from tissue of 75 non-invasive urinary bladder cancers was PCR amplified in the TP53 exons 5-8 and run on horizontal polyacrylamide gels under defined temperature conditions to yield specific gel shifts. Kaplan-Meier and Cox Regression analysis were performed with tumour progression. The overall tumour recurrence in our patient population was 76.0% (57/75). Tumour recurrence frequency was 69.4% (34/49) in patients with TP53 wild-type, and 88.5% (23/26) in patients with TP53 mutation. There was no statistically significant difference with regard to recurrence frequency and time to recurrence. The progression-free survival was significantly shorter in patients with TP53 mutations, and the frequency of tumour progression was significantly higher in mutated as compared to wild-type tumours. Cox-Regression analysis showed a significant and independent influence of TP53 mutation on tumour progression in comparison with tumour grade, stage and history of prior bladder cancer. If segregated by exons, mutations in the DNA binding region of exon 8 seem to have a particular high influence on tumour progression. We conclude that genetic analysis of TP53 can select patients at high risk of bladder tumour progression that should be followed closely and may benefit from early radical surgical procedures. PMID- 18425360 TI - Effect of combining anti-epidermal growth factor receptor antibody C225 and radiation on DU145 prostate cancer. AB - The epidermal growth factor receptor (EGFR) network has rich targets for prostate cancer killing. Herein we evaluated the effects of combining the EGFR inhibition and radiation on DU145 prostate cancer. We treated DU145 prostate cancer cells with various doses of anti-EGFR antibody (C225) and gamma-irradiation (RAD). The effects of the treatment on cell viability and growth were assessed with cell counting, XTT and clonogenic assays. In vivo treatment effects were assessed using a subcutaneous tumor xenograft in mice. Cell cycle distribution and progression were assessed with flow cytometry. The apoptotic components of cell death were quantified using Annexin-V binding assays. The results demonstrated that when combined with radiation, C225 augmented the inhibition of cell viability and growth in the DU145 cell line and EGFR inhibition appeared to have some interaction with RAD. C225 inhibited the growth of implanted DU145 tumors and increased the efficacy of radiation treatment. Flow cytometric analysis suggested that mostly necrotic cell death resulted from the EGFR inhibition or irradiation, although there may be some apoptosis. We drew the conclusion that the inhibition of EGFR augments the radiation killing of DU145 prostate cancer via a combination of cytostatic, necrotic and apoptotic mechanisms. PMID- 18425361 TI - Involvement of EphA2 in head and neck squamous cell carcinoma: mRNA expression, loss of heterozygosity and immunohistochemical studies. AB - EphA2 is a 130-kDa transmembrane protein primarily found in adult human epithelial cells and is a member of one of the largest receptor tyrosine kinases. It is located on 1p36.1, a genetic hot spot in cancer. EphA2 overexpression has been observed in aggressive solid tumors and its potential role in tumorigenesis, which includes cell growth, survival, migration and angiogenesis have been reported. However, the role of EphA2 remains unknown in head and neck cancer. In this study, we investigated the genetic profile of EphA2 in primary head and neck squamous cell carcinoma (HNSCC) by determining mRNA level, status of loss of heterozygosity and protein expression. mRNA expression was also correlated with clinicopathological data. Infrequent loss of heterozygosity (20%) was observed, though a 10-fold increase of mRNA expression in tumors compared to normal tissues was noted. A significant number of samples with normal to high mRNA expression was observed among patients with regional metastasis, with T3-T4 tumor size and with moderate to poor differentiation. However, statistical studies did not show any correlation between mRNA expression and any of the clinicopathological parameters. Tumor cells expressed EphA2 protein, but only weakly. These results suggest that EphA2 might be involved in the early development of HNSCC although not directly responsible for its progression. PMID- 18425362 TI - Secretory leukoprotease inhibitor inhibits cell growth through apoptotic pathway on ovarian cancer. AB - In light of the poor prognosis for ovarian cancer, research continues for innovative and efficacious treatment modalities. It is now widely accepted that new approaches for the treatment of ovarian cancers are pivotal in further improving prognosis of this disease. Secretory leukoprotease inhibitor (SLPI) is an 11.7-kDa non-glycosylated, serine protease inhibitor that has a broad inhibitory spectrum against serine protease. SLPI showed potential therapeutic inhibitory effects mediated by tumor necrosis factor (TNF)-related apoptosis inducing ligand (TRAIL), TNF-alpha, death receptor (DR)-4, DR-5 and TNF receptor (TNFR)-I expression which lead to an activation of apoptosis pathway through Caspase-2, Caspase-8 and Caspase-9. We examined whether levels of SLPI protein expression correlated with clinicopathological characteristics in 58 ovarian cancer samples, and investigated the role of SLPI and its biological functions. SLPI expression showed a significant correlation between low expression of SLPI and amount of ascites (p=0.021), lymph node metastasis (p=0.011). We propose that SLPI could be considered a therapeutic target for the treatment of ovarian cancer. PMID- 18425363 TI - Expression of E1AF, an ets-oncogene transcription factor, highly correlates with malignant phenotype of malignant melanoma through up-regulation of the membrane type-1 matrix metalloproteinase gene. AB - Matrix metalloproteinase (MMP) is closely involved in the degradation of extracellular matrix and confers invasive and metastatic potential to malignant tumors. MMP-2 is a type-IV collagenase secreted as a proenzyme that is activated on the surface of the tumor cell by membrane-type 1-MMP (MT1-MMP). MT1-MMP plays a critical role during tumor progression and metastasis. We investigated the expression levels of E1AF and MT1-MMP in malignant melanoma cell lines and specimens from patients in order to clarify the mechanisms responsible for the invasion and metastasis of malignant melanoma. High levels of E1AF and MT1-MMP mRNA expression were observed in melanoma cells by Northern blotting and real time PCR. The expression level was highly correlated with an invasive potential determined by an in vitro invasion assay. The down-regulation of MT1-MMP was identified when E1AF was knocked down by RNA interference. These results suggest that E1AF plays a crucial role in the invasion and metastasis of malignant melanoma through up-regulating the MT1-MMP expression. PMID- 18425364 TI - The activation of Akt during preoperative chemotherapy for esophageal cancer correlates with poor prognosis. AB - The activation of the PI3K/Akt/mTOR pathway plays an important role in tumorigenesis and resistance to anticancer drugs. The aim of this study was to elucidate the role of the Akt/mTOR pathway in chemoresistance and the prognosis of patients with esophageal squamous cell carcinoma (ESCC) who received preoperative chemotherapy. We evaluated p-Akt and p-mTOR expression by immunohistochemistry in the surgical specimens of 143 ESCC (51 patients with and 92 without preoperative chemotherapy). In 37 patients of the former group, paired tissue samples obtained before and after chemotherapy were examined immunohistochemically. The incidence of p-Akt expression was higher in ESCC with than without chemotherapy (51.0 vs. 25.0%, p=0.0018). Although p-Akt expression was not associated with an advanced tumor stage, a comparison between before and after chemotherapy demonstrated an increased p-Akt expression during chemotherapy (p=0.0348). The p-Akt expression did not correlate with survival in ESCC without chemotherapy, but was associated with poor prognosis in those with chemotherapy (p=0.0058). In particular, an increased p-Akt expression during chemotherapy was associated with poor survival (p=0.0022). Notably, the p-mTOR expression did not correlate with p-Akt expression (p=0.1482). The depth of the tumor invasion, clinical response and p-Akt expression correlated with the prognosis of 51 ESCC with chemotherapy. A multivariate analysis showed that p-Akt expression was the only independent predictor of poor prognosis in ESCC patients with chemotherapy. p-Akt expression increases after chemotherapy in ESCC and a high expression correlates with poor prognosis. Our results suggest that the activation of Akt is a potentially useful therapeutic target in ESCC patients treated with chemotherapy. PMID- 18425365 TI - Clodronate inhibits angiogenesis in vitro and in vivo. AB - The effects of amino-bisphosphonate clodronate on endothelial cell functions involved in angiogenesis, namely proliferation and morphogenesis on matrigel were tested in vitro, whereas its effects on angiogenesis were studied in vivo. This was performed by using the chick embryo chorioallantoic membrane (CAM) assay. In vitro, clodronate inhibited the endothelial cell proliferation in a dose dependent fashion, peaking at 30 microM. At the same concentration, clodronate inhibited the fibroblast growth factor-2 (FGF-2)-induced capillary-like tube formation in the morphogenesis assay on matrigel. In vivo, when tested with the CAM assay, clodronate again displayed the capability to inhibit FGF-2-induced angiogenesis. Overall, these results suggest that antiangiogenesis by clodronate can be used to treat a wide spectrum of angiogenesis-dependent diseases, including certain chronic inflammatory diseases and cancer. PMID- 18425366 TI - Formoterol fumarate and roxithromycin effects on muscle mass in an animal model of cancer cachexia. AB - Our study probed the effects of the beta-2 adrenergic agonist, formoterol and the macrolide antibiotic, roxithromycin, on muscle wasting in a well-characterized animal model of cancer cachexia. Female Wistar rats were inoculated with Yoshida AH130 ascites hepatoma (AH) cells to induce rapid and severe cachexia as demonstrated by wet weight determinations of the hearts, gastrocnemius muscles and carcasses. The control animals received saline (vehicle) inoculations. The AH inoculated rats were treated once daily for four days by i.p. injection with a vehicle control, 1 mg/kg formoterol, 5 and 50 mg/kg roxithromycin or 1 mg/kg formoterol plus 5, 25, 40 and 50 mg/kg roxithromycin. The saline-inoculated animals were treated by i.p. injection with vehicle control, 1 mg/kg formoterol, 5 and 40 mg/kg roxithromycin. As a result, formoterol alone reduced the loss of muscle mass in the AH-inoculated rats by approximately one-half, consistent with literature reports. Roxithromycin alone at 5 mg/kg did not affect muscle mass in the AH-inoculated rats. Roxithromycin given alone at 50 mg/kg reduced the loss of muscle mass in AH-inoculated animals by approximately one-half. With respect to the antagonizing muscle loss, formoterol combined with either 5 or 25 mg/kg roxithromycin did not reach statistical significance versus formoterol alone, while formoterol plus either 40 or 50 mg/kg roxithromycin enhanced protection against muscle loss versus formoterol alone. The gastrocnemius weights in the AH inoculated rats treated with formoterol combined with 40 mg/kg roxithromycin were not significantly different from the muscle weights in the saline-inoculated controls. To sum up, formoterol and roxithromycin apparently exert anti-cachectic effects in an additive fashion and may offer the potential for combination therapy in cachexia. PMID- 18425367 TI - Irinotecan HCl, an anticancer topoisomerase I inhibitor, frequently induces ovarian failure in premenopausal and perimenopausal women. AB - The effects of irinotecan HCl (CPT-11) combination chemotherapies on the hypothalamus-pituitary-ovary endocrine system were examined clinically. The incidences of typical menopausal malaises and/or endocrinological findings were investigated in 32 gynecological cancer patients treated by CPT-11 combination chemotherapies. Patients who complained of menopausal malaises or had been treated by hormone replacement therapy before chemotherapy were excluded from the study. Menopausal malaise-like symptoms (MMLS) appeared in 6 of 32 patients (18.8%) during CPT-11 combination chemotherapy, and these symptoms were completely cured within a few days by administration of conjugated estrogen tablets (0.625 mg/day). All the MMLS cases were perimenopausal patients (47-57 years of age), and MMLS were not found in any of the postmenopausal patients who had exceeded 3 years since endocrinological menopause or patients who had recurrent cancer after pelvic radiotherapy. After exclusion of these 3-year postmenopausal patients and postirradiation patients, 6 of 7 patients aged 45-59 years complained of MMLS during CPT-11 combination chemotherapy. The incidence of CPT-11-induced MMLS showed no relationships with the anticancer drugs combined with CPT-11, mean total CPT-11 dose, mean number of CPT-11 injections, mean individual CPT-11 dose, grade of CPT-11-specific diarrhea or anticancer effects of each CPT-11 combination chemotherapy. The perimenopausal cancer patients with CPT-11-induced MMLS showed decreased serum estradiol and increased serum FSH and LH levels accompanying the CPT-11 injections. A young patient with CPT-11-induced secondary amenorrhea showed decreased serum estradiol and increased serum FSH and LH levels accompanying the CPT-11 injections. None of the postmenopausal patients with high FSH and LH levels showed any significant differences in their serum FSH, LH, PRL and TSH levels during CPT-11 combination chemotherapy. No differences in the results of LHRH and TRH tests during chemotherapy were found for postmenopausal patients. Histopathological examinations of normal ovarian tissues surgically removed from 4 young cervical cancer patients treated with preoperative CPT-11 combination chemotherapies revealed no growing ovarian follicles in the ovarian tissues. CPT-11 injections can induce estrogen-rescued MMLS in cancer patients aged approximately 50 years at a very high rate and may induce secondary amenorrhea in young women. The endocrinological and histopathological studies revealed that CPT-11 causes ovarian follicular loss and ovarian failure within a short time without affecting hypothalamic and pituitary hormone secretion. These clinical results indicate that CPT-11 has strong ovarian toxicity and that repeated CPT-11 administrations may frequently induce ovarian follicular loss and premature ovarian failure, even in young women. PMID- 18425368 TI - Improved detection of hepatic metastases of adrenocortical cancer by contrast enhanced ultrasound. AB - Adrenocortical carcinoma (ACC) is a rare and heterogeneous malignancy whose pathogenesis and poor prognosis is poorly understood. Computerized tomography (CT) and magnetic resonance imaging (MRI) are routinely performed for the imaging of the adrenal mass and for standard staging of the chest and abdomen as the lung and liver are the primary organs for metastasis in ACC. Contrast-enhanced ultrasound has been shown to have a high sensitivity and specifity for the differentiation of hepatic and neuroendocrine tumors. Twelve patients (7 women and 5 men; aged 24-74 years) with ACC were treated in our centre from 2004 to 2006. The patients received staging with a contrast-enhanced multislice spiral computed tomography (MSCT) as well as with a conventional and an echo-enhanced ultrasound of the liver. Contrast-enhanced ultrasound demonstrated liver metastases in 8 out of 12 patients (67%) and MSCT in 6 out of 12 patients (50%). In 2 out of 8 patients (25%) MSCT did not detect the liver metastases. Even in retrospective analysis with knowledge of the ultrasound results, the hepatic lesions were not recognized by the MSCT, but became detectable by MSCT at a later time point. All hepatic lesions diagnosed by MSCT were also seen by ultrasound. The detection of liver metastases by ultra-sound resulted in a change of therapy in two patients. The contrast-enhanced ultrasound has a high sensitivity in detecting the highly-vascularized liver metastases of ACC and should be included in the staging algorithm of ACC. PMID- 18425369 TI - PTEN and p27Kip1 are not downregulated in the majority of renal cell carcinomas- implications for Akt activation. AB - Activation of the PKB/Akt pathway is supposed to substantially contribute to the pathogenesis and progression of malignant disease. The present study aimed at determining the occurrence of an impaired PTEN and p27Kip1 expression alone or in combination in a renal cell carcinoma to further clarify the role of Akt-pathway associated proteins for the development and/or progression of this malignant disease. By using tissue microarray analysis, tissue samples from renal cell cancers and the corresponding benign tissue samples were investigated for expression of the PTEN, pAkt and p27Kip1 protein by immunohistochemistry. Additionally, a Western blot and RT-PCR analysis was performed to verify the results obtained from the immunohistochemical approach and to further clarify the mechanisms underlying the regulation of both proteins in renal cell cancer. Western blot analysis revealed an overexpression of PTEN and p27Kip1 in renal cell cancer samples and a significantly elevated expression of both proteins when compared with the corresponding benign tissue (p<0.0001 and p<0.0005). The latter finding was confirmed by real-time RT-PCR (p<0.05 and p<0.01) and immunohistochemistry (p<0.001 and p<0.0001). PTEN and p27Kip1 expression were positively correlated with each other both in the tumour and benign tissue (p<0.001 and p<0.0001). We concluded that a strong expression of PTEN in renal cell cancer did not block the PI3K-mediated phosphorylation of Akt in the tumour specimens analysed. Furthermore, Akt activation may not result in a decreased p27Kip1, the latter being retained and overexpressed in the majority of renal cell cancers when compared with the corresponding benign renal parenchyma. PMID- 18425370 TI - Expression and promoter methylation of the RASSF1A gene in sporadic breast cancers in Chinese women. AB - The novel tumor suppressor RASSF1A is frequently inactivated during human tumorigenesis by promoter methylation. In this study, we detected the RASSF1A promoter methylation by methylated-specific PCR and investigated RASSF1A gene expression by semi-quantitative RT-PCR and immunohistochemical staining in 36 cases of breast cancer and their adjacent normal tissues in Chinese women. The promoter methylation of the RASSF1A gene was found to be a frequent event in the breast cancers (61.1%). RASSF1A methylation was not found in the matched adjacent normal tissues. The loss frequency of RASSF1A mRNA was 33.3% and that of the RASSF1A protein was 44.4% in breast cancers. RASSF1A mRNA and protein were all expressed in adjacent normal tissues. The mRNA and protein expression level of RASSF1A was significantly lower in breast cancer than in adjacent normal tissue. However, the promoter methylation of the RASSF1A gene in breast cancers were not correlated with clinical parameters, such as ages, histological types, TNM stages and lymph node metastases. Thus, the promoter methylation of RASSF1A was one reason for the low level of RASSF1A mRNA and protein expression and was a frequent event in primary sporadic breast tumorigenesis in Chinese women. PMID- 18425371 TI - 1p31, 7q21 and 18q21 chromosomal aberrations and candidate genes in acquired vinblastine resistance of human cervical carcinoma KB cells. AB - Vinblastine (VBL) is used to treat certain kinds of cancer including Hodgkin's lymphoma, lung cancer, breast cancer, testicular cancer and cervical carcinoma. However, the rapid development of resistance during therapy remains a major clinical challenge. In order to reverse cancer cell resistance, the goal of this study was to find differentially expressed genes and chromosomal alterations in multidrug resistant (MDR) KB-v1 cells, further to probe the relationship between drug resistance and differential genes, and chromosomal changes in MDR cancer cells. Comparative genomic hybridization (CGH) analysis of MDR KB-v1 and their parental KB-3-1 cells revealed chromosomal changes; microarray-based expression profiling was carried out by comparing the gene expressions of MDR KB-v1 cells and KB-3-1 cells. We have identified 3 chromosomal gains in regions of 1p31, 7q21 and 18q21 in MDR cells and 10 genes (CYR61, UGTREL7, MBD1, NARS, ATP5A1, ABCB1, ABCB4, PEG10, MCM7, SERPINE1) contained in these regions were also up-regulated in MDR KB-v1 cells. Forty-nine genes were down-regulated when KB-v1 cells were subjected to lower dose or depletion of the drug. We have confirmed some gene expression changes by reverse transcription-polymerase chain reaction and Northern blots. These are the first data describing the relationship of 1p31 and 18q21 chromosomal aberrations and candidate genes in acquired vinblastine resistance. This study also demonstrates that the combination of CGH and cDNA microarray is a very useful tool to detect drug resistant targets in cancer treatment. PMID- 18425372 TI - Sodium salicylate switches glucose depletion-induced necrosis to autophagy and inhibits high mobility group box protein 1 release in A549 lung adenocarcinoma cells. AB - Sodium salicylate, the active metabolite of aspirin, has been shown to exert anti inflammatory activities by inhibiting the expression of various pro-inflammatory factors, and has potent anti-cancer effects against a number of human cancers including colon, lung, breast and leukemia. Necrotic cell death is emerging as one of the crucial factors that trigger an inflammatory response since during necrotic death the cell membrane is ruptured and the intracellular constituents including high mobility group box 1 (HMGB1) are released into the extracellular space, thereby activating an inflammatory response. In contrast, autophagic death is regarded as a form of tumour suppressive cell death, as indicated in tumour suppressors such as beclin 1 in autophagic pathways. To better understand the anti-inflammatory properties of sodium salicylate and its effect on necrotic cell death in A549 cells induced by glucose depletion (GD), a common characteristic of the tumour micro-environment, was examined. While GD induced mostly necrotic death in A549 cells, salicylate suppresssed GD-induced necrosis and HMGB1 release. In addition, salicylate shifted the cell death pattern to autophagy by inhibiting GD-induced Cu/Zn superoxide dismutase release and ROS production. These results indicate that the activity of salicylate to prevent necrotic death may contribute to its anti-inflammatory action and suppress tumour development possibly through switching the cell death mode from tumour-promoting necrotic cell death to tumour-suppressive autophagic cell death. PMID- 18425373 TI - Interleukin 15 augments antitumor activity of cytokine gene-modified melanoma cell vaccines in a murine model. AB - Many studies have demonstrated that interleukin 15 (IL-15) is a cytokine with strong antitumor properties and have suggested its potential use in tumor immunotherapy. IL-15 exerts its effect on innate and acquired immunity with the most prominent action in NK cells and CD8(+) memory T cells. Therefore, many authors have proposed that IL-15 could be a good candidate for augmenting the efficacy of vaccination strategies. In our experiments, in a model of B78-H1 murine transplantable melanoma, tumor-bearing mice were treated with different cytokine-gene modified tumor cell vaccines (producing TNF-alpha, GM-CSF, IL-12 or IL-6/sIL-6R) followed by a series of IL-15 injections. In order to investigate the infiltration of treated tumors by leukocytes, immunohistochemical staining was performed. In every case, the combined therapy was superior to the treatment with either a vaccine or IL-15 alone. Tumors treated with the combination of B78 H1 melanoma cells secreting IL-12 (B78/IL-12 vaccine) and IL-15 were heavily infiltrated by granulocytes. IL-15, either alone or in combination with the B78/IL-12 vaccine, influenced infiltration of tumors with CD3(+) lymphocytes, CD4(+)and CD8(+). To our knowledge, this is the first report that shows the universal genetically-modified tumor cell vaccine-augmenting properties of IL-15. The cytokine can be useful as an adjuvant in cancer gene therapy in humans. PMID- 18425374 TI - A dual colour dual fusion fluorescence in situ hybridisation study on the genesis of complex variant translocations in chronic myelogenous leukaemia. AB - Complex variant 9;22 translocations occur in a significant minority of chronic myelogenous leukaemia (CML) patients. Different mechanisms of their formation have been described. We report dual colour dual fusion fluorescence in situ hybridisation data in 12 Chinese CML patients with complex translocations. Three previously reported breakpoint hotspots in a third partner chromosome (14q32, 17q25, 1q21) were observed. In 10/12 (83.3%) patients, the abnormality occurred as a single step 3-break event. Only a single abnormal clone harbouring the complex translocation was seen in this group. The remaining 2 cases in the chronic phase showed a 4-break mechanism (2/12, 16.7%). Deletion of 5' ABL at der(9) was not observed in any of the 12 patients, however, the loss of 3' BCR was observed in 1 patient (1/12, 8.3%). Together with previous findings, these data suggest that these variant translocations occur more often as a 3-break single-step process with no reciprocal ABL-BCR fusion. On the other hand, a 4 break event is also regularly seen during the initial stages of leukaemogenesis, which likely predisposes to der(9) deletion. The observed difference in rates of der(9) deletion reported in a series of CML patients with variant translocations may be related to a difference in rates of a 4-break event. PMID- 18425375 TI - Expression of Sonic hedgehog signaling pathway correlates with the tumorigenesis of intraductal papillary mucinous neoplasm of the pancreas. AB - Sonic hedgehog (SHH) is frequently expressed in pre-cancerous lesions and carcinoma of the pancreas. A recent study revealed that its expression was higher in the intraductal papillary mucinous neoplasm (IPMN) of the pancreas than in the pancreatic carcinoma. However, the correlation between its signaling pathway and tumorigenesis of IPMN has not yet been well documented. We investigated the expression of mRNA and protein of SHH as well as its downstream transcription factor Gli1 in 19 microdissected lesions from 15 cases and in 75 lesions from 33 cases of the IPMN by one-step quantitative real-time reverse transcription polymerase chain reaction and immunohistochemistry, respectively. SHH and Gli1 mRNAs were detected in all the examined lesions and 8 out of 19 lesions in IPMNs, respectively. SHH and Gli1 mRNAs were likely to be up-regulated from the adenoma and from borderline to carcinoma cells, respectively. Immunohistochemical analysis also reported that SHH and Gli1 expression was correlated with the grade of cell atypia. These findings suggested that HH signaling was activated in IPMNs and contributed to tumorigenesis in these types of neoplasms. PMID- 18425376 TI - New insight into the key proteins and pathways involved in the metastasis of colorectal carcinoma. AB - Metastasis-associated genomic alterations have been recognized to play a critical role in tumor metastasis. Primary and metastatic tumor cells in mice and tumors in a patient were studied by cDNA array analysis. Selected genes were determined by RT-PCR and immunohistochemistry. Pathways on changed genes were statistically analyzed. The function of Grb2 was determined by in vitro wound assay. Nodal metastatic cells had a stronger ability of growth and metastasis than primary tumor cells. A total of 376 genes showed a different expression between primary and metastatic cells. The expression of Grb2 and genes in the Grb2-mediated pathways was significantly elevated in the metastases. Elevated levels of Grb2 expression in metastases were related to the distant metastasis of colorectal carcinoma. Blocking the Grb2-SH2 domain signaling transduction inhibited cell motility. Metastasis-associated genes identified by cDNA and tissue microarrays provide potentially valuable information on the metastasis of colorectal tumors. Overexpression of Grb2 may contribute to tumor growth, invasiveness and metastasis. PMID- 18425377 TI - Lovastatin enhances the antitumoral and apoptotic activity of doxorubicin in murine tumor models. AB - Despite its effectiveness as an antineoplastic drug, doxorubicin (DOX) is usually associated with cardiotoxicity. Lovastatin (LOV), a hypolipidemic agent used in the clinic, has been demonstrated to have antitumoral and antimetastatic effects in murine models. Since the two agents arrest tumor cells in different phases of the cell cycle and induce apoptosis, the goal of this study was to examine the efficacy of a combination therapy with LOV and low doses of DOX, in an attempt to obtain an improved antitumoral effect devoid of toxicity, by using a rat B-cell lymphoma and a mouse mammary tumor. In the two models, the combined treatment showed a synergistic antitumoral effect, which is mainly ascribed to an increased apoptotic response elicited by a LOV/DOX combination than either agent alone. The therapeutic benefit demonstrated by the combination treatment is further emphasized by the lack of toxicity. PMID- 18425378 TI - First genetic analysis in Tunisian familial adenomatous polyposis probands. AB - Familial adenomatous polyposis (FAP) is an autosomal dominant inherited disease characterized by the development of hundreds to thousands of adenomatous polyps in colon and rectum. The APC gene (adenomatous polyposis coli) is considered as the major mutated gene in FAP. It has been shown that biallelic germline mutations in the base-excision-repair gene MYH can be responsible for a recessive inheritance of adenomatous polyposis (AP). This study is the first Tunisian genetic analysis on AP patients. Multiplex ligation-dependent probe amplification (MLPA) was used to screen the APC gene for large genomic rearrangements. The total APC and MYH exon sequences and exon-intron edges were sequenced in an effort to detect germline mutations, four were explored. Mutations were detected in four patients that fulfil the clinical criteria of AP. Three mutations were found in the APC gene, of which two were novel (c.1636_1639delAGTG and c.2514 G>T) and all gave rise to a truncated APC protein. The missense G382D mutation, already described in north and south European populations was found in the MYH gene at the homozygous state in the fourth patient with moderate AP. Our preliminary study provides a basis for implementation of genetic counselling for AP. PMID- 18425379 TI - Intracavitary chemotherapy (Gliadel) and oral low-dose etoposide for recurrent anaplastic ependymoma. AB - Anaplastic ependymoma is associated with a higher incidence of tumor recurrence and its prognosis still remains unsatisfactory. Consolidated therapy for ependymoma includes surgery followed by focal radiotherapy when resection is incomplete. In the case of relapse treatment, options are limited especially for patients who have already received radiotherapy. We sought to establish the feasibility of administering low-dose oral etoposide (50 mg/m(2)/day for 21 days) in combination with the implantation of intracavitary carmustine (BCNU) wafers (Gliadel) at the gross total resection for achieving synergistic treatment in three children affected by recurrent anaplastic ependymoma. All patients had Karnofsky performance scale (KPS) scores >80%. The therapy was tolerated safely and well in all patients without any post-surgery complications. After BCNU wafer implantation, all patients achieved radiological and clinical stabilization for an average period of 3 months. Two patients relapsed after 4 months as shown in brain MRIs. The other patient went to progression two months after the Gliadel implantation. This multimodal approach was not effective for the treatment of refractory anaplastic ependymoma and further studies are required in order to define the role of the combination of multidrug systemic chemotherapy with BCNU wafer implantation in children with high-risk brain tumors. PMID- 18425380 TI - Inhibition of NADPH oxidase activity promotes differentiation of B16 melanoma cells. AB - The activity of NADPH oxidase is increased in malignant skin keratinocytes. We demonstrated that inhibition of NADPH oxidase activity by diphenyleneiodonium (DPI) suppressed free radical production, inhibited cell growth and promoted cell differentiation of B16 melanoma cells, as indicated by cell morphology, increased production of melanin, and increased expression of microphthalmia-associated transcription factor (MITF). siRNA to NADPH oxidase subunit Rac1 or p47 induced the expression of MITF, verifying that the pro-differentiation effects are due to the inhibition of NADPH oxidase. Biochemical studies suggest that ERK plays a positive role whereas PKCalpha plays a negative role during this differentiation event. In addition, the protein levels of the tumor suppressor p53 were suppressed by DPI, suggesting that p53 is activated by oxidative stress and may negatively regulate differentiation in melanoma cells. Taken together, these results suggest that inhibiting NADPH oxidase activity promotes cell differentiation of B16 melanoma cells. PMID- 18425381 TI - Genistein differentially modulates androgen-responsive gene expression and activates JNK in LNCaP cells. AB - Genistein, the predominant isoflavone in soy, may be chemopreventive in prostate cancer (CaP). It down-regulates the prostate-specific antigen (PSA) and androgen receptor (AR) in androgen responsive cells. However, the extent of the down regulation and whether genistein has a general effect on all androgen responsive genes (ARGs) are unclear. We investigated the ability of genistein to modulate ARG expression by the synthetic androgen R1881 in LNCaP cells. Given that there is important crosstalk between AR and mitogen activated protein kinase (MAPK) signaling, we also investigated whether genistein activates the MAPK end targets c-Jun N-terminal kinase (JNK) and c-Jun. Changes in ARG expression were determined by Western analysis and semi-quantitative RT-PCR. The activation of JNK and c-Jun was investigated by Western analysis and a solid phase kinase assay. The PSA protein and mRNA expression were both down-regulated by genistein. In contrast, KLK4 was up-regulated at the mRNA, but down-regulated at the protein level. NKX3.1 mRNA levels did not change significantly, but protein levels were significantly down-regulated. STAMP2 mRNA levels slightly increased whereas the protein expression was down-regulated. The AR mRNA expression changed significantly only at high concentrations of genistein when it was down regulated, whereas AR protein levels were decreased at low concentrations of genistein. The solid phase kinase assay indicated a transient activation of JNK by genistein, which was supported by Western analysis. Thus genistein differentially modulates ARG mRNA expression, but has an inhibitory role on the ARG protein levels. The activation of the JNK pathway which inhibits AR signaling may provide a mechanism for the overall inhibition of protein levels. PMID- 18425382 TI - 2-DE analysis of breast cancer cell lines 1833 and 4175 with distinct metastatic organ-specific potentials: comparison with parental cell line MDA-MB-231. AB - Human MDA-MB-231 derived breast cancer cell lines 1833 and 4175 have different metastatic potentials in terms of their tissue tropisms and aggressiveness. Cell line 1833 is specifically metastatic to the bone. The highly aggressive cell line 4175 is specific to the lung. We performed 2-DE analysis of the cell lines. We found 16 significantly changed protein spots, 14 protein spots were identified. Expression of cathepsin D, triosephosphate isomerase, phosphoglycerate kinase 1, heme binding protein 1 and annexin 2 could be correlated with the in vitro aggressiveness of the respective cell lines. Interstitial collagenase and dimethylargininase 2 were exclusive to the cell line 1833 and might contribute to its bone specificity. Serpin B9, cathepsin B chain b, galectin 3 and HSP 27 were changed in the lung specific cell line 4175. The possible contribution of identified proteins to differences in metastatic behavior of the cell lines is discussed. PMID- 18425383 TI - Role of serum-derived hyaluronan-associated protein-hyaluronan complex in ovarian cancer. AB - The objective of this study was to determine if the level of serum hyaluronan (HA), serum-derived HA-associated protein (SHAP)-HA complex, and urinary trypsin inhibitor (UTI) correlate with the clinical outcome of ovarian cancer patients. The relationship of metalloproteinase and its inhibitor with HA and the SHAP-HA complex was also examined. Serum and urine samples were obtained from 45 patients with ovarian cancer, 22 patients with benign ovarian tumors and 50 healthy women. Concentrations of serum HA and UTI were measured by an inhibitory sandwich enzyme linked immunosorbent assay, and concentrations of the serum SHAP-HA complex were measured by a sandwich enzyme-linked immunosorbent assay. Concentrations of MMP 2, MMP-9 and TIMP-1 were measured by a one-step enzyme immunoassay. The levels of HA, SHAP-HA complex, MMP-9 and TIMP-1 were higher in the ovarian cancer group than in the benign ovarian tumor group. In ovarian cancer patients, the levels of HA, SHAP-HA complex and MMP-9 were higher in the stage III/IV group than in the stage I/II group, and the levels of SHAP-HA complex, MMP-9 and TIMP-1 were higher in the non-responder group than in the responder group. The serum concentration of SHAP-HA complex had a significant correlation with HA, MMP-9 and TIMP-1 in ovarian cancer patients. The patients with elevated SHAP-HA complex had a shorter disease-free survival compared with those with normal levels of SHAP-HA complex. The multiple regression analysis revealed that SHAP-HA complex is the significant independent variable for progression-free survival. The elevated level of SHAP-HA complex may indicate the prognosis of recurrence and reflect the tumor metastasis associated with MMP-9 in ovarian cancer patients. PMID- 18425384 TI - Suppression of the proliferation of cancer cell lines, KB-3-1 and K562 cells preceded by a decrease in intracellular pH caused by phenoxazine derivatives. AB - The intracellular pH (pHi) of cancer cells such as the KB-3-1 (human epidermoid carcinoma cell line) and K562 cells (human chronic myeloid leukemia cell line) cultured in a medium (pH 7.4) was found to be much higher (pH 7.65 and 7.8, respectively) than that of normal cells (pHi is usually < or =7.2). When a phenoxazine derivative, 2-aminophenoxazine-3-one (Phx-3) or 2-amino-4,4alpha dihydro-4alpha-7-dimethyl-3H-phenoxazine-3-one (Phx-1) was added to these cells, pHi rapidly decreased within 20 min, dose-dependently, though the extent of the decrease of pHi was significantly larger for Phx-3 (a decrease of 0.9 units) than for Phx-1 (a decrease of 0.4 units). Phx-3 and Phx-1 caused the proliferative suppression of these cells 24 h after the addition, dose-dependently. The anti proliferative effects of Phx-3 on KB-3-1 and K562 cells were far greater than those of Phx-1. It was proposed that the proliferative suppression of KB-3-1 and K562 cells caused by Phx-3 and Phx-1, may be preceded by a rapid and extensive decrease in pHi, which possibly influenced the intracellular homeostasis, finally causing the suppressed proliferation and apoptosis of these cancer cells. The present results suggest that the anti-cancer effects of Phx-3 and Phx-1 may be strengthened by the intracellular acidification of cancer cells by these compounds. PMID- 18425385 TI - Survival analysis of ovarian clear cell carcinoma confined to the ovary with or without comprehensive surgical staging. AB - Pure-type clear cell carcinoma (CCC) has been recognized as a distinct subtype of ovarian cancer, showing a resistance to chemotherapy and resulting in poor prognosis. Our aim was to evaluate the effects of complete surgical procedures followed by adjuvant chemotherapy for CCC patients whose tumors were confined to the ovary (pT1M0). During the period of 1987-2005, 56 patients with stage I CCC were identified and two cases were excluded due to retroperitoneal lymph node metastasis. A total of 54 patients were enrolled in the study and divided into two groups: Group A (n=38, 1993-2005) underwent complete surgical staging including pelvic and para-aortic lymphadenectomy. Group B (n=16, 1987-1992) underwent a hysterectomy, bilateral salpingo-oopherectomy, omentectomy without comprehensive lymphadenectomy. Every patient received six courses of adjuvant chemotherapy using a platinum agent. Survival analysis was estimated by the Kaplan-Meier method and prognostic factors were evaluated using a Cox regression model. The clinical characteristics of the two groups were similar, except for the rate of conventional platinum-based chemotherapy (p=0.02). Multiple regression survival analysis revealed that the completion of a comprehensive staging operation was the only independent factor for progression-free survival of stage I CCC patients (p=0.03) and that the chemotherapeutic regimen was not a prognostic factor (p=0.43). The present study indicates that we should accomplish complete surgical staging procedures for CCC confined to the ovary. PMID- 18425386 TI - Combinations of tumor-specific CD8+ CTLs and anti-CD25 mAb provide improved immunotherapy. AB - One new approach to cancer therapy is based on the adoptive transfer of tumor specific cytotoxic T cells and anti-CD25 antibodies. In the present study, CD8+ and IFN-gamma secreting T lymphocytes (CTLs) were enriched as tumor-specific cytotoxic T cells from spleen lymphocytes of mice bearing the Renca tumor (a murine renal carcinoma line originating from a BALB/c mouse) after stimulation with tumor cells. An anti-CD25 IL-2Ralpha(anti-CD25) mAb from hybridoma PC61 was used for depletion for CD4(+)CD25(+) regulatory T (Treg) cells. Treatment efficacy for tumor-bearing mice was compared using 4 systems: 1, whole spleen lymphocytes stimulated with tumor cells in vitro from tumor-bearing mice; 2, CTLs; 3, anti-CD25 mAbs; 4, CTLs and anti-CD25 mAbs. At the 50th day after tumor inoculation, in the group which received anti-CD25 mAb for depletion of T cells and inoculation of CTLs, tumors had disappeared and no re-growth was observed. In contrast, all mice of the non-treated and other three groups, treated with whole spleen cells alone, CTLs alone and anti-CD25 mAb alone, had died. These results showed that a combination of Treg cell-depletion using anti-CD25 mAbs and CTL administration is a feasible approach for treatment of cancers which warrants further exploration in the clinical setting. PMID- 18425388 TI - Identification of EphB6 variant-derived epitope peptides recognized by cytotoxic T-lymphocytes from HLA-A24+ malignant glioma patients. AB - We found previously that EphB6, a member of the erythropoietin-producing hepatocyte (Eph) receptor tyrosine kinase family, was preferentially expressed in malignant gliomas. In the present study, RT-PCR revealed a putative secretory variant form of human EphB6 that was expressed in the majority of glioma cell lines, though not in normal tissues. The variant has a unique 54 amino acid sequence that is not found in the normal EphB6. Therefore, we attempted to determine the antigenic peptides unique to the variant for immunotherapy. The two variant-derived peptides had the ability to bind to HLA-A2402 molecules and each of them could induce cytotoxic T-lymphocytes (CTLs) in vitro in peripheral blood mononuclear cells of HLA-A24(+) glioma patients. Furthermore, the cytotoxicity was mediated by peptide-specific CD8(+) T cells in an HLA-A24 restricted manner. Taken together, the two peptides derived from the variant of EphB6 might be appropriate targets for peptide-based specific immunotherapy to HLA-A24(+) patients with malignant glioma. PMID- 18425387 TI - Fluorescent in situ hybridization as a screening test for HER2 amplification in G2 and G3 breast cancers of lobular and ductal histotype and metastases. AB - The aim of the present study was to evaluate the effectiveness of fluorescence in situ hybridisation (FISH), as a screening test, in moderately- (G2) or poorly- (G3) differentiated breast cancers of the ductal (IDC) and lobular (ILC) histotypes and distant metastases. HER2 FISH was performed on 486 G2 and 477 G3 both of IDC and ILC histotypes and in 241 metastases. A significant difference in the HER2 amplification was observed between G2 (14.8%) and G3 (31.9%), with no difference according to the histotype. However, the rate of amplification increased to 36% in the G2/hormone receptor-negative cases as compared to 10.6% in the G2/receptor-positive cases (p<0.0001). HER2 was amplified in 17% of metastases with some differences depending on the location. These data suggest that the HER2 FISH analysis may be an effective screening test in breast cancer metastases and G3 tumors, irrespective of the hormone receptor status or presence of lymphovascular invasion. PMID- 18425389 TI - Clinicopathological significance of the gene expression of matrix metalloproteinases and reversion-inducing cysteine-rich protein with Kazal motifs in patients with colorectal cancer: MMP-2 gene expression is a useful predictor of liver metastasis from colorectal cancer. AB - Matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9 (MMP-9) and membrane-type matrix metalloproteinase 1 (MT1-MMP) are involved in colorectal cancer invasion and metastasis. Reversion-inducing cysteine-rich protein with Kazal motifs (RECK) inhibits MMP-2, MMP-9 and MT1-MMP. We examined the clinicopathological significance of the relative expression of these genes in patients with colorectal cancer, especially with regard to metastasis. We studied surgical specimens of cancer tissue and adjacent normal mucosa obtained from 205 patients with untreated colorectal carcinoma. MMP-2, MMP-9, MT1-MMP, RECK and beta-actin mRNA of cancer tissue and adjacent normal mucosa were measured by quantitative real-time reverse-transcriptase polymerase chain reaction. MT1-MMP gene expression was higher in cancer tissue than in adjacent normal mucosa. In contrast, MMP-2, MMP-9 and RECK gene expression levels were lower in cancer tissue than in adjacent normal mucosa. As for the relationship between the gene expression and clinicopathological factors, MMP-2 expression correlated with the depth of invasion, venous invasion and liver metastasis; MMP-9 and RECK expression correlated with venous invasion. There were positive correlations among the gene expression levels of MMP-2, MMP-9 and RECK. MMP-2 gene expression was considered a useful predictor of liver metastasis from colorectal cancer. PMID- 18425390 TI - Alterations of the K-ras and p53 genes in Tamoxifen-associated endometrial carcinoma. AB - To better understand the molecular mechanisms of carcinogenesis induced in uterine endometrium by therapeutic anti-estrogenic Tamoxifen (TAM) exposure, 27 uterine tumors (4 benign endometrial polyps and 23 carcinomas) associated with TAM exposure were analyzed for the presence and spectrum of p53 and K-ras mutations. Although there was no significant difference between TAM-associated endometrial carcinomas and sporadic endometrial tumors in the frequency of these mutations, the spectrum of p53 mutations was characteristically unique to the TAM associated tumors. The median duration of TAM exposure was significantly longer in patients with p53 mutations than those without p53 mutations (62 vs. 30 months, p=0.028). Our observation suggests that prolonged TAM exposure may directly inactivate the p53 gene by acting as a mutagen in a significant fraction of TAM-associated endometrial carcinomas. PMID- 18425391 TI - Perforating branches of the internal thoracic artery in women with breast cancer: an anatomical study for breast-conservation surgery. AB - Breast reconstruction after breast-conservation surgery is an important issue for breast cancer patients. Various factors are associated with complications, of which blood flow is one of the more important. The perforating branches of the internal thoracic artery (ITA) are key contributors to blood flow in the anterior chest wall. The present study examined the distributions and depths of the perforating branches of the internal thoracic artery using a multi-detector row computed tomography (MDCT) angiography. The subjects in this prospective study comprised of 38 women with suspected breast cancer who underwent MDCT angiography. The images were analyzed on computer using transverse MDCT source data and volume renderings. A total of 47 perforating branches were found, with 27 (57.4%) originating in the second, 6 (12.8%) in the third and fourth, 5 (10.6%) in the first and 2 branches (4.3%) originating in the fifth intercostal space, with one branch (2.1%) originating opposite the first intercostal space. A strong correlation was identified between the distance from the skin to the branch and adipose thickness at the shallowest and deepest points (P<0.001). The distributions and depths of the perforating branches of ITA identified in this study may be helpful in immediate breast reconstruction following mastectomy or breast-conserving surgery. PMID- 18425392 TI - Dual anti-cancer effects of weekly intraperitoneal docetaxel in treatment of advanced gastric cancer patients with peritoneal carcinomatosis: a feasibility and pharmacokinetic study. AB - This study evaluated the feasibility and pharmacology of intraperitoneal docetaxel (IP docetaxel) when administered weekly for 3 consecutive weeks, followed by 1 week without treatment. A total of 24 patients with peritoneal carcinomatosis of gastric cancer (10 preoperative, 7 postoperative and 7 recurrent) were enrolled in this study. Docetaxel was dissolved in an isotonic saline to a final 1 liter solution and was administered in a 1 h dosage of 25, 35, 45 and 60 mg/m(2) to determine the maximum tolerated dose (MTD). To measure the docetaxel concentration, blood and peritoneal fluid samples were collected 0.5, 1, 2, 3, 6 and 24 h after administering the drug to 15 patients. A total of 109 chemotherapy cycles were administered, with a median of four cycles per patient (range 2-9). The MTD of the weekly IP docetaxel was defined at 60 mg/m(2). At a docetaxel dosage of 60 mg/m(2) per week, the dose-limiting events of grade 3 abdominal pain and grade 3 diarrhea, which may be associated with local toxicity, occurred. Peak concentrations of peritoneal fluid ranged from 24.5 to 68.7 microg/ml. The mean ratio of the area under concentration (AUC) in the peritoneal fluid to the plasma concentration was 515. Furthermore, the mean of plasma AUC by IP docetaxel was 5.63 microg h/ml versus that of IV docetaxel at a dose of 60 mg/m(2). The response rate of the preoperative IP docetaxel was 80% (4 CR, 4 PR, 1 NC and 1 PD), which was judged with laparoscopy and peritoneal lavage cytology. Gastrectomy, with D2 lymph node dissection, was performed on all of the patients evaluated as CR. The weekly IP docetaxel demonstrated a low toxicity and high efficacy for peritoneal carcinomatosis with dual anti-cancer effects via the peritoneal surface and capillary blood supply due to its unique pharmacokinetic property. PMID- 18425394 TI - Transient suppression of nuclear Cdc2 activity in response to ionizing radiation. AB - Suppression of Cdc2 activity is essential for DNA damage-induced G2 arrest. In the present study, we elucidated regulatory mechanism of Cdc2 activity during radiation-induced transient G2 arrest. Exposure of the cells to gamma-radiation (4 Gy) led to a transient increase of cells in G2 at 12 h rather than M phase and then the cells resumed cell cycle progression from the G2 arrest. However, the levels of cyclin B1 and Cdc2 activity were increased in the whole cell extracts at 12 h. Despite cyclin B1 induction and increased level of Cdc2 activity after irradiation the activities in the nuclear fractions were transiently decreased at 12 h and returned to control levels by 24-48 h, demonstrating transient inhibition of nuclear translocation of cyclin B1 in response to radiation. Moreover, inhibitory phosphorylation of the Cdc2 on Tyr15 and the Cdc25C on Ser216 were increased concomitant with transient G2 arrest. The level of phosphorylated Wee1 and its activity were also markedly increased at 12 h after irradiation. In addition, radiation caused nuclear accumulation of p21(CIP1/WAF1) at 12 h, resulting in increased-binding of p21(CIP1/WAF1) to Cdc2. Nuclear p21(CIP1/WAF1) protein level and its binding to Cdc2 gradually returned to control level when the cells resumed cell cycle progression. However, total protein level of p21(CIP1/WAF1) continued to increase until 48 h after irradiation. Collectively, these results indicate that the suppression of nuclear import of cyclin B1, the induction of Wee1 kinase activity, and the transient nuclear accumulation of p21(CIP1/WAF1) may play important roles in the transient cell cycle delay in response to ionizing radiation. PMID- 18425393 TI - Polymorphisms in CYP1B1, GSTM1, GSTT1 and GSTP1, and susceptibility to breast cancer. AB - Polymorphisms in the cytochrome P450 1B1 (CYP1B1) and glutathione S-transferase (GST) drug metabolic enzymes, which are responsible for metabolic activation/detoxification of estrogen and environmental carcinogens, were analyzed for their association with breast cancer risk in 541 cases and 635 controls from a North Carolina population. Each polymorphism, altering the catalytic function of their respective enzymes, was analyzed in Caucasian and African-American women. As reported in previous studies, individual polymorphisms did not significantly impact breast cancer risk in either Caucasian or African American women. However, African-American women exhibited a trend towards a protective effect when they had at least one CYP1B1 119S allele (OR=0.53; 95% CI=0.20-1.40) and increased risk for those women harboring at least one CYP1B1 432V allele (OR=5.52; 95% CI=0.50-61.37). Stratified analyses demonstrated significant interactions in younger (age < or =60) Caucasian women with the CYP1B1 119SS genotype (OR=3.09; 95% CI=1.22-7.84) and younger African-American women with the GSTT1 null genotype (OR=4.07; 95% CI=1.12-14.80). A notable trend was also found in Caucasian women with a history of smoking and at least one valine allele at GSTP1 114 (OR=2.12; 95% CI=1.02-4.41). In Caucasian women, the combined GSTP1 105IV/VV and CYP1B1 119AA genotypes resulted in a near 2-fold increase in risk (OR=1.96; 95% CI=1.04-3.72) and the three way combination of GSTP1 105IV/VV, CYP1B1 119AS/SS and GSTT1 null genotypes resulted in an almost 4 fold increase in risk (OR=3.97; 95% CI=1.27-12.40). These results suggest the importance of estrogen/carcinogen metabolic enzymes in the etiology of breast cancer, especially in women before the age of 60, as well as preventative measures such as smoking cessation. PMID- 18425395 TI - The role of nitric oxide on DNA damage induced by benzene metabolites. AB - Benzene, a tobacco constituent, is a leukemogen in humans and a carcinogen in rodents. Several benzene metabolites generate superoxide anion (O(2)(.-)) and induce nitric oxide synthase in the bone marrow of mice. We hypothesized that the reaction of nitric oxide (*NO) with O(2)(.-) leads to the formation of peroxynitrite as an intermediate during benzene metabolism. This hypothesis was supported by demonstrating that the exposure of mice to benzene produced nitrated metabolites and enhanced the levels of protein-bound 3-nitrotyrosine in the bone marrow of mice in vivo. In the current study, we investigated the influence of nitric oxide, generated from sodium 1-(N,N-diethylamino)diazen-1-ium-1,2-diolate, on DNA strand breaks induced by each single or binary benzene metabolite at different doses and compared the levels of the DNA damage induced by each benzene metabolite in the presence of nitric oxide with the levels of DNA strand breaks induced by peroxynitrite at similar doses in vitro. We found that among benzene metabolites only 1,2,4-trihydroxybenzene (BT) can induce significant DNA damage in the absence of nitric oxide. While 1,4-dihydroxybenzene (HQ), 1,4-benzoquinone (BQ) and 1,2-dihydroxybenzene (CAT) require .NO to induce DNA strand breaks, hydroquinone was the most potent DNA-damaging benzene metabolite in the presence of *NO. The order of DNA breaks by benzene metabolites in the presence of *NO is: Peroxynitrite = HQ > BT > BQ > CAT. The *NO and O(2)(.-) scavengers inhibited DNA damage induced by [HQ+*NO]. Benzene, trans,trans-muconaldehyde, and phenol, do not induce DNA strand breaks either in the absence or presence of *NO. However, adding phenol to [HQ+*NO] leads to greater DNA damage than [HQ+*NO] alone. Collectively, these results suggest that nitric oxide is an important factor in DNA damage induced by certain benzene metabolites, probably via the formation of the peroxynitrite intermediate. Phenol, the major benzene metabolite that does not induce DNA damage alone and is inactive in vivo, synergistically enhances DNA damage induced by potent benzene metabolite in the presence of nitric oxide. PMID- 18425396 TI - Promoter methylation of death-associated protein kinase and its role in irradiation response in cervical cancer. AB - This study was aimed at investigating the death-associated protein kinase (DAPK) promoter methylation and its clinical relevance in cervical cancer. The DAPK promoter methylation was detected by methylation-specific PCR (MSP) and correlated with DAPK mRNA and protein expression. The effect of DAPK expression on the radiosensitivity of the cervical cancer cell line was assessed by overexpressing DAPK in the radioresistant cell line SiHa. DAPK hypermethylation was found in 56.08% of the cervical cancer samples and was associated with the tumor histological cell type of squamous cell carcinoma (p=0.002) and advanced tumor stage (p=0.005). Subsequently, DAPK protein expression was found to significantly decrease in cervical cancer samples when compared to normal tissues. The DAPK mRNA and protein expression levels were absent or remarkably reduced in SiHa and HeLa in which the DAPK promoter was hypermethylated. The expression levels of DAPK could be restored after demethylation treatment with 5 aza-2'-deoxycytidine. Overexpressing DAPK in vitro had no significant influence to the survival of the radioresistant SiHa cell after being challenged by irradiation. Our findings suggest that DAPK might not directly be responsible for the cellular radiosensitivity, however, DAPK hypermethylation appeared to be of prognostic significance in the advanced stages of cervical cancer. PMID- 18425397 TI - Her-2/neu amplification and breast cancer survival: results from the Shanghai breast cancer study. AB - Her-2/neu is a member of the epidermal growth factor receptor family that has been found to be overexpressed or amplified in approximately 20-30% of breast cancers. Negative prognosticators and a shortened survival have been shown to be associated with these changes in Her-2/neu, but previous studies have consisted of predominantly Caucasian populations. Additionally, chromogenic in situ hybridization (CISH) has been suggested to be a potential alternative to fluorescent in situ hybridization (FISH), the expensive and labor-intensive gold standard assay currently used for Her-2/neu amplification. This study evaluated breast cancer samples from 313 Chinese women participating in the Shanghai breast cancer study, of which 100 (32%) were found to have Her-2/neu amplification by either FISH or CISH methodologies. After a mean follow-up period of 6.67 years, Her-2/neu amplification was found to be significantly associated with an increased hazard of death, regardless of which assay was used to detect amplification. Patients with Her-2/neu amplification were approximately 60% more likely to die of the disease (HR: 1.6, 95% CI: 1.0-2.6) than patients without amplification, even after adjusting for age, stage, menopausal status, chemotherapy, radiotherapy and tamoxifen treatment. Furthermore, the negative prognostic effect of Her-2/neu varied by cancer stage, with greater risks of death evident among later stage patients. This study supports a negative prognostic role for Her-2/neu in breast cancer survival among a Chinese population, irrespective of whether FISH or CISH is used to detect amplification of the Her-2/neu gene. PMID- 18425398 TI - Adjuvant chemotherapy with tegafur/uracil administration after transcatheter arterial chemoembolization for advanced hepatocellular carcinoma. AB - Although transcatheter arterial chemoembolization (TACE) is considered to be an effective treatment for advanced hepatocellular carcinoma (HCC), it is difficult to achieve complete necrosis by TACE alone due to incomplete embolization and tumor angiogenesis. Recent studies have shown that tegafur/uracil (UFT) inhibits tumor angiogenesis in several cancer types. Therefore, this study was conducted to test the efficacy and toxicity of the UFT administration after TACE in advanced HCC. Thirty patients with HCC who had been treated with TACE alone more than three times and had a recurrence within 6 months were enrolled. All of the patients were treated with TACE and 28 patients were randomly assigned to the UFT (UFT 300 mg/day, three days after TACE, n=14) and control groups (n=14). The primary end point was the time to treatment failure (TTF) and the secondary end points were mainly the response rate and toxicity. Administration and observation were continued up to 6 months after TACE unless local recurrence was detected or serious adverse events developed. The median TTF in the control group was 87 days, whereas in the UFT group it was 127 days, thus significantly prolonged as compared to the control group (P=0.0016). Moreover, the overall response rate (35.7%) in the UFT group was significantly higher than that in the control group (0%). As for toxicity, only 4 patients in the UFT group developed grade 1-2 toxicities such as ascites. Serious complications by TACE were not observed in either group. Notably, there were no increases in the serum VEGF levels in the UFT group whereas those in the control group increased significantly. In conclusion, UFT administration after TACE was an effective treatment and showed no severe adverse events. This regimen may have an adjuvant role and antiangiogenic function in advanced HCC. PMID- 18425399 TI - Contaminant levels in rainbow trout, oncorhynchus mykiss, and their diets from Missouri coldwater hatcheries. AB - Organochlorine and metal contaminants often occur in commercial fish diets and can accumulate in fish to levels of concern for human consumption. Contaminant levels were investigated in diet and rainbow trout fillets from Missouri coldwater hatcheries used in "put and take" fisheries. The average fillet:diet ratio was <0.1 for lead and cadmium, 0.4-0.6 for organochlorine compounds, and about 0.8 for mercury. Trout fillet concentrations for all contaminants were low (<50 ng/g) and below Missouri's fish consumption advisory trigger levels. PMID- 18425400 TI - Midurethral sling outcomes: tension-free vaginal tape versus Pelvilace. AB - We sought to compare the complications and efficacy of tension-free vaginal tape (TVT) and Pelvilace. Chart reviews and telephone questionnaires documented complications, urinary symptoms [Urogenital Distress Inventory short form (UDI-6) and Patient Global Impression of Improvement (PGII)], and re-operations. Failure was defined as PGII > or = 4 (no change or worse) or reoperation for urinary incontinence. There were 91 TVT and 22 Pelvilace slings. The TVT group had lower rates of urinary retention beyond 1 week (8% vs. 43%, p < 0.001) and reoperation for retention (1% vs. 19%, p < 0.001). Median changes in UDI-6 scores were -33.3 for the TVT group and -29.6 for the Pelvilace group (p = 0.21). Median PGII scores were 1 for the TVT group and 2 for the Pelvilace group (p = 0.024). Ten percent of the TVT group and 24% of the Pelvilace group (p = 0.095) were failures. Urinary retention was more common after Pelvilace than TVT slings. Symptomatic outcomes and failure rates were similar. PMID- 18425401 TI - The impact of multichannel urodynamics upon treatment recommendations for female urinary incontinence. AB - The aim of this study was to evaluate whether multichannel urodynamic testing changes a physician's treatment recommendations when managing women with urinary incontinence. In this prospective reader study, four fellowship-trained urogynecologists reviewed 39 abstracted cases of urinary incontinence on two occasions: first without and subsequently with urodynamic data. Treatment recommendations were made for each case after each review. The probability of urodynamic data modifying treatment recommendations was estimated for each reader and for the population of readers using a random effects logistic regression to account for reader variability. The overall probability that urodynamic data would change treatment was 26.9% (95% confidence interval (CI), 18.6%, 37.2%) for medical treatments and 45.5% (95% CI, 37.8%, 53.4%) for surgical treatments. Reader-to-reader differences accounted for 3% and <1% of the total variance for medical and surgical treatments, respectively. Multichannel urodynamic evaluations are significantly associated with changes in medical and surgical treatment recommendations in a referral population. PMID- 18425402 TI - Effects of three years of low-dose thiazides on mineral metabolism in healthy elderly persons. AB - In this clinical trial of 12.5 or 25 mg/day of hydrochlorothiazide, the urine calcium showed significant decreases from placebo in men at one year, but the effects had waned by 3 years. Serum bicarbonate was consistently greater in the thiazide than in the placebo groups throughout the three years. These effects could be beneficial to the skeleton. INTRODUCTION: Previous studies have shown increased bone density and reduced risk of fracture in patients taking thiazide diuretics. The long-term effects of low-dose thiazides on mineral metabolism have not been reported in normal subjects. METHODS: We conducted a randomized, double blinded trial in normals aged 60-79 years, using hydrochlorothiazide 12.5 or 25 mg/d or placebo for three years. Subjects were encouraged to maintain calcium intake of 1,000 to 1,500 mg/day. Measurements of serum and urine calcium metabolism were done at baseline, six months, and yearly. Data were analyzed in 88 men and 177 women who had taken study medication. Adjusted change in the measurements from baseline to one and three years were compared among groups. RESULTS: The calcium intake increased in all groups. Urine calcium per day was significantly lower in thiazide than placebo groups in men at one year but not at three years; in women the changes were not significantly different. Serum bicarbonate was higher in thiazide compared to placebo groups at one and three years. No changes were seen in serum calcium, phosphate, parathyroid hormone, sodium or magnesium. CONCLUSIONS: The results suggest that both increased calcium availability from a hypocalciuric effect and reduction in acid-induced bone buffering could be mechanisms for the beneficial skeletal effects. PMID- 18425404 TI - DPROT: prediction of disordered proteins using evolutionary information. AB - The association of structurally disordered proteins with a number of diseases has engendered enormous interest and therefore demands a prediction method that would facilitate their expeditious study at molecular level. The present study describes the development of a computational method for predicting disordered proteins using sequence and profile compositions as input features for the training of SVM models. First, we developed the amino acid and dipeptide compositions based SVM modules which yielded sensitivities of 75.6 and 73.2% along with Matthew's Correlation Coefficient (MCC) values of 0.75 and 0.60, respectively. In addition, the use of predicted secondary structure content (coil, sheet and helices) in the form of composition values attained a sensitivity of 76.8% and MCC value of 0.77. Finally, the training of SVM models using evolutionary information hidden in the multiple sequence alignment profile improved the prediction performance by achieving a sensitivity value of 78% and MCC of 0.78. Furthermore, when evaluated on an independent dataset of partially disordered proteins, the same SVM module provided a correct prediction rate of 86.6%. Based on the above study, a web server ("DPROT") was developed for the prediction of disordered proteins, which is available at http://www.imtech.res.in/raghava/dprot/. PMID- 18425403 TI - Weight-bearing bones are more sensitive to physical exercise in boys than in girls during pre- and early puberty: a cross-sectional study. AB - We carried out a cross-section study of the sex-specific relationship between bone mineral content and physical activity at sites with different loading in pre and early pubertal girls and boys. There was significant sensitivity of bone mineral content of the hip to physical exercise in boys, but not in girls. BACKGROUND: Since little is known whether there are sex differences in sensitivity of bone to loading, we investigated sex differences in the cross sectional association between measures of physical activity (PA) and bone mass and size in pre- and early pubertal children of both sexes. METHODS: We measured bone mineral content/density (BMC/BMD) and fat-free mass (FFM) in 269 6- to 13 year-old children from randomly selected schools by dual-energy X-ray absorptiometry. Physical activity (PA) was measured by accelerometers and lower extremity strength by a jump-and-reach test. RESULTS: Boys (n = 128) had higher hip and total body BMC and BMD, higher FFM, higher muscle strength and were more physically active than girls (n = 141). Total hip BMC was positively associated with time spent in total and vigorous PA in boys (r = 0.20-0.33, p < 0.01), but not in girls (r = 0.02-0.04, p = ns), even after adjusting for FFM and strength. While boys and girls in the lowest tertile of vigorous PA (22 min/day) did not differ in hip BMC (15.62 vs 15.52 g), boys in the highest tertile (72 min/day) had significantly higher values than the corresponding girls (16.84 vs 15.71 g, p < 0.05). CONCLUSIONS: Sex differences in BMC during pre- and early puberty may be related to a different sensitivity of bone to physical loading, irrespective of muscle mass. PMID- 18425405 TI - Multi-agent-based bio-network for systems biology: protein-protein interaction network as an example. AB - Recently, a collective effort from multiple research areas has been made to understand biological systems at the system level. This research requires the ability to simulate particular biological systems as cells, organs, organisms, and communities. In this paper, a novel bio-network simulation platform is proposed for system biology studies by combining agent approaches. We consider a biological system as a set of active computational components interacting with each other and with an external environment. Then, we propose a bio-network platform for simulating the behaviors of biological systems and modelling them in terms of bio-entities and society-entities. As a demonstration, we discuss how a protein-protein interaction (PPI) network can be seen as a society of autonomous interactive components. From interactions among small PPI networks, a large PPI network can emerge that has a remarkable ability to accomplish a complex function or task. We also simulate the evolution of the PPI networks by using the bio operators of the bio-entities. Based on the proposed approach, various simulators with different functions can be embedded in the simulation platform, and further research can be done from design to development, including complexity validation of the biological system. PMID- 18425406 TI - Protein-osmolality ratio for quantification of proteinuria in children. AB - Proteinuria is an important factor for renal injury and prognosis in many diseases. The most valuable method for evaluation of proteinuria is quantitative protein analysis in appropriately 24-h collected urine. But urine collection is difficult and cumbersome especially in children and working adults. The aim of the present study is to define the usefulness of urinary protein/urine osmolality (Uprot/Uosm) ratio in quantification of proteinuria. One hundred and seventy-one patients whose age ranged between 3 and 14 years were included in the study. Uprot/Uosm (r = 0.85, P < 0.001) and urinary protein/creatinine Uprot/Ucrea (r = 0.81 and P < 0.001) ratios were significantly correlated with 24-h protein excretion. Twenty-four-hour protein excretion was correlated with Uprot/Ucrea (r = 0.76, P < 0.001) and Uprot/Uosm (r = 0.79, P < 0.001) ratios in proteinuric group. But, there was no correlation between Uprot/Uosm and Uprot/Ucrea ratios with 24-h protein excretion in non-proteinuric group. The positive and negative predictive value of Uprot/Uosm ratio of 0.28 mg/l/mOsm/kg was 89.9 and 90.8% and Uprot/Ucrea ratio of 0.24 mg/mg was 85.7 and 90% for estimating proteinuria (above 4 mg/m2 h(-1)). The 95th percentile of Uprot/Uosm ratio was 0.25 mg/l/mOsm/kg and Uprot/Ucrea ratio was 0.27 mg/mg in normal group. The best estimate was Uprot/Uosm ratio of 1.42 (sensitivity 100%, specificity 94.9%) and Uprot/Ucrea ratio of 0.75 (sensitivity 100%, specificity 92.9%) for nephrotic proteinuria (40 mg/m2 h(-1)). Uprot/Uosm which is a reliable and simple method can be used for quantification of proteinuria in pediatric patients with normal renal function. Using Uprot/Uosm ratio for quantification of proteinuria can remove the necessity of 24-h urine collection, urine creatinine measurement and spending additional cost and time if the center measures the urine osmolality routinely in urine analysis. PMID- 18425407 TI - Probing the material properties and phase transitions of ferroelectric liquid crystals by determination of the Landau potential. AB - The full Landau potential of several, widely varying ferroelectric liquid crystalline materials has been experimentally determined. Tilt angle and polarisation data is analysed across the SmA to SmC transition for varying applied electric-field amplitudes, allowing the determination of all the coefficients of the generalised Landau model of ferroelectric liquid crystals. The materials investigated encompass different materials, including low polarisation mixtures to high-polarisation single-component materials. The materials also possess a variation in the order of the SmA to SmC phase transition from strongly first order to strongly second order. The effects of both the polarisation and order of phase transition of the system are discussed with respect to the various terms of the generalised Landau model. Further, the mechanisms behind the difference between a first- and second-order phase transition are discussed with respect to the Landau potential and the second Landau coefficient b . PMID- 18425408 TI - Tunable-slip boundaries for coarse-grained simulations of fluid flow. AB - On the micro- and nanoscale, classical hydrodynamic boundary conditions such as the no-slip condition no longer apply. Instead, the flow profiles exhibit "slip" at the surface, which is characterized by a finite slip length (partial slip). We present a new, systematic way of implementing partial-slip boundary conditions with arbitrary slip length in coarse-grained computer simulations. The main idea is to represent the complex microscopic interface structure by a spatially varying effective viscous force. An analytical equation for the resulting slip length can be derived for planar and for curved surfaces. The comparison with computer simulations of a DPD (dissipative particle dynamics) fluid shows that this expression is valid from full slip to no slip. PMID- 18425409 TI - Multistage polar switching in bent-core mesogens. AB - We report a novel type of electro-optical switching in a tilted smectic phase of bent-shaped mesogens. The switching consists of a continuous stage and two bistable transitions. Detailed optical and electro-optical measurements using high-speed imaging are given and possible interpretations of the experimental results are discussed. PMID- 18425410 TI - Characterization of single semiflexible filaments under geometric constraints. AB - Confinement effects on single semiflexible macromolecules are of central importance for a fundamental understanding of cellular processes involving biomacromolecules. To analyze the influence of confinement on the fluctuations of semiflexible macromolecules we study individual actin filaments in straight and curved microchannels. We experimentally characterize the segment distributions for fluctuating semiflexible filaments in microchannels as a function of the channel width. Moreover, the effect of channel curvature on the filament fluctuations is investigated. We find quantitative agreement between experimental results, Monte Carlo simulations, and the analytical description. This allows for determination of the persistence length of actin filaments, the deflection length, which characterizes the confinement effects, and the scaling exponents for the segment distribution of semiflexible macromolecules. PMID- 18425411 TI - Highly specific interactions between botulinum neurotoxins and synaptic vesicle proteins. AB - Despite its extreme toxicity, botulinum neurotoxin is widely utilized in low doses as a treatment for several neurological disorders; higher doses cause the neuroparalytic syndrome botulism. The toxin blocks neurotransmitter release by preferentially attaching to pre-synaptic membrane receptors at neuromuscular junctions and subsequently delivering a Zn2+-dependent protease component to presynaptic neuronal cytosol. These highly specialized enzymes exclusively hydrolyze peptide bonds within SNARE (soluble N-ethylmaleiamide sensitive factor attachment protein receptor) proteins. In this review we discuss the structural basis for botulinum toxin's exquisite specificity for its neuronal cell-surface receptors and intracellular SNARE targets. PMID- 18425412 TI - Myostatin as a therapeutic target for musculoskeletal disease. PMID- 18425413 TI - Triple arginines in the cytoplasmic tail of endomannosidase are not essential for type II membrane topology and Golgi localization. AB - Endomannosidase is a Golgi-localized endoglycosidase, which provides an alternate glucosidase-independent pathway of glucose trimming. Using a protease protection assay we demonstrated that Golgi-endomannosidase is a type II membrane protein. The first 25 amino acids of this protein, containing the cytoplasmic tail and the transmembrane domain, were sufficient for Golgi retention of fused reporter proteins alpha1-antitrypsin or green fluorescent protein. However, shortening or deletion of the transmembrane domain prevented Golgi localization, while lengthening it partially reduced Golgi retention of the enzyme. Substitution of the highly conserved positively charged amino acids within the cytoplasmic tail had neither an effect on type II topology nor on the inherent Golgi localization of the enzyme. In contrast, cytoplasmic tail-deleted rat endomannosidase possessed an inverted topology resulting in endoplasmic reticulum mislocalization. Thus, proper topology rather than the presence of positively charged amino acids in the cytoplasmic tail is critical for Golgi localization of rat endomannosidase. PMID- 18425414 TI - Cardiolipin, the heart of mitochondrial metabolism. AB - Cardiolipin is a unique phospholipid, which is almost exclusively localized in the mitochondrial inner membrane where it is synthesized from phosphatidylglycerol and cytidinediphosphate-diacylglycerol. After primary synthesis, the mature acyl chain composition of cardiolipin is achieved by at least two remodeling mechanisms. In the mitochondrial membrane cardiolipin plays an important role in energy metabolism, mainly by providing stability for the individual enzymes and enzyme complexes involved in energy production. Moreover, cardiolipin is involved in different stages of the mitochondrial apoptotic process and in mitochondrial membrane dynamics. Cardiolipin alterations have been described in various pathological conditions. Patients suffering from Barth syndrome have an altered cardiolipin homeostasis caused by a primary deficiency in cardiolipin remodeling. Alterations in cardiolipin content or composition have also been reported in more frequent diseases such as diabetes and heart failure. In this review we provide an overview of cardiolipin metabolism, function and its role in different pathological states. PMID- 18425415 TI - Dissection of the molecular mechanisms that control the nuclear accumulation of transport factors importin-alpha and CAS in stressed cells. AB - The physiological state of eukaryotic cells controls nuclear trafficking of numerous cargos. For example, stress results in the inhibition of classical protein import, which is characterized by the redistribution of several transport factors. As such, importin-alpha and cellular apoptosis susceptibility protein (CAS) accumulate in nuclei of heat-shocked cells; however, the mechanisms underlying this relocation are not fully understood. We now show that heat upregulates the initial docking of importin-alpha at the nuclear envelope and stimulates the translocation of CAS into the nuclear interior. Moreover, heat exposure compromises the exit of importin-alpha from nuclei and drastically increases its retention in the nucleoplasm, whereas CAS nuclear exit and retention are less affected. Taken together, our results support the idea that heat shock regulates importin-alpha and CAS nuclear accumulation at several levels. The combination of different stress-induced changes leads to the nuclear concentration of both transport factors in heat-stressed cells. PMID- 18425416 TI - Heparanase involvement in physiology and disease. AB - Heparanase is an endoglycosidase that degrades heparan sulfate on the cell surface and extracellular matrix. The physiological functions of heparanase include heparan sulfate turnover, embryo development, hair growth, and wound healing. Heparanase is implicated in a variety of pathologies, such as tumor growth, angiogenesis, metastasis, inflammation, and glomerular diseases. Heparanase overexpression in a variety of malignant tumors suggests that it could be a target for anti-cancer therapy. PMID- 18425417 TI - Considerations on temperature, longevity and aging. AB - A modest reduction in body temperature prolongs longevity and may retard aging in both poikilotherm and homeotherm animals. Some of the possible mechanisms mediating these effects are considered here with respect to major aging models and theories. PMID- 18425418 TI - Early phase II trial of oral vorinostat in relapsed or refractory breast, colorectal, or non-small cell lung cancer. AB - Vorinostat (Zolinza) is a histone deacetylase inhibitor that has demonstrated activity in patients with advanced solid tumors in phase I trials. A multicenter, open-label phase II trial of oral vorinostat 200, 300 or 400 mg bid for 14 days followed by a 7-day rest until disease progression or intolerable toxicity was conducted. Patients with measurable, relapsed or refractory breast or non-small cell lung cancer who had received > or = 1 prior therapy or colorectal cancer who had received > or = 2 prior therapies were eligible. The response rate, safety and tolerability were evaluated. Sixteen patients (median age, 62 years; median 5.5 prior therapies) were enrolled. Six patients received 400 mg bid, six received 300 mg bid and four received 200 mg bid (14 days/3 weeks). Dose-limiting toxicities (DLTs) at the 400 or 300 mg bid levels were anorexia, asthenia, nausea, thrombocytopenia, vomiting, and weight loss. No DLTs were observed at the 200 mg bid level. Disease stabilization was observed in eight patients, but there were no confirmed responses. The median TTP was 33.5 days. Eleven patients discontinued due to clinical adverse experiences (AEs). The most common drug related AEs were anorexia (81%), fatigue (62%), nausea (62%), diarrhea (56%), vomiting (56%), thrombocytopenia (50%) and weight loss (50%). Drug-related AEs > or = grade 3 included thrombocytopenia (50%), anemia (12%), asthenia (12%) and nausea (12%). Vorinostat in a daily oral schedule for 14 days/3 weeks was tolerable at 200 mg bid only, and no responses were observed in this study. Most patients, however, had limited drug exposure which did not allow a reliable efficacy analysis. PMID- 18425419 TI - A phase I and pharmacokinetic study of XK469R (NSC 698215), a quinoxaline phenoxypropionic acid derivative, in patients with refractory acute leukemia. AB - A phase I study was performed to determine the safety and pharmacokinetics of XK469R in patients with refractory acute leukemia. The study aimed to determine the maximum tolerated dose (MTD) and dose limiting toxicity (DLT) of XK469R given intravenously over 30 to 60 min on days 1, 3, and 5 of a 21 day cycle. Patients were treated in successive cohorts of six until DLT was observed. Once the MTD was determined, an additional cohort of six patients was enrolled at the previous dose level and that dose was considered the recommended phase 2 dose (RPTD). Forty-six patients were treated at dose levels of 1,400, 1,750, 2,200, and 2,750 mg. The DLTs were: mucositis, colitis and hyperbilirubinemia. Reversible myelosuppression was noted at all dose levels. One (2%) of 42 patients achieved a complete remission and five patients (11%) had hematologic improvement. The half life of the drug was long with a mean value of 48 h. The mean clearance was 206 mL/h with a coefficient of variation of 32%. No correlation was observed between the development of DLT and pharmacokinetics. The RTPD is 1,750 mg. XK469R induced hematological responses in patients with refractory leukemia at tolerable doses. PMID- 18425420 TI - Effect of telmisartan on cardiovascular complications associated with streptozotocin diabetic rats. AB - Angiotensin receptor blockers provide cardiovascular protection in heart failure patients. We have studied the effect of 8 weeks treatment with telmisartan (5 mg kg(-1) day(-1)) on cardiovascular complications associated with streptozotocin (STZ) diabetic rats. Wistar rats were made diabetic with STZ (45 mg kg(-1), iv). Various biochemical and cardiac parameters were measured at the end of 8 weeks. STZ produced hyperglycemia, hypoinsulinemia, hyperlipidemia, increased blood pressure, increased creatinine, cardiac enzyme and C-reactive protein (CRP) levels, reduction in heart rate and cardiac hypertrophy. Chronic treatment with telmisartan significantly (P < 0.05) prevented STZ induced hypertension and elevated fasting glucose level with simultaneous increase in serum insulin levels. It significantly (P < 0.05) reduced the elevated cholesterol, very low density lipoprotein (VLDL) and triglyceride levels in diabetic rats and increased the lower high density lipoprotein (HDL)-cholesterol levels. Further, telmisartan produced a significant (P < 0.05) reduction in the elevated creatinine levels, CRP and levels of other cardiac enzyme markers like Lactate de-hydrogenase and creatinine kinase of diabetic rats. STZ-induced bradycardia was also prevented by telmisartan treatment and it also produced beneficial effect by preventing cardiac hypertrophy as evident from left ventricular collagen levels, cardiac hypertrophy index and left ventricular hypertrophy index of diabetic rats. Our data suggest that telmisartan prevents not only the STZ-induced metabolic abnormalities, but also cardiovascular complications. PMID- 18425421 TI - Proapoptotic activity of cytochrome c in living cells: effect of K72 substitutions and species differences. AB - Cytochrome c is one of the key proteins involved in the programmed cell death, and lysine 72 is known to be required for its apoptogenic activity. We have engineered a number of horse and murine cytochrome c single-point mutants with various substitutions at position 72 and compared quantitatively their proapoptotic activity in living cells. Apoptosis was activated by transferring exogenous cytochrome c into the cytoplasm of cells via a nontraumatic electroporation procedure. All mutant proteins studied exhibited significantly reduced proapoptotic activities in comparison with those for the wild type cytochromes. Relative activity of the horse (h(K72X)) and murine (m(K72W)) mutant proteins diminished in the order: h(K72R) > h(K72G) > h(K72A) > h(K72E) > h(K72L) >> h(K72W) > m(K72W). As estimated, the horse and murine K72W mutants were at least 200- and 500-fold less active than corresponding wild type proteins. Thus, the K72W-substituted cytochrome c can serve as an adequate candidate for knock-in studies of cytochrome c-mediated apoptosis. The proapoptotic activity of wild type cytochrome c from different species in murine monocytic WEHI-3 cells reduced in the order: murine cytochrome c > human cytochrome c approximately horse cytochrome c, thus indicating that apoptotic effect of cytochrome c depends on the species compatibility. PMID- 18425422 TI - Proteome analysis of the transformation potential of the Epstein-Barr virus encoded latent membrane protein 1 in nasopharyngeal epithelial cells NP69. AB - Latent membrane protein 1 (LMP1) of Epstein-Barr virus has been identified to be crucial in inducing cell transformation. However, the mechanism of LMP1-mediated epithelial cell transformation remains unclear. In this study, nasopharyngeal epithelial cells NP69 were infected with retrovirus with gene encoding wild type LMP1 or mutational LMP1 defective in binding to tumor necrosis factor receptor associated death domain (TRADD). The NP69-LMP1(TRADD) lost some malignant phenotypes compared with the NP69-LMP1(WT). We performed proteomic approach to gain the differential protein expression profile associated with LMP1-mediated epithelial cell transformation. Furthermore, the differential expressional levels of partial identified proteins were confirmed by Western blot and real-time RT PCR. Some were known to be related to the development of LMP1-induced transformation, and some were new LMP1-associated proteins. These data are valuable for further study of the mechanism of LMP1 in human nasopharyngeal carcinoma and provide some new clues for investigating other LMP1-associated tumors. PMID- 18425423 TI - Chemopreventive activity of lantadenes on two-stage carcinogenesis model in Swiss albino mice: AP-1 (c-jun), NFkappaB (p65) and P53 expression by ELISA and immunohistochemical localization. AB - Lantadenes are pentacyclic triterpenoids isolated from leaves of Lantana camara L. and have antitumor activity. Lantadene A (LA) and methyl ester of LA (LAM) were earlier studied in the author's lab for their chemopreventive effect on 7,12 dimethylbenz(a)anthracene (DMBA) followed by 12-O-tetradecanoylphorbol-13-acetate (TPA) induced squamous cell carcinoma incidence in Swiss albino mice. The present study was specially designed to initiate the involvement of the molecular targets in chemopreventive activity of these compounds. Skin lesions were induced by twice-weekly topical application of DMBA (100 nmol/100 microl of acetone) for 2 weeks followed by TPA (1.7 nmol/100 microl of acetone) on depilated back of mice for 20 weeks. LA and LAM were administered orally at a dose of 50 mg/kg body weight twice weekly, 1 week before DMBA application and continued for 20 weeks thereafter. A significant decrease in the incidence of number of lesions in mice was obtained in LA/LAM treated groups as compared to DMBA/TPA alone. Significant increase in the protein levels of c-jun, p65, and p53 by ELISA were observed in DMBA/TPA treated mice tumors whereas less expression was observed in LA and LAM treated tumors. Further immunohistochemical localization of transcription factors was studied which also showed less localization of c-jun, p65, and p53 in LA and LAM treated tumors as compared to localization in DMBA/TPA treated tumors. It can be inferred that LA and LAM chemopreventive activity may be linked to the deregulation of above molecular targets which warrants further studies in that direction. PMID- 18425424 TI - Biosynthesis, processing, trafficking, and enzymatic activity of mouse neprilysin 2. AB - Neprilysin 2 (NEP2) has been recently identified as a new member of the M13 subfamily of zinc-dependent metalloproteases and shares a highly homologous amino acid sequence with neprilysin (EC 3.4.24.11, NEP). NEP2 has been reported to exist as membrane-bound and soluble secreted variants. To investigate mechanisms of regulating NEP2 activity, we developed a simple and sensitive method for measuring NEP2 activity using synthetic substrates with a fluorescent probe. NEP2 only cleaved Suc-Ala-Ala-Phe-AMC, while NEP cleaved both Dansyl-D-Ala-Gly-p-nitro Phe-Gly and Suc-Ala-Ala-Phe-AMC. Using HEK293 cells stably expressing mouse NEP2, we evaluated the effects of various reagents affecting post-translational modification and protein trafficking on extracellular NEP2 activity secreted into the culture medium. Inhibition of N-glycosylation by tunicamycin reduced both the enzymatic activity of extracellular NEP2 and the molecular size of intracellular NEP2. Disruption of the Golgi apparatus with brefeldin A markedly reduced extracellular NEP2 activity in parallel with intracellular NEP2 protein level in HEK293 cells. In contrast, the cytoskeleton disrupting reagents, nocodazole and cytochalasin B barely affected NEP2 activity. Two distinct calcium-perturbing reagents, a calcium ionophore A23187 and thapsigargin, reduced extracellular NEP2 activity. However, A23187-mediated down-regulation was not rescued by co treatment with inhibitors of MAPK, calmodulin, or the proteasome/calpains. In conclusion, we established a simple and sensitive protocol which was able to discriminate NEP2 and NEP activity, and showed that intracellular transport and secretion of NEP2 is regulated by processes such as glycosylation, ER-Golgi transport, and intracellular calcium levels. PMID- 18425426 TI - Continuous dechlorination of tetrachloroethene in an upflow anaerobic sludge blanket reactor. AB - Influences of hydraulic retention time (HRT) on dechlorination of tetrachloroethene (PCE) were investigated in an upflow anaerobic sludge blanket (UASB) reactor inoculated with anaerobic granular sludge non-pre-exposed to chlorinated compounds. PCE was introduced into the reactor at a loading rate of 3 mg/l d. PCE removal increased from 51+/-5% to 87+/-3% when HRT increased from 1 to 4 d, corresponding to an increase in the PCE biotransformation rate from 10.5+/-2.3 to 21.3+/-3.7 micromol/d. A higher ethene production rate, 0.9+/-0.2 micromol/d, was attained without accumulation of dichloroethenes at the HRT of 4 d. Dehalococcoides-like species were detected in sludge granules by fluorescence in situ hybridization, with signal strength in proportion to the extent of PCE dechlorination. PMID- 18425425 TI - The role of ErbB3 and its binding partners in breast cancer progression and resistance to hormone and tyrosine kinase directed therapies. AB - An increasingly important role for the ErbB3 receptor in the genesis and progression of breast cancer is emerging. ErbB3 is frequently overexpressed in breast cancer and coexpression of ErbB2/3 is a poor prognostic indicator. ErbB3 has also been implicated in the development of resistance to antiestrogens such as tamoxifen and ErbB tyrosine kinase inhibitors such as gefitinib. Persistent activation of the AKT pathway has been postulated to contribute to ErbB3-mediated resistance to these therapies. This activation may be due in part to the inappropriate production of the ErbB3 ligand heregulin. ErbB3 binding proteins, which negatively regulate ErbB3 protein levels and the ability of ErbB3 to transmit proliferative signals, also contribute to breast cancer progression and treatment resistance. These proteins include the intracellular RING finger E3 ubiquitin ligase Nrdp1 and the leucine-rich protein LRIG-1 that mediate receptor degradation. Ebp1, another ErbB3 binding protein, suppresses HRG driven breast cancer cell growth and contributes to tamoxifen sensitivity. These studies point to the importance of the evaluation of protein levels and functional activity of ErbB3 and its binding proteins in breast cancer prognosis and prediction of clinical response to treatment. PMID- 18425428 TI - Epoxide hydrolase-mediated enantioconvergent bioconversions to prepare chiral epoxides and alcohols. AB - A number of epoxide hydrolase (EH)-mediated bioconversions have been developed to prepare single enantiomeric product from racemic substrates with a yield greater than 50%. Enantioconvergent hydrolysis using single or two EHs possessing complementary enantio- and regio-selectivity, EH-based chemoenzymatic reactions, and EH-triggered cascade-reactions have been developed for the preparation of chiral epoxides, epoxyalcohols, tetrahydrofuran derivatives and vicinal diols. All these bioconversions are based on stereochemical flexibilities of various EHs and can be used in total synthesis of biologically active compounds without the formation of unwanted enantiomers. PMID- 18425427 TI - Physiological activities of a beta-glucan produced by Panebacillus polymyxa. AB - In vitro bioactivities of a beta-glucan produced by Panebacillus polymyxa JB115 were investigated. Nitric oxide production by RAW 264.7 macrophage cells pre treated with beta-glucan JB115 (from 0.1 to 1 mg ml(-1)) was significantly increased, compared to that in untreated cells (P < 0.001). The beta-glucan JB115 increased superoxide radical-scavenging activity by 66% at 1 mg ml(-1). It also suppressed hyaluronidase (32%) and collagenase (33%) activities and, additionally, displayed antitumor activity, blocking the growth of Sarcoma 180 cells in a concentration-dependent manner. The immune-stimulatory, antioxidant, collagenase inhibitory and hyaluronidase inhibitory effects of the beta-glucan support its potential role in the prevention of bacterial disease against fish and in the protection of skin against aging. PMID- 18425429 TI - Solid-liquid two-phase partitioning bioreactors for the treatment of gas-phase volatile organic carbons (VOCs) by a microbial consortium. AB - A two-phase partitioning bioreactor (TPPB), employing styrene-butadiene co polymer beads as the sequestering/delivery phase, was used to treat high step change loadings of toluene in a contaminated air stream. The polymers, which are biocompatible and non-bioavailable, allowed the use of a microbial consortium and effectively absorbed and released the toluene vapours for biodegradation, while providing a buffering effect against high toluene transients. Toluene loadings were increased from a base steady state rate of 343-6,000 g/m(3) h for 1 h periods, with the polymer-aqueous system substantially outperforming a single phase system on the basis of improving the toluene removal efficiency and reducing the maximum toluene concentrations emitted during the transients. PMID- 18425430 TI - Oral administration of recombinant human granulocyte-macrophage colony stimulating factor expressed in rice endosperm can increase leukocytes in mice. AB - Human granulocyte-macrophage colony stimulating factor (hGM-CSF) is used clinically to treat leucopenia typically caused by cancer chemotherapy or radiotherapy. This study used multiple strategies to obtain very high expression levels of OsrhGM-CSF (14 microg/seed) in rice endosperm. Electron micrographs of immunogold-labeled transgenic endosperm showed that rhGM-CSF was not only localized in protein bodies but was also distributed in the apoplast. A biological activity assay indicated that OsrhGM-CSF stimulated the growth of TF-1 cells in vitro. In addition, the transgene was used to effectively treat leucopenia by oral administration of the unprocessed transgenic grains. In cyclophosphamide-induced leucopenic mice, transgenic seeds produced a 27% (t=0.021) gain in leukocytes after 14 days feeding. Even in non-leucopenic mice, leukocyte gain was 37% (t=0.002) more than that of mice fed non-transgenic seeds. This study provides a novel approach to the use of oral unprocessed transgenic OsrhGM-CSF seeds to treat leucopenia. PMID- 18425431 TI - siRNA-mediated silencing of the 37/67-kDa high affinity laminin receptor in Hep3B cells induces apoptosis. AB - The laminin-binding protein, variously called the 37/67-kDa high affinity laminin receptor or p40, mediates the attachment of normal cells to the laminin network, and also has a role as a ribosomal protein. Over-expression of this protein has been strongly correlated with the metastatic phenotype. However, few studies have investigated the cellular consequence of the ablation of this gene's expression. To address this issue, the expression of the 37/67-kDa high affinity laminin receptor was knocked out with several siRNA constructs via RNA interference in transformed liver (Hep3B) cells. In each case where the message was specifically ablated, apoptosis was induced, as determined by annexin V/propidium iodide staining, and by double staining with annexin V and an antibody directed against the 37/67-kDa high affinity laminin receptor. These results suggest that this protein plays a critical role in maintaining cell viability. PMID- 18425432 TI - Effects of nanoanatase on the photosynthetic improvement of chloroplast damaged by linolenic acid. AB - To further evaluate the photosynthetic effects of nanoanatase, the improvement of spinach chloroplast photosynthesis damaged by linolenic acid was investigated in the present paper. Several results showed that after the addition of nanoanatase to the linolenic acid-treated chloroplast, the light absorption increased by linolenic acid could be decreased, but the excitation energy distribution from photosystem (PS) I to PS II was promoted, and the decrease of PS II fluorescence yield caused by linolenic acid was reduced and the inhibition of oxygen evolution caused by linolenic acid of several concentrations was decreased. It was considered that nanoanatase could combine with linolenic acid and decrease the damage of linolenic acid on the structure and function of chloroplast. PMID- 18425433 TI - The effect of peripheral administration of zinc on food intake in rats fed Zn adequate or Zn-deficient diets. AB - Zinc deficiency induces a striking reduction of food intake in animals. To elucidate the mechanisms for this effect, two studies were connectedly conducted to determine the effects of peripheral administration of zinc on food intake in rats fed the zinc-adequate or zinc-deficient diets for a 3-week period. In study 1, two groups of male Sprague-Dawley rats were provided diets made either adequate (ZA; 38.89 mg/kg) or deficient (ZD; 3.30 mg/kg) in zinc. In study 2, after feeding for 3 weeks, both ZA and ZD groups received intraperitoneal (IP) injection of zinc solution with three levels (0.5, 1.0, and 2.0 microg zinc/g body weight, respectively) and cumulative food intake at 0.5, 1, 2, 4, and 24 h, and plasma hormones concentrations were measured. The results in study 1 showed rats fed the ZD diets revealed symptoms of zinc deficiency, such as sparse and coarse hair, poor appetite, susceptibility to surroundings, lethargy, and small movements. Zinc concentrations in serum, femur, and skeletal muscle of rats fed the ZD diets declined by 26.58% (P < 0.01), 27.32% (P < 0.01), and 24.22% (P < 0.05), respectively, as compared with ZA control group. These findings demonstrated that rat models with zinc deficiency and zinc adequacy had been fully established. The results in study 2 showed that IP administration of zinc in both ZA and ZD rats did not influence food intake at each time points (P > 0.05), although zinc deficiency suppressed food intake. Plasma neuropeptide Y (NPY) was higher, but insulin and glucagon were lower in response to zinc deficiency or zinc administration by contrast with their respective controls (P < 0.05). Leptin, T3, and T4 concentrations were uniformly decreased (P < 0.05) in rats fed the ZD diets in contrast to ZA diets; however, no differences (P > 0.05) were observed during zinc injection. Calcitonin gene-related peptide was unaffected (P > 0.05) by either zinc deficiency or zinc administration. The present studies suggested that zinc administration did not affect short-term food intake in rats even in the zinc-deficient ones; the reduced food intake induced by zinc deficiency was probably associated with the depression in thyroid hormones. The results also indicated that NPY and insulin varied conversely during the control of food intake. PMID- 18425434 TI - Fat emboli unleashed: an exceptional etiology of encephalitis in sickle cell disease. PMID- 18425435 TI - Molecular physiology and pathophysiology of lysosomal membrane transporters. AB - In contrast to lysosomal hydrolytic enzymes, the lysosomal membrane remains poorly characterized. In particular, although the genetic study of cystinosis and sialic acid storage disorders led to the identification of two lysosomal transporters for cystine and sialic acids, respectively, ten years ago, most transporters responsible for exporting lysosomal hydrolysis products to the cytosol are still unknown at the molecular level. However, two lines of investigation recently started to fill this gap in the knowledge of lysosomal biology. First, novel proteomic approaches are now able to provide a reliable inventory of lysosomal membrane proteins. On the other hand, a novel functional approach based on intracellular trafficking mechanisms allows direct transport measurement in whole cells by redirecting recombinant lysosomal transporters to the cell surface. After surveying the current state of knowledge in this field, the review focuses on the sialic acid transporter sialin and shows how recent functional data using the above whole-cell approach shed new light on the pathogenesis of sialic acid storage disorders by revealing the existence of a residual transport activity associated with Salla disease. PMID- 18425436 TI - Molecular analysis of the GlcNac-1-phosphotransferase. AB - Modification of the carbohydrate chains of soluble lysosomal enzymes with mannose 6-phosphate residues is a prerequisite for their mannose 6-phosphate receptor dependent transport to lysosomes. GlcNac-1-phosphotransferase localized in the Golgi apparatus represents a hexameric alpha(2)beta(2)gamma(2) subunit complex and plays a key role in the formation of the mannose 6-phosphate recognition marker. Defects in the GlcNac-1-phosphotransferase complex cause two diseases, mucolipidosis type II and III, which are characterized by missorting and cellular loss of lysosomal enzymes, and lysosomal accumulation of storage material. The recent identification of two genes, GNPTAB and GNPTG, encoding the three subunits of GlcNac-1-phosphotransferase leads to an improvement of both pre- and postnatal diagnosis of affected individuals, and permits the analysis of structural requirements for efficient formation of mannose 6-phosphate residues on lysosomal enzymes. The alpha/beta subunits precursor matures by proteolytic cleavage and contains the catalytic activity as well as the capability to recognize lysosomal enzymes. The role of the gamma-subunits for activity, stability and oligomerization of the GlcNac-1-phosphotransferase subunits is still unclear. PMID- 18425437 TI - Serum prolactin as a tool for the follow-up of treated DHPR-deficient patients. AB - Deficiency of dihydropteridine reductase causes a variant form of phenylketonuria associated with a devastating neurological disease characterized by mental retardation, hypokinesis and other features relating to basal ganglia disorder. Hyperphenylalaninaemias with tetrahydrobiopterin deficiency make up about 1-3% of all hyperphenylalaninaemias. We describe three patients from Calabria, a southern region of Italy, who have a dihydropteridine reductase deficiency, caused by the same mutation (p.L14P) also found in the nearby region of Sicily. We report the evolution of clinical and biochemical data during the treatment of these patients where we used prolactin serum determination to adapt the specific therapy. This report suggests that serum prolactin levels can be a good biomarker for optimal dosage of hydroxylated precursors in long-term treatment monitoring. PMID- 18425439 TI - Lipid homeostasis in macrophages - implications for atherosclerosis. AB - In industrialized societies with excess food supply, obesity is an expanding problem. As a result of metabolic overload, besides obesity, insulin resistance, type-2 diabetes, dyslipidemia, hypertension, and atherosclerosis develop, which together make up the metabolic syndrome. The imbalance of lipid uptake, metabolism, and removal in many organs such as the liver, muscle, adipose tissue, vessel wall, and macrophages triggers organ transdifferentiation toward lipid storage phenotypes. Macrophages, foam cells, and osteoclasts in calcifying lesions are a hallmark of atherosclerosis and the metabolic syndrome, and must be regarded as an important therapeutic target. In this review, pathways regulating lipid homeostasis in macrophages are updated. These include lipid influx through different receptor entry pathways, the role of membrane microdomains, endolysosomal and cytosolic lipid storage leading to phospholipidosis, and lipid droplet accumulation or activation of lipid efflux either through the Golgi system or bypassing this organelle on the way to the plasma membrane. The interdependence of these pathways and pharmacological modifications are described. The monocyte innate immunity receptor complex in defining monocyte subpopulations and their role in cardiovascular disease is taken into account. The composition of certain molecular lipid species in membrane microdomains and other organelles is essential for cellular functions affecting raft dynamics, signal transduction, and membrane and organelle trafficking. It is very likely that the underlying defects in lipid-associated rare genetic diseases such as ABCA1 deficiency, Niemann-Pick disease type C, as well as the more frequent complex disorders associated with atherosclerosis and phospholipidosis are related to disturbances in membrane homeostasis, signal transduction, and cellular lipid metabolism. PMID- 18425438 TI - Surgical outcome of intravitreal bevacizumab and filtration surgery in neovascular glaucoma. AB - INTRODUCTION: The purpose of this study was to evaluate the surgical outcome of a trabeculectomy with mitomycin C (MMC) in neovascular glaucoma after an adjunctive treatment with intravitreal bevacizumab (Avastin(R); Genentech Inc, San Francisco, CA, USA) injection (IVB). METHODS: Six patients with NVG presented at the Department of Ophthalmology, Faculty of Medicine, Siriraj Hospital, Mahidol University, Thailand. After adequate panretinal photocoagulation (PRP) and maximal antiglaucoma medication therapy, these patients received IVB (1.25 mg in 0.05 ml) due to persistent neovascularisation of the iris (NVI). A fornix-based conjunctival flap trabeculectomy with MMC was performed within 4 weeks following administration of IVB. RESULTS: Three patients with proliferative diabetic retinopathy (PDR) and three patients with central retinal vein occlusion (CRVO) were enrolled in the study (mean age, 57 years). Absolute regression of NVI was observed within 1 week after IVB in four patients. In two patients NVI was reduced but still persisted. Mean intraocular pressure (IOP) decreased from 39.8 mmHg pre-operatively to 7.5 mmHg on the first postoperative day. No intra operative complications were noted. Two patients had postoperative hyphema, which resolved spontaneously within 1 week. During the mean follow up of 24.7 weeks, five patients had controlled IOP (range, 2-16 mmHg) without antiglaucoma medication. Two patients with PDR had improved visual acuity whereas two patients with CRVO lost pre-operative light perception. Recurrent NVI was subsequently detected in one patient who had uncontrolled IOP. This patient underwent transscleral diode laser cyclophotocoagulation and additional PRP. All patients were symptom-free at last visit. CONCLUSION: IVB is an effective modality to reduce intra-operative complications during trabeculectomy for neovascular glaucoma. The short-term outcomes following trabeculectomy with MMC are favourable. PMID- 18425440 TI - Autophagosome and phagosome. AB - Autophagy and phagocytosis are evolutionarily ancient processes functioning in capture and digestion of material found in the cellular interior and exterior, respectively. In their most primordial form, both processes are involved in cellular metabolism and feeding, supplying cells with externally obtained particulate nutrients or using portions of cell's own cytoplasm to generate essential nutrients and energy at times of starvation. Although autophagy and phagocytosis are commonly treated as completely separate biological phenomena, they are topologically similar and can be, at least morphologically, viewed as different manifestations of a spectrum of related processes. Autophagy is the process of sequestering portions of cellular interior (cytosol and intracellular organelles) into a membranous organelle (autophagosome), whereas phagocystosis is its topological equivalent engaged in sequestering cellular exterior. Both autophagosomes and phagosomes mature into acidified, degradative organelles, termed autolysosomes and phagolysosomes, respectively. The basic role of autophagy as a nutritional process, and that of phagocytosis where applicable, has survived in present-day organisms ranging from yeast to man. It has in addition evolved into a variety of specialized processes in metazoans, with a major role in cellular/cytoplasmic homeostasis. In humans, autophagy has been implicated in many health and disease states, including cancer, neurodegeneration, aging and immunity, while phagocytosis plays a role in immunity and tissue homeostasis. Autophagy and phagocytosis cooperate in the latter two processes. In this chapter, we briefly review the regulatory and execution stages of both autophagy and phagocytosis. PMID- 18425441 TI - Fine structure of the autophagosome. AB - This chapter describes the electron microscopic fine structure of early and late autophagic vacuoles in mammalian cells. Detailed instructions are given for the preparation of cells for conventional electron microscopy and for the identification of autophagic vacuoles by morphology. Electron microscopy remains one of the most accurate methods for quantitation of autophagic vacuole accumulation. Therefore, quantitation of autophagic vacuoles by electron microscopy and point counting is also described. Finally, a short description is given for preparation of ultra thin cryosections for immunogold labeling of autophagic vacuoles. PMID- 18425442 TI - Methods for assessing autophagy and autophagic cell death. AB - Autophagic (or type 2) cell death is characterized by the massive accumulation of autophagic vacuoles (autophagosomes) in the cytoplasm of cells that lack signs of apoptosis (type 1 cell death). Here we detail and critically assess a series of methods to promote and inhibit autophagy via pharmacological and genetic manipulations. We also review the techniques currently available to detect autophagy, including transmission electron microscopy, half-life assessments of long-lived proteins, detection of LC3 maturation/aggregation, fluorescence microscopy, and colocalization of mitochondrion- or endoplasmic reticulum specific markers with lysosomal proteins. Massive autophagic vacuolization may cause cellular stress and represent a frustrated attempt of adaptation. In this case, cell death occurs with (or in spite of) autophagy. When cell death occurs through autophagy, on the contrary, the inhibition of the autophagic process should prevent cellular demise. Accordingly, we describe a strategy for discriminating cell death with autophagy from cell death through autophagy. PMID- 18425443 TI - LC3 and Autophagy. AB - Microtubule-associated protein 1A/1B-light chain 3 (LC3) is a soluble protein with a molecular mass of approximately 17 kDa that is distributed ubiquitously in mammalian tissues and cultured cells. During autophagy, autophagosomes engulf cytoplasmic components, including cytosolic proteins and organelles. Concomitantly, a cytosolic form of LC3 (LC3-I) is conjugated to phosphatidylethanolamine to form LC3-phosphatidylethanolamine conjugate (LC3-II), which is recruited to autophagosomal membranes. Autophagosomes fuse with lysosomes to form autolysosomes, and intra-autophagosomal components are degraded by lysosomal hydrolases. At the same time, LC3-II in autolysosomal lumen is degraded. Thus, lysosomal turnover of the autophagosomal marker LC3-II reflects starvation-induced autophagic activity, and detecting LC3 by immunoblotting or immunofluorescence has become a reliable method for monitoring autophagy and autophagy-related processes, including autophagic cell death. Here we describe basic protocols to assay for endogenous LC3-II by immunoblotting, immunoprecipitation, and immunofluorescence. PMID- 18425444 TI - Amino acid regulation of autophagosome formation. AB - Amino acids are not only substrates for various metabolic pathways, but can also serve as signaling molecules controlling signal transduction pathways. One of these signaling pathways is mTOR-dependent and is activated by amino acids (leucine in particular) in synergy with insulin. Activation of this pathway inhibits autophagy. Because activation of mTOR-mediated signaling also stimulates protein synthesis, it appears that protein synthesis and autophagic protein degradation are reciprocally controlled by the same signaling pathway. Recent developments indicate that amino acid-stimulated mTOR-dependent signaling is subject to complex regulation. The mechanism by which amino acids stimulate mTORdependent signaling (and other signaling pathways), and its molecular connection with the autophagic machinery, is still unknown. PMID- 18425445 TI - Autophagic proteolysis of long-lived proteins in nonliver cells. AB - Autophagy is a cellular homeostasis pathway used to sustain cellular anabolic needs during times of nutrient or energy deprivation. Autophagosomes sequester cytoplasmic constituents, including macromolecules such as long-lived proteins. Upon fusion of autophagosomes with lysosomes, the engulfed cargo is degraded. The proteolysis of longlived proteins by macroautophagy is a standard, specific measure of autophagic degradation and represents an end-point assay for the pathway. The assay is based on a pulse-chase approach, whereby cellular proteins are radiolabeled by an isotopically marked amino acid, the short-lived, rapidly turned over, proteins are allowed to be degraded during a long chase period, and then the remaining, stable radiolabeled proteins are subjected to autophagic degradation. The classical application of this method has been in hepatocytes, but the recent growth of interest in autophagy has necessitated adaptation of this method in nonliver cells. Here we describe a protocol to quantify autophagic degradation of longlived proteins in macrophages. This chapter details the method of analyzing autophagic proteolysis in RAW264.7 mouse macrophages. PMID- 18425446 TI - Autophagosomes in GFP-LC3 Transgenic Mice. AB - Recent studies of the molecular mechanism of autophagy have made available several marker proteins for autophagosomes. These marker proteins allow us to identify autophagic structures easily and accurately by fluorescent microscopy. The most widely used marker for autophagosome is LC3, a mammalian homolog of Atg8. To analyze autophagy in whole animals, we generated GFP-LC3 transgenic mice and describe here how we determine the occurrence of autophagy in vivo using this mouse model. PMID- 18425447 TI - Experimental control and characterization of autophagy in Drosophila. AB - Insects such as the fruit fly Drosophila melanogaster, which fundamentally reorganize their body plan during metamorphosis, make extensive use of autophagy for their normal development and physiology. In the fruit fly, the hepatic/adipose organ known as the fat body accumulates nutrient stores during the larval feeding stage. Upon entering metamorphosis, as well as in response to starvation, these nutrients are mobilized through a massive induction of autophagy, providing support to other tissues and organs during periods of nutrient deprivation. High levels of autophagy are also observed in larval tissues destined for elimination, such as the salivary glands and larval gut. Drosophila is emerging as an important system for studying the functions and regulation of autophagy in an in vivo setting. In this chapter we describe reagents and methods for monitoring autophagy in Drosophila, focusing on the larval fat body. We also describe methods for experimentally activating and inhibiting autophagy in this system and discuss the potential for genetic analysis in Drosophila to identify novel genes involved in autophagy. PMID- 18425448 TI - Analysis of autophagosome membrane cycling by fluorescence microscopy. AB - Autophagy is a physiological process functionally linked to cellular dynamics during starvation, cardiomyopathies, neurodegeneration, cellular immunity, and certain cancers. Although nearly 30 autophagy-related (ATG) genes have been identified and characterized, the molecular mechanisms of this process are only partially understood. One aspect of the pathway that has been intensely studied is the identity of the membrane source for newly formed autophagosomes. Although it occurs at a basal level, autophagy is an inducible process. The process of autophagosome formation involves recruitment and delivery of membrane and recycling of Atg proteins. Despite continuing attempts to identify the source of the autophagosome membrane, we are only recently beginning to understand the nature of autophagosome formation and the role of membrane protein cycling in this process. There now exists an assay utilizing fluorescence microscopy to monitor the localization, and therefore the movement, of membrane-associated Atg proteins. We describe here a method that allows visualization of Atg membrane proteins in order to observe their potential source membranes and also to determine the temporal order of action of other Atg proteins with regard to their movement. PMID- 18425449 TI - Protein trafficking into autophagosomes. AB - The methods described are designed to enable the assignment of an intracellular localization of secretory proteins, either soluble or membrane associated, to later secretory compartments, such as the trans-Golgi network (TGN) or endosome. These two subcellular compartments are closely linked through extensive protein trafficking, in both an anterograde and a retrograde direction. These compartments are likely to be important in the formation of autophagosomes during the process of autophagy. Our current knowledge of how autophagosomes form is scarce, and further investigation into the role that other subcellular compartments have in this process is needed. PMID- 18425450 TI - Sphingolipids in macroautophagy. AB - Sphingolipids are constituents of biological membranes. Ceramide and sphingosine 1-phosphate (S1P) also act as second messengers and are part of a rheostat system, in which ceramide promotes cell death and growth arrest, and S1P induces proliferation and maintains cell survival. As macroautophagy is a lysosomal catabolic mechanism involved in determining the duration of the lifetime of cells, we raised the question of its regulation by sphingolipid messengers. Using chemical and genetic methods, we have shown by GFP-LC3 staining and analysis of the degradation of long-lived proteins that both ceramide and S1P stimulate autophagy. PMID- 18425451 TI - Molecular links between autophagy and apoptosis. AB - Macroautophagy (herein referred to as autophagy) contributes to the control of life and death throughout the animal and plant kingdoms. Bilateral links have been found between apoptosis and autophagy where inducers of apoptosis also induce autophagy and vice versa. In some cases, autophagy delays the onset of apoptosis and thus prolongs life although it may also promote apoptosis and other forms of cell death. It is thus of great biological and medical interest to understand the molecular connections between these two pathways, and try to utilize-or block-them selectively to aid induction of cell death (e.g., cancer cells) or inhibit death (e.g., in degenerative disorders). This chapter describes methods for studying apoptotic induction of autophagy and its effects on cell function. We also discuss potential pitfalls. Although cell lines are used as model systems, the substances and methods described here can be applied to primary cells and tissues. PMID- 18425452 TI - Clearance of mutant aggregate-prone proteins by autophagy. AB - The accumulation of mutant aggregate-prone proteins is a feature of several human disorders, collectively referred to as protein conformation disorders or proteinopathies. We have shown that autophagy, a cytosolic, non-specific bulk degradation system, is an important clearance route for many cytosolic toxic, aggregate-prone proteins, like mutant huntingtin and mutant alpha-synucleins. Induction of autophagy enhances the clearance of both soluble and aggregated forms of the mutant protein, and protects against toxicity caused by these mutations in cell, fly, and mouse models. Inhibition of autophagy has opposite effects. Thus, the autophagic pathway may represent a possible therapeutic target in the treatment of certain protein conformation disorders. PMID- 18425453 TI - Localization and MHC class II presentation of antigens targeted for macroautophagy. AB - Intracellular antigens can be presented on major histocompatibility complex (MHC) class II molecules after degradation via macroautophagy. To enhance MHC class II presentation of potential vaccine antigens, we have developed a method to target antigens for autophagic degradation via fusion to the Atg8/LC3 protein: Atg8/LC3 is specifically incorporated into autophagosomes via coupling to phosphatidylethanolamine, and subsequently degraded in MHC class II loading compartments (MIICs). Antigens fused to the N-terminus of Atg8/LC3 follow the same pathway and get preferentially presented on MHC class II molecules. The localization of Atg8/LC3 fusion antigens in MIICs can be visualized by confocal microscopy, and MHC class II presentation can be quantified in a presentation assay with antigen-specific CD4(+) T-cell clones. These assays are good measures of autophagosome formation and lysosomal degradation of macroautophagy cargo and therefore are useful for studying regulation of the autophagic pathway under various experimental conditions and physiological perturbations. PMID- 18425454 TI - Chaperone-mediated autophagy. AB - Chaperone-mediated autophagy (CMA) is the only type of autophagy in mammalian cells able to selectively degrade cytosolic proteins in lysosomes. CMA is maximally activated in response to stressors such as prolonged starvation, exposure to toxic compounds, or oxidative stress. We have found that CMA activity decreases in aging and in some age-related disorders such as Parkinson's disease. Impaired CMA under these conditions may be responsible for the accumulation of damaged proteins inside cells and for their higher vulnerability to stressors. In contrast to other forms of autophagy, where substrates are engulfed or sequestered along with other cytosolic components, CMA substrates are translocated one-by-one across the lysosomal membrane. Changes in the levels/activity of the lysosomal components required for substrate translocation can be used to stimulate CMA activity. However, the most unequivocal method to measure CMA is by directly tracking the translocation of substrate proteins into isolated lysosomes. PMID- 18425455 TI - Microautophagy in the yeast Saccharomyces cerevisiae. AB - Microautophagy involves direct invagination and fission of the vacuolar/lysosomal membrane under nutrient limitation. In Saccharomyces cerevisiae microautophagic uptake of soluble cytosolic proteins occurs via an autophagic tube, a highly specialized vacuolar membrane invagination. At the tip of an autophagic tube vesicles (autophagic bodies) pinch off into thevacuolar lumen for degradation. Formation of autophagic tubes is topologically equivalent to other budding processes directed away from the cytosolic environment, e.g., the invagination of multivesicular endosomes, retroviral budding, piecemeal microautophagy of the nucleus and micropexophagy. This clearly distinguishes microautophagy from other membrane fission events following budding toward the cytosol. Such processes are implicated in transport between organelles like the plasma membrane, the endoplasmic reticulum (ER), and the Golgi. Over many years microautophagy only could be characterized microscopically. Recent studies provided the possibility to study the process in vitro and have identified the first molecules that are involved in microautophagy. PMID- 18425456 TI - EM analysis of phagosomes. AB - Electron microscopy (EM) is the only technique that can combine sensitive protein detection methods with detailed information on the substructure of cellular compartments. The purpose of this chapter is to describe some of the methods at the EM level that can be used to analyze the spatial organization of cell organelles with respect to phagosomes or vacuoles in which pathogens are sequestered, characterize the compartment in which pathogens are harbored, ie immature phagosomes, phagolysosomes, autophagosomes, and ER-derived vacuoles, to cite a few, and decipher the molecular mechanisms involved in survival of pathogens within infected host cells. PMID- 18425457 TI - Analysis of phosphoinositide dynamics during phagocytosis using genetically encoded fluorescent biosensors. AB - Phosphoinositide signaling is essential for successful phagocytosis. Phosphoinositides regulate processes such as actin assembly and the recruitment of molecular motors required for ingestion, as well as fusion events required for the maturation of the phagosome. Phosphoinositides not only serve as substrates for the generation of second messengers, but also function to anchor to the membrane cytosolic proteins that contain phosphoinositide-binding motifs. Conventional methods for the detection of phosphoinositides involve their extraction from the cells and separation by chromatographic procedures. These approaches are laborious and expensive and fail to provide spatio-temporal information, which is critical when analyzing localized and transient phenomena like phagocytosis. In this chapter we describe a method to monitor phosphoinositides dynamically by transfection of fluorescently tagged probes (biosensors) into cultured macrophages. These biosensors are based on the fusion of phosphoinositide-binding protein domains with fluorescent proteins. Some specifications for live cell imaging of such phosphoinositide-specific probes are also provided. PMID- 18425458 TI - In vitro phagosome-endosome fusion. AB - Phagolysosome biogenesis plays a pivotal role in elimination of foreign particles and pathogens by leukocytes. This process is achieved by multiple cycles of membrane fusion between the phagosome and the endosomal system. In vitro reconstitution of phagosome fusion with endosomes is instrumental in studying this intricate process. Such an assay is also invaluable for the identification of effectors produced by intracellular pathogens that may hamper the pathway. In this chapter we describe a highly sensitive and relatively rapid method to measure fusion between phagosomes and early, as well as late, endosomal compartments. The assay is based on the formation of a stable biotin-streptavidin complex upon fusion between biotinylated-peroxidase loaded endosomes and magnetic streptavidin conjugated bead-containing phagosomes. Fusion is quantified using a fluorescence-based detection method that measures the peroxidase activity associated with the beads. PMID- 18425459 TI - Real-time spectrofluorometric assays for the lumenal environment of the maturing phagosome. AB - The ultimate goal of phagosomal maturation is the delivery of internalized, particulate cargo to acidic, hydrolytically competent compartments capable of mediating its degradation. Here we outline in detail three fluorometric techniques that allow the study of phagosomal maturation in macrophages by quantifying functionally important features of the lumenal environment of the developing phagosome in real time. The first assay utilizes a particle restricted, pH-sensitive fluorochrome to measure the acidification of the phagosome. The second reports on the development of the proteolytic capacity of the phagosome by following the hydrolysis of a fluorogenic, generic proteinase substrate. The third quantifies the accumulation of lysosomal constituents within the phagosome by measuring the fluorescence resonance energy transfer (FRET) efficiency between a particle-restricted, donor fluor and a fluid phase acceptor fluor that had been chased previously into lysosomes. The assays are described as population-based methodologies utilizing a spectrofluorometer but, alternatively, can be adapted readily to confocal-based technologies for single phagosomal measurements. PMID- 18425460 TI - Monitoring time-dependent maturation changes in purified phagosomes from Dictyostelium discoideum. AB - The amoeba Dictyostelium discoideum is an established model to study phagocytosis. The sequence of events leading to the internalization and degradation of a particle is conserved in D. discoideum compared to metazoan cells. As its small haploid genome has been sequenced, it is now amenable to genome-wide analysis including organelle proteomics. Therefore, we adapted to Dictyostelium the classical protocol to purify phagosomes formed by ingestion of latex beads particles. The pulse-chase protocol detailed here gives easy access to pure, intact, and synchronized phagosomes from representative stages of the entire process of phagosome maturation. Recently, this protocol was used to generate individual temporal profiles of proteins and lipids during phagosome maturation generating a proteomic fingerprint of six maturation stages (1). In addition, immunolabeling of phagosomes on a coverslip was developed to visualize and quantitate antigen distribution at the level of individual phagosomes. PMID- 18425461 TI - Large-scale purification of latex bead phagosomes from mouse macrophage cell lines and subsequent preparation for high-throughput quantitative proteomics. AB - Phagocytosis involves the engagement of a diverse array of cell surface receptors whose signal must be integrated on the membrane of the forming phagosomal cup. This method enables the quantitative proteomic analysis of phagosome fractions derived from phagocytes stimulated under two different conditions, thus allowing the complexity of phagosomal signaling to be analyzed in terms of the quantitative changes in phagosomal fraction protein content. PMID- 18425462 TI - Class II MHC antigen processing in phagosomes. AB - Phagocytic antigen-presenting cells (APCs) are involved in innate and adaptive immune responses to bacteria. Adaptive responses to bacteria involve processing of bacterial antigens for presentation by class II major histocompatibility complex (MHC II) molecules and class I MHC (MHC I) molecules to stimulate CD4(+) and CD8(+) T cells, respectively. To examine the role of phagosomes in processing of antigens for presentation by MHC II molecules to CD4(+) T cells, phagosomes have been biochemically and functionally analyzed by a variety of techniques that include flow analysis (flow organellometry), SDS-PAGE/Western blotting, and an antigen-presenting organelle assay. Using these techniques, we have demonstrated that phagosomes containing latex beads or Mycobacterium tuberculosis (MTB) contain components of the MHC II processing pathway and support the formation of peptide-MHC II complexes. PMID- 18425463 TI - Analyzing association of the endoplasmic reticulum with the legionella pneumophila-containing vacuoles by fluorescence microscopy. AB - A unique feature of the intracellular life cycle of Legionella pneumophila is the interaction between the vacuole in which L. pneumophila resides and the endoplasmic reticulum of the host cell. This interaction is crucial for L. pneumophila to establish a niche in which the bacteria can replicate intracellularly. Microscopic analysis of endoplasmic reticulum (ER) markers during infection yields information regarding the nature of the recruited vesicles as well as the kinetics of their recruitment. The recruitment of YFP KDEL, GFP-p58, calnexin, and myc-Sec22b to the L. pneumophila - containing vacuole can be assessed by fluorescence microscopy. Methods for detection of these various ER markers during infection of mammalian cells by L. pneumophila are described. PMID- 18425464 TI - Fractionation of the Coxiella burnetii parasitophorous vacuole. AB - Coxiella burnetii is a bacterial obligate intracellular pathogen that replicates within a spacious parasitophorous vacuole (PV) with lysosomal characteristics. The pathogen actively participates in the biogenesis of its PV by synthesizing proteins that mediate vesicular interactions. Both C. burnetii and host factors that regulate PV formation are likely localized to the PV membrane, and their identification would be aided by an efficient method for isolating the C. burnetii vacuole. To this end, we developed a method to separate intact PV from host cell material that relies on fusion of the vacuole with latex bead containing phagosomes (LBP). Sequestration of latex beads by the C. burnetii PV increases the vacuole's buoyant density and facilitates its fractionation on a sucrose step gradient. Transmission electron microscopy confirms the isolation of intact PV-containing latex beads from infected MH-S murine alveolar macrophage like cells. Immunoblotting demonstrates that C. burnetii PV lysates are dramatically enriched for the late endosome/lysosome markers LAMP-1 and LAMP-2 when compared to total host cell lysates. Conversely, PV preparations are devoid of p62 and GM130, markers of the nucleus and Golgi apparatus, respectively, indicating effective separation of the vacuole from these host cell compartments. Two-dimensional gel electrophoresis and immunoblotting reveal distinct protein differences between C. burnetii PV and LBP. Identification of proteins unique to the PV membrane will yield important insight into C. burnetii-host interactions. PMID- 18425465 TI - Bacterial phagosome acidification within IFN-gamma-activated macrophages: role of host p47 immunity-related GTPases IRGs). AB - Interferon-gamma IFN-gamma)-induced remodeling of the bacterial phagosome for pathogen clearance elicits the aid of a new family of GTPases termed the p47 IRGs. Members of this group reside primarily on ER-Golgi membranes before translocating to the nascent phagosome within minutes of bacterial uptake. Recruitment of p47 IRGs coincides with the acquisition of phagosome maturation and autophagy markers as well as enhanced acidification of this organelle. Here we describe a simple spectrofluorometric assay to measure luminal acidification of the bacterial phagosome within intact cells such as macrophages. This method can be applied to study the phagosomal pH pH_pg) of activated cells infected with a variety of infectious microorganisms and the roles played by members of the p47 IRG family in auto)phagolysosome biogenesis. PMID- 18425466 TI - SopE-mediated recruitment of host Rab5 on phagosomes inhibits Salmonella transport to lysosomes. AB - Phagocytosis is a process by which invading organisms are taken up by macrophages and targeted to the lysosomes, where they are degraded. However, many pathogens modulate this central process of macrophage-mediated killing by inhibiting their transport to the lysosomes through a variety of pathogen-derived mechanisms. Given the importance of Rab proteins in the regulation of intracellular transport pathways, we investigated the role of different host endocytic Rabs on the maturation of Salmonella-containing phagosomes in macrophages. Initially, we have developed a ligand mixing assay to measure the transport of the Salmonella containing phagosomes to lysosomes. Using this assay we have shown that Salmonella decline their transport to the lysosomes. In order to determine whether inhibition of Salmonella transport to lysosomes is due to their sustained fusion with early endosomes, we have developed an in vitro fusion assay between Salmonella-containing phagosomes and early endosomes. Here, we have discussed how these methodologies are helpful to determine the mechanism of evasion of Salmonella transport to the lysosomes. PMID- 18425467 TI - The Mycobacterium tuberculosis phagosome. AB - Tuberculosis is currently the most devastating human bacterial disease, causing millions of deaths annually and infecting an overwhelming percentage of the global population. Its success as a scourge lies in the ability of Mycobacterium tuberculosis to prevent normal phagolysosome biogenesis, essential to the destruction of invading microorganisms, inside macrophages. Recent work has identified host GTPases involved in the block of normal phagolysosome biogenesis during mycobacterial infection and has provided a set of methods, in particular efficient macrophage transfection, which will prove essential in examining the role of host effectors in this process. PMID- 18425468 TI - Spectral karyotyping (SKY): applications in prenatal diagnostics. AB - The method of spectral karyotyping (SKY) is based on a combination of the technologies of charge-coupled device imaging and spectrometry. The engineering feasibility has been realized in the SpectraCube system from Applied Spectral Imaging Inc., and it allows the simultaneous identification of all 24 human chromosomes. This is performed by characterizing the spectral signature of every image pixel in relation to a fluorochrome combinatorial library translating the image and spectral information into chromosome classification. Applications for SKY include pre- and postnatal characterization of certain numerical and structural rearrangements and complex karyotypes and highly informative analysis of sample materials with only single or few cells available for investigation. PMID- 18425469 TI - Characterization of prenatally assessed de novo small supernumerary marker chromosomes by molecular cytogenetics. AB - Small supernumerary marker chromosomes (sSMC) are structurally abnormal chromosomes that cannot be identified or characterized unambiguously by conventional banding cytogenetics alone, and they are generally equal in size or smaller than a chromosome 20 of the same metaphase spread. sSMC are reported in 0.043% of newborn infants and 0.075% of prenatal cases. Molecular cytogenetics is necessary to characterize the origin of an sSMC, and many highly sophisticated approaches are available throughout the literature for their comprehensive description. However, because in a prenatal diagnostic laboratory such techniques are not available, I suggest here a straightforward scheme to characterize at least the sSMC's chromosomal origin as quickly as possible. Based on this scheme, it is possible to compare the actual present case with similar cases from the literature, which are summarized on http://www.med.uni jena.de/fish/sSMC/00START.htm./ For a more wide-ranging sSMC characterization, a specialized laboratory should be contacted, e.g., my laboratory. PMID- 18425470 TI - Rapid prenatal aneuploidy screening by fluorescence in situ hybridization (FISH). AB - The most common aneuploidies in prenatal diagnostics of the second trimenon are trisomies of chromosomes 13, 18, and 21 and gonosomal abnormalities. To detect these trisomies as quickly as possible after amniocentesis, besides using polymerase chain reaction, fluorescence in situ hybridization (FISH), applying corresponding centromeric or locus-specific probes, is the method of choice. Results of a rapid prenatal aneuploidy screening in uncultured amniocytes by using FISH are available within 24 hr or less. However, care has to be taken against possible pitfalls in connection with the commercially available probe sets and thus interpretation of results in general. Here, we explain how rapid prenatal aneuploidy screening is performed using the Food and Drug Administration approved Aneu Vysion kit (Abbott/Vysis), and a review is given of drawbacks and opportunities of this method. PMID- 18425472 TI - Prenatal diagnosis using array CGH. AB - Microarray-based comparative genomic hybridization (array CGH) is a fast and high resolution approach to the diagnosis of a large number of genetic syndromes associated with loss or gain of the human genome. This technology has proven to be useful in several clinical settings, including postnatal diagnosis of mental retardation, developmental delay, and congenital malformation syndromes. We describe the use of array CGH for prenatal diagnosis of a range of chromosomal syndromes associated with congenital malformations visible by ultrasound. The procedure is reproducible in a clinical setting and provides reliable results in a short period (approximately 5 days). Thus, depending on the array used, array CGH may develop into an excellent tool for prenatal diagnosis. PMID- 18425471 TI - Application of multi-PRINS to simultaneously identify chromosomes 18, X, and Y in prenatal diagnosis. AB - Since its discovery by Koch in 1989, primed in situ labeling (PRINS) reaction provides an alternative approach for direct detection of human chromosomes. The multiple color (multi)-PRINS technique can simultaneously and specifically display different chromosomes with different colors in the same metaphase or interphase nucleus by using sequential labeling of different chromosome targets. We developed a triple-PRINS reaction on uncultured amniotic cells by omitting the blocking step and taking advantage of mixing two fluorochromes (fluorescein and rhodamine) to create a third color for simultaneous detection in the same amniocytes of three different chromosome targets, e.g., chromosomes 18, X, and Y. Fluorescent signals corresponding to chromosomes 18, X, and Y were shown as yellow, red, and green color spots, respectively. Multi-PRINS is as accurate and reliable as multicolor fluorescent in situ hybridization (multi-FISH) for the detection of aneuploidies involving chromosomes 18, X, and Y. Furthermore, multi PRINS represents a faster and more cost-effective alternative to FISH for prenatal testing of aneuploidy in uncultured amniocytes. PMID- 18425473 TI - Prenatal detection of chromosome aneuploidy by quantitative fluorescence PCR. AB - Autosomal chromosome aneuploid pregnancies that survive to term, namely, trisomies 13, 18, and 21, account for 89% of chromosome abnormalities with a severe phenotype. They are normally detected by full karyotype analysis of cultured cells. The average reporting time for a prenatal karyotype analysis is approximately 14 days, and in recent years, there has been increasing demand for more rapid prenatal results with respect to the common chromosome aneuploidies, to relieve maternal anxiety and facilitate options in pregnancy. The rapid tests that have been developed negate the requirement for cultured cells, instead directly testing cells from the amniotic fluid or chorionic villus sample, with the aim of generating results within 48 h of sample receipt. Interphase fluorescence in situ hybridization is the method of choice in some genetic laboratories, usually because the expertise and equipment are readily available. However, a quantitative fluorescence (QF)-PCR-based approach is more suited to a high-throughput diagnostic service. This approach has been investigated in a small number of pilot studies and reported as a clinical diagnostic service in many studies. It may be used as a stand-alone test or as an adjunct test to full karyotype analysis, which subsequently confirms the rapid result and scans for other chromosome abnormalities not detected by the QF-PCR assay. PMID- 18425474 TI - Real-time quantitative PCR for the detection of fetal aneuploidies. AB - In prenatal analysis, one of the major concerns is the detection of fetal aneuploidies. Several molecular methods have been described recently for the rapid analysis of amniotic fluid and chorionic villi. Fluorescence in situ hybridization (FISH) and polymerase chain reaction (PCR) of short tandem repeats are already implemented in prenatal laboratories and permit the evaluation of the major chromosomal aberrations within 24 h. However, both methods have their disadvantages. The advent of real-time PCR has revolutionized the measurement of nucleic acid copy numbers in recent years. Quantitative PCR, a term that was only 10 years ago considered to be an oxymoron, is now widely accepted. We demonstrated previously the feasibility of detection of trisomy 21 by real-time PCR, and here we describe the modified test that permits simultaneous analysis of trisomies 18 and 21. This approach has been demonstrated in a recent large-scale analysis, and it is presented in detail in this chapter. PMID- 18425475 TI - MLPA for prenatal diagnosis of commonly occurring aneuploidies. AB - Multiplex ligation-dependent probe amplification (MLPA) is a new method to determine the copy number of up to 45 genomic DNA sequences in a single multiplex polymerase chain reaction (PCR)-based reaction. In contrast to standard multiplex PCR, only one pair of PCR primers is used. MLPA reactions with currently commercial available kits result in very reproducible gel patterns with fragments of 130 to 480 bp that can be analyzed by sequence type electrophoresis. Comparison of this gel pattern to that obtained from a control sample indicates which sequences show an aberrant copy number. PMID- 18425476 TI - Maldi-TOF mass spectrometry for trisomy detection. AB - Matrix-associated laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry is a tool currently under investigation for use in prenatal detection of abnormalities in chromosome number, such as trisomy 21. Because of its ability to detect extremely small differences in mass, even to the level of a single nucleotide difference, this method can be applied to the detection of single-nucleotide polymorphisms (SNPs), which when present in a heterozygous state, can yield quantitative information regarding chromosome status from diagnostic specimens such as amniotic fluid or chorionic villus samples. MALDI TOF mass spectrometry has several potential advantages over traditional karyotyping methods, including its amenability to high-throughput analyses and its nonreliance on prior cell culture. The method described here is based on the MassEXTEND protocol developed by Sequenom, Inc., although any mass spectrometry platform sensitive enough to detect the small difference in mass between SNPs could be applied for this purpose. PMID- 18425477 TI - Rapid detection of fetal Mendelian disorders: thalassemia and sickle cell syndromes. AB - The inherited disorders of hemoglobin synthesis constitute the most common monogenic diseases worldwide. The clinical severity of beta-thalassemia major and the sickle cell syndromes targets them as priority genetic diseases for prevention programs, which incorporates population screening to identify heterozygotes,with the option of prenatal diagnosis for carrier couples. Rapid genotype characterization is fundamental in the diagnostic laboratory, especially when offering prenatal diagnosis. The application of real-time polymerase chain reaction (PCR) provides a means for rapid and potentially high-throughput assays, without compromising accuracy. It has several advantages over endpoint PCR analysis, including the elimination of post-PCR processing steps and a wide dynamic range of detection with a high degree of sensitivity. Although there are >180 mutations associated with the beta- thalassemia and sickle cell syndromes, the relatively small size of the beta-globin gene (<2,000 base pairs) and the proximity of most mutations facilitates the design of a minimal number of real time PCR assays by using the LightCycler system (Roche Diagnostics [Hellas] A.E., Athens, Greece), which are capable of detecting the majority of most common beta gene mutations worldwide. These assays are highly appropriate for rapid genotyping of parental and fetal DNA samples with respect to beta-thalassemia and sickle cell syndromes. PMID- 18425478 TI - Rapid detection of fetal Mendelian disorders: Tay-Sachs disease. AB - Tay-Sachs disease is an autosomal recessive storage disease caused by the impaired activity of the lysosomal enzyme hexosaminidase A. In this fatal disease, the sphingolipid GM2 ganglioside accumulates in the neurons. Due to high carrier rates and the severity of the disease, population screening and prenatal diagnosis of Tay-Sachs disease are routinely carried out in Israel. Laboratory diagnosis of Tay-Sachs is carried out with biochemical and DNA-based methods in peripheral and umbilical cord blood, amniotic fluid, and chorionic villi samples. The assay of hexosaminidase A (Hex A) activity is carried out with synthetic substrates, 4-methylumbelliferyl-6-sulfo-N-acetyl-beta-glucosaminide (4-MUGS) and 4-methylumbelliferil-N-acetyl-beta-glucosamine (4-MUG), and the DNA-based analysis involves testing for the presence of specific known mutations in the alpha-subunit gene of Hex A. Prenatal diagnosis of Tay-Sachs disease is accomplished within 24-48 h from sampling. The preferred strategy is to simultaneously carry out enzymatic analysis in the amniotic fluid supernatant or in chorionic villi and molecular DNA-based testing in an amniotic fluid cell pellet or in chorionic villi. PMID- 18425479 TI - Arrayed primer extension reaction for genotyping on oligonucleotide microarray. AB - Arrayed primer extension reaction (APEX) is a straightforward and robust enzymatic genotyping method in which hundreds to thousands of variations in the genome are simultaneously analyzed in a single multiplexed reaction. It differs from allele-specific hybridization in that the genotype information in APEX is obtained by single base extension, performed by a specific DNA polymerase, together with four different dye terminators. This approach ensures highly specific discrimination without allele-specific hybridization, because the primer to be extended anneals just adjacent to the DNA base that needs to be identified. Selection of primers for specific sites or their consecutive placement in tiled format, shifting them by one base, permits single-nucleotide polymorphism analysis, mutation detection, or resequencing of the DNA template. Careful primer design also permits the analysis of insertions, deletions, splice variants, gene copy numbers, or CpG islands within the genome for gene methylation studies, by performing additional bisulfite reactions. PMID- 18425480 TI - A fast microelectronic array for screening and prenatal diagnosis of beta thalassemia. AB - The electronic microchip is a recently developed technology for the fast and reliable detection of known single-nucleotide polymorphisms (SNPs) in the genome. The DNA fragment to be analyzed is directed electrophoretically into the chip, and then it is hybridized with fluorescent-tagged DNA probes specific for the mutant and wild-type sequences. The presence or absence of the mutation is detected by the fluorescence signal. Electronic stringency provides quality control for the hybridization process and ensures that any bound pairs of DNA are truly complementary; the microchip can be easily customized by the end user, allowing for assembly of specific probes onto the microchip to perform individualized analyses. Assays for 10 frequent mutations in the beta-globin gene causing beta-thalassemia and sickle cell anemia are presented that can be applied, in turn, to population screening or family study and prenatal diagnosis in single cases. PMID- 18425481 TI - RHD genotyping from maternal plasma: guidelines and technical challenges. AB - Rhesus D (RhD) blood group incompatibility between mother and fetus can occasionally result in maternal alloimmunization where the resultant anti-D can cross the placenta and attack the fetal red cells, which in worse case scenarios can cause fetal anemia and ultimately death. Fetal RHD genotyping was introduced in the mid-1990s after the molecular characterization of the RH genes as an aid to the clinical management of these cases. Initially, these tests used fetal DNA extracted invasively from chorionic villus and amniocyte samples. RHD genotyping of fetuses carried by RhD-negative women has become the first large-scale application of noninvasive prenatal diagnosis (NIPD). Initially the real-time polymerase chain reaction (PCR)-based tests were devised to characterize free fetal DNA in maternal plasma and serum, and RHD genotyping was a convenient assay to develop this exciting new technology, because the accuracy of tests could easily be confirmed after the simple RhD phenotyping of fetal cord blood cells after birth. "First generation" RHD genotyping tests were based on the incorrect concept that all D-negative phenotypes were caused by a complete RHD gene deletion. Thus, it was a relatively simple task to develop diagnostic PCR strategies based on the detection of RHD where D-negative genomes will completely lack RHD. Subsequent research into the molecular basis of D-negative phenotypes revealed that a significant number of D-negative genomes possess fragments of, or mutated RHD genes, the most notable of which is the RHD pseudogene found in Africans. Thus, more comprehensive RHD genotyping tests have evolved to differentiate these alleles, and are more appropriate in the diagnosis of multi ethnic population groups such as those found in Europe and North America. Many European Union countries have suggested the mass application of RHD NIPD for all fetuses carried by D-negative women. This is of clear benefit, because most RhD prophylaxis programs have switched to antenatal administration. This will help conserve anti-D stocks and it will prevent unnecessary administration of a human derived blood product to a vulnerable patient group. Although anti-D stocks are inherently safe, there is a moral obligation to eliminate unnecessary administration of it because there have been instances of hepatitis C infection due to contamination. Furthermore, as yet undescribed viruses may be contaminants of blood products. Mass-scale RHD NIPD will shortly be implemented in several countries in the European Community as a consequence. PMID- 18425482 TI - Isolation of cell-free DNA from maternal plasma using manual and automated systems. AB - Cell-free fetal DNA present in the maternal circulation holds great potential for noninvasive prenatal diagnosis and analysis of fetal genetic traits. However, only approximately 3-6% of total DNA in the maternal plasma is of fetal origin. Because of its scarcity in the maternal circulation, various methods have been developed and tested to optimize the extraction of this rare material from plasma. Here, we first describe the commonly used protocol for separating plasma from whole blood samples. We also describe two commercially available methods for the extraction of cell-free DNA from maternal plasma, which we have found particularly straightforward and easy to use: a manual method using the High Pure PCR Template Preparation kit (Roche Diagnostics) and an automated system using the MagNA Pure LC instrument (Roche Diagnostics). Use of the methods described here will help to ensure maximum yield and purity of cell-free fetal DNA extracted from maternal plasma samples for downstream analyses. PMID- 18425483 TI - Fetal DNA: strategies for optimal recovery. AB - For fetal DNA extraction, in principle each DNA extraction method can be used; however, because most methods have been optimized for genomic DNA from leucocytes, we describe here the methods that have been optimized for the extraction of fetal DNA from maternal plasma and validated for this purpose in our laboratories. The use of the QIAamp DSP Virus kit (QIAGEN), the QIAamp DNA Blood Mini kit (QIAGEN), and the Magna Pure LC (Roche) is based on the kit components provided by the respective companies. However, we noticed that the yield of fetal DNA from maternal plasma can be increased when higher volumes are processed or some slight modifications of the protocols provided by the manufacturer are followed. Here, we also describe an in-house method that allows the specific capture of target molecules in an extremely low volume by using magnetic beads and magnetic tips. This method can be either performed by hand, or it can be adapted to a commercially available pipetting workstation. PMID- 18425484 TI - Quantification of circulatory fetal DNA in the plasma of pregnant women. AB - The analysis of cell-free fetal DNA in the circulation of the pregnant woman plays the pivotal role in noninvasive prenatal research. Here, we describe an improved method for the quantification of male DNA, which is a valuable research tool for the quantification of fetal DNA. The quantification of fetal DNA serves two main purposes. First, the levels may indicate certain pregnancy-related disorders such as preeclampsia even before onset of the disease; thus, the quantification may serve as a marker for early detection. Second, extraction and enrichment strategies of the fetal DNA compartment are important factors in the development and implementation of clinical tests, such as detection of fetal sex, Rhesus D status, point mutations, and aneuploidies. In this context, the quantification of fetal DNA is an important tool for the evaluation of protocols. PMID- 18425485 TI - Detection and quantification of fetal DNA in maternal plasma by using LightCycler technology. AB - Since the demonstration of cell-free fetal DNA in maternal circulation, its aberrational quantification has been explored and demonstrated in numerous clinical situations. Moreover, several centers have begun to use detection and quantification of fetal DNA to diagnose fetal genetic status with high reliability, such as fetal RhD. One of the methods for analyzing cell-free fetal DNA is quantification by using LightCycler technology. With the advent of various fluorescent reporters and greater stability of kit's reagent, real-time quantitative polymerase chain reaction by using LightCycler technology is relatively simple and fast. PMID- 18425486 TI - Size fractionation of cell-free DNA in maternal plasma and its application in noninvasive detection of fetal single gene point mutations. AB - Recent studies have shown that cell-free fetal DNA in maternal plasma can be enriched by means of size fractionation. This technique makes use of the smaller size of fetal DNA fragments compared with maternal DNA fragments isolated simultaneously. On this basis, a highly improved detection of fetal single gene point mutations is permitted. Here, we introduce the use of agarose gel electrophoresis for the size fractionation of cell-free DNA from maternal plasma and the detection of fetal beta-thalassemia mutations in the size-fractionated cell-free DNA by using peptide nucleic acid-clamping polymerase chain reaction (PCR) combined with an allele-specific real-time PCR assay. Matrix-assisted laser desorption ionization/time of flight mass spectrometry has been reliably used for detection of fetal single gene point mutations in maternal plasma. We also present its use for genotyping paternally inherited single-nucleotide polymorphism alleles in the size-fractionated cell-free DNA from maternal plasma. PMID- 18425487 TI - Maldi-TOF mass spectrometry for analyzing cell-free fetal DNA in maternal plasma. AB - The discovery of cell-free fetal DNA in the plasma and serum of pregnant women has opened a new window for noninvasive prenatal diagnosis. Robust detection and quantification have been achieved when the fetal DNA sequence of interest does not have a maternal counterpart (e.g., Y chromosomal DNA, RhD gene when the mother is RhD negative) by techniques such as real-time polymerase chain reaction (PCR). However, detection of subtle fetal mutations is difficult due to the overwhelming maternal DNA background. A method combining PCR, base extension reaction, and matrix-assisted laser desorption ionization/time of flight mass spectrometry (MALDI-TOF MS) allowing DNA detection with single base specificity and single DNA molecule sensitivity is described. DNA sequence is amplified by PCR first. Then, a third primer (extension primer) is designed to anneal to the region immediately upstream of the mutation site. Depending on the specific mutation and the ddNTP/dNTP mixtures used, either one or two bases are added to the extension primer to produce two extension products from the wild-type DNA and the mutant DNA. Last, the two extension products are detected by high-throughput MALDI-TOF MS. In addition, with an improved base extension method called single allele base extension reaction, fetal DNA can be robustly detected even when overwhelming maternal background DNA is present. PMID- 18425488 TI - Isolation of cell-free RNA from maternal plasma. AB - The discovery of cell-free RNA in plasma and serum samples provides possibilities for noninvasive prenatal diagnosis. Quantitative alterations of cell-free placental-derived mRNA in the maternal circulation are associated with many pregnancy-related disorders, such as preeclampsia and preterm labor. Obtaining circulating cell-free RNA is the first and often the most critical step for analyzing the placental-derived mRNA in maternal blood. We have compared different protocols for the extraction of cell-free RNA from plasma samples, and we have found the protocol using TRIzol LS reagent (Invitrogen) as lysis buffer combined with RNeasy Mini kit (QIAGEN) to be the optimal method for extracting high-quality cell-free RNA in the highest quantities possible. This method is also amenable to the simultaneous extraction of cell-free RNA from many different samples, by use of the QIAGEN Vacuum Manifold QIAvac 24 Plus. PMID- 18425489 TI - A microarray approach for systematic identification of placental-derived RNA markers in maternal plasma. AB - Circulating fetal RNA in maternal plasma has offered a new approach for noninvasive prenatal diagnosis and monitoring. Circulating fetal RNA markers could potentially be used for all pregnant women without being limited by fetal maternal genetic polymorphisms and fetal gender. Over the past few years, encouraging findings have been reported on the detection and possible clinical applications of circulating fetal RNA. Placental-derived RNA has been shown to be easily detectable in maternal plasma during pregnancy and rapidly cleared after delivery. Such observations suggest that the placenta is an important organ for releasing fetal RNA into maternal plasma. Noninvasive prenatal gene expression profiling of the placenta also has been demonstrated to be feasible by analyzing the circulating placental RNA in maternal plasma. Thus, circulating placental RNA is a potentially useful tool for noninvasive investigation of the placenta. Here, we describe a systematic method for efficient development of new placental specific RNA markers that could be detected in maternal plasma. The method is based on the use of oligonucleotide microarray (Affymetrix, Santa Clara, CA) technology to simultaneously analyze >39,000 RNA transcripts in the placenta. This development has implication for the development of new markers?break for studying disease conditions associated with placental pathology, such as preeclampsia. PMID- 18425490 TI - A novel method to identify syncytiotrophoblast-derived RNA products representative of trisomy 21 placental RNA in maternal plasma. AB - A novel in vitro method is described wherein gene expression profiling is reflective and informative for the way how syncytiotrophoblast cells shed RNA products in vivo in maternal plasma. After controlled denudation, RNA is obtained selectively from the syncytiotrophoblast cells of trisomy 21 placentae. cDNA copies are subsequently analyzed by microarray profiling and cDNA cloning with sequencing. Given the preponderance of 5' mRNA fragments lacking a poly-A tail, the placental RNA products are amplified after polymerase A-mediated tailing by using a method originally designed for small-sized microRNAs. This approach, when combined with cDNA library or microarray expression screening, is a novel in vitro method to screen for syncytiotrophoblast-derived RNA products representative of trisomy 21 placental RNA as present in vivo in maternal plasma. PMID- 18425491 TI - Method for extraction of high-quantity and -quality cell-free DNA from amniotic fluid. AB - Circulating cell-free fetal deoxyribonucleic acids (cffDNAs) are promising biomarkers with various potential clinical applications. Second and third trimester amniotic fluid (AF) is a rich source of cffDNAs. Further improvements to the original protocol for the extraction of cffDNAs from AF supernatant resulted in statistically significant higher yields of high-quality cffDNAs, allowing for a substantial majority of samples to be analyzed with subsequent molecular methods (e.g., comparative genomic hybridization microarrays) to further assess for genetic abnormalities. Several advantages have been realized with the optimized protocol. In addition to an improved yield from a greater proportion of samples compared with the original protocol, the current method, using large silico-membranes, allows for the extraction of cffDNAs from up to 10 samples in <3 h. The replacement of the original lysis buffer eliminates the need for a heating bath during the lysis step, and fewer overall steps are involved in the protocol (e.g., to reduce potential contamination). The improvements in the yield with the current protocol make it possible to augment current standard of care through the analysis of this previously unappreciated source of genetic material. Furthermore, the improvements allow for exploration of widely unknown genetic, pathophysiological, and kinetic issues of cell-free fetal DNA in AF. PMID- 18425492 TI - Detection of new screening markers for fetal aneuploidies in maternal plasma: a proteomic approach. AB - Proteomics has brought with it the hope of identifying novel biomarkers for the fetal aneuploidies. This hope is built on the ability of proteomic technologies, such as two-dimensional gel electrophoresis (2-DE) with immobilized pH gradients combined with protein identification by mass spectrometry (MS). The large dynamic range of plasma necessitates the effective removal of abundant plasma proteins to allow analysis of the lower concentration analytes. There are many factors that make this research very challenging, beginning with standardization of sample collection, consistent sample preparation, and continuing through the entire analytical process. Therefore, reproducible sample complexity reduction methods such as depletions or fractionations are an essential first step in biomarker discovery experiments. For qualitative and quantitative evaluation of the proteome, the fluorescent dye stains offer several advantages over traditional staining methods. The sensitivity of the fluorescent dye stains such as SYPRO Ruby is comparable with that of silver staining and also has a broad dynamic range, which allows accurate protein quantification being compatible with MS methods for protein identification. Despite its limitations for proteome analysis 2-DE is currently the workhorse for proteomics. Taking into account the factors such as cost, availability and ease of use, 2-DE electrophoresis is one of the most apposite approaches toward the methodical characterization of proteomes. PMID- 18425494 TI - Iron and inflammation: cross-talk between pathways regulating hepcidin. PMID- 18425495 TI - Increased susceptibility to endotoxic shock in transgenic rats with endothelial overexpression of kinin B(1) receptors. AB - Two kinin receptors have been described, the inducible B(1) and the constitutive B(2). B(1) receptors are important in cardiovascular homeostasis and inflammation. To further clarify their vascular function, we have generated transgenic rats (TGR(Tie2B(1))) overexpressing the B(1) receptor exclusively in the endothelium. Endothelial cell-specific expression was confirmed by B(1) agonist-induced relaxation of isolated aorta, which was abolished by endothelial denudation of the vessel. This vasodilatation was mediated by nitric oxide (NO) and K(+) channels. TGR(Tie2B(1)) rats were normotensive but, in contrast to controls, reacted with a marked fall in blood pressure and increased vascular permeability after intravenous injection of a B(1) agonist. After lipopolysaccharide treatment, they present a more pronounced hypotensive response and marked bradycardia associated with increased mortality when compared to non transgenic control animals. Thus, the transgenic rats overexpressing kinin B(1) receptors exclusively in the endothelium generated in this study support an important role of this receptor in the vasculature during the pathogenesis of endotoxic shock. PMID- 18425496 TI - HTLV-1 tax-induced NF-kappaB activation is synergistically enhanced by 12-O tetradecanoylphorbol-13-acetate: mechanism and implications for Tax oncogenicity. AB - Nuclear factor kappa B (NF-kappaB) factors regulate a wide range of physiological and oncogenic processes. Normally, these factors are transiently activated by specific external signals which induce their dissociation from inhibitors of kappaB (IkappaB) and subsequent translocation to the nucleus where p65 links to the cyclic adenosine monophosphate response element binding protein (CBP)-p300 and P/CAF coactivators that are essential for its transcriptional activity. The pathogenic potential of human T-cell leukemia virus type 1 (HTLV-1) Tax protein is partly ascribed to its capacity to constitutively activate NF-kappaB factors because constitutive activity of these factors play an important role in the pathophysiology of adult T-cell leukemia (ATL) and tropical spastic paraparesis HTLV-1 associated myelopathy (TSP-HAM). In assessing the possibility of modulating Tax pathogenic potential by external factors, we focused here on 12-O tetradecanoylphorbol-13-acetate (TPA) which is a potent protein kinase C (PKC) activator. There are conflicting reports regarding the effect of TPA and PKC on NF-kappaB. Therefore, we reassessed this issue and also investigated their influence on Tax-mediated activation of these factors. We found that TPA promoted NF-kappaB nuclear translocation and the DNA binding of p65 dimers through PKC activation. However, both TPA and ectopically expressed PKC had only a marginal effect on the transcriptional competence of these dimers, indicating that the DNA binding of such dimers is insufficient by itself for gene activation. Notably, however, both TPA and the ectopic PKC displayed strong synergistic enhancement of the Tax-induced activation of the NF-kappaB transcriptional function. In contrast, TPA and the ectopic PKC only slightly elevated the low activation of the NF-kappaB transcriptional capacity by cytoplasmic Tax mutants, indicating that the nuclear translocation of Tax was essential for this synergism. Subsequent experiments suggested that TPA contributed to this synergism by increasing the pool of free p65 which Tax could link to CBP and elevate, thereby, the amount of a p65-Tax-CBP ternary complex that could bind to NF-kappaB responsive promoters and stimulate their expression. Finally, we demonstrated that this synergism operated also in HTLV-1-infected human T-cells. Earlier reports have shown a close linkage of pathological PKC-activating conditions (e.g., infectious and inflammatory diseases) to certain malignancies. On this ground, the present study suggests that such conditions may enhance the risk for ATL and TSP-HAM in HTLV-1 carriers by increasing the Tax-induced NF-kappaB activation. PMID- 18425497 TI - [In vitro study on the influence of fibrin in cartilage constructs based on PGA fleece materials]. AB - BACKGROUND: The matrix component in autologous chondrocyte implantation plays an important role. In this study the influence of an additional fibrin component in cartilage constructs based on polyglycolide polymers (PGA) was investigated. METHODS: Human chondrocytes of femoral heads were isolated and cultured using a serum-free technique. The cells were seeded on PGA-91 scaffolds with and without an additional fibrin component; the constructs were cultured for 2 weeks in vitro. Besides cell viability, DNA content, pH, aggrecan production, mRNA expression of aggrecan, and collagen types I and II were determined by real-time PCR. Furthermore, cartilage grafts were histologically analyzed. RESULTS: All constructs contained viable, metabolically active cells in the investigated time period. There was no cell proliferation within the graft, and the DNA content was decreased over time. The pH level constantly remained within a physiologic range. The Alcian blue staining of the constructs showed the homogeneous cell distribution and a cell-associated proteoglycan production. Aggrecan concentration in the supernatants of fibrin-containing constructs was significantly lower compared to fibrin-free grafts (-24%), a result that correlated with diminished aggrecan mRNA expression (-80%). mRNA expression of collagen type II increased in the fibrin-free constructs over time and was 57% higher than in the fibrin-containing grafts. The immunohistochemical detection of collagen type II was possible in all constructs. CONCLUSION: Cartilage constructs based on carbohydrate matrices are suitable for matrix-associated chondrocyte implantation. The results of this study suggest a partially inhibitory effect of an additional fibrin component in PGA constructs for chondrogenic differentiation. PMID- 18425498 TI - [The anatomical precontoured Meves compression plate: surgical treatment of clavicular nonunion]. AB - BACKGROUND: Pseudarthroses of the clavicle after fractures of the medial third often present with local pain, compromised shoulder function, or neurovascular symptoms. Reconstruction of normal clavicular anatomy and solid fusion is a prerequisite for good clinical outcome after surgical treatment. In this study, 24 patients with clavicular pseudarthrosis were treated with the anatomical precontoured Meves plate. In 11 patients, additional bone grafting was done. METHOD: Nineteen patients could be reexamined with a mean follow-up of 74.5 months. In all of them, solid fusion was achieved. RESULTS: The Constant score improved from 70.4 points preoperatively up to 82.5 points postoperatively (89.3% age-related). Sixteen patients were satisfied or very satisfied with the operative result. CONCLUSION: In our patients, secure healing of clavicular nonunion was achieved with the anatomical precontoured Meves plate, with good or excellent clinical outcomes. PMID- 18425499 TI - The role of water fittings in intensive care rooms as reservoirs for the colonization of patients with Pseudomonas aeruginosa. AB - OBJECTIVE: To assess the role of the water environment in the Pseudomonas aeruginosa colonization of patients in intensive care units in the absence of a recognized outbreak. DESIGN AND SETTING: Prospective, single-centre study over an 8-week period in two adult ICUs at a university hospital. Environmental samples were taken from the water fittings of rooms once per week, during a 8-week period. Patients were screened weekly for P. aeruginosa carriage. Environmental and humans isolates were genotyped by using pulsed-field gel electrophoresis. RESULTS: P. aeruginosa was detected in 193 (86.2%) of the 224 U-bend samples and 10 of the 224 samples taken from the tap (4.5%). Seventeen of the 123 patients admitted were colonized with P. aeruginosa. Only one of the 14 patients we were able to evaluate was colonized by a clone present in the water environment of his room before the patient's first positive sample was obtained. CONCLUSION: The role of the water environment in the acquisition of P. aeruginosa by intensive care patients remains unclear, but water fittings seem to play a smaller role in non-epidemic situations than expected by many operational hospital hygiene teams. PMID- 18425500 TI - Characterisation of morphology of self-assembled PEG monolayers: a comparison of mixed and pure coatings optimised for biosensor applications. AB - For detection of low concentrations of analytes in complex biological matrices using optical biosensors, a high surface loading with capture molecules and a low nonspecific binding of nonrelevant matrix molecules are essential. To tailor biosensor surfaces in such a manner, poly(ethylene glycols) (PEG) in varying lengths were immobilised covalently onto glass-type surfaces in different mixing ratios and concentrations, and were subsequently modified with three different kinds of receptors. The nonspecific binding of a model protein (ovalbumin, OVA) and the maximum loading of the respective analytes to these prepared surfaces were monitored using label-free and time-resolved reflectometric interference spectroscopy (RIfS). The three different analytes used varied in size: 150 kDa for the anti-atrazine antibody, 60 kDa for streptavidin and 5 kDa for the 15-bp oligonucleotide. We investigated if the mixing of PEG in different lengths could increase the surface loadings of analyte mimicking a three-dimensional matrix as was found using dextrans as sensor coatings. In addition, the effect on the surface loading was investigated with regard to the size of the analyte molecule using such mixed PEGs on the sensor surface. For further characterisation of the surface coatings, polarisation modulation infrared reflection absorption spectroscopy, atomic force microscopy, and ellipsometry were applied. PMID- 18425501 TI - Molecularly imprinted solid-phase extraction and flow-injection chemiluminescence for trace analysis of 2,4-dichlorophenol in water samples. AB - Highly sensitive flow-injection chemiluminescence (CL) combined with molecularly imprinted solid-phase extraction (MISPE) has been used for determination of 2,4 dichlorophenol (2,4-DCP) in water samples. The molecularly imprinted polymer (MIP) for 2,4-DCP was prepared by non-covalent molecular imprinting methods, using 4-vinylpyridine (4-VP) and ethylene glycol dimethacrylate (EGDMA) as the monomer and cross-linker, respectively. 2,4-DCP could be selectively adsorbed by the MIP and the adsorbed 2,4-DCP was determined by its enhancing effect on the weak chemiluminescence reaction between potassium permanganate and luminol. The enhanced CL intensity was linear in the range from 1 x 10(-7) to 2 x 10(-5) g mL( 1). The LOD (S/N = 3) was 1.8 x 10(-8) g mL(-1), and the relative standard deviation (RSD) was 3.0% (n = 11) for 1.4 x 10(-6) g mL(-1). The proposed method had been successfully applied to the determination of 2,4-DCP in river water. PMID- 18425502 TI - Application of matrix solid-phase dispersion to the determination of amitrole and urazole residues in apples by capillary electrophoresis with electrochemical detection. AB - A new method based on matrix solid-phase dispersion (MSPD) extraction was studied for the extraction of amitrole (3-amino-1,2,4-triazole), and its metabolite urazole (3,5-dihydroxy-1,2,4-triazole), in apple samples. The influence of experimental conditions on the yield of the extraction process and on the efficiency of the cleanup step was evaluated. Determination was carried out by capillary electrophoresis (CE) with electrochemical detection, demonstrating the compatibility between MSPD and CE techniques. The method has been successfully applied to different apple varieties. Recoveries in samples spiked at 1.6 and 1.7 microg g(-1) for amitrole and urazole were 88 and 82%, respectively. The limits of detection were 0.4 microg g(-1) for both compounds using electrochemical detection. PMID- 18425503 TI - Molecular iodine selective membrane for iodate determination in salt samples: chemical amplification and preconcentration. AB - A molecular iodine selective membrane has been used for preconcentration of I(2) generated in situ by iodometric reaction of IO(3)(-) with excess I(-) in acidic medium (pH 1-2). This iodometric reaction amplifies the iodine content six times resulting in enhancement of analytical response ranging from three times for molecular methods to six times for elemental methods. The chemical conditions of this iodometric reaction were optimized for quantitative generation and subsequent sorption of I(2) in the membrane samples (96 +/- 3%). The homogeneous transparent membrane was prepared by immobilizing I(2)-complexing polyvinylpyrrolidone (PVP) in the plasticized cellulose triacetate matrix. Four different analytical methods were examined for quantitative determination of IO(3)(-) in iodized salt samples by preconcentrating it as I(2) in the membrane matrix. These methods were: (1) spectrophotometry of the PVP-I(2) complex formed in the membrane matrix, (2) a radiotracer method using I(-) tagged with (131)I radiotracer, (3) instrumental neutron activation analysis (INAA), and (4) energy dispersive X-ray fluorescence (EDXRF) analysis. The IO(3)(-) contents thus determined in the iodized salt samples by the membrane-based radiotracer method were compared with the total iodine determined in salt samples by epithermal instrumental neutron activation analysis (EINAA). The membrane-based method for iodate determination in salt samples has advantages over conventional analytical methods, for example preconcentration and chemical amplification, and is free from interference from anions. PMID- 18425504 TI - Pitch and space maps of skilled cellists: accuracy, variability, and error correction. AB - Based on a newly developed method that combines finger position tracking and spectral analysis of the concurrent acoustic record, we studied the accuracy and variability of pitch performance in eight skilled cellists and the role of acoustic feedback in their performance. The tasks required shifting movements between pairs of notes and separated by various distances (pitch intervals) on a single string at the rate of 1 note/s. The same tasks were performed either using the bow, providing acoustic feedback, or without the bow. Overall, our subjects exhibited a high degree of accuracy in executing tasks when using the bow. When using the bow, two types of variability were observed: (1) trial-to-trial variability: in most subjects the mean fundamental frequency of a single nominal note was significantly different from trial to trial; and (2) within-trial variability. The within-trial variability includes two sub-types: (a) the pitch of a given note changed between notes within a 50-note trial; and (b) within a single note there were positional changes that we hypothesize are attempts by the performer to adjust the fundamental pitch within the note. When acoustic feedback was absent, note distributions were shifted, multimodal, and had large variability; error-correction movements within a single note also significantly decreased, indicating that the stability and precision of the motor map depends on constant re-calibration and updating by acoustic information. Our results suggest that a performer's intonation should not be viewed as a fixed entity implied by the score but as a sample from a statistical distribution. PMID- 18425505 TI - Neural activation in cognitive motor processes: comparing motor imagery and observation of gymnastic movements. AB - The simulation concept suggested by Jeannerod (Neuroimage 14:S103-S109, 2001) defines the S-states of action observation and mental simulation of action as action-related mental states lacking overt execution. Within this framework, similarities and neural overlap between S-states and overt execution are interpreted as providing the common basis for the motor representations implemented within the motor system. The present brain imaging study compared activation overlap and differential activation during mental simulation (motor imagery) with that while observing gymnastic movements. The fMRI conjunction analysis revealed overlapping activation for both S-states in primary motor cortex, premotor cortex, and the supplementary motor area as well as in the intraparietal sulcus, cerebellar hemispheres, and parts of the basal ganglia. A direct contrast between the motor imagery and observation conditions revealed stronger activation for imagery in the posterior insula and the anterior cingulate gyrus. The hippocampus, the superior parietal lobe, and the cerebellar areas were differentially activated in the observation condition. In general, these data corroborate the concept of action-related S-states because of the high overlap in core motor as well as in motor-related areas. We argue that differential activity between S-states relates to task-specific and modal information processing. PMID- 18425506 TI - Do synergies decrease force variability? A study of single-finger and multi finger force production. AB - We tested a hypothesis that force production by multi-finger groups leads to lower indices of force variability as compared to similar single-finger tasks. Three experiments were performed with quick force production, steady-state force production under visual feedback, and steady-state force production without visual feedback. In all experiments, a range of force levels was used computed as percentages of the maximal voluntary contraction force for each involved finger combination. Force standard deviation increased linearly with force magnitude across all three experiments and all finger combinations. There were modest differences between multi-finger and single-finger tasks in the indices of force variability, significant only in the tasks with steady-state force production under visual feedback. When fingers acted in groups, each finger showed significantly higher force variability as compared to its single-finger task and as compared to the multi-finger group as a whole. Fingers that were not instructed to produce force also showed close to linear relations between force standard deviation and force magnitude. For these fingers, indices of force variability were much higher as compared to those computed for the forces produced by instructed fingers. We interpret the findings within a feed-forward scheme of multi-finger control with two inputs only one of which is related to the explicit task. The total force variability reflects variability in only the task-related component, while variability of the finger forces is also due to variability of the component that is not related to the task. The findings tentatively suggest that total force variability originates at an upper level of the control hierarchy in accordance to the Weber-Fechner law rather than reflects a "neural noise" at the segmental level. PMID- 18425507 TI - Delayed discrimination of spatial frequency for gratings of different orientation: behavioral and fMRI evidence for low-level perceptual memory stores in early visual cortex. AB - The concept of perceptual memory refers to the neural and cognitive processes underlying the storage of specific stimulus features such as spatial frequency, orientation, shape, contrast, and color. Psychophysical studies of perceptual memory indicate that observers can retain visual information about the spatial frequency of Gabor patterns independent of the orientation with which they are presented. Compared to discrimination of gratings with the same orientation, reaction times to orthogonally oriented gratings, however, increase suggesting additional processing. Using event-related fMRI we examined the pattern of neural activation evoked when subjects discriminated the spatial frequency of Gabors presented with the same or orthogonal orientation. Blood-oxygen level dependent BOLD fMRI revealed significantly elevated bilateral activity in visual areas (V1, V2) when the gratings to be compared had an orthogonal orientation, compared to when they had the same orientation. These findings suggest that a change in an irrelevant stimulus dimension requires additional processing in primary and secondary visual areas. The finding that the task-irrelevant stimulus property (orientation) had no significant effect on the prefrontal and intraparietal cortex supports a model of working memory in which discrimination and retention of basic stimulus dimensions is based on low-level perceptual memory stores that are located at an early stage in the visual process. Our findings suggest that accessing different stores requires time and has higher metabolic costs. PMID- 18425508 TI - Evaluation of complexed NO reduction mechanism in a chemical absorption biological reduction integrated NO(x) removal system. AB - Biological reduction of nitric oxide (NO) from Fe(II) ethylenediaminetetraacetic acid (EDTA)-NO to dinitrogen (N(2)) is a core process for the continual nitrogen oxides (NO(x)) removal in the chemical absorption-biological reduction integrated approach. To explore the biological reduction of Fe(II)EDTA-NO, the stoichiometry and mechanism of Fe(II)EDTA-NO reduction with glucose or Fe(II)EDTA as electron donor were investigated. The experimental results indicate that the main product of complexed NO reduction is N(2), as there was no accumulation of nitrous oxide, ammonia, nitrite, or nitrate after the complete depletion of Fe(II)EDTA-NO. A transient accumulation of nitrous oxide (N(2)O) suggests reduction of complexed NO proceeds with N(2)O as an intermediate. Some quantitative data on the stoichiometry of the reaction are experimental support that reduction of complexed NO to N(2) actually works. In addition, glucose is the preferred and primary electron donor for complexed NO reduction. Fe(II)EDTA served as electron donor for the reduction of Fe(II)EDTA-NO even in the glucose excessive condition. A maximum reduction capacity as measured by NO (0.818 mM h(-1)) is obtained at 4 mM of Fe(II)EDTA-NO using 5.6 mM of glucose as primary electron donor. These findings impact on the understanding of the mechanism of bacterial anaerobic Fe(II)EDTA-NO reduction and have implication for improving treatment methods of this integrated approach. PMID- 18425509 TI - Insertion sequence-based cassette PCR: cultivation-independent isolation of gamma hexachlorocyclohexane-degrading genes from soil DNA. AB - gamma-Hexachlorocyclohexane (gamma-HCH) is a highly chlorinated pesticide that has caused serious environmental problems. Based on the frequently observed association of insertion sequence IS6100 with lin genes for gamma-HCH degradation in several gamma-HCH-degrading bacterial strains isolated to date, DNA fragments flanked by two copies of IS6100 were amplified by nested polymerase chain reaction (PCR) technique using a DNA sample extracted from soil contaminated with HCH. Four distinct DNA fragments with sizes of 6.6, 2.6, 1.6, and 1.3 kb were obtained, three of which carried lin genes: the 6.6-kb fragment carried linD and linE as well as linR; the 2.6-kb fragment showed a truncated form of linF; and the 1.6-kb fragment carried linB. Our approach, named as insertion sequence (IS) based cassette PCR, was successful in the isolation of the lin genes from HCH contaminated soil without cultivation of host cells and is applicable for the culture-independent isolation of other functional genes bordered by other IS elements. PMID- 18425510 TI - High-yield growth and magnetosome formation by Magnetospirillum gryphiswaldense MSR-1 in an oxygen-controlled fermentor supplied solely with air. AB - Magnetotactic bacteria are difficult to grow under defined conditions in culture, which has presented a major obstacle to commercial application of magnetosomes. We studied the relationships among the cell growth, magnetosome formation, dissolved oxygen concentration (DO), and the ability to supply oxygen to the cells. Mass culture of Magnetospirillum gryphiswaldense MSR-1 for the production of magnetosomes was established in a 42-L fermentor under the following conditions: (1) sterile air was the sole gas supplied in the fermentor, and DO could be regulated at any level below 10% saturation by cascading the stir rate to DO, (2) to resolve the paradoxical situation that the cell growth requires higher DO whereas magnetosome formation requires low DO below the detectable range of regular oxygen electrode, DO was controlled to optimal level using the change of cell growth rate, rather than reading from the highly sensitive oxygen electrode, as the signal for determining appropriate DO, and (3) timing and rate of supplying the substrates were determined by measuring cell density and Na lactate concentration. Under these conditions, cell density (OD565) of strain MSR 1 reached 7.24 after 60-h culture in a 42-L fermentor, and cell yield (dry weight) was 2.17 g/L, the highest yield so far being reported. The yield of magnetosomes (dry weight) was 41.7 mg/L and 16.7 mg/L/day, which were 2.8 and 2.7 times higher than the previously reported yields. PMID- 18425511 TI - SPECT imaging of D2 dopamine receptors and endogenous dopamine release in mice. AB - PURPOSE: The dopamine D(2) receptor (D2R) is important in the mediation of addiction. [(123)I]iodobenzamide (IBZM), a SPECT ligand for the D2R, has been used for in vivo studies of D2R availability in humans, monkeys, and rats. Although mouse models are important in the study of addiction, [(123)I]IBZM has not been used in mice SPECT studies. This study evaluates the use of [(123)I]IBZM for measuring D2R availability in mice. METHODS: Pharmacokinetics of [(123)I]IBZM in mice were studied with pinhole SPECT imaging after intravenous (i.v.) injection of [(123)I]IBZM (20, 40, and 70 MBq). In addition, the ability to measure the release of endogenous dopamine after amphetamine administration with [(123)I]IBZM SPECT was investigated. Thirdly, i.v. administration, the standard route of administration, and intraperitoneal (i.p.) administration of [(123)I]IBZM were compared. RESULTS: Specific binding of [(123)I]IBZM within the mouse striatum could be clearly visualized with SPECT. Peak specific striatal binding ratios were reached around 90 min post-injection. After amphetamine administration, the specific binding ratios of [(123)I]IBZM decreased significantly (-27.2%; n = 6; p = 0.046). Intravenous administration of [(123)I]IBZM led to significantly higher specific binding than i.p. administration of the same dose. However, we found that i.v. administration of a dose of 70 MBq [(123)I]IBZM might result in acute ethanol intoxication because ethanol is used as a preparative aid for the routine production of [(123)I]IBZM. CONCLUSIONS: Imaging of D2R availability and endogenous dopamine release in mice is feasible using [(123)I]IBZM single pinhole SPECT. Using commercially produced [(123)I]IBZM, a dose of 40 MBq injected i.v. can be recommended. PMID- 18425512 TI - Concordance between results of somatostatin receptor scintigraphy with 111In-DOTA DPhe 1-Tyr 3-octreotide and chromogranin A assay in patients with neuroendocrine tumours. AB - PURPOSE: Somatostatin receptor scintigraphy (SRS) and chromogranin A (CgA) assay have successfully been implemented in the clinical work-up and management of neuroendocrine tumour (NET) patients. However, there is still a lack of studies comparing results in these patients. Our aim was to compare directly in NET patients SRS and CgA assay results with special regard to tumour features such as grade of malignancy, primary origin, disease extent and function. METHODS: One hundred twenty consecutive patients with histological confirmed NETs were investigated with (111)In-DOTA-DPhe(1)-Tyr(3)-octreotide ((111)In-DOTA-TOC) SRS and CgA immunoradiometric assay. Tumours were classified by cell characteristics [well-differentiated NETs, well-differentiated neuroendocrine carcinomas, poorly differentiated neuroendocrine carcinomas (PDNECs)], primary origin (foregut, midgut, hindgut, undetermined), disease extent (limited disease, metastases, primary tumour and metastases) and functionality (secretory, nonsecretory). RESULTS: SRS was positive in 107 (89%) patients; CgA levels were increased in 95 (79%) patients. Overall, concordance between SRS and CgA results was found in 84 patients. Positive SRS but normal CgA level were found in 24 patients, with higher prevalence (p<0.05) in patients with nonsecretory tumours. Conversely, negative SRS but CgA level increased were seen in 12 patients, with higher proportion (p<0.05) in patients with PDNECs and tumours of hindgut origin. CONCLUSIONS: Overall, (111)In-DOTA-TOC SRS proved to be more sensitive than CgA in NETs patients. Tumour differentiation, disease extent and presence of liver metastases impact both SRS and CgA results, whereas nonsecretory activity is a negative predictor of only CgA increase. PDNECs and hindgut origin of tumours predispose to discrepancies with negative SRS but increased CgA levels. PMID- 18425513 TI - Quantitative relationship between coronary vasodilator reserve assessed by 82Rb PET imaging and coronary artery stenosis severity. AB - PURPOSE: The relationship between myocardial blood flow (MBF) and stenosis severity has been determined previously using cyclotron-produced radiotracers such as (15)O-H(2)O and (13)N-ammonia. An attractive alternative to overcome the limitations related to the use of cyclotron might be to use the generator produced (82)Rb as a flow tracer. The current study was undertaken to investigate the relationship between MBF and coronary vasodilator reserve (CVR) as measured by (82)Rb positron emission tomography (PET) and the percent diameter stenosis as defined by quantitative coronary arteriography. METHODS: We prospectively evaluated 22 individuals: 15 patients (60 +/- 11 years of age) with angiographically documented coronary artery disease (CAD) and seven age-matched (56 +/- 9 years) asymptomatic individuals without risk factors for CAD. Dynamic (82)Rb PET was performed at rest and after dipyridamole vasodilation. MBF, CVR and an index of 'minimal coronary resistance' (MCR) were assessed in each of the three main coronary territories. RESULTS: Rest and stress MBF in regions subtended by vessels with less than 50% diameter stenosis was similar to that of the individuals with no risk factors for CAD. As a result, CVR was also similar in the two groups (1.9, interquartile [IQ] range from 1.7 to 2.7 vs. 2.2, IQ range from 2 to 3.4 respectively, p = 0.09). CVR successfully differentiated coronary lesions with stenosis severity 70% to 89% from those with 50% to 69% stenosis (1, IQ range from 1 to 1.3 vs. 1.7, IQ range from 1.4 to 2), respectively, p = 0.001. In addition, hyperaemic MBF (r (2) = 0.74, p < 0.001), CVR (r (2) = 0.69, p < 0.001) and MCR (r (2) = 0.78, p < 0.001) measurements were inversely and non-linearly correlated to the percent diameter stenosis on angiography. CONCLUSION: MBF and CVR are inversely and non-linearly correlated to stenosis severity. Quantitative (82)Rb PET can be a clinically useful tool for an accurate functional assessment of CAD. PMID- 18425514 TI - Clinical use of dopamine transporter imaging in movement disorders: benefits of appropriate use. PMID- 18425515 TI - Role of cardiac nuclear imaging in the management of critically ill patients. PMID- 18425516 TI - Exploratory evaluation of two-dimensional and three-dimensional methods of FDG PET quantification in pediatric anaplastic astrocytoma: a report from the Pediatric Brain Tumor Consortium (PBTC). AB - PURPOSE: The rationale of this study was to investigate the feasibility of three dimensional (3D) methods to analyze (18)F-fluoro-deoxy-glucose (FDG) uptake in children with anaplastic astrocytoma (AA) in a multi-institutional trial, to compare 3D and two-dimensional (2D) methods and explore data associations with progression-free survival (PFS). METHODS: 3D tumor volumes from pretreatment MR images (fluid attenuation inversion recovery and postgadolinium) of children with recurrent AA on a phase I trial of imatinib mesylate were co-registered to FDG positron emission tomography (PET) images. PET data were normalized. Four metrics were defined: the maximum ratio (maximum pixel value within the 3D tumor volume, normalized), the total ratio (cumulative pixel values within the tumor volume, normalized) and tumor mean ratio (total pixel value divided by volume, normalized). 2D analysis methods were compared. Cox proportional hazards models were used to estimate the association between these methods and PFS. RESULTS: Strongest correlations between 2D and 3D methods were with analyses using postcontrast T1 images for volume of interest (VOI). The analyses suggest 3D maximum tumor and mean tumor ratios, whether normalized by gray matter or white matter, were associated with PFS. CONCLUSIONS: This study of a series of pretreatment AA patients suggests that 3D PET methods using VOIs based on postcontrast T1 correlate with 2D methods and are related to PFS. These methods yield an estimate of metabolically active tumor burden and may add prognostic information after tumor grade is determined. Future rigorous multi-institutional protocols with larger numbers of patients will be required for validation. PMID- 18425517 TI - Regulation of expression of rice thaumatin-like protein: inducibility by elicitor requires promoter W-box elements. AB - Rice thaumatin-like protein (Rtlp1) is a high-molecular-weight antimicrobial pathogenesis-related protein that plays a role in plant stress response. This study examines transcriptional regulation of Rtlp1 using wild type and transgenic rice plants carrying a beta-glucuronidase (GUS) reporter gene driven by the Rtlp1 promoter (pRtlp1GUS). The Rtlp1 promoter is induced within 6 h after infection with rice blast fungus (Magnaporthe grisea). The Rtlp1 promoter is also induced by salicylic acid (SA), methyl jasmonate (MeJA), wounding or an elicitor from rice blast fungus. The function of the pRtlp1GUS reporter gene was analyzed by deletion mapping and transient expression assays in cell culture. A 120 bp truncated fusion construct with six W-boxes (5'-TGAC-3') demonstrated a strong dose-dependent elicitor-response. These results suggest that W-box elements are required for the response of the Rtlp1 promoter to fungal elicitors. PMID- 18425518 TI - Performance of FDG-PET/CT for diagnosis of recurrent uterine cervical cancer. AB - The purpose is to evaluate the accuracy of integrated FDG-PET/CT, compared with PET alone, for diagnosis of suspected recurrence of uterine cervical cancer. Fifty-two women who had undergone treatment for histopathologically proven cervical cancer received PET/CT with suspected recurrence. PET-alone and integrated PET/CT images were evaluated by two different experienced radiologists by consensus for each investigation. A final diagnosis was confirmed by histopathology, radiological imaging, and clinical follow-up for over 1 year. Patient-based analysis showed that the sensitivity, specificity, and accuracy of PET/CT were 92.0% (23/25), 92.6% (25/27), and 92.3% (48/52), respectively, while for PET, the corresponding figures were 80.0% (20/25), 77.8% (21/27), and 78.8% (41/52), respectively. PET/CT resolved the false-positive PET results due to hypermetabolic activity of benign/inflammatory lesions and physiological variants, and was able to detect lung metastasis, local recurrence, peritoneal dissemination, para-aortic lymph node metastasis, and pelvic lymph node metastasis missed by PET alone. However, tiny local recurrence and lymph node metastasis could not be detected even by PET/CT. FDG-PET/CT is a useful complementary modality for providing good anatomic and functional localization of sites of recurrence during follow-up of patients with cervical cancer. PMID- 18425519 TI - MR imaging of thrombi using EP-2104R, a fibrin-specific contrast agent: initial results in patients. AB - This study was an initial phase II trial in humans of molecular magnetic resonance (MR) imaging for improved visualization of thrombi in vessel territories potentially responsible for stroke using a new fibrin-specific contrast agent (EP-2104R). Eleven patients with thrombus in the left ventricle (n = 2), left or right atrium (n = 4), thoracic aorta (n = 4) or carotid artery (n = 1) as verified by an index examination (ultrasound, computed tomograpy, or conventional MR) were enrolled. All MR imaging was performed on 1.5 T whole-body MR-system using an inversion-recovery black-blood gradient-echo sequence. The same sequence was performed before and 2-6 h after low-dose intravenous administration of 4 mumol/kg EP-2104R. Two investigators assessed image quality and signal amplification. Furthermore, contrast-to-noise ratios (CNR) between the clot and the blood pool/surrounding soft tissue before and after administration of the contrast agent were compared using Student's t-test. MR imaging and data analysis were successfully completed in 10 patients. No major adverse effects occurred. On enhanced images, thrombi demonstrated high signal amplification, typically at the clot surface, with a significantly increased contrast in comparison to the surrounding blood pool and soft tissue (CNR for clot vs. blood pool, unenhanced and enhanced: 6 +/- 8 and 29 +/- 14; CNR for clot vs. soft tissue, unenhanced and enhanced: 0 +/- 4 and 21 +/- 13; P < 0.01 for both comparisons). EP-2104R allows for molecular MR imaging of thrombi potentially responsible for stroke. High contrast between thrombus and surrounding blood and soft tissues can be achieved with enhanced imaging. PMID- 18425520 TI - CsI-detector-based dual-exposure dual energy in chest radiography for lung nodule detection: results of an international multicenter trial. AB - To assess both sensitivity and specificity of digital chest radiography alone and in conjunction with dual-exposure dual-energy chest radiography for the detection and classification of pulmonary nodules. One hundred patients with a total of 149 lung nodules (3-45 mm; median, 11 mm) confirmed by CT were included in this study. Dual-exposure dual-energy chest radiographies of each patient were obtained using a CsI detector system. Experienced board-certified chest radiologists from four different medical centers in Europe reviewed standard chest radiographs alone and in conjunction with dual-energy images blinded and in random order. The reviewers rated the probability of presence, calcification and malignancy of all lung nodules on a five-point rating scale. Lesions detected were identified by applying a specific coordinate system to enable precise verification by the study leader. A receiver-operating characteristic (ROC) analysis was performed. In addition to the 149 true-positive CT proven lesions, 236 false-positive lung nodules were described in digital chest radiographies in conjunction with dual-energy chest radiographies. The cumulative sensitivity of chest radiography in conjunction with dual energy was 43%, specificity was 55%. For digital radiography alone, sensitivity was 35% and specifity was 83%. For the dual energy system, positive predictive value was 58%, and negative predictive value was 66% compared to the digital radiography with a positive predictive value of 59% and a negative predictive value of 65%. Areas under the curve in a ROC analysis resulted in 0.631 (95% confidence interval =0.61 to 0.65) for radiography with dual energy and 0.602 (95% confidence interval =0.58 to 0.63) for digital radiography alone. This difference was not statistically significant. For the detection of lesion calcification or the determination of malignancy, ROC analysis also failed to show significant differences. CsI-based flat-panel dual exposure dual-energy imaging added to standard chest radiography did not show statistically significant improvement for the detection of pulmonary nodules, nor the identification of calcifications, nor the determination of malignancy. PMID- 18425521 TI - Lingual mucosal graft substitution urethroplasty for anterior urethral strictures: our technique of graft harvesting. AB - OBJECTIVE: To describe our technique of lingual mucosal graft harvesting for substitution urethroplasty and the complications encountered at the donor site. METHODS: Twenty-eight patients who underwent lingual mucosal graft urethroplasty between May 2006 and March 2007 were included in this study. TECHNIQUE: The site of the graft harvest is the lateral mucosal lining of the tongue. Graft harvesting is started from the posterior landmark of the graft on the left side of the tongue. It is continued across the tip of the tongue to the other side if lengthier graft is required. The graft harvesting site is simultaneously closed with continuous running suture using 4-0 polyglactin suture to achieve immediate and good homeostasis. RESULTS: Mean duration of follow up was 4.2 months. Average length of harvested graft was 6.5 cm. Mean harvesting time was 18 min. At the first postoperative day, 92% patients experienced pain at donor site and 24% had slurring of speech. By third postoperative day, >70% were free of pain and four had slurring of speech. By sixth postoperative day, none of the patient suffered pain. All the patients were able to resume oral fluid within 24 h, eat soft solid diet in 48-72 h and return to normal diet after 4-5 days of surgery. No patient suffered from difficulty in opening the mouth, salivation disturbances, peri-oral numbness or difficulty in protrusion of tongue. CONCLUSION: Lingual mucosal graft harvesting is a simple procedure, provides lengthy graft and is associated with minimal donor site complications. PMID- 18425522 TI - Arteriovenous crossing sheathotomy versus intravitreal triamcinolone acetonide injection for treatment of macular edema associated with branch retinal vein occlusion. AB - PURPOSE: To compare the functional and anatomical outcomes of arteriovenous (AV) sheathotomy and intravitreal triamcinolone acetonide (IVTA) injection in the treatment of macular edema associated with branch retinal vein occlusion (BRVO). METHODS: Forty eyes of 40 patients with macular edema secondary to BRVO were randomized into two treatment groups. A total of 20 patients received AV sheathotomy (sheathotomy group), and the second group of 20 patients was treated with IVTA (IVTA group). Early Treatment Diabetic Retinopathy Study (ETDRS) visual acuity (VA) score, total macular volume measured, and foveal thickness by 3rd generation optical coherence tomography (OCT3) were evaluated as main outcome measurements. RESULTS: The average changes in ETDRS scores, total macular volumes, and foveal thicknesses compared to baseline values, were significant 3 months and 6 months after treatment in both groups (P < 0.05, paired t-test), but only the IVTA group showed significant improvements 1 month after treatment. The between-group differences in average ETDRS score, total macular volume, and foveal thickness changes were significantly better at 1 month after treatment in the ITVA group (P = 0.026, P < 0.001, P = 0.001, respectively, Student's t-test), at which time IVTA patients had better vision and anatomical outcomes than did those in the sheathotomy group. CONCLUSIONS: After either AV sheathotomy or IVTA treatment, patients with macular edema secondary to BRVO showed similar functional and anatomical outcomes 6 months later. When the cost and the risks of vitreoretinal surgery are considered, IVTA treatment may be a better treatment option, as the drug yields better short-term outcomes. PMID- 18425523 TI - Persisent ocular hypertension following intravitreal ranibizumab. AB - BACKGROUND: To describe the occurrence of ocular hypertension in four patients following injection of ranibizumab intravitreally. METHODS: Case series. RESULTS: Four patients had high intraocular pressure after intravitreal ranibizumab 0.5 mg. Ocular hypertension occurred 1 month after the second ranibizumab injection in patients 1 and 3, and 1 month after the first ranibizumab in patient 2. In patient 4, it occurred several hours after the first ranibizumab injection. In all patients, the IOP increase was sustained across several visits, requiring control with topical glaucoma therapy, and in two cases the addition of a systemic carbonic anhydrase inhibitor. None of the patients had a previous history of glaucoma, ocular hypertension or IOP asymmetry and the IOP was as high as 30, 34, 46, and 50 mmHg in the four patients. CONCLUSION: Severe and sustained ocular hypertension may occur after intravitreal ranibizumab. Although the mechanism of the pressure rise is unknown, all eyes in our series were controlled with medical therapy. PMID- 18425524 TI - Changes in retinal thickness are correlated with alterations of electroretinogram in eyes with central retinal artery occlusion. AB - BACKGROUND: We investigated the relationship between the retinal thickness and electroretinogram (ERG) components in patients with central retinal artery occlusion (CRAO). METHODS: The optical coherence tomographic (OCT) images and ERGs of the nine patients (six men and three women; mean age, 61.8 years) were retrospectively analyzed. The thickness of the inner and outer retinal layers at 1 and 2 mm nasal and temporal to the fovea was measured in the horizontally scanned OCT images. The ratio of the inner layer thickness/sensory retinal thickness (IT/ST ratio) was calculated. The amplitudes of the a- and b-waves of the mixed rod-cone ERGs and the photopic negative response (PhNR) of the photopic ERGs were analyzed. The ratio of the amplitude of each component in the affected eye to that of the healthy fellow eye (a/f ratio) was calculated. RESULTS: In the chronic phase (1 to 8 months after onset, eight eyes), the inner layer was significantly thinner than that in the acute phase (P = 0.0147, 0.0076, 0.002, and 0.0003 for 2 mm nasal, 1 mm nasal, 1 mm temporal, and 2 mm temporal respectively, within 5 days of onset, six eyes), while the thickness of outer layer was not significantly changed. The ERGs were recorded 6.4 +/- 1.5 days after the onset of CRAO. The median of the a/f ratio was 0.84 in the a-wave, 0.56 in the b-wave, and 0.27 in the PhNR. The IT/ST in the chronic phase was positively correlated with the a/f ratio of the amplitude of the PhNR. CONCLUSIONS: Measurement of retinal thickness by OCT can be useful for monitoring the changes following CRAO. The correlation between the retinal thickness, especially inner layer thickness, and the ERG components was determined, suggesting that the PhNR in the acute phase might be a good indicator for predicting the thinning of the damaged retina in the chronic phase. PMID- 18425525 TI - Posterior capsulorhexis combined with optic buttonholing: an alternative to standard in-the-bag implantation of sharp-edged intraocular lenses? A critical analysis of 1000 consecutive cases. AB - BACKGROUND: Current after-cataract prevention relies on optimizing the natural barrier effect of the optic rim against lens epithelial cell (LEC) migration. However, deficiencies in circumferential capsular bag closure caused by the intraocular lens (IOL) haptic or delayed secondary re-division of the fused capsules by Soemmering s ring formation lead to primary or secondary barrier failure. Consequently, surprisingly high posterior laser capsulotomy rates have been reported long-term, even with optimal capsular surgery and the most widespread hydrophobic acrylic IOLs, considered to be the most advanced. Intraoperative removal of the central posterior capsule has been shown to be effective in further reducing LEC immigration. However, efficacy has turned out to be limited because of the propensity of LECs to use the posterior optic surface as an alternative scaffold. MATERIAL AND METHODS: TECHNIQUE: in pediatric cataract surgery, buttoning-in of the optic into an adequately-centered posterior capsulorhexis opening has been described previously. This technique was further elaborated and applied as the standard technique in a large series of adult eyes. In general, the diameter aimed at was 5-6 mm for the anterior, and 4 5 mm for the posterior capsulorhexis. Between September 2004 and June 2007, 1000 consecutive cases have thus been performed and systematically evaluated. One hundred and fifty eyes additionally underwent extensive anterior LEC abrasion. Another sub-series investigated the option of further reducing capsular fibrosis by creating an anterior capsulorhexis larger than the optic. EVALUATION: special scrutiny was applied to detect postoperative vitreous entrapment. Regeneratory and fibrotic after-cataract formation were both meticulously followed-up. Postoperative pressure course, anterior segment inflammation, macular thickness and morphology, as well as axial optic stability and optic centration, were evaluated in intraindividual comparison studies. RESULTS: A low rate of vitreous complications was found, which can be avoided by appropriate surgery. Vitreous entanglement occurred in six eyes, and vitreous herniation after PPCCC over sizing in two. In three, anterior vitrectomy was performed. There was only one single case of retinal detachment-supposedly unrelated to the technique itself and no case of cystoid macular edema. Retro-optical regenerate formation was completely abolished, while fibrosis was drastically reduced by the posterior capsule sandwiched in between the anterior LEC layer on the backside of the anterior capsule and the anterior optic surface, thereby blocking contact mediated myofibroblastic LEC transdifferentiation. Additional capsular polishing further reduced residual fibrosis emerging from the anterior capsule contacting the optic adjacent to the haptic junction, as well as regeneratory LEC re proliferation on the posterior capsule overlying the optic. Postoperative pressure course was almost identical to that found after standard in-the-bag implantation of the IOL, as was flare, and macular thickness and morphology. As opposed to bag-fixated IOLs, no axial movement of the optic was detected. IOL optics always perfectly centered even when the capsular opening was not optimally centered. Due to the exquisite stretchability and elasticity of the posterior capsule, the 6-mm IOL optic could safely be buttoned-in in a posterior capsulorhexis of 4 mm and smaller. CONCLUSIONS: Posterior optic buttonholing (POBH) is a safe and effective technique which not only excludes retro-optical opacification, but also withholds capsular fibrosis by obviating direct contact between the anterior capsular leaf and the optic surface. Anterior LEC abrasion significantly reduced both the residual fibrosis and regeneratory LEC proliferation. Apart from pediatric cataract, POBH is currently recommended for eyes with pseudoexfoliation syndrome, high axial myopia, peripheral retinal disease, and multifocal IOL implantation. Toric IOLs and magnet-driven accommodative IOL systems are other potential applications. Generally, POBH holds promise for becoming a routine alternative to standard in-the-bag IOL implantation in the future. PMID- 18425526 TI - Tracking of human cells in mice. AB - Tracking and tracing of transplanted cells in mice is required in many fields of research. Examples are transplantation of stem cells into organs of mice to study their differentiation capacity and injection of tumor cells to examine metastatic behavior. In the present study we tested the lipid dye CM-DiI and red fluorescent nanoparticles Qdot655 for their applicability in tagging and tracing of human cells in mice. Labeling of different cell types, including MCF-7 human breast cancer cells, human cord blood derived cells, human NeoHep cells and human hepatopancreatic precursor cells, is technically easy and did not compromise further cell culture. After transplantation of CM-DiI or Qdot655 marked cells, red fluorescent structures could be detected already in unprocessed paraffin slices of the studied organs, namely liver, lung, pancreas, kidney, spleen and bone marrow. Next, we examined whether the red fluorescent structures represent the transplanted human cells. For this purpose, we established an in situ hybridization (ISH) technique that allows clear-cut differentiation between human and murine nuclei, based on simultaneous hybridization with human alu and mouse major satellite (mms) probes. We observed a high degree of coincidence between CM DiI-marked cells and alu positive nuclei. However, also some mms positive cells contained CM-DiI, suggesting phagocytosis of the transplanted CM-DiI-marked cells. The degree of such CM-DiI-positive mouse cells depended on the cell type and route of administration. From a technical point of view it was important that CM-DiI-positive structures in paraffin slices remained fluorescent also after ISH. In contrast, Qdot655 positive structures faded during further staining procedures. In conclusion, marking of cells with CM-DiI or Qdot655 prior to transplantation facilitates recovery of human cells, since a high fraction of positive structures in the host's tissue originate from the transplanted cells. However, CM-DiI or Qdot655 positive staining of individual cells in transplanted tissues is not sufficient to prove their human origin. Additional procedures, such as ISH with alu-probes, are essential, when characterizing individual cells. PMID- 18425527 TI - Paternal work stress and prolonged time to pregnancy. AB - OBJECTIVE: The aim of this study was to explore an association between psychosocial stress at work in married men and their spouses' prolonged time to pregnancy (TTP). METHODS: All married male workers of a large Korean petrochemical enterprise and their wives fulfilling the selection criteria were included. Main selection criteria were lack of use of contraceptives and experienced pregnancy in recent past. Data were available from 322 couples. Psychosocial stress at work was measured by the effort-reward imbalance questionnaire. Prolonged TTP was measured by the "TTP questionnaire". RESULTS: After adjustment for confounding effects of demographic and life-style characteristics and benzene exposure, delayed TTP, defined by frequency of first cycle pregnancy, was associated with one standard deviation (SD) increase of the effort-reward ratio in the chronically stressed group of married men (OR = 0.47; 95% CI = 0.22-0.99) in logistic regression analysis. A similar, but somewhat weaker effect, was found for the overall group (OR = 0.67; 95% CI = 0.47-0.94). CONCLUSIONS: Paternal stress at work, as measured by effort-reward imbalance, seemed to be associated with a decreased number of conceptions in the first menstrual cycle. PMID- 18425528 TI - Levels of 1-hydroxypyrene, symptoms and immunologic markers in vulcanization workers in the southern Sweden rubber industries. AB - OBJECTIVES: The aim of this study was to determine urinary 1-hydroxypyrene (1-HP) levels in contemporary Swedish vulcanization workers and in controls. These levels were used as an index substance for vulcanization fumes, as well as a biomarker for polycyclic aromatic hydrocarbons (PAHs). The risk of symptoms and changed levels of immunologic markers were investigated in relation to the 1-HP levels. METHODS: Included in the study were 163 exposed workers and 106 controls. Medical and occupational histories were obtained by structured interviews. Symptoms were recorded and immunologic markers analysed in blood by routine analysis methods. Levels of 1-HP were determined by liquid chromatography and fluorescence detection. RESULTS: The highest levels of 1-HP were found among exposed workers using injection and compression vulcanization and lower levels were found among exposed workers vulcanizing with salt bath, hot air, microwaves or fluid-bed. Compared to controls, exposed workers had increased risks of eye symptoms, nosebleeds, burning and dry throat, hoarseness, severe dry cough, nausea and headache. Furthermore, exposed workers had elevated levels of neutrophils and total IgG (immunoglobulin subclass G). However, only for severe dry cough an evident exposure-response relationship with urinary 1-HP levels was found. CONCLUSIONS: This work clearly shows increased levels of urinary 1-HP in Swedish vulcanization workers. Furthermore, it demonstrates an increased risk of several symptoms and elevated levels of some immunologic markers in these workers. However, no obvious exposure-response relationships were found. PMID- 18425529 TI - Effects of age on neuromuscular knee joint control. AB - Age-related changes in neuromuscular function may have adverse effects on hamstring reflex activity and may impair knee joint stability. The aim of this study was to investigate whether increasing age affects hamstring short latency responses (SLR), medium latency responses (MLR), and anterior tibial translation. In 40 healthy subjects ranging from 20 to 70 years of age we assessed hamstring muscle latencies and integrals for both hamstring reflex components (SLRs and MLRs) as well as anterior tibial translation in order to quantify knee joint stability in response to induced tibial translation during stance. The results showed no significant differences within the subject population in hamstring SLR and MLR latencies and integrals or anterior tibial translation. More precisely, regression analysis did not reveal any correlation between age and the aforementioned parameters. Our findings suggest that functional knee stability in terms of anterior tibial translation appears to be unaffected from people 20 to 70 years of age. Thus, as compared with younger people, older people - at least those under 70 years of age - do not appear to have a higher risk of ligamentous knee injuries associated with hamstring dysfunction. Further studies should be conducted to assess whether the present findings also apply to dynamic real-world situations. This would improve the understanding of the relationship between knee stability and ageing. PMID- 18425530 TI - Adaptation of pedaling rate of professional cyclist in mountain passes. AB - The aim of this study was to analyze the pedaling rate (PR) adopted by professional cyclists in different mountain passes. PR, heart rate (HR), velocity and power to overcome gravity were monitored during special (HM), 1st (M1), 2nd (M2) and 3rd (M3) category mountain passes. HM and M1 within high-mountain stages were classified into mountain passes before the final mountain pass of the stage (M-BF) and mountain passes placed in the final of the stage (M-F). PR was significantly higher (P < 0.05) in M3 (82 +/- 1 rpm) than that in M2 (75 +/- 3 rpm), M1 (75 +/- 2 rpm) and HM (73 +/- 1 rpm). Velocity and power output decreased in the following order: M3, M2, M1 and HM. Also, greater values (P < 0.05) were observed in M-BF (24.1 +/- 0.8 km h(-1) and 308.5 +/- 10.4 W) and in M F (17.6 +/- 0.9 km h(-1) and 270.1 +/- 9.9 W). In addition, PR was higher (P < 0.05) in M-BF (79 +/- 2 rpm) than that in M-F (73 +/- 1 rpm). In conclusion, PR was modified according to the characteristics and the race strategies adopted by the cyclists, thus the cyclists chose higher PR to improve their performance. PMID- 18425531 TI - Transient increase in femoral arterial blood flow to the contralateral non exercising limb during one-legged exercise. AB - We studied the effect of exercise intensity and duration on blood flow to the non exercising leg during one-legged dynamic knee extension. Femoral arterial blood flow (FBF) to the non-exercising leg, blood pressure (BP), and heart rate (HR) were monitored during one-legged dynamic knee extension exercise at 15, 30, and 45% maximal voluntary contraction (MVC) in seven healthy females. There was an interaction between exercise intensity and duration for FBF and FVC (P < 0.01). During the initial phase of contralateral leg exercise at all intensities, FBF and femoral vascular conductance (FVC) of non-exercising leg increased, and the increase was larger at higher intensities (P < 0.01). After initial vasodilatation, FBF and FVC decreased to baseline, which suggests the vasoconstriction. However, FBF and FVC gradually increased during exercise at 15% MVC. We conclude that transient vasodilatation at the onset of exercise is followed by gradual change to vasoconstriction in non-exercising limb during dynamic one-legged exercise and these changes are exercise intensity- and duration-dependent. PMID- 18425533 TI - Flow modulates centriole movements in tubular epithelial cells. AB - Kidney cysts are characterized by an abnormal tubular geometry that may result from loss of orientation and random cell divisions during renal development. Since cystic kidney disease is caused by mutations of ciliary proteins and cilia act as flow sensors in the kidney, we examined three polarized events in Madin Darby Canine Kidney cells under flow: cell division, cell migration, and centriole movement. We found that the mitotic orientation of dividing cells was not affected by flow and was randomly distributed in relation to the direction of the flow. Flow did not alter the direction and speed of cell migration in a wound healing assay. However, flow resulted in increased motility of centrioles and biased centrioles to move along the axis of the flow. This effect was lost after flow-induced calcium signaling was abolished by a mutant polycystin 2. Our findings suggest that the cilium may translate fluid flow into altered centriole movements to provide tubular epithelial cells with the spatial orientation required to establish and/or maintain a normal tubular geometry. PMID- 18425532 TI - Four-mode gating model of fast inactivation of sodium channel Nav1.2a. AB - Basic principles of the gating mechanisms of neuronal sodium channels, especially the fast inactivation process, were revealed by a quantitative analysis of the effects of the chemically irreversible modifying agent chloramine T. The compound is known to enhance the open probability of sodium channels by interfering with the inactivation process. The key for the deduction of structure-function relationships was obtained from the analysis of single-channel patch-clamp data, especially the finding that chloramine T-induced modification of inactivation occurred in four steps. These steps were termed modes 1-4 (four-mode gating model), and their temporal sequence was always the same. The kinetic analysis of single-channel traces with an improved two-dimensional dwell-time fit revealed the possible mechanism related to each mode. Similarities to the kinetics of the sodium channel mutant F1489Q led to the assignment of modes 1 and 2 to transient defects in the locking of the inactivation particle (hinged lid). In the third mode, the hinged lid was unable to lock permanently. Finally, in mode 4, the apparent single-channel current was reduced, which could be explained by fast gating, presumably related to the selectivity filter. PMID- 18425534 TI - Cu/Zn superoxide dismutases in developing cotton fibers: evidence for an extracellular form. AB - Hydrogen peroxide and other reactive oxygen species are important signaling molecules in diverse physiological processes. Previously, we discovered superoxide dismutase (SOD) activity in extracellular protein preparations from fiber-bearing cotton (Gossypium hirsutum L.) seeds. We show here, based on immunoreactivity, that the enzyme is a Cu/Zn-SOD (CSD). Immunogold localization shows that CSD localizes to secondary cell walls of developing cotton fibers. Five cotton CSD cDNAs were cloned from cotton fiber and classified into three subfamilies (Group 1: GhCSD1; Group 2: GhCSD2a and GhCSD2b; Group 3: GhCSD3 and GhCSD3s). Members of Group 1 and 2 are expressed throughout fiber development, but predominant during the elongation stage. Group 3 CSDs are also expressed throughout fiber development, but transiently increase in abundance at the transition period between cell elongation and secondary cell wall synthesis. Each of the three GhCSDs also has distinct patterns of expression in tissues other than fiber. Overexpression of cotton CSDs fused to green fluorescent protein in transgenic Arabidopsis demonstrated that GhCSD1 localizes to the cytosol, GhCSD2a localizes to plastids, and GhCSD3 is translocated to the cell wall. Subcellular fractionation of proteins from transgenic Arabidopsis seedlings confirmed that only c-myc epitope-tagged GhCSD3 co-purifies with cell wall proteins. Extracellular CSDs have been suggested to be involved in lignin formation in secondary cell walls of other plants. Since cotton fibers are not lignified, we suggest that extracellular CSDs may be involved in other plant cell wall growth and development processes. PMID- 18425535 TI - Intermitochondrial cement (nuage) in a spermatocytic seminoma: comparison with classical seminoma and normal testis. AB - The nuage, an ultrastructural marker of normal human germ cells (spermatogonia type A and primary spermatocytes), may be found associated with mitochondria (intermitochondrial cement) and/or free in the cytoplasm. Eight specimens from germ cell-related tumours were reviewed to assess whether the nuage could have diagnostic significance in testicular neoplasms. The nuage of neoplastic cells from seven classical seminomas and one spermatocytic seminoma was compared with that from two normal testes. The ultrastructural study demonstrated that only spermatocytic seminoma cells contained both types of nuage and that significantly fewer spermatocytic seminoma cells (28%) contained intermitochondrial cement compared with control spermatogonia type A (81.1%) and primary spermatocytes (47.6%). The data indicate that (1) the detection of the nuage confirms that the phenotype of spermatocytic seminoma is more differentiated than that of classical seminoma; (2) the intermitochondrial cement is an additional example of how a distinctive organelle of a normal cell is preserved in its neoplastic counterpart and (3) if the intermitochondrial cement were found in other cases of spermatocytic seminoma, this organelle of the normal germ cell lineage could be considered as a new ultrastructural marker of the neoplasm. PMID- 18425536 TI - The case of the purple colon. AB - Purple discoloration of the large bowel at autopsy was related to beetroot ingestion and post-mortem changes. PMID- 18425537 TI - Efficacy of plasma therapy in atypical hemolytic uremic syndrome with complement factor H mutations. AB - Atypical hemolytic uremic syndrome (aHUS) frequently results in end-stage renal failure and can be lethal. Several studies have established an association between quantitative or qualitative abnormalities in complement factor H and aHUS. Although plasma infusion and exchange are often advocated, guidelines have yet to be established. Long-term outcome for patients under treatment is still unknown. We describe a patient who, at 7 months of age, presented with aHUS associated with combined de novo complement factor H mutations (S1191L and V1197A) on the same allele. Laboratory investigations showed normal levels of complements C4, C3 and factor H. Plasma exchanges and large-dose infusion therapy resulted in a resolution of hemolysis and recovery of renal function. Three recurrences were successfully treated by intensification of the plasma infusion treatment to intervals of 2 or 3 days. This patient showed good response to large doses of plasma infusions and her condition remained stable for 30 months with weekly plasma infusions (30 ml/kg). Long-term tolerance and efficacy of such intensive plasma therapy are still unknown. Reported secondary failure of plasma therapy in factor H deficiency warrants the search for alternative therapeutic approaches. PMID- 18425538 TI - Pre-emptive infiltration of levobupivacaine is superior to at-closure administration in lumbar laminectomy patients. AB - This is a prospective, randomized, controlled trial that compared the efficacy of different protocols of local tissue infiltration with levobupivacaine or levobupivacaine-methylprednisolone at the surgical site for pain relief after lumbar discectomy. The objective of the study was to determine the efficacy of preemptive wound infiltration with levobupivacaine and levobupivacaine methylprednisolone at the surgical site for pain relief. Patients usually suffer significant pain after lumbar discectomy. Wound infiltration with local anesthetics with or without corticosteroids is one method to address this. A total of 100 patients were randomly allocated to five equal groups as follows: Group I had the musculus multifidi near the operated level infiltrated with 30 mL 0.25% levobupivacaine and 40 mg methylprednisolone just before wound closure; Group II had the same region infiltrated with 30 mL 0.25% levobupivacaine alone before closure; Group III had this region infiltrated with 30 mL 0.25% levobupivacaine and 40 mg methylprednisolone before the incision was made; in Group IV this region was infiltrated with 30 mL 0.25% levobupivacaine alone before incision; and in Group C (controls) this region was infiltrated with 30 mL 0.9% NaCl just before wound closure. Demographics, vital signs, postoperative pain scores and morphine usage were recorded. All four treatment groups showed significantly better results than the control group for most parameters. The treated groups had lower parenteral opioid requirements after surgery, lower incidences of nausea and shorter hospital stays. Further, the data indicate that, compared with infiltration of these drugs at wound closure, preemptive injection of levobupivacaine or levobupivacaine-methylprednisolone into the muscle near the operative site provides more effective analgesia after lumbar discectomy. Our data suggest that preemptive infiltration of the wound site with levobupivacaine alone or combined with methylprednisolone provides effective pain control with reduced opiate dose after unilateral lumbar discectomy. PMID- 18425539 TI - Radiculopathy at the C5/6 intervertebral foramen resulting in isolated atrophy of the deltoid: an aberrant innervation complicating diagnosis. Report of two cases. AB - We present two cases in which the diagnosis was complicated by the presence of a weak muscle innervated by a compressed motor root in the intervertebral foramen (IVF) at an atypical level. The patients were 59- and 53-year-old men; they presented with marked atrophy and weakness predominantly in a unilateral deltoid. Neuroimaging revealed narrowing of the nerve root sleeve at the C5/6 IVF due to a herniated disk or osteophyte. Predominant atrophy and weakness of the deltoid were not consistent with radiculopathy at the C5/6 IVF, i.e. C6 radiculopathy. During an extended observation period to rule out motor neuron disease, their weakness did not spread and the patients underwent posterior medial facetectomy and anterior foraminotomy. This produced marked improvement of the deltoid weakness soon after the operation. We considered unexpectedly wide motor innervation of the C6 nerve root predominantly in the deltoid, anatomic variations in the C5 root such as the trunk of the C5 root entering into the C5/6 IVF, and descending anastomoses connecting the C5 and C6 rootlets as possible explanations. Awareness of this rare presentation may aid in the diagnosis and surgical management of these patients. PMID- 18425540 TI - Mapping iron binding sites on human frataxin: implications for cluster assembly on the ISU Fe-S cluster scaffold protein. AB - Frataxin is an iron binding mitochondrial matrix protein that has been shown to mediate iron delivery during iron-sulfur cluster and heme biosynthesis. There is a high degree of structural homology for frataxin proteins from diverse sources, and all possess an anionic surface defined by acidic residues. In the human protein these residues principally lie on a surface defined by the alpha1 helix and beta1 sheet and the impact of multiple substitutions of these carboxylate residues on iron binding is described. Full-length human frataxin has previously been shown to undergo self-cleavage to produce a truncated form both in vitro and in vivo. This truncated protein has been shown to bind approximately seven iron centers that are presumably associated with the acidic patch. Relative to this native protein, the stoichiometry decreases according to the number and sites of mutations. Nevertheless, the iron-dependent binding affinity of each frataxin derivative to the iron-sulfur cluster scaffold protein ISU is found to be similar to that of native frataxin, as defined by isothermal titration calorimetry experiments, requiring only one iron center to promote nanomolar binding. While frataxins from various cell types appear to bind differing numbers of iron centers, the physiologically relevant number of bound irons appears to be small, with significantly higher binding affinity following complex formation with partner proteins (micromolar compared with nanomolar binding). By contrast, in reconstitution assays for frataxin-promoted [2Fe-2S](2+) cluster assembly on ISU, one derivative does display a modestly lower reconstitution rate. The overall consensus from these data is to consider a pool of potential sites that can stably bind an iron center when bridged to a variety of physiological targets. PMID- 18425541 TI - SAR comparative studies on pyrimido[4,5-b][1,4] benzothiazine derivatives as 15 lipoxygenase inhibitors, using ab initio calculations. AB - The enzyme inhibitory activity of a new group of 2-substituted pyrimido[4,5 b][1,4]benzothiazines on soybean 15-lipoxygenase (15-LO) was evaluated and compared with those of their 4-methyl analogs using ab initio calculations. The results of these studies showed that the lack of 4-methyl substituent in the pyrimido[4,5-b][1,4] benzothiazine molecules greatly reduces their 15-LO inhibitory activities. PMID- 18425542 TI - Modification by glucose of the flocculent phenotype of a Kloeckera apiculata wine strain. AB - We have evaluated the induction of the flocculent phenotype of Kloeckera apiculata by glucose mc1 and propose a pathway involved in carbohydrate flocculation induction. Pulses of glucose were given to cells growing in glucose poor medium (2 g l(-1)) and the flocculation percentage was measured. To elucidate the mechanism involved in flocculation induction, cycloheximide was injected into the cultures 120 min before the glucose pulse. 2,4-Dinitrophenol or cAMP was added to the media instead, or simultaneously with glucose, while a protein kinase A (PKA) inhibitor was added 30 min before the glucose pulse. With 20 and 50 g l(-1) glucose pulse, the yeast flocculation percentage arises to 55 and 65%, respectively. The quantity of proteins and the reflocculating capacity of a lectinic protein extract from the yeast cell wall increase as the concentration of glucose pulse was higher. Cycloheximide prevented the glucose induced flocculation, while cAMP or 2,4-dinitrophenol increased it 4- and 5-fold, respectively. PKA inhibitor completely prevented the glucose induction flocculation. The flocculent phenotype of K. apiculata mc1 was induced by glucose and the mechanism seems to imply de novo protein (lectin) synthesis via the PKA transduction pathway. This work contributes to the elucidation of the mechanism involved in flocculation induction by glucose of a non-Saccharomyces wine yeast, K. apiculata, which has not been reported. The induction of flocculation by glucose could be a biotechnological tool for the early removal of the indigenous microorganisms from the grape must before the inoculation of a selected starter strain to conduct the alcohol fermentation. PMID- 18425543 TI - Effects of functional electrical stimulation in denervated thigh muscles of paraplegic patients mapped with T2 imaging. AB - OBJECT: Functional electrical stimulation (FES) for paraplegic patients, with the long-term goal of ultimately restoring muscle function, is associated with several positive effects: improvement of blood circulation, skin condition, peripheral trophism and metabolism, prophylaxis against decubitus ulcer and better physical fitness. Since fibres of denervated muscles (lacking a supplying nerve) need to be activated directly, the fraction of elicited muscle tissue follows the geometric distribution of the electrical field, which can be simulated using electrophysiological computer models. Experimental validation of these results, however, has not yet been established. MATERIALS AND METHODS: We acquired T (2) parameter images using a multislice multi-spin-echo MR sequence before and immediately after FES in nine denervated paraplegic patients and three healthy subjects in order to visualise the geometric distribution of activation by electrically induced muscle stimulation in denervated versus innervated (healthy) thigh muscle. RESULTS AND CONCLUSION: After realigning and normalisation, maps of relative T (2) increase were calculated. The results demonstrate that the spatial distribution of short-term effects of FES of denervated muscle tissue of paraplegic patients who regularly perform FES can be visualised by T (2) parameter images. This may be used to refine models of the electrical field of FES in muscle and fibre activation in the future. PMID- 18425544 TI - Pancreatic duct evaluation: accuracy of portal venous phase 64 MDCT. AB - BACKGROUND: Many patients presenting with nonspecific signs and symptoms often receive CT scans using general protocols, not optimized to evaluate for pancreatic pathology. Therefore the purpose of this study was to evaluate portal venous phase 64 multi-row detector CT (MDCT) scans for detecting pancreatic duct strictures, stones, pancreas divisum, and communication between pancreatic ducts and cystic pancreatic lesions. METHODS: Institutional review board approval with waived informed consent was obtained for this HIPAA-compliant study. We included all patients that underwent abdominal, portal venous phase, intravenous contrast enhanced 64 MDCT scans between 6/7/05 and 5/01/07 and MR cholangiopancreatography (MRCP) or endoscopic retrograde pancreatography (ERCP) within 2 months of the CT. This yielded 93 patients (42 males, 51 females) with a mean age of 59 years. In addition to CT, 75 patients underwent MRCP and 37 patients underwent ERCP. Two radiologists independently evaluated the CT images, including multiplanar and minimum intensity pixel projection reformations, for pancreatic duct strictures, stones, pancreas divisum, or cystic pancreatic lesions. The latter were classified as communicating or not communicating with the pancreatic ducts. Findings on ERCP or MRCP were used to calculate diagnostic performance parameters. RESULTS: On standard of reference examinations, 15 (16%) of the 93 patients had a pancreatic duct stricture. The sensitivity and the specificity for Observer 1 were 87% and 100%, respectively; for Observer 2, 100% and 100%, respectively. Six (6%) of the 93 patients had main pancreatic duct stones. The sensitivity and the specificity for Observer 1 were 83% and 100%, respectively; for Observer 2, 100% and 99%, respectively. Five (5%) patients had pancreas divisum; Observer 1 correctly identified four and Observer 2 correctly identified three cases. Eleven (12%) patients had cystic pancreatic lesions. Observer 1 correctly determined whether or not there was communication between the cystic pancreatic lesion and the pancreatic duct in ten cases; Observer 2 correctly made this determination in nine cases. CONCLUSION: Portal venous phase 64 MDCT images are moderately sensitive and highly specific for detecting pancreatic duct stricture, stones, and pancreas divisum and moderately accurate for detecting communication between pancreatic ducts and cystic pancreatic lesions. PMID- 18425545 TI - Target molecular therapies: methods to enhance and monitor tumor drug delivery. AB - Noninvasive monitoring/quantification of drug delivery to tumors is an ideal goal of chemotherapy treatment. However, the ability to overcome the barriers to the developing of targeted therapies-along with the physiological barriers that the tumor presents-is still needed. Recent advances demonstrate that targeted therapies can be used for diagnostic and therapeutic applications. The most mature of these technologies are liposomes that encapsulate a therapeutic drug in conjunction with a contrast agent. Through selective manipulation of the liposome composition, modification of the tumor microphysiology, and temporal sequencing of liposome administration with tumor microphysiology modification improvement in efficacy can be achieved. The future application of these targeted therapies will allow the radiologist to become a more central member of the cancer treatment team, further expanding the field and the radiologist's unique skills. PMID- 18425546 TI - Evaluation of acute mesenteric ischemia: accuracy of biphasic mesenteric multi detector CT angiography. AB - The purpose of this study was to explore the accuracy of multi-detector row helical CT (MDCT), using a biphasic mesenteric angiography protocol for evaluation of acute mesenteric ischemia (AMI). In total, 79 consecutive patients with clinical signs of AMI underwent contrast enhanced 16- or 40-channel MDCT. MDCT findings were correlated with surgery, endoscopy and clinical outcome. Sensitivity, specificity, and positive and negative predictive values were calculated using the patients in which AMI had been excluded as a control group. In 28 patients the final diagnosis was AMI. In 27 patients (96.4%) MDCT correctly diagnosed AMI (specificity of 97.9%). A sensitivity of 93%, specificity of 100%, and positive and negative predictive values of 100% and 94%, respectively were achieved for the CT findings of visceral artery occlusion, intestinal pneumatosis, portomesenteric venous gas or bowel wall thickening in combination with either portomesenteric thrombosis or solid organ infarction. Our findings suggest that mesenteric MDCT angiography is an accurate tool for fast diagnostic work-up of patients with suspected AMI. PMID- 18425547 TI - Key performance indicators in British military trauma. AB - BACKGROUND: Key performance indicators (KPI) are tools for assessing process and outcome in systems of health care provision and are an essential component in performance improvement. Although KPI have been used in British military trauma for 10 years, they remain poorly defined and are derived from civilian metrics that do not adjust for the realities of field trauma care. Our aim was to modify current trauma KPI to ensure they more faithfully reflect both the military setting and contemporary evidence in order to both aid accurate calibration of the performance of the British Defence Medical Services and act as a driver for performance improvement. METHOD: A workshop was convened that was attended by senior, experienced doctors and nurses from all disciplines of trauma care in the British military. "Speciality-specific" KPI were developed by interest groups using evidence-based data where available and collective experience where this was lacking. In a final discussion these were streamlined into 60 KPI covering each phase of trauma management. CONCLUSION: The introduction of these KPI sets a number of important benchmarks by which British military trauma can be measured. As part of a performance improvement programme, these will allow closer monitoring of our performance and assist efforts to develop, train, and resource British military trauma providers. PMID- 18425548 TI - Adrenal incidentalomas: experience in a developing country. AB - BACKGROUND: The incidence of adrenal incidentalomas is rising worldwide. There is a concern about malignancy in large incidentalomas. There are few published articles from developing countries on this entity. The aim of this study was to analyze the clinical presentation, functional status, and final diagnosis of adrenal incidentalomas and, in particular, to look into the incidence of adrenal cortical carcinoma (ACC) in large adrenal incidentalomas managed at a tertiary referral hospital in northern India. METHODS: This is a retrospective study (January 1991-December 2005) of 59 patients with adrenal incidentaloma managed at our department. The mean tumor diameter was 7.8 +/- 4.0 cm. In all cases, clinical details, radiology findings, laboratory findings, intervention details, histology findings, and follow-up data were noted. Six patients in which the mass was ultimately found to arise from extra-adrenal tissue were excluded from final analysis RESULTS: Mean age of the patients was 46 +/- 12 years (M:F = 1:1.1). The incidentaloma measured more than 3 cm in 91% of cases and more than 6 cm in 70% of cases. The clinical scenarios leading to detection of incidentaloma included abdominal (56.6%), genitourinary (24.5%), and systemic complaints (15.1%) and routine medical checkup (3.7%). Most of cases were found by ultrasonography (n = 41), and remaining by CT scan (n = 10), and MRI (n = 2). The incidence of functioning tumors was 41.5% (hypercatecholinism 37.7 % and hypercortisolism 1.9%). Forty-nine patients were operated on. The important final pathology included ACC (7.5%), pheochromocytoma (PCC) (43%), adrenal cysts (13.2%), myelolipoma (11.3%), and inflammatory lesions (9.4%). CONCLUSION: In our experience, the incidence of PCC was high among large adrenal incidentalomas while that of ACC was lower than expected. Inflammatory lesions contribute to a significant number of cases of incidentaloma in developing countries. Adrenalectomy is a justified procedure for large incidentalomas, although the concern may not always be an underlying malignancy. PMID- 18425550 TI - INDIAM--an e-learning system for the interpretation of mammograms. AB - We propose the design of a teaching system named Interpretation and Diagnosis of Mammograms (INDIAM) for training students in the interpretation of mammograms and diagnosis of breast cancer. The proposed system integrates an illustrated tutorial on radiology of the breast, that is, mammography, which uses education techniques to guide the user (doctors, students, or researchers) through various concepts related to the diagnosis of breast cancer. The user can obtain informative text about specific subjects, access a library of bibliographic references, and retrieve cases from a mammographic database that are similar to a query case on hand. The information of each case stored in the mammographic database includes the radiological findings, the clinical history, the lifestyle of the patient, and complementary exams. The breast cancer tutorial is linked to a module that simulates the analysis and diagnosis of a mammogram. The tutorial incorporates tools for helping the user to evaluate his or her knowledge about a specific subject by using the education system or by simulating a diagnosis with appropriate feedback in case of error. The system also makes available digital image processing tools that allow the user to draw the contour of a lesion, the contour of the breast, or identify a cluster of calcifications in a given mammogram. The contours provided by the user are submitted to the system for evaluation. The teaching system is integrated with AMDI-An Indexed Atlas of Digital Mammograms-that includes case studies, e-learning, and research systems. All the resources are accessible via the Web. PMID- 18425549 TI - Cloning and use of a coral 36B4 gene to study the differential expression of coral genes between light and dark conditions. AB - This paper aims to validate reference genes for gene expression studies between light and dark conditions in the scleractinian coral Stylophora pistillata for future gene expression studies of the "light-enhanced calcification" phenomenon. For this purpose, we cloned, sequenced, and characterized a candidate reference gene, the 36B4 gene from the coral S. pistillata, and validated 36B4 and beta actin as reference genes. To illustrate the future applications of these reference genes, we tested the dark and light expression of two photosynthetic genes (Rubisco and D1 protein of the photosystem II) and two genes encoding proteins involved in calcium transport for coral calcification (a calcium ATPase and a calcium channel). Results show that both photosynthetic genes are enhanced during the light when standardized against 36B4 and beta-actin, whereas the two genes encoding proteins involved in calcium transport are not differentially expressed between light and dark conditions. The characterization of a coral 36B4 and the establishment of such valid reference genes will be useful for future gene expression studies between diverse conditions (aposymbiotic/symbiotic, stress/control, light/dark conditions) in scleractinian corals. PMID- 18425551 TI - Are you worried about bowel accidents? Quality of life in fecal incontinence. PMID- 18425552 TI - Biological behavior of perihepatic lymph node metastasis and its impact on prognosis following liver resection for colorectal metastases. PMID- 18425553 TI - Beyond the ivory tower: outcomes following surgery for ampullary cancer. PMID- 18425554 TI - Cancer clinical trials accrual: missed opportunities to address disparities and missed opportunities to improve outcomes for all. PMID- 18425555 TI - Lesions of "uncertain malignant potential" diagnosed by vacuum-assisted breast biopsy: an unclear management? PMID- 18425556 TI - Assessment of cerebral glucose metabolism in cat deafness model: strategies for improving the voxel-based statistical analysis for animal PET studies. AB - PURPOSE: The aim of this study was to establish the procedures for 3D voxel-based statistical analysis of 2-deoxy-2-[(18)F]fluoro-D-glucose-positron emission tomography (FDG-PET) images of a cat's brain obtained using a small animal dedicated PET system and to assess the utility of this approach in investigating the cerebral glucose metabolism in an animal model of cortical deafness. PROCEDURES: This study compared several different strategies for the spatial processing of PET data acquired twice from eight cats before and after inducing deafness in terms of the comparability of the statistical analysis results to the established pattern of the cerebral glucose metabolic changes in the deaf animals. RESULTS: The accuracy of the spatial preprocessing procedures and the statistical significance of the comparison were improved by removing the background activities outside the brain regions. The use of the spatial normalization parameters obtained from the mean image of the realigned data set for individual data also helped improve the statistical significance of the paired t testing. It was also found that an adjustment of the registration options was also important for increasing the precision of the realignment. CONCLUSIONS: A method for voxel-based analysis of the PET data of a cat's brain was optimized. The results demonstrated the high localization accuracy and specificity of this method, which is expected to be useful for examining the brain PET data of medium-sized animals such as cats. PMID- 18425557 TI - Histological alterations following bariatric surgery: pilot study. AB - BACKGROUND: Macroscopic clinical evidence of tissue damaging following bariatric surgery pushed plastic surgeons to presume microscopic alterations as well. METHODS: Five preliminary cases studied randomly, and compared with healthy tissues, confirmed these initial suspects. A deeper and wider study has then been structured. RESULTS: Preliminary results are so evident to encourage us to carry on an estimated 2-3 years multidisciplinary study. CONCLUSIONS: What we want to study is if histological post-bariatric alterations are confirmed, and if these can be considered responsible for higher complication rate in body contouring following bariatric surgery. PMID- 18425558 TI - The reamer/irrigator/aspirator reduces femoral canal pressure in simulated TKA. AB - Inserting the femoral intramedullary alignment rod during total knee arthroplasty (TKA) can generate high intramedullary pressure, which increases the risk of intraoperative complications caused by fat embolism. Despite modifications to the surgical procedure, the best method to prevent this increase in pressure remains unknown. The reamer/irrigator/aspirator is a surgical instrument designed for use during femoral canal entry to increase the canal size and remove intramedullary fat and may prevent this pressure increase. We posed two hypotheses: (1) using the reamer/irrigator/aspirator system will result in lower maximum femoral intramedullary pressure than using only conventional instrumentation during the initial steps of a TKA; and (2) using the reamer/irrigator/aspirator system in the initial steps of TKA will result in a mean maximum intramedullary pressure less than 200 mm Hg. We simulated a TKA on 14 cadaveric femurs to compare the femoral intramedullary pressure using both methods. Considerable decreases in femoral intramedullary pressure of 86% proximally and 87% distally were obtained by using the reamer/irrigator/aspirator system. The mean maximum pressure using the reamer/irrigator/aspirator system was less than 200 mm Hg. Additional clinical studies are needed to confirm any reduction in complications using the reamer/irrigator/aspirator system. PMID- 18425560 TI - A comparison of intramedullary and juxtacortical low-grade osteogenic sarcoma. AB - While low-grade juxtacortical and low-grade intramedullary osteogenic sarcomas are histologically indistinguishable, they have been studied as separate entities. We retrospectively reviewed the clinical, radiographic, histologic features and treatment of 59 patients treated surgically to compare the rate of local recurrence, grade progression, and survival between low-grade intramedullary and low-grade juxtacortical osteogenic sarcoma. Forty-five (76%) patients were treated for low-grade juxtacortical osteogenic sarcoma and 14 (24%) were treated for low-grade intramedullary osteogenic sarcoma. Local recurrence rates of 7% were similar for both groups studied. The rate of distant metastases was also similar for both groups. . The rate of dedifferentiation for the entire group was 29%. Dedifferentiated lesions were treated with adjuvant chemotherapy in 16 of 17 cases. Recurrence preceded dedifferentiation in four cases. Five-year survival was over 90% in both groups. Low-grade intramedullary and low-grade juxtacortical osteogenic sarcoma were clinically indistinguishable with identical rates of local recurrence, distant metastases, dedifferentiation, and survival. LEVEL OF EVIDENCE: Level III, therapeutic study. See the Guidelines for Authors for a complete description of levels of evidence. PMID- 18425559 TI - Collagens, proteoglycans, MMP-2, MMP-9 and TIMPs in human achilles tendon rupture. AB - Tendon integrity depends on the extracellular matrix (ECM) metabolism which is regulated by proteolytic enzymes. However, it is unclear which enzymes play a role in tendon rupture. We studied the ECM of 19 ruptured human Achilles tendons, comparing the composition of specimens harvested close to the rupture with specimens harvested from an apparently healthy area in the same tendon. We compared gene expression of collagen Type I, decorin, and versican including enzymes involved in their metabolism as matrix metalloproteases (MMP-2 and -9) and tissue inhibitory of metalloproteinase (TIMP-1 and -2) using real-time PCR, zymography and FACE analysis. We found greater gene expression of proteoglycan core protein decorin and versican, collagen Type I, MMPs and TIMPs in the tendon rupture. Zymography analysis, reflecting expression of enzymatic activity, confirmed the gene expression data at protein level. Carbohydrate content was greater in the macroscopically healthy area than in the ruptured area. In the ruptured area, we found increased core protein synthesis but without the normal glycosaminoglycan production. The tissue in the area of rupture undergoes marked rearrangement at molecular levels and supports the role of MMPs in the pathology. PMID- 18425561 TI - Protecting a patellar ligament reconstruction after proximal tibial resection: a simplified approach. AB - Limb salvage in tumor surgery has encouraged the development of megaprostheses. However, reattaching the ligamentum patellae poses a particular problem: avulsion and/or extensor lag may lead to poor function. We describe a new technique of patellar ligament reconstruction. The technique involves reattachment of the patellar ligament to the tibial tuberosity of the proximal tibial megaprosthesis, which has a porous surface created, and the repair is protected with a cerclage wire through the patella and the prosthesis. In 10 consecutive patients, the range of motion averaged 95 degrees (median, 90 degrees ; range, 70 degrees -120 degrees ), and the mean extension lag averaged 4 degrees (median, 0 degrees ; range, 0 degrees -20 degrees ). We had one case of patellar ligament avulsion. This technique resulted in good quadriceps function and a low incidence of complications. LEVEL OF EVIDENCE: Level IV, therapeutic study. PMID- 18425562 TI - Analysis of the pathology section of the OITE will aid in trainee preparation. AB - We analyzed the tumor-related questions on the Orthopaedic In-Training Examinations (OITE) over a 5-year period (2002-2006). Each year approximately 27 of the 275 examination questions (10%) are tumor-related. Malignant conditions are tested more than benign ones (1.2:1). Approximately nine questions per year are immediate recall of specific entities, while eight questions tested the examinees' ability to establish a diagnosis from imaging studies with or without biopsy material. Participants were required to establish a diagnosis from imaging studies and/or biopsy material and then choose treatment an average of seven times per examination. The examinees' abilities in evaluation and decision making of patients was only tested an average of twice during each examination. Analysis of the content and type of question on the OITE might enable trainees to prepare more systematically. We determined the content, classified the questions, and quantified the interpretive material. PMID- 18425564 TI - A case of granuloma in the occipital lobe of a patient with Wegener's granulomatosis. AB - We report a case of Wegener's granulomatosis (WG) with central nervous system (CNS) involvement in a woman who complained of bilateral visual disturbance. The intracranial necrotizing granulomatous lesion was confirmed by MR imaging and brain biopsy. After high-dose oral corticosteroid treatment, not only clinical manifestations but also laboratory tests improved. Moreover, the CNS lesion completely regressed. This suggests that high-dose corticosteroid alone might be effective in treating WG with CNS involvement. PMID- 18425565 TI - Osteoclast cell fusion: mechanisms and molecules. AB - Osteoclasts are bone-resorbing multinuclear polykaryon that are essential for bone remodeling and are formed through cell fusion of mononuclear macrophage/monocyte-lineage hematopoietic precursors. In arthritic joints, a large number of activated osteoclasts can be detected, which are suggested to be causative of bone erosion in rheumatoid arthritis. It has been fully established that osteoclastogenesis is critically regulated by several key essential factors, such as M-CSF and RANKL. However, regarding their most characteristic property, i.e., cell fusion to form giant polykaryons, there are still miscellaneous questions to be clarified, although several molecules have been shown to be critically involved in this process. Here we review the latest knowledge about osteoclastogenic cell fusion and novel concepts underlying the characteristic phenomenon. Because cell fusion is a genuine property of mature osteoclasts, modulating this process will become a promising therapeutic tool for bone resorptive disorders in the future. PMID- 18425563 TI - Water flow on erbium:yttrium-aluminum-garnet laser irradiation: effects on dental tissues. AB - Since lasers were introduced in dentistry, there has been considerable advancement in technology. Several wavelengths have been investigated as substitutes for high-speed air turbine. Owing to its high absorbability in water and hydroxyapatite, the erbium:yttrium-aluminum-garnet (Er:YAG) laser has been of great interest among dental practitioners and scientists. In spite of its great potential for hard tissue ablation, Er:YAG laser effectiveness and safety is directly related to an adequate setting of the working patterns. It is assumed that the ablation rate is influenced by certain conditions, such as water content of the target tissue, and laser parameters. It has been shown that Er:YAG irradiation with water coolant attenuates temperature rise and, hence, minimizes the risk of thermally induced pulp injury. It also increases ablation efficiency and enhances adhesion to the lased dental tissue. The aim of this review was to obtain insights into the ablation process and to discuss the effects of water flow on dental tissue ablation using Er:YAG laser. PMID- 18425566 TI - Optic disc hemorrhages in a patient with hemophilia A. AB - To report a hemophilia patient complicated with optic disc hemorrhages. A 13-year old boy presented to our emergency room with a black shadow in the left eye for 1 day. The best-corrected visual acuity was 6/6 in both eyes. Peripapillary retinal and subretinal hemorrhages were found in the left eye. Result of the laboratory examination showed an extremely low level of coagulation factor VIII (1.9%). Factor VIII concentrate was given for 8 weeks. A follow-up 3 months later showed absorption of the hemorrhages, black shadow diminished, and the vision was 6/6. PMID- 18425567 TI - Tryptophan administration induces oxidative stress in brain cortex of rats. AB - Despite the significant brain abnormalities, the neurotoxic mechanisms of brain injury in hypertryptophanemia are virtually unknown. In this work, we determined the thiobarbituric acid-reactive substances, 2',7'-dihydrodichlorofluorescein oxidation, reduced glutathione and the activities of catalase, superoxide dismutase and glutathione peroxidase in cerebral cortex from rats loaded with L tryptophan. High L-tryptophan concentrations, similar to those found in hypertryptophanemic patients were induced by three subcutaneous injections of saline-buffered tryptophan (2 micromol/g body weight) to 30-day-old Wistar rats. The parameters were assessed 1 h after the last injection. It was observed that tryptophan significantly increased thiobarbituric acid-reactive substances, 2',7' dihydrodichlorofluorescein oxidation and reduced glutathione, whereas it reduced catalase activity. Pre-treatment with taurine (1.6 micromol/g of body weight), or alpha-tocopherol plus ascorbic acid (40 and 100 microg/g body weight, respectively) prevented those effects of tryptophan, reinforcing the hypothesis that tryptophan induces oxidative stress in brain cortex of the rats. Therefore, these findings also occur in human hypertryptophanemia or in other neurodegenerative diseases in which tryptophan accumulates, then oxidative stress may be involved in the mechanisms leading to the brain injury observed in patients affected by these disorders. PMID- 18425568 TI - Failure of acute administration with proteasome inhibitor to provide a model of Parkinson's disease in mice. AB - We investigated to determine whether acute administration of proteasome inhibitor can cause dopaminergic cell loss in mice, in comparison with that of 1-methyl-4 phenyl-1,2,3,6-tetrahydropyridine (MPTP). The four intraperitoneally administrations of MPTP at 1-h intervals to mice decreased significantly the concentration of dopamine, 3,4-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA) in the striatum after 5 days, in comparison with vehicle treated animals. In contrast, the three subcutaneously administrations of carbobenzoxy-L-gamma-t-butyl-L-glutamyl-L-alanyl-L-leucinal (PSI) did not show significant changes in the concentration of dopamine, DOPAC and HVA in the striatum after 5 days, in comparison with vehicle-treated animals. Our Western blot analysis also showed that the four administrations of MPTP at 1-h intervals to mice produced a significant reduction of anti-tyrosine hydroxylase antibody (TH) protein levels in the striatum after 5 days after. In PSI-treated mice. In contrast, no significant change of TH protein levels was observed in the striatum 5 days after the final treatment with PSI. Furthermore, a significant decrease of TH protein levels was observed in the striatum of MPTP-treated mice, as compared with PSI-treated animals. The present study demonstrates that the acute treatment with proteasome inhibitor PSI did not cause the dopaminergic neurotoxicity in mice, as compared with acute treatment with MPTP. Thus, our findings suggest that acute proteasome inhibition is not a reliable model for Parkinson's disease. PMID- 18425569 TI - Advancement in antithrombotics for stroke prevention in atrial fibrillation. AB - The focus of this review is the evolving field of antithrombotic drug therapy for stroke prevention in patients with atrial fibrillation (AF). The current standard of therapy includes warfarin, acenocoumarol and phenprocoumon which have proven efficacy by reducing stroke by 68% against placebo. However, a narrow therapeutic index, wide variation in metabolism, and numerous food and drug interactions have limited their clinical application to only 50% of the indicated population. Newer agents such as direct thrombin inhibitors, factor Xa inhibitors, factor IX inhibitors, tissue factor inhibitors and a novel vitamin K antagonist are being developed to overcome the limitations of current agents. The direct thrombin inhibitor dabigatran is farthest along in development. Further clinical trial testing, and eventual incorporation into clinical practice will depend on safety, efficacy and cost. Development of a novel vitamin K antagonist with better INR control will challenge the newer mechanistic agents in their quest to replace the existing vitamin K antagonists. Till then, the large unfilled gap to replace conventional agents remains open. This review will assess all these agents, and compare their mechanism of action, stage of development and pharmacologic profile. PMID- 18425570 TI - Motor pattern selection by combinatorial code of interneuronal pathways. AB - We use a modeling approach to examine ideas derived from physiological network analyses, pertaining to the switch of a motor control network between two opposite control modes. We studied the femur-tibia joint control system of the insect leg, and its switch between resistance reflex in posture control and "active reaction" in walking, both elicited by the same sensory input. The femur tibia network was modeled by fitting the responses of model neurons to those obtained in animals. The strengths of 16 interneuronal pathways that integrate sensory input were then assigned three different values and varied independently, generating a database of more than 43 million network variants. We demonstrate that the same neural network can produce the two different behaviors, depending on the combinatorial code of interneuronal pathways. That is, a switch between behaviors, such as standing to walking, can be brought about by altering the strengths of selected sensory integration pathways. PMID- 18425571 TI - Genetic characterization of avian influenza viruses isolated from waterfowl in southern part of South Korea in 2006. AB - Aquatic birds are a reservoir of all known influenza A viruses. Avian influenza viruses have played a major role in the creation of pandemic influenza viruses in humans. In this study, we genetically characterized genes of nine isolates from waterfowl in Eulsukdo, a congregating place for migratory birds on the flyway of migration from Siberia, which is located in the southern part of South Korea. Phylogenic analysis showed that HA and NA genes of isolates belonged to Eurasian lineage, and lineage analysis showed that NS, PB1, PA, NP, and M genes of isolates clustered with Eurasian lineage, and PB2 genes of isolates belonged to North American or Eurasian lineage. Results suggest that the interregional transmission of genes of avian influenza viruses may occur in the migratory birds. PMID- 18425572 TI - Characterization of a protein tyrosine phosphatase gene CvBV202 from Cotesia vestalis polydnavirus (CvBV). AB - Cotesia vestalis (Haliday) (Hymenoptera: Braconidae) is an endoparasitoid of the larval stage of the diamondback moth, Plutella xylostella (L.) (Lepidoptera: Plutellidae) and injects a polydnavirus (CvBV) into its host during oviposition. In this paper we characterize CvBV202 and its product. CvBV202 is located on segment S2 of CvBV genome; it has a size of 984 bp and encodes a putative protein of 328 amino acids, including protein phosphatase domain and tyrosine-specific protein phosphatase domain. Gene transcripts were detected in extracts of the host as early as 2 h post-parasitization (p.p.) and continued to be detected for 6 days. Tissue-specific patterns of this gene expression showed that CvBV202 had a close relationship with the host's physiological alternations including immunosuppression, modulation of hormone titer, and nutrition metabolism. The protein was detected in the parasitized hosts at 12 h p.p. using western blot assay. The product of CvBV202 was found to be around 59 kDa, much larger than the predicted molecular weight of 37.8 kDa, suggesting that post-translational modification of CvBV202 occurs in host cells, corresponding with the existence of many post-translational modification sites. Immunofluorescence staining and confocal laser scanning microscopy revealed that CvBV202 and the fused protein eGFP-CvBV202 were observed both in the nuclear region and cytoplasm of the hemocytes of the naturally parasitized host larvae and rBac-eGFP-CvBV202-infected Tn-5B1-4 cells, respectively. PMID- 18425573 TI - Sexual harassment at work in the United States. AB - Using nationally representative data from the 1992 U.S. National Health and Social Life Survey, this study queried the prevalence and risk factors of lifetime workplace sexual harassment among both women and men. Among those aged 18-60 reporting ever having worked, 41% of women (CI, 37-44) reported any workplace harassment over their lifetime, with men's harassment prevalence significantly lower, at 32% (CI, 29-35). In the youngest age groups (those in their 20s or younger), there was no statistically significant difference between women's and men's harassment prevalence. Multivariate analysis of risk factors suggested that, in contrast to much of the harassment literature, among both genders workplace harassment seemed to have at least as much to do with a system of "routine activities" mechanisms-a victim's conscious or unconscious sexual signaling, more exposure to potential harassers, and a perpetrator's lower cost of harassment-as with unobserved differences in power between victim and perpetrator. Strikingly, both women's and men's harassment was strongly linked to markers of sexualization, whether early developmental factors or behavioral patterns in adulthood-a mechanism insufficiently emphasized in the harassment literature. PMID- 18425574 TI - Elevated body mass index (BMI) does not adversely affect in vitro fertilization outcome in young women. AB - OBJECTIVE: To determine if elevated body mass index in young women with normal ovarian reserve was associated with poorer ovarian response, difficulty at embryo transfer, and lower clinical pregnancy rates. MATERIALS AND METHODS: Retrospective study of 417 first, fresh in vitro fertilization cycles performed between October 2004 and December 2006. All women were under the age of 35 and had normal cycle day 3 follicle stimulating hormone and estradiol levels. Subjects were divided into groups by BMI: <18.5, 18.5-24.9, 25-29.9, > or = 30. RESULTS: Cancellation rates, peak estradiol levels, and mean number of oocytes retrieved were similar in all groups. There was a trend toward increasing difficulty in visualizing the air bubble at time of embryo transfer and lower implantation rates at higher body mass indices. Clinical and ongoing pregnancy rates were similar among groups. CONCLUSION: Obesity in young women does not adversely affect clinical pregnancy rates in patients treated with in vitro fertilization. PMID- 18425575 TI - Enhanced 5-HT(2A) receptor status in the hypothalamus and corpus striatum of ethanol-treated rats. AB - AIM: Brain is the major target for the actions of ethanol and it can affect the brain in a variety of ways. In the present study we have investigated the changes in 5-HT level and the 5-HT(2A) receptors in the ethanol-treated rats. METHODS: Wistar adult male rats of 180-200 g body weight were given free access to 15% (v/v) (approx.7.5 g/Kg body wt./day) ethanol for 15 days. Controls were given free access to water for 15 days. Brain 5-HT and its metabolites were assayed by high performance liquid chromatography (HPLC) integrated with an electrochemical detector (ECD) fitted with C-18-CLS-ODS reverse phase column. 5-HT(2A) receptor binding assay was done with different concentrations of [3H] MDL 100907. RESULTS: The hypothalamic 5-HT content significantly increased (P < 0.001) with a decreased (P < 0.001) 5-HIAA/5-HT turnover in the ethanol-treated rats when compared to control. The corpus striatum 5-HT content significantly decreased (P < 0.01) with increased (P < 0.01) 5-HIAA/5- HT turnovers in the ethanol-treated rats when compared to control. Scatchard analysis of [(3)H] MDL 100907 against ketanserin in hypothalamus showed a significant increase (P < 0.001) in B(max )with a decreased affinity (P < 0.001) in ethanol-treated rats when compared to control. The competition curve for [3H] MDL 100907 against ketanserin fitted one site model in all the groups with unity as Hill slope value. An increased K(i) and log (EC(50)) value were also observed in ethanol-treated rats when compared to control. Scatchard analysis of [3H] MDL 100907 against ketanserin in the corpus striatum of ethanol-treated rats showed a significant increase (P < 0.001) in B(max) and in affinity (P < 0.01) when compared to control. The change in affinity of the receptor protein in both corpus striatum and hypothalamus shows an altered receptor. The competition curve for [(3)H] MDL 100907 against ketanserin fitted one-site model in all the groups with unity as Hill slope value. There was no significant change in K(i) and log (EC (50)) value in ethanol treated rats when compared to control. CONCLUSION: The present study demonstrated the enhanced 5-HT(2A) receptor status in hypothalamus and corpus striatum. The ethanol-induced enhanced 5-HT(2A) receptors in the hypothalamus and corpus striatum has clinical significance in the better management of ethanol addiction. This will have therapeutic application. PMID- 18425576 TI - Time-varying pattern of recurrence risk for Chinese breast cancer patients. AB - PURPOSE: To analyze the rule of recurrence risk for breast cancer patients after surgery in order to get characteristics for Chinese breast cancer. METHODS: We performed a retrospective study of 2,213 female unilateral breast cancer patients undergoing surgery in Cancer hospital of Fudan University, Shanghai, China. Survival curves were performed with Kaplan-Meier method and annual recurrence hazard was estimated by hazard function. RESULTS: Annual recurrence hazard curve for entire population showed a double-peaked pattern, with a first major recurrence surge reaching the maximum at the second year after surgery and a second recurrence peak near the 9.5th year. The analysis according to tumor size, axillary lymph node (ALN) status (T1 + T2 versus T3, node-positive versus node negative) and menopausal status as well as hormone receptor (HR) status proved that the double-peaked pattern was present in each subgroup. Compared with ER positive tumors, ER-negative breast cancers were more likely to recur early. As far as Her-2/neu status was concerned, Her-2/neu-positive patients exhibited a relatively pronounced and variable pattern and tended to have more relapse across all time periods (P<0.05). CONCLUSIONS: The double-peaked pattern observed in our patients gives further support to tumor dormancy hypothesis. It contributes to different treatment strategies (e.g. the type and timing of adjuvant therapy) for different patients, which provide the possibility to improve survival. PMID- 18425578 TI - Ecotoxicity of engineered nanoparticles to aquatic invertebrates: a brief review and recommendations for future toxicity testing. AB - Based on a literature review and an overview of toxic effects of engineered nanoparticles in aquatic invertebrates, this paper proposes a number of recommendations for the developing field of nanoecotoxicology by highlighting the importance of invertebrates as sensitive and relevant test organisms. Results show that there is a pronounced lack of data in this field (less than 20 peer reviewed papers are published so far), and the most frequently tested engineered nanoparticles in invertebrate tests are C(60), carbon nanotubes, and titanium dioxide. In addition, the majority of the studies have used Daphnia magna as the test organism. To date, the limited number of studies has indicated acute toxicity in the low mg l(-1) range and higher of engineered nanoparticles to aquatic invertebrates, although some indications of chronic toxicity and behavioral changes have also been described at concentrations in the high microg l(-1) range. Nanoparticles have also been found to act as contaminant carriers of co-existing contaminants and this interaction has altered the toxicity of specific chemicals towards D. magna. We recommend that invertebrate testing is used to advance the level of knowledge in nanoecotoxicology through standardized short-term (lethality) tests with invertebrates as a basis for investigating behaviour and bioavailability of engineered nanoparticles in the aquatic environment. Based on this literature review, we further recommend that research is directed towards invertebrate tests employing long-term low exposure with chronic endpoints along with more research in bioaccumulation of engineered nanoparticles in aquatic invertebrates. PMID- 18425577 TI - Molecular profiles of progesterone receptor loss in human breast tumors. AB - BACKGROUND: Patient prognosis and response to endocrine therapy in breast cancer correlate with protein expression of both estrogen receptor (ER) and progesterone receptor (PR), with poorer outcome in patients with ER+/PR- compared to ER+/PR+ tumors. METHODS: To better understand the underlying biology of ER+/PR- tumors, we examined RNA expression (n > 1000 tumors) and DNA copy number profiles from five previously published studies of human breast cancers with clinically assigned hormone receptor status (ER+/PR+, ER+/PR-, and ER-/PR-). RESULTS: We identified an expression "signature" of genes with either elevated or diminished RNA levels specifically in ER+/PR+ compared to ER-/PR- and ER+/PR- tumors. We similarly identified a gene signature specific to ER-/PR- tumors. ER+/PR- tumors, on the other hand, were a mixture of three different subtypes: tumors manifesting the ER+/PR+ signature, tumors manifesting the ER-/PR- signature, and tumors not associating with ER+/PR+ or ER-/PR- tumors (which we considered "true" ER+/PR-). In analyses of both tamoxifen-treated and untreated patients, ER+/PR- breast cancers defined by RNA profiling were associated with poor patient outcome, worse than those with pure ER+/PR+ patterns; these differences were not observed when using clinical assays to assign ER and PR status. ER+/PR- tumors also showed twice as many DNA copy number gains or losses compared to ER+/PR+ and ER-PR- tumors. Targets of transcriptional up-regulation by specific oncogenic pathways, including PI3 K/Akt/mTOR, were enriched in both ER+/PR- and ER-/PR- compared to ER+/PR+ tumors. CONCLUSION: ER+/PR- tumors as defined by RNA profiling represent a distinct subset of breast cancer with aggressive features and poor outcome, despite being clinically ER+. Multigene assays derived from our gene signatures could conceivably provide an improved clinical assay for inferring PR status for prognostic and therapeutic purposes. PMID- 18425579 TI - Reversible secretion of glycosaminoglycans and proteoglycans by cyclically stretched valvular cells in 3D culture. AB - Mitral valve leaflets and chordae have been shown to contain different amounts and proportions of glycosaminoglycans (GAGs) and proteoglycans (PGs) corresponding to in vivo normal or diseased cyclic strain patterns. To understand the effect of cyclic strains on GAG/PG synthesis by valvular interstitial cells (VICs) isolated from valve leaflet and chordae separately, porcine VICs were seeded within collagen gels and alternately stretched or relaxed for 24 h periods for one week in a custom-designed tissue engineering bioreactor. We found cyclic stretch-induced upregulation of total GAGs and of individual GAG classes secreted into the culture medium. Leaflet cells showed a delayed response to stretching compared to chordal cells, but altered the proportions of various GAG classes they secreted during the culture duration. Decorin and biglycan PGs were slightly responsive to stretch. We demonstrated that mechanical stretch and relaxation conditions reversibly regulate GAG and PG production in a novel 3D model of valve tissues. This is the first study using cyclic strains to modulate GAG/PG synthesis by valve cells and our results may have implications for the remodeling of the mitral valve as well as other tissues. PMID- 18425580 TI - Analog electrical model of the coronary circulation in case of multiple revascularizations. AB - In this work, we propose an analog electrical model of the coronary circulation for patients with obstructive disease undergoing revascularization. In this clinical situation, the collateral circulation to the occluded artery is difficult to ascertain via preoperative measurements and well-developed collaterals might induce long-term restenosis of the revascularized artery due to flow competition mechanisms. The proposed model allows an original biomechanical analysis of per-operative hemodynamic data in order to assess quantitative evaluation of pressures and flows inside the native stenosed arteries, the collateral network and the bypass grafts. Average cardiac cycle values are analysed. In the case of 3-vessel disease and chronic occlusion of the right coronary artery, the quantitative results confirm the protective effects of the collateral flows in the pathological situation, but also show that the revascularization of the occluded right artery is fully justified since the collateral flows remain low, even when the left territory is revascularized. The model thus provides a computational tool to evaluate therapeutic strategies for each patient. PMID- 18425581 TI - In vivo validation of a realistic kinematic model for the trapezio-metacarpal joint using an optoelectronic system. AB - This article analyzes a realistic kinematic model of the trapezio-metacarpal (TM) joint in the human thumb that involves two non-orthogonal and non-intersecting rotation axes. The estimation of the model parameters, i.e. the position and orientation of the two axes with respect to an anatomical coordinate system, was carried out by processing the motion of nine retroreflective markers, externally attached to the hand surface, surveyed by a video motion capture system. In order to compute the model parameters, prototypical circumduction movements were processed within an evolutionary optimization approach. Quality and reproducibility in assessing the parameters were demonstrated across multiple testing sessions on 10 healthy subjects (both left and right thumbs), involving the complete removal of all markers and then retesting. Maximum errors of less than 5 mm in the axis position and less than 6 degrees in the orientation were found, respectively. The inter-subject mean distance between the two axes was 4.16 and 4.71 mm for right and left TM joints, respectively. The inter-subject mean relative orientation between the two axes was about 106 and 113 degrees for right and left TM joints, respectively. Generalization properties of the model were evaluated quantitatively on opposition movements in terms of distance between measured and predicted marker positions (maximum error less than 5 mm). The performance of the proposed model compared favorably with the one (maximum error in the range of 7-8 mm) obtained by applying a universal joint model (orthogonal and intersecting axes). The ability of in vivo estimating the parameters of the proposed kinematic model represents a significant improvement for the biomechanical analysis of the hand motion. PMID- 18425582 TI - Optimizing delivery of multivalent targeting constructs for detection of secondary tumors. AB - Targeting drugs or imaging molecules to specific cells by conjugating them to antibodies or ligands for cell surface receptors may allow earlier detection of pathology, better localization for intervention, and fewer side effects. Delivery of these molecules to the target is complicated by construct size, which cannot cross typical endothelial barriers such as the vascular wall, and lack of a priori knowledge of the location of secondary tumor sites to which the construct is targeted. Here we develop mathematical models for diffusive and convection enhanced delivery of a trivalent construct. Results show that delivery of the construct to the tissue does not yield acceptable contrast or specificity; therefore, unbound construct must be removed from the area of interest by allowing diffusion out of the area or a follow-up injection of fluid containing no construct (e.g., saline). The need for this additional step requires weeks to months for diffusive delivery to yield acceptable contrast, but convection enhanced delivery may be able to achieve acceptable contrast within several days. Thus, convection-enhanced delivery of multivalent constructs may provide a mechanism to locate secondary tumor sites without prior knowledge of their location which would greatly enhance the ability to detect and treat cancer. PMID- 18425583 TI - The GnRH test in the assessment of patients with pituitary and parapituitary lesions: results of a 5-year retrospective study. AB - OBJECTIVE: To examine the utility of the GnRH (gonadotrophin-releasing hormone) test in the management of patients with pituitary and parapituitary lesions. PATIENTS AND METHODS: A 5-year retrospective study of LH (luteinizing hormone) and FSH (follicle stimulating hormone) responses to GnRH test in patients with HP (hypothalamic-pituitary) disease in a regional endocrine centre. Serum LH and FSH concentrations were measured at baseline and at 20 and 60 min after an intravenous bolus of 100 mcg (micrograms) of GnRH. The GnRH responses were categorised by tumour size, tumour type, and gonadal status. RESULTS: Of the 104 patients studied, 46 were male and 58 were female. There were 50 normal, 38 subnormal and 16 exaggerated LH responses compared with 34 normal 67 subnormal and three exaggerated responses for FSH. Seventy-four patients (71.2%) were hypogonadal. Normal LH responses were achieved in half of the hypogonadal subjects and normal FSH responses in more than a third. Furthermore, the LH responses were exaggerated in nine hypogonadal patients compared with three for FSH. The GnRH test could not differentiate between pituitary or parapituitary lesions either by size or type of lesion. An exception was the male non functioning adenoma (NFA) sub-group (10 patients, all were hypopituitary, seven were hypogonadal), which demonstrated significant subnormal LH and FSH responses compared with other male and female tumour type sub-groups. CONCLUSIONS: The data from this study indicate that the GnRH test is unhelpful in the clinical assessment of the HP axis in patients with HP disease. PMID- 18425584 TI - Growth hormone deficiency in children. AB - The foundation for the diagnosis of growth hormone (GH) deficiency in childhood must be auxology, that is, the comparison of the child's growth pattern to that of established norms for gender and ethnicity. It is only in those growing considerably more slowly than average that testing for GHD makes sense. Assessment of laboratory tests, whether static, for example, the measurement of growth factors or their binding proteins, or dynamic, for example, secretagogue stimulated GH secretion is confirmatory. One must be cognizant of the assay used to determine GH, for there may be a 3-fold difference in the concentration of GH among commercially-available assays. Controversy still exists concerning the measurement of spontaneous GH release and whether sex-steroid priming is appropriate in prepubertal children. Imaging analysis may prove helpful in some children with congenital GHD or to detect a space-occupying lesion in the area of the hypothalamus and pituitary. The final diagnosis is based on multiple parameters and occasionally on a therapeutic trial of GH therapy to determine if there is a significant acceleration of growth velocity. PMID- 18425585 TI - Solution structure of wild-type human matrix metalloproteinase 12 (MMP-12) in complex with a tight-binding inhibitor. PMID- 18425586 TI - Christian ethics and the ethics of contemporary man. PMID- 18425587 TI - Catholic healthcare organizations and the articulation of their identity. PMID- 18425588 TI - Politics, bioethics, and science policy. PMID- 18425589 TI - Collaboration of ethics and patient safety programs: opportunities to promote quality care. PMID- 18425590 TI - A five year follow-up national study of ethics committees in medical organizations in Japan. PMID- 18425591 TI - Analysis of Arabidopsis arginase gene transcription patterns indicates specific biological functions for recently diverged paralogs. AB - The detailed expression patterns of transcripts of two Arabidopsis arginase genes, ARGAH1 and ARGAH2, have not been previously described, and phylogenetic analysis suggests that they diverged independently of duplication events in other lineages. Therefore, we used beta-glucuronidase reporter fusions and quantitative reverse-transcriptase PCR to analyze tissue-specific expression of ARGAH1 and ARGAH2 during Arabidopsis development, and in response to the availability of nutrients and exposure to methyl jasmonate (MeJA). We demonstrated tissue specific transcript expression and enzyme activity in pollen for ARGAH1, but not ARGAH2. Conversely, we demonstrated MeJA-inducibility of ARGAH2, but not ARGAH1. In addition, we used microarrays to identify genes for which transcript abundance following MeJA treatment differed in wild type and ARGAH2 mutants. These ARGAH2 and MeJA responsive genes included a putative pathogenesis-related protein pathogenesis response-1 (At2g14610), and a gene of unknown function (At5g03090). Interestingly, these genes had opposite responses to the loss of ARGAH2, suggesting multiple downstream effects of arginase activity, following MeJA treatment. These results, and the variety and complexity of expression patterns of ARGAH1 and ARGAH2 transcript expression and their related reporter gene fusions that we observed point to multiple functions of arginase genes in Arabidopsis, some of which have resulted through a sub-functionalization not shared by all angiosperms. PMID- 18425592 TI - Collaborative pooled analysis of data on C-reactive protein gene variants and coronary disease: judging causality by Mendelian randomisation. AB - Many prospective studies have reported associations between circulating C reactive protein (CRP) levels and risk of coronary heart disease (CHD), but causality remains uncertain. Studies of CHD are being conducted that involve measurement of common polymorphisms of the CRP gene known to be associated with circulating concentrations, thereby utilising these variants as proxies for circulating CRP levels. By analysing data from several studies examining the association between relevant CRP polymorphisms and CHD risk, the present collaboration will undertake a Mendelian randomisation analysis to help assess the likelihood of any causal relevance of CRP levels to CHD risk. A central database is being established containing individual data on CRP polymorphisms, circulating CRP levels, and major coronary outcomes as well as age, sex and other relevant characteristics. Associations between CRP polymorphisms or haplotypes and CHD will be evaluated under different circumstances. This collaboration comprises, at present, about 37,000 CHD outcomes and about 120,000 controls, which should yield suitably precise findings to help judge causality. This work should advance understanding of the relevance of low-grade inflammation to CHD and indicate whether or not CRP itself is involved in long-term pathogenesis. PMID- 18425594 TI - Evaluation of early embrionic development after natural mating using ultrasonography in bos indicus cows in the humid tropics of Costa Rica. AB - Fifty-eight nursing Zebu cows averaging 5922 days postpartum were used to evaluate the early embryonic development by ultrasound under a natural mating system (bulls A to F). Three consecutive mating periods of 21 days were established. Sixty two percent of the animals were diagnosed as pregnant (36/58). During the first period (AB) 76% (27/36) of the animals become pregnant, 17% (6/36) during the second (CD) and 9% (3/36) in the last mating period (EF). Sixty two percent of the animals were true positives (animals found pregnant diagnosed by ultrasound (US), and conforming with the embryonic scale proposed by Rosiles et al. (2006); 12% (n = 7) of false negatives (pregnant cows by US but not conforming with the scale); 2% (n = 1) of false positives (pregnant cows, evaluated with US and conforming with the embryonic scale but not pregnant at the end of the study); and a 26% (n = 15) of true negatives (not pregnancy evaluated with US). The sensitivity of the test was 97%, with a specificity of 68% with a positive predictive value of 83% and a negative predictive value of 6%. Relation between the measuring scale of embryo development and the results obtained by ultrasonography was k = 0.69. No significant correlation were observed between body condition score (BCS) evaluation and body fat (BF) measurement at any of the three evaluations performed after mating, however, measurements of BCS and BF among time showed a positive correlation (0.48 P < 0.05, 0.29, P < 0.07, respectively) from the beginning of the evaluation period to the end of the trial. Ultrasonography to monitor embryonic development to determine the early establishment of pregnancy in natural mating reproductive programs can be a valuable technique to monitor reproductive efficiency. PMID- 18425593 TI - Force transients and minimum cross-bridge models in muscular contraction. AB - Two- and three-state cross-bridge models are considered and examined with respect to their ability to predict three distinct phases of the force transients that occur in response to step change in muscle fiber length. Particular attention is paid to satisfying the Le Chatelier-Brown Principle. This analysis shows that the two-state model can account for phases 1 and 2 of a force transient, but is barely adequate to account for phase 3 (delayed force) unless a stretch results in a sudden increase in the number of cross-bridges in the detached state. The three-state model (A-->B-->C-->A) makes it possible to account for all three phases if we assume that the A-->B transition is fast (corresponding to phase 2), the B-->A transition is of intermediate speed (corresponding to phase 3), and the C-->A transition is slow; in such a scenario, states A and C can support or generate force (high force states) but state B cannot (detached, or low-force state). This model involves at least one ratchet mechanism. In this model, force can be generated by either of two transitions: B-->A or B-->C. To determine which of these is the major force-generating step that consumes ATP and transduces energy, we examine the effects of ATP, ADP, and phosphate (Pi) on force transients. In doing so, we demonstrate that the fast transition (phase 2) is associated with the nucleotide-binding step, and that the intermediate-speed transition (phase 3) is associated with the Pi-release step. To account for all the effects of ligands, it is necessary to expand the three-state model into a six-state model that includes three ligand-bound states. The slowest phase of a force transient (phase 4) cannot be explained by any of the models described unless an additional mechanism is introduced. Here we suggest a role of series compliance to account for this phase, and propose a model that correlates the slowest step of the cross-bridge cycle (transition C-->A) to: phase 4 of step analysis, the rate constant k(tr) of the quick-release and restretch experiment, and the rate constant k(act) for force development time course following Ca(2+) activation. PMID- 18425595 TI - Interleukin-1beta infusion in bovine mammary glands prior to challenge with Streptococcus uberis reduces bacterial growth but causes sterile mastitis. AB - Dairy cows are especially vulnerable to intramammary infection by the bacterial pathogen Streptococcus uberis in the dry period. Use of immunotherapeutic agents at drying off could increase cellular defences in the gland and prevent establishment of new S. uberis infections. This study investigated the potential of infusing recombinant bovine interleukin-1 beta (rbIL-1beta) in the mammary glands as a prophylactic agent against subsequent intramammary challenge with S. uberis in the early dry period. Immediately after the last milking at commencement of the dry period, one cow from each of 10 monozygous twinsets was infused with 10 microg of rbIL-1beta in two quarters and the other twin was infused with the carrier agent, sterile phosphate buffered saline. Twenty-four hours later, the quarters were infused with 10(3) colony-forming units (CFU) of S. uberis. Bacteriology, somatic cell count (SCC), concentrations of specific cytokines and antibody responses were monitored in mammary gland secretions and sera for the next 21 days. Infusion of rbIL-1beta into mammary glands at commencement of the dry period was associated with less new S. uberis intramammary infections, as determined by the number of quarters with bacterial growth. However, high SCC in quarters following infusion of rbIL-1beta masked the full beneficial effect of this procedure. PMID- 18425596 TI - DNA vaccines and their applications in veterinary practice: current perspectives. AB - Inoculation of plasmid DNA, encoding an immunogenic protein gene of an infectious agent, stands out as a novel approach for developing new generation vaccines for prevention of infectious diseases of animals. The potential of DNA vaccines to act in presence of maternal antibodies, its stability and cost effectiveness and the non-requirement of cold chain have heightened the prospects. Even though great strides have been made in nucleic acid vaccination, still there are many areas that need further research for its wholesome practical implementation. Major areas of concern are vaccine delivery, designing of suitable vectors and cytotoxic T cell responses. Also, the induction of immune responses by DNA vaccines is inconclusive due to the lack of knowledge regarding the concentration of the protein expressed in vivo. Alternative delivery systems having higher transfection efficiency and the use of cytokines, as immunomodulators, needs to be further explored. Recently, efforts are being made to modulate and prolong the active life of dendritic cells, in order to make antigen presentation a more efficacious one. For combating diseases like acquired immunodeficiency syndrome (AIDS), influenza, malaria and tuberculosis in humans; and foot and mouth disease, Aujesky's disease, swine fever, rabies, canine distemper and brucellosis in animals, DNA vaccine clinical trials are underway. This review highlights the salient features of DNA vaccines, and measures to enhance their efficacy so as to devise an effective and novel vaccination strategy against animal diseases. PMID- 18425597 TI - Two new ascomycetous anamorphic yeast species related to Candida friedrichii- Candida jaroonii sp. nov., and Candida songkhlaensis sp. nov.--isolated in Thailand. AB - In a study of yeast diversity in Thailand, eight strains of hitherto undescribed anamorphic yeasts were isolated: four from insect frass, two from Marasmius sp. fruiting bodies, one from a flower, and one from jackfruit exudates. Phylogenetic analysis of the D1/D2 domain of 26S ribosomal DNA nucleotide sequences indicated that the eight strains represented two new species related to Candida friedrichii. Genetic separation of the two new species was further supported by DNA-DNA hybridization analysis, which resulted in between-species similarity values of less than 48%, and by electrophoretic karyotyping. The two new species are C. jaroonii sp. nov. (type strain, ST-300(T) = NBRC 103209(T) = BCC 11783(T) = CBS 10790(T)) and C. songkhlaensis sp. nov. (type strain, ST-328(T) = NBRC 103214(T) = BCC 11804(T) = CBS 10791(T)). PMID- 18425598 TI - Isotopic labelling of photosystem II in Thermosynechococcus elongatus. AB - This report describes a protocol to incorporate isotopically labelled aromatic amino acids into the proteins of the thermophilic cyanobacterium Thermosynechoccus elongatus. By using the EPR signal of the two redox active tyrosines of Photosystem II, Tyr(D)(*) and Tyr(Z)(*), as spectroscopic probes it is shown that labelled tyrosines can be incorporated with a high yield in this cyanobacterium. The production of a fully (13)C- or (2)H-labelled enzyme is also described. PMID- 18425599 TI - Herbicide effect on the hydrogen-bonding interaction of the primary quinone electron acceptor QA in photosystem II as studied by Fourier transform infrared spectroscopy. AB - The redox potential of Q(A) in photosystem II (PSII) is known to be lower by approximately 100 mV in the presence of phenolic herbicides compared with the presence of DCMU-type herbicides. In this study, the structural basis underlying the herbicide effects on the Q(A) redox potential was studied using Fourier transform infrared (FTIR) spectroscopy. Light-induced Q(A)(-)/Q(A) FTIR difference spectra of Mn-depleted PSII membranes in the presence of DCMU, atrazine, terbutryn, and bromacil showed a strong CO stretching peak of Q(A)(-) at 1,479 cm(-1), while binding of phenolic herbicides, bromoxynil and ioxynil, induced a small but clear downshift by approximately 1 cm(-1). The CO peak positions and the small frequency difference were reproduced in the S(2)Q(A)( )/S(1)Q(A) spectra of oxygen-evolving PSII membranes with DCMU and bromoxynil. The relationship of the CO frequency with herbicide species correlated well with that of the peak temperatures of thermoluminescence due to S(2)Q(A)(-) recombination. Density functional theory calculations of model hydrogen-bonded complexes of plastoquinone radical anion showed that the small shift of the CO frequency is consistent with a change in the hydrogen-bond structure most likely as a change in its strength. The Q(A)(-)/Q(A) spectra in the presence of bromoxynil, and ioxynil, which bear a nitrile group in the phenolic ring, also showed CN stretching bands around 2,210 cm(-1). Comparison with the CN frequencies of bromoxynil in solutions suggested that the phenolic herbicides take a phenotate anion form in the Q(B) pocket. It was proposed that interaction of the phenolic C-O(-) with D1-His215 changes the strength of the hydrogen bond between the CO of Q(A) with D2-His214 via the iron-histidine bridge, causing the decrease in the Q(A) redox potential. PMID- 18425600 TI - Molecular cloning and characterization of crustin from mud crab Scylla paramamosain. AB - Antimicrobial peptides (AMPs) are important components of the host innate immune response against microbial invasion. In the present study, we report the identification and characterization of a crustin (CrusSp) from the hemocyte of mud crab, Scylla paramamosain using an expressed sequence tag (EST) and rapid amplification cDNA end (RACE) approaches. Analysis of the nucleotide sequence revealed seven different variances of the CrusSp cDNA in mud crab. The open reading frame encodes a protein of 111 amino acids with 21 residues signal sequence. The predicted molecular mass of the mature protein (90 amino acids) is 10.27 kDa with an estimated pI of 8.54. Analysis of the protein domain features indicated typical conserved cysteine residues containing a single whey acidic protein (WAP) domain at the C-terminus. A neighbour-joining tree showed that S. paramamosain crustin is closely related to other crustin homologues, and displays the highest similarity to crustin antimicrobial peptide in shore crab Carcinus maenas. Four exons and three introns were identified within the 999 bp genomic DNA sequence of CrusSp. Tissue distribution analysis showed that CrusSp was highly expressed in hemocytes, gills, intestines and muscle but it was not expressed in hepatopancreas and eyestalks. To gain insight into the in vitro antimicrobial activities of CrusSp, the mature peptide coding region was cloned into E. coli for heterologous expression. The recombinant CrusSp could inhibit the growth of gram-positive bacteria but had no inhibition activity against gram negative bacteria. These results indicated the involvement of CrusSp in the innate immunity of S. paramamosain. PMID- 18425602 TI - New insights into the acute care of patients with aneurysmal subarachnoid hemorrhage. PMID- 18425601 TI - Engineering with uncertainty: monitoring air bag performance. AB - Modern engineering is complicated by an enormous number of uncertainties. Engineers know a great deal about the material world and how it works. But due to the inherent limits of testing and the complexities of the world outside the lab, engineers will never be able to fully predict how their creations will behave. One way the uncertainties of engineering can be dealt with is by actively monitoring technologies once they have left the development and production stage. This article uses an episode in the history of automobile air bags as an example of engineers who had the foresight and initiative to carefully track the technology on the road to discover problems as early as possible. Not only can monitoring help engineers identify problems that surface in the field, it can also assist them in their efforts to mobilize resources to resolve problem. PMID- 18425603 TI - Fulminant hepatic failure: when the hepatologist meets the neurointensivist. PMID- 18425606 TI - High-resolution thermogravimetric analysis for rapid characterization of biomass composition and selection of shrub willow varieties. AB - The cultivation of shrub willow (Salix spp.) bioenergy crops is being commercialized in North America, as it has been in Europe for many years. Considering the high genetic diversity and ease of hybridization, there is great potential for genetic improvement of shrub willow through traditional breeding. The State University of New York-College of Environmental Science and Forestry has an extensive breeding program for the genetic improvement of shrub willow for biomass production and for other environmental applications. Since 1998, breeding efforts have produced more than 200 families resulting in more than 5,000 progeny. The goal for this project was to utilize a rapid, low-cost method for the compositional analysis of willow biomass to aid in the selection of willow clones for improved conversion efficiency. A select group of willow clones was analyzed using high-resolution thermogravimetric analysis (HR-TGA), and significant differences in biomass composition were observed. Differences among and within families produced through controlled pollinations were observed, as well as differences by age at time of sampling. These results suggest that HR-TGA has a great promise as a tool for rapid biomass characterization. PMID- 18425604 TI - Light-dependent compartmentalization of transducin in rod photoreceptors. AB - Three major visual signaling proteins, transducin, arrestin, and recoverin undergo bidirectional translocations between the outer segment and inner compartments of rod photoreceptors in a light-dependent manner. The light dependent translocation of proteins is believed to contribute to adaptation and neuroprotection of photoreceptor cells. The potential physiological significance and mechanisms of light-controlled protein translocations are at the center of current discussion. In this paper, I outline the latest advances in understanding the mechanisms of bidirectional translocation of transducin and determinants of its steady-state distribution in dark- and light-adapted photoreceptor cells. PMID- 18425607 TI - Assessment of bermudagrass and bunch grasses as feedstock for conversion to ethanol. AB - Research is needed to allow more efficient processing of lignocellulose from abundant plant biomass resources for production to fuel ethanol at lower costs. Potential dedicated feedstock species vary in degrees of recalcitrance to ethanol processing. The standard dilute acid hydrolysis pretreatment followed by simultaneous sacharification and fermentation (SSF) was performed on leaf and stem material from three grasses: giant reed (Arundo donax L.), napiergrass (Pennisetum purpureum Schumach.), and bermudagrass (Cynodon spp). In a separate study, napiergrass, and bermudagrass whole samples were pretreated with esterase and cellulose before fermentation. Conversion via SSF was greatest with two bermudagrass cultivars (140 and 122 mg g(-1) of biomass) followed by leaves of two napiergrass genotypes (107 and 97 mg g(-1)) and two giant reed clones (109 and 85 mg g(-1)). Variability existed among bermudagrass cultivars for conversion to ethanol after esterase and cellulase treatments, with Tifton 85 (289 mg g) and Coastcross II (284 mg g(-1)) being superior to Coastal (247 mg g(-1)) and Tifton 44 (245 mg g(-1)). Results suggest that ethanol yields vary significantly for feedstocks by species and within species and that genetic breeding for improved feedstocks should be possible. PMID- 18425605 TI - Commentary: "re-programming or selecting adult stem cells?". AB - The recent observations that embryonic stemness-associated genes could assist in the "de-differentiation" of adult skin fibroblast cells to "embryonic-like stem cells", using the "somatic cell nuclear transfer" techniques, have been interpreted as indicating a "re-programming" of genes. These reports have demonstrated a "proof of principle" approach to by-pass many, but not all, of the ethical, scientific and medical limitations of the "therapeutic cloning" of embryonic stem cells from embryos. However, while the interpretation that real "re-programming" of all those somatic fibroblastic differentiation genes might be correct, there does exists an alternative hypothesis of these exciting results. Based on the fact that multipotent adult stem cells exist in most, if not all, adult organs, the possibility exists that all these recent "re-programming" results, using the somatic nuclear transfer techniques, actually were the results of transferred rare nuclear material from the adult stem cells residing in the skin of the mouse, monkey and human samples. An examination of the rationale for this challenging hypothesis has been drawn from the hypothesis of the "stem cell theory of cancer", as well as from the field of human adult stem cells research. PMID- 18425608 TI - Rapid isolation of the Trichoderma strain with higher degrading ability of a filter paper and superior proliferation characteristics using avicel plates and the double-layer selection medium. AB - The cost of cellulase is still a problem for bioethanol production. As the cellulase of Trichoderma reesei is applicable for producing ethanol from cellulosic materials, the cellulase productivity of this fungus should be increased. Therefore, we attempted to develop a system to isolate the strain with higher degrading ability of a filter paper and superior proliferation characteristics among the conidia treated with the mitotic arrester, colchicine. When green mature conidia of T. reesei RUT C-30 were swollen, autopolyploidized, and incubated in the double-layer selection medium containing Avicel, colonies appeared on the surface earlier than the original strain. When such colonies and the original colony were incubated on the Avicel plates, strain B5, one of the colonies derived from the colchicine-treated conidia, showed superior proliferation characteristics. Moreover, when strain B5 and the original strain were compared in the filter paper degrading ability and the cellulose hydrolyzing activity, strain B5 was also superior to the original strain. It was suspected that superior proliferation characteristics of strain B5 reflects higher filter paper degrading ability. Thus, we concluded that the Trichoderma strain with higher degrading ability of a filter paper and superior proliferation characteristics can be isolated using Avicel plates and the double-layer selection medium. PMID- 18425609 TI - A comparison of simple rheological parameters and simulation data for Zymomonas mobilis fermentation broths with high substrate loading in a 3-L bioreactor. AB - Traditionally, as much as 80% or more of an ethanol fermentation broth is water that must be removed. This mixture is not only costly to separate but also produces a large aqueous stream that must then be disposed of or recycled. Integrative approaches to water reduction include increasing the biomass concentration during fermentation. In this paper, experimental results are presented for the rheological behavior of high-solids enzymatic cellulose hydrolysis and ethanol fermentation for biomass conversion using Solka Floc as the model feedstock. The experimental determination of the viscosity, shear stress, and shear rate relationships of the 10 to 20% slurry concentrations with constant enzyme concentrations are performed with a variable speed rotational viscometer (2.0 to 200 rpm) at 40 degrees C. The viscosities of enzymatic suspension observed were in range of 0.0418 to 0.0144, 0.233 to 0.0348, and 0.292 to 0.0447 Pa s for shear rates up to 100 reciprocal seconds at 10, 15, and 20% initial solids (w/v), respectively. Computational fluid dynamics analysis of bioreactor mixing demonstrates the change in bioreactor mixing with increasing biomass concentration. The portion-loading method is shown to be effective for processing high-solids slurries. PMID- 18425610 TI - Effects of oxygen limitation on xylose fermentation, intracellular metabolites, and key enzymes of Neurospora crassa AS3.1602. AB - The effects of oxygen limitation on xylose fermentation of Neurospora crassa AS3.1602 were studied using batch cultures. The maximum yield of ethanol was 0.34 g/g at oxygen transfer rate (OTR) of 8.4 mmol/L.h. The maximum yield of xylitol was 0.33 g/g at OTR of 5.1 mmol/L.h. Oxygen limitation greatly affected mycelia growth and xylitol and ethanol productions. The specific growth rate (micro) decreased 82% from 0.045 to 0.008 h(-1) when OTR changed from 12.6 to 8.4 mmol/L.h. Intracellular metabolites of the pentose phosphate pathway, glycolysis, and tricarboxylic acid cycle were determined at various OTRs. Concentrations of most intracellular metabolites decreased with the increase in oxygen limitation. Intracellular enzyme activities of xylose reductase, xylitol dehydrogenase, and xylulokinase, the first three enzymes in xylose metabolic pathway, decreased with the increase in oxygen limitation, resulting in the decreased xylose uptake rate. Under all tested conditions, transaldolase and transketolase activities always maintained at low levels, indicating a great control on xylose metabolism. The enzyme of glucose-6-phosphate dehydrogenase played a major role in NADPH regeneration, and its activity decreased remarkably with the increase in oxygen limitation. PMID- 18425611 TI - Fermentation of acid-pretreated corn stover to ethanol without detoxification using Pichia stipitis. AB - In this work, the effect of adaptation on P. stipitis fermentation using acid pretreated corn stover hydrolyzates without detoxification was examined. Two different types of adaptation were employed, liquid hydrolyzate and solid state agar adaptation. Fermentation of 12.5% total solids undetoxified acid-pretreated corn stover was performed in shake flasks at different rotation speeds. At low rotation speed (100 rpm), both liquid hydrolyzate and solid agar adaptation highly improved the sugar consumption rate as well as ethanol production rate compared to the wild-type strains. The fermentation rate was higher for solid agar-adapted strains compared to liquid hydrolyzate-adapted strains. At a higher rotation speed (150 rpm), there was a faster sugar consumption and ethanol production for both the liquid-adapted and the wild-type strains. However, improvements in the fermentation rate between the liquid-adapted and wild strains were less pronounced at the high rotation speed. PMID- 18425612 TI - Bioethanol production from uncooked raw starch by immobilized surface-engineered yeast cells. AB - Surface-engineered yeast Saccharomyces cerevisiae codisplaying Rhizopus oryzae glucoamylase and Streptococcus bovis alpha-amylase on the cell surface was used for direct production of ethanol from uncooked raw starch. By using 50 g/L cells during batch fermentation, ethanol concentration could reach 53 g/L in 7 days. During repeated batch fermentation, the production of ethanol could be maintained for seven consecutive cycles. For cells immobilized in loofa sponge, the concentration of ethanol could reach 42 g/L in 3 days in a circulating packed-bed bioreactor. However, the production of ethanol stopped thereafter because of limited contact between cells and starch. The bioreactor could be operated for repeated batch production of ethanol, but ethanol concentration dropped to 55% of its initial value after five cycles because of a decrease in cell mass and cell viability in the bioreactor. Adding cells to the bioreactor could partially restore ethanol production to 75% of its initial value. PMID- 18425614 TI - Bioreactors for H2 production by purple nonsulfur bacteria. AB - Two types of laboratory-scale bioreactors were designed for H(2) production by purple nonsulfur bacteria. The bioreactors employed a unique type of hydrogenase activity found in some photosynthetic bacteria that functions in darkness to shift CO (and H2O) into H(2) (and CO2). The mass transport of gaseous CO into an aqueous bacterial suspension was the rate-limiting step and the main challenge for bioreactor design. Hollow-fiber and bubble-train bioreactors employing immobilized and free-living bacteria have proven effective for enhancing the mass transfer of CO. The hollow-fiber bioreactor was designed so that both a growth medium and CO (10% in N(2)) passed from the inside of the fibers to the outside within the bioreactor. Bacteria were immobilized on the outer surface of the hollow fibers. Hydrogen production from CO at an average rate of 125 ml g cdw(-1) h(-1) (maximum rate of 700 ml g cdw(-1) h(-1)) was observed for more than 8 months. The bubble-train bioreactor was built using polyvinyl chloride (PVC) tubing, wound helically on a vertical cylindrical supporting structure. Small bubbles containing CO were injected continuously through a needle/septum connection from the gas reservoir (20% CO). Up to 140 ml g cdw(-1) h(-1) of H(2) production activity was observed using this bioreactor for more than 10 days. PMID- 18425613 TI - Effects of gene orientation and use of multiple promoters on the expression of XYL1 and XYL2 in Saccharomyces cerevisiae. AB - Orientation of adjacent genes has been reported to affect their expression in eukaryotic systems, and metabolic engineering also often makes repeated use of a few promoters to obtain high expression. To improve transcriptional control in heterologous expression, we examined how these factors affect gene expression and enzymatic activity in Saccharomyces cerevisiae. We assembled D-xylose reductase (XYL1) and D-xylitol dehydrogenase (XYL2) in four ways. Each pair of genes was placed in two different tandem (1-->2--> or <--1<--2), convergent (1--><--2), and divergent (<--1 2-->) orientations in autonomous plasmids. The TEF1 promoter was used to drive XYL1 and the TDH3 promoter to drive XYL2 in each of the constructs. The effects of gene orientation on growth, transcription, and enzyme activity were analyzed. The transcription level as measured by quantitative PCR (q-PCR) correlated with enzyme activities, but our data did not show a significant effect of gene orientation. To test the possible dilution of promoter strength due to multiple use of the same promoter, we examined the level of expression of XYL1 driven by either the TEF1 or TDH3 promoter when carried on a single copy plasmid. We then co-expressed XYL2 from either a single or multicopy plasmid, which was also driven by the same promoter. XYL2 transcript and enzyme expression increased with plasmid copy number, while the expression of XYL1 was constant regardless of the number of other TEF1 or TDH3 promoters present in the cell. According to our data, there is no significant effect of gene orientation or multiple promoter use on gene transcription and translation when genes are expressed from plasmids; however, other factors could affect expression of adjacent genes in chromosomes. PMID- 18425615 TI - Solid-state fermentation of xylanase from Penicillium canescens 10-10c in a multi layer-packed bed reactor. AB - Xylanase is produced by Penicillium canescens 10-10c from soya oil cake in static conditions using solid-state fermentation. The impact of several parameters such as the nature and the size of inoculum, bed-loading, and aeration is evaluated during the fermentation process. Mycelial inoculum gives more production than conidial inoculum. Increasing the quantity of inoculum enhances slightly xylanase production. Forced aeration induces more sporulation of strain and reduces xylanase production. However, forced moistened air improves the production compared to production obtained with forced dry air. In addition, increasing bed loading reduces the specific xylanase production likely due to the incapacity of the Penicillium strain to grow deeply in the fermented soya oil cake mass. Thus, the best cultivation conditions involve mycelial inoculum form, a bed loading of 1-cm height and passive aeration. The maximum xylanase activity is obtained after 7 days of fermentation and attains 10,200 U/g of soya oil cake. These levels are higher than those presented in the literature and, therefore, show all the potentialities of this stock and this technique for the production of xylanase. PMID- 18425616 TI - Ethanol production from wet-exploded wheat straw hydrolysate by thermophilic anaerobic bacterium Thermoanaerobacter BG1L1 in a continuous immobilized reactor. AB - Thermophilic ethanol fermentation of wet-exploded wheat straw hydrolysate was investigated in a continuous immobilized reactor system. The experiments were carried out in a lab-scale fluidized bed reactor (FBR) at 70 degrees C. Undetoxified wheat straw hydrolysate was used (3-12% dry matter), corresponding to sugar mixtures of glucose and xylose ranging from 12 to 41 g/l. The organism, thermophilic anaerobic bacterium Thermoanaerobacter BG1L1, exhibited significant resistance to high levels of acetic acid (up to 10 g/l) and other metabolic inhibitors present in the hydrolysate. Although the hydrolysate was not detoxified, ethanol yield in a range of 0.39-0.42 g/g was obtained. Overall, sugar efficiency to ethanol was 68-76%. The reactor was operated continuously for approximately 143 days, and no contamination was seen without the use of any agent for preventing bacterial infections. The tested microorganism has considerable potential to be a novel candidate for lignocellulose bioconversion into ethanol. The work reported here also demonstrates that the use of FBR configuration might be a viable approach for thermophilic anaerobic ethanol fermentation. PMID- 18425617 TI - Succinic acid production from cheese whey using Actinobacillus succinogenes 130 Z. AB - Actinobacillus succinogenes 130 Z was used to produce succinic acid from cheese whey in this study. At the presence of external CO(2) supply, the effects of initial cheese whey concentration, pH, and inoculum size on the succinic acid production were studied. The by-product formation during the fermentation process was also analyzed. The highest succinic acid yield of 0.57 was obtained at initial cheese whey concentration of 50 g/L, while the highest succinic acid productivity of 0.58 g h(-1) L(-1) was obtained at initial cheese whey concentration of 100 g/L. Increase in pH and inoculum size caused higher succinic acid yield and productivity. At the preferred fermentation condition of pH 6.8, inoculum size of 5% and initial cheese whey concentration of 50 g/L, succinic acid yield of 0.57, and productivity of 0.44 g h(-1) L(-1) were obtained. Acetic acid and formic acid were the main by-products throughout the fermentation run of 48 h. It is feasible to produce succinic acid using lactose from cheese whey as carbon resource by A. succinogenes 130 Z. PMID- 18425618 TI - Altered KCNQ3 potassium channel function caused by the W309R pore-helix mutation found in human epilepsy. AB - The second tryptophan (W) residue of the conserved WW motif in the pore helix of many K+ channel subunit is thought to interact with the tyrosine (Y) residues of the selectivity filter. A missense mutation causing the replacement of the corresponding residues with an arginine (W309R) occurs in KCNQ3 subunits forming part of M-channels. In this study, we examined the functional consequences of the W309R mutation in heterogously expressed KCNQ channels. Homomeric KCNQ3W309R channels lacked KCNQ currents. Heteromeric KCNQ2/KCNQ3W309R channels displayed a dominant-negative suppression of current and a significant modification in gating properties when compared with heteromeric KCNQ3/KCNQ2 channels mimicking the M channels. A three-dimensional homology model in the W309R mutant indicated that the R side chain of pore helices is too far from the Y side chain of the selectivity filter to interact via hydrogen bonds with each other and stabilize the pore structure. Collectively, the present results suggest that the second W residues of pore helices and their chemical interaction with the Y residues of the selectivity filter are essential for normal K+ channel function. This pore helix mutation, if occurs in the brain M channels, could thus lead to a channel dysfunction sufficient to trigger epileptic hyperexcitability. PMID- 18425619 TI - [Isometric grip strength and social gerontological research: results and analytic potentials of SHARE and SOEP]. AB - This paper shows that the measurement of hand grip strength provides a non invasive and reliable objective health indicator for social science research and is easy to collect in general population surveys. Grip strength is not only a useful complement of self-reported indicators of health, but it also exhibits a considerable predictive power with regard to a number of further relevant variables for social gerontological research, such as mortality risks. New data from the 2004 Survey of Health, Ageing and Retirement in Europe (SHARE) and the 2006 wave of the German Socio-Economic Panel Study (SOEP) allow insightful methodological and very first substantive cross-sectional analyses of grip strength in Germany. The focus of the present study is on the analysis of individuals aged 50 or older. The experience of both surveys when measuring grip strength is consistently positive, particularly with regard to the respondents' feedback. Major determinants of isometric grip strength are - beyond the individual's gender - age, body size and weight. A multivariate analysis also provides evidence for a clear positive association between various health indicators and grip strength. PMID- 18425620 TI - Cerebral involvement in axonal Charcot-Marie-Tooth neuropathy caused by mitofusin2 mutations. AB - Mutations in the mitofusin 2 (MFN2) gene are a major cause of primary axonal Charcot- Marie-Tooth (CMT) neuropathy. This study aims at further characterization of cerebral white matter alterations observed in patients with MFN2 mutations. Molecular genetic, magnetic resonance imaging (MRI), magnetic resonance spectroscopy (MRS), and diffusion tensor imaging (DTI) investigations were performed in four unrelated patients aged 7 to 38 years with early onset axonal CMT neuropathy. Three distinct and so far undescribed MFN2 mutations were detected. Two patients had secondary macrocephaly and mild diffuse predominantly periventricular white matter alterations on MRI. In addition, one boy had symmetrical T2-hyperintensities in both thalami. Two patients had optic atrophy, one of them with normal MRI. In three patients proton MRS revealed elevated concentrations of total N-acetyl compounds (neuronal marker), total creatine (found in all cells) and myo-inositol (astrocytic marker) in cerebral white and gray matter though with regional variation. These alterations were most pronounced in the two patients with abnormal MRI. DTI of these patients revealed mild reductions of fractional anisotropy and mild increase of mean diffusivity in white matter. The present findings indicate an enhanced cellular density in cerebral white matter of MFN2 neuropathy which is primarily due to a reactive gliosis without axonal damage and possibly accompanied by mild demyelination. PMID- 18425621 TI - Thalidomide causes sinus bradycardia in ALS. AB - OBJECTIVE: Neuroinflammation contributes to motor neuron degeneration in ALS. Thalidomide (THL) shows potent anti-inflammatory properties and increased the lifespan in ALS transgenic mice. Thalidomide was therefore suggested as atherapeutic intervention for the treatment of ALS.We conducted a pilot, randomized clinical trial of THL in patients with ALS to assess safety, feasibility, and preliminary estimates of treatment efficacy. METHODS: Patients were randomized to THL in combination with riluzole (n = 18) or riluzole alone (n = 19). THL was initiated at 100 mg per day for 6 weeks. Thereafter, the dose was increased every week by 50 mg until reaching the dose of 400 mg per day and planned to continue for another 12 weeks. RESULTS: Within 12 weeks of THL treatment, nine THL patients (50%) developed bradycardia defined as a heart rate below 60 beats per minute (bpm) and ranged from 46 to 59 bpm. Mean heart rate dropped by 17 bpm with THL treatment. Severe symptomatic bradycardia of 30 bpm occurred in one patient. A further patient died from sudden unexpected death. The study was terminated prematurely for safety concerns. The secondary outcome variables showed similar results for both groups. CONCLUSION: Bradycardia was the most common adverse event of THL treatment in ALS. THL-related bradycardia does not appear to be ALS-specific. It is conceivable, however, that the unexpected frequency and severity of THL-induced bradycardia may be related to subclinical involvement of the autonomic nervous system in ALS. The cardiac toxicity discourages further clinical trials and compassionate use of THL in ALS. ClinicalTrials.gov Identifier: NCT00231140. PMID- 18425622 TI - Comparison of two different measurement techniques of hippocampal resection length in temporal lobe epilepsy: results of a prospective study. AB - BACKGROUND: Magnetic resonance imaging (MRI) volumetry has evolved to a highly sensitive method for presurgical detection of hippocampal sclerosis in temporal lobe epilepsy (TLE). Seizure resolution and neuropsychological sequelae are believed to correlate with extent of resection. Therefore an easy volumetric method to determine extent of resection is desirable. The purpose of this work is to evaluate and compare two different measurement techniques for hippocampal resection length. METHODS: Sixty-one patients with a mean seizure history of 25.1 years and medically intractable TLE were included. They underwent MRI with sagittal acquired 3D T1-weighted spoiled gradient recalled echo sequence in 1 mm(3) isotropic voxel. Hippocampal resection length was calculated with two different methods. In the slice counting method (SCM) the number of consecutive 1 mm-thick slices containing resected hippocampus formation was counted. In the vector method (VM) the sum of the oblique and thus longer distances between the centre points of segmented hippocampal areas on each MRI slice were calculated. RESULTS: Since the hippocampus is a curved body, the resection lengths measured with VM were always larger than measured with SCM. The comparison of resection length expressed in "percent of total length" showed good agreement between the two methods, because unlike the absolute values of resection length, the percentage values are unaffected by the three-dimensional shape of the hippocampus. CONCLUSION: The easier and quicker method of "slice counting" may be used to determine resection length expressed in "percent of total length", giving reliable values for resection length but causing less volumetric work. PMID- 18425624 TI - Vertical distribution and environmental significance of PAHs in soil profiles in Beijing, China. AB - Vertical distribution of both the concentration and composition of polycyclic aromatic hydrocarbons (PAHs) in ten profiles in Beijing has been investigated. The results showed that PAH concentrations and compositions in topsoil from different sampling sites were different. PAH concentrations were much higher in topsoil of the investigated urban area, industrial region, and paddy field with wastewater irrigation than in other areas. Moreover, PAH concentrations in topsoil were much higher than those at greater depth, where the concentrations were relatively consistent in most soil profiles. The fingerprints of PAHs in the samples from topsoil (0-30 cm) in the same profiles were similar and were obviously different from those at greater depth, suggesting that PAH sources were consistent in topsoil samples and were discriminating between topsoil and deeper soils. PAHs in topsoil mainly arose from mixed sources of combustion of liquid fuel, coal, and/or wood, as well as wastewater irrigation, while those at greater depth were derived from soil genesis and the process of soil formation. PMID- 18425623 TI - Use of the TIMI frame count in the assessment of coronary artery blood flow and microvascular function over the past 15 years. AB - Since its introduction, the TIMI frame count method has contributed to the understanding of the pathophysiology of coronary artery disease. In this article, the evolution of the TFC method and its applicability in the assessment of various therapeutic modalities are described. PMID- 18425625 TI - Evaluating the biodegradation of aromatic hydrocarbons by monitoring of several functional genes. AB - Various microbial activities determine the effectiveness of bioremediation processes. In this work, we evaluated the feasibility of gene array hybridization for monitoring the efficiency of biodegradation processes. Biodegradation of 14C labelled naphthalene and toluene by the aromatic hydrocarbon-degrading Pseudomonas putida F1, P. putida mt-2 and P. putida G7 was followed in mixed liquid culture microcosm by a preliminary, nylon membrane-based gene array. In the beginning of the study, toluene was degraded rapidly and increased amount of toluene degradation genes was detected by the preliminary gene array developed for the study. After toluene was degraded, naphthalene mineralization started and the amount of naphthalene degradation genes increased as biodegradation proceeded. The amount of toluene degradation genes decreased towards the end of the study. The hybridization signal intensities determined by preliminary gene array were in good agreement with mineralization of naphthalene and toluene and with the amount of naphthalene dioxygenase and toluene dioxygenase genes quantified by dot blot hybridization. The clear correlation between the results obtained by the preliminary array and the biodegradation process suggests that gene array methods can be considered as a promising tool for monitoring the efficiency of biodegradation processes. PMID- 18425626 TI - Overproduction and purification ofEscherichia coli tRNA(Leu). AB - Chemically synthesized genes encodingEscherichia coli tRNA (1) (Leu) and tRNA (2) (Leu) were ligated into the plasmid pTrc99B. then transformed intoEscherichia coli MT102, respectively. The positive transformants, named MT-Leu1 and MT-Leu2, were confirmed by DNA sequencing, and the conditions of cultivation for the two transformants were optimized. As a result, leucinc accepting activity of their total tRNA reached 810 and 560 pmol/A(260), respectively: the content of tRNA (1) (Leu) was 50% of total tRNA from MT-Leu1, while that of tRNA (2) (Leu) was 30% of total tRNA from MT-Leu2. Both tRNA(Leu)s from their rotal tRNs were fractionated to 1 600 pmol/A(260) after DEAE-Sepharose and BD-cellulose column chromatography. The accurate kinetic constants of aminoacylation of the two isoacceptors of tRNA(Leu) catalyzed by leucyl-tRNA synthetase were determined. PMID- 18425627 TI - Effects of G protein and cGMP on phytochrome-mediated amaranthin synthesis inAmaranthus caudatus seedlings. AB - The effects of G protein and cGMP on phytochrome-mediated amaranthin biosynthesis inAmaranthus caudatus seedlings were studied. It was shown that G protein agonist cholera toxin induced amarathin synthesis in darkness, whereas G protein antagonist pertussis toxin inhibited red light-induced amaranthin synthesis. Amaranthin synthesis was also induced by exogenous cGMP, while the amaranthin biosynthesis induced by cholera toxin, red light and exogenous cGMP was inhibited by genistein. L Y-83583, an inhibitor of guanylyl cyclase, inhibited the amarenthin synthesis induced both by red light and cholera toxin, while it was not able to inhibit the amaranthin synthesis induced by exogenous cGMP. These results suggest that G protein, guanylyl cyclase and cGMP were the candidates in phytochrone signal transduction chain for red light-induced amaranthin biosynthesis and the red light signal transduction chain might be as follows: red light --> phytochrome --> G protein --> guanylyl cyclase --> cGMP. PMID- 18425628 TI - Structural study on hen egg-white lysozyme crystals grown in gravity and microgravity. AB - Microgravity in space offers an ideal environment for growing protein crystals, where there is an absence of density-driven convection and sedimentation. Using domestic apparatus of protein crystalizetion, two space experiments of protein crystal growth have been carried out on the Chinese re-entry satellite, and good quality hen egg-white lysozyme crystals were obtained. To study the effect of microgravity on the prorein crystal structure, three structures of lysozyme crystals grown in gravity and microgravity were determined at high resolution. PMID- 18425629 TI - Activation and conformational changes of adenylate kinase in urea solution. AB - The activation and inactivation of adenylate kinase during deneturation in urea are compared with changes in UV absorbance at 287 nm. CD spectrum change at 222 nm, fluorescence intensity of ANS biding and small angle of X-ray scattering. At 1 mol/L, of urea the enzyme is activated 1.5-fold companied with a subtie decressing of its second structure, whereas its tertiary structure is fairly resistant to denaturation. By comparing the studies of the crystal structure and the mechanism of the catalysis of adenylste kinase, the activation is believed to result the effect that low concentration of urea increases the flexibility of the active site of the enzyme. This suggestion was confirmed by the results of the fluorescence intensity changes of ANS binding to adenylate kinase versus the concentration of urea. PMID- 18425630 TI - Presence, isolation and characterization of yolk DNA from chicken eggs. AB - Yolk DNA was detected with immunocytochemistry in fertilized chicken eggs. Yolk spheres were purifield with Ficoll-400 density gradient centrifugation, followed by extraction of DNA therein. Yolk DNA is sensitive to various restriction endonucleases. but shows no obvious bands, demonstrating its sequence heterogeneity. There is only a little, if any, methylated CpG in yolk DNA. judgmg from MspI and HpII digestion. Southern blot indicated that the sequence of yolk DNA accounts for a small portion of that of genomic DNA. Its uniqueness suggests that yolk DNA may play a crucial role in early development of chickens. PMID- 18425631 TI - Structural analysis of desheptapeptide(B24-B30) insulin by molecular replacement. AB - Desheptapeptide (B24-B30) insulin (DHPI), a virtually inactive insulin analog, has been crystallized in space group P2, 2, 2, with two DHPI molecules in an asymmetric unit. The orientatin and positions of the molecules were determind by molecular replacement. and a structural model was built at 0.3 nm resolution. The current model shows that the two DMHI monomers are related by a non crystallographic 2-fold axis, nearly parallel to the crystallographicc axis. This structural feature complicated the determination of the orientation of the local 2-fold axis, which was later confirmed by analysing the diffraction data of DHPI crystals. PMID- 18425632 TI - Two-dimensional crystallization and preliminary structure analysis of LHC-II from cucumber and spinach. AB - Large and well-ordered two-dimensional (2D) crystals of the light-harvesting chlorophyll a/b protein complexes (LHC-II) from cucumber and spinach chloroplasts were produced by the so-called batch method. The twodimensional structures of these crystals were examined at about 1.5 nm resolution by electron microscopy and image processing. The projection maps showed that there were similar, but not identical, structure features between two different LHC-II complexes. A cmparison between 2D crystal formations of the two different LHC-II complexes was done and some factors affecting 2D crystallization of the membrane proteins were analyzed. The relations of the structures of the LHC-II complexes to their polypeptide components and Chl a/b ratio were also discussed. PMID- 18425633 TI - Inheritance and mapping of embryo sac abortion in hybrid between Indica and Japonica rice (Oryza sativa L.). AB - A doubled haploid (DH) population derived from anther culture of F(1)'s between an Indica var. Zhai-Ye-Qing 8 and a Japonica var. Jing-Xi17 as well as two backcross populations derived from this DH population were used to investigate inheritance of the embryo sac abortion at early megagametogenesis occurring in Indica/Japonica rice crosses. Two major loci, dominant and complementaryesa-1 andesa-2, located on chromosomes 6 and 12 respectively, were detected. Genetic analysis indicated that embryo sac fertility is mainly regulated by the gametophytic genotype at these two loci. PMID- 18425634 TI - Mechanism of selective stabilization of extrinsic polypeptides in PSII particles by glycinebetaine. AB - The effect of glycinebetaine in selectively stabilizing the PSII extrinsic polypeptides was studied with PSII particles treated with different methods. It wan shown that glycinebetaine markedly stabilized the PSII extrinsic polypeptides when the partictes were treated with 0.8 mol/L Tris (pH8.0) or high concentrations of NaCI. The stabilizing effect was less pronounced when the PSII particles were treated by heat shok or trichlorocetate (TCA). The capability of halogenated acetates to release extrinsic polypeptides in PSII particles was found to follow the order of decreasing molecular hydrophobicities: trichloroacetate (TCA) > dichloroacetate (DCA) > monoiodoacetate (MIA) > monobromoacetate (MBA) > monochloroacetate (MCA). All these results imply that glycinebtaine is effective in stabilizing biopolymer structure against the action of electrolytes, while it is ineffective in protecting extrinsic polypeptides in PSII particles fm dissociation induced by halogenated acetates or heat treatments which disturbed the hydmphobic interactions within the polypeptides. PMID- 18425635 TI - EMS-induced mutant frequency and spectrum in bone marrow of D6-2 transgenic mice. AB - EMS-induced mutant frequency and mutation spectrum as well as background mutant frequency have been characterized fur bone marrow of the D6-2 transgenic mice. ThelacI genes carried on pSPORT1 vectors were recovered from the treated or untreated mouse genomic DNA by excision and circularization, and analyzedin vitro for mutations that occurred in the mouse bone marrow, lacI(-) mutants were positively selected with the M9/L media. The 6 lacI(-) mutants were identified out of 11 935 vectors recovered from genomic DNA of the treated mice (mutant frequency was 50 x 10(5)), while no mutant was found in 11 649 vectors Imm untreated mice (the background mutant frequency wan lower than 8.6 x 10(-5)). Two regions oflacI for each mutant, in which the majority of sensitive sites for inactivation of thelacI gene product have been located, were sequenced and 16 mutation events were identified. The predominant mutations (14/16 or 87.5%) were base substitutions, whereas the remaining 2 mutations were single base deletions (12.5%). Of these base substitutions, transversions made up 9/14 or 64%, and transitions cornprised 5/14 or 36%, These findings were markedly different from the spontaneous spectra characterized by using Big-Blue system, as well as from the EMS-induced mutation spectra obtained within vitro assay systems, where the EMS-induced predominant mutations are CG --> AT transitions. In addition, 45% of mutations analyzed occurred at CpG dinucleotides, which was in accordance with previous studies with other systems. These data show that: (i) the D6-2 transgenic mouse lineage is a suitable mdel for studying mutagenesisin vivo; (ii) a fundamental difference in mutagenesis for EMS betweenin nitro andin vivo assay systems may exist, but more extensive sequence analyses are required to determine the possible differences in mutation spectra. PMID- 18425636 TI - Epistasis plays an important role as genetic basis of heterosis in rice. AB - The genetic basis of heterosis in rice was studied in a cross between Zhenshan 97 and Minghui 63, the parents of the best hybrid widely used in rice production in China. Field data for yield and yield components traits were collected over two years from 240 F(2:3) families of this cross planted in replicated field trials. These families were assayed with 151 marker loci that were polymorphic between the parents and a linkage map was constructed with Mapmaker analysis. A total of 32 quantitative trait loci (QTLs) were identified for yield and the three component traits: 12 QTLs were detected in both years and the remaining 20 QTLs were observed in only one year. A search for the entire genome by using two-way analyses of variance with all possible two-locus combinations detected a very large number of significant digenic interactions involving both the QTLs resolved by single-locus analysis and loci that did not show significant effects by single locus analysis. Many of the interactions were simultaneously detected in both years. Partitioning of the epistatic interactions recovered all three types of interactions, i. e. additive by additive, additive by dominance and dominance by dominance, for all the traits. Some of the epistatic interactions even showed pleiotropic effects by simultaneously affecting two or more traits. The results clearly indicate that epistasis plays a significant role in the inheritance of yield traits as well as in the genetic basis of heterosis. PMID- 18425637 TI - NifL, an antagonistic regulator of NifA interacting with NifA. AB - Nitrogen fixation (nif) genes of diazotrophic enteric bacteria,Enterobacter cloacae orKlebsiella pneumoniae, are regulated bynif LA operon, in which thenif A product, NifA positively regulatesnif gene transcription, whereas the nifL product NifL represses it under oxygen or in excess of fixed nitrogen. Two-hybrid system was used to detect the possible interaction between NifA and NifL. The preliminary results illustrate that NifL does interact with NifA. The interaction between NifL and NtrC has also been shown. PMID- 18425638 TI - Agonist-induced down-regulation of alpha(1B)-adrenergic receptor in HEK293 cells transfected with alpha(1B) cDNA. AB - HEK293 cells stably expressing hamster alpha(1B)-adrenergic receptor (alpha(1B) AR) were used to observe the effect of nonepinephrine (NE) on alpha(1B)-AR gene expression. Radioligand binding assys and RNase protection assays were used to determine alpha(1B)-AR number and the mRNA level, respectively. Exposure (2-24 h) of HEK293 cella to NE (10 mumol) caused a decrease in alpha(1B)-AR mRNA with maximum change found at the 4th hour. and in alpha(1B)-AR density at the 24th hour. NE-induced decrease in alpha(1B)-AR mRNA was inhibited by protein kinase C (PKC) inhibitor calphostin C (0.1. mumol) and mimicked by PKC activator PMA (1 mumol). Nuclear run-off transcription assay showed that treatment of the cells with NE (10 mumol) exerted no effect on the transcription rate of alpha(1B)-AR, After the synthesis of new RNAs was inhibited by actinomycin D, NE muld not accelerate the degradation of alpha(1B)-AR mRNA. The results suggested that in the HEK293 cells NE muld induce the down-regulation of alpha(1B)-AR, and the effects were mediated by PKC pathway. NE could not alter the transcription rate of alpha(1B)-AR mRNA, but it might induce the synthesis of some factors and indirectly accelerate the degradation. PMID- 18425639 TI - Cloning and high-level expression of plasminogen activator inhibitor-1 cDNA derived from human glomerular mesangial cells. AB - Translation reading frame of human plasminogen activator inhibitor-1 (PAI-1) cDNA was amplified from total RNA extracted from glomerular mesangial cells by using reverse-transcription polymerase chain reaction (RT-PCR) and inserted into plasmid pUC19 in a sense orientation. Sequencing results revealed that PAI-1 cDNA had new initial codon ATG, but no signal peptide sequences, and translation reading frame was in agreement with PAI-1 cDNA derived from human endothelial cells. PAI-1 cDNA determined by sequencing was inserted into prokaryotic expression plasmid pBV220, and recombinant plasmid pBV220-PAI-l which had high level expression inEscherichia coli was obtained. Recombinant PAI-1 protein attained to approximately 45 % of total bacterial proteins. Western blotting showed that there was a specific band in the region of 43.0 ku. Latent recombinant PAI-1 with 97 % purity was obtained from inclusion bodies after denaturation, renaturation and purification by FPLC. Recombinant PAI-1 activated by treatment with 4.0 mol/L guanidinium-hydrochloride had significant inhibition activity to u-PA and shared bioactivity in common with natural PAI-1. PMID- 18425640 TI - Immune activation of erythroleukemia cells induced by interleukin 12. AB - To investigate the antitumor activity of IL-12, the induction of differentiation of IL-12 was observed using erythroleukemia cells (FBL-3) as model. After incubation with 200 U/mL IL-12 for 48 h, DNA synthesis of FBL-3 cells in S phase decreased significantly; the expression of CD14 which is the specific marker of monocyte increased, the rate of NBT(+) cells was apparently higher than that of the untreated FBL-3 cells. After treating FBL-3 cells with IL-12 for 72 h, the expression of 33D1 and NLDC145 which are the specific markers of dendritic cells increased markedly, the surface molecules such as MHC-11, B7-1, B7-2, and VCAM-1 were up-regulated; morphological observation showed two kinds of cells: some cells had a ruffled surface and plentiful lysosome; the others had many dendritic projections on the surface, and contained numerous mitochondria. Functionally, the IL-12-treated FBL-3 cells could apparently stimulate the proliferation of allogeneic and autologous T lymphocytes, and improve the specific cytotoxic activity of CTL on FBL-3 cells. These results indicated that erythroleukemia cells were induced by IL12 to differentiate into the monocytes and dendritic cells, then exhibited the antigen-presenting function. The data outline a new mechanism for IL-12 to treat leukemia. PMID- 18425641 TI - RecombinantPichia pastoris overexpressing bioactive phytase. AB - Phytase genephyA2, whose signal peptide encoding sequence and intron sequence had been removed, was modified. The Arg-encoding codons CGG and CAG inphyA2 were mutated into synonymous codon AGA. The modifiedphyA2 was fused behind a-factor signal sequence under the control ofAOX1 promoter in plasmid pPIC9, then introduced into the hostPichia pastoris by electroporation. The results of Southern blotting analysis and Northem blotting analysis demonstrated that thephyA2 gene had integrated into the genome ofP. pastoris and transcribed. The result of SDS-PAGE of the phytase expressed by P.pastoris showed that the modifiedphyA2 had been overexpressed and secreted. The concentration of the phytase expressed by P.pastoris with modifiedphyA2 exceeded 15 000 U/mL, which had a 3 000-fold increase over that of originAspergillus niger 963 and was 37 times higher than that of recombinantP. pastoris with non-modifiedphyA2. PMID- 18425642 TI - Certification of standard reference materials containing bitter orange. AB - A suite of three dietary supplement standard reference materials (SRMs) containing bitter orange has been developed, and the levels of five alkaloids and caffeine have been measured by multiple analytical methods. Synephrine, octopamine, tyramine, N-methyltyramine, hordenine, total alkaloids, and caffeine were determined by as many as six analytical methods, with measurements performed at the National Institute of Standards and Technology and at two collaborating laboratories. The methods offer substantial independence, with two types of extractions, two separation methods, and four detection methods. Excellent agreement was obtained among the measurements, with data reproducibility for most methods and analytes better than 5% relative standard deviation. The bitter orange-containing dietary supplement SRMs are intended primarily for use as measurement controls and for use in the development and validation of analytical methods. PMID- 18425643 TI - Electrically detected displacement assay (EDDA): a practical approach to nucleic acid testing in clinical or medical diagnosis. AB - This paper introduces the electrically detected displacement assay (EDDA), a electrical biosensor detection principle for applications in medical and clinical diagnosis, and compares the method to currently available microarray technologies in this field. The sensor can be integrated into automated systems of routine diagnosis, but may also be used as a sensor that is directly applied to the polymerase chain reaction (PCR) reaction vessel to detect unlabeled target amplicons within a few minutes. Major aspects of sensor assembly like immobilization procedure, accessibility of the capture probes, and prevention from nonspecific target adsorption, that are a prerequisite for a robust and reliable performance of the sensor, are demonstrated. Additionally, exemplary results from a human papillomavirus assay are presented. PMID- 18425644 TI - Determination of fluoroquinolone antibiotics in hospital and municipal wastewaters in Coimbra by liquid chromatography with a monolithic column and fluorescence detection. AB - The main goal of this work was determination of residues of the antibiotics ofloxacin (OFLO), norfloxacin (NOR), ciprofloxacin (CIPRO), and enrofloxacin (ENRO) in wastewater samples. The samples, after acidification to pH 4.5 and addition of EDTA, were extracted on an anion-exchange cartridge in tandem with an Oasis HLB cartridge. The LC-FD method, developed in previous studies, was based on application of a monolithic C(18) column. The limit of quantification (LOQ) of the method was 250 ng L(-1) for OFLO, 25 ng L(-1) for NOR and CIPRO, and 50 ng L( 1) for ENRO. Mean recovery ranged between 75 and 121% for OFLO, NOR, CIPRO, and ENRO. A total of 14 wastewater samples were analyzed; these were collected from four hospitals and from influent and effluent from a wastewater-treatment plant in Coimbra, Portugal, during spring and autumn. CIPRO was present in all the samples, NOR was detected second most often, followed by OFLO. ENRO was found at concentrations under the LOQ in five hospital samples, and the highest level was found in influent from the WWTP. PMID- 18425647 TI - Introduction to the special issue: innovative approaches to health behavior change. PMID- 18425645 TI - Multi-residue analysis of traces of pesticides and antibiotics in honey by HPLC MS-MS. AB - The aim of this work was to develop an analytical method for simultaneous assay of residues of two families of antibiotics, and three pesticides, in honey. The assays involved a mixture of five tetracyclines, four sulfamides, and the pesticides coumaphos, carbendazim, and amitraz (two metabolites). All the compounds were extracted from honey and pre-concentrated by optimised solid-phase extraction (SPE). Analysis was by high-performance liquid chromatography-mass spectrometry-mass spectrometry (HPLC-MS-MS) using a triple-quadrupole spectrometer in multiple reaction monitoring (MRM) mode in order to identify and quantify the compounds present (Sheth et al J Agric Food Chem 38:1125-1130, 1990). During development of the analytical method a strong matrix effect was found that depended on the floral origin of the honey. This led to the development of a standard additions method to quantify the contaminants sought. PMID- 18425648 TI - Shared medical decision-making: a new paradigm for behavioral medicine--1997 presidential address. AB - Different conceptual models lead to different health care choices. The traditional biomedical model emphasizes identification of pathology (diagnosis) and remediation of these biological deficits (treatment). An alternative approach, known as the outcomes model, focuses attention on the outcomes of health care. Specifically, health care is regarded as effective only if it extends life or if it improves quality of life. Indices that combine life expectancy and life quality can be used to monitor the benefits of health care. According to the traditional model, medical care is effective if it improves a clinical indicator (i.e. reduces blood pressure, decreases tumor size, etc.). According to the outcomes model, treatments are not advocated unless they improve general outcomes. There are circumstances in which clinical indicators improve but general outcomes remain the same or get worse. Data on the detection and treatment of prostate cancer are used to illustrate how these models might lead to different treatment decisions. According to the traditional model, aggressive screening and treatment of prostate cancer should be advocated because more cases are detected early and more tumors are removed. According to the outcomes model, net quality-adjusted life may be reduced rather than enhanced with screening. Shared medical decision-making is an outgrowth of the outcomes model. Using these methods, patients and providers integrate the best scientific evidence on treatment efficacy with patient preferences for outcomes. Often shared decision making leads to reductions in the use of medical procedures. PMID- 18425649 TI - Reducing denial and sexual risk behaviors in college students: a comparison of a cognitive and a motivational approach. AB - This study compared two interventions designed to decrease denial of sexually transmitted disease (STD)/human immunodeficiency virus (HIV) risk, to increase purchase of and intent to use condoms, and to decrease sexual risk behaviors. One hundred and fifty sexually active male and female undergraduates were assessed at baseline for denial of STD/HIV risk, knowledge of sexual risk behaviors, and self reports of sexual behavior and were randomly assigned to either a motivational or cognitive intervention or a control condition. After the intervention, subjects were offered the opportunity to purchase condoms and were assessed for denial and intent to use condoms. Two months later, sexual risk behaviors and denial were measured. The motivational approach was most effective in reducing denial and in increasing intent to use condoms immediately following the intervention and in reducing sexual risk behaviors 2 months later. PMID- 18425650 TI - Comparison of five teacher actions to encourage children's new food acceptance. AB - How can teachers encourage children to accept new fruits and vegetables? A quasi experimental study with 64 preschool children (32 boys, 32 girls) compared the effectiveness of five teacher actions to encourage children's acceptance of four new fruits and vegetables presented during three preschool lunches. The five teacher actions included reward (special dessert), modeling, insisting children try one bite, choice-offering ("Do you want any of this?"), and a control condition of simple exposure.In factorial analyses of variance (two genders x five teacher actions), the five teacher actions produced differences in number of foods sampled (p<.001), number of meals during which foods were sampled (p<.004), and total number of bites (p<.002). Paired comparisons revealed that reward, insisting, and choice-offering were more effective than simple exposure to encourage number of foods, number of meals, and number of bites. Dessert reward and choice-offering were equally effective for all three measures of new food acceptance, but insisting produced fewer bites than did choice-offering. Under the present conditions, teacher modeling was ineffective compared to simple exposure. No gender differences were found in new food acceptance or in interactions with the five teacher actions to encourage new food acceptance. PMID- 18425651 TI - Monitoring styles in women at risk for cervical cancer: implications for the framing of health-relevant messages. AB - We explored the interaction effects of individual attentional style (high versus low monitoring) and the framing of informational messages on the responses of women undergoing diagnostic follow-up (colposcopy) for precancerous cervical lesions. Prior to the colposcopic procedure, patients (N=76) were randomly assigned to one of three preparatory conditions: (a) Loss-framed message, which emphasized the cost of nonadherence to screening recommendations; (b) Gain-framed message, which emphasized the benefit of adherence; and (c) Neutrally-framed message. It was hypothesized that low monitors (who are more positively biased about their health) would show a more adaptive pattern of response to loss-framed information than high monitors (who are more negatively biased about their health). The results of a series of hierarchical multiple regression analyses were consistent with this prediction. Low monitoring was associated with greater knowledge retention (beta=.61, p<.05) and less canceling/rescheduling of follow up appointments in the loss condition than in the neutral condition (beta=.82, p<.002). High monitoring, however, was associated with greater intrusive ideation when information was presented in the loss-oriented frame as compared to the neutral frame (beta=.99, p<.01). Knowledge retention and screening adherence were not affected by the framing manipulation. The differences between high versus low monitors as a function of loss or neutral frame suggest an interaction effect, wherein both the type of framing message and the individual's attentional style lead to distinctive cognitive-affective and behavioral patterns. The findings may have clinical implications for the tailoring of health messages to the individual's signature style. PMID- 18425652 TI - Calorie restriction is more effective for obesity treatment than dietary fat restriction. AB - Recent evidence suggests that dietary fat intake may be more strongly associated than total energy intake to the development and maintenance of obesity. The objective of this study was to determine whether dietary fat restriction would promote more favorable changes in body weight, body composition, resting metabolic rate (RMR), eating behavior, and dietary adherence than calorie restriction. Sixty-five obese women and 15 obese men were recruited and randomly assigned to calorie restriction (1000-1200 kcal/day) or fat restriction (22-26 g/day). Subjects participated in a 24-week behavioral weight loss program. Forty eight subjects completed assessments at all four time points-baseline and 6, 12, and 18 months. Weight loss was significantly greater in the low calorie (LC) group (-11.2 versus -6.1 kg, p<.001) and was maintained better at the 18-month assessment (-7.5 versus -1.8 kg, p< or =.001). There was a significantly greater decrease in percent body fat and RMR for the LC group with only percent body fat still significantly lower at 18 months. Subjects in the LC group consumed significantly fewer calories, less carbohydrates, and the same amount of fat as those in the low fat (LF) condition, however, calorie and carbohydrate intake were decreasing for LF subjects by the 12- and 18-month assessments. There were no long-term differences in most measured predictors of dietary adherence. Dietary fat restriction did not prove to be superior to calorie restriction, thus strengthening the public health message that calories do count. PMID- 18425653 TI - Physician-based physical activity counseling for middle-aged and older adults: a randomized trial. AB - Sedentary behavior among older adults increases risk for chronic diseases. Physicians in a primary care setting can play an important role in promoting physical activity adoption among their older patients. The Physically Active for Life (PAL) project was a randomized, controlled trial comparing the efficacy of brief physician-delivered physical activity counseling to usual care on self reported physical activity levels. The physical activity counseling was based on the Transtheoretical Model of Change and social learning theory. Twenty-four community-based primary care medical practices were recruited into the study; 12 were randomized to the Intervention condition and 12 to the Control condition. Physicians in the Intervention practices received training in the delivery of brief physical activity counseling. Subjects in the Intervention practices (n=181) received brief activity counseling matched to their stage of motivational readiness for physical activity, a patient manual, a follow-up appointment with their physician to discuss activity counseling, and newsletter mailings. Subjects in the Control practices (n=174) received standard care. Measures of motivational readiness for physical activity and the Physical Activity Scale for the Elderly (PASE) were administered to subjects in both conditions at baseline, 6 weeks following their initial appointment, and at 8 months. Results showed that at the 6-week follow-up, subjects in the Intervention condition were more likely to be in more advanced stages of motivational readiness for physical activity than subjects in the Control condition. This effect was not maintained at the 8 month follow-up and the intervention did not produce significant changes in PASE scores. Results suggest that more intensive, sustained interventions may be necessary to promote the adoption of physical activity among sedentary, middle aged, and older adults in primary care medical practices. PMID- 18425654 TI - Evaluation of a dental office tobacco cessation program: effects on smokeless tobacco use. AB - We describe a randomized trial designed to evaluate the effectiveness of a smokeless tobacco cessation intervention delivered by dental hygienists as part of a patient's regularly scheduled cleaning visit. Seventy-five practices were randomized to continue their usual care (n=25; 239 smokeless tobacco using patients enrolled) or to receive training to provide a tobacco cessation intervention (n=50; 394 smokeless tobacco using patients enrolled). Patient reports indicated that the training program was successful in getting hygienists to implement the intervention. The intervention produced a strong effect on sustained quitting for smokeless tobacco users but had no impact on secondary outcomes, including unsuccessful quit attempts, future intent to quit using smokeless tobacco, and change in readiness to quit using. Frequency of smokeless tobacco use and receipt of specific components of the intervention, including the video and written materials, predicted sustained cessation. Since this intervention was delivered by dental hygienists as part of a patient's regularly scheduled cleaning visit, it is easily disseminable. PMID- 18425655 TI - Do enhanced friendship networks and active coping mediate the effect of self-help groups on substance abuse? AB - Self-help groups are the most commonly sought source of help for substance abuse problems, but few studies have evaluated the mechanisms through which they exert their effects on members. The present project evaluates mediators of the effects of self-help groups in a sample of 2,337 male veterans who were treated for substance abuse. The majority of participants became involved in self-help groups after inpatient treatment, and this involvement predicted reduced substance use at 1-year follow-up. Both enhanced friendship networks and increased active coping responses appeared to mediate these effects. Implications for self-help groups and professional treatments are discussed. PMID- 18425656 TI - The thresholds of change model: an approach to analyzing stages of change data. AB - Stage models are prominent in research describing health behavior change. Since stage models often propose that different factors have varying influences on membership in the different stage, statistical methods that can estimate the thresholds that separate the stages and the relative value of variables in influencing these thresholds are useful. This article describes use of a "thresholds of change" model for analyzing the thresholds separating stages and specifically for examining the effects of explanatory variables on these thresholds using a generalization of an ordinal logistic (or probit) regression model. Data from a skin cancer prevention study (N=3,185) in which participants were grouped into three stages for sunscreen use (precontemplation, contemplation, and action) are used to illustrate the Thresholds of Change Model. For this example, two thresholds exist: a contemplation (between precontemplation and contemplation) and an action threshold (between contemplation and action). Variables examined include gender, skin type, perceived susceptibility to sunburn, worry about skin cancer, and sun protection self-efficacy. We examine models that assume that the effects of these variables are the same across thresholds, and then allow the effects of these variables to vary across thresholds. Results indicate that perceived susceptibility has an equal effect on both thresholds, but that worry and self-efficacy have differential effects: worry exerts a greater influence on the contemplation threshold, whereas self efficacy has a significantly stronger effect on the action threshold. Gender also has a stronger effect on the action threshold; males were less likely to be classified in the action stage than females. This analytic approach has broad applications to many types of stage data. PMID- 18425657 TI - When does cigarette fading increase the likelihood of future cessation? AB - OBJECTIVE: To determine which smokers report cigarette fading, how much they fade, when fading leads to quitting, and, if not, whether it can be maintained. METHODS: Subjects were 1,682 adult smokers interviewed by telephone in 1990 and 1992 as part of the California Tobacco Survey. Data from three timepoints in the same subjects were compared. At Time 1 (one year before the baseline survey), all respondents were daily smokers who recalled their average daily cigarette consumption retrospectively at baseline. At Time 2 (baseline survey), all respondents were current smokers who provided concurrent data on their average daily cigarette consumption. At Time 3 (follow-up), smoking status and current cigarette consumption among nonabstinent respondents were assessed. RESULTS: Nearly 18% of smokers reduced consumption between Times 1 and 2. The mean reduction was 13 cigarettes per day. Only moderate to heavy smokers who reduced consumption to below 15 cigarettes per day were more likely to be in cessation at Time 3 (24.9% versus 5.8%, respectively). The cessation rate for moderate to heavy smokers that became light smokers by baseline was similar to that for smokers who were already light smokers 1 year before baseline. Continuing smokers who reduced consumption between Times 1 and 2 maintained a mean reduction of 11.4 cigarettes per day. CONCLUSIONS: Cigarette fading increases cessation among moderate to heavy smokers who become light smokers. PMID- 18425658 TI - Relationships among patient hostility, anger expression, depression, and the working alliance in a work hardening program. AB - Hostility, anger expression, and depression may adversely affect the response of chronic pain patients to pain or functional restoration programs by hampering the development of good working alliances between patients and physical or occupational therapists. Measures of hostility, anger expression, and depression and the Working Alliance Inventory (WAI) were completed by 71 patients with chronic pain (stemming from work-related injuries) prior to starting a "work hardening" program. The physical or occupational therapist acting as any given patient's "program manager" completed the WAI, as well. Hostility and anger expression were correlated negatively only with patient ratings of the working alliance. Therapist ratings were predicted by the interaction of patient anger expression and depression, such that therapists reported their poorest alliances with patients who were both depressed and expressed anger. Results imply that hostility and the propensity to express anger may diminish a pain patient's capacity to foster a collaborative relationship with physical or occupational therapists, while therapists may readily become alienated from depressed and irritable patients. PMID- 18425659 TI - Is life more difficult on Mars or Venus? A meta-analytic review of sex differences in major and minor life events. AB - We conducted a meta-analysis of studies examining sex differences in reported levels of stress, considering the impact of: (a) the age and representativeness of sample participants, (b) whether life events were weighted or unweighted by participants for impact or severity, (c) the major versus minor nature of the stress, and (d) the life domain of the stressor. Overall, the meta-analysis of 119 studies including 83,559 participants found that females were exposed to more stress than were males (d=.123, r=.061). However, there was considerable heterogeneity among studies, with greater effect sizes associated with: (a) life events weighted by participants for impact, (b) adolescents compared to both younger and older samples, (c) major life stressors compared to minor stressors, and (d) interpersonal relationship stressors compared to work stressors. In none of the subgroup analyses did males experience considerably more stress than females. Evaluation of a subsample of 39 studies that examined gender differences in psychological symptoms revealed that females reported more symptoms of depression, anxiety, and psychosomatic problems (d=.282, r=.139) and that the sex difference in reports of psychological symptoms accounted for approximately 4% of the variance in the sex differences in reports of stress. Possible explanations for the observed patterning of effects are discussed, as are recommendations for further research. PMID- 18425660 TI - Effect of cholesterol-lowering diets on indices of depression and hostility. AB - Increased injury deaths have been reported among treatment groups in cholesterol lowering trials, leading to speculation that lipid lowering may result in behavioral disorders. We investigated this in 319 men enrolled in a 2-year trial of lipid lowering diets who completed measures of depression and hostility at entry and 24 months later. Mean Beck Depression Inventory (BDI) scores were lower after 24 months (3.8 versus 3.3,p<0.05) and Symptom Checklist 90-Revised (SCL-90) depression and hostility scores were unchanged. After adjustment for potential confounding, 24-month hostility and BDI scores were unrelated to lipid changes. A small inverse association of borderline statistical significance (B=0.034,p=0.08) was noted between 24-month SCL-90 depression scores and lipid changes. Lipid lowering diets had no significant adverse effect on psychological function and are consistent with current dietary recommendations. PMID- 18425661 TI - The relationship between coping and psychological well-being among people with osteoarthritis: a problem-specific approach. AB - We examined the strategies that people with osteoarthritis (OA) use to cope with illness-related problems in four areas: household activities, leisure activities, pain management, and social relationships. We also examined the relationship between the coping strategies participants reported using upon entry to the study (Time 1) and psychological well-being reported six months later. Three hundred people, aged 50 and over, with OA participated in the study. Data were collected via two mailed questionnaires, administered at six-month intervals. We found that two of the coping strategies examined, self-criticism and social withdrawal, were used more frequently for social relationship problems than for any of the other three types of problems. None of the other coping strategies (i.e. problem solving, cognitive restructuring, social support, emotional expression, problem avoidance, turning to religion, information seeking) were used differentially across problem areas. Multivariate analyses revealed that the coping strategies people used at Time 1 significantly predicted psychological well-being, as assessed by measures of positive affect, negative affect, and depressive symptoms, six months later. However, the specific strategies that predicted positive affect were different from those that predicted negative affect and depressive symptoms. Implications of these findings for future research on coping with chronic illness are discussed. PMID- 18425662 TI - Stress and reactivation of latent herpes simplex virus: a fusion of behavioral medicine and molecular biology. AB - Since 1978, the study of health and behavior has become a major focus of scientists in psychology, psychiatry, nursing, neuroscience, and in traditional medical science disciplines. Investigation of psychological or behavioral influences on biological systems has established that biobehavioral processes such as stress play an important role in disease processes. An excellent example of the interactions between stress and health outcomes is the reactivation of latent herpes simplex virus (HSV) leading to recurrent lesions. This article describes what is currently known about HSV latency and reactivation and considers some mechanisms by which stress-induced changes in the host's immune and nervous systems might allow for either the establishment or reactivation of latent viral infections. PMID- 18425663 TI - Social relationships, recovery from illness, and survival: a literature review. AB - Do medical patients with a high quantity or quality of social relationships have greater chances of recovery and survival than more isolated individuals? This article reviews longitudinal studies of social relationships and recovery published since the last major reviews of this field. Reports of 26 such projects were located, primarily in the areas of heart disease (13 studies) and breast cancer (7 studies). Being married (or socially supported in other ways) was generally associated with survival or freedom from recurrence in multiyear follow up studies of myocardial infarction (MI) and coronary artery disease patients, although social support produced negative or mixed results in studies of short term physical adaptation after Ml or bypass surgery. Studies relating marital status and other support variables to recurrence and survival in breast cancer patients also had mixed results. The small number of studies, and other limitations associated with them, suggest caution in drawing strong conclusions. PMID- 18425664 TI - Weight gain after childbirth: a women's health concern? AB - Changes in health care delivery call for an integrative science of women's health care. A neglected area of such a science is the interrelationship between childbearing and chronic health conditions, such as obesity. Recent increases in guidelines for gestational weight gain (GWG) have raised concerns that these guidelines may contribute inadvertently to later obesity in women. On average, existing research shows that pregnancy accounts for small gains in postpartal weight, especially if the effects of aging are considered. However, there is great variability in postpartal weight gain, and some women, especially minority women, are more vulnerable to excessive weight gains following childbirth. Although GWG is an important predictor of postpartal weight gain, psychosocial factors, such as personal health patterns and stress, also may potentially contribute to women's weight gain after childbirth. Further research is needed to determine the impact of women's psychosocial context on postpartal weight. PMID- 18425665 TI - Depression as a risk factor for cardiac events in established coronary heart disease: a review of possible mechanisms. AB - Depression is highly prevalent in patients with coronary heart disease (CHD) and is associated with excess medical morbidity and mortality. This article reviews the mechanisms which may mediate the relationship between depression and cardiac events in these patients. Research in this area is at an early stage of development. However, there is evidence that altered autonomic tone, smoking, hypertension, and, at least in older patients, poor adherence to the cardiac treatment regimen may be responsible for the adverse cardiac effects of depression. PMID- 18425667 TI - From human papillomavirus (HPV) to cervical cancer: psychosocial processes in infection, detection, and control. AB - Certain high-risk types of human papillomaviruses (HPVs), which are sexually transmitted persistent, and currently epidemic in the United States, are important etiologic agents in cervical cancer, constituting an acute health threat to women. Consequently, adherence to recommended Papanicolaou (Pap) screening and colposcopy regimens is crucial for early diagnosis and treatment which enhance survival dramatically. This article reviews research and theory on major psychosocial factors relevant to the transmission, onset, and course of this infection. The review focuses on cognitive-affective variables and processes (e.g. monitoring-blunting, anxiety, depression) that underlie--and potentially undermine--adherence to relevant health-protective behaviors. Identification of these processes should facilitate interventions to help women, particularly from low-income, inner city populations who are at highest risk, to adhere to essential follow-up regimens. PMID- 18425669 TI - Cigarette smoking among adult Latinos: the California Tobacco Baseline Survey. AB - This article presents probability estimates of smoking prevalence for the Latino adult population in California and examines differences in smoking prevalence and nicotine dependence between different levels of education and acculturation and between different ethnic and gender groups. Prevalence estimates are based on the California Baseline Tobacco Survey (CTS). The CTS is a Random Digit Dial Survey including N=70,997 non-Latino Whites and 28,000 Latino adults. Overall smoking prevalence among Latinos was estimated at 24.4% for men and 12.0% for women. Multiple logistic regression analyses suggest higher smoking prevalence and higher nicotine dependence for men than for women, for individuals who have not finished high school, for non-Latinos when compared to Latinos, and for Latinos of high level of acculturation when compared to Latinos of low level of acculturation. These estimates can serve as baselines to evaluate progress toward year 2000 health objectives. Findings stress the importance of taking into account socio-economic level when comparing Latinos and other ethnic groups on health behaviors. Moreover, findings suggest that interventions helping Latinos quit smoking are most needed for men with twelve years or less of formal education and stress the importance of programs to prevent smoking uptake, specifically among Latinos of low level of acculturation. PMID- 18425668 TI - Epinephrine and cortisol responses to work: a test of the models of Frankenhaeuser and Karasek. AB - Both Frankenhaeuser and Karasek have put forward models describing how job demand and control influence epinephrine and cortisol levels. These models were tested in a sample of 53 women and 51 men in a variety of occupations. They were studied over one rest day and two working days. Subjects reported their perceived demand and control and their mood on each day, as well as providing urine for assessment of urinary excretion rates of epinephrine and cortisol. In men, but not women, epinephrine levels were higher on the working days than on the rest day, and demand was found to covary positively with epinephrine, supporting Frankenhaeuser's model with respect to epinephrine variation in men. However, cortisol levels were not elevated on working days compared to the rest day, and no relationship between job control and cortisol was seen which is in contradiction of Frankenhaeuser's model with respect to cortisol variation. There was some suggestion that demand was most strongly associated with elevated epinephrine in men when job control was low in accordance with Karasek's model, but there was no evidence for such an effect with respect to cortisol. PMID- 18425670 TI - Defensive hostility, gender and cardiovascular levels and responses to stress. AB - Prior studies have demonstrated inconsistent relationships between the trait of hostility and cardiovascular responses to stress. To examine the hypothesis that only a subset of hostile subjects demonstrates greater cardiovascular responses to stress, we assessed relationships among hostility, defensiveness, and cardiovascular responses to stress in 33 healthy men and 34 healthy women. Stressors used were math and speech tasks. Median splits on Cook-medley Hostility Inventory (Ho) and defensiveness [Marlowe-Crowne Social Desirability scale (MC)] classified subjects into four groups: (a) Defensive Hostile (DH--high Ho and MC), (b) Low Hostile (LH--low Ho and MC), (c) High Hostile (HH--high Ho, low MC), and (d) Defensive (Def--low Ho, high MC). Results indicate that Defensive Hostility is differentially related to cardiovascular levels in men and women. DH men exhibited greater systolic blood pressure (SBP) levels than LH, Def, and HH men. Analyses of diastolic blood pressures (DBPs) suggested a trend that LH and DH men demonstrated greater DBP levels than Def and HH men. In contrast, DH women were indistinguishable from HH and Def women with respect to blood pressure, and LH women demonstrated the lowest SBP and DBP levels. There were no personality differences in cardiovascular change scores to stress. Gender differences for affect were observed which may mediate the cardiovascular responses. These data suggest that the personality trait of Defensive Hostility may provide significant associations with blood pressure levels and coronary disease associations with blood pressure levels and coronary disease in studies that do not find associations using hostility alone. PMID- 18425671 TI - Effect of stress on opioid-seeking behavior: evidence from studies with rats. AB - Studies concerned with the relation between exposure to stress and the behavioral effects of opioid agonists in animal models of drug use are reviewed. These studies, which primarily utilized male rats, indicate that under certain conditions short-term mild stressors increase self-administration of opioid drugs and reinstate herein-seeking behavior following a drug-free period. On the other hand, there is evidence that long-term chronic inescapable stressors and severe acute stressors reduce the reinforcing effects of morphine as measured by a conditioned place preference procedure and decrease the behavioral effects of other positive reinforcers. The results of the studies reviewed suggest that stressors are important modulators of opioid-taking behavior, especially during drug-free periods. The implications of these findings to the understanding of the neurobiology of relapse to opioid-seeking behavior and for strategies for medication development to prevent relapse to heroin are discussed. PMID- 18425672 TI - Comparison of adjustment, activity, and tangible social support in men and women patients and their spouses during the six months post-myocardial infarction. AB - A longitudinal study was conducted to investigate gender differences in adaptation and activity among survivors of acute myocardial infarction (MI) and their spouses during the six months post-hospital discharge. Male and female married survivors of MI, matched on age, disease severity, and socioeconomic status, and their spouses responded to measures about functional impairment, psychological distress, and level of involvement in household and other activities prior to and 4, 10, 16, and 22 weeks after the MI. Both patients and spouses were distressed by the MI, but the distress lasted longer for spouses. Male spouses increased traditional domestic activities (e.g. cooking, laundry) in the weeks shortly after their wives' heart attack, and patient wives decreased domestic activities compared to prior to the MI. However, wives still did as much as their husbands. Women recovering from MI may carry a heavier burden of household responsibilities and activities than their male counterparts. This burden, if premature, may pose a threat for reinfarction during the early stages of rehabilitation. PMID- 18425673 TI - Changes over time in weight concerns among women smokers engaged in the cessation process. AB - Weight concerns have been reported by women smokers to be barriers to initial cessation and to sustained abstinence. This article examines the temporal patterns of weight concerns and self-efficacy for cessation among three groups of women smokers: non-quitters, short-term quitters, and long-term quitters. Subjects were 417 women aged 20-64 who had participated in a randomized smoking cessation intervention trial. Over the twelve-month follow-up, long-term quitters reported greater increases in weight gain, pain and worry related to weight, dieting behaviors, and self-efficacy for maintaining cessation in eating-related situations compared to non-quitters and short-term quitters. In multivariate analyses, increases in pain and worry about weight and in self-efficacy in eating related situations were significantly associated with sustained abstinence. Cessation-specific weight concerns and dieting were not associated with sustained abstinence. Implications of these results for intervention design are discussed. PMID- 18425674 TI - Methodological issues in weight cycling. AB - Recent studies have suggested that weight changes may be related to disease risk independent of weight status. A critical step in testing this assertion is the measurement of weight change and so-called "weight cycling". However intuitive the concept of weight cycling may appear, research in this area is hampered by complex methodological issues. This article discusses various measures of nominal weight cycling, including the standard deviation, coefficient of variation, regression techniques, and cycles. A cycle is a sequence of a gain followed by a loss or vice versa. The various measures are compared in seven hypothetical cases created to illustrate their strengths and weaknesses. Superior performance of the cycles measure over the coefficient of variation, number of fluctuations, and simple regression methods is argued. The linkage of the cycles measure with the statistical theory of runs also provides a basis for testing the significance of weight fluctuations or other variables that may cycle, such as blood lipids, etc. The cycles measure and runs test provide a viable definition for identifying weight cycling and a tool for evaluating the critical amount of weight gained and/or lost in relationship to risk. PMID- 18425676 TI - Analysis of the progression of intraductal proliferative lesions in the breast by PCR-based clonal assay. AB - PURPOSE: To analyze the progression in patients with a morphological diagnosis of intraductal proliferative lesions by PCR-based clonal assay. MATERIALS AND METHODS: An X-chromosome inactivation assay was applied to explore clonal relationships in human intraductal proliferative lesions of the breast. Four groups samples, including 40 cases of usual ductal hyperplasia (UDH), 40 cases of atypical ductal hyperplasia (ADH), 29 cases of flat epithelia atypia (FEA), and 40 cases of ductal carcinoma in situ (DCIS) were selected for analysis. Thirty specimens of normal breast tissue were used as a control group. Microdissection was performed to collect the tissue samples for extraction of genomic DNA from paraffin-embedded tissues. The DNA was subjected to PCR amplification of the CAG repeats in androgen receptor (AR) gene exon I with and without prior digestion of methylation-sensitive restriction enzyme HhaI. Gel electrophoresis was used to detect the clonal nature of these four groups samples. RESULTS: The clonal analysis confirmed monoclonality in all informative samples of DCIS cells. Normal tissues and the majority (97.1%) of UDH were shown to be polyclonal. Monoclonality was revealed in 20/39 (51.3%) cases of ADH. Among 26 cases of FEA, 20 were shown to be polyclonal, while six displayed monoclonal alterations which accounted for 23.1%. CONCLUSION: These findings reinforce recent suggestions that clonal analysis with AR gene polymerase chain reaction may be used to define the genetic relationships among the human tumor and the breast intraductal proliferative lesions. Furthermore, our observations demonstrate nearly a half ADH and the smaller part of FEA have clonal alterations, which may be neoplastic lesions. This method would shed light on genetic abnormalities that play a role in early tumorigenesis of the breast, and thus might be an adjunct in predicting the probability of breast tumor occurrence and in guiding the management of these cases. PMID- 18425675 TI - Integrating individual and public health perspectives for treatment of tobacco dependence under managed health care: a combined stepped-care and matching model. AB - There is an increasing momentum to integrate prevention into mainstream health care. Three decades of research on tobacco dependence can provide insights into the conceptual, clinical, economic, and service delivery challenges to such an integration. Biological sciences, cognitive-behavioral, clinical treatment outcome, and public health arenas are selectively reviewed. The key conceptual issues are explored relevant to the optimal delivery of quality smoking cessation treatments for the general population of adult smokers at reasonable cost. A comprehensive model for adult smoking cessation treatment is developed. The model consists of an overarching public health approach, focusing on enhancing motivational level from low motivation to quit to high motivation. A common outcome metric of overall impact is proposed to facilitate comparisons between clinical and public health interventions. Smokers are then assessed and triaged into one of three treatment steps of minimal, moderate, and maximal intensity and cost. Smoker individual differences at both the population and individual level are also taken into account as part of a tailoring or matching strategy within and across the stepped interventions. Smoker profiles include sociocultural, nicotine dependence, and comorbidity factors. The result is a hybrid stepped-care matching model. The model serves to illustrate some of the needs and challenges facing future tobacco dependence research and practice. Comparisons are made between tobacco control and other preventive medicine practices in terms of cost per quality adjusted life-year saved. The barriers and opportunities under managed care are explored. The conceptual principles identified here could be used as a guidepost for integrating other preventive medicine programs into the evolving managed health care system. PMID- 18425678 TI - The clinical significance of sentinel lymph node micrometastasis in breast cancer. AB - INTRODUCTION: The advent of sentinel lymph node biopsy (SLNB) and improvements in histopathological and molecular analysis have increased the rate at which micrometastases (MM) are identified. However, their significance has been the subject of much debate. In this article we review the literature concerning axillary lymph node (ALN) MM, with particular reference to SLNB. The controversies regarding histopathological assessment, clinical relevance and management implications are discussed. METHODS: Literature review facilitated by Medline and PubMed databases. RESULTS: Published studies have reported divergent results regarding the significance and implications of ALN MM in general and sentinel lymph node (SLN) MM in particular. Some studies demonstrate no associations, whilst others have found these to be indicators of poor prognosis, associated with non-SLN involvement, in addition to local and distant failure. Absolute consensus regarding the optimal analytical technique for SLNs has yet to be reached, particularly concerning immunohistochemical (IHC) techniques targeting cytokeratins and the utility of contemporary molecular analysis. CONCLUSION: SLN MM are likely to represent an incremental detriment to prognosis and increased risk of non-SLN involvement, despite only modest up-staging within current classification systems. In the absence of level-1 guidance concerning the management of women with SLN MM, each case requires discussion with regard to other tumour and patient related factors in the context of the multidisciplinary team. Randomized studies are required to evaluate the prognostic significance and optimal management of each category of tumour burden within the SLN. The identification of MM remains highly dependent on the analytical technique employed and there exists potential for stage migration and impact on management decisions. PMID- 18425677 TI - Radiofrequency thermal ablation of breast tumors combined with intralesional administration of IL-7 and IL-15 augments anti-tumor immune responses and inhibits tumor development and metastasis. AB - Tumor development or recurrence is always a matter of concern following radiofrequency thermal ablation (RFA) of tumors. To determine whether combining RFA with immunologically active cytokines might induce tumor-specific immune responses against mammary carcinoma and inhibit tumor development or metastasis, we evaluated intralesional injection of IL-7 and IL-15 in RFA-treated murine tumors. We used two different breast carcinoma models: neu-overexpressing mouse mammary carcinoma (MMC) in FVBN202 transgenic mouse and 4T1 tumors in Balb/c mouse. MMC tend to relapse even in the presence of neu-specific immune responses, and 4T1 is a weakly immunogenic, aggressive and highly metastatic transplantable tumor. In vivo growth of both of these tumors is also associated with increased numbers of CD11b+Gr1+ myeloid-derived suppressor cells (MDSC). We showed for the first time that unlike RFA alone, RFA combined with the administration of intralesional IL-7 and IL-15 (after RFA), induced immune responses to tumors, inhibited tumor development and lung metastasis, and reduced MDSC. PMID- 18425680 TI - Some properties of Streptococcus thermophilus bacteriophages. AB - The morphology, host range, structural proteins and serological properties of Streptococcus thermophilus phages isolated from Finnish cheese plants were investigated. The results show that all the nine phages belong morphologically to Ackermann's group B1. The host-phage reactions and plating efficiency justify the division of these phages into four specificity groups. Most of the phages showed an absolute host specificity as to their plating efficiency but were not strictly specific in the adsorption to different hosts. The electrophoretic profiles of the structural proteins appeared nearly identical. Ten to eleven well separated proteins could be detected. The antiserum raised against one of the phages contained antibodies with different neutralization capacity depending on the phage. Using an immunoblotting technique, four structural proteins were detected that could bind phage antibodies. PMID- 18425681 TI - Galactose and lactose transport in Kluyveromyces lactis. AB - Transport of lactose into Kluyveromyces lactis was accomplished by a highly specific system inducible by lactose and galactose. The biosynthesis of the transport enzyme was strongly repressed by glucose. For non-induced cells, lactose penetrated by passive transport, like galactose in any type of cells. The lactose transport showed aK (m) 1.2 -4 mm, was temperature-dependent (76 kJ/mol) and was blocked by metabolic inhibitors. PMID- 18425682 TI - Isolation and characterization of a novel methylhopanoid from a facultative methylotrophic Corynebacterium sp. AB - Several compounds such as a methylhopanoid and carotenoids have been isolated and characterized from a facultative methylotrophicCorynebacterium sp., a vitamin B(12) producer. A novel pentacyclic triterpene, 2-methyl-22-hydroxyhopane has been identified by IR,(1)H-and(13)C-NMK and mass spectrometry. During the purification procedure a red pigment has been characterized as a mixture of several carotenoids by TLC and UV-VIS spectroscopy. PMID- 18425683 TI - The influence of various carbon and nitrogen sources on oil production by Fusarium oxysporum. AB - The oil-synthesizing capacity of Fusarium oxysporum, cultivated on basal nutrient medium, was evaluated using different carbon and nitrogen sources. In one of the media, molasses was also used as a principal carbon source. Media containing glucose and ammonium nitrate were found to be most efficient for oil production. Fatty acid profile of the fungal oil indicated the presence of a wide range of fatty acids ranging from C8 to C24. Fatty acid composition largely depends on the type of carbon and nitrogen sources. PMID- 18425684 TI - Tyrosine-induced heterocyst division in Nostoc muscorum. AB - Heterocyst size variation in Nostoc muscorum has been surveyed in the presence and absence of tyrosine. The heterocyst size exhibited two major peaks under both conditions but one of the peaks shifted towards larger size in tyrosine containing medium. Heterocysts of larger volume exhibited division in the latter medium which was not observed in medium lacking tyrosine. It is suggested that signals for cell division did not decay following differentiation of heterocyst in the presence of tyrosine. PMID- 18425685 TI - Indoleacetic acid production by Rhizobium sp. from a leguminous tree, Erythrina indica. AB - Rhizobium sp. isolated from the root nodules of Erythrina indica Lamk., a leguminous tree, produced large amounts of indoleacetic acid (IAA) from tryptophan. L-Tryptophan (2 mg/ mL) was preferred to DL-tryptophan for IAA synthesis. Attempts were made to optimize the cultural requirements for an accelerated IAA production. An enrichment of the carbon-free incubation medium with maltose (1 %), MnSO4 x H2O (5 microg/mL), and Triton X-100 (100 ppm) promoted the synthesized IAA content of the medium by 480 %. PMID- 18425686 TI - Genetic recombination in auxotrophic strains of Phanerochaete chrysosporium. AB - Four auxotrophic strains of the ligninolytio basidiomycetePhanerochaete chrysosporium were obtained by UV mutagenesis. The heterokaryotic mycelium formed by complementation of different auxotrophic isolates was able to fruit and produce basidiospores. From the hasidiospore progeny of the heterokaryons prototrophic strains and strains with a recombined set of parental nutritional requirements were isolated. Genetic recombination hence takes place in fruit bodies produced by the heterokaryotic mycelium. PMID- 18425689 TI - Prevalence of Giardia lamblia with or without diarrhea in South East, South East Asia and the Far East. AB - This article is a review of the latest information on the prevalence of G. lamblia in South Asia, South East Asia and Far East, characterizing the current endemic situation within these regions. Around 33 published papers from 2002-2007 were collected on G. lamblia. The included countries were Nepal, Bangladesh, India, Cambodia, Vietnam, Malaysia, Philippines, Indonesia, Thailand, Republic of Korea, and China. Only five published papers were discarded because data was extracted before 2002-2007 or they are not included within our regions, emphasizing more on G. lamblia in animals, or performed at extensive molecular level. The prevalence of G. lamblia varied markedly between studies illustrating higher levels in the urban than in the rural areas, more among poor communities, slightly higher in males than in females with age range of 2-5-year-old children, and among university students, old-aged people, HIV-positive patients, and gastric carcinoma patients. Though G. lamblia is not a life-threatening parasite, nevertheless, it is still considered as the most common water-borne diarrhea causing disease. It is important to understand the etiology, frequency, and consequences of acute diarrhea in children. Routine surveillance such as bi annual follow-up treatments, treating G. duodenalis cysts and other protozoa oocysts detected in ground water sources, and continuous health education are the most preventive measures. PMID- 18425690 TI - Renal pathology of ANCA-related vasculitis: proposal for standardization of pathological diagnosis in Japan. AB - BACKGROUND: In Japan, systematic evaluation of the histologic parameters of anti neutrophil cytoplasmic autoantibodies (ANCA)-related vasculitis has been performed according to the Japanese classification by Shigematsu et al. However, this classification is quite different from that of the European Vasculitis Study Group (EUVAS) classification. Therefore, a histological common basis is needed to compare Japanese histological data with the international database. METHOD: Histological parameters concerning glomerular, tubulointerstitial, and vascular lesions of ANCA-related vasculitis, which are indispensable for clinical management, were elucidated and defined by reviewing, utilizing the merits of, and amending the two scoring systems. RESULTS AND CONCLUSION: A new comprehensive and standardized scoring system, by which histological quantitative assessment can provide evidence for therapy planning, has been developed for renal biopsy of Japanese ANCA-related vasculitis. PMID- 18425691 TI - Stand stratification and woody species diversity of a subtropical forest in limestone habitat in the northern part of Okinawa Island. AB - Stand stratification and woody species diversity were studied in a subtropical forest in limestone habitat in the northern part of Okinawa Island, Japan. The forest stand consisted of four architectural layers. Cinnamomum japonicum Sieb. was the most dominant species as it had the highest importance value in all layers, especially in the lower three layers. Although Rhus succedanea L. was a major top-layer species, it might disappear because of its absence in the lower two layers. The lower two layers had similar floristic composition, while the composition of the top layer differed greatly from that of the lower two layers. The value of H' in each layer increased from the bottom to the second layer and then decreased, and the value of J' in each layer consistently increased from the bottom layer upward. The values of Shannon's index H' and Pielou's index J' were 4.27 bit and 0.74, respectively, at the stand level. The trends of H' and J' based on the cumulative height range from the top layer downward were, respectively, the same as those in each layer for the present forest in limestone habitat. However, these trends were opposite from those of a nearby subtropical forest in silicate habitat, where the values of H' and J' based on the cumulative height range decreased steadily from the bottom layer upward. PMID- 18425693 TI - [Social medicine and healthcare economics. The framework for future forms of healthcare]. AB - In the political debate, even in academia, the concepts of "profitability" or "efficiency" are thrown around very robustly and freely with no regard for the players themselves. Economically speaking there can be no efficiency without a definition of targets in terms of outcomes and their level of quality. If even the government's Council of Economic Experts itself finds in its assessment of hospital funding that the "reform's target parameters - improving the profitability of service provision - have developed positively", but adds that "whether this also applies to the quality of services provided or to the realisation of healthcare outcomes remains to be seen due to the lack of evidence" [21], this indicates a one-sided and problematic curtailment of the concept even by highly competent bodies. Economic control of new forms of healthcare by means of prices and fees for clearly defined services is a complex problem that has not been dealt with adequately. All pricing is based on classification models aimed at ensuring cost and benefit clusters that are as homogeneous as possible. Classification models in healthcare as a basis for price control targets need constant adjusting to ensure accuracy of mapping and appropriateness to performance. A prerequisite for the methodology behind price control models of this kind is presupposing a responsible, rule-bound and criteria-based handling of "variance" and "coincidence" by means of risk-adjusted quality and price systems. They will define the character of a wide range of steering tools and have an effect that goes beyond the narrow formal confines of the sector. That is why the regulatory framework will need first and foremost to define a qualitative framework for the political "security infrastructure" by means of deregulated economic processes in which price control becomes accountable and is justified in terms of content. PMID- 18425694 TI - [Rare diseases. Funding programs in Germany and Europe]. AB - Rare diseases are defined by lifetime prevalence and are a medically heterogeneous group. Treatment options and the state of knowledge about these diseases are also very heterogeneous, as well as the respective needs for research. This article provides an overview on funding programs in Germany, further examples of countries within Europe and the European Commission, and a few examples of research networks. It is one of the goals of the article to show similarities and differences between the funding programs. The funding organizations of most countries and the European Commission (FP7) fund research on rare diseases with programs in which researchers define the contents in bottom up approaches. While, in general, basic science is well established, the translation of knowledge into clinical benefit for the patients is slow. Because of the low prevalence and geographic distribution of patients and researchers, research on rare diseases suffers from infrastructural deficits. While France, Germany, Italy and Spain have implemented research programs which are specific for rare diseases and aim to support networking of scarce resources and to stimulate interdisciplinary collaborations, other countries fund research on rare diseases within the context of general programs. PMID- 18425692 TI - Plant-derived natural product research aimed at new drug discovery. AB - Many important bioactive compounds have been discovered from natural sources using bioactivity-directed fractionation and isolation (BDFl) [Balunas MJ, Kinghorn AD (2005) Drug discovery from medicinal plants. Life Sci 78:431-441]. Continuing discovery has also been facilitated by the recent development of new bioassay methods. These bioactive compounds are mostly plant secondary metabolites, and many naturally occurring pure compounds have become medicines, dietary supplements, and other useful commercial products. Active lead compounds can also be further modified to enhance the biological profiles and developed as clinical trial candidates. In this review, the authors will summarize research on many different useful compounds isolated or developed from plants with emphasis placed on those recently discovered by the authors' laboratories as antitumor and anti-HIV clinical trial candidates. PMID- 18425695 TI - Hemorrhagic acute disseminated encephalomyelitis as first manifestation of systemic lupus erythematosus. PMID- 18425697 TI - Arterial wave reflection is elevated in evening hemodialysis patients. AB - AIM: Cardiovascular disease constitutes a major cause of death in patients with chronic kidney diseases and is related to enhanced arterial stiffness. It has been reported that daytime hemodialysis patients show better prognosis than evening hemodialysis patients. METHODS: Aortic augmentation index (AI) and radial AI were measured in 20 non-diabetic hemodialysis patients, using SphygmoCor (PWV Medical) and HEM-9010AI (Omron Healthcare) as markers of arterial stiffness. Cardiovascular prognosis was followed for two years. RESULTS: Mean age, blood pressure, pulse rate, and aortic and radial AI were 52 +/- 4 y/o, 142 +/- 6/74 +/ 4 mmHg, 78 +/- 4 bpm, 21 +/- 2, and 71 +/- 4%, respectively, in 10 daytime hemodialysis patients, and they averaged 52 +/- 3 y/o, 146 +/- 6/76 +/- 4 mmHg, 74 +/- 3 bpm, 26 +/- 2, and 73 +/- 3% in 10 evening hemodialysis patients, respectively. Thus, aortic AI was higher in evening hemodialysis patients (p < 0.05). In all, one patient was hospitalized due to cardiovascular complications in daytime hemodialysis patients, as well as three patients from those dialyzed in the evening. CONCLUSION: Although a larger scale study is required to draw a definite conclusion, our findings suggest that a better prognosis in daytime hemodialysis non-diabetic patients is related to the lower AI. PMID- 18425698 TI - Does short sleep duration in daily life affect morning home blood pressure? Evaluation in Japanese people. AB - A short sleep duration is expected to elevate blood pressure the next morning, but no report has evaluated this in detail using home blood pressure measurement. In this study, the relation between sleep duration and morning and evening home blood pressure and heart rate during seven consecutive days was evaluated. From 630 volunteers not receiving antihypertensive agents, we selected 478 subjects (318 male, 160 female; mean age: 39.0 years) whose 2-7 days of data consisted of 7-8 hours sleep duration (proper sleep period phase; mean sleep duration: 7.3 +/- 0.3 hours) and less than 7 hours (short sleep period phase; 5.7 +/- 4.9 hours). In the morning, systolic blood pressure and heart rate in the short sleep period phase (117.7 +/- 14.9 mmHg, 67.3 +/- 9.6/min) were significantly (p < 0.01) higher than those in the proper sleep period phase (116.9 +/- 14.9 mmHg, 66.5 +/- 9.1/min). However, there was no difference in morning diastolic blood pressure. Although the difference in morning systolic blood pressure had disappeared by the time of measurement before going to bed, the difference in heart rate was maintained (proper sleep period phase: 70.4 +/- 10.2/min, short sleep period phase: 71.7 +/- 10.7/min, p < 0.01). In conclusion, days with sleep duration of less than 7 hours showed higher morning systolic blood pressure and heart rate compared with days with sleep duration between 7 and 8 hours, but no difference was found in diastolic blood pressure. Moreover, although the difference in morning systolic blood pressure had disappeared at night, the difference in heart rate was still maintained. PMID- 18425699 TI - Increased blood pressure levels relative to subjective feelings of intensity of exercise determined with the Borg scale in male patients with hypertension. AB - We examined the hemodynamic responses to exercise and symptoms in 37 male patients with untreated essential hypertension, and compared the findings with those in 32 age-matched healthy male volunteers by performing a graded symptom limited exercise test using a bicycle ergometer. The subjective feeling of intensity of exercise was determined using the Borg scale. In the relationship between Borg scores and blood pressure (BP), patients with hypertension showed higher systolic BP and diastolic BP relative to the Borg scores than the controls. Consequently, patients with hypertension showed significantly higher systolic BP with Borg scores < or = 3 (subjective symptoms < or = moderately hard) than the controls (177.8 +/- 27.0 vs. 143.7 +/- 17.9 mmHg, p < 0.0001). Similarly, significantly higher diastolic BP with Borg scores < or = 3 was observed in patients with hypertension than in the controls (101.6 +/- 12.0 vs. 82.6 +/- 11.6 mmHg, p < 0.0001). The pulse pressure with Borg scores < or = 3 was also significantly higher in patients with hypertension than in the controls (76.2 +/- 20.6 vs. 61.0 +/- 13.6 mmHg, p < 0.0001). Hypertensive patients showed a decrease in the high-frequency power of heart rate variability at initial low load exercise. In conclusion, the present study revealed that there was a greater BP response relative to the Borg score in patients with hypertension than in the controls. Autonomic nerve activity may contribute to some extent to these different relations. A determination of the relationship between the subjective feeling of intensity of the exercise and BP levels caused by a given intensity of load is essential before exercise training in patients, at least in males, with hypertension to avoid increasing the risk of cardiovascular events in association with excessive exercise training. PMID- 18425700 TI - Masked hypertension in elderly managerial employees and retirees. AB - Masked hypertension is reported to have the same level of hazard risk of cardiovascular mortality and stroke morbidity as sustained hypertension. The number of managerial employees suffering from cardiovascular disease and stroke is known to be greater than other employee. The aim of this study was to compare the 24-h blood pressure (BP) recordings between elderly male managerial employees and retirees and to propose a strategy for identifying masked hypertension. A total of 38 males (16 managerial employees aged 50-69 years and 22 retirees aged 60-65 years) who were not taking any antihypertensive medications participated in this study. Their 24-h BP was measured by an ambulatory BP monitoring device. Daytime (9:00-17:00 h) BPs of the employees (mean, 139/92 mm Hg) were significantly higher than in the retirees (mean 124/80 mm Hg), while there was no difference in BP before and during sleep. In all, 5 of 16 employees (31%) who were diagnosed as normotensive (<140/90 mm Hg) at a periodic health check had hypertension (>135/85 mm Hg) in the morning measured by ambulatory BP monitoring, while 6 (38%) had a similar level of hypertension during the daytime (9:00-17:00 h). These individuals were diagnosed as having masked hypertension. Multiple regression analyses showed that the job was the only factor that contributed to the difference in BP in the subjects during the daytime. This finding suggested that job stress seemed to be one of the main causes of masked hypertension. We argue that more frequent measurements of BP at the work place are necessary to identify subjects with masked hypertension. PMID- 18425701 TI - Ambulatory blood pressure variability is increased in diabetic hypertensives. AB - The purpose of this study was to examine the possible difference in the 24-hr BP profile--including short-term BP variability, assessed as the standard deviation- between diabetic and non-diabetic hypertensives. We measured 24-hr ambulatory BP in 11 diabetic hypertensives (diabetic HT) and 10 non-diabetic hypertensives (non diabetic HT) who were hospitalized for the educational program in our hospital and were under stable salt intake. Renal function and sleep apnea were also estimated. There were no significant differences in 24-hr systolic BP (141 mmHg vs. 135 mmHg, ns), daytime systolic BP (143 mmHg vs. 138 mmHg, ns), and nighttime systolic BP (135 mmHg vs. 130 mmHg, ns) between diabetic HT and non-diabetic HT. The values of 24-hr HR (69.7 beats/min vs. 65.2 beats/min, ns) and 24-hr HR variability (9.9 beats/min vs. 10.1 beats/min, ns) were also similar between the groups. Interestingly, diabetic HT had a significantly greater 24-hr systolic and diastolic BP variability than non-diabetic HT (18.2 mmHg vs. 14.5 mmHg, p < 0.05; 11.5 mmHg vs. 9.6 mmHg, p < 0.05, respectively). The values for creatinine clearance, urinary protein excretion, and apnea-hypopnea index were similar between the groups. Bivariate linear regression analysis demonstrated that fasting blood glucose was the primary determinant of 24-hr diastolic BP variability (r = 0.661, p < 0.01). Multiple stepwise regression analysis revealed that fasting blood glucose was a significant and independent contributor to 24-hr systolic BP variability (r = 0.501, p < 0.05). Taken together, these results demonstrate that BP variability is increased in diabetic hypertensives. Furthermore, it is possible that an elevation of fasting blood glucose may contribute to the enhanced BP variability in hypertensives. PMID- 18425702 TI - Home blood pressure measurement may lead to less strict control of office blood pressure. AB - Home blood pressure (HBP) measurement is useful for detecting morning hypertension, white coat as well as masked hypertension. However, target BP levels based on HBP remain unknown. The purpose of the present study was to evaluate the relationship between HBP measurement and office BP control status in hypertensive patients. Subjects were a total of 720 hypertensive outpatients (mean age: 64 +/- 11 years; females: 57%). Two-time averaged office BP in 2005 were categorized as excellent (<130/85 mmHg), good (> or =130/85 and <140/90 mmHg), or poor (>140/90 mmHg) control. In all patients, 37% were classified as excellent, 37% as good, and 26% as poor control. A total of 393 (55%) patients regularly measured HBP (HBP group). More women belonged to the HBP group (62 vs. 52%, p < 0.05). The HBP group also showed lower body mass index (23.8 +/- 3.3 vs. 24.7 +/- 3.4 kg/m(2), p < 0.01), lower triglyceride (136 +/- 78 vs. 158 +/- 89 mg/dl, p < 0.01), and lower blood glucose (104 +/- 20 vs. 118 +/- 42 mg/dl, p < 0.01). HBP group showed a significantly higher prevalence of poor BP control (33 vs. 23%, p <0.01) and higher office SBP (134.5 +/- 14.5 vs. 131.3 +/- 11.7 mmHg, p < 0.01) than those who did not measure HBP (non-HBP). In a multivariate analysis for office SBP, age (partial r = 0.21, p < 0.05) and HBP measurement (partial r = 0.12, p < 0.05) were detected as significant independent variables. These results suggest that HBP measurement may lead to less strict office BP control unless the target HBP levels are clearly indicated. Until the recommendations or target HBP levels are available, we should make an effort to obtain goal office BP. PMID- 18425703 TI - Influence of mental stress on the plasma homocysteine level and blood pressure change in young men. AB - Objective. This study aimed to determine whether mental stress influences the plasma total homocysteine level or blood pressure in young men. Method. Twenty seven male university students were assigned to a normal blood pressure group (24 h systolic blood pressure <125 mmHg and diastolic blood pressure <75 mmHg; 13 subjects) or a high blood pressure group (24-h systolic blood pressure > or =125 mmHg, or 24-h diastolic blood pressure > or =75 mmHg; 14 subjects). Wearing an ambulatory blood pressure monitoring device, subjects rested for 30 minutes, underwent an arithmetic test for 15 minutes, and rested again for 15 minutes. Blood samples were taken before and after the test. Plasma total homocysteine levels were measured. Heart rate, blood pressure, and sympathovagal balance were determined during the test. Results. The mean total homocysteine level at rest in the high blood pressure group was slightly, but not significantly, higher than that in the normal blood pressure group. The resting total homocysteine level was significantly higher in subjects with parental history of hypertension than in those without (p < 0.01). Blood pressure, heart rate, and the plasma total homocysteine level were increased significantly by mental stress (p < 0.05). The change in total homocysteine correlated significantly with the changes in systolic blood pressure and sympathovagal balance (p < 0.05). Conclusion. Resting total homocysteine level was significantly higher in male students with a parental history of hypertension than in those without. It was shown that mental stress elevates heart rate, blood pressure, sympathovagal activity, and the plasma total homocysteine level in young men. PMID- 18425704 TI - Time-dependent effects of imidapril administration in patients with morning hypertension measured as home blood pressure. AB - We investigated the effect of the timing of imidapril administration in patients with morning hypertension (home blood pressure [HBP] in the morning [morning HBP] > or =135/85 mmHg). Eighty-seven patients (mean age, 61 years; 48% women) with morning hypertension each measured HBP in the early morning, before going to bed for the final five days of the observation, and during treatment. Imidapril (2.5 or 5.0 mg) was administered once every morning (n = 57) or at bedtime (n = 30) for four weeks (monotherapy: n = 30, 34%; combined with other antihypertensive drugs: n = 57, 66%). The morning (M) versus evening (E) effect (M/E ratio) and the evening versus morning effect (E/M ratio) were calculated to assess the duration of imidapril action. Morning HBP was significantly reduced (all p < 0.05) except when 2.5 mg of imidapril was administered once at bedtime. The individual M/E ratios expressed as means +/- standard deviation for systolic HBP were 0.9 +/- 0.7 and 0.7 +/- 0.7 for the morning administration of 2.5 and 5.0 mg of imidapril, respectively. The corresponding E/M ratios for bedtime administration were 0.3 +/- 0.5 and 1.0 +/- 0.7, respectively. One daily dose of imidapril taken either in the morning or at bedtime lowered morning HBP, indicating that imidapril is useful for controlling morning hypertension. PMID- 18425705 TI - Determinants of self-measured pulse rate profile in medicated hypertensives: the Jichi Morning Surge-1 (JMS-1) study. AB - BACKGROUND: Recently, it was reported that high pulse rate (PR), which was measured using by self-measured blood pressure (BP) monitoring at home, was associated with cardiovascular risk. However, the predictor of high PR at home in treated hypertensives is unknown. In this study, we studied the predictor of high PR and evaluated pulse rate variability (PRV). METHODS: In the JMS-1 study, 611 hypertensive outpatients were recruited. Self-measured BP monitoring was conducted consecutively twice in the morning and evening for three days. PR analysis was conducted using the average of these two measurements for three days (six readings in total). We defined home PR as the mean of these six readings. Home PRV was defined as the standard deviation of these six readings. RESULTS: Multivariate linear regression analysis demonstrated that current smoking (beta = 0.12, p = 0.002), diabetes (beta = 0.16, p < 0.001), lack of angiotensin converting enzyme (ACE) inhibitor use (beta = 0.10, p = 0.008), decreased brain type natriuretic peptide (BNP; beta = 0.17, p < 0.001), and elevated home diastolic blood pressure (beta = 0.14, p = 0.009) were determinants of high PR. Determinants of decreased home PRV were female gender (beta = 0.10, p < 0.03) and increased hemoglobin A1c (HbA1c; beta = 0.15, p < 0.001). When we divided the patients into four groups according to home PR and its variability, hypertensives whose home PR was high and variability was low were found to have high HbA1c (ANOVA, p > 0.05). CONCLUSIONS: Smoking habit, diabetes, lack of ACE inhibitor use, and low BNP value were determinants of home PR, and female gender and higher HbA1c were significantly associated with its low variability. Home PR and its variability may be useful for detecting high-risk hypertensive patients, particularly with autonomic neuropathy. PMID- 18425706 TI - Effects of valsartan or amlodipine on endothelial function and oxidative stress after one year follow-up in patients with essential hypertension. AB - Endothelial function is impaired in hypertensive patients. Decreased nitric oxide production and increased oxidative stress are involved in this abnormality. The aim of the present study was to evaluate whether endothelial function and oxidative stress differ following long-term antihypertensive treatment with an angiotensin type 1 receptor blocker, valsartan, or a calcium channel blocker, amlodipine, in patients with essential hypertension. Hypertensive patients were treated with valsartan (80-160 mg/day) or amlodipine (5-10 mg/day) for one year (n = 9 for each). The baseline blood pressure was similar between groups, and the magnitude of the decreases in blood pressure did not differ during treatment at three months, six months, or one year. Endothelial function and oxidative stress markers were examined before and after treatment. Endothelial function, assessed by flow-mediated vasodilation, was significantly improved in hypertensive patients treated with valsartan (5.8 +/- 1.2 to 10.7 +/- 1.4 %, p < 0.01) but not in those treated with amlodipine. The percent increase in vasodilation induced by sublingual nitroglycerin did not differ between the two groups. As markers of oxidative stress, urinary excretion of 8-isoprostane and 8-hydroxy-2' deoxyguanosine was significantly reduced in patients treated with valsartan, but not in those treated with amlodipine. These findings suggest that the treatment of hypertensive patients with valsartan for at least one year improves endothelial function in association with reduced oxidative stress. The improved endothelial function and reduced oxidative stress might be involved in the benefits of anti-hypertensive treatment beyond simply lowering blood pressure, although the effects of treatment with valsartan or amlodipine over a much longer period are unknown. PMID- 18425707 TI - Prevalence of Masked Hypertension in Subjects Treated with Antihypertensive Drugs as Assessed by Morning versus Evening Home Blood Pressure Measurements: the J HOME study. AB - To investigate the relationship between morning and evening home blood pressure (BP) measurements to make a diagnosis of masked hypertension, we collected information on the characteristics of 3,303 essential hypertensive outpatients receiving antihypertensive medication in Japan using a physician, self administered questionnaire. All patients were asked to measure their home BP once every morning and once every evening for two weeks. Morning and evening home BP values of each patient were defined as the average of all morning and all evening home BP values, respectively. The mean BP values of all study subjects were 142.8/80.6 mmHg for office BP, 139.8/81.8 mmHg for morning home BP, 133.7/76.9 mmHg for evening home BP, and 136.8/79.3 mmHg for the average of the morning and evening home BPs. Masked hypertension was defined as an office BP < 140/90 mmHg and a home BP > or = 135/85 mmHg. The prevalence of masked hypertension diagnosed using morning home BP (23.1%) was higher than that diagnosed by evening home BP (14.7%); the prevalence was 19.0% when diagnosed using the average of the morning and evening home BPs. Among the 1,386 patients with a normal office BP, the diagnosis of masked hypertension based on morning and evening home BP values differed in 28.8% of patients for systolic BP and 20.9% for diastolic BP (kappa coefficient = 0.43). The present study showed that the prevalence of masked hypertension was underestimated when the diagnosis of masked hypertension was made on the basis of evening home BP. PMID- 18425708 TI - Masked hypertension: subtypes and target organ damage. AB - Masked hypertension has been drawing attention recently because this condition is often seen in untreated and treated individuals and is associated with target organ damage and a poor cardiovascular prognosis. Although masked hypertension is defined as normal office blood pressure with elevated ambulatory or home blood pressure, there are several subtypes. Morning hypertension is the most common form of masked hypertension, and is caused by natural circadian variation, evening alcohol consumption, and the use of short-acting antihypertensive drugs. Daytime hypertension may be caused by lifestyle factors such as habitual smoking and mental or physical stress. Nighttime hypertension is seen in various conditions that produce non-dipping status, including a high salt intake, renal dysfunction, obesity, sleep apnea, and autonomic failure. Advanced target organ damage such as increases in the left ventricular mass, carotid artery intima media thickness, and urinary albumin excretion, is often present both in untreated and treated subjects with masked hypertension. In our study, the presence of the reverse white-coat effect is independently associated with those indices of organ damage among treated hypertensive patients. It is important to identify individuals with masked hypertension, to evaluate them with including the search for the subtype, and to treat each patient appropriately according to the cause of this condition. PMID- 18425709 TI - Patterns of natural herb use by Asian and Pacific Islanders. AB - OBJECTIVE: Previous studies have noted the role of race/ethnicity in use of complementary and alternative medicine (CAM). Asian and Pacific Islanders (APIs) in particular have been found to use herbs more frequently, although the patterns of use among this population have not been described. The goal of this study was to characterize the rates and patterns of herb use among the API population of the United States. DESIGN: Data from the 2002 US National Health Interview Survey, including the supplement on CAM, were analyzed using SAS. Cross tabulation and logistical regression were used to determine the association between herb use and race/ethnicity, while adjusting for demographic factors. RESULTS: After adjusting for age, gender and educational attainment, we found that APIs were more likely to use herbs than non-Hispanic Whites (OR=1.3; 95% CI: 1.1, 1.5; p=0.013), with the difference being most prominent in those 60 years old or older (OR=2.9; 95% CI: 1.8, 4.5; p<0.0001). Herb use among APIs was more common among women, those with higher education and income, and those in older age groups. Patterns of use also varied among API subgroups, with Chinese Americans reporting the highest rates of herb use in their lifetimes (44%) and in the previous 12 months (33%). CONCLUSION: Considerable diversity exists within API subgroups with regard to the prevalence and demographic and health correlates of natural herb use. Herb use is more common among APIs than non-Hispanic Whites, particularly among the elderly. The disaggregation of APIs by racial/ethnic subgroups may be important for patient-health care professional interactions, and when designing studies of racial/ethnic minority populations. PMID- 18425710 TI - 'Race' matters: racialization and egalitarian discourses involving Aboriginal people in the Canadian health care context. AB - The major purpose of this paper is to examine how 'race' and racialization operate in health care. To do so, we draw upon data from an ethnographic study that examines the complex issues surrounding health care access for Aboriginal people in an urban center in Canada. In our analysis, we strategically locate our critical examination of racialization in the 'tension of difference' between two emerging themes, namely the health care rhetoric of 'treating everyone the same,' and the perception among many Aboriginal patients that they were 'being treated differently' by health care providers because of their identity as Aboriginal people, and because of their low socio-economic status. Contrary to the prevailing discourse of egalitarianism that paints health care and other major institutions as discrimination-free, we argue that 'race' matters in health care as it intersects with other social categories including class, substance use, and history to organize inequitable access to health and health care for marginalized populations. Specifically, we illustrate how the ideological process of racialization can shape the ways that health care providers 'read' and interact with Aboriginal patients, and how some Aboriginal patients avoid seeking health care based on their expectation of being treated differently. We conclude by urging those of us in positions of influence in health care, including doctors and nurses, to critically reflect upon our own positionality and how we might be complicit in perpetuating social inequities by avoiding a critical discussion of racialization. PMID- 18425711 TI - Ethnic differences in self-rated and functional health: does immigrant status matter? AB - The current study examines self-rated health status and functional health differences between first-generation immigrant and Canadian-born (CB) persons who share the same ethnocultural origin, and the extent to which such differences reflect social structural and health-related behavioural contexts. Multivariate analyses of data from the 2000/2001 Canadian Community Health Survey indicate that first-generation immigrants of Black and French ethnicity tend to have better health than their CB counterparts, while the opposite is true for those of South Asian and Chinese origins, providing evidence that for these groups, immigrant status matters. West Asians and Arabs and other Asian groups are advantaged in health regardless of country of birth. Health differences between ethnic foreign-born and CB persons generally converge after controlling for sociodemographic, socioeconomic status (SES), and lifestyle factors. Analysis of the data does however reveal extensive ethnocultural disparities in self-rated and functional health within both the immigrant and CB populations. Implications for health care policy and programme development are discussed. PMID- 18425712 TI - Immigrant advantage? Substance use among Latin American immigrant and native-born youth in Spain. AB - This article reports the results of a descriptive study conducted with middle school and high school age youth residing in northwestern Spain. The main outcome of the study is to advance knowledge about the drug use attitudes and behaviors of immigrants versus native youth in a social context where Latin American immigrants share a common language and a set of core cultural norms with the host society. The research was conducted by a bi-national Spain-US research team as a preliminary study leading to the development of joint culturally appropriate prevention interventions for youth in the northern region of Galicia, Spain. Surveys were administered in Spring 2005 to 817 students in 7th to 10th grades in 10 urban, secondary schools with high immigrant enrollment. The sample included Spanish natives (two-thirds) and Latin American immigrants (one-third), mainly from Colombia, Argentina, and Venezuela. Multiple regression analyses predicted substance use intentions, and a composite variable measuring lifetime and last 30 day frequency and amount of alcohol, cigarette and marijuana use. Controlling for the fact that the immigrant students were generally older and performing less well academically than natives, and for other predictors, Latin American immigrant youth were less at risk than native youth on their intentions to use substances and on their reported actual substance use. In a mediational analysis, most of the key explanatory variables in youth substance use etiology failed to account for the immigrant versus native differences, including a range of risk and protective factors for substance use, substance use norms, strength of ethnic identity, and degree of social integration within native-born social networks. Differential access to drugs mediated the immigrant-native gap in substance use intentions but did not mediate differences in actual substance use. PMID- 18425713 TI - Small-group discussions on menthol cigarettes: listening to adult African American smokers in Atlanta, Georgia. AB - OBJECTIVE: In 2002, the First Conference on Menthol Cigarettes brought together researchers from diverse backgrounds to summarize what is known about menthol cigarettes and the people who smoke them, and to identify areas of needed research on menthol cigarettes. Since the conference, PubMed reports 24 articles, including the conference proceedings, on menthol cigarettes and African Americans. Many of the articles address epidemiological or biomedical topics. While there has been some focus on social influences and marketing issues, more research and a greater focus on this topic are needed. DESIGN: To stimulate research on a population disproportionately burdened by the health effects of smoking, we conducted small-group discussions in 2005 with adult African American smokers in Atlanta, Georgia. Each group discussion focused on a different topic: smoking behavior and preferences, perceptions of social influences, health effects and perceived harmfulness of menthol, quitting menthol cigarette smoking, or the influence of marketing and advertising of menthol cigarettes. RESULTS: Themes emerged from the discussions: (1) emulation of black culture by white youth and racial integration of neighborhoods and communities may have modified the perception that African Americans smoke menthol cigarettes and whites smoke non-menthol cigarettes; (2) non-menthol cigarette smokers were thought to be 'hardcore' smokers with less interest in quitting; (3) switching to non-menthol cigarettes was discussed as a way of quitting cigarettes for habitual menthol smokers; and, (4) smoking menthol cigarettes was thought to lead to fewer negative health effects. CONCLUSION: Some topics suggested by the participants warrant further investigation. More research is needed to assess the pervasiveness of these beliefs and their potential utility for smoking cessation interventions. PMID- 18425714 TI - Phylogenetic analysis informed by geological history supports multiple, sequential invasions of the Mediterranean Basin by the angiosperm family Araceae. AB - Despite the remarkable species richness of the Mediterranean flora and its well known geological history, few studies have investigated its temporal and spatial origins. Most importantly, the relative contribution of geological processes and long-distance dispersal to the composition of contemporary Mediterranean biotas remains largely unknown. We used phylogenetic analyses of sequences from six chloroplast DNA markers, Bayesian dating methods, and ancestral area reconstructions, in combination with paleogeographic, paleoclimatic, and ecological evidence, to elucidate the time frame and biogeographic events associated with the diversification of Araceae in the Mediterranean Basin. We focused on the origin of four species, Ambrosina bassii, Biarum dispar, Helicodiceros muscivorus, Arum pictum, subendemic or endemic to Corsica, Sardinia, and the Balearic Archipelago. The results support two main invasions of the Mediterranean Basin by the Araceae, one from an area connecting North America and Eurasia in the Late Cretaceous and one from the Anatolian microplate in western Asia during the Late Eocene, thus confirming the proposed heterogeneous origins of the Mediterranean flora. The subendemic Ambrosina bassii and Biarum dispar likely diverged sympatrically from their widespread Mediterranean sister clades in the Early-Middle Eocene and Early-Middle Miocene, respectively. Combined evidence corroborates a relictual origin for the endemic Helicodiceros muscivorus and Arum pictum, the former apparently representing the first documented case of vicariance driven by the initial splitting of the Hercynian belt in the Early Oligocene. A recurrent theme emerging from our analyses is that land connections and interruptions, caused by repeated cycles of marine transgressions-regressions between the Tethys and Paratethys, favored geodispersalist expansion of biotic ranges from western Asia into the western Mediterranean Basin and subsequent allopatric speciation at different points in time from the Late Eocene to the Late Oligocene. PMID- 18425715 TI - Parsimony via consensus. AB - The parsimony score of a character on a tree equals the number of state changes required to fit that character onto the tree. We show that for unordered, reversible characters this score equals the number of tree rearrangements required to fit the tree onto the character. We discuss implications of this connection for the debate over the use of consensus trees or total evidence and show how it provides a link between incongruence of characters and recombination. PMID- 18425716 TI - Accounting for phylogenetic uncertainty in biogeography: a Bayesian approach to dispersal-vicariance analysis of the thrushes (Aves: Turdus). AB - The phylogeny of the thrushes (Aves: Turdus) has been difficult to reconstruct due to short internal branches and lack of node support for certain parts of the tree. Reconstructing the biogeographic history of this group is further complicated by the fact that current implementations of biogeographic methods, such as dispersal-vicariance analysis (DIVA; Ronquist, 1997), require a fully resolved tree. Here, we apply a Bayesian approach to dispersal-vicariance analysis that accounts for phylogenetic uncertainty and allows a more accurate analysis of the biogeographic history of lineages. Specifically, ancestral area reconstructions can be presented as marginal distributions, thus displaying the underlying topological uncertainty. Moreover, if there are multiple optimal solutions for a single node on a certain tree, integrating over the posterior distribution of trees often reveals a preference for a narrower set of solutions. We find that despite the uncertainty in tree topology, ancestral area reconstructions indicate that the Turdus clade originated in the eastern Palearctic during the Late Miocene. This was followed by an early dispersal to Africa from where a worldwide radiation took place. The uncertainty in tree topology and short branch lengths seems to indicate that this radiation took place within a limited time span during the Late Pliocene. The results support the role of Africa as a probable source area for intercontinental dispersals as suggested for other passerine groups, including basal diversification within the songbird tree. PMID- 18425717 TI - Ocimum sanctum: a therapeutic role in diabetes and the metabolic syndrome. PMID- 18425718 TI - Altered levels of adiponectin and adiponectin receptors may underlie the effect of ciliary neurotrophic factor (CNTF) to enhance insulin sensitivity in diet induced obese mice. PMID- 18425719 TI - P-Glycoprotein inhibitory activity of lipophilic constituents of Echinacea pallida roots in a human proximal tubular cell line. AB - The N-hexane root extracts from Echinacea pallida, Echinacea angustifolia and Echinacea purpurea were evaluated for inhibition of the multidrug transporter P glycoprotein (Pgp) activity, the product of the ABCB1 gene, involved in cancer multidrug resistance (MDR) and in herb-drug or drug-drug interactions. The biological assay was performed using the human proximal tubule HK-2 cell line that constitutively expresses ABCB1. The N-hexane extracts of all three species reduced the efflux of the Pgp probe calcein-AM from HK-2 cells two-fold in a concentration-dependent manner, and E. pallida was found to be the most active species. For the first time, two polyacetylenes and three polyenes, isolated from the N-hexane extract of E. pallida roots by a bioassay-guided fractionation, were found to be able to reduce Pgp activity. Pentadeca-(8 Z,13 Z)-dien-11-yn-2-one was the most efficient compound, being able to decrease the calcein-AM efflux about three-fold with respect to the control at 30 microg/mL. PMID- 18425720 TI - Multidimensional system biology: genetic markers and proteomic biomarkers of adverse pregnancy outcome in preterm birth. AB - Premature birth before 37 weeks of gestation is a significant public health problem. Each year, 4.5 million premature infants are born worldwide. Despite extensive research and a variety of interventions, the rate of preterm birth has steadily increased over the past 20 years and reached a high of 12.8% in 2006. The etiology of most preterm births remains elusive and is likely multifactorial, with many pathophysiological pathways involved, such as excessive stretching, oxidative stress, decidual hemorrhage, and infection. Genomics and proteomics have emerged to provide a better comprehension of the pathophysiological conditions leading to preterm birth, thereby providing a perspective for improving neonatal outcome. PMID- 18425721 TI - Plasma exchange in a case of severe factor X deficiency in pregnancy: critical review of the literature. AB - Hereditary factor X deficiency represents an uncommon challenge in pregnancy. A 30-year-old primigravida affected by severe factor X deficiency was followed from 6 weeks of gestation until delivery. Factor X was provided prior to delivery for the first time in pregnancy via plasma exchange. The pregnancy and postpartum period were not complicated by bleeding episodes; therefore this approach was accompanied by lower cost and fewer side effects when compared with fresh-frozen plasma and prothrombin complex concentrates infusion, two therapeutic options already used in pregnancy. PMID- 18425722 TI - Salbutamol versus cation-exchange resin (kayexalate) for the treatment of nonoliguric hyperkalemia in preterm infants. AB - Our objective was to compare the efficacy and safety of rectal cation-exchange resin (Kayexalate) versus salbutamol infusion for the treatment of nonoliguric hyperkalemia (NOHK) in preterm infants. Data of all neonates born with NOHK during the study period of 6 years and 8 months were recorded. Diagnostic criteria of NOHK included serum potassium (SK) concentration > or = 7 mmol/L during the first 72 hours of life with urine output > or = 1 mL/kg/hour. This before-after study was divided according to the date of admission; the first 15 patients were treated with Kayexalate enema 1 g/kg every 4 hours, and the remaining 30 patients were treated with intravenous salbutamol infusion as 4 mug/kg every 4 hours. Treatment discontinued when SK became < 6 mmol/L. SK was measured every 4 hours. Daily urine was collected. Fluid intake and output, serum electrolytes, urea, creatinine, and glucose concentrations were obtained in all infants every 12 hours. All infants were observed with a cardiorespiratory monitor and oxygen saturation and blood pressure measurements. Perinatal characteristics in both groups were comparable. Mean gestational age was 26 and 28 weeks for salbutamol and Kayexalate, respectively. The peak of SK ranged between 7 and 9.3 mmol/L in the Kayexalate group and between 7 and 8.7 mmol/L in the salbutamol group ( P = 0.64). At 12 hours of treatment, SK became normal in only 4 patients (26%) in the Kayexalate group compared with 18 patients (60%) in the salbutamol group ( P = 0.003). The number of doses of Kayexalate administration was significantly higher than the doses of salbutamol ( P = 0.003). No significant side effects were detected in the salbutamol-treated infants. In contrast, there were two cases of severe ventricular tachycardia and one case of intestinal obstruction in the cation-exchange resin group. We concluded that salbutamol infusion is more effective with faster action and safer than cation-exchange resin (Kayexalate) for the treatment of NOHK in preterm infants. PMID- 18425724 TI - Integration of immunological aspects in the European Human Embryonic Stem Cell Registry. AB - The immunological properties of stem cells are of increasing importance in regenerative medicine. Immunomodulatory mechanisms seem to play an important role not only with respect to the understanding of underlying mechanisms of autologous versus allogenic therapeutic approaches, but also for endogeneous tissue regeneration. The newly established European human embryonic stem cell registry (hESCreg) offers an international database for the registration, documentation and characterisation of human embryonic stem cells (hESC) and their use. By doing so, hESCreg aims to develop a model procedure for further standardisation efforts in the field of stem cell research and regenerative medicine, and eventually the registry may lead to a repository of therapy-related information. Currently the stem cell characterisation data acquired by the registry are divided into several categories such as cell derivation, culture conditions, genetic constitution, stem cell marker expression and degree of modification. This article describes immunological aspects of stem cell characterisation and explores the layout and relevance of a possible additional section to the hESCreg repository to include immunological characteristics of human embryonic stem cells. PMID- 18425725 TI - Regulated release of B cell-derived exosomes: do differences in exosome release provide insight into different APC function for B cells and DC? AB - The induction of CD4 T(H) cell activity is a crucial component of the mammalian acquired immune response. In order to activate T(H) cells, pathogen-derived peptides are displayed on the plasma membrane of specialized cells termed APC. As well as unravelling common mechanisms in this process, considerable attention has been given to the distinct roles of the various cell types involved. In this issue of the European Journal of Immunology, a study examining the release by B cells of small vesicles termed exosomes is presented. This commentary reports that the control of exosome release from B cells appears to differ from that seen in DC. How these differences may relate to important features that differentiate the antigen presenting cell function of B cells and DC in vivo is also discussed. PMID- 18425726 TI - Activation of myelin reactive T cells in multiple sclerosis: a possible role for T cell degeneracy? AB - While it is widely accepted that myelin reactive T cells are key players in the immunopathogenesis of multiple sclerosis (MS), the initial triggers that turn on these self-reactive T-cells remain to be determined. One mechanism, which is already text book knowledge for a decade, is molecular mimicry by which viral or microbial antigens are able to cross-activate T cells specific for myelin epitopes, the major target in MS pathology. Although proof of concept for this principle was given in animal model studies, evidence for such a mechanism in MS is limited. In this issue, Zhang et al. demonstrate an increased frequency of T cells that cross-react with a wide variety of antigens, a phenomenon termed as T cell degeneracy. While the role of these degenerate T cells in MS remains to be determined the authors now provide new elegant tools to study this T cell population, thus providing a starting point to better understand their function in MS. PMID- 18425728 TI - Comparative proteomic analysis of mouse livers from embryo to adult reveals an association with progression of hepatocellular carcinoma. AB - To identify potential oncofetal biomarkers that distinguish hepatocellular carcinoma (HCC) from healthy liver tissues, we compared and analyzed the proteomic profiles of mouse livers at different developmental stages. Fetal (E13.5, E16.5), newborn (NB), postnatal (3-week) and adult (3-month) livers were isolated and profiled by 2-D PAGE. Statistical analysis using linear regression and false discovery rate (FDR) revealed that 361 protein spots showed significant changes. Unsupervised hierarchical tree analysis segregated the proteins into fetal, NB, and postnatal-adult clusters. Distinctive protein markers were identified by MALDI-TOF/MS and the corresponding mRNA profiles were further determined by Q-PCR. Fetal markers (hPCNA, hHSP7C, hHEM6) and postnatal-adult markers (hARGI1, hASSY, hBHMT, hFABPL) were selected for testing against a panel of seven human hepatocyte/HCC cell lines and 59 clinical specimens. The fetal proteins were found to be overexpressed in the metastatic HCC cell lines and the tumor tissues, whereas the postnatal-adult proteins were expressed in non-tumor tissues and normal hepatocytes. This "Ying-Yang" pattern, as orchestrated by distinct fetal and adult markers, is hypothesized to indicate the progressive change of the liver from a growing, less-differentiated organ into a functional metabolic center. Thus, embryogenesis and tumorigenesis share certain oncofetal markers and adult "hepatic" phenotypes are lost in HCC. PMID- 18425727 TI - Dendritic cells derived from TBP-2-deficient mice are defective in inducing T cell responses. AB - Thioredoxin-binding protein-2 (TBP-2), also known as vitamin D3-up-regulated protein 1 (VDUP1), was identified as an endogenous molecule interacting with thioredoxin (TRX). Here, we show that dendritic cells (DC) derived from TBP-2 deficient mice are defective in the function of T cell activation. To compare TBP 2(-/-) DC function with wild-type (WT) DC, we stimulated DC with lipopolysaccharide (LPS). Although TBP-2(-/-) DC and WT DC expressed comparable levels of MHC class II and costimulatory molecules such as CD40, CD80 and CD86, the IL-12p40, IL-12p70 and IL-6 productions of TBP-2(-/-) DC were attenuated. In a mixed leukocyte reaction (MLR), the concentrations of IL-2, IFN-gamma, IL-4 and IL-10 in the culture supernatant of MLR with TBP-2(-/-) DC were significantly lower than those in the cultures with WT DC. In MLR also, as with LPS stimulation, IL-12p40 and IL-12p70 production from TBP-2(-/-) DC was less than that from WT DC. Proliferation of T cells cultured with TBP-2(-/-) DC was poorer than that with WT DC. In vivo delayed-type hypersensitivity responses in TBP-2(-/ ) mice immunized with ovalbumin were significantly reduced compared to WT mice. These results indicate that TBP-2 plays a crucial role in DC to induce T cell responses. PMID- 18425729 TI - Malformation of true bug (Heteroptera): a phenotype field study on the possible influence of artificial low-level radioactivity. AB - The results of extensive field studies on the malformation of Western European true bugs (Heteroptera) are reviewed. More than 16,000 individuals were collected over two decades, and subjected to detailed visual inspection. Various types of disturbances were found and illustrated in detail. Depending on country, region, as well as local influences, severe disturbances and high degrees of malformation were noticed, especially in the sphere of nuclear-power installations in Switzerland (Aargau), France (La Hague), and Germany (Gundremmingen). Malformation reached values as high as 22 and 30% for morphological (MD) and total disturbance (TD), respectively. This is far above the values expected for natural populations (ca. 1%) or those determined for true bugs living in biotopes considered as relatively 'intact' (1-3%). A detailed chi-square test of the malformation data obtained for 650 true bugs from 13 collection sites near the nuclear-reprocessing plant La Hague showed a highly significant correlation (p=0.003) between malformation and wind exposure/local topography. Similar observations were made for other study sites. Currently, our data are best rationalized by assuming a direct influence between the release of anthropogenic radionuclides such as tritium ((3)H), carbon-14 ((14)C), or iodine-131 ((131)I), constantly emitted by nuclear-power and nuclear-reprocessing plants, as well as by Chernobyl and bomb-testing fallout, which is rich in caesium-137 ((137)Cs) and other long-lived noxious isotopes that have entered the food chain. The present work supports the growing evidence that low-level radiation, especially in the form of randomly scattered 'hot' alpha- and beta-particles, mainly transported via aerosols, puts a heavy burden on the biosphere in general, and on true bugs in particular. These insects could, thus, serve as sensitive 'bio-indicators' for future studies. PMID- 18425730 TI - B cell activation regulates exosomal HLA production. AB - Exosomes are nanovesicles produced constitutively and inducibly by several types of cells. They are generated as intraluminal vesicles of multivesicular bodies and express MHC and several endosomal/lysosomal proteins. In spite of their potential role in cellular immunity, the regulatory mechanisms of exosome production are largely unknown. In this study, we have established a novel ELISA system to quantify exosomal HLA using a combination of anti-HLA class I and anti HLA-DR mAb. We found that exosomal HLA production of B cells was enhanced by contact with CD4(+) T cells. Neutralizing anti-CD154 (CD40L) mAb inhibited this effect, and a soluble CD40L significantly increased production of exosomal HLA in B cells. In addition, B cell stimulation via BCR and TLR9 enhanced their production while IL-4 stimulation alone failed to do so. Strikingly, an inhibitor of the classical NF-kappaB pathway drastically inhibited exosomal HLA production in stimulated B cells, indicating that the classical NF-kappaB pathway is critical for exosomal HLA production in B cells. Together, these findings suggest a pivotal role of B cell activation in exosomal HLA production in vivo. PMID- 18425732 TI - Comparison of the extracellular proteomes of Escherichia coli B and K-12 strains during high cell density cultivation. AB - Escherichia coli BL21 (DE3) and W3110 strains, belonging to the family B and K 12, respectively, have been most widely employed for recombinant protein production. During the excretory production of recombinant proteins by high cell density cultivation (HCDC) of these strains, other native E. coli proteins were also released. Thus, we analyzed the extracellular proteomes of E. coli BL21 (DE3) and W3110 during HCDC. E. coli BL21 (DE3) released more than twice the amount of protein compared with W3110 during HCDC. A total of 204 protein spots including 83 nonredundant proteins were unambiguously identified by 2-DE and MS. Of these, 32 proteins were conserved in the two strains, while 20 and 33 strain specific proteins were identified for E. coli BL21 (DE3) and W3110, respectively. More than 70% of identified proteins were found to be of periplasmic origin. The outer membrane proteins, OmpA and OmpF, were most abundant. Two strains showed much different patterns in their released proteins. Also, cell density-dependent variations in the released proteins were observed in both strains. These findings summarized as reference proteome maps will be useful for studying protein release in further detail, and provide new strategies for enhanced excretory production of recombinant proteins. PMID- 18425733 TI - A systematic approach to identify STRE-binding proteins of the gsn glycogen synthase gene promoter in Neurospora crassa. AB - The gene encoding glycogen synthase in Neurospora crassa (gsn) is transcriptionally down-regulated when mycelium is exposed to a heat shock from 30 to 45 degrees C. The gsn promoter has one stress response element (STRE) motif that is specifically bound by heat shock activated nuclear proteins. In this work, we used biochemical approaches together with mass spectrometric analysis to identify the proteins that bind to the STRE motif and could participate in the gsn transcription regulation during heat shock. Crude nuclear extract of heat shocked mycelium was prepared and fractionated by affinity chromatography. The fractions exhibiting DNA-binding activity were identified by electrophoretic mobility shift assay (EMSA) using as probe a DNA fragment containing the STRE motif. DNA-protein binding activity was confirmed by Southwestern analysis. The molecular mass (MM) of proteins was estimated by fractionating the crude nuclear extract by SDS-PAGE followed by EMSA analysis of the proteins corresponding to different MM intervals. Binding activity was detected at the 30-50 MM kDa interval. Fractionation of the crude nuclear proteins by IEF followed by EMSA analysis led to the identification of two active fractions belonging to the pIs intervals 3.54-4.08 and 6.77-7.31. The proteins comprising the MM and pI intervals previously identified were excised from a 2-DE gel, and subjected to mass spectrometric analysis (MALDI-TOF/TOF) after tryptic digestion. The proteins were identified by search against the MIPS and MIT N. crassa databases and five promising candidates were identified. Their structural characteristics and putative roles in the gsn transcription regulation are discussed. PMID- 18425734 TI - A short peptide is a protein kinase C (PKC) alpha-specific substrate. AB - The purpose of this study was to find protein kinase C (PKC) isozyme-specific peptides. A peptide library containing 1772 sequences was designed using Scansite and screened by MALDI-TOF MS and kinase activity assays for PKC isozyme specificity. A peptide (Alphatomega; H-FKKQGSFAKKK-NH(2)) with high specificity for PKC alpha relative to other isozymes was identified. The peptide was phosphorylated to a greater extent by tissue lysates from B16 melanoma, HepG2, and human breast cancer, which had higher levels of activated PKC alpha, when compared to normal skin, liver, and human breast tissue lysates, respectively. Moreover, addition of Ro-31-7549, an inhibitor with great specificity for PKC alpha, to the phosphorylation reaction caused a dose-dependent reduction in phosphorylation, but no inhibition was identified with the addition of rottlerin and H-89. These results show that this peptide has great potential as a PKC alpha specific substrate. PMID- 18425735 TI - Structural effects of solvents on the breathing of metal-organic frameworks: an in situ diffraction study. PMID- 18425736 TI - Hydroxycruciforms: amine-responsive fluorophores. AB - The synthesis of three hydroxy-substituted cruciforms (XF, 1,4-bis(4' hydroxystyryl)-2,5-bis(4''-methoxyphenylethynyl)benzene, 1,4-bis(4' methoxystyryl)-2,5-bis(4''-hydroxyphenylethynyl)benzene, and 1,4-bis(4' hydroxystyryl)-2,5-bis(4''-hydroxyphenylethynyl)benzene) starts with a Horner reaction followed by a Sonogashira coupling and subsequent deprotection. The three herein described XFs contain either two or four free phenolic hydroxyl groups. All three XFs were subjected to photometric UV/Vis titrations in a methanol/water mixture. The respective pK(a) values were obtained by data deconvolution. As the three XFs display a significant change in emission color upon photoinduced deprotonation, the XFs were taken up in different solvents and exposed to twelve amines. The amine-dependent change in emissivity of the tetrahydroxy XF is sufficiently distinct in the eight solvents that all of the inspected amines are discerned by a linear discriminant analysis. The tetrahydroxy XF in different solvents forms a sensor array, the response of which is based on the excited-state proton transfer (ESPT) to amines and mediated by the choice of the battery of solvents that are utilized. PMID- 18425737 TI - Broadband dipolar recoupling for magnetization transfer in solid-state NMR correlation spectroscopy. PMID- 18425739 TI - Synthesis and structure of monomeric, trimeric, and mixed phenylcyanamides. AB - In a new synthetic approach phenylcyanamide (Hpca) was synthesized by methylation of phenylthiourea followed by a basic work-up. All products along the synthetic route have been fully characterized by means of NMR, IR, and X-ray studies. The first structural report of neutral mixed crystals of phenylcyanamide containing monomeric and trimeric Hpca is presented. Examination of these intriguing mixed crystals revealed the formation of distinct layers of monomeric and trimeric Hpca. These layers are interconnected by weak hydrogen bonds. The trimer represents triphenylisomelamine, which readily isomerizes to the triphenylmelamine in the melt, in accord with computations at the B3LYP level, indicating an exothermic process (DeltaH = -49.4 kcal mol(-1)). Pure trimeric Hpca (triphenylisomelamine) was obtained either by recrystallization of the mixed crystals from boiling water or by trimerization of monomeric Hpca in isopropanol for 12 h under reflux conditions. For comparison tritylcyanamide (Htca) and potassium phenylcyanamide as an [18]crown-6 complex [K([18]crown-6)pca] have been synthesized, and the solid-state structures were determined using X-ray diffraction techniques. The thermal behavior was studied by thermo-analytical experiments. In agreement with the experimental results, computations predict an exothermic cyclotrimerization process for Hpca (DeltaH = -41.3 kcal mol(-1)). PMID- 18425738 TI - Cellular uptake, cytotoxicity, and metabolic profiling of human cancer cells treated with ruthenium(II) polypyridyl complexes [Ru(bpy)2(N--N)]Cl2 with N- N=bpy, phen, dpq, dppz, and dppn. AB - A series of five ruthenium(II) polypyridyl complexes [Ru(bpy)2(N--N)]Cl2 was tested against human HT-29 and MCF-7 cancer cell lines. Cellular uptake efficiency and cytotoxicity were found to increase with the size of the aromatic surface area of the N--N ligand. The most active compound carrying the dppn ligand exhibits a low micromolar IC(50) value against both cell lines comparable to that of cisplatin under similar conditions. Continuous measurement of oxygen consumption, extracellular acidification rate, and impedance of the cell layer with a chip-based sensor system upon exposure to the complexes showed only small changes for the first two parameters throughout the series. A significant and irreversible decrease in impedance was, however, found for the dppn compound. This suggests that its biological activity is related to modifications in cell morphology or cell-cell and cell-matrix contacts. PMID- 18425740 TI - Novel single-crystal-to-single-crystal anion exchange and self-assembly of luminescent d(10) metal (Cd(II), Zn(II), and Cu(I)) complexes containing C(3) symmetrical ligands. AB - Ligands L1 and L2' (L1=N,N',N''-tris(4-pyridyl)trimesic amide, L2'=N,N',N'' tris(3-pyridinyl)-1,3,5-benzenetricarboxamide) belonging to an interesting family of tripyridyltriamides with C(3)-symmetry have been utilized to construct 3D porous or hydrogen-bonded frameworks. Through a novel single-crystal-to-single crystal anion-exchange process, [Cd(L1)(2)(ClO(4))(2)](n) (1c) can be obtained from [Cd(L1)(2)Cl(2)](n) (1b) in the presence of ClO(4)(-) anions. This anion exchange process is highly selective and only the substitution of Cl(-) by ClO(4)(-) or PF(6)(-) could be realized; Cl(-) was found not to be substituted by BPh(4)(-). This demonstrates that the exchange process depends on the size of the anions in relation to the size of the cavities in the host material (ca. 7.5 A). In addition, the anion-exchange properties of 1 b have also been investigated by means of powder X-ray diffraction (PXRD), elemental analysis (EA), and infrared absorption spectroscopy (IR). Structurally, [Zn(L1)(NO(3))(2)](n)(2) consists of a 2D coordination network with five-coordinate Zn(II) ions. Surprisingly, different trigonal-bipyramidal Zn(II) ions propagate to form distinct respective sheet structures, A and B, which are packed in an A-B-A-B manner in the crystal lattice, and these are hydrogen-bonded to give a 3D extended framework. The molecular structure of [CuI(L2')](n)(3) shows that the Cu(I) ion adopts a distorted tetrahedral geometry, and 3 also forms a 2D coordination network. Significantly, this 2D coordination network is further assembled into a remarkable 3D homochiral framework through triple hydrogen bonding and pi...pi interactions. All of these 3D coordination polymers and/or hydrogen-bonded frameworks are luminescent in the solid state, and their solid-state luminescent properties have been investigated at room temperature and/or at 77 K. PMID- 18425741 TI - New mono- and dimeric members of the secalonic acid family: blennolides A-G isolated from the fungus Blennoria sp. AB - Blennolides A-G (2-8), seven unusual chromanones, were isolated together with secalonic acid B (1) from Blennoria sp., an endophytic fungus from Carpobrotus edulis. This is the first reported isolation of the blennolides 2 and 3 (hemisecalonic acids B and E), the existence of which as the monomeric units of the dimeric secalonic acids had long been postulated. A compound of the proposed structure 4 (beta-diversonolic ester) will need to be revised, as its reported data do not fit those of the established structure of blennolide C (4). Other monomers, the blennolides D-F (5-7) seem to be derived from blennolides A (2) and B (3) by rearrangement of the hydroaromatic ring. The heterodimer 8, composed of the monomeric blennolide A (2) and the rearranged 11-dehydroxy derivative of blennolide E (6), extends the ergochrome family with an ergoxanthin type of skeleton. The structures of the new compounds were elucidated by detailed spectroscopic analysis and further confirmed by an X-ray diffraction study of a single crystal of 2. The absolute configurations were determined by TDDFT calculations of CD spectra, including the solid-state CD/TDDFT approach. Preliminary studies showed strong antifungal and antibacterial activities of these compounds against Microbotryum violaceum and Bacillus megaterium, respectively. They were also active against the alga Chlorella fusca and the bacterium Escherichia coli. PMID- 18425742 TI - Phosphorescent iridium(III) complexes with nonconjugated cyclometalated ligands. AB - A series of blue phosphorescent iridium(III) complexes 1-4 with nonconjugated N benzylpyrazole ligands were synthesized and their structural, electrochemical, and photophysical properties were investigated. Complexes 1-4 exhibit phosphorescence with yields of 5-45 % in degassed CH2Cl2. Of the compounds, 1 showed emission that was nearly true blue at 460 nm with a lack of vibronic progression. These photophysical data clearly demonstrate that the methylene spacer of the cyclometalated N-benzylpyrazole chelate effectively interrupts the pi conjugation upon reacting with a third L X chelating chromophore. This gives a feasible synthesis for the blue phosphorescent complexes with a sufficiently large energy gap. In another approach, these complexes were investigated for their suitability for the host material in phosphorescent OLEDs. The device was synthesized by using 1 as the host for the green-emitting [Ir(ppy)3] dopant, which exhibits an external quantum conversion efficiency (EQE) of up to 11.4 % photons per electron (and 36.6 cdA(-1)), with 1931 Commission Internationale de L'Eclairage (CIE) coordinates of (0.30, 0.59), a peak power efficiency of 21.7 lmW(-1), and a maximum brightness of 32000 cdm(-2) at 14.5 V. At the practical brightness of 100 cdm(-2), the efficiency remains above 11 % and 18 lmW(-1), demonstrating its great potential as the host material for phosphorescent organic light-emitting diodes. PMID- 18425743 TI - Similar biological activities of two isostructural ruthenium and osmium complexes. AB - In this study, we probe and verify the concept of designing unreactive bioactive metal complexes, in which the metal possesses a purely structural function, by investigating the consequences of replacing ruthenium in a bioactive half sandwich kinase inhibitor scaffold by its heavier congener osmium. The two isostructural complexes are compared with respect to their anticancer properties in 1205 Lu melanoma cells, activation of the Wnt signaling pathway, IC(50) values against the protein kinases GSK-3beta and Pim-1, and binding modes to the protein kinase Pim-1 by protein crystallography. It was found that the two congeners display almost indistinguishable biological activities, which can be explained by their nearly identical three-dimensional structures and their identical mode of action as protein kinase inhibitors. This is a unique example in which the replacement of a metal in an anticancer scaffold by its heavier homologue does not alter its biological activity. PMID- 18425744 TI - Gliotoxin enhances radiotherapy via inhibition of radiation-induced GADD45a, p38, and NFkappaB activation. AB - The purpose of the study was to elucidate the mechanism underlying the enhancement of radiosensitivity to 60Co gamma-irradiation in human hepatoma cell line HepG2 pretreated with gliotoxin. Enhancement of radiotherapy by gliotoxin was investigated in vitro with human hepatoma HepG2 cell line. Apoptosis related proteins were evaluated by Western blotting. Annexin V/PI and reactive oxygen species (ROS) were quantified by Flow Cytometric (FACS) analysis. Gliotoxin (200 ng/ml) combined with radiation (4 Gy) treated cells induced apoptosis. Cells treated with gliotoxin (200 ng/ml) prior to irradiation at 4 Gy induced the expression of bax and nitric oxide (NO). The gliotoxin-irradiated cells also increased caspase-3 activation and ROS. Gadd45a, p38, and nuclear factor kappa B (NFkappaB) activated in irradiated cells was inhibited by Gliotoxin. Specific inhibitors of p38 kinase, SB203580, significantly inhibited NFkappaB activation and increased the cytotoxicity effect in cells exposed to gliotoxin combined with irradiation. However, SB203580 did not suppress the activation of Gadd45a in irradiated cells. Gliotoxin inhibited anti-apoptotic signal pathway involving the activation of Gadd45a-p38-NFkappaB mediated survival pathway that prevent radiation-induced cell death. Therefore, gliotoxin, blocking inflammation pathway and enhancing irradiation-induced apoptosis, is a promising agent to increase the radiotherapy of tumor cells. PMID- 18425745 TI - p21 is required for atRA-mediated growth inhibition of MEPM cells, which involves RAR. AB - All-trans retinoic acid (atRA), a metabolite of vitamin A, is essential for embryonic development. Thus the spatial and temporal dispersal of RA must be tightly controlled. Previous studies show that excessive atRA led to growth inhibition and p21 accumulation in mouse embryonic palatal mesenchymal (MEPM) cells. We reported here the identification of p21 as a required mediator during atRA-induced growth inhibition. atRA caused a G1 arrest in the cell cycle with an increase in the proportion of cells in G0/G1 and a decrease in the proportion of cells in S phase. In addition to a marked effect on cell cycling, atRA also triggered DNA fragmentation, reflected by an increase of the fraction of cells in the sub-G(1) population. Western blot analysis revealed that atRA treatment led to an increase in p21 level and a decrease in cyclin D1 protein and Rb phosphorylation. Using luciferase assay with reporter gene regulated by p21 promoter, we showed that atRA increased the reporter activity in a dose-dependent manner; and p21 siRNA blocked the growth inhibition by atRA, suggesting that p21 is required for atRA-mediated growth inhibition. Moreover, the induction of p21 by atRA was partially attenuated when RAR was silenced with specific siRNA. atRA stimulated RARE-driven reporter gene activity dose-dependently. Using chromatin immunoprecipitation, we demonstrated that RAR protein could bind to the p21 promoter. Taken together, our results indicate p21 is responsible for atRA induced growth inhibition of MEPM cells and RAR plays a role during this process. PMID- 18425746 TI - Packed-column capillary electrochromatography and capillary electrochromatography mass spectrometry using a lithocholic acid stationary phase. AB - The preparation and characterization of a novel lithocholic acid (LCA)-based liquid crystalline (LC) stationary phase (SP) suitable for application in packed column CEC and CEC coupled to MS is described. The extent of bonding reactions of LCA-SP was assessed using 1H-NMR, 13C-NMR and elemental analysis. This characterization is followed by application of the LCA-SP for separation of beta blockers, phenylethylamines (PEAs), polyaromatic hydrocarbons (PAHs) and polychlorinated biphenyls (PCBs). Using the optimum mobile phase operating conditions (pH 3.0-4.5, 10 mM ammonium acetate, 85% v/v ACN), a comparison of the chromatographic ability of the aminopropyl silica phase vs. the LCA-bonded phase was conducted. The results showed improved selectivity for all test analytes using the latter phase. For example, the CEC-MS of beta-blockers demonstrated that the LCA-bonded phase provides separation of six out of seven beta-blockers, whereas the amino silica phase provides four peaks of several co-eluting beta blockers. For the CEC-MS analysis of PEAs, the LCA-bonded phase showed improved resolution and different selectivity as compared to the aminopropyl phase. An evaluation of the retention trends for PEAs on both phases suggested that the PEAs were retained based on varying degree of hydroxyl substitution on the aromatic ring. In addition, the MS characterization shows several PEAs fragment in the electrospray either by loss of an alkyl group and/or by loss of H2O. Finally, the LCA-bonded phase displayed significantly higher separation selectivity for PAHs and PCBs as compared to the amino silica phase. PMID- 18425747 TI - CE-MSn of complex pectin-derived oligomers. AB - As pectin molecules are too large and heterogeneous to analyze as a whole, the polymer is usually degraded to smaller oligomers, which are often analyzed by high-performance anion exchange chromatography (HPAEC). However, the high salt concentration necessary to elute pectin oligomers by HPAEC is incompatible with online mass detection. To overcome such a disadvantage, a CE-IT-MS system was set up to further elucidate the fine structure of charged oligosaccharides. An effective separation of differently substituted galacturonic acid containing oligomers was obtained by low-pH CE-LIF analysis. By adapting the buffer and capillary online MS detection was enabled. Moreover, with MS/MS it was possible to localize sugar residues' substitutions. With this combined CE-MS approach LIF electropherograms of xylogalacturonan and rhamnogalacturonan I digests could be annotated. The method was further exemplified by a complex oligomer mixture of acid hydrolyzed apple pectin, which was separated and characterized by CE-MSn. Oligomers present in low amounts could be localized by their corresponding m/z, as was demonstrated by selected mass range representation. PMID- 18425748 TI - Development of a capillary electrophoresis-mass spectrometry method using polymer capillaries for metabolomic analysis of yeast. AB - Metabolomics is an emerging field in analytical biochemistry, and the development of such a method for comprehensive and quantitative analysis of organic acids, carbohydrates, and nucleotides is a necessity in the era of functional genomics. When a concentrated yeast extract was analyzed by CE-MS using a successive multiple ionic-polymer layer (SMIL)-coated capillary, the adsorption of the contaminants on the capillary wall caused severe problems such as no elution, band-broadening, and asymmetric peaks. Therefore, an analytical method for the analysis of anionic metabolites in yeast was developed by pressure-assisted CE using an inert polymer capillary made from poly(ether etherketone) (PEEK) and PTFE. We preferred to use the PEEK over the PTFE capillary in CE-MS due to the easy-to-use PEEK capillary and its high durability. The separation of anionic metabolites was successfully achieved with ammonium hydrogencarbonate/formate buffer (pH 6.0) as the electrolyte solution. The use of 2-propanol washing after every electrophoresis run not only eliminated wall-adsorption phenomena, but allowed for good repeatability to be obtained for migration times in the metabolomic analysis. PMID- 18425749 TI - Characterization of Atropa belladonna L. compounds by capillary electrophoresis electrospray ionization-time of flight-mass spectrometry and capillary electrophoresis-electrospray ionization-ion trap-mass spectrometry. AB - This short communication describes the characterization of seven tropane alkaloid compounds in Atropa belladonna L. Thus a rapid and easy CE-electrospray interface (ESI)-TOF-MS procedure is developed to analyze these compounds in a pharmaceutical preparations of A. belladonna L. leaf extract. Optimum electrophoretic separation is obtained using an alkaline solution of 60 mM ammonium acetate at pH 8.5 containing 5% isopropanol. Under the optimum CE-ESI TOF-MS conditions several important compounds such as tropine, belladonnine, norhyoscyamine, apoatropine, hyoscyamine, 6beta-hydroxyhyoscyamine, and scopolamine have been simultaneously identified from A. belladonna L. CE-ESI-IT MS has been used to discriminate the putative presence of littorine. The sensitivity, together with mass accuracy and true isotopic pattern of the TOF-MS, allowed the identification of a broad series of tropane alkaloid compounds present in pharmaceutical preparations of A. belladonna L. leaf extract. PMID- 18425751 TI - Characterization of intact lipopolysaccharides from the Haemophilus influenzae strain RM 118 using electrophoresis-assisted open-tubular liquid chromatography mass spectrometry. AB - We have applied an electrophoresis-assisted open-tubular LC-MS method for analyzing intact lipopolysaccharides (LPSs) from Haemophilus influenzae strain RM118 (Rd). We were able to obtain structural information on both core oligosaccharides (OSs) and the lipid A moiety including the sialylation, glycylation, and the distribution of fatty acid residues on the disaccharide backbone of lipid A. The fragmentation patterns of sodiated and protonated LPS molecules were investigated for determining the location of sialic acid. It was found that the tandem mass spectra of sodiated ions provided unambiguous evidence of both sialylated lactose and sialylated lacto-N-neotetraose. In contrast, the fragment ions of protonated ions only offered the evidence for the existence of sialylated lacto-N-neotetraose. The lipid A of Gram-negative bacteria, as the principal endotoxic component of LPS, plays a major role in the pathogenesis of bacterial infections. We have previously characterized lipid A species after mild acid hydrolysis of LPS during which lipid A precipitates. In this study, intact LPS was directly introduced to a tandem mass spectrometer. In-source dissociation strategy was employed, followed by multiple-stage MS/MS on the ions originating from the lipid part to obtain structural information. This is the first time that the structure of lipid A of H. influenzae was characterized by MS/MS on intact LPS molecules without any prior chemical modifications. In the same way information on the OS can be obtained by MS/MS by focusing on ions originating from core OS. PMID- 18425750 TI - Application of capillary isotachophoresis-based multidimensional separations coupled with electrospray ionization-tandem mass spectrometry for characterization of mouse brain mitochondrial proteome. AB - By employing a capillary ITP (CITP)/CZE-based proteomic technology, a total of 1795 distinct mouse Swiss-Prot protein entries (or 1705 nonredundant proteins) are identified from synaptic mitochondria isolated from mouse brain. The ultrahigh resolving power of CITP/CZE is evidenced by the large number of distinct peptide identifications measured from each CITP fraction together with the low peptide fraction overlapping among identified peptides. The degree of peptide overlapping among CITP fractions is even lower than that achieved using combined CIEF/nano-RP LC separations for the analysis of the same mitochondrial sample. When evaluating the protein sequence coverage by the number of distinct peptides mapping to each mitochondrial protein identification, CITP/CZE similarly achieves superior performance with 1041 proteins (58%) having 3 or more distinct peptides, 233 (13%) having 2 distinct peptides, and 521 (29%) having a single distinct peptide. The reproducibility of protein identifications is found to be around 86% by comparing proteins identified from repeated runs of the same mitochondrial sample. The analysis of the mouse mitochondrial proteome by two CITP/CZE runs results in the detection of 2095 distinct mouse Swiss-Prot protein entries (or 1992 nonredundant proteins), corresponding to 59% coverage of the updated Maestro mitochondrial reference set. The collective analysis from combined CITP/CZE and CIEF-based proteomic studies yields the identification of 2191 distinct mitochondrial protein entries (or 2082 nonredundant proteins), corresponding to 76% coverage of the MitoP2-database reference set. PMID- 18425752 TI - Analysis of intact erythropoietin and novel erythropoiesis-stimulating protein by capillary electrophoresis-electrospray-ion trap mass spectrometry. AB - Recombinant human erythropoietin (rHuEPO) and novel erythropoiesis-stimulating protein (NESP) were analyzed by CE-ESI-MS using an IT as analyzer. The IT parameters were optimized by direct infusion of solutions of different intact proteins (myoglobin, transferrin, alpha1-acid glycoprotein and fetuin) with different degrees of glycosylation (from 0 to 35% w/w). Two physically adsorbed capillary coatings from UltraTol Pre-Coats (low normal (LN) and high reverse (HR)) were evaluated for the separation of rHuEPO and NESP glycoforms by CE-ESI IT-MS. The results obtained with the neutral LN coating suggest that an IT mass spectrometer enables identification of the main glycoforms of a complex glycoprotein such as rHuEPO. Although LN provided acceptable glycoform resolution for rHuEPO, the separation obtained for NESP was less significant due to the higher microheterogeneity of this glycoprotein. Reproducibility studies confirmed the lack of stability and bleeding of the LN coating, which caused problems with MS detection, such as a dramatic loss of sensitivity and the presence of peaks in the mass spectra corresponding to molecular ions in the coating. In contrast, the cationic HR coating gave faster but poorer glycoform separations due to the presence of an anodal EOF. However, the positive charge of the coating provided enhanced hydrolytic stability, making it more suitable than the LN coating for the on-line MS coupling. PMID- 18425753 TI - Analysis of major alkaloids in Rhizoma coptidis by capillary electrophoresis electrospray-time of flight mass spectrometry with different background electrolytes. AB - CE-based techniques with DAD and detection ESI-TOF-MS have been developed for the analysis of seven protoberberine alkaloids and one aporphinoid alkaloid in Huanglian (Rhizoma coptidis), a well-known traditional Chinese herbal medicine. One aqueous BGE and one nonaqueous BGE were developed for CE-DAD and CE-MS analyses, and the CE-ESI-TOF-MS conditions including nebulizer gas pressure, the sheath-liquid composition, its flow rate, etc. were optimized. Eight main alkaloids in R. coptidis could be separated with baseline resolution by CE-DAD with these two different BGEs, and identified by TOF-MS analysis. Moreover, three major alkaloids (berberine, palmatine, and jatrorrhizine) could be quantified accurately by CE-DAD and CE-MS with the BGE system consisting of 50:50 v/v water and ACN containing 50 mM ammonium acetate at pH 6.8. Both techniques provided similar LODs and could be applied with confidence within similar linear dynamic range. However, reproducibility and speed of analysis were better using CE-DAD. When the CE technique was compared with the RP-HPLC method, the CE-DAD and CE-MS methods provided greater efficiency and faster analysis speed, i.e., achieving baseline resolution for all the eight main basic compounds in less than 14 min. The CE method, as a viable alternative to HPLC, is suitable for use as a routine procedure for the rapid identification and quantification of basic compounds in herbal or natural product applications. PMID- 18425754 TI - Determination of sialic acids released from glycoproteins using capillary zone electrophoresis/electrospray ionization mass spectrometry. AB - A simple method for the determination of the two most abundant sialic acids released from glycoproteins based on CZE-MS is presented. Several parameters like BGE with various organic modifiers and sheath liquids were studied with respect to their suitability for the fast and easy analysis of the selected compounds by CZE-MS. Finally, a BGE containing 10 mM ammonium acetate allowed the quantification of N-acetylneuraminic acid (Neu5Ac) and N-glycolylneuraminic acid (Neu5Gc) in glycoproteins as well as human plasma samples. LOD and LOQ were 2 microg/mL and 6 microg/mL, respectively. PMID- 18425755 TI - Capillary isoelectric focusing hyphenated to single- and multistage matrix assisted laser desorption/ionization-mass spectrometry using automated sheath flow-assisted sample deposition. AB - In this paper CIEF combined with MALDI-MS is described using a sheath-liquid assisted automatic sample deposition from the separation capillary onto a MALDI target. Sample/matrix preparation techniques on the target resembling the dried droplet and the thin layer methods were evaluated in the context of the automatic spotting. Volatile buffers were used as IEF catholyte solutions. Test samples consisting of tryptic peptides, glycopeptides, and phosphopeptides of well-known proteins showed that CIEF-MALDI-MS can be used as effective preseparation method prior to MS, allowing to obtain the amino acid sequence coverage of proteins similar to that achieved with CZE-MALDI-MS and CZE-ESI-MS. Particularly, completeness and reliability of glycopeptide analysis is much enhanced by the preseparation. The effect is less pronounced but still significantly found with phosphopeptides present in the test protein. Finally, a test sample of five standard proteins demonstrates the suitability of this technique also for the treatment of intact proteins. This technique has potential to emerge as a faster method analogous and complementary to 2-DE and to IPG-IEF-MALDI-MS demonstrated before by the group of Loo [1]. PMID- 18425756 TI - Nonaqueous capillary electrophoresis-electrospray ionization-ion trap-mass spectrometry analysis of pyrrolo- and pyrido[1,2-a]azepine alkaloids in Stemona. AB - Stemona alkaloids represent an outstanding class of natural compounds due to their pharmacological profile and their complex and unusual molecular structures. The aim of this study was the development of the first CE method for the separation, identification and quantification of these pyrrolo- and pyrido[1,2 a]azepine derivatives in three Stemona species. The best results were obtained with a NACE-ESI-IT-MS method, utilizing an electrolyte of 50 mM ammonium acetate, 1 M acetic acid and 10% methanol in ACN and a separation voltage of 30 kV. Samples were injected voltage-assisted with 20 kV for 1 s. Isopropanol:water (1:1) was used as ESI sheath liquid at a flow rate of 3 microL/min. The assay was applied for the qualitative profiling of Stemona alkaloids in S. curtisii, S. collinsae and S. tuberosa. For unambiguous peak assignment of more than forty unidentified alkaloids MS/MS experiments were performed and fragmentation patterns studied. Subsequently the method was validated for the quantitative determination of four selected derivatives (RSD inter- and intraday <6%, LODs <7.5 microg/mL, LOQs <25.0 microg/mL, for all analytes, recovery rates >98.9%) in several Stemona sp. extracts. PMID- 18425757 TI - A new insight into suction and dilution effects in capillary electrophoresis coupled to mass spectrometry via an electrospray ionization interface. Part I Suction effect. AB - The hyphenation of CE with MS is nowadays accepted as a powerful analytical approach. Employing ESI, the most common interface, one challenge is to provide quantitative information, which is quite a difficult task, as it is linked, among other factors, to suction and dilution effects. In the coaxial ESI configuration, the suction effect has been presented in literature as stemming from nebulizing gas (NG) flow rate and drying gas temperature. But as this interface consists in three concentric capillaries, allowing for BGE, sheath liquid (SL) and NG mixing, it is demonstrated herein that other parameters are also involved in this suction effect: the CE capillary protrusion from the interface needle, SL flow rate, and overall BGE flow rate and velocity profile. Whereas NG flow rate is the parameter affecting suction to a greater extent, separation capillary protruding length, SL, and overall BGE flow rate have a significant additional impact on this phenomenon. It is shown that SL flow rate can affect suction differently according to the NG velocity, which may be explained by modification of the Taylor cone geometry. Furthermore, it appears that suction effect is noticeably favored by a parabolic velocity profile of the BGE, again probably due to the Taylor cone shape modification. Finally, the temperature gradient created by the contact between the heated NG and the separation capillary enhances this effect. PMID- 18425758 TI - Identification of toxic oligopeptides in Amanita fungi employing capillary electrophoresis-electrospray ionization-mass spectrometry with positive and negative ion detection. AB - The identification of toxic oligopeptides employing CE-ESI-MS is presented. The analytes studied ama- and phallotoxins are of significant forensic interest because over 90% of the lethal cases of fungus poisoning in man are caused by species of Amanita which contain these toxins. A CE method was developed to separate the toxins alpha-, beta- and gamma-amanitin, phalloidin and phallacidin. Their fragmentation patterns in MS(n) experiments were investigated in the positive and in the negative ion mode, also the influence of the sheath liquid mixture of the used interface on the S/N. Method validation included the determination of the LOD and the repeatability of the migration time and peak area for both detection modes. With the optimized method LODs of 13-79 ng/mL (17 87 nmol/L) were reached. The CE-MS procedure was successfully applied to the identification of ama- and phallotoxins in extracts of air-dried mushroom samples. PMID- 18425759 TI - Growth and manipulation of yeast. AB - Yeast cultures can be grown, maintained, and stored in liquid media or on agar plates using techniques similar to those for bacterial cultures. This unit describes culture conditions for these basic techniques. Additional methods describe determination of yeast mating type, diploid construction, sporulation, tetrad dissection, and random spore analysis. PMID- 18425760 TI - EMS and UV mutagenesis in yeast. AB - Many fundamental biological processes have been elucidated by the isolation and analysis of mutants that are defective in such processes. Therefore, the methods to generate mutants are of great importance in model organisms. This unit describes two protocols for mutagenesis of yeast-using ethyl methanesulfate (EMS) and ultraviolet (UV) light. Each of these methods has been used successfully for many years. PMID- 18425761 TI - Using CellProfiler for automatic identification and measurement of biological objects in images. AB - Visual analysis is required to perform many biological experiments, from counting yeast colonies to measuring the size and shape of individual cells or the intensity of fluorescently labeled proteins within them. This unit outlines the use of CellProfiler, a free, open-source image analysis tool that extracts quantitative information from biological images. It includes a step-by-step protocol for automated analysis of the number, color, and size of yeast colonies growing on agar plates, but the methods can be adapted to identify and measure any objects in images. The flexibility of the software allows users to tailor pipelines of adjustable modules to fit different biological experiments, to generate accurate measurements from dozens or even hundreds of thousands of images. PMID- 18425762 TI - In situ polymerase chain reaction and hybridization to detect low-abundance nucleic acid targets. AB - This unit provides detailed methods and material descriptions for in situ hybridization following in situ amplification of DNA or RNA by PCR. It includes all essential components of the techniques, including variations suitable for different kinds of tissue and cell preparations. Planning, controls, and critical parameters for the amplification steps are discussed. PMID- 18425763 TI - Interaction trap/two-hybrid system to identify interacting proteins. AB - The yeast two-hybrid method (or interaction trap) is a powerful technique for detecting protein interactions. The procedure is performed using transcriptional activation of a dual reporter system in yeast to identify interactions between a protein of interest (the bait protein) and the candidate proteins for interaction. The method can be used to screen a protein library for interactions with a bait protein or to test for association between proteins that are expected to interact based on prior evidence. Interaction mating facilitates the screening of a library with multiple bait proteins. PMID- 18425764 TI - Production of a heterozygous mutant cell line by homologous recombination (single knockout). AB - Gene targeting by homologous recombination is a powerful and widely used technique for introduction of specific gene mutations (frequently a gene inactivation) in transgenic animals. The basic method detailed in this unit uses sequences homologous to the endogenous gene flanking the mutation. While methods using bacterial artificial chromosomes (BACs) and recombineering may be used, in most cases simpler bacterial plasmid clones with several kb of homology are sufficient. This protocol details the strategic factors in designing the constructs for selection and screening for homologous recombination. PMID- 18425765 TI - Production of a homozygous mutant embryonic stem cell line (double knockout). AB - Under some circumstances, it may be desirable to produce a mouse cell clone in which both alleles of a desired gene are mutated. This may be because the mutation causes embryonic lethality in homozygous animals, or to test cultured cells before an animal is produced. This protocol details an easy method for obtaining homozygous cells by homologous recombination without the need for two targeting events. PMID- 18425766 TI - PRNP mutations in a series of apparently sporadic neurodegenerative dementias in China. AB - Mutations in prion protein gene (PRNP) may lead to genetic prion disease, which usually has a broad range of phenotypic presentations that overlap with other neurodegenerative dementias. In this study, we screened the PRNP gene to evaluate the frequency of PRNP mutations and their correlations with clinical phenotype in 185 sporadic neurodegenerative dementia cases and 310 control subjects. Samples of DNA from each subject underwent polymerase chain reaction (PCR) amplification and direct sequencing of PRNP. The clinical characteristics of patients carrying PRNP mutations were detailed. We identified five different PRNP mutations in five patients, of which three were novel (S97N, F198V, and R208C) and two were known (D178N-129M and M232R). The rate of PRNP mutation was 2.70% in our sample. Though future studies confirming the correlation between PRNP mutations and clinical phenotype need to be undertaken, PRNP genotyping may be a valuable tool to differentiate between prion disease and other neurodegenerative dementias. PMID- 18425769 TI - Histone deacetylases as transducers and targets of nuclear signaling. AB - Histone deacetylase (HDAC) activity was first discovered about 40 years ago, but it was not until the molecular identification of the first HDACs in 1996 that this family of enzymes gained prominence. In addition to histones, HDACs reverse lysine acetylation of various non-histone proteins located in the nucleus and the cytoplasm. Here, we examine the nuclear roles of these enzymes, with a specific focus on their active crosstalk with different chromatin regulators. PMID- 18425770 TI - AKT as locus of fragility in robust cancer system. AB - Metastatic cancer is a complex positive feedback loop system. Such as system has a tendency to acquire extreme robustness. Signaling pathways controlling that robustness can fail completely if an essential element from the signaling is removed. That element is a locus of fragility. Targeting that locus represents the best way to target the cancer robustness. This prospect presents another locus of fragility in signaling complex system network, controlling the cell cycle progression through the PI3K/AKT/mTOR/RAN pathway and cell migration and angiogenesis through the VEGF/PI3K/AKT/NO/ICAM-1 pathway. The locus of fragility of these pathways is AKT, which is regulated by a balance of catalase/H2O2 or by AKT inhibitor. Tiny and trivial perturbations such as change in redox state in the cells by antioxidant enzyme catalase, scavenging H2O2 signaling molecule, regulates robust signaling molecule AKT, abolishing its phosporilation and inducing cascading failure of robust signaling pathways for cell growth, proliferation, migration, and angiogenesis. An anticancer effect of the antioxidant is achieved through the AKT locus, by abolishing signals from growth factors VEGF, HGF, HIF-1alpha and H2O2. Previously reported locus of fragility nitric oxide (NO) and locus AKT are close in the complex signaling interactome network, but they regulate distinct signaling modules. Simultaneously targeted loci represents new principles in cancer robustness chemotherapy by blocking cell proliferation, migration, angiogenesis and inducing rather slow then fast apoptosis leading to slow eradication of cancer. PMID- 18425771 TI - Centromeres in cell division, evolution, nuclear organization and disease. AB - As the spindle fiber attachment region of the chromosome, the centromere has been investigated in a variety of contexts. Here, we will review current knowledge about this unique chromosomal region and its relevance for proper cell division, speciation, and disease. Understanding the three-dimensional organization of centromeres in normal and tumor cells is just beginning to emerge. Multidisciplinary research will allow for new insights into its normal and aberrant nuclear organization and may allow for new therapeutic interventions that target events linked to centromere function and cell division. PMID- 18425772 TI - Cathepsin L inhibitor I blocks mitotic chromosomes decondensation during cleavage cell cycles of sea urchin embryos. AB - We have previously reported that sperm histones (SpH) degradation after fertilization is catalyzed by a cystein-protease (SpH-protease). Its inhibition blocks the degradation of SpH in vivo and also aborts sea urchin development at the initial embryonic cell cycles. It remains unknown if this effect is a consequence of the persistence of SpH on zygotic chromatin, or if this protease is involved per-se in the progression of the embryonic cell cycles. To discriminate among these two options we have inhibited this protease at a time when male chromatin remodeling was completed and the embryos were engaged in the second cell cycle of the cleavage divisions. The role of this enzyme in cell cycle was initially analyzed by immuno-inhibiting its SpH degrading activity in one of the two blastomeres after the initial cleavage division, while the other blastomere was used as a control. We found that in the blastomere injected with the anti-SpH-protease antibodies the cytokinesis was arrested, the chromatin failed to decondense after mitosis and BrdU incorporation into DNA was blocked. Since the N-terminal sequence and the SpH protease was homologous to the cathepsin L (Cat L) family of proteases, we subsequently investigated if the deleterious effect of the inhibition of this protease is related to its Cat L activity. In this context we analyzed the effect of Cat L inhibitor I (Z-Phe-Phe CH(2)F) on embryonic development. We found that the addition of 100 uM of this inhibitor to the embryos harvested at the time of the initial cleavage division (80 min p.i.) mimics perfectly the effects of the immuno-inhibition of this enzyme obtained by microinjecting the anti-SpH-protease antibodies. Taken together these results indicate that the activity of this protease is required for embryonic cell cycle progression. Interestingly, we observed that when this protease was inhibited the chromatin decondensation after mitosis was abolished indicating that the inhibition of this enzyme affects chromosomes decondensation after mitosis. PMID- 18425774 TI - Methylation changes in porcine primordial germ cells. AB - Epigenetic re-programming is an important event in the development of primordial germ cells (PGC) into functional gametes, characterized by genome-wide erasure of DNA methylation and re-establishment of epigenetic marks, a process essential for restoration of the potential for totipotency. In this study changes in the methylation status of centromeric repeats and two IGF2-H19 differentially methylated domain (DMD) sequences were examined in porcine PGC between Days 24 and 31 of pregnancy. The methylation levels of centromeric repeats and IGF2-H19 DMD sequences decreased rapidly from Days 24 to 28 in both male and female PGC. At Days 30 and 31 of pregnancy centromeric repeats and IGF2-H19 DMD sequences acquired new methylation in male PGC, while in female PGC these sequences were completely demethylated by Day 30 and remained hypomethylated at Day 31. To characterize methylation changes that PGC undergo in culture, the methylation status of embryonic germ cells (EGCs) derived from PGC at Day 26 of pregnancy was examined. Centromeric repeats and IGF2-H19 DMD sequences were similarly methylated in both male and female EGC and hypermethylated in female EGC compared with female PGC at the same embryonic age. Our results show that, similar to murine PGC, porcine PGC undergo genome-wide DNA demethylation shortly after arrival in the genital ridges. When placed in culture porcine PGC terminate their demethylation program and may acquire new DNA methylation marks. To our knowledge, this is the first report regarding epigenetic re-programming of genital ridge PGC in the pig. PMID- 18425773 TI - Netrin-1 can affect morphogenesis and differentiation of the mouse mammary gland. AB - Netrin-1 has been shown to regulate the function of the EGF-like protein Cripto-1 (Cr-1) and affect mammary gland development. Since Cr-1 is a target gene of Nanog and Oct4, we investigated the relationship between Netrin-1 and Cr-1, Nanog and Oct4 during different stages of development in the mouse mammary gland. Results from histological analysis show that exogenous Netrin-1 was able to induce formation of alveolar-like structures within the mammary gland terminal end buds of virgin transgenic Cripto-1 mice and enhance mammary gland alveologenesis in early pregnant FVB/N mice. Results from immunostaining and Western blot analysis show that Netrin-1, Nanog and Oct4 are expressed in the mouse embryonic mammary anlage epithelium while Cripto-1 is predominantly expressed outside this structure in the surrounding mesenchyme. We find that in lactating mammary glands of postnatal FVB/N mice, Netrin-1 expression is highest while Cripto-1 and Nanog levels are lowest indicating that Netrin-1 may perform a role in the mammary gland during lactation. HC-11 mouse mammary epithelial cells stimulated with lactogenic hormones and exogenous soluble Netrin-1 showed increased beta-casein expression as compared to control thus supporting the potential role for Netrin-1 during functional differentiation of mouse mammary epithelial cells. Finally, mouse ES cells treated with exogenous soluble Netrin-1 showed reduced levels of Nanog and Cripto-1 and higher levels of beta-III tubulin during differentiation. These results suggest that Netrin-1 may facilitate functional differentiation of mammary epithelial cells and possibly affect the expression of Nanog and/or Cripto-1 in multipotent cells that may reside in the mammary gland. PMID- 18425775 TI - Heparin-binding proteins of human seminal plasma: purification and characterization. AB - Human seminal plasma (HuSP) contains several proteins that bind heparin and related glycosaminoglycans. Heparin binding proteins (HBPs) from seminal plasma have been shown to participate in modulation of capacitation or acrosome reaction and thus have been correlated with fertility in some species. However, these have not been studied in detail in human. The objective of this study was to purify major HBPs from HuSP in order to characterize these proteins. HBPs were isolated by affinity-chromatography on Heparin-Sepharose column, purified by reverse-phase high-performance liquid chromatography (RP-HPLC) and Size-exclusion chromatography and checked for purity on sodium-dodecyl PAGE (SDS-PAGE). Identification of HBPs was done by matrix-assisted laser desorption-ionization time-of-flight-mass spectrometry (MALDI-TOF-MS). Here we report the purification and identification of seven HBPs in seminal fluid. The major HBPs are lactoferrin and its fragments, semenogelin I fragments, semenogelin II, prostate specific antigen, homolog of bovine seminal plasma-proteins (BSP), zinc finger protein (Znf 169) and fibronectin fragments. In this study we are reporting for the first time the purification and identification of BSP-homolog and Znf 169 from HuSP and classified them as HBPs. Here we report the purification of seven clinically important proteins from human seminal fluid through heparin affinity chromatography and RP-HPLC, in limited steps with higher yield. PMID- 18425776 TI - In vitro and in vivo derived porcine embryos possess similar, but not identical, patterns of Oct4, Nanog, and Sox2 mRNA expression during cleavage development. AB - In vitro culture conditions stress the cleavage stage mammalian embryo and can contribute to reduced developmental potential of cultured embryos. One process that may be altered during embryo culture is the establishment and maintenance of pluripotency. Pluripotency is largely controlled by three genes: Oct4, Nanog, and Sox2. The objective of this study was to determine the expression pattern of Oct4, Nanog, and Sox2 in cleavage stage porcine embryos obtained in vivo or by in vitro fertilization and parthenogenetic activation. We used quantitative, real time PCR to assess the relative amount of each transcript in cleavage stage embryos. We found that Oct4 was transiently activated at the 2-cell stage (P value <0.05) while Nanog and Sox2 were activated at the 4-cell stage (P-value <0.05) in in vitro embryos. Embryos derived in vivo showed a similar but not identical pattern of expression of Nanog mRNA been in highest abundance both at the 4 cell and the blastocyst stage. The activation observed at the 4-cell stage for Nanog and Sox2 was shown to be RNA polymerase II dependent (P-value <0.05). This study showed that Oct4, Nanog, and Sox2 possess similar, but not identical, patterns of expression between in vitro and in vivo derived porcine embryos. The difference between the amount of transcripts may reflect the reduced developmental potential observed in in vitro cultured embryos. PMID- 18425778 TI - Diagnosis and management of desmoid tumors and fibrosarcoma. AB - Fibrous tumors represent a diverse subtype of soft tissue tumors and can represent benign conditions as well as frankly malignant sarcomas. Desmoid tumors and dermatofibrosarcoma protuberans are more difficult to classify and tend to be considered in the intermediate risk category. They are distinct entities, but both are locally aggressive processes which are plagued with attendant morbidity and high recurrence rates. Complete surgical resection is the mainstay of treatment. PMID- 18425777 TI - Expression profile of male germ cell-associated genes in mouse embryonic stem cell cultures treated with all-trans retinoic acid and testosterone. AB - Cells that morphologically and functionally resemble male germ cells can be spontaneously derived from ES cells. However, this process is inefficient and unpredictable suggesting that the expression pattern of male germ cell associated genes during spontaneous ES cell differentiation does not mimic the in vivo profiles of the genes. Thus, in the present study, the temporal profile of genes expressed at different stages of male germ cell development was examined in differentiating ES cells. The effect of all-trans retinoic acid (RA) which is a known inducer of primordial germ cell (PGC) proliferation/survival in vitro and testosterone which is required for spermatogenesis in vivo on the expression of these genes was also determined. Each of the 12 genes analyzed exhibited one of four temporal expression patterns in untreated differentiating ES cells: progressively decreased (Dppa3, Sycp3, Msy2), initially low and then increased (Stra8, Sycp1, Dazl, Act, Prm1), initially decreased and then increased (Piwil2, Tex14), or relatively unchanged (Akap3, Odf2). RA-treated cells exhibited increased expression of Stra8, Dazl, Act, and Prm1 and suppressed expression of Dppa3 compared to untreated controls. Furthermore, testosterone increased expression of Stra8 while the combination of RA and testosterone synergistically increased expression of Act. Our findings establish a comprehensive profile of male germ cell gene expression during spontaneous differentiation of murine ES cells and describe the capacity of RA and testosterone to modulate the expression of these genes. Furthermore, these data represent an important first step in designing a plausible directed differentiation protocol for male germ cells. PMID- 18425779 TI - Surgical treatment of invasive squamous cell carcinoma of the penis: Brazilian National Cancer Institute long-term experience. AB - BACKGROUND AND OBJECTIVES: We reviewed our long-term experience with surgical treatment of patients with penile carcinoma. METHODS: From 1960 to 2006, 688 patients with penile carcinoma underwent surgical treatment at our Institute. Several forms of surgical treatment were compared and follow-up data analyzed. RESULTS: Stage stratification demonstrated a better survival rate for patients with stages T1N0 and T1N1,T2N0-1. Patients with well differentiated carcinoma had a higher survival rate than those with moderately and poorly differentiated carcinoma (P < 0.0001 and P = 0.006). Risk stratification showed a better survival rate for patients in the low-risk group (T1G1,T1G2) (P = 0.013 and P < 0.00001). Patients in the intermediate group (T2G1,T2G2,T3G1,T3G2) presented a higher survival rate than patients in the high-risk group (T1-3G3,T4G1-3) (P < 0.00001). Patients who underwent immediate lymphadenectomy had a better survival rate than those who underwent delayed lymphadenectomy (P = 0.002). CONCLUSIONS: Stage and tumor grade affected the prognosis of the disease. The presence and the extent of metastasis to the inguinal region were the most important prognostic factors for survival in our patients. Immediate lymphadenectomy is indicated in all patients. Since recurrences were noted within 8, 10, and 25 years after primary treatment, a frequent and lasting follow-up is essential for all patients. PMID- 18425780 TI - The mitochondrial gateway to cell death. AB - Mitochondria play a key role in death signaling. The intermembrane space of these organelles contains a number of proteins which promote cell death once they are redistributed to the cytosol. The formation of pores in the outer membrane of mitochondria defines a gateway through which the apoptogenic proteins pass during death signaling. Interactions between pro-apoptotic and pro-survival members of the Bcl-2 family of proteins are decisive in the initiation of pore opening. While the specific composition of the pore in molecular terms is still subject to debate and continuing investigation, it is recognized functionally as a passive channel which not only allows egress of proteins to cytosol but also entry in the reverse direction. A variety of constraints may restrict the release of proteins from the intermembrane space to the cytosol. These include trapping in the intercristal spaces formed by the convoluted invaginations of the inner membrane, binding of proteins to the inner membrane or to other soluble proteins of the intermembrane space, or insertion of proteins into the inner membrane. There is a corresponding variety of mechanisms that facilitate release of apoptogenic proteins from such entrapment. Morphological changes that expand the inner membrane enable proteins to be released from enclosure in intercristal spaces, allowing these proteins access to the mitochondrial gateway. Specific cases include cytochrome c molecules bound to inner membrane cardiolipin and released upon oxidation of that lipid component. Further, AIF that is embedded in the inner membrane is released by proteases (caspases or calpains), which enter from the cytosol once the outer membrane pore has opened. The facilitation (or restriction) of apoptogenic protein release through the mitochondrial gateway may provide new opportunities for regulating cell death. PMID- 18425784 TI - Cryoablative therapy in breast cancer: no. PMID- 18425781 TI - Update of the Pompe disease mutation database with 107 sequence variants and a format for severity rating. AB - Pompe disease was named after the Dutch pathologist Dr JC Pompe who reported about a deceased infant with idiopathic hypertrophy of the heart. The clinical findings were failure to thrive, generalized muscle weakness and cardio respiratory failure. The key pathologic finding was massive storage of glycogen in heart, skeletal muscle and many other tissues. The disease was classified as glycogen storage disease type II and decades later shown to be a lysosomal disorder caused by acid alpha-glucosidase deficiency. The clinical spectrum of Pompe disease appeared much broader than originally recognized. Adults with the same enzyme deficiency, alternatively named acid maltase deficiency, were reported to have slowly progressive skeletal muscle weakness and respiratory problems, but no cardiac involvement. The clinical heterogeneity is largely explained by the kind and severity of mutations in the acid alpha-glucosidase gene (GAA), but secondary factors, as yet unknown, have a substantial impact. The Pompe disease mutation database aims to list all GAA sequence variations and describe their effect. This update with 107 sequence variations (95 being novel) brings the number of published variations to 289, the number of non-pathogenic mutations to 67 and the number of proven pathogenic mutations to 197. Further, this article introduces a tool to rate the various mutations by severity, which will improve understanding of the genotype-phenotype correlation and facilitate the diagnosis and prognosis in Pompe disease. PMID- 18425785 TI - Cryoablation for breast cancer: no need to turn a cold shoulder. PMID- 18425786 TI - Is wedge resection of hepatic colorectal cancer metastases an oncologically sound alternative? PMID- 18425787 TI - Commentary on penile cancer article. PMID- 18425788 TI - Gender differences in genetic linkage and association on 11p15 in obsessive compulsive disorder families. AB - Several clinical and genetic studies have reported gender differences in obsessive-compulsive disorder (OCD). Previously, we conducted a linkage genome scan using multipoint allele-sharing methods to test for linkage in 219 families participating in the OCD Collaborative Genetics Study. When these families were stratified by proband's gender, suggestive linkage to chromosome 11p15 at marker D11S2362 (KAC(all) = 2.92, P = 0.00012) was detected in families with male probands, but not in the ones with female probands. We have since conducted fine mapping with a denser microsatellite marker panel in the region of 11p15, and detected a significant linkage signal at D11S4146 (KAC(all) = 5.08, P < 0.00001) in the families of male probands. Subsequently, 632 SNPs were genotyped spanning a 4.0 Mb region of the 1 LOD unit interval surrounding the linkage peak in the original families and an additional 165 families. Six SNPs were associated with OCD (P < 0.001): two SNPs were identified when all the families were included, and four SNPs only in male proband families. No SNP showed significant association with the OCD phenotype only in the families with a female proband. The results suggest a possible gender effect in the etiology of OCD. PMID- 18425789 TI - Impact of type of liver resection on the outcome of colorectal liver metastases: a case-matched analysis. AB - BACKGROUND: Wedge resection (WR) for colorectal liver metastases (CLM) has become more common in an attempt to preserve liver parenchyma. However, some investigator have reported that WR is associated with a higher incidence of positive margin and an inferior survival compared with anatomic resection (AR) 1. OBJECTIVES: This study evaluated survival, margin status, and pattern of recurrence of patients with CLM treated with WR or AR. METHODS: We identified 208 consecutive patients, in a single institutional database from 1995 to 2004, who underwent either WR or AR. WR was defined as a nonanatomic resection and AR was defined as single resection of one or two liver segments. Patients with combined WR-AR and patients requiring resection of more than two segments or radiofrequency ablation were excluded from the analysis. RESULTS: One hundred six patients underwent WR and 102 patients had AR. There were no differences in the rate of positive surgical margin (P = 0.146), overall recurrence rates (P = 0.211), and patterns of recurrence between the two groups (P = 0.468). The median survival was 32 months for WR and 42 for AR, with 5-year survival rates of 29% and 27% respectively, with no significant difference (P = 0.308). Morbidity was similar between the two groups. CONCLUSIONS: WR is a safe procedure and does not disadvantage the patients in terms of tumor recurrence and overall survival. PMID- 18425790 TI - Diffusion of treatment research: does Open Access matter? AB - Advocates of the Open Access movement claim that removing access barriers will substantially increase the diffusion of academic research. If successful, this movement could play a role in efforts to increase utilization of psychotherapy research by mental health practitioners. In a pair of studies, mental health professionals were given either no citation, a normal citation, a linked citation, or a free access citation and were asked to find and read the cited article. After 1 week, participants read a vignette on the same topic as the article and gave recommendations for an intervention. In both studies, those given the free access citation were more likely to read the article, yet only in one study did free access increase the likelihood of making intervention recommendations consistent with the article. PMID- 18425791 TI - Feminist identity as a predictor of eating disorder diagnostic status. AB - Passive Acceptance (PA) and Active Commitment (AC) subscales of the Feminist Identity Development Scale (FIDS) were examined as predictors of eating disorder diagnostic status as assessed by the Questionnaire for Eating Disorder Diagnoses (Q-EDD). Results of a hierarchical regression analysis revealed PA and AC scores were not statistically significant predictors of ED diagnostic status after controlling for diagnostic subtype. Results of a multiple regression analysis revealed FIDS as a statistically significant predictor of ED diagnostic status when failing to control for ED diagnostic subtype. Discrepancies suggest ED diagnostic subtype may serve as a moderator variable in the relationship between ED diagnostic status and FIDS. PMID- 18425792 TI - Predicting individual differences in autonomy-connectedness: the role of body awareness, alexithymia, and assertiveness. AB - Autonomy-connectedness is the capacity for being on one's own as well as for satisfactorily engaging in interpersonal relationships. Associations have been shown between autonomy-connectedness components (self-awareness, sensitivity to others, and the capacity for managing new situations) and various indices of psychopathology. Both in a theoretical sense as well as for enhancing treatment and prevention, it is relevant to identify which factors most powerfully predict individual differences in autonomy-connectedness: body awareness, alexithymia, or assertiveness. The present study examined this question in a clinical sample of women who were diagnosed as having autonomy problems (N=52) and in a female nonclinical community sample (N=59). In line with expectations, assertiveness was a strong predictor of (all three components of) autonomy-connectedness, as was emotionalizing, one of the alexithymia-components, but the latter in an opposite direction than we had expected: the higher an individual's ability to emotionalize was, the less self-aware and capable to manage new situations that person was, and the more sensitive to others. Cognitive alexithymia contributed to self-awareness as well as to the capacity for managing new situations, and one of the components of body awareness appeared to predict capacity for managing new situations. Our results indicate that assertiveness training and the enhancement of emotion regulation are important elements of autonomy-connectedness targeted interventions. PMID- 18425793 TI - Bcl-2 family proteins: the sentinels of the mitochondrial apoptosis pathway. AB - Bcl-2 family members are the arbiters of mitochondrial apoptotic pathway, which is conserved through evolution. The stoichiometry of pro- versus antiapoptotic Bcl-2 family members in the cell determines whether the cell lives or dies. This fine balance is regulated at the transcriptional or posttranslational level in response to various cellular cues. These signals are transmitted through the upstream molecules in the pathway, that is, the BH3-only molecules that results in the activation of the adaptor molecules, Bax and Bak, at the mitochondrial surface ensuing mitochondrial dysfunction and apoptosis. Understanding the activation process offers a great potential in the therapeutic intervention of many diseases such as cancer and autoimmune disorders. PMID- 18425794 TI - Transcranial Doppler ultrasonography-directed intravenous glycoprotein IIb/IIIa receptor antagonist therapy to control transient cerebral microemboli before and after carotid endarterectomy. AB - BACKGROUND: Patients with a transient focal neurological deficit, critical carotid stenosis and/or microemboli detected by transcranial Doppler ultrasonography (TCD) have a significant risk of stroke. The effect of tirofiban, a selective glycoprotein IIb/IIIa inhibitor, was assessed in patients with microembolic signals on TCD after transient ischaemic attacks or carotid endarterectomy (CEA). METHODS: Thirty-three patients with microemboli on TCD (13 symptomatic preoperative, 19 postoperative, one both) were treated with tirofiban between 2002 and 2007. All patients had carotid stenosis greater than 70 per cent. TCD monitoring was used during and after tirofiban therapy. RESULTS: The median (range) rate of microemboli decreased from 22 (4-260) per h before surgery and 81 (44-216) per h after surgery to 0 (0-9) per h in both groups (P < 0.001, Mann-Whitney U test). This occurred rapidly (preoperative median 30 min; postoperative median 45 min) and was well tolerated in all patients, with no serious adverse effects. CONCLUSION: Cerebral microemboli were controlled by tirofiban both before and after CEA. Further study is required to compare the relative efficacy of tirofiban and dextran. PMID- 18425795 TI - Use of enoxaparin results in more haemorrhagic complications after breast surgery than unfractionated heparin. AB - BACKGROUND: Low molecular weight heparin (LMWH) is used in preference to unfractionated heparin (UFH) for the prevention of postoperative thromboembolism in many UK surgical units. There are, however, conflicting reports on the relative risk of significant bleeding in surgical patients, and no data exist in the literature for patients undergoing breast surgery. METHODS: Data for patients in the Edinburgh Breast Unit with postoperative breast haematoma that needed surgical intervention were analysed for two 12-month intervals in which either UFH (2001) or LMWH (2005-2006) was used for thromboprophylaxis. Haematoma rates in the 6 months after UFH was reintroduced in 2006-2007 were also determined. RESULTS: The rate of haematoma requiring surgical intervention was 0.4 per cent (six of 1452 wounds) in patients who had UFH, compared with 1.8 per cent (32 of 1780 wounds) for LMWH. The rate fell to 0.5 per cent (four of 773 wounds) on reinstituting UFH. The relative risk of haematoma was significantly higher with LMWH than with UFH (4.00 (95 per cent confidence interval 1.97 to 8.11); P < 0.001). No significant postoperative thromboembolic complications were recorded. CONCLUSION: LMWH thromboprophylaxis was associated with a significant increase in haemorrhagic complications after breast surgery compared with UFH. PMID- 18425796 TI - Botulinum toxin for recurrent anal fissure following lateral internal sphincterotomy. AB - BACKGROUND: The aim of the study was to evaluate the efficacy of botulinum toxin injection in the treatment of recurrent anal fissure following lateral internal sphincterotomy. METHODS: Eighty patients were treated with botulinum toxin (30 units Botox or 90 units Dysport), injected into two sites of the internal sphincter. Clinical and manometric results were recorded before and after treatment. If symptoms persisted at 2 months, the examiners could decide to re treat the patient. The same preparation of serotype A of botulinum neurotoxin was used for reinjection. RESULTS: One month after injection there was complete healing in 54 patients (68 per cent). Eight patients (10 per cent) reported mild incontinence of flatus that had disappeared spontaneously within 2 months. At 2 months, 59 patients (74 per cent) had a healing scar. After reinjection, 11 of 21 re-treated patients reported mild incontinence to flatus that lasted for a few weeks and resolved spontaneously. Anorectal manometry at 1 month demonstrated a significant reduction in both resting anal pressure and maximum voluntary squeeze pressure (P < 0.001). There were no relapses during a mean value of 57.9 months of follow-up. CONCLUSION: Botulinum toxin is efficacious in patients with recurrent anal fissure following lateral internal sphincterotomy. PMID- 18425798 TI - Substituent effects on the ring-chain tautomerism of some 1,3-oxazolidine derivatives. AB - The 14 and 70 eV electron ionization mass spectra of five sets (R1 = Me, Et, i Pr, t-Bu and Ph) of seven 2-aryl-4-R1-substituted (Ar = C6H4X; X = p-NO2, m-Br, p Cl, H, p-Me, p-OMe and p-NMe2) (1-5) and of seven 2-aryl-5-phenyl-substituted 1,3 oxazolidines (6; for Ar, see above) were recorded to study their ring-chain equilibria in the gas phase. These equilibria were also studied by 1H NMR spectroscopy in CDCl3 for compounds 5 and 6. A few 2,4- and 2,5-dimethyl-2-aryl derivatives (7, 8: Ar = C6H4X; X = m-Br, H and p-OMe) were studied both in CDCl3 and in the gas phase. The main characteristics of the ring-chain equilibria expressed by the variable SigmaRA% of the ring and of the chain form proved to be a strong dependence on the nature of the substituents on C-2 and C-4. The results in the gas phase are compared with those in CDCl3. PMID- 18425797 TI - Mutations in GDF6 are associated with vertebral segmentation defects in Klippel Feil syndrome. AB - Klippel-Feil syndrome (KFS) is a congenital disorder of spinal segmentation distinguished by the bony fusion of anterior/cervical vertebrae. Scoliosis, mirror movements, otolaryngological, kidney, ocular, cranial, limb, and/or digit anomalies are often associated. Here we report mutations at the GDF6 gene locus in familial and sporadic cases of KFS including the recurrent missense mutation of an extremely conserved residue c.866T>C (p.Leu289Pro) in association with mirror movements and an inversion breakpoint downstream of the gene in association with carpal, tarsal, and vertebral fusions. GDF6 is expressed at the boundaries of the developing carpals, tarsals, and vertebrae and within the adult vertebral disc. GDF6 knockout mice are best distinguished by fusion of carpals and tarsals and GDF6 knockdown in Xenopus results in a high incidence of anterior axial defects consistent with a role for GDF6 in the etiology, diversity, and variability of KFS. PMID- 18425799 TI - Immunophenotyping of lymphocyte subsets in the third eyelid tissue in dogs (Canis familiaris): morphological, microvascular, and secretory aspects of this ocular adnexa. AB - The third eyelid is an important adnexa of the eye. The objective of this study was to evaluate (i) morphological aspects (ii) vascularization, and (iii) the immunophenotype of lymphocyte subsets in the third eyelid of dogs. Flow cytometric analysis revealed the presence of three patterns concerning the immunophenotype of the third eyelid tissue. Dogs without ocular insult or deficient tear production might belong to one of the following immunophenotype patterns: I--the number of T-cells that expressed CD3(+) CD8(+) was higher than the number of cells that expressed CD3(+)CD4(+). II--the number of cells CD3(+)C4(+) was higher than the number of cells CD3(+)CD8(+) and in this case a higher number of cells that expressed CD19 were identified. III--Proximity of values of the cells that expressed CD3(+)CD4(+) and CD3(+)CD8(+). These data might suggest that the number of lymphocyte T cells alone should not be considered a direct indicator of the presence of an immune-based inflammation. Besides, a particular population of T-cells does not indicate a particular inflammatory state. The morphological study of the third eyelid revealed a rather uncommon angioarchitecture. The artery that irrigates the eyelid crosses almost the entire length of this structure to achieve its free border, and only then, ramificates deeply towards an inner segmental level. This spatial microvascular arrangement probably results from an adaptation to the fact that the third eyelid, in the medial cantus of the eye, is inwardly compressed into a tiny space. Efficient vascularization is achieved by allowing the first ramifications of the third eyelid artery to run straight to the top. Accini secretor cells of the third eyelid show a mucin content while tubuloacinar cells are mainly serous. PMID- 18425800 TI - Quantitative binomial distribution analyses of nanoscale like-solute atom clustering and segregation in atom probe tomography data. AB - The applicability of the binomial frequency distribution is outlined for the analysis of the evolution nanoscale atomic clustering of dilute solute in an alloy subject to thermal ageing in 3D atom probe data. The conventional chi(2) statistics and significance testing are demonstrated to be inappropriate for comparison of quantity of solute segregation present in two or more different sized system. Pearson coefficient, mu, is shown to normalize chi(2) with respect to sample size over an order of magnitude. A simple computer simulation is implemented to investigate the binomial analysis and infer meaning in the measured value of mu over a series of systems at different solute concentrations and degree of clustering. The simulations replicate the form of experimental data and demonstrate the effect of detector efficiency to significantly underestimate the measured segregation. The binomial analysis is applied to experimental atom probe data sets and complementary simulations are used to interpret the results. PMID- 18425801 TI - Cr:Forsterite-laser-based fiber-optic nonlinear endoscope with higher efficiencies. AB - We demonstrate a beam-scanning nonlinear light endoscope based on a flexible fiber bundle. Excited with a femtosecond Cr:Forsterite laser, the degradation in multiphoton multiharmonic excitation efficiency due to the pulse-broadening effect is significantly reduced without utilizing any external devices. The system resolution has been characterized to be 5.4 microm in the two-photon fluorescence endoscope, limited by the sampling theory. Finally, several image examples have been given. PMID- 18425802 TI - Probiotic Lactobacillus reuteri suppress proinflammatory cytokines via c-Jun. AB - BACKGROUND: Differential immunoregulatory capabilities of probiotic Lactobacillus were explored in the context of pediatric Crohn's disease. Experimental strategies addressed molecular mechanisms of tumor necrosis factor (TNF) suppression in activated macrophages by transcriptional regulation. METHODS: Secreted factors produced by probiotic Lactobacillus reuteri strains were harvested and tested with human monocytes and macrophages. Quantitative immunoassays and real-time reverse-transcriptase polymerase chain reaction (RT PCR) were used to examine relative quantities of human cytokines and TNF mRNA, respectively, and reporter assays assessed transcriptional regulation of TNF by probiotics. DNA-protein macroarrays interrogated probiotic-mediated effects on transcription factor activation. Finally, enzyme-linked immunosorbent assays (ELISAs) and immunoblots examined the involvement of the specific transcription factor AP-1 and its components. RESULTS: Probiotic L. reuteri strain ATCC PTA 6475 demonstrated the ability to potently suppress human TNF production by lipopolysaccharide-activated monocytes and primary monocyte-derived macrophages from children with Crohn's disease. Quantities of the chemokine MCP-1/CCL2 were also reduced by probiotic L. reuteri strain ATCC PTA 6475 in macrophages of children in remission. Quantitative real-time RT-PCR and luciferase reporter assays showed that transcriptional regulation of human TNF was a primary mechanism of probiotic-mediated immunomodulation. Probiotic L. reuteri suppressed TNF transcription by inhibiting activation of MAP kinase-regulated c-Jun and the transcription factor, AP-1. CONCLUSIONS: Human TNF and MCP-1 suppression by probiotic L. reuteri was strain-dependent, and the activation of c-Jun and AP-1 represent primary targets for probiotic-mediated suppression of TNF transcription. This report emphasizes the clonal nature of immunoprobiosis and delineation of a specific immunomodulatory mechanism for probiotic strain selection in future inflammatory bowel disease-oriented clinical trials. PMID- 18425803 TI - Phenotypical and functional study of ghrelin and its receptor in the pathogenesis of Crohn's disease. AB - BACKGROUND: Ghrelin, a novel endogenous ligand for the growth hormone secretagogue receptor (GHSR), has been demonstrated to possess multiple functions including antiinflammatory effects. The aim of this study was to investigate the expression of ghrelin and GHSR and the function of ghrelin in inflammatory bowel disease (IBD). METHODS: The expression of ghrelin and GHSR mRNA was quantified in mucosal biopsy specimens from 9 controls, 15 patients with Crohn's disease (CD), and 15 patients with ulcerative colitis (UC) using quantitative reverse transcriptase polymerase chain reaction (RT-PCR). The locations of ghrelin and GHSR were investigated immunohistochemically in surgically resected specimens. We also evaluated the percentage of GHSR-positive peripheral blood mononuclear cells (PBMCs) in healthy controls and patients with CD by flow cytometry. In addition, we investigated the immunoregulatory function of ghrelin in peripheral blood T cells. RESULTS: Ghrelin mRNA levels in colonic mucosa of IBD were higher than control level. The GHSR-1a mRNA level in active CD was also significantly higher than the control level. Ghrelin and GHSR-1a were expressed on CD3- and CD68 positive cells. The percentage of GHSR-1a-positive peripheral blood T cells in patients with CD was significantly higher than the control level. Stimulation of human T cells with ghrelin increased levels of IL-4 and IL-13 proteins and decreased levels of IFN-gamma protein. Reactivity to ghrelin was low in CD compared with the control level. CONCLUSIONS: Our findings demonstrate that ghrelin may play an important role in the immune system in CD. The dysregulation of reactivity of T cells induced by ghrelin suggests that ghrelin might participate in the pathogenesis of CD. PMID- 18425805 TI - Novel model for studying hematogenous infection in an experimental setting of implant-related infection by a community-acquired methicillin-resistant S. aureus strain. AB - The aim of this study is to establish a new experimental model of hematogenous implant-related infection (IRI) by a community-acquired methicillin-resistant S. aureus (CA-MRSA) strain. Cylindrical porous tantalum intramedullary implants were inserted in the proximal right tibia of 30 male white rabbits after administration of antibiotic prophylaxis. Four weeks later and without antibiotic prophylaxis, 20 animals received 1 ml of inoculum of two different concentrations (study groups A and B) of CA-MRSA strain through an ipsilatelar femoral artery catheter. The remaining 10 received normal saline instead (control group C). Surviving animals were sacrificed 4 weeks later. Sterile bone, bone marrow biopsies, and implants were harvested for culture and histological evaluation. Ten animals receiving 5 x 10(8)cfu/ml (group A) died within 48-72 h due to septic shock. Blood cultures were positive; histology demonstrated acute infection. Ten animals received bacterial load of 3 x 10(8)cfu/ml (group B) and all survived; two had negative Gram-stain and cultures but PCR and RT-PCR results demonstrated the viability of the microorganisms, while periprosthetic osteolysis and histological evaluation indicated subacute osteomyelitis; eight animals established periprosthetic infection, osteomyelitis, and septic arthritis documented by positive Gram-stain, cultures, subperiosteal reaction, and chronic infection on histology. Control group specimens demonstrated no signs of infection. Histopathological semiquantitative scoring was used to compare the three groups. Comparison of groups A and B with control group and between group A and B showed statistically significant difference (p < 0.05) in all parameters except for periosteal reaction between groups B and C (p = 0.354). This novel, reproducible experimental model will facilitate the study of hematogenous CA-MRSA IRIs. PMID- 18425804 TI - Glycine receptor-mediated synaptic transmission regulates the maturation of ganglion cell synaptic connectivity. AB - It is well documented that neuronal activity is required for the developmental segregation of retinal ganglion cell (RGC) synaptic connectivity with ON and OFF bipolar cells in mammalian retina. Our recent study showed that light deprivation preferentially blocked the developmental RGC dendritic redistribution from the center to sublamina a of the inner plexiform layer (IPL). To determine whether OFF signals in visual stimulation are required for OFF RGC dendritic development, the light-evoked responses and dendritic stratification patterns of RGCs in Spastic mutant mice, in which the OFF signal transmission in the rod pathway is largely blocked due to a reduction of glycine receptor (GlyR) expression, were quantitatively studied at different ages and rearing conditions. The dendritic distribution in the IPL of these mice was indistinguishable from wildtype controls at the age of postnatal day (P)12. However, the adult Spastic mutants had altered RGC light-evoked synaptic inputs from ON and OFF pathways, which could not be mimicked by pharmacologically blocking of glycinergic synaptic transmission on age-matched wildtype animals. Spastic mutation also blocked the developmental redistribution of RGC dendrites from the center to sublamina a of the IPL, which mimicked the effects induced by light deprivation on wildtype animals. Moreover, light deprivation of the Spastic mutants had no additional impact on the RGC dendritic distribution and light response patterns. We interpret these results as that visual stimulation regulates the maturation of RGC synaptic activity and connectivity primarily through GlyR-mediated synaptic transmission. PMID- 18425806 TI - Common excipients impair detection of protein aggregates during sedimentation velocity analytical ultracentrifugation. AB - The final formulations of modern pharmaceutical protein products typically contain sugars or sugar alcohols as stabilizers. Migration of these sugars under the influence of an applied gravitational field during sedimentation velocity analytical ultracentrifugation (SV-AUC) produces dynamic density and viscosity gradients. If the formation of such gradients is not taken into account during data analysis, the capability of the SV-AUC technique to detect protein oligomers/aggregates may be dramatically impacted. In the example described here, the limit of quantitation (LOQ) of a simulated monoclonal antibody (mAb) dimer increases from 0.8% to 2.4% upon addition of 5% sorbitol to the formulation. This study uses simulated and experimental SV-AUC data to demonstrate the detrimental effect of dynamic gradients; it further explores how sophisticated data analysis techniques, including SEDFIT's inhomogeneous solvent options, may be used to mitigate the detection problems caused by the sedimentation of excipients. PMID- 18425807 TI - Controlled release of repifermin from polyelectrolyte complexes stimulates endothelial cell proliferation. AB - The therapeutic value of many growth factors is often hindered by the narrow therapeutic index and sustained concentrations required for efficacy. Controlled release approaches provide a valuable tool to achieve these goals; however, growth factor stability must be maintained. Repifermin is a truncated form of fibroblast growth factor-10, also known as keratinocyte growth factor-2, that exhibits promise in wound healing applications; however, controlled release formulation presents a challenge for this labile protein. Taking advantage of the heparin-binding motif of this class of biopharmaceuticals, Repifermin was effectively stabilized and packaged in polyelectrolyte complexes. In the presence of dextran sulfate, the unfolding temperature of this growth factor was increased by approximately 10 degrees C as confirmed by a variety of spectroscopic techniques. Dextran sulfate with bound Repifermin was then complexed with several polycations (chitosan, poly-L-lysine, and polyethylenimine) resulting in the formation of approximately 250 nm polyelectrolyte complexes that entrapped the protein with approximately 70-80% efficiency. Release was controlled for more than 10 days and the mitogenic activity of Repifermin on human umbilical cord vascular endothelial cells was significantly enhanced, whereas no effect was noted for free Repifermin. PMID- 18425808 TI - Complete protection of sodium valproate from humidity by using a hydrotalcite composite. AB - Complete protection of sodium valproate from humidity was successfully accomplished by intercalation into a hydrotalcite-like compound. The obtained hybrid of sodium valproate and the hydrotalcite-like compound was stable and not at all hygroscopic. Even under a relative humidity of 97%, the adsorbed amount of water was less than 0.5%. PMID- 18425809 TI - Structural remodeling of nucleus ambiguus projections to cardiac ganglia following chronic intermittent hypoxia in C57BL/6J mice. AB - The baroreflex control of heart rate (HR) is reduced following chronic intermittent hypoxia (CIH). Since the nucleus ambiguus (NA) plays a key role in baroreflex control of HR, we examined whether CIH remodels vagal efferent projections to cardiac ganglia. C57BL/6J mice (3-4 months of age) were exposed to either room air (RA) or CIH for 3 months. Confocal microscopy was used to examine NA axons and terminals in cardiac ganglia following Fluoro-Gold (FG) injections to label cardiac ganglia, and microinjections of tracer DiI into the left NA to anterogradely label vagal efferents. We found that: 1) Cardiac ganglia were widely distributed on the dorsal surface of the atria. Although the total number of cardiac ganglia did not differ between RA and CIH mice, the size of ganglia and the somatic area of cardiac principal neurons (PNs) were significantly decreased (P < 0.01), and the size of the PN nuclei was increased following CIH (P < 0.01). 2) NA axons entered cardiac ganglia and innervated PNs with dense basket endings in both RA and CIH mice, and the percentage of innervated PNs was similar (RA: 50 +/- 1.0%; CIH: 49 +/- 1.0%; P > 0.10). In CIH mice, however, swollen cardiac axons and terminals without close contacts to PNs were found. Furthermore, varicose endings around PNs appeared swollen and the axonal varicose area around PNs was almost doubled in size (CIH: 163.1 +/- 6.4 microm(2); RA: 88 +/- 3.9 microm(2), P < 0.01). Thus, CIH significantly altered the structure of cardiac ganglia and resulted in reorganized vagal efferent projections to cardiac ganglia. Such remodeling of cardiac ganglia and vagal efferent projections provides new insight into the effects of CIH on the brain-heart circuitry of C57BL/6J mice. PMID- 18425810 TI - Peer victimization and depression among Hong Kong adolescents. AB - The present study examined the gender differences in the relationship between victimization and depression, as well as in the buffering effect of social support, in Hong Kong adolescents. Based on social role theory, we predicted (a) that victimization would be associated with higher depression in boys than in girls, and (b) that social support would have a stronger buffering effect in boys than in girls. Results showed that boys reported more victimizations than girls, and victimization was slightly more strongly associated with depressive symptoms in boys than in girls. Parental support had a direct effect on depression, but no buffering effect. A buffering effect was found for friend support, but only in boys; high support from friends was associated with fewer depressive symptoms at high levels of victimization. PMID- 18425811 TI - Immunolocalization of NMDA receptor subunit NR3B in selected structures in the rat forebrain, cerebellum, and lumbar spinal cord. AB - N-methyl-D-aspartate (NMDA) receptors have been implicated in many neurological disorders. Although NMDA receptors are best known for their high calcium permeability, the recently discovered NR3 subunits, NR3A and NR3B, have been shown to reduce the calcium permeability of the NMDA receptor. Thus, NR3 subunits may be important players in modulating synaptic plasticity in neurons. Although NR3B expression in the rodent and human brain has been studied, little is known about its distribution in different cell types. Here we used immunolabeling with a specific NR3B antibody together with antibodies against established neurochemical markers to determine the cellular and subcellular localization of NR3B. The nucleus was concurrently stained with NR3B immunolabeling to show that NR3B is widely expressed by many cells in each brain region. Our findings indicate that NR3B is widely expressed in the structures examined in the rat forebrain (hippocampus, cerebral cortex, caudoputamen, and nucleus accumbens), cerebellum, and lumbar sections of the spinal cord. Within these regions NR3B was found to be expressed in all the substructures of the hippocampus (CA1, CA3, dentate gyrus), the various layers of the cerebral cortex, projection neurons and interneurons of the striatum, different cell types of the cerebellum, and motor neurons of the spinal cord. Furthermore, when stained with NR1-the obligatory subunit responsible for forming functional NMDA receptors-the distribution of NR3B appears to be as ubiquitous as NR1. Taken together, our data suggest that there may be a population of NR3B-containing NMDA receptors conferring new functional roles in the mammalian central nervous system. PMID- 18425813 TI - Zidovudine-loaded PLA and PLA-PEG blend nanoparticles: influence of polymer type on phagocytic uptake by polymorphonuclear cells. AB - Mononuclear (macrophages) and polymorphonuclear leucocytes cells play an important role in the immunopathogenesis of acquired immunodeficiency syndrome. Zidovudine is a broad-spectrum drug used in current antiretroviral therapy. The development of controlled drug delivery systems for the treatment of chronic diseases is of great interest since these systems can act as vectors, carrying the drug only to the target, and the adverse effects can be reduced. In this study, PLA and PLA/PEG blend nanoparticles containing zidovudine were developed and their uptake by polymorphonuclear leucocytes were studied in vitro. The influence of polymer type on particle size, Zeta potential and particle uptake by polymorphonuclear leucocytes was investigated. The cells were isolated from rat peritoneal exudate and their activation by nanoparticles was measured by luminol dependent chemiluminescence and microscopical analysis. The PEG in the blend modified the Zeta potential suggested the formation of a PEG coat on the particle surface. The phagocytosis depended on the PEG and its ratio in the blend, the results showed that the PLA nanoparticles were more efficiently phagocytosed than PLA/PEG blends. The blend with the highest PEG proportion did not prevent phagocytosis, indicating that the steric effect of PEG was concentration dependent. PMID- 18425814 TI - Biowaiver monographs for immediate release solid oral dosage forms: aciclovir. AB - Literature data relevant to the decision to allow a waiver of in vivo bioequivalence (BE) testing (biowaiver) for the approval of immediate release (IR) solid oral dosage forms containing aciclovir are reviewed. Aciclovir therapeutic use and therapeutic index, pharmacokinetic properties, data related to the possibility of excipient interactions and reported BE/bioavailability (BA) studies were also taken into consideration in order to ascertain whether a biowaiver can be recommended. According to the Biopharmaceutics Classification System (BCS) and considering tablet strengths up to 400 mg, aciclovir would be BCS Class III. However, in some countries also 800 mg tablets are available which fall just within BCS Class IV. Aciclovir seems not to be critical with respect to a risk for bioinequivalence, as no examples of bioinequivalence have been identified. It has a wide therapeutic index and is not used for critical indications. Hence, if: (a) the test product contains only excipients present in aciclovir solid oral IR drug products approved in ICH or associated countries, for instance as presented in this article; and (b) the comparator and the test product both are very rapidly dissolving, a biowaiver for IR aciclovir solid oral drug products is considered justified for all tablet strengths. PMID- 18425815 TI - PLGA nanospheres for the ocular delivery of flurbiprofen: drug release and interactions. AB - Poly(D,L-lactide-co-glycolide) nanospheres incorporating flurbiprofen were prepared by the solvent displacement technique for purposes of assessing (i) drug polymer physicochemical interactions, (ii) flurbiprofen release from the polymer matrix and (iii) eye permeation of the drug formulated in the colloidal system. The resulting nanospheres were on average 200-300 nm in size and bore a negative charge (xi-potential around -25 mV). They were shown by atomic force microscopy and transmission electron microscopy to be spherical and regular in shape. Thermal methods, infrared spectroscopy and X-ray diffraction showed that the drug was dispersed inside the particles. These tests evidenced an eutectic mixture meaning more widespread dispersion of the drug in the polymer system. Entrapped flurbiprofen was released in vitro from the polymer system by dissolution and diffusion in high drug loaded nanospheres, whereas those with a lesser load showed only diffusion. The ex vivo corneal permeation study showed that flurbiprofen-loaded nanospheres enhanced drug penetration by about twofold over commercial eye drops containing poly(vinyl alcohol) and by about fourfold over flurbiprofen in pH 7.4 phosphate buffer. The corneal hydration level of each cornea was determined to evaluate potential corneal damage. PMID- 18425817 TI - The aberrant methylation of TSP1 suppresses TGF-beta1 activation in colorectal cancer. AB - Colorectal cancer arises from the progressive accumulation of mutations and epigenetic alterations in colon epithelial cells. Such alterations often deregulate signaling pathways that affect the formation of colon cancer, such as the Wnt, RAS-MAPK and TGF-beta pathways. The tumor promoting effects of mutations in genes, such as APC, have been demonstrated in cancer cell lines and in mouse models of intestinal cancer; however, the biological effects of most epigenetic events identified in colorectal cancer remain unknown. Consequently, we assessed whether the aberrant methylation of TSP1, the gene for thrombospondin 1, a regulator of TGF-beta ligand activation, is an epigenetic mechanism for inhibiting the TGF-beta signaling pathway. We found methylated TSP1 occurs in colon cancer cell lines (33%), colon adenomas (14%) and colon adenocarcinomas (21%). In primary colorectal cancers, loss of TSP1 expression correlated with impaired TGF-beta signaling as indicated by decreased Smad2 phosphorylation and nuclear localization. Furthermore, methylation-induced silencing of TSP1 expression reduced the concentration of secreted active TGF-beta1 and attenuated TGF-beta signaling. Reversal of TSP1 methylation resulted in increased TSP1 mediated activation of the latent LAP:TGF-beta complex and subsequent TGF-beta receptor activation. Our results demonstrate that the aberrant methylation of TSP1 has biological consequences and provide evidence that the aberrant methylation of TSP1 is a novel epigenetic mechanism for suppressing TGF-beta signaling in colorectal cancer. PMID- 18425818 TI - Modes of action of the DNA methyltransferase inhibitors azacytidine and decitabine. AB - The cytosine analogues 5-azacytosine (azacytidine) and 2'-deoxy-5-azacytidine (decitabine) are the currently most advanced drugs for epigenetic cancer therapies. These compounds function as DNA methyltransferase inhibitors and have shown substantial potency in reactivating epigenetically silenced tumor suppressor genes in vitro. However, it has been difficult to define the mode of action of these drugs in patients and it appears that clinical responses are influenced both by epigenetic alterations and by apoptosis induction. To maximize the clinical efficacy of azacytidine and decitabine it will be important to understand the molecular changes induced by these drugs. In this review, we examine the pharmacological properties of azanucleosides and their interactions with various cellular pathways. Because azacytidine and decitabine are prodrugs, an understanding of the cellular mechanisms mediating transmembrane transport and metabolic activation will be critically important for optimizing patient responses. We also discuss the mechanism of DNA methyltransferase inhibition and emphasize the need for the identification of predictive biomarkers for the further advancement of epigenetic therapies. PMID- 18425819 TI - Lung cancer epigenetics and genetics. AB - Lung cancer is the leading cause of cancer-related death and thus a major health problem. The efficiency of current treatment modalities for lung cancer depends strongly on the time of diagnosis, with better chances of survival if a tumor has been detected at an early stage. Thus, there is an urgent need for rapid and efficient early detection methods. Biomarkers represent a possible alternative to current, rather expensive, screening tools such as spiral computer tomography (CT), or may allow the identification of high risk groups for whom screening would be cost efficient. Although most lung cancers are the consequence of smoking, a substantial fraction of molecular-epidemiological studies point to high-prevalence, low-penetrance genetic polymorphisms as modifiers of environmental lung cancer risk. In the past the genomics field has also made significant advances in identifying genetic lesions that can now be harvested with the goal of identifying novel biomarkers for lung cancer. Furthermore, the importance of epigenetic changes that occur during lung cancer development has been reported, but has been underestimated in the past. Novel high-throughput, quantitative assays for the detection of DNA methylation or histone tail modifications are now applied, to search for alterations in the lung cancer genome and will identify novel cancer-related genes that may become attractive targets for treatment, provide new insight into the biology of lung cancers, and could also become useful biomarkers for the early detection of lung cancer in sputum, or may be used as prognostic markers. Thus, an integrative approach in lung cancer research combining epidemiological, genetic and epigenetic information becomes an important concept for the future. PMID- 18425820 TI - Isotype-specific inhibitors of the glycolytic key regulator pyruvate kinase subtype M2 moderately decelerate tumor cell proliferation. AB - Tumor cells express the glycolytic regulator pyruvate kinase subtype M2 (M2-PK), which can occur in a tetrameric form with high affinity to its substrate phosphoenolpyruvate (PEP) and a dimeric form with a low PEP affinity. The transition between both conformations contributes to the control of glycolysis and is important for tumor cell proliferation and survival. Here we targeted M2 PK by synthetic peptide aptamers, which specifically bind to M2-PK and shift the isoenzyme into its low affinity dimeric conformation. The aptamer-induced dimerization and inactivation of M2-PK led to a significant decrease in the PK mass-action ratio as well as ATP:ADP ratio in the target cells. Furthermore, the expression of M2-PK-binding peptide aptamers moderately reduced the growth of immortalized NIH3T3 cell populations by decelerating cell proliferation, but without affecting apoptotic cell death. Moreover, the M2-PK-binding peptide aptamers also reduced the proliferation rate of human U-2 OS osteosarcoma cells. In the present study, we developed the first specific inhibitors of the pyruvate kinase isoenzyme type M2 and present evidence that these inhibitors moderately decelerate tumor cell proliferation. PMID- 18425821 TI - On multiple-testing correction in genome-wide association studies. AB - The interpretation of the results of large association studies encompassing much or all of the human genome faces the fundamental statistical problem that a correspondingly large number of single nucleotide polymorphisms markers will be spuriously flagged as significant. A common method of dealing with these false positives is to raise the significance level for the individual tests for association of each marker. Any such adjustment for multiple testing is ultimately based on a more or less precise estimate for the actual overall type I error probability. We estimate this probability for association tests for correlated markers and show that it depends in a nonlinear way on the significance level for the individual tests. This dependence of the effective number of tests is not taken into account by existing multiple-testing corrections, leading to widely overestimated results. We demonstrate a simple correction for multiple testing, which can easily be calculated from the pairwise correlation and gives far more realistic estimates for the effective number of tests than previous formulae. The calculation is considerably faster than with other methods and hence applicable on a genome-wide scale. The efficacy of our method is shown on a constructed example with highly correlated markers as well as on real data sets, including a full genome scan where a conservative estimate only 8% above the permutation estimate is obtained in about 1% of computation time. As the calculation is based on pairwise correlations between markers, it can be performed at the stage of study design using public databases. PMID- 18425822 TI - MRI abnormalities in normal-appearing brain tissue of treated adult PKU patients. AB - PURPOSE: To detect possible subclinical pathological brain changes a study on adult phenylketonuria (PKU) patients by using quantitative MRI methods was performed, since neuropsychological and cognitive deficits in treated patients with PKU have not yet been shown to correlate clearly with the brain lesions identified by conventional MRI. MATERIALS AND METHODS: Eight subjects, four PKU patients with well-documented dietary treatment and four age- and sex-matched adult controls, underwent MRI, including a triple echo sequence and a diffusion tensor imaging sequence. Brain maps of T2 relaxation time (T2), relative proton density (PD), and fractional anisotropy (FA) as well as apparent diffusion coefficient (ADC) were derived for each subject. T2, PD, FA, and ADC were measured in 22 predefined regions of gray matter (GM) and white matter (WM) on the corresponding maps, and compared with those of four age-matched healthy adult controls. RESULTS: In addition to a prolonged T2 value measured in affected WM, as expected, we observed a significant shortening of the T2 relaxation time and reduction of ADC in normal-appearing brain tissue and an increased proton density in both GM and WM of the patients. No differences were observed in FA values between controls and patients. CONCLUSION: Repeatedly reduced T2 relaxation time, ADC, and increased proton density without changes in FA indicate a higher cell packing density in normal-appearing brain without changes in the directedness of fibers. These structural changes may be related to neuropsychological and cognitive deficits in treated PKU patients. PMID- 18425823 TI - 3D myocardial tissue tracking with slice followed cine DENSE MRI. AB - PURPOSE: To track three-dimensional (3D) myocardial tissue motion using slice followed cine displacement encoded imaging with stimulated echoes (DENSE). MATERIALS AND METHODS: Slice following (SF) has previously been developed for 2D myocardial tagging to compensate for the effect of through-plane motion on 2D tissue tracking. By incorporating SF into a cine DENSE sequence, and applying displacement encoding in three orthogonal directions, we demonstrate the ability to track discrete elements of a slice of myocardium in 3D as the heart moves through the cardiac cycle. The SF cine DENSE tracking algorithm was validated on a moving phantom, and the effects of through-plane motion on 2D cardiac strain were investigated in six healthy subjects. RESULTS: A through-plane tracking accuracy of 0.46 +/- 0.32 mm was measured for a typical range of myocardial motion using a rotating phantom. In vivo 3D measurements of cardiac motion were consistent with prior myocardial tagging results. Through-plane rotation in a mid ventricularshort-axis view was shown to decrease the magnitude of the 2D end systolic circumferential strain by 3.91 +/- 0.43% and increase the corresponding radial strain by 6.01 +/- 1.07%. CONCLUSION: Slice followed cine DENSE provides an accurate method for 3D tissue tracking. PMID- 18425824 TI - Adaptations in trabecular bone microarchitecture in Olympic athletes determined by 7T MRI. AB - PURPOSE: To produce in vivo high-resolution images of the knee and to determine the feasibility of using 7T MR to detect changes in trabecular bone microarchitecture in elite athletes (Olympic fencers) who undergo high impact activity. MATERIALS AND METHODS: The dominant knees of four males from the U.S. Olympic Fencing Team and three matched healthy male controls were scanned in a 7T whole-body scanner using a quadrature knee coil with three-dimensional (3D) fast low angle shot (FLASH): 50 axial images at the distal femur (0.156 mm x 0.156 mm) and 80 axial images at the knee joint (0.195 mm x 0.195 mm). Bone volume fraction (BVF) and marrow volume fraction (MVF) images were computed and fuzzy distance transform (FDT) and digital topological analysis (DTA) were applied to determine: trabecular number (Tb.N), trabecular thickness (Tb.Th), and trabecular separation (Tb.Sp); BVF (BV/TV); trabecular and marrow space surface-to-curve ratio (SC, marker of plate to rod ratio); and trabecular and marrow space erosion index (EI, inverse marker for network connectivity). Quadriceps muscle volume (MV) was calculated as well. We calculated group means and performed two-tailed t-tests to determine statistical significance. RESULTS: Compared to controls, fencers had: decreased Tb.Sp (P = 0.0082 at femur, P = 0.051 at joint); increased Tb.N (P < 0.05 at both femur and joint) and BV/TV (P < 0.001 at both femur and joint); increased trabecular SC and decreased marrow space SC (P < 0.01 at both femur and joint); decreased trabecular EI and increased marrow space EI (P < 0.01 at both femur and joint); and increased MV (P = 0.038). There was no difference in Tb.Th at the distal femur (P = 0.92) or joint (P = 0.71) between groups. CONCLUSION: To our knowledge, this is the first study to perform 7T MRI of the knee in vivo. Elite athletes who undergo high impact activity have increased MV and improved trabecular bone structure compared to controls. PMID- 18425825 TI - Effect of vascular stenosis on perfusion-weighted imaging; differences between calculation algorithms. AB - PURPOSE: To determine the most suitable postprocessing technique for magnetic resonance (MR) perfusion imaging in patients with vascular stenosis, by comparing the cerebral blood flow (CBF) maps of single photon emission tomography (SPECT) and perfusion MR imaging (MRI). MATERIALS AND METHODS: In 15 consecutive patients (14 men and one woman, mean age 73.9 +/- 6.0 years) with stenosis of common carotid artery (CCA) or internal carotid artery (ICA) of more than 75%, both brain perfusion MRI and brain perfusion SPECT were performed. From perfusion MR images, CBF maps were calculated with the first moment, singular value decomposition (SVD), and block circulant SVD (b-SVD) methods, and CBF maps from each algorithm were compared with those from SPECT. RESULTS: The b-SVD method had the best correlation with SPECT (R = 0.814), followed by the first moment method (R = 0.776) and the SVD method (R = 0.723). The b-SVD method has the least mean difference with SPECT (0.118), the first moment method also had less difference (0.121), and the SVD had greatest mean difference (0.164). CONCLUSION: Our results suggest that in patients with vascular impairment the b-SVD method will be the technique of choice rather than SVD or first moment method. PMID- 18425826 TI - Water excitation MPRAGE: an alternative sequence for postcontrast imaging of the abdomen in noncooperative patients at 1.5 Tesla and 3.0 Tesla MRI. AB - PURPOSE: To evaluate the diagnostic image quality of postgadolinium water excitation-magnetization-prepared rapid gradient-echo (WE-MPRAGE) sequence in abdominal examinations of noncooperative patients at 1.5 Tesla (T) and 3.0T MRI. MATERIALS AND METHODS: Eighty-nine consecutive patients (48 males and 41 females; mean age +/- standard deviation, 54.6 +/- 16.6 years) who had MRI examinations including postgadolinium WE-MPRAGE were included in the study. Of 89 patients, 33 underwent noncooperative protocol at 1.5T, 10 underwent noncooperative protocol at 3.0T, and 46 underwent cooperative protocol at 3.0T. Postgadolinium WE-MPRAGE, MPRAGE, and three-dimensional gradient-echo sequences of these three different groups were qualitatively evaluated for image quality, extent of artifacts, lesion conspicuity, and homogeneity of fat-attenuation by two reviewers retrospectively, independently, and blindly. The results were compared using Wilcoxon signed rank and Mann-Whitney U tests. Kappa statistics were used to measure the extent of agreement between the reviewers. RESULTS: The average scores indicated that the images were diagnostic for WE-MPRAGE at 1.5T and 3.0T in noncooperative patients. WE-MPRAGE achieved homogenous fat-attenuation in 31/33 (94%) of noncooperative patients at 1.5T and 10/10 (100%) of noncooperative patients at 3.0T. WE-MPRAGE at 3.0T had better results for image quality, extent of artifacts, lesion conspicuity and homogeneity of fat-attenuation compared with WE-MPRAGE at 1.5T, in noncooperative patients (P = 0.0008, 0.0006, 0.0024, and 0.0042; respectively). Kappa statistics varied between 0.76 and 1.00, representing good to excellent agreement. CONCLUSION: WE-MPRAGE may be used as a T1-weighted postgadolinium fat-attenuated sequence in noncooperative patients, particularly at 3.0T MRI. PMID- 18425827 TI - Positive contrast MR-lymphography using inversion recovery with ON-resonant water suppression (IRON). AB - PURPOSE: To investigate the utility of inversion recovery with ON-resonant water suppression (IRON) to create positive signal in normal lymph nodes after injection of superparamagnetic nanoparticles. MATERIALS AND METHODS: Experiments were conducted on six rabbits, which received a single bolus injection of 80 mumol Fe/kg monocrystalline iron oxide nanoparticle (MION-47). Magnetic resonance imaging (MRI) was performed at baseline, 1 day, and 3 days after MION-47 injection using conventional T(1)- and T(2)*-weighted sequences and IRON. Contrast-to-noise ratios (CNR) were measured in blood and in paraaortic lymph nodes. RESULTS: On T(2)*-weighted images, as expected, signal attenuation was observed in areas of paraaortic lymph nodes after MION-47 injection. However, using IRON the paraaortic lymph nodes exhibited very high contrast enhancement, which remained 3 days after injection. CNR with IRON was 2.2 +/- 0.8 at baseline, increased markedly 1 day after injection (23.5 +/- 5.4, P < 0.01 vs. baseline), and remained high after 3 days (21.8 +/- 5.7, *P < 0.01 vs. baseline). CNR was also high in blood 1 day after injection (42.7 +/- 7.2 vs. 1.8 +/- 0.7 at baseline, P < 0.01) but approached baseline after 3 days (1.9 +/- 1.4, P = NS vs. baseline). CONCLUSION: IRON in conjunction with superparamagnetic nanoparticles can be used to perform 'positive contrast' MR-lymphography, particularly 3 days after injection of the contrast agent, when signal is no longer visible within blood vessels. The proposed method may have potential as an adjunct for nodal staging in cancer screening. PMID- 18425828 TI - Single-breathhold four-dimensional assessment of left ventricular volumes and function using k-t BLAST after application of extracellular contrast agent at 3 Tesla. AB - PURPOSE: To prospectively determine the feasibility and accuracy of a four dimensional (4D) k-space over time broad-use linear acquisition speed-up technique (k-t BLAST) for the evaluation of left ventricular (LV) volumes in comparison to standard multiple-breathhold cine imaging, using a 3.0 Tesla (3T) MR system. MATERIALS AND METHODS: In 23 subjects, short-axis cine loops completely covering the LV were acquired using conventional turbo gradient echo (GRE) imaging. Immediately after administration of gadobenate dimeglumine, a rapid single-breathhold k-t BLAST 4D dataset with the same coverage was acquired and reconstructed to short-axis views. Quantitative aortic flow measurement for LV stroke volume (LVSV) was used to calibrate both techniques. For GRE and k-t BLAST cine imaging: LV volumes, ejection fraction (EF), and blood-to-myocardium contrast (BMC) were determined. RESULTS: k-t BLAST and GRE sequences showed a strong correlation for LV volumes and EF (r = 0.97-0.99; P < 0.001). Excellent agreement was also found between the LVSV determined by aortic flow measurements and LVSV assessed using GRE sequence and k-t BLAST sequence. BMC of GRE was similar to that of k-t BLAST cine imaging. CONCLUSION: The use of the single breathhold 4D k-t BLAST technique for the assessment of LV volume is feasible and accurate in 3T MRI. PMID- 18425829 TI - Determining exercise-induced blood flow reserve in lower extremities using phase contrast MRI. AB - PURPOSE: To study the changes in limb blood flow during lower extremity exercise using phase contrast (PC) MRI in normal volunteers. MATERIALS AND METHODS: Healthy volunteers performed plantar flexion exercise (<1 W) for four minutes. Flow velocity was measured using cardiac-gated, cine PC-MRI sequences (fast gradient recalled echo [GRE]; multishot echo planar imaging [EPI]) on a 3T scanner at the level of the superficial femoral artery (SFA): 1) preexercise; 2) immediately postexercise; 3) during three minutes recovery; and 4) postrecovery. RESULTS: At rest there was a triphasic flow waveform in the SFA. During exercise it changed to a monophasic pattern with an increase in total flow; there were variable changes in vessel size and flow velocity. The waveform regained the triphasic pattern during recovery. The exercise-induced flow reserve (FR) was 167 +/- 90%. CONCLUSION: PC-MRI demonstrates that the resting triphasic flow waveform transforms into a monophasic pattern with submaximal exercise and returns to baseline with recovery. This increase in the regional blood flow allows for measurement of exercise-induced FR in the SFA. PMID- 18425830 TI - Interstudy reproducibility of SSFP cine magnetic resonance: impact of magnetic field strength and parallel imaging. AB - PURPOSE: To study the effects of field strength and parallel imaging on image contrast and interstudy reproducibility of right and left ventricular (RV and LV) measurements using steady-state free precession (SSFP) cardiovascular magnetic resonance (CMR). MATERIALS AND METHODS: Thirty-two subjects (20 normal, 12 cardiac patients) underwent four SSFP cine short-axis imaging studies: two at 1.5T, one at 3T, and another at 3T with parallel imaging (SENSE). RV and LV contrast-to-noise ratios (CNRs) were compared between methods. Interstudy reproducibility of RV and LV measurements were assessed by Bland-Altman analyses. RESULTS: 3T SENSE imaging reduced total imaging time from 8 +/- 2 minutes to 3 +/ 1 minute (P < 0.001). A significant gain in LV CNR was detected between 1.5T and 3T with SENSE (43.8 +/- 6.5 vs. 48.4 +/- 7.4, P = 0.01), but no significant gain was detected in RV CNR. The reproducibility of LV and RV measurements between two 1.5T studies was not significantly different from the reproducibility between a 1.5T study and a 3T study with SENSE. CONCLUSION: SENSE imaging at 3T is a reproducible technique for assessing RV and LV structure and function. PMID- 18425831 TI - Correction for heart rate variability during 3D whole heart MR coronary angiography. AB - PURPOSE: To evaluate the effect of a real-time adaptive trigger delay on image quality to correct for heart rate variability in 3D whole-heart coronary MR angiography (MRA). MATERIALS AND METHODS: Twelve healthy adults underwent 3D whole-heart coronary MRA with and without the use of an adaptive trigger delay. The moment of minimal coronary artery motion was visually determined on a high temporal resolution MRI. Throughout the scan performed without adaptive trigger delay, trigger delay was kept constant, whereas during the scan performed with adaptive trigger delay, trigger delay was continuously updated after each RR interval using physiological modeling. Signal-to-noise, contrast-to-noise, vessel length, vessel sharpness, and subjective image quality were compared in a blinded manner. RESULTS: Vessel sharpness improved significantly for the middle segment of the right coronary artery (RCA) with the use of the adaptive trigger delay (52.3 +/- 7.1% versus 48.9 +/- 7.9%, P = 0.026). Subjective image quality was significantly better in the middle segments of the RCA and left anterior descending artery (LAD) when the scan was performed with adaptive trigger delay compared to constant trigger delay. CONCLUSION: Our results demonstrate that the use of an adaptive trigger delay to correct for heart rate variability improves image quality mainly in the middle segments of the RCA and LAD. PMID- 18425832 TI - Diffusion-weighted magnetic resonance imaging allows monitoring of anticancer treatment effects in patients with soft-tissue sarcomas. AB - PURPOSE: To determine if diffusion-weighted imaging (DWI) can be used as a surrogate marker of tumor response to anticancer therapy in patients with soft tissue sarcomas. MATERIALS AND METHODS: Magnetic resonance imaging (MRI) including echo-planar DWI sequences was performed prospectively in 23 consecutive patients with soft-tissue sarcomas before and after initiation of regional or systemic chemotherapy. The mean interval between initial and follow-up MRI was 56.9 +/- 23.2 days. Tumor volumes were determined by manual segmentation of tumor borders on contrast-enhanced T1-weighted images. The apparent diffusion coefficient (ADC) was calculated from corresponding sections of ADC maps on initial and follow-up DWI. Subsequently, changes in tumor volumes and ADC were correlated using the Pearson correlation coefficient. RESULTS: A high degree of correlation was found when changes in tumor volumes and ADC values were compared (r = -0.925, P < 0.0001), regardless of the effectiveness of anticancer therapy expressed as changes of tumor volume. CONCLUSION: DWI can be used as a supplement to morphologic imaging for the evaluation of tumor response to anticancer therapy in patients with soft-tissue sarcomas. As cellular changes are expected to precede morphologic changes in treated tumors, DWI performed at an early stage of fractionated therapy may provide unique prognostic information of its effectiveness. PMID- 18425833 TI - MR-guided core biopsy with MR fluoroscopy using a short, wide-bore 1.5-Tesla scanner: feasibility and initial results. AB - PURPOSE: To evaluate MR fluoroscopy in a short, wide-bore 1.5T MRI suitable for near real-time biopsy guidance. MATERIALS AND METHODS: A total of eight consecutive patients underwent MR-guided core biopsy in a 1.5T system with a 70 cm bore diameter. A total of five biopsies were performed in focal liver lesions, three biopsies in soft-tissue tumors. Before biopsy, three different fast MR sequences were compared for image quality (anatomical visibility, lesion visibility, and artifacts), and signal-to-noise ratio (SNR) and contrast-to-noise ratio (CNR) were calculated. In all cases, an MR-compatible guidance needle was positioned under MR fluoroscopy using the most suitable sequence. RESULTS: In each patient the guidance needle could be placed accurately under MR fluoroscopy without having to remove the patient from the isocenter of the magnet during needle movement. All biopsies were technically successful and appropriate specimens could be obtained. In prebiopsy imaging, a T2-weighted single shot turbo spinecho sequence (half-Fourier acquisition single-shot turbo spin-echo [HASTE]) achieved the best rating for lesion visibility and superior SNR and CNR values. CONCLUSION: Findings of this study demonstrate that MR fluoroscopy for biopsy guidance in a short, wide-bore 1.5T scanner is feasible. This scanner combines the patient access advantages of an open-bore system with the superior image quality and speed of a high-field scanner. In our series, the HASTE sequence was best suited for MR-guidance of biopsies. PMID- 18425834 TI - Automatic repositioning of MRSI voxels in longitudinal studies: impact on reproducibility of metabolite concentration measurements. AB - PURPOSE: To study an automatic repositioning method to reduce variability in longitudinal MRSI exams based on a priori image registration. Longitudinal proton MR spectroscopic imaging ((1)H MRSI) exams to study the effects of disease or treatment are becoming increasingly common. However, one source of variability in such exams arises from imperfect relocalization of the MRSI grid in the follow-up exams. MATERIALS AND METHODS: Six healthy subjects were each scanned three times during the course of 1 day. In each follow-up exam a manually placed MRSI grid was acquired in addition to the automatically repositioned MRSI grid. Then coefficients of variance between baseline and follow-up scans were calculated for N-acetylaspartate, creatine, and choline. In addition, the overall MRSI grid overlap and individual voxel overlaps were also calculated for both the visually and automatically repositioned voxels. RESULTS: Streamlined workflow, reduced variability of metabolite concentration measurements, and increased voxel overlaps are noted when this automatic repositioning procedure is compared to the visual MRSI grid repositioning approach. CONCLUSION: Our results suggest that this approach is able to improve reproducibility in longitudinal MRS exams. PMID- 18425835 TI - Delayed enhancement in hypertrophic cardiomyopathy: comparison with myocardial tagging MRI. AB - PURPOSE: To evaluate the relationship between delayed enhancement (DE) and regional left ventricular function in hypertrophic cardiomyopathy (HCM) using gadolinium enhancement MRI and myocardial tagging MRI. MATERIALS AND METHODS: Cine imaging, delayed enhancement imaging, and tagging MRI were performed in 25 patients with HCM. The location, pattern, and extent of DE were evaluated. Circumferential shortening (Ecc) was obtained by analyzing MR tagging images with HARP software. RESULTS: DE occurred in 21 (84%) patients with a high frequency of localization in the septum and the right ventricular attachment sites. Circumferential shortening was significantly decreased in the enhanced segments compared with nonenhanced segments (P < 0.0001). The myocardial wall was thicker in the enhanced segments than in the nonenhanced segments (P < 0.0001). However, circumferential shortening was significantly decreased in the enhanced segments of the same thickness (P < 0.0001). Circumferential shortening was more substantially impaired in the segments with focal nodular enhancement than those in the segments with ill-defined patchy enhancement (P = 0.0002). CONCLUSION: In HCM, DE is commonly found and circumferential shortening is significantly impaired in the regions with DE, regardless of the degree of myocardial hypertrophy. Focal nodular enhancement is particularly related with regional dysfunction in patients with HCM. PMID- 18425836 TI - Magnetization transfer based contrast for imaging denatured collagen. AB - PURPOSE: To study of the sensitivity of various NMR and MRI methods and parameters to the degree of thermal denaturation of collagen. MATERIALS AND METHODS: Collagen type I powder was washed with methanol:chloroform to remove traces of lipids and then suspended in saline. Denaturation was carried out by heating the suspension for 5-120 minutes at a temperature range of 50-100 degrees C. The NMR methods tested were two T(2) filter methods: Goldman-Shen (GS) and Edzes-Samulski (ES); magnetization transfer contrast (MTC); double quantum filtering (DQF) and high resolution spectroscopy. MRI contrasts based on these methods were compared. RESULTS: The following parameters were found to be sensitive to denaturation of collagen: 1) the amount of spins that experience high dipolar interactions as assessed by DQF; 2) MTR; 3) k(w)T(1w) (where k(w) is the magnetization transfer rate from water to collagen, and T(1w) is the water protons longitudinal relaxation time); and 4) aliphatic residues content. The contrast between native and denatured collagen was improved by all the tested methods, with ES and DQF producing the highest contrast. CONCLUSION: Methods depending on T(2) filtering and DQF were found to be sensitive to the degree of thermal denaturation of collagen and improve the contrast between native and denatured collagen. PMID- 18425837 TI - Contribution of cardiac-induced brain pulsation to the noise of the diffusion tensor in Turboprop diffusion tensor imaging (DTI). AB - PURPOSE: To assess the effects of cardiac-induced brain pulsation on the noise of the diffusion tensor in Turboprop (a form of periodically rotated overlapping parallel lines with enhanced reconstruction [PROPELLER] imaging) diffusion tensor imaging (DTI). MATERIALS AND METHODS: A total of six healthy human subjects were imaged with cardiac-gated as well as nongated Turboprop DTI. Gated and nongated Turboprop DTI datasets were also simulated using actual data acquired exclusively during the diastolic or systolic period of the cardiac cycle. The total variance of the diffusion tensor (TVDT) was measured and compared between acquisitions. RESULTS: The TVDT near the ventricles was significantly reduced in cardiac-gated compared to nongated Turboprop DTI acquisitions. Furthermore, the effects of brain pulsation were reduced, but not eliminated, when increasing the amount of data collected. Finally, data corrupted by cardiac-induced pulsation were not consistently detected by the step of the conventional Turboprop reconstruction algorithm that evaluates the quality of data in different blades. Thus, the inherent quality weighting of the conventional Turboprop reconstruction algorithm was unable to compensate for the increased noise in the diffusion tensor due to brain pulsation. CONCLUSION: Cardiac-induced brain pulsation increases the TVDT in Turboprop DTI. Use of cardiac gating to limit data acquisition to the diastolic period of the cardiac cycle reduces the TVDT at the expense of imaging time. PMID- 18425838 TI - A tracking-based diffusion tensor imaging segmentation method for the detection of diffusion-related changes of the cervical spinal cord with aging. AB - PURPOSE: To compare region of interest (ROI)-based and diffusion tensor tractography (DTT)-based methods for evaluating diffusion properties of the spinal cord as a function of age. MATERIALS AND METHODS: Commonly, an ROI segmentation is used to delineate the spinal cord. In this work, new segmentation methods are developed based on DTT. In a first, DTT-based, segmentation approach, the diffusion properties are calculated on the tracts. In a second method, the diffusion properties are analyzed in the spinal cord voxels that contain a certain number of tracts. We studied the changes in diffusion properties of the human spinal cord in subjects of different ages. Diffusion tensor imaging (DTI) measurements of the cervical spinal cord were acquired on 42 healthy volunteers (age range = 19-87 years). The fractional anisotropy (FA), the mean diffusivity (MD), and eigenvalues (lambda(1), lambda(2), and lambda(3)) were compared for the ROI- and DTT-based segmentation methods. RESULTS: Our automatic techniques are shown to be highly reproducible and sensitive for detecting DTI changes. FA decreased (r = -0.38; P < 0.05), whereas MD and eigenvalues increased (r = +/- 0.45; P < 0.05) with age. These trends were not statistically significant for the ROI-based segmentation (P > 0.05). CONCLUSION: DTT is a robust and reproducible technique to segment the voxels of interest in the spinal cord. PMID- 18425839 TI - Comparison of quantitative coronary angiography and first-pass perfusion magnetic resonance imaging for the detection of an impaired coronary perfusion in nonsevere coronary stenosis. AB - PURPOSE: To compare quantitative coronary angiography (QCA) and first-pass perfusion magnetic resonance imaging (FPP-MRI) in symptomatic patients with nonsevere coronary stenosis to detect a reduced coronary flow velocity reserve (CFVR). MATERIALS AND METHODS: In 35 patients, FPP-MRI and CFVR measurements were performed in 40 coronary arteries with a diameter stenosis (DS) <70% by QCA. From FPP-MRI a myocardial perfusion reserve index (MPRI) was calculated. CFVR was calculated as the ratio of the average peak flow velocity during infusion of adenosine and at rest and was considered reduced if <2. Diagnostic performance of MPRI and DS to detect a reduced CFVR was compared by receiver operating characteristic (ROC) curve analysis. RESULTS: CFVR was reduced in 16 coronary arteries (40%). Mean DS did not differ in coronary arteries with a reduced CFVR (41.0% +/- 13.3) and a normal CFVR (36.5% +/- 12.3; P = 0.281). Mean MPRI was lower in coronary arteries with a reduced CFVR (1.12 +/- 0.12) compared to a normal CFVR (1.33 +/- 0.2; P < 0.001). Sensitivity, specificity, and area under the ROC curve (AUC) were higher for MPRI (81%, 79%, 0.84) than for DS (56%, 58%, 0.60). CONCLUSION: FPP-MRI detects impaired CFVR in symptomatic patients with nonsevere coronary stenosis more accurately than QCA and can identify patients with symptomatic ischemia. PMID- 18425840 TI - Assessment of regional myocardial oxygenation changes in the presence of coronary artery stenosis with balanced SSFP imaging at 3.0 T: theory and experimental evaluation in canines. AB - PURPOSE: To examine the dependence of steady-state free-precession (SSFP) -based myocardial blood-oxygen-level-dependent (BOLD) contrast on field strength using theoretical and experimental models. MATERIALS AND METHODS: Numerical simulations using a two-pool exchange model and a surgically prepared dog model were used to assess the SSFP-based myocardial BOLD signal changes at 1.5T and 3.0T. Experimental studies were performed in eight canines with pharmacological vasodilation under various levels of left circumflex coronary artery stenosis. Experimentally obtained BOLD signal changes were correlated against microsphere based true flow changes. RESULTS: Theoretical results showed that, at 3.0T, relative to 1.5T, a threefold increase in oxygen sensitivity can be expected. Experimental studies in canines showed near similar results-a 2.5 +/- 0.2-fold increase in BOLD sensitivity at 3.0T relative to 1.5T (P < 0.05). Based on the scatter gram of BOLD data and microsphere data, it was found that the minimum regional flow difference that can be detected with SSFP-based myocardial BOLD imaging at 1.5T and 3.0T were 2.9 and 1.6, respectively (P < 0.05). CONCLUSION: This study demonstrated that SSFP-based myocardial BOLD sensitivity is substantially greater at 3.0T compared with 1.5T. The findings here suggest that SSFP-based myocardial BOLD imaging at 3.0T may have the necessary sensitivity to detect the clinically required minimum flow difference of 2.0. PMID- 18425842 TI - Assessment of muscle oxygenation with balanced SSFP: a quantitative signal analysis. AB - PURPOSE: To investigate the feasibility of balanced steady-state free precession (b-SSFP) for blood oxygenation level-dependent (BOLD) MRI during a short-term ischemia/reactive hyperemia (RH) experiment on human calf muscles. MATERIALS AND METHODS: To investigate contributions to the b-SSFP signal during an RH experiment, the relaxation times T(1), T(2), and T(2) (*) were quantified in an interleaved fashion. Data from soleus, gastrocnemius, and tibialis muscle groups of five healthy subjects were evaluated. RESULTS: During ischemia a decreased b SSFP signal amplitude as well as a decrease in T(2), T(2) (*), and the initial intensity I(0) was observed. RH provoked an overshoot of T(2), T(2) (*), and the b-SSFP signal. No paradigm-related changes in T(1) were observed. Comparing the evolution of transverse relaxation times, initial intensity, and b-SSFP signal amplitude, we concluded that the measured b-SSFP signal in muscle tissue is not only determined by T(2) variations but also significantly influenced by I(0) changes. These I(0) changes are attributed to spin density variations since inflow effects were suppressed by saturation bands. CONCLUSION: b-SSFP signal changes during a RH paradigm cannot unambiguously be assigned to oxygenation changes. Therefore, care has to be taken with their interpretation. PMID- 18425841 TI - Detection of choline signal in human breast lesions with chemical-shift imaging. AB - PURPOSE: To investigate the application of MR spectroscopy using chemical-shift imaging (CSI) for characterizing human breast lesions at 1.5T, and to evaluate the diagnostic performance using ROC (receiver operating characteristics) analysis. MATERIALS AND METHODS: Thirty-six patients (35-73 years old, mean 52), with 27 malignant and 9 benign lesions, underwent anatomical imaging, dynamic contrast-enhanced MR imaging, and CSI. The ROC analysis was performed and the cutoff point yielding the highest accuracy was found to be a choline (Cho) signal to-noise ratio (SNR) >3.2. RESULTS: The mean Cho SNR was 2.8 +/- 0.8 (range, 1.8 4.3) for the benign group and 5.9 +/- 3.4 (2.1-17.5) for the malignant group (P = 0.01). Based on the criterion of Cho SNR >3.2 as malignant, CSI correctly diagnosed 22 of 27 malignant lesions and 7 of 9 benign lesions, resulting in a sensitivity of 81%, specificity of 78%, and overall accuracy of 81%. If the criterion was set higher at Cho SNR >4.0 the specificity improved to 89% but sensitivity was lowered to 67%. CONCLUSION: The ROC analysis presented in this work could be used to set an objective diagnostic criterion depending on preferred emphasis on sensitivity or specificity. PMID- 18425843 TI - Gadolinium-based contrast agents and their potential role in the pathogenesis of nephrogenic systemic fibrosis: the role of excess ligand. AB - PURPOSE: To investigate the role of excess ligand present in gadolinium (Gd) based contrast agents in the development of nephrogenic systemic fibrosis (NSF). Using a dosing regimen to simulate the exposure seen in patients with severe renal impairment, we investigated the effect of excess ligand on Gd-deposition and the depletion of endogenous ions. MATERIALS AND METHODS: Gadodiamide and gadoversetamide were formulated with 0%, 5%, and 10% excess ligand. Forty-two, healthy, male Hannover Wistar rats received daily intravenous injections of each formulation over a period of 20 days. At the end of the study, histopathological analysis of the skin was performed and the concentrations of Gd, Zn, and Cu were measured in several tissues. The levels of Zn in the urine were also measured. RESULTS: The most severe skin lesions were observed after injection of formulations containing 0% free ligand and in those animals with the highest Gd concentrations in the skin. There were no significant reductions in the levels of Zn or Cu observed in the skin; however, the levels of Zn in the urine were elevated following administration of formulations with the highest amount of excess ligand. CONCLUSION: Our findings suggest that there is an inverse correlation between the amount of excess ligand present in Gd-containing contrast agents and the amount of Gd in the tissue, and further underline the importance of the inherent stability of these agents in the development of NSF. PMID- 18425844 TI - Arterial spin labeling of cerebral perfusion territories using a separate labeling coil. AB - PURPOSE: To obtain cerebral perfusion territories of the left, the right, and the posterior circulation in humans with high signal-to-noise ratio (SNR) and robust delineation. MATERIALS AND METHODS: Continuous arterial spin labeling (CASL) was implemented using a dedicated radio frequency (RF) coil, positioned over the neck, to label the major cerebral feeding arteries in humans. Selective labeling was achieved by flow-driven adiabatic fast passage and by tilting the longitudinal labeling gradient about the Y-axis by theta = +/- 60 degrees . RESULTS: Mean cerebral blood flow (CBF) values in gray matter (GM) and white matter (WM) were 74 +/- 13 mL . 100 g(-1) . minute(-1) and 14 +/- 13 mL . 100 g( 1) . minute(-1), respectively (N = 14). There were no signal differences between left and right hemispheres when theta = 0 degrees (P > 0.19), indicating efficient labeling of both hemispheres. When theta = +60 degrees , the signal in GM on the left hemisphere, 0.07 +/- 0.06%, was 92% lower than on the right hemisphere, 0.85 +/- 0.30% (P < 1 x 10(-9)), while for theta = -60 degrees , the signal in the right hemisphere, 0.16 +/- 0.13%, was 82% lower than on the contralateral side, 0.89 +/- 0.22% (P < 1 x 10(-10)). Similar attenuations were obtained in WM. CONCLUSION: Clear delineation of the left and right cerebral perfusion territories was obtained, allowing discrimination of the anterior and posterior circulation in each hemisphere. PMID- 18425845 TI - Effect of intramyocardial bone marrow cell injection on diastolic function in patients with chronic myocardial ischemia. AB - PURPOSE: To evaluate the effect of intramyocardial bone marrow cell injection on diastolic function in patients with chronic myocardial ischemia. MATERIALS AND METHODS: In 24 patients (19 male; 65 +/- 9 years) with refractory angina (Canadian Cardiovascular Society [CCS] class III-IV) 84.6 +/- 28.7 x 10(6) bone marrow-derived mononuclear cells were injected intramyocardially (using the NOGA system) in regions with ischemia on Tc-99m tetrofosmin single photon emission computed tomography (SPECT). Diastolic function was evaluated at baseline and at three months using magnetic resonance imaging (MRI) and tissue Doppler imaging (TDI). RESULTS: MRI revealed an increased early (E) peak filling rate (374 +/- 121 mL/second vs. 412 +/- 102 mL/second; P = 0.04), whereas the atrial (A) peak filling rate remained unchanged (340 +/- 81 mL/second vs. 334 +/- 93 mL/second; P = not significant [NS]). The E/A peak flow ratio increased from 1.09 +/- 0.33 to 1.23 +/- 0.47 at three months (P = 0.02). TDI demonstrated a significant improvement in early diastolic velocity (E') from 4.4 +/- 1.7 cm/second to 4.8 +/ 1.6 cm/second at three months (P = 0.03), whereas the late diastolic velocity (A') remained unchanged (6.0 +/- 1.6 cm/second vs. 6.0 +/- 1.7 cm/second; P = NS). Consequently, the E'/A' ratio increased from 0.74 +/- 0.19 to 0.84 +/- 0.28 at three months (P = 0.02). CONCLUSION: Intramyocardial bone marrow cell injection in patients with chronic myocardial ischemia improved MRI and TDI derived parameters of diastolic function. PMID- 18425846 TI - Voxel-based diffusion tensor magnetic resonance imaging evaluation of low-grade hepatic encephalopathy. AB - PURPOSE: To quantify the changes in brain water diffusivity in hepatic encephalopathy (HE) associated with cirrhosis using diffusion tensor imaging (DTI) and to correlate with neuropsychological (NP) scores. MATERIALS AND METHODS: DTI was performed in 14 patients with low-grade HE and age/gender comparable 16 healthy controls. Whole brain mean diffusivity (MD) and fractional anisotropy (FA) maps were calculated, normalized to common space, smoothed, and compared voxel-by-voxel between groups using analysis of covariance with age included as a covariate. The average MD and FA values were also calculated from individual subjects for selected brain regions and correlated with the neuropsychological scores. RESULTS: Patients with HE showed increased MD in the cortical gray and white matter and the internal capsule. Less extensive brain regions with decreased FA were observed in the bilateral frontal and occipital white matter. MD values from the corpus callosum correlated inversely with several NP scores among HE patients and controls. Positive correlations were observed with FA values and cognitive scores. CONCLUSION: Voxel-based DTI analysis showed widespread brain regions with increased MD values, indicating enhanced water content and decreased FA in cirrhotic patients with HE. The MD and FA values from selected regions correlated with the NP scores. PMID- 18425847 TI - The novel N-substituted benztropine analog GA2-50 possesses pharmacokinetic and pharmacodynamic profiles favorable for a candidate substitute medication for cocaine abuse. AB - GA2-50 is a novel N-substituted benztropine analog with improved potency and selectivity for the dopamine transporter. The pharmacokinetic and pharmacodynamic properties of GA2-50 were characterized as a part of its preclinical evaluation as a substitute medication for cocaine abuse. In vitro transport and metabolism studies as well as pharmacokinetic studies in rats were conducted. Effect of GA2 50 on the extracelluar nucleus accumbens (NAc) dopamine levels and on cocaine's induced dopamine elevation was evaluated using intracerebral microdialysis. GA2 50 showed high transcellular permeability despite being a P-glycoprotein substrate. GA2-50 was a substrate of human CYP2D6, CYP2C19, CYP2E1, rat CYP2C11, CYP2D1, CYP3A1, and CYP1A2; with low intrinsic clearance values. In vivo, GA2-50 showed high brain uptake (R(i) approximately 10), large volume of distribution (V(ss) = 37 L/kg), and long elimination half-life (t((1/2)) = 19 h). GA2-50 resulted in 1.6- and 2.7-fold dopamine elevation at the 5 and 10 mg/kg i.v. doses. Dopamine elevation induced by GA2-50 was significantly reduced, slower and longer lasting than previously observed for cocaine. GA2-50 had no significant effect on cocaine's induced dopamine elevation upon simultaneous administration. Results from the present study indicate that GA2-50 possesses several attributes sought after for a substitute medication for cocaine abuse. PMID- 18425848 TI - Expression and function of the LIM homeobox containing genes Lhx3 and Lhx4 in the mouse placenta. AB - The LIM homeobox containing genes of the LIM-3 group, Lhx3 and Lhx4, are critical for normal development. Both genes are involved in the formation of the pituitary and the motoneuron system and loss of either gene causes perinatal lethality. Previous studies had shown that Lhx3 is overexpressed in hyperplastic placentas of mouse interspecies hybrids. To determine the role of LHX3 in the mouse placenta, we performed expression and function analyses. Our results show that Lhx3 exhibits specific spatial and temporal expression in the mouse placenta. However, deletion of Lhx3 does not produce a placental phenotype. To test whether this is due to functional substitution by Lhx4, we performed a phenotype analysis of Lhx3-/-; Lhx4-/- double-mutant placentas. A subset of Lhx3-/-; Lhx4-/- placentas exhibited abnormal structure of the labyrinth. However, absence of both LIM-3 genes did not interfere with placental transport nor consistently with expression of target genes such as Gnrhr. Thus, LHX3 and LHX4 appear to be dispensable for placental development and function. PMID- 18425849 TI - Chromatin remodeling during mouse and human embryonic stem cell differentiation. AB - Embryonic stem cell (ESC) differentiation is an excellent model to study chromatin changes at developmentally regulated loci. Differentiating mouse and human ESCs increase genome-wide acetylation (euchromatic) and tri-methylation (heterochromatic) of lysine 9 on histone H3. The Oct4 locus is euchromatic when expressed in undifferentiated ESCs and heterochromatic after differentiation. Brachyury T, a mesoderm-specific transcription factor, is not yet expressed in undifferentiated cells, where its locus has "bivalent" tri-methyl lysine 4 and lysine 27 modifications. During directed differentiation to pre-cardiac mesoderm, the activated brachyury locus has high levels of tri-methyl lysine 4 (euchromatin), switching to heterochromatin after gene silencing. Thus, ESC differentiation is accompanied by genome-wide commitment to euchromatin or heterochromatin. Undifferentiated hESCs bivalently modify the brachyury locus, activate it to euchromatin during mesoderm induction, and subsequently repress it to heterochromatin, demonstrating, to our knowledge, the first analysis of chromatin dynamics at a locus essential for mesoderm and endoderm differentiation. PMID- 18425850 TI - CMF1-Rb interaction promotes myogenesis in avian skeletal myoblasts. AB - CMF1 protein is expressed in developing striated muscle before the expression of contractile proteins, and depletion of CMF1 in myoblasts results in inability to express muscle-specific proteins. Previous studies of CMF1 identify a functional Rb-binding domain, which is conserved in the murine and human homologues. Here, we show that CMF1 binds Rb family members, while a CMF1 protein with deletion of the Rb-binding domain (Rb-del CMF1) does not. Myogenic cell lines over-expressing Rb-del CMF1 proliferate normally, but exhibit markedly impaired differentiation, including dramatically reduced contractile proteins gene expression and failure to fuse into myotubes. Furthermore, by quantitative real-time polymerase chain reaction, MyoD and Myf5 mRNA levels are comparable to wild-type, while myogenin and contractile protein mRNA levels are significantly attenuated. These data demonstrate that CMF1 regulates myocyte differentiation by interaction with Rb family members to induce expression of myogenic regulatory factors. PMID- 18425851 TI - GSK-3beta and alpha-catenin binding regions of beta-catenin exert opposing effects on the terminal ventral optic axonal projection. AB - We overexpressed two deletion mutants of the N-terminal domain of beta-catenin in ventral optic axons in living Xenopus tadpoles. One deletion mutant contained both the alpha-catenin and the GSK-3beta binding sites of the N-terminal domain of beta-catenin (NTERM), and the second deletion mutant contained only the GSK 3beta binding site (beta-cat107). Expression of NTERM in ventral optic axons dispersed and induced anterior and lateral shifts in their targeting locations in the dorsal tectum. In contrast, beta-cat107 compressed and shifted the synaptic targeting locations of ventral optic axons medially and posteriorly. In addition, NTERM-expressing ventral optic axons formed arbors that were wider than controls whereas beta-cat107 axonal arbors were narrower compared with controls. These data suggest that the interactions of beta-catenin with alpha-catenin and GSK 3beta exert opposing effects on the terminal projections of ventral optic axons. PMID- 18425853 TI - Novel murine clonal cell lines either express slow or mixed (fast and slow) muscle markers following differentiation in vitro. AB - We have investigated whether the phenotype of myogenic clones derived from satellite cells of different muscles from the transgenic immortomouse depended on muscle type origin. Clones derived from neonatal, or 6- to 12-week-old fast and slow muscles, were analyzed for myosin and enolase isoforms as phenotypic markers. All clones derived from slow-oxidative muscles differentiated into myotubes with a preferentially slow contractile phenotype, whereas some clones derived from rapid-glycolytic or neonatal muscles expressed both fast and slow myosin isoforms. Thus, muscle origin appears to bias myosin isoform expression in myotubes. The neonatal clone (WTt) was cultivated in various medium and substrate conditions, allowing us to determine optimized conditions for their differentiation. Matrigel allowed expressions of adult myosin isoforms, and an isozymic switch from embryonic alpha- toward muscle-specific beta-enolase, never previously observed in vitro. These cells will be a useful model for in vitro studies of muscle fiber maturation and plasticity. PMID- 18425852 TI - Spatiotemporal protein distribution of TGF-betas, their receptors, and extracellular matrix molecules during embryonic tendon development. AB - Tendon is one of the least understood tissues of the musculoskeletal system in terms of development and morphogenesis. Collagen fibrillogenesis has been the most studied aspect of tendon development, focusing largely on the role of matrix molecules such as collagen type III and decorin. While involvement of matrix molecules in collagen fibrillogenesis during chick tendon development is well understood, the role of growth factors has yet to be elucidated. This work examines the expression patterns of transforming growth factor (TGF) -beta1, beta2, and -beta3, and their receptors with respect to expression patterns of collagen type III, decorin, and fibronectin. We focus on the intermediate stages of tendon development in the chick embryo, a period during which the tendon micro and macro-architecture are being established. Our findings demonstrate for the first time that TGF-beta1, -beta2, and -beta3 have distinct spatiotemporal developmental protein localization patterns in the developing tendon and strongly suggest that these isoforms have independent roles in tendon development. PMID- 18425854 TI - Tyl1a, a TDP-6-deoxy-D-xylo-4-hexulose 3,4-isomerase from Streptomyces fradiae: structure prediction, mutagenesis and solvent isotope incorporation experiments to investigate reaction mechanism. AB - Understanding the structure and mechanism of sugar nucleotide processing enzymes is invaluable in the generation of designer enzymes for biotransformation, for instance, in connection with engineering antibiotic glycosylation. In this study, homology modelling and mechanistic comparison to the structurally related RmlC epimerase family has been used to identify and assign functions to active-site residues in the Tyl1a-catalysed keto-sugar nucleotide isomerisation process. Tyl1a His63 is implicated as the base that initiates the isomerisation process by substrate C-3 deprotonation, with Arg109 stabilising the resulting enolate. Subsequent O-3 deprotonation (potentially by His65) and C-4 protonation (potentially by Tyr49) complete the isomerisation process. PMID- 18425855 TI - How is oxygen incorporated into oxides? A comprehensive kinetic study of a simple solid-state reaction with SrTiO3 as a model material. AB - The kinetics of stoichiometry change of an oxide--a prototype of a simple solid state reaction and a process of substantial technological relevance--is studied and analyzed in great detail. Oxygen incorporation into strontium titanate was chosen as a model process. The complete reaction can be phenomenologically and mechanistically understood beginning with the surface reaction and ending with the transport in the perovskite. Key elements are a detailed knowledge of the defect chemistry of the perovskite as well as the application of a variety of experimental and theoretical tools, many of them evolving from this study. The importance of the reaction and transport steps for (electro)chemical applications is emphasized. PMID- 18425856 TI - Exploring crystal morphology of nanoporous hosts from time-dependent guest profiles. PMID- 18425857 TI - Quantitative evaluation of anion-pi interactions in solution. PMID- 18425858 TI - Cinchona alkaloid catalyzed enantioselective fluorination of allyl silanes, silyl enol ethers, and oxindoles. PMID- 18425859 TI - Interventions for enhancing medication adherence. AB - BACKGROUND: People who are prescribed self-administered medications typically take less than half the prescribed doses. Efforts to assist patients with adherence to medications might improve the benefits of prescribed medications, but also might increase their adverse effects. OBJECTIVES: To update a review summarizing the results of randomized controlled trials (RCTs) of interventions to help patients follow prescriptions for medications for medical problems, including mental disorders but not addictions. SEARCH STRATEGY: We updated searches of The Cochrane Library, MEDLINE, CINAHL, EMBASE, International Pharmaceutical Abstracts (IPA), PsycINFO (all via OVID) and Sociological Abstracts (via CSA) in January 2007 with no language restriction. We also reviewed bibliographies in articles on patient adherence and articles in our personal collections, and contacted authors of relevant original and review articles. SELECTION CRITERIA: Articles were selected if they reported an unconfounded RCT of an intervention to improve adherence with prescribed medications, measuring both medication adherence and treatment outcome, with at least 80% follow-up of each group studied and, for long-term treatments, at least six months follow-up for studies with positive initial findings. DATA COLLECTION AND ANALYSIS: Study design features, interventions and controls, and results were extracted by one review author and confirmed by at least one other review author. We extracted adherence rates and their measures of variance for all methods of measuring adherence in each study, and all outcome rates and their measures of variance for each study group, as well as levels of statistical significance for differences between study groups, consulting authors and verifying or correcting analyses as needed. The studies differed widely according to medical condition, patient population, intervention, measures of adherence, and clinical outcomes. Therefore, we did not feel that quantitative analysis was scientifically justified; rather, we conducted a qualitative analysis. MAIN RESULTS: For short term treatments, four of ten interventions reported in nine RCTs showed an effect on both adherence and at least one clinical outcome, while one intervention reported in one RCT significantly improved patient adherence, but did not enhance the clinical outcome. For long-term treatments, 36 of 81 interventions reported in 69 RCTs were associated with improvements in adherence, but only 25 interventions led to improvement in at least one treatment outcome. Almost all of the interventions that were effective for long-term care were complex, including combinations of more convenient care, information, reminders, self-monitoring, reinforcement, counseling, family therapy, psychological therapy, crisis intervention, manual telephone follow-up, and supportive care. Even the most effective interventions did not lead to large improvements in adherence and treatment outcomes. AUTHORS' CONCLUSIONS: For short-term treatments several quite simple interventions increased adherence and improved patient outcomes, but the effects were inconsistent from study to study with less than half of studies showing benefits. Current methods of improving adherence for chronic health problems are mostly complex and not very effective, so that the full benefits of treatment cannot be realized. High priority should be given to fundamental and applied research concerning innovations to assist patients to follow medication prescriptions for long-term medical disorders. PMID- 18425860 TI - Physician advice for smoking cessation. AB - BACKGROUND: Healthcare professionals frequently advise patients to improve their health by stopping smoking. Such advice may be brief, or part of more intensive interventions. OBJECTIVES: The aims of this review were to assess the effectiveness of advice from physicians in promoting smoking cessation; to compare minimal interventions by physicians with more intensive interventions; to assess the effectiveness of various aids to advice in promoting smoking cessation, and to determine the effect of anti-smoking advice on disease-specific and all-cause mortality. SEARCH STRATEGY: We searched the Cochrane Tobacco Addiction Group trials register. Date of the most recent search: September 2007. SELECTION CRITERIA: Randomized trials of smoking cessation advice from a medical practitioner in which abstinence was assessed at least six months after advice was first provided. DATA COLLECTION AND ANALYSIS: We extracted data in duplicate on the setting in which advice was given, type of advice given (minimal or intensive), and whether aids to advice were used, the outcome measures, method of randomization and completeness of follow up. The main outcome measure was abstinence from smoking after at least six months follow up. We also considered the effect of advice on mortality where long-term follow-up data were available. We used the most rigorous definition of abstinence in each trial, and biochemically validated rates where available. Subjects lost to follow up were counted as smokers. Effects were expressed as relative risks. Where possible, meta-analysis was performed using a Mantel-Haenszel fixed effect model. MAIN RESULTS: We identified 41 trials, conducted between 1972 and 2007, including over 31,000 smokers. In some trials, subjects were at risk of specified diseases (chest disease, diabetes, ischaemic heart disease), but most were from unselected populations. The most common setting for delivery of advice was primary care. Other settings included hospital wards and outpatient clinics, and industrial clinics. Pooled data from 17 trials of brief advice versus no advice (or usual care) detected a significant increase in the rate of quitting (relative risk (RR) 1.66, 95% confidence interval (CI) 1.42 to 1.94). Amongst 11 trials where the intervention was judged to be more intensive the estimated effect was higher (RR 1.84, 95% CI 1.60 to 2.13) but there was no statistical difference between the intensive and minimal subgroups. Direct comparison of intensive versus minimal advice showed a small advantage of intensive advice (RR 1.37, 95% CI 1.20 to 1.56). Direct comparison also suggested a small benefit of follow-up visits. Only one study determined the effect of smoking advice on mortality. This study found no statistically significant differences in death rates at 20 years follow up. AUTHORS' CONCLUSIONS: Simple advice has a small effect on cessation rates. Assuming an unassisted quit rate of 2 to 3%, a brief advice intervention can increase quitting by a further 1 to 3%. Additional components appear to have only a small effect, though there is a small additional benefit of more intensive interventions compared to very brief interventions. PMID- 18425861 TI - Antibiotics for acute maxillary sinusitis. AB - BACKGROUND: Expert opinions vary on the appropriate role of antibiotics for sinusitis, one of the most commonly diagnosed conditions among adults in ambulatory care. OBJECTIVES: We examined whether antibiotics are effective in treating acute sinusitis, and if so, which antibiotic classes are the most effective. SEARCH STRATEGY: We searched the Cochrane Central Register of Controlled Trials (CENTRAL) (The Cochrane Library, 2007, Issue 3); MEDLINE (1950 to May 2007) and EMBASE (1974 to June 2007). SELECTION CRITERIA: Randomized controlled trials (RCTs) comparing antibiotics with placebo or antibiotics from different classes for acute maxillary sinusitis in adults. We included trials with clinically diagnosed acute sinusitis, whether or not confirmed by radiography or bacterial culture. DATA COLLECTION AND ANALYSIS: At least two review authors independently screened search results, extracted data and quality assessed trials. Risk ratios (RR) were calculated for differences in the intervention and control groups to see whether or not the treatment was a failure. In meta-analysing the placebo-controlled studies, the data across antibiotic classes were combined. Primary outcomes were the clinical failure rates at 7 to 15 days and 16 to 60 days follow up. MAIN RESULTS: Fifty-seven studies were included in the review; six placebo-controlled studies and 51 studies comparing different classes of antibiotics. Five studies involving 631 participants provided data for comparison of antibiotics to placebo, when clinical failure was defined as a lack of cure or improvement at 7 to 15 days follow up. These studies found a slight statistical difference in favor of antibiotics, compared to placebo, with a pooled RR of 0.66 (95% confidence interval (CI) 0.44 to 0.98). However, the clinical significance of the result is equivocal, also considering that cure or improvement rate was high in both the placebo group (80%) and the antibiotic group (90%). Based on six studies, when clinical failure was defined as a lack of total cure, there was significant difference in favor of antibiotics compared to placebo with a pooled RR of 0.74 (95% CI 0.65 to 0.84) at 7 to 15 days follow up. None of the antibiotic preparations was superior to each other. AUTHORS' CONCLUSIONS: Antibiotics have a small treatment effect in patients with uncomplicated acute sinusitis in a primary care setting with symptoms for more than seven days. However, 80% of participants treated without antibiotics improve within two weeks. Clinicians need to weigh the small benefits of antibiotic treatment against the potential for adverse effects at both the individual and general population level. PMID- 18425862 TI - Cervical insemination versus intra-uterine insemination of donor sperm for subfertility. AB - BACKGROUND: Insemination with donor sperm is an option for couples for whom in vitro fertilisation (IVF) or intra-cytoplasmic sperm injection (ICSI) has been unsuccessful, couples with azoospermia and for single women or same sex couples. Insemination of sperm can be done via cervical (CI) or intra-uterine (IUI) routes. IUI has been considered potentially more effective than CI as the sperm bypasses the cervical mucus and is deposited closer to the fallopian tubes. The cost and risks of IUI may be higher because of the need for sperm preparation and the introduction of foreign material into the uterus. Donor sperm used for artificial insemination is mainly cryopreserved, due to concerns about HIV transmission. However, cycle fecundity is higher for fresh sperm. Insemination is often combined with ovulatory stimulation, with either clomiphene or gonadotrophin. There may be risks associated with these therapies, such as higher multiple pregnancy rates. OBJECTIVES: To determine whether pregnancy outcomes are improved using intra-uterine insemination in comparison to cervical insemination in women undergoing artificial insemination with donor sperm. SEARCH STRATEGY: The following databases were searched: the Cochrane Menstrual Disorders and Subfertility Group Specialised Register, CENTRAL (The Cochrane Library) , MEDLINE, EMBASE, CINAHL and the reference lists of articles retrieved. SELECTION CRITERIA: Randomised controlled trials comparing IUI with CI were included. Crossover studies were included if pre-crossover data was available. DATA COLLECTION AND ANALYSIS: Study quality assessment and data extraction were carried out independently by two review authors (DB, JM). Authors of studies that potentially met the inclusion criteria were contacted, where possible if additional information was needed. MAIN RESULTS: The search strategy found 232 articles. Fifteen studies potentially met the inclusion criteria. Four studies were included in this review. All the included studies used cryopreserved sperm in stimulated cycles. In two studies 134 women had gonadotrophin-stimulated cycles and in two studies 74 women had clomiphene-stimulated cycles. The evidence showed that IUI after 6 cycles significantly improved live birth rates (odds ratio (OR) 1.98, 95% confidence interval (CI) 1.02 to 3.86) and pregnancy rates (OR 3.37, 95% CI 1.90 to 5.96) in comparison to cervical insemination. There was no statistically significant evidence of an effect on multiple pregnancies (OR 2.19, 95% CI 0.79 to 6.07) or miscarriages (relative risk (RR) 3.92, 95% CI 0.85 to 17.96). AUTHORS' CONCLUSIONS: The findings of this systematic review support the use of IUI rather than CI in stimulated cycles using cryopreserved sperm for donor insemination. PMID- 18425863 TI - Vaccines for preventing influenza in people with asthma. AB - BACKGROUND: Influenza vaccination is recommended for asthmatic patients in many countries as observational studies have shown that influenza infection can be associated with asthma exacerbations, but influenza vaccination itself has the potential to adversely affect pulmonary function. A recent overview concluded that there was no clear benefit of influenza vaccination in patients with asthma but this conclusion was not based on a systematic search of the literature. OBJECTIVES: Whilst influenza may cause asthma exacerbations, there is controversy about the use of influenza vaccinations, since they may precipitate an asthma attack in some people. The objective of this review was to assess the efficacy of influenza vaccination in children and adults with asthma. SEARCH STRATEGY: We searched the Cochrane Airways Group trials register and checked reference lists of articles. The last search was carried out in September 2007. SELECTION CRITERIA: Randomised trials of influenza vaccination in children (over two years of age) and adults with asthma. Studies involving people with chronic obstructive pulmonary disease were excluded. DATA COLLECTION AND ANALYSIS: Inclusion criteria and assessment of trial quality were applied by two reviewers independently. Data extraction was done by two reviewers independently. Study authors were contacted for missing information. MAIN RESULTS: Nine trials were initially included. Four of these trials were of high quality. Six further articles have been included in three updates (Bueving 2003; Castro 2001; Fleming 2006; Redding 2002; Reid 1998). The included studies covered a wide diversity of people, settings and types of influenza vaccination, but data from the more recent studies that used similar vaccines have been pooled. BENEFITS: Bueving 2003 studied 696 children with asthma and did not demonstrate a significant reduction in influenza related asthma exacerbations (Risk Difference 0.01; 95% confidence interval -0.02 to 0.04). HARMS: The pooled results of two trials involving 2306 people with asthma did not demonstrate a significant increase in asthma exacerbations in the two weeks following influenza vaccination (Risk Difference 0.00; 95% confidence interval -0.02 to 0.02). AUTHORS' CONCLUSIONS: Uncertainty remains about the degree of protection vaccination affords against asthma exacerbations that are related to influenza infection. Evidence from recently published trials indicates that there is no significant increase in asthma exacerbations immediately after vaccination (at least with inactivated influenza vaccination). There is concern regarding possible increased wheezing and hospital admissions in infants given live intranasal vaccination. PMID- 18425864 TI - Life skills programmes for chronic mental illnesses. AB - BACKGROUND: Most people with schizophrenia have a cyclical pattern of illness characterised by remission and relapses. The illness can reduce the ability of self-care and functioning and can lead to the illness becoming chronic and disabling. Rehabilitation is one of the important parts of treatments. Life skills programmes, emphasising the needs associated with independent functioning, are often a part of the rehabilitation process. These programmes, therefore, have been developed to enhance independent living and the quality of life for people with schizophrenia living in the community. OBJECTIVES: To review the effectiveness of life skills programmes with standard care or other comparable therapies for people with chronic mental health problems. SEARCH STRATEGY: We searched the Cochrane Schizophrenia Group Trials Register (May 2007) which is based on regular searches of BIOSIS, CENTRAL, CINAHL, EMBASE, MEDLINE and PsycINFO. Hand searches and scrutiny of references supplemented this process. We inspected references of all identified studies for further trials. SELECTION CRITERIA: We included all relevant randomised or quasi-randomised controlled trials for life skills programmes versus other comparable therapies or standard care involving people with serious mental illnesses. DATA COLLECTION AND ANALYSIS: We extracted data independently. For dichotomous data we calculated relative risks (RR) and their 95% confidence intervals (CI) on an intention-to treat basis, based on a random effects model. We calculated numbers needed to treat/harm (NNT/NNH) where appropriate. For continuous data, we calculated weighted mean differences (WMD) again based on a random effects model. MAIN RESULTS: We included four randomised controlled trials with a total of 318 participants. These evaluated life skills programmes versus standard care, or support group. We found no significant difference in life skills performance between people given life skills training and standard care (Patterson 2003, n=32, WMD -1.10 CI -7.8 to 5.6). Life skills training did not improve or worsen study retention (n=60, 2 RCTs, RR 1.16 CI 0.4 to 3.4). We found no significant difference in PANSS positive, negative or total scores between life skills intervention and standard care. Depression scores (HAM-D) did not reveal any significant difference between groups (Patterson 2003, n=32, WMD -0.70 CI -4.1 to 2.7). We found quality of life scores to be equivocal between participants given life skills training (Patterson 2003, n=32, WMD -0.02 CI -0.1 to 0.03) and standard care. Life skills compared with support groups also did not reveal any significant differences in PANSS scores, quality of life, or social performance skills (Patterson 2006, n=158, WMD -0.90 CI -3.4 to 1.6). AUTHORS' CONCLUSIONS: Currently there is no good evidence to suggest life skills programmes are effective for people with chronic mental illnesses. More robust data are needed from studies that are adequately powered to determine whether life skills training is beneficial for people with chronic mental health problems. PMID- 18425865 TI - Barrier agents for adhesion prevention after gynaecological surgery. AB - BACKGROUND: Pelvic adhesion can form as a result of inflammation, endometriosis or surgical trauma. During pelvic surgery, strategies to reduce pelvic adhesion formation may include placing synthetic barrier agents such as oxidised regenerated cellulose, polytetrafluoroethylene or Fibrin sheets between the pelvic structures. OBJECTIVES: To assess the effect of physical barriers used during pelvic surgery in women of reproductive age on pregnancy rates, pelvic pain, or postoperative adhesion reformation. SEARCH STRATEGY: We searched the Cochrane Menstrual Disorders and Subfertility Group Trials Register (searched September 2007) which is based on regular searches of MEDLINE, EMBASE, CINAHL, PsycINFO and CENTRAL, plus handsearching of 20 relevant journals and conference proceedings, and searches of several key grey literature sources. In addition, companies were contacted for unpublished trials. SELECTION CRITERIA: Any randomised controlled trials (RCTs) comparing the use of physical barriers versus no treatment or other physical barriers in the prevention of adhesions in women undergoing gynaecological surgery. DATA COLLECTION AND ANALYSIS: Review authors assessed trial eligibility and quality. MAIN RESULTS: Sixteen RCTs were included. Five trials randomised patients while the remainder randomised pelvic organs. Laparoscopy (six trials) and laparotomy (10 trials) were the primary surgical techniques. Indications for surgery included myomectomy (five trials), ovarian surgery (five trials), pelvic adhesions (four trials), endometriosis (one trial), and mixed (one trial). Eleven trials assessed Interceed versus no treatment, two assessed Interceed versus Gore-Tex, one trial assessed Gore-Tex versus no treatment, and one trial assessed Seprafilm versus no treatment. A single trial assessed Fibrin sheet versus no treatment. No studies reported pregnancy or reduction in pain as outcomes. The use of Interceed was associated with reduced incidence of pelvic adhesion formation, both new formation and reformation following laparoscopic surgery or laparotomy. However, this result should be interpreted with caution. Gore-Tex was more effective than no barrier or Interceed in preventing adhesion formation. There was only limited evidence that Seprafilm was effective in preventing adhesion formation following myomectomy and no evidence to support Fibrin sheet. AUTHORS' CONCLUSIONS: The absorbable adhesion barrier Interceed reduces the incidence of adhesion formation following laparoscopy and laparotomy, but there are insufficient data to support its use to improve pregnancy rates. Gore-Tex may be superior to Interceed in preventing adhesion formation but its usefulness is limited by the need for suturing and later removal. There was no evidence of effectiveness of Seprafilm and Fibrin sheet in preventing adhesion formation. PMID- 18425866 TI - Corticosteroids for the prevention and treatment of post-extubation stridor in neonates, children and adults. AB - BACKGROUND: Post-extubation stridor may prolong length of stay in the intensive care unit, particularly if airway obstruction is severe and re-intubation proves necessary. Corticosteroids, however, may be associated with adverse effects ranging from hypertension to hyperglycemia, and a more systematic assessment of the efficacy of this therapy is indicated prior to widespread adoption of this practice. OBJECTIVES: To determine whether corticosteroids are effective in preventing or treating post-extubation stridor in critically ill infants, children, or adults. SEARCH STRATEGY: We searched the Cochrane Central Register of Controlled Trials, MEDLINE, EMBASE, CINAHL and reference lists of articles. The most recent searches were conducted in April 2007. SELECTION CRITERIA: Randomized controlled trial comparing administration of corticosteroids by any route with placebo in infants, children, or adults receiving mechanical ventilation via an endotracheal tube in an intensive care unit. DATA COLLECTION AND ANALYSIS: Three review authors independently assessed trial quality and extracted data. MAIN RESULTS: Ten trials involving 2230 people were included: five in adults, two in neonates, three in children. All but one examined use of steroids for the prevention of post-extubation stridor; the remaining one concerned treatment of existing post-extubation stridor in children. Patients were drawn from heterogeneous medical/surgical populations. Dexamethasone given intravenously at least once prior to extubation was the most common steroid regimen utilized (uniformly in neonates and children). In neonates the two studies found heterogeneous results, but there was an overall non significant reduction in post extubation stridor (RR 0.42; 95% CI 0.07 to 2.32). This decrease was seen only in the study on high-risk patients treated with multiple doses of steroids around the time of extubation. In children, the two studies were clinically heterogeneous. One study included children with underlying airway abnormalities and the other excluded this group. Prophylactic corticosteroids tended to reduce reintubation and significantly reduced post-extubation stridor in the study that included children with underlying airway abnormalities (N = 62) but not in the study that excluded these children (N = 153). In five adult studies (total N = 1873), there was a non significant trend for prophylactic corticosteroid administration to reduce the risk of re-intubation (RR 0.47; 95% CI 0.16 to 1.39) and post extubation stridor (RR 0.49; 95% CI 0.20 to 1.19). These reductions were largely due to two studies that utilized repeated doses of methylprednisolone 12 to 24 hours prior to extubation. Side effects were uncommon and could not be aggregated. AUTHORS' CONCLUSIONS: Using corticosteroids to prevent (or treat) stridor after extubation has not proven effective for neonates, children or adults. However, given the consistent trends towards benefit, this intervention does merit further study. PMID- 18425867 TI - WITHDRAWN: Antibiotics for persistent nasal discharge (rhinosinusitis) in children. AB - BACKGROUND: Nasal discharge (rhinosinusitis) is extremely common in children. It is the result of inflammation of the mucosa of the upper respiratory tract, and is usually due to either infection or allergy. Infections may be caused by bacteria. OBJECTIVES: To determine the effectiveness of antibiotics versus placebo or standard therapy in treating children with persistent nasal discharge (rhinosinusitis) for at least 10 days. SEARCH STRATEGY: In this updated review, we searched the Cochrane Central Register of Controlled Trials (CENTRAL) (The Cochrane Library Issue 1, 2005) which includes the ARI Group's specialised trials register; MEDLINE (1966 to April Week 3, 2005) EMBASE (1997 to December 2004), and the references of relevant articles were searched. Authors and pharmaceutical companies were contacted. SELECTION CRITERIA: All randomised controlled trials that compared antibiotics versus placebo or standard therapy. Trials which included the use of other medications were included if all participants were allowed equal access to such medications or if the additional or alternative therapies were regarded as ineffective. Trials that only combined or compared antibiotics with surgery, or sinus puncture and lavage, were not included in the review. DATA COLLECTION AND ANALYSIS: Data were extracted by a single author for the following eight outcomes: overall clinical failure (primary outcome), failure to cure, failure to improve, clinical improvement, time to resolution, complications, side-effects and bacteriologic failure. For the dichotomous outcome variables of each individual study, proportional and absolute risk reductions were calculated using a modified intention-to-treat analysis. The summary weighted risk ratio and 95% confidence interval (CI) (fixed effect model) were calculated using the inverse of the variance of each study result for weighting (Cochrane statistical package, RevMan version 4.2). MAIN RESULTS: A total of six studies involving 562 children compared antibiotics with placebo or standard therapy. All studies were randomised but most were still susceptible to bias. Five of the studies were conducted in emergency, allergy or ENT clinics. Four of the studies required children to have x-ray changes consistent with sinusitis. Only the primary outcome (overall clinical failure) was reported in all studies. Around 40% of all randomised children did not have a clinical success documented when reviewed two to six weeks after randomisation. The control event rate varied from to 22 to 71% (mean 46%). The risk ratio estimated using a fixed effects model was 0.75 (95% CI 0.61 to 0.92). There was no evidence of statistical heterogeneity. Side effects (sufficient to cease treatment) occurred in 4 of 189 control group children (four studies). More children treated with antibiotics had side effects (17 of 330), but this difference was not statistically significant (RR 1.75, 95% CI 0.63 to 4.82). AUTHORS' CONCLUSIONS: For children with persistent nasal discharge or older children with radiographically confirmed sinusitis, the available evidence suggests that antibiotics will reduce the probability of persistence in the short to medium term. The benefits appear to be modest and around eight children must be treated in order to achieve one additional cure (number needed to treat (NNT) 8, 95% CI 5 to 29). No long term benefits have been documented. These conclusions are based on a small number of small randomised controlled trials and may require revision as additional data become available. PMID- 18425868 TI - House dust mite control measures for asthma. AB - BACKGROUND: The major allergen in house dust comes from mites. Chemical, physical and combined methods of reducing mite allergen levels are intended to reduce asthma symptoms in people who are sensitive to house dust mites. OBJECTIVES: To assess the effects of reducing exposure to house dust mite antigens in the homes of people with mite-sensitive asthma. SEARCH STRATEGY: PubMed and The Cochrane Library (last searches Nov 2007), reference lists. SELECTION CRITERIA: Randomised trials of mite control measures vs placebo or no treatment in people with asthma known to be sensitive to house dust mites. DATA COLLECTION AND ANALYSIS: Two authors applied the trial inclusion criteria and evaluated the data. Trial authors were contacted to clarify information. MAIN RESULTS: Fifty-four trials (3002 patients) were included. Thirty-six trials assessed physical methods (26 mattress encasings), 10 chemical methods, and 8 a combination of chemical and physical methods. Despite the fact that many trials were of poor quality and would be expected to exaggerate the reported effect, we did not find an effect of the interventions. For the most frequently reported outcome, peak flow in the morning (1565 patients), the standardised mean difference was 0.00 (95% confidence interval (CI) -0.10 to 0.10). There were no statistically significant differences either in number of patients improved (relative risk 1.01, 95% CI 0.80 to 1.27), asthma symptom scores (standardised mean difference -0.04, 95% CI 0.15 to 0.07), or in medication usage (standardised mean difference -0.06, 95% CI -0.18 to 0.07). AUTHORS' CONCLUSIONS: Chemical and physical methods aimed at reducing exposure to house dust mite allergens cannot be recommended. It is doubtful whether further studies, similar to the ones in our review, are worthwhile. If other types of studies are considered, they should be methodologically rigorous and use other methods than those used so far, with careful monitoring of mite exposure and relevant clinical outcomes. PMID- 18425869 TI - Enteral tube feeding for cystic fibrosis. AB - BACKGROUND: Enteral tube feeding is routinely used in many cystic fibrosis centres when weight for height percentage is less than 85%, when there has been weight loss for longer than a two-month period or when there has been no weight gain for two to three months (under five years old) or for six months (over five years old). OBJECTIVES: To examine the evidence that in people with cystic fibrosis supplemental enteral tube feeding improves nutritional status, respiratory function, and quality of life without significant adverse effects. SEARCH STRATEGY: We searched the Cochrane Cystic Fibrosis and Genetic Disorders Group Trials Register which comprises references identified from comprehensive electronic database searches and handsearches of relevant journals and abstract books of conference proceedings. We also contacted the companies that market enteral feeds and reviewed their databases. Date of the most recent search of the Group's Cystic Fibrosis Trials Register: November 2007. SELECTION CRITERIA: All randomised controlled trials comparing supplemental enteral tube feeding for one month or longer with no specific intervention in people with cystic fibrosis. DATA COLLECTION AND ANALYSIS: Thirteen trials were identified by the search; however, none were eligible for inclusion in this review. MAIN RESULTS: There are no trials included in this review. AUTHORS' CONCLUSIONS: Supplemental enteral tube feeding is widely used throughout the world to improve nutritional status in people with cystic fibrosis. The methods mostly used, nasogastric or gastrostomy feeding, are invasive, expensive, and may have a negative effect on self-esteem and body image. Reported use of enteral tube feeding suggests that it results in nutritional and respiratory improvement and it is disappointing that their efficacy has not been fully assessed by randomised controlled trials. With the more frequent recommendations to use enteral tube feeding as an early rather than a late intervention, this systematic review identifies the need for a multicentre, randomised controlled trial assessing both efficacy and possible adverse effects of enteral tube feeding in cystic fibrosis. There are no trials included in the review and we have not identified any relevant trials up to November 2007. We therefore do not plan to update this review until new trials are published. PMID- 18425870 TI - Slow advancement of enteral feed volumes to prevent necrotising enterocolitis in very low birth weight infants. AB - BACKGROUND: The major modifiable risk factors for necrotising enterocolitis in very low birth weight infants relate to enteral feeding regimens. Observational studies suggest that conservative feeding regimens such as delaying the introduction of enteral feeds or slowly advancing feed volumes reduce the risk of necrotising enterocolitis OBJECTIVES: To determine the effect of slow rates of enteral feed advancement on the incidence of necrotising enterocolitis, mortality and other morbidities in very low birth weight infants. SEARCH STRATEGY: The standard search strategy of the Cochrane Neonatal Group was used. Searches were made of the Cochrane Central Register of Controlled Trials (CENTRAL, The Cochrane Library, Issue 4, 2007), MEDLINE (1966 - December 2007), EMBASE (1980 - December 2007), CINAHL (1982- December 2007), conference proceedings, and previous reviews. SELECTION CRITERIA: Randomised or quasi-randomised controlled trials that assessed the effect of slow (up to 24 ml/kg/day) versus faster rates of advancement of enteral feed volumes upon the incidence of necrotising enterocolitis in very low birth weight infants. DATA COLLECTION AND ANALYSIS: The standard methods of the Cochrane Neonatal Group were used, with separate evaluation of trial quality and data extraction by two authors. Data were synthesised using a fixed effects model and reported using typical relative risk, typical risk difference and weighted mean difference. MAIN RESULTS: Three randomised controlled trials in which a total of 396 infants participated were identified. Few participants were extremely low birth weight or growth restricted. The trials were generally of good methodological quality but caregivers and investigators were aware of the allocated interventions. Meta analyses did not detect statistically significant effects on the risk of necrotising enterocolitis [typical relative risk 0.96 (95% confidence interval 0.48 to 1.92); typical risk difference 0.00 (95% confidence interval -0.05 to 0.05)] or all cause mortality [typical relative risk 1.40 (95% confidence interval 0.71 to 2.80); typical risk difference 0.03 (95% confidence interval 0.03 to 0.10)]. Infants who had slow rates of feed volume advancement took longer to regain birth weight [reported median difference between two and five days] and to establish full enteral feeding [reported median difference between three and five days]. No statistically significant effect on the total duration of hospital stay was detected. AUTHORS' CONCLUSIONS: The currently available data do not provide evidence that slow advancement of enteral feed volumes reduces the risk of necrotising enterocolitis in very low birth weight infants. Increasing the volume of enteral feeds at slow rather than faster rates results in several days delay in regaining birth weight and establishing full enteral feeds but the long term clinical importance of these effects is unclear. Further randomised controlled trials are needed to determine how the rate of daily increment in enteral feed volumes affects important clinical outcomes in very low birth weight infants, and particularly in extremely low birth weight or growth restricted infants. PMID- 18425871 TI - Interventions for emergency contraception. AB - BACKGROUND: Emergency contraception is using a drug or copper intrauterine device (Cu-IUD) to prevent pregnancy shortly after unprotected intercourse. Several interventions are available for emergency contraception. Information on the comparative efficacy, safety and convenience of these methods is crucial for reproductive health care providers and the women they serve. OBJECTIVES: To determine which emergency contraceptive method following unprotected intercourse is the most effective, safe and convenient to prevent pregnancy. SEARCH STRATEGY: The search included the Cochrane Controlled Trials Register, Popline, MEDLINE, PubMed, Biosis/Embase, Chinese biomedical databases and UNDP/UNFPA/WHO/World Bank Special Programme on Human Reproduction (HRP) emergency contraception database (December 2006). Content experts and pharmaceutical companies were contacted. SELECTION CRITERIA: Randomised controlled trials and controlled clinical trials including women attending services for emergency contraception following a single act of unprotected intercourse were eligible. DATA COLLECTION AND ANALYSIS: Data on outcomes and trial characteristics were extracted in duplicate and independently by two reviewers. Quality assessment was also done by two reviewers independently. Meta-analysis results are expressed as relative risk (RR) using a fixed-effects model with 95% confidence interval (CI). In the presence of statistically significant heterogeneity a random-effect model was applied. MAIN RESULTS: Eighty-one trials with 45,842 women were included. Most trials were conducted in China (70/81). There were more pregnancies with levonorgestrel compared to mid-dose (25-50 mg) (15 trials, RR: 2.01; 95% CI: 1.27 to 3.17) or low-dose mifepristone (<25 mg) (9 trials, RR: 1.43; 95% CI: 1.02 to 2.01). Low dose mifepristone was less effective than mid-dose (20 trials, RR:0.67; 95% CI: 0.49 to 0.92), but this effect was no longer statistically significant when only high quality trials were considered (6 trials, RR: 0.75; 95% CI: 0.50 to 1.10). Single dose levonorgestrel (1.5 mg) administration seemed to have similar effectiveness as the standard 12 hours apart split-dose (0.75 mg twice) (2 trials, 3830 women; RR: 0.77, 95% CI: 0.45 to 1.30). Levonorgestrel was more effective than the Yuzpe regimen in preventing pregnancy (2 trials, RR: 0.51; 95% CI: 0.31 to 0.83). CDB-2914 (a second-generation progesterone receptor modulator) may be as effective as levonorgestrel (1 trial, 1549 women; RR:1.89; 95% CI: 0.75 to 4.64) but the confidence interval is wide and the result compatible with higher or lower effectiveness. Delay in the onset of subsequent menses was the main unwanted effect of mifepristone and seemed to be dose-related. AUTHORS' CONCLUSIONS: Mifepristone middle dose (25-50 mg) was superior to other hormonal regimens. Mifepristone low dose (<25 mg) could be more effective than levonorgestrel 0.75 mg (two doses) but this was not conclusive. Levonorgestrel proved more effective than the Yuzpe regimen. The copper IUD was another effective emergency contraceptive that can provide ongoing contraception. PMID- 18425872 TI - Naftidrofuryl for intermittent claudication. AB - BACKGROUND: Lifestyle changes and cardiovascular prevention measures are a primary treatment for intermittent claudication (IC). Symptomatic treatment with vasoactive agents (Anatomic Therapeutic Chemical Classification (ATC) for medicines from the World Health Organisation class CO4A) is controversial. OBJECTIVES: To evaluate evidence on the efficacy and safety of oral naftidrofuryl (ATC CO4 21) versus placebo on the pain-free walking distance (PFWD) of people with IC by using a meta-analysis based on individual patient data (IPD). SEARCH STRATEGY: The Cochrane Peripheral Vascular Diseases Group searched their Trials Register (last searched December 2007) and CENTRAL (last searched 2007, Issue 4). We searched MEDLINE, EMBASE, International Pharmaceutical Abstracts, the Science Citation Index and contacted the authors and checked the reference lists of retrieved articles. We asked the manufacturing company for IPD. SELECTION CRITERIA: We included only randomized controlled trials (RCTs) with low or moderate risk of bias for which the IPD were available. DATA COLLECTION AND ANALYSIS: We collected data from the electronic data file or from the case report form and checked the data by a statistical quality control procedure. All randomized patients were analyzed following the intention-to-treat (ITT) principle. The geometric mean of the relative improvement in PFWD was calculated for both treatment groups in all identified studies. The effect of the drug was assessed compared with placebo on final walking distance (WDf) using multilevel and random-effect models and adjusting for baseline walking distance (WD0). For the responder analysis, therapeutic success was defined as an improvement of walking distance of at least 50%. MAIN RESULTS: We included seven studies in the IPD (n = 1266 patients). One of these studies (n = 183) was only used in the sensitivity analysis so that the main analysis included 1083 patients. The ratio of the relative improvement in PFWD (naftidrofuryl compared with placebo) was 1.37 (95% confidence interval (CI) 1.32 to 1.51, P < 0.001). The absolute difference in responder rate, or proportion successfully treated, was 22.3% (95% CI 17.1% to 27.6%). The calculated number needed to treat was 4.5 (95% CI 3.6 to 5.8). AUTHORS' CONCLUSIONS: Naftidrofuryl has a statistically significant and clinically meaningful effect of improving walking distance in the six months after initiation of therapy for people with intermittent claudication. Access by researchers to data from RCTs that is suitable for IPD analysis should be possible through repositories of data from pharmacological trials. Regular formal appraisal of the balance of risk and benefit is needed for older pharmaceutical products. PMID- 18425873 TI - Emergency intubation for acutely ill and injured patients. AB - BACKGROUND: Emergency intubation has been widely advocated as a life saving procedure in severe acute illness and injury associated with real or potential compromises to the patient's airway and ventilation. However, some initial data have suggested a lack of observed benefit. OBJECTIVES: To determine in acutely ill and injured patients who have real or anticipated problems in maintaining an adequate airway whether emergency endotracheal intubation, as opposed to other airway management techniques, improves the outcome in terms of survival, degree of disability at discharge or length of stay and complications occurring in hospital. SEARCH STRATEGY: We searched the Cochrane Injuries Group Specialised Register (December 2006), Cochrane Central Register of Controlled Trials (CENTRAL) (The Cochrane Library 2006, Issue 4), MEDLINE (1950 to November 2006), EMBASE (1980 to week 50, December 2006), National Research Register (Issue 4, 2006), CINAHL (1980 to December 2006), BIDS (to December 2006) and ICNARC (to December 2006). We also examined reference lists of articles for relevant material and contacted experts in the field. Non-English language publications were searched for and examined. SELECTION CRITERIA: All randomised (RCTs) or controlled clinical trials involving the emergency use of endotracheal intubation in the injured or acutely ill patient were examined. DATA COLLECTION AND ANALYSIS: The full texts of 452 studies were reviewed independently by two authors using a standard form. Where the review authors felt a study may be relevant for inclusion in the final review or disagreed, the authors examined the study and a collective decision was made regarding its inclusion or exclusion from the review. The results were not combined in a meta-analysis due to the heterogeneity of patients, practitioners and alternatives to intubation that were used. MAIN RESULTS: We identified three eligible RCTs carried out in urban environments. Two trials involved adults with non-traumatic out-of-hospital cardiac arrest. One of these trials found a non-significant survival disadvantage in patients randomised to receive a physician-operated intubation versus a combi tube (RR 0.44, 95% CI 0.09 to 1.99). The second trial detected a non-significant survival disadvantage in patients randomised to paramedic intubation versus an oesophageal gastric airway (RR 0.86, 95% CI 0.39 to 1.90). The third included study was a trial of children requiring airway intervention in the prehospital environment. The results indicated no difference in survival (OR 0.82, 95% CI 0.61 to 1.11) or neurologic outcome (OR 0.87, 95% CI 0.62 to 1.22) between paramedic intubation versus bag-valve-mask ventilation and later hospital intubation by emergency physicians; however, only 42% of the children randomised to paramedic endotracheal intubation actually received it. AUTHORS' CONCLUSIONS: The efficacy of emergency intubation as currently practised has not been rigorously studied. The skill level of the operator may be key in determining efficacy. In non-traumatic cardiac arrest, it is unlikely that intubation carries the same life saving benefit as early defibrillation and bystander cardiopulmonary resuscitation (CPR). In trauma and paediatric patients, the current evidence base provides no imperative to extend the practice of prehospital intubation in urban systems. It would be ethical and pertinent to initiate a large, high quality randomised trial comparing the efficacy of competently practised emergency intubation with basic bag-valve-mask manoeuvres (BVM) in urban adult out-of-hospital non-traumatic cardiac arrest. PMID- 18425874 TI - WITHDRAWN: Acellular vaccines for preventing whooping cough in children. AB - BACKGROUND: Routine use of whole cell pertussis vaccines was suspended in some countries in the late 1970s and early 1980s, leading to a resurgence of whooping cough. Acellular pertussis vaccines containing purified or recombinant Bordetella pertussis antigens were developed in the hope that they would be as effective but less toxic than the whole cell vaccines. OBJECTIVES: The objective of this review was to assess the effects of acellular pertussis vaccines in children. SEARCH STRATEGY: The Cochrane Controlled Trials Register and Medline were searched up to January 1998. SELECTION CRITERIA: Double-blind randomised efficacy and safety trials of acellular pertussis vaccines in children, with active follow-up of participants and laboratory verification of pertussis cases. DATA COLLECTION AND ANALYSIS: One reviewer assessed trial quality and extracted data. MAIN RESULTS: Six efficacy trials and 45 safety trials were included. Acellular pertussis vaccines with three or more pertussis vaccines were more effective than those with one or two antigens. They were also more effective than one type of whole cell pertussis vaccine, but less effective than two other types of whole cell vaccines. Differences in trial design precluded pooling of the efficacy data and results should be interpreted with caution. Most systemic and local adverse events were significantly less common with acellular than with whole cell pertussis vaccines. AUTHORS' CONCLUSIONS: Multi-component acellular pertussis vaccines are effective, and show less adverse effects than whole cell pertussis vaccines. However in areas where whooping cough is more likely to be fatal, the higher toxicity of some whole cell vaccines may be offset by their increased effectiveness. PMID- 18425875 TI - Lumbar supports for prevention and treatment of low back pain. AB - BACKGROUND: Lumbar supports are used in the treatment of low-back pain patients, to prevent the onset of low-back pain (primary prevention) or to prevent recurrences of a low-back pain episode (secondary prevention). OBJECTIVES: To assess the effects of lumbar supports for prevention and treatment of non specific low-back pain. SEARCH STRATEGY: We updated the search in the Cochrane Central Register of Controlled Trials, PubMed, EMBASE, and CINAHL to December 2006. We also screened references given in relevant reviews and identified trials, and contacted experts to identify additional RCTs. SELECTION CRITERIA: Randomized controlled trials that reported on any type of lumbar supports as preventive or therapeutic intervention for non-specific low-back pain. DATA COLLECTION AND ANALYSIS: One review author generated the electronic search. Two review authors independently identified trials that met the inclusion criteria. One review author extracted data on the study population, interventions, and final results. The methodological quality and the clinical relevance were independently assessed by two review authors. Because it was not possible to perform a quantitative analysis, we performed a qualitative analysis in which the strength of evidence on the effectiveness of lumbar supports was classified as strong, moderate, limited, conflicting, or no evidence. MAIN RESULTS: Seven preventive studies (14,437 people) and eight treatment studies (1361 people) were included in this updated review. Overall, the methodological quality of the studies was rather low. Only five of the fifteen studies met 50% or more of the internal validity items. There was moderate evidence that lumbar supports are not more effective than no intervention or training in preventing low-back pain, and conflicting evidence whether lumbar supports are effective supplements to other preventive interventions. It is still unclear if lumbar supports are more effective than no or other interventions for the treatment of low-back pain. AUTHORS' CONCLUSIONS: There is moderate evidence that lumbar supports are not more effective than no intervention or training in preventing low-back pain, and conflicting evidence whether they are effective supplements to other preventive interventions. It remains unclear whether lumbar supports are more effective than no or other interventions for treating low-back pain. There is still a need for high quality randomized trials on the effectiveness of lumbar supports. One of the most essential issues to tackle in these future trials seems to be the realization of an adequate compliance. Special attention should be paid to different outcome measures, types of patients and types of lumbar support. PMID- 18425876 TI - Intermittent pneumatic compression for treating venous leg ulcers. AB - BACKGROUND: Intermittent pneumatic compression (IPC) is a mechanical method of delivering compression to swollen limbs that can be used to treat venous leg ulcers and limb swelling due to lymphoedema. This review analyses the evidence for the effectiveness of IPC as a treatment for venous leg ulcers. OBJECTIVES: To determine whether IPC increases the healing of venous leg ulcers. To determine the effects of IPC on health related quality of life of venous leg ulcer patients. SEARCH STRATEGY: We searched the Cochrane Wounds Group Specialised Register (December 2007); the Cochrane Central Register of Controlled Trials (CENTRAL) - The Cochrane Library Issue 4, 2007; Ovid MEDLINE - 2006 to November Week 2 2007; Ovid EMBASE - 2006 to 2007 Week 49 and Ovid CINAHL - 2006 to December Week 1 2007. SELECTION CRITERIA: Randomised controlled studies either comparing IPC with control (sham IPC or no IPC) or comparisons between IPC treatment regimens, in venous ulcer management were included. DATA COLLECTION AND ANALYSIS: Data extraction and assessment of study quality were undertaken by one author and checked by a second. MAIN RESULTS: Seven randomised controlled trials (including 367 people in total) were identified. Only one trial reported both allocation concealment and blinded outcome assessment. In one trial (80 people) more ulcers healed with IPC than with dressings (62% vs 28%; p=0.002). Four trials compared IPC with compression against compression alone. The first of these trials (45 people) found increased ulcer healing with IPC plus compression than with compression alone (relative risk for healing 11.4, 95% Confidence Interval 1.6 to 82). The remaining three trials (122 people) found no evidence of a benefit for IPC plus compression compared with compression alone. One small trial (16 people) found no difference between IPC (without additional compression) and compression bandages alone. One trial compared different ways of delivering IPC (104 people) and found that rapid IPC healed more ulcers than slow IPC (86% vs 61%; log rank p=0.003). AUTHORS' CONCLUSIONS: IPC may increase healing compared with no compression, but it is not clear whether it increases healing when added to treatment with bandages, or if it can be used instead of compression bandages. Rapid IPC was better than slow IPC in one trial. Further trials are required to determine whether IPC increases the healing of venous leg ulcers when used in modern practice where compression therapy is widely used. PMID- 18425877 TI - Information provision for stroke patients and their caregivers. AB - BACKGROUND: Research shows that stroke patients and their families are dissatisfied with the information provided and have a poor understanding of stroke and associated issues. OBJECTIVES: To assess the effectiveness of information provision strategies in improving the outcome for stroke patients and/or their identified caregivers. SEARCH STRATEGY: We searched the Cochrane Stroke Group Trials Register (last searched May 2007), the Cochrane Central Register of Controlled Trials (CENTRAL) (The Cochrane Library Issue 1, 2007), MEDLINE (1966 to March 2007), EMBASE (1980 to March 2007), CINAHL (1982 to March 2007), PsycINFO (1974 to March 2007), Science Citation Index and Social Science Citation Index (1981 to March 2007), Assia (1987 to March 2007), Index to UK theses (1970 to March 2007), Dissertation Abstracts (1961 to March 2007), ongoing trials and research registers, bibliographies of retrieved papers, relevant articles and books, and the Journal of Advanced Nursing. We also contacted researchers for additional information. SELECTION CRITERIA: Randomised trials involving patients or carers of patients with a clinical diagnosis of stroke or transient ischaemic attack (TIA) where an information intervention was compared with standard care, or where information and another therapy were compared with the other therapy alone. DATA COLLECTION AND ANALYSIS: Two review authors independently assessed trial eligibility and methodological quality and extracted data. Primary outcomes were knowledge about stroke and stroke services, and impact on mood. MAIN RESULTS: Seventeen trials involving 1773 patient and 1058 carer participants were included. Eight evaluated a passive and nine an active information intervention. Meta-analyses showed a significant effect in favour of the intervention on patient and carer knowledge, one aspect of patient satisfaction, and patient depression scores. There was no significant effect on number of cases of anxiety or depression in patients, carer mood or satisfaction, or death. Qualitative analyses found no strong evidence of an effect on other outcomes. Post-hoc subgroup analyses showed that active information had a significantly greater effect than passive information on patient mood but not on other outcomes. AUTHORS' CONCLUSIONS: There is evidence that information improves patient and carer knowledge of stroke, aspects of patient satisfaction, and reduces patient depression scores. However, the reduction in depression scores was small and probably clinically insignificant. Although the best way to provide information is still unclear there is some evidence that strategies that actively involve patients and carers and include planned follow up for clarification and reinforcement have a greater effect on patient mood. PMID- 18425878 TI - Delayed introduction of progressive enteral feeds to prevent necrotising enterocolitis in very low birth weight infants. AB - BACKGROUND: The introduction of progressive enteral feeds for very low birth weight infants is often delayed for several days or longer after birth due to concern that earlier introduction may not be tolerated and may increase the risk of necrotising enterocolitis. However, delaying enteral feeding could diminish the functional adaptation of the gastrointestinal tract and prolong the need for parenteral nutrition with its attendant infectious and metabolic risks. OBJECTIVES: To determine the effect of delayed introduction of progressive enteral feeds on the incidence of necrotising enterocolitis, mortality and other morbidities in very low birth weight infants. SEARCH STRATEGY: The standard search strategy of the Cochrane Neonatal Group was used. Searches were made of the Cochrane Central Register of Controlled Trials (CENTRAL, The Cochrane Library, Issue 4, 2007), MEDLINE (1966 - December 2007), EMBASE (1980 - December 2007), CINAHL (1982- December 2007), conference proceedings, and previous reviews. SELECTION CRITERIA: Randomised or quasi-randomised controlled trials that assessed the effect of delayed (after 96 hours' postnatal age) versus earlier introduction of progressive enteral feeds on the incidence of necrotising enterocolitis, mortality and other morbidities in very low birth weight infants. DATA COLLECTION AND ANALYSIS: The standard methods of the Cochrane Neonatal Group were used, with separate evaluation of trial quality and data extraction by two authors. Data were synthesised using a fixed effects model and reported using typical relative risk, typical risk difference and weighted mean difference. MAIN RESULTS: Two small trials in which a total of 74 infants participated were eligible for inclusion. Only a minority of participants were of extremely low birth weight or extreme preterm gestation. These trials provided no evidence that delayed introduction of progressive enteral feeds affected the incidence of necrotising enterocolitis, mortality or other neonatal morbidities. However, in view of the small number of participants, important beneficial or harmful effects cannot be excluded. AUTHORS' CONCLUSIONS: The available data are insufficient to inform clinical practice. Further large pragmatic randomised controlled trials are needed to determine how the timing of the introduction of progressive enteral feeds affects important clinical outcomes in very low birth weight infants, and particularly in extremely low birth weight or growth restricted infants. PMID- 18425879 TI - Bypass surgery for chronic lower limb ischaemia. AB - BACKGROUND: Surgical bypass of an occluded arterial segment is one of the mainstay treatments for patients with critical limb ischaemia (CLI). However, it was introduced without formal evaluation. OBJECTIVES: To determine the effects of bypass surgery in patients with CLI. SEARCH STRATEGY: The Cochrane Peripheral Vascular Diseases Group (PVD) searched their trials register (last searched November 2007) and the Cochrane Central Register of Controlled Trials (CENTRAL) in The Cochrane Library (last searched Issue 4, 2007). Principal trial investigators were also contacted. SELECTION CRITERIA: All randomised controlled trials (RCTs) of bypass surgery versus control or any other treatment. DATA COLLECTION AND ANALYSIS: For the update one author and PVD editorial staff extracted data and assessed trial quality. Unpublished data were obtained from trial investigators. Data were analyzed using Peto odds ratio (OR) or weighted mean difference (fixed and random effects models). MAIN RESULTS: Nineteen trials were identified. Eight involved a total of just over 1200 patients. Four trials compared bypass surgery with angioplasty (PTA) and one each with thromboendarterectomy, thrombolysis, exercise, and spinal cord stimulation. Four included patients with intermittent claudication (IC) and CLI, two were restricted to claudicants, and two to CLI. Vein grafts were used for distal reconstructions and synthetic prostheses for aorto-iliac or ilio-femoral bypasses. Six trials included mortality. In general, trial quality was good; blinding was not possible. Mortality and amputation rates did not differ significantly between bypass surgery and PTA; primary patency was significantly higher in the bypass group after 12 months (Peto OR 1.6, 95% CI 1.0 to 2.6) but not after four years (P = 0.14). In patients with lower CLI, surgery was associated with increased surgical complications (Peto OR 2.69, 95% CI 1.87 to 3.86) and longer hospital stays during the first year, mean stay 46.1 days (SD 53.9) compared with 36.4 days (SD 51.4) for those receiving PTA (P < 0.0001). Amputation rates were significantly lower in bypass compared with thrombolysis (Peto OR 0.2, 95% CI 0.1 to 0.6); mortality rates did not differ. Blood flow restoration was significantly greater in bypass than in thromboendarterectomy patients (Peto OR 9.2, 95% CI 1.7 to 50.6); mortality and amputation rates did not differ. Bypass surgery outcomes did not differ significantly from exercise or spinal cord stimulation. AUTHORS' CONCLUSIONS: There is limited evidence for the effectiveness of bypass surgery compared with other treatments; no studies compared bypass to no treatment. Further large trials are required. PMID- 18425880 TI - Buprenorphine maintenance versus placebo or methadone maintenance for opioid dependence. AB - BACKGROUND: Buprenorphine has been reported as an alternative to methadone for maintenance treatment of opioid dependence, but differing results are reported concerning its relative effectiveness indicating the need for an integrative review. OBJECTIVES: To evaluate the effects of buprenorphine maintenance against placebo and methadone maintenance in retaining patients in treatment and in suppressing illicit drug use. SEARCH STRATEGY: We searched the following databases up to October 2006: Cochrane Drugs and Alcohol Review Group Register, the Cochrane Controlled Trials Register, MEDLINE, EMBASE, Current Contents, Psychlit, CORK , Alcohol and Drug Council of Australia, Australian Drug Foundation, Centre for Education and Information on Drugs and Alcohol, Library of Congress databases, reference lists of identified studies and reviews, authors were asked about any other published or unpublished relevant RCT. SELECTION CRITERIA: Randomised clinical trials of buprenorphine maintenance versus placebo or methadone maintenance. DATA COLLECTION AND ANALYSIS: Authors separately and independently evaluated the papers and extracted data for meta-analysis. MAIN RESULTS: Twenty four studies met the inclusion criteria (4497 participants), all were randomised clinical trials, all but six were double-blind. The method of allocation concealment was not clearly described in the majority (20) of the studies, but where it was reported the methodological quality was good. Buprenorphine was statistically significantly superior to placebo medication in retention of patients in treatment at low doses (RR=1.50; 95% CI: 1.19 - 1.88), medium (RR=1.74; 95% CI: 1.06 - 2.87), and high doses (RR=1.74; 95% CI: 1.02 - 2.96). The high statistical heterogeneity prevented the calculation of a cumulative estimate. However, only medium and high dose buprenorphine suppressed heroin use significantly above placebo. Buprenorphine given in flexible doses was statistically significantly less effective than methadone in retaining patients in treatment (RR= 0.80; 95% CI: 0.68 - 0.95), but no different in suppression of opioid use for those who remained in treatment. Low dose methadone is more likely to retain patients than low dose buprenorphine (RR= 0.67; 95% CI: 0.52 - 0.87). Medium dose buprenorphine does not retain more patients than low dose methadone, but may suppress heroin use better. There was no advantage for medium dose buprenorphine over medium dose methadone in retention (RR=0.79; 95% CI:0.64 - 0.99) and medium dose buprenorphine was inferior in suppression of heroin use. AUTHORS' CONCLUSIONS: Buprenorphine is an effective intervention for use in the maintenance treatment of heroin dependence, but it is less effective than methadone delivered at adequate dosages. PMID- 18425881 TI - Intra-pleural fibrinolytic therapy versus conservative management in the treatment of adult parapneumonic effusions and empyema. AB - BACKGROUND: Pleural effusions and empyema may complicate lower respiratory tract infections. Treatment of these collections of pus includes surgical drainage and the use of intra-pleural fibrinolysis to break down fibrin bands that may cause loculation. OBJECTIVES: To conduct a systematic review of the benefit of adding intrapleural fibrinolytic therapy to intercostal tube drainage in the treatment of complicated para pneumonic effusions and empyema to reduce mortality or the need for subsequent surgical debridement of the pleural space. SEARCH STRATEGY: We searched the Cochrane Register of Controlled Trials (CENTRAL), MEDLINE and EMBASE. Trial authors were contacted for further information and details regarding the possibility of unpublished trials was requested. The most recent search was conducted in November 2006. SELECTION CRITERIA: All studies in the review were Randomised Controlled Trials in adult patients with post-pneumonic empyema or complicated parapneumonic effusions who had not had prior surgical intervention or trauma. The intervention was an intrapleural fibrinolytic agent (streptokinase or urokinase) via an intercostal chest drain (ICD) versus control, or a comparison of the two agents. DATA COLLECTION AND ANALYSIS: Two review authors independently extracted data . Study authors were contacted for further information. MAIN RESULTS: Seven studies met the eligibility criteria of the review, recruiting 761 participants. The only consistent end points in all trials were treatment failure, as gauged by the requirement for additional intervention including surgery or death. In studies where patients had either loculation and empyema, there was no significant difference in the risk of death with fibrinolytics (RR 1.08; 95% CI 0.69 to 1.68). When treatment failure was considered as surgical intervention, fibrinolytics reduced the risk of this outcome (RR 0.63; 95% CI 0.46 to 0.85), but there is discordance between earlier positive studies and the more recent negative study by Maskell. AUTHORS' CONCLUSIONS: Intrapleural fibrinolytic therapy confers significant benefit in reducing the requirement for surgical intervention for patients in the early studies included in this review but not in the more recently published Maskell study. The reasons for this difference are uncertain. Separate subgroup analysis of proven loculated/septated effusions from the available data in our meta analysis suggests a potential overall treatment benefit with fibrinolytics, but these results should be treated with caution as the data are incomplete and the benefit is not significant in the subgroup of high quality trials (Cochrane Grade A). Intrapleural fibrinolytics have not been shown to significantly increase adverse events, but the confidence interval is too wide to firmly exclude this possibility. PMID- 18425882 TI - Continuous heparin infusion to prevent thrombosis and catheter occlusion in neonates with peripherally placed percutaneous central venous catheters. AB - BACKGROUND: Complications associated with peripherally placed percutaneous central venous catheters (PCVC) in neonates include mechanical complications (catheter thrombosis, occlusion or dislodgement) and infection. Strategies to prevent catheter thrombosis and occlusion include the use of heparin. However, heparin is known to be associated with complications such as bleeding and thrombocytopenia. OBJECTIVES: PRIMARY OBJECTIVE: To assess the effectiveness of heparin for prevention of catheter related thrombosis. SECONDARY OBJECTIVES: To assess the effectiveness of heparin on catheter occlusion, duration of catheter patency, catheter related sepsis and complications associated with the use of heparin. SEARCH STRATEGY: A literature search of MEDLINE, EMBASE, CINAHL from their inception to December 2007, The Cochrane Library (Issue 4, 2007) and abstracts from the annual meetings of the Pediatric Academic Societies was performed without language restrictions. SELECTION CRITERIA: Randomized or quasi randomized clinical trials of neonates where heparin infusion was compared to placebo or no treatment for prevention of any of the complications related to peripherally placed PCVC were included. DATA COLLECTION AND ANALYSIS: The methodological quality of included trials was assessed using criteria for masking of randomization, masking of intervention, completeness of follow-up and masking of outcome measurement. Data on relevant outcomes were extracted and the effect size was estimated by calculating relative risk (RR), risk difference (RD) and associated 95% confidence intervals (CI). MAIN RESULTS: Three randomized trials were identified. Two trials of adequate methodology met the eligibility criteria. These studies included 267 neonates. There was reduced risk of catheter occlusion (typical RR 0.28, 95% CI 0.15, 0.53, NNT 5, 95% CI 3, 8). There was no statistically significant difference in the duration of catheter patency when analyzed as continuous data; however, in one study survival analyses identified benefit with heparin (adjusted hazard ratio 0.55, 95% CI 0.36, 83); (Shah 2007). This could be due to higher incidence of elective removal of catheters in neonates at the completion of therapy in the heparin group (63% vs. 42%; p = 0.002) (Shah 2007). There was no statistically significant differences in the risk of thrombosis (typical RR 0.93, 95% CI 0.58, 1.51), catheter related sepsis (typical RR 1.96, 95% CI 0.50, 7.60), or extension of intraventricular hemorrhage (typical RR 0.87, 95% CI 0.25, 3.03) between the two groups. AUTHORS' CONCLUSIONS: IMPLICATIONS FOR PRACTICE: Prophylactic use of heparin for peripherally placed PCVC allows a greater number of infants to complete their intended use (complete therapy) by reducing occlusion. Evidence from this systematic review support the prophylactic use of heparin for PCVC in neonates at a dose of 0.5 IU/kg/hr. IMPLICATIONS FOR RESEARCH: None of these studies was powered to evaluate a lower incidence rate of adverse events. If this therapy is adopted in routine practice, monitoring of side effects is indicated. PMID- 18425883 TI - Rocuronium versus succinylcholine for rapid sequence induction intubation. AB - BACKGROUND: Patients requiring emergency endotracheal intubation often require a rapid sequence induction (RSI) intubation technique to protect against aspiration or increased intracranial pressure, or to facilitate intubation. Succinylcholine is the most commonly used muscle relaxant because of its fast onset and short duration; unfortunately, it can have serious side effects. Rocuronium has been suggested as an alternative to succinylcholine for intubation. This meta-analysis is an update since our initial Cochrane systematic review in 2003. OBJECTIVES: To determine if rocuronium creates comparable intubating conditions to succinylcholine during RSI intubation. Comparisons were made based on dose of rocuronium, narcotic use, emergency versus elective intubation, age and induction agent. The primary outcome was excellent intubation conditions. The secondary outcome was acceptable conditions. SEARCH STRATEGY: In our initial systematic review we searched all databases until March 2000. We have updated that search and searched the Cochrane Central Register of Controlled Trials (The Cochrane Library, 2007 issue 3), MEDLINE (1966 to June Week 3 2007), EMBASE (1988 to 2007 Week 26) for randomized controlled trials or controlled clinical trials relating to the use of rocuronium and succinylcholine. We included foreign language journals and handsearched the references of identified studies for additional citations. SELECTION CRITERIA: We included all trials meeting the inclusion criteria (comparison of rocuronium and succinylcholine, main outcomes of intubation conditions). DATA COLLECTION AND ANALYSIS: Two authors (JP, JL or VS) independently extracted data and assessed methodological quality for allocation concealment. We combined the outcomes in RevMan using relative risk (RR) with a random-effects model. MAIN RESULTS: In our initial systematic review we identified 40 studies and included 26. In this update we identified a further 18 studies and included 11. In total, we identified 58 potential studies; 37 were combined for meta-analysis. Overall, succinylcholine was superior to rocuronium, RR 0.86 (95% confidence interval (95% CI) 0.80 to 0.92) (n = 2690). In the group that used propofol for induction, the intubation conditions were superior with succinylcholine (RR 0.88, 95% CI 0.80 to 0.97) (n = 1183). This is contrary to our previous meta-analysis results where we reported that intubation conditions were superior in the rocuronium group when propofol was used. We found no statistical difference in intubation conditions when succinylcholine was compared to 1.2mg/kg rocuronium; however, succinylcholine was clinically superior as it has a shorter duration of action. AUTHORS' CONCLUSIONS: Succinylcholine created superior intubation conditions to rocuronium when comparing both excellent and clinically acceptable intubating conditions. PMID- 18425884 TI - Techniques for surgical retrieval of sperm prior to intra-cytoplasmic sperm injection (ICSI) for azoospermia. AB - BACKGROUND: Azoospermia, the absence of sperm in ejaculated semen, is the most severe form of male-factor infertility and is present in approximately 5% of all investigated infertile couples. The advent of intra-cytoplasmic sperm injection (ICSI) has transformed treatment of this type of severe male-factor infertility. Sperm can be retrieved for ICSI from either the epididymis or the testis, depending on the type of azoospermia. OBJECTIVES: To evaluate the efficacy of the various surgical retrieval techniques for men with obstructive or non-obstructive azoospermia prior to ICSI. SEARCH STRATEGY: We searched the Cochrane Menstrual Disorders and Subfertility Group Trials Register (November 2007), Cochrane Central Register of Controlled Trials (CENTRAL) (The Cochrane Library 2007, Issue 4), MEDLINE (1966 to November 2007), EMBASE (1980 to November 2007), Biological Abstracts (1980 to November 2007), and reference lists of identified articles. SELECTION CRITERIA: Randomised controlled trials (RCTs) comparing the effectiveness of different sperm-retrieval techniques in men with azoospermia prior to ICSI. Due to the lack of RCTs, non-randomised trials that used the participants as their own control were also considered in the review but their results were not included in the meta-analysis. DATA COLLECTION AND ANALYSIS: Two review authors independently assessed trial quality and extracted data. Study authors were contacted for additional information. MAIN RESULTS: The search was revised and re-run in November 2007. No new trials were located therefore the results of the updated review remain unchanged from those published in 2006. Two trials involving 98 men were included. The first small RCT had 59 participants and compared two epididymal techniques. The trial gave limited evidence that microsurgical epididymal sperm aspiration (MESA) achieved a significantly lower pregnancy rate (one pregnancy in 29 procedures compared with seven pregnancies in 30 procedures; OR 0.19, 95% CI 0.04 to 0.83) and fertilisation rate (OR 0.16, 95% CI 0.05 to 0.48) than the micropuncture with perivascular nerve stimulation technique. The other RCT comparing two testicular aspiration techniques (TSA) in 39 participants gave no statistically significant evidence for the superiority of the ultrasound-guided technique compared to the aspiration technique without ultrasound. TSA with ultrasound resulted in pregnancy in three out of 16 participants compared with four out of 23 participants (OR 1.10, 95% CI 0.21 to 5.74). AUTHORS' CONCLUSIONS: There is insufficient evidence to recommend any specific sperm retrieval technique for azoospermic men undergoing ICSI. In the absence of evidence to support more invasive or more technically difficult methods, the review authors recommend the least invasive and simplest technique available. Further randomised trials are warranted, preferably multi-centred trials. The classification of azoospermia as obstructive and non-obstructive appears to be relevant to a successful clinical outcome and a distinction according to the cause of azoospermia is important for future clinical trials. PMID- 18425885 TI - Mistletoe therapy in oncology. AB - BACKGROUND: Mistletoe extracts are commonly used in cancer patients. It is claimed that they improve survival and quality of life (QOL) in cancer patients. OBJECTIVES: To determine the effectiveness, tolerability and safety of mistletoe extracts given either as monotherapy or adjunct therapy for patients with cancer. SEARCH STRATEGY: Search sources included the Cochrane Central Register of Controlled Trials (CENTRAL, Issue 3, 2007) Cochrane Complementary Medicine Field Registry of randomized clinical trials (RCTs) and controlled clinical trials, MEDLINE, EMBASE, HEALTHSTAR, INT. HEALTH TECHNOLOGY ASSESSMENT, SOMED, AMED, BIOETHICSLINE, BIOSIS, CancerLit, CATLINE, CISCOM (August 2007). For the search the Standard Operating Procedures of the Information System in Health Economics at the German Institute for Medical Documentation and Information (DIMDI) were utilized. Reference lists of relevant articles and authors extensive files were searched for additional studies. Manufacturers of mistletoe preparations were contacted. SELECTION CRITERIA: We included RCTs of adults with cancer of any type. The interventions were mistletoe extracts as sole treatments or given concomitantly with chemo- or radiotherapy. The outcome measures were survival times, tumor response, QOL, psychological distress, adverse effects from antineoplastic treatment and safety of mistletoe extracts. DATA COLLECTION AND ANALYSIS: Three review authors independently assessed trials for inclusion in the review. All review authors independently took part in the extraction of data and assessment of study quality and clinical relevance. Disagreements were resolved by consensus. Study authors were contacted where information was unclear. Methodological quality was narratively described and additionally assessed with the Delphi list and the Jadad score. High methodological quality was defined if six out of nine Delphi criteria, or four out of five Jadad criteria were fulfilled. Results were presented qualitatively. MAIN RESULTS: Eighty studies were identified. Fifty-eight were excluded for various reasons, usually as there was no prospective trial design with randomised treatment allocation. Of the 21 included studies 13 provided data on survival, 7 on tumour response, 16 on measures of QOL or psychological outcomes, or prevalence of chemotherapy-related adverse effects and 12 on side effects of mistletoe treatment; overall comprising 3484 randomised cancer patients. Interventions evaluated were 5 preparations of mistletoe extracts from 5 manufacturers and one commercially not available preparation. The general reporting of RCTs was poor. Of the 13 trials investigating survival, 6 showed some evidence of a benefit, but none of them was of high methodological quality. The results of two trials in patients with melanoma and head and neck cancer gave some evidence that the used mistletoe extracts are not effective for improving survival. Of the 16 trials investigating the efficacy of mistletoe extracts for either improving QOL, psychological measures, performance index, symptom scales or the reduction of adverse effects of chemotherapy, 14 showed some evidence of a benefit, but only 2 of them including breast cancer patients during chemotherapy were of higher methodological quality. Data on side effects indicated that, depending on the dose, mistletoe extracts were usually well tolerated and had few side effects. AUTHORS' CONCLUSIONS: The evidence from RCTs to support the view that the application of mistletoe extracts has impact on survival or leads to an improved ability to fight cancer or to withstand anticancer treatments is weak. Nevertheless, there is some evidence that mistletoe extracts may offer benefits on measures of QOL during chemotherapy for breast cancer, but these results need replication. Overall, more high quality, independent clinical research is needed to truly assess the safety and effectiveness of mistletoe extracts. Patients receiving mistletoe therapy should be encouraged to take part in future trails. PMID- 18425886 TI - Long-term results of laparoscopic colorectal cancer resection. AB - BACKGROUND: Although minimally invasive surgery has been accepted for a variety of disorders, laparoscopic resection of colorectal cancer is performed by few. Concern about oncological radicality and long term outcome has limited the adoption of laparoscopic surgery for colorectal cancer. OBJECTIVES: To determine long-term outcome after laparoscopically-assisted versus open surgery for non metastasised colorectal cancer. SEARCH STRATEGY: The Cochrane library, EMBASE, Pub med and Cancer Lit were searched for published and unpublished randomised controlled trials. SELECTION CRITERIA: Randomised clinical trials comparing laparoscopically-assisted and open surgery for non-metastasised colorectal cancer were included. Studies that did not report any long-term outcomes were excluded. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed the studies and extracted data. RevMan 4.2 was used for statistical analysis. MAIN RESULTS: Thirty-three randomised clinical trials (RCT) comparing laparoscopically-assisted versus open surgery for colorectal cancer were identified. Twelve of these trials, involving 3346 patients, reported long-term outcome and were included in the current analysis. No significant differences in the occurrence of incisional hernia, reoperations for incisional hernia or reoperations for adhesions were found between laparoscopically assisted and open surgery (2 RCT, 474 pts, 7.9% vs 10.9%;P = 0.32 and 2 RCT, 474 pts, 4.0% vs 2.8%; P = 0.42 and 1 RCT, 391 pts, 1.1% vs 2.5%;P = 0.30, respectively). Rates of recurrence at the site of the primary tumor were similar (colon cancer: 4 RCT, 938 pts, 5.2% vs 5.6%; OR (fixed) 0.84 (95% CI 0.47 to 1.52)(P = 0.57); rectal cancer: 4 RCT, 714 pts, 7.2% vs 7.7%; OR (fixed) 0.81 (95% CI 0.45 to 1.43) (P = 0.46). No differences in the occurrence of port-site/wound recurrences were observed (P=0.16). Similar cancer related mortality was found after laparoscopic surgery compared to open surgery ( colon cancer: 5 RCT, 1575 pts, 14.6% vs 16.4%; OR (fixed) 0.80 (95% CI 0.61 to 1.06) (P=0.15); rectal cancer: 3 RCT, 578 pts, 9.2% vs 10.0%; OR (fixed) 0.66 (95% CI 0.37 to 1.19) (P=0.16). Four studies were included in the meta-analyses on hazard ratios for tumour recurrence in laparoscopic colorectal cancer surgery. No significant difference in recurrence rate was observed between laparoscopic and open surgery (hazard ratio for tumour recurrence in the laparoscopic group 0.92; 95% CI 0.76-1.13). No significant difference in tumour recurrence between laparoscopic and open surgery for colon cancer was observed (hazard ratio for tumour recurrence in the laparoscopic group 0.86; 95% CI 0.70-1.08). AUTHORS' CONCLUSIONS: Laparoscopic resection of carcinoma of the colon is associated with a long term outcome no different from that of open colectomy. Further studies are required to determine whether the incidence of incisional hernias and adhesions is affected by method of approach. Laparoscopic surgery for cancer of the upper rectum is feasible, but more randomised trials need to be conducted to assess long term outcome. PMID- 18425887 TI - WITHDRAWN: Amino acids for amyotrophic lateral sclerosis / motor neuron disease. AB - BACKGROUND: Amyotrophic lateral sclerosis, also known as motor neuron disease, is a progressive neuromuscular disease that causes disability and eventual death. Various amino acid preparations, the three branched-chain amino acids (L-leucine, L-valine and L-isoleucine) or, alternatively, L-threonine have been used as experimental therapy. OBJECTIVES: To examine the efficacy of amino acid therapies in prolonging survival and/or slowing the progression of amyotrophic lateral sclerosis/motor neuron disease. SEARCH STRATEGY: We searched the Cochrane Neuromuscular Disease Group trials register (searched February 2003), MEDLINE (from January 1966 to December 2002) and EMBASE (from January 1980 to December 2002) databases and reports of specialist conferences. Authors of known studies were contacted. SELECTION CRITERIA: We included randomised or quasi-randomised trials of participants with a clinical diagnosis of amyotrophic lateral sclerosis/motor neuron disease treated with all combinations of amino acids. Our primary outcome measure was survival determined by a pooled hazard ratio of all studies. Our secondary outcome measures were (in order of priority): survival at six and 12 months, muscle strength, any validated rating scale of physical function, quality of life, proportion of patients completing therapy and proportion of patients reporting adverse events attributable to treatment. DATA COLLECTION AND ANALYSIS: We identified six eligible trials and rejected a further seven because of incomplete data or inadequate duration. Eligible studies were rated for methodological quality and missing data sought from the authors. After this examination two studies were excluded from analysis. Our pooled survival analysis was performed by the Parmar method, other statistical calculations were done using the Review Manager 4.2 software package. MAIN RESULTS: No benefit could be demonstrated for either branched-chain amino acids or L-threonine in improving survival in amyotrophic lateral sclerosis/motor neuron disease. Neither could we find evidence of an effect of either treatment on muscle strength or disability as measured by functional rating scales. No study assessed quality of life. Both branched-chain amino acids and L-threonine appeared well tolerated and caused a degree of adverse events comparable to that of the control medication. AUTHORS' CONCLUSIONS: There is no evidence to support a beneficial effect of either branched-chain amino acids or L-threonine in amyotrophic lateral sclerosis/motor neuron disease. PMID- 18425888 TI - Pharmacological and nutritional treatment for McArdle disease (Glycogen Storage Disease type V). AB - BACKGROUND: McArdle disease (Glycogen Storage Disease type V) is caused by the absence of the glycolytic enzyme, muscle phosphorylase. People present with exercise-induced pain, cramps, fatigue, and myoglobinuria, which can result in acute renal failure if it is severe. OBJECTIVES: To systematically review the evidence from randomised controlled trials of pharmacological or nutritional treatments in improving exercise performance and quality of life in McArdle disease. SEARCH STRATEGY: We updated the review by searching the Cochrane Neuromuscular Disease Group Trials Register (November 2007), MEDLINE (January 1966 to November 2007) and EMBASE (January 1980 to November 2007) using the search terms 'McArdle disease' and its synonym 'Glycogen Storage Disease type V'. SELECTION CRITERIA: We included randomised controlled trials (including crossover studies) and quasi-randomised trials. Open trials and individual patient studies with no participant or observer blinding were included in the discussion. Types of interventions included any pharmacological agent or micronutrient or macronutrient supplementation. Primary outcome measures included any objective assessment of exercise endurance (for example aerobic capacity (VO(2)) max, walking speed, muscle force or power and improvement in fatiguability). Secondary outcome measures included metabolic changes (such as reduced plasma creatine kinase activity and a reduction in the frequency of myoglobinuria), subjective measures (including quality of life scores and indices of disability) and serious adverse events. DATA COLLECTION AND ANALYSIS: Three review authors checked the titles and abstracts identified by the search and reviewed the manuscripts. Two review authors (RQ and RB) independently assessed methodological quality of the full text of potentially relevant studies and extracted data onto a specially designed form. MAIN RESULTS: We reviewed 24 studies. Twelve trials fulfilled the criteria for inclusion, with two being first identified in this update. The 12 excluded trials are included in the discussion. The largest treatment trial included 19 cases. The other trials included fewer than 12 cases. As there were only single trials for a given intervention we were unable to undertake a meta analysis. AUTHORS' CONCLUSIONS: There is no evidence of significant benefit from any specific nutritional or pharmacological treatment in McArdle disease. In one small trial low dose creatine produced slight benefit but high dose creatine caused myalgia. Ingestion of oral sucrose immediately before exercise reduced perceived ratings of exertion and heart rate and improved exercise tolerance. This treatment will not influence sustained or unexpected exercise and may cause significant weight gain. A carbohydrate rich diet did benefit patients. Because of the rarity of McArdle disease, there is a need to develop international multicentre collaboration and standardised assessment protocols for future treatment trials. PMID- 18425889 TI - Heparin versus placebo for acute coronary syndromes. AB - BACKGROUND: Acute coronary syndromes (ACS) represent a spectrum of disease including unstable angina (UA) and non-ST segment myocardial infarction (NSTEMI). Despite treatment with aspirin, beta-blockers and nitroglycerin, UA/NSTEMI is still associated with significant morbidity and mortality. Although emerging evidence suggests that low molecular weight heparin (LMWH) is more efficacious compared to unfractionated heparin (UFH), there is limited data to support the role of heparins as a drug class in the treatment of ACS. OBJECTIVES: To determine the effect of heparins (UFH and LMWH) compared with placebo for the treatment of patients with ACS. SEARCH STRATEGY: We searched the Cochrane Central Register of Controlled Trials on The Cochrane Library (issue 4, 2002), MEDLINE (1966 to May 2002), EMBASE (1980 to May 2002) and CINAHL (1982 to May 2002). Authors of included studies and pharmaceutical industry representatives were contacted to determine if unpublished studies which met the inclusion criteria were available. SELECTION CRITERIA: Randomized controlled trials of parenteral UFH or LMWH versus placebo in people with ACS (UA or NSTEMI). DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed quality of studies. Data were extracted independently by two reviewers. Study authors were contacted to verify and clarify missing data. MAIN RESULTS: Eight studies (3118 participants) were included in this review. We found no evidence for difference in overall mortality between the groups treated with heparin and placebo (RR = 0.84, 95% CI 0.36 to 1.98). Heparins reduced the occurrence of MI (RR = 0.40, 95% CI 0.25 to 0.63, NNT = 33). An increase in the incidence of minor bleeds (RR = 6.80, 95% CI 1.23 to 37.49, NNH = 17). AUTHORS' CONCLUSIONS: Compared to placebo, patients treated with heparins had similar risk of mortality, revascularization, recurrent angina, major bleeding and thrombocytopenia. However, those treated with heparins had decreased risk of MI and a higher incidence of minor bleeding. PMID- 18425890 TI - Gluten- and casein-free diets for autistic spectrum disorder. AB - BACKGROUND: It has been suggested that peptides from gluten and casein may have a role in the origins of autism and that the physiology and psychology of autism might be explained by excessive opioid activity linked to these peptides. Research has reported abnormal levels of peptides in the urine and cerebrospinal fluid of people with autism. OBJECTIVES: To determine the efficacy of gluten and/or casein free diets as an intervention to improve behaviour, cognitive and social functioning in individuals with autism. SEARCH STRATEGY: The following electronic databases were searched: CENTRAL(The Cochrane Library Issue 2, 2007), MEDLINE (1966 to April 2007), PsycINFO (1971 to April 2007), EMBASE (1974 to April 2007), CINAHL (1982 to April 2007), ERIC (1965 to 2007), LILACS (1982 to April 2007), and the National Research register 2007 (Issue1). Review bibliographies were also examined to identify potential trials. SELECTION CRITERIA: All randomised controlled trials (RCT) involving programmes which eliminated gluten, casein or both gluten and casein from the diets of individuals diagnosed with an autistic spectrum disorder. DATA COLLECTION AND ANALYSIS: Abstracts of studies identified in searches of electronic databases were assessed to determine inclusion by two independent authors The included trials did not share common outcome measures and therefore no meta-analysis was possible. Data are presented in narrative form. MAIN RESULTS: Two small RCTs were identified (n = 35). No meta-analysis was possible. There were only three significant treatment effects in favour of the diet intervention: overall autistic traits, mean difference (MD) = -5.60 (95% CI -9.02 to -2.18), z = 3.21, p=0.001 (Knivsberg 2002) ; social isolation, MD = -3.20 (95% CI -5.20 to 1.20), z = 3.14, p = 0.002) and overall ability to communicate and interact, MD = 1.70 (95% CI 0.50 to 2.90), z = 2.77, p = 0.006) (Knivsberg 2003). In addition three outcomes showed no significant difference between the treatment and control group and we were unable to calculate mean differences for ten outcomes because the data were skewed. No outcomes were reported for disbenefits including harms. AUTHORS' CONCLUSIONS: Research has shown of high rates of use of complementary and alternative therapies (CAM) for children with autism including gluten and/or casein exclusion diets. Current evidence for efficacy of these diets is poor. Large scale, good quality randomised controlled trials are needed. PMID- 18425891 TI - Progestogen for preventing miscarriage. AB - BACKGROUND: Progesterone, a female sex hormone, is known to induce secretory changes in the lining of the uterus essential for successful implantation of a fertilised egg. It has been suggested that a causative factor in many cases of miscarriage may be inadequate secretion of progesterone. Therefore, progestogens have been used, beginning in the first trimester of pregnancy, in an attempt to prevent spontaneous miscarriage. OBJECTIVES: To determine the efficacy and safety of progestogens as a preventative therapy against miscarriage. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group's Trials Register (January 2008), CENTRAL (The Cochrane Library 2006, Issue 4), MEDLINE (1966 to June 2006), EMBASE (1980 to June 2006), CINAHL (1982 to June 2006), NHMRC Clinical Trials Register (June 2006) and Meta-Register (June 2006). We searched references from relevant articles, attempting to contact authors where necessary, and contacted experts in the field for unpublished works. SELECTION CRITERIA: Randomised or quasi-randomized controlled trials comparing progestogens with placebo or no treatment given in an effort to prevent miscarriage. DATA COLLECTION AND ANALYSIS: Two review authors assessed trial quality and extracted data. MAIN RESULTS: Fifteen trials (2118 women) are included. The meta-analysis of all women, regardless of gravidity and number of previous miscarriages, showed no statistically significant difference in the risk of miscarriage between progestogen and placebo or no treatment groups (Peto odds ratio (Peto OR) 0.98; 95% confidence interval (CI) 0.78 to 1.24) and no statistically significant difference in the incidence of adverse effect in either mother or baby. In a subgroup analysis of three trials involving women who had recurrent miscarriages (three or more consecutive miscarriages), progestogen treatment showed a statistically significant decrease in miscarriage rate compared to placebo or no treatment (Peto OR 0.38; 95% CI 0.20 to 0.70). No statistically significant differences were found between the route of administration of progestogen (oral, intramuscular, vaginal) versus placebo or no treatment. AUTHORS' CONCLUSIONS: There is no evidence to support the routine use of progestogen to prevent miscarriage in early to mid-pregnancy. However, there seems to be evidence of benefit in women with a history of recurrent miscarriage. Treatment for these women may be warranted given the reduced rates of miscarriage in the treatment group and the finding of no statistically significant difference between treatment and control groups in rates of adverse effects suffered by either mother or baby in the available evidence. Larger trials are currently underway to inform treatment for this group of women. PMID- 18425892 TI - Interventions for treating collagenous colitis. AB - BACKGROUND: Collagenous colitis is a cause of chronic diarrhea. Treatment has been based mainly on anecdotal evidence. This review was performed to identify therapies for collagenous colitis that have been proven in randomized trials. OBJECTIVES: To determine effective treatments for patients with collagenous colitis. SEARCH STRATEGY: Relevant papers published between 1970 and December 2007 were identified via the MEDLINE and PUBMED databases. Manual searches from the references of identified papers, as well as review papers on collagenous or microscopic colitis were performed to identify additional studies. Abstracts from major gastroenterological meetings were searched to identify research submitted in abstract form only. Finally, the Cochrane Controlled Trials Register and the Cochrane Inflammatory Bowel Disease and Functional Bowel Disorders Group Specialized Trials Register were searched for other studies. SELECTION CRITERIA: Ten randomized trials were identified. Seven of these compared active treatment to placebo for treating active disease. Of these, 1 trial studied bismuth subsalicylate, 1 trial studied Boswellia serrata extract, 3 trials studies budesonide, 1 trial studied prednisolone, and 1 trial studied probiotics. One trial compared mesalamine to mesalamine + cholestyramine for treating active disease. Two trials compared budesonide to placebo in maintaining response induced by budesonide. DATA COLLECTION AND ANALYSIS: Data were extracted independently by each author onto 2x2 tables (treatment versus comparator and response versus no response). For therapies assessed in one trial only, P-values were derived using the chi-square test. For therapies assessed in more than one trial, summary test statistics were derived using the Peto odds ratio and 95% confidence intervals. Data were combined for analysis only if the outcomes were sufficiently similar in definition. MAIN RESULTS: In treating active disease, there were 9 patients with collagenous colitis in the trial studying bismuth subsalicylate (nine 262 mg tablets daily for 8 weeks). Clinical response occurred in 100% of patients who received bismuth subsalicylate compared to 0% of patients who received placebo (P = 0.03). Thirty-one patients were enrolled in the trial studying Boswellia serrata extract (three 400 mg capsules daily for 8 weeks). Clinical response occurred in 44% of patients who received Boswellia serrata extract compared to 27% of patients who received placebo (P = 0.32). A total of 94 patients were enrolled in 3 trials studying budesonide (9 mg daily or in a tapering schedule for 6 to 8 weeks). Clinical response occurred in 81% of patients who received budesonide compared to 17% of patients who received placebo (P < 0.00001). The pooled odds ratio for clinical response to treatment with budesonide was 12.32 (95% CI 5.53 to 27.46), with a number needed to treat of 2 patients. Statistically significant histological response occurred with treatment in all 3 trials studying budesonide therapy. Eleven patients were enrolled in the trial studying prednisolone (50 mg daily for 2 weeks). Clinical response occurred in 63% of patients who received prednisolone compared to 0% who received placebo (P = 0.15). Twenty-nine patients were enrolled in the trial studying probiotics (2 capsules containing 0.5 x 10(10) CFU each of L. acidophilus LA-5 and B. animalis subsp. lactis strain BB-12 twice daily for 12 weeks). Clinical response occurred in 29% of patients who received probiotics compared to 13% of patients who received placebo (P = 0.38). Twenty-three patients were enrolled in the trial studying mesalamine (800 mg three times daily) with or without cholestyramine (4 g daily) for 6 months. Clinical response occurred in 73% of patients who received mesalamine alone compared to 100% of patients who received mesalamine + cholestyramine (P = 0.14). In maintaining response, 80 patients who had responded to open-label budesonide were enrolled in 2 trials studying budesonide (6 mg daily for 6 months). Clinical response was maintained in 83% of patients who received budesonide compared to 28% of patients who received placebo (P = 0.0002). The pooled odds ratio for maintenance of clinical response to treatment with budesonide was 8.40 (95% CI 2.73 to 25.81), with a number needed to treat of 2 patients. Histological response was maintained in 48% of patients who received budesonide compared to 15% of patients who received placebo (P = 0.002). AUTHORS' CONCLUSIONS: Budesonide is effective for inducing and maintaining clinical and histological response in patients with collagenous colitis. The evidence for benefit with bismuth subsalicylate and for mesalamine with or without cholestyramine is weak. There is no evidence for the effectiveness of Boswellia serrata extract, prednisolone, or probiotics. These agents and other therapies require further study. PMID- 18425893 TI - Chemoprophylaxis and intermittent treatment for preventing malaria in children. AB - BACKGROUND: Malaria causes repeated illness in children living in endemic areas. Policies of giving antimalarial drugs at regular intervals (prophylaxis or intermittent treatment) are being considered for preschool children. OBJECTIVES: To evaluate prophylaxis and intermittent treatment with antimalarial drugs to prevent malaria in young children living in malaria-endemic areas. SEARCH STRATEGY: We searched the Cochrane Infectious Diseases Group Specialized Register (August 2007), CENTRAL (The Cochrane Library 2007, Issue 3), MEDLINE (1966 to August 2007), EMBASE (1974 to August 2007), LILACS (1982 to August 2007), mRCT (February 2007), and reference lists of identified trials. We also contacted researchers. SELECTION CRITERIA: Individually randomized and cluster-randomized controlled trials comparing antimalarial drugs given at regular intervals (prophylaxis or intermittent treatment) with placebo or no drug in children aged one month to six years or less living in a malaria-endemic area. DATA COLLECTION AND ANALYSIS: Two authors independently extracted data and assessed methodological quality. We used relative risk (RR) or weighted mean difference with 95% confidence intervals (CI) for meta-analyses. Where we detected heterogeneity and considered it appropriate to combine the trials, we used the random-effects model (REM). MAIN RESULTS: Twenty-one trials (19,394 participants), including six cluster-randomized trials, met the inclusion criteria. Prophylaxis or intermittent treatment with antimalarial drugs resulted in fewer clinical malaria episodes (RR 0.53, 95% CI 0.38 to 0.74, REM; 7037 participants, 10 trials), less severe anaemia (RR 0.70, 95% CI 0.52 to 0.94, REM; 5445 participants, 9 trials), and fewer hospital admissions for any cause (RR 0.64, 95% CI 0.49 to 0.82; 3722 participants, 5 trials). We did not detect a difference in the number of deaths from any cause (RR 0.90, 95% CI 0.65 to 1.23; 7369 participants, 10 trials), but the CI do not exclude a potentially important difference. One trial reported three serious adverse events with no statistically significant difference between study groups (1070 participants). Eight trials measured morbidity and mortality six months to two years after stopping regular antimalarial drugs; overall, there was no statistically significant difference, but participant numbers were small. AUTHORS' CONCLUSIONS: Prophylaxis and intermittent treatment with antimalarial drugs reduce clinical malaria and severe anaemia in preschool children. PMID- 18425894 TI - Antiviral medications for preventing cytomegalovirus disease in solid organ transplant recipients. AB - BACKGROUND: The risk of cytomegalovirus (CMV) infection in solid organ transplant recipients has resulted in the frequent use of prophylaxis with the aim of preventing the clinical syndrome associated with CMV infection. OBJECTIVES: To determine the benefits and harms of antiviral medications to prevent CMV disease and all-cause mortality in solid organ transplant recipients. SEARCH STRATEGY: We searched the Cochrane Central Register of Controlled Trials, MEDLINE, EMBASE, reference lists and abstracts from conference proceedings without language restriction. Date of last search: February 2007 SELECTION CRITERIA: Randomised controlled trials (RCTs) and quasi-RCTs comparing antiviral medications with placebo or no treatment, comparing different antiviral medications and comparing different regimens of the same antiviral medications in recipients of any solid organ transplant. DATA COLLECTION AND ANALYSIS: Statistical analyses were performed using the random effects model and results expressed as relative risk (RR) for dichotomous outcomes with 95% confidence intervals (CI). Subgroup analysis and univariate meta-regression were performed using restricted maximum likelihood to estimate the between study variance. Multivariate meta-regression was performed to investigate whether the results were altered after allowing for differences in drugs used, organ transplanted and recipient CMV serostatus at the time of transplantation. MAIN RESULTS: Thirty four studies (3850 participants) were identified. Prophylaxis with aciclovir, ganciclovir or valaciclovir compared with placebo or no treatment significantly reduced the risk for CMV disease (19 studies; RR 0.42, 95% CI 0.34 to 0.52), CMV infection (17 studies; RR 0.61, 95% CI 0.48 to 0.77), and all-cause mortality (17 studies; RR 0.63, 95% CI 0.43 to 0.92) primarily due to reduced mortality from CMV disease (7 studies; RR 0.26, 95% CI 0.08 to 0.78). Prophylaxis reduced the risk of herpes simplex and herpes zoster disease, bacterial and protozoal infections but not fungal infection, acute rejection or graft loss. Meta-regression showed no significant difference in the relative benefit of treatment (risk of CMV disease or all-cause mortality) by organ transplanted or CMV serostatus; no conclusions were possible for CMV negative recipients of negative organs. In direct comparison studies, ganciclovir was more effective than aciclovir in preventing CMV disease (7 studies; RR 0.37, 95% CI 0.23 to 0.60). Valganciclovir and IV ganciclovir were as effective as oral ganciclovir. AUTHORS' CONCLUSIONS: Prophylaxis with antiviral medications reduces CMV disease and CMV-associated mortality in solid organ transplant recipients. They should be used routinely in CMV positive recipients and in CMV negative recipients of CMV positive organ transplants. PMID- 18425895 TI - Cardioprotective interventions for cancer patients receiving anthracyclines. AB - BACKGROUND: Anthracyclines are among the most effective chemotherapeutic agents in the treatment of numerous malignancies. Unfortunately, their use is limited by a dose-dependent cardiotoxicity. In an effort to prevent this cardiotoxicity, different cardioprotective agents have been studied. OBJECTIVES: The objective of this review was to assess the efficacy of different cardioprotective agents in preventing heart damage in cancer patients treated with anthracyclines. SEARCH STRATEGY: We searched the databases of the Cochrane Central Register of Controlled Trials (CENTRAL, Issue 2, 2007), MEDLINE (1966 to April 2007) and EMBASE (1980 to April 2007). In addition, we handsearched reference lists and conference proceedings of the SIOP and ASCO meetings (1998 to 2006). SELECTION CRITERIA: Randomised controlled trials (RCTs) in which any cardioprotective agent was compared to no additional or placebo therapy in cancer patients (children and adults) receiving anthracyclines. DATA COLLECTION AND ANALYSIS: Two review authors independently performed the study selection, quality assessment and data extraction including adverse effects. MAIN RESULTS: We identified RCTs for seven cardioprotective agents: N-acetylcysteine, phenetylamines, coenzyme Q10, combination of vitamins E and C and N-acetylcysteine, L-carnitine, carvedilol and dexrazoxane (mostly adults with advanced breast cancer). All studies had methodological limitations. For the first six agents, there were too few studies to allow pooling of results. None of the individual studies showed a cardioprotective effect. The nine included studies of dexrazoxane enrolled 1403 patients. The meta-analysis of dexrazoxane showed a statistically significant benefit in favour of dexrazoxane for the occurrence of heart failure (Relative Risk (RR) 0.29, 95% CI 0.20 to 0.41). No evidence was found for a difference in response rate or survival between the dexrazoxane and control group. Only for one adverse effect (abnormal white blood cell count at nadir) a difference in favour of the control group was identified. AUTHORS' CONCLUSIONS: For cardioprotective agents for which pooling was impossible, no definitive conclusions can be made about their efficacy. Dexrazoxane prevents heart damage and no evidence for a difference in response rate or survival between the dexrazoxane and control group was identified. Only for an abnormal white blood cell count at nadir a clearly significant difference in favour of the control group was identified. We conclude that if the risk of cardiac damage is expected to be high, it might be justified to use dexrazoxane in patients with cancer treated with anthracyclines. However, for each individual patient clinicians should weigh the cardioprotective effect of dexrazoxane against the possible risk of adverse effects. PMID- 18425896 TI - Types of urethral catheters for management of short-term voiding problems in hospitalised adults. AB - BACKGROUND: Urinary tract infection (UTI) is the most common hospital acquired infection. The major associated cause is indwelling urinary catheters. Currently there are many types of catheters available. A variety of specialised urethral catheters have been designed to reduce the risk of infection. These include antiseptic impregnated catheters and antibiotic impregnated catheters. Other issues that should be considered when choosing a catheter are ease of use, comfort and cost. OBJECTIVES: The primary objective of this review was to determine the effect of type of indwelling urethral catheter on the risk of urinary tract infection in adults who undergo short-term urinary catheterisation. SEARCH STRATEGY: We searched the Specialised Trials Register of the Cochrane Incontinence Group (searched 11 September 2007). We also examined the bibliographies of relevant articles and contacted catheter manufacturer representatives for trials. SELECTION CRITERIA: All randomised and quasi randomised trials comparing types of indwelling urinary catheters for short-term catheterisation in hospitalised adults. Short-term catheterisation was defined as up to and including fourteen days, or other temporary short-term use as defined by the trialists (for example less than 21 days with data time points at 7 day intervals). DATA COLLECTION AND ANALYSIS: Data were extracted by one reviewer and independently verified by a second reviewer. Disagreements were resolved by discussion. Data were processed as described in the Cochrane Handbook. Where data in trials were not fully reported, clarification was sought directly from the trialists (secondary sources were used to confirm results of one trial). MAIN RESULTS: Twenty three trials met the inclusion criteria involving 5236 hospitalised adults in 22 parallel group trials and 27,878 adults in one large cluster-randomised cross-over trial. The antiseptic catheters were either impregnated with silver oxide or silver alloy. Silver oxide catheters were not associated with a statistically significant reduction in bacteriuria in short term catheterised hospitalised adults but the confidence intervals were wide (RR 0.89, 95% CI 0.68 to 1.15) and these catheters are no longer available. Silver alloy catheters were found to significantly reduce the incidence of asymptomatic bacteriuria (RR 0.54, 95% CI 0.43 to 0.67) in hospitalised adults catheterised for less than one week. At greater than one week of catheterisation the risk of asymptomatic bacteriuria was still reduced with the use of silver alloy catheters (RR 0.64, 95% CI 0.51 to 0.80). The randomised cross-over trial of silver alloy catheters versus standard catheters was excluded from the pooled results because data were not available prior to crossover. The results of this trial indicated benefit from the silver alloy catheters and included an economic analysis that indicated cost savings of between 3.3 per cent and 35.5 per cent. Antibiotic impregnated catheters were compared to standard catheters and found to lower the rate of asymptomatic bacteriuria in the antibiotic group at less than one week of catheterisation for both minocycline and rifampicin (RR 0.36, 95% CI 0.18 to 0.73), and nitrofurazone (RR 0.52, 95% CI 0.34 to 0.78). However, at greater than one week the results were not statistically significant. One of 56 men in the antibiotic impregnated group had a symptomatic UTI compared with 6 of 68 who had standard catheters (RR 0.20, 95% CI 0.03 to 1.63). Three trials compared two different types of standard catheters (defined as catheters that are not impregnated with antiseptics or antibiotics) to investigate infection. Individual trials were too small to show whether or not one type of standard catheter reduced the risk of catheter related urinary tract infection compared to another type of standard catheter. AUTHORS' CONCLUSIONS: The results suggest that the use of silver alloy indwelling catheters for catheterising hospitalised adults short term reduces the risk of catheter acquired urinary tract infection. Further economic evaluation is required to confirm that the reduction of infection compensates for the increased cost of silver alloy catheters.Catheters impregnated with antibiotics are also beneficial in reducing bacteriuria in hospitalised adults catheterised for less than one week but the data were too few to draw conclusions about those catheterised for longer. There was not enough evidence to suggest whether or not any standard catheter was better than another in terms of reducing the risk of urinary tract infection in hospitalised adults catheterised short-term. Siliconised catheters may be less likely to cause urethral side effects in men; however, this result should be interpreted with some caution as the trials were small and the outcome definitions and specific catheters compared varied. PMID- 18425897 TI - Effect of timing of umbilical cord clamping of term infants on maternal and neonatal outcomes. AB - BACKGROUND: Policies for timing of cord clamping vary, with early cord clamping generally carried out in the first 60 seconds after birth, whereas later cord clamping usually involves clamping the umbilical cord greater than one minute after the birth or when cord pulsation has ceased. OBJECTIVES: To determine the effects of different policies of timing of cord clamping at delivery of the placenta on maternal and neonatal outcomes. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group's Trials Register (December 2007). SELECTION CRITERIA: Randomised controlled trials comparing early and late cord clamping. DATA COLLECTION AND ANALYSIS: Two review authors independently assessed trial eligibility and quality and extracted data. MAIN RESULTS: We included 11 trials of 2989 mothers and their babies. No significant differences between early and late cord clamping were seen for postpartum haemorrhage or severe postpartum haemorrhage in any of the five trials (2236 women) which measured this outcome (relative risk (RR) for postpartum haemorrhage 500 mls or more 1.22, 95% confidence interval (CI) 0.96 to 1.55). For neonatal outcomes, our review showed both benefits and harms for late cord clamping. Following birth, there was a significant increase in infants needing phototherapy for jaundice (RR 0.59, 95% CI 0.38 to 0.92; five trials of 1762 infants) in the late compared with early clamping group. This was accompanied by significant increases in newborn haemoglobin levels in the late cord clamping group compared with early cord clamping (weighted mean difference 2.17 g/dL; 95% CI 0.28 to 4.06; three trials of 671 infants), although this effect did not persist past six months. Infant ferritin levels remained higher in the late clamping group than the early clamping group at six months. AUTHORS' CONCLUSIONS: One definition of active management includes directions to administer an uterotonic with birth of the anterior shoulder of the baby and to clamp the umbilical cord within 30-60 seconds of birth of the baby (which is not always feasible in practice). In this review delaying clamping of the cord for at least two to three minutes seems not to increase the risk of postpartum haemorrhage. In addition, late cord clamping can be advantageous for the infant by improving iron status which may be of clinical value particularly in infants where access to good nutrition is poor, although delaying clamping increases the risk of jaundice requiring phototherapy. PMID- 18425898 TI - Substitution treatment of injecting opioid users for prevention of HIV infection. AB - BACKGROUND: Injecting drug users are vulnerable to infection with HIV and other blood borne viruses as a result of collective use of injecting equipment as well as sexual behaviour. OBJECTIVES: To assess the effect of oral substitution treatment for opioid dependent injecting drug users on rates of HIV infections, and high risk behaviours. SEARCH STRATEGY: We searched the Cochrane Central Register of Controlled Trials, MEDLINE, EMBASE and PsycINFO to March 2007. We also searched reference lists of articles, reviews and conference abstracts SELECTION CRITERIA: Studies were required to consider the incidence of risk behaviours, or the incidence of HIV infection related to substitution treatment of opioid dependence. All types of original studies were considered. Two reviewers independently assessed studies for inclusion. DATA COLLECTION AND ANALYSIS: One reviewer extracted data from included studies, assessed quality and confirmed decisions by consulting with all other reviewers. MAIN RESULTS: Thirty three studies, involving 10,400 participants, were included. The majority were not randomised controlled studies and there were problems of confounding and bias. The studies varied in several aspects limiting the extent of quantitative analysis. Studies consistently show that oral substitution treatment for opioid dependent injecting drug users is associated with statistically significant reductions in illicit opioid use, injecting use and sharing of injecting equipment. It is also associated with reductions in the proportion of injecting drug users reporting multiple sex partners or exchanges of sex for drugs or money, but has little effect on condom use. It appears that the reductions in risk behaviours related to drug use do translate into reductions in cases of HIV infection. AUTHORS' CONCLUSIONS: Oral substitution treatment for injecting opioid users reduces drug-related behaviours with a high risk of HIV transmission, but has less effect on sex-related risk behaviours. The lack of data from randomised controlled studies limits the strength of the evidence presented in this review. PMID- 18425899 TI - Clobazam as an add-on in the management of refractory epilepsy. AB - BACKGROUND: Epilepsy effects approximately 1% of the population, with up to 30% of patients continuing to have seizures despite antiepileptic drug treatment. OBJECTIVES: To assess the efficacy and tolerability of clobazam when used as an add-on therapy for patients with refractory partial onset or generalised onset seizures. SEARCH STRATEGY: We searched the following on 22nd March 2007: (a) The Cochrane Epilepsy Group Specialised Register; (b) The Cochrane Central Register of Controlled Trials (CENTRAL); (c) MEDLINE; (d) EMBASE; (e) Database of Abstracts of Reviews of Effectiveness (DARE); (f) American College of Physicians Journals; (g) BIOSIS. SELECTION CRITERIA: Randomised trials of add-on clobazam, with adequate methods of allocation concealment, recruiting patients with drug refractory partial or generalised onset seizures, with a minimum treatment period of eight weeks. DATA COLLECTION AND ANALYSIS: Two review authors independently selected trials for inclusion and extracted relevant data. The following outcomes were assessed: 50% or greater reduction in seizures; seizure freedom; treatment withdrawal and adverse effects. MAIN RESULTS: Four cross-over studies, representing 196 participants, were included. However, due to significant methodological heterogeneity and differences in outcome measures it was not possible to summarise data in a meta-analysis. Only two of the studies reported a 50% or greater seizure reduction compared to placebo; 57.7% and 52.4%. Side effects were only described in two of the studies, reportedly present in 36% and 85% of patients. AUTHORS' CONCLUSIONS: Clobazam as an add-on treatment may reduce seizure frequency and may be most effective in partial onset seizures. However, it is not clear who will best benefit and over what time-frame. A large scale, randomised controlled trial conducted over a greater period of time, incorporating subgroups with differing seizure types, is required to inform clinical practice. PMID- 18425900 TI - Palliative endobronchial brachytherapy for non-small cell lung cancer. AB - BACKGROUND: Non-small cell lung cancers (NSCLC) constitutes about 80% of all lung cancer cases. Although surgery is the only curative treatment of NSCLC, fewer than 20% of tumors can be radically resected. Radiotherapy is one of the main treatment modalities in lung cancer, contributing to both its cure and palliation. Endobronchial brachytherapy (EBB) has been used as one approach to improve local control either alone or in combination with other treatments. OBJECTIVES: To assess the effectiveness of palliative EBB in increasing survival and to control thoracic symptoms in patients with advanced NSCLC compared with external beam radiation therapy (EBRT) or other alternative endoluminal treatments. SEARCH STRATEGY: The Cochrane Central Register of Controlled Trials (CENTRAL), MEDLINE, EMBASE and other databases were searched, as were reference lists and handsearching of selected journals and conference proceedings. SELECTION CRITERIA: Randomized controlled trials (RCTs) comparing different regimens of palliative EBB with EBRT or other endobronchial interventions in patients with advanced NSCLC. DATA COLLECTION AND ANALYSIS: Thirteen RCTs were included. There were important differences in the doses of radiotherapy investigated, patient characteristics and the outcomes measured. Because of this heterogeneity no meta-analysis was attempted. MAIN RESULTS: We found trials comparing EBB to EBRT alone, EBB plus EBRT to EBRT alone, EBB plus chemotherapy to EBB alone, EBB to Nd-YAG laser and comparisons between diverse fractionation schedules of high dose rate EBB. From the heterogeneous information obtained from several small RCTs, we concluded that EBRT alone is more effective for palliation of NSCLC symptoms than EBB alone. Our findings did not provide conclusive evidence to recommend EBB plus EBRT to relieve symptoms compared to EBRT alone. Overall, for the primary endpoint of survival there was no evidence of benefit for EBB compared to EBRT and Nd-YAG laser or for the combination of EBB with chemotherapy. Additionally, findings from one trial suggested that twice 7.4 Gy was superior to the four times per week 3.8 Gy schedule for mean time of local control and fatal haemoptysis. No significant differences were found for fatal haemoptysis as an adverse event of EBB. AUTHORS' CONCLUSIONS: The evidence did not provide conclusive results that EBB plus EBRT improved symptom relief over EBRT alone. We were not able to provide conclusive evidence to recommend EBB with EBRT, chemotherapy or Nd-YAG laser. For patients previously treated by EBRT who are symptomatic from recurrent endobronchial central obstruction, EBB may be considered in selected cases. PMID- 18425901 TI - Interventions for alopecia areata. AB - BACKGROUND: Alopecia areata is a disorder in which there is loss of hair causing patches of baldness but with no scarring of the affected area. It can affect the entire scalp (alopecia totalis) or cause loss of all body hair (alopecia universalis). It is a relatively common condition affecting 0.15% of the population. Although in many cases it can be a self-limiting condition, nevertheless hair loss can often have a severe social and emotional impact. OBJECTIVES: To assess the effects of interventions used in the management of alopecia areata, alopecia totalis and alopecia universalis. SEARCH STRATEGY: We searched the Cochrane Skin Group Specialised Register in February 2006, the Cochrane Central Register of Controlled Clinical Trials (The Cochrane Library Issue 1, 2006), MEDLINE (from 2003 to February 2006), EMBASE (from 2005 to February 2006), PsycINFO (from 1806 to February 2006), AMED (Allied and Complementary Medicine, from 1985 to February 2006), LILACS (Latin American and Caribbean Health Science Information database, from 1982 to February 2006), and reference lists of articles. We also searched online trials registries for ongoing trials. SELECTION CRITERIA: Randomised controlled trials that evaluated the effectiveness of both topical and systemic interventions for alopecia areata, alopecia totalis, and alopecia universalis. DATA COLLECTION AND ANALYSIS: Two authors assessed trial quality and extracted the data. We contacted trial authors for more information. We collected adverse effects information from the included trials. MAIN RESULTS: Seventeen trials were included with a total of 540 participants. Each trial included from 6 to 85 participants and they assessed a range of interventions that included topical and oral corticosteroids, topical ciclosporin, photodynamic therapy and topical minoxidil. Overall, none of the interventions showed significant treatment benefit in terms of hair growth when compared with placebo. We did not find any studies where the participants self assessed their hair growth or quality of life. AUTHORS' CONCLUSIONS: Few treatments for alopecia areata have been well evaluated in randomised trials. We found no RCTs on the use of diphencyprone, dinitrochlorobenzene, intralesional corticosteroids or dithranol although they are commonly used for the treatment of alopecia areata. Similarly although topical steroids and minoxidil are widely prescribed and appear to be safe, there is no convincing evidence that they are beneficial in the long-term. Most trials have been reported poorly and are so small that any important clinical benefits are inconclusive. There is a desperate need for large well conducted studies that evaluate long-term effects of therapies on quality of life. Considering the possibility of spontaneous remission especially for those in the early stages of the disease, the options of not being treated therapeutically or, depending on individual preference wearing a wig may be alternative ways of dealing with this condition. PMID- 18425902 TI - Antiepileptic drugs for preventing seizures in people with brain tumors. AB - BACKGROUND: Seizures can present at any time before or after diagnosis of a brain tumor. The risk of seizures varies by tumor type and its location in the brain. For a long time we believed that preventing seizures with antiepileptic drugs (seizure prophylaxis) was effective and necessary, but the supporting evidence was little and mixed. Such evidence was the basis for previous reviews to conclude that seizure prophylaxis was ineffective in people with brain tumors. OBJECTIVES: To estimate the effectiveness of seizure prophylaxis in people with brain tumors, and to estimate the adverse event rates in the identified clinical trials. SEARCH STRATEGY: A search strategy that included free-text and MeSH terms in LILACS, EMBASE, PubMed, CENTRAL, and The Cochrane Library (1966 to 2007). SELECTION CRITERIA: Controlled clinical trials with random allocation, blinded or unblinded, and placebo or observation in the control groups. DATA COLLECTION AND ANALYSIS: We screened the articles, extracted the data, and rated the validity of each trial to assess the risk of bias. Our primary outcome was the occurrence of a first seizure. The secondary outcome was adverse events. We pooled the aggregate data for each outcome into a random-effects model meta-analysis using the relative risk (RR). For adverse events, we also included the number needed to harm (NNH) using the absolute risk increase to compute the NNH. MAIN RESULTS: There was no difference between the treatment interventions and the control groups in preventing a first seizure in participants with brain tumors. The risk of an adverse event was higher for those on antiepileptic drugs than for participants not on antiepileptic drugs (NNH 3; RR 6.10, 95% CI 1.10 to 34.63; P = 0.046). AUTHORS' CONCLUSIONS: The evidence is neutral, neither for nor against seizure prophylaxis, in people with brain tumors. These conclusions apply only for the antiepileptic drugs phenytoin, phenobarbital, and divalproex sodium. The decision to start an antiepileptic drug for seizure prophylaxis is ultimately guided by assessment of individual risk factors and careful discussion with patients. PMID- 18425903 TI - Danshen (Chinese medicinal herb) preparations for acute myocardial infarction. AB - BACKGROUND: Acute myocardial infarction (AMI) is the most important cause of morbidity from ischaemic heart disease, and is among the leading causes of death in the western world. Danshen, a Chinese herbal medicine, is widely used in China for treatment of several diseases, including AMI. OBJECTIVES: To assess the effects (both benefits and harms) of danshen preparations for AMI. SEARCH STRATEGY: We searched the Cochrane Central Register of Controlled Trials (CENTRAL) on The Cochrane Library (issue 4, 2006), MEDLINE (1966-2006), EMBASE (1980-2006), and the Chinese Biomedical Database (CBM) (1982-2006). We also handsearched 75 Chinese medical journals. SELECTION CRITERIA: Randomised controlled trials (RCTs) lasting at least 7 days were sought. Since it seemed evident that few RCTs were available, we also considered other controlled studies. DATA COLLECTION AND ANALYSIS: Eligibility and trial quality were assessed by three reviewers. MAIN RESULTS: Six studies comprised of 2368 participants were included. Only one trial was judged to be a genuine RCT and showed no statistically significant difference in reduction of total mortality (Peto OR 0.55, 95% CI 0.23 to 1.32), but a quasi-RCT reported a reduced total mortality (Peto OR 0.42, 95% CI 0.23 to 0.77). Pooling these trials yielded an approximate halving of mortality in those patients treated with danshen preparations plus usual care compared with usual care alone (Peto OR 0.46, 95% CI 0.28 to 0.75). AUTHORS' CONCLUSIONS: The evidence to support use of danshen preparations is too weak to make any judgement about its effects. Evidence from RCTs is insufficient and of low quality. The safety of danshen preparations is unproven, although some adverse events have been reported. More evidence from high quality trials is needed to support the clinical use of danshen preparations. PMID- 18425904 TI - Interventions for treating wrist fractures in children. AB - BACKGROUND: Approximately a third of all fractures in children occur at the wrist, usually from falling onto an outstretched hand. OBJECTIVES: We aimed to evaluate removable splintage versus plaster casts (requiring removal by a specialist) for undisplaced compression (buckle) fractures; cast length and position; and the role of surgical fixation for displaced wrist fractures in children. SEARCH STRATEGY: We searched the Cochrane Bone, Joint and Muscle Trauma Group Specialised Register (October 2007), the Cochrane Central Register of Controlled Trials (The Cochrane Library 2007, Issue 4), MEDLINE (from 1966), EMBASE (from 1988), CINAHL (from 1982) and reference lists of articles. Date of last search October 2007. SELECTION CRITERIA: Any randomised or quasi-randomised controlled trials comparing types and position of casts and the use of surgical fixation for distal radius fractures in children. DATA COLLECTION AND ANALYSIS: Two authors performed trial selection. All three authors independently assessed methodological quality and extracted data. MAIN RESULTS: The 10 included trials, involving 827 children, were of variable quality.Four trials compared removable splintage versus the traditional below-elbow cast in children with buckle fractures. There was no short-term deformity recorded in all four trials and, in one trial, no refracture at six months. The Futura splint was cheaper to use; a removable plaster splint was less restrictive to wear enabling more children to bathe and participate in other activities, and the option preferred by children and parents; the soft bandage was more comfortable, convenient and less painful to wear; home-removable plaster casts removed by parents did not result in significant differences in outcome but were strongly favoured by parents. Two trials found below-elbow versus above-elbow casts did not increase redisplacement of reduced fractures or cast-related complications, were less restrictive during use and avoided elbow stiffness. One trial evaluating the effect of arm position in above-elbow casts found no effect on deformity. Three trials found that percutaneous wiring significantly reduced redisplacement and remanipulation but one of these found no advantage in function at three months. AUTHORS' CONCLUSIONS: Limited evidence supports the use of removable splintage for buckle fractures and challenges the traditional use of above-elbow casts after reduction of displaced fractures. Although percutaneous wire fixation prevents redisplacement, the effects on longer term outcomes including function are not established. Further research is warranted on the optimum approach, including splintage, for buckle fractures; and on the use of below-elbow casts and indications for surgery for displaced wrist fractures in children. PMID- 18425905 TI - Vaccines for preventing influenza in healthy children. AB - BACKGROUND: The consequences of influenza in children and adults are mainly absenteeism from school and work. However, the risk of complications is greatest in children and people over 65 years old. OBJECTIVES: To appraise all comparative studies evaluating the effects of influenza vaccines in healthy children; assess vaccine efficacy (prevention of confirmed influenza) and effectiveness (prevention of influenza-like illness) and document adverse events associated with influenza vaccines. SEARCH STRATEGY: We searched the Cochrane Central Register of Controlled Trials (CENTRAL) (The Cochrane Library 2007, issue 3); OLD MEDLINE (1950 to 1965); MEDLINE (1966 to September 2007); EMBASE (1974 to September 2007); Biological Abstracts (1969 to September 2007); and Science Citation Index (1974 to September 2007). SELECTION CRITERIA: Randomised controlled trials (RCTs), cohort and case-control studies of any influenza vaccine in healthy children under 16 years of age. DATA COLLECTION AND ANALYSIS: Two review authors independently assessed trial quality and extracted data. MAIN RESULTS: Fifty-one studies with 294,159 observations were included. Sixteen RCTs and 18 cohort studies were included in the analysis of vaccine efficacy and effectiveness. From RCTs, live vaccines showed an efficacy of 82% (95% confidence interval (CI) 71% to 89%) and an effectiveness of 33% (95% CI 28% to 38%) in children older than two compared with placebo or no intervention. Inactivated vaccines had a lower efficacy of 59% (95% CI 41% to 71%) than live vaccines but similar effectiveness: 36% (95% CI 24% to 46%). In children under two, the efficacy of inactivated vaccine was similar to placebo. Variability in study design and presentation of data was such that a meta-analysis of safety outcome data was not feasible. Extensive evidence of reporting bias of safety outcomes from trials of live attenuated vaccines impeded meaningful analysis. AUTHORS' CONCLUSIONS: Influenza vaccines are efficacious in children older than two but little evidence is available for children under two. There was a marked difference between vaccine efficacy and effectiveness. No safety comparisons could be carried out, emphasizing the need for standardisation of methods and presentation of vaccine safety data in future studies. It was surprising to find only one study of inactivated vaccine in children under two years, given current recommendations to vaccinate healthy children from six months old in the USA and Canada. If immunisation in children is to be recommended as a public health policy, large-scale studies assessing important outcomes and directly comparing vaccine types are urgently required. PMID- 18425906 TI - Multidisciplinary rehabilitation programmes following joint replacement at the hip and knee in chronic arthropathy. AB - BACKGROUND: Joint replacements are common procedures and treatment of choice for those with intractable joint pain and disability arising from arthropathy of the hip or knee. Multidisciplinary rehabilitation is considered integral to the outcome of joint replacement. OBJECTIVES: To assess the evidence for effectiveness of multidisciplinary rehabilitation on activity and participation in adults following hip or knee joint replacement for chronic arthropathy. SEARCH STRATEGY: We searched the Cochrane Musculoskeletal Group Trials Register, the Cochrane Central Register of Controlled Trials, MEDLINE, EMBASE and CINAHL up to September 2006. SELECTION CRITERIA: Randomised controlled trials (RCTs) that compared organised multidisciplinary rehabilitation with routine services following hip or knee replacement, and included outcome measures of activity and participation in accordance with the International Classification of Functioning, Health and Disability (ICF). DATA COLLECTION AND ANALYSIS: Four authors independently extracted data and assessed methodological quality of included trials. MAIN RESULTS: Five trials (619 participants) met the inclusion criteria; two addressed inpatient rehabilitation (261 participants) and three (358 participants) home-based settings. There were no trials addressing outpatient centre-based programmes. Pooling of data was not possible due to differences in study design and outcomes used. Methodological assessment showed all trials were of low quality. For inpatient settings early commencement of rehabilitation and clinical pathways led to more rapid attainment of functional milestones (disability) (Functional Independence Measure (FIM) transfer WMD 0.5, 95% CI 0.15, 0.85, number needed to treat to benefit (NNTB) = 6, FIM ambulation WMD 1.55 (95%CI 0.96, 2.14), NNTB = 3), shorter hospital stay, fewer post-operative complications and reduced costs in the first three to four months. Home-based multidisciplinary care improved functional gain (Oxford Hip Score (OHS) WMD at 6 months -7.00 (95%CI -10.36, -3.64), NNT = 2 and quality of life (QoL) and reduced hospital stay in the medium term (six months). No trials addressed longer-term outcomes following hip replacement only. AUTHORS' CONCLUSIONS: Based on the heterogeneity and the low quality of the included trials that precluded pooled meta-analysis, there is silver level evidence that following hip or knee joint replacement, early multidisciplinary rehabilitation can improve outcomes at the level of activity and participation. The optimal intensity, frequency and effects of rehabilitation over a longer period and associated social costs need further study. Future research should focus on improving methodological and scientific rigour of clinical trials, and use of standardised outcome measures, so that results can be pooled for statistical analysis. PMID- 18425907 TI - Interventions for replacing missing teeth: treatment of perimplantitis. AB - BACKGROUND: One of the key factors for the long-term success of oral implants is the maintenance of healthy tissues around them. Bacterial plaque accumulation induces inflammatory changes in the soft tissues surrounding oral implants and it may lead to their progressive destruction (perimplantitis) and ultimately to implant failure. Different treatment strategies for perimplantitis have been suggested, however it is unclear which are the most effective. OBJECTIVES: To identify the most effective interventions for treating perimplantitis around osseointegrated dental implants. SEARCH STRATEGY: We searched the Cochrane Oral Health Group's Trials Register, the Cochrane Central Register of Controlled Trials (CENTRAL), MEDLINE and EMBASE. Handsearching included several dental journals. We checked the bibliographies of the identified randomised controlled trials (RCTs) and relevant review articles for studies outside the handsearched journals. We wrote to authors of all identified RCTs, to more than 55 dental implant manufacturers and an Internet discussion group to find unpublished or ongoing RCTs. No language restrictions were applied. The last electronic search was conducted on 9 January 2008. SELECTION CRITERIA: All RCTs comparing agents or interventions for treating perimplantitis around dental implants. DATA COLLECTION AND ANALYSIS: Screening of eligible studies, assessment of the methodological quality of the trials and data extraction were conducted in duplicate and independently by two review authors. We contacted the authors for missing information. Results were expressed as random-effects models using weighted mean differences for continuous outcomes and risk ratios for dichotomous outcomes with 95% confidence intervals (CI). Heterogeneity was to be investigated including both clinical and methodological factors. MAIN RESULTS: Ten eligible trials were identified, but three were excluded. The following procedures were tested: (1) use of local antibiotics versus ultrasonic debridement; (2) benefits of adjunctive local antibiotics to debridement; (3) different techniques of subgingival debridement; (4) laser versus manual debridement and chlorhexidine irrigation/gel; (5) systemic antibiotics plus resective surgery plus two different local antibiotics with and without implant surface smoothening; and (6) nanocrystalline hydroxyapatite versus Bio-Oss and resorbable barriers. Follow up ranged from 3 months to 2 years. The only statistically significant differences were observed in two trials judged to be at high risk of bias. After 4 months, adjunctive local antibiotics to manual debridement in patients who lost at least 50% of the bone around implants showed improved mean probing attachment levels (PAL) of 0.61 mm and reduced probing pockets depths (PPD) of 0.59 mm. After 6 months, patients with perimplant infrabony defects > 3 mm treated with Bio-Oss and resorbable barriers gained 0.5 mm more PAL (borderline difference) and PPD than patients treated with a nanocrystalline hydroxyapatite. AUTHORS' CONCLUSIONS: There is very little reliable evidence suggesting which could be the most effective interventions for treating perimplantitis. This is not to say that currently used interventions are not effective. The use of local antibiotics in addition to manual subgingival debridement was associated with a 0.6 mm additional improvement for PAL and PPD over a 4-month period in patients affected by severe forms of perimplantitis. After 6 months both augmentation therapies appeared to be successful but improved PAL and PPD (0.5 mm) were obtained when using Bio-Oss with resorbable barriers. In four trials, the control therapy which basically consisted of a simple subgingival mechanical debridement seemed to be sufficient to achieve results similar to the more complex and expensive therapies. Sample sizes were very small and follow up too short, therefore these conclusions have to be considered with great caution. Larger well-designed RCTs are needed. PMID- 18425908 TI - Excisional surgery versus ablative surgery for ovarian endometriomata. AB - BACKGROUND: Endometriomata are endometriotic deposits within the ovary. The surgical management of these blood filled cysts is controversial. The laparoscopic approach to the management of endometriomata is favoured over a laparotomy approach as it offers the advantage of a shorter hospital stay, faster patient recovery and decreased hospital costs. Currently the commonest procedures for the treatment of ovarian endometriomata are either excision of the cyst capsule or drainage and electrocoagulation of the cyst wall. OBJECTIVES: The objective of this review was to determine the most effective technique of treating an ovarian endometrioma; either excision of the cyst capsule or drainage and electrocoagulation of the cyst wall. The end-points assessed were the relief of pain, recurrence of the endometrioma, recurrence of symptoms and in women desiring to conceive the subsequent pregnancy rate, either spontaneous or as part of fertility treatment. SEARCH STRATEGY: The reviewers searched the Cochrane Menstrual Disorders and Subfertility Group specialised register of trials (searched 3rd March 2007), the Cochrane Register of Controlled Trials (The Cochrane Library, Issue 3, 2007), MEDLINE (1966-August 2007), EMBASE (1980- March 2007) and reference lists of articles, the handsearching of relevant journals and conference proceedings and by contacting leaders in the field of endoscopic surgery throughout the world. The Cochrane Menstrual Disorders and Subfertility Group Trials Register is based on regular searches of MEDLINE, EMBASE, CINHAL and CENTRAL. SELECTION CRITERIA: Randomised controlled trials of excision of the cyst capsule versus drainage and electrocoagulation of the cyst in the management of ovarian endometriomata. DATA COLLECTION AND ANALYSIS: Reviewers assessed eligibility and trial quality. MAIN RESULTS: No randomised studies of the management of endometriomata by laparotomy were found. Two randomised studies of the laparoscopic management of ovarian endometriomata of greater than 3cm in size, for the primary symptom of pain were included. Laparoscopic excision of the cyst wall of the endometrioma was associated with a reduced recurrence rate of the symptoms of dysmenorrhea (OR 0.15 CI 0.06-0.38), dyspareunia (OR 0.08 CI 0.01 0.51) and non-menstrual pelvic pain (OR 0.10 CI 0.02-0.56), a reduced rate of recurrence of the endometrioma (OR 0.41 CI 0.18-0.93) and with a reduced requirement for further surgery (OR 0.21 CI 0.05-0.79) than surgery to ablate the endometrioma. For those women subsequently attempting to conceive it was also associated with a subsequent increased spontaneous pregnancy rate in women who had documented prior sub-fertility (OR 5.21 CI 2.04-13.29). A further randomised study was identified that demonstrated an increased ovarian follicular response to gonadotrophin stimulation for women who had undergone excsional surgery when compared to ablative surgery (WMD 0.6 CI 0.04-1.16). There is insufficient evidence to favour excisional surgery over ablative surgery with respect to the chance of pregnancy after controlled ovarian stimulation and intra-uterine insemination (OR 1.40 CI 0.47-4.15) . AUTHORS' CONCLUSIONS: There is good evidence that excisional surgery for endometriomata provides for a more favourable outcome than drainage and ablation with regard to the recurrence of the endometrioma, recurrence of pain symptoms, and in women who were previously subfertile, subsequent spontaneous pregnancy . Consequently this approach should be the favoured surgical approach. However in women who may subsequently may undergo fertility treatment insufficient evidence exists to determine the favoured surgical approach. PMID- 18425909 TI - Low level laser therapy for nonspecific low-back pain. AB - BACKGROUND: Low-back pain (LBP) is a major health problem and a major cause of medical expenses and disablement. Low level laser therapy (LLLT) can be used to treat musculoskeletal disorders such as back pain. OBJECTIVES: To assess the effects of LLLT in patients with non-specific LBP. SEARCH STRATEGY: We searched CENTRAL (The Cochrane Library 2005, Issue 2), MEDLINE, CINAHL, EMBASE, AMED and PEDro from their start to November 2007 with no language restrictions. We screened references in the included studies and in reviews and conducted citation tracking of identified RCTs and reviews using Science Citation Index. We also contacted content experts. SELECTION CRITERIA: Randomised controlled clinical trials (RCTs) investigating LLLT to treat non-specific low-back pain were included. DATA COLLECTION AND ANALYSIS: Two authors independently assessed methodological quality using the criteria recommended by the Cochrane Back Review Group and extracted data. Studies were qualitatively and quantitatively analysed according to Cochrane Back Review Group guideline. MAIN RESULTS: Seven heterogeneous English language RCTs with reasonable quality were included. Three small studies (168 people) separately showed statistically significant but clinically unimportant pain relief for LLLT versus sham therapy for sub-acute and chronic low-back pain at short-term and intermediate-term follow-up (up to six months). One study (56 people) showed that LLLT was more effective than sham at reducing disability in the short term. Three studies (102 people) reported that LLLT plus exercise were not better than exercise, with or without sham in the short-term in reducing pain or disability. Two studies (90 people) reported that LLLT was not more effective than exercise, with or without sham in reducing pain or disability in the short term. Two small trials (151 people) independently found that the relapse rate in the LLLT group was significantly lower than in the control group at the six-month follow-up. No side effects were reported. AUTHORS' CONCLUSIONS: Based on the heterogeneity of the populations, interventions and comparison groups, we conclude that there are insufficient data to draw firm conclusions on the clinical effect of LLLT for low-back pain. There is a need for further methodologically rigorous RCTs to evaluate the effects of LLLT compared to other treatments, different lengths of treatment, wavelengths and dosages. PMID- 18425910 TI - Oestrogens for preventing recurrent urinary tract infection in postmenopausal women. AB - BACKGROUND: Recurrent urinary tract infection (RUTI) is defined as three episodes of urinary tract infection (UTI) in the previous 12 months or two episodes in the last six months. The main factors associated with RUTI in postmenopausal women are vesical prolapse, cystocoele, post-voidal residue and urinary incontinence, all associated with a decrease in oestrogen. The use of oestrogens to prevent RUTI has been proposed. OBJECTIVES: To estimate the efficacy and safety of oral or vaginal oestrogens for preventing RUTI in postmenopausal women. SEARCH STRATEGY: We searched the Cochrane Renal Group's specialised register, the Cochrane Central Register of Controlled Trials (CENTRAL), MEDLINE (from 1950), EMBASE (from 1980), reference lists of articles without language restriction. Date of last search: February 2007. SELECTION CRITERIA: Randomised controlled trials (RCTs) in which postmenopausal women (more than 12 months since last menstrual period) diagnosed with RUTI received any type of oestrogen (oral , vaginal) versus placebo or any other intervention were included. DATA COLLECTION AND ANALYSIS: Authors extracted data and assessed quality. Statistical analyses were performed using the random effects model and the results expressed as relative risk (RR) for dichotomous outcomes or mean difference (WMD) for continuous data with 95% confidence intervals (CI). MAIN RESULTS: Nine studies (3345 women) were included. Oral oestrogens did not reduce UTI compared to placebo (4 studies, 2798 women: RR 1.08, 95% CI 0.88 to 1.33). Vaginal oestrogens versus placebo reduced the number of women with UTIs in two small studies using different application methods. The RR for one was 0.25 (95% CI 0.13 to 0.50) and 0.64 (95% CI 0.47 to 0.86) in the second. Two studies compared oral antibiotics versus vaginal oestrogens (cream (1), pessaries (1)). There was very significant heterogeneity and the results could not be pooled. Vaginal cream reduced the proportion of UTIs compared to antibiotics in one study and in the second study antibiotics were superior to vaginal pessaries. Adverse events for vaginal oestrogens were breast tenderness, vaginal bleeding or spotting, nonphysiologic discharge, vaginal irritation, burning and itching. AUTHORS' CONCLUSIONS: Based on only two studies comparing vaginal oestrogens to placebo, vaginal oestrogens reduced the number of UTIs in postmenopausal women with RUTI, however this varied according to the type of oestrogen used and the treatment duration. PMID- 18425912 TI - Omega-3 fatty acids for bipolar disorder. AB - BACKGROUND: Bipolar disorder is a complex psychiatric disorder and is amongst the top thirty causes of worldwide disability. Mood stabilisers are the primary pharmacological intervention, both in the treatment of acute episodes and in prophylaxis. There is, however, mounting evidence that dietary supplementation with omega-3 fatty acids may be beneficial in psychiatric conditions, particularly those involving disturbances of mood. OBJECTIVES: To review the efficacy of omega-3 fatty acids as either a monotherapy or an adjunctive treatment for bipolar disorder. SEARCH STRATEGY: Electronic searches of the following databases were performed: CCDANCTR-Studies and CCDANCTR-References were searched on 12/2/2008, Supplementary searches were carried out on Biological Abstracts, CINAHL, The Cochrane Library, CCDAN Register, EMBASE, MEDLINE, and PsycINFO. The search strategy also included cited reference searching, personal contact with all authors of studies initially included and contact with the omega 3 producing pharmaceutical companies. SELECTION CRITERIA: All relevant randomised controlled trials were included in the review. Studies involving males and females of all ages with a diagnosis of bipolar disorder qualified for inclusion. Studies using any type or dose of omega-3 fatty acid treatment as monotherapy or in addition to standard pharmacotherapy were eligible. The primary outcome was symptom severity; and secondary outcomes were adverse effects, dropout and satisfaction with treatment. DATA COLLECTION AND ANALYSIS: Two review authors independently inspected the citations identified from the search. Potentially relevant abstracts were identified and full papers ordered and reassessed for inclusion and methodological quality. All relevant data were extracted. The weighted mean difference (WMD) was used for continuous outcome data, with 95% confidence intervals (CI). MAIN RESULTS: Five studies met inclusion criteria for the review, however, methodological quality was highly variable. Only one study, involving 75 participants, provided data for analysis, and showed a benefit of active treatment over control for depression symptom levels (WMD -3.93, 95% CI 7.00 to -0.86)and Clinical Global Impression scores (WMD -0.75, 95% CI -1.33 to 0.17) but not for mania (WMD -2.81, 95% CI -7.68 to 1.90). No serious adverse effects were reported in the five studies. The pattern of dropout was highly variable between studies. AUTHORS' CONCLUSIONS: Results from one study showed positive effects of omega-3 as an adjunctive treatment for depressive but not manic symptoms in bipolar disorder. These findings must be regarded with caution owing to the limited data available. There is an acute need for well-designed and executed randomised controlled trials in this field. PMID- 18425913 TI - Therapeutic exercise for people with amyotrophic lateral sclerosis or motor neuron disease. AB - BACKGROUND: Despite the high incidence of muscle weakness in individuals with amyotrophic lateral sclerosis (ALS) or motor neuron disease (MND), the effects of exercise in this population are not well understood. OBJECTIVES: The objective was to systematically review randomised and quasi-randomised studies of exercise for people with ALS or MND. SEARCH STRATEGY: We searched the Cochrane Neuromuscular Disease Group Trials Register, EMBASE (January 1980 to August 2007), LILACS (January 1982 to August 2007), MEDLINE (January 1966 to August 2007), Cochrane Central Register of Controlled Trials (CENTRAL), PEDro (January 1980 to August 2007), AMED (January 1985 to August 2007), HealthSTAR (January 1975 to August 2007), CINAHL (January 1982 to August 2007). We also searched Dissertation Abstracts, inspected the reference lists of all papers selected for review and contacted the authors with expertise in the field. SELECTION CRITERIA: We included randomised or quasi-randomised controlled trials of people with a diagnosis of definite, probable, probable with laboratory support, or possible amyotrophic lateral sclerosis, as defined by the El Escorial criteria. We included progressive resistance or strengthening exercise and endurance or aerobic exercise. The control condition was no exercise or standard rehabilitation management. Our primary outcome measure was improvement in functional ability, decrease in disability or reduction in rate of decline as measured by a validated outcome tool at three months. Our secondary outcome measures were improvement in psychological status or quality of life, decrease in fatigue, increase in, or reduction in rate of decline of muscle strength (strengthening or resistance studies), increase in, or reduction in rate of decline of aerobic endurance (aerobic or endurance studies) at three months and frequency of adverse effects. DATA COLLECTION AND ANALYSIS: Two review authors independently assessed trial quality and extracted the data. The authors of the papers were contacted to obtain information not available in the published articles. MAIN RESULTS: We identified two randomised controlled trials that met our inclusion criteria. The first examined the effects of a twice-daily exercise program of moderate load, endurance exercise versus "usual activities" in 25 people with ALS. The second examined the effects of thrice weekly moderate load and moderate intensity resistance exercises compared to usual care (stretching exercises) in 27 people with ALS. After three months, when the results of the two trials were combined, there was a significant weighted mean improvement in the Amyotrophic Lateral Sclerosis Functional Rating Scale (ALSFRS) measure of function in the exercise compared with the control groups (3.21, 95% confidence interval 0.46 to 5.96) in favour of the exercise group. No statistically significant differences in quality of life, fatigue or muscle strength were found. AUTHORS' CONCLUSIONS: The only studies detected were too small to determine to what extent strengthening exercises for people with ALS are beneficial, or whether exercise is harmful. There is a complete lack of randomised or quasi-randomised clinical trials examining aerobic exercise in this population. More research is needed. PMID- 18425911 TI - Antiangiogenic therapy with anti-vascular endothelial growth factor modalities for neovascular age-related macular degeneration. AB - BACKGROUND: Age-related macular degeneration (AMD) is a common cause of severe vision loss in people 55 years and older. OBJECTIVES: The objective of this review was to investigate the effects of anti-VEGF (vascular endothelial growth factor) modalities for treating neovascular AMD. SEARCH STRATEGY: We searched CENTRAL, MEDLINE, EMBASE and LILACS. We handsearched ARVO abstracts for 2006, 2007 for ongoing trials. SELECTION CRITERIA: We included randomized controlled trials (RCTs). DATA COLLECTION AND ANALYSIS: Two review authors independently extracted data. We contacted trial authors for additional data. We summarized outcomes as relative risks (RR), number needed to treat (NNT) and weighted mean differences. MAIN RESULTS: We included five RCTs of good methodological quality. All five trials were conducted by pharmaceutical companies. An intention-to-treat analysis using the last observation carried forward method was done in most trials. Two trials compared pegaptanib versus sham. One trial compared ranibizumab versus sham, another compared ranibizumab/sham verteporfin PDT versus verteporfin PDT/sham ranibizumab, and the final trial compared ranibizumab plus verteporfin PDT versus verteporfin PDT alone. Fewer patients treated with pegaptanib lost 15 or more letters of visual acuity at one year follow-up compared to sham (pooled relative risk (RR) 0.71; 95% confidence interval (CI) 0.61 to 0.84). The NNT was 6.67 (95% CI 4.35 to 14.28) for 0.3 mg pegaptanib, 6.25 (95% CI 4.17 to 12.5) for 1 mg pegaptanib and 14.28 (95% CI 6.67 to 100) for 3 mg pegaptanib. In a trial of ranibizumab versus sham, RR for loss of 15 or more letters visual acuity at one year was 0.14 (95% CI 0.1 to 0.22) in favour of ranibizumab. The NNT was 3.13 (95% CI 2.56 to 3.84) for 0.3 mg ranibizumab and 3.13 (95% CI 2.56 to 3.84) for 0.5 mg ranibizumab. In a trial of ranibizumab versus verteporfin PDT, RR for loss of 15 or more letters at one year was 0.13 (95% CI 0.07 to 0.23) favouring ranibizumab. The NNT was 3.33 (95% CI 2.56 to 4.76) for 0.3 mg ranibizumab and 3.12 (95% CI 2.43 to 4.17) for 0.5 mg ranibizumab. In another trial of combined ranibizumab plus verteporfin PDT versus verteporfin PDT, RR for loss of 15 or more letters at one year favoured combined therapy (RR 0.3 (95% CI 0.15 to 0.60). The NNT was 4.35 (95% CI 2.78 to 11.11). Pooled RR for gain of 15 or more letters visual acuity at one year was 5.81 (95% CI 3.29 to 10.26) for ranibizumab versus sham, 6.79 (95% CI 3.41 to 13.54) for ranibizumab/sham verteporfin PDT versus verteporfin PDT/sham ranibizumab, and 4.44 (95% CI 1.40 to 14.08) for ranibizumab plus verteporfin PDT versus verteporfin PDT. Frequency of endophthalmitis in included studies was between 0.7% to 4.7% with ranibizumab and 1.3% with pegaptanib. Improvement in vision specific quality of life was reported for both treatments. AUTHORS' CONCLUSIONS: Pegaptanib and ranibizumab reduce the risk of visual acuity loss in patients with neovascular AMD. Ranibizumab causes gains in visual acuity in many eyes. Quality of life and cost will be important for treatment decisions. Other agents blocking VEGF are being tested in ongoing trials. PMID- 18425914 TI - Surfactant for pulmonary hemorrhage in neonates. AB - BACKGROUND: In the late 1960's and 1970's, pulmonary hemorrhage (PH) occurred mainly in full term infants with severe pre-existing illness. The incidence of PH was quoted as 1.3 per 1,000 live births. In the older medical literature, the risk factors associated with PH included the severity of the associated illness, intrauterine growth restriction, patent ductus arteriosus (PDA), coagulopathy, and the need for assisted ventilation. Presently, PH occurs mainly in preterm ventilated infants with severe respiratory distress syndrome (RDS) who often have a PDA and have received surfactant. Currently, PH complicates the hospital course of 3-5% of preterm infants with RDS. Although not clear, the cause of PH is thought to be due to a rapid lowering of intrapulmonary pressure, which facilitates left to right shunting across a patent ductus arteriosus and an increase in pulmonary blood flow. Retrospective case reports and one prospective uncontrolled study that used surfactant for PH in neonates have shown promising results in treating PH. OBJECTIVES: To evaluate the effect of surfactant treatment compared to placebo or no intervention on mortality in neonates with pulmonary hemorrhage. In addition, the review will evaluate the effect of surfactant treatment on neonatal morbidities associated with PH compared to placebo or no intervention. SEARCH STRATEGY: The following databases were searched in January 2008: The Cochrane Central Register of Controlled Trials (CENTRAL, The Cochrane Library, Issue 4, 2007) and MEDLINE from 1966 and EMBASE from 1980 to the time of the full review using the OVID interface. The proceedings of the Annual Meetings of the Pediatric Academic Societies and the European Society of Pediatric Research published in Pediatric Research or electronically on their web sites were searched from 1994 to the time of the full review. Science citation index (Web of Science) was searched for authors quoting key references of observational studies. SELECTION CRITERIA: Randomised or quasi randomised controlled trials that evaluated the effect of surfactant in the treatment of PH in intubated term or preterm (< 37 weeks) neonates with PH. Infants were included up to 44 weeks postmenstrual age. Infants were included regardless of prior treatment with surfactant. The interventions studied were intratracheal instillation of surfactant (natural or synthetic, regardless of dose) vs. placebo or no intervention. DATA COLLECTION AND ANALYSIS: If studies were identified by the literature search, the planned analyses included calculation of the relative risk (RR), risk difference (RD), number needed to treat (NNT) or number needed to harm (NNH) for dichotomous outcomes, and weighed mean difference (WMD) for continuous outcomes, with their 95% confidence intervals (CI). A fixed effects model would be used for meta-analyses. Heterogeneity tests, including the I- squared (I(2)) statistic, would be performed to assess the appropriateness of pooling the data and the results would be reported. MAIN RESULTS: No trials were identified. AUTHORS' CONCLUSIONS: No randomized or quasi-randomized trials that evaluated the effect of surfactant in PH were identified. Therefore, no conclusions from such trials can be drawn. In view of the promising results from studies with less strict study designs than a randomized controlled trial, there is reason to conduct further trials of surfactant for the treatment of PH in neonates. PMID- 18425915 TI - Recombinant interferon beta or glatiramer acetate for delaying conversion of the first demyelinating event to multiple sclerosis. AB - BACKGROUND: Immunomodulatory drugs have been shown to be only modestly effective in clinically definite relapsing remitting multiple sclerosis (RRMS). It has been hypothesized that their efficacy could be higher if used at the first appearance of symptoms, that is in the clinically isolated syndromes (CIS) suggestive of demyelinating events, a pathology which carries a high risk to convert to clinically definite MS (CDMS). OBJECTIVES: The objective of this review was to assess the effects of immunomodulatory drugs compared to placebo in adults in preventing conversion from CIS to CDMS which means the prevention of a second attack. SEARCH STRATEGY: We searched the Cochrane MS Group Trials Register (June 2007), Cochrane Central Register of Controlled Trials (CENTRAL)The Cochrane Library Issue 3, 2007, MEDLINE (January 1966 to June 2007), EMBASE (January 1974 to June 2007) and reference lists of articles. We also contacted manufacturers and researchers in the field. SELECTION CRITERIA: The trials selected were double blind, placebo-controlled, randomised trials of CIS patients treated with immunomodulatory drugs. DATA COLLECTION AND ANALYSIS: Study selection have been independently done by two reviewers. Two further reviewers independently assessed trial quality and extracted and analysed data. Study authors were contacted for additional informations. Adverse effects information was collected from the trials. MAIN RESULTS: Only three trials tested the efficacy of interferon (IFN) beta including a total of 1160 participants (639 treatment, 521 placebo); no trial tested the efficacy of glatiramer acetate (GA). The metanalyses showed that the proportion of patients converting to CDMS was significantly lower in IFN beta treated than in placebo-treated patients both after one year (pooled OR 0.53; 95% CI, 0.40 to 0.71; p <0.0001) as well as after two years of follow-up (pooled OR 0.52; 95% CI, 0.38 to 0.70; p <0.0001). Early treatment with IFN beta was associated with the side effect profile reported by the randomised controlled trials with this drug. Since side effects were reported with some heterogeneity in the three studies the metanalysis was possible only for the frequency of serious adverse events, not significantly different in IFN beta-treated or placebo-treated patients. AUTHORS' CONCLUSIONS: The efficacy of IFN beta treatment on preventing the conversion from CIS to CDMS was confirmed over two years of follow-up. Since patients had some clinical heterogeneity (length of follow-up, clinical findings of initial attack), it could be useful for the clinical practice to further analyse the efficacy of IFN beta treatment in different patient subgroups. PMID- 18425916 TI - Chinese herbal medicine for primary dysmenorrhoea. AB - BACKGROUND: Conventional treatment for primary dysmenorrhoea has a failure rate of 20% to 25% and may be contraindicated or not tolerated by some women. Chinese herbal medicine may be a suitable alternative. OBJECTIVES: To determine the efficacy and safety of Chinese herbal medicine for primary dysmenorrhoea when compared with placebo, no treatment, and other treatment. SEARCH STRATEGY: The Cochrane Menstrual Disorders and Subfertility Group Trials Register (to 2006), MEDLINE (1950 to January 2007), EMBASE (1980 to January 2007), CINAHL (1982 to January 2007), AMED (1985 to January 2007), CENTRAL (The Cochrane Library issue 4, 2006), China National Knowledge Infrastructure (CNKI, 1990 to January 2007), Traditional Chinese Medicine Database System (TCMDS, 1990 to December 2006), and the Chinese BioMedicine Database (CBM, 1990 to December 2006) were searched. Citation lists of included trials were also reviewed. SELECTION CRITERIA: Any randomised controlled trials involving Chinese herbal medicine versus placebo, no treatment, conventional therapy, heat compression, another type of Chinese herbal medicine, acupuncture or massage. Exclusion criteria were identifiable pelvic pathology and dysmenorrhoea resulting from the use of an intra-uterine contraceptive device. DATA COLLECTION AND ANALYSIS: Quality assessment, data extraction and data translation were performed independently by two review authors. Attempts were made to contact study authors for additional information and data. Data were combined for meta-analysis using either Peto odds ratios or relative risk (RR) for dichotomous data or weighted mean difference for continuous data. A fixed-effect statistical model was used, where suitable. If data were not suitable for meta-analysis, any available data from the trial were extracted and presented as descriptive data. MAIN RESULTS: Thirty-nine randomised controlled trials involving a total of 3475 women were included in the review. A number of the trials were of small sample size and poor methodological quality. Results for Chinese herbal medicine compared to placebo were unclear as data could not be combined (3 RCTs). Chinese herbal medicine resulted in significant improvements in pain relief (14 RCTs; RR 1.99, 95% CI 1.52 to 2.60), overall symptoms (6 RCTs; RR 2.17, 95% CI 1.73 to 2.73) and use of additional medication (2 RCTs; RR 1.58, 95% CI 1.30 to 1.93) when compared to use of pharmaceutical drugs. Self-designed Chinese herbal formulae resulted in significant improvements in pain relief (18 RCTs; RR 2.06, 95% CI 1.80 to 2.36), overall symptoms (14 RCTs; RR 1.99, 95% CI 1.65 to 2.40) and use of additional medication (5 RCTs; RR 1.58, 95% CI 1.34 to 1.87) after up to three months of follow-up when compared to commonly used Chinese herbal health products. Chinese herbal medicine also resulted in better pain relief than acupuncture (2 RCTs; RR 1.75, 95% CI 1.09 to 2.82) and heat compression (1 RCT; RR 2.08, 95% CI 2.06 to 499.18). AUTHORS' CONCLUSIONS: The review found promising evidence supporting the use of Chinese herbal medicine for primary dysmenorrhoea; however, results are limited by the poor methodological quality of the included trials. PMID- 18425918 TI - Physical activity and enhanced fitness to improve cognitive function in older people without known cognitive impairment. AB - BACKGROUND: Physical activity is beneficial for healthy ageing. It may also help maintain good cognitive function in older age. Aerobic activity improves cardiovascular fitness, but it is not known whether this sort of fitness is necessary for improved cognitive function. Studies in which activity, fitness and cognition are reported in the same individuals could help to resolve this question. OBJECTIVES: To assess the effectiveness of physical activity, aimed at improving cardiorespiratory fitness, on cognitive function in older people without known cognitive impairment. SEARCH STRATEGY: We searched MEDLINE, EMBASE, PEDro, SPORTDiscus, PsycINFO, CINAHL, Cochrane Controlled Trials Register (CENTRAL), Dissertation abstracts international and ongoing trials registers on 15 December 2005 with no language restrictions. SELECTION CRITERIA: All published randomised controlled trials comparing aerobic physical activity programmes with any other intervention or no intervention with participants older than 55 years of age were eligible for inclusion. DATA COLLECTION AND ANALYSIS: Eleven RCTs fulfilling the inclusion criteria are included in this review. Two reviewers independently extracted the data from these included studies. MAIN RESULTS: Eight out of 11 studies reported that aerobic exercise interventions resulted in increased cardiorespiratory fitness of the intervention group (an improvement on the maximum oxygen uptake test which is considered to be the single best indicator of the cardiorespiratory system) of approximately 14% and this improvement coincided with improvements in cognitive capacity. The largest effects on cognitive function were found on motor function, auditory attention and delayed memory functions (effect sizes of 1.17, 0.52 and 0.50 respectively). However, the results for delayed memory functions should be interpreted with care since they are based on a single study. Moderate effects were observed for cognitive speed (speed at which information is processed; effect size 0.26) and visual attention (effect size 0.26). AUTHORS' CONCLUSIONS: There is evidence that aerobic physical activities which improve cardiorespiratory fitness are beneficial for cognitive function in healthy older adults, with effects observed for motor function, cognitive speed, delayed memory functions and auditory and visual attention. However, the majority of comparisons yielded no significant results. The data are insufficient to show that the improvements in cognitive function which can be attributed to physical exercise are due to improvements in cardiovascular fitness, although the temporal association suggests that this might be the case. Larger studies are still required to confirm whether the aerobic training component is necessary, or whether the same can be achieved with any type of physical exercise. At the same time, it would be informative to understand why some cognitive functions seem to improve with (aerobic) physical exercise while other functions seem to be insensitive to physical exercise. Clinicians and scientists in the field of neuropsychology should seek mutual agreement on a smaller battery of cognitive tests to use, in order to render research on cognition clinically relevant and transparent and heighten the reproducibility of results for future research. PMID- 18425917 TI - Monitoring of stimulated cycles in assisted reproduction (IVF and ICSI). AB - BACKGROUND: Traditional monitoring of ovarian hyperstimulation during in vitro fertilisation (IVF) treatment has included ultrasonography plus serum estradiol concentration to ensure safe practice by reducing the incidence and severity of ovarian hyperstimulation syndrome (OHSS). The need for intensive monitoring during ovarian stimulation in IVF is controversial. It has been suggested that close monitoring is time consuming, expensive and inconvenient for the woman and simplification of IVF therapy by using ultrasound only should be considered. This systematic review assessed the effects of ovarian monitoring by ultrasound only versus ultrasound plus serum estradiol measurement on IVF outcomes and the occurrence of OHSS in women undergoing stimulated cycles in IVF and intra cytoplasmic sperm injection (ICSI) treatment. OBJECTIVES: To quantify the effect of monitoring controlled ovarian stimulation in IVF and ICSI cycles with ultrasound plus serum estradiol concentration versus ultrasound only in terms of live birth rates, pregnancy rates and the incidence of OHSS. SEARCH STRATEGY: We searched the Menstrual Disorders and Subfertility Group Specialised Register of controlled trials, Cochrane Central Register of Controlled Trials (CENTRAL) on the latest issue of The Cochrane Library, MEDLINE (1966 to May 2007), EMBASE (1980 to May 2007), CINAHL (1982 to May 2007), the National Research Register, and web-based trial databases such as Current Controlled Trials. There was no language restriction. Additionally all references in the identified trials and background papers were checked and authors were contacted to identify relevant published and unpublished data. SELECTION CRITERIA: Only randomised controlled trials that compared monitoring with ultrasound plus serum estradiol concentration versus ultrasound only in women undergoing ovarian hyperstimulation for IVF and ICSI treatment were included. DATA COLLECTION AND ANALYSIS: Two review authors independently examined the electronic search results for relevant trials, extracted data and assessed trial quality. They resolved disagreements by discussion with two other authors. Outcomes data were pooled when appropriate and summary statistics presented when limited data did not allow meta-analysis. MAIN RESULTS: Our search strategy identified 1119 potentially eligible reports, of which two met our inclusion criteria. These involved 411 women who underwent controlled ovarian stimulation monitoring. Our primary outcome of live birth rate was not reported in either study. One trial reported clinical pregnancy rate per woman (33% versus 31%; RR 1.07, 95% CI 0.77 to 1.49), the second trial reported clinical pregnancy rate per oocyte retrieval (22% versus 25%). There was no significant difference between the ultrasound plus estradiol group and the ultrasound alone group in the mean number of oocytes retrieved (WMD -0.55, 95% CI -1.79 to 0.69) and the incidence of ovarian hyperstimulation (RR 0.73, 95% CI 0.30 to 1.78) for the two studies. AUTHORS' CONCLUSIONS: There is no evidence from randomised trials to support cycle monitoring by ultrasound plus serum estradiol as more efficacious than cycle monitoring by ultrasound only on outcomes of live birth and pregnancy rates. A large well-designed randomised controlled trial is needed that reports on live birth rates and pregnancy, with economic evaluation of the costs involved and the views of the women undergoing cycle monitoring. A randomised trial with sufficiently large sample size to test the effects of different monitoring protocols on OHSS, a rare outcome, will pose a great challenge. Until such a trial is considered feasible, cycle monitoring by transvaginal ultrasound plus serum estradiol may need to be retained as a precautionary good practice point. PMID- 18425919 TI - Antacids for preventing oesophagogastric variceal bleeding and rebleeding in cirrhotic patients. AB - BACKGROUND: Ruptured gastroesophageal varices are the most severe and frequent cause of gastrointestinal bleeding in cirrhotic patients, leading to death in 5% to 8% of patients during the first 48 hours and oesophagogastric varices account for 60% to 80% of first bleeding in patients with portal hypertension. Antacids are often used for emergency treatment of bleeding oesophageal varices in patients with cirrhosis of the liver. OBJECTIVES: To evaluate the beneficial and harmful effects of antacids for preventing oesophagogastric bleeding and rebleeding. SEARCH STRATEGY: We identified relevant randomised clinical trials by searching The Cochrane Hepato-Biliary Group Controlled Trials Register (June 2007), the Cochrane Central Register of Controlled Trials (CENTRAL) in The Cochrane Library (Issue 2, 2007), MEDLINE (1950 to June 2007), EMBASE (Excerpta Medica Database) (1980 to June 2007), and the Science Citation Index Expanded (SCI-EXPANDED) (1945 to June 2007). Additional randomised trials were sought from the reference lists of the trials found and reviews identified by the electronic searches. SELECTION CRITERIA: We planned to include randomised clinical trials. DATA COLLECTION AND ANALYSIS: We planned to summarise data using Cochrane Collaboration methodologies. MAIN RESULTS: We could not find any randomised clinical trials on antacids for preventing oesophagogastric variceal bleeding and bleeding in cirrhotic patients. AUTHORS' CONCLUSIONS: it is not possible to determine whether antacids are beneficially effective or harmful for preventing oesophagogastric variceal bleeding and rebleeding in cirrhotic patients since randomised clinical trials investigating it are lacking. PMID- 18425920 TI - Systemic corticosteroids for acute gout. AB - BACKGROUND: Gout is one of the most frequently occurring rheumatic diseases, worldwide. Given the well-known drawbacks of the regular treatments for acute gout (non-steroidal anti-inflammatory drugs (NSAIDs), colchicine), systemic corticosteroids might be safe alternatives. OBJECTIVES: To assess the efficacy and safety of systemic corticosteroids in the treatment of acute gout in comparison with placebo, NSAIDs, colchicine, other active drugs, other therapies, or no therapy. SEARCH STRATEGY: Searches were done in the following electronic databases: Cochrane Central Register of Controlled Trials (CENTRAL) (The Cochrane Library 2007); MEDLINE (1966 to 2007) through PubMed; EMBASE (1974 to 2007); Web of Science (1975 to 2007); LILACS (1986 to 2007); and databases of ongoing trials (up to April 2007). SELECTION CRITERIA: Randomized controlled trials and controlled clinical trials investigating the use of systemic corticosteroids in the treatment of acute gout were included. DATA COLLECTION AND ANALYSIS: Two review authors decided independently which trials to include. The same review authors also collected the data in a standardised form and assessed the methodological quality of the trial using validated criteria. When possible, continuous and dichotomous data were summarised statistically. MAIN RESULTS: Three head to head trials involving 148 patients (74 systemic corticosteroids; 74 comparator drugs) were included. Placebo-controlled trials were not found. In the studies, different kinds of systemic corticosteroids and different kinds of control drugs were used, both administered in different routes. Intramuscular triamcinolone acetonide was compared respectively to oral indomethacine, and intramuscular adrenocorticotropic hormone (ACTH); oral prednisolone (together with a single intramuscular diclophenac injection) was compared to oral indomethacine (together with a single placebo injection). Outcome measurements varied: average number of days until total relief of signs, mean decrease of pain per unit of time in mm on a visual analogue scale (VAS) - during rest and activity. In the triamcinolone-indomethacine trial the clinical joint status was used as an additional outcome. Clinically relevant differences between the studied systemic corticosteroids and the comparator drugs were not found; important safety problems attributable to the used corticosteroids were not reported. The quality of the three studies was graded as very low to moderate. Statistical pooling of results was not possible. AUTHORS' CONCLUSIONS: There is inconclusive evidence for the efficacy and effectiveness of systemic corticosteroids in the treatment of acute gout. Patients with gout did not report serious adverse effects from systemic corticosteroids, when used short term. PMID- 18425921 TI - Commercial versus home-made spacers in delivering bronchodilator therapy for acute therapy in children. AB - BACKGROUND: Strong evidence supports the use of metered-dose inhalers combined with a spacer for delivering rapid-acting inhaled beta-2 agonists in the treatment of acute exacerbations of asthma in children. The high cost and lack of availability of commercially produced spacers however, have limited their use in developing countries. OBJECTIVES: The aim of this review was to compare the response to inhaled beta-2 agonists delivered through metered-dose inhaler using home-made spacers, to the use of commercially produced spacers, in children with acute exacerbations of wheezing or asthma. SEARCH STRATEGY: We searched the Cochrane Central Register of Controlled Trials (CENTRAL) The Cochrane Library Issue 3, 2007,(up to August 2007) MEDLINE , EMBASE, CINHAL, LILACS and reference lists of included studies. We contacted authors and known experts in the field, and approached pharmaceutical companies that manufacture inhalation spacers to identify additional published or unpublished data. No language restrictions were applied. SELECTION CRITERIA: Trials comparing treatment with rapid acting beta 2 agonists delivered though MDI attached to home-made spacers, with the same bronchodilator therapy delivered with MDI and commercially produced spacers, in children under 18 years with acute exacerbations of wheezing or asthma. DATA COLLECTION AND ANALYSIS: Two review authors independently extracted the data and assessed trial quality. Missing data were obtained from the authors or estimated from information available in published reports. MAIN RESULTS: Six trials with 658 participants met the inclusion criteria . At the time of this report, five trials were published in full text, and one study was available in abstract form only. No significant differences were demonstrated between the two delivery methods in terms of need for hospital admission (RR 1.00, 95% CI 0.63 to 1.59), change in oxygen saturation (SMD -0.03, 95% CI -0.39 to 0.33), PEFR (SMD 0.04, 95% CI -0.72 to 0.80), clinical score (WMD 0.00, 95% CI -0.37 to 0.37), in terms of need for additional treatment (RR 1.18, 95% CI 0.84 to 1.65), or regarding change in heart rate per minute (SMD 0.09, 95% CI -0.24 to 0.42). AUTHORS' CONCLUSIONS: Overall, this review supports did not identify a difference between these two methods for delivering bronchodilator therapy to children with acute asthma or lower airways obstruction attacks. Care should be taken in the interpretation and applicability of our results because of the small number of RCTs along with few events available meeting the criteria for inclusion in the review, absence of the primary outcome of interest and other clinically important outcomes in the majority of included studies. The possible need for a face-mask in younger children using home-made spacers should also be considered in practice. PMID- 18425922 TI - Psychotherapy for depression among incurable cancer patients. AB - BACKGROUND: The most common psychiatric diagnosis among cancer patients is depression; this diagnosis is even more common among patients with advanced cancer. Psychotherapy is a patient-preferred and promising strategy for treating depression among cancer patients. Several systematic reviews have investigated the effectiveness of psychological treatment for depression among cancer patients. However, the findings are conflicting, and no review has focused on depression among patients with incurable cancer. OBJECTIVES: To investigate the effects of psychotherapy for treating depression among patients with advanced cancer by conducting a systematic review of randomized controlled trials (RCTs). SEARCH STRATEGY: We searched the Cochrane Pain, Palliative and Supportive Care Group Register, The Cochrane Controlled Trials Register, MEDLINE, EMBASE, CINAHL, and PsycINFO databases in September 2005. SELECTION CRITERIA: All relevant RCTs comparing any kind of psychotherapy with conventional treatment for adult patients with advanced cancer were eligible for inclusion. Two independent review authors identified relevant studies. DATA COLLECTION AND ANALYSIS: Two review authors independently extracted data from the original reports using standardized data extraction forms. Two independent review authors also assessed the methodological quality of the selected studies according to the recommendations of a previous systematic review of psychological therapies for cancer patients that utilized ten internal validity indicators. The primary outcome was the standardized mean difference (SMD) of change between the baseline and immediate post-treatment scores. MAIN RESULTS: We identified a total of ten RCTs (total of 780 participants); data from six studies were used for meta-analyses (292 patients in the psychotherapy arm and 225 patients in the control arm). Among these six studies, four studies used supportive psychotherapy, one adopted cognitive behavioural therapy, and one adopted problem-solving therapy. When compared with treatment as usual, psychotherapy was associated with a significant decrease in depression score (SMD = -0.44, 95% confidence interval [CI] = -0.08 to -0.80). None of the studies focused on patients with clinically diagnosed depression. AUTHORS' CONCLUSIONS: Evidence from RCTs of moderate quality suggest that psychotherapy is useful for treating depressive states in advanced cancer patients. However, no evidence supports the effectiveness of psychotherapy for patients with clinically diagnosed depression. PMID- 18425923 TI - Topotecan for ovarian cancer. AB - BACKGROUND: Chemotherapeutic agents such as topotecan can be used to treat ovarian cancer. The effects of using topotecan as a therapeutic agent have not been previously been systematically reviewed. OBJECTIVES: To systematically evaluate the effectiveness and safety of topotecan for the treatment of ovarian cancer. SEARCH STRATEGY: We searched the Cochrane Central Register of Controlled Trials (CENTRAL), (Issue 4, 2006); Cochrane Gynaecological Cancer Review Group (CGCRG) Specialised Register (Cochrane Library Issue 4, 2006); MEDLINE (January 1990 to 27 July 2006); EMBASE (January 1990 to 27 July 2006); The European Organization for the Research and Treatment of Cancer (EORTC) database (to 1 August 2006); CBM (Chinese Biomedical Database) (January 1990 to 27 July 2006). SELECTION CRITERIA: Randomised controlled trials (RCTs) which randomized patients with ovarian cancer to single or combined use of topotecan versus interventions without topotecan, or different remedies of topotecan. DATA COLLECTION AND ANALYSIS: Two review authors independently extracted and analysed data. MAIN RESULTS: Six studies including 1323 participants were eligible for this review (Gordon 2004a; Gore 2001a; Gore 2002; Hoskins 1998; Huinink 2004; Placido 2004) All studies, as reported, were identified as being of poor methodological quality. Topotecan had comparable effectiveness to prolong progression-free survival (PFS) compared with pegylated liposomal doxorubicin (PLD), (16.1 weeks versus 17.0 weeks; p = 0.095). Overall survival (OS) time was similar in participants using PLD compared with topotecan (56.7 weeks versus 60 weeks; p = 0.341). Topotecan was more hematologically toxic compared with paclitaxel or PLD, relative risks (RRs) of hematological events: ranged from 1.03 to 14.46 and 1.73 to 27.12 respectively. A 21-day cycle of topotecan was more toxic than a 42-day cycle (RRs of hematological and non-hematological events ranged from 1.03 to 8). Intravenous and oral topotecan had comparable toxicity. Topotecan delayed progression more effectively compared with paclitaxel (23.1 weeks versus 14 weeks, p = 0.0021). Participants were more likely to respond to topotecan on a 21 day cycle as opposed to a 42-day cycle (RR 7.23, 95% CI 0.94 to 55.36). Small tumor diameter, sensitivity to platinum-based chemotherapy was associated with better prognosis. Small sample size, methodological flaws and poor reporting of the included trials made measurement bias of the trials difficult to assess. AUTHORS' CONCLUSIONS: Topotecan appears to have a similar level of effectiveness as paclitaxel and PLD, though with different patterns of side effects. Larger, well-designed RCTs are required in order to define an optimal regime. PMID- 18425924 TI - Huperzine A for Alzheimer's disease. AB - BACKGROUND: Alzheimer's disease (AD) has become a major public health problem around the world due to its increasing prevalence, long duration, caregiver burden, and high financial cost of care. The degeneration of acetylcholine containing neurons in the basal forebrain has been implicated in the symptoms of AD. Cholinesterase inhibitors may block the degradation of acetylcholine, thus increasing the efficacy of the remaining cholinergic neurons. Huperzine A is a linearly competitive, reversible inhibitor of acetyl cholinesterase that is said to have both central and peripheral activity with the ability to protect cells against hydrogen peroxide, beta-amyloid protein (or peptide), glutamate, ischemia and staurosporine-induced cytotoxicity and apoptosis. These properties might qualify Huperzine A as a promising agent for treating dementia (including AD). OBJECTIVES: To assess the efficacy and safety of Huperzine A for the treatment of patients with AD. SEARCH STRATEGY: The Specialized Register of the Cochrane Dementia and Cognitive Improvement Group was searched on 1 February 2006 using the search term: huperzin*. The CDCIG Specialized register contains records from all major health care databases (MEDLINE, EMBASE, PsycINFO, CINAHL, SIGLE, ISTP, INSIDE, LILACS) as well as from many trials databases and grey literature sources. In addition, the CBM and AMED databases and relevant websites were searched and some journals were hand-searched. Specialists in the field were approached for unpublished material and any publications found were searched for additional references. SELECTION CRITERIA: All relevant randomized controlled trials (RCTs) studying the efficacy and safety of Huperzine A for AD. DATA COLLECTION AND ANALYSIS: Data were extracted independently by two reviewers using a self-developed data extraction form and entered into RevMan 4.2.10 software. Meta-analyses were performed when more than one trial provided data on a comparable outcome on sufficiently similar patients. Random effects analyses were performed whenever heterogeneity between results appeared to be present. Standardized differences in mean outcome measures were used due to the use of different scales and periods of treatment. MAIN RESULTS: Six trials including a total of 454 patients met our inclusion criteria. The methodological quality of most included trials was not high. It was shown that compared to placebo, Huperzine A had beneficial effects on the improvement of general cognitive function measured by MMSE (WMD 2.81; 95% CI 1.87 to 3.76; P < 0.00001) and ADAS Cog at six weeks (WMD 1.91; 95% CI 1.27 to 2.55) and at 12 weeks (WMD 2.51; 95% CI 1.74 to 3.28), global clinical assessment measured by CDR (WMD -0.80; 95% CI 0.95 to -0.65) and CIBIC-plus (OR 4.32, 95% CI 2.37 to 7.90), behavioral disturbance measured by ADAS-non-Cog at six weeks (WMD -1.33, 95%CI -2.12 to 0.54) and at 12 weeks (WMD -1.52, 95% CI-2.39 to -0.65), and functional performance measured by ADL (WMD = -7.17; 95% CI -9.13 to -5.22; P < 0.00001). However, Huperzine A was not superior to placebo in the improvement of general cognitive function measured by Hasegawa Dementia Scale (HDS) (WMD: 2.78; 95% CI 0.17 to 5.73, P = 0.06) and specific cognitive function measured by Weshler Memory Scale (WMS) (WMD = 6.64; 95% CI -3.22 to 16.50; P = 0.19). No data were available on quality of life and caregiver burden. The adverse events of Huperzine A were mild and there were no significant differences of adverse events between Huperzine A groups and control groups. AUTHORS' CONCLUSIONS: From the available evidence, Huperzine A seems to have some beneficial effects on improvement of general cognitive function, global clinical status, behavioral disturbance and functional performance, with no obvious serious adverse events for patients with AD. However, only one study was of adequate quality and size. There is therefore inadequate evidence to make any recommendation about its use. Rigorous design, randomized, multi-centre, large-sample trials of Huperzine A for AD are needed to further assess the effects. PMID- 18425925 TI - Anti-histamines for prolonged non-specific cough in children. AB - BACKGROUND: Non-specific cough is defined as non-productive cough in the absence of identifiable respiratory disease or known aetiology. It is commonly seen in paediatric practice. These children are treated with a variety of therapies including anti-histamines. Also, anti-histamines are advocated as an empirical treatment in adults with chronic cough. OBJECTIVES: To evaluate the effectiveness of anti-histamines in treating children with prolonged non-specific cough. SEARCH STRATEGY: We searched the Cochrane Register of Controlled Trials (CENTRAL), MEDLINE, OLDMEDLINE and EMBASE databases. The latest searches were performed in November 2007. SELECTION CRITERIA: All randomised controlled trials comparing anti-histamines with a placebo or placebo-like medication with cough as an outcome, where cough is not primarily related to an underlying respiratory disorder such as cystic fibrosis, asthma, or suppurative lung disease. DATA COLLECTION AND ANALYSIS: Two review authors independently assessed study quality and extracted data. MAIN RESULTS: Three included therapeutic studies had 182 randomised participants with 162 completing the trials although in one study, children with recurrent wheeze were also included. The two included safety evaluation studies randomised 963 participants with 793 completing the trials. Clinical heterogeneity was evident and limited data prevented combining data for meta-analysis. The two larger therapeutic studies described significant improvement in both the intervention and the placebo/placebo-like arms with no significant difference between the two groups. In the study with the smallest sample size, cetirizine (a second generation anti-histamine) was significantly more efficacious than placebo in reducing chronic cough in children associated with seasonal allergic rhinitis, and the effect was seen within two weeks of therapy. Combined data from the safety evaluation studies revealed a non significant difference between groups (OR 1.6, 95% CI 0.7 to 3.82) for cough as an adverse event but the trend favoured the placebo arm. AUTHORS' CONCLUSIONS: This review has significant limitations. However, our finding of uncertain efficacy of anti-histamines for chronic cough are similar to that for acute cough in children. In contrast to recommendations in adults with chronic cough, anti histamines cannot be recommended as empirical therapy for children with chronic cough. If anti-histamines were to be trialled in these children, current data suggest a clinical response (time to response) occurs within two weeks of therapy. However the use of anti-histamines in children with non-specific cough has to be balanced against the well known risk of adverse events especially in very young children. PMID- 18425926 TI - Antibiotics to reduce post-tonsillectomy morbidity. AB - BACKGROUND: Tonsillectomy continues to be one of the most common surgical procedures performed in children and adults. Despite improvements in surgical and anaesthetic techniques, postoperative morbidity, mainly in the form of pain, remains a significant clinical problem. Postoperative bacterial infection of the tonsillar fossa has been proposed as an important factor causing pain and associated morbidity, and some studies have found a reduction in morbid outcomes following the administration of perioperative antibiotics. OBJECTIVES: To determine whether perioperative antibiotics reduce pain and other morbid outcomes following tonsillectomy. SEARCH STRATEGY: Cochrane ENT Group Trials Register, Cochrane Central Register of Controlled Trials (CENTRAL) (The Cochrane Library, Issue 1 2007), MEDLINE (1950 to 2007) and EMBASE (1974 to 2007) were searched. The date of the last search was March 2007. SELECTION CRITERIA: All randomised controlled trials examining the impact of perioperative administration of systemic antibiotics on post-tonsillectomy morbidity in children or adults. DATA COLLECTION AND ANALYSIS: Two authors independently collected data. Primary outcomes were pain, consumption of analgesia and secondary haemorrhage (defined as significant if patient re-admitted, transfused blood products or returned to theatre, and total if any documented haemorrhage). Secondary outcomes were fever, time taken to resume normal diet and activities and adverse events. Where possible, summary measures were generated using random-effects models. MAIN RESULTS: Nine trials met the eligibility criteria. Most did not find a significant reduction in pain with antibiotics. Similarly, antibiotics were not shown to be effective in reducing the need for analgesics. Antibiotics were not associated with a reduction in significant secondary haemorrhage rates (Relative Risk (RR) 0.49, 95% CI 0.08 to 3.11, P = 0.45) or total secondary haemorrhage rates (RR 0.92, 95% CI 0.45 to 1.87, P = 0.81). With regard to secondary outcomes, antibiotics reduced the proportion of subjects with fever (RR 0.63, 95% CI 0.46 to 0.85, P = 0.002). AUTHORS' CONCLUSIONS: The present review suggests that there is little or no evidence that antibiotics reduce the main morbid outcomes following tonsillectomy (i.e. pain, the need for analgesia or secondary haemorrhage rates). They do however appear to reduce fever. Some important methodological shortcomings exist in the included trials which are likely to have produced bias favouring antibiotics. We therefore advocate caution when prescribing antibiotics routinely to all patients undergoing tonsillectomy. Whether a subgroup of patients who might benefit from selective administration of antibiotics exists is unknown and needs to be explored in future trials. PMID- 18425927 TI - Non-pharmacological interventions for breathlessness in advanced stages of malignant and non-malignant diseases. AB - BACKGROUND: Breathlessness is a common and distressing symptom in the advanced stages of malignant and non-malignant diseases. Appropriate management requires both pharmacological and non-pharmacological interventions. OBJECTIVES: The primary objective was to determine the effectiveness of non-pharmacological and non-invasive interventions to relieve breathlessness in participants suffering from the five most common conditions causing breathlessness in advanced disease. SEARCH STRATEGY: We searched the following databases: The Cochrane Central Register of Controlled Trials, MEDLINE, EMBASE, CINAHL, British Nursing Index, PsycINFO, Science Citation Index Expanded, AMED, The Cochrane Pain, Palliative and Supportive Care Trials Register, The Cochrane Database of Systematic Reviews, and Database of Abstracts of Reviews of Effectiveness in June 2007. We also searched various websites and reference lists of relevant articles and textbooks. SELECTION CRITERIA: We included randomised controlled and controlled clinical trials assessing the effects of non-pharmacological and non-invasive interventions to relieve breathlessness in participants described as suffering from breathlessness due to advanced stages of cancer, chronic obstructive pulmonary disease (COPD), interstitial lung disease, chronic heart failure or motor neurone disease. DATA COLLECTION AND ANALYSIS: Two review authors independently assessed relevant studies for inclusion. Data extraction and quality assessment was performed by three review authors and checked by two other review authors. Meta-analysis was not attempted due to heterogeneity of studies. MAIN RESULTS: Forty-seven studies were included (2532 participants) and categorised as follows: single component interventions with subcategories of walking aids (n = 7), distractive auditory stimuli (music) (n = 6), chest wall vibration (CWV, n = 5), acupuncture/acupressure (n = 5), relaxation (n = 4), neuro-electrical muscle stimulation (NMES, n = 3) and fan (n = 2). Multi component interventions were categorised in to counselling and support (n = 5), breathing training (n = 3), counselling and support with breathing-relaxation training (n = 2), case management (n = 2) and psychotherapy (n = 2). There was a high strength of evidence that NMES and CWV could relieve breathlessness and moderate strength for the use of walking aids and breathing training. There is a low strength of evidence that acupuncture/acupressure is helpful. There is not enough data to judge the evidence for distractive auditory stimuli (music), relaxation, fan, counselling and support, counselling and support with breathing relaxation training, case management and psychotherapy. Most studies have been conducted in COPD patients, only a few studies included participants with other conditions. AUTHORS' CONCLUSIONS: Breathing training, walking aids, NMES and CWV appear to be effective non-pharmacological interventions for relieving breathlessness in advanced stages of disease. PMID- 18425928 TI - Therapy-based rehabilitation services for patients living at home more than one year after stroke. AB - BACKGROUND: Current practice of rehabilitation intervention mainly concentrates on the first six months of stroke. At present, there is no agreed consensus about the benefits of such a service more than one year after stroke. OBJECTIVES: To ascertain whether therapy-based rehabilitation services can influence outcome one year or more after stroke. SEARCH STRATEGY: We searched the trials registers of the following Cochrane Review Groups: Stroke Group (last searched September 2007), Effective Practice and Organisation of Care Group (last searched October 2006) and Dementia and Cognitive Improvement Group (last searched October 2006). We also searched the Cochrane Central Register of Controlled Trials (CENTRAL) (The Cochrane Library Issue 4, 2006), MEDLINE (1966 to October 2006), EMBASE (1980 to October 2006), CINAHL (1982 to October 2006), AMED (1985 to October 2006), PEDro (1952 to October 2006), British Nursing Index (1993 to October 2006), DARE (1994 to October 2006), HMIC (1979 to October 2006) and NHS EED (1991 to October 2006). We also searched dissertation databases and ongoing trials and research registers, scanned reference lists and contacted researchers and experts in the field. SELECTION CRITERIA: All randomised controlled trials of community based stroke patients, in which at least 75% were recruited one year after stroke and received a therapy-based rehabilitation intervention that was compared with conventional care. DATA COLLECTION AND ANALYSIS: Two review authors independently selected trials and extracted data on a number of pre-specified outcomes. The primary outcomes were the proportion of participants who had deteriorated or were dependent in personal activities of daily living at the end of scheduled follow up. MAIN RESULTS: We identified five trials of 487 participants that were eligible for the review. Overall, there was inconclusive evidence as to whether therapy-based rehabilitation intervention one year after stroke was able to influence any relevant patient or carer outcome. Trials varied in design, type of interventions provided, quality, and outcomes assessed. AUTHORS' CONCLUSIONS: This review highlights the dearth of evidence investigating long-term therapy based rehabilitation interventions for patients with stroke. PMID- 18425929 TI - Exercise therapy in juvenile idiopathic arthritis. AB - BACKGROUND: Exercise therapy is considered an important component of the treatment of arthritis. The efficacy of exercise therapy has been reviewed in adults with rheumatoid arthritis but not in children with juvenile idiopathic arthritis (JIA). OBJECTIVES: To assess the effects of exercise therapy on functional ability, quality of life and aerobic capacity in children with JIA. SEARCH STRATEGY: The Cochrane Central Register of Controlled Trials (CENTRAL), Cochrane Database of Systematic Reviews (The Cochrane Library), MEDLINE (January 1966 to April 2007), CINAHL (January 1982 to April 2007), EMBASE (January 1966 to October 2007), PEDro (January 1966 to October 2007), SportDiscus (January 1966 to October 2007), Google Scholar (to October 2007), AMED (Allied and Alternative Medicine) (January 1985 to October 2007), Health Technologies Assessment database (January 1988 to October 2007), ISI Web Science Index to Scientific and Technical Proceedings (January 1966 to October 2007) and the Chartered Society of Physiotherapy website (http://www.cps.uk.org) were searched and references tracked. SELECTION CRITERIA: Randomised controlled trials (RCTs) of exercise treatment in JIA. DATA COLLECTION AND ANALYSIS: Potentially relevant references were evaluated and all data were extracted by two review authors working independently. MAIN RESULTS: Three out of 16 identified studies met the inclusion criteria, with a total of 212 participants. All the included studies fulfilled at least seven of 10 methodological criteria. The outcome data of the following measures were homogenous and were pooled in a meta-analysis: functional ability (n = 198; WMD -0.07, 95% CI -0.22 to 0.08), quality of life (CHQ-PhS: n = 115; WMD -3.96, 95% CI -8.91 to 1.00) and aerobic capacity (n = 124; WMD 0.04, 95% CI 0.11 to 0.19). The results suggest that the outcome measures all favoured the exercise therapy but none were statistically significant. None of the studies reported negative effects of the exercise therapy. AUTHORS' CONCLUSIONS: Overall, based on 'silver-level' evidence (www.cochranemsk.org) there was no clinically important or statistically significant evidence that exercise therapy can improve functional ability, quality of life, aerobic capacity or pain. The low number of available RCTs limits the generalisability. The included and excluded studies were all consistent about the adverse effects of exercise therapy; no short-term detrimental effects of exercise therapy were found in any study. Both included and excluded studies showed that exercise does not exacerbate arthritis. The large heterogeneity in outcome measures, as seen in this review, emphasises the need for a standardised assessment or a core set of functional and physical outcome measurements suited for health research to generate evidence about the possible benefits of exercise therapy for patients with JIA. Although the short term effects look promising, the long-term effect of exercise therapy remains unclear. PMID- 18425930 TI - Short-course versus long-course antibiotic therapy for non-severe community acquired pneumonia in children aged 2 months to 59 months. AB - BACKGROUND: Pneumonia is the leading cause of mortality in children under five years of age. Treatment of pneumonia requires an effective antibiotic used in adequate doses for an appropriate duration. Recommended duration of treatment ranges between 7 and 14 days, but this is not based on any empirical evidence. Shorter duration of therapy, if found to be effective, could be particularly important in resource-poor settings where there is a high risk of death, poor access to medicines and health care, and limited budgets for medicines. OBJECTIVES: To evaluate the efficacy of short-course versus long-course therapy with the same antibiotic for non-severe community-acquired pneumonia in children aged 2 to 59 months. SEARCH STRATEGY: We searched The Cochrane Central Register of Controlled Trials (CENTRAL); the Database of Abstracts of Reviews of Effects (DARE) (The Cochrane Library, 2007, Issue 3); MEDLINE (OVID) (January 1966 to September 2007); EMBASE (Embase.com) (1974 to September 2007); and LILACS (1982 to September 2007). SELECTION CRITERIA: All randomized controlled trials (RCTs) evaluating the efficacy of short-course versus long-course therapy using the same antibiotic for non-severe community-acquired pneumonia in children. DATA COLLECTION AND ANALYSIS: Two review authors independently assessed trial quality and extracted the data. MAIN RESULTS: Three studies (5763 children) were included. Analysis of three days versus five days of treatment with the same antibiotic for non-severe pneumonia in children showed non-significant differences in rates of clinical cure at the end of treatment (RR 0.99; 95% CI 0.97 to 1.01), treatment failure at the end of treatment (RR 1.07; 95% CI 0.92 to 1.25) and relapse rate after seven days of clinical cure (RR 1.09; 95% CI 0.83 to 1.42). Subgroup analysis evaluating the impact of different antibiotics showed non-significant differences for these outcomes with different durations of therapy. AUTHORS' CONCLUSIONS: The evidence of this review suggests that a short course (three days) of antibiotic therapy is as effective as a longer treatment (five days) for non-severe pneumonia in children under five years of age. However, there is a need for more well-designed RCTs to support our review findings. PMID- 18425931 TI - Targeted therapy for advanced renal cell carcinoma. AB - BACKGROUND: Advanced renal cell carcinoma has been resistant to drug therapy of different types and new types of drug therapy are needed. Targeted agents inhibit known molecular pathways involved in cellular proliferation and neoangiogenesis, the induction by the tumour of host microvascular networks. Angiogenesis is of special interest in the clear cell histologic subtype of renal cancer because of its vascularity and constitutively activated hypoxia-inducible path in the majority of tumours. OBJECTIVES: 1) To provide a systematic review of studies testing targeted agents.2) To identify the type and degree of clinical benefit, if any, of targeted agents over the prior standard of care, particularly any impact on overall survival. SEARCH STRATEGY: 1) Electronic search of CENTRAL, MEDLINE and EMBASE databases.2) Hand search of international cancer meeting abstract and other sources specified in the protocol. SELECTION CRITERIA: Randomized controlled studies of targeted agents in patients with advanced renal cell cancer reporting major remission rate or overall survival by allocation. Progression-free survival (PFS) was adopted as an additional outcome because PFS was a commonly chosen primary outcome, and because several pivotal studies allowed crossover from the control to the investigational arm after closure to accrual thereby making overall survival a problematic endpoint. DATA COLLECTION AND ANALYSIS: Nineteen fully eligible studies tested ten different targeted agents (Table 04). One additional study was excluded because no outcome data by allocation have been reported (Hutson 2007). For purposes of comparison, the studies were divided into three groups: Group 1 studies compared different doses of the same agents; Group 2 studies examined the impact of targeted agents in patients who had received prior cytokine or other systemic therapy; and Group 3 studies tested targeted agents in systemically naive patients, either against standard interferon-alfa or against another control therapy. Meta-analysis was not utilized because there were very few situations where the same agents had been tested in the same group in more than one study. MAIN RESULTS: In systemically untreated patients in studies using subcutaneous interferon-alfa as control therapy, the major findings were: 1) An improvement in overall survival has been demonstrated only with the use of weekly intravenous temsirolimus in patients with unselected renal cancer histology and adverse prognostic features (median survival 10.9 months versus 7.3 months for temsirolimus or interferon alfa respectively, HR 0.73, P = 0.008 log rank, Hudes 2007). However, the chance of major remission was low and not improved with temsirolimus. 2) In patients with mostly good or intermediate prognostic risk with clear cell renal cancer, oral sunitinib improves the chance of major remission, the probability of symptomatic improvement, and freedom from disease progression (Motzer 2007); in a similar setting, the addition of biweekly intravenous bevacizumab to interferon alfa also improved the chance of major remission and prolonged progression-free survival (Escudier 2007b); overall survival had not changed at the time of interim reporting of either study. In patients with clear cell renal cancers who had failed prior cytokine therapy, oral sorafenib gives a better quality of life than placebo as well as improved chance of being free of disease progression; overall survival may have improved but is hard to evaluate because of crossover of placebo-assigned patients after the study closed to accrual (Escudier 2007a). AUTHORS' CONCLUSIONS: Based on less than a decade of experience, some targeted agents with specified molecular targets have demonstrated clinically useful benefits over the previous standard of care for patients with advanced renal cancer. Much more research is required to fully establish the role of targeted agents in this condition. PMID- 18425932 TI - Resection versus no intervention or other surgical interventions for colorectal cancer liver metastases. AB - BACKGROUND: About one in four of patients with metastatic colorectal cancer have metastases isolated to the liver, of which 10% to 25% are eligible for ablation of the liver metastases, improving the five year survival rate. Treatments include hepatic resection and other modalities using cryosurgery and radiofrequency thermal ablation. Although new modalities allow safe ablation of liver metastases without the need for surgical intervention, there are still no clear guidelines on the appropriate management of patients with colorectal cancer and hepatic metastases. OBJECTIVES: The primary objectives were to compare resection of liver metastases to no intervention and other modalities of intervention (including cryosurgery and radiofrequency ablation) in terms of the benefits and harms for each intervention. SEARCH STRATEGY: Searches were conducted of the Cochrane Central Register of Controlled Trials, MEDLINE and EMBASE databases up to October 2006. In addition, references were scrutinized in identified eligible trials. SELECTION CRITERIA: Only randomized controlled trials reporting patients (regardless of age and sex) who had had curative surgery for adenocarcinoma of the colon or rectum, had been diagnosed with liver metastases and who were eligible for liver resection (i.e. with no evidence of primary or metastatic cancer elsewhere) were considered. DATA COLLECTION AND ANALYSIS: Two review authors independently extracted data using a form designed for this review. Discrepancies were resolved by consensus. MAIN RESULTS: Only one trial involving 123 people (87 male 36 female) was included. The data from this ten year prospective, randomized clinical trial suggest that hepatic cryosurgery is effective in the treatment of resectable and nonresectable liver metastases. The results show intra-operative tumor reduction (>/=90% or /= 10 microg/dL (>/= 0.48 micromol/l): RR 1.02 (95% CI 0.79, 1.30, I2 = 0); dichotomous >/= 15 microg/dL (>/=0.72 micromol/l): RR 0.60, 95% CI 0.33, 1.09, I2 = 0). Meta-analysis of the dichotomous data for the dust control subgroup found no evidence of effectiveness. The studies using soil abatement (removal and replacement) and combination intervention groups were not able to be meta-analysed due to substantial differences between studies. AUTHORS' CONCLUSIONS: Currently there is no evidence of effectiveness for household interventions for education or dust control measures in reducing blood lead levels in children as a population health measure. There is insufficient evidence for soil abatement or combination interventions. Further trials are required to establish the most effective intervention for prevention of lead exposure. Key elements should include longer term follow up and measures of compliance as well as performing trials in developing countries and differing socio-economic groups in developed countries. PMID- 18425935 TI - Pancreaticoduodenectomy (classic Whipple) versus pylorus-preserving pancreaticoduodenectomy (pp Whipple) for surgical treatment of periampullary and pancreatic carcinoma. AB - BACKGROUND: Pancreatic cancer is the fourth leading cause of cancer death for men and the fifth for women. The standard treatment for resectable tumours is either a classic Whipple operation or a pylorus-preserving pancreaticoduodenectomy but it is still unclear which of the two procedures is more favourable in terms of survival, mortality, complications, perioperative factors and quality of life. OBJECTIVES: Several publications pointed out both advantages and disadvantages of both techniques and the current basis of evidence remains unclear. The objective of this systematic review is to compare the effectiveness of each technique. SEARCH STRATEGY: A search was conducted to identify all published and unpublished randomised controlled trials. Trials were identified by searching the following electronic databases - The Cochrane Library, MEDLINE, EMBASE and Current Contents. Reference lists from trials selected by electronic searching were hand searched to identify further relevant trials. SELECTION CRITERIA: Randomised controlled trials (RCTs) comparing the classical Whipple (CW) with the pylorus preserving pancreaticoduodenectomy (PPW) were considered eligible if patients with periampullary or pancreatic carcinoma were included. DATA COLLECTION AND ANALYSIS: Two authors independently extracted data for included studies. A random effects model was used for pooling data from the different trials. Binary outcomes were compared using odds ratios, continuous outcomes were pooled using weighted mean differences and hazard ratios were used to for the meta-analysis of survival data. The methodological quality of included studies was evaluated independently by two authors according to quality standards and by using a questionnaire that covers different aspects of quality. MAIN RESULTS: 1235 abstracts were retrieved and checked for eligibility and seven RCTs were finally included. The critical appraisal revealed vast heterogeneity with respect to methodological quality and outcome parameters. The comparison of overall in hospital mortality (odds ratio 0.49; 95% CI 0.17 to 1.40; P=0.18), overall survival (hazard ratio 0.84; 95% CI 0.61 to 1.16; P=0.29) and morbidity showed no significant difference. However, operating time (weighted mean difference -68.26 min; 95% CI -105.70 to -30.83; P=0.0004) and intra-operative blood loss (weighted mean difference -0.76 ml; 95% CI -0.96 to -0.56; P<0.00001) were significantly reduced in the PPW group. AUTHORS' CONCLUSIONS: There is no evidence of relevant differences in mortality, morbidity and survival between the PPW and the CW. Given obvious clinical and methodological inter-study heterogeneity, future efforts have to be undertaken to perform high quality RCTs of complex surgical interventions on the basis of well defined outcome parameters. PMID- 18425937 TI - Fitness training for cardiorespiratory conditioning after traumatic brain injury. AB - BACKGROUND: Cardiorespiratory deconditioning is a common sequelae after traumatic brain injury (TBI). Clinically, fitness training is implemented to address this impairment, however this intervention has not been subject to rigorous review. OBJECTIVES: The primary objective was to evaluate whether fitness training improves cardiorespiratory fitness in people who have sustained a TBI. SEARCH STRATEGY: We searched ten electronic databases (Cochrane Injuries Group Trials Register; Cochrane Central Register of Controlled Trials (CENTRAL); EMBASE; PubMed (MEDLINE); CINAHL; AMED; SPORTDiscus; PsycINFO; PEDro and PsycBITE) and two clinical trials registers (TrialsCentral and Current Controlled Trials). The last search was August 2007. In addition we screened reference lists from included studies and contacted trialists to identify further studies. SELECTION CRITERIA: Randomised controlled studies with TBI participants were eligible if they compared an exercise programme incorporating cardiorespiratory fitness training to usual care, a non-exercise intervention or no intervention. DATA COLLECTION AND ANALYSIS: Two authors independently screened the search output, extracted data and assessed quality. All trialists were contacted for additional information. Mean difference and 95% confidence intervals (CI) were calculated for continuous data and risk difference or odds ratio and 95% CI were calculated for dichotomous data. Data were pooled when there were sufficient studies with clinical and statistical homogeneity. MAIN RESULTS: Six studies, incorporating 303 participants, were included. The participants were primarily males, in their mid thirties who had sustained a severe TBI. The studies were clinically diverse with regard to the interventions, time post-injury and the outcome measures used; therefore, the primary outcome could not be pooled. Three of the six studies indirectly assessed change in cardiorespiratory fitness after fitness training using the peak power output obtained during cycle ergometry (either at volitional fatigue or at a predetermined endpoint, that is, a percentage of predicted heart rate maximum). Cardiorespiratory fitness was improved after fitness training in one study (mean difference 59 watts, 95% CI 24 to 94), whilst there was no significant improvement in the other two studies. Four of the six studies had no drop-outs from their intervention group and no adverse events were reported in any study. AUTHORS' CONCLUSIONS: There is insufficient evidence to draw any definitive conclusions about the effects of fitness training on cardiorespiratory fitness. Whilst it appears to be a safe and accepted intervention for people with TBI, more adequately powered and well-designed studies are required to determine the effects across a range of outcome measures. PMID- 18425936 TI - Interventions for treating lymphocytic colitis. AB - BACKGROUND: Lymphocytic colitis is a cause of chronic diarrhea. Therapy is based mainly on case series and uncontrolled trials, or by extrapolation of data for treating collagenous colitis, a related disorder. This review was performed to identify therapies for lymphocytic colitis that have been proven in randomized controlled trials. OBJECTIVES: To determine effective treatments for patients with clinically active lymphocytic colitis. SEARCH STRATEGY: The MEDLINE, PUBMED and EMBASE databases were searched using the search criteria "microscopic colitis" or "lymphocytic colitis" and "treatment" or "therapy" or "management" to identify relevant papers published between 1970 and December 2007. Manual searches from the references of identified papers and relevant review papers were performed. Abstracts from major gastroenterological meetings were searched to identify research submitted in abstract form only. The trial registry website www.ClinicalTrials.gov was searched to identify registered but unpublished trials. Finally, the Cochrane Central Register of Controlled Trials and the Cochrane Inflammatory Bowel Disease and Functional Bowel Disorders Group Specialized Trials Register were searched for other studies. SELECTION CRITERIA: Five randomized controlled trials were identified. Three of these studies, which assessed bismuth subsalicylate vs. placebo, budesonide vs. placebo, and mesalazine vs. mesalazine vs. cholestyramine in treating active disease, are included in this review. DATA COLLECTION AND ANALYSIS: Data were extracted independently by each author onto 2x2 tables (treatment versus placebo or active comparator and response versus no response). For therapies assessed in one trial only, P values were derived using the chi-square test. MAIN RESULTS: Forty-one patients were enrolled in the trial studying budesonide (9 mg/day for 6 weeks versus placebo). Budesonide was more effective than placebo at inducing both clinical (P = 0.004; NNT = 3) and histological responses (P = 0.04; NNT = 3). Forty-one patients were enrolled in the study assessing mesalazine versus mesalazine plus cholestyramine. A high proportion of patients in each group responded to treatment. However, no statistically significant difference in clinical response was found between the two treatment groups (P = 0.95). Five patients were enrolled in the trial studying bismuth subsalicylate (nine 262 mg tablets daily for 8 weeks vs. placebo). There were no differences in clinical (P=0.10) or histological responses (P=0.71) in patients treated with bismuth subsalicylate compared with placebo. AUTHORS' CONCLUSIONS: A single trial studying budesonide suggests that it may be effective for the treatment of active lymphocytic colitis. An ongoing placebo-controlled trial may confirm the benefit of budesonide. There is weaker evidence that mesalazine with or without cholestyramine may be effective for the treatment of lymphocytic colitis, but this benefit needs to be confirmed in a placebo-controlled study. No conclusions can be made regarding bismuth subsalicylate. These agents require further study before they can be recommended as treatment options for lymphocytic colitis. Further trials studying interventions for lymphocytic colitis are warranted. PMID- 18425938 TI - Sustained-release naltrexone for opioid dependence. AB - BACKGROUND: Naltrexone is an opioid antagonist which effectively blocks heroin effects. Since opioid dependence treatment with naltrexone tablets suffers from high dropout rates, several depot injections and implants are under investigation. Sustained-release formulations are claimed to be effective, but a systematic review of the literature is lacking. OBJECTIVES: To evaluate the effectiveness of sustained-release naltrexone for opioid dependence and its adverse effects in different study populations. SEARCH STRATEGY: The following databases were searched from their inception to November 2007: Cochrane Central Register of Controlled Trials, MEDLINE, EMBASE, CINAHL, LILACS, PsycINFO, ISI Web of Science, trial database at http://clinicaltrials.gov, available NIDA monographs, CPDD and AAAP conference proceedings. The reference lists of identified studies, published reviews and relevant web sides were searched manually. Study authors and drug companies were contacted to obtain any unpublished material or missing data. SELECTION CRITERIA: To evaluate effectiveness only RCTs were included. To evaluate safety, any clinical trial reporting adverse effects was assessed. Treatment condition was extended to include alcohol dependent subjects and healthy volunteers. DATA COLLECTION AND ANALYSIS: Reviewers independently evaluated the reports, rated methodological quality and extracted data. Analyses were performed separately for opioid dependent, alcohol dependent and healthy participants. MAIN RESULTS: Foe effectiveness, one report met inclusion criteria. Two dosages of naltrexone depot injections (192 and 384 mg) were compared to placebo. High-dose significantly increased days in treatment compared to placebo (WMD 21.00, 95% CI 10.68 to 31.32, p<0.0001). High-dose compared to low-dose significantly increased days in treatment (WMD 12.00, 95% CI 1.69 to 22.31, p=0.02). Number of patients retained in treatment did not show significant differences between groups. For adverse effects, seventeen reports met inclusion criteria analyses, six were RCTs. Side effects were significantly more frequent in naltrexone depot groups compared to placebo. In alcohol dependent samples only, adverse effects appeared to be significantly more frequent in the low-dose naltrexone depot groups compared to placebo (RR 1.18, 95% CI 1.02 to 1.36, p=0.02). In the opioid dependent sample, group differences were not statistically significant. Reports on systematic assessment of side effects and adverse events were scarce. AUTHORS' CONCLUSIONS: There is insufficient evidence to evaluate the effectiveness of sustained-release naltrexone for treatment of opioid dependence. For naltrexone injections, administration site-related adverse effects appear to be frequent, but of moderate intensity and time limited. For a harm-benefit evaluation of naltrexone implants, more data on side effects and adverse events are needed. PMID- 18425939 TI - Exercise for the management of cancer-related fatigue in adults. AB - BACKGROUND: Cancer-related fatigue is now recognised as an important symptom associated with cancer and its treatment. A number of studies have investigated the effects of physical activity in reducing cancer-related fatigue with no definitive conclusions regarding its effectiveness. OBJECTIVES: To evaluate the effect of exercise on cancer-related fatigue both during and after cancer treatment. SEARCH STRATEGY: The Cochrane Controlled Trials Register (CENTRAL/CCTR), MEDLINE (1966 to July 2007), EMBASE (1980 to July 2007), CINAHL (1982 to July 2007), British Nursing Index (January 1984 to July 2007), AMED (1985 to July 2007), SIGLE (1980 to July 2007), and Dissertation Abstracts International (1861 to July 2007) were all searched using key words. Reference lists off all studies identified for inclusion and relevant reviews were also searched. In addition, relevant journals were hand searched and experts in the field of cancer-related fatigue were contacted. SELECTION CRITERIA: Randomised controlled trials (RCTs) that investigated the effect of exercise on cancer related fatigue in adults were included. DATA COLLECTION AND ANALYSIS: Two review authors independently assessed the methodological quality of studies and extracted data based upon predefined criteria. Where data were available meta analyses were performed for fatigue using a random-effects model. MAIN RESULTS: Twenty-eight studies were identified for inclusion (n = 2083 participants), with the majority carried out on participants with breast cancer (n = 16 studies; n = 1172 participants). A meta-analysis of all fatigue data, incorporating 22 comparisons provided data for 920 participants who received an exercise intervention and 742 control participants. At the end of the intervention period exercise was statistically more effective than the control intervention (SMD 0.23, 95% Confidence Interval (CIs) -0.33 to -0.13). AUTHORS' CONCLUSIONS: Exercise can be regarded as beneficial for individuals with cancer-related fatigue during and post cancer therapy. Further research is required to determine the optimal type, intensity and timing of an exercise intervention. PMID- 18425940 TI - Antenatal lower genital tract infection screening and treatment programs for preventing preterm delivery. AB - BACKGROUND: Preterm birth is birth before 37 weeks' gestation. Genital tract infection is one of the causes of preterm birth. Infection screening during pregnancy has been used to reduce preterm birth. However, infection screening may have some adverse effects, e.g. increased antibiotic drug resistance, increased costs of treatment. OBJECTIVES: To assess the effectiveness and complications of antenatal lower genital tract infection screening and treatment programs in reducing preterm birth and subsequent morbidity. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group's Trials Register (January 2008) and the Cochrane Central Register of Controlled Trials (The Cochrane Library 2007, Issue 2). SELECTION CRITERIA: We included all published and unpublished randomised controlled trials in any language that evaluated any described methods of antenatal lower genital tract infection screening compared with no screening. Preterm births have been reported as an outcome. DATA COLLECTION AND ANALYSIS: Two review authors independently assessed eligibility, trial quality and extracted data. MAIN RESULTS: One study (4155 women) met the inclusion criteria. This trial is of high methodological quality. In the intervention group (2058 women), the results of infection screening and treatment for bacterial vaginosis, trichomonas vaginalis and candidiasis were reported; in the control group (2097 women), the results of the screening program for the women allocated to receive routine antenatal care were not reported. Preterm birth before 37 weeks was significantly lower in the intervention group (3% versus 5% in the control group) with a relative risk (RR) of 0.55 (95% confidence interval (CI) 0.41 to 0.75). The incidence of preterm birth for low birthweight preterm infants with a weight equal to or below 2500 g and very low birthweight infants with a weight equal to or below 1500 g were significantly lower in the intervention group than in the control group (RR 0.48, 95% CI 0.34 to 0.66 and RR 0.34; 95% CI 0.15 to 0.75, respectively). AUTHORS' CONCLUSIONS: There is evidence that infection screening and treatment programs in pregnant women may reduce preterm birth and preterm low birthweights. Future trials should evaluate the effects of types of infection screening program, gestational ages at screening test and the costs of introducing an infection screening program. PMID- 18425941 TI - Ciclesonide versus placebo for chronic asthma in adults and children. AB - BACKGROUND: Inhaled corticosteroids are an integral part of asthma management, and act as an anti-inflammatory agent in the airways of the lung. These agents confer significant benefit in terms of symptom management and improvement in lung function, but may also cause harm in terms of local and systemic side-effects. Ciclesonide is a novel steroid that has efficient distribution and release properties that mean it can be taken once daily, making it potentially useful in ongoing asthma management. OBJECTIVES: To assess the efficacy of inhaled ciclesonide in adults and children with chronic asthma. SEARCH STRATEGY: We searched the Cochrane Airways Group register of trials with pre-defined terms. Additional searches of CENTRAL and PubMed were undertaken. The literature searches for this review are current up to June 2007. SELECTION CRITERIA: Randomised parallel or crossover studies were eligible for the review. We included studies comparing ciclesonide with placebo, and we also included studies comparing ciclesonide at different doses. DATA COLLECTION AND ANALYSIS: Two authors assessed studies for inclusion in the review, extracted data independently and checked each others' work. We contacted study investigators in order to obtain additional data. Extracted data were entered into RevMan 4.2 and analysed as fixed effect mean differences for continuous data, and fixed effect risk ratios for dichotomous data. MAIN RESULTS: Eighteen trials (reporting 20 study comparisons) met the review entry criteria. We report findings from 18 group comparisons where data were available (6343 participants, of whom 1692 were children). Ciclesonide versus placebo: The short duration of the included studies means that there is a lack of data with respect to the impact of ciclesonide on asthma exacerbations. At doses of 100 mcg/d or less up to 400 mcg/d in mild to moderate asthma, ciclesonide improved lung function, asthma symptoms and rescue inhaler use, compared with placebo.Dose response outcomes: Comparisons of 100 versus 200 mcg/d, 100 versus 400 mcg/d and 400 versus 800 mcg/d did not yield significant differences in lung function outcomes. Adverse event data were not available in sufficient detail to permit assessment of the safety profile of this drug. AUTHORS' CONCLUSIONS: Ciclesonide was more effective than placebo, in the short term, in improving lung function in patients with mild to moderate asthma previously treated with inhaled corticosteroids. There remain questions as to dose response, and the lack of data on the longer term impact on exacerbations and safety profile should be addressed in future studies. PMID- 18425942 TI - Interventions to improve occupational health in depressed people. AB - BACKGROUND: Work disability such as sickness absence is common in people with depression. OBJECTIVES: To evaluate the effectiveness of interventions aimed at reducing work disability in depressed workers. SEARCH STRATEGY: We searched the CCDANCTR-Studies and CCDANCTR-References on 2/8/2006, Cochrane Library CENTRAL register, MEDLINE, EMBASE, CINAHL, PsycINFO, OSH-ROM (Occupational Safety and Health), NHS-EED, and DARE. SELECTION CRITERIA: We included randomised controlled trials (RCTs) and cluster RCTs of work-directed and worker-directed interventions for depressed people, using sickness absence as the primary outcome DATA COLLECTION AND ANALYSIS: Two authors independently extracted data and assessed trial quality. We used standardised mean differences (SMD) with 95% confidence intervals (CIs) to pool study results where possible. MAIN RESULTS: We included eleven studies, all of worker-directed interventions, involving 2556 participants. Only one study addressed work issues using adjuvant occupational therapy. Other interventions evaluated anti-depressant medication (selective serotonin reuptake inhibitors, serotonin-norepinephrine reuptake inhibitors, tricyclic antidepressants, monoamino-oxidase inhibitors), psychodynamic therapy, enhanced primary care and psychological treatment. For medication, the combined results of three studies (n=864) showed no difference between antidepressant medication and alternative medication in their effect on days of sickness absence (SMD 0.09; 95% CI -0.05 to 0.23) In two pooled studies (n=969), the effect of enhanced primary care on days of sickness absence did not differ from usual care in the medium term (SMD -0.02; 95% CI -0.15 to 0.12). All other comparisons were based on single studies (n=6), all of which showed a lack of significant difference for sickness absence between groups, with the exception of one small study, combined psychodynamic therapy and TCAs versus TCAs alone, which favoured the combined treatment. AUTHORS' CONCLUSIONS: Based on a heterogeneous sample of studies, there is currently no evidence of an effect of medication alone, enhanced primary care, psychological interventions or the combination of those with medication on sickness absence of depressed workers. In future RCTs, interventions should specifically address work issues, and occupational outcomes should be used to measure the effect.. PMID- 18425943 TI - Immediate start of hormonal contraceptives for contraception. AB - BACKGROUND: Health care providers often tell women to wait until the next menses to begin hormonal contraception. The main intent is to avoid contraceptive use during an undetected pregnancy. An alternative is to start hormonal contraception immediately with back-up birth control for the first seven days. Immediate initiation was first introduced with combined oral contraceptives (COCs), and has expanded to other hormonal contraceptives. How immediate start compares to conventional menses-dependent start is unclear regarding effectiveness, continuation, and acceptability. The immediate-start approach may improve women's access to, and continuation of, hormonal contraception. OBJECTIVES: This review examined randomized controlled trials of immediate-start hormonal contraception for differences in effectiveness, continuation, and acceptability. SEARCH STRATEGY: We searched MEDLINE, CENTRAL, POPLINE, EMBASE, and LILACS for trials of immediate-start hormonal contraceptives. We contacted researchers to find other studies. SELECTION CRITERIA: We included randomized controlled trials that compared immediate start to conventional start of hormonal contraception. Also included were trials that compared immediate start of different hormonal contraceptive methods with each other. DATA COLLECTION AND ANALYSIS: Data were abstracted by two authors and entered into RevMan. The Peto odds ratio (OR) with 95% confidence interval (CI) was calculated. MAIN RESULTS: Five studies were included. Method discontinuation was similar between groups in all trials. Bleeding patterns and side effects were similar in trials that compared immediate with conventional start. In a study of depot medroxyprogesterone acetate (DMPA), immediate start of DMPA showed fewer pregnancies than a 'bridge' method before DMPA (OR 0.36; 95% CI 0.16 to 0.84). Further, more women in the immediate-DMPA group were very satisfied versus those with a 'bridge' method (OR 1.99; 95% CI 1.05 to 3.77).A trial of two immediate-start methods showed the vaginal ring group had less prolonged bleeding (OR 0.42; 95% CI 0.20 to 0.89) and less frequent bleeding (OR 0.23; 95% CI 0.05 to 1.03) than COC users. The ring group also reported fewer side effects. For satisfaction, more immediate ring users were very satisfied than immediate COC users (OR 2.88; 95% CI 1.59 to 5.22). AUTHORS' CONCLUSIONS: We found limited evidence that immediate start of hormonal contraception reduces unintended pregnancies or increases method continuation. However, the pregnancy rate was lower with immediate start of DMPA versus another method. Some differences were associated with contraceptive type rather than initiation method, that is, immediate ring versus immediate COC. More studies are needed of immediate versus conventional start of the same hormonal contraceptive. PMID- 18425944 TI - Medically assisted hydration for palliative care patients. AB - BACKGROUND: Many palliative care patients have reduced oral intake during their illness. The management of this can include the provision of medically assisted hydration with the aim of prolonging the length of life of a patient, improving their quality of life, or both. OBJECTIVES: To determine the effect of medically assisted hydration in palliative care patients on their quality and length of life. SEARCH STRATEGY: Studies were identified from searching CENTRAL, MEDLINE (1966 to 2008), EMBASE (1980 to 2008), CINAHL, CANCERLIT, Caresearch, Dissertation abstracts, SCIENCE CITATION INDEX and the reference lists of all eligible studies, key textbooks, and previous systematic reviews. The date of the latest search was February 2008. SELECTION CRITERIA: All relevant randomised controlled trials (RCTs) or prospective controlled studies of medically assisted hydration in palliative care patients. DATA COLLECTION AND ANALYSIS: Five relevant studies were identified. These included two RCTs (93 participants), and three prospective controlled trials (360 participants). These were assessed independently by two review authors for quality and validity. The small number of studies and the heterogeneity of the data meant that a quantitative analysis was not possible, so a description of the main findings was included only. MAIN RESULTS: One study found that sedation and myoclonus (involuntary contractions of muscles) were improved more in the intervention group (28 - hydration, 23 - placebo). Another study found that dehydration was significantly higher in the non-hydration group, but that some fluid retention symptoms (pleural effusion, peripheral oedema and ascites) were significantly higher in the hydration group (59 - hydration group, 167 - non -hydration group). The other three studies did not show significant differences in outcomes between the two groups. AUTHORS' CONCLUSIONS: There are insufficient good quality studies to make any recommendations for practice with regard to the use of medically assisted hydration in palliative care patients. PMID- 18425945 TI - Intravenous versus inhalation anaesthesia for one-lung ventilation. AB - BACKGROUND: The technique called one-lung ventilation can confine bleeding or infection to one lung, prevent rupture of a lung cyst or, more commonly, facilitate surgical exposure of the unventilated lung. During one-lung ventilation, anaesthesia is maintained either by delivering a volatile anaesthetic to the ventilated lung or by infusing an intravenous anaesthetic. It is possible that the method chosen to maintain anaesthesia may affect patient outcomes. OBJECTIVES: The objective of this review was to evaluate the effectiveness and safety of intravenous versus inhalation anaesthesia for one lung ventilation. SEARCH STRATEGY: We searched the Cochrane Central Register of Controlled Trials (CENTRAL) (The Cochrane Library 2006, Issue 3), MEDLINE, LILACS, EMBASE (from inception to June 2006), ISI web of Science (1945 to June 2006), reference lists of identified trials, and bibliographies of published reviews. We also contacted researchers in the field. There were no language restrictions. SELECTION CRITERIA: We included randomized controlled trials and quasi-randomized controlled trials of intravenous versus inhalation anaesthesia for one-lung ventilation. DATA COLLECTION AND ANALYSIS: Two authors independently assessed trial quality and extracted data. We contacted study authors for additional information. MAIN RESULTS: We included nine studies that enrolled 291 participants. We could not perform meta-analyses as the included studies did not report the outcomes listed in the protocol for this review. AUTHORS' CONCLUSIONS: There is no evidence from randomized controlled trials of differences in patient outcomes for anaesthesia maintained by intravenous versus inhalational anaesthesia during one-lung ventilation. This review highlights the need for continued research into the use of intravenous versus inhalation anaesthesia for one-lung ventilation. Future trials should have standardized outcome measures such as death, adverse postoperative outcomes and intraoperative awareness. Dropouts and losses to follow up should be reported. PMID- 18425946 TI - Maintenance agonist treatments for opiate dependent pregnant women. AB - BACKGROUND: The prevalence of opiate use among pregnant women ranges from 1% to 2% to as much as 21%. Heroin crosses the placenta and pregnant opiate dependent women experience a six fold increase in maternal obstetric complications such as low birth weight, toxaemia, 3rd trimester bleeding, malpresentation, puerperal morbidity, fetal distress and meconium aspiration. Neonatal complications include narcotic withdrawal, postnatal growth deficiency, microcephaly, neurobehavioral problems, increased neonatal mortality and a 74-fold increase in sudden infant death syndrome. OBJECTIVES: To assess the effectiveness of any maintenance treatment alone or in combination with psychosocial intervention compared to no intervention, other pharmacological intervention or psychosocial interventions on child health status, neonatal mortality, retaining pregnant women in treatment, and reducing use of substances SEARCH STRATEGY: We searched Cochrane Drugs and Alcohol Group' Register of Trials (June 2007), PubMed (1966 - June 2007), CINAHL (1982- June 2007), reference lists of relevant papers, sources of ongoing trials, conference proceedings, National focal points for drug research. Authors of included studies and experts in the field were contacted. SELECTION CRITERIA: Randomised controlled trials enrolling opiate dependent pregnant women DATA COLLECTION AND ANALYSIS: The authors assessed independently the studies for inclusion and methodological quality. Doubts were solved by discussion. MAIN RESULTS: We found three trials with 96 pregnant women. Two compared methadone with buprenorphine and one methadone with oral slow morphine. For the women there was no difference in drop out rate RR 1.00 (95% CI 0.41 to 2.44) and use of primary substance RR 2.50 (95% CI 0.11 to 54.87) between methadone and buprenorphine, whereas oral slow morphine seemed superior to methadone in abstaining women from the use of heroin RR 2.40 (95% CI 1.00 to 5.77)For the newborns in one trial buprenorphine performed better than methadone for birth weight WMD -530 gr (95% CI -662 to -397), this result is not confirmed in the other trial. For the APGAR score both studies didn't find significant difference . No differences for NAS measures used. Comparing methadone with oral slow morphine no differences for birth weight and mean duration of NAS. The APGAR score wasn't considered. AUTHORS' CONCLUSIONS: We didn't find any significant difference between the drugs compared both for mother and for child outcomes; the trials retrieved were too few and the sample size too small to make firm conclusion about the superiority of one treatment over another. There is an urgent need of big randomized controlled trials. PMID- 18425947 TI - Mu-opioid antagonists for opioid-induced bowel dysfunction. AB - BACKGROUND: Opioid-induced bowel dysfunction (OBD) is characterized by constipation, incomplete evacuation, bloating, and increased gastric reflux. OBD occurs both acutely and chronically, in multiple disease states, resulting in increased morbidity and reduced quality of life. OBJECTIVES: To compare the efficacy and safety of traditional and peripherally active opioid antagonists versus conventional interventions for OBD. SEARCH STRATEGY: We searched MEDLINE, the Cochrane Central Register of Controlled Trials and EMBASE in January 2007. Additional reports were identified from the reference lists of retrieved papers. SELECTION CRITERIA: Studies were included if they were randomized controlled trials that investigated the efficacy of mu-opioid antagonists for OBD. DATA COLLECTION AND ANALYSIS: Data were extracted by two independent review authors and included demographic variables, diagnoses, interventions, efficacy, and adverse events. MAIN RESULTS: Twenty-three studies met inclusion criteria and provided data on 2871 opioid antagonist-treated patients. The opioid antagonists investigated were alvimopan (nine studies), methylnaltrexone (six), naloxone (seven), and nalbuphine (one). Meta-analysis demonstrated that methylnaltrexone and alvimopan were better than placebo in reversing opioid-induced increased gastrointestinal transit time and constipation, and that alvimopan appears to be safe and efficacious in treating postoperative ileus. The incidence of adverse events with opioid antagonists was similar to placebo and generally reported as mild-to-moderate. AUTHORS' CONCLUSIONS: Insufficient evidence exists for the safety or efficacy of naloxone or nalbuphine in the treatment of OBD. Long-term efficacy and safety of any of the opioid antagonists is unknown, as is the incidence or nature of rare adverse events. Alvimopan and methylnaltrexone both show promise in treating OBD, but further data will be required to fully assess their place in therapy. PMID- 18425948 TI - Chinese herbal medicine Huangqi type formulations for nephrotic syndrome. AB - BACKGROUND: At present, there is a lack of safe and effective drugs for nephrotic syndrome (NS). Huangqi type formulations have been used to treat nephrotic syndrome for years in China, however the effects and safety of these formulations have not been systematically reviewed. OBJECTIVES: To assess the benefits and harms of Huangqi and Huangqi type formulations in treating NS in any age group, either as sole agents or in addition to other drug therapies. SEARCH STRATEGY: We searched the Cochrane Central Register of Controlled Trials (CENTRAL), MEDLINE, EMBASE, Chinese Biomedicine Database (CBM), CNKI, VIP and reference lists of articles. There was no language restriction. Date of most recent search: June 2006. SELECTION CRITERIA: All randomised controlled trials (RCTs) assessing the use of Huangqi or Huangqi type formulations in treating NS in adults and children, either as sole agents or in addition to other drug therapies. DATA COLLECTION AND ANALYSIS: Two authors independently assessed study quality and extracted data. For dichotomous outcomes (remission, side effects and Inefficacy rate), results were expressed as relative risk (RR) and 95% confidence intervals (CI). Continuous outcomes (triglycerides cholesterol, plasma albumin) results were expressed as mean difference (WMD) with 95% CI. MAIN RESULTS: Three studies were identified (n = 128), all comparing Huangqi type formulations with placebo. Huangqi injection had a positive effect on plasma albumin (WMD 6.90, 95% Cl 3.60 to 10.20) and cholesterol (WMD 2.13, 95% Cl -2.97 to -1.29). Huangqi and red Chinese date reduced some adverse reactions (Cushing's syndrome: RR 0.55, 95% Cl 0.32 to 0.94; hormone reduced syndrome: RR 0.58, 95% Cl 0.39 to 0.85, respiratory tract infection: RR 0.27, 95% Cl 0.08 to 0.88), but no benefit on reducing relapse. Huangqi and Danggui had a positive effect on cholesterol (WMD -0.85, 95% Cl -1.70 to 0.00). AUTHORS' CONCLUSIONS: Huangqi type formulations may have some positive effects in treating NS by increasing plasma albumin and reducing blood cholesterol, Cushing's syndrome, hormone reduced syndrome and respiratory tract infection. However, limited by the lack of high quality clinical studies, we are unable to recommend Huangqi type formulations for NS. Large, properly randomised, placebo-controlled, double-blind studies are required. PMID- 18425949 TI - Tailored interventions based on exhaled nitric oxide versus clinical symptoms for asthma in children and adults. AB - BACKGROUND: The measurement of severity and control of asthma in both children and adults can be based on subjective or objective measures. It has been advocated that fractional exhaled nitric oxide (FeNO) can be used to monitor airway inflammation as it correlates with some markers of asthma. Interventions for asthma therapies have been traditionally based on symptoms and/or spirometry. OBJECTIVES: To evaluate the efficacy of tailoring asthma interventions based on exhaled nitric oxide in comparison to clinical symptoms (with or without spirometry/peak flow) for asthma related outcomes in children and adults. SEARCH STRATEGY: We searched the Cochrane Airways Group Specialised Register of Trials, the Cochrane Central Register of Controlled Trials (CENTRAL), MEDLINE, EMBASE and reference lists of articles. The last search was completed in December 2006. SELECTION CRITERIA: All randomised controlled comparisons of adjustment of asthma therapy based on exhaled nitric oxide compared to traditional methods (primarily clinical symptoms and spirometry/peak flow). DATA COLLECTION AND ANALYSIS: Results of searches were reviewed against pre-determined criteria for inclusion. Relevant studies were independently selected in duplicate. Two authors independently assessed trial quality and extracted data. Authors were contacted for further information but none were received. Data was analysed as "intervention received" and sensitivity analyses performed. MAIN RESULTS: Four (2 adult and 2 paediatric) studies were included; these studies differed in a variety of ways including definition of asthma exacerbations, FeNO cut off levels and duration of study. Of 356 participants randomised, 324 completed the trials. In the meta-analysis, there was no difference between groups for the primary outcome of asthma exacerbations or for other outcomes (clinical symptoms, FeNO level and spirometry). In post-hoc analysis, a significant reduction in mean final daily dose inhaled corticosteroid per adult was found in the group where treatment was based on FeNO in comparison to clinical symptoms; WMD -282.46 (95% CI -422.08 to -142.84). There was no difference in ICS dose between the groups in the overall daily dose in the adult studies or in the paediatric studies. AUTHORS' CONCLUSIONS: Tailoring the dose of inhaled corticosteroids based on exhaled nitric oxide in comparison to clinical symptoms was carried out in different ways in the four studies that were found, and the results show only modest differences. The role of utilising exhaled nitric oxide to tailor the dose of inhaled corticosteroids is currently uncertain. PMID- 18425950 TI - Financial benefits for child health and well-being in low income or socially disadvantaged families in developed world countries. AB - BACKGROUND: A strong and consistent relationship has been observed between relative poverty and poor child health and wellbeing even among rich nations. This review set out to examine evidence that additional monies provided to poor or disadvantaged families may benefit children by reducing relative poverty and thereby improving children's health, well-being and educational attainment. OBJECTIVES: To assess the effectiveness of direct provision of additional monies to socially or economically disadvantaged families in improving children's health, well-being and educational attainment SEARCH STRATEGY: In total 10 electronic databases were searched including the Cochrane library searched 2006 (Issue 1), Medline searched 1966 to May 2006 , Econlit searched 1969 to June 2006 and PsycINFO searched 1872 to June 2006, together with 3 libraries of working papers (MDRC, SSRN, SRDC). The general search strategy was [terms for income and financial benefits] and [paediatric terms] and [RCT filter] SELECTION CRITERIA: Studies selected provided money to relatively poor families (which included a child under the age of 18 or a pregnant woman), were randomised or quasi randomised, measured outcomes related to child health or wellbeing and were conducted in a high income country. DATA COLLECTION AND ANALYSIS: Titles and abstracts identified in the search were independently assessed for eligibility by two reviewers. Data were extracted and entered into RevMan, synthesised and presented in both written and graphical form (forest plots). MAIN RESULTS: Nine trials including more than 25,000 participants were included in this review. No effect was observed on child health, measures of child mental health or emotional state. Non-significant effects favouring the intervention group were seen for child cognitive development and educational achievement, and a non-significant effect favouring controls in rates of teenage pregnancy. AUTHORS' CONCLUSIONS: The review set out to examine the potential of financial support to poor families to improve circumstances for children. However, on the basis of current evidence we can not state unequivocally whether financial benefits delivered as an intervention are effective at improving child health or wellbeing in the short term. Our conclusions are limited by the fact that most of the studies had small effects on total household income and that while no conditions were attached to how money was spent, all studies included strict conditions for receipt of payments. We note particular concerns by some authors that sanctions and conditions (such as working hours) placed on families may increase family stress. PMID- 18425951 TI - Paliperidone for schizophrenia. AB - BACKGROUND: Paliperidone, risperidone's active metabolite, is now available in an oral formulation for daily use, and an intramuscular formulation for monthly administration may follow shortly. OBJECTIVES: To compare effects of oral paliperidone with any other treatment for people with schizophrenia and schizophrenia-like illnesses. SEARCH STRATEGY: We searched the Cochrane Schizophrenia Group's Register (December 2006), and inspected references of identified studies for further trials. We contacted the manufacturers of paliperidone, the Food and Drug Administration, and authors of relevant trials for additional material. SELECTION CRITERIA: We included all relevant randomised trials. DATA COLLECTION AND ANALYSIS: We independently selected and critically appraised studies, extracted data and analysed on an intention-to-treat basis. Where possible and appropriate, we calculated risk ratios (RR) and their 95% confidence intervals (CI) with the number needed to treat (NNT). We calculated Weighted Mean Differences (WMD) for continuous data. MAIN RESULTS: Five studies compared paliperidone with placebo. Fewer people left the studies early if they were randomized to paliperidone (n=1647, 5 RCTs, RR 0.68 CI 0.61 to 0.76, NNT 7 CI 6 to 9) and those receiving any dose of paliperidone were significantly more likely to have an improvement in global state (n=1420, 4RCTs, RR 0.69 CI 0.63 to 0.75, NNT 5 CI 4 to 6). People randomised to paliperidone were less likely to experience a recurrence of psychosis (n=1638, 5 RCTs, RR 0.45 CI 0.31 to 0.66, NNT 16 CI 13 to 26) than those allocated to placebo. Adverse effect data were not well reported but paliperidone does seem to produce a greater incidence of tachycardia than placebo (n=1638, 5 RCTs, RR1.88 CI 1.28 to 2.76, NNH 21 CI 11 to 90) and a consistent, significant elevation in serum prolactin was found for both men (n=413, 3 RCTs, WMD 27.68 CI 23.66 to 31.69) and women (n=252, 3 RCTs, WMD 87.39 CI 74.27 to 100.51). People receiving paliperidone were more likely to experience extrapyramidal disorders (n=1638, 5 RCTs, RR 2.21 CI 1.26 to 3.88, NNH 28 CI 12 to 129) and weight gain (n=769, 4 RCTs, WMD 1.07 CI 0.65 to 1.49, I squared 78%) compared with those allocated to placebo. When compared with 10 mg/day olanzapine we found no differences between paliperidone and olanzapine for leaving in the short term (n=1332, 3 RCTs, RR 1.04 CI 0.89 to 1.21; 40% in both groups left by six weeks). Those receiving any dose of paliperidone were no more likely to have a recurrence of psychotic symptoms than those receiving 10 mg/day olanzapine (n=1327, 3 RCTs, RR 0.1.07 CI 0.64 to 1.76). Data from all three studies found paliperidone was less likely to produce a weight change than olanzapine (n=660, 3 RCTs, WMD -0.88 CI -1.38 to -0.37). Results for various movement disorders all favoured olanzapine. There are no clear data relating to social functioning, services use, quality of life, satisfaction and cost. AUTHORS' CONCLUSIONS: In short-term studies, oral paliperidone is an antipsychotic that is more efficacious than placebo. We found its adverse effects to be similar to those of its parent compound, risperidone, with movement disorders, weight gain, and tachycardia all more common with paliperidone than placebo. In addition, paliperidone is associated with substantial increases in serum prolactin that may be associated with sexual dysfunction, although sexual functioning outcomes were not reported. At doses greater than 3 mg per day, oral paliperidone appears comparable in efficacy to oral olanzapine 10 mg per day. Regarding the critical comparison of oral paliperidone to risperidone, we have no information and are thus unable to determine if paliperidone has any advantages or disadvantages compared to its well-known parent compound. PMID- 18425952 TI - Interventions for strabismic amblyopia. AB - BACKGROUND: Amblyopia is reduced visual acuity in one or both eyes in the absence of any demonstrable abnormality of the visual pathway. It is not immediately resolved by the correction of refractive error. Strabismus develops in approximately 5% to 8% of the general population. The aim of treatment for amblyopia is to obtain the best possible level of vision in the amblyopic eye. Different treatment options were examined within the review. OBJECTIVES: By reviewing available evidence we wanted to establish the most effective treatment for strabismic amblyopia. In particular this review aimed to examine the impact of conventional occlusion therapy for strabismic amblyopia and analyse the role of partial occlusion and optical penalisation for strabismic amblyopia. SEARCH STRATEGY: We searched CENTRAL, MEDLINE, EMBASE and LILACS in October 2007. There were no language or date restrictions in the electronic searches for trials. SELECTION CRITERIA: We included randomised controlled trials (RCTs) for the treatment of strabismic amblyopia including participants of any age. DATA COLLECTION AND ANALYSIS: Two authors working independently extracted and entered data into Review Manager 4.2 using the double data entry facility to check for errors. MAIN RESULTS: We included two RCTs. The studies reported mean logMAR visual acuity achieved. Mean difference in visual acuity was calculated. When comparing conventional part-time occlusion (with any necessary glasses), PEDIG 2006 reported this treatment was more beneficial than glasses alone for strabismic amblyopia. Mean difference between groups was -0.18 (statistically significant 95% CI -0.32 to -0.04). Supplementing occlusion therapy with near activities may produce a better visual outcome compared to non-near activities after four weeks of treatment (PEDIG 2005). Mean difference between groups was 0.17 (95% CI -0.53 to 0.19). However, this data is from a pilot study and the full data set will be added to updates of the review when available. AUTHORS' CONCLUSIONS: Occlusion, whilst wearing necessary refractive correction, appears to be more effective than refractive correction alone in the treatment of strabismic amblyopia. Combining occlusion and refractive correction with near activities may be more effective than occlusion and refractive correction alone. Further study of the role of near activities is necessary before a more definitive conclusion can be made. Results of a full trial are expected within the coming year. No RCTs were found that assessed the role of either partial occlusion or optical penalisation to refractive correction for strabismic amblyopia. PMID- 18425953 TI - Antiplatelet drugs for polycythaemia vera and essential thrombocythaemia. AB - BACKGROUND: Polycythaemia vera and essential thrombocythaemia are chronic Philadelphia-negative myeloproliferative disorders, which increase the risk of arterial and venous thrombosis as well as bleeding. In addition to the different therapeutic strategies available, aspirin is often used to prevent platelet aggregation. OBJECTIVES: To quantify the benefit and harm of antiplatelet drugs for long-term primary and secondary prophylaxis of arterial and venous thrombotic events in patients with polycythaemia vera or essential thrombocythaemia. SEARCH STRATEGY: Our searched included the CENTRAL (The Cochrane Library, issue 1 2007), MEDLINE (1966 to 2007) and EMBASE (1980 to 2007) databases, online registers of ongoing trials and conference proceedings. The date of the last search was March 2007. SELECTION CRITERIA: All randomised controlled trials (RCTs) comparing long term (>6 months) use of an antiplatelet drug versus placebo or no treatment in patients with polycythaemia vera or essential thrombocythaemia, diagnosed by established international criteria, with data for at least one of the selected outcomes, were included. DATA COLLECTION AND ANALYSIS: Using a predefined extraction form, we collected and analysed the following data where appropriate: mortality from arterial and venous thrombotic events, mortality from bleeding episodes, fatal and non-fatal arterial thrombotic events, fatal and non-fatal venous thrombotic events, micro-circulation events, transient neurological and ocular manifestations, major and minor bleeding episodes, all-cause mortality and any adverse events. We based quantitative analysis of outcome data on an intention-to-treat principle. The overall treatment effect was estimated by the pooled odds ratio (OR) with 95% confidence interval (CI) using a fixed-effect model (Mantel-Haenszel). MAIN RESULTS: Two RCTs that investigated 630 patients with an established diagnosis of polycythaemia vera, with no clear indication or contraindication to aspirin therapy, were included in this review. The use of aspirin, compared with placebo, was associated with a lower risk of fatal thrombotic events (although this benefit was not statistically significant (OR 0.20, 95% CI 0.03 to 1.14)) and did not increase the risk of major bleeding (OR 0.99, 95% CI 0.23 to 4.36). No studies have been published in patients with essential thrombocythaemia or studying other antiplatelet drugs. AUTHORS' CONCLUSIONS: The available evidence suggests that the use of aspirin is associated with a statistically non-significant reduction in the risk of fatal thrombotic events, without an increased risk of major bleeding, when compared with no treatment in patients polycythaemia vera who have no clear indication or contraindication to aspirin therapy. PMID- 18425954 TI - Dopamine agonist therapy in early Parkinson's disease. AB - BACKGROUND: Dopamine agonists are being used increasingly as first line treatment for Parkinson's disease, but there remains uncertainty about their clinical and cost-effectiveness relative to levodopa. OBJECTIVES: This meta-analysis aims to quantify more reliably the benefits and risks of dopamine agonists compared to placebo or levodopa in early Parkinson's disease. SEARCH STRATEGY: We searched CENTRAL (The Cochrane Library), MEDLINE, EMBASE, PubMed, LILACS and Web of Science, plus major journals in the field, abstract books, conference proceedings and reference lists of retrieved publications. SELECTION CRITERIA: Randomised trials comparing an orally administered dopamine agonist (with or without levodopa) versus placebo or levodopa or both placebo and levodopa in participants with early Parkinson's disease. DATA COLLECTION AND ANALYSIS: Two authors independently extracted data on clinician-rated disability, motor complications, other side-effects, treatment concordance, levodopa dose and mortality. MAIN RESULTS: Twenty-nine eligible trials, involving 5247 participants, were identified. Participants randomised to a dopamine agonist were less likely to develop dyskinesia (odds ratio (OR) 0.51, 95% confidence interval (CI) 0.43 to 0.59; P < 0.00001), dystonia (OR 0.64, 95% CI 0.51 to 0.81; P = 0.0002) and motor fluctuations (OR 0.75, 95% CI 0.63 to 0.90; P = 0.002) than levodopa-treated participants. However, various 'non-motor' side-effects, including oedema (OR 3.68, 95% CI 2.62 to 5.18; P < 0.00001), somnolence (OR 1.49, 95% CI 1.12 to 2.00; P = 0.007), constipation (OR 1.59, 95% CI 1.11 to 2.28; P = 0.01), dizziness (OR 1.45, 95% CI 1.09 to 1.92; P = 0.01), hallucinations (OR 1.69, 95% CI 1.13 to 2.52; P = 0.01) and nausea (OR 1.32, 95% CI 1.05 to 1.66; P = 0.02) were all increased in agonist-treated participants (compared with levodopa treated participants). Agonist-treated participants were also significantly more likely to discontinue treatment due to adverse events (OR 2.49, 95% CI 2.08 to 2.98; P < 0.00001). Finally symptomatic control of Parkinson's disease was better with levodopa than with agonists, but data were reported too inconsistently and incompletely to meta-analyse. AUTHORS' CONCLUSIONS: This meta-analysis confirms that motor complications are reduced with dopamine agonists compared to levodopa, but also establishes that other important side-effects are increased and symptom control is poorer with agonists. Larger, long-term comparative trials assessing patient-rated quality of life are needed to assess more reliably the balance of benefits and risks of dopamine agonists compared to levodopa. PMID- 18425955 TI - Abdominal lift for laparoscopic cholecystectomy. AB - BACKGROUND: Laparoscopic cholecystectomy is the main method of treatment of symptomatic gallstones. Several cardiopulmonary changes (decreased cardiac output, pulmonary compliance, and increased peak airway pressure) occur during pneumoperitoneum. These changes may not be tolerated in individuals with poor cardiopulmonary reserve. OBJECTIVES: To assess the benefits and harms of abdominal wall lift compared to pneumoperitoneum in patients undergoing laparoscopic cholecystectomy. SEARCH STRATEGY: We searched The Cochrane Hepato Biliary Group Controlled Trials Register, The Cochrane Central Register of Controlled Trials in The Cochrane Library, MEDLINE, EMBASE, and Science Citation IndexExpanded until January 2007. SELECTION CRITERIA: We included all randomised clinical trials comparing abdominal wall lift (with or without pneumoperitoneum) and pneumoperitoneum. DATA COLLECTION AND ANALYSIS: We calculated the relative risk (RR), weighted mean difference (WMD) or standardised mean difference (SMD) with 95% confidence intervals (CI) based on intention-to-treat analysis with both the fixed-effect and the random-effects model using RevMan Analysis. MAIN RESULTS: Abdominal wall lift with pneumoperitoneum versus pneumoperitoneum. A total of 156 participants (all with low anaesthetic risk) who underwent elective laparoscopic cholecystectomy were randomised in six trials to abdominal wall lift with pneumoperitoneum (n = 65) versus pneumoperitoneum only (n = 66). One trial which included 25 patients did not state the number of patients in each group. All six trials were of high risk of bias. The cardiopulmonary changes were less in abdominal wall lift than pneumoperitoneum. There was no difference in the morbidity and pain between the groups. Abdominal wall lift versus pneumoperitoneum. A total of 550 participants (the majority with low anaesthetic risk) who underwent elective laparoscopic cholecystectomy were randomised in fourteen trials to abdominal wall lift without pneumoperitoneum (n = 268) versus pneumoperitoneum (n = 282). Two of these fourteen trials were of low risk of bias. The cardiopulmonary changes were less in abdominal wall lift than with pneumoperitoneum. There was no difference in the morbidity and pain between the groups. The operating time was prolonged in abdominal wall lift compared with pneumoperitoneum (WMD 7.74, 95% CI 1.37 to 14.12). AUTHORS' CONCLUSIONS: (1) Abdominal wall lift seems safe and decreases the cardiopulmonary changes associated with laparoscopic cholecystectomy.(2) Abdominal wall lift does not seem to offer advantage over pneumoperitoneum in any of the patient-oriented outcomes for laparoscopic cholecystectomy in patients with low anaesthetic risk and may increase costs by increasing the operating time. Hence it cannot be recommended routinely. More research on the topic is needed. PMID- 18425956 TI - Culture-specific programs for children and adults from minority groups who have asthma. AB - BACKGROUND: People with asthma who come from minority groups have poorer asthma outcomes and more asthma related visits to Emergency Departments (ED). Various programmes are used to educate and empower people with asthma and these have previously been shown to improve certain asthma outcomes. Models of care for chronic diseases in minority groups usually include a focus of the cultural context of the individual and not just the symptoms of the disease. Therefore, questions about whether culturally specific asthma education programmes for people from minority groups are effective at improving asthma outcomes, are feasible and are cost-effective need to be answered. OBJECTIVES: To determine whether culture-specific asthma programmes, in comparison to generic asthma education programmes or usual care, improve asthma related outcomes in children and adults with asthma who belong to minority groups. SEARCH STRATEGY: We searched the Cochrane Register of Controlled Trials (CENTRAL), the Cochrane Airways Group Specialised Register, MEDLINE, EMBASE, review articles and reference lists of relevant articles. The latest search was performed in March 2007. SELECTION CRITERIA: All randomised controlled trials (RCTs) comparing the use of culture-specific asthma education programmes with generic asthma education programmes, or usual care, in adults or children from minority groups who suffer from asthma. DATA COLLECTION AND ANALYSIS: Two review authors independently selected, extracted and assessed the data for inclusion. We contacted authors for further information if required. MAIN RESULTS: Three studies were eligible for inclusion in the review. A total of 396 patients, aged from 7 to 59 years were included in the meta-analysis of data. Use of a culture-specific programme was superior to generic programmes or usual care, in improving asthma quality of life scores in adults, pooled WMD 0.25 (95% CI 0.09 to 0.41) and asthma knowledge scores in children, WMD 3.30 (95% CI 1.07 to 5.53). There was no significant difference between groups in occurrence of asthma exacerbations, but the width of the confidence interval means that effects on exacerbation rates cannot be ruled out, rate ratio 0.93 (95% CI 0.80 to 1.10). AUTHORS' CONCLUSIONS: Culture specific programmes for adults and children from minority groups with asthma, have been found to be more effective than generic programmes in improving some (Quality of Life and asthma knowledge) but not all asthma outcomes. This evidence is limited by the small number of included studies and the lack of reported outcomes. Further trials are required to answer this question conclusively. PMID- 18425957 TI - Laparoscopic entry techniques. AB - BACKGROUND: Laparoscopy is a very common procedure in gynaecology. Complications associated with laparoscopy are often related to entry. The life-threatening complications include injury to the bowel, bladder, major abdominal vessels, and anterior abdominal-wall vessel. Other less serious complications can also occur, such as post-operative infection, subcutaneous emphysema and extraperitoneal insufflation. There is no clear consensus as to the optimal method of entry into the peritoneal cavity. OBJECTIVES: The objective of this study was to compare the different laparoscopic entry techniques in terms of their influence on intra operative and post-operative complications. SEARCH STRATEGY: This review has drawn on the search strategy developed by the Menstrual Disorders and Subfertility Group. In addition MEDLINE and EMBASE were searched through to July, 2007. SELECTION CRITERIA: Randomised controlled trials were included when one laparoscopic primary-port-entry technique was compared with another. DATA COLLECTION AND ANALYSIS: Data were extracted independently by the first two authors. Differences of opinion were registered and resolved by the fourth author. Results for each study were expressed as odds ratio (Peto version) with their 95% confidence intervals. MAIN RESULTS: The 17 included randomised controlled trials concerned 3,040 individuals undergoing laparoscopy. Overall there was no evidence of advantage using any single technique in terms of preventing major complications. However, there were two advantages with direct trocar entry when compared with Veress-Needle entry, in terms of avoiding extraperitoneal insufflation (OR 0.06, 95%CI 0.02, 0.23) and failed entry (OR 0.22, 95%CI 0.08, 0.56). There was also an advantage with radially expanding access system (STEP) trocar entry when compared with standard trocar entry, in terms of trocar site bleeding (OR 0.06, 95%CI 0.01, 0.46). Finally, there was an advantage of not lifting the abdominal wall before Veress-Needle insertion when compared to lifting in terms of failed entry without an increase in the complication rate (OR 5.17, 95%CI 2.24, 11.90). However, studies were limited to small numbers, excluding many patients with previous abdominal surgery and women with a raised body mass index, who often had unusually high complication rates. AUTHORS' CONCLUSIONS: On the basis of evidence investigated in this review, there appears to be no evidence of benefit in terms of safety of one technique over another. However, the included studies are small and cannot be used to confirm safety of any particular technique. PMID- 18425958 TI - Clinical pathways for chronic cough in children. AB - BACKGROUND: Chronic cough (a cough lasting longer than 4 weeks) is a common symptom presenting to primary care in Australia and internationally. Chronic cough costs the community, is distressing to parents, and ignoring cough may lead to delayed diagnosis and illness progression of serious underlying respiratory disease. Clinical guidelines have been shown to provide more efficient and effective patient care and can clarify clinical decision making. Cough guidelines have been designed to facilitate management of chronic cough, however treatment recommendations vary and specific clinical pathways for the treatment of chronic cough in children are important, as the cause and treatments for cough in a child vary significantly adults. Therefore, it would be beneficial to clinical practice to systematically evaluate the use of clinical pathways for the treatment of chronic cough in children. OBJECTIVES: To evaluate the effectiveness of using a clinical pathway in the management of children with chronic cough. SEARCH STRATEGY: The Cochrane Register of controlled Trials (CENTRAL), the Cochrane Airways Group Specialised Register, MEDLINE, EMBASE, review articles and reference lists of relevant articles were searched. The search was carried out in May 2007. SELECTION CRITERIA: All randomised controlled trials with parallel group design comparing use vs non-use of a clinical pathway for treatment of chronic cough in children. DATA COLLECTION AND ANALYSIS: Results of searches were reviewed against the pre-determined criteria for inclusion. Two reviewers independently selected the studies and it was planned that data extraction would have been done in duplicate. MAIN RESULTS: The search identified 471 potentially relevant titles but no studies met criteria for inclusion in the review. AUTHORS' CONCLUSIONS: Without further available evidence, recommendations for the use of clinical pathways for the treatment of chronic cough in children cannot be made. Until further evidence is available, the decision for further investigation and treatment for the child presenting with chronic cough should be made on an individual basis (i.e. dependent on symptoms and signs) with consideration for existing data from other Cochrane reviews on specific treatments for cough. Trials are required to provide evidence on the effectiveness of clinical pathways for the treatment of chronic cough in children. PMID- 18425959 TI - Anticoagulation for the long term treatment of venous thromboembolism in patients with cancer. AB - BACKGROUND: Cancer increases the risk of thromboembolic events and the risk of recurrent thromboembolic events while on anticoagulation. OBJECTIVES: To compare the efficacy and safety of low molecular weight heparin (LMWH) and oral anticoagulants (vitamin K antagonist (VKA) and ximelagatran) for the long term treatment of venous thromboembolism (VTE) in patients with cancer. SEARCH STRATEGY: A comprehensive search was undertaken including a January 2007 search of electronic databases; Cochrane Central Register of Controlled Trials (CENTRAL), (The Cochrane Library 2007, Issue 1). MEDLINE (1966 onwards; accessed via OVID), EMBASE (1980 onwards; accessed via OVID) and ISI the Web of Science. Hand search of the proceedings of the American Society of Clinical Oncology and of the American Society of Hematology. Checking of references of included studies, relevant papers and related systematic reviews. Use of "related article" feature in PubMed; and (5) search of ISI the Web of Science for papers citing landmark studies. SELECTION CRITERIA: Randomized controlled trials (RCTs) comparing long term treatment with LMWH versus oral anticoagulants (VKA or ximelagatran) in patients with cancer and symptomatic objectively confirmed VTE. DATA COLLECTION AND ANALYSIS: Using a standardized data form we extracted data on methodological quality, participants, interventions and outcomes of interest: survival, recurrent VTE, major bleeding, minor bleeding, thrombocytopenia and postphlebitic syndrome. MAIN RESULTS: Of 3986 identified citations, eight RCTs were eligible and reported data for patients with cancer. Their overall methodological quality was moderate. Meta-analysis of six RCTs showed that LMWH, compared to VKA provided no statistically significant survival benefit (Hazard ratio (HR) = 0.96; 95% CI 0.81 to 1.14) but a statistically significant reduction in VTE (HR = 0.47; 95% (Confidence Interval (CI) = 0.32 to 0.71). There was no statistically significant difference between LMWH and VKA in bleeding outcomes (RR = 0.91; 95% CI = 0.64 to 1.31) or thrombocytopenia (RR = 1.02; 95% CI = 0.60 to 1.74). One RCT compared tinzaparin and dalteparin and showed no differences in the outcomes of interest. One RCT compared a six months extension of anticoagulation with 18 months Ximelagatran 24mg twice daily versus placebo. It showed a reduction in VTE (HR = 0.16; 95% CI 0.09 to 0.30) with no apparent effect on survival or bleeding. AUTHORS' CONCLUSIONS: For the long term treatment of VTE in patients with cancer, LMWH compared to VKA reduces venous thromboembolic events but not death. The decision for a patient with cancer and VTE to start long term LMWH versus oral anticoagulation should balance the benefits and downsides and integrate the patient's values and preferences for the important outcomes and alternative management strategies. PMID- 18425960 TI - Vitamin C for preventing and treating tetanus. AB - BACKGROUND: Tetanus is a severe infection that can be prevented by vaccination. In developing countries vaccination coverage is not always high and in developed countries cases may still occur, particularly in elderly people owing to their reduced immunoprotection. It has been estimated that there are about one million cases of tetanus per year globally. In animal studies, vitamin C protected against various infections. In a study with rats, vitamin C protected against tetanus toxin. OBJECTIVES: To assess the prophylactic and therapeutic effect of vitamin C in tetanus. SEARCH STRATEGY: We searched the Cochrane Central Register of Controlled Trials (CENTRAL) (The Cochrane Library, 2007, issue 4), MEDLINE (1950 to January 2008), EMBASE (1980 to 2008 Week 03), the Cochrane Wounds Group Specialised Register (January 2008), the Cochrane Infectious Diseases Group Specialised Register (June 2007), and the reference lists of relevant reviews and monographs. SELECTION CRITERIA: We included controlled trials of vitamin C as a prevention or treatment for tetanus, whether or not placebo controlled, in any language, published or unpublished. Two authors independently made inclusion decisions. DATA COLLECTION AND ANALYSIS: Both review authors independently extracted data from trial reports. MAIN RESULTS: One single trial was eligible for inclusion. This non randomised, controlled, unblinded treatment trial involved 117 tetanus patients and was undertaken in Bangladesh. Vitamin C at a dosage of 1 g/day was administered intravenously alongside conventional treatment. At recruitment, the participants were stratified into two age groups and the results were reported by age. In the children aged 1 to 12 years (n = 62), vitamin C treatment was associated with a 100% reduction in tetanus mortality (95% confidence interval from -100% to -94%). In people aged 13 to 30 years (n = 55), vitamin C treatment was associated with a 45% reduction in tetanus mortality (95% confidence interval from -69% to -5%). AUTHORS' CONCLUSIONS: A single, non randomised, poorly reported trial of vitamin C as a treatment for tetanus suggests a considerable reduction in mortality. However, concerns about trial quality mean that this result must be interpreted with caution and vitamin C cannot be recommended as a treatment for tetanus on the basis of this evidence. New trials should be carried out to examine the effect of vitamin C on tetanus treatment. PMID- 18425961 TI - Dietary advice in pregnancy for preventing gestational diabetes mellitus. AB - BACKGROUND: Gestational diabetes mellitus (GDM) is a form of diabetes that occurs during pregnancy which can result in significant adverse outcomes for mother and child both in the short and long term. The potential for adverse outcomes, in addition to the increasing prevalence of gestational diabetes worldwide, demonstrates the need to assess strategies, such as dietary advice, that might prevent gestational diabetes. OBJECTIVES: To assess the effects of dietary advice in preventing gestational diabetes mellitus. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group's Trials Register (January 2008) and reference lists of retrieved articles. SELECTION CRITERIA: Quasi-randomised and randomised studies of dietary intervention for preventing glucose intolerance in pregnancy. DATA COLLECTION AND ANALYSIS: Two review authors independently conducted data extraction and quality assessment. We resolved disagreements through discussion or through a third author. MAIN RESULTS: Three trials (107 women) were included in the review. One trial (25 pregnant women) analysed high fibre diets with no included outcomes showing statistically significant differences. Two trials (82 pregnant women) assessed low glycaemic index (LGI) versus high glycaemic index diets for pregnant women. Women on the LGI diet had fewer large for gestational age infants (one trial; relative risk (RR) 0.09, 95% confidence interval (CI) 0.01 to 0.69), infants with lower ponderal indexes (two trials; weighted mean difference (WMD) -0.18, 95% CI -0.32 to -0.04, random effects analysis) and lower maternal fasting glucose levels (two trials; WMD 0.28 mmol/L 95% CI -0.54 to -0.02, random-effects model). Results for women on the LGI diet on neonatal birth weight were not conclusive under a random-effects model (two trials; WMD -527.64 g, 95% CI -1119.20 to 63.92); however, on a fixed effect model, women on the LGI diet gave birth to lighter babies (two trials; WMD -445.55 g, 95% CI -634.16 to -256.95). High heterogeneity was observed between the trials in most results and both were relatively small trials. One of these trials also included a standard exercise regimen for all participants. AUTHORS' CONCLUSIONS: While a low glycaemic index diet was seen to be beneficial for some outcomes for both mother and child, results from the review were inconclusive. Further trials with large sample sizes and longer follow up are required to make more definitive conclusions. No conclusions could be drawn from the high-fibre versus control-diet comparison since the trial involved did not report on many of the outcomes we prespecified. PMID- 18425962 TI - Locomotor training for walking after spinal cord injury. AB - BACKGROUND: Locomotor training for walking is used in rehabilitation after spinal cord injury (SCI) and might help to improve walking. OBJECTIVES: To assess the effects of locomotor training on improvement in walking for people with traumatic SCI. SEARCH STRATEGY: We searched the Cochrane Injuries Group Specialised Register (last searched June 2007); the Cochrane Central Register of Controlled Trials (CENTRAL) (The Cochrane Library 2007, Issue 2); MEDLINE (1966 to June 2007); EMBASE (1980 to June 2007); National Research Register (2007, Issue 2); CINAHL (1982 to June 2007); AMED (Allied and Complementary Medicine Database) (1985 to June 2007); SPORTDiscus (1949 to June 2007); PEDro (the Physiotherapy Evidence database) (searched June 2007); COMPENDEX (engineering databases) (1972 to June 2007); INSPEC (1969 to June 2007); and the National Research Register, Zetoc, and Current Controlled Trials research and trials registers. We also handsearched relevant conference proceedings, checked reference lists and contacted study authors in an effort to identify published, unpublished and ongoing trials. SELECTION CRITERIA: We included randomised controlled trials (RCT) that compared locomotor training to any other exercise provided with the goal of improving walking function after SCI or to a no-treatment control group. DATA COLLECTION AND ANALYSIS: Two review authors independently selected trials for inclusion, assessed trial quality and extracted the data. The primary outcomes were the speed of walking and walking capacity at follow up. MAIN RESULTS: Four RCTs involving 222 patients were included in this review. Overall, the results were inconclusive. There was no statistically significant effect of locomotor training on walking function after SCI comparing bodyweight supported treadmill training with or without functional electrical stimulation or robotic assisted locomotor training. AUTHORS' CONCLUSIONS: There is insufficient evidence from RCTs to conclude that any one locomotor training strategy improves walking function more than another for people with SCI. Research in the form of large RCTs is needed to address specific questions about the type of locomotor training which might be most effective in improving walking function of people with SCI. PMID- 18425963 TI - Duration of intravenous antibiotic therapy in people with cystic fibrosis. AB - BACKGROUND: Respiratory disease is the major cause of mortality and morbidity in cystic fibrosis (CF). Life expectancy of people with CF has increased dramatically in the last 40 years. One of the major reasons for this increase is the mounting use of antibiotics to treat chest exacerbations caused by bacterial infections. The optimal duration of intravenous antibiotic therapy is not clearly defined. Individuals usually receive intravenous antibiotics for 14 days, but treatment may range from 10 to 21 days. A shorter duration of antibiotic treatment risks inadequate clearance of infection which could lead to further lung damage. Prolonged courses of intravenous antibiotics are expensive and inconvenient and the incidence of allergic reactions to antibiotics also increases with prolonged courses. The use of aminoglycosides requires frequent monitoring to avoid some of their side effects. However, some organisms which infect people with CF are known to be multi-resistant to antibiotics, and may require a longer course of treatment. OBJECTIVES: To assess the optimal duration of intravenous antibiotic therapy for treating chest exacerbations in people with cystic fibrosis. SEARCH STRATEGY: We searched the Cochrane Cystic Fibrosis and Genetic Disorders Group Trials Register which comprises references identified from comprehensive electronic database searches, handsearches of relevant journals, abstract books and conference proceedings. Most recent search of the Group's Cystic Fibrosis Trials Register: February 2008. SELECTION CRITERIA: Randomised and quasi-randomised controlled trials comparing different durations of intravenous antibiotic courses for acute respiratory exacerbations in people with CF, either with the same drugs at the same dosage, the same drugs at a different dosage or frequency or different antibiotics altogether, including studies with additional therapeutic agents. DATA COLLECTION AND ANALYSIS: No eligible trials were identified. MAIN RESULTS: No eligible trials were identified. AUTHORS' CONCLUSIONS: There are no clear guidelines on the optimum duration of intravenous antibiotic treatment. Duration of treatment is currently based on unit policies and response to treatment. Shorter duration of treatment should improve quality of life and compliance; result in a reduced incidence of drug reactions; and be less costly. However, this may not be sufficient to clear a chest infection and may result in an early recurrence of an exacerbation. This systematic review identifies the need for a multicentre, randomised controlled trial comparing different durations of intravenous antibiotic treatment as it has important clinical and financial implications. PMID- 18425964 TI - Oral vasodilators for primary Raynaud's phenomenon. AB - BACKGROUND: Many different drugs have been suggested for the symptomatic treatment of primary Raynaud's phenomenon. Apart from calcium channel blockers, which are considered the drugs of choice, the evidence of the effects of alternative pharmacological treatments is limited. OBJECTIVES: To assess the effects of various drugs with vasodilator actions on primary Raynaud's phenomenon. SEARCH STRATEGY: The Cochrane Peripheral Vascular Diseases Group searched their Specialised Register (last searched 24 July 2007), and the Cochrane Central Register of Controlled Trials (CENTRAL) (last searched Issue 3, 2007). In addition, we searched MEDLINE (January 1966 to July 2007), EMBASE (1980 to July 2007) and reference lists of relevant articles. We contacted pharmaceutical companies. There were no language restrictions. SELECTION CRITERIA: Randomised controlled trials evaluating the effects of oral formulations of any drug with vasodilator effects on subjective symptoms in primary Raynaud's phenomenon. Treatment with, or comparison with, calcium channel blockers was not assessed in this review. DATA COLLECTION AND ANALYSIS: Both authors assessed the trials for inclusion and their quality. One author (BV) extracted the data MS checked the results. Data extraction included adverse events. we contacted trial authors for missing data. MAIN RESULTS: Eight studies involving 290 participants were included. Two trials examined the effects of captopril, the rest were single trials on single drugs. All comparisons were with placebo. The methodological quality of most trials was poor. Enalapril was associated with a small increase in the frequency of attacks per week (difference in means 0.8; 95% CI 0.43 to 1.17). The difference between the intervention groups on a subjective improvement score was non-significant. There was a significant effect of buflomedil on the frequency of attacks per week (weighted mean difference (WMD) -8.8; 95% CI -17.55 to -0.09), but there was no evidence of effect on the severity score. The proportion with fewer attacks was significantly higher on moxisylyte than on placebo (relative risk (RR) 4.33; 95% CI 1.36 to 13.81). For captopril, beraprost, dazoxiben and ketanserin there was no evidence of an effect on the frequency, severity or duration of attacks. Beraprost and moxisylyte gave significantly more adverse effects than placebo. AUTHORS' CONCLUSIONS: Poor methodological quality, small sample sizes and the limited data available resulted in low precision of the statistical results and limited value of the overall results . The overall results show that there is no evidence for an effect of vasodilator drugs on primary Raynaud's phenomenon. PMID- 18425965 TI - Pentoxifylline for diabetic retinopathy. AB - BACKGROUND: There is increasing evidence that capillary occlusion plays an important part in the development of diabetic retinopathy. Disaggregants, such as pentoxyfilline may influence the outcome and progression of diabetic retinopathy, but no systematic review of the literature on its efficacy and safety has been published to examine this hypothesis. OBJECTIVES: The aim of the current research was to review the literature in a systematic way in order to assess the effects of pentoxyfilline for diabetic retinopathy in methodologically robust trials. The null hypothesis was that pentoxyfilline has no influence on the progression of diabetic retinopathy or blindness. SEARCH STRATEGY: A systematic search of electronic databases was carried out to identify publications. Relevant papers, written in any language, were accessed and assessed for data. SELECTION CRITERIA: Only randomized controlled clinical trials (RCTs) evaluating the effects of pentoxyfilline in the treatment of diabetic retinopathy were to be included. DATA COLLECTION AND ANALYSIS: Two authors independently assessed studies for inclusion criteria and for risk of bias. MAIN RESULTS: A total of 97 publications were identified by the electronic search and two authors checked the abstracts. Of these, 17 were identified as potentially relevant trials providing information about treatment of patients with diabetic retinopathy using pentoxyfilline and were read in full. Unfortunately, no publication fulfilled our inclusion criteria. AUTHORS' CONCLUSIONS: No sound research to date has examined the treatment of diabetic retinopathy with pentoxyfilline in such a way as to indicate whether this form of intervention has a significant impact on the natural history of this clinical condition. The potential role of this substance in the treatment of diabetic retinopathy remains open to debate, and it is suggested that future research focusing on patient-relevant outcomes takes the opportunity of addressing this important issue directly. PMID- 18425966 TI - Psychostimulants for depression. AB - BACKGROUND: Depression is common, disabling, costly and under-treated. There are problems in the current first-line drug treatment, antidepressants, for moderate or severe depression. There is a body of research that has evaluated the effect of psychostimulants (PS) in the treatment of depression. This has not been reviewed systematically. OBJECTIVES: To determine the effectiveness of PS in the treatment of depression and to assess adverse events associated with PS. SEARCH STRATEGY: Databases CCDANCTR-Studies and CCDANCTR-References were searched on 21/6/2006. Cochrane Central Register of Controlled Trials, MEDLINE, EMBASE, PsycInfo, AMED, CINAHL, Dissertation Abstracts and the National Health Service Research Register were searched. SELECTION CRITERIA: Randomised controlled trials (RCTs) assessing the effectiveness of PS were included. The trial population comprised adults of either sex with a diagnosis of depression. DATA COLLECTION AND ANALYSIS: Two review authors extracted the data independently and assessed trial quality. Meta-analysis was considered for trials with comparable key characteristics. The primary outcome was depression symptoms, based on a continuous outcome, using the standardised mean difference (SMD), or a dichotomous measure of clinical response, using odds ratios (OR), with 95% confidence intervals (CI). MAIN RESULTS: Twenty-four RCTs were identified. The overall quality of the trials was low. Five drugs were evaluated; dexamphetamine, methylphenidate, methylamphetamine, pemoline and modafinil. Modafinil was evaluated separately as its pharmacology is different to that of the other PS. PS were administered as a monotherapy, adjunct therapy, in oral or intravenous preparation and in comparison with a placebo or an active therapy. Most effects were measured in the short term (up to four weeks). Thirteen trials had some usable data for meta-analyses. Three trials (62 participants) demonstrated that oral PS, as a monotherapy, significantly reduced short term depressive symptoms in comparison with placebo (SMD -0.87, 95% CI -1.40, -0.33, with non-significant heterogeneity. A similar effect was found for fatigue. In the short term PS were acceptable and well tolerated. Tolerance and dependence were under evaluated. No statistically significant difference in depression symptoms was found between modafinil and placebo. AUTHORS' CONCLUSIONS: There is some evidence that in the short-term, PS reduce symptoms of depression. Whilst this reduction is statistically significant, the clinical significance is less clear. Larger high quality trials with longer follow-up and evaluation of tolerance and dependence are needed to test the robustness of these findings and, furthermore, to explore which PS may be more beneficial and in which clinical situations they are optimal. PMID- 18425967 TI - Dipeptidyl peptidase-4 (DPP-4) inhibitors for type 2 diabetes mellitus. AB - BACKGROUND: In type 2 diabetes mellitus there is a progressive loss of beta-cell function. One new approach yielding promising results is the use of the orally active dipeptidyl peptidase-4 (DPP-4) inhibitors like sitagliptin and vildagliptin. OBJECTIVES: To assess the effects of dipeptidyl peptidase-4 (DPP-4) inhibitors for type 2 diabetes mellitus. SEARCH STRATEGY: Studies were obtained from computerised searches of MEDLINE, EMBASE and The Cochrane Library. SELECTION CRITERIA: Studies were included if they were randomised controlled trials in adult people with type 2 diabetes mellitus and had a trial duration of at least 12 weeks. DATA COLLECTION AND ANALYSIS: Two authors independently assessed risk of bias and extracted data. Pooling of studies was performed by means of fixed effect meta-analysis. MAIN RESULTS: Twenty-five studies of good quality were identified, 11 trials evaluated sitagliptin and 14 trials vildagliptin treatment. Altogether, 6743 patients were randomised in sitagliptin and 6121 patients in vildagliptin studies, respectively. Sitagliptin and vildagliptin studies ranged from 12 to 52 weeks duration. No data were published on mortality, diabetic complications, costs of treatment and health-related quality of life. Sitagliptin and vildagliptin therapy in comparison with placebo resulted in an HbA1c reduction of approximately 0.7% and 0.6%, respectively. Data on comparisons with active comparators were limited but indicated no improved metabolic control following DPP-4 intervention in contrast to other hypoglycaemic agents. Sitagliptin and vildagliptin therapy did not result in weight gain but weight loss was more pronounced following placebo interventions. No definite conclusions could be drawn from published data on sitagliptin and vildagliptin effects on measurements of beta-cell function. Overall, sitagliptin and vildagliptin were well tolerated, no severe hypoglycaemia was reported in patients taking sitagliptin or vildagliptin. All-cause infections increased significantly after sitagliptin treatment but did not reach statistical significance following vildagliptin therapy. All published randomised controlled trials of at least 12 weeks treatment with sitagliptin and vildagliptin only reported routine laboratory safety measurements AUTHORS' CONCLUSIONS: DPP-4 inhibitors have some theoretical advantages over existing therapies with oral antidiabetic compounds but should currently be restricted to individual patients. Long-term data especially on cardiovascular outcomes and safety are urgently needed before widespread use of these new agents. More information on the benefit-risk ratio of DPP-4 inhibitor treatment is necessary especially analysing adverse effects on parameters of immune function. Also, long-term data are needed investigating patient-oriented parameters like health-related quality of life, diabetic complications and all-cause mortality. PMID- 18425968 TI - Anticonvulsants for cocaine dependence. AB - BACKGROUND: Cocaine dependence is a major public health problem that is characterized by recidivism and a host of medical and psychosocial complications. Although effective pharmacotherapy is available for alcohol and heroin dependence none exists currently for cocaine dependence despite two decades of clinical trials primarily involving antidepressant, anti convulsivant and dopaminergic medications. There has been extensive consideration of optimal pharmacological approaches to the treatment of cocaine dependence with consideration of both dopamine antagonists and agonists. Anticonvulsants have been candidates for the treatment of addiction based on the hypothesis that seizure kindling-like mechanisms contribute to addiction. OBJECTIVES: To evaluate the efficacy and the acceptability of anticonvulsants for cocaine dependence SEARCH STRATEGY: We searched the Cochrane Drugs and Alcohol Groups specialised register (issue 4, 2007), MEDLINE (1966 - march 2007), EMBASE (1988 - march 2007), CINAHL (1982- to march 2007) SELECTION CRITERIA: All randomised controlled trials and controlled clinical trials which focus on the use of anticonvulsants medication for cocaine dependence DATA COLLECTION AND ANALYSIS: Two authors independently evaluated the papers, extracted data, rated methodological quality MAIN RESULTS: Fifteen studies (1066 participants) met the inclusion criteria for this review: the anticonvulsants drugs studied were carbamazepine, gabapentin, lamotrigine, phenytoin, tiagabine, topiramate, valproate. No significant differences were found for any of the efficacy measures comparing any anticonvulsants with placebo. Placebo was found to be superior to gabapentin in diminishing the number of dropouts, two studies, 81 participants, Relative Risk (RR) 3.56 (95% CI 1.07 to 11.82) and superior to phenythoin for side effects, two studies, 56 participants RR 2.12 (95% CI 1.08 to 4.17). All the other single comparisons are not statistically significant. AUTHORS' CONCLUSIONS: Although caution is needed when assessing results from a limited number of small clinical trials at present there is no current evidence supporting the clinical use of anticonvulsants medications in the treatment of cocaine dependence. Aiming to answer the urgent demand of clinicians, patients, families, and the community as a whole for an adequate treatment for cocaine dependence, we need to improve the primary research in the field of addictions in order to make the best possible use out of a single study and to investigate the efficacy of other pharmacological agent. PMID- 18425969 TI - Unfractionated or low-molecular weight heparin for induction of remission in ulcerative colitis. AB - BACKGROUND: There are a limited number of treatment options for patients with ulcerative colitis (UC). An increased risk of thrombosis in UC coupled with an observation that UC patients being treated with anticoagulant therapy for thrombotic events had an improvement in their bowel symptoms led to trials examining the use of unfractionated heparin (UFH) and low molecular weight heparins (LMWH) in patients with active UC. OBJECTIVES: To review randomized trials examining the efficacy of unfractionated heparin (UFH) or low molecular weight heparins (LMWH) for remission induction in patients with ulcerative colitis. SEARCH STRATEGY: The MEDLINE (PUBMED), and EMBASE databases, The Cochrane Central Register of Controlled Trials, the Cochrane IBD/FBD group specialized trials register, review papers on ulcerative colitis, and references from identified papers were searched in an effort to identify all randomized trials studying UFH or LMWH use in patients with ulcerative colitis. Abstracts from major gastroenterological meetings were searched to identify research published in abstract form only. SELECTION CRITERIA: Each author independently reviewed potentially relevant trials to determine their eligibility for inclusion based on the criteria identified above. The Jadad scale was used to assess study quality. Studies published in abstract form only were included if the authors could be contacted for further information. DATA COLLECTION AND ANALYSIS: A data extraction form was developed and used to extract data from included studies. At least 2 authors independently extracted data. Any disagreements were resolved by consensus. Data were analyzed using Review Manager (RevMan 4.2.9). Data were analyzed on an intention-to-treat basis, and treated dichotomously. In cross-over studies, only data from the first arm were included. The primary endpoint was induction of remission, as defined by the studies. Data were combined for analysis if they assessed the same treatments (UFH or LMWH versus placebo or other therapy). If a comparison was only assessed in a single trial, P-values were derived using the chi-square test. If the comparison was assessed in more than one trial, summary test statistics were derived using the Peto odds ratio and 95% confidence intervals (95% CI). The presence of heterogeneity among studies was assessed using the chi-square test (a P value of 0.10 was regarded as statistically significant). If statistically significant heterogeneity was identified the odds ratio and 95% CI were calculated using a random effects model. MAIN RESULTS: There were 2 randomized, double-blind studies assessing LMWH versus placebo for the treatment of mild-moderate active UC. Various outcomes were assessed in the 2 studies. LMWH showed no benefit over placebo in any outcome, including clinical remission (OR 1.09; 95% CI 0.26 to 4.63; P = 0.91), clinical improvement (OR 0.73; 95% CI 0.32 to 1.66; P = 0.45 and OR 1.09; 95% CI 0.18 to 6.58; P = 0.92 in the two studies, respectively), endoscopic improvement (OR 1.35; 95% CI 0.29 to 6.18; P = 0.70), or histological improvement (OR 2.00; 95% CI 0.45 to 8.96; P = 0.37). LMWH was also not beneficial when added to standard therapy in a randomized open-label trial in which the outcome measures included clinical remission (OR 0.71; 95% CI 0.17 to 2.95; P = 0.64), clinical improvement (OR 2.00; 95% CI 0.31 to 12.75; P = 0.46), endoscopic remission (OR 0.71; 95% CI 0.17 to 2.95; P = 0.64), or endoscopic improvement (OR 1.40; 95% CI 0.34 to 5.79; P = 0.64). LMWH was well-tolerated and provided no significant benefit for quality of life. One study examining UFH versus corticosteroids in the treatment of severe UC demonstrated inferiority of UFH in clinical improvement as an outcome measure (OR 0.02; 95% CI 0 to 0.40; P = 0.01). Patients assigned to UFH did not improve clinically. More patients assigned to UFH had rectal hemorrhage as an adverse event. AUTHORS' CONCLUSIONS: There is no evidence to support the use of UFH or LMWH for the treatment of active UC. No further trials examining these drugs for patients with UC are warranted, except perhaps a trial of UFH in patients with mild disease. Any benefit found would need to be weighed against a possible increased risk of rectal bleeding in patients with active UC. PMID- 18425970 TI - Traditional corticosteroids for induction of remission in Crohn's disease. AB - BACKGROUND: Historically, corticosteroids have been the most commonly used class of medication for induction of remission in Crohn's disease (CD). Corticosteroids down regulate production of inflammatory cytokines and interfere with NF-kappaB production, thereby blunting inflammatory response. OBJECTIVES: The primary objective was to systematically review the efficacy and safety of traditional corticosteroids (given orally or intravenously) for induction of remission in CD. SEARCH STRATEGY: The following electronic databases were searched: MEDLINE, EMBASE, the Cochrane Central Register of Controlled Trials, the Cochrane Inflammatory Bowel Disease and Functional Bowel Disorders (IBD/FBD) Group Specialized Trial Register, and ClinicalTrials.gov. No language restrictions were applied. Reference lists of trials and review articles, as well as recent proceedings from major gastroenterology meetings were manually searched. SELECTION CRITERIA: Randomized, controlled clinical trials of traditional, systemic corticosteroids for the induction of remission of active CD were included in this review. Control groups included patients receiving either placebo or 5-aminosalicylates (5-ASA). The study population included patients of any age with active CD (as defined by the study authors or validated clinical activity indices), receiving any formulation of systemically available corticosteroid by any oral or parenteral methods of delivery. The primary outcome was induction of remission of CD. Secondary outcomes included clinical response, change in mean CDAI, adverse events and the proportion of patients withdrawing due to adverse events. DATA COLLECTION AND ANALYSIS: Two independent investigators reviewed studies for eligibility, extracted the data and assessed study quality using Jadad's criteria. A random or fixed effects model was chosen based on an assessment of heterogeneity, and studies were weighted using the DerSimonian & Laird or the Mantel-Haenszel method accordingly. Meta-analysis was performed using RevMan 4.2.10 software. MAIN RESULTS: Two studies compared corticosteroids to placebo and six studies compared corticosteroids to 5-ASA. Corticosteroids were found to be significantly more effective than placebo at inducing remission in CD (RR 1.99; 95% CI 1.51 to 2.64; P < 0.00001). Corticosteroids were found to be more effective than 5-ASA at inducing remission in studies with long follow-up duration (i.e. > 15 weeks; RR 1.65; 95% CI 1.33 to 2.03; P < 0.00001). Corticosteroids induced adverse events in a higher proportion of patients than placebo (RR 4.89; 95% CI 1.98 to 12.07; P = 0.0006), or low-dose 5-ASA (RR 2.38; 95% CI 1.34 to 4.25; P = 0.003). No difference existed in the proportion of patients experiencing adverse events when steroids were compared to high-dose 5-ASA. Steroids did not induce more study withdrawals due to adverse events than either placebo or 5-ASA. AUTHORS' CONCLUSIONS: Corticosteroids are effective for induction of remission in patients with CD, particularly when used for more than 15 weeks. Although corticosteroids cause more adverse events than either placebo or low-dose 5-ASA, these adverse events did not lead to increased study withdrawal in the included studies. Further information is required to determine the optimal duration of treatment and tapering protocol to maximize the efficacy of treatment with corticosteroids. Additionally, further study is required to determine whether corticosteroids are more effective in patients with certain phenotypes or when administered intravenously. PMID- 18425971 TI - Allopurinol for preventing mortality and morbidity in newborn infants with suspected hypoxic-ischaemic encephalopathy. AB - BACKGROUND: Delayed neuronal death following a perinatal hypoxic insult is due partly to xanthine oxidase-mediated production of cytotoxic free radicals. Evidence exists that allopurinol, a xanthine-oxidase inhibitor, reduces delayed cell death in animal models of perinatal asphyxia and in human patients with other forms of organ reperfusion injury. OBJECTIVES: To determine the effect of allopurinol on mortality and morbidity in newborn infants with suspected hypoxic ischaemic encephalopathy. SEARCH STRATEGY: The standard search strategy of the Cochrane Neonatal Review Group was used. This included searches of the Cochrane Central Register of Controlled Trials (CENTRAL, The Cochrane Library, Issue 4, 2007), MEDLINE (1966 - December 2007), EMBASE (1980 - December 2007), conference proceedings, and previous reviews. SELECTION CRITERIA: Randomised or quasi randomised controlled trials that compared allopurinol administration vs. placebo or no drug in newborn infants with suspected hypoxic-ischaemic encephalopathy. DATA COLLECTION AND ANALYSIS: The standard methods of the Cochrane Neonatal Review Group were used, with separate evaluation of trial quality and data extraction by two authors. Data were synthesised using a fixed effects model and reported using typical relative risk, typical risk difference and weighted mean difference. MAIN RESULTS: Three trials in which a total of 114 infants participated were identified. In one trial, participants were exclusively infants with severe encephalopathy. The other trials also included infants with mild and moderately-severe encephalopathy. These studies were generally of good methodological quality, but were underpowered to detect clinically important effects of allopurinol on mortality and morbidity. Meta-analysis did not reveal a statistically significant difference in the risk of death during infancy [typical relative risk 0.92 (95% confidence interval 0.59 to 1.45); typical risk difference -0.03 (95% confidence interval -0.16 to 0.11)], nor in the incidence of neonatal seizures [typical relative risk 0.93 (95% confidence interval 0.75 to 1.16); typical risk difference -0.05 (95% confidence interval -0.21 to 0.11)]. Only one trial assessed neurodevelopment in surviving children and did not find a statistically significant effect. AUTHORS' CONCLUSIONS: The available data are not sufficient to determine whether allopurinol has clinically important benefits for newborn infants with hypoxic-ischaemic encephalopathy and, therefore, larger trials are needed. Such trials could assess allopurinol as an adjunct to therapeutic hypothermia in infants with moderate and severe encephalopathy and should be designed to exclude clinically important effects on mortality and adverse long-term neurodevelopmental outcomes. PMID- 18425972 TI - Bone grafts and bone substitutes for treating distal radial fractures in adults. AB - BACKGROUND: Surgical treatment of fractures of the distal radius can involve the implantation of bone scaffolding materials (bone grafts and substitutes) into bony defects that frequently arise after fracture reduction. OBJECTIVES: To review the evidence from randomised controlled trials evaluating the implanting of bone scaffolding materials for treating distal radial fractures in adults. SEARCH STRATEGY: We searched the Cochrane Bone, Joint and Muscle Trauma Group Specialised Register (June 2007), the Cochrane Central Register of Controlled Trials, MEDLINE, EMBASE and other databases, conference proceedings and reference lists. No language restrictions were applied. SELECTION CRITERIA: Randomised or quasi-randomised controlled clinical trials evaluating the use of bone scaffolding for treating distal radial fracture in adults. DATA COLLECTION AND ANALYSIS: Two people independently selected studies and undertook assessment and data collection. MAIN RESULTS: Ten heterogenous trials involving 874 adults with generally unstable fractures were grouped into six comparisons. No trial had proven allocation concealment. Four trials (239 participants) found implantation of bone scaffolding (autogenous bone graft (one trial); Norian SRS - a bone substitute (two trials); methylmethacrylate cement (one trial)) improved anatomical outcomes compared with plaster cast alone; and two found it improved function. Reported complications of bone scaffolding were transient discomfort resulting from extraosseous deposits of Norian SRS; with surgical removal of one intra-articular deposit. One trial (323 participants) comparing bone substitute (Norian SRS) versus plaster cast or external fixation found no difference in functional or anatomical outcomes at one year. Statistically significant complications in the respective groups were extraosseous Norian SRS deposits and pin track infection. One trial (48 participants with external fixation) found that autogenous bone graft did not significantly change outcome. There was one serious donor-site complication. One trial (21 participants) found some indication of worse outcomes for hydroxyapatite bone cement compared with Kapandji's intrafocal pinning. Three trials (180 participants) found bone scaffolding (autogenous bone graft (one trial); Norian SRS (one trial); methylmethacrylate cement (one trial)) gave no significant difference in functional outcomes but some indication of better anatomical outcomes compared with external fixation. Most reported complications were associated with external fixation; extraosseous deposits of Norian SRS occurred in one trial. One trial (93 participants with dorsal plate fixation) found autografts slightly improved wrist function compared with allogenic bone material but with an excess of donor site complications. AUTHORS' CONCLUSIONS: Bone scaffolding may improve anatomical outcome compared with plaster cast alone but there is insufficient evidence to conclude on functional outcome and safety; or for other comparisons. PMID- 18425973 TI - Personal assistance for adults (19-64) with both physical and intellectual impairments. AB - BACKGROUND: There is a high incidence of impairments among working age adults, and their prevalence is increasing in the West. Many countries offer personal assistance in the form of individualised support for people living in the community by a paid assistant other than a healthcare professional for at least 20 hours per week. OBJECTIVES: To assess the effectiveness of personal assistance for adults with physical and intellectual impairments, and the impacts of personal assistance on others, compared to other interventions. SEARCH STRATEGY: Electronic databases including CENTRAL, MEDLINE, EMBASE, CINAHL, PsycINFO, ERIC, Dissertation Abstracts International and a variety of specialist Swedish databases were searched from 1980 to June 2005; reference lists were checked; 345 experts, organisations, government bodies and charities were contacted in an attempt to locate relevant research. SELECTION CRITERIA: Adults (19-64) with permanent physical and intellectual impairments living in the community who require assistance to perform tasks of daily living (e.g., bathing and eating) and participate in normal activities. Controlled studies of personal assistance in which participants were prospectively assigned to study groups and in which control group outcomes were measured concurrently with intervention group outcomes were included. DATA COLLECTION AND ANALYSIS: Titles and abstracts were examined by two reviewers. Outcome data were extracted. Because no two studies made the same comparison, studies were not combined for meta-analyses. Studies were assessed for bias. Results and potential sources of bias are presented for included studies. MAIN RESULTS: Two studies involving 1002 participants compared personal assistance versus usual care. Whilst personal assistance was generally preferred over other services, some people prefer other services. Personal assistance may have some benefits for some recipients and may benefit caregivers. Paid assistance probably substitutes for informal care and may cost government more than alternatives; however, some evidence suggests it may reduce costs. The total costs to recipients and society are unknown. AUTHORS' CONCLUSIONS: Research in this field is limited. Personal assistance is expensive and difficult to organise, especially in places that do not already have services in place, but its total cost relative to other services is unknown. When implementing new programmes, recipients could be randomly assigned to different forms of assistance (e.g. organised by individual users versus organised through a cooperative). While advocates may support personal assistance for myriad reasons, this review demonstrates that further studies are required to determine which models of assistance are most effective and efficient for particular people. PMID- 18425974 TI - Strategies for communicating contraceptive effectiveness. AB - BACKGROUND: Knowledge of contraceptive effectiveness is crucial to making an informed choice. The consumer has to comprehend the pros and cons of the contraceptive methods being considered. Choice may be influenced by understanding the likelihood of pregnancy with each method and factors that influence effectiveness. OBJECTIVES: To review all randomized controlled trials comparing strategies for communicating to consumers the effectiveness of contraceptives in preventing pregnancy. SEARCH STRATEGY: We searched the computerized databases MEDLINE, POPLINE, CENTRAL, PsycINFO, and EMBASE for studies of communicating contraceptive effectiveness. We also examined references lists of relevant articles, and wrote to known investigators for information about other published or unpublished trials. SELECTION CRITERIA: We included randomized controlled trials that compared methods for communicating contraceptive effectiveness to consumers. The comparison could be usual practice or an alternative to the experimental intervention. DATA COLLECTION AND ANALYSIS: Data were abstracted by two authors and entered into RevMan. For dichotomous variables, the Peto odds ratio (OR) with 95% confidence intervals (CI) was calculated. For continuous variables, the weighted mean difference (WMD) was computed. MAIN RESULTS: Five trials met the inclusion criteria. In one study, knowledge gain favored a slide and-sound presentation versus a physician's oral presentation (WMD -19.00; 95% CI -27.52 to -10.48). Another trial showed a table with effectiveness categories led to more correct answers than one based on numbers [ORs were 2.42 (95% CI 1.43 to 4.12) and 2.19 (95% CI 1.21 to 3.97)] or a table with categories and numbers [ORs were 2.58 (95% CI 1.5 to 4.42) and 2.03 (95% CI 1.13 to 3.64)]. One trial examined contraceptive choice: women in the expanded program were more likely to choose sterilization (OR 4.26; 95% CI 2.46 to 7.37) or use a modern contraceptive method (OR 2.35; 95% CI 1.82 to 3.03). No trial had an explicit theoretical base, but each used concepts from common theories or models. AUTHORS' CONCLUSIONS: We have limited evidence about what works to help consumers choose an appropriate contraceptive method. For presenting pregnancy risk data, one trial showed that categories were better than numbers. In another trial, audiovisual aids worked better than the usual oral presentation. Strategies for communicating information should be examined in clinical settings and assessed for effect on contraceptive choice and retention of knowledge. To expand the knowledge base of what works in contraceptive counseling, randomized trials could intentionally use and test theories or models. PMID- 18425975 TI - Opportunities provision for preventing youth gang involvement for children and young people (7-16). AB - BACKGROUND: Youth gangs have long been studied in the United States and interest elsewhere is increasing. Many studies document a robust and consistent relationship between gang membership and elevated delinquency. One theory of gang involvement, drawing on anomie and strain theories, proposes that the gang provides a means of fulfilling the economic needs of youth excluded from legitimate labour markets. Opportunities provision is a gang prevention strategy based on this theory and the principle that providing youth with educational and employment opportunities may reduce gang involvement. Common techniques within opportunities provision include tutoring, remedial education, job training, and job placement. OBJECTIVES: To determine the effectiveness of opportunities provision for preventing youth gang involvement for children and young people aged 7 to 16. SEARCH STRATEGY: Electronic searches were conducted of ASSIA, CINAHL, CJA, Cochrane Library, Dissertations Abstracts, EMBASE, ERIC, IBSS, LILACs, LexisNexis Butterworths, MEDLINE, NCJR Service Abstracts Database, PsycINFO, and Sociological Abstracts, to April 2007. Reviewers contacted relevant organisations, individuals and list-servs and searched pertinent websites and reference lists. SELECTION CRITERIA: All randomised controlled trials or quasi randomised controlled trials of interventions that have opportunities provision as the majority component, delivered to children and youths aged 7 to 16 not involved in a gang, compared to any other or no intervention. DATA COLLECTION AND ANALYSIS: Searches yielded 2,696 unduplicated citations. 2,676 were excluded based on title and abstract. Two were excluded based on personal communication with study authors. Full-text reports for 18 citations were retrieved. 16 were excluded because they were not evaluations, did not address a gang prevention programme, did not include gang-related outcomes, did not include opportunities provision intervention components, or presented preliminary findings for outcomes reported in another citation. The remaining two reports were at least partially relevant to opportunities provision for gang prevention, but methodological flaws excluded both from analysis. MAIN RESULTS: No randomised controlled trials or quasi-randomised controlled trials were identified. AUTHORS' CONCLUSIONS: No evidence from randomised controlled trials or quasi-randomised controlled trials currently exists regarding the effectiveness of opportunities provision for gang prevention. Only two studies addressed opportunities provision as a gang prevention strategy, a case study and a qualitative study, both of which had such substantial methodological limitations that even speculative conclusions as to the impact of opportunities provision were impossible. Rigorous primary evaluations of gang prevention strategies are crucial to develop this research field, justify funding of existing interventions, and guide future gang prevention programmes and policies. PMID- 18425976 TI - Cognitive-behavioural interventions for preventing youth gang involvement for children and young people (7-16). AB - BACKGROUND: Many studies document a robust and consistent relationship between gang membership and elevated delinquency, with gang members disproportionately involved in crime compared to non-gang peers. Research also indicates that both delinquent youth and youth who join gangs often show a wide range of deficient or distorted social-cognitive processes compared to non-delinquent peers. Cognitive behavioural interventions are designed to address cognitive deficits in order to reduce maladaptive or dysfunctional behaviour, and studies have documented their positive impact on a number of behavioural and psychological disorders among children and youth. OBJECTIVES: To determine the effectiveness of cognitive behavioural interventions for preventing youth gang involvement for children and young people (ages 7-16). SEARCH STRATEGY: Electronic searches of ASSIA, CINAHL, CJA, Cochrane Library, Dissertations Abstracts A, EMBASE, ERIC, IBSS, LILACs, LexisNexis Butterworths, MEDLINE, NCJR Service Abstracts Database, PsycINFO, and Sociological Abstracts, to April 2007. Reviewers contacted relevant organisations, individuals, and list-servs and searched pertinent websites and reference lists. SELECTION CRITERIA: All randomised controlled trials or quasi randomised controlled trials of interventions with a cognitive-behavioural intervention as the majority component, delivered to youth and children aged 7-16 not involved in a gang. DATA COLLECTION AND ANALYSIS: Searching yielded 2,284 unduplicated citations, 2,271 of which were excluded as irrelevant based on title and abstract. One was excluded following personal communication with investigators. One citation, of a large randomised prevention trial, awaits assessment; personal communication with study authors yielded unpublished reports addressing gang outcomes, but insufficient detail precluded determining inclusion status. Seven remaining reports were excluded as irrelevant because they were narrative reviews or descriptions of programs without evaluations, did not address a gang prevention programme, or did not address a gang prevention program that included a cognitive-behavioural intervention. The remaining four full-text reports excluded because of study design, leading to 0 included studies. MAIN RESULTS: No randomised controlled trials or quasi-randomised controlled trials were identified. AUTHORS' CONCLUSIONS: No evidence from randomised controlled trials or quasi-randomised controlled trials exists regarding the effectiveness of cognitive-behavioural interventions for gang prevention. Four evaluations of Gang Resistance Education and Training (GREAT) have been conducted, two of which were part of a US national evaluation, but all were excluded based on study design. Reviewers conclude there is an urgent need for rigorous primary evaluations of cognitive-behavioural interventions for gang prevention to develop this research field and guide future gang prevention programmes and policies. PMID- 18425977 TI - Ciclesonide versus other inhaled steroids for chronic asthma in children and adults. AB - BACKGROUND: Inhaled corticosteroids (ICS) are an integral part of asthma management, and act as an anti-inflammatory agent in the airways of the lung. These agents confer both significant benefit in terms of symptom management and improvement in lung function, but may also cause harm in terms of local and systemic side-effects. Ciclesonide is a novel steroid that is metabolised to its active component in the lung, making it a potentially useful for reducing local side effects. OBJECTIVES: To assess the efficacy and adverse effects of ciclesonide relative to those of other inhaled corticosteroids in the management of chronic asthma. SEARCH STRATEGY: We searched the Cochrane Airways Group register of trials with pre-defined terms. Additional searches of PubMed and Clinicalstudyresults.org were undertaken. The literature searches for this review are current up to June 2007. SELECTION CRITERIA: Randomised parallel or crossover studies were eligible for the review. We included studies comparing ciclesonide with other steroids both at nominally equivalent dose or lower doses of ciclesonide. DATA COLLECTION AND ANALYSIS: Two review authors independently assessed trial quality and extracted data. Study authors were contacted for additional information. Adverse effects information was collected from the trials. MAIN RESULTS: Twenty one trials involving 7243 participants were included. Equal daily doses of ciclesonide and beclomethasone (BDP) or budesonide (BUD) gave similar results for peak expiratory flow rates (PEF), although forced vital capacity (FVC) was higher with ciclesonide. Data on forced expired volume in one second (FEV1) were inconsistent. Withdrawal data and symptoms were similar between treatments. Compared with the same dose of fluticasone (FP), data on lung function parameters (FEV1, FVC and PEF) did not differ significantly. Paediatric quality of life score favoured ciclesonide. Candidiasis was less frequent with ciclesonide, although other side-effect outcomes did not give significant differences in favour of either treatment. When lower doses of ciclesonide were compared to BDP or BUD, the difference in FEV1 did not reach significance but we cannot exclude a significant effect in favour of BDP/BUD. Other lung function outcomes did not give significant differences between treatments. Paediatric quality of life scores did not differ between treatments. Adverse events occurred with similar frequency between ciclesonide and BDP/BUD. Comparison with FP at half the nominal dose was undertaken in three studies, which indicated that FEV1 was not significantly different, but was not equivalent between the treatments (per protocol: -0.05 L 95% confidence intervals -0.11 to 0.01). AUTHORS' CONCLUSIONS: The results of this review give some support to ciclesonide as an equivalent therapy to other ICS at similar nominal doses. The studies assessed low doses of steroids, in patients whose asthma required treatment with low doses of steroids. At half the dose of FP and BDP/BUD, the effects of ciclesonide were more inconsistent The effect on candidiasis may be of importance to people who find this to be problematic. The role of ciclesonide in the management of asthma requires further study, especially in paediatric patients. Further assessment against FP at a dose ratio of 1:2 is a priority. PMID- 18425978 TI - Five-alpha-reductase Inhibitors for prostate cancer prevention. AB - BACKGROUND: Five-alpha-reductase inhibitors (5ARI) are frequently used to treat bothersome lower urinary tract symptoms associated with benign prostatic hyperplasia and male androgenic alopecia. They have potential as chemopreventive agents. OBJECTIVES: We sought to estimate the effectiveness and harms of 5ARI in preventing prostate cancer. SEARCH STRATEGY: MEDLINE, PreMEDLINE, and the Cochrane Collaboration Library were searched through April 2007 to identify randomized trials. SELECTION CRITERIA: For prostate cancer outcomes we included randomized controlled trials of at least 1 year in duration published after 1984. For non-prostate cancer outcomes, randomized trials were included if: they were at least 6 months in duration published after 1999. DATA COLLECTION AND ANALYSIS: The primary outcome was prostate cancer period-prevalence "for-cause." "For cause" was defined as prostate cancer clinically detected based on symptoms, an abnormal digital rectal exam, or detected as a result of an abnormal prostate specific antigen value. Trials were categorized as long (> 2 year), mid (1-2 years) and short (< 1 year) term. MAIN RESULTS: Nine trials reported prostate cancer period-prevalence. Three trials using finasteride lasted 4 years or longer but only one (the Prostate Cancer Prevention Trial) was specifically designed to assess the impact of 5ARI on prostate cancer period-prevalence. The mean age of enrollees was 64.6 years, 91% were white, mean PSA was 2.1 ng/mL. For-cause prostate cancers comprised 54% of all cancers detected. Finasteride was associated with a 26% relative risk reduction in prostate cancers detected for cause among all randomized subjects (relative risk 0.74 [95% CI 0.67 to 0.83]; absolute risk reduction = 1.4% (3.5% versus 4.9%). Six trials assessed prostate cancers detected overall with a pooled 26% relative reduction favoring 5ARI (relative risk 0.74 [95% CI 0.55 to1.00]; 2.9% absolute reduction (6.3% versus 9.2%). Reductions were observed regardless of age, race or family history of prostate cancer but not among men with baseline PSA > 4.0 ng/mL. A greater number of high Gleason score tumors (7-10 or 8-10) occurred in men on finasteride in the PCPT. Impaired sexual or erectile function or endocrine effects were more common with finasteride than placebo. AUTHORS' CONCLUSIONS: 5ARI reduce prostate cancer risk but may increase the risk of high-grade disease in men who are undergoing regular screening for prostate cancer using prostate specific antigen and digital rectal examination. Effects are consistent across race, family history and age and possibly 5ARI but were limited to men with baseline PSA values <4.0 ng/mL. The impact of 5ARI on absolute or relative rates of prostate cancer in men who are not being regularly screened is not clear. Information is inadequate to assess the impact of 5ARI on mortality. PMID- 18425979 TI - Adjuvant treatment of anaplastic oligodendrogliomas and oligoastrocytomas. AB - BACKGROUND: AO and AOA are known to be chemosensitive tumors. However, the impact of adding chemotherapy to surgery and radiotherapy has not been studied. Also, the value of chromosome 1p and 19q deletions as prognostic and predictive markers is only beginning to be defined. OBJECTIVES: After surgery, to compare radiotherapy (RT) plus chemotherapy versus RT alone (standard of care) in adults with newly diagnosed AO or mixed AOA. To investigate the prognostic and predictive value of loss of heterozygosity of chromosomes 1p and 19q. Outcomes analyzed include overall survival (OS), progression-free survival (PFS), and treatment toxicity greater than or equal to grade 3. SEARCH STRATEGY: Cochrane Central Register for Controlled Trials (CENTRAL, Issue 4,2006), MEDLINE (1966 to 2006) and EMBASE (1988 to 2006) were searched. Reference lists from relevant studies were scanned for any additional relevant articles. SELECTION CRITERIA: RCTs of adults with AO or mixed AOA comparing surgery plus RT versus surgery plus RT plus chemotherapy were included. No specific chemotherapy regimens were excluded. DATA COLLECTION AND ANALYSIS: Relevant studies were critically appraised and data was extracted. Based on the differences in patient selection with respect to the definition of AO (2 versus 3 high risk anaplastic features) and sequence of treatment (RT and chemotherapy), the results from the two RCTs were not able to be considered for meta-analysis. MAIN RESULTS: Two RCTs have tested surgery plus RT plus early procarbazine, lomustine, and vincristine (PCV) chemotherapy versus surgery plus RT alone. Neither study observed a survival benefit with the addition of early PCV chemotherapy. However, both studies found a statistically significant increase in PFS associated with the administration of PCV chemotherapy before surgery or after surgery and RT, with the benefit ranging from 10 to 11 months. Co-deletion of chromosomes 1p and 19q identifies a favorable subgroup of tumors with better overall survival outcomes. The predictive value of 1p and 19q co-deletions is less clear with one study observing a longer PFS with chemotherapy, while the other study did not. AUTHORS' CONCLUSIONS: Early PCV chemotherapy in addition to standard treatment of surgery and RT does not improve OS in patients with AO or AOA. However, it does improve PFS. It also is associated with significant toxicities. Tumors with 1p and 19q co deletions are associated with better OS and may indicate a more chemo-responsive tumor. PMID- 18425980 TI - Antioxidant supplements for prevention of mortality in healthy participants and patients with various diseases. AB - BACKGROUND: Animal and physiological research as well as observational studies suggest that antioxidant supplements may improve survival. OBJECTIVES: To assess the effect of antioxidant supplements on mortality in primary or secondary prevention randomised clinical trials. SEARCH STRATEGY: We searched The Cochrane Library (Issue 3, 2005), MEDLINE (1966 to October 2005), EMBASE (1985 to October 2005), and the Science Citation Index Expanded (1945 to October 2005). We scanned bibliographies of relevant publications and wrote to pharmaceutical companies for additional trials. SELECTION CRITERIA: We included all primary and secondary prevention randomised clinical trials on antioxidant supplements (beta-carotene, vitamin A, vitamin C, vitamin E, and selenium) versus placebo or no intervention. Included participants were either healthy (primary prevention trials) or had any disease (secondary prevention trials). DATA COLLECTION AND ANALYSIS: Three authors extracted data. Trials with adequate randomisation, blinding, and follow up were classified as having a low risk of bias. Random-effects and fixed-effect meta-analyses were performed. Random-effects meta-regression analyses were performed to assess sources of intertrial heterogeneity. MAIN RESULTS: Sixty seven randomised trials with 232,550 participants were included. Forty-seven trials including 180,938 participants had low risk of bias. Twenty-one trials included 164,439 healthy participants. Forty-six trials included 68111 participants with various diseases (gastrointestinal, cardiovascular, neurological, ocular, dermatological, rheumatoid, renal, endocrinological, or unspecified). Overall, the antioxidant supplements had no significant effect on mortality in a random-effects meta-analysis (relative risk [RR] 1.02, 95% confidence interval [CI] 0.99 to 1.06), but significantly increased mortality in a fixed-effect model (RR 1.04, 95% CI 1.02 to 1.06). In meta-regression analysis, the risk of bias and type of antioxidant supplement were the only significant predictors of intertrial heterogeneity. In the trials with a low risk of bias, the antioxidant supplements significantly increased mortality (RR 1.05, 95% CI 1.02 to 1.08). When the different antioxidants were assessed separately, analyses including trials with a low risk of bias and excluding selenium trials found significantly increased mortality by vitamin A (RR 1.16, 95% CI 1.10 to 1.24), beta-carotene (RR 1.07, 95% CI 1.02 to 1.11), and vitamin E (RR 1.04, 95% CI 1.01 to 1.07), but no significant detrimental effect of vitamin C (RR 1.06, 95% CI 0.94 to 1.20). Low-bias risk trials on selenium found no significant effect on mortality (RR 0.91, 95% CI 0.76 to 1.09). AUTHORS' CONCLUSIONS: We found no evidence to support antioxidant supplements for primary or secondary prevention. Vitamin A, beta-carotene, and vitamin E may increase mortality. Future randomised trials could evaluate the potential effects of vitamin C and selenium for primary and secondary prevention. Such trials should be closely monitored for potential harmful effects. Antioxidant supplements need to be considered medicinal products and should undergo sufficient evaluation before marketing. PMID- 18425983 TI - Half peak systolic velocity deceleration time: a new index for umbilical artery Doppler, but not a new Doppler parameter for waveform quantification. PMID- 18425981 TI - Physical characterization and in vivo evaluation of poloxamer-based DNA vaccine formulations. AB - BACKGROUND: Plasmid DNA (pDNA) vaccines have generated significant interest for the prevention or treatment of infectious diseases. Broader applications may benefit from the identification of safe and potent vaccine adjuvants. This report describes the development of a novel polymer-based formulation to enhance the immunogenicity of pDNA-based vaccines. METHODS: Plasmid DNA was formulated with a nonionic block copolymer, poloxamer CRL1005, and the cationic surfactant benzalkonium chloride (BAK) to produce a thermodynamically stable, self assembling system. The influence of parameters such as polymer concentration and BAK composition on the immune responses was evaluated in mice vaccinated with pDNA encoding influenza nucleoprotein. RESULTS: At concentrations of 7.5 mg/ml CRL1005, 0.3 mM BAK and 5 mg/ml pDNA, CRL1005/BAK/pDNA particles had a mean diameter of 261 +/- 0.2 nm and a surface charge of - 11.6 +/- 0.9 mV. The negative surface charge and atomic force microscopy images suggested that pDNA binds to BAK adsorbed to the surface of poloxamer particles. The CRL1005/BAK/pDNA formulation significantly enhanced antigen-specific cellular and humoral immune responses, and increased transgene levels in muscle and serum. The complexity of the formulation was reduced by replacing the commercial BAK, which is a mixture of four alkyl chains, with a C14 BAK homolog. The substitution yielded an analytically preferable formulation with equivalent physical characteristics and immunogenicity. CONCLUSIONS: The results suggest that the CRL1005/BAK/pDNA formulation may enhance immunogenicity by improving the delivery of pDNA-based vaccines. This formulation is currently being evaluated for the prevention of CMV associated disease in a phase 2 clinical trial. PMID- 18425985 TI - Long-term follow up of initial clinical cases with NF-kappaB decoy oligodeoxynucleotide transfection at the site of coronary stenting. AB - BACKGROUND: An essential transcription factor for inflammation, nuclear factor kappa B (NF-kappaB), plays a pivotal role for restenosis after percutaneous coronary intervention (PCI). To evaluate the safety and effectiveness of NF kappaB decoy oligodeoxynucleotides (ODN) to prevent restenosis, we initiated an open-label phase I/IIa clinical trial. METHODS: Seventeen patients who were suffering from angina with organic coronary stenosis were treated with NF-kappaB decoy ODN after PCI using bare metal stents. RESULTS: Although the average coronary stenosis was 90.8 +/- 7.0% before the stent implantation, the stenosis improved to 1.4 +/- 5.9% after the intervention. Serum MCP-1 levels were significantly suppressed in NF-kappaB decoy ODN treated patients compared to those in non-treated patients on day 3 after the PCI. Ticlopidine treatment was employed, since clopidogrel was not launched in Japan. Six months after the PCI and decoy ODN transfection, significant restenosis was found in only 1 of the 17 patients, and the average restenosis rate was 39.6 +/- 22.3%. No in-stent thrombosis was found and no significant systemic adverse effect occurred in any of the patients in this observation period. CONCLUSIONS: These results suggest the clinical usefulness and safety of the NF-kappaB decoy ODN transfer after PCI, although further placebo control trials are necessary. PMID- 18425987 TI - The safety of rosuvastatin in comparison with other statins in over 25,000 statin users in the Saskatchewan Health Databases. AB - PURPOSE: To compare mortality and the incidence of hospitalization for myopathy, rhabdomyolysis, acute renal failure and acute liver injury in patients receiving rosuvastatin and those taking other statins. METHODS: Patients prescribed a statin that they had not used before were selected from the Saskatchewan Health Databases (SHD) and followed up from 1 July 2003 until 31 March 2005. RESULTS: We studied 10,384 patients on rosuvastatin and 14,854 taking other statins. Two cases of myopathy were identified (one on rosuvastatin, one on another statin). The relative risk (RR) of myopathy in patients currently taking rosuvastatin compared with other statins was 1.31 (95% confidence interval [CI]: 0.13-13.41). Two cases of rhabdomyolysis were detected among current rosuvastatin users (incidence: 2.9 [95% CI: 0.8-10.7] per 10 000 person-years). No cases of acute liver injury occurred among rosuvastatin patients. Seventeen cases of acute renal failure were identified (five among rosuvastatin users, 12 taking other statins). The RR of acute renal failure in current rosuvastatin users compared with other statins was 0.49 (95% CI: 0.16-1.50). We identified 285 deaths during the study period (87 among rosuvastatin users, 198 taking other statins). The RR of death in current rosuvastatin users compared with other statins was 0.42 (95% CI: 0.32 0.57). CONCLUSIONS: We found no evidence that patients prescribed rosuvastatin were at greater risk of the study outcomes than patients prescribed other statins. There was no evidence of increased mortality among patients taking rosuvastatin, even after allowing for age, sex and prior statin use. PMID- 18425988 TI - The safety of rosuvastatin in comparison with other statins in over 100,000 statin users in UK primary care. AB - PURPOSE: To compare mortality and the incidence of hospitalization for myopathy, rhabdomyolysis, acute renal failure and acute liver injury in patients receiving rosuvastatin and those taking other statins. METHODS: Patients prescribed a statin that they had not used before were selected from the UK General Practice Research Database (GPRD) and followed up from 1 April 2003 to 31 December 2005. RESULTS: We studied 10 289 patients on rosuvastatin and 117 102 taking other statins. No cases of myopathy, rhabdomyolysis or acute liver injury occurred among rosuvastatin users. In those taking statins other than rosuvastatin, the incidence of myopathy was 0.4 (95% confidence interval (CI): 0.1-0.9), of rhabdomyolysis was 0.4 (95%CI: 0.1-0.9) and of acute liver injury was 0.4 (95%CI: 0.2-1.0), per 10 000 person-years. Fourteen cases of acute renal failure were identified (two among rosuvastatin users and 12 among other statin users). Among current users, the relative risk (RR) of acute renal failure in rosuvastatin users compared with other statin users was 1.16 (95%CI: 0.15-9.03).We identified 3232 deaths during the study period (173 in the rosuvastatin-treated group and 3059 in the other statin group). The RR of death associated with current use of rosuvastatin compared with other statins was 0.55 (95%CI: 0.44-0.68). CONCLUSIONS: We found no evidence that patients prescribed rosuvastatin were at greater risk of these outcomes than patients prescribed other statins. There was no evidence of increased mortality among patients taking rosuvastatin, even after allowing for age, sex and prior statin use. PMID- 18425989 TI - Reasons for psychiatric consultation referrals in Dutch nursing home patients with dementia: a comparison with normative data on prevalence of neuropsychiatric symptoms. AB - OBJECTIVE: To study psychiatric consultation referrals of nursing home patients with dementia and to compare referral reasons with normative data on prevalence of neuropsychiatric symptoms. METHODS: This is part of a cross-sectional study of 787 patients residing in 14 nursing homes in the Netherlands. Nursing home physicians (NHP) noted the primary reasons for psychiatric consultation according to the Neuropsychiatric Inventory items and two extra domains. Patients were subsequently assessed by an old-age psychiatrist. Eligible patients were those that had dementia. Reasons for referral were compared with independent data on prevalence of neuropsychiatric symptoms in nursing home patients with dementia. RESULTS: A total of 325/787 (41.3%) patients had dementia. Agitation, disinhibition and aberrant motor behaviour were frequent reasons for referral (>25%). Psychotic symptoms, apathy and eating behaviour changes were infrequent reasons (<10%) for seeking consultation. Agitation and disinhibition were more often primary reasons for consultation than would have been expected based on normative prevalence estimates of these symptoms. In contrast, delusions, euphoria, apathy, irritability and eating behaviour changes were less often reasons for referral compared with prevalence estimates. CONCLUSIONS: This study is the first to examine psychiatric consultation for dementia patients in Dutch nursing home. Large differences exist between referral reasons and normative data on symptom prevalence. Specialized mental health service was provided for the agitated and disinhibited patient in particular. Chances are that this is at the expense of the apathetic, retarded and quietly 'not causing any trouble' patient. PMID- 18425990 TI - Normative study of the Stroop Color and Word Test in an educationally diverse elderly population. AB - OBJECTIVE: The purpose of this study was to explore the effects of demographic variables on Stroop Color and Word Test (SCWT) performance in an educationally diverse elderly population and to provide normative information. METHODS: SCWT was administered to 564 community-dwelling volunteers aged 60-90 years with an educational history of from zero to 25 years of full-time education. People with serious neurological, medical and psychiatric disorders (including dementia) were excluded. RESULTS: Age, education and gender were found to be significantly associated with performance on all three pages of the SCWT. Based on the results obtained, SCWT norms were stratified by age (four overlapping tables), education (three strata), and gender. CONCLUSIONS: In the present study, normative information on SCWT was obtained from an educationally diverse elderly population. SCWT would appear to be more useful in poorly educated elderly, and could be used in future cross-cultural comparisons of geriatric populations. PMID- 18425992 TI - A lactoferrin-derived peptide with cationic residues concentrated in a region of its helical structure induces necrotic cell death in a leukemic cell line (HL 60). AB - Model studies have shown that peptides derived from the N-terminal region of bovine lactoferrin (Lf-B) exhibit antitumor activity against certain cell lines. This activity is due primarily to the peptides' apoptogenic effect. Several reports indicate that cationic residues clustered in two regions of the peptide sequence can be shuffled into one region and thereby increase cytotoxic activity, although the mechanism of this enhanced cytotoxic effect has not been clarified. In this paper, we considered several parameters that determine the mode of cell death after exposure to a native Lf-B derived peptide (Pep1, residues 17-34), and a modified peptide (mPep1) wherein the cationic residues of Pep1 are clustered in a single region of its helical structure. We found that the cytotoxic activity of mPep1 was about 9.6 fold-higher than that of Pep1 against HL-60 cells, as determined by the 3-(4,5-dimethylthiazol-2-yl)-5(3-carboxymethonyphenol)-2-(4 sulfophenyl)-2 H-tetrazolium (MTS) assay. In investigating the expression of phosphatidylserine, we observed that the native peptide (Pep1) caused both apoptotic cell death and necrotic cell death, depending on the concentration of the peptide. In contrast, the action of mPep1 was exclusively characteristic of necrotic cell death. This observation was further confirmed by agarose gel electrophoresis, in which clear ladder-like DNA bands were observed from cells exposed to Pep1, whereas DNA from cells treated with mPep1 produced a smeared pattern. We extended the study by investigating the release of mitochondrial cytochrome c into the cytosol, and the activation of caspase-3; both peptides caused the release of cytochrome c into the cytosol, and the activation of caspase-3.These results suggest that Pep1 may kill cancer cells by activating an apoptosis-inducing pathway, whereas mPep1 causes necrotic cell death by destroying cellular membrane structure notwithstanding sharing some cellular events with apoptotic cell death. PMID- 18425993 TI - An item-level analysis of the Center for Epidemiologic Studies Depression Scale (CES-D) by race and ethnicity in patients with coronary artery disease. AB - INTRODUCTION: The Center for Epidemiologic Studies Depression Scale (CES-D) is a measure of depressive symptoms that is commonly used in clinical studies. Studies employing this measure often describe variations in aggregate scores. However, few studies have examined variations in response to specific items by race and ethnicity. METHOD: This was a cross-sectional analysis among adults who were ages 50 and older and who were hospitalized for the management of coronary artery disease. Differential item functioning analysis was used to determine whether there were racial and ethnic variations in response to the items on the CES-D. RESULTS: Of the 571 participants, 76% were Caucasian, 12% were African-American, and 8% were Latino-American. After adjusting for age, gender, and education, four items exhibited differential item functioning. The odds of Latinos endorsing the item 'I was bothered by things that usually don't bother me' was 2.5 (95% CL 1.2 5.9); 2.7 (95% CL 1.4-5.1) for the item 'My sleep was restless'; 2.0 (95% CL 1.0 5.0) for the item 'I felt that I was just as good as other people'; and 2.3 (95% CL 1.3-4.5) for the item 'I enjoyed life'. The last two items were scored in reverse therefore; higher scores reflect more depressive symptoms. African Americans were 3.0 times more likely to endorse the item 'people were unfriendly' (95% CL 1.4-6.5). CONCLUSIONS: In this study, differences in somatic, positive affective and interpersonal symptoms were associated with the higher prevalence of depressive symptoms observed among racial and ethnic minorities when compared to Caucasians. PMID- 18425994 TI - Reduced-rank proportional hazards regression and simulation-based prediction for multi-state models. AB - In this paper we address two issues arising in multi-state models with covariates. The first issue deals with how to obtain parsimony in the modeling of the effect of covariates. The standard way of incorporating covariates in multi state models is by considering the transitions as separate building blocks, and modeling the effect of covariates for each transition separately, usually through a proportional hazards model for the transition hazard. This typically leads to a large number of regression coefficients to be estimated, and there is a real danger of over-fitting, especially when transitions with few events are present. We extend the reduced-rank ideas, proposed earlier in the context of competing risks, to multi-state models, in order to deal with this issue. The second issue addressed in this paper was motivated by the wish to obtain standard errors of the regression coefficients of the reduced-rank model. We propose a model-based resampling technique, based on repeatedly sampling trajectories through the multi state model. The same ideas are also used for the estimation of prediction probabilities in general multi-state models and associated standard errors. We use data from the European Group for Blood and Marrow Transplantation to illustrate our techniques. PMID- 18425995 TI - Quetiapine decreases alcohol consumption, craving, and psychiatric symptoms in dually diagnosed alcoholics. AB - AIMS: Patients with dual diagnosis are often excluded from clinical trials although more than half of all individuals with Bipolar Disorder have a substance abuse problem at some point in their lifetime, representing a high-risk clinical population. The purpose of this study was to investigate the safety and efficacy of quetiapine in the treatment of alcohol dependence comorbid with disorders characterized by high levels of mood and behavioral instability. METHODS: Twenty eight subjects, after a detoxification period, were orally treated with flexible doses of quetiapine for 16 weeks. At each assessment patients were evaluated through the Obsessive Compulsive Drinking Scale (OCDS), the Visual Analogue Scale (VAS) for craving, the Clinical Institute Withdrawal Assessment for Alcohol (CIWA Ar), the Brief Psychiatric Rating Scale (BPRS), the Hamilton Depression Rating Scale (HDRS), the Young Mania Rating Scale (YMRS), and the Clinical Global Impression (CGI) scale. RESULTS: Forty-three percent of patients remained totally alcohol free, 32% patients relapsed, with an average of 15.4 drinking days in the period of the study (112 days) and 25% dropped-out. Significant reductions from baseline to exit were observed in the OCDS, VAS, BPRS, HDRS, and number of drinking days per week. Changes in alcohol craving correlated with psychiatric symptoms as to BPRS and HDRS, with the highest level of correlation evidenced for the HDRS items of insomnia. DISCUSSION: In this open-label study, quetiapine decreased alcohol consumption, craving for alcohol, and psychiatric symptoms intensity, maintaining a good level of tolerance. A strength of this study is that the use of quetiapine was not adjunctive with other pharmacological and non pharmacological treatment. Double-blind placebo-controlled studies are required with a larger study population to confirm these data. In the meantime, for a select group of psychiatric patients, quetiapine may offer some advantages in preventing relapse. PMID- 18425996 TI - Identification of CYP2D6 null variants among long-stay, chronic psychiatric inpatients: is it strictly necessary? AB - We identified the null variants *3,*4,*5,*6,*7 and *8 of the CYP2D6 gene [encoding for cytochrome P450 (debrisoquine hydroxylase)] in a group of 84 chronic-stay psychiatric inpatients with severe schizophrenia or related disorders and receiving treatment with one or more CYP2D6 substrates for years. We also studied a group of 100 healthy controls of similar ethnic origin (Spanish Caucasians). Three patients were poor metabolizers (PMs) for antipsychotic drugs according to their CYP2D6 genotype (i.e. homozygous for the *4 allele) but they exhibited no adverse drug reaction over the years despite chronic treatment with CYP2D6 substrates. We suggest that CYP2D6 genetic screening is more useful in other type of psychiatric patients, particularly in younger ones starting treatment protocols. PMID- 18425997 TI - A fluorescence approach of the gamma radiation effects on gramicidin A inserted in liposomes. AB - The fluorescence of tryptophan residues of gramicidin A (gA), bound to phosphatidylcholine liposomes contains valuable information about local changes in the environment of the molecule induced by gamma radiation. With this work, we aim to demonstrate that the gamma radiation effect on the peptide involves the action of free radicals, derived from water radiolysis and the process of lipid peroxidation. Basically, the methodology consists of the analysis of UV and fluorescence emission spectra of the peptide liposome complexes under control conditions and upon gamma irradiation. Free radical production was impaired by the removal of molecular oxygen or the presence of ethanol in the liposome suspension. The intensity of the tryptophan fluorescence was recorded as a function of the gamma radiation dose in the range of 0-250 Gy and the data were fitted with a single decay exponential function containing an additional constant term (named residual fluorescence). The correlation between the decrease in tryptophan fluorescence emission (D(c) = 80 +/- 10 Gy) and increase in gamma radiation dose indicates the partial damage of the tryptophan side chains of gA. O(2) removal or ethanol addition partially reduced the decay of the tryptophan fluorescence emission involving an indirect action of gamma radiation via a water radiolysis mechanism. The residual fluorescence emission (A(0)) increases in O(2) free buffer (98 +/- 13) and in 10% ethanol-containing buffer (74 +/- 34) compared to control conditions (23 +/- 5). Varying the dose rate between 1-10 Gy/min at a constant dose of 50 Gy, an inverse dose-rate effect was observed. Thus, our study provides evidence for the lipid peroxidation effect on the tryptophan fluorescence. In conclusion, this article sustains the hypothesis of the connection between the lipid peroxidation and structural changes of membrane proteins induced by gamma radiation. PMID- 18425999 TI - Children's rights and wrongs. PMID- 18426000 TI - To the editor. Use of transvaginal ultrasonography and sonohysterography. PMID- 18426001 TI - Primary PCI in ST-segment elevation myocardial infarction. PMID- 18426002 TI - Primary PCI in ST-segment elevation myocardial infarction. PMID- 18426003 TI - Lung transplantation and survival in children with cystic fibrosis. PMID- 18426004 TI - Lung transplantation and survival in children with cystic fibrosis. PMID- 18426005 TI - Outcomes of care by hospitalists. PMID- 18426006 TI - Outcomes of care by hospitalists. PMID- 18426007 TI - Outcomes of care by hospitalists. PMID- 18426008 TI - Central venous catheterization--subclavian vein. PMID- 18426009 TI - Central venous catheterization--subclavian vein. PMID- 18426010 TI - Central venous catheterization--subclavian vein. PMID- 18426011 TI - A pivotal medical-device case. PMID- 18426012 TI - A pivotal medical-device case. PMID- 18426013 TI - [Epidemiology of non-infectious diseases; priority tasks of the new field of occupational medicine]. AB - The article describes the principles of epidemiological research management in occupational medicine and methodological approaches to them. The authors define relative risk of diseases and their conditionality by causative environmental factors. PMID- 18426014 TI - [Tuberculosis and malaria global prophylaxis in the light of decisions of the Big Eight]. AB - At the present time about two million people, one third of the Earth's population, are carriers of tuberculosis agent. Though tuberculosis is curable disease, it continues to take away lives of about 4400 persons; most of them are young and are in the most productive age. The most active incidence rate of tuberculosis occurs in the countries of Africa to the south of Sahara (29% of all cases of tuberculosis per head); half of new cases of tuberculosis fall on Asian countries: Bangladesh, China, India, Indonesia, Pakistan, Philippines. The governments of Big Eight maintain activities that have stabilized morbidity of tuberculosis on a world scale. Over 11 years (1995 - 2006) World Health Organization (WHO) implemented the DOTS strategy (Directly Observed Treatment with Short course of chemotherapy) in 183 countries and tested it on 26 millions patients with tuberculosis. Global data acquisition in 2005 found out morbidity of tuberculosis in 59% (the aim is 70%) and successful cure in 84% cases (the aim is 85 %). In 2006 WHO started realization of the Global Plan "Stop tuberculosis" (2006 - 2012). At the present time Global Fund use about 17% its resources to finance programs against tuberculosis. These funds help to reveal 5 millions extra cases of tuberculosis and cure 3 millions patients in the network of DOTS. PMID- 18426015 TI - [Teaching of physiology on the Medical Faculty of Moscow University in XVIII century]. PMID- 18426016 TI - [Disorders of calcium and magnesium metabolism: a diagnostic approach]. PMID- 18426017 TI - Neuroleptic malignant syndrome induced by quetiapine overdose. PMID- 18426018 TI - [No cases of reported virus transmission]. PMID- 18426019 TI - Do we need Medicare? PMID- 18426020 TI - Do we need Medicare? PMID- 18426021 TI - Do we need Medicare? PMID- 18426022 TI - Do we need Medicare? PMID- 18426023 TI - Baumol's disease. PMID- 18426024 TI - Prevention. Latin AIDS Awareness Day kicked off at Capitol. PMID- 18426026 TI - Housing. Tenancy restored for evicted HIV-positive renter. PMID- 18426025 TI - UNAIDS: billions still needed for global HIV intervention. PMID- 18426027 TI - Advocacy groups provide testing guidance for CDC funds. PMID- 18426028 TI - School district cleared of harassment, defamation claims. PMID- 18426029 TI - Exposure. HIV-positive sex offender's rape conviction affirmed. PMID- 18426031 TI - CDC awards grants to increase testing for affected groups. PMID- 18426030 TI - Prisons. Claim of misused pepper spray on ill inmate survives. PMID- 18426032 TI - Healthcare management. PMID- 18426033 TI - Abstracts of the 2008 AAHPERD National Convention, Fort Worth, Texas, USA. PMID- 18426034 TI - [Abstracts of the 92nd Congress of the Japanese Society of Legal Medicine, April 23-25, 2008, Nagasaki, Japan]. PMID- 18426035 TI - Abstracts of the Swiss Society of Sleep Research, Sleep Medicine and Chronobiology, March 5-6, 2008, Solothurn, Switzerland. PMID- 18426036 TI - Abstracts of the 22nd Congress of the German-speaking Society of Intraocular Lens Implantation and Refractive Surgery, 15-16 February 2008, Heidelberg, Germany. PMID- 18426037 TI - Medicare Advantage: options for standardizing benefits and information to improve consumer choice. AB - The Medicare Advantage (MA) program offers beneficiaries a choice of private health plans as alternatives to the traditional fee-for-service Medicare program. MA plans potentially provide additional value, but as plan choices have proliferated, consumers contemplating their options have had difficulty understanding how they differ. Through "standardization" more consistent types of information and a limited number of dimensions along which plans vary--MA plans could reduce complexity and improve beneficiaries' ability to make informed choices. Such standardization steps would offer more meaningful variation in the health coverage options available to beneficiaries, Medicare officials and their community partners would find it far easier to educate beneficiaries about their health plan choices, and beneficiaries would better understand what they were buying. Standardization might also strengthen the ability of the market-based Medicare Advantage program to incorporate beneficiary preferences. PMID- 18426038 TI - Abstracts of the 24th World Congress of Pathology and Laboratory Medicine, 20-24 August 2007, Petaling Jaya, Malaysia. PMID- 18426039 TI - 2nd European Consensus Meeting on Foam Sclerotherapy 2006, Tegernsee, Germany. AB - RATIONALE: The spread of Foam Sclerotherapy has resulted in the renaissance of sclerotherapy as a non-invasive treatment method for varicosis. The use of sclerosant foam for various forms of varicosis has now become established world wide as safe and effective. An expanded European expert committee meeting in April 2006, in Tegernsee was prompted by new findings and continuous further development of the method, but also because it had not been possible to consider all the relevant aspects of Foam Sclerotherapy in depth at the first meeting. OBJECTIVES: To revise and update the results of the 1st European Consensus Meeting and to include new, important topics in the development of Foam Sclerotherapy. To provide practical information for less experienced colleagues. METHODOLOGY: The 29 participants were sent a comprehensive questionnaire in advance covering all the relevant aspects of Foam Sclerotherapy. The organisers drew up various preliminary statements on the basis of the results. During the meeting itself the participants revised and/or approved and/or rejected these statements. An additional analysis of the questionnaire data also enabled preparation of an illustrative description of the broad spectrum of individual procedures used in this form of treatment. RESULTS: Foam Sclerotherapy has become an established treatment option for varicosis and has undoubtedly improved the management of varicose veins. European experts met Tegernsee for a second time to revise and expand their previous recommendations. In addition, individual working groups focused extensively on important issues. The current consensus, recommendations and descriptions of the individual aspects of the methods concern such issues as the indications for Foam Sclerotherapy, concentration and volume of the liquid sclerosants, relative and absolute contraindications, access and puncture options as well as clinical and ultrasound-guided recording of the treatment results. This final document reflects the experts' opinions on the principles of the effective and, above all, safe use of sclerosant foam for various indications and correct monitoring of the results of Foam Sclerotherapy. PMID- 18426041 TI - Abstracts of the International Society for Cellular Oncology (ISCO) Congress, March 5-8, 2008, Amsterdam, The Netherlands. PMID- 18426040 TI - Abstracts of the 32nd Scandinavian Congress of Rheumatology, 30 January-3 February 2008, Levi, Finland. PMID- 18426042 TI - Abstracts of the 79th Annual Scientific Meeting of the Aerospace Medical Association, May 11-15, 2008, Boston, Massachusetts, USA. PMID- 18426044 TI - [Abstracts of the 36th Annual Congress of the Netherlands Society of Psychiatry, 9-11 April 2008, Amsterdam, The Netherlands]. PMID- 18426043 TI - Abstracts of the 28th Annual Conference on Dialysis, March 2-4, 2008, Orlando, Florida, USA. PMID- 18426045 TI - HIV/AIDS and the veterinary practitioner -- making a difference. PMID- 18426046 TI - Clinical snapshot: large osseous sequestrum of the distal right humerus. PMID- 18426047 TI - Puppy and kitten socialization. PMID- 18426048 TI - Feline adverse vaccine reactions. PMID- 18426050 TI - Platinum products and services guide. PMID- 18426049 TI - [Abstracts of the 105th Annual Meeting of Internal Medicine, 2008]. PMID- 18426051 TI - [1/6 -- The nurse in geriatrics]. PMID- 18426052 TI - [The Charter of Rights and Liberties of the Aged Dependent Person. 5/7 Respect for religious convictions]. PMID- 18426053 TI - [The distribution of drugs]. PMID- 18426054 TI - [1/6 -- Daily care of a health mouth]. PMID- 18426055 TI - Age Discrimination in Employment Act; retiree health benefits. Final rule. AB - The Equal Employment Opportunity Commission is publishing this final rule so that employers may create, adopt, and maintain a wide range of retiree health plan designs, such as Medicare bridge plans and Medicare wrap-around plans, without violating the Age Discrimination in Employment Act of 1967 (ADEA). To address concerns that the ADEA may be construed to create an incentive for employers to eliminate or reduce retiree health benefits, EEOC is creating a narrow exemption from the prohibitions of the ADEA for the practice of coordinating employer sponsored retiree health benefits with eligibility for Medicare or a comparable State health benefits program. The rule does not otherwise affect an employer's ability to offer health or other employment benefits to retirees, consistent with the law. PMID- 18426056 TI - Medicaid program; elimination of reimbursement under Medicaid for school administration expenditures and costs related to transportation of school-age children between home and school. Final rule. AB - Under the Medicaid program, Federal payment is available for the costs of administrative activities "as found necessary by the Secretary for the proper and efficient administration of the State plan." This final rule eliminates Federal Medicaid payment for the costs of certain school-based administrative and transportation activities because the Secretary has found that these activities are not necessary for the proper and efficient administration of the Medicaid State plan and are not within the definition of the optional transportation benefit. Based on these determinations, under this final rule, Federal Medicaid payments will no longer be available for administrative activities performed by school employees or contractors, or anyone under the control of a public or private educational institution, and for transportation from home to school. In addition, this final rule responds to public comments received on the September 7, 2007 proposed rule. PMID- 18426057 TI - The effect of stroop interference on the categorical perception of color. AB - In two experiments, we examined the effects of Stroop interference on the categorical perception (CP; better cross-category than within-category discrimination) of color. Using a successive two-alternative forced choice recognition paradigm (deciding which of two stimuli was identical to a previously presented target), which combined to-be-remembered colors with congruent and incongruent Stroop words, we found that congruent color words facilitated CP, whereas incongruent color words reduced CP. However, this was the case only when Stroop interference was presented together with the target color, but not when Stroop stimuli were introduced at the test stage. This suggests that target name, but not test name generation, affects CP. Target name generation may be important for CP because it acts as a category prime, which, in turn, facilitates cross category discrimination. PMID- 18426058 TI - Processing the presence, placement, and properties of a distractor in spatial language tasks. AB - A common way to describe the location of an object is to spatially relate it to a nearby object. For such descriptions, the object being described is referred to as the located object; the object to which it is spatially related is referred to as the reference object. Typically, however, there are many nearby objects (distractors), resulting in the need for selection. We report three experiments that examine the extent to which a distractor in the display is processed during the selection of a reference object. Using acceptability ratings and production measures, we show that the presence and the placement ofa distractor have a significant impact on the assessment of the spatial relation between the located and reference objects; there is also evidence that the properties of the distractor are processed, but only under limited conditions. One implication is that the dimension that is most relevant to reference object selection is its spatial relation to the located object, rather than its salience with respect to other objects in the display. PMID- 18426059 TI - Prior knowledge enhances the category dimensionality effect. AB - A study of the combined influence of prior knowledge and stimulus dimensionality on category learning was conducted. Subjects learned category structures with the same number of necessary dimensions but with more or fewer additional, redundant dimensions and with either knowledge-related or knowledge-unrelated features. Minimal-learning models predict that all subjects, regardless of condition, either should learn the same number of dimensions or should respond more slowly to each dimension. Despite similar learning rates and response times, subjects learned more features in the high-dimensional than in the low-dimensional condition. Furthermore, prior knowledge interacted with dimensionality, increasing what was learned, especially in the high-dimensional case. A second experiment confirmed that the participants did, in fact, learn more features during the training phase, rather than simply inferring them at test. These effects can be explained by direct associations among features (representing prior knowledge), combined with feedback between features and the category label, as was shown by simulations of the knowledge resonance, or KRES, model of category learning. PMID- 18426060 TI - Passive tactile feedback facilitates mental rotation of handheld objects. AB - Mental rotation of objects improves when passive tactile information for the rotating object accompanies the imagined rotation (Wraga, Creem, & Proffitt, 2000). We examined this phenomenon further using a within-subjects paradigm involving handheld objects. In Experiment 1, participants imagined rotating an unseen object placed on their upturned palms. The participants were faster at mental rotation when the object was rotated on their palm than when the object remained stationary. Experiment 2 tested whether the performance advantage would endure when the participants received tactile information for only the start- and endpoints of the rotation event. This manipulation did not improve performance, relative to a stationary control. Experiment 3 revealed that ambiguous tactile information, continuous with the rotation event but independent of object shape, actually degraded performance, relative to a stationary control. In Experiment 4, we found that continuous tactile rotation discrepant from imagined object movement also hindered performance, as compared with continuous tactile information aligned with imagined object movement. The findings suggest a tight coupling between tactile information specifying continuous object rotation and the corresponding internal representation of the rotating object. PMID- 18426061 TI - Recognition and position information in working memory for visual textures. AB - In three experiments, we examined connections between item-recognition memory and memory for item-position information. With sequences of compound gratings as study and probe items, subjects made either item-position judgments (Experiments 1 and 2), by identifying the serial position of the study item that matched the probe, or recognition judgments (Experiment 3), by judging whether the probe had or had not been presented in the study series. Integrating a summed-similarity account of recognition into a signal detection framework shows that the variance of summed similarities on lure trials (probe not present in the study series) exceeds the variance on target trials (probe present in the study series). This prediction is borne out by the empirical zROC functions, all of which had slopes that were greater than 1. Additionally, about 25% of correct recognitions were accompanied by incorrect item position identification. Misidentifications of item position arose from two sources--structural similarity and positional similarity which combined in an approximately additive fashion. PMID- 18426062 TI - Repetition blindness in sentence contexts: not just an attribution? AB - Selective "blindness" to repeated words in rapid serial visual presentation (RSVP) occurs even when omitting these words compromises sentence syntax and meaning. The contributions of lexical and contextual factors to this repetition blindness (RB) phenomenon were evaluated using three tasks that combined RB and ambiguity resolution paradigms. During an RSVP sentence, a repeated word and matched but incongruous control were presented simultaneously, and participants were asked to report the entire sentence, including only the appropriate word. Substantial RB was evident in impaired report of repeated targets, whereas report of nonrepeated targets was enhanced when the distractor was a repeat. Experiment 2 confirmed these results with reduced reporting requirements, and Experiment 3 demonstrated the independence of repetition and sentence congruity effects. Results across all contexts support a lexical account of RB, which assumes that reactivation and identification of rapidly repeated words are impaired due to the refractory nature of lexical representations. PMID- 18426063 TI - The role of pragmatic principles in resolving attachment ambiguities: evidence from eye movements. AB - In an eyetracking experiment, participants read sentences that contained a prepositional phrase (PP) that could be attached to one of two preceding verbs. To understand the sentence, readers had to select which verb would serve as the host for the PP. In some of the sentences, the critical verbs and the PPs were part of a subordinate clause in which lexical and syntactic factors were expected to matter, but discourse factors favoring attachment of the material to positions where it would be relevant to the main assertion of the sentence were not expected to matter. In other sentences, the critical material was tested in main clause contexts in which the main assertion principle was predicted to apply. Hierarchical linear modeling showed that online attachment preferences were affected by clause type (main vs. subordinate). Specifically, the preference for a local verb over a distant verb was greater when the critical material appeared within a subordinate clause than when it appeared within a main clause. This pattern of results can be explained by the operation of the relativized relevance or main assertion principle, which in our sentences meant that participants favored attachment of the PP to the first verb over the second. PMID- 18426064 TI - Processing modifier-head agreement in reading: evidence for a delayed effect of agreement. AB - The present study examined whether type of inflectional case (semantic or grammatical) and phonological and morphological transparency affect the processing of Finnish modifier-head agreement in reading. Readers' eye movement patterns were registered. In Experiment 1, an agreeing modifier condition (agreement was transparent) was compared with a no-modifier condition, and in Experiment 2, similar constructions with opaque agreement were used. In both experiments, agreement was found to affect the processing of the target noun with some delay. In Experiment 3, unmarked and case-marked modifiers were used. The results again demonstrated a delayed agreement effect, ruling out the possibility that the agreement effects observed in Experiments 1 and 2 reflect a mere modifier-presence effect. We concluded that agreement exerts its effect at the level of syntactic integration but not at the level of lexical access. PMID- 18426065 TI - When you name the pizza you look at the coin and the bread: eye movements reveal semantic activation during word production. AB - Two eyetracking experiments tested for activation of category coordinate and perceptually related concepts when speakers prepare the name of an object. Speakers saw four visual objects in a 2 x 2 array and identified and named a target picture on the basis of either category (e.g., "What is the name of the musical instrument?") or visual-form (e.g., "What is the name of the circular object?") instructions. There were more fixations on visual-form competitors and category coordinate competitors than on unrelated objects during name preparation, but the increased overt attention did not affect naming latencies. The data demonstrate that eye movements are a sensitive measure of the overlap between the conceptual (including visual-form) information that is accessed in preparation for word production and the conceptual knowledge associated with visual objects. Furthermore, these results suggest that semantic activation of competitor concepts does not necessarily affect lexical selection, contrary to the predictions of lexical-selection-by-competition accounts (e.g., Levelt, Roelofs, & Meyer, 1999). PMID- 18426066 TI - Conversation, speech acts, and memory. AB - Speakers frequently have specific intentions that they want others to recognize (Grice, 1957). These specific intentions can be viewed as speech acts (Searle, 1969), and I argue that they play a role in long-term memory for conversation utterances. Five experiments were conducted to examine this idea. Participants in all experiments read scenarios ending with either a target utterance that performed a specific speech act (brag, beg, etc.) or a carefully matched control. Participants were more likely to falsely recall and recognize speech act verbs after having read the speech act version than after having read the control version, and the speech act verbs served as better recall cues for the speech act utterances than for the controls. Experiment 5 documented individual differences in the encoding of speech act verbs. The results suggest that people recognize and retain the actions that people perform with their utterances and that this is one of the organizing principles of conversation memory. PMID- 18426067 TI - Individual differences in syntactic ambiguity resolution: readers vary in their use of plausibility information. AB - Two experiments investigated the relation between individual differences in working memory capacity and differences in the efficiency of syntactic processing. In one experiment, readers comprehended sentences containing main verb/reduced-relative ambiguities that all resolved to the reduced-relative interpretation. High-span (but not low-span) readers processed sentences more slowly when the sentences were biased to the preferred, main-verb interpretation than when they were biased to the reduced-relative interpretation. Moreover, high span (but not low-span) readers used information about the plausibility of the different interpretations even though low-span readers appeared to possess the requisite knowledge. In Experiment 2, readers received intensive exposure to sentences with main-verb/reduced-relative ambiguities. Exposure enhanced low-span readers' use of plausibility information. Moreover, the effect of exposure generalized to sentences that were not included in the training materials. PMID- 18426068 TI - Verbalizing events: overshadowing or facilitation? AB - Verbal overshadowing refers to the surprising effect whereby additional verbal information about a visual stimulus hinders its subsequent recognition. In two experiments, we analyzed the validity of this effect for event recognition across various conditions of presentation and testing. Participants observed events that were either followed (Experiment 1) or preceded (Experiment 2) by a verbal description. Results showed that verbal overshadowing occurred when the verbal description was presented after the visual presentation, independent of the distractor type. However, when the verbal description preceded the event, recognition performance was seen to improve when distractor items incompatible with the verbal description were used. The findings were interpreted in terms of two interacting mental representations, which differ both in their level of abstraction and in their accessibility. PMID- 18426069 TI - Awareness in contextual cuing with extended and concurrent explicit tests. AB - The term contextual cuing refers to improved visual search performance with repeated exposure to a configuration of objects. Participants use predictive cues derived from learned associations between target locations and the spatial arrangement of the surrounding distractors in a configuration--to efficiently guide search behavior. Researchers have claimed that contextual cuing can occur implicitly. The present experiments examined two explicit measures--generation and recognition. In Experiment 1, we found that contextual cuing information was consciously retrievable when the number of trials used in a generation test was increased, and the results also suggested that the shorter tests that were used previously were not statistically powerful enough to detect a true awareness effect. In Experiment 2, concurrent implicit and explicit (generation and recognition) tests were employed. At a group level, learning did not precede awareness. Although contextual cuing was evident in participants who were selected post hoc as having no explicit awareness, and for specific configurations that did not support awareness, we argue that awareness may nevertheless be a necessary concomitant of contextual cuing. These results demonstrate that contextual cuing knowledge is accessible to awareness. PMID- 18426070 TI - Easy comes, easy goes? The link between learning and remembering and its exploitation in metacognition. AB - The cue-utilization view in metacognition assumes that judgments of learning (JOLs) are based on inferences from mnemonic cues deriving from the online processing of items during learning. This view calls for a specification of the underlying heuristics, their validity in predicting memory performance, and the extent to which they are utilized. This study examines one such heuristic: easily learned, easily remembered (ELER). We first show that ease of learning, as indexed by self-paced study time and by the number of trials to acquisition, is indeed a valid cue for recall success. We then demonstrate that this correlation between learning and rememberingunderlies metacognitive predictions about the future recallability of different items. The results are discussed in terms of the idea that metacognitive judgments incorporate knowledge about the internal ecology of cognitive processes, much as the perception of the external world embodies knowledge about the ecological structure of the environment. PMID- 18426071 TI - Metacognition and learning about primacy and recency effects in free recall: the utilization of intrinsic and extrinsic cues when making judgments of learning. AB - Although memory researchers know about primacy and recency effects, it is unclear whether students are aware of these effects and incorporate them when making judgments of learning (JOLs). The present research examined how participants use serial position information (extrinsic cues) when making JOLs after studying each item and showed that participants rely on the intrinsic qualities of the items and underestimate primacy and recency effects. However, when participants made JOLs prior to studying each item and engaged in multiple study-test sessions, their JOLs accurately reflected recall, as well as when serial position information was explicitly provided during the study phase. The findings are interpreted in a cue utilization framework and suggest that under certain conditions, participants can predict primacy and recency effects. PMID- 18426072 TI - The effects of tests on learning and forgetting. AB - In three experiments, we investigated whether memory tests enhance learning and reduce forgetting more than additional study opportunities do. Subjects learned obscure facts (Experiments 1 and 2) or Swahili-English word pairs (Experiment 3) by either completing a test with feedback (test/study) or receiving an additional study opportunity (study). Recall was tested after 5 min or 1, 2, 7, 14, or 42 days. We explored forgetting by means of an ANOVA and also by fitting a power function to the data. In all three experiments, testing enhanced overall recall more than restudying did. According to the power function, in two out of three experiments, testing also reduced forgetting more than restudying did, although this was not always the case according to the ANOVA. We discuss the implications of these results both for approaches to measuring forgetting and for the use of tests in promoting long-term retention. The stimuli used in these experiments may be found at www.psychonomic.org/archive. PMID- 18426073 TI - The reappearance hypothesis revisited: recurrent involuntary memories after traumatic events and in everyday life. AB - Recurrent involuntary memories are autobiographical memories that come to mind with no preceding retrieval attempt and that are subjectively experienced as being repetitive. Clinically, they are classified as a symptom of posttraumatic stress disorder. The present work is the first to systematically examine recurrent involuntary memories outside clinical settings. Study 1 examines recurrent involuntary memories among survivors of the tsunami catastrophe in Southeast Asia in 2004. Study 2 examines recurrent involuntary memories in a large general population. Study 3 examines whether the contents of recurrent involuntary memories recorded in a diary study are duplicates of, or differ from, one another. We show that recurrent involuntary memories are not limited to clinical populations or to emotionally negative experiences; that they typically do not come to mind in a fixed and unchangeable form; and that they show the same pattern regarding accessibility as do autobiographical memories in general. We argue that recurrent involuntary memories after traumas and in everyday life can be explained in terms of general and well-established mechanisms of autobiographical memory. PMID- 18426075 TI - Lipid peroxides in type 2 diabetic patients with neuropathy. AB - Diabetes and its metabolic changes in peripheral nerves contribute to cause a decrease of nitric oxide production and diminished nerve blood flow. Since lipid peroxides are thought to be formed by free radicals and may play an important role in the development of vascular disease, we have investigated the possible relationship between lipid peroxides (measured as thiobarbitouric acid reacting substances (TBARS) in diabetic patients with peripheral neuropathy. Seventy-seven patients with Type 2 diabetes (39 neuropathic and 38 non-neuropathic) and 38 control subjects were studied. The neuropathy study group had significantly lower levels of TBARS, 3.5micromol/l (2.2-5.6, 95% confidence limits) compared to controls 4.5microm/l (3.08-6.8), p < 0.001 and to diabetics without neuropathy 4.9micromol/l (3.09-8.05), p < 0.001. No differences were found in metabolic control between the two diabetic groups. In the neuropathy group there was a negative correlation between the score for nerve dysfunction with the TBARS levels (r = - 0.42, p < 0.01). In conclusion, in diabetic patients with neuropathy there are abnormalities of TBARS levels. PMID- 18426074 TI - Distinctive encoding reduces the Jacoby-Whitehouse illusion. AB - We investigated the influence of distinctive encoding on the Jacoby and Whitehouse (1989) illusion. Subjects studied visually presented words that were associated with either an auditory presentation of the same word (nondistinctive encoding) or a picture of the object (distinctive encoding). In both conditions, words were visually presented on the recognition test, and half were preceded by brief repetition primes. Priming test items increased hits and false alarms in the auditory condition, demonstrating the Jacoby-Whitehouse illusion. This illusion was reduced in the picture condition. In order to test whether this distinctiveness effect was caused by a recollection-based response strategy (i.e., the distinctiveness heuristic), we minimized recollection-based responding by having subjects make speeded recognition decisions. Contrary to the distinctiveness heuristic hypothesis, speeded responding did not eliminate the distinctiveness effect on the Jacoby-Whitehouse illusion. Picture encoding may reduce this illusion via a shift in preretrieval orientation, as opposed to a postretrieval editing process. PMID- 18426076 TI - Effects of experimental diabetes on hepatic drug metabolism in rats: the activities of flavin-containing monooxygenase, the phase II conjugation reactions and glutathione related enzymes. AB - Hepatic drug metabolism (flavin-containing monooxygenase (FMO), glutathione related enzymes, phase II conjugation reactions) and the hepatic contents of glutathione were investigated in normal rats, alloxan induced diabetic rats and streptozotocin (STZ) induced diabetic rats. The hepatic content of reduced or oxidized glutathione, the activities of glutathione related enzymes (glutathione reductase and glutathione peroxidase) and several enzymes (p-nitrophenol glucuronosyltransferase, aryl sulphotransferase I and II) involved in conjugation reactions were lower in alloxan- and STZ-induced diabetic rats than those in normal rats. In contrast, the activities of FMO and aryl sulphotransferase IV were significantly higher in alloxan- and STZ-induced diabetic rats than those in normal rats. Glutathione S-transferase (GST) activity also was remarkably higher in STZ-induced diabetic rats than that in normal rats. Insulin administered to STZ-induced diabetic rats prevented the hyperglycemia indicative of STZ-induced diabetes, but had no effect on the increased activities of GST. Another diabetogenic agent, alloxan, did not alter the activities of GST. On the other hand, the fluctuations in the enzymatic activities of FMO, UDP glucuronosyltransferase, aryl sulphotransferase and glutathione related enzymes were restored to normal level by treatment with insulin in both diabetic rats. These results show that STZ may be directly increasing activities of GST, and not as a result of the diabetic state the diabetogenic agent induces. However, the fluctuations of the activities of FMO, glutathione related enzymes and some phase II reactions were dependent on diabetic states. PMID- 18426077 TI - Diabetes evolution in rats after neonatal treatment with alloxan. AB - Physical exercises have been recommended in the prevention of non-insulin dependent diabetes mellitus (NIDDM), but the mechanisms involved in this intervention are not yet fully understood. Experimental models offer the opportunity for the study of this matter. The present study was designed to analyze the diabetes evolution in rats submitted to neonatal treatment with alloxan with the objective of verifying the suitability of the model to future studies with exercises. For this, newly born rats (6 days old) received intraperitoneal alloxan (A=200 mg/kg of body weight). Rats injected with vehicle (citrate buffer) were used as controls (C). The fasting blood glucose level (mg/dL) was higher in the alloxan group at the day 28 (C=47.25 +/- 5.08; A=54.51 +/- 7.03) but not at the 60 day of age (C=69.18 +/- 8.31; A=66.81 +/- 6.08). The alloxan group presented higher blood glucose level during glucose tolerance test (GTT) (mg/dL. 120 min) in relation to the control group both at day 28 (C=16908.9 +/- 1078.8; A=21737.7 +/- 1106.4) and at day 60 (C=11463.45 +/- 655.30; A=15282.21 +/- 1221.84). Insulinaemia during GTT (ng/mL. 120 min) was lower at day 28 (C=158.67 +/- 33.34; A=123.90 +/- 19.80), but presented no difference at day 60 (C=118.83 +/- 26.02; A=97.88 +/- 10.88). At day 60, the glycogen concentration in the soleus muscle (mg/100 mg) was lower in the alloxan group (0.3 +/- 0.13) in relation to the control group (0.5 +/- 0.07). No difference was observed between groups in relation to (micromol/g.h): Glucose Uptake (C=5.8 +/- 0.63; A=5.2 +/- 0.73); Glucose Oxidation (C=4.3 +/- 1.13; A=3.9 +/- 0.44); Glycogen Synthesis (C=0.8 +/- 0.18; A=0.7 +/- 0.18) and Lactate Production (C=3.8 +/- 0.8; A=3.8 +/- 0.7) by the isolated soleus muscle. The glucose-stimulated insulin secretion (16.7mM) by the isolated islets (ng/5 islets. h) of the alloxan group was lower (14.3 +/- 4.7) than the control group (32.0 +/- 7.9). Thus, we may conclude that this neonatal diabetes induction model gathers interesting characteristics and may be useful for further studies on the role of the exercise in the diabetes mellitus appearance. PMID- 18426078 TI - Effects of topical application of pirfenidone ointment on thermoplasty-induced acute lameness in a double-blind and acute and chronic lameness of musculoskeletal origin in an open multi-centered field trial in horses. AB - The effectiveness of pirfenidone ointment against thermoplasty-induced acute foreleg lameness in a double-blind study, and against acute and chronic lameness of musculoskeletal origin in an open multi-centered field trial was evaluated in this study. Thermoplasty was performed on both inner forelegs at designated locations of each horse under anesthetics. A 10% pirfenidone or placebo ointment was topically applied starting 24 hours after the thermoplasty three times daily for 7 days. For acute and chronic lameness of musculoskeletal origin, pirfenidone ointment was also applied one to three times daily for 7 to 10 days and continued for an additional 20 to 30 days. A marked swelling around the locations occurred in 24 hours post-thermoplasty. The topical application of pirfenidone ointment not only caused a significant reduction in the circumference measurements at 5, 6 and 7 days, but it also decreased the changes in the circumferences from pre thermoplasty as an index of edema, at 3, 4, 5, 6, and 7 days when compared to the placebo ointment at the corresponding times. Although treatment for 7 days of acute leg lameness of musculoskeletal origin with topical pirfenidone ointment caused significant decreases in swelling, heat, and pain, and improved the degree of flexion when compared with the pretreatment, it had little effect on chronic lameness except that it improved the flexion at the second-exam interval. It was concluded that topical application of pirfenidone is effective for treatment of acute lameness resulting from thermoplasty or from various types of musculoskeletal disorders, suggesting that pirfenidone offers a promising therapeutic potential to manage acute inflammation, an important component of lameness. PMID- 18426079 TI - Hematological malignancies are associated with a lower interferon-a blocking activity than solid tumors. AB - Interferon (IFN) and especially IFN-alpha exhibit clinical anti-tumor activity against various types of malignant diseases. Natural inhibitors to various cytokines and IFNs have been documented in vitro as well as in vivo. IFN inhibitors have been implicated for the ineffectiveness of IFN treatment in malignant neoplasias. The aim of this study was to investigate the incidence of the IFN inhibiting activity in serum from patients with haematological malignancies versus patients with solid tumours, as an effort to explain, just in part, the different response of these patients to IFN treatment. PATIENTS AND METHODS: Ninety patients with a clinically evident solid tumour and forty-six patients with haematological malignancies were included in the study. Serum samples from all patients were collected before any treatment and stored at -70 degrees until use. Controls sera were selected from 50 apparently healthy blood donors. Interferon-inhibiting activity as well as endogenous IFN-like activity were determined in all serum samples in a cell line highly sensitive to IFN. RESULTS: There was no endogenous IFN-like activity in any of the patients' group or controls' group. Sera from patients with haematological malignancies exhibited IFN-blocking activity at a lower percentage (21.7%) in comparison to sera from patients with solid tumours (56.6%, P<0.001), but at a significantly higher percentage in comparison to sera from controls (P<0.01). CONCLUSIONS: The fact that IFN inhibitors were detected at a significantly lower percentage in sera from patients with haematological malignancies versus patients with solid tumours, could explain in part the better response of the haematological malignancies to IFN treatment. PMID- 18426080 TI - The association of MMP-1, -3 and -9 genotypes with the prognosis of HCV-related hepatocellular carcinoma patients. AB - BACKGROUND/AIMS: Matrix metalloproteinases (MMPs) play important roles in cancer invasion and metastasis. Recently, functional gene polymorphisms in the promoter regions of MMP-1 1G/2G, MMP-3 5A/6A and MMP-9 C/T have been found, and have been reported to be associated with the prognosis of various cancers. In this study, we examined the association of the MMP-1, -3, -9 genotypes with the prognosis of hepatocellular carcinoma (HCC). METHODS: Ninety two patients with hepatitis C virus related HCC were enrolled in the study. Genomic DNA samples were extracted from peripheral white blood cells, and gene polymorphisms of MMP-1, MMP-3and MMP 9 were analyzed by PCR-RFLP methods. RESULTS: There were no significant differences of the survival rate between MMP-1 1G carriers and 2G homozygotes, and between MMP-9 T carriers and C homozygotes. By contrast, HCC patients with MMP-35A allele had a significantly poorer prognosis than 6A homozygotes. CONCLUSIONS: Unlike MMP-1 and MMP-9 genotypes, MMP-3 5A allele, with higher transcriptional activity, may be a risk factor for the poor prognosis of HCC patients. PMID- 18426081 TI - The HSP expression of passive repetitive plyometric trained skeletal muscle. AB - This study aims to understand the effect of ten-week passive repetitive plyometric (PRP) training on human skeletal muscle and the application of PRP training for performance. Vastus lateralis of nine candidates were aspirated before (pre) and after (post) PRP training. Histochemical approaches with regular hematoxylene-eosin (HE) and Mallory's phosphotungstic acid hematoxylin (PTAH) stains were used to demonstrate the changes of muscle fibers. Immunohistochemical studies with heat shock protein (anti-hsp72, Stressgen, Canada) were employed to display cellular activities. Each set of slides was quantitatively analyzed by using a modified morphometric method (Russ and Dehoff, 1999) on a Nikon ECLIPSE 80i microscope, equipped with an Evolution VF COOLED color video camera, and the Image-Pro Plus software (5.0 for Win; Media Cybernetics, USA). Finally, hsp72 mRNAs of both pre-PRP and post-PRP specimens were amplified through RT-PCR. Signal intensities were read by a densitometer and analyzed through the SPSS (11.0 for Win) statistically. Post-PRP muscle cells demonstrated hypertrophic change with increased cellular content and a narrowed inter-cellular space according to both HE and PTAH profiles. Post-PRP cellular hsp72 proteins were higher by up to five percent, as measured by a gray-scale reading. Further, after a training period of 10 weeks, hsp72 mRNA expression was several times higher. PMID- 18426082 TI - Displacement of opioid receptor binding ligands from the rat brain by N3-(2',5' dimethoxyphenacyl) arabinofuranosyluracil. AB - N3-(2',5 '-Dimethoxyphenacyl)arabinofuranosyluracil (N3-(2',5 '-DiMeOPhAc)AraU) is a pyrimidine nucleoside derivative which possesses antinociceptive effect by intracerebroventricular (i.c.v.) administration to mice. The compound (100 microM) significantly decreased the specific binding of [3H]D-Ala2, N-Me-Phe4, Gly5-ol-enkephalin (DAMGO) and [3H]D-Pen2, D-Pen5-enkephalin (DPDPE) at the mu- and delta-opioid receptor, respectively, but has no effect on the binding of [3H]U-69,593 at the kappa-opioid receptor of rat brain slices. The autoradiographic studies also demonstrated that [3H]DAMGO and [3H]DPDPE bindings on the rat brain slice were displaced by N3-(2',5'-DiMeOPhAc)AraU. These results indicate that N3-(2',5'-DiMeOPhAc)AraU interacts with mu- and delta-opioid receptors. PMID- 18426083 TI - Prostaglandin E1 transport into rat intestinal cryptic cell line IEC-6. AB - Administration of antitumor drugs such as 5-fluorouracil and methotrexate to rats causes intestinal injury. The intestinal injury was reported to be prevented by coadministration of prostaglandin E1 analogues to rats. IEC-6 cell is a useful in vitro model to study the cytoprotective effect in enterocytes, For its purpose, it is important to know whether or not prostaglandins are taken up into IEC-6 cells. The transport of prostaglandin E1 into IEC-6 cells was investigated, using [3H]-prostaglandin E1. Prostaglandin E1 was found to be transported into IEC-6 cells by the passive diffusion mechanism. PMID- 18426084 TI - Prevention of atherosclerosis in LDL receptor-mutant mice by human umbilical cord blood cells. AB - Previous studies have shown that congenic bone marrow transplantation into apolipoprotein E-deficient mice prevented hypercholesterolemia and atherosclerosis. In this study, we examined the effect of intravenous administration of human umbilical cord blood (HUCB) mononuclear cells on the progression of atherosclerosis in male homozygous mice that had mutation in the low-density lipoprotein receptor (Ldlr(tm1Her) mutation mice). In addition, the effect of human breastmilk alone as well as the combination of HUCB cells and breastmilk was studied on the prevention of atherosclerosis in these mice. In all groups of mice, atherosclerosis was predominant in the ascending aorta, but the rest of the aorta had variable evidence of atherosclerosis. Treatment of mice with HUCB cells significantly ameliorated the development of atherosclerosis in the ascending aorta, as compared with untreated mice; whereas breastmilk alone did not have any significant effect. A similar beneficial effect was observed with the combination therapy, which could be attributed only to HUCB cell treatment. There was no beneficial effect of treatment on the thoracic and abdominal aorta. Thus, early administration of HUCB cells prevents the progression of atherosclerosis in the ascending aorta of mice that are prone to the development of atherosclerosis. This beneficial effect occurred without any immunosuppression and graft-vs-host disease. PMID- 18426085 TI - The effect of isoniazid containing regimen on CYP2E1 during antituberculosis therapy. AB - Because isoniazid is a selective inducer of CYP2E1 and isoniazid-induced hepatotoxicity is believed to be due to activation of its metabolites by CYP450, this study was undertaken to determine the effect of isoniazid containing regimen on CYP2E1 in TB-patients. The activity of CYP2E1 in 11 newly diagnosed TB patients (5 F, 6 M) was investigated before (day 0) and during (day 14) treatment for tuberculosis. CYP2E1 activity was measured using the plasma metabolic ratio (MR) of 6-hydroxy-chlorzoxazone to chlorzoxazone, while CYP2E1 quantity in the peripheral lymphocytes was measured using SDS-PAGE. By day 14 of anti tuberculosis treatment, the activity of CYP2E1 was inhibited by 72% in 8 patients, but increased in 3 patients. The MR for the 8 patients was reduced from (Median & Range) 2.78 (1.1-21.5) on day 0, to 0.75 (0.4-1.22) on day 14, (P = 0.0006). Renal function was normal before and during the investigation. The detection of CYP2E1 by in peripheral lymphocytes was so variable that it could not be correlated with enzyme activity. Nevertheless, its detection in peripheral lymphocytes where normally is not resident indicates that CYP2E1 was induced by isoniazid. These results indicate that during treatment for tuberculosis with isoniazid containing regimen, CYP2E1 is induced but its activity is inhibited by isoniazid. PMID- 18426086 TI - Aberrant FHIT transcripts in human colorectal cancers. AB - The FHIT gene, located at human chromosome 3p14.2 spanning the FRA3B common fragile region, is frequently altered in several types of human cancers. To study the potential role of FHIT gene in colorectal cancer, expression of the FHIT gene were examined from 20 colorectal cancers for by reverse transcription-polymerase chain reaction (RT-PCR) and all of aberrant transcripts were cloned and sequenced. In addition, the effect of exogeneous rat FHIT overexpression on cell cycle was investigated by introducing the gene into normal rat kidney cells (NRK 52E). In RT-PCR, 7 cases of 25 patients with colorectal cancer showed 16 transcripts of abnormal sizes. Sequence analysis of the abnormal transcripts revealed these transcripts due to the deletion of multiple entire exons or part of exon sequences by errors in the splicing of pre-mRNA. The inserts of 59-bp and 138-bp sizes occurred in combination with in-frame deletions and was identified as part of the FHIT intron 5 sequence. In cell cycle analysis, over-expression of the FHIT gene in the FHIT-pTARGET-transformed NRK-52E cells did not affect cell proliferation and cell cycle distribution. Taken together, although alternative splicing of human FHIT is not directly associated with carcinogenicity, FHIT is frequently inactivated by exon skipping, intron retention, and activation of cryptic splice site within exon 6 in colorectal cancer. PMID- 18426087 TI - New aspects of cyclooxygenase-2 inhibition in myocardial infarction and ischaemia. AB - We have investigated the role of cyclooxygenase-2 (COX-2) in myocardial infarction (MI) and ischaemia in rabbits subjected to isoprenaline (ISP) a potent beta-adrenergic agonist. The acute phases of MI and repair mimicked those which occurred in humans. MI after induction with ISP was monitored by following increases seen in the levels of serum enzymes, Troponin I and Creatinine phosphokinase (CPK) in rabbits before and after ISP induced MI. Electrocardiographic (ECG) changes showed typical ST elevation and q-wave development. Histochemical studies of the rabbit heart on 2nd day after ISP injection showed changes of coagulation necrosis. Day 4 total coagulation with the loss of nuclear and striation associated with heavy interstitial infiltrate of neutrophils was found. Day 8 after infarction showed collagen deposition with capillary channels in between the remaining islands of myocytes in the infarcted area on the 16th day scarring was complete. Coronary perfusion rates (CPR) of the infarcted and nimesulide (a COX-2 inhibitor) treated rabbits displayed significant improvement on each corresponding day after infarction as compared to the infarcted and saline treated rabbits (P<0.01). These results suggest that nimesulide, a COX-2 inhibitor exhibit cardioprotective effects in MI. PMID- 18426089 TI - Competency of criminal defendants to waive assistance of counsel at trial. PMID- 18426088 TI - Effects of bacterial lipopolysaccharide on the pharmacokinetics of DA-8159, a new erectogenic, in rats. AB - Pharmacokinetic parameters of DA-8159 and one of its metabolites, DA-8164, were compared after intravenous and oral administration of DA-8159 at a dose of 30 mg/kg to control rats and rats pretreated with Klebsiella pneumoniae lipopolysaccharide (KPLPS). After intravenous and oral administration of DA-8159, most of the pharmacokinetic parameters of DA-8159 and DA-8164 were not significantly different between two groups of rats. This suggested that the pharmacokinetic parameters of DA-8159 and DA-8164 were not affected considerably by KPLPS. PMID- 18426090 TI - [The scientist-physician]. PMID- 18426091 TI - [Young football players aerobic perfomance in sub-maximum exercise with exhaustion at a moderate altitude without acclimation: experience in El Condor]. AB - ANTECEDENTS: With the same intensity of sub-maximum effort, the concentration of serum (LS) or muscular lactate is higher in Hypoxia conditions, or exercise at an altitude without acclimation, in relation with normoxia or at sea level (MSL). The highest level of lactate after fatigue or in a test of strict anaerobic condition, would not be modified when altitude changes without previous acclimation. OBJECTIVE: To abtain local data about the magnitude of the affect in SL of a effective altitude at 1,700 meters without acclimation, as a parameter of aerobic performance of young soccer players of our environment who live and train at sea level. VOLUNTEERS AND METHODS: Self-controlled experimental test with a number of 10 volunteers (16 - 18 years-old), soccer players of lower leagues from an important local sport club. Two tests in a cycle ergometer of a graded physical effort until fatigue, the first one in the Center of High Performance at MSL (474 meters) and the second one in the town El Condor (EC; 2.220 meters), province of Cordoba. Exercise Protocol: warm-up 2', 30 Km/h with 50W; 3consecutive steps of 3' + 50W each one; final phase 30 Km/h of 200W until fatigue and/or symptoms or maximum time of 15'. SL and HR measurement: basal; the last 30" of every step and in fatigue. SL determination with Accusport. Two tailed paired test. RESULTS: 1 volunteer dropped out befote completion in EC (n=9). Mean basal SL 2.1 mmol/L at MSL and EC (p=0.897). Mean difference (MD) EC vs. MSL +/- Standard error of 0.5 +/- 0.2 (p=0.033), 0.9 +/- 0.3 (p=0.008) and 0.6 +/- 0.3 mmol/L (p=0.103) in steps 1, 2, 3 respectively of sub-maximum effort. In fatigue (the highest level), Mean SL and Confidence Interval (CI) 95% of 7.6 (6.5-8.8) and 9.5 (7.8-11.2) mmol/L at MSL and EC respectively (p=0.030). Heart Rhythm (HR) without changes in EC in relation to MSL. 66% with a greater Borg scale and 33% of dizziness at EC. CONCLUSION: At an effective altitude at 1,700 meters without acclimation we can verify a lower aerobic performance with a subjective perception of a greater effort intensity, and neurological symptoms of hypoxia can be objectified in one-thirds of the cases. PMID- 18426092 TI - [Hight resoluton optical microscopy of the cellular cycle in psoriasis patients]. AB - Skin tissue from patients with Psoriasis was analyzed using HROM (High Resolution Optical Microscopy), studying epithelial differentiation and possible structural alterations of the queratinocytes. The samples were taken from 10 patients with histopathologic diagnosis of Psoriasis. This tissue samples where affixed with glutaraldehide buffer-collidine for 48 hours. Later processed with the HROM technique and colored with toluidine blue, metilene blue, basic Fuscine, and silver metenamine. The basal epithelial elements presented ovoid nucleus and most of them had prominent nucleolus. In 7 of the studied cases, the granulose stratus was absent, and thinner in the rest, with nucleus and nucleolus retention. At this level queratinocytes where observed with perinuclear anfofilia, as well as linfocitic and macrophagic infiltrate and union complex where elongated. PMID- 18426093 TI - [Liver resection in Privado Hospital's general surgery department: 13-year experience liver resections]. AB - BACKGROUND: Liver resection constitutes the treatment of choice in the liver primary neoplasms and some cases of metastatic tumors, but entail significant morbidity and mortality rates. OBJECTIVE: To present our experience in hepatic surgery. MATERIAL AND METHODS: We included 40 patients who underwent hepatectomies in our Institution from December 1991 through December 2004. The information was collected retrospectively and was analized descriptive and statistically. RESULTS: 40 patients underwent 40 hepatectomies. The diagnosis included primary benign hepatic disease (n= 9, 22.5%), primary hepatic malignancy (n= 7, 17.5%) and metastatic hepatic disease (n= 24, 60%). In the metastatic group, the most frecuent pathology was colonic adenocarcinoma (n= 17, 42%). There were 4 perioperatory deaths (10%) and the average hospitaly stay was 10.8 days. CONCLUSIONS: our data shows that the most frequent indication of hepatic resection in our experience was metastatic colonic adenocarcinoma as compared to the other Institutions with similar experience. PMID- 18426095 TI - [Salvage surgery treatment in osteosarcoma of the fibula with seventeen years survival]. AB - INTRODUCTION: Osteosarcoma is the most frequent primary bony tumor with two picks of incidence, one around the 15 years (adolescents) and another in old people. CLINICAL CASE: Patient of 14 years with pain clinic in proximal fibula. It was diagnosed by radiological studies and biopsy of located osteosarcoma with infiltration of cortical and periostium and extension toward fibula epiphysis. After preoperatory chemotherapy she was carried out tumoral resection with margins of 5 cm. following approaches of Enneking (surgery of salvation of the member). Several cycles of chemotherapy were completed after surgery. The patient, after seventeen years, is free of illness. DISCUSSION: Traditionally in the treatment of the located osteosarcoma you proceeded to the radical amputation for norm. At the moment due to the advances of the surgical and diagnostic techniques (able to detect precociously the local and distant dissemination of the tumor) it is possible to make other therapeutic modalities that preserve the member until in 90% of the patients. Thanks to the use of the chemotherapy the rate of survival nowadays approaches to 70% in those cases diagnosed previously to the metastasis appearance. At the moment in those cases in which the illness is located, the tumoral size is reduced and have a good answer to the preoperatroy chemotherapy, it is more advisable to opt for a surgery of salvation of the limb making a wise resection at distance of the margins of the tumor and accompanying it of cycles of combined chemotherapy. PMID- 18426094 TI - [Psycho-pathological risk factor of arterial disease (prevalence, superposition or vinculation with other factors)]. AB - Alexitimia and depression may or not coexist with others risk factors (comportment o physical). Frecuently they have relation with socio-echonomic status and with ethnia. Sometimes are determinants of the atherosclerotic process by increasing the vascular reactivity by the alteration of the evolution. There is no information in our country about this problem in general population. The present study result of the investigation of these aspects and the comportamental and physical factors of arterial disease, in a population of Cordoba province (Argentina Republic). PMID- 18426096 TI - [Leukaemia cutis: clinical manifestation of chronic lymphocytic leukaemia relapse]. AB - We present a 71 year old male patient with previous records of Chronic Lymphocytic Leukaemia who presented with a tumoral skin lesion. Histological and immunohistochemical studies confirmed the Leukaemia Cutis diagnosis. The patient underwent treatment with clorambucile and systemic steroids with remision of both haemathological and skin manifestation. The patient is still under close clinical follow up and remission continues eight months so far. PMID- 18426097 TI - [Evaluation in medicine by multiple choise examination]. PMID- 18426098 TI - Navigating orthodontic frontiers. PMID- 18426099 TI - Orthodontic treatment with preadjusted appliances and low-friction ligatures: experimental evidence and clinical observations. AB - AIM: To describe the features of low-friction ligatures during the leveling and aligning phase of fixed appliance therapy with preadjusted brackets. METHODS: Experimental in vitro and in vivo studies were carried out to test the performance of the low-friction system with regard to leveling and aligning of the dental arches, as well as to increasing the transverse dimension of the maxillary arch. RESULTS: The outcomes of experimental testing showed that the combination of the low-friction ligatures with the superelastic nickel-titanium wires produced a significantly smaller amount of binding at the bracket/archwire/ligature unit when compared to conventional elastomeric ligatures. CONCLUSION: The biomechanical consequences of the use of low-friction ligatures were shorter duration of orthodontic treatment during the leveling and aligning phase, concurrent dentoalveolar expansion of the dental arch, and the possibility of using biologically adequate orthodontic forces. PMID- 18426100 TI - Medicolegal perspective: interpretation of pretreatment orthodontic radiographs. AB - AIM: To detect the prevalence of anatomic anomalies and pathologic findings that could be screened from pretreatment orthodontic radiographs. METHODS: A thorough screening was conducted of the pretreatment orthodontic radiographs of a sample of 1,354 patients, collected by the author during the last 15 years from 4 locations, 3 in the Kingdom of Saudi Arabia and 1 in Egypt. RESULTS: A total of 154 significant anomalies and pathologic findings were recorded among the total sample group of 1,354 orthodontic patients (11.4%). Among these, 22 could have a serious effect on the general health of the patient. The findings included fractured odontoid process of the axis vertebra, os odontoideum, spondylolisthesis, fractured surgical needle in the oropharynx, fractured transpalatal archwire swallowed and stuck in patient's throat, unrecognized carotid artery stenosis, cystic lesions, and calcified stylohyoid ligaments. CONCLUSION: Medicolegal litigation is a main priority for the clinician, requiring cautiousness through screening of the pretreatment orthodontic radiographs beyond orthodontic purposes to exclude patients with significant anomalies and pathological findings. If not done from the medicolegal prospective, such screening should at least take place from an ethical perspective. The orthodontic clinician needs to know when to consult other medical specialties. PMID- 18426101 TI - Risk factors for post-orthognathic condylar resorption: a review. AB - AIMS: To evaluate the current evidence identifying risk factors for post orthognathic mandibular condylar resorption. METHODS: Studies published between January 1980 and August 2006 related to post-orthognathic condylar resorption were identified by searching the following databases: PubMed, Medline, EMBASE, PsycInfo, DARE, CENTRAL, and the Cochrane database of systematic reviews. The following keywords were used to identify relevant publications: condylar resorption, progressive condylar resorption, condylar atrophy, dysfunctional remodeling, and condylysis. A hand search of these papers was also carried out to identify additional articles. RESULTS: A number of methodological flaws are present within the current literature, including the comparison of nonmatched patient groups and poor imaging techniques, which makes evaluation difficult. Significant risk factors identified for condylar resorption include being female with mandibular retrognathia associated with an increased mandibular plane angle, the presence of pretreatment condylar atrophy, and undergoing posterior condylar displacement and upward and forward rotation of the mandible at the time of surgery. CONCLUSIONS: Better-controlled studies are required to fully understand the link between condylar resorption and orthognathic surgery. A number of risk factors have been identified within this article. It is important for orthodontists to consider these, particularly when consulting patients for treatment and identifying patients who may require closer postsurgical follow-up. PMID- 18426102 TI - Mandibular midline distraction osteogenesis: a complex case with severe crowding. AB - This article reports on a patient with severe maxillary and mandibular crowding treated by both expansion and extraction treatment. Two surgical procedures, surgically assisted palatal expansion and a mandibular midline osteotomy, were performed in sequence for skeletal expansion. A hybrid appliance, consisting of a Hyrax screw and 2 miniplates, was used in the mandible. Since expansion was not enough for relief of the crowding, 3 premolars were later extracted. The patient's final occlusion was satisfactory, but genioplasty may be desired in the future. PMID- 18426103 TI - Interdisciplinary management of a patient with skeletal anteroposterior discrepancy, impacted canine, and missing teeth. AB - For patients with complex dentofacial problems, orthodontic treatment is often not enough to obtain optimal esthetic and functional results. This case report demonstrates the treatment of a patient with skeletal anteroposterior discrepancy through an interdisciplinary approach and emphasizes the importance of collaboration with other dental disciplines to achieve the desired esthetic results. PMID- 18426104 TI - Orthodontic treatment of a patient with traumatic loss of maxillary incisors. AB - Obtaining and retaining a good treatment result for traumatically lost incisors can be difficult for the clinician. In this case report, the management of a patient with the chief complaint of missing maxillary central incisors due to trauma is presented. The patient had a constricted maxilla with a bilateral posterior crossbite and an end-to-end relationship anteriorly. Orthodontic intervention took 19 months. The cusps and palatal surfaces of the canines were ground and flattened to meet the patient's esthetic requests. PMID- 18426105 TI - Clinical experience with a new superelastic Ni-Ti-stainless steel retraction spring. AB - This article describes the clinical application of a new superelastic Ni-Ti stainless steel retraction spring used especially for canine retraction following the segmented archwire technique. The use of the new spring and its 2 configurations are demonstrated in a typodont experiment. Clinical experience with the new spring is presented in 2 patients; it produced canine retraction in these patients who required first premolar extraction. The use of the spring in both cases led to bodily tooth retraction without reactivation or any undesirable side effects. PMID- 18426106 TI - Maxillary second molar extractions in orthodontic treatment. AB - This article is a review of the rationales, indications, methods, and effects of orthodontic treatment with maxillary second molar extractions. In addition to the patient's malocclusion, specific considerations about the status and position of the maxillary second and third molars should be taken into account. In recent years, the development of temporary anchorage devices, in addition to extraoral traction and intraoral distalization appliances, has become another armamentarium in the distalization of the maxillary posterior teeth, which may affect the selection of teeth to be extracted from second to third molars. In conclusion, extraction of maxillary second molars is a viable option in selected cases at present, but it is important to understand the indications and limitations of this treatment choice. PMID- 18426107 TI - Miniscrews or palatal implants for skeletal anchorage in the maxilla: comparative aspects for decision making. AB - The advantages and disadvantages associated with placement, orthodontic treatment, and removal of miniscrews and palatal implants should be clearly understood by both the orthodontist and the patient. Device design, preoperative diagnostic measures, insertion and removal procedures, potential suprastructure application, implant migration and success rate, biomechanical anchorage facilities, and comparative risks and complications of both miniscrews and palatal implants must be considered if skeletal anchorage is intended to be used in the maxilla. This article reviews these and other factors to help clinicians decide which device should be selected for each individual case. PMID- 18426108 TI - VIP interview: Giuseppe Siciliani. PMID- 18426109 TI - A shocking waste. PMID- 18426111 TI - Land of plenty? AB - China will need to recruit around 150,000 nurses a year if ambitious plans to extend health care to all citizens are to be met. PMID- 18426110 TI - Nursing student attrition is costing taxpayers pounds 99 million a year. PMID- 18426112 TI - Third time healthy. AB - Since the founding of the People's Republic of China, health services have undergone major reforms. Now there are plans to move away from decentralisation. PMID- 18426113 TI - Supporting role. AB - Our nursing expertise series ends with a look at a nurse consultant in forensic mental health care. PMID- 18426114 TI - Heartbrealk and joy. PMID- 18426115 TI - Get back to work. PMID- 18426116 TI - Impact of national policy on the health of people seeking asylum. AB - Recent Department of Health policy has modified the stage in the application process that people seeking asylum are entitled to free NHS health care. This has caused confusion, not only among asylum seekers and settled refugees, but also among healthcare professionals. In turn, this has led to increased difficulty for people seeking asylum in accessing healthcare services. This article identifies when in the process asylum seekers are entitled to free NHS care. It considers how current legislation and the government stance on immigration are having a negative effect on the health of people seeking asylum while they are in the U.K., and to what extent nurses and other health professionals can help. PMID- 18426117 TI - Monitoring pulse rate. AB - This article, part of Nursing Standards clinical skills series, illustrates the sites for palpating the pulse and outlines the correct procedure for monitoring and recording pulse rate. This information will assist nurses to perform the skill safely as part of cardiovascular assessment of the patient. PMID- 18426118 TI - Delivering intravenous therapy in the community setting. AB - This article provides an overview of how an intravenous (IV) therapy service was developed in one primary care trust in England, the challenges that were faced and how they were overcome. The article includes some recommendations for others considering setting up a community IV therapy team. PMID- 18426119 TI - Infusion therapy part one: minimising the risks. AB - The role of the nurse in infusion therapy has evolved with advances i in vascular access technology and the variety of infusion devices available. With increasing numbers of patients requiring vascular access for a range of parenteral therapies, nursing staff need to demonstrate an understanding of the range and management of vascular access devices (VADs) and the clinical indications for individual devices. This article, the first of two, provides information on the type of VADs available and emphasises the importance o patient assessment to ensure safe, reliable and high-quality care for individual patients. The second part, to be published next week, examines some of the common complications associated with infusion therapy and identifies preventive measures. PMID- 18426120 TI - Post-operative pain. PMID- 18426121 TI - Knowledge in hand. PMID- 18426122 TI - Opportunity knocks. PMID- 18426123 TI - Safety airway management. PMID- 18426124 TI - My experiences within a specialist interest group. PMID- 18426126 TI - Spare us the gimmicks please! PMID- 18426127 TI - Tracheal intubation, tracheal tubes and laryngeal mask airways. AB - Tracheal intubation is used when a clear airway is difficult to achieve with a face mask or laryngeal mask, or if there is a risk of dislodgement of other forms of airway control. Tracheal intubation is considered to be an important part of patient management when a major intraoperative complication develops (for example anaphylaxis, massive haemorrhage or malignant hyperpyrexia). PMID- 18426128 TI - Awake intubation. AB - Securing the airway is a core skill in anaesthesia, the gold standard of which is tracheal intubation. Normally this is achieved after induction of anaesthesia. However, some circumstances demand an awake approach. Awake intubation can be achieved via several methods. Using the fibreoptic laryngoscope is the most widely used technique in the UK with minimal patient discomfort and a wide margin of safety. When compared with attempts at difficult direct laryngoscopy, awake fibreoptic intubation provides excellent cardiovascular stability when performed under good topical anaesthesia and conscious sedation. Understanding the equipment used as well as preparing the patient and being aware of potential pitfalls are important elements to performing a successful awake intubation. PMID- 18426129 TI - Basic concepts in mechanical ventilation. AB - Mechanical ventilatory support is a major component of the clinical management of critically ill patients admitted into intensive care. Closely linked with the developments within critical care medicine, the use of ventilatory support has been increasing since the polio epidemics in the 1950s (Lassen 1953). Initially used to provide controlled mandatory ventilation, today with advances in technology, most mechanical ventilators are triggered by the patient, increasing the awareness of the complexity of patient/ventilator interaction (Tobin 1994). Though ventilator appearance and design may have changed quite significantly and the variety of options for support extensive, the basic concepts of mechanical ventilatory support of the critically ill patient remains unchanged. This paper aims to outline these concepts so as to gain a better understanding of mechanical ventilatory support. PMID- 18426130 TI - Care of the intubated patient in the PACU: the 'ABCDE' approach. AB - This paper discusses airway management in the post anaesthetic care unit (PACU). Many patients will be extubated on arrival to the PACU, however a small number will need further support with tracheal intubation. Patient assessment is a key role for the PACU staff and using the ABCDE approach will provide a systematic method for assessing the patient and determining suitability for extubation. Care of the patient following extubation is also described. PMID- 18426131 TI - Theoretical and practical aspects of anaesthesia for thoracic surgery. AB - Thoracic surgical procedures account for only a small fraction of all surgery undertaken in the NHS. Thoracic surgery is performed in specialist centres as patients often suffer serious co-morbidities and require vigilant care and observation by staff involved in their treatment. Anaesthesia for thoracic surgery challenges the theoretical and practical experience of all involved. This review briefly summarises the anaesthetic skills and knowledge required to deliver a safe and professional service to patients with thoracic pathology. PMID- 18426132 TI - The first pneumonectomies for lung cancer. AB - The year 1933 was remarkable in the history of lung surgery in that five surgeons, in five separate medical centres in the USA and Canada, performed the first, and successful, pneumonectomies for tumours of the lung. PMID- 18426133 TI - On the cover. Hop. PMID- 18426134 TI - Differential diagnosis of chronic urticaria. AB - OBJECTIVES: To review diseases that can present with cutaneous signs and symptoms that mimic those observed in chronic urticaria and to discuss the workup necessary to distinguish these diseases from chronic urticaria. DATA SOURCES: We performed a PubMed search using the following keywords: urticaria, cryopyrin, Sweet syndrome, subacute cutaneous lupus, urticarial vasculitis, urticaria pigmentosa, angioedema, fixed drug eruption, bullous pemphigoid, and reticular erythematous mucinosis. Appropriate chapters in general dermatology textbooks were also reviewed. STUDY SELECTION: Articles that related to disease states, which present with persistent urticarial lesions, were catalogued for use in this review. RESULTS: Besides acute, chronic, and physical urticarias, there are 2 categories of diseases that have urticarial lesions. The first group includes those in which the skin lesions are almost indistinguishable from those seen in patients with chronic idiopathic urticaria. Thus, the diagnosis relies on a careful history and physical examination, and in some cases laboratory studies are required. The second group are ones that have skin lesions that at one point in their development have an urticaria-like appearance or on rare occasion may have such lesions. These latter diseases are numerous, and we have tried to highlight the ones that most mimic chronic idiopathic urticaria or are more common. CONCLUSIONS: A working knowledge of the diseases that can present with urticarial lesions is essential to accurately diagnose and effectively treat these symptomatic and sometimes serious conditions. PMID- 18426135 TI - Your patients are itching for answers. PMID- 18426136 TI - Ocular impact of intranasal corticosteroid therapy: all that surprising? PMID- 18426137 TI - Nasal ocular reflexes and eye symptoms in patients with allergic rhinitis. AB - BACKGROUND: Allergic patients often complain of eye symptoms during the allergy season. A possible mechanism for these eye symptoms is a nasal ocular reflex. OBJECTIVE: To demonstrate eye symptoms after nasal allergen challenge. METHODS: In a double-blind, placebo-controlled, crossover, clinical trial, 20 patients with seasonal allergic rhinitis were challenged in 1 nostril with antigen, and the response was monitored in both nostrils and in both eyes. Symptoms were recorded. Filter paper disks (intranasally) and Schirmer strips (intraocularly) were used for collecting secretions, which were subsequently eluted for the measurement of histamine and albumin levels. Patients were treated once topically at the site of challenge with azelastine or placebo. RESULTS: After placebo treatment, ipsilateral nasal challenge caused nasal symptoms and an increase in secretion weights; both were blocked by treatment with azelastine. Histamine and albumin levels increased only at the site of nasal challenge. Azelastine pretreatment inhibited the increase in albumin but not histamine levels. Symptoms of itchy and watery eyes increased significantly compared with symptoms with sham challenge after nasal allergen and were blocked by azelastine use. Ocular secretion weights increased bilaterally after placebo use and were not inhibited by azelastine use. CONCLUSIONS: Nasal allergen challenge releases histamine at the site of the challenge, which probably initiates a nasonasal and a nasal ocular reflex. This reflex is reduced by an H1-receptor antagonist applied at the site of the challenge. The eye symptoms associated with allergic rhinitis probably arise, in part, from a naso-ocular reflex. PMID- 18426138 TI - Prevalence and food avoidance behaviors for gustatory rhinitis. AB - BACKGROUND: Gustatory rhinitis is a type of nonallergic rhinitis that is usually associated with ingestion of hot or spicy foods. Characteristics of this condition and its impact on food choices have not been studied. OBJECTIVE: To survey individuals regarding causative foods, association with atopic conditions, and food avoidance behaviors for gustatory rhinitis. METHODS: An original, self administered questionnaire was distributed to children and adults who were seen in a busy outpatient dermatology clinic. Atopic history, frequency of rhinorrhea after food ingestion, intake of foods associated with gustatory rhinitis, symptom severity, and food avoidance were evaluated. RESULTS: Of the 571 participants who completed the survey, 396 (69%) indicated at least 1 food resulted in gustatory rhinitis symptoms. Patients with allergic rhinitis (P < .001) and a history of smoking (P = .049) were more likely to have experienced gustatory rhinitis. Bread (6%) and hot chili peppers (49%) represented the least and most common foods identified, respectively. A total of 65% of patients who experienced gustatory rhinitis never avoided the causative food, whereas 46% were never bothered by their symptoms. Patients who reported more causative foods were also more likely to use medications before eating to lessen their symptoms (P = .03). CONCLUSION: The results presented herein suggest that almost all foods can be implicated, both children and adults develop this condition, and patients with a history of allergic rhinitis or smoking are more likely to report gustatory rhinitis symptoms. Most patients though do not avoid the causative food(s) and are not significantly bothered by symptoms. PMID- 18426139 TI - Development of an inhaled endotoxin challenge protocol for characterizing evoked cell surface phenotype and genomic responses of airway cells in allergic individuals. AB - BACKGROUND: Environmental exposure to endotoxin is a known cause of exacerbation of asthma. Inhaled endotoxin protocols have been used to evaluate airway cell surface phenotypes associated with antigen presentation and innate immunity in healthy volunteers, but not in allergic volunteers. OBJECTIVES: To establish the safety of challenge with low-dose endotoxin (10,000 endotoxin units) (lipopolysaccharide [LPS]) inhalation in allergic individuals, to measure airway cell surface phenotypes associated with antigen presentation and innate immunity in induced sputum (IS) after LPS challenge, and to conduct gene expression profiling in IS cells to determine which host genetic networks are modified by LPS inhalation. METHODS: Induced sputum was obtained before and 6 hours after LPS inhalation in 10 allergic volunteers (8 with asthma and 2 with rhinitis). Flow cytometry was used to examine cell surface phenotypes on IS cells. Genomic expression was analyzed on a subset of IS samples (n = 10) using microarray and ingenuity pathway analysis. RESULTS: A total of 10,000 endotoxin units of LPS induced significant up-regulation of membrane CD14, CD11b, CD16, HLA-DR, CD86, and Fcepsilon receptor 1 on sputum phagocytes and increased expression of genes that influence antigen-presenting surface molecules (HLA-DR, chemokine ligand 2 or monocyte chemoattractant protein 1, v-rel reticuloendotheliosis viral oncogene homolog, prostaglandin-endoperoxide synthase 2 or cyclooxygenase 2, and transforming growth factor beta), immune activation (CD14, interleukin 1beta, and regulated upon activation, normal T cell expressed and secreted), and inflammation (intracellular adhesion molecule 1 and inhibitory kappaBalpha). Gene profiles for nuclear factor kappaB, interleukin 1, and tumor necrosis factor pathways were also significantly affected. CONCLUSIONS: Low-dose inhaled endotoxin challenge is safe in allergic individuals with mild to moderate disease. It enhances airway cell surface phenotypes and expression of genes associated with antigen presentation, innate immunity, and inflammation. Microarray with ingenuity pathway analysis can be successfully applied to sputum cells to characterize genetic responses to inhaled exacerbants. PMID- 18426140 TI - Physician adherence to the national asthma prescribing guidelines: evidence from national outpatient survey data in the United States. AB - BACKGROUND: The Expert Panel 2 Guidelines for the Diagnosis and Management of Asthma (EPR-2 guidelines) were developed to improve medication prescribing for patients with persistent asthma and to control acute exacerbations of asthma. In addition, these guidelines also encourage physician-provided asthma education. Little is known about prescribing adherence to EPR-2 guidelines. OBJECTIVES: To examine physician adherence to EPR-2 asthma medication prescribing guidelines and determine patient and physician factors associated with prescribing of asthma medications. METHODS: This study was a cross-sectional retrospective analysis of National Ambulatory Medical Care Survey physician visit survey data from 1998 through 2004. Data were extracted on all patients with an International Classification of Diseases, Ninth Revision (ICD-9) code for asthma (493.XX) and reason for visit as asthma. The unit of analysis was individual patient visit. The dependent variables in analyses were specific type of drug class. The independent variables were various patient and physician factors. Logistic regression analysis was used to evaluate study objectives. RESULTS: Asthma patients in 2002 had 3.3 times more odds of being prescribed controller medications compared with asthma patients in 1998. Findings in 2004 were not significant. Elderly patients had 54% less odds of receiving controller medication compared with those in the 35- to 64-year-old age group. Patients in the other race category are 40% as likely to receive controller asthma medication compared with white patients. Physicians in 2002 had 6.3 times more odds of prescribing long-acting beta-agonists compared with those in 1998. Physicians without ownership stake in their practice had 1.9 times more odds of providing asthma education to their patients compared with those who owned their practice. CONCLUSION: Physician prescribing of asthma pharmacotherapy does not adequately comply with EPR-2 treatment guidelines. PMID- 18426141 TI - Asthma-related medication use among children in the United States. AB - BACKGROUND: Asthma is one of the most common chronic conditions in children and has a major impact on health care use and quality of life. The Best Pharmaceuticals for Children Act mandates the federal government to sponsor pediatric studies of drugs approved for use in the United States but lacking evaluation in the pediatric population and lacking interest of commercial sponsors. As input into the drug selection and prioritization process, information is needed on the percentage of children who receive asthma-related medications. OBJECTIVE: To estimate the percentage of children who receive asthma related medications. METHODS: Retrospective analysis of outpatient medical and drug claims from members of commercial health care insurance plans enrolled any time from January 1, 2004, through December 31, 2005. The study population included 4,259,103 children throughout the United States aged birth through 17 years. RESULTS: Fifteen percent of all children were dispensed an asthma-related medication. Among 218,943 children with an asthma diagnosis, 188,286 (86%) had a dispensed asthma-related medication at any time during the 2-year study period. Among children without any asthma diagnoses, 398,880 (10%) had a dispensed medication. Fifty-nine percent of children with an asthma diagnosis were dispensed an anti-inflammatory medication within 90 days after a claim with a diagnosis of asthma. CONCLUSIONS: Asthma-related medications are dispensed to a large percentage of the pediatric population, including many who do not have claims with asthma diagnoses listed. Data on the pharmacokinetics and safety of these drugs in children are largely unknown and difficult to obtain. Clinical studies that use new tools and approaches are needed to resolve this information gap. PMID- 18426142 TI - Factors influencing dispensing of psychotropic medications to patients with asthma: a community pharmacy-based survey. AB - BACKGROUND: To our knowledge, the use of psychotropic medications (antidepressant, anxiolytic, and hypnotic agents) in patients with asthma has not been extensively explored. OBJECTIVE: To identify factors associated with the use of these medications in patients with asthma. METHODS: Regular customers (aged 18 50 years) of 348 pharmacies in 9 French regions were consecutively recruited from November 2003 to June 2004. Patients with a prescription for an asthma medication were included. Patients completed a questionnaire, complemented by the computerized records of their therapy dispensed in the past 12 months. Asthma control was measured with the Asthma Control Test. RESULTS: Among 886 patients (mean age, 37.4 years; 55.0% female), during the 12 months before the survey, the proportions who received at least 1 U of anxiolytic, antidepressant, and hypnotic agents were 25.6%, 13.7%, and 13.0%, respectively. Higher dispensing levels of antidepressant, anxiolytic, and hypnotic agents were observed with decreasing asthma control (P < .001 for each drug class). Other correlates of receiving any of these classes included older age, female sex, smoking status, and dispensing of antireflux treatment (P < .001 for all). These results were confirmed in multivariate analyses. CONCLUSIONS: Psychotropic drugs were commonly dispensed to patients with asthma, and correlates of receiving these drugs were identified in this population. The high use of psychotropic medication, particularly in patients with poorly controlled asthma, is of concern. PMID- 18426143 TI - The asthma emergency department visit: treating a crisis in the midst of uncontrolled disease. AB - BACKGROUND: Patients with asthma who require emergency department (ED) care are burdened with asthma symptoms, are at risk for hospitalization, and use expensive resources. OBJECTIVE: To examine whether an ED-based surveillance system that characterized asthma symptoms and care before, during, and after an ED visit enhances our understanding of the natural history of asthma exacerbations. METHODS: This cross-sectional follow-up enrolled 225 adult patients who presented to 1 of 6 Illinois EDs for asthma care. Clinical characteristics before ED presentation, care provided in the EDs, and 1-month follow-up status were assessed by self-administered questionnaire, medical record review, and telephone interview, respectively. RESULTS: Persistent asthma symptoms were reported by 85.8% and 84.9% (P = .37) of patients before their ED visit and follow-up call, respectively. For patients with persistent symptoms before the ED visit and follow-up call, 54.4% and 73.8% (P = .02) reported using an inhaled corticosteroid, respectively. Inhaled corticosteroids were recommended for 49.4% of discharged patients with persistent symptoms. Relapse rates for return ED visits and return hospitalizations were 26.4% and 9.6%, respectively. Patients had low asthma-specific and general quality-of-life scores at follow-up. CONCLUSIONS: Patients with asthma exacerbations most often had uncontrolled asthma before the ED visit that subsequently deteriorated, temporarily improved with ED treatment, and continued as uncontrolled asthma after the ED visit. Although improvements in care were reported 1 month after the ED visit, opportunities for additional improvement were observed. PMID- 18426145 TI - Survey of physicians' approach to food allergy, Part 2: Allergens, diagnosis, treatment, and prevention. AB - BACKGROUND: Food allergy (FA) practice varies widely. OBJECTIVE: To report differences between allergists and nonallergists regarding diagnosis, treatment, and prevention of FA. METHODS: A 2-page questionnaire was mailed to 3,000 allergists and 4,000 nonallergists. We previously published the findings on prevalence and manifestations. Herein, we report the findings on food allergens, diagnosis, treatment, and prevention. RESULTS: Responses were received from 584 (19.5%) of the allergists and 77 (1.9%) of the nonallergists. Because of the nonallergists' low response rate, descriptive comparisons were made without emphasis on statistical significance. Allergists and nonallergists differed in their rankings of the 5 most common food allergens. Nonallergists differed markedly from allergists in the diagnostic methods, using more leukocytotoxic tests (10.9% vs 0.3%), specific IgG4 tests (33.8% vs 6.0%), and intradermal tests (40.0% vs 9.5%), but fewer percutaneous skin tests (44.7% vs 98.9%), specific IgE tests (73.4% vs 97.8%), and challenges (61.1% vs 87.6%). They also differed in their use of open, single-blind, and double-blind challenge tests. Allergists were more likely to rely on elimination of proven food allergens and less likely to use conventional elimination diets, rotation diets, and sublingual or subcutaneous hyposensitization. Allergists were more likely to recommend a diet regimen during pregnancy (76.7% vs 35.3%) and lactation (91.1% vs 72.9%), breastfeeding (93.6% vs 84.3%), hydrolysate formulas (83.5% vs 64.3%), and withholding solids until the age of 6 months (89.4% vs 70.0%). CONCLUSION: Differences were noted between nonallergists and allergists regarding causes, diagnostic methods, treatment, and prevention of FA, indicating the need for more education in this area. PMID- 18426144 TI - Basophil histamine release activity and disease severity in chronic idiopathic urticaria. AB - BACKGROUND: Altered basophil degranulation phenotypes are found in patients with chronic idiopathic urticaria (CIU). OBJECTIVE: To evaluate CIU disease severity in relation to basophil histamine release (HR) characteristics. METHODS: Patients with CIU were recruited from allergy and dermatology clinics. Patients with recent use of systemic corticosteroids or immunosuppressants were excluded. Patients completed disease severity surveys and had blood basophils isolated and stimulated for HR using polyclonal goat anti-human IgE and N-formyl-met-leu-phe. The HR was measured using automated fluorometry. Multivariate linear regression analyses were used to investigate relationships between HR data and CIU disease measures. RESULTS: Fifty patients completed surveys, of which 34 were further categorized into 2 subgroups based on basophil HR response to anti-IgE stimulation: responders (> or = 10% HR) and nonresponders (< 10% HR). Responders and nonresponders reported similar use of oral corticosteroids, work absences, and quality-of-life impairment but differed in their patterns of medications used for CIU. Basophil responders had a trend of higher use of the emergency department for CIU management. Multivariate regression revealed that patients with the basophil responder phenotype experienced significantly higher current itch scores (P = .02) compared with nonresponders. CONCLUSIONS: Quality-of-life impairment is similar in CIU basophil subsets. Patients with CIU with a basophil responder phenotype report longer disease duration, a higher frequency of emergency department use, and significantly higher itch severity. PMID- 18426146 TI - Efficacy and safety of specific immunotherapy with a high-dose sublingual grass pollen preparation: a double-blind, placebo-controlled trial. AB - BACKGROUND: Sublingual immunotherapy (SLIT) is increasingly being used for the treatment of allergic rhinitis, but there are conflicting study results demonstrating clinically relevant efficacy. OBJECTIVE: To show clinical efficacy and safety of a new high-dose grass pollen preparation for SLIT. METHODS: In a 2 year, double-blind, placebo-controlled trial, 185 subjects with rhinitis or rhinoconjunctivitis, with or without asthma, were treated with a recently developed, high-dose, 6-grass pollen mixture for SLIT once daily. RESULTS: The primary end point, a combined symptom-medication score, showed almost no change in the placebo group during a 42-day evaluation period in the grass pollen season from 2003 to 2005, whereas active treatment was associated with a significant and clinically relevant improvement (full analysis set, P = .01; main data set, P = .002). The effect was irrespective of asthma diagnosis. Allergen-specific IgE showed no difference in both groups, and specific IgG4 and IgG1 increased with active treatment in the first and second study years compared with placebo, clearly indicating the immunogenic effect of the active treatment. The SLIT was well tolerated. No serious adverse drug reactions occurred. CONCLUSIONS: High dose, sublingual, specific immunotherapy with an extract of a 6-grass pollen mixture showed a significant and clinically relevant improvement in subjects with grass pollen-associated rhinitis or rhinoconjunctivitis, with or without asthma. The treatment with the sublingual solution was well tolerated. PMID- 18426147 TI - Clinical benefits of combination treatment with mometasone furoate nasal spray and loratadine vs monotherapy with mometasone furoate in the treatment of seasonal allergic rhinitis. AB - BACKGROUND: Intranasal corticosteroids and nonsedating antihistamines are the drug classes most often prescribed to treat allergic rhinitis (AR). Treatment guidelines recommend a combination of these agents for moderate-to-severe AR. However, clinical studies have found that combining an antihistamine with an intranasal corticosteroid provides few or no advantages over monotherapy with an intranasal corticosteroid. OBJECTIVE: To compare the efficacy of mometasone furoate nasal spray (NS) plus loratadine with that of monotherapy with the individual agents in patients 12 years and older with at least a 2-year history of seasonal AR. METHODS: In a multicenter, randomized, double-blind, parallel group, placebo-controlled clinical study, 702 patients were randomized to receive mometasone furoate NS, 200 microg, plus loratadine, 10 mg (n = 169); mometasone furoate NS, 200 microg (n = 176); loratadine, 10 mg (n = 181); or placebo (n = 176) once daily for 15 days. Primary efficacy variables were total nasal symptom score (TNSS) and total symptom score (TSS) as recorded on diary cards. RESULTS: No statistically significant differences were observed between mometasone furoate NS plus loratadine and mometasone furoate NS monotherapy for the primary efficacy variables. For TNSS and TSS, all 3 active drug therapies were more effective than placebo (P < or = .02). Both mometasone furoate NS treatment regimens were more effective than loratadine or placebo for TNSS (P < .01 for both) and TSS (P < or = .03 for both), whereas loratadine was more effective than placebo for TNSS only (P = .02). CONCLUSIONS: Combination therapy with mometasone furoate NS and loratadine provided benefits similar to monotherapy with mometasone furoate NS for the symptoms of seasonal AR. Therefore, mometasone furoate NS monotherapy was shown to be an effective treatment for seasonal AR. PMID- 18426148 TI - Clinical assessment and management of abnormal IgA levels. PMID- 18426149 TI - What is the proper concentration of histamine for use as a positive percutaneous skin test control? PMID- 18426150 TI - Myocardial dysfunction after epinephrine treatment of presumed insect venom allergy in a marathon runner. PMID- 18426151 TI - Down-regulation of atopic dermatitis-associated serum chemokines by wet-wrap treatment: a pilot study. PMID- 18426152 TI - Author's response to accuracy of IgE antibody laboratory results. PMID- 18426153 TI - Our broken health care system. PMID- 18426154 TI - Measuring clinical efficiency: issues in creating physician profiles. PMID- 18426155 TI - Work-related stress increases risk of CHD. PMID- 18426156 TI - Blood pressure control key in polycystic kidney disease. PMID- 18426157 TI - Identifying and managing nephrotic syndrome in adults. PMID- 18426158 TI - Use a stepped approach for atopic eczemia in children. PMID- 18426159 TI - Infantile paralysis. 1908. PMID- 18426160 TI - The perils of dental tourism, closer to home. PMID- 18426161 TI - The management of hypodontia: present and future. AB - Developmental absence of teeth (hypodontia) is associated with characteristic morphological changes in the teeth, alveolar volume deficiencies, and skeletal jaw mal-relationships. Management may be complex, involving several dental specialties, ideally working as a close-knit team. Improved diagnostic and treatment technologies continue to evolve, ever widening the management opportunities for these patients. This paper reviews current treatment methods and looks to what the future may hold. CLINICAL RELEVANCE: Specialist teams with access to traditional and evolving diagnostic and treatment technologies are best able to manage patients with complex treatment needs such as some of those presenting with hypodontia. PMID- 18426163 TI - Facial aesthetics: 1. Concepts and canons. AB - The clinical ability to alter dentofacial form requires an understanding of facial aesthetics. This is vital for any clinician involved in treatment that will alter a patient's dentofacial appearance, whether through orthodontics, facial growth modification, corrective jaw surgery or aesthetic dentistry. Part 1 of this article aims to cover the historical and theoretical aspects of facial aesthetics and their importance in contemporary dentofacial treatment. Part 2 will cover important aspects of the interview and clinical assessment of patients requiring alterations in their dentofacial appearance, including guidelines used in the assessment of facial proportions and symmetry. CLINICAL RELEVANCE: These articles cover the theoretical and clinical aspects of facial aesthetics required by clinicians involved in the treatment of dentofacial deformity. PMID- 18426162 TI - Periodontics: 9. Periodontitis and systemic conditions--is there a link? AB - The possibility that periodontal diseases have an effect on systemic health is being widely investigated. This paper reviews the potential links and the mechanism of these effects. CLINICAL RELEVANCE: Although the effects of periodontitis on systemic health do not yet have an effect on clinical decisions, knowledge and understanding of the link, effects and mechanism are important. PMID- 18426164 TI - Protaper--hybrid technique. AB - Crown down preparation is the most known and described technique since the introduction of Nickel Titanium (NiTi) rotary instruments in endodontics. This technique gives good results but has limitations, such as not addressing the initial anatomy of oval or dumb-bell shaped canals. The specific design of the Protaper instruments allows use of them with a different technique and, specifically, with a brushing motion in the body of the canal. The recent introduction of hand Protaper files has expanded the range of application of this system, especially in curved canals. The 'hybrid technique', using rotary and hand files, and the advantages of the combination of both instruments, are clearly described in this article. Used with this technique, the Protaper is a very safe system to use, and more controllable, for both inexperienced and experienced practitioners alike, than other systems. CLINICAL RELEVANCE: To understand the precautions needed with rotary files, and how to use them to preserve the anatomy of the canal and get a tapered shaping, even in severely curved canals. PMID- 18426165 TI - CAD/CAM restorations and secondary caries: a literature review with illustrations. AB - The literature related to failures of CAD/CAM restorations, with emphasis on secondary caries, is presented. A good approximal fit of the restoration is important to prevent plaque accumulation that may lead to secondary caries. CAD/CAM restorations allow inspection of the approximal fit prior to cementation and it may explain the relatively few failures due to secondary caries. The longevity of these restorations approaches that of gold restorations. CLINICAL RELEVANCE: The requirement for CAD/CAM restorations has advanced to a stage where high quality restorations with excellent anatomy and fit can be produced. The failure rate is relatively low compared to directly placed restorations. PMID- 18426166 TI - NICE guidelines and their relevance to the dental team. AB - The National Institute for Health and Clinical Excellence (NICE) of the National Health Service (NHS) produces guidance for the health sector on a range of issues. Specific guidance on dental subjects has been issued in relation to third molar removal, dental recall interval and HealOzone therapy. In addition, there are examples of more generic guidance which may also be relevant to dental practice. This paper discusses the background and functions of NICE and summarizes the guidance of relevance to the dental team. CLINICAL RELEVANCE: NICE guidance provides an authoritative summary of the current state of knowledge on certain clinical issues and gives guidance for healthcare workers. The authors discuss how this guidance should impact upon clinical practice. PMID- 18426167 TI - Tumoral calcinosis with dental manifestations--a case report. AB - Tumoral calcinosis (TC) is a rare entity causing abnormal periarticular calcifications in the affected joint observed in the first two decades of life. It is also known to affect the teeth. Few case reports on TC talk about the dental findings. This article presents the clinical, radiological and histological appearance of dental abnormalities in a 23-year-old female with TC. The differential diagnosis and dental management are discussed. CLINICAL RELEVANCE: Tumoral calcinosis should be considered in the differential diagnosis of abnormally short roots with complete or partial obliteration of pulp chambers in a young patient. PMID- 18426168 TI - Case 52. Physical signs for the general dental practitioner. Herpes simplex labialis. PMID- 18426169 TI - Aspects of human disease. PMID- 18426170 TI - Balance-billing with Blue Cross, and other miscellaneous questions. PMID- 18426171 TI - Morning huddle or prayer meeting? PMID- 18426172 TI - The art of "selling" dentistry. PMID- 18426173 TI - The dental benefits industry deals with change. PMID- 18426174 TI - High-tech dentistry: what is necessary and what is not. AB - Many new high-tech concepts and devices for dentistry have been introduced in recent years, and corresponding advertisements are predominant throughout the dental profession. Most high-tech devices and concepts are useful, some more than others. It would be difficult and costly to incorporate into dental practice every new technology that comes out on the market. It is common knowledge among most dentists that some dental procedures can be accomplished faster, easier and better with well-known, time-proven techniques, whereas other tasks are better accomplished with today's newest technologies. Dentists must weigh technology's perceived value to patients, as well as the relative efficiency of clinical techniques accomplished by conventional versus high-tech concepts when deciding whether to incorporate technology into their practices. This article has expressed my opinions about various high-tech concepts as I perceive them, and from my experience as I see them being accepted and used by the practicing profession. PMID- 18426175 TI - Your secret weapon: obnoxiously detailed narratives. PMID- 18426176 TI - The seven habits of highly effective dental teams. PMID- 18426177 TI - Experience success in every case presentation. PMID- 18426178 TI - Frontal sinus fractures: a review of trends, diagnosis, treatment, and outcomes at a level 1 trauma center in Connecticut. AB - The purpose of this retrospective study is to document the incidence and etiology of frontal sinus fracture at a level 1 traumacenterin Connecticut, assess the variability of management within our institution, and assess the effect on outcome over a seven-year period (January 1995-March 2002). Sixty-three cases were identified. Mean age was 31 years (92% male). The most common mechanism of injury was motor vehicle accident, followed by falls. The rates of associated facial fracture and intracranial injury were 83%, and 54% respectively. Diagnostic imaging missed two fractures. This study documents significant intrainstitutional variation in surgical management across contemporaneous primary treatment teams. There was no significant difference in outcome noted based on the treating specialty of the primary team for a given fracture type. The material used for sinus obliteration had no affect on outcome. Notable complications ofcerebrospinal fluid leak and mucocele are discussed as they contribute to current management guidelines. PMID- 18426179 TI - Salmonella typhimurium pulmonary infection in an immunocompetent patient. AB - A 55-year-old man presented to his primary care provider after a two-week history of worsening cough. He was admitted to the hospital and treated for community acquired pneumonia due to progression of symptoms and an abnormal chest radiograph. Chest computerized tomography demonstrated a large consolidation in the right upper lobe with areas of cavitation consistent with necrosis. Blood and sputum cultures were obtained, and the patient was subsequently diagnosed with pulmonary Salmonella typhimurium infection. The organism was isolated from a sputum specimen only. The patient had a history of chronic alcoholism, bronchitis, and esophageal dysmotility but no evidence of severe immunosuppression or malignancy. The patient responded well to antibiotic therapy with both symptomatic and radiologic improvement. As pulmonary Salmonella infection is exceedingly rare in the immunocompetent patient, a review of the literature is presented. PMID- 18426180 TI - Papillary fibroelastoma arising from the pulmonary valve associated with pulmonary embolization. AB - Cardiac papillary fibroelastomas are rare, usuallybenign tumors that can be detected at autopsy, during open-heart surgery, or with echocardiography. They usually arise from the cardiac valves and more commonly are found on the left side of the heart. Embolization of left-sided and tricuspid valve tumors has been well documented. This is the 1st reported case of pulmonary embolization of a papillary fibroelastoma arising from the pulmonary valve. PMID- 18426181 TI - Injuries in women's basketball. AB - Women's basketball has changed over time. It is a faster paced game than it was 30 years ago. Greatplayers, like Anne Meyers,who was the first, and only, woman to be signed to an NBA contract, would agree today's game is different. The game is played mostly "below the rim" but with players like Candice Parker, Sylvia Fowles and Maya Moore able to dunk the ball, the game is still changing. The one thing that remains constant in basketball, especially women's basketball, is injury. The majority of injuries in women's basketball are similar to those in men's basketball. Studies at the high school and college level show similar injury rates between women and men. ACL injuries are one exception, with female athletes having atwo to four times higher rate ofACL injuries. In this article, we review the common injuries in women's basketball. We discuss treatment issues and possible preventive measures. PMID- 18426182 TI - Federation of State Medical Boards' examination: an idea whose time has come. PMID- 18426183 TI - America's no. 1 public health problem. 1958. PMID- 18426184 TI - Report of the committee on scientific work. 1908. PMID- 18426185 TI - A pitch for disclosure. PMID- 18426186 TI - Traumatic brain injury: unnatural causes. PMID- 18426188 TI - Temporal and spatial organization of chemical and hydrodynamic processes. The system Pb(2+)-chlorite-thiourea. AB - Precise spatio-temporal organization of chemical, hydrodynamic, and mechanical processes is typical for biological systems where particular chemical reactions have to accrue in precisely assignment place and time. It is rarely studied and observed in chemical systems. We report unusual precipitation pattern formation of PbSO(4) in chemical media (Pb(2+)-Chlorite-Thiourea System). We have found that there is a region in a plane of initial concentrations of chlorite ions and thiourea where precipitation of lead sulfate appears in a form of ring if a pellet of lead nitrate is placed into the system. The whole process may be divided into three stages: movement of first circular front of lead containing solution, formation of a ringlike pattern of lead sulfate, and finally, propagation of this pattern resulting in a formation of ring with final inside diameter. Our experiments indicate that the following values are reproducible and quantify the PbSO(4) ring evolution: induction time, radius of the ring birth, speed of ring propagation toward the center, and final inside radius of the ring. Numerical solution of kinetic equations allowed us to give a qualitative explanation for the phenomenon observed. Formation and evolution of the PbSO(4) rings are caused by interplay of concentration gradients in the system and chemical reactions that occur in excitable chlorite-thiourea system. Chemical reactions and hydrodynamic processes form a complex causal network that made morphogenesis of this unusual pattern possible. PMID- 18426187 TI - Cancer-testis (CT) antigen expression in medulloblastoma. AB - Medulloblastoma is the most common childhood malignant tumor of the central nervous system. Treatment of medulloblastoma requires harmful therapy and nevertheless carries a poor prognosis. Due to their presence in various cancers and their limited expression in normal tissues, CT antigens are ideal vaccine targets for tumor immunotherapy. CT antigens, such as MAGE and NY-ESO-1, have been employed in clinical trials in various malignancies but little is known about their presence in medulloblastoma. We analyzed 25 medulloblastomas for the expression of a panel of CT antigens by RT-PCR and immunohistochemistry. Messenger RNA expression in the samples was as follows: GAGE 64%, MAGEA3/6 56%, SYCP1 44%, SLCO6A1 32%, MAGEC1 28%, MAGEC2 28%, MAGEA4 28%, NY-ESO-1 20%, MAGEA1 16%, and TPTE 0%. All cases except one (96%) were positive for mRNA expression of at least one CT gene. However, CT antigen expression was scarce on a protein level. Immunoreaction to monoclonal antibody E978 (NY-ESO-1) was negative in all cases; MA454 (MAGEA1), 57B (MAGEA4), M3H67 (MAGEA3/6), CT10#5 (MAGEC2) and #23 (GAGE) were each positive in 1 case, while the highest incidence of positive immunostaining, albeit heterogeneous, was seen with CT7-33 (MAGEC1) in 3 out of the 25 cases. The absence of correlation between mRNA and protein expression in medulloblastoma has not been observed in other tumors and further studies addressing the biology of CT antigens are necessary to investigate the present discrepant results. PMID- 18426189 TI - Computational study of the lowest triplet state of ruthenium polypyridyl complexes used in artificial photosynthesis. AB - The potential energy surfaces of the first excited triplet state of some ruthenium polypyridyl complexes were investigated by means of density functional theory. Focus was placed on the interaction between the geometrical changes accompanying the photoactivity of these complexes when used as antenna complexes in artificial photosynthesis and dye-sensitized solar cells and the accompanying changes in electronic structure. The loss process (3)MLCT --> (3)MC can be understood by means of ligand-field splitting, traced down to the coordination of the central ruthenium atom. PMID- 18426190 TI - Magnetic field effect on fluorescence in a mixture of N-ethylcarbazole and dimethyl terephthalate in a polymer film in the presence of electric fields. AB - Magnetic field effects on the fluorescence spectrum and on the electrofluorescence spectrum (plots of the electric field-induced change in fluorescence intensity as a function of wavelength) have been examined in electron donor and acceptor pairs of N-ethylcarbazole (ECZ) and dimethyl terephthalate (DMTP) in polymer films at different ratios of donor/acceptor concentration. In the mixture having a high concentration of ECZ, electric field induced quenching of the exciplex fluorescence originating from the photoinduced electron transfer becomes less efficient in the presence of a magnetic field. In the mixture having a low concentration of ECZ, on the other hand, no magnetic field effect was observed in the electrofluorescence spectrum, indicating that the hole carrier plays an important role in synergy effects of magnetic and electric field effects on exciplex fluorescence. In the absence of the applied electric field, the magnetic field does not affect either exciplex fluorescence with a peak at 450 nm or LE fluorescence emitted from the locally excited state of ECZ but enhances the broad emission with a peak at approximately 380 nm, probably assigned to the fluorescence of another type of exciplex between ECZ and DMTP. Thus, two kinds of magnetic field effects on fluorescence have been observed in a mixture of ECZ and DMTP in a polymer film. PMID- 18426191 TI - Rearrangement and hydrogen scrambling pathways of the toluene radical cation: a computational study. AB - A computational study is undertaken to provide a unified picture for various rearrangement reactions and hydrogen scrambling pathways of the toluene radical cation (1). The geometries are optimized with the BHandHLYP density functional, and the energies are computed with the ab initio CCSD(T) method, in conjunction with the 6-311+G(d,p) basis set. In particular, four channels have been located, which may account for hydrogen scrambling, as they are found to have overall barriers lower than the observed threshold for hydrogen dissociation. These are a stepwise norcaradiene walk involved in the Hoffman mechanism, a rearrangement of 1 to the methylenecyclohexadiene radical cation (5) by successive [1,2]-H shifts via isotoluene radical cations, a series of [1,2]-H shifts in the cycloheptatriene radical cation (4), and a concerted norcaradiene walk. In addition, we have also investigated other pathways such as the suggested Dewar Landman mechanism, which proceeds through 5, via two consecutive [1,2]-H shifts. This pathway is, however, found to be inactive as it involves too high reaction barriers. Moreover, a novel rearrangement pathway that connects 5 to the norcaradiene radical cation (3) has also been located in this work. PMID- 18426192 TI - Flat-structural motives in small alumino-carbon clusters C(n)Al(m) (n = 2-3, m = 2-8). AB - Small clusters consisting of a carbon diatom or triatom and several aluminum atoms are investigated ab initio, at an MP2 level of theory. The mainly ionic character of C-Al bonding predominantly leads to structures different from corresponding hydrocarbons (also if starting from analogous initial geometries), while still producing closed-shell ground states. It is found that in many cases stable geometries correspond to flat CAl(3) units. These include unique metal framed dicarbon and tricarbon all-flat species with unusual planar tetra coordination. Another frequent feature is a hyper-coordination of carbon atoms, supported by their high negative charges and critically examined via atom-in molecule calculations. Also characterized are anionic states, electronic excitation and ionization, electron attachment and detachment, and charge distributions. PMID- 18426193 TI - Excited-state proton transfer in methanol-doped ice in the presence of KF. AB - Steady-state and time-resolved emission techniques were employed to study the photoprotolytic cycle of an excited photoacid in ice in the presence of a low concentration of a weak base-like F(-). In previous studies we found that the photoprotolytic cycle in methanol-doped ice (1% mol fraction) is too slow to be observed at temperatures below 190 K. In this study we found that at temperatures below 240 K an additional proton-transfer process occurs in ice doped with 10 mM KF. We attributed this reaction to the creation of a mobile L-defect by F(-) ions. We used a diffusion-assisted reaction model, based on the Debye Smoluchowski equation, to account for the direct reaction of the L-defect with the excited photoacid at temperatures below T < 240 K. Below 160 K the spectroscopic properties as well as the photoprotolytic cycle change dramatically. We propose that below 160 K the sample enters a new phase. The excited-state proton-transfer (ESPT) process was observed and followed down to a liquid nitrogen temperature of approximately 78 K. In the low-temperature phase the ESPT rate is almost twice as much as at 180 K and the temperature dependence of the rate is very small. The kinetic isotope effect of the ESPT at the low temperature phase is small of about 1.3. PMID- 18426194 TI - Synthesis, characterization, and metabolic stability of porphyrin-peptide conjugates bearing bifunctional signaling sequences. AB - A series of four porphyrin-peptide conjugates bearing one linear bifunctional sequence containing a cell penetrating peptide (CPP) and a nuclear localization signal (NLS) were synthesized and their in vitro biological and stability properties investigated. All conjugates accumulated within human HEp2 cells to a significantly higher extent than their porphyrin-PEG precursor, and the extent of their uptake and cytotoxicity depends on the nature and sequence of the amino acids. Conjugates 2 and 5 bearing a NLS-CPP accumulated the most within cells and were the most phototoxic (IC50 approximately 7 microM at 1 J/cm2). All conjugates localized preferentially within the cell lysosomes, and in addition, conjugate 2 was also found in the ER. All conjugates were highly stable under nonenzymatic conditions, but their peptide sequences were cleaved to some extent (ca. 50% after 24 h) by proteolytic enzymes, such as cathepsin B, cathepsin D, prolidase, and plasmin. PMID- 18426195 TI - Combination of non-natural D-amino acid derivatives and allophenylnorstatine dimethylthioproline scaffold in HIV protease inhibitors have high efficacy in mutant HIV. AB - Several non-natural D-amino acid derivatives were introduced as P2/P3 residues in allophenylnorstatine-containing (Apns; (2S,3S)-3-amino-2-hydroxy-4-phenylbutyric acid) HIV protease inhibitors. The synthetic analogues exhibited potent inhibitory activity against HIV-1 protease enzyme and HIV-1 replication in MT-4 cells. Structure-activity relationships revealed that D-cysteine or serine derivatives contributed to highly potent anti-HIV activities. Interestingly, anti HIV activity of all the D-amino acid-introduced inhibitors was remarkably enhanced in their anti-HIV activities against a Nelfinavir-resistant clone, which has a D30N mutation in the protease, over that of the wild-type strain. HIV inhibitory activity of several analogues was moderately affected by an inclusion of alpha1-acid glycoprotein in the test medium. PMID- 18426196 TI - Discovery and optimization of triazolopyridazines as potent and selective inhibitors of the c-Met kinase. AB - Tumorigenesis is a multistep process in which oncogenes play a key role in tumor formation, growth, and maintenance. MET was discovered as an oncogene that is activated by its ligand, hepatocyte growth factor. Deregulated signaling in the c Met pathway has been observed in multiple tumor types. Herein we report the discovery of potent and selective triazolopyridazine small molecules that inhibit c-Met activity. PMID- 18426197 TI - Bootstrap-based consensus scoring method for protein-ligand docking. AB - To improve the performance of a single scoring function used in a protein-ligand docking program, we developed a bootstrap-based consensus scoring (BBCS) method, which is based on ensemble learning. BBCS combines multiple scorings, each of which has the same function form but different energy-parameter sets. These multiple energy-parameter sets are generated in two steps: (1) generation of training sets by a bootstrap method and (2) optimization of energy-parameter set by a Z-score approach, which is based on energy landscape theory as used in protein folding, against each training set. In this study, we applied BBCS to the FlexX scoring function. Using given 50 complexes, we generated 100 training sets and obtained 100 optimized energy-parameter sets. These parameter sets were tested against 48 complexes different from the training sets. BBCS was shown to be an improvement over single scoring when using a parameter set optimized by the same Z-score approach. Comparing BBCS with the original FlexX scoring function, we found that (1) the success rate of recognizing the crystal structure at the top relative to decoys increased from 33.3% to 52.1% and that (2) the rank of the crystal structure improved for 54.2% of the complexes and worsened for none. We also found that BBCS performed better than conventional consensus scoring (CS). PMID- 18426198 TI - Predictivity of QSAR. AB - A range of good quality, local QSARs for mutagenicity and carcinogenicity have been assessed and challenged for their predictivity in respect to real external test sets (i.e., chemicals never considered by the authors while developing their models). The QSARs for potency (applicable only to toxic chemicals) generated predictions 30-70% correct, whereas the QSARs for discriminating between active and inactive chemicals were 70-100% correct in their external predictions: thus the latter can be used with good reliability for applicative purposes. On the other hand internal, statistical validation methods, which are often assumed to be good diagnostics for predictivity, did not correlate well with the predictivity of the QSARs when challenged in external prediction tests. Nonlocal models for noncongeneric chemicals were considered as well, pointing to the critical role of an adequate definition of the applicability domain. PMID- 18426199 TI - Spectroscopic evidence for the formation of goldfingers. AB - Gold(I) has long been used in the treatment of rheumatoid arthritis, but the therapeutically relevant biological targets of gold(I) are not well understood. Here, we report the results of a spectroscopic investigation into the formation of goldfingers. By exploiting a thiolate to gold charge-transfer band in the UV, we observed that gold(I) interacts with zinc finger peptides with a stoichiometry of one gold ion for each two cysteine residues, forming 1:1, 1.5:1, and 2:1 adducts with zinc finger peptides containing CCHH, CCHC, and CCCC donor sets, respectively. In addition, circular dichroism experiments provided evidence that goldfingers are more ordered than the corresponding metal-free peptides but do not exhibit the canonical zinc finger structure. PMID- 18426200 TI - Gd7I12Zn: a group 12 atom in the octahedral Gd6 Cluster. AB - The title compound was synthesized from Gd, GdI 3 and Zn under Ar atmosphere at 850 degrees C. It crystallizes in the space group R3 (No. 148) with lattice constants a=15.686(1) A and c=10.4882(8) A. The structure features isolated Zn centered Gd 6 octahedra with all edges and corners capped by I atoms. The disorder of the Gd atoms is rationalized via electron microscopic techniques. A computational analysis using the extended Huckel method has been carried out in order to understand the bonding of this compound. The structure of isotypic La 7I12Co is also remarked (a=16.040(1) A and c=10.905(2) A). PMID- 18426201 TI - P123-assisted hydrothermal synthesis and characterization of rectangular parallelepiped and hexagonal prism single-crystalline mgo with three-dimensional wormholelike mesopores. AB - By adopting the strategy of dissolution-recrystallization under hydrothermal conditions (at 240 degrees C for 72 h) in the presence of a triblock copolymer (Pluronic P123), we fabricated nano- and microparticles of single-crystalline MgO of rectangular parallelepiped and hexagonal prism morphologies. The MgO crystallites display three-dimensional wormholelike mesopores and have a surface area as high as 298 m(2)/g even after calcination at 550 degrees C for 3 h. PMID- 18426202 TI - Synthesis, structures, and properties of group 9- and group 10-group 6 heterodinuclear nitrosyl complexes. AB - The reaction of the group 9 bis(hydrosulfido) complexes [Cp*M(SH)2(PMe3)] (M=Rh, Ir; Cp*=eta(5)-C 5Me5) with the group 6 nitrosyl complexes [Cp*M'Cl2(NO)] (M'=Mo, W) in the presence of NEt3 affords a series of bis(sulfido)-bridged early-late heterobimetallic (ELHB) complexes [Cp*M(PMe3)(mu-S)2M'(NO)Cp*] (2a, M=Rh, M'=Mo; 2b, M=Rh, M'=W; 3a, M=Ir, M'=Mo; 3b, M=Ir, M'=W). Similar reactions of the group 10 bis(hydrosulfido) complexes [M(SH)2(dppe)] (M=Pd, Pt; dppe=Ph 2P(CH2) 2PPh2), [Pt(SH)2(dppp)] (dppp=Ph2P(CH2) 3PPh2), and [M(SH)2(dpmb)] (dpmb=o C6H4(CH2PPh2)2) give the group 10-group 6 ELHB complexes [(dppe)M(mu S)2M'(NO)Cp*] (M=Pd, Pt; M'=Mo, W), [(dppp)Pt(mu-S)2M'(NO)Cp*] (6a, M'=Mo; 6b, M'=W), and [(dpmb)M(mu-S)2M'(NO)Cp*] (M=Pd, Pt; M'=Mo, W), respectively. Cyclic voltammetric measurements reveal that these ELHB complexes undergo reversible one electron oxidation at the group 6 metal center, which is consistent with isolation of the single-electron oxidation products [Cp*M(PMe3)(mu S)2M'(NO)Cp*][PF6] (M=Rh, Ir; M'=Mo, W). Upon treatment of 2b and 3b with ROTf (R=Me, Et; OTf=OSO 2CF 3), the O atom of the terminal nitrosyl ligand is readily alkylated to form the alkoxyimido complexes such as [Cp*Rh(PMe3)(mu S)2W(NOMe)Cp*][OTf]. In contrast, methylation of the Rh-, Ir-, and Pt-Mo complexes 2a, 3a, and 6a results in S-methylation, giving the methanethiolato complexes [Cp*M(PMe3)(mu-SMe)(mu-S)Mo(NO)Cp*][BPh 4] (M=Rh, Ir) and [(dppp)Pt(mu SMe)(mu-S)Mo(NO)Cp*][OTf], respectively. The Pt-W complex 6b undergoes either S- or O-methylation to form a mixture of [(dppp)Pt(mu-SMe)(mu-S)W(NO)Cp*][OTf] and [(dppp)Pt(mu-S) 2W(NOMe)Cp*][OTf]. These observations indicate that O-alkylation and one-electron oxidation of the dinuclear nitrosyl complexes are facilitated by a common effect, i.e., donation of electrons from the group 9 or 10 metal center, where the group 9 metals behave as the more effective electron donor. PMID- 18426203 TI - Solution [Cu(amm)]2+ is a strongly solvated square pyramid: a full account of the copper K-edge XAS spectrum within single-electron theory. AB - The solution structure of Cu(II) in 4 M aqueous ammonia, [Cu(amm)](2+), was assessed using copper K-edge extended X-ray absorption fine structure (EXAFS) and Minuit XANes (MXAN) analyses. Tested structures included trigonal planar, planar and D2d -tetragonal, regular and distorted square pyramids, trigonal bipyramids, and Jahn-Teller distorted octahedra. Each approach converged to the same axially elongated square pyramid, 4 x Cu-Neq=2.00+/-0.02 A and 1 x Cu-Nax=2.16+/-0.02 A (EXAFS) or 2.20+/-0.07 A (MXAN), with strongly localized solvation shells. In the MXAN model, four equatorial ammonias averaged 13 degrees below the Cu(II) xy plane, which was 0.45+/-0.1 A above the mean N4 plane. When the axial ligand equilibrium partial occupancies of about 0.65 ammonia and 0.35 water were included, EXAFS modeling found Cu-Lax distances of 2.16 and 2.31 A, respectively, reproducing the distances found in the crystal structures of [Cu(NH3)5](2+) and [Cu(NH3)4(H2O)](2+). A transverse axially localized solvent molecule was found at 2.8 A (EXAFS) or 3.1 A (MXAN). Six second-shell solvent molecules were also found at about 3.4+/-0.01 (EXAFS) or 3.8+/-0.2 A (MXAN). The structure of Cu(II) in 4 M pH 10 aqueous NH 3 may be notationally described as {[Cu(NH 3)4.62(H2O)0.38](solv)}(2+).6solv, solv=H2O, NH 3. The prominent shoulder and duplexed maximum of the rising K-edge XAS of [Cu(amm)](2+) primarily reflect the durable and well-organized solvation shells, not found around [Cu(H2O)5](2+), rather than two-electron shakedown transitions. Not accounting for solvent scattering thus may confound XAS-based estimates of metal-ligand covalency. [Cu(amm)](2+) continues the dissymmetry previously found for the solution structure of [Cu(H2O)5](2+), again contradicting the rack-bonding theory of blue copper proteins. PMID- 18426204 TI - Activation of N-sulfonyl oxaziridines using copper(II) catalysts: aminohydroxylations of styrenes and 1,3-dienes. AB - N-Sulfonyl oxaziridines are susceptible to electrophilic activation using copper(II) catalysts and react with styrenes under these conditions to provide 1,3-oxazolidines in a formal aminohydroxylation of the alkene. We propose a two step mechanism involving a cationic intermediate to account for the rate differences and regioselectivities observed using a variety of styrenes. In accord with our hypothesis, aminohydroxylations of a range of substrates bearing electron-stabilizing groups are successful, and 1,3-dienes are particularly good substrates for copper(II)-catalyzed aminohydroxylation. Reactions of unsymmetrical dienes provide good to excellent olefin selectivity, the sense and magnitude of which can be rationalized upon consideration of the stability of the cationic intermediates suggested by our mechanism. Diastereoselective synthesis of a diverse range of densely functionalized structures can be achieved by polyfunctionalization of dienes using aminohydroxylation as a key complexity increasing step. PMID- 18426205 TI - Experimental evidence for enzyme-enhanced coupled motion/quantum mechanical hydrogen tunneling by ketosteroid isomerase. AB - Although there are considerable data demonstrating that quantum mechanical hydrogen tunneling (HT) occurs in both enzymatic and nonenzymatic systems, little data exist that address the question of whether enzymes enhance the amount of HT relative to the corresponding nonenzymatic reactions. To investigate whether 3 oxo-Delta (5)-steroid isomerase (ketosteroid isomerase, KSI) enhances HT relative to the nonenzymatic (acetate-catalyzed) isomerization of Delta (5)-androstene 3,17-dione ( 1) to Delta (4)-androstene-3,17-dione ( 3), alpha-secondary deuterium kinetic isotope effects (KIE) at C-6 of the steroid were determined for both the KSI- and acetate-catalyzed isomerizations. The normal intrinsic secondary KIE for both wild type (WT) KSI (1.073 +/- 0.023) and acetate (1.031 +/ 0.010) suggest the possibility of coupled motion (CM)/HT in both the enzymatic and nonenzymatic systems. To assess the contribution of CM/HT in these reactions, the secondary KIE were also measured under conditions in which deuterium instead of hydrogen is transferred. The decrease in secondary KIE for WT (1.035 +/- 0.011) indicates the presence of CM/HT in the enzymatic reaction, whereas the acetate reaction shows no change in secondary KIE for deuterium transfer (1.030 +/- 0.009) and therefore no evidence for CM/HT. On the basis of these experiments, we propose that KSI enhances the CM/HT contribution to the rate acceleration over the solution reaction. Active site mutants of KSI (Y14F and D99A) yield secondary KIEs similar to that of WT, indicating that mutations at the hydrogen-bonding residues do not significantly decrease the contribution of CM/HT to the KSI reaction. PMID- 18426207 TI - Optical properties of ultrashort semiconducting single-walled carbon nanotube capsules down to sub-10 nm. AB - Single-walled carbon nanotubes (SWNTs) are typically long (greater than or approximately equal 100 nm) and have been well established as novel quasi one dimensional systems with interesting electrical, mechanical, and optical properties. Here, quasi zero-dimensional SWNTs with finite lengths down to the molecular scale (7.5 nm in average) were obtained by length separation using a density gradient ultracentrifugation method. Different sedimentation rates of nanotubes with different lengths in a density gradient were taken advantage of to sort SWNTs according to length. Optical experiments on the SWNT fractions revealed that the UV-vis-NIR absorption and photoluminescence peaks of the ultrashort SWNTs blue-shift up to approximately 30 meV compared to long nanotubes, owing to quantum confinement effects along the length of ultrashort SWNTs. These nanotube capsules essentially correspond to SWNT quantum dots. PMID- 18426208 TI - Marangoni flow-induced self-assembly of hexagonal and stripelike nanoparticle patterns. AB - We have developed a simple Marangoni flow-induced method for self-assembling nanoparticles (NPs) into both hexagonal and stripelike patterns. First, a NPs/ethanol suspension was spread on a slightly nonwettable and a wettable silicon oxide substrate. The Marangoni flow, induced by simultaneous evaporation of ethanol and condensation of water, leads to the formation of the corresponding hexagonal distributed circular NP rings and dotted stripes. The inter-ring spacing and ring size of the hexagonal patterns can be tuned by varying the relative humidity of the N2 stream blown over the slightly nonwettable substrate. Hexagonal patterns of circular NP patches can also be fabricated by lowering the evaporation of the condensed water droplets. On the wettable substrate, complex patterns result when the humidity of the N2 stream changes. PMID- 18426209 TI - Azafullerenes encapsulated within single-walled carbon nanotubes. AB - Methods of insertion of azafullerenes in single-walled carbon nanotubes (SWNTs) at different temperatures were investigated, while the effects of the conditions applied on the structure of azafullerene-based peapods, namely, C59N@SWNTs, were explored. Morphological characteristics of C59N@SWNTs were assessed and evaluated by means of high-resolution transmission electron microscopy (HR-TEM). Pathways and chemical reactions that occur upon encapsulation of C59N within SWNTs were evaluated. Monomeric azafullerenyl radical C59N. as inserted into SWNTs at high temperature, from purified (C59N)2 in the gas phase, can undergo a variety of different transformations forming dimers, oligomers or existing in its monomeric form inside SWNTs due to the stabilization effect by nanotube side walls. However, under milder conditions, that is, at lower temperature, bisazafullerene (C59N)2 can be inserted into SWNTs in its pristine dimeric form. PMID- 18426210 TI - New methods enabling efficient incorporation of unnatural amino acids in yeast. AB - New chemical and physical properties can be selectively introduced into proteins directly in live cells by genetically incorporating unnatural amino acids. The incorporation efficiency determines how effective such properties can be exploited and was very low in yeast. We developed a new method for efficient expression of orthogonal bacterial tRNA in yeast using polymerase III promoters that are cleaved from primary transcripts. In addition, a yeast strain deficient in nonsense-mediated mRNA decay was generated to prevent rapid degradation of target mRNA containing premature stop codons, which are the most frequently used to encode unnatural amino acids. These new strategies enabled a significant increase in yield of unnatural amino acid containing proteins from tens of micrograms to tens of milligrams per liter in yeast. PMID- 18426212 TI - Characterization and fate of black currant and bilberry flavonols in enzyme-aided processing. AB - The fate of black currant ( Ribes nigrum L.) and bilberry ( Vaccinium myrtillus L.) flavonols in enzyme-aided processing was studied. The flavonols were quantified and characterized by high-performance liquid chromatography equipped with a diode array detector and an electrospray ionization mass spectrometer. A tentative identification for 14 black currant and 19 bilberry flavonols is presented representing 11 previously unpublished conjugates. For the first time in any berry, the presence of laricitrin conjugates is reported. The enzyme-aided processing affected the flavonol extractability, elevating the yield in juices and decreasing that in press residues. Importantly, no significant loss of the berry flavonols was observed during the experiments, although some hydrolysis of flavonol conjugates was recorded. To maximize the effect on flavonol extractability, higher enzyme dosages were needed for black currants than for bilberries. The data show that the flavonol extractability and hydrolysis are dependent on the texture of raw material, the glycosylation pattern of the conjugates, and the activity profile of the enzyme preparation. PMID- 18426211 TI - Characterization of the two-component, FAD-dependent monooxygenase SgcC that requires carrier protein-tethered substrates for the biosynthesis of the enediyne antitumor antibiotic C-1027. AB - C-1027 is a potent antitumor antibiotic composed of an apoprotein (CagA) and a reactive enediyne chromophore. The chromophore has four distinct chemical moieties, including an ( S)-3-chloro-5-hydroxy-beta-tyrosine moiety, the biosynthesis of which from l-alpha-tyrosine requires five proteins: SgcC, SgcC1, SgcC2, SgcC3, and SgcC4; a sixth protein, SgcC5, catalyzes the incorporation of this beta-amino acid moiety into C-1027. Biochemical characterization of SgcC has now revealed that (i) SgcC is a two-component, flavin adenine dinucleotide (FAD) dependent monooxygenase, (ii) SgcC is only active with SgcC2 (peptidyl carrier protein)-tethered substrates, (iii) SgcC-catalyzed hydroxylation requires O 2 and FADH 2, the latter supplied by the C-1027 pathway-specific flavin reductase SgcE6 or Escherichia coli flavin reductase Fre, and (iv) SgcC efficiently catalyzes regioselective hydroxylation of 3-substituted beta-tyrosyl-S-SgcC2 analogues, including the chloro-, bromo-, iodo-, fluoro-, and methyl-substituted analogues, but does not accept 3-hydroxy-beta-tyrosyl-S-SgcC2 as a substrate. Together with the in vitro data for SgcC4, SgcC1, and SgcC3, the results establish that SgcC catalyzes the hydroxylation of ( S)-3-chloro-beta-tyrosyl-S-SgcC2 as the final step in the biosynthesis of the ( S)-3-chloro-5-hydroxy-beta-tyrosine moiety prior to incorporation into C-1027. SgcC now represents the first biochemically characterized two-component, FAD-dependent monooxygenase that acts on a carrier protein-tethered aromatic substrate. PMID- 18426213 TI - Influence of flavor solvent on flavor release and perception in sugar-free chewing gum. AB - The influence of flavor solvent [triacetin (TA), propylene glycol (PG), medium chained triglycerides (MCT), or no flavor solvent (NFS)] on the flavor release profile, the textural properties, and the sensory perception of a sugar-free chewing gum was investigated. Time course analysis of the exhaled breath and saliva during chewing gum mastication indicated that flavor solvent addition or type did not influence the aroma release profile; however, the sorbitol release rate was statistically lower for the TA formulated sample in comparison to those with PG, MCT, or NFS. Sensory time-intensity analysis also indicated that the TA formulated sample was statistically lower in perceived sweetness intensity, in comparison with the other chewing gum samples, and also had lower cinnamon-like aroma intensity, presumably due to an interaction between sweetness intensity on aroma perception. Measurement of the chewing gum macroscopic texture by compression analysis during consumption was not correlated to the unique flavor release properties of the TA-chewing gum. However, a relationship between gum base plasticity and retention of sugar alcohol during mastication was proposed to explain the different flavor properties of the TA sample. PMID- 18426214 TI - Mechanisms of flavor release in chewing gum: cinnamaldehyde. AB - Recently we reported that the release profile of cinnamaldehyde from a sugar-free chewing gum was correlated to the release of the sugar alcohol phase or was not in agreement with the log P model. The objective of this study was therefore to investigate mechanisms of cinnamaldehyde release from a sugar-free chewing gum; p cresol (similar log P value) was also analyzed for comparison. Breath analysis of the chewing gum samples over an 8 min consumption period reported that the maximum concentration of cinnamaldehyde was 2- to 3-fold higher during the initial phase of mastication in comparison to the later phase, whereas the concentration of p-cresol was relatively constant over these two time periods. By contrast the release profile of cinnamaldehyde from a flavored gum base (no sugar alcohol phase) was constant over the 8 min consumption period and similar to the release of cresol from the flavored gum base. On the basis of tandem mass spectrometry, cinnamaldehyde was reported to react with sorbitol and generate hemiacetal reaction products that were not stable under slight alkaline conditions; it was suggested to revert back to free cinnamaldehyde and sugar alcohol in the oral cavity. The increased polarity of these transient cinnamaldehyde-sorbitol hemiacetal reaction products would result in a more rapid release rate of cinnamaldehyde than would be typically predicted based on the affinity of cinnamaldehyde for the gum base. PMID- 18426215 TI - Fungal biotransformation of (+/-)-linalool. AB - The biotransformation of (+/-)-linalool was investigated by screening 19 fungi. Product accumulation was enhanced by substrate feeding and, for the first time, lilac aldehydes and lilac alcohols were identified as fungal biotransformation byproduct using SPME-GC-MS headspace analysis. Aspergillus niger DSM 821, Botrytis cinerea 5901/02, and B. cinerea 02/FBII/2.1 produced different isomers of lilac aldehyde and lilac alcohol from linalool via 8-hydroxylinalool as postulated intermediate. Linalool oxides and 8-hydroxylinalool were the major products of fungal (+/-)-linalool biotransformations. Furanoid trans-(2 R,5 R)- and cis-(2 S,5 R)-linalool oxide as well as pyranoid trans-(2 R,5 S)- and cis-(2 S, 5 S)-linalool oxide were identified as the main stereoisomers with (3 S,6 S) 6,7-epoxylinalool and (3 R,6 S)-6,7-epoxylinalool as postulated key intermediates of fungal (+/-)-linalool oxyfunctionalization, respectively. With a conversion yield close to 100% and a productivity of 120 mg/L.day linalool oxides, Corynespora cassiicola DSM 62485 was identified as a novel highly stereoselective linalool transforming biocatalyst showing the highest productivity reported so far. PMID- 18426216 TI - 3-Methylthiopropionic acid ethyl ester, isolated from Katsura-uri (Japanese pickling melon, Cucumis melo var. conomon), enhanced differentiation in human colon cancer cells. AB - The fully ripened fruit of Katsura-uri Japanese pickling melon ( Cucumis melo var. conomon) has rarely been used for food because the midripened fruit is utilized for making pickles, but the fully ripened fruit is no longer valuable for pickles due to the fruit body being too soft. We have considered the utilization of the fully ripened Katsura-uri fruit that may be used for nonpickling products, particularly if the fully ripened fruit demonstrated health benefits such as anticarcinogenic properties. The phytochemical extract from the fully ripened fruit of Katsura-uri Japanese pickling melon was purified via a bioassay-guided fractionation scheme, which was based on the induction of differentiation in a RCM-1 human colon cancer cell line. On the criteria of two differentiation markers (duct formation and alkaline phosphatase activity), the most potent fraction contained a compound identified as 3-methylthiopropionic acid ethyl ester, based on GC retention time, EI-MS, (1)H NMR, and (13)C NMR spectra. Previously, the role of 3-methylthiopropionic acid ethyl ester was considered as an odor producing compound in many fruits, but this study indicates potential medical benefits of this compound. PMID- 18426217 TI - Hydroxytyrosol prevents oxidative deterioration in foodstuffs rich in fish lipids. AB - Hydroxytyrosol, a natural phenolic compound obtained from olive oil byproduct, was characterized as an antioxidant in three different foodstuffs rich in fish lipids: (a) bulk cod liver oil (40% of omega-3 PUFAs), (b) cod liver oil-in-water emulsions (4% of omega-3 PUFAs), and (c) frozen minced horse mackerel ( Trachurus trachurus) muscle. Hydroxytyrosol was evaluated at different concentration levels (10, 50, and 100 ppm), and its antioxidant capacity was compared against that of a synthetic phenolic, propyl gallate. Results proved the efficiency of hydroxytyrosol to inhibit the formation of lipid oxidation products in all tested food systems, although two different optimal antioxidant concentrations were observed. In bulk oil and oil-in-water emulsions, a higher oxidative stability was achieved by increasing the concentration of hydroxytyrosol, whereas an intermediate concentration (50 ppm) showed more efficiency, delaying lipid oxidation in frozen minced fish muscle. The endogenous depletion of alpha tocopherol and omega-3 polyunsaturated fatty acids (omega-3 PUFAs) was also inhibited by supplementing hydroxytyrosol in minced muscle; however, the consumption of the endogenous total glutathione was not efficiently reduced by either hydroxytyrosol or propyl gallate. A concentration of 50 ppm of hydroxytyrosol was best to maintain a longer initial level of alpha-tocopherol (approximately 300 microg/g of fat), whereas both 50 and 100 ppm of hydroxytyrosol were able to preserve completely omega-3 PUFAs. Hydroxytyrosol and propyl gallate showed comparable antioxidant activities in emulsions and frozen fish muscle, and propyl gallate exhibited better antioxidant efficiency in bulk fish oil. PMID- 18426218 TI - Enantioselective synthesis of cyclic sulfamidates via Pd-catalyzed hydrogenation. AB - Using Pd(CF3CO2) 2/(S,S)-f-binaphane as the catalyst, an efficient enantioselective synthesis of cyclic sulfamidates was developed via asymmetric hydrogenation of the corresponding cyclic imines in 2,2,2-trifluoroethanol at room temperature with high enantioselectivities (up to 99% ee). PMID- 18426219 TI - Flexible protocol for the chemo- and regioselective building of pyrroles and pyrazoles by reactions of Danishefsky's dienes with 1,2-diaza-1,3-butadienes. AB - The versatility of the Mukaiyama-Michael-type addition/heterocyclization of Danishefsky's diene with 1,2-diaza-1,3-butadienes was applied to the synthesis of both 4 H-1-aminopyrroles and 4,5 H-pyrazoles. Thus, the same reagents furnished different types of highly functionalized azaheterocycles essentially depending on their structure: as a matter of fact, R1 = COOR or CONR 2 differently affects the acidity of the proton at the adjacent carbon. An unexpected formation of 5 H-1 aminopyrroles from the reactions carried out in water was also observed. PMID- 18426220 TI - Coupling of alkenes and alkynes: synthesis of the C1-C11 And C18-C28 fragments of miyakolide. AB - A transition metal-mediated, atom-economical approach toward the crucial A and D rings of miyakolide is described. A Pd-catalyzed alkyne-alkyne coupling/6- endo- dig cyclization is employed to assemble the A ring fragment. The key D ring pyran is constructed utilizing an Ru-catalyzed alkene-alkyne coupling followed by a Pd catalyzed allylic alkylation to establish the all-cis stereochemistry. PMID- 18426221 TI - Control of the photoreactivity of diarylethene derivatives by quaternarization of the pyridylethynyl group. AB - Photochromic behavior of diarylethene derivatives with (4-pyridyl)ethynyl group directly attached to the 6-pi hexatriene moieties of the diarylethenes was investigated. Upon quaternarization of the pyridine moieties, the photoreactivity was strongly suppressed. On the other hand, diarylethene derivatives with nonconjugated (4-pyridyl)ethyl group exhibited the photochromic reactivity, regardless of whether pyridyl rings are quaternarized or not. In the case of the (4-pyridyl)ethynyl-substituted compounds, the photochromic reactivity was suppressed by the addition of trifluoroacetic acid and was restored by diethylamine. PMID- 18426223 TI - Studies of sequence-specific DNA binding, DNA cleavage, and topoisomerase I inhibition by the dimeric chromomycin A3 complexed with Fe(II). AB - Chromomycin A3 (Chro) has been evidenced to exhibit much higher binding affinity toward Fe(II) by forming a highly stable 2:1 drug/metal complex, compared to its structural analogue, mithramycin (Mith). Different properties of the [(Chro)2 Fe(II)] complex acting on DNA, such as sequence specificity, DNA cleavage, and topoisomerase I (TopI) inhibition were studied. Kinetic analyses of surface plasmon resonance showed that the affinity of the [(Chro)2-Fe(II)] complex upon binding to hairpin DNA duplexes containing various tetranucleotide sequences follows the order: GGCC > CGCG > CCGG approximately GCGC > AGCT > ACGT > TGCA > TCGA. According to circular dichroism (CD) studies, most hairpin DNA duplexes appeared to retain their B-type conformations in the presence of the [(Chro)2 Fe(II)] complex, except the duplex containing the GGCC sequence, which exhibited the features of both A- and B-type DNA. In DNA-cleavage assays, the [(Chro) 2 Fe(II)] complex was shown to cause single-stranded cleavage of plasmid DNA because of a Fenton-type reaction. DNA cleavage activity of the [(Chro) 2-Fe(II)] complex was increased at low pH. Moreover, the complex was capable of inhibiting TopI activity. The [(Chro)2-Fe(II)] complex exhibited higher cytotoxicity than the [(Mith) 2-Fe(II)] complex in several cancer cell lines, most likely owing to its more stable dimeric structure and higher DNA-binding affinity. Our results provide significant evidence that the [(Chro)2-Fe(II)] complex could be promising in terms of its biological applications in the future. PMID- 18426222 TI - Structural basis for the inactivation of Thermus thermophilus proline dehydrogenase by N-propargylglycine. AB - The flavoenzyme proline dehydrogenase catalyzes the first step of proline catabolism, the oxidation of proline to pyrroline-5-carboxylate. Here we report the first crystal structure of an irreversibly inactivated proline dehydrogenase. The 1.9 A resolution structure of Thermus thermophilus proline dehydrogenase inactivated by the mechanism-based inhibitor N-propargylglycine shows that N5 of the flavin cofactor is covalently connected to the -amino group of Lys99 via a three-carbon linkage, consistent with the mass spectral analysis of the inactivated enzyme. The isoalloxazine ring has a butterfly angle of 25 degrees , which suggests that the flavin cofactor is reduced. Two mechanisms can account for these observations. In both, N-propargylglycine is oxidized to N propargyliminoglycine. In one mechanism, this alpha,beta-unsaturated iminium compound is attacked by the N5 atom of the now reduced flavin to produce a 1,4 addition product. Schiff base formation between Lys99 and the imine of the 1,4 addition product releases glycine and links the enzyme to the modified flavin. In the second mechanism, hydrolysis of N-propargyliminoglycine yields propynal and glycine. A 1,4-addition reaction with propynal coupled with Schiff base formation between Lys99 and the carbonyl group tethers the enzyme to the flavin via a three carbon chain. The presumed nonenzymatic hydrolysis of N-propargyliminoglycine and the subsequent rebinding of propynal to the enzyme make the latter mechanism less likely. PMID- 18426224 TI - Cross-bridge duty cycle in isometric contraction of skeletal myofibrils. AB - During interaction of actin with myosin, cross-bridges impart mechanical impulses to thin filaments resulting in rotations of actin monomers. Impulses are delivered on the average every tc seconds. A cross-bridge spends a fraction of this time (ts) strongly attached to actin, during which it generates force. The "duty cycle" (DC), defined as the fraction of the total cross-bridge cycle that myosin spends attached to actin in a force generating state (ts/ tc), is small for cross-bridges acting against zero load, like freely shortening muscle, and increases as the load rises. Here we report, for the first time, an attempt to measure DC of a single cross-bridge in muscle. A single actin molecule in a half sarcomere was labeled with fluorescent phalloidin. Its orientation was measured by monitoring intensity of the polarized TIRF images. Actin changed orientation when a cross-bridge bound to it. During isometric contraction, but not during rigor, actin orientation oscillated between two values, corresponding to the actin-bound and actin-free state of the cross-bridge. The average ts and tc were 3.4 and 6 s, respectively. These results suggest that, in isometrically working muscle, cross-bridges spend about half of the cycle time attached to actin. The fact that 1/ tc was much smaller than the ATPase rate suggests that the bulk of the energy of ATP hydrolysis is used for purposes other than performance of mechanical work. PMID- 18426225 TI - Cyclotides: natural, circular plant peptides that possess significant activity against gastrointestinal nematode parasites of sheep. AB - The cyclotides are a novel family of backbone-cyclized cystine-knot containing peptides from plants that have been shown to possess insecticidal activity against Helicoverpa larvae, an important pest of corn and cotton. In the current study, we investigated the in vitro effects of the cyclotides on the viability of egg, larval, and adult life stages of two species of economically important gastrointestinal nematode parasites of livestock, Hemonchus contortus and Trichostrongylus colubriformis. The cyclotides showed significant activity in inhibiting development of nematode larvae and motility of adult worms. Activities were comparable to some currently used anthelmintic compounds in these in vitro assay systems. A series of alanine mutants of the prototypic cyclotide kalata B1 were assayed against larvae to determine regions of the peptide responsible for activity. It was observed that anthelmintic activity was dramatically reduced as a consequence of the mutation of a large number of residues that are found clustered on one surface. Activities toward larvae were equivalent in the naturally occurring L-isomer of kalata B1 and a synthetic all-D-isomer, indicating that there is no chiral requirement for anthelmintic activity. The clustering of important residues and the lack of chiral selectivity further support the proposed mode of action of the cyclotides, which involves a membrane based interaction rather than an interaction at a specific receptor. The cyclotide-induced leakage of a fluorescent dye from vesicles used as a model membrane mimetic further confirms the membrane lytic ability of cyclotides. The relative potency of kalata B1 and kalata B2 in causing membrane leakage is consistent with the order of their anthelmintic activity. These results demonstrate that the cyclotides show potential for use in the control of gastrointestinal nematode parasites. PMID- 18426226 TI - Structural and mechanistic studies of arylalkylhydrazine inhibition of human monoamine oxidases A and B. AB - The structure and mechanism of human monoamine oxidase B (MAO B) inhibition by hydrazines are investigated and compared with data on human monoamine oxidase A (MAO A). The inhibition properties of phenylethylhydrazine, benzylhydrazine, and phenylhydrazine are compared for both enzymes. Benzylhydrazine is bound more tightly to MAO B than to MAO A, and phenylhydrazine is bound weakly by either enzyme. Phenylethylhydrazine stoichiometrically reduces the covalent FAD moieties of MAO A and of MAO B. Molecular oxygen is required for the inhibition reactions, and the level of O2 consumption for phenylethylhydrazine is 6-7-fold higher with either MAO A or MAO B than for the corresponding reactions with benzylhydrazine or phenylhydrazine. Mass spectral analysis of either inhibited enzyme shows the major product is a single covalent addition of the hydrazine arylalkyl group, although lower levels of dialkylated species are detected. Absorption and mass spectral data of the inhibited enzymes show that the FAD is the major site of alkylation. The three-dimensional (2.3 A) structures of phenylethylhydrazine- and benzylhydrazine-inhibited MAO B show that alkylation occurs at the N(5) position on the re face of the covalent flavin with loss of the hydrazyl nitrogens. A mechanistic scheme is proposed to account for these data, which involves enzyme catalyzed conversion of the hydrazine to the diazene. From literature data on the reactivities of diazenes, O2 then reacts with the bound diazene to form an alkyl radical, N2 and superoxide anion. The bound arylalkyl radical reacts with the N(5) of the flavin, while the dissociated diazene reacts nonspecifically with the enzyme through arylalkylradicals. PMID- 18426227 TI - Mapping local protein electrostatics by EPR of pH-sensitive thiol-specific nitroxide. AB - A first thiol-specific pH-sensitive nitroxide spin-label of the imidazolidine series, methanethiosulfonic acid S-(1-oxyl-2,2,3,5,5-pentamethylimidazolidin-4 ylmethyl) ester (IMTSL), has been synthesized and characterized. X-Band (9 GHz) and W-band (94 GHz) EPR spectral parameters of the new spin-label in its free form and covalently attached to an amino acid cysteine and a tripeptide glutathione were studied as a function of pH and solvent polarity. The pKa value of the protonatable tertiary amino group of the spin-label was found to be unaffected by other ionizable groups present in side chains of unstructured small peptides. The W-band EPR spectra were shown to allow for pKa determination from precise g-factor measurements. Is has been demonstrated that the high accuracy of pKa determination for pH-sensitive nitroxides could be achieved regardless of the frequency of measurements or the regime of spin exchange: fast at X-band and slow at W-band. IMTSL was found to react specifically with a model protein, iso-1 cytochrome c from the yeast Saccharomyces cerevisiae, giving EPR spectra very similar to those of the most commonly employed cysteine-specific label MTSL. CD data indicated no perturbations to the overall protein structure upon IMTSL labeling. It was found that for IMTSL, g iso correlates linearly with A iso, but the slopes are different for the neutral and charged forms of the nitroxide. This finding was attributed to the solvent effects on the spin density at the oxygen atom of the NO group and on the excitation energy of the oxygen lone-pair orbital. PMID- 18426229 TI - Supercritical fluid chromatography. PMID- 18426228 TI - On-line CE-LIF-MS technology for the direct characterization of N-linked glycans from therapeutic antibodies. AB - Glycan characterization of therapeutic proteins is of utmost importance due to the role of carbohydrates in protein stability, half-life, efficacy and mechanism of action. The primary assay for characterization and lot release of N-linked glycans on glycoprotein products at Genentech, Inc., is a capillary electrophoresis (CE) based assay, wherein PNGase F-released, APTS-labeled glycans are separated by CE with laser induced fluorescence (LIF) detection. With the growing number of new molecular entities in the pipeline, a fast and direct characterization approach is of increasing importance. This paper describes the development of CE-MS technology with on-line LIF detection that allows identification of major and minor glycan species (1-5% of total glycans) by providing accurate mass information. Data is presented for therapeutic rMAbs which presented previously unidentified, minor peaks during routine CE-LIF analysis. CE-LIF-MS was then used to provide accurate mass on these species, identifying CE peaks corresponding to sialylated (G1 + NANA, G2 + NANA), afucosylated (G0-GlcNAc-fucose) and low-level isomers of major APTS-labeled glycans G0, G1, G1' and G2. PMID- 18426230 TI - Controlled interparticle spacing for surface-modified gold nanoparticle aggregates. AB - Aggregation of gold nanoparticles of increasing size has been studied as a consequence of adsorption of 2-aminothiophenol (ATP) on gold nanoparticle surfaces. The capping property of ATP in the acidic pH range has been accounted from UV-vis absorption spectroscopy and surface-enhanced Raman scattering (SERS) studies. The effect of nanoparticle size (8-55 nm) on the nature of aggregation as well as the variation in the optical response due to variable degree of interparticle coupling effects among the gold particles have been critically examined. Various techniques such as transmission electron microscopy, X-ray diffraction, zeta-potential, and average particle size measurement were undertaken to characterize the nanoparticle aggregates. The aggregate size, interparticle distances, and absorption band wavelengths were found to be highly dependent on the pH of the medium and the concentration of the capping agent, ATP. The acquired SERS spectra of ATP relate the interparticle spacing. It has been observed that the SERS signal intensities are different for different sized gold nanoparticles. PMID- 18426231 TI - Colloidal stability of bare and polymer-grafted silica nanoparticles in ionic liquids. AB - The colloidal stability of bare and poly(methyl methacrylate) (PMMA)-grafted silica nanoparticles was studied in 1-alkyl-3-methylimidazolium ([C(n)mim])-based ionic liquids (ILs) with different anionic structures. The theoretical estimation of the colloidal interaction between monodispersed bare silica particles by using the Derjaguin-Landau-Verwey-Overbeek theory indicates that bare silica particles cannot be stabilized and they rapidly form aggregates in all the ILs used in this study. The instability of bare silica particles was experimentally confirmed by dynamic light scattering measurement and in situ transmission electron microscopy observations by utilizing the negligible vapor pressure of ILs. This evidence suggests that electrostatic stabilization is inefficient in ILs because of the high ionic atmosphere and the resulting surface-charge screening. The PMMA grafted silica particles exhibited long-term colloidal stability in [C(4)mim][PF(6)] and [C(n)mim][NTf(2)], which are compatible with the grafted PMMA. On the other hand, the PMMA-grafted particles could not be stabilized in [C 4mim][BF 4] due to the poor solubility of the grafted PMMA in the IL. Effective steric stabilization is important for obtaining stable colloidal particles in ILs. PMID- 18426232 TI - Fabrication of superhydrophobic cellulose-based materials through a solution immersion process. AB - An industrial waterproof reagent [(potassium methyl siliconate) (PMS)] was used for fabricating a superhydrophobic surface on a cellulose-based material (cotton fabric or paper) through a solution-immersion method. This method involves a hydrogen bond assembly and a polycondensation process. The silanol, which was formed by a reaction of PMS aqueous solution with CO 2, was assembled on the cellulose molecule surface via hydrogen bond interactions. The polymethylsilsesquioxane coatings were prepared by a polycondensation reaction of the hydroxyl between cellulose and silanol. The superhydrophobic cellulose materials were characterized by FTIR spectroscopy, thermogravimetry, and surface analysis (XPS, FESEM, AFM, and contact angle measurements). Analytical characterization revealed that nanoscale roughness protuberances uniformly covered the surface, thus transforming the cellulose from superhydrophilic to superhydrophobic with a water contact angle of 157 degrees . The superhydrophobic coatings were satisfactory with regard to both chemical and mechanical durability, and because of the transparency of the coatings the native cotton fabric displayed no changes with regard to either morphology or color. The easy availability of the materials and simplicity of this method render it convenient for mass production. PMID- 18426233 TI - Kinetics of solute adsorption at solid/aqueous interfaces: searching for the theoretical background of the modified pseudo-first-order kinetic equation. AB - It is shown that the modified pseudo-first-order (MPFO) kinetic equation proposed recently by Yang and Al-Duri simulates well the behavior of the kinetics governed by the rate of surface reaction and described by our general kinetic equation, based on the statistical rate theory. The linear representation with respect to time, offered by the MPFO equation seems to be a convenient tool for distinguishing between the surface reaction and the diffusional kinetics. Also, a method of distinguishing between the surface reaction and the intraparticle diffusion model based on analyzing the initial kinetic isotherms of sorption is proposed. The applicability of these procedures is demonstrated by the analysis of adsorption kinetics of the reactive yellow dye onto an activated carbon. PMID- 18426234 TI - Phase behavior for ternary systems composed of ionic liquid+saccharides+water. AB - In this work, we proposed a new aqueous biphasic system composed of hydrophilic ionic liquids (IL, 1-butyl-3-methylimidazolium tetrafluoroborate) and saccharides, forming an upper IL-rich phase and a lower sugars-rich phase. It was found that that the distance between binodal curves and the origin is the increasing order of sucrose 200). Two types of optically active binaphthol derivatives, 1-(2-hydroxy-6-(naphthalen-1-yl)naphthalen-1-yl)-6-(naphthalen-1 yl)naphthalen-2-ol and 6-butyl-1-(6-butyl-2-hydroxynaphthalen-1-yl)naphthalen-2 ol, were prepared by this chemo-enzymatic reaction protocol and were used as chiral templates for symmetric reactions. PMID- 18426239 TI - Solid-phase combinatorial synthesis of a lysyl-tRNA synthetase (LysRS) inhibitory library. AB - The solid-phase combinatorial synthesis of a new library with potential inhibitory activity against the cytoplasmic lysyl-tRNA synthetase (LysRS) isoform of Trypanosoma brucei is described. The library has been specifically designed to mimic the lysyl adenylate complex. The design was carried out by dividing the complex into four modular parts. Proline derivatives (cis-gamma-amino-L-proline or trans-gamma-hydroxy-L-proline) were chosen as central scaffolds. After primary screening, three compounds of the library caused in vitro inhibition of the tRNA aminoacylation reaction in the low micromolar range. PMID- 18426240 TI - Molecular imaging with PET. PMID- 18426241 TI - Metals in neurobiology: probing their chemistry and biology with molecular imaging. PMID- 18426243 TI - Strategic biopharmaceutical portfolio development: an analysis of constraint induced implications. AB - Optimizing the structure and development pathway of biopharmaceutical drug portfolios are core concerns to the developer that come with several attached complexities. These include strategic decisions for the choice of drugs, the scheduling of critical activities, and the possible involvement of third parties for development and manufacturing at various stages for each drug. Additional complexities that must be considered include the impact of making such decisions in an uncertain environment. Presented here is the development of a stochastic multi-objective optimization framework designed to address these issues. The framework harnesses the ability of Bayesian networks to characterize the probabilistic structure of superior decisions via machine learning and evolve them to multi-objective optimality. Case studies that entailed three- and five drug portfolios alongside a range of cash flow constraints were constructed to derive insight from the framework where results demonstrate that a variety of options exist for formulating nondominated strategies in the objective space considered, giving the manufacturer a range of pursuable options. In all cases limitations on cash flow reduce the potential for generating profits for a given probability of success. For the sizes of portfolio considered, results suggest that naively applying strategies optimal for a particular size of portfolio to a portfolio of another size is inappropriate. For the five-drug portfolio the most preferred means for development across the set of optimized strategies is to fully integrate development and commercial activities in-house. For the three drug portfolio, the preferred means of development involves a mixture of in house, outsourced, and partnered activities. Also, the size of the portfolio appears to have a larger impact on strategy and the quality of objectives than the magnitude of cash flow constraint. PMID- 18426242 TI - Site-specific linking of biomolecules via glycan residues using glycosyltransferases. AB - The structural information on glycosyltransferases has revealed that the sugar donor specificity of these enzymes can be broadened to include modified sugars with a chemical handle that can be utilized for conjugation chemistry. Substitution of Tyr289 to Leu in the catalytic pocket of bovine beta-1,4 galactosyltransferase generates a novel glycosyltransferase that can transfer not only Gal but also GalNAc or a C2-modified galactose that has a chemical handle, from the corresponding UDP-derivatives, to the non-reducing end GlcNAc residue of a glycoconjugate. Similarly, the wild-type polypeptide-N-acetyl galactosaminyltransferase, which naturally transfers GalNAc from UDP-GalNAc, can also transfer C2-modified galactose with a chemical handle from its UDP derivative to the Ser/Thr residue of a polypeptide acceptor substrate that is tagged as a fusion peptide to a non-glycoprotein. The potential of wild-type and mutant glycosyltransferases to produce glycoconjugates carrying sugar moieties with chemical handle makes it possible to conjugate biomolecules with orthogonal reacting groups at specific sites. This methodology assists in the assembly of bio-nanoparticles that are useful for developing targeted drug-delivery systems and contrast agents for magnetic resonance imaging. PMID- 18426244 TI - Cleaning characterization of protein drug products using UV-vis spectroscopy. AB - This study uses on-line absorbance monitoring to evaluate cleanability of protein drug products. Characterization and validation of equipment cleanliness is a key requirement for a biopharmaceutical facility. A manufacturing-scale cleaning cycle has to be developed and validated for its ability to clean all of the equipment parts for a given soil. Cleaning validation in a multiproduct fill finish facility could benefit from using a worst-case-based approach that involves validating the cleaning process for the most difficult to clean product. Such an approach minimizes the number of required validation runs. Scaled-down cleaning evaluations can provide helpful information for evaluating multiple products and determine the worst case. This study presents a simple and rapid technique for bench-scale characterization of cleanability of protein drug products. On-line A280 (UV absorbance at 280 nm) measurements are performed using a fiber optic probe, and the data are used to establish the dynamics of protein dissolution in cleaning solution. The model not only helps to estimate cleaning time of different formulated proteins (and peptides) but also provides insights into the kinetics of cleaning under different thermal and chemical conditions. Protein product degradation during cleaning is also evaluated through gel electrophoresis. Such information is useful in designing new cleaning cycles. While the study is performed using drug products, the model as well as the findings are also applicable for characterization of final purified bulk soils relevant to bulk drug manufacturing. PMID- 18426246 TI - "I didn't think I was going to like working with him, but now I really do!": examining peer narratives of significant disability. AB - Mainstream research that examines relations between students with significant disabilities and their peers continues to assess such relations on the capacity of students with significant disabilities to evoke and sustain them. This article adopts a disability studies approach to situate peer relations within the larger classroom context. The author draws on the data collected from a qualitative study that investigated the participation of a student with significant disabilities, Harry (a pseudonym), in an inclusive 1st-grade classroom. The author describes peer relations with Harry as embedded within the paradigmatic "family" narrative within this setting. Despite its benefits, the adherence to a normative framework within this family narrative constrained Harry's participation and the kinds of relations that evolved between him and his peers. PMID- 18426245 TI - Advances in cell culture process development: tools and techniques for improving cell line development and process optimization. AB - At the 234th National Meeting of the American Chemical Society, held in Boston, MA, August 19-23, 2007, the ACS BIOT division held two oral sessions on Cell Culture Process Development. In addition, a number of posters were presented in this area. The critical issues facing cell culture process development today are how to effectively respond to the increase in product demands and decreased process timelines while maintaining robust process performance and product quality and responding to the Quality by Design initiative promulgated by the Food and Drug Administration. Two main areas were addressed in the presentations: first, to understand the effects of process conditions on productivity and product quality, and second, to achieve improved production cell lines. A variety of techniques to achieve these goals were presented, including automated flow cytometric analysis, a high-throughput cell analysis and selection method, transcriptional and epigenetic techniques for analysis of cell lines and cell culture systems, and novel techniques for glycoform analysis. PMID- 18426247 TI - The stability of IQ in people with low intellectual ability: an analysis of the literature. AB - A meta-analysis of the stability of low IQ (IQ < 80) was performed on IQ tests that have been commonly used--tests that were derived by D. Wechsler (1949, 1955, 1974, 1981, 1991, 1997) and those based on the Binet scales (L. M. Terman, 1960; L. M. Terman & Merrill, 1972). Weighted-mean stability coefficients of .77 and .78 were found for Verbal IQ (V IQ) and Performance IQ (P IQ) on the Wechsler tests and .82 for Full-Scale IQ (FS IQ) on both Wechsler and Binet tests, for a mean test-retest interval of 2.8 years. Although the majority of FS IQs changed by less than 6 points, 14% changed by 10 points or more. The author suggests that the results of IQ assessment should be treated with more caution than previously thought. PMID- 18426248 TI - Asian American parents' attributions of children with Down syndrome: connections with child characteristics and culture. AB - This study explores cultural differences between European American (n = 26) and Asian American (n = 17) parents' attributional ratings of children with Down syndrome. Links were examined among parents' attributions, reactions, and behaviors regarding their child's jigsaw-puzzle performance. Although the children's puzzle abilities did not differ, compared with European American parents, Asian American parents judged their child as less successful and had lower expectations for future success. Asian American parents also attributed the child's performance to lower ability and lower effort. Affectively, they indicated less sympathy and more anger and blame toward the child. Despite striking ethnic differences, parents in both groups judged their older children as more successful and reported offering them less encouragement and help. Implications of these findings are discussed. PMID- 18426250 TI - Trading one myth for another? With apologies to Dr. Brabner. PMID- 18426251 TI - Misuse of the Street Survival Skills Questionnaire (SSSQ) for evaluating the adult adaptive behavior of criminal defendants with intellectual disability claims. PMID- 18426252 TI - Current practices and future directions of special education in the Republic of Korea. PMID- 18426253 TI - Human services restraint: its past and future. PMID- 18426254 TI - Participation in integrated employment and community-based nonwork services for individuals supported by state disability agencies. PMID- 18426257 TI - Family decision making: benefits to persons with developmental disabilities and their family members. AB - Family involvement in planning and choosing services has become a key intervention concept in developmental disability services. This study (N = 547) modeled patterns of family decision making and assessed benefits to persons with developmental disabilities (DDs) and their family members. A latent profile analysis identified 4 classes that were highly involved in decision making (n = 118), involved only in planning (n = 166), involved only in financial decisions (n = 75), and uninvolved (n = 188). Multiple regression analysis indicated that consumers with DD whose family members were highly involved received more services than consumers in other families. A multivariate analysis of covariance indicated that the family members in the highly involved and planning classes experienced more family member satisfaction than others. Findings have implications for practice. PMID- 18426258 TI - A survey on the impact of being depressed on the professional status and mental health care of physicians. AB - CONTEXT: Recent studies have addressed the need to better understand the nature and risk of depression and suicide in physicians. OBJECTIVE: To assess the prevalence of depressive symptoms in a sample of practicing physicians, their perceptions of the impact of depression on their work lives, and their perceptions of the impact of being a physician on their pursuit of mental health care. DESIGN: An anonymous survey was mailed in April 2005 that included the Patient Health Questionnaire depression module (PHQ-9) and other Likert-style questions. PARTICIPANTS: Five thousand randomly selected practicing physicians in Michigan, from whom 1154 usable responses were received (23% response rate). MAIN OUTCOME MEASURES: The prevalence of depressive symptoms and the perceptions by respondents of the impact of depression on work roles and on their approach to seeking mental health care. RESULTS: Moderate to severe depression scores were reported by 130 physicians (11.3%). Roughly one quarter of respondents reported knowing a physician whose professional standing had been compromised by being depressed. Physicians reporting moderate to severe depression were 2 to 3 times more likely to report substantial impact on their work roles compared to physicians with minimal to mild depression scores, including a decrease in work productivity (57.7% vs. 18.5%; p < .001) and a decrease in work satisfaction (90.8% vs. 36.2%; p < .001). The same physicians were 2 to 3 times more likely to report a wide range of dysfunctional and worrisome approaches to seeking mental health care compared to physicians with minimal to mild depression scores, including a higher likelihood that they would self-prescribe antidepressants (30.0% vs. 9.9%; p < .001) and a higher likelihood that they would avoid seeking treatment due to concerns about confidentiality (50.7% vs. 17.3%; p < .001). CONCLUSIONS: Moderate to severe depression scores are reported by a substantial portion of practicing physicians in Michigan, with important influences on physician work roles and potential negative impact on licensing and medical staff status. The risk of being stigmatized may cause depressed physicians to alter their approach to seeking mental health care, including seeking care outside their medical community and self-prescribing antidepressants. Destigmatization of depression in physicians and interventions to improve the mental health care of physicians in ways that do not compromise their professional standing should receive more attention. PMID- 18426259 TI - Prevalence, correlates, disability, and comorbidity of DSM-IV borderline personality disorder: results from the Wave 2 National Epidemiologic Survey on Alcohol and Related Conditions. AB - OBJECTIVES: To present nationally representative findings on prevalence, sociodemographic correlates, disability, and comorbidity of borderline personality disorder (BPD) among men and women. METHOD: Face-to-face interviews were conducted with 34,653 adults participating in the 2004-2005 Wave 2 National Epidemiologic Survey on Alcohol and Related Conditions. Personality disorder diagnoses were made using the Wave 2 Alcohol Use Disorder and Associated Disabilities Interview Schedule-DSM-IV Version. RESULTS: Prevalence of lifetime BPD was 5.9% (99% CI = 5.4 to 6.4). There were no differences in the rates of BPD among men (5.6%, 99% CI = 5.0 to 6.2) and women (6.2%, 99% CI = 5.6 to 6.9). BPD was more prevalent among Native American men, younger and separated/divorced/widowed adults, and those with lower incomes and education and was less prevalent among Hispanic men and women and Asian women. BPD was associated with substantial mental and physical disability, especially among women. High co-occurrence rates of mood and anxiety disorders with BPD were similar. With additional comorbidity controlled for, associations with bipolar disorder and schizotypal and narcissistic personality disorders remained strong and significant (odds ratios > or = 4.3). Associations of BPD with other specific disorders were no longer significant or were considerably weakened. CONCLUSIONS: BPD is much more prevalent in the general population than previously recognized, is equally prevalent among men and women, and is associated with considerable mental and physical disability, especially among women. Unique and common factors may differentially contribute to disorder-specific comorbidity with BPD, and some of these associations appear to be sex-specific. There is a need for future epidemiologic, clinical, and genetically informed studies to identify unique and common factors that underlie disorder-specific comorbidity with BPD. Important sex differences observed in rates of BPD and associations with BPD can inform more focused, hypothesis-driven investigations of these factors. PMID- 18426261 TI - Orlistat in clozapine- or olanzapine-treated patients with overweight or obesity: a 16-week randomized, double-blind, placebo-controlled trial. AB - OBJECTIVE: Undesirable metabolic effects of modern antipsychotics, especially clozapine and olanzapine, merit development of new weight-control strategies, including pharmacologic ones. We investigated the feasibility of treatment with orlistat, a weight-control drug with no central effects, for overweight/obesity in clozapine- or olanzapine-treated male and female patients. METHOD: Add-on orlistat was prescribed for 16 weeks in a randomized, double-blind, placebo controlled clinical trial to patients who were receiving stable clozapine or olanzapine medication and were aged 18 to 65 years, with no compliance with nonpharmacologic programs or hypocaloric diet required. The primary efficacy variable was body weight change. The study was conducted from 2004 through 2005. RESULTS: Of 71 randomly assigned subjects, 63 were eligible for modified intent to-treat analysis. While no statistically significant effect was observed in the whole population, male (but not female) patients benefited from treatment with orlistat (-2.36 kg vs. 0.62 kg on placebo, p = .011). There were 5 responders (16.1%) (those with >or= 5% weight loss) that received orlistat versus 2 responders (6.3%) that received placebo (number needed to treat = 11), but the difference was not statistically significant. CONCLUSIONS: Without a hypocaloric diet, the effect of orlistat in overweight/obese clozapine-or olanzapine-treated patients is modest and may only be seen in men. More studies should define the optimal length of treatment and feasibility of combination of orlistat with behavioral programs in this population. PMID- 18426260 TI - Changes in antidepressant metabolism and dosing across pregnancy and early postpartum. AB - OBJECTIVE: Little information about the disposition of individual antidepressant drugs during pregnancy has been published. We examined the dose requirements and level-to-dose (L/D) ratios of citalopram, escitalopram, and sertraline during pregnancy and after birth. METHOD: Women aged from 32 to 43 years with major depressive disorder according to the Structured Clinical Interview for DSM-IV Axis I Disorders participated in the study. Doses were charted across each week of gestation and post-partum. Samples were collected at 20, 30, and 36 weeks' gestation; delivery; and at 2 and 12 weeks postpartum. Plasma trough levels were obtained 8 to 15 hours after dose intake. Across pregnancy and postpartum, the mean dose-corrected plasma concentrations (L/D ratios) of S- and R-citalopram and S-sertraline, and the corresponding primary chiral metabolites S- and R desmethylcitalopram and N-desmethylsertra-line were assessed. The samples were analyzed for concentrations of stereospecific parent drug and metabolites. The study was conducted from 2003 to 2006. RESULTS: Three women received citalopram, 2 women were treated with escitalopram, and 6 women received sertraline. In 4 of 5 subjects who received citalopram or escitalopram and 5 of 6 subjects who received sertraline, the L/D ratios for the stereoisomers of the parent compound and primary metabolite decreased between 20 weeks gestation and delivery, which reflects increased drug metabolism. By 12 weeks postpartum the L/D ratios were similar to those detected at 20 weeks gestation. CONCLUSIONS: Our cases illustrate that dose requirements frequently increase during the second half of pregnancy to offset increased drug turnover and maintain optimal pharmacotherapy. These findings replicate and extend earlier published data with other antidepressants. TRIAL REGISTRATION: clinicaltrials.gov Identifier: NCT00279370. PMID- 18426262 TI - Depression comorbidity and antidepressant use in veterans with chronic hepatitis C: results from a retrospective chart review. AB - BACKGROUND: The 2002 National Institutes of Health Consensus Conference Statement recommended that both clinical and research efforts be made to increase the availability of hepatitis C virus (HCV) treatment to patients who were previously ineligible because of comorbid psychiatric illness and substance use disorders. However, little research on patients with HCV and comorbid depression has been conducted that can serve to inform and guide treatment of HCV. In this study we characterize the prevalence and severity of co-morbid depression, as well as antidepressant and other psychotropic prescribing patterns, in a sample of U.S. veterans with HCV. METHOD: Participants were recruited between November 2002 and July 2005 from the liver specialty clinic and from a 1-time HCV patient education class conducted through the Portland Department of Veterans Affairs Northwest Hepatitis C Resource Center. Patients who signed informed consent were asked to complete the Beck Depression Inventory, Second Edition (BDI-II), and their medical records were reviewed for information regarding active prescriptions for psychotropic medications and prior psychiatric diagnoses. RESULTS: Of the 881 veterans enrolled in the study, 783 (89%) completed the BDI-II. Approximately one third (34%, 264/783) of the veterans endorsed moderate to severe symptoms of depression (BDI-II score >or= 20), and 37% (290/783) were prescribed an antidepressant; however, 48% (140/290) of veterans prescribed an antidepressant continued to endorse moderate to severe depressive symptoms. Furthermore, of all veterans endorsing moderate to severe symptoms of depression (N = 264), only about half (56%, 148/264) were prescribed an antidepressant. CONCLUSION: On the basis of BDI-II scores, a significant proportion of veterans with HCV experience moderate to severe depressive symptoms. Although antidepressants were the most commonly prescribed psychotropic medication, many who were prescribed an antidepressant continued to experience high levels of depressive symptoms, an important consideration when deciding whether to initiate antiviral therapy to treat HCV. PMID- 18426263 TI - Preliminary experience with cutting balloon angioplasty for iliac artery in-stent restenosis. AB - PURPOSE: To report our preliminary experience using cutting balloon angioplasty (CBA) in symptomatic iliac artery in-stent restenosis (ISR). METHODS: Fourteen cases of hemodynamically significant iliac artery ISR (4 common and 10 external) were treated in 12 men (mean age 64 years, range 55-75). Of the 14 stents involved, 8 were balloon-expandable models and 6 were self-expanding. All patients had symptomatic deterioration of at least 1 clinical category over an average period of 50.2 months (range 6-120) post stenting. The mean length of ISR was 11.9 mm (range 2-48), and the average stenosis was 75.4% (range 52%-98%). Nine ISR lesions were focal (<10 mm), 4 were diffuse (>10 mm), and 1 extended outside the stent margins. RESULTS: CBA was performed after conventional angioplasty failure in 7 lesions and as a primary treatment method in 7 lesions. Single (9 focal lesions) or multiple overlapping (5 diffuse or proliferative lesions) inflations were performed using 6-x10-mm (1 lesion), 7-x10-mm (3 lesions), and 8-x10-mm (10 lesions) devices. There was 1 contained rupture treated with a covered stent. In the remainder of the cases, the cutting balloons allowed successful treatment without further stent implantation. During a mean follow-up of 23.6 months (range 12-60), no patient showed clinical deterioration, and no recurrent ISR was detected with color duplex. CONCLUSION: CBA shows high immediate technical and midterm clinical success in symptomatic iliac artery ISR. PMID- 18426264 TI - Risk factors for in-stent restenosis after vertebral ostium stenting. AB - PURPOSE: To determine whether vascular risk factors, underlying vessel diameter, and/or the type of stent affect restenosis rates for vertebral ostium stents. METHODS: A single-center retrospective analysis was conducted of 44 patients (31 men; mean age 61 years, range 32-81) who underwent stenting of 48 ostial lesions in the vertebral arteries between 1999 and 2005. Only patients who underwent angiographic follow-up were included in the analysis. Cox regression analysis was utilized for risk factor association with binary restenosis (> or =50% versus <50%). Stent types and stent categories were compared for differences in binary restenosis rates and lumen gain at follow-up angiography. RESULTS: Twenty-three (48%) of 48 lesions had > or =50% stenosis at a mean follow-up of 7.7 months. Cigarette smoking was associated with higher binary restenosis rates (p=0.025), while hypertension, diabetes, hyperlipidemia, history of neck radiation, and known coronary artery and/or peripheral vascular disease were not. Reduced binary restenosis rates and improved lumen gain were seen in cobalt chromium balloon expandable stents compared to non-cobalt chromium stents (p=0.002 and p=0.002, respectively), stainless steel balloon-expandable stents (p=0.005 and p=0.005), and the S670 stent (p=0.069 and p=0.069). The size of stent used was not associated with risk of restenosis (p=0.756). CONCLUSIONS: Cobalt chromium stents were associated with reduced restenosis, while smoking was associated with increased restenosis risk. PMID- 18426265 TI - Treatment of renal artery aneurysm with the multilayer stent. AB - PURPOSE: To describe a new type of stent consisting of a 3-dimensional (3D) braided tube made of 2 interconnected layers without any covering to treat a renal artery aneurysm. CASE REPORT: A 78-year-old hypertensive man with multiple comorbidities was incidentally found to have a large (28- x 30 mm) saccular aneurysm in the main right renal artery involving the inferior renal artery. Via a percutaneous femoral approach, a 6- x 30-mm Multilayer stent was deployed easily in front of the aneurysm neck covering the inferior renal artery. Blood flow inside the sac was immediately and significantly reduced. All the renal artery branches remained patent. Blood pressure returned to normal after the procedure. At 6 months, angiography showed complete shrinkage of the aneurysm wall; all the inferior renal artery branches remained patent. CONCLUSION: The 3D multilayer fluid modulating stent concept appears to be a viable alternative for renal aneurysm exclusion. A larger study is underway to evaluate this new stent in other peripheral aneurysms. PMID- 18426266 TI - Late complication of stent fragmentation related to the "lever-arm effect". AB - PURPOSE: To report an unusual case of dramatic nitinol stent strut fractures with resultant complete loss of stent integrity in a cadaveric lower limb graft. CASE REPORT: A 73-year-old man had the clinical stigmata of graft versus host response to a cadaveric femoral-tibioperoneal graft and suspected inflammation-induced stent corrosion culminating in fracture. The long continuous stented segment and station of the distal anastomosis below the knee resulted in preferential displacement of force to the proximal stent edge as the knee bends, making mechanical stress the most likely primary cause for these findings. CONCLUSION: This case provides insight into some of the unique mechanical issues in vessels traversing the knee joint and how the problem is amplified by increasing the distance between the knee (fulcrum) and the stented vessel (lever arm). This lever-arm effect would also apply to the native superficial femoral artery when vessel compliance is changed by multiple stents. PMID- 18426267 TI - Multicenter experience with a new "hybrid" carotid stent. AB - PURPOSE: To report the results from a prospective multicenter registry evaluating the safety and performance of a new hybrid stent specifically engineered for carotid indications. METHODS: Between October 18, 2006, and March 28, 2007, 124 patients (88 men; mean age 71.8+/-7.3 years, range 52-87) underwent Cristallo Ideale stent implantation at 4 expert sites in Italy and Germany. Eligibility criteria included >60% symptomatic and >80% asymptomatic carotid stenosis. Different cerebral protection devices were utilized (proximal protection and distal filters). The primary endpoint was the rate of 30-day major adverse neurological events, defined as major and minor stroke and neurological death by independent neurological assessment. All adverse events were adjudicated by an independent clinical event committee. RESULTS: Stent deployment was completed successfully in all the cases without documented technical failure. There were no neurological events during the procedures. Thirty-day follow-up was available for 119 (96.0%) patients [2 (1.6%) were lost to follow-up and 3 (2.4%) died from non neurological causes unrelated to the device or procedure]. There were no major adverse neurological events within 30 days, but 2 (1.6%) device-/procedure related transient ischemic attacks were reported (both resolved completely in <24 hours). Four (3.2%) other non-neurological events (anemia requiring transfusion, worsening of pre-existing chronic renal failure, acute access site thrombosis, and monocular vision disturbance) were reported in the study period. CONCLUSION: The Cristallo Ideale carotid stent is safe and effective in the periprocedural period. Its newly designed hybrid structure seems to support the rationale of combining adequate plaque scaffolding with high vessel adaptability. PMID- 18426268 TI - Wall apposition assessment and performance comparison of distal protection filters. AB - PURPOSE: To assess the wall apposition of 3 distal protection filters used in carotid artery stenting (CAS) for cerebral protection and quantify the effect on the in vitro capture efficiency of the filters under simulated physiological flow conditions. METHODS: The 3 distal protection filters (Angioguard XP, FilterWire EZ, and RX Accunet) were deployed in silicone flow models of 5.0-, 5.5-, and 6.0 mm inner diameter and were tested at a mean flow rate characteristic of the human internal carotid artery while injecting polydispersed microspheres simulating plaque emboli. The injected microspheres had a diameter larger than the pore size of the devices tested, so it was conjectured that any microspheres missed by the device traveled between the device basket and the vessel wall. Varying the diameter of the vessel phantom within the recommended vessel diameter treatment range for each device simulated the variability of vessel diameter in vivo, allowing the quantification of device wall apposition. RESULTS: None of the devices tested completely prevented distal embolization in the flow model. The RX Accunet device has the best overall wall apposition, yielding gaps of 0.075% of the vessel cross-sectional area. The FilterWire EZ device had the best overall average filtration rate, failing to capture only 0.8% of plaque particles. There were no statistically significant differences in the wall apposition assessment or the capture efficiency of the RX Accunet and FilterWire EZ devices. CONCLUSION: Several complications related to apposition of the filter basket on the vessel wall and device retrieval were detected in all the devices. It is inferred that the adaptability of the filter basket to conform to the vessel cross section at the site of deployment is the primary design variable responsible for distal embolization during CAS with cerebral protection. PMID- 18426269 TI - Clinical implications of systemic inflammatory response syndrome following thoracic aortic stent-graft placement. AB - PURPOSE: To quantify inflammatory markers in a large cohort of patients undergoing thoracic endovascular aortic repair (TEVAR) and investigate if profound biomarker elevations may be predictors of postprocedural death. METHODS: We analyzed data from 103 patients (70 men; mean age 64.5+/-11.2 years, range 22 83) undergoing TEVAR between July 1999 and December 2006. Baseline as well as at least 3 serial measurements of C-reactive protein (CRP), fibrinogen, white blood cell (WBC) count, and D-dimers were performed within the first 20 days after TEVAR. RESULTS: Compared with baseline, all inflammatory biomarker levels rose significantly. WBC peaked 2 days after the procedure, whereas CRP, fibrinogen, and D-dimers showed a sustained elevation up to 20 days after TEVAR. Inflammatory responses were more pronounced in patients with acute aortic pathology compared with chronic aortic diseases. There was evidence of greater increase in biomarkers with an increasing number of stent-grafts implanted. Kaplan-Meier analysis suggested that increasing maximum D-dimer values postoperatively were associated with decreased survival after TEVAR (p=0.036) in a subset of patients; however, multivariate analysis failed to identify postinterventional biomarker elevation as independent predictor of in-hospital death. CONCLUSION: Postprocedural inflammatory responses characterized by elevations of CRP, fibrinogen, D-dimers, and WBC are observed in all patients undergoing TEVAR. Our data indicate that this response is more pronounced in patients with acute aortic pathology and those receiving >1 stent-graft. PMID- 18426270 TI - Thoracic aortic endografting in patients with connective tissue diseases. AB - PURPOSE: To present midterm results after thoracic endovascular aortic repair (TEVAR) in patients with connective tissue diseases focusing on secondary endoleak and reintervention due to disease progression. METHODS: Between January 1997 and January 2007, 167 patients received 241 thoracic aortic stent-grafts. Eight patients (6 men; median age 48 years, range 32-67) with connective tissue diseases (6 Marfan and 2 Ehlers-Danlos syndrome) treated with stent-graft repair were retrospectively analyzed at a median follow-up of 31 months (range 3-79). Surveillance included postoperative computed tomographic angiography and/or magnetic resonance imaging exams prior to discharge, at 3, 6, and 12 months, and yearly thereafter. RESULTS: Technical success of endovascular placement was 88% due to 1 primary type I endoleak. There were no perioperative deaths, and there have been no conversions to open surgery so far. Perioperative complications occurred in 2 (25%) of the 8 patients. Endoleaks were observed in 3 patients (primary type I, secondary type I, and type II). The reintervention rate was 38%. Progression of disease resulting in de novo aneurysms or aortic expansion occurred in 4 (50%) patients. Seven (88%) patients are alive. There was no disease- or procedure-related death. CONCLUSION: TEVAR in patients with connective tissue diseases is feasible but still questionable regarding their young age and the rates of endoleaks and reintervention due to disease progression. Close surveillance is mandatory. Low morbidity and mortality rates may justify TEVAR in emergencies as a "bridging" method. PMID- 18426271 TI - Meta-analysis of endovenous radiofrequency obliteration of the great saphenous vein in primary varicosis. AB - PURPOSE: To compare radiofrequency obliteration (RFO) and conventional surgery with respect to postoperative complications, effectiveness of treatment, and quality of life (QoL). METHODS: Several healthcare databases were interrogated to identify all studies published between 1994 and 2007 comparing RFO in primary varicosis to conventional therapy with vein ligation and stripping. Of 65 articles identified, 8 studies representing 428 patients [224 (52%) endovenous RFO and 204 (48%) stripping] were eligible for the meta-analysis. Adverse events, effectiveness, and QoL outcomes were assessed at several time points up to 2 years. RESULTS: There were significant reductions in tenderness and ecchymosis at 1 week and significantly fewer hematomas at 72 hours, 1 week, and 3 weeks associated with RFO. There was no significant difference between the RFO and surgery in immediate or complete great saphenous vein (GSV) occlusion, incomplete GSV closure, freedom from reflux, recurrent varicose veins, recanalization, or neovascularization. QoL results significantly favoring RFO over surgery included return to normal activity and return to work. CONCLUSION: It seems that RFO benefits most patients in the short term, but rates of recanalization, re treatment, occlusion, and reflux may alter with longer follow-up. The lack of such data demonstrates the need for further randomized clinical trials of RFO versus conventional surgery. PMID- 18426272 TI - Endovascular repair of acute type B aortic dissection: midterm results. AB - PURPOSE: To evaluate midterm results of endovascular stent-graft placement for acute Stanford type B dissection (TBD). METHODS: A retrospective review was conducted of 35 consecutive patients who were treated with stent-graft implantation for acute TBD between July 1996 and July 2007. Computed tomographic (CT) volumetric analysis of the true lumen (TL) and false lumen (FL) changes in 23 patients was performed, as well as evaluation of the influence of re-entry points and length of stent-graft coverage on volume changes. In addition, complications were evaluated. Follow-up was performed at 6 and 12 months and yearly thereafter. RESULTS: The technical success rate was 82.7%, and the 30-day mortality rate was 8.5%. Mean follow-up was 34 months. The overall survival rate at 5 years was 78.4%. Complications included retrograde type A dissections in 3 patients during the perioperative period and in 1 patient during midterm follow up. In addition, 5 early and 3 late endoleaks were observed. Three patients were converted to open surgery and 2 needed secondary interventions. In the stented segment, stabilization of the aorta was achieved even during midterm follow-up, with a TL volume increase of 59% at 5 years and nearly stable FL volume. The segment from the distal end of the stent-graft to the celiac artery, however, showed unstable TL and FL volumes, with high standard deviations after the first postinterventional year and circumferential aneurysmal dilatation of the aorta immediately adjacent to the stent-graft in 5 patients. The abdominal aorta showed no substantial volume changes over time. The length of stent-graft coverage and the occurrence of re-entries greatly influenced FL volume changes distal to the stent-graft. CONCLUSION: Serious complications can occur during and after endovascular repair of TBD. Therefore, it should be reserved for high-risk patients. PMID- 18426273 TI - Retrograde stenting of a recurrent degenerative aneurysm of the superior mesenteric artery. PMID- 18426274 TI - Double-barrel technique for endovascular exclusion of an aortic arch aneurysm without sternotomy. AB - PURPOSE: To report the use of commercially available stents and a stent-graft in coaxial orientation to extend the proximal limits of endografting within the aortic arch. CASE REPORT: A 70-year-old man was found to have an asymptomatic 7 cm saccular aortic arch aneurysm, extending distally from the origin of the left carotid artery and involving the left subclavian artery; there was only 11 mm between the innominate artery orifice and the aneurysm. The patient was deemed to be high risk for open surgical repair due to a history of 2 prior sternotomies and the requirement for hypothermic circulatory arrest. A "double-barrel" stent graft strategy combining retrograde placement of an innominate stent with thoracic stent-graft implantation into zone 0 was successfully executed. The patient has continued to fare well after 10 months on close follow-up. CONCLUSION: The "double-barrel" stent technique may extend the limits of thoracic endografting by preserving the aortic arch branches while avoiding the need for sternotomy. Using this technique, proximal fixation can be obtained well into the ascending aorta using commercially available devices. PMID- 18426275 TI - Commentary: conquering zone zero: expanding endograft repair in the aortic arch. PMID- 18426276 TI - Endovascular aortic occlusion for a secondary aorto-enterocutaneous fistula: 44 month follow-up. AB - PURPOSE: To describe the durability of endovascular aortic occlusion as an alternative treatment of an aortoenteric fistula in a severely unstable patient. CASE REPORT: A 47-year-old patient with a history of numerous previous vascular reconstructions underwent surgical exclusion of a persistent aorto enterocutaneous fistula originating from a former colostomy site in proximity with the occluded left leg of an aortobifemoral bypass. Nine weeks later, he presented in profound hypovolemic shock from bleeding via the cutaneous remnant of the fistula and gastrointestinal tract. Because of the severe shock, recent surgery, and lack of a suitable endograft for the small-caliber aorta (19 mm), endovascular exclusion of the fistula was undertaken by occluding the infrarenal aorta. Talent occluders were also placed in the right femoral bypass limb and the right common iliac artery. An extra-anatomical bypass was created to revascularize the lower extremities. The patient was discharged in good condition on day 20. The fistula spontaneously regressed after 12 months. Since the patient refused further surgery, 44-month follow-up is now available. There are no signs of reinfection or recurrent aortoenteric fistula. CONCLUSION: Endovascular occlusion of a small caliber aorta with extra-anatomical revascularization of the lower extremities may be considered a useful treatment alternative for an aortoenteric fistula in severely unstable or compromised patients, either as a bridge to open surgery or as a definitive treatment if signs of reinfection do not occur. PMID- 18426277 TI - Histological diagnosis of atypical Takayasu arteritis using percutaneous transluminal atherectomy. AB - PURPOSE: To describe the use of percutaneous directional atherectomy to aid in the diagnosis of atypical Takayasu arteritis in the lower limb vessels. CASE REPORT: In a 24-year-old woman with severe peripheral artery disease (Rutherford Becker category 5), imaging showed bilateral occlusion of the superficial femoral arteries (SFA), but no lesions of the proximal or supra-aortic vessels. The right SFA was treated first; after guidewire passage through the occlusion, 7 passes of the SilverHawk Peripheral Atherectomy Catheter debulked the lesion, followed by balloon angioplasty. Catheter biopsy showed inflammatory infiltrates consistent with Takayasu arteritis. CONCLUSION: Percutaneous transluminal atherectomy is not only an interventional device but also a diagnostic tool and can be used to distinguish between inflammatory and atherosclerotic lesions. PMID- 18426278 TI - Late iliac artery aneurysm formation: implications for the lower landing site after EVAR. PMID- 18426279 TI - Management of the left subclavian artery during endovascular repair of the thoracic aorta. AB - Endovascular repair is rapidly becoming the treatment of choice for thoracic aortic disease, which oftentimes involves or lies in close proximity to the left subclavian artery (LSA). In order to extend the proximal landing zone for the stent-graft and obtain an adequate seal, the LSA ostium is often covered, with or without concomitant subclavian artery revascularization. In this article, we review the LSA anatomy and consequences of LSA coverage as a backdrop for a discussion of the ramifications of LSA coverage during endovascular thoracic aortic repair (TEVAR). Early series reported high rates of LSA revascularization as an adjunct to endovascular repair for aortic pathology adjacent to the LSA ostium. Initial reports of low morbidity associated with simple LSA ostium coverage are not supported by contemporary literature, which suggests revascularization reduces the risks of cerebrovascular accident and spinal cord ischemia. Coverage of the LSA without revascularization may be justified only in emergency situations or when thorough investigations of cerebral and vertebrobasilar circulation have concluded that the risk to brain and spinal cord is low. Subclavian revascularization should be considered in the presence of a dominant left vertebral artery, bilateral carotid artery disease, an occluded/stenosed right vertebral artery, presence of a left internal mammary artery graft, or when a long length of thoracic aorta is covered. PMID- 18426280 TI - Expressed emotion, communication deviance, and culture in families of patients with schizophrenia: a review of the literature. AB - The aim of this article is to critically review the literature on expressed emotion (EE), communication deviance (CD), and culture in families of patients with schizophrenia. There is growing evidence that EE and CD are highly linked. Yet the two constructs together predict the development of schizophrenia and the associated symptoms better than either construct alone. In this article, the authors review data indicating that both the expression and the levels of high EE and CD vary by ethnicity. It may be especially difficult for family members to communicate coherently and in a less critical manner when focusing on patients' inability to sustain particular cultural norms and values that are endorsed by their family and ethnic background. The authors propose that more attention to the role of culture in EE and CD and greater focus on the proper assessment of these variables would further enhance our understanding of these constructs. PMID- 18426281 TI - Maladaptive perfectionism and depressive symptoms among Asian American college students: contributions of interdependence and parental relations. AB - Maladaptive perfectionism among Asian Americans may represent a shared social construction socialized within an interdependent cultural script. The authors hypothesized that interdependence concerns and parental relations may explain elevated maladaptive perfectionism and associated depressive symptoms in a sample of 140 Asian American college students. Survey findings revealed that interdependence, maladaptive perfectionism, and parent-driven perfectionism were associated with depressive symptoms. Regression analyses revealed that interdependence moderated the relationship between maladaptive perfectionism and depressive symptoms such that highly interdependent Asian American students appeared more vulnerable to depression when demonstrating perfectionistic tendencies. Parental support buffered subjects from distress associated with parent-driven perfectionism. Processes of heightened cultural vulnerability and sensitization to maladaptive perfectionism are discussed. PMID- 18426282 TI - "But you don't look Puerto Rican": the moderating effect of ethnic identity on the relation between skin color and self-esteem among Puerto Rican women. AB - This exploratory study investigated whether ethnic identity, as assessed by Phinney's (1992) Multigroup Ethnic Identity Measure, functioned as a moderator in the relation between skin color (as measured by masked interviewer evaluation, participant self-report, and skin reflectance data) and self-esteem (as measured by Rosenberg's 1989 Self-Esteem Scale). In a sample of 53 English-speaking Puerto Rican women, a hierarchical multiple regression indicated that among lighter skinned women, those who felt less attached to their culture had less self-esteem than those who were more culturally embedded. Similarly, among darker skinned women, greater attachment to Puerto Rican culture was associated with greater self-esteem than a less defined ethnic identity. Findings are discussed in light of the beneficial effects of ethnic identity. PMID- 18426283 TI - Mothers' and fathers' involvement with school-age children's care and academic activities in Navajo Indian families. AB - This exploratory study examined mothers' and fathers' reports of time involvement in their school-age children's care and academic activities. The study also explored the relationship between parents' socioeconomic status (SES) variables (age, education, income, work hours, and length of marriage) and their relative involvement with children. Mother and father dyads from 34 two-parent Navajo (Dine) Indian families with a second- or third-grade child participated in the study. Repeated measures analysis of variance showed that mothers invested significantly more time in children's care on demand and academic activities than fathers, but the differences in maternal and paternal perceptions of time involvement in routine care were not significant. The gender of the child did not influence the amount of time parents invested in children's care and academic activities. Mothers' involvement with children was not related to any of the SES variables. Fathers' involvement was significantly associated with work hours and length of marriage, and work hours produced significant interaction with fathers' involvement with children. Findings are discussed in light of gender role differences in parental involvement with children within Navajo families. PMID- 18426284 TI - A colonial mentality model of depression for Filipino Americans. AB - Many cultural and ethnic minorities have extensive experiences of being oppressed, which they may eventually internalize. However, psychology has yet to actively incorporate various forms of internalized oppression (e.g., colonial mentality [CM]) into the etiological conceptualizations of psychopathology. Using a sample of 248 Filipino Americans, the author tested a more complete and sociopolitically informed cultural model of depression symptoms. Results with structural equation modeling showed that a conceptual model that includes CM better explained depression symptoms among Filipino Americans than the model without CM and revealed that CM had a significant direct effect on Filipino Americans' experiences of depression symptoms. It is argued, through this illustrative case of depression symptoms among Filipino Americans, that incorporating the psychological effects of oppressive historical and contemporary conditions into our conceptualizations of ethnic minority mental health may lead to a more culturally accurate etiological understanding of psychopathology among historically oppressed groups. PMID- 18426285 TI - What it means to be and feel like a "true" American: perceptions and experiences of second-generation Americans. AB - Through semistructured interviews, the authors investigated the relationships between the definition of a "true" American, the impact of 9/11, and the war in Iraq and their impact on experiences of fitting into and being excluded from the American identity with a sample of 10 2nd-generation young adults. Using consensual qualitative research methods (C. E. Hill et al., 2005; C. E. Hill, B. Thompson, & E. N. Williams, 1997) the authors identified 6 major domains that described participants' experiences, including physical characteristics of true Americans, behavioral characteristics of true Americans, beliefs and values of true Americans, the impact of 9/11 on definitions of true Americans, participants' American identity, and experiences as 2nd-generation Americans. PMID- 18426286 TI - Sexual values among Latino youth: measurement development using a culturally based approach. AB - Latino youth in the United States are at higher risk for negative sexual outcomes compared to their European American counterparts. Adherence to traditional sexual values may protect against or increase their risk. Past studies have generally utilized proxy measures, such as acculturation, to assess sexual values. The objective of the current study was to develop and test culturally based sexual values measures among Latino youth. Focus groups and qualitative interviews were conducted to generate themes related to sexual values. Six measures were developed: three related to gender role norms (Sexual Talk as Disrespectful, Satisfaction of Sexual Needs as Important, Female Virginity as Important) and three related to comfort regarding sexuality (Comfort with Sexual Communication, Sexual Comfort, and Sexual Self-Acceptance). The scales' psychometric properties were assessed in a sample of 694 sexually active young Latinos. Results indicate that these measures conformed to single-factor scales and displayed acceptable reliabilities. Correlations with conceptually related measures were in hypothesized directions. Findings suggest it is feasible to directly assess sexual values in a valid and reliable manner. The measures presented in the current article represent tools for such assessment. PMID- 18426287 TI - Disaggregating the effects of acculturation and acculturative stress on the mental health of Asian Americans. AB - This study examines the impact of level of acculturation and acculturative stress on the mental health of Asian American college students. Hierarchical regression analyses were used to clarify the relation between level of acculturation, acculturative stress, and mental health outcomes (psychological distress and clinical depression). Being less identified with mainstream United States culture was associated with higher psychological distress and clinical depression, but lost significance when acculturative stress was introduced into the model. Retention or relinquishing of identification with one's heritage culture was not associated with mental health outcomes. Although understanding level of acculturation can help us identify those at risk, findings suggest that acculturative stress is a more proximal risk factor and increases risk for mental health problems independently of global perceptions of stress. PMID- 18426288 TI - Familialism, social support, and stress: positive implications for pregnant Latinas. AB - This study examined the association of familialism, a cultural value that emphasizes close family relationships, with social support, stress, pregnancy anxiety, and infant birth weight. Foreign-born Latina (n = 31), U.S.-born Latina (n = 68), and European American (n = 166) women living in the United States participated in a prospective study of pregnancy in which they completed measures of familialism, social support, stress, and pregnancy anxiety during their second trimester. As expected, Latinas scored higher on familialism than European Americans. Familialism was positively correlated with social support and negatively correlated with stress and pregnancy anxiety in the overall sample. As predicted, however, the associations of familialism with social support and stress were significantly stronger among Latinas than European Americans. Moreover, higher social support was associated with higher infant birth weight among foreign-born Latinas only. Implications of cultural values for relationships and health are discussed. PMID- 18426289 TI - Latina breast cancer survivors' lived experiences: diagnosis, treatment, and beyond. AB - Latina breast cancer survivors are not benefiting from advances in psychosocial interventions. Despite their greater breast cancer burden, there is a dearth of information about this population. This qualitative study examined the experiences of 18 immigrant Latina breast cancer survivors along the survivorship continuum, from diagnosis to long-term survivorship. The authors conducted separate focus groups with women in the acute, reentry, and long-term survivorship stages. Through grounded theory analysis, the authors uncovered 5 themes of experience: perceptions of psychological well-being, impact of diagnosis, impact of treatment, need for social support, and development of new attitudes. Some themes were applicable across all survivorship stages, whereas others were more salient at a particular stage. Within themes, the authors identified experiences reported in the general literature (i.e., the debilitating effect of chemotherapy) and distinct cultural experiences (i.e., fear that a cancer diagnosis leads to certain death). Results suggest that treatments tailored for each survivorship stage are more likely to be effective than generic interventions. These findings have the potential to lessen health disparities in utilization of psychosocial treatments by Latina breast cancer survivors. PMID- 18426290 TI - Depression in Latino adolescents: a cultural discrepancy perspective. AB - Latino adolescents report high levels of depression compared to other youth, yet little is known about how culture-specific factors contribute to risk (Blazer, Kessler, McGonagle, & Swartz, 1994; Roberts, Roberts, & Chen, 1997; Roberts & Sobhan, 1992; Twenge & Nolen-Hoeksema, 2002). In this study we evaluated the link between cultural discrepancy (i.e., perceived acculturation and gender role disparity between children and their parents) and depression among children of Latino immigrants. Compared to boys, Latina adolescents reported greater differences in traditional gender role beliefs between themselves and their parents and higher levels of depression. Gender role discrepancy was associated with higher youth depression, with this relationship mediated by increases in family dysfunction. Moreover, a moderator analysis suggested that gender role discrepancy effects may be most pronounced for Latina adolescents. Gender role discrepancy was associated with poorer family functioning for girls but not for boys, although the interaction effect was only marginally significant. These preliminary results point to the importance of considering cultural discrepancy as a contributing factor to youth depression. PMID- 18426291 TI - The ABCs of depression: integrating affective, biological, and cognitive models to explain the emergence of the gender difference in depression. AB - In adulthood, twice as many women as men are depressed, a pattern that holds in most nations. In childhood, girls are no more depressed than boys, but more girls than boys are depressed by ages 13 to 15. Although many influences on this emergent gender difference in depression have been proposed, a truly integrated, developmental model is lacking. The authors propose a model that integrates affective (emotional reactivity), biological (genetic vulnerability, pubertal hormones, pubertal timing and development) and cognitive (cognitive style, objectified body consciousness, rumination) factors as vulnerabilities to depression that, in interaction with negative life events, heighten girls' rates of depression beginning in adolescence and account for the gender difference in depression. PMID- 18426292 TI - The self-regulation of automatic associations and behavioral impulses. AB - The distinction between automatic processes and controlled processes is a central organizational theme across areas of psychology. However, this dichotomy conceals important differences among qualitatively different processes that independently contribute to ongoing behavior. The Quadruple process model is a multinomial model that provides quantitative estimates of 4 distinct processes in a single task: the likelihood that an automatic response tendency is activated; the likelihood that a contextually appropriate response can be determined; the likelihood that automatic response tendencies are overcome when necessary; and the likelihood that in the absence of other information, behavior is driven by a general response bias. The model integrates dual-process models from many domains of inquiry and offers a generalized, more nuanced framework of impulse regulation across these domains. The model offers insights into many central questions surrounding the operation and the interaction of automatic and controlled processes. Applications of the model to empirical and theoretical concerns in a variety of areas of psychology are discussed. PMID- 18426294 TI - Shortlist B: a Bayesian model of continuous speech recognition. AB - A Bayesian model of continuous speech recognition is presented. It is based on Shortlist (D. Norris, 1994; D. Norris, J. M. McQueen, A. Cutler, & S. Butterfield, 1997) and shares many of its key assumptions: parallel competitive evaluation of multiple lexical hypotheses, phonologically abstract prelexical and lexical representations, a feedforward architecture with no online feedback, and a lexical segmentation algorithm based on the viability of chunks of the input as possible words. Shortlist B is radically different from its predecessor in two respects. First, whereas Shortlist was a connectionist model based on interactive activation principles, Shortlist B is based on Bayesian principles. Second, the input to Shortlist B is no longer a sequence of discrete phonemes; it is a sequence of multiple phoneme probabilities over 3 time slices per segment, derived from the performance of listeners in a large-scale gating study. Simulations are presented showing that the model can account for key findings: data on the segmentation of continuous speech, word frequency effects, the effects of mispronunciations on word recognition, and evidence on lexical involvement in phonemic decision making. The success of Shortlist B suggests that listeners make optimal Bayesian decisions during spoken-word recognition. PMID- 18426295 TI - An integrated model of choices and response times in absolute identification. AB - Recent theoretical developments in the field of absolute identification have stressed differences between relative and absolute processes, that is, whether stimulus magnitudes are judged relative to a shorter term context provided by recently presented stimuli or a longer term context provided by the entire set of stimuli. The authors developed a model (SAMBA: selective attention, mapping, and ballistic accumulation) that integrates shorter and longer term memory processes and accounts for both the choices made and the associated response time distributions, including sequential effects in each. The model's predictions arise as a consequence of its architecture and require estimation of only a few parameters with values that are consistent across numerous data sets. The authors show that SAMBA provides a quantitative account of benchmark choice phenomena in classical absolute identification experiments and in contemporary data involving both choice and response time. PMID- 18426296 TI - The dimensionality of the remember-know task: a state-trace analysis. AB - This article addresses the issue of whether the remember-know (RK) task is best explained by a single-process or a dual-process model. All single-process models propose that remember and know responses reflect different levels of a single strength-of-evidence dimension. Thus, across conditions in which response criteria are held constant, these models predict that the RK task is unidimensional. Many dual-process models propose that remember and know responses reflect two qualitatively distinct processes underlying recognition memory, often characterized as recollection and familiarity. These models predict that the RK task is bidimensional. Using data from 37 studies, the author conducted a state trace analysis to determine the dimensionality of the RK task. In those studies, non-memory-related differences between conditions were eliminated via decision criteria constrained to be constant across all levels of the independent variables. The results reveal little or no evidence of bidimensionality and lend additional support to the unequal-variance signal detection model. Other arguments supporting a bidimensional interpretation are examined, and the author concludes there is insufficient evidence for the RK task to be used to identify qualitatively different memory components. PMID- 18426293 TI - An integrated process model of stereotype threat effects on performance. AB - Research showing that activation of negative stereotypes can impair the performance of stigmatized individuals on a wide variety of tasks has proliferated. However, a complete understanding of the processes underlying these stereotype threat effects on behavior is still lacking. The authors examine stereotype threat in the context of research on stress arousal, vigilance, working memory, and self-regulation to develop a process model of how negative stereotypes impair performance on cognitive and social tasks that require controlled processing, as well as sensorimotor tasks that require automatic processing. The authors argue that stereotype threat disrupts performance via 3 distinct, yet interrelated, mechanisms: (a) a physiological stress response that directly impairs prefrontal processing, (b) a tendency to actively monitor performance, and (c) efforts to suppress negative thoughts and emotions in the service of self-regulation. These mechanisms combine to consume executive resources needed to perform well on cognitive and social tasks. The active monitoring mechanism disrupts performance on sensorimotor tasks directly. Empirical evidence for these assertions is reviewed, and implications for interventions designed to alleviate stereotype threat are discussed. PMID- 18426298 TI - Potential performance theory (PPT): a general theory of task performance applied to morality. AB - People can use a variety of different strategies to perform tasks and these strategies all have two characteristics in common. First, they can be evaluated in comparison with either an absolute or a relative standard. Second, they can be used at varying levels of consistency. In the present article, the authors develop a general theory of task performance called potential performance theory (PPT) that distinguishes between observed scores and true scores that are corrected for inconsistency (i.e., potential scores). In addition, they argue that any kind of improvement to task performance, whatever it may be, works by influencing either task strategies, which comprise all nonrandom components that are relevant to the task, or the consistency with which strategies are used. In the current study, PPT is used to demonstrate how task strategies and the consistencies with which they are used impact actual performance in the domain of morality. These conclusions are extended to other domains of task performance. PMID- 18426300 TI - New paradoxes of risky decision making. AB - During the last 25 years, prospect theory and its successor, cumulative prospect theory, replaced expected utility as the dominant descriptive theories of risky decision making. Although these models account for the original Allais paradoxes, 11 new paradoxes show where prospect theories lead to self-contradiction or systematic false predictions. The new findings are consistent with and, in several cases, were predicted in advance by simple "configural weight" models in which probability-consequence branches are weighted by a function that depends on branch probability and ranks of consequences on discrete branches. Although they have some similarities to later models called "rank-dependent utility," configural weight models do not satisfy coalescing, the assumption that branches leading to the same consequence can be combined by adding their probabilities. Nor do they satisfy cancellation, the "independence" assumption that branches common to both alternatives can be removed. The transfer of attention exchange model, with parameters estimated from previous data, correctly predicts results with all 11 new paradoxes. Apparently, people do not frame choices as prospects but, instead, as trees with branches. PMID- 18426301 TI - The trouble with overconfidence. AB - The authors present a reconciliation of 3 distinct ways in which the research literature has defined overconfidence: (a) overestimation of one's actual performance, (b) overplacement of one's performance relative to others, and (c) excessive precision in one's beliefs. Experimental evidence shows that reversals of the first 2 (apparent underconfidence), when they occur, tend to be on different types of tasks. On difficult tasks, people overestimate their actual performances but also mistakenly believe that they are worse than others; on easy tasks, people underestimate their actual performances but mistakenly believe they are better than others. The authors offer a straightforward theory that can explain these inconsistencies. Overprecision appears to be more persistent than either of the other 2 types of overconfidence, but its presence reduces the magnitude of both overestimation and overplacement. PMID- 18426302 TI - Hebbian learning of cognitive control: dealing with specific and nonspecific adaptation. AB - The conflict monitoring model of M. M. Botvinick, T. S. Braver, D. M. Barch, C. S. Carter, and J. D. Cohen (2001) triggered several research programs investigating various aspects of cognitive control. One problematic aspect of the Botvinick et al. model is that there is no clear account of how the cognitive system knows where to intervene when conflict is detected. As a result, recent findings of task-specific and context-specific (e.g., item-specific) adaptation are difficult to interpret. The difficulty with item-specific adaptation was recently pointed out by C. Blais, S. Robidoux, E. F. Risko, and D. Besner (2007), who proposed an alternative model that could account for this. However, the same problem of where the cognitive system should intervene resurfaces in a different shape in this model, and it has difficulty in accounting for the Gratton effect, a hallmark item-nonspecific effect. The authors of the current article show how these problems can be solved when cognitive control is implemented as a conflict modulated Hebbian learning rule. PMID- 18426303 TI - Is the number of trials a primary determinant of conditioned responding? AB - Acquisition of conditioned responding is thought to be determined by the number of pairings of a conditioned stimulus (CS) and an unconditioned stimulus (US). However, it is possible that acquisition is primarily determined not by the number of trials but rather by quantities that often correlate with the number of trials, such as cumulative intertrial interval (ITI) and the number of sessions. Four experiments examined whether the number of trials has an effect on acquisition of conditioned responding, once cumulative ITI and number of sessions are equated. Results of the experiments with rats and mice favor the hypothesis that over an eightfold range, variation in number of CS-US pairings has little effect. It is suggested that learning curves might more accurately be plotted across cumulative ITI or number of sessions, and not across number of trials. Results pose a challenge to trial-centered accounts of conditioning, as demonstrated by simulations of the Rescorla-Wagner model, a simplified version of Wagner's standard operating procedure model (SOP), and Stout & Miller's sometimes competing retrieval model (SOCR). A time-centered account, rate estimation theory (RET), predicts the main finding but has trouble with other aspects of the learning process more easily accommodated by trial-centered models. PMID- 18426304 TI - The influence of CS-US interval on several different indices of learning in appetitive conditioning. AB - Four experiments examined the effects of varying the conditioned stimulus unconditioned stimulus (CS-US) interval (and US density) on learning in an appetitive magazine approach task with rats. Learning was assessed with conditioned response (CR) measures, as well as measures of sensory-specific stimulus-outcome associations (Pavlovian-instrumental transfer, potentiated feeding, and US devaluation). The results from these studies indicate that there exists an inverse relation between CS-US interval and magazine approach CRs, but that sensory-specific stimulus-outcome associations are established over a wide range of relatively long, but not short, CS-US intervals. These data suggest that simple CR measures provide different information about what is learned than measures of the specific stimulus-outcome association, and that time is a more critical variable for the former than latter component of learning. PMID- 18426305 TI - Contextual control of inhibition with reinforcement: adaptation and timing mechanisms. AB - Four experiments with rats studied the effects of switching the context after Pavlovian conditioning. In three conditioned suppression experiments, a large number of conditioning trials created "inhibition with reinforcement" (IWR), in which fear of the conditional stimulus (CS) reached a maximum and then declined despite continued CS-unconditional stimulus pairings. When IWR occurred, a context switch augmented fear of the CS; IWR and augmentation were highly correlated. Neither IWR nor augmentation resulted from inhibition of delay (IOD): In conditioned suppression, IWR and augmentation occurred without IOD (Experiment 3), and in appetitive conditioning (Experiment 4), IOD occurred without IWR or augmentation. IWR may occur in conditioned suppression because the animal adapts to fear of the CS in a context-specific manner. The authors discuss several implications. PMID- 18426306 TI - Timing during inhibitory conditioning. AB - Analyses of second-by-second conditioned responding into a food receptacle by hungry rats (Rattus norvegicus) found that inhibition varies across the duration of a conditioned stimulus (CS) in a manner consistent with initial training. Variations in the arrival time of the unconditioned stimulus (US) supported temporally specific suppression of responding (Experiment 1). Summation and retardation tests (Experiments 2 and 3, respectively) revealed that points of greatest inhibition coincided with US omission at the time normally specified by the excitor. Our data provide a clear demonstration of fine-grained changes in the time course of inhibitory conditioning for the first time. PMID- 18426307 TI - Excitatory and inhibitory learning with absent stimuli. AB - Three experiments showed that 2 associatively activated stimulus representations may engage in excitatory or inhibitory learning, depending on their temporal relationship. Experiment 1a suggested that simultaneously activated stimulus representations show evidence of inhibitory learning in an acquisition test. Experiment 1b showed similar evidence of inhibition in a summation test. Experiment 2 found that activation of 2 stimulus representations in a serial compound resulted in excitatory learning between the antecedent and the subsequent (forward) and inhibitory learning between the subsequent and the antecedent (backward). The results show the dynamic influence of temporal contiguity on mediated learning. PMID- 18426308 TI - Unblocking in Pavlovian fear conditioning. AB - Six experiments used rats to study blocking and unblocking of fear learning. An excitatory stimulus (A) blocked fear learning to a neutral stimulus (B). Unblocking of B occurred if the AB compound signaled an increase in unconditioned stimulus (US) intensity or number. Assessments of associative change during blocking showed that more was learned about B than A. Such assessments during unblocking revealed that more was learned about B than A following an increase in US intensity but not US number. These US manipulations had no differential effects on single-cue learning. The results show that variations in US intensity or number produce unblocking of fear learning, but for each there is a different profile of associative change and a potentially different mechanism. PMID- 18426309 TI - Memory without awareness: pigeons do not show metamemory in delayed matching to sample. AB - Metamemory, the ability to report on memory strength, is clearly established in rhesus macaques (Macaca mulatta) by converging evidence from several paradigms. In contrast, A. Inman and S. J. Shettleworth (1999) found no conclusive evidence of metamemory in pigeons. The authors studied pigeons further in 3 paradigms, with multiple tests of metamemory in each. Pigeons encountered a safe alternative to a matching-to-sample test either before (Experiment 2) or concurrently with (Experiment 3) the matching test. Choices of the safe option did not vary consistently with matching accuracy or change in trials with omitted samples in the way predicted for an animal with metamemory. In Experiment 4, confidence ratings following completion of the matching test also did not vary consistently as predicted by metamemory. PMID- 18426310 TI - Flavor preferences produced by backward pairing with wheel running. AB - In 3 experiments rats given 8 sessions of preexposure to wheel running acquired a preference for a flavor that was given immediately after each of 4 subsequent sessions of wheel running. Such flavor preference was less likely when rats were given the same conditioning procedure but without preexposure to wheels (Experiment 1) or when access to flavor was delayed by 30 min following a wheel session (Experiment 2). When rats were given a flavor before each wheel session, the resulting conditioned aversion was greater in rats that had no prior exposure to wheel running (Experiment 3). These results show that whether an aversion or preference for a flavor is produced by wheel running depends on an interaction between prior wheel experience and the sequence of events. PMID- 18426311 TI - Context conditional flavor preferences in the rat based on fructose and maltodextrin reinforcers. AB - In three experiments, rats were exposed to a flavor preference procedure in which flavor A was paired with the reinforcer and flavor B presented alone in Context 1, while in Context 2 flavor A was presented alone and flavor B with the reinforcer. With fructose as the reinforcer both two- and one-bottle training procedures produced a context-dependent preference (Experiments 1 and 2). With maltodextrin as the reinforcer two-bottle training produced a context-dependent preference (Experiment 1). Following one-bottle training with maltodextrin reinforcement rats demonstrated a context-dependent preference when the conditioned stimulus (CS)- was presented with a dilute solution of the reinforcer during training (Experiment 3B) but not when the CS- was presented alone (Experiments 2 and 3A). The pattern of results with maltodextrin reinforcement suggests that there was competition between the cue flavors and the taste of the maltodextrin as predictors of the postingestive consequences of the maltodextrin reinforcer. The fact that rats were able to display context-dependent flavor preferences is consistent with the idea that learned flavor preferences rely on the sort of cue-consequence associations that underpin other forms of conditioning which produce accurate performance on biconditional tasks. The differences between fructose- and maltodextrin-based preferences are discussed in terms of configural and elemental learning processes. PMID- 18426312 TI - Comparing elemental and configural associative theories in human causal learning: a case for attention. AB - In two causal learning experiments with human participants, the authors compared various associative theories that assumed either elemental (unique cue, modified unique cue, replaced elements model, and Harris' model) or configural processing of stimuli (Pearce's theory and a modification of it). The authors used modified patterning problems initially suggested by Redhead and Pearce (1995). Predictions for all theories were generated by computer simulations. Both configural theories and the unique cue approach failed to account for the observations. The replaced elements model was able to account for part of the data, but only if the replacement parameters could vary across discrimination problems. The Harris model and the modified unique cue approach, assuming that the salience of stimuli decreases with an increasing number of stimuli in a compound, successfully accounted for all of our data. This success implies that attentional factors should be explicitly taken into account in associative learning theory. PMID- 18426313 TI - Frequency of BCR-ABL fusion transcripts in Iranian patients with chronic myeloid leukemia. AB - BACKGROUND: A specific chromosomal abnormality, the Philadelphia chromosome, is present in 90 - 95% of patients with chronic myeloid leukemia. The aberration results from a reciprocal translocation of chromosomes 9 and 22, creating a BCR ABL fusion gene. There are two major forms of the BCR-ABL fusion gene, involving ABL exon 2, but including different exons of BCR gene. The transcript b2a2 or b3a2 codes for a p210 protein. Other fusion gene leads to the expression of an e1a2 transcript, which codes for a p190 protein. Other less common fusion genes are b3a3 or b2a3 (p203) and e19a2 (p230). The incidence of one or other rearrangement in chronic myeloid leukemia patients varies in different reports. In general, fusion transcripts are determined individually, a process which is labor- intensive in order to detect all major fusion transcripts. The objective of this study was to set up a multiplex RT-PCR assay for detection and to determine the frequency of different fusion genes in 75 Iranian patients with chronic myeloid leukemia. METHODS: Peripheral blood samples were analyzed by multiplex RT-PCR from 75 adult Iranian chronic myeloid leukemia patients to detect different types of BCR-ABL transcripts of the t(9;22). RESULTS: All patients examined were positive for some type of BCR/ABL rearrangement. The majority of the patients (83%) expressed one of the p210BCR-ABL transcripts (b3a2, 62% and b2a2, 20%), while the remaining showed one of the transcripts of b3a3, b2a3, e1a2 or co-expression of b3a2 and b2a2. The rate of co-expression of the b3a2 and b2a2 was 5%. CONCLUSION: In contrast to other reports, we did not see any co-expression of p210/p190. Co-expression may be due to alternative splicing or to phenotypic variation, with clinical course different from classic chronic myeloid leukemia. PMID- 18426314 TI - Linkage and association of DRD2 gene TaqI polymorphism with schizophrenia in an Iranian population. AB - BACKGROUND: D2 dopamine receptor gene has been reported to be one of the most relevant candidate genes in schizophrenia. In this study, we investigated the association between TaqIA and TaqIB dopamine D2 receptor polymorphisms and psychopathology of schizophrenia. METHODS: The study subjects were 38 acutely exacerbated schizophrenic patients who were all Iranian descent. The control population consisted of 63 healthy individuals with almost the same age as patients and were also of Iranian decent. The TaqIA and TaqIB genotypes, the A1 and A2 alleles, and the B1 and B2 were determined by restriction fragment length polymorphism of the amplified DNA fragments by polymerase chain reaction . RESULTS: For each polymorphism (A or B) the patients were categorized according to their genotype into three groups; i.e. the patients with alleles A1/A1, A1/A2, A2/A2; B1/B1, B1/B2, and B2/B2. No significant association was found between Taq1A or Taq1B gene polymorphisms and schizophrenia in patients compared to the controls. When study subjects were stratified according to their gender, the distribution of the A1/A1 genotype did was significantly different in both men and women (patients vs. controls). CONCLUSION: Our findings show that there is no genetic association between Taq1A and Taq1B gene polymorphisms and schizophrenia. Further clinical studies should be conducted to confirm and further evaluate these findings. PMID- 18426315 TI - Factors influencing editors' decision on acceptance or rejection of manuscripts: the authors' perspective. AB - BACKGROUND: There are few reports in the scientific literature on the factors taken into account by editors in deciding to accept or reject a scientific paper. The purpose of the present study was to investigate the effects of different factors on the journal editors' decisions on whether to accept or reject the manuscripts submitted to their journals. METHODS: We randomly selected the participants from the authors of original articles and case reports published in six medical journals, and sent them a questionnaire by e-mail. We analyzed the scores they gave to each of the 17 items of the questionnaire. RESULTS: One hundred and nineteen of the authors responded to our survey. The scores given by the respondents were analyzed comparing authors of developing and developed countries. Also, the results from authors of high-impact journals were compared with those with a low-impact factor. Multidimensional scaling was used to categorize the items based on their average scores. Highest scores were given to items addressing the quality of study performance, those addressing manuscript writing, and to the role of statistical significance of the results in the probability of studies getting published. CONCLUSION: Authors still believe in the existence of publication bias. They estimate its role to be comparable with the role of the quality of study performance and reporting. Our study also proves the presence of developing country bias, from the authors' perspective. PMID- 18426316 TI - Disability and depression in patients with chronic pain: pain or pain-related beliefs? AB - BACKGROUND: Physical disability and depression in patients with chronic pain have been shown to be associated with pain intensity and pain self-efficacy beliefs. However, little is known about whether pain self-efficacy beliefs can predict depression and physical disability when this relationship is controlled for pain intensity and other related demographic variables The aim of the current study was to replicate and extend previous research on the relationship between pain related beliefs, depression, and disability by examining these relationships in a heterogeneous sample of Iranian patients with chronic pain. METHODS: A group of 430 patients with chronic pain participated in the study and completed questionnaires on demographic variables, pain intensity, pain self-efficacy beliefs, physical disability, and depression. RESULTS: Correlation analyses revealed that patients with higher education were less depressed and less physically disabled. Younger patients were more physically disabled. Pain intensity and pain self-efficacy beliefs were significantly related to physical disability and depression. In hierarchical multiple regression analyses, after controlling for patients' background variables and pain intensity, pain self efficacy beliefs accounted for significant variance in depression and physical disability over and above the effect of demographic variables and pain intensity. Patients with higher pain self-efficacy, compared to those with lower self efficacy, were less depressed and less physically disabled. CONCLUSION: Pain self efficacy was more strongly related to depression and physical disability than pain intensity and demographic variables. The findings of the present study suggest the importance of targeting pain self-efficacy beliefs for modification in treatment of patients with chronic pain. PMID- 18426317 TI - Is Bristow-Latarjet operation effective for every recurrent anterior shoulder dislocation? AB - BACKGROUND: The treatment of recurrent anterior shoulder dislocation in patients who failed a supervised rehabilitation program is operative stabilization. Anatomical repair addressing the underlying pathology is the preferred method. We hypothesize that Bristow-Latarjet procedure is effective in all types of traumatic recurrent anterior shoulder dislocations, although in cases with Bankart lesion, Bankart operation is certainly preferred. METHODS: Thirty-five shoulders on which a Bristow-Latarjet operation had been performed on account of traumatic recurrent anterior shoulder instability were followed up for an average of 24.6 months (range: 18 - 51). The clinical outcome was measured according to Walch-Duplay Rating Sheet for Anterior Instability of the Shoulder at the latest follow-up. RESULTS: The clinical outcome was excellent in 11 (317%) patients and good in 24 (69%) patients. Two patients had redislocation that were treated nonoperatively and remained symptom-free at the latest follow-up. Radial nerve palsy occurred in one patient that spontaneously recovered after nine weeks. CONCLUSION: Although the treatment of anterior shoulder instability in patients with Bankart lesion and intact capsular material (without excessive laxity) is certainly Bankart operation, we claim that in almost all types of anterior shoulder instability, especially in patients with large Hill-Sachs lesions, glenoid bone loss, or excessive capsular laxity, the Bristow-Latarjet operation is associated with good or excellent results and can make the patient satisfied. PMID- 18426318 TI - Association of body mass index and abdominal obesity with marital status in adults. AB - BACKGROUND: Obesity continues to be an important public health problem worldwide. The objective of this study was to determine the association of body mass index and abdominal obesity with current marital status among the adult population of Iran. METHODS: A nation-wide cross-sectional survey was conducted from December 2004 through February 2005.The subjects were selected by stratified probability cluster sampling through household family members in Iran. Weight, height, waist circumference, and marital status of 89,404 men and women aged 15 - 65 (mean: 39.2) years were recorded. Four classes of body mass index, i.e., <18.5, 18.5 - 24.9, 25 - 29.9, and > or =30 kg/cm2, and three marital status, i.e., currently-, formerly-, and never-married were used. Abdominal obesity was defined as waist circumference > or =102 cm in men and > or =88 cm in women. RESULTS: The prevalence of overweight was twofold higher in married men (OR: 2.24; 95% CI: 2.08 - 2.41) and women (OR: 2.36; 95% CI: 2.20 - 2.53) than never-married men and women, even when age, educational level, leisure time physical activity, smoking habits, and place of residence were controlled. The multivariate OR of obesity was increased about threefold in married men (2.82; 95% CI: 2.51 - 3.18) and women (3.64; 95% CI: 3.31 - 3.99). The prevalence of abdominal obesity was twofold higher among married men (2.02; 95% CI: 1.79 - 2.29) and about threefold higher among married women (2.87; 95% CI: 2.69 - 3.06). CONCLUSION: The marital status appears to influence the likelihood of developing overweight, obesity, and abdominal obesity in both men and women in Iran. PMID- 18426319 TI - Combined fascia and mesh closure of large incisional hernias. AB - BACKGROUND: Incisional hernias are common and recurrence after repair has been reported in up to 44% of patients. Large incisional hernias of the abdominal wall represent substantial defect of supportive tissues. METHODS: Twenty-nine patients with large incisional hernias underwent surgery from January 2003 through December 2005. Herein, we presented our experience in closure of large incisional hernias using a technique in which we combine a fascia with a prosthetic repair. The variables recorded were classified as patient-related (gender, age, obesity, cough, constipation, diabetes mellitus, glucocorticosteroid therapy, smoking habit, and abdominal surgical history) and operation-related factors (size of defect, recurrence, wound infection, hematoma, and duration of hospital stay). RESULTS: The repair was performed for 25 midline hernias and four large incisional hernias in the right subcostal region. Four patients were females and 25 were males with a mean age of 52 (range: 30 - 77) years. The mean size of fascial defect was 12.7x4.5 cm. The mean time of operation was three hours and 18 minutes. The mean hospital stay was six (range: four to ten) days. Two patients had recurrence during the follow-up period. The mean follow-up period was 16 (range: eight to 26) months. CONCLUSION: The combined three-layer fascia and mesh repair can be successfully used for large incisional hernias. PMID- 18426320 TI - Effect of dextromethorphan on antinociception and tolerance induced by swim stress in the formalin test. AB - BACKGROUND: In the present study, the effect of dextromethorphan on antinociception and tolerance induced by water swim-stress in the formalin test was investigated. METHODS: Swim-stress at 8 masculineC induces antinociception in both phases of the formalin test. Intraperitoneal administration of dextromethorphan (60 mg/kg) also induces antinociception in the second phase of the formalin test. The lower doses of dextromethorphan (1.25, 2.5, and 5 mg/kg) which did not induce antinociception alone, but did so in combination with swim- stress (40 second), showed antinociceptive effect in both phases of the test. Exposure to water swimming stress with a period of 20 sec, once daily for three days, altered swim- stress- induced antinociception in the formalin test, when tested on the fourth day. RESULTS: In these animals, exposure to either water swimming stress alone or water swimming stress in combination with dextromethorphan showed potentiation of antinociception induced by swim- stress up to 20 second and decreased the response induced by 40 and 60 second swim- stress, indicating a tolerance induction. Dextromethorphan (20 mg/kg) did not alter the changes induced by three days exposure to swim- stress. CONCLUSION: The results may indicate a possible involvement of n-methyl-d-aspartate receptor mechanism in the antinociception but not tolerance induced by swim- stress at 8 masculineC. PMID- 18426321 TI - Smoking and gut. AB - Smoking has not only enormous deleterious effects on cardiovascular, cerebral, and bronchial organs but also profoundly alters the function of all parts of the gastrointestinal tract through various mechanisms. Except the sole curiously observed benefit of smoking on the course of ulcerative colitis, it increases the prevalence of the common gastrointestinal diseases namely gastroesophageal acid reflux, peptic ulcer, and Crohn's diseases. It also increases the incidence of cancer of oral cavity, esophagus, stomach, pancreas, and liver mostly in a dose response relationship and worsens the prognosis of colon cancer. The cessation of smoking is associated with the reduced incidence of cancer in the reported organs, but its effect on the regression of benign disease is not generally studied. The physicians must be aware not only of the harmful effect of smoking on the cardiovascular and bronchial systems, but also about the detrimental consequences of life-long smoking on the gastrointestinal tract and the increase of its benign and malignant diseases. PMID- 18426322 TI - A new horizon in primary prevention of cardiovascular disease, can we prevent heart attack by "heart polypill"? PMID- 18426323 TI - Pharyngocutaneous fistula after laryngectomy: incidence, predisposing factors, and outcome. AB - Pharyngocutaneous fistula is a common and troublesome postoperative complication after total laryngectomy. The objective of this report was to determine the incidence, predisposing factors, and outcome of postlaryngectomy pharyngocutaneous fistula in patients operated on in our department and to describe the management of the complication.The medical records of 146 consecutive patients who underwent laryngeal surgery for squamous cell carcinoma of the larynx between 1990 and 2005 were assessed. All patients had similar preoperative/postoperative care. We studied a number of factors that could influence fistula formation such as age, gender, smoking, systemic disease, preoperative radiotherapy, previous tracheotomy, site of tumor, surgical procedure, positive surgical margins, type of closure (T vs. vertical), concurrent neck dissection, suture material, clinical stage, histologic grade, and experience of surgeon (consultant vs. resident). A pharyngocutaneous fistula was observed in 13% (19/146) of the patients within a mean time of 9.6 days from surgery. Spontaneous closure with local wound care was noted in 17 (89%) patients whereas a surgical closure was necessary in two. One patient required surgical closure by direct suture of the pharyngeal mucosa. Pectoralis major myocutaneous flap was used in another one. Our findings showed that fistula formation was significantly more common in patients who received previous radiotherapy or who had positive surgical resection margins or had a systemic disease. The mean healing time was 26 days. We concluded that pharyngocutaneous fistula remains a troublesome complication of the early postoperative period after total laryngectomy. There are many conflicting reports in the literature concerning the predisposing factors, but our data showed that the presence of systemic diseases, previous radiotherapy, and positive surgical margins can all be important predisposing factors, or at least underlying causes. Our experience confirmed that most fistulas can be successfully managed with conservative treatment. PMID- 18426324 TI - The impact of low serum triglyceride on LDL-cholesterol estimation. AB - Most clinical laboratories directly measure serum triglyceride, total cholesterol, and high- density lipoprotein cholesterol. They indirectly calculate low-density lipoprotein cholesterol value using the Friedewald equation. Although high serum triglyceride (>400 mg/dL or 4.52 mmol/L) devaluates low-density lipoprotein cholesterol calculation by using this formula, effects of low serum triglyceride (<100 mg/dL or 1.13 mmol/L) on its accuracy is less defined.Two hundred thirty serum samples were assayed during a one-year period. In 115 samples, the triglyceride level was below 100 mg/dL and in 115 samples from age- and sex-matched patients the triglyceride level was 150 - 350 mg/dL (1.69 - 3.95 mmol/L). In both groups total cholesterol was above 250 mg/dL (6.46 mmol/L). On each sample, total cholesterol, high-density lipoprotein cholesterol, and triglyceride were directly measured in duplicate and low-density lipoprotein cholesterol measured directly and calculated with Friedewald equation as well. Statistical analysis showed that when triglyceride is <100 mg/dL, calculated low- density lipoprotein cholesterol is significantly overestimated (average :12.17 mg/dL or 0.31 mmol/L), where as when triglyceride is between 150 and 300 mg/dL no significant difference between calculated and measured low-density lipoprotein cholesterol is observed. In patients with low serum triglyceride and undesirably high total cholesterol levels, Friedewald equation may overestimate low-density lipoprotein cholesterol concentration and it should be either directly assayed or be calculated by a modified Friedewald equation. Using linear regression modeling, we propose a modified equation. PMID- 18426325 TI - The prevalence of coronary risk factors in Iranian lor migrating tribe. AB - Cardiovascular diseases are the major causes of death in Iran. The aim of this study was to determine the prevalence of conventional risk factors for coronary artery disease in Lor migrating tribes in southern part of Iran. Two hundred six persons of Lor migrating tribes in Mamasani, southern Iran (age range, 21 - 80 years) were randomly enrolled in the study. Their serum total cholesterol, triglyceride, high- density lipoprotein, low- density lipoprotein, and fasting blood sugar were determined. Of the participants, 79.9% were smokers. The prevalence of hypertension was 37.4% (mostly grade 1), 3.6% had diabetes mellitus, and 14.7% had impaired fasting glucose. A cholesterol level of more than 240 mg/dL was found in 7.1% of the participants, 27.8% had a low-density lipoprotein level of more than 130 mg/dL, 38.9% had a high-density lipoprotein level of less than 40 mg/dL, and 25.2% had a triglyceride level of more than 200 mg/dL. Body mass index greater than 25 was found in 34.8% of the participants, and 57.1% had intermediate and high risk for coronary events. The prevalence of conventional risk factors for coronary artery disease in Lor migrating tribes in comparison with Tehran urban population was very high. PMID- 18426326 TI - Localized myxedema of the toe: a rare presentation of graves' dermopathy. AB - Infiltrative dermopathy is an uncommon manifestation of Graves' disease. The most frequent location of infiltration is the lower extremities, especially the pretibial areas and on the dorsum of the foot. Rarely the hand, elbow, arm, and forearm are affected. We report a 66-year-old man with Graves' disease of nine years duration who presented with dermopathy and nonpitting edema of the toe with no involvement of the pretibial portion since four years ago. Radiologic studies of the foot were normal, without subperiosteal bone formation or reaction. Skin biopsy was consistent with pretibial myxedema. This presentation of myxedema and limitation to a localized area for several years is very unusual. PMID- 18426327 TI - Vitamin D-dependent rickets type II: report of a novel mutation in the vitamin D receptor gene. AB - Hereditary vitamin D-resistant rickets type or vitamin D-dependent rickets type II is a genetically determined and rare autosomal recessive disorder, most often caused by mutations in the vitamin D receptor gene. It usually presents with rachitic changes not responsive to vitamin D treatment and the circulating levels of 1,25 (OH)2 vitamin D-3 are elevated, differentiating it from vitamin D dependent rickets type I. Alopecia capitis or alopecia totalis is seen in some families with vitamin D-dependent rickets type II. This is usually associated with a more severe phenotype. In this report, we present the clinical findings on a family which exhibited the typical clinical features of hereditary vitamin D resistant rickets in two siblings. In addition, molecular analysis of the vitamin D receptor gene was performed by sequencing all coding exons. The cardinal findings in the index patient were alopecia totalis, renal tubular acidosis, mild generalized aminoaciduria, refractory rickets, high alkaline phosphatase, and hyperparathyroidism. Other routine biochemical tests were within normal limits, but 1+ glycine was detected in his urine. Skin biopsy results were compatible with alopecia areata. A previous child with similar phenotype was reported to be deceased at the age of 32 months. Mutation analysis of the vitamin D receptor gene by direct sequencing analysis of all coding exons showed a homozygous c.122GA(p.Cys41Tyr) variant in exon 2 with several arguments pointing to a pathogenic effect. We should be aware of this very rare disease whenever we see a patient with refractory rickets and alopecia. PMID- 18426328 TI - Neonatal diabetes mellitus due to pancreatic agenesis. AB - Neonatal diabetes mellitus, which is defined as hyperglycemia presenting within the first six weeks of life, is a rare disorder. It may result in transient or permanent disease. Pancreatic agenesis is a rare cause of neonatal diabetes.We report a neonate who was small for gestational age and presented with diabetes mellitus and signs of malabsorption because of pancreatic agenesis. PMID- 18426330 TI - Squamous cell carcinoma of the tongue in a 13-year-old boy. AB - Squamous cell carcinoma of the tongue is uncommon among children and teenagers. Most commonly, squamous cell carcinoma of the head and neck presents during the fifth and sixth decades of life in patients with a long history of tobacco and alcohol use. The rarity of this lesion in young patients implies that clinicians might not include it in the differential diagnoses. In general, carcinoma of the oral cavity in young people is reported to be particularly aggressive and carry a poor prognosis. A case of carcinoma of the tongue in a 13-year-old boy is presented in this report. PMID- 18426329 TI - Concomitant hypoparathyroidism, sensorineural deafness, and renal agenesis: a case of Barakat syndrome. AB - Barakat syndrome, also known as hypoparathyroidism, sensorineural deafness, and renal dysplasia syndrome is an extremely rare congenital disorder. Different etiologies are described for the syndrome but the definite pathophysiology remains unclear. Hereby, we present a case of Barakat syndrome who was diagnosed on the basis of clinical and molecular data. PMID- 18426331 TI - Photoclinic. PMID- 18426332 TI - A brief history of the discovery of the circulation of blood in the human body. AB - The present article describes briefly the development of the theories regarding the circulation of blood in humans, from the time of Galen (second century C.E.) to the work of William Harvey (17th century C.E.).We shall summarize the views of Galen together with those of two prominent Iranian physicians of the Middle Ages (Razi and Ahwazi known in the West as Rhazes and Haly Abbas respectively) as well as that of Ibn-Nafis from Damascus (the discoverer of the pulmonary circulation) and the Spanish physician and cleric Michael Servetus and finally the definitive work of William Harvey, the English physician who described the mechanism of both the systemic and pulmonary circulation of blood in the human body. PMID- 18426334 TI - Health behaviors and weight status among urban and rural children. AB - INTRODUCTION: Pediatric overweight is currently reaching epidemic proportions but little information exists on differences in weight related behaviors between urban and rural children. OBJECTIVE: To assess health behaviors and weight status among urban and rural school-age children. METHODS: Fifth-grade children at two urban and two rural schools were invited to participate in an assessment study of their health behaviors and weight status. A total of 138 children (mean age = 10 years; % female = 54.6) chose to participate. RESULTS: Children in rural and urban areas consumed equivalent calories per day and calories from fat, but rural children ate more junk food and urban children were more likely to skip breakfast. Urban children engaged in more metabolic equivalent tasks and had slightly higher total sedentary activity than rural children. The BMI percentile was equivalent across rural and urban children but rural children were more often overweight and urban children were more often at risk for overweight. CONCLUSIONS: Although some variables were equivalent across urban and rural children, results indicate some key health behavior differences between groups. Results should be interpreted with caution as the sample size was small and there were demographic differences between urban and rural samples. PMID- 18426335 TI - Molecular epidemiology and divergence of HIV type 1 protease codon 35 inserted strains among treatment-naive patients in Hong Kong. AB - This study reported the prevalence and pattern of viral replication-associated HIV-1 protease codon 35 amino acid insertions among treatment-naive patients in Hong Kong. The transmission and divergence date of these inserted strains was also investigated. The pol gene of 264 local HIV-1 isolates was sequenced and phylogenetic analysis was performed. The transmission history of protease codon 35-inserted HIV-1 strains in Hong Kong was estimated by the Bayesian coalescent method. This insertion was detected in 12 (4.55%) among 264 treatment-naive subtype B HIV-1 patients in Hong Kong, which was 20-times higher than the prevalence in the western countries. Among these strains, eight carried a glutamic acid (GAA) insertion (E35E_E), two carried an aspartic acid (GAC) insertion (E35E_D), and two carried a glycine (GGA) insertion (E35E_G). E35E_D and E35E_E insertions were the first to be reported. All the 12 inserted sequences clustered in the same lineage of the phylogenetic tree, indicating the possibility of transmission of this insertion. Epidemiological investigation revealed the major route of infection for this inserted strain in Hong Kong was associated mainly among homosexual Chinese males. The evolutionary rate of these inserted strains was similar to other subtype B HIV-1 strains. Through coalescent based analysis, the divergence date of the protease codon 35-inserted strains in Hong Kong was 1995. Our findings demonstrate the epidemic pathways of viral fitness-related HIV-1 protease codon 35-inserted isolates in Hong Kong. The effect of these novel insertions on viral fitness and drug susceptibility requires further investigation. PMID- 18426336 TI - Male genital tract compartmentalization of human immunodeficiency virus type 1 (HIV). AB - We present phylogenetic evidence supporting viral compartmentalization between the blood (peripheral blood mononuclear cells or plasma) and multiple genitourinary sites in HIV-infected men. Four of the five subjects evaluated demonstrated compartmentalization of viral sequences between urogenital tract specimens (tissue or fluid) and at least one blood category. HIV sequence migration from blood to urogenital tract was detected in four of five men, with migration from urogenital tract to blood in the fifth, and cross migration between both compartments noted in one man. These observations add 5 additional cases to the 27 total reported cases in which male urogenital tract compartmentalization has been studied, investigate surgical samples/specimens that have not been evaluated previously, and provide further evidence for restricted flow of HIV between the blood and the genital tract. As such, our study findings are important for understanding the long-term response to antiretroviral therapy, the design of vaccines, and the sexual transmission of HIV. PMID- 18426337 TI - Analysis of the immune responses in chimpanzees infected with HIV type 1 isolates. AB - The mechanisms of resistance to AIDS development in HIV-1-infected chimpanzees have remained elusive. Unique among chimpanzees naturally or experimentally infected with HIV, several animals of the Yerkes cohort have progressed to clinical AIDS with selection of isolates showing increased pathogenicity for chimpanzees. We compared progressors vs. nonprogressors among the HIV-infected chimpanzees that made up this cohort, eight of which have been infected with HIV 1 for over 14 years. The additional two progressors were infected de novo with chimpanzee-pathogenic HIV, rapidly leading to a progressor status. Nonprogressors were characterized by normal CD4(+) T cell counts and the absence of detectable viremia. In contrast, progressor chimpanzees had relatively high plasma viral loads associated with a dramatic loss of CD4(+) T cells. The analysis of immune responses showed a similar amplitude and breadth of ELISPOT T cell responses in both groups. HIV-specific proliferative responses were, however, absent in the progressor animals, which also exhibited increased levels of immune activation characterized by elevated levels of the circulating chemokines IP-10 and MCP-1. Of interest was the conservation of potent NK cell activity in all animals, potentially contributing to the extended symptom-free survival of progressor animals. Modest anti-HIV antibody titers were detectable in the nonprogressor group, but these antibodies exhibited good neutralizing activity. In progressors, however, two sets of data were noted: in animals that gradually selected for pathogenic isolates, or that were superinfected, very high neutralizing antibody titers were observed, although none to the pathogenic HIV. In contrast, two animals infected de novo with chimpanzee pathogenic HIV failed to mount an extensive humoral response and both failed to develop neutralizing antibodies to the virus. Taken together, pathogenic HIV infection in chimpanzees is associated with rapid loss of CD4(+) T cells and proliferative responses as well as higher levels of immune activation. PMID- 18426338 TI - A splenic marginal zone-like peripheral blood CD27+B220- B cell population is preferentially depleted in HIV type 1-infected individuals. AB - Peripheral blood CD27(+) B cells are reduced in HIV-1-infected individuals. In healthy individuals, the human peripheral blood CD27(+) B cell pool consists of two subsets defined by the expression, or lack thereof, of the CD45 isoform B220. We investigated the presence of circulating B220(+) and B220(-) memory B cells in HIV(+) individuals and found that the reduction in CD27(+) memory B cells occurs primarily among CD27(+)B220(-) B cells. Studies conducted using healthy controls indicate that CD27(+)B220(-) B cells have a splenic marginal zone like the immunophenotype IgM(hi)IgD(lo)CD21(+)CD23(-), express TLR9, and proliferate and secrete IgG and IgM in response to B cell-specific ODN. CD27(+)B220(+) B cells have the immunophenotype IgM(lo)IgD(hi)CD21(+)CD23(+), express activation-induced cytidine deaminase, and proliferate in response to SAC but do not secrete immunoglobulin. The AICD expression, along with CD86 expression, by CD27(+)B220(+) suggests these cells are of germinal center origin. The preferential depletion of CD27(+)B220(-) B cells mirrors alterations in spleen morphology and resident B cell populations due to HIV infection reported by other investigators and may play an important role in the defective B cell immunity against T-independent pathogens such as pneumococcus observed in HIV-1-infected individuals. PMID- 18426340 TI - iPS cells: a more critical review. AB - Over the past 20 months, reports claiming the generation of induced pluripotent stem (iPS) cells with characteristics identical to those of embryonic stem (ES) cells from nonembryonic tissue have captured great attention in both the scientific community and the general public. In the light of the continuing controversy over the use of ES cells, these reports have profound ramifications. This review calls into question the validity of many claims made in these reports -claims that have led to the rapid and premature acceptance of using iPS cells as a viable alternative to using normal stem cells for regenerative therapy. How convincing is the evidence supporting the various claims made for the iPS cells? Are there other more plausible explanations for the same observations? What are these iPS cells? Are they really safe for therapeutic use? Should the iPS technique be considered, in the absence of any direct evidence for induction and reprogramming, as a realistic alternative for somatic cell nuclear transfer (SCNT) to generate ES-like cells? This review attempts to encourage reflections on and offer alternative views for key aspects of iPS cells and studies. PMID- 18426341 TI - When competition does not matter: grassland diversity and community composition. AB - We examined whether the intense root competition in a rough fescue grassland plant community in central Alberta, Canada, was important in structuring plant species diversity or community composition. We measured competition intensity across gradients of species richness, evenness, and community composition, using pairs of naturally occurring plants of 12 species. One plant in each pair was isolated from neighbors to measure competition; community structure and environmental conditions were also measured at each pair. We used structural equation modeling to examine how competition influenced community structure. Competition intensity was unrelated to species richness and community composition, but increased competition intensity was associated with a slight decline in evenness. Size-symmetric root competition was probably unimportant in structuring this plant community because there are no feedback mechanisms through which size-symmetric competition can magnify small initial differences and eventually lead to competitive exclusion. In plant communities with little shoot competition, competition and community structure should be unlinked regardless of competition intensity. In more productive systems, we propose that interactions between root and shoot competition may indirectly structure communities by altering the overall asymmetry of competition. PMID- 18426339 TI - Derivation of neural stem cells from mesenchymal stemcells: evidence for a bipotential stem cell population. AB - Neural stem cell (NSC) transplantation has been proposed as a future therapy for neurodegenerative disorders. However, NSC transplantation will be hampered by the limited number of brain donors and the toxicity of immunosuppressive regimens that might be needed with allogeneic transplantation. These limitations may be avoided if NSCs can be generated from clinically accessible sources, such as bone marrow (BM) and peripheral blood samples, that are suitable for autologous transplantation. We report here that NSCs can be generated from human BM-derived mesenchymal stem cells (MSCs). When cultured in NSC culture conditions, 8% of MSCs were able to generate neurospheres. These MSC-derived neurospheres expressed characteristic NSC antigens, such as nestin and musashi-1, and were capable of self-renewal and multilineage differentiation into neurons, astrocytes, and oligodendrocytes. Furthermore, when these MSC-derived neurospheres were cocultured with primary astrocytes, they differentiate into neurons that possess both dendritic and axonal processes, form synapses, and are able to fire tetrodotoxin-sensitive action potentials. When these MSC-derived NSCs were switched back to MSC culture conditions, a small fraction of NSCs (averaging 4 5%) adhered to the culture flasks, proliferated, and displayed the morphology of MSCs. Those adherent cells expressed the characteristic MSC antigens and regained the ability to differentiate into multiple mesodermal lineages. Data presented in this study suggest that MSCs contain a small fraction (averaging 4-5%) of a bipotential stem cell population that is able to generate either MSCs or NSCs depending on the culture conditions. PMID- 18426342 TI - Counting coup. PMID- 18426343 TI - Coming of age. PMID- 18426344 TI - Higher body mass index predicts need for insulin but not hyperglycemia, nosocomial infection, or death in critically ill surgical patients. AB - BACKGROUND: Strict glycemic control in critically ill patients has been an important advance in surgical critical care, as hyperglycemia is associated with a higher likelihood of death, complications, and nosocomial infections. Insulin resistance is particularly common in obese patients, but the impact of body mass index (BMI) on insulin requirements, ability to achieve euglycemia, and infectious outcomes in critically ill surgical patients has not been studied. We hypothesized that obese patients would not incur a higher likelihood of infection if euglycemia was maintained. METHODS: Admissions to the surgical intensive care unit (ICU) from October 1, 2004, to October 31, 2006, were identified. Necessary data were available for 946 patients. The main predictor variable was BMI, which was analyzed as both a continuous and a five-level categorical variable. Data on insulin requirements as well as glycemic control were captured. The main outcome variable was the occurrence of at least one nosocomial infection. Additional outcomes were dysfunction of at least one organ system at any time during surgical ICU admission, quantified using the Multiple Organ Dysfunction Score, as well as the ICU length of stay and death. All statistical analyses were performing using SPSS version 11 for Macintosh. RESULTS: Both the need for insulin infusion (p = 0.0001) and the mean insulin units/day among patients receiving infusions (p = 0.03) increased significantly with increasing BMI. However, periods of euglycemia were similar among BMI groups. A total of 152 patients (16.1%) incurred at least one nosocomial infection, for a total of 169 infections. The majority (n = 107; 63.3%) were ventilator-associated pneumonias. Neither infection (p = 0.99), organ dysfunction (p = 0.14), ICU length of stay (p = 0.22), nor mortality rate (p = 0.09) differed significantly by BMI group. The need for an insulin infusion was associated significantly with nosocomial infection (p = 0.0001). Additional predictors of infection were a higher Acute Physiology and Chronic Health Evaluation (APACHE) III score (p < 0.0001), age adjusted APACHE III score (p < 0.0001), and emergency admission (0.001). After controlling for the need for an insulin infusion, BMI was not associated with infection. CONCLUSIONS: Increasing BMI was associated significantly with insulin resistance. Despite insulin resistance, however, obese patients did not incur longer periods of hyperglycemia. Outcomes that have been associated consistently with glycemic control, such as nosocomial infection and mortality rate, did not differ according to BMI. These data suggest that BMI is not associated with infection during critical illness, and that this absence of an association may be influenced at least partially by the ability to maintain similar glycemic control in obese and non-obese patients. PMID- 18426345 TI - Effect of body mass index and ertapenem versus cefotetan prophylaxis on surgical site infection in elective colorectal surgery. AB - BACKGROUND: The effectiveness of prophylactic antibiotics in the prevention of surgical site infection (SSI) after elective colorectal surgery is dependent on many factors, including the body mass index (BMI) of the patient. In this study, the association of BMI and type of antibiotic prophylaxis with SSI was evaluated in patients undergoing elective colorectal surgery. METHOD: A post-hoc analysis was performed using data obtained from a multicenter randomized, double-blind study of 1,002 patients undergoing elective colorectal surgery who received prophylactic administration of ertapenem (1 g) or cefotetan (2 g). Among 650 evaluable patients, the effect of BMI and type of antibiotic prophylaxis on SSI rates was assessed four weeks after surgery. Mechanical bowel preparation was standardized, and no patient received oral antibiotics; intravenous antibiotics were not repeated during or after surgery. RESULTS: The majority of patients had a BMI between 18.5 and 39.9 kg/m2. Regardless of the type of prophylaxis, SSI rates were significantly higher in patients with a BMI > or = 30 kg/m2 than in those with a BMI < 30 kg/m2. However, failure, defined as SSI, was significantly less common after ertapenem than after cefotetan prophylaxis at both BMI < 30 kg/m2 (12.7% vs. 26.4%, respectively; difference -13.7; 95% confidence interval [CI] -21.0, -6.5) and BMI > or = 30 kg/m2 (26.7% vs. 41.9%, respectively; difference -15.3; 95% CI -28.2, -2.0). The most prevalent type of SSI was superficial incisional infection, which was more common with both treatments in patients with a BMI > or = 30 kg/m2; however, the incidence of superficial SSI was lower after ertapenem than cefotetan prophylaxis. CONCLUSION: In patients undergoing elective colorectal surgery, the incidence of SSI, specifically superficial incisional SSI, was higher in patients with a BMI > or = 30 kg/m2, regardless of the prophylactic antibiotic given. Ertapenem prophylaxis was more effective than cefotetan in the prevention of SSI at any BMI. PMID- 18426346 TI - Outbreak of resistant Pseudomonas aeruginosa infections during a quarterly cycling antibiotic regimen. AB - BACKGROUND: Antibiotic cycling or rotation of antimicrobial agent classes has been proposed to combat antimicrobial resistance. METHODS: A prospective cohort study was conducted in a medical intensive care unit (ICU) of a university hospital between December 1, 2000, and September 30, 2002, as part of a three center trial under the aegis of the U.S. Centers for Disease Control and Prevention. Patients admitted to the medical ICU for > 48 h were enrolled, and demographic and microbiological data were collected until discharge or death. Baseline data were collected for four months (12/1/00 to 3/31/01) and compared with data collected after institution of a quarterly cycling regimen (cycle order: Cefepime, ciprofloxacin, piperacillin-tazobactam, imipenem-cilastatin) for the empiric treatment of gram-negative infections (4/01/01 to 9/30/02). RESULTS: Of 1,074 consecutive admissions, 301 were enrolled, 59 during baseline and 242 during the cycling periods. An outbreak of multi-drug resistant Pseudomonas aeruginosa followed cycle 2 (cefepime), coinciding with cycles 3 and 4 (ciprofloxacin and piperacillin-tazobactam) (80.0 and 73.7 vs. 37.3 isolates/100 patients enrolled for cycles 3/4 and baseline, respectively; p = 0.04). Acinetobacter spp. were isolated less frequently during the cycling periods (15.3 vs. 1.2 isolates/100 patients for baseline and cycling periods, respectively; p > or = 0.01). The crude hospital mortality rate was similar (24/59 [41%] baseline vs. 73/242 [30%] cycling; p = 0.16) between periods. However, the percentage of patients admitted to the medical ICU who subsequently acquired an infection followed by in-hospital death was higher at baseline than during cycling: 15/59 (25.4%) vs. 33/242 (13.6%)(p = 0.04). CONCLUSIONS: In this study, the cycling strategy was not definitively associated with beneficial changes in unit epidemiology and in fact may have contributed to an outbreak of multi-drug resistant P. aeruginosa. PMID- 18426347 TI - Pseudomonas aeruginosa susceptible only to colistin in intensive care unit patients. AB - BACKGROUND: Gram-negative bacilli, including multi-drug-resistant (MDR) Pseudomonas aeruginosa, are responsible for severe intensive care unit (ICU) acquired infections, mainly pneumonia and bacteremia. The aim of this study was to determine the incidence of MDR strains of Pseudomonas in patients undergoing cardiac surgery, to elucidate the effectiveness of treating these patients with colistin, and to assess the safety of the drug. METHODS: A prospective study was conducted among 1,452 patients who underwent surgery for a variety of cardiac lesions over a one-year period, and who spent a portion of the recovery period in the surgical ICU. Their case histories were analyzed to identify infectious complications. Diagnosis of infection was based on clinical data, and the pathogen was tested with respect to its susceptibility to colistin (polymyxin E). The clinical response to the antibiotic was evaluated. RESULTS: Over the 12-month period, among 115 infected patients, 15 were affected by strains of P. aeruginosa. In 10 patients, this pathogen proved resistant to all potentially active antibiotics except colistin. All of the affected patients were being ventilated mechanically, and eight of them presented with ventilator-associated pneumonia (VAP), whereas one patient suffered a deep incisional surgical site infection and bacteremia and the remaining patient had a superficial infection of a lower-extremity vein graft donor site. The MDR pathogen was introduced to the hospital by three patients transferred from three institutions. All patients were treated with intravenous colistin. In cases of VAP, aerosolized colistin was added. Deterioration of renal function occurred in three patients (30%), all of whom had a history of renal insufficiency. Cure or clinical improvement was observed in seven patients (70%), whereas four patients, including one who improved initially, developed sepsis and died with multiple organ dysfunction syndrome (mortality rate 40%). CONCLUSIONS: The increasing prevalence of MDR P. aeruginosa in ICU patients has rekindled interest in polymyxins, which had been abandoned because of toxic side effects. Colistin retained significant in vitro activity against this virulent organism, had an acceptable safety profile, and should be considered as a treatment option in critically ill patients with infection caused by MDR gram-negative bacilli. Aerosolized colistin may merit further consideration as a therapeutic intervention for patients with refractory pulmonary infections. PMID- 18426348 TI - Systemic administration of enamel matrix derivative to lipopolysaccharide challenged pigs: effects on the inflammatory response. AB - BACKGROUND: Periodontitis is the primary clinical indication for enamel matrix derivative (EMD). Recent investigations, showing that EMD inhibits the production of tumor necrosis factor-alpha (TNF-alpha) when added to human whole blood, indicate a novel role for EMD as a modulator of systemic inflammation. In the present study, we investigated the systemic effects of EMD in lipopolysaccharide (LPS)-challenged pigs. METHODS: In a preparatory study, seven pigs received a prophylactic EMD bolus injection (5 mg/kg), followed by a continuous infusion (50 mg/kg/min). Thirty minutes later, a continuous infusion of LPS (1.7 mcg/kg/h) was started. An additional 12 pigs were randomized into two groups. Six of these animals were given the same treatment, except that EMD was administered 30 min after LPS. The remainder served as controls. The groups were compared according to organ injury and function, hemodynamics, and systemic markers of inflammation. RESULTS: Prophylactic administration of EMD triggered transient hemodynamic instability in two of seven pigs. In the randomized pigs, no or only nonspecific changes were observed in biopsies from vital organs, independent of treatment. Enamel matrix derivative did not modify systemic TNF-alpha, interleukin (IL)-1 beta, or IL-6 concentrations. CONCLUSIONS: In the formulation and dosages used, EMD did not modulate the inflammatory response. No true allergic or immunotoxic reactions were seen. To be usable for systemic application, a new formulation should be developed, or the active part of the protein(s) should be identified and produced in a soluble form designed for infusion. The potential of EMD as a systemic immune modulator is still unsettled. PMID- 18426349 TI - Blood stream infections of abdominal origin in the intensive care unit: characteristics and determinants of death. AB - BACKGROUND: Blood stream infections (BSI) of abdominal origin are associated with a high mortality rate. We hypothesized that both patient and microbiological factors determine death in critically ill patients who develop such infections. METHODS: Ninety-six consecutive patients who developed BSI of abdominal origin in an 11-year period (1992-2002) in the intensive care unit (ICU) of the Ghent University Hospital were studied. Patient data were retrieved from a prospective registry of BSI. Demographics, disease severity, source of the BSI, incidence of organ failure, and outcome were recorded. Microbiological data were retrieved from the patient file and the hospital laboratory. RESULTS: Secondary peritonitis and intra-abdominal abscesses were the source of the BSI in the majority of patients. The majority of the organisms involved were gram-negative, with Escherichia coli isolated most frequently. Twenty-one patients (22%) had polymicrobial BSI, and in 39 patients, at least one of the micro-organisms was antibiotic resistant (41%). The mortality rate in the whole patient group was 62.5% (60/96), which was significantly higher than the Acute Physiology and Chronic Health Evaluation (APACHE) II-based expected mortality rate (p < 0.001). Patients who died were older, had a tendency to have a higher APACHE II score on admission, and were more likely to suffer from acute renal failure and cardiovascular failure during their ICU stay. Logistic regression analysis revealed that the following factors were independently associated with death: Age (odds ratio [OR] 1.09; 95% confidence interval [CI] 1.04, 1.14; p < 0.001) (per year increase) and the occurrence of acute renal failure (OR 4.18; 95% CI 1.22, 14.31; p = 0.023). CONCLUSIONS: The mortality rate of ICU patients who develop BSI of intra-abdominal origin is high. Gram-negative micro-organisms were isolated most frequently, and 41% of all organisms were antibiotic-resistant. Two patient-related factors (greater age and the development of acute renal failure) were associated independently with a higher mortality rate. PMID- 18426350 TI - Incidence of postoperative infections in non-immunocompromised pediatric patients with low absolute neutrophil counts preoperatively. AB - PURPOSE: To discover the incidence of postoperative surgical site infections in non-immunocompromised pediatric patients with an absolute neutrophil count (ANC) < or = 1,000 preoperatively. METHODS: Office and hospital charts of consecutive patients with preoperative ANC < or = 1,000 undergoing elective surgery over a three-year period were reviewed for evidence of postoperative surgical site infection. RESULTS: Six of 200 patients with preoperative ANCs < or = 1,000 developed a postoperative surgical site infection, an infection rate of 3.0%. One of 35 patients with preoperative ANCs < or = 500 developed a surgical site infection. The mean ANC of the study population was 800 (range 200 to 1,000); the mean ANC of the group with postoperative infection likewise was 800 (range 500 to 1,000). CONCLUSIONS: There was a postoperative surgical site infection rate of 3.0% in non-immunocompromised children who had a preoperative ANC < or = 1,000, similar to the overall surgical site infection rate in children. Cancellation of elective procedures in non-immunocompromised children with ANC < or = 1,000 is not warranted on the grounds of concern about postoperative infection. PMID- 18426352 TI - Treatment of metronidazole-refractory Clostridium difficile enteritis with vancomycin. AB - BACKGROUND: Clostridium difficile infection of the colon is a common and well described clinical entity. Clostridium difficile enteritis of the small bowel is believed to be less common and has been described sparsely in the literature. METHODS: Case report and literature review. RESULTS: We describe a patient who had undergone total proctocolectomy with ileal pouch-anal anastomosis who was treated with broad-spectrum antibiotics and contracted C. difficile refractory to metronidazole. The enteritis resolved quickly after initiation of combined oral vancomycin and metronidazole. A literature review found that eight of the fifteen previously reported cases of C. difficile-associated small-bowel enteritis resulted in death. CONCLUSIONS: It is important for physicians who treat acolonic patients to be aware of C. difficile enteritis of the small bowel so that it can be suspected, diagnosed, and treated. PMID- 18426351 TI - Statin (cerivastatin) protects mice against sepsis-related death via reduced proinflammatory cytokines and enhanced bacterial clearance. AB - BACKGROUND: Death attributable to septic shock syndrome depends highly on the inflammatory response and cytokine production. Inhibition of inflammation is one of the many pleiotropic effects of statins. The aim of this study was to test the hypothesis that statins have a role in altering the host response to bacterial infections and thus would prove beneficial in the prevention of microbial sepsis. METHODS: Male C57BL/6 and C3H/HeN mice received cerivastatin (4 mg/kg) or phosphate-buffered saline (PBS) intraperitoneally (i.p.), 24 and 1 h before and 24, 48, and 72 h after bacterial challenges. Sepsis was induced by i.p. injection of lipopolysaccharide (LPS) (45 mg/kg), Staphylococcus aureus (5 x 10(7) colony forming units [CFU]/mouse), or Salmonella typhimurium (2.5 x 10(6) CFU/mouse). RESULTS: Administration of cerivastatin improved significantly the survival rate of mice challenged with LPS (31% vs. 19% in the PBS group; p = 0.001), S. aureus (56% vs. 20% in PBS group; p = 0.01), or S. typhimurium (48% vs. 10% in PBS group; p = 0.03). Significantly reduced release of the pro-inflammatory cytokines tumor necrosis factor (TNF)-alpha and interleukin (IL)-6 was evident in cerivastatin-treated mice after LPS challenge. Cerivastatin-treated mice showed insignificant reductions in serum TNF-alpha and IL-6 concentrations after bacterial challenge. However, significantly accelerated bacterial clearance was demonstrated in cerivastatin-treated mice 24 h after S. typhimurium infection and 48 h after S. aureus infection. CONCLUSIONS: Cerivastatin protects mice against LPS- and live bacteria-induced death, an effect associated with cerivastatin attenuated pro-inflammatory cytokine production and enhanced bacterial clearance. Hence, application of statins in the clinical setting may prove beneficial in prevention of LPS or bacterial infection-related sepsis. PMID- 18426353 TI - Acinetobacter baumannii mediastinitis after cardiopulmonary bypass: case report and literature review. AB - BACKGROUND: Mediastinitis resulting from surgical site infection may occur in 1% of patients undergoing median sternotomy. METHODS: Case report and review of the pertinent English-language literature. RESULTS: We report a case of mediastinitis caused by Acinetobacter baumannii, in a patient with multiple comorbidities who underwent cardiopulmonary bypass. Successful treatment consisted of surgical debridement, reconstruction, and ampicillin-sulbactam. CONCLUSIONS: Acinetobacter baumannii should be recognized as a potential causative agent of severe postoperative mediastinitis. PMID- 18426354 TI - Local anesthetics as antimicrobial agents: a review. AB - BACKGROUND: Since the introduction of cocaine in 1884, local anesthetics have been used as a mainstay of pain management. However, numerous studies over the past several decades have elucidated the supplemental role of local anesthetics as antimicrobial agents. In addition to their anesthetic properties, medications such as bupivacaine and lidocaine have been shown to exhibit bacteriostatic, bactericidal, fungistatic, and fungicidal properties against a wide spectrum of microorganisms. METHODS: A comprehensive literature search was conducted using MEDLINE 1950-present for in vitro and in vivo studies pertaining to the antimicrobial activity of various local anesthetics on a broad range of bacterial and fungal pathogens. Studies testing the effect on microbial growth inhibition of local anesthetics alone and in combination with other agents, such as preservatives and other medications, as well as the effect of conditions such as concentration and temperature, were included for review. Outcome measures included colony counts, area-under-the-curve and time-kill curve calculations, minimum inhibitory concentrations, and post-antibiotic effect. RESULTS: Evidence suggests that local anesthetics as a class possess inherent antimicrobial properties against a wide spectrum of human pathogens. Multiple local anesthetics at concentrations typically used in the clinical setting (e.g., bupivacaine 0.125%-0.75%; lidocaine 1%-3%) inhibit the growth of numerous bacteria and fungi under various conditions. Different local anesthetics showed various degrees of antimicrobial capacity; bupivacaine and lidocaine, for example, inhibit growth to a significantly greater extent than does ropivacaine. Greater concentrations, longer exposure, and higher temperature each correlate with a proportional increase in microbial growth inhibition. Addition of other agents to the anesthetic solutions, such as preservatives, opioids, or intravenous anesthetics such as propofol, modify the antimicrobial activity via either synergistic or antagonistic action. Limited studies attribute the mechanism of action of antimicrobial activity of local anesthetics to a disruption of microbial cell membrane permeability, leading to leakage of cellular components and subsequent cell lysis. CONCLUSIONS: Local anesthetics not only serve as agents for pain control, but possess antimicrobial activity as well. In such a capacity, local anesthetics can be considered as an adjunct to traditional antimicrobial use in the clinical or laboratory setting. Additionally, caution should be exercised when administering local anesthetics prior to diagnostic procedures in which culture specimens are to be obtained, as the antimicrobial activity of the local anesthetic could lead to false-negative results or suboptimal culture yields. PMID- 18426355 TI - Is vasopressin increasing or decreasing mortality in patients with septic shock? PMID- 18426356 TI - The image of surgical infection: hidradenitis suppurativa and Ancylostoma braziliense. PMID- 18426362 TI - A novel homozygous missense mutation of the dual oxidase 2 (DUOX2) gene in an adult patient with large goiter. AB - OBJECTIVE: To describe the first adult case of large goiter associated with a novel R1110Q mutation in the dual oxidase 2 (DUOX2) gene. She was initially euthyroid, and developed hypothyroidism later in her forties. DUOX2 is an essential enzyme in iodine organification of thyroid hormone biosynthesis. Only infant cases of congenital hypothyroidism due to mutations of the DUOX2 gene have been reported. Biallelic mutation of DUOX2 is thought to lead to total iodine organification defect. PATIENTS AND MEASUREMENT: This 57-year-old woman became first aware of goiter around the age of 20 years. Since the goiter had enlarged gradually, she consulted us at the age of 32 years. Goiter was soft, and thyroid function was normal. Antithyroid antibodies were negative. Both physical and mental development was normal. Three of her nine siblings and her mother had large goiters. At the age of 44 years, thyroid function demonstrated subclinical hypothyroidism. She started to take levo-thyroxine at a dose of 100 mug/day to reduce goiter. At the age of 56 years, goiter size remained the same. The perchlorate discharge rate was 72.8%, suggesting partial iodine organification defect. Thus, thyroid peroxidase (TPO) gene and DUOX2 gene were analyzed. RESULTS: There was no mutation in the TPO gene, but a novel homozygous mutation (R1110Q) in the DUOX2 gene was identified. The same heterozygous mutation was detected in her two sons and two grandchildren. This mutation was not detected in 104 control alleles and was located at a site differing from any other reported mutations in the DUOX2 gene. CONCLUSIONS: This homozygous missense mutation can be associated with thyroid dysfunction and goiter formation of an enlarged thyroid gland. PMID- 18426363 TI - Review on the occasion of a decade of recombinant human TSH: prospects and novel uses. AB - The introduction of recombinant human thyroid-stimulating hormone (rhTSH) almost a decade ago represents a remarkable achievement in the history of clinical thyroidology. rhTSH now contributes substantially to the diagnostic approach to thyroid cancer, offering a reliable and safe alternative to thyroid hormone withdrawal by avoiding the morbidity of hypothyroidism. Several recent studies have also demonstrated the efficacy of radioiodine ablation of thyroid remnants after preparation with rhTSH. Moreover, the use of rhTSH in this context is associated with a lower whole body exposure to radiation compared to thyroid hormone withdrawal. Although not approved officially, rhTSH-assisted treatment of locoregional or distant metastatic disease may be the treatment of choice for patients in whom hypothyroidism may be relatively contraindicated, such as the very young and the aged, although additional efficacy studies are necessary. The compound has also been shown to be useful in the treatment of nontoxic multinodular goiter, especially when the radioiodine uptake is low. Finally, rhTSH may prove to be useful in studying the functional reserve of thyroid in the aging process, as well as the putative role of the TSH receptor in extrathyroidal tissue, such as lymphocytes, osteocytes, and adipocytes. PMID- 18426364 TI - Unsatisfactory consequences from fine-needle aspiration biopsy of thyroid nodules. PMID- 18426365 TI - Stool cultures and antimicrobial prescriptions related to infectious diarrhea. AB - Stool cultures can be important in guiding antimicrobial therapy for diarrhea. From among 11.64 million person-years of Tennessee Medicaid enrollment data collected from 1995 through 2004, 315,828 diarrheal episodes were identified. Stool cultures were performed for only 15,820 episodes (5.0%). Antimicrobials were prescribed for 32,949 episodes (10.4%), 89.4% of which were not accompanied by a stool culture. White race and urban residence were associated with higher rates of stool culture. Frequent use of antimicrobials for diarrhea without stool culture may indicate inappropriate antimicrobial use and has critical implications for public health. PMID- 18426366 TI - Multidrug-resistant and extensively drug-resistant tuberculosis: the National Institute of Allergy and Infectious Diseases Research agenda and recommendations for priority research. PMID- 18426367 TI - Prevalence of and risk factors for anal human papillomavirus infection in heterosexual men. AB - In US men, the incidence of anal cancer, the primary cause of which is human papillomavirus (HPV) infection, has increased almost 3-fold in 3 decades; however, little is known about the epidemiology of anal HPV infection, especially in heterosexual men. In 2 US cities, behavioral data and anal biological specimens were collected from 253 men who acknowledged having engaged in sexual intercourse with a woman during the preceding year. On the basis of DNA analysis, overall prevalence of anal HPV infection was found to be 24.8% in 222 men who acknowledged having had no prior sexual intercourse with men. Of the men with anal HPV infection, 33.3% had an oncogenic HPV type. Risk factors independently associated with anal HPV were lifetime number of female sex partners and frequency of sex with females during the preceding month. These results suggest that anal HPV infection may be common in heterosexual men. PMID- 18426370 TI - Changing patterns in HIV reverse transcriptase resistance mutations after availability of tenofovir. AB - Assessment of 1177 human immunodeficiency virus (HIV) resistance genotypes at an HIV/AIDS clinic showed a decrease in the incidence of the K65R mutation, from 15.2% of isolates during the period 2002-2004 to 2.7% of isolates during the period 2005-2006 (P < .001), despite elevated and stable rates of tenofovir use. A reduction in the rate of coadministration of didanosine (from 41.6% of patients in 2004 to 0.8% of patients in 2006; P < .001) largely explained this observation. PMID- 18426372 TI - The emerging crisis of drug-resistant tuberculosis in South Africa: lessons from New York City. AB - Multidrug-resistant tuberculosis and extensively drug-resistant tuberculosis have emerged as important infections in South Africa among patients infected with human immunodeficiency virus (HIV). In the face of this new epidemic, South Africa must rededicate itself to the task of tuberculosis control and treatment with a rapid, multifaceted approach. Priorities include expansion of second-line treatment capacity, investment in clinical laboratories, a system to ensure supervised treatment for all patients, and enhancement of infection control procedures. In New York City, where drug-resistant tuberculosis emerged 2 decades ago--also in the context of a large HIV-infected population and an underfunded public health infrastructure--similar steps were successful in leading to the rapid decrease in rates of drug resistance among tuberculosis isolates. With refinements based on local resource constraints, urgent measures could potentially arrest the alarming increase in multidrug-resistant and extensively drug-resistant tuberculosis cases in South Africa. Unlike many countries in sub Saharan Africa, South Africa has the capacity to mount a rapid and large-scale response before drug-resistant tuberculosis envelops a much larger and far poorer region. PMID- 18426373 TI - An outbreak of human external ophthalmomyiasis due to Oestrus ovis in southern Afghanistan. AB - Oestrus ovis is the most common cause of human ophthalmomyiasis, and infection is often misdiagnosed as acute conjunctivitis. Although it typically occurs in shepherds and farmers, O. ovis ophthalmomyiasis has also been reported in urban areas. We report the first case study of O. ovis infection from Afghanistan. PMID- 18426374 TI - Unsuspected, disseminated coccidioidomycosis without maternofetal morbidity diagnosed by placental examination: case report and review of the literature. AB - Historically, untreated disseminated coccidioidomycosis during pregnancy was thought to be associated with 100% maternal fatality and 50% fetal mortality and was the leading cause of maternal deaths in areas of endemicity. As recently as 1995, therapeutic abortions and early deliveries were advocated in certain contexts. This report describes an unrecognized case of disseminated coccidioidomycosis diagnosed at the time of placental examination in a woman who completed her pregnancy without significant maternofetal complications. This case suggests that abortion and early delivery may not be necessary, because the possibility of an uncomplicated pregnancy exists. It is likely that other similar cases exist but remain underreported or underdiagnosed because of the mild, nondescript nature of the illness and low clinical suspicion. Although this mother and infant had good clinical outcomes, thorough travel histories and consideration of the associated travel-related diseases are important because of the possibility of serious, potentially avoidable clinical consequences. PMID- 18426375 TI - Identifying barriers to hypertension care: implications for quality improvement initiatives. AB - The role of clinical inertia in the treatment of patients with hypertension was assessed by evaluating health care providers' knowledge, attitudes, and clinical practices regarding hypertension management. A cross-sectional survey was conducted at the Forsyth Medical Group in North Carolina. Participants were physicians (N = 18, 10 sites) and support staff (N = 20, 12 sites), who were surveyed in 2006. Physician and support staff questionnaires consisted of 29 and 15 items, respectively, and were administered by trained interviewers. Though most physicians (94%) cited familiarity with the Seventh Report of the Joint National Committee on Prevention, Detection, Evaluation, and Treatment of High Blood Pressure (JNC-7) guidelines and affirmed that hypertension management guidelines are relevant to their patients, no physicians interviewed routinely document patient hypertension management plans. Although 1 in 3 physicians cited the inability to devote enough time to patients for the discussion of hypertension management, physicians predominantly cited patient- and support staff- related factors as most important to patients not attaining blood pressure (BP) goal. Patient lifestyle modification (89%), education (67%), and medication compliance (56%) were cited as the most important reasons for uncontrolled BP. Only one-third of physicians believe that clinical staff always obtain accurate BP measurements, and 61% believe that resistant hypertension is a reflection of inaccurate BP measurement. Many support staff claimed to be rushed when measuring patient BP, and 65% recommended BP competency training. Contradictions were evident between provider knowledge of hypertension management standards and how this knowledge is applied in clinical practice. Standardized collection of BP is critical to measuring clinical improvement in hypertension. Results are being utilized to develop clinical improvement initiatives including staff education and competency training. PMID- 18426376 TI - Regression to the mean: a limited issue in disease management programs for chronic obstructive pulmonary disease. AB - Our objective was to test for evidence of regression to the mean in chronic obstructive pulmonary disease (COPD)-related health care utilization in a Colorado Medicaid population that met the criteria for, but were not participating in, a COPD disease management (DM) program. National Jewish Medical and Research Center had enrolled individuals who (1) had a diagnosis of COPD for at least 1 year and (2) were active participants in Colorado Medicaid's 1-year DM program called breatheWise; the present study sought a comparator group for that population. In order to test for evidence of regression to the mean (ie, high utilization from the recruitment period reducing without active intervention) in this case management model, we conducted a case-controlled analysis of total spending for a comparator population that would have met the inclusion criteria for the DM program. The present study assessed health care utilization for fiscal years 2002 and 2003 in terms of total rates of emergency room (ER) visits and hospitalizations for all causes in the comparator group of COPD patients. In addition, total costs related to both ER visits and hospitalizations were compiled. In total, 354 individuals met the inclusion criteria and were identified as the comparator group. ER visits and hospitalizations were consistent for 2002 and 2003. ER visits totaled 314 and 315 in 2002 and 2003, respectively, indicating a 0.3% increase that was not significant. Hospitalizations decreased from 0.53 admissions per patient in 2002 to 0.48 in 2003-a 9.4% reduction that was not significant. With comparable rates of ER visits and hospitalizations, total costs for health care utilization remained virtually unchanged between 2002 and 2003. There is minimal evidence of regression to the mean over 2 consecutive years in the Colorado Medicaid patients with moderate to severe COPD. PMID- 18426377 TI - Increased adherence to cardiac standards of care during participation in cardiac disease management programs. AB - Adherence to cardiovascular disease standards of care is critically important for minimizing the risk of mortality and morbidity for individuals with coronary heart disease (CHD) and heart failure (HF). The purpose of this study was to assess the ability of cardiac disease management (DM) programs to assist members with their adherence to evidence-based medicine for cardiovascular diseases. A total of 20,202 members with CHD and/or HF were evaluated 12 months prior to the start of DM programs and during their first 12 months of participation in the programs. Members were assessed for their adherence to appropriate cardiac medications. In addition, low-density lipoprotein (LDL) testing rates and clinical control of LDL values (defined as <100 mg/dL) were measured. The association between LDL control and use of lipid-lowering statins also was assessed. During participation in the cardiac programs, members achieved significant improvement in their adherence to angiotensin-converting enzyme inhibitors/angiotensin receptor blockers, and beta-blockers (P < 0.0001). The cardiac population also achieved a significant increase in LDL testing rates and statin use (P < 0.0001). More members attained appropriate LDL control in year 1 compared to baseline (36% relative increase), and this improvement was associated with a 40% relative increase in statin use. In summary, participation in these cardiac DM programs assisted members to improve their adherence to cardiac medications and standards of care guidelines. Such improvements in cardiovascular disease care are likely associated with improved quality of life and reduced risk for mortality. PMID- 18426371 TI - Long-term effects of highly active antiretroviral therapy on CD4+ cell evolution among children and adolescents infected with HIV: 5 years and counting. AB - BACKGROUND: Lower percentages of CD4(+) T lymphocytes are associated with adverse clinical outcomes among children and adolescents infected with human immunodeficiency virus (HIV). CD4(+) lymphocyte percentage generally increases with receipt of highly active antiretroviral therapy (HAART), but long-term follow-up is required to assess whether these increases in CD4(+) cell percentage are maintained and whether they lead to normal CD4(+) cell percentages in children with severe immunosuppression. METHODS: The study population included 1236 children and adolescents perinatally infected with HIV who were enrolled in a US-based multicenter prospective cohort study (Pediatric AIDS Clinical Trials Group 219/219C) and who were not receiving HAART at study initiation. We estimated the effects of HAART, HAART with protease inhibitors, and HAART with nonnucleoside reverse-transcriptase inhibitors on CD4(+) cell percentage, using marginal structural models to account for confounding by severity. RESULTS: Initiation of any type of HAART increased CD4(+) cell percentage by 2.34% (95% confidence interval, 1.35%-3.33%) in the first year, relative to noninitiation of HAART. The substantial increases in CD4(+) cell percentage observed after the first year of experience with these combination therapies were followed by relatively smaller increases that continued for 5 years after initiation. Although larger increases in CD4(+) cell percentage were observed among children with a greater degree of immunosuppression at baseline, the mean CD4(+) cell percentage after 5 years of HAART did not reach normal levels. CONCLUSIONS: Our study supports the initiation of HAART in children before severe immunosuppression occurs for long-term maintenance of normal CD4(+) cell percentages. This beneficial result must be weighed against the evidence of potential adverse events associated with the prolonged use of such therapy. PMID- 18426378 TI - Defining success in diabetes disease management: digging deeper in the data. AB - We evaluated the effectiveness of a diabetes life coach program designed to address the concerns of limited coordination and collaboration of care for chronically ill patients in the physician office. The program emphasized lipid, blood pressure, and glycemic control, using personal coaching, group classes, reminders, and customized feedback. The target population was all health plan members over age 18 with type 1 or 2 diabetes mellitus in 6 primary care practice sites in the Hampton Roads area of Virginia. Primary outcomes were 1 Health Plan Employer Data and Information Set measure (A1c poor control of >9% or no test), 3 American Diabetes Association (ADA) measures (A1c <7%, blood pressure of <130/80 mmHg, low-density lipoprotein cholesterol [LDL-C] of <100 mg/dL), 1 pharmacy measure (percentage of patients filling at least 1 insulin prescription), and 2 self-reported behavioral measures (percentage adherent to a meal plan and percentage adherent to an activity plan). We assessed overall program outcomes and differences between individual physician practices and evaluated outcomes separately for engaged compared with non-engaged program participants. Outcomes for 1117 participants were evaluated. Statistically significant improvement at P < 0.05 was noted in all 7 targeted measures compared with baseline. Participants who were engaged in the life coach program were 40% less likely to experience poor control of their A1c, 50% more likely to meet the ADA A1c goal of < 7%, 11% more likely to meet their blood pressure goal of <130/80 mmHg, and 7% more likely to meet their LDL-C goal of <100 mg/dL compared with those not engaged. Patients who became engaged in the program performed significantly better in the key diabetes indicators that ultimately lead to reductions in the complications of the disease over time. Our study contributes to the evidence that clinical multidisciplinary, collaborative models of care can influence and improve the management of diabetes. PMID- 18426379 TI - A qualitative study of the relationship between clinician attributes, organization, and patient characteristics on implementation of a disease management program. AB - The purpose of this study was to examine the challenges of integrating an asthma disease management (DM) program into a primary care setting from the perspective of primary care practitioners. A second goal was to examine whether barriers differed between urban-based and nonurban-based practices. Using a qualitative design, data were gathered using focus groups in primary care pediatric practices. A purposeful sample included an equal number of urban and nonurban practices. Participants represented all levels in the practice setting. Important themes that emerged from the data were coded and categorized. A total of 151 individuals, including physicians, advanced practice clinicians, registered nurses, other medical staff, and nonmedical staff participated in 16 focus groups that included 8 urban and 8 nonurban practices. Content analyses identified 4 primary factors influencing the implementation of a DM program in a primary care setting. They were related to providers, the organization, patients, and characteristics of the DM program. This study illustrates the complexity of the primary care environment and the challenge of changing practice in these settings. The results of this study identified areas in a primary care setting that influence the adoption of a DM program. These findings can assist in identifying effective strategies to change clinical behavior in primary care practices. PMID- 18426380 TI - Advances in PEGylation of important biotech molecules: delivery aspects. AB - BACKGROUND: Although various injected peptide and protein therapeutics have been developed successfully over the past 25 years, several pharmacokinetic and immunological challenges are still encountered that can limit the efficacy of both novel and established biotech molecules. OBJECTIVE AND METHOD: PEGylation is a popular technique to address such properties. PEGylated drugs exhibit prolonged half-life, higher stability, water solubility, lower immunogenicity and antigenicity, as well as potential for specific cell targeting. Although PEGylated drug conjugates have been on the market for many years, the technology has steadily developed in respect of site-specific chemistry, chain length, molecular weights and purity of conjugate. These developments have occurred in parallel to improvements in physicochemical methods of characterization. CONCLUSION: This review will discuss recent achievements in PEGylation processes with an emphasis on novel PEG-drugs constructs, the unrealized potential of PEGylation for non-injected routes of delivery, and also on PEGylated versions of polymeric nanoparticles, including dendrimers and liposomes. PMID- 18426381 TI - Polymers for viral gene delivery. AB - BACKGROUND: The development of viral vectors capable of providing efficient gene transfer in diseased tissues without causing any pathogenic effects is pivotal for overcoming the many challenges facing gene therapy. OBJECTIVE: Immune responses against viral vectors, inadequate gene expression and inefficient targeting to specific cells in vivo are some of the major problems limiting the clinical utility of viral gene therapy. METHODS: This review will focus on recent progress in strategic polymer-based modifications to improve the performance and biocompatibility of a variety of viral vectors. We will discuss the preclinical development of four approaches involving injectable polymers, polyelectrolytes, polymer microspheres and polymer-virus conjugates. RESULTS/CONCLUSION: Much progress has been made in creating 'hybrid' gene delivery vectors that combine the strengths of polymers and viruses. With further optimization, these hybrid vectors, which may be safer and more effective, are likely to succeed in clinical applications. PMID- 18426382 TI - Developments in polymeric devices for oral insulin delivery. AB - BACKGROUND: Development of improved oral insulin administration is necessary for the treatment of diabetes mellitus, to overcome the problem of daily subcutaneous injections. The vast amount of literature data on oral insulin delivery prompted us to cover this area in a review. OBJECTIVE: Insulin delivery using polymeric devices is discussed, with an ultimate aim of addressing the technological development in this area. METHODS: The development of oral delivery devices for insulin using hydrogels and micro/nanoparticles is discussed with reference to polymers. These efforts must be directed to increase the residence time of insulin near the intestinal absorptive cells. RESULTS/CONCLUSION: The published results on oral insulin delivery devices, particularly on inter-polymer complexes of the grafted copolymers, are discussed in greater depth. The use of absorption enhancers like cyclodextrins, bile salts and surfactants is covered. The state-of the-art technology and challenges in this area are discussed, with typical examples. PMID- 18426383 TI - Recent advances and perspectives on coated alginate microspheres for modified drug delivery. AB - BACKGROUND: Alginate microspheres represent a useful tool for modified drug delivery. Their preparation is quite easy and is usually based on the gelling properties of the polysaccharide in the presence of divalent ions; nevertheless, microparticles prepared only with calcium alginate show several problems, mainly related to the mechanical stability and to the release that, in most cases, is too fast. To overcome such inconveniences, polymer-coated alginate microspheres and/or appropriately interpenetrating polymer network (semi-IPNs and IPNs) structures formed with alginate and other macromolecules were developed. OBJECTIVE: This article reports a synthetic overview on the most recent searches carried out on coated alginate microspheres. METHODS: After a section focused on the microsphere preparation, this article is divided into several main topics related to the specific polymer that was used as a coating material to provide a rationale in reporting literature data. In the last section, the advantages and disadvantages of the various approaches are discussed and the authors' opinion on perspectives for further studies and novel applications of coated alginate microspheres are reported. CONCLUSION: Ca(2+)-alginate microparticles could experience a new era if scientists will increase their efforts in developing microparticles with smart properties. PMID- 18426384 TI - Application of drug delivery system to boron neutron capture therapy for cancer. AB - BACKGROUND: Tumor cell destruction in boron neutron capture therapy (BNCT) is due to the nuclear reaction between (10)B and thermal neutrons ((10)B + (1)n --> (7)Li + (4)He (alpha) + 2.31 MeV (93.7 %)/2.79 MeV (6.3 %)). The resulting lithium ions and alphaparticles are high linear energy transfer (LET) particles which give a high biological effect. Their short range in tissue (5 - 9 mum) restricts radiation damage to those cells in which boron atoms are located at the time of neutron irradiation. BNCT has been applied clinically for the treatment of malignant brain tumors, malignant melanoma, head and neck cancer and hepatoma. Sodium mercaptoundecahydro-dodecaborate (Na(2)(10)B(12)H(11)SH: BSH) and borono phenylalanine ((10)BPA) are currently being used in clinical treatments. These low molecule compounds are easily cleared from cancer cells and blood, so high accumulation and selective delivery of boron compounds into tumor tissues and cancer cells are most important to achieve effective BNCT and to avoid damage to adjacent healthy cells. OBJECTIVE: In order to achieve the selective delivery of boron atoms to cancer cells, a drug delivery system (DDS) is an attractive intelligent technology for targeting and controlled release of drugs. METHODS: We performed literature searches related to boron delivery systems in vitro and in vivo. RESULTS: We describe several DDS technologies for boron delivery to cancer tissues and cancer cells from the past to current status. We are convinced that it will be possible to use liposomes, monoclonal antibodies and WOW emulsions as boron delivery systems for BNCT clinically in accordance with the preparation of good commercial product (GCP) grade materials. PMID- 18426385 TI - Vector therapies for malignant glioma: shifting the clinical paradigm. AB - BACKGROUND: Malignant glioma represents one of the most aggressive and devastating forms of human cancer. At present, there exists no successful treatment for this disease. Gene therapy, or vector therapy, has emerged as a viable experimental treatment method for intracranial malignancies. OBJECTIVE: Vector therapy paradigms that have entered the clinical arena have shown adequate safety; however, the majority of the studies failed to observe significant clinical benefits. As such, researchers have refocused their efforts on developing novel vectors as well as new delivery methods to enhance the therapeutic effect of a particular vector. In this review, we discuss common vector therapy approaches used in clinical trials, their drawbacks and potential ways of overcoming these challenges. METHODS: We focus on the experimental evaluation of cell-based vector therapies and adenoviral and herpes simplex virus type 1 vectors in the treatment of malignant glioma. CONCLUSION: Vector therapy remains a promising treatment strategy for malignant glioma. Although significant questions remain to be answered, early clinical data suggest safety of this approach and future studies will likely address the efficacy of the proposed therapy. PMID- 18426386 TI - Aiming for the heart: targeted delivery of drugs to diseased cardiac tissue. AB - BACKGROUND: The development of a number of regenerative strategies in recent years for curing heart disease represents a paradigm shift away from conventional approaches which aim to manage heart disease. Effective administration of pharmaceutical agents targeted directly to the diseased tissue is the key to unlocking the potential of regenerative strategies, which could augment current conventional treatments. OBJECTIVE: The authors review recent advances in targeted drug delivery to diseased cardiac tissue. METHODS: Various therapeutic methodologies designed to selectively deliver pharmaceutical agents to diseased cardiac tissue are discussed in this review. CONCLUSION: Targeted delivery of survival and engraftment promoting factors to damaged cardiac tissue can be an important strategy, for example, in creating a suitable microenvironment encouraging the engraftment of stem cells. Further progress in this emerging field is contingent on the discovery of new biomarkers that are upregulated in damaged cardiac tissue and can be targeted for selective drug delivery. Once fully realized, breakthroughs in this field will have direct applications in the diagnosis and treatment of heart disease through more effective tissue-specific drug delivery and improved imaging modalities. PMID- 18426387 TI - Current and future testosterone delivery systems for treatment of the hypogonadal male. AB - BACKGROUND: Hypogonadism is manifest in all age groups, and a growing elderly population is requiring treatment for testosterone deficiency, presenting new safety challenges, as many of these individuals present with comorbidities and significant risk profiles. OBJECTIVE: To discuss testosterone replacement modalities, their advantages and disadvantages, and provide a discussion of safety issues. METHODS: We reviewed the literature regarding testosterone replacement therapy and have provided a summary of our most outstanding findings. CONCLUSION: Potential benefits of testosterone replacement therapy include increased lean body mass, heightened libido, increased bone density and elevation of mood. Some disadvantages are clearly defined, while others require further investigation. Patient and physician must cooperate to agree on an individual patient's most appropriate and tolerable route of administration. PMID- 18426388 TI - Trehalose 6-phosphate: a signal of sucrose status. AB - T6P (trehalose 6-phosphate), the precursor of trehalose, has come out of obscurity over 10 years to be appreciated as an important regulator of plant metabolism and development, quite possibly linking the two. This information has been gained from analysis of mutant and transgenic plants, which show strong, diverse and strategically important phenotypes. Plant genes that encode the trehalose pathway are numerous and highly regulated transcriptionally and post translationally, responding sensitively to the environment in a developmentally programmed and tissue-specific manner further suggestive of a vital function. Yet the precise role of T6P has not been clear. In an article published in the Biochemical Journal in 2006, John Lunn and colleagues addressed a major obstacle to understanding the function of T6P through development of a method capable of resolving femtomolar quantities of T6P from very small amounts of tissue. Using this technology, the authors showed large changes in T6P content that reflect tissue sucrose status. Overall, this elegant work makes an important contribution towards our understanding of the function of T6P in plants. PMID- 18426389 TI - Structural biology of plasmid partition: uncovering the molecular mechanisms of DNA segregation. AB - DNA segregation or partition is an essential process that ensures stable genome transmission. In prokaryotes, partition is best understood for plasmids, which serve as tractable model systems to study the mechanistic underpinnings of DNA segregation at a detailed atomic level owing to their simplicity. Specifically, plasmid partition requires only three elements: a centromere-like DNA site and two proteins: a motor protein, generally an ATPase, and a centromere-binding protein. In the first step of the partition process, multiple centromere-binding proteins bind co-operatively to the centromere, which typically consists of several tandem repeats, to form a higher-order nucleoprotein complex called the partition complex. The partition complex recruits the ATPase to form the segrosome and somehow activates the ATPase for DNA separation. Two major families of plasmid par systems have been delineated based on whether they utilize ATPase proteins with deviant Walker-type motifs or actin-like folds. In contrast, the centromere-binding proteins show little sequence homology even within a given family. Recent structural studies, however, have revealed that these centromere binding proteins appear to belong to one of two major structural groups: those that employ helix-turn-helix DNA-binding motifs or those with ribbon-helix-helix DNA-binding domains. The first structure of a higher-order partition complex was recently revealed by the structure of pSK41 centromere-binding protein, ParR, bound to its centromere site. This structure showed that multiple ParR ribbon helix-helix motifs bind symmetrically to the tandem centromere repeats to form a large superhelical structure with dimensions suitable for capture of the filaments formed by the actinlike ATPases. Surprisingly, recent data indicate that the deviant Walker ATPase proteins also form polymer-like structures, suggesting that, although the par families harbour what initially appeared to be structurally and functionally divergent proteins, they actually utilize similar mechanisms of DNA segregation. Thus, in the present review, the known Par protein and Par-protein complex structures are discussed with regard to their functions in DNA segregation in an attempt to begin to define, at a detailed atomic level, the molecular mechanisms involved in plasmid segregation. PMID- 18426390 TI - High membrane potential promotes alkenal-induced mitochondrial uncoupling and influences adenine nucleotide translocase conformation. AB - Mitochondria generate reactive oxygen species, whose downstream lipid peroxidation products, such as 4-hydroxynonenal, induce uncoupling of oxidative phosphorylation by increasing proton leak through mitochondrial inner membrane proteins such as the uncoupling proteins and adenine nucleotide translocase. Using mitochondria from rat liver, which lack uncoupling proteins, in the present study we show that energization (specifically, high membrane potential) is required for 4-hydroxynonenal to activate proton conductance mediated by adenine nucleotide translocase. Prolonging the time at high membrane potential promotes greater uncoupling. 4-Hydroxynonenal-induced uncoupling via adenine nucleotide translocase is prevented but not readily reversed by addition of carboxyatractylate, suggesting a permanent change (such as adduct formation) that renders the translocase leaky to protons. In contrast with the irreversibility of proton conductance, carboxyatractylate added after 4-hydroxynonenal still inhibits nucleotide translocation, implying that the proton conductance and nucleotide translocation pathways are different. We propose a model to relate adenine nucleotide translocase conformation to proton conductance in the presence or absence of 4-hydroxynonenal and/or carboxyatractylate. PMID- 18426391 TI - Cisplatin-mediated impairment of mitochondrial DNA metabolism inversely correlates with glutathione levels. AB - Cisplatin accumulates in mitochondria, which are a major target for this drug in cancer cells. Thus alterations in mitochondrial function have been implicated in cancer cell resistance to chemotherapeutic agents. Moreover, cisplatin toxic side effects seem to be associated with mitochondrial injury in vivo and in vitro. In order to clarify the potential effect of cisplatin in mtDNA (mitochondrial DNA) maintenance and expression, we have analysed rat liver mtDNA and mtRNA (mitochondrial RNA) synthesis as well as their stability under the influence of in vivo treatment or in vitro exposure to cisplatin. We show that cisplatin causes a direct and significant impairment of mtDNA and mtRNA synthesis and decreases steady-state levels of mtRNAs in isolated mitochondria. Furthermore, in vivo treatment of the animals with cisplatin exerts a protective effect from the impairment of mtRNA metabolism caused by in vitro exposure to the drug, by means of increased mitochondrial GSH levels after in vivo cisplatin treatment. PMID- 18426392 TI - Targeting human glutathione transferase A3-3 attenuates progesterone production in human steroidogenic cells. AB - hGSTA3-3 (human Alpha-class glutathione transferase 3-3) efficiently catalyses steroid Delta(5)-Delta(4) double-bond isomerization in vitro, using glutathione as a cofactor. This chemical transformation is an obligatory reaction in the biosynthesis of steroid hormones and follows the oxidation of 3beta hydroxysteroids catalysed by 3beta-HSD (3beta-hydroxysteroid dehydrogenase). The isomerization has commonly been ascribed to a supplementary function of 3beta HSD. The present study is the first to provide evidence that hGSTA3-3 contributes to this step in steroid hormone biosynthesis in complex cellular systems. First, we find glutathione-dependent Delta(5)-Delta(4) isomerase activity in whole-cell extracts prepared from human steroidogenic cells. Secondly, effective inhibitors of hGSTA3-3 dramatically decrease the conversion of Delta(5)-androstene-3,17 dione into Delta(4)-androstene-3,17-dione in cell lysates. Thirdly, we show that RNAi (RNA interference) targeting hGSTA3-3 expression decreases by 30% the forskolin-stimulated production of the steroid hormone progesterone in a human placental cell line. This effect is achieved at low concentrations of two small interfering RNAs directed against distinct regions of hGSTA3-3 mRNA, and is weaker in unstimulated cells, in which hGSTA3-3 expression is low. The results concordantly show that hGSTA3-3 makes a significant contribution to the double bond isomerization necessary for steroid hormone biosynthesis and thereby complements the indispensable 3beta-hydroxysteroid oxidoreductase activity of 3beta-HSD. The results indicate that the lower isomerase activity of 3beta-HSD is insufficient for maximal rate of cellular sex hormone production and identify hGSTA3-3 as a possible target for pharmaceutical intervention in steroid hormone dependent diseases. PMID- 18426393 TI - Proteomic analysis of polymeric salivary mucins: no evidence for MUC19 in human saliva. AB - MUC5B is the predominant polymeric mucin in human saliva [Thornton, Khan, Mehrotra, Howard, Veerman, Packer and Sheehan (1999) Glycobiology 9, 293-302], where it contributes to oral cavity hydration and protection. More recently, the gene for another putative polymeric mucin, MUC19, has been shown to be expressed in human salivary glands [Chen, Zhao, Kalaslavadi, Hamati, Nehrke, Le, Ann and Wu (2004) Am. J. Respir. Cell Mol. Biol. 30, 155-165]. However, to date, the MUC19 mucin has not been isolated from human saliva. Our aim was therefore to purify and characterize the MUC19 glycoprotein from human saliva. Saliva was solubilized in 4 M guanidinium chloride and the high-density mucins were purified by density gradient centrifugation. The presence of MUC19 was investigated using tandem MS of tryptic peptides derived from this mucin preparation. Using this approach, we found multiple MUC5B-derived tryptic peptides, but were unable to detect any putative MUC19 peptides. These results suggest that MUC19 is not a major component in human saliva. In contrast, using the same experimental approach, we identified Muc19 and Muc5b glycoproteins in horse saliva. Moreover, we also identified Muc19 from pig, cow and rat saliva; the saliva of cow and rat also contained Muc5b; however, due to the lack of pig Muc5b genomic sequence data, we were unable to identify Muc5b in pig saliva. Our results suggest that unlike human saliva, which contains MUC5B, cow, horse and rat saliva are a heterogeneous mixture of Muc5b and Muc19. The functional consequence of these species differences remains to be elucidated. PMID- 18426394 TI - Nitric oxide produced in Peyer's patches exhibits antiapoptotic activity contributing to an antimicrobial effect in murine salmonellosis. AB - Salmonella species normally infect hosts via the oral-fecal route. We previously reported that NO had potent host defense functions in murine salmonellosis, not only via a direct antibacterial effect but also because it was cytoprotective for infected host cells. Here, we used an oral route to infect iNOS-deficient mice infected with S. enterica serovar Typhimurium to further investigate the cytoprotective role of NO in preventing damage caused by Salmonella organisms in PP. Oral bacterial challenge (2 x 10(5) CFU, or >100 LD(50)) produced a more severe infection and greater lethality in iNOS-deficient mice than in iNOS competent mice. We used specific antibodies to S. enterica Typhimurium, neutrophils, iNOS, nitrotyrosine, and dendritic cells (CD11c-positive) in histochemical and immunohistochemical studies to examine infected PP tissues. S. enterica Typhimurium colonization in PP from iNOS-deficient mice was significantly higher than that in wild-type mice. Histochemical assays showed extensive cellular damage in PP. We then examined PP tissues for apoptosis by means of in situ TUNEL analysis and by measuring caspase-3 specific activity in tissue homogenates. Increased numbers of TUNEL-positive cells and severe granulomatous inflammation with increased infiltration of neutrophils and macrophages were observed during infection in iNOS-deficient mice compared with wild-type mice. iNOS-deficient mice had increased numbers of dendritic cells and significantly higher caspase-3-specific activity in PP. These data confirm that NO exerts its protective function not only through direct antibacterial action, but also by preventing apoptosis and thereby contributing to antimicrobial defense during salmonellosis. PMID- 18426395 TI - High level serum neutralizing antibody against HIV-1 in Chinese long-term non progressors. AB - NAb have been considered to be an important component of a protective immune response to HIV-1, yet the relationship between the capacity of HIV-1 NAb, the conserved neutralization epitopes and disease progression has been unclear. To gain a better understanding of the protective roles that NAb and conserved neutralization epitopes could play in LTNP, twenty-eight HIV-1-infected subjects were investigated by evaluation of the concentrations of HIV-1 NAb and conserved neutralization epitopes, using single-round PBMC neutralization assay and sequencing. Our study revealed that the concentration of NAb in LTNP was significantly higher than that in subjects with asymptomatic HIV (P < 0.05) and AIDS (P < 0.01). No amino acids substitutions were found in the conserved epitopes of the HIV-1 gp120 region in LTNP, whereas the viruses circulating both in persons with asymptomatic HIV and those with AIDS had amino acid substitutions in their conserved neutralization epitopes. This study suggests that high levels of NAb and stable epitopes in gp120 could play a crucial role in protection against disease progression. PMID- 18426396 TI - Effects of single-point amino acid substitutions on the structure and function neuraminidase proteins in influenza A virus. AB - In order to clarify the effect of amino acid substitutions on the structure and function of the neuraminidase (NA) protein of influenza A virus, we introduced single-point amino acid substitutions into the NA protein of the A/Tokyo/3/67 (H2N2) strain using PCR-based random mutation. The rate of tolerant random one amino acid substitutions in the NA protein was 47%. Rates of tolerant substitutions for the stalk and for the surface and inner portion of the head region of the NA protein were 79, 54, and 19%, respectively. Deleterious changes, such as those causing the NA protein to stop at the Golgi/endoplasmic reticulum, were scattered throughout the protein. On the other hand, the ratio of mutations with which the NA protein lost neuraminidase activity, but was transported to the cell surface, decreased in proportion to the distance from the structural center of enzyme active site. In order to investigate the effect of accumulated amino acid substitutions on the structural character of the N2NA protein during evolution, the same amino acid substitutions were introduced by site-directed mutagenesis at 23 homologous positions on N2 proteins of A/Tokyo/3/67, A/Bangkok/15/85 (H3N2), and A/Mie/1/2004 (H3N2). The results showed a shift, or discordance, in tolerance at some of the positions. An increase in discordance was correlated with the interval in years between virus strains, and the discordance rate was estimated to be 0.6-0.7% per year. PMID- 18426397 TI - PPE protein (Rv3425) from DNA segment RD11 of Mycobacterium tuberculosis: a novel immunodominant antigen of Mycobacterium tuberculosis induces humoral and cellular immune responses in mice. AB - Subtractive DNA hybridization of pathogenic M. bovis and BCG, and comparative genome-wide DNA microarray analysis of M. tuberculosis H37Rv and BCG identified several RD, designated as RD1 to RD16, between M. tuberculosis and M. bovis on the one hand and BCG on the other. These regions cover 108 ORF of M. tuberculosis H37Rv, and are deleted from all 13 BCG sub-strains currently used as anti tuberculosis vaccines in different parts of the world. In this study, we evaluated cellular and humoral immune response in C57BL/6 mice immunized with the PPE protein Rv3425, encoded by an ORF found in RD11 of M. tuberculosis. Rv3425 protein induced an increased Th1/Th2 type immune response in mice, characterized by an elevated concentration of IFN-gamma in antigen stimulated splenocyte culture and a strong IgG(1) antibody response. These results provide evidence on the immunogenicity of the PPE protein Rv3425 which, together with its reported immunodominant characteristics, imply that it may be a candidate for development of a vaccine for the control of TB. PMID- 18426398 TI - In silico prediction and immunological validation of common HLA-DRB1-restricted T cell epitopes of Candida albicans secretory aspartyl proteinase 2. AB - ABSTRACT Sap2 is the most abundant virulence factor expressed during Candida infection, and the principal protein known to induce antibody response during Candida infection in humans. Its role in T-cell activation however, has not yet been determined. Sequence analysis revealed that Sap2 contains two variable regions: Var1 and Var2. Computational predictions by the Hotspot Hunter program identified that Var1 contains three candidate T-cell epitopes, whereas Var2 contains four. Thirty-nine overlapping peptides of Sap2 were then synthesized, and tested for their ability to induce proliferation of PBMC from 12 donors. Peptides P11, P17 and P31 exhibited significantly higher proliferative indices when compared with those of other peptides or controls. P17 and P31 are located in the areas of prediction, while P11 is not. There were other peptides outside the prediction areas that could stimulate PBMC proliferation at low levels. Nevertheless, the proliferative noise caused by such peptides was ruled out by IL 2 ELISpot analysis. Only P17 and P31 were shown to induce clonal proliferation of IFN-gamma producing lymphocytes, suggesting that these two peptides contain T cell epitopes. P11, which stimulated IL-2 producing clones, contains a known B cell epitope. Interestingly, P17 and P31 elicited both Th1 and Th2 cell responses with significant numbers of IL-13 secreting clones in response to stimulation. Taken together, the computer-based T cell epitope prediction method could identify the immunogenic T cell epitopes of C. albicans Sap2 that promiscuously bind to the HLA-DRB1 supertype. PMID- 18426399 TI - A simple and rapid immunochromatographic test kit for rabies diagnosis. AB - In rabies endemic countries, funds and infrastructure are often insufficient to employ the approved gold standard for the definitive diagnosis of rabies: the direct fluorescent test. In the present study, two types (type 1 and 2) of an ICT kit were evaluated for detection of rabies. These were developed using monoclonal antibodies which recognize epitope II and III of the nucleoprotein of rabies virus. Both kits specifically detected all rabies virus strains and there was no cross reactivity with Lyssaviruses (Lagos, Mokola and Duvenhage), Rhabdovirus (VSV and Oita 296/1972) and other common canine-pathogenic viruses. In type 1, a single type of monoclonal antibody was used. It was capable of detecting recombinant nucleoprotein and showed sensitivity of 95.5% (42/44) and specificity of 88.9% (32/36) using brain samples from rabid dogs. In contrast, type 2 which was made of two different monoclonal antibodies had a lower sensitivity of 93.2% (41/44) and higher specificity of 100% (36/36). These ICT kits provide a simple and rapid method for rabies detection. They need neither cold chain for transportation nor complicated training for personnel. This diagnostic test is suitable for rabies screening, particularly in areas with a high prevalence of rabies and where the fluorescent antibody test is not available. PMID- 18426402 TI - Melanoma initiating cells - easy as ABC? PMID- 18426403 TI - A pigment evolution Kitlg. PMID- 18426404 TI - Human 'coat colour' genetics. PMID- 18426405 TI - Transforming growth factor-beta in cutaneous melanoma. AB - Transforming growth factor-beta (TGF-beta) plays a complex role during carcinogenesis. It may either act as a tumor suppressor through its broad antiproliferative potential or as a tumor promoter either via direct effects on tumor cell aggressiveness or indirectly by modulating stromal responses, angiogenesis and immune surveillance. Increased production of TGF-beta by cancer cells is often associated with tumor grade. Melanoma cells largely escape cell cycle arrest normally induced by TGF-beta in normal melanocytes, yet produce active TGF-beta and are capable of efficient transcriptional responses to the growth factor. In this review, we summarize the current knowledge about the role played by TGF-beta in melanoma progression and hypothesize about the appropriateness of targeting TGF-beta signaling for therapeutic intervention. PMID- 18426406 TI - An update on the honesty of melanin-based color signals in birds. AB - The control mechanisms and information content of melanin-based color signals in birds have generated much recent interest and controversy among evolutionary biologists. Initial experimental studies on this topic manipulated coarse metrics of an individual's condition (i.e. food intake, disease state) and failed to detect significant condition-dependence of melanin ornament expression. However, three new lines of research appear profitable and target specific factors associated with the production of melanin pigments. These include the role of (i) metals, (ii) amino acids, and (iii) testosterone and social interactions in shaping the extent and intensity of melanin-colored plumage patches. Here, I review recent studies of and evidence for these honesty-reinforcing mechanisms. PMID- 18426407 TI - Involvement of FKHRL1 in melanoma cell survival and death. AB - Melanoma is a highly aggressive tumour characterized by a strong resistance to apoptotic stimuli that give rise to a selective advantage for tumour progression and metastasis formation. Therefore, it is of paramount importance to better understand the mechanisms involved in this resistance to apoptosis. In this report, we focused our attention on FKHRL1, a member of the forkhead family of transcription factors, which controls expression of genes involved in cell cycle progression and apoptosis. In melanoma cells, we show that IGF1, which exerts pro survival properties, induces the phosphorylation and nuclear exclusion of FKHRL1 in a PI3K/AKT-dependent pathway. Moreover, we observe that over-expression of a non-phosphorylable mutant of FKHRL1 (FKHRL1-TM), constitutively localized to the nucleus, promotes apoptotic cell death of melanoma cells. Finally, we find that FKHRL1-TM decreases the expression of survivin, a member of the inhibitor of apoptosis protein and that survivin re-expression partially rescues the deleterious effects of FKHRL1. Taken together, these findings reveal, in melanoma cells, that endogenous FKHRL1 is a downstream target of the PI3K/AKT pathway and suggest that the phosphorylation of this transcription factor may be involved in the pro-survival effects of growth factors such as IGF1. On the other hand, forced nuclear localization of FKHRL1 decreases melanoma cell growth and may serve as a therapeutic strategy against melanoma. PMID- 18426408 TI - Identification of differentially expressed genes in spontaneously regressing melanoma using the MeLiM swine model. AB - Partial and some few cases of complete spontaneous regression have been observed in cutaneous melanoma patients but little is known about the molecular mechanisms involved. The Melanoblastoma-bearing Libechov Minipig (MeLiM) is a suitable animal model to study the phenomenon of spontaneous regression because MeLiM pigs exhibit naturally occurring melanomas which regress completely 6 months after birth. In this study, we used suppression subtractive hybridization (SSH) to identify molecular determinants of melanoma regression within swine melanoma tissues and melanoma cell cultures. Several markers involved in cell-adhesion, communication, -motility, signal transduction, negative regulation of cell proliferation, transport and immune response were identified that correlated with melanoma regression whereas the main genes involved in melanin synthesis showed a strong downregulation. For the most differentially expressed genes, we validated the results obtained by SSH with qRT-PCR and with immunohistochemistry for some of them (CD9, MITF, RARRES1). Most notable, for the first time in melanoma, we identified the retinoic acid responder 1 gene (RARRES1) as a main actor of the regression process in melanoma. This first gene expression study in swine melanoma regression, may contribute to the finding of new therapeutic targets for human melanoma treatment. PMID- 18426409 TI - Transcriptional profiling of melanocytes from patients with vitiligo vulgaris. AB - Vitiligo is a complex, polygenic disorder characterized by patchy loss of skin pigmentation due to abnormal melanocyte function. Both genetic and environmental etiological factors have been proposed for vitiligo and lack of molecular markers renders difficulties to predict development and progression of the disease. Identification of dysregulated genes has the potential to unravel biological pathways involved in vitiligo pathogenesis, facilitating discovery of potential biomarkers and novel therapeutic approaches. In this study, we characterized the transcriptional profile of melanocytes from vitiligo patients. Oligonucleotide microarrays containing approximately 16,000 unique genes were used to analyse mRNA expression in melanocytes from vitiligo patients and age-matched healthy controls. In total, 859 genes were identified as differentially expressed. A substantial number of these genes were involved in (i) melanocyte development, (ii) intracellular processing and trafficking of tyrosinase gene family proteins, (iii) packing and transportation of melanosomes, (iv) cell adhesion and (v) antigen processing and presentation. In conclusion, our results show a significantly different transcription profile in melanocytes from vitiligo patients compared with controls. Several genes of potential importance for the pathogenesis and development of vitiligo were identified. Our data indicate that autoimmunity involving melanocytes may be a secondary event in vitiligo patients caused by abnormal melanocyte function. PMID- 18426410 TI - LIGR, a protease-activated receptor-2-derived peptide, enhances skin pigmentation without inducing inflammatory processes. AB - The protease-activated receptor-2 (PAR-2) is a seven transmembrane G-protein coupled receptor that could be activated by serine protease cleavage or by synthetic peptide agonists. We showed earlier that activation of PAR-2 with Ser Leu-Ile-Gly-Arg-Leu-NH(2) (SLIGRL), a known PAR-2 activating peptide, induces keratinocyte phagocytosis and increases skin pigmentation, indicating that PAR-2 regulates pigmentation by controlling phagocytosis of melanosomes. Here, we show that Leu-Ile-Gly-Arg-NH(2) (LIGR) can also induce skin pigmentation. Both SLIGRL and LIGR increased melanin deposition in vitro and in vivo, and visibly darkened human skins grafted onto severe combined immuno-deficient (SCID) mice. Both SLIGRL and LIGR stimulated Rho-GTP activation resulting in keratinocyte phagocytosis. Interestingly, LIGR activates only a subset of the PAR-2 signaling pathways, and unlike SLIGRL, it does not induce inflammatory processes. LIGR did not affect many PAR-2 signaling pathways, including [Ca(2+)] mobilization, cAMP induction, the induction of cyclooxgenase-2 (COX-2) expression and the secretion of prostaglandin E2, interleukin-6 and -8. PAR-2 siRNA inhibited LIGR-induced phagocytosis, indicating that LIGR signals via PAR-2. Our data suggest that LIGR is a more specific regulator of PAR-2-induced pigmentation relative to SLIGRL. Therefore, enhancing skin pigmentation by topical applications of LIGR may result in a desired tanned-like skin color, without enhancing inflammatory processes, and without the need of UV exposure. PMID- 18426411 TI - Participation of nitric oxide in the color change induced by UV radiation in the crab Chasmagnathus granulatus. AB - The ability of UV radiation to stimulate color change in vertebrates is well known; however, the signaling pathway involved is not fully explained. Since nitric oxide (NO) is among the candidates for this role, in this study the participation of NO signaling in the pigment migration induced by UV radiation in melanophores of the crab Chasmagnathus granulatus was investigated. When the NO donor, SIN-1, was incubated with pieces of epidermis, there was an induction of a dose-dependent pigment dispersion (in vitro assays). When male adults were exposed to different doses of UVA and UVB, N(G)-nitro-l-arginine-methyl-ester, an NO synthase (NOS) blocker produced a decrease of the pigment dispersion induced by UV (in vivo assays). However, in similar assays, 2-phenyl-4,4,5,5 tetramethylimidazoline-1-oxyl-3-oxide, an NO scavenger, decreased only the pigment dispersion induced by UVA. Interestingly, buthionine sulfoximine did not produce any change in pigment dispersion induced by UVA (in vivo assays) and SIN 1 (in vitro assays). Our results using NADPH-diaphorase histochemistry and immunocytochemistry against nNOS indicated the production of NO by epidermal cells. In conclusion, we suggest that NO is a key molecule for the induction of pigment dispersion in the melanophores of Chasmagnthus granulatus, and also that NOS activation is a fundamental step for this process. PMID- 18426412 TI - The radioprotective properties of fungal melanin are a function of its chemical composition, stable radical presence and spatial arrangement. AB - Melanized microorganisms are often found in environments with very high background radiation levels such as in nuclear reactor cooling pools and the destroyed reactor in Chernobyl. These findings and the laboratory observations of the resistance of melanized fungi to ionizing radiation suggest a role for this pigment in radioprotection. We hypothesized that the radioprotective properties of melanin in microorganisms result from a combination of physical shielding and quenching of cytotoxic free radicals. We have investigated the radioprotective properties of melanin by subjecting the human pathogenic fungi Cryptococcus neoformans and Histoplasma capsulatum in their melanized and non-melanized forms to sublethal and lethal doses of radiation of up to 8 kGy. The contribution of chemical composition, free radical presence, spatial arrangement, and Compton scattering to the radioprotective properties of melanin was investigated by high performance liquid chromatography, electron spin resonance, transmission electron microscopy, and autoradiographic techniques. Melanin protected fungi against ionizing radiation and its radioprotective properties were a function of its chemical composition, free radical quenching, and spherical spatial arrangement. PMID- 18426413 TI - Correlation between melanogenic and catalase activity in in vitro human melanocytes: a synergic strategy against oxidative stress. AB - UV-induced DNA damage can lead to melanoma, the most dangerous form of skin cancer. Understanding the mechanisms employed by melanocytes to protect against UV is therefore a key issue. In melanocytes, catalase is the main enzyme responsible for degrading hydrogen peroxide and we have previously shown that that low basal levels of catalase activity are associated with the light phototype in in vitro and ex vivo models. Here we investigate the possible correlation between its activity and melanogenesis in primary cultures of human melanocytes. We show that while the total melanin concentration is directly correlated to the level of pigmentation, the more the degree of pigmentation increased, the lower the proportion of pheomelanin present. Moreover, in human melanocytes in vitro, catalase-specific mRNA, protein and enzymatic activity were all directly correlated with total cellular melanin content. We also observed that immediately after a peroxidative treatment, the increase in reactive oxygen species was inversely associated with pigmentation level. Darkly pigmented melanocytes therefore possess two protective strategies represented by melanins and catalase activity that are likely to act synergistically to counteract the deleterious effects of UV radiation. By contrast, lightly pigmented melanocytes possess lower levels of melanogenic and catalase activity and are therefore more susceptible to accumulate damage after UV exposition. PMID- 18426414 TI - The PTPN22-1858C>T (R620W) functional polymorphism is associated with generalized vitiligo in the Romanian population. AB - Generalized vitiligo is an autoimmune disorder of the skin in which autoimmune mediated destruction of melanocytes leads to depigmented patches of skin and overlying hair. The 1858C>T (R620W) functional polymorphism of the PTPN22 gene, which encodes lymphoid protein tyrosine phosphatase (Lyp), has been associated with susceptibility to a number of autoimmune disorders, including generalized vitiligo. The aim of this study was to test genetic association of the PTPN22 1858C>T variant and generalized vitiligo in a Romanian case-control cohort. We observed significant association of generalized vitiligo with the 1858T risk allele of PTPN22 [P = 0.0138; OR = 2.92 (1.21-7.03)], with significantly different distribution of PTPN22 1858C>T genotypes in cases versus controls [P = 0.036; OR = 2.69 (1.07-6.80)]. Our results provide evidence that the PTPN22 1858T allele contributes to risk of generalized vitiligo in European Caucasian populations, and underscores the importance of a genetically mediated autoimmune mechanism in the pathogenesis of vitiligo. PMID- 18426415 TI - Expression from a bacterial artificial chromosome containing the Dct gene locus. PMID- 18426417 TI - Recent progress in macular function self-assessment. AB - BACKGROUND: Patients that are deemed to be at risk for the development of choroidal neovascularization are frequently instructed to self-monitor their vision for symptoms that may signal the presence of choroidal neovascularization. Traditionally, the Amsler grid has been used for this purpose, but recent technological innovations have resulted in a number of potential alternatives that offer important advantages, including greater sensitivity, the ability to differentiate exudative from non-exudative disease, and quantitative analysis of test results. METHODS: The following technologies that could be employed for patient self-assessment of macular function in a home setting are reviewed: computerized Amsler grid, preferential hyperacuity perimetry, macular mapping test and noise-field campimetry. CONCLUSIONS: A number of technologies are currently available that could significantly improve the ability of patients at risk for the development of choroidal neovascularization to self-monitor their central visual field. PMID- 18426418 TI - A survey of the scope of therapeutic practice by UK optometrists and their attitudes to an extended prescribing role. AB - PURPOSE: Recent changes in medicines legislation in the UK have broadened the opportunities for optometrists to use and supply therapeutic drugs. We set out to investigate the current therapeutic practice of UK optometrists and to elicit their views on an extended prescribing role. METHODS: Members of the College of Optometrists were invited via email to take part in an online survey. The survey questions covered four areas: mode of practice, proximity and relationship to other providers of eye care, scope of current therapeutic practice and future plans regarding prescriber training. RESULTS: Of the 1,288 responses received (response rate 24%), over 90% were from optometrists working in community practice. Common, non-sight-threatening conditions were managed frequently or occasionally by between 69 and 96% of respondents. Blepharitis and dry eye were the most common (managed routinely by >70%). In terms of therapeutic agents used, large numbers of optometrists reported that they commonly supplied or recommended over-the-counter (non-prescription) drugs, particularly lubricants and anti allergic agents. However, fewer respondents supplied antibiotics (only 14% supplying chloramphenicol or fusidic acid frequently). Overall, relatively few respondents (14%) expressed no interest in undertaking further training for extended prescribing, although several barriers were identified, including cost and time taken for training, lack of remuneration and fear of litigation. CONCLUSION: Significant numbers of community optometrists are currently managing a range of common ocular conditions using a limited formulary. Enabling optometrists to train as independent prescribers will further develop this role, allowing greater use of their skills and providing patients with quicker access to medicines. PMID- 18426419 TI - Inattentional blindness and augmented-vision displays: effects of cartoon-like filtering and attended scene. AB - Augmented-vision devices that we are developing to aid people with low vision (impaired vision) employ vision multiplexing- the simultaneous presentation of two different views to one or both eyes. This approach enables compensation for vision deficits without depriving the wearers of their normal views of the scene. Ideally, wearers would make use of the simultaneous views to alert them to potential mobility hazards, without a need to divide attention consciously. Inattentional blindness, the frequent inability to notice otherwise-obvious events in one scene while paying attention to another, overlapping, scene, works against that sort of augmentation, so we are investigating ways to mitigate it. In this study, we filtered the augmented view, creating cartoon-like representations, to make it easier to detect significant features in that view and to minimise interference with the normal view. We reproduced a classic inattentional blindness experiment to evaluate the effect, and found that, surprisingly, edge filtering had no detectable effect - positive or negative - on the noticing of unexpected events in the unattended scene. We then modified the experiment to determine if the inattentional blindness was because of the confusion of overlaid views or simply a matter of attention, and found the latter to be the case. PMID- 18426420 TI - What do kids think about kids in eyeglasses? AB - PURPOSE: Previous studies have examined how people feel about others who wear glasses, but no studies of children have been published on the subject. We conducted the Children's Attitudes about Kids in Eyeglasses (CAKE) study to determine how children feel about other children who wear glasses. METHODS: Subjects compared a series of 24 picture pairs and answered six questions regarding which child ...he or she would rather play with, looks better at playing sports, appears smarter (more intelligent), appears nicer, looks more shy and looks more honest. The children in each pair of pictures differed by gender, ethnicity and spectacle wear. Logistic regression was performed to determine the probability and confidence interval that a subject would pick a particular child. RESULTS: Eighty subjects between the ages of 6 and 10 years participated. The average (+/-SD) age of the subjects was 8.3 +/- 1.3 years, 42 (53%) were females, 51 (64%) were whites, 21 (26%) were blacks, and 30 (38%) wore glasses. The spectacle wearer appeared smarter (0.66, CI = 0.60-0.71) and more honest (0.57, CI = 0.50-0.64), and children who wore glasses looked smarter regardless of whether the child picking wore glasses. Both boys (0.66, CI = 0.68-0.79) and girls (0.77, CI = 0.71-0.82) thought that boys looked better at playing sports. DISCUSSION: The old adage 'Boys never make passes at lasses who wear glasses' may be outmoded, but glasses may tend to make children look smarter and slightly more honest to their peers. PMID- 18426421 TI - The effect of positive-lens addition and base-in prism on accommodation accuracy and near horizontal phoria in Chinese myopic children. AB - The effect of positive-lens addition (0, +0.75, +1.50, +2.25, +3.00 D each eye) and base-in prism power (0, 1.5, 3 Delta each eye) on both near focusing errors and latent horizontal deviations was evaluated in 29 Chinese myopic children (age: 10.3 +/- 1.9 years, refractive error: -2.73 +/- 1.31 D). Accommodation response and phoria were measured by the Shin-Nippon auto-refractor (right eye) and Howell-Dwyer near phoria card at 33 cm with each of the 15 lens/prism combinations in random order. The initial accommodative error was -0.96 +/- 0.67 D (lag) and near phoria was -0.8 +/- 5.0 Delta (exophoria). The positive-lens addition decreased the accommodative lag but increased the exophoria as the power increased (e.g. up to -9.1 +/- 4.1 Delta with +3 D). A 6-Delta base-in prism totally controlled the exophoria induced by a +1.50 D addition (-0.3 +/- 4.3 Delta), but the accommodative lag was still considerable (-0.69 +/- 0.54 D). In the graphical analysis of the data, a lens addition of +2.25 D combined with a 6 Delta base-in prism minimized both the lag and lens-induced exophoria to -0.33 D and -2.4 Delta respectively (regression analysis). This lens and prism combination decreased the lens-induced exophoria by 4.5 Delta compared with that measured with +2.25 D alone (-2.4 Delta vs -6.9 Delta). The results suggest that incorporating near base-in prism when prescribing bifocal lenses for young progressing myopes with exophoria could reduce the positive lens-induced oculomotor imbalance. PMID- 18426422 TI - Developing an instrument to assess vision-related and subjective quality of life in children with intellectual disability: data collection and preliminary analysis in a Chinese population. AB - PURPOSE: Visual abnormalities are common in children and adolescents with intellectual disability, but it is not known whether these abnormalities impact on their quality of life. As a first step towards addressing this question, this study collects self-perceptions on visual performance and life quality in school children with intellectual disability. METHODS: Questionnaires were modified from those used to assess vision- and health-related quality of life in children with normal development, with guidance from a focus group of individuals who work or live with children with intellectual disability. The survey was conducted in a population of Chinese children with intellectual disability, using self completion and caregivers' responses. RESULTS: Responses from subjects showed comparable dimensions in vision-related quality of life to those revealed by previous studies in children with normal development. Some differences were apparent, however, and possible explanations for these are discussed. Our results showed good internal consistency and correlations between subscales within and across questionnaires. CONCLUSIONS: The assessment of vision-related quality of life is feasible in children and adolescents with at least mild-to-moderate intellectual disability. In this population of children with intellectual disability, perceived visual function is associated with quality of life. PMID- 18426423 TI - Usefulness of post-assessment reports in a paediatric low vision clinic: a questionnaire survey of parents and education professionals. AB - This survey aimed to evaluate how helpful parents and those working in the educational setting found written low vision assessment reports. A questionnaire survey was sent to 25 parents and to the corresponding and relevant member of the educational team for each of the 25 children. All were satisfied with the level of information contained in the report and agreed that the report was easy to understand and that the level of explanations given regarding visual function were clear. The main disparities between parents and teaching professionals were concerning the amount of medical terminology used in the report, with only 7% (n = 1) of those from the educational setting, but 30% (n = 6) of parents feeling there was too much. Adaptations were also more likely to have been implemented in the educational setting [82% (n = 11)] as compared with 62% (n = 16) in the home. Eighty-three per cent (n = 43) of respondents reported that low vision aids were used at either home or school on at least a weekly basis. PMID- 18426424 TI - Performance of colour-deficient people on the Holmes-Wright lantern (type A): consistency of occupational colour vision standards in aviation. AB - INTRODUCTION: The Holmes-Wright lantern type A (H-W A) is an occupational colour vision test used by the UK Civil Aviation Authority (CAA) and approved by Joint Aviation Requirements (JAR) to select aircrew. Pass, to obtain a CAA Class 1 Aviation medical certificate, can be achieved at three stages of the examination. The Commission Internationale d'Eclairage (CIE) recommends that the Falant pass criteria are used with all approved lanterns. A pass to obtain CIE Colour Vision Standard 2 can be achieved at two stages of the examination. This study examines the consistency of these pass criteria. METHODS: One hundred and twenty-five men with red-green colour deficiency were examined. All subjects completed three runs of the nine colour pairs shown on the H-W A at high brightness, in photopic and scotopic viewing. RESULTS: Ten of 78 deuteranomalous trichromats examined passed to obtain a CAA Class 1 Aviation medical certificate at the first stage of the examination but only two of these subjects were successful at all three stages. Seventeen deuteranomalous trichromats passed to obtain CIE Colour Vision Standard 2 in photopic viewing and 20 subjects (one protanope and 19 deuteranomalous trichromats) passed in scotopic viewing. Only 50% of subjects who passed at the first stage of the examination were also successful at the second stage in either viewing condition. Ten deuteranomalous trichromats passed to obtain CIE Colour Vision Standard 2 in both photopic and scotopic viewing. Forty-three per cent of subjects made red-green errors and 79% made red-white errors at some stage of the examination. CONCLUSIONS: The staged pass criteria used by the CAA and the CIE lack internal consistency when applied to the H-W A. Colour-deficient people who pass to a standard at the first stage of the examination are unlikely to be successful if the examination is continued. The staged pass criteria do not identify individuals with superior colour discrimination ability and it is difficult to justify selection of personnel for high risk occupations in aviation on this basis. PMID- 18426425 TI - Pass rates for the Farnsworth D15 colour vision test. AB - INTRODUCTION: The Farnsworth D15 test (D15) is used worldwide to select applicants for employment in occupations which require good colour vision. People with slight colour deficiency are intended to pass the D15 and people with significant (moderate/severe) colour deficiency to fail. METHODS: Pass rates were determined for 710 adult males with red-green colour deficiency using three different pass criteria in general use. RESULTS: Forty-six per cent of subjects were successful when the pass criterion was a circular results diagram (one single transformation of adjacent hues was accepted as a pass), 53% passed when one red-green isochromatic error was allowed and 60% passed when two red-green isochromatic errors were permitted. The pass rate for 200 dichromats was 1.5% on a circular diagram, 3% on one red-green error and 6% on two red-green errors. Protans made fewer errors than deutans and more protans than deutans were successful when either one or two red-green crossings were permitted as a pass. CONCLUSION: A circular results diagram is the preferred pass criterion. This criterion most nearly fulfils the aim of the test to fail all dichromats and people with significant protanomalous and deuteranomalous trichromatism. A circular diagram is also easy to interpret consistently. Re-examination is recommended if there are only one or two red-green isochromatic error lines across the results diagram. This gives individuals with borderline slight/moderate colour deficiency an opportunity to pass at the second attempt. PMID- 18426426 TI - The validity of the Jessen formula in overnight orthokeratology: a retrospective study. AB - PURPOSE: To investigate the validity of the Jessen formula, with a compression factor of 0.75, in determining the back optic zone radius (BOZR) of an orthokeratology (ortho-k) lens for myopic reduction [i.e. BOZR = flattest K- (target reduction + 0.75)]. METHODS: One hundred and twenty-three consecutive ortho-k patient files from the Optometry Clinic of The Hong Kong Polytechnic University were reviewed. Pertinent data at the preliminary visit and at the morning visit after 2 weeks of lens wear for 63 patients who fulfilled the inclusion criteria were retrieved for analysis. All patients were either fitted with DreimLens (DreimLens Taiwan, Taiwan Macro Vision Group, Taiwan, China) or eLens (E&E Optics Ltd., Hong Kong SAR, China) designs. Only data from the right eye were analysed. The validity of the Jessen formula was evaluated by comparing the equation of the plot of myopic reduction attempted (based on the Jessen formula) and myopic reduction achieved (based on subjective refraction). RESULTS: The Jessen formula was found to underestimate the intended target of myopic reduction following ortho-k. The results suggested that the formula should be revised to BOZR = flattest K- (1.23 target reduction + 1.27). CONCLUSION: If the intention is to overcorrect the ortho-k patient by 0.75 D to allow for regression during the daytime, the Jessen formula with a compression factor of 0.75 is not valid to determine the BOZR of the ortho-k lens designs. PMID- 18426427 TI - Optimal spherical focus in the peripheral retina. AB - It is well known that, in most eyes, astigmatism increases with the field angle. A simple reduced-eye model is used to demonstrate that, for point imagery in the peripheral retina, the combination of oblique astigmatism with elliptical entrance and exit pupils means that the retinal image with the optimal rotational symmetry is not necessarily at a focus corresponding to a 'best-sphere' correction. Equations are derived for the position of focus at which a circularly symmetric blur patch is obtained and for the dimensions of the patch in this and other image planes. Ray tracing through a wide-angle schematic eye is used to explore the validity of the simple model. It is shown that although the latter gives good predictions of retinal imagery for very small entrance pupils, it becomes less valid for larger, more realistic pupil diameters, due to the increasing importance of the effects of higher-order aberrations. Nevertheless, the simple model can still yield useful insights into through-focus, peripheral optical imagery. PMID- 18426428 TI - On the compensation of horizontal coma aberrations in young human eyes. AB - The nature of the compensation of horizontal coma (Z(3)(1)) between optical elements of the human eye has been studied and the compensative mechanism has been attributed to a passive process linked to angle kappa of the eye. We measured the horizontal coma in the anterior cornea, the whole eye and the internal optics for 221 young subjects. Thirty-three eyes with minimum angle lambda and 53 eyes with relatively large angle lambda were selected from these eyes to test the hypothesis that horizontal coma compensation is linked to angle kappa. Significant horizontal coma in the anterior cornea was observed for the group with minimum angle lambda in both the right (-0.12 +/- 0.07 microm) and left eyes (0.12 +/- 0.10 microm), and this was well compensated by the internal optics so that the level of horizontal coma in the whole eye over a 6-mm pupil size was very low (-0.05 +/- 0.07 microm for OD and 0.02 +/- 0.08 microm for OS). The horizontal coma in the anterior cornea was significantly correlated to the horizontal coma in the internal optics for both the right and the left eye. The results suggest that there is another source of horizontal coma, in addition to that linked to angle kappa, in the anterior cornea, and also a new compensative mechanism to balance the corneal coma, perhaps in the posterior cornea or the lens. PMID- 18426429 TI - Monocular fixation with the optic nerve head: a case report. AB - PURPOSE: To document and discuss the case of a patient with left esotropia (ET) who uses the left optic nerve head (ONH) for monocular 'fixation'. CASE REPORT: The patient was an 80-year-old male with left ET from early childhood. Retinal tracking monocular fixation measurements with a Nidek MP-1 revealed stable fixation within the left ONH area. In an attempt to challenge the initial observation, further assessments of fixation were performed with a smaller target size and requiring various gaze positions. MP-1 fixation data showed remarkably stable monocular fixation (+/-1 degrees over 30 s) mostly within the left ONH for all the target sizes and positions of gaze tested. Additional clinical binocular evaluations showed concomitant left ET approximately 28 Delta, no movement with cover test regardless of fixation target and no significant monocular motility restrictions. Visuoscopy also revealed fixation at the left ONH. There was a strong family history of ET, but none of the other affected descendants tested (n = 3) demonstrated the same behaviour. CONCLUSIONS: This is the first report documenting an abnormally developed monocular ocular motor system, with principal visual direction and zero retinomotor value shifted from the fovea to the ONH. We do not believe that there is any direct visual input from the ONH. The patient may use visual information obtained by glancing with peri-papillary areas to determine the target position (although this was largely ruled out), or obtain position information from the average luminance produced by scattered light around the ONH margin. The abnormal oculocentric direction might then be combined with extraretinal information (efferent copy or extraocular muscle proprioception) of the eye location in the orbit to stabilize the fixation. This patient does not have the blind spot syndrome (Swan, 1948). We propose the use of a retinal perimeter for documentation of eccentric fixation in strabismus. PMID- 18426430 TI - Meares-Irlen syndrome - a need for increasing awareness in the general public. PMID- 18426434 TI - Electrocardiographic alterations during hyperinsulinemic hypoglycemia in healthy subjects. AB - BACKGROUND: We evaluated the arrhythmogenic potential of hypoglycemia by studying electrocardiographic (ECG) changes in response to hyperinsulinemic hypoglycemia and associated sympathoadrenal counterregulatory responses in healthy subjects. METHODS: The study population consisted of 18 subjects, aged 30-40 years. Five minute ECG recordings and blood samplings were performed at baseline and during the euglycemic and hypoglycemic hyperinsulinemic clamp studies. PR, QT, and QTc intervals of electrocardiogram and ECG morphology were assessed from signal averaged ECG. RESULTS: Although cardiac beat interval remained unchanged, PR interval decreased (P < 0.01) and QTc interval (P < 0.001) increased in response to hyperinsulinemic hypoglycemia. Concomitant morphological alterations consisted of slight increases in R-wave amplitude and area (P < 0.01 for both), significant decreases in T-wave amplitude and area (P < 0.001 for both), and moderate ST depression (P < 0.001). Counterregulatory norepinephrine response correlated with amplification of the R wave (r =-0.620, P < 0.05) and epinephrine response correlated with flattening of the T wave (r =-0.508, P < 0.05). CONCLUSIONS: Hyperinsulinemic hypoglycemia with consequent sympathetic humoral activation is associated with several ECG alterations in atrioventricular conduction, ventricular depolarization, and ventricular repolarization. Such alterations in cardiac electrical function may be of importance in provoking severe arrhythmias and "dead-in-bed" syndrome in diabetic patients with unrecognized hypoglycemic episodes. PMID- 18426435 TI - Clinical implications of QRS duration and QT peak prolongation in patients with suspected coronary disease referred for elective cardiac catheterization. AB - BACKGROUND: The electrocardiogram (ECG) remains a simple, universally available, and prognostically powerful investigation in heart failure, and acute coronary syndromes. We sought to assess the prognostic utility of clinical, angiographic, and simple ECG parameters in a large cohort of patients undergoing elective cardiac catheterization (CC) for known or suspected coronary artery disease. METHODS: Consecutive consenting patients undergoing CC for coronary disease were enrolled at a single tertiary center. Patient data, drug therapy, catheter reports, and ECG recordings were prospectively recorded in a validated electronic archive. The primary outcome measure was death or nonfatal myocardial infarction (MI) over 1 year or until percutaneous or cardiac surgical intervention. Independent prognostic markers were identified using the Cox proportional hazard model. RESULTS: A total of 682 individuals were recruited of whom 17(2.5%) died or suffered a nonfatal MI in 1 year. In multivariate analysis QRS duration (ms) (HR 1.03 95% CI 1.01-1.05, P = 0.003), extent of coronary disease (HR 2.01 95% CI 1.24-3.58, P = 0.006), and prolonged corrected QT peak interval in lead I (HR 1.02 95% CI 1.00-1.03, P = 0.044) were independently associated with death or nonfatal MI. Receiver-operator characteristic (ROC) analysis for the multivariate model against the primary end point yielded an area under the curve of 0.759 (95% CI 0.660-0.858), P < 0.001. CONCLUSIONS: QRS duration and QT peak are independently associated with increased risk of death or nonfatal MI in stable patients attending for coronary angiography. PMID- 18426436 TI - The prognostic value of T wave amplitude in lead aVR in males. AB - BACKGROUND: Since there is an uncertainty regarding which of the 12 leads provides the most information, we investigated the association between repolarization phenomenon in all of the 12 leads and cardiovascular (CV) mortality. METHODS: Retrospective cohort study was performed at Palo Alto Veterans Affairs Medical Center, Palo Alto, California, which included 24,270 consecutive male veterans with ECGs obtained for clinical reasons from 1987 to 2000. Analysis of computerized 12-lead resting ECGs was performed of all subjects excluding inpatients, patients with atrial fibrillation, WPW, QRS duration > 120 ms, and paced rhythms. Average follow-up was 7.5 years during which time there were 1859 CV deaths. RESULTS: While ST segment measurements in aVR were univariately predictive of CV death, T wave amplitude superseded them in multivariate survival analysis. In addition, T wave amplitude in aVR outperformed repolarization measurements in all other leads as well as other ECG criteria (Q waves, damage scores, LVH) for predicting CV mortality. As T wave amplitude became less negative in aVR, there was a progressive increase in relative risk (RR). When the T waves in aVR had a positive deflection (i.e., upward pointing) the RR for CV death was 5.0. CONCLUSIONS: T wave amplitude in lead aVR is a powerful prognostic marker for estimating risk of CV death. Upward pointing T waves (a simple visual criterion) was prevalent (7.3% of a clinical population) and was associated with an annual CV mortality of 3.4% and a risk of five times. PMID- 18426437 TI - Autonomic markers as predictors of nonfatal acute coronary events after myocardial infarction. AB - BACKGROUND: Autonomic markers, such as heart rate variability (HRV), heart rate turbulence (HRT), and baroreflex sensitivity (BRS) provide information on the risk of all-cause mortality after an acute myocardial infarction (AMI), but their value in predicting nonfatal cardiac events is not well known. METHODS: A consecutive series of 675 patients with an AMI were followed up to 30 months. At baseline, the patients underwent a 24-hour Holter recording, and assessment of BRS using phenylephrine test. Several parameters of HRV and HRT were determined. RESULTS: After the follow-up, 98 patients (15%) had a nonfatal acute coronary event. Among the studied variables, the short-term scaling exponent alpha1 (P = 0.002), power-law slope beta (P = 0.008), low-frequency component of HRV power spectrum (P < 0.001), turbulence slope (P < 0.001), and BRS (P < 0.001) had the strongest association with the occurrence of nonfatal acute coronary events in univariate comparisons. After adjustment with relevant clinical variables (such as age, gender, ejection fraction, functional class, medication, diabetes) in the Cox proportional hazards model, alpha1 and beta remained as statistically significant predictors of nonfatal acute coronary events (HR = 2.0 [1.2-3.2, 95% CIs, P = 0.006] for alpha1 < or = 1.025), (HR = 1.9 [1.2-3.1, P = 0.008] for beta < or =-1.507). CONCLUSION: Several autonomic markers provide information on the risk of recurrent nonfatal coronary events after an AMI. Altered fractal heart rate behavior seems to be the strongest independent predictor of such events. PMID- 18426438 TI - Adverse effects of continuous ventricular pacing in patients with slower atrial fibrillation and normal left ventricular systolic function. AB - BACKGROUND: Both heart rate irregularity during chronic atrial fibrillation (AF) and ventricular desynchronization imposed by ventricular pacing may compromise ventricular function. We investigated whether heart rhythm regularization achieved through ventricular overdrive pacing (VP) gives additional benefit over rate control alone in patients with AF. METHODS: We studied 27 patients (mean age 72 +/- 7 years) with AF and normal left ventricular (LV) systolic function who were implanted with a common VVIR pacemaker. Cardiac function was assessed by using serial echocardiographic conventional, tissue Doppler imaging (TDI) and color M-Mode (CMM) examinations, together with B-type natriuretic peptide (BNP) measurements. Baseline data were obtained during AF (mean heart rate 58 +/- 5 beats/minute) with the pacemakers programmed to ventricular mere back-up pacing. These data were compared to the corresponding measurements following a 2-week VP period after the devises had been programmed to a lower rate of 70 beats/min, ensuring most of the time continuing VP. RESULTS: Continuous VP compared to AF, reduced the LV cardiac index (2.28 +/- 0.44 l/min/m(2) vs 2.33 +/- 0.39 l/min/m(2), P < 0.05), increased the LV end-systolic volume (38 +/- 14 mL vs 35 +/- 11 mL, P < 0.05), and decreased the TDI-derived systolic and diastolic mitral velocity (8.1 +/- 1.8 cm/s vs 8.3 +/- 1.6 cm/s, and 8.1 +/- 1.8 cm/s vs 8.3 +/- 1.6 cm/s, respectively, both P < 0.05) and the CMM-derived transmitral early diastolic flow propagation velocity (37.6 +/- 9.2 vs 41.5 +/- 9.7, P < 0.05). Following VP, both ratios E/Ea and E/Vp showed a trend toward increase (P = NS), whereas BNP rose up to 25.5% (median value, from 111 pg/mL to 165 pg/mL, P < 0.01). CONCLUSION: VP may be considered disadvantageous compared to slower AF. PMID- 18426439 TI - Effects of cigarette smoking on heart rate variability and plasma N-terminal pro B-type natriuretic peptide in healthy subjects: is there the relationship between both markers? AB - BACKGROUND: Cigarette smoking increased the risk of acute cardiac events related with endothelial dysfunction and increased sympathetic activity. Impaired autonomic nervous activity is recognized as a considerable symptom of cardiac dysfunction and is strongly associated with increased risk overall mortality. METHODS: A total of 75 healthy habitual smokers (40 female, 35 male, mean age 36.5 +/- 8.5 years), and 73 non-smokers subjects (45 female, 28 male, mean age 34.6 +/- 7.2 years) were studied. LF and LF/HF ratio were significantly higher in smokers than in non-smokers. On the contrary, SDNN, SDANN, RMSSD, and HF values were lower in smokers compared to those in non-smokers. Not the duration of smoking but the number of cigarettes smoked per day was correlated with the HRV parameters and NT-pro BNP. Furthermore, the average levels of NT-pro BNP were found to be positively correlated with LF, LF/HF and inversely correlated with SDNN, SDANN, RMSSD and HF. RESULTS: As a result, smoking impaires sympathovagal balance and decreases the heart rate variability in healthy subjects. And even a one cigarette smoking leads to overt sympathetic excitation. Furthermore, smoking results in an increase in NT-proBNP levels and the changes in adrenergic nervous system and NT-proBNP levels are well correlated. CONCLUSION: These findings could contribute to the higher rate of cardiovascular events in smokers. PMID- 18426440 TI - Development and validation of diagnostic criteria for atrial flutter on the surface electrocardiogram. AB - BACKGROUND: There are no universally accepted ECG diagnostic criteria for atrial flutter (AFL), making its differentiation from "coarse" atrial fibrillation (AF) difficult. METHODS: To develop diagnostic criteria for AFL, we examined two sets of ECGs. Set 1 consisted of 100 ECGs (50 AF, AFL) with diagnoses confirmed by intracardiac recordings. Criteria evaluated were presence of F waves in the frontal plane leads, F waves in V(1), sawtooth F waves, rate, and regularity of ventricular response. Set 2 included 200 ECGs taken from the hospital database each of which had already been interpreted by a cardiologist as either AF (n = 100) or AFL (n = 100). Set 2 was blindly read by electrophysiologists whose consensus-diagnoses were compared to the diagnoses made by using the best criteria identified from the Set 1 data. RESULTS: The criteria of frontal plane F waves, regular or partially regular ventricular response, and their combination had sensitivities of 92%, 98%, and 90% and specificities of 100%, 78%, and 100% in Set 1 for the diagnosis of AFL. In Set 2, concordance of electrophysiologist and cardiologist diagnoses was only 84%. The criteria of frontal plane Fwaves, regular or partially regular ventricular response, and their combination resulted in concordances with the cardiologist diagnoses of 85%, 85%, and 82% and with the electrophysiologist-consensus diagnoses of 90%, 89%, and 94% (P < 0.001). CONCLUSIONS: The criteria of frontal plane F waves and regular or partially regular ventricular response aid in the proper diagnosis of AFL. Because management strategies may differ for AF and AFL, it is important to adopt a more rigorous diagnostic approach. PMID- 18426441 TI - Comparison of wavelet transform modulus maxima and multifractal detrended fluctuation analysis of heart rate in patients with systolic dysfunction of left ventricle. AB - BACKGROUND: In recent years the WTMM (wavelet transform modulus maxima) and MDFA (multifractal detrended fluctuation analysis) methods have become widely used techniques for the determination of nonlinear, multifractal heart rate (HR) dynamics. The purpose of our study was to compare multifractal parameters of heart rate calculated using both methods in a group of 90 patients with reduced left ventricular systolic function (rlvs group) and in a group of 39 healthy persons (nsr group). METHODS: For each subject from the rlvs group (LVEF < or =40%) and the nsr group, a 24-hour ECG Holter monitoring was performed. The width of the multifractal spectrum and global Hurst exponent were calculated by means of WTMM and MDFA methods for 5-hour daytime and nighttime subsets. RESULTS: The width of the multifractal spectrum was significantly lower and the Hurst exponent was significantly higher in rlvs group in comparison to nsr group both during diurnal activity and nocturnal rest according to MDFA and only during diurnal activity according to WTMM method. In both groups we observed significant differences of the multifractal spectrum width and the global Hurst exponent between the nighttime and daytime recordings. CONCLUSIONS: MDFA seems to be more sensitive as compared with WTMM method in differentiation between multifractal properties of the heart rate in healthy subjects and patients with left ventricular systolic dysfunction. PMID- 18426442 TI - Is wider worse? QRS duration predicts cardiac mortality in patients with right bundle branch block. AB - BACKGROUND: Increasing QRS duration may be of prognostic significance in patients with right bundle branch block (RBBB) and may assist in predicting overall cardiovascular risk. METHODS: To test this hypothesis, we examined the Computerized Patient Records of patients with complete and persistent RBBB. Primary and secondary end points were all-cause and cardiac mortality. The effects of QRS duration on death rates were analyzed using the Cox proportional hazards regression model (P < 0.05). We identified 52,852 patients with EKGs and selected all those with diagnosis of RBBB (QRS > or = 120 ms) between January 2000 and January 2004. Some patients had EKG records confirming RBBB since 1987. The QRS durations were categorized into four groups: 120-129, 130-139, 140-149, and > or = 150 ms. RESULTS: A total of 997 (1.9%) patients (mean age 68.9 +/- 10 years) with RBBB were followed for 1-226 (median 45) months. All-cause mortality occurred in 344 (34.5%), cardiac deaths in 59 (5.9%), noncardiac in 191 (19.2%), and unknown causes in 94 (9.4%) patients. Mean left ventricular EF for cardiac patients was 38 +/- 15%. In patients with cardiac deaths, QRS duration was associated with increased morality (P < 0.007). For every 10 ms increase in QRS duration, the risk of death rose by 26.6%. The effect of QRS duration on all cause mortality was not statistically significant (P < 0.43). CONCLUSION: Increasing QRS duration was an independent predictor of cardiac mortality in patients with RBBB, but had no influence on all-cause mortality. QRS duration has added prognostic information to the presence of right bundle branch block. PMID- 18426443 TI - Resting magnetocardiography predicts 3-year mortality in patients presenting with acute chest pain without ST segment elevation. AB - OBJECTIVE: Magnetocardiography (MCG) as a noninvasive, noncontact and risk-free diagnostic method predicts ischemic coronary artery disease (CAD) in patients with acute chest pain at admission with high accuracy. However, it remains unclear whether MCG findings can add prognostic information. METHOD: A cohort of 402 consecutive patients presenting at the intensive care unit (ICU) with acute chest pain without ST segment elevation (NSTEMI) were included in a prospective registry. In order to prove the prognostic value of MCG a head-to-head comparison of the admission MCG, ECG, TnI, and ECHO tests was made. RESULTS: In 43 patients (10.7%) the MCG could not be analyzed due to insufficient signal-to-noise ratio. Complete follow-up over a period of up to 3 years was obtained in 355 out of the 359 patients (98.9%). Age at admission was 67.2 +/- 10.3 years, 59.7% males. In the group of patients with an abnormal MCG at admission, 43 out of 249 patients (17.3%) died in the follow-up period, while in the group of patients with a normal MCG at admission only 4 out of 106 patients died (3.77%). The relative risk was 4.58 (95% confidence intervals: 1.68-12.42). A multivariate regression analysis revealed the highest mortality risk for patients with diabetes mellitus and an abnormal MCG at admission (RR = 18.0; 95% CI: 2.49-133.3). CONCLUSION: Resting MCG at hospital admission predicts 3-year mortality in patients presenting with acute chest pain without ST segment elevation in the ECG. MCG seems to be valuable in identifying chest pain patients at highest risk. PMID- 18426444 TI - Single nucleotide polymorphisms and haplotype of four genes encoding cardiac ion channels in Chinese and their association with arrhythmia. AB - BACKGROUND: Many studies revealed that variations in cardiac ion channels would cause cardiac arrhythmias or act as genetic risk factors. We hypothesized that specific single nucleotide polymorphisms in cardiac ion channels were associated with cardiac rhythm disturbance in the Chinese population. METHOD: We analyzed 160 nonfamilial cardiac arrhythmia patients and 176 healthy individuals from which 81 individuals were selected for association study, and a total of 19 previously reported SNPs in four cardiac ion channel genes (KCNQ1, KCNH2, SCN5A, KCNE1) were genotyped. RESULTS: The frequency of KCNQ1 1638G>A, as well as the haplotype harboring KCNQ1 1638A, KCNQ1 1685 + 23G and 1732 + 43T (haplotype AGT) was significantly higher in healthy controls than in arrhythmia patients. This finding implicated that this haplotype (AGT) might be a protective factor against arrhythmias. CONCLUSIONS: Our study provided important information to elucidate the effect of SNPs of cardiac ion channel genes on channel function and susceptibility to cardiac arrhythmias in Chinese population. PMID- 18426445 TI - Baroreflex sensitivity: measurement and clinical implications. AB - Alterations of the baroreceptor-heart rate reflex (baroreflex sensitivity, BRS) contribute to the reciprocal reduction of parasympathetic activity and increase of sympathetic activity that accompany the development and progression of cardiovascular diseases. Therefore, the measurement of the baroreflex is a source of valuable information in the clinical management of cardiac disease patients, particularly in risk stratification. This article briefly recalls the pathophysiological background of baroreflex control, and reviews the most relevant methods that have been developed so far for the measurement of BRS. They include three "classic" methods: (i) the use of vasoactive drugs, particularly the alpha-adrenoreceptor agonist phenylephrine, (ii) the Valsalva maneuver, which produces a natural challenge for the baroreceptors by voluntarily increasing intrathoracic and abdominal pressure through straining, and (iii) the neck chamber technique, which allows a selective activation/deactivation of carotid baroreceptors by application of a negative/positive pressure to the neck region. Two more recent methods based on the analysis of spontaneous oscillations of systolic arterial pressure and RR interval are also reviewed: (i) the sequence method, which analyzes the relationship between increasing/decreasing ramps of blood pressure and related increasing/decreasing changes in RR interval through linear regression, and (ii) spectral methods, which assess the relationship (in terms of gain) between specific oscillatory components of the two signals. The limitations of the coherence criterion for the computation of spectral BRS are discussed, and recent proposals for overcoming them are presented. Most relevant clinical applications of BRS measurement are finally reviewed with particular reference to patients with myocardial infarction and heart failure. PMID- 18426446 TI - It is not always digitalis: bidirectional ventricular tachycardia in left ventricular hypertrophy. PMID- 18426447 TI - Ultrafiltration for heart failure: assessment of intravascular volume status through ECG modifications. PMID- 18426450 TI - Research on couples. PMID- 18426451 TI - Short- and long-term impact of critical illness on relatives: literature review. AB - AIM: This paper is a report of a literature review undertaken to identify the short- and long-term impact of critical illness on relatives. BACKGROUND: Patients in intensive care can experience physical and psychological consequences, and their relatives may also experience such effects. Although it is recognized that relatives have specific needs, it is not clear whether these needs are always met and whether further support is required, particularly after intensive care. DATA SOURCES: The following databases were searched for the period 1950-2007: Medline, British Nursing Index and Archive, EMBASE, CINAHL, PsycINFO and EMB Reviews--Cochrane Central Register of Clinical Trials. SEARCH METHODS: Search terms focused on adult relatives of critically ill adult patients during and after intensive care. Recurrent topics were categorized to structure the review, i.e. 'relatives needs', 'meeting relatives' needs', 'interventions', 'satisfaction', 'psychological outcomes' and 'coping'. RESULTS: Studies have mainly identified relatives' immediate needs using the Critical Care Family Needs Inventory. There are few studies of interventions to meet relatives' needs and the short- and long-term effects of critical illness on relatives. CONCLUSION: Despite widespread use of the Critical Care Family Needs Inventory, factors such as local or cultural differences may influence relatives' needs. Relatives may also have unidentified needs, and these needs should be explored. Limited research has been carried out into interventions to meet relatives' needs and the effects of critical illness on their well-being, yet some relatives may experience negative psychological consequences far beyond the acute phase of the illness. PMID- 18426452 TI - 'As required' medication regimens for seriously mentally ill people in hospital. PMID- 18426453 TI - Bed rest for acute uncomplicated myocardial infarction. PMID- 18426454 TI - Nurses' leaving intentions: antecedents and mediating factors. AB - AIM: This paper is a report of a study to investigate how nurses' work values, perceptions of environmental characteristics, and organizational commitment are related to their leaving intentions. BACKGROUND: Nurse leaving is a serious international problem as it contributes to the nursing shortage that threatens the welfare of society. The characteristics of nurses, the work environment and nurses' feelings towards their jobs (or organizations) have an impact on their leaving intentions. METHOD: A convenience sample of 849 Registered Nurses was recruited from three public hospitals in the central-west region of Japan during 2006 and 319 completed questionnaires were returned (response rate 39%). Data were analysed using regression analysis. FINDINGS: Nurses' work values and their perceptions of their workplace environment interacted to influence leaving intentions. When there was a match between the importance nurses placed on being able to challenge current clinical practices and the number of the actual opportunities to do so, leaving intentions were low. When there was a mismatch, intention to quit the job became stronger. In addition, organizational commitment intervened between nurses' perceptions of the match in clinical challenges and leaving intention. CONCLUSION: Nurses' leaving intentions, deserve extensive exploration of their causes. Such exploration should include attending to both nurses' needs and organizational characteristics, investigating how the match between them could affect nurses' leaving intention, and exploring factors that intervene between nurses' perceptions of the match and leaving intention. PMID- 18426455 TI - 'Veiling sexualities': a grounded theory of mental health nurses responses to issues of sexuality. AB - AIM: This paper is a report of a study to develop a grounded theory explaining how mental health nurses respond to issues of sexuality in clinical practice. BACKGROUND: The history of sexuality and people with mental health problems has largely been a history shrouded in misunderstanding, stigma, myth and negativity. However, individuals with mental health problems may experience sexuality and relationship difficulties related to their life experiences, mental illness, or its treatment. METHODS: Grounded theory was the methodology used for the study. Interviews were conducted in 2005-2006 with 27 mental health nurses working an urban area in the Republic of Ireland. Data were analysed using the concurrent processes of constant comparative analysis, data collection, theoretical sampling and memo writing. FINDINGS: The core category to emerge from the data was 'veiling sexualities'. This refers to participants' accounts of how they responded to the sexuality dimension of clients' lives. Participants' main concerns about sexuality were related to feelings of personal and professional vulnerability, due to a lack of competence, comfort and confidence in this area. The theory highlights the manner in which nurses perpetuate practices that marginalize, discriminate and socially exclude clients as 'sexual citizens'. CONCLUSION: The theory of Veiled Sexualities may facilitate acknowledgement of the presence of sexuality in all nurse-client encounters, and promote a discourse on the sexual rights of people experiencing mental distress among mental health nurses and all involved in the delivery of mental health services. PMID- 18426456 TI - Types of nursing knowledge used to guide care of hospitalized patients. AB - AIM: This paper is a report of a study to identify the types of nursing knowledge used to guide care of hospitalized patients. BACKGROUND: The history of nursing knowledge is discerned in three distinct moments. The first historical moment conceives nursing knowledge as the acquisition of a set of descriptive rules, the second as the development of dualist explanatory theories and the third as the production of critical and/or integrative understandings. It remains unclear how these different types of knowledge are implemented in practice and how they affect the care of hospitalized patients. METHOD: A secondary qualitative analysis was conducted in 2007 on original data collected in 2002. The data were read with focus on the knowledge used by participants to confront practice situations. They were interpreted, classified and indexed to identify types of knowledge nurses use to care for hospitalized patients. FINDINGS: Five discrete types of nursing knowledge that nurses use in practice emerged: personal practice knowledge, theoretical knowledge, procedural knowledge, ward cultural knowledge and reflexive knowledge. CONCLUSION: All three moments in the history of nursing knowledge were found to be concurrently present in nursing practice. Ward cultural knowledge and procedural knowledge reflect the rule-based descriptive knowledge of the first moment, theoretical knowledge and personal practice knowledge reflect the explanatory dualist knowledge of the second moment and reflexive knowledge reflects the critical and integrative knowledge of the third moment. PMID- 18426457 TI - Music improves sleep quality in students. AB - AIM: This paper is a report of a study to investigate the effects of music on sleep quality in young participants with poor sleep. BACKGROUND: Sleep disorders may result in fatigue, tiredness, depression and problems in daytime functioning. Music can reduce sympathetic nervous system activity, decrease anxiety, blood pressure, heart and respiratory rate and may have positive effects on sleep via muscle relaxation and distraction from thoughts. Control groups have not been used in most previous studies. METHODS: We used a three-group repeated measures design. Ninety-four students (aged between 19 and 28 years) with sleep complaints were studied in 2006. Participants listened for 45 minutes either to relaxing classical music (Group 1) or an audiobook (Group 2) at bedtime for 3 weeks. The control group (Group 3) received no intervention. Sleep quality was measured using the Pittsburg Sleep Quality Index before the study and weekly during the intervention. Depressive symptoms in experimental group participants were measured using the Beck Depression Inventory. RESULTS: Repeated measures anova revealed a main effect of TIME (P < 0.0001) and an interaction between TIME and GROUPS (P < 0.0001). Post hoc tests with Bonferroni correction showed that music statistically significantly improved sleep quality (P < 0.0001). Sleep quality did not improve statistically significantly in the audiobook and the control group. Depressive symptoms decreased statistically significantly in the music group (P < 0.0001), but not in the group listening to audiobooks. CONCLUSION: Relaxing classical music is an effective intervention in reducing sleeping problems. Nurses could use this safe, cheap and easy to learn method to treat insomnia. PMID- 18426458 TI - Factors influencing osteoporosis preventive behaviours: testing a path model. AB - AIM: This paper is a report of a study to test a model of certain factors influencing people engaging in osteoporosis preventive behaviours, and to estimate the direct and indirect effects of personal and social factors on whether people engage in those behaviours. BACKGROUND: Osteoporosis preventive behaviours, including exercise and calcium intake, help decreasing the risk of developing osteoporosis. Reasons for engaging in osteoporosis preventive behaviours are complex and influenced by personal and social factors. Years of education, self-efficacy, knowledge of osteoporosis, social support and social capital have been indicated to increase people engaging in osteoporosis preventive behaviours; but age has been shown to decrease those behaviours. METHODS: The proposed model was developed using Social Cognitive Theory and a conceptual framework for addressing the social context of health behaviour. A correlational cross-sectional study was carried out in 2005, using questionnaires and a convenience sample of 243 participants. Data analysis included descriptive statistics, correlation techniques and path analysis. RESULTS: Self-efficacy was a better predictor of engaging in osteoporosis preventive behaviours than were the other variables. Social capital had a statistically significant direct and indirect effect on osteoporosis preventive behaviours. The modified path model showed good fit with the data. CONCLUSION: The associations between personal and social factors extend our knowledge from previous studies and increase our understanding of the complex relationships among the study variables. The model provides guidance for future nursing practice, research, and education programs related to osteoporosis prevention. PMID- 18426459 TI - Night duty as an opportunity for learning. AB - AIM: This paper is a report of a study to examine what opportunities night nurses have to learn in terms of being able to distinguish variations in the patients' conditions. BACKGROUND: Night nurses often lack access to the formalized in service training offered to day nurses. As every clinical experience can be seen as an opportunity for learning, learning takes place even at night. However, the learning of night nurses has not been studied previously. METHOD: This study is based on interviews with a convenience sample of 10 night nurses at a medium sized Swedish hospital in 2001. These interviews were reanalysed in 2006 concerning learning situations. The interviews were tape-recorded, transcribed verbatim, coded and examined using latent content analysis. FINDINGS: There are certain opportunities for learning during the night shift, and three learning situations come to the fore: (1) the report situation, (2) the personal assessment round, where the nurses form their own picture of the patient, (3) in assessment prior to contact with the doctor on duty. Nurses learn from variations in patients' conditions and when they have to report their experience verbally. Learning does take place at night and gestalt psychology is a helpful tool for understanding how former knowledge and experience affect night nurses' learning. CONCLUSION: Knowledge developed during the night shift is a neglected field. There is a need for further investigations of what night nurses learn, and this knowledge ought to be integrated in the body of nursing knowledge. PMID- 18426460 TI - Re-examination of risk factors for non-Clostridium difficile-associated diarrhoea in hospitalized patients. AB - AIM: This paper is a report of a study to determine the incidence of non Clostridium difficile-associated diarrhoea in hospitalized patients and to re evaluate clinical characteristics and other risk factors related to non-C. difficile-associated diarrhoea. BACKGROUND: Numerous factors are thought to be responsible for diarrhoea in hospitalized patients. Reports about the diarrhoeal effects of some medications administered concomitantly with tube feeding have stimulated reappraisal of the influence of tube feeding as a potential cause. METHOD: This study was a secondary analysis of data of 154 hospitalized patients collected during a prospective epidemiological study from 1992 to 1993. The secondary analysis was completed in 2006 in order to investigate unanswered questions of current importance. FINDINGS: The sample was predominantly male and middle aged; approximately 50% were tube fed, and 25% were in an intensive care unit. The incidence of diarrhoea was 35%. Increased severity of illness as well as the combination of sorbitol-containing medication administration and tube feeding were found to be statistically significant factors in the development of diarrhoea. CONCLUSION: As diarrhoea in hospitalized patients appears to be multifactorial, use of an algorithm to systematically evaluate and manage related factors is recommended. PMID- 18426461 TI - Ethical issues in research with homeless youths. AB - AIM: This paper is a report of a study to document researcher, healthcare provider and programme administrators' experiences with ethical issues in research with homeless youths in North America. BACKGROUND: While there are legal and ethical guidelines for research with adolescents and with vulnerable populations in general, there are no specific guidelines for the ethical conduct of research with homeless youths. METHODS: Using a web-based questionnaire, healthcare and social service providers, programme administrators and researchers working with homeless young people throughout the United States of America and Canada were surveyed in 2005. The survey group consisted of 120 individuals; a total of 72 individuals completed the survey. Survey questions included experiences with using incentives in research with homeless youths, consent and experiences with ethics review boards. Numerical data were analysed using frequencies and cross-tabulations. Text data were analysed qualitatively. FINDINGS: Researchers doing mental health and/or substance use research tended to use money as a research incentive, whereas healthcare providers and programme administrators tended to use non-monetary incentives. The majority of respondents reported using written consent for research from homeless youths, including minors. Respondents reporting difficulties with ethics review boards were mainly involved with intervention research. CONCLUSION: Consensus is needed from a variety of stakeholders, including homeless youths and service providers, on use of various types of research incentives for different types of research, as well as use of consent for homeless youths who are minors. PMID- 18426462 TI - Research on couples: are feminist approaches useful? AB - AIM: This paper is an exploration of the utility and value of feminist approaches when undertaking narrative-based research with partner dyads (within both heterosexual and same sex partnerships) and when researching sensitive issues. BACKGROUND: Adverse life events or conditions experienced by individuals have been found also to have a negative impact on their partners. Most literature addressing partner issues uses quantitative methods, and existing qualitative research on couples has traditionally interviewed only one person in the partnership or coupled partners together. There is little discussion in the literature about the use of feminist research when researching male perspectives and experiences, and even less discussion of the possibilities that feminist research methods bring to the study of couple dyads. DISCUSSION: Qualitative methodologies informed by feminist perspectives, including issues of reciprocity and self disclosure, can be used to unpack structural, personal and political issues related to couples' experiences. A feminist approach allows us to show that the origin of oppression is not personal but very much about power and that men as well as women, regardless of their sexuality, may experience the effects of oppression. Narrative and story-telling complements feminist research because of the value it assigns to the storytellers. CONCLUSION: To care for women effectively, we must also consider the experiences of their partners as the health of one partner has the potential to impact on the other. The concept of oppression is not absent, but indeed is illuminated, in the lives of some men. Gathering stories using feminist perspectives enhances respect and mutuality in the research process. PMID- 18426463 TI - Concept synthesis of the art of nursing. AB - AIM: The aim of this paper is to add clarity to the concept art of nursing. BACKGROUND: Nurses have attempted to explicate the art of nursing since the time of Florence Nightingale. Despite these efforts, the concept remains obscure. A clearer conceptualization of nursing art is needed to help guide practice and research. METHOD: Fifty-nine English-language narratives (e.g. editorials, letters, anecdotes, brief items, etc.) that discuss the art of nursing were located using the Cumulative Index to Nursing & Allied Health Literature (1982 2006). Qualitative analysis and synthesis of these documents was completed using an adaptation of Walker and Avant's concept synthesis method. FINDINGS: The art of nursing consists of expert use and adaptation of empirical and metaphysical knowledge and values. It involves sensitively adapting care to meet the needs of individual patients, and in the face of uncertainty, the discretionary use of creativity. The art of nursing provides for enhanced mental and physical well being among patients. It also results in professional satisfaction and personal growth among nurses. CONCLUSION: Results from this investigation substantiate existing knowledge related to the art of nursing. In addition, the findings extend that knowledge by more clearly identifying the antecedents, attributes and outcomes of nursing art. Specifically, the epistemological and ontological components are more clearly identified. Clarity of this type is important in terms of future development of discipline-specific practice and research. PMID- 18426464 TI - In response to Foster J., Bidewell J., Buckmaster A., Lees S. & Henderson-Smart D. (2008) Parental stress and satisfaction in the non-tertiary special care nursery. Journal of Advanced Nursing61(5), 522-530. PMID- 18426465 TI - Information sharing, knowledge transfer and patient safety. PMID- 18426466 TI - Re: Knowledge transfer applied to the Australian health care system. PMID- 18426467 TI - Spiritual assessment in intensive and cardiac care nursing. AB - AIM: The aim of this paper is to explore various approaches to spiritual assessments in contemporary intensive and cardiac care (ICU/CCU) environments. BACKGROUND: Despite the increasing recognition that spiritual care is essential for quality patient care, an agreed spiritual assessment approach and tool for use in ICU/CCU settings remains elusive. METHOD: An overview of spiritual assessment and spiritual assessment tools. CONCLUSION: It is suggested that the staff in ICU/CCU nursing settings choose or develop a formal assessment tool that most closely matches their considered collective definition of spirituality, which has been considered in light of their mission statement and philosophy of care. RELEVANCE TO CLINICAL PRACTICE: Spiritual assessment is essential to formulate a care plan as spiritual care provides a powerful inner resource to critically ill patients in acute clinical environments. PMID- 18426468 TI - What factors influence arterial blood gas sampling patterns? AB - STUDY OBJECTIVE: To investigate if patterns of arterial blood gas (ABG) sampling were influenced by values of fractional inspiratory oxygen (FiO2), partial pressure of carbon dioxide (PCO2), partial pressure of oxygen (PO2) and oxygen saturation (%SaO2). SETTING: An intensive care unit (ICU) in a university teaching hospital located in the North of England, UK. DESIGN: A retrospective, descriptive, correlation study based on patient records. PARTICIPANTS: All patients admitted to the ICU for 24 hours or greater and who had an arterial line in situ. MEASUREMENTS AND RESULTS: The study included the records of 65 patients consecutively admitted to the ICU. Patients in this study had more blood gases taken than reported elsewhere in the literature. While consistent correlation was found between values of FiO2, PCO2, PO2 and %SaO(2), values of PO2 were the most consistent. CONCLUSIONS: Values of PO2 are associated with frequency of ABG sampling and to a lesser extent on FiO2. Nurses in this study opted to track changes in oxygenation using ABGs despite continuous monitoring of %SaO2. PMID- 18426469 TI - Therapeutic relationships in critical care nursing: a reflection on practice. AB - AIMS: The aim of this paper is to gain a greater understanding of the emotional intelligence required to form therapeutic nurse-patient relationships in the context of critical care nursing. CONTEXT: There is currently a demand for patient-centred nursing in all aspects of health care to improve both quality of care and consumer satisfaction. Central to patient-centred care is the presence of a therapeutic nurse-patient relationships, but the development of these relationships present a challenge in the acute and technological world of critical care nursing. THEORETICAL FRAMEWORK: Using Titchen's Skilled Companionship Model as a guide as well as empirical and theoretical knowledge on nurse-patient relationships, this paper presents a reflection on a relationship that the author developed with a patient and his family encountered in her practice as a critical care nurse. REFLECTIVE CONCLUSIONS: Engaging in therapeutic nurse-patient relationships can expose nurses to emotional pain in the context of critical care nursing. The process of reflection can facilitate critical care nurses in the development of the emotional intelligence required to develop and maintain these relationships and this enhances the care of critically ill patients and their families. PMID- 18426470 TI - Therapeutic hypothermia following cardiac arrest: a review of the evidence. AB - AIMS AND OBJECTIVES: This paper aims to undertake a review on the current evidence available on therapeutic hypothermia (TH) following cardiac arrest. BACKGROUND: The use of TH has been associated as a potential treatment for a number of medical conditions including head injury and cerebral vascular accidents. Within the past decade, there have been numerous studies focusing upon the use of hypothermia following cardiac arrest. This paper evaluates the research on the use of TH following cardiac arrest and provides recommendations for clinical practice. Evidence from randomized controlled trials that are reviewed in this paper found that neurological outcome and mortality were significantly improved following inducing hypothermia following cardiac arrest. SEARCH STRATEGIES: The following databases were accessed: Bandolier, Embase, Medline, Science Direct, CINAHL, Blackwell Synergy, Nursing Collection, Cochrane Central Register of Controlled Trials, Cochrane Database of Systematic Reviews and the National Electronic Library for Health. The following key words were used to search the databases: 'Therapeutic hypothermia', 'Induced hypothermia', 'cooling post cardiac arrest' and 'post cardiac arrest care'. INCLUSION AND EXCLUSION CRITERIA: Only evidences published within the past 10 years and written in English were included. Studies on TH for the treatment of raised intracranial pressure were excluded. CONCLUSIONS: All adult patients who have return of spontaneous circulation and remain unconscious following cardiac arrest should be considered for TH between 32 degrees C and 34 degrees C for at least 12-24 h as this will improve patient mortality and morbidity. Acute hospitals need to devise policies and guidelines on the use of TH following cardiac arrest that include methods on how to achieve effective cooling by cold i.v. infusions, ice packs or purchasing specific cooling mattresses. PMID- 18426471 TI - End-of-life care in UK critical care units--a literature review. AB - AIM: To appraise literature concerning end-of-life care (ELC) in adult critical care units in the UK in order to improve clinical practice. OBJECTIVE: To understand the interplay between legal and ethical, political, societal aspects of ELC for sustainable quality care. BACKGROUND: Significant changes in health care policy for the critically ill patient have occurred since 1999. Simultaneously, the government is committed to improving care for the dying by integrating the palliative care ethos across the National Institutes of Health (NHS) to include non-cancer sufferers. Death continues to be a feature of critical illness, particularly following the decision to withhold/withdraw life prolonging treatments. SEARCH STRATEGY: A search of MEDLINE, BNI, CINAHL and PSYCinfo using key words revealed very few results; consequently, the search was broadened to include ASSIA, King's Fund, TRIP, Healthstar, NHS Economic Evaluation Data, Cochrane, professional journals and government documents. CONCLUSIONS: The literature reveals a paradigm shift from critical to palliative care, in other words, from a reductionist approach to a more humanistic approach in the acute setting. When treatment is deemed futile, quality ELC involving the assessment, ongoing assessment and care after death becomes the new goal for the critical care team. To practice ELC competently, nurses require organizational and educational support at local and national levels. RELEVANCE TO CLINICAL PRACTICE: Although medico-legal decision-making is not part of their professional role, critical care nurses have an extraordinary opportunity to make a difference to the dying patient and their family and their acceptance of death. PMID- 18426472 TI - A dedicated retrieval and transfer service: the QUARTS Project. AB - BACKGROUND: The transfer and/or retrieval of a critically patient is inherently dangerous not only for the patient but for staff as well. The quality and experience of unplanned transfers can influence patient mortality and morbidity. However, international evidence suggests that dedicated transfer/retrieval teams can improve mortality and morbidity outcomes. AIMS: The initial aim of this paper is to describe an in-house competency-based training programme, which encompasses the STaR approach to develop members of our existing nursing team to be part of the dedicated transfer/retrieval service. The paper also presents audit data findings which examined the source of referrals, number of patients actually transferred and clinical status of those being transferred. RESULTS: Audit data illustrate that the most frequent source of referrals comes from Accident and Emergency and the Surgical Directorate with the most common presenting condition being cardio-respiratory failure or arrest. Audit data reveal that the number of patients actually transferred or retrieved is relatively small (33%) compared with the overall number of requests for assistance. However, 36% of those patients transferred had a level 2 or level 3 acuity status that necessitated the admission to a critical care area. CONCLUSIONS: A number of studies have concluded that the ill-experienced and ill-equipped transfer team can place patients' at serious risk of harm. Whether planned or unplanned, dedicated critical care transfer/retrieval teams have been shown to reduce patient mortality and morbidity. PMID- 18426473 TI - Evaluating evidence-based practice within critical care. AB - BACKGROUND AND AIMS: Between 2002-2005 the Trust undertook an action research project to evaluate a corporate practice development strategy. During this period clinicians became practitioner-researchers utilising a variety of methods to evaluate the influence of practice development. One aspect of this focused upon evaluation of evidence based guidelines. This article concentrates upon this process and the learning from this within critical care. METHOD: Within critical care it was recognised that the standard of guidelines and protocols varied in terms of the amount of evidence used to underpin decision making. A group was set up to evaluate and appraise these using a structured format such as the Appraisal of Guidelines for Research and Evaluation (AGREE) Instrument. FINDINGS: The initial evaluation (cycle 1) highlighted learning associated with the process of using the instrument within critical care, as well as where the quality of the guidelines could be improved. The second cycle of evaluation demonstrated that implementation of the action plans as a consequence of cycle 1 resulted in an improvement in the quality of the guidelines. It also resulted in streamlining the process of undertaking guideline appraisal across a Trust. DISCUSSION AND CONCLUSIONS: Action resulting from analysis of the findings of cycle 1 led to a cultural change in which the structure of a tool such as the AGREE instrument could be beneficial in the development of future guidelines. This has been sustained both within critical care and Trust wide with various initiatives such as the establishment of critical care multidisciplinary guideline development groups and a Trust wide electronic library management system. PMID- 18426474 TI - Commentary: Nemoto T et al. (1999). Automatic control of pressure support mechanical ventilation using fuzzy logic. PMID- 18426475 TI - Phytochrome-driven changes in respiratory electron transport partitioning in soybean (Glycine max. L.) cotyledons. AB - After it was observed that light induces changes in electron partitioning between the cytochrome and the alternative pathway, the focus interest was directed to assessing what type of photoreceptors are involved and the extent of such modifications. Studies on 5-day-old soybean (Glycine max L.) cotyledons using an oxygen isotope fractionation technique showed that phytochrome is involved in changes in electron partitioning between the cytochrome and the alternative respiratory pathway. A follow-up of a previous study, showing that 5 min of white light caused changes in mitochondrial electron partitioning, demonstrated that while blue light was not involved in any such changes, red light caused a significant shift of electrons toward the alternative pathway. The major shift, observed after 24 h of light, is mainly due to both a decrease in the activity of the cytochrome pathway and an increase in the activity of the alternative pathway. The involvement of a phytochrome receptor was confirmed by demonstration of reversibility by far-red light. The implications of the possible involvement of phytochrome in the regulation of mitochondrial electron transport are discussed. PMID- 18426476 TI - Short- and long-term modulation of the lutein epoxide and violaxanthin cycles in two species of the Lauraceae: sweet bay laurel (Laurus nobilis L.) and avocado (Persea americana Mill.). AB - Short- and long-term responses of the violaxanthin (V) and lutein epoxide (Lx) cycles were studied in two species of Lauraceae: sweet bay laurel (Laurus nobilis L.) and avocado (Persea americana L.). The Lx content exceeded the V content in shade leaves of both species. Both Lx and V were de-epoxidised on illumination, but only V was fully restored by epoxidation in low light. Violaxanthin was preferentially de-epoxidised in low light in L. nobilis. This suggests that Lx accumulates with leaf ageing, partly because its conversion to lutein is limited in shade. After exposure to strong light, shade leaves of avocado readjusted the total pools of alpha- and beta-xanthophyll cycles by de novo synthesis of antheraxanthin, zeaxanthin and lutein. This occurred in parallel with a sustained depression of F(v)/F(m). In Persea indica, a closely related but low Lx species, F(v)/F(m) recovered faster after a similar light treatment, suggesting the involvement of the Lx cycle in sustained energy dissipation. Furthermore, the seasonal correlation between non-reversible Lx and V photoconversions and pre dawn F(v)/F(m) in sun leaves of sweet bay supported the conclusion that the Lx cycle is involved in a slowly reversible downregulation of photosynthesis analogous to the V cycle. PMID- 18426477 TI - The SUI-homologous translation initiation factor eIF-1 is involved in regulation of ion homeostasis in rice. AB - Halophytes survive high salinity by using complex adaptive mechanisms. In a search for novel molecular mechanisms involved in salt acclimation, transcript analyses revealed increased expression of a SUI-homologous translation initiation factor eIF-1 in the salt-tolerant grass species Festuca rubra ssp. littoralis but not in rice. Upon analysis of the cell specificity of eIF-1 transcription by in situ polymerase chain reaction (PCR), predominant signals were detected in rice leaf mesophyll. To further examine the role of eIF-1 in salt tolerance, transgenic rice plants were generated that over-express this factor under the control of the CaMV-35S promoter. The eIF-1 over-expressing lines showed improved growth under salt stress that was correlated with maintenance of photosynthetic activity and reduced Na(+) and Cl(-) accumulation in leaves. The transgenic rice lines also activated expression of the vacuolar H(+)-ATPase. In addition, an oxidoreductase that belongs to the aldo/keto reductase family was identified as a gene with modified expression in the eIF-1 over-expressing lines, compared with wild-type rice. Our data suggest that eIF-1 has a central function in salt-stress adaptation in rice by regulating ion accumulation and the intracellular redox status. PMID- 18426478 TI - Characterization of resistance gene analogs with a nucleotide binding site isolated from a triploid white poplar. AB - The majority of cloned plant disease resistance genes (R genes) encode a nucleotide binding site (NBS) and a leucine-rich repeat (LRR) domain. In this study, to better understand the R genes in white poplar, 59 resistance gene analogues (RGAs) were identified from a triploid white poplar [(Populus tomentosa x Populus bolleana) x P. tomentosa], based on conserved NBS regions. The 59 RGAs were phylogenetically classified into 10 subfamilies, and 54 RGAs with open reading frames (ORFs) were further grouped into two classes, toll and interleukin 1 receptor (TIR) and non-TIR. BLAST searches with reference to the genomic sequence of Populus trichocarpa found 96 highly homologous regions distributed in 37 loci, suggesting the abundance and divergence of NBS-encoding genes in the triploid poplar genome. Within subfamilies 1-3, the average non synonymous/synonymous substitution (omega) rates were < 1, indicating purifying selection on these RGAs, but some sites were clearly under diversifying selection with omega > 1. Many intergenic exchanges were also detected among these RGAs, indicating a probable role in homogenising NBS domains. Quantitative real-time PCR analysis revealed dramatic variations in the transcript level of 18 RGAs in the mature leaves, bark and roots of the triploid poplar, and identified two RGAs that had significantly higher level of transcripts in bark, four RGAs in mature leaves, and 14 in the above-ground portion of poplars, suggesting their probable roles in resistance against diseases attacking the organs. Our results shed light on genetic resources of poplar resistance and will be useful for further resistance gene isolation and exploitation. PMID- 18426479 TI - Effects of overexpression of endogenous phenylalanine ammonia-lyase (PALrs1) on accumulation of salidroside in Rhodiola sachalinensis. AB - Salidroside, a novel effective adaptogenic drug extracted from the medicinal plant Rhodiola sachalinensis A. Bor, can be derived from phenylalanine or tyrosine. Due to the scarcity of R. sachalinensis and its low yield of salidroside, there is great interest in enhancing production of salidroside by the plant. In this study, a cDNA clone encoding phenylalanine ammonia-lyase (PAL) was isolated from R. sachalinensis using rapid amplification of cDNA ends. The resulting cDNA was designated PALrs1. It is 2407-bp long and encodes 710 deduced amino acid residues. Southern blot analysis of genomic DNA indicated that the PAL gene family is composed of three to five genes in the R. sachalinensis genome. Northern blot analysis revealed that transcripts of PALrs1 were present in calli, leaves and stems, but expression in roots was very low. The PALrs1 under the 35S promoter with double-enhancer sequences from CaMV-Omega and TMV-Omega fragments was transferred into R. sachalinensis via Agrobacterium tumefaciens. PCR and PCR Southern blot confirmed that the PALrs1 gene had been integrated into the genome of transgenic plants. Northern blot analysis revealed that the PALrs1 gene had been expressed at the transcriptional level. High-performance liquid chromatography indicated that overexpression of the PALrs1 gene resulted in a 3.3 fold increase in p-coumaric acid content, as expected. In contrast, levels of tyrosol and salidroside were 4.7-fold and 7.7-fold, respectively, lower in PALrs1 transgenic plants than in controls. Furthermore, overexpression of the PALrs1 gene resulted in a 2.6-fold decrease in tyrosine content. These data suggest that overexpression of the PALrs1 gene and accumulation of p-coumaric acid did not facilitate tyrosol biosynthesis; tyrosol, as a phenylethanoid derivative, is not derived from phenylalanine; and reduced availability of tyrosine most likely resulted in a large reduction in tyrosol biosynthesis and accumulation of salidroside. PMID- 18426480 TI - Bis(guanylhydrazones) negatively affect in vitro germination of kiwifruit pollen and alter the endogenous polyamine pool. AB - Bis(guanylhydrazones) are a class of compounds known to interfere with the metabolism of polyamines (PAs). Among them, the methylglyoxal derivative (MGBG) has been studied most thoroughly. Because PAs and their biosynthetic enzymes are strongly involved in pollen tube organization, emergence and elongation, a number of these inhibitors have been studied in the present work for their effects on the in vitro performance of kiwifruit (Actinidia deliciosa) pollen. Increasing concentrations of several bis(guanylhydrazones) in the range 0.05-1 mM were checked for their effect on pollen germination. Most of the compounds tested showed a dose-dependent inhibitory effect on tube emergence, which was established very early during incubation. At 0.5 mM, the methylpropylglyoxal derivative (MPGBG) had a stronger inhibitory effect than MGBG. To verify whether the inhibitors reached their metabolic target, PA levels and S-adenosylmethionine decarboxylase (SAMDC) activity were determined in pollen germinated in the presence or absence (controls) of 0.5 mM bis(guanylhydrazones). Spermidine (Spd) content was significantly reduced in the treated pollen, and this effect was more pronounced after treatment with MGBG than with MPGBG. An early and strong reduction in SAMDC activity was observed after exposure to either inhibitor. Inhibition of pollen germination by MGBG or MPGBG could not be reversed by the addition of exogenous Spd, which per se was inhibitory. Taken together, our results suggest that bis(guanylhydrazones) alter PA metabolism and negatively affect kiwifruit pollen germination, even though a strict cause-effect relationship could not be established, and other mechanisms, unrelated to PA activity, must be involved. PMID- 18426481 TI - Transcriptional differences between the male-sterile mutant bcms and wild-type Brassica campestris ssp. chinensis reveal genes related to pollen development. AB - A novel male-sterile mutant which lacks mature pollen, Brassisa campestris male sterile (bcms), was identified in Brassica campestris L. ssp. chinensis Makino (syn. B. rapa ssp. chinensis). Genetic analysis revealed that bcms was controlled by a single recessive mutation locus. Genome-wide transcriptional profiling was performed on the flower buds of both the bcms mutant and the wild-type from which it originated, and profiling analysis indicated that there were numerous changes in gene expression attributable to the gene mutation. This mutation resulted in down-regulation of a variety of genes and up-regulated expression of a few other genes. A total of 51 transcript-derived fragments (TDFs) were isolated: 32 specifically and 12 predominantly accumulated in wild-type flower buds, and two specifically and five predominantly accumulated in bcms flower buds. Sequence analysis showed that some of these TDFs share significant similarities with genes involved in different aspects of cellular development, such as signal transduction, cell wall biosynthesis and regulation. Most other TDFs showed no or very poor sequence similarities to entries in any database and might represent new candidate proteins involved in pollen development. Furthermore, spatial and temporal expression pattern analysis of 20 genes derived from cDNA-amplified fragment length polymorphism in different tissues of both the bcms and wild-type plants revealed their complex and dynamic expression patterns. The bcms mutant and the genes isolated in this paper provide excellent material for future studies on the molecular mechanism of male sterility. PMID- 18426482 TI - The role of gene flow in shaping genetic structures of the subtropical conifer species Araucaria angustifolia. AB - The morphological features of pollen and seed of Araucaria angustifolia have led to the proposal of limited gene dispersal for this species. We used nuclear microsatellite and AFLP markers to assess patterns of genetic variation in six natural populations at the intra- and inter-population level, and related our findings to gene dispersal in this species. Estimates of both fine-scale spatial genetic structure (SGS) and migration rate suggest relatively short-distance gene dispersal. However, gene dispersal differed among populations, and effects of more efficient dispersal within population were observed in at least one stand. In addition, even though some seed dispersal may be aggregated in this principally barochorous species, reasonable secondary seed dispersal, presumably facilitated by animals, and overlap of seed shadows within populations is suggested. Overall, no correlation was observed between levels of SGS and inbreeding, density or age structure, except that a higher level of SGS was revealed for the population with a higher number of juvenile individuals. A low estimate for the number of migrants per generation between two neighbouring populations implies limited gene flow. We expect that stepping-stone pollen flow may have contributed to low genetic differentiation among populations observed in a previous survey. Thus, strategies for maintenance of gene flow among remnant populations should be considered in order to avoid degrading effects of population fragmentation on the evolution of A. angustifolia. PMID- 18426483 TI - Daylength responses of various morphogenetic events after panicle initiation in late japonica rice (Oryza sativa L. ssp. japonica). AB - Plant morphogenesis is an intimate cooperation between environmental signals and a genetically-encoded developmental programme. Although all stages subsequent to floral induction may be influenced by daylength, the information available has come from diverse species during different morphogenetic events. This makes it difficult to investigate how photoperiodic signalling is integrated into various steps of the developmental programme. Here, we report that, in late japonica rice lines, morphogenetic events, including 12th leaf, axis of the main panicle, awn of the glum, anthers, elongation rate of internodes, and pollen fertility of the photoperiod-sensitive genic male-sterile (PGMS) rice, are significantly affected by daylength after panicle initiation. These data indicate that daylength affects many morphologic events, and that changes in morphology are mainly determined by the characteristics of the events themselves. These findings lay a foundation for future investigations into how potentially common photoperiodic signalling system(s) are integrated with diversified developmental events. In addition, we discussed the essential nature of PGMS rice. PMID- 18426484 TI - Age-specific, density-dependent and environment-based mortality of a short-lived perennial herb. AB - Density-independent and density-dependent processes affect plant mortality. Although less well understood, age-specific mortality can also play an important role in plant mortality. The goal of this study was to analyse several factors accounting for mortality in the Mediterranean short-lived perennial herb Lobularia maritima. We followed three cohorts of plants (from emergence to death) during 4 years in field conditions. We collected data on plant mortality of the effect of biotic agents (moth larvae and mycoplasma-like organisms, MLOs) and environmental variables. We also estimated density-dependent relationships affecting the fate of seedlings and adults. Results show that cohorts differed in their survival curves and ageing significantly increased mortality risk. Seedling mortality was density-dependent whereas adult mortality was not affected by density. MLO infection led to higher plant mortality whereas moth larvae attack did not affect plant mortality. In general, seedlings and adult plants experienced the highest mortality events in summer. We found, however, weak relationships between weather records and plant mortality. Age and size structures were not correlated. Overall, this study provides a comprehensive review of age-specific, density-dependent and density-independent factors that account for mortality of L. maritima plants throughout their life cycle in field conditions, highlighting the fact that age is an important factor in determining plant population dynamics. PMID- 18426485 TI - Underwater germination potential of common ash seed (Fraxinus excelsior L.) originating from flooded and non-flooded sites. AB - Oxygen availability is one of the main factors affecting seed germination capacity. On floodplains, oxygen supply is seasonally restricted because of flooding events. Seed ecotypes are able to cope with total submergence to different extents, depending on the species and its position in a given environmental gradient. This investigation tested three provenances of common ash (Fraxinus excelsior L.), one from a lowland site and two from upland sites, for germination capacity and germination velocity under water. The final germination capacity of all provenances was unaffected, but the germination velocity was decreased for all provenances. The adaptational significance of these results is discussed. PMID- 18426486 TI - Effects of ultraviolet radiation and temperature on the ultrastructure of zoospores of the brown macroalga Laminaria hyperborea. AB - The interactive effects of an 8 h exposure to UV radiation and altered temperatures on the ultrastructure and germination of zoospores of the sublittoral brown alga Laminaria hyperborea (Gunn.) Foslie were investigated for the first time. Spores were exposed to four temperatures (2, 7, 12 and 17 degrees C) and three light regimes (PAR, PAR + UV-A, PAR + UV-A+UV-B). Freshly-released spores of L. hyperborea lack a cell wall and contain a nucleus with fine granular nucleoplasm and a nucleolus, one chloroplast, several mitochondria, dictyosomes and an endoplasmatic reticulum. Further, several kinds of so-called adhesive vesicles, lipid globuli and physodes containing UV-absorbing phlorotannins are embedded in the cytoplasm. No eye-spot is present. Physodes were found but they were rare and small. After an 8 h exposure to UV-B, the nucleoplasm had a mottled structure, chloroplasts contained plastoglobuli, the structure of the mitochondria changed from crista- to sacculus-type and germination was strongly inhibited at all temperatures. UV-A only had an impact on the ultrastructure at the highest temperature tested. The strongest effects were found at 17 degrees C, where germination was reduced to 35%, 32% and 9% after exposure to PAR, PAR+UV-A and PAR + UV-A + UV-B, respectively. This study indicates that UV-B radiation has strong damaging effects on the physiology and ultrastructure of zoospores of L. hyperborea. The results are important for developing scenarios for the effect of enhanced UV radiation and increasing temperatures caused by global climate changes. PMID- 18426487 TI - A possible role of prolyl oligopeptidase during Linum usitatissimum (flax) seed development. AB - Involvement of prolyl oligopeptidases (POPs) in the control of several mammalian peptide hormone signalling pathways has been studied extensively in recent years. POPs are ubiquitous enzymes, but little attention has been paid to understanding their function in plants. Using a cDNA-AFLP approach, two flax (Linum usitatissimum) POP ESTs were identified as being specifically expressed in the early stages of flax seed development. This specific expression was confirmed using real time RT-PCR and in situ hybridisation approaches. Seed expression of Arabidopsis POP genes was measured and showed no specificity. Comparison between results obtained with flax and Arabidopsis is discussed in order to address a hypothetic function for POPs during seed formation. These results provide the first insights into POP gene expression and hypothetical function in plants. PMID- 18426489 TI - Do consecutive flower visits within a crown diminish fruit set in mass-flowering Hancornia speciosa (Apocynaceae)? AB - Hancornia speciosa is a self-incompatible, mass-flowering, sphingophilous fruit crop (mangaba) of northeast and central Brazil. The flowers have a precise pollination apparatus, which optimizes pollen transfer between flower and pollinator. While the pollination mechanism avoids self-pollination, mass flowering promotes geitonogamy. During a flower visit, almost half of the exogenous pollen grains adhering to the proboscis are deposited on the stigma surface. A pollination experiment with a nylon thread simulating six consecutive flower visits within a crown revealed that only the first two flowers visited (positions 1 and 2) are highly likely to set fruit. Super-production of flowers, and consequently obligate low fruit set, seem to be part of the reproductive strategy of the obligate outcrossing plant, Hancornia speciosa. PMID- 18426488 TI - Antioxidative enzymes and the Russian wheat aphid (Diuraphis noxia) resistance response in wheat (Triticum aestivum). AB - A crucial function of antioxidative enzymes is to remove excess reactive oxygen species (ROS), which can be toxic to plant cells. The effect of Russian wheat aphid (RWA), Diuraphis noxia (Mordvilko), infestation on the activities of antioxidative enzymes was investigated in the resistant (cv. Tugela DN) and the near-isogenic susceptible (cv. Tugela) wheat (Triticum aestivum L.). RWA infestation significantly induced the activity of superoxide dismutase, glutathione reductase and ascorbate peroxidase to higher levels in the resistant than in susceptible plants. These findings suggest the involvement of antioxidative enzymes in the RWA-wheat resistance response, which was accompanied by an early oxidative burst. The results are consistent with the role of ROS in the resistance response and the control of their levels to minimise toxic effects. PMID- 18426491 TI - Here, there and everywhere: the endocannabinoid system. PMID- 18426492 TI - Endogenous cannabinoids: structure and metabolism. AB - The finding of specific binding sites for Delta(9)-tetrahydrocannabinol, the psychoactive component of Cannabis sativa, has led to the discovery of the endocannabinoid system and has emphasised the physiological and pathological relevance of endocannabidoid lipid signalling. Subsequently, an increasing number of papers have been published on the biochemistry and pharmacology of endocannabinoids. An overview of the current understanding of structure and metabolism of the best studied endocannabinoids is provided, with a focus on the mechanisms responsible for their biosynthesis and inactivation. PMID- 18426493 TI - Cannabinoid receptors: where they are and what they do. AB - The endocannabinoid system consists of the endogenous cannabinoids (endocannabinoids), cannabinoid receptors and the enzymes that synthesise and degrade endocannabinoids. Many of the effects of cannabinoids and endocannabinoids are mediated by two G protein-coupled receptors (GPCRs), CB(1) and CB(2), although additional receptors may be involved. CB(1) receptors are present in very high levels in several brain regions and in lower amounts in a more widespread fashion. These receptors mediate many of the psychoactive effects of cannabinoids. CB(2) receptors have a more restricted distribution, being found in a number of immune cells and in a few neurones. Both CB(1) and CB(2) couple primarily to inhibitory G proteins and are subject to the same pharmacological influences as other GPCRs. Thus, partial agonism, functional selectivity and inverse agonism all play important roles in determining the cellular response to specific cannabinoid receptor ligands. PMID- 18426494 TI - Endocannabinoids and the brain immune system: new neurones at the horizon? AB - Whereas, in most brain compartments, neuronal cell renewal during early life is replaced by synaptic plasticity and the potentiation of existing pathways and connections, neurogenesis in the hippocampus occurs throughout adulthood. Neuronal progenitor cells in the dentate gyrus of the hippocampus are thought to be the gatekeepers of memory. Neural progenitor cell proliferation and differentiation depends on their intrinsic properties and local environment and is down-regulated in conditions associated with brain inflammation. Conversely, newly-formed neurones can survive despite chronic inflammation and, moreover, specifically arise within an inflammatory environment. Since the endocannabinoid system controls immune responses via multiple cellular and molecular targets and influences cell proliferation, fate decision and cell survival in the central nervous system, we summarise how neurogenesis might be regulated by brain cannabinoids, either directly or indirectly via the immune system. This review presents clear evidence that the cannabinoid system influences adult neurogenesis. However, there is considerable variability with regard to the strain, model and methods utilised and therefore it is difficult to compare studies investigating the cannabinoid system. As a result, it remains far from clear exactly how endocannabinoids regulate neurogenesis. PMID- 18426495 TI - Cannabinomimetic control of mast cell mediator release: new perspective in chronic inflammation. AB - The present review aims to elucidate the emerging role played by cannabinomimetic compounds in the control of mast cell activation. Mast cells are immune competent cells strategically localised at the sites directly interfacing with the external environment, which, in case of injury, regulate the immune response by the release of a plethora of both pre-formed and newly-synthesised mediators. However, although the main goal of mast cell activation is to initiate the inflammatory reaction, and thus maintain internal homeostasis, the consequences of dysregulated mast cell activation could be to chronically activate the inflammatory response as occurs in arthritis, inflammatory bowel diseases, atherosclerosis and asthma. Therefore, much effort has been made to develop compounds that act to prevent mast cell degranulation. Cannabinomimetic compounds (i.e. agents able to modulate endocannabinoid function) are considered as an emerging class of regulators of mast cell behaviour. We focus on the evidence for a cannabinomimetic control of both acute and chronic inflammatory disease, recognising a common mast cell origin for problems such as dermatitis, inflammatory gastrointestinal syndrome and granuloma formation. Special emphasis is provided for the recent promising results obtained with palmitoylethanolamide in human studies. In the light of evidence suggesting that the control of mast cell activation at an early time during an inflammatory process may account for its resolution, it is reasonable to propose that cannabinomimetic compounds, including palmitoylethanolamide and its congeners, could represent possible candidates for treating several chronic inflammatory diseases. PMID- 18426496 TI - A saturated N-acylethanolamine other than N-palmitoyl ethanolamine with anti inflammatory properties: a neglected story... AB - N-acylethanolamines, which include the endocannabinoid anandamide and the cannabinoid receptor-inactive saturated compounds N-palmitoyl ethanolamine and N stearoyl ethanolamine, are ethanolamines of long-chain fatty acids degraded by fatty acid amide hydrolase (FAAH) known to accumulate in degenerating tissues and cells. Whilst much evidence supports a protective anti-inflammatory role of both anandamide and N-palmitoyl ethanolamine, very little information is available with regard to the bioactivity of N-stearoyl ethanolamine. Employing a murine model of passive IgE-induced cutaneous anaphylaxis, we have found that N-stearoyl ethanolamine is endowed with marked anti-inflammatory properties in vivo, supporting the hypothesis that endogenous N-stearoyl ethanolamine is, in analogy to N-palmitoyl ethanolamine, a bioactive signalling lipid capable of downregulating allergic inflammation in the skin. This effect, although mimicked by synthetic, non-selective, CB(1)/CB(2) receptor agonists, such as WIN55, 212-2, was not sensitive to CB(1) or CB(2) receptor antagonists, but rather was fully reversed by capsazepine, a competitive antagonist of the TRPV1 receptor. Moreover, CB(1) receptor antagonists, although effective in antagonising the WIN55,212-2-induced hypothermia, did not reduce the anti-inflammatory effect of WIN55,212-2, whilst CB(2) receptor antagonists, per se inactive, potentiated the WIN55,212-2 effect, suggesting an involvement of non-CB(1)/CB(2) receptors in the anti-inflammatory action of WIN55,212-2. All this, together with demonstration of FAAH as a major regulator of the in vivo concentrations of saturated N-stearoyl ethanolamine, in addition to N-palmitoyl ethanolamine, raise the speculation that pharmacological treatments with saturated N-acylethanolamines such as N-stearoyl ethanolamine, or alternatively FAAH inhibitors able to increase their local concentration, rather than selective CB receptor agonists, might be of promising therapeutic benefit in reducing allergic inflammation in the skin. PMID- 18426497 TI - The role of the endocannabinoid system in the regulation of hypothalamic pituitary-adrenal axis activity. AB - The endocannabinoid system (ECS) is a recently identified neuromodulatory system, which is involved in several physiological processes and in disease. For example, the ECS not only represents the biological substrate of marijuana's effects, but also is known to modulate several neuroendocrine axes, including the hypothalamic pituitary-adrenal (HPA) axis. Although previous pharmacological studies using plant-derived or synthetic cannabinoids have implied a stimulating action on the HPA axis, more recent findings have led to the conclusion that an endogenous cannabinoid tone might exist, which is actually inhibiting the release of both adrenocorticotrophic hormone and glucocorticoids. Studies using mice lacking cannabinoid receptor CB(1) have demonstrated that presence and activity of these receptors is essential for the regulation of HPA axis activity. Interestingly, the effects of endocannabinoids on the HPA axis are consistent with their neuromodulatory action on brain neurotransmitter systems. Endocannabinoids have been found to mediate the nongenomic glucocorticoid-induced inhibition of the release of corticotrophin-releasing factor within the paraventricular nucleus of the hypothalamus. Altogether, these observations suggest that alterations of the endocannabinoid tone might be associated with the development of stress-related diseases, including anxiety, depression and obesity. PMID- 18426498 TI - Gastrointestinal regulation of food intake: general aspects and focus on anandamide and oleoylethanolamide. AB - The gastrointestinal tract plays a pivotal role in the regulation of food intake and energy balance. Signals from the gastrointestinal tract generally function to limit ingestion in the interest of efficient digestion. These signals may be released into the bloodstream or may activate afferent neurones that carry information to the brain and its cognitive centres, which regulates food intake. The rate at which nutrients become systemically available is also influenced by gastrointestinal motility: a delay in gastric emptying may evoke a satiety effect. Recent evidence suggests that the endocannabinoid anandamide and the related acylethanolamide oleoylethanolamide are produced in the intestine and might regulate feeding behaviour by engaging sensory afferent neurones that converge information to specific areas of the brain. The intestinal levels of these acylethanolamides are inversely correlated to feeding, as food deprivation increases intestinal levels of anandamide (which acts in the gut as a 'hunger signal'), while it decreases the levels of oleoylethanolamide (which acts in the gut as a 'satiety signal'). Additionally, these acylethanolamides, whose gastric levels change in response to diet-induced obesity, alter gastrointestinal motility, which might contribute to their effect on food intake and nutrient absorption. PMID- 18426499 TI - The endocannabinoid system and liver diseases. AB - Endogenous cannabinoids (EC) are ubiquitous lipid signalling molecules provided by a number of central and peripheral effects, which are mainly mediated by the specific cannabinoid receptors CB(1) and CB(2). Although the expression of these receptors is very low or even absent in the healthy liver, a considerable series of experimental studies and some clinical observations have recognised the EC system as an important player in the pathophysiology of liver diseases. The EC system is highly up-regulated during chronic liver diseases and, to date, it has been implicated in the pathogenesis of non-alcoholic fatty liver disease, progression of fibrosis to cirrhosis and the development of the cardiovascular abnormalities of cirrhosis, such as the hyperdynamic circulatory syndrome and cirrhotic cardiomiopathy. Furthermore, the EC system influences the mechanisms responsible for cell damage and the inflammatory response during acute liver injury, such as that resulting from ischaemia-reperfusion. Thus, molecules targeting the CB(1) and CB(2) receptors may represent potential therapeutic agents for the treatment of liver diseases. At present, the CB(1) antagonists represent the most attractive pharmaceutical tool to resolve fat accumulation in patients with non-alcoholic fatty liver disease and to treat patients with cirrhosis, as they may slow the progression of fibrosis and attenuate the cardiovascular alterations associated with the advanced stage of the disease. PMID- 18426500 TI - The role of the endocannabinoid system in atherosclerosis. AB - Our current understanding of the pathophysiology of atherosclerosis suggests a prominent role for immune responses from its initiation through its complications. Given the increasing prevalence of cardiovascular risk factors worldwide, there is an urgent need to better understand the underlying mechanisms to improve current treatment protocols. A growing body of evidence suggests that endocannabinoid signalling plays a critical role in the pathogenesis of atherogenesis and its clinical manifestations. Blocking CB(1) receptors has been shown to mediate not only weight reduction, but also several cardiometabolic effects in rodents and humans, indicating a potential relevance for the process of atherosclerosis. Activation of CB(2) receptors with Delta(9) tetrahydrocannabinol (THC) has been shown to inhibit atherosclerotic plaque progression in mice, mainly by inhibiting macrophage recruitment. Endocannabinoids released from endothelial cells, macrophages or platelets, reduce hypertension in rodents, a major risk factor for atherosclerosis. In addition, anandamide inhibits inflammatory gene expression in endothelial cells, and consequently monocyte adhesion. Conversely, endocannabinoids might also mediate pro-atherosclerotic effects by inducing platelet activation. In conclusion, the precise role of the endocannabinoid system during atherosclerosis is not yet understood. Whether increased endocannabinoid signalling is associated with disease progression and increased risk of acute thrombotic events remains to be determined. PMID- 18426501 TI - Endocannabinoids, blood pressure and the human heart. AB - The effects of exogenous and endogenous cannabinoids on the cardiovascular system have been the focus of extensive research. The direct and indirect effects of cannabinoids on heart and blood vessels depend upon experimental conditions, animal species, and, in humans, clinical background. Cannabinoids decrease blood pressure in hypertensive rodents primarily because of decrease cardiac contractility, leading researchers to postulate a role in the treatment of hypertension and cardiac hypertrophy. Rimonabant, the CB(1) receptor blocker in clinical use in many countries, induced a marked and sustained increase in cardiac contractility and blood pressure in hypertensive rats but, on the contrary, contributed to decrease blood pressure in weight-loss clinical trials especially in obese patients with hypertension. In the midst of the obesity pandemic and from the cardiometabolic point of view, the overactivation of the endocannabinoid system present in intra-abdominal obesity appears to be very harmful. Moreover, novel human findings suggest a relationship between CB(1) mediated overactive endocannabinoid system and nephrovascular damage. Overall, it appears that CB(1) blockade in obese patients behaves as a 'multiplier' of the many beneficial effects of body weight loss induced by a hypocaloric diet and increased physical activity (the 'lifestyle changes' that are so difficult to start and maintain). Thus, the concept - based mostly on experimental results using in vitro or animal models - that CB(1)-mediated endocannabinoid effects are beneficial for the cardiovascular system should be revised at least in obese patients. The results of long-term clinical trials such as the STRADIVARIUS and the CRESCENDO trials will tell whether the improvement in the cardiometabolic risk profile induced by Rimonabant translates into vascular changes, reducing the risk of myocardial infarction, stroke and cardiovascular death in patients with abdominal obesity. Time (and much more work) will tell us much more about cannabinoids and the human heart. PMID- 18426502 TI - Blood pressure lowering effects of rimonabant in obesity-related hypertension. AB - Obesity-related hypertension represents a common clinical condition characterised by complex pathophysiological and therapeutic features. From a pathophysiological view point, results of experimental and animal studies have led to the hypothesis that neurogenic mechanisms participate in the development and progression of the disease. The hypothesis is based on the evidence that metabolic (i.e. insulin resistance) and neural (sympathetic activation) alterations frequently co-exist in the obese hypertensive patient and that they reciprocally potentiate each other. From a therapeutic view point, the 2007 European Society of Hypertension/European Society of Cardiology emphasised the importance in this clinical condition of treatment not only through antihypertensive drugs but also via lifestyle changes and drug-induced interventions that reduce body weight. The four Rimonabant In Obesity (RIO) studies have shown that rimonabant can decrease body weight. A recent meta-analysis, based on the RIO results, showed that rimonabant, particularly in obese hypertensive patients, can also decrease - although modestly (2.8 mmHg for systolic and 2.2 mmHg for diastolic) - blood pressure. These effects, which appear to be triggered by the weight reduction induced by the drug, are clinically relevant because they contribute favourably to lower the elevated cardiovascular risk profile of the obese hypertensive patient. PMID- 18426503 TI - Endocannabinoids and the regulation of bone metabolism. AB - In mammals, including humans, bone metabolism is manifested as an ongoing modelling/remodelling process whereby the bone mineralised matrix is being continuously renewed. Recently, the main components of the endocannabinoid system have been reported in the skeleton. Osteoblasts, the bone forming cells, and other cells of the osteoblastic lineage, as well as osteoclasts, the bone resorbing cells, and their precursors, synthesise the endocannabinoids anandamide and 2-arachidonoylglycerol (2-AG). CB(1) cannabinoid receptors are present in sympathetic nerve terminals in close proximity to osteoblasts. Activation of these CB(1) receptors by elevated bone 2-AG levels communicates brain-to-bone signals as exemplified by traumatic brain injury-induced stimulation of bone formation. In this process, the retrograde CB(1) signalling inhibits noradrenaline release and alleviates the tonic sympathetic restrain of bone formation. CB(2) receptors are expressed by osteoblasts and osteoclasts. Their activation stimulates bone formation and suppresses bone resorption. CB(2) deficient mice display a markedly accelerated age-related bone loss. Ovariectomy induced bone loss can be both prevented and rescued by a CB(2) specific agonist. Hence, synthetic CB(2) ligands, which are stable and orally available, provide a basis for developing novel anti-osteoporotic therapies, free of psychotropic effects. The CNR2 gene (encoding CB(2)) in women is associated with low bone mineral density, offering an assay for identifying females at risk of developing osteoporosis. PMID- 18426504 TI - Multiple roles for the endocannabinoid system during the earliest stages of life: pre- and postnatal development. AB - The endocannabinoid system, including its receptors (CB(1) and CB(2)), endogenous ligands ('endocannabinoids'), synthesising and degrading enzymes, as well as transporter molecules, has been detected from the earliest stages of embryonic development and throughout pre- and postnatal development. In addition, the endocannabinoids, notably 2-arachidonyl glycerol, are also present in maternal milk. During three distinct developmental stages (i.e. embryonic implantation, prenatal brain development and postnatal suckling), the endocannabinoid system appears to play an essential role for development and survival. Thus, during early pregnancy, successful embryonic passage through the oviduct and implantation into the uterus both require critical enzymatic control of optimal anandamide levels at the appropriate times and sites. During foetal life, the cannabinoid CB(1) receptor plays a major role in brain development, regulating neural progenitor differentiation into neurones and glia and guiding axonal migration and synaptogenesis. Postnatally, CB(1) receptor blockade interferes with the initiation of milk suckling in mouse pups, by inducing oral motor weakness, which exposes a critical role for CB(1) receptors in the initiation of milk suckling by neonates, possibly by interfering with innervation of the tongue muscles. Manipulating the endocannabinoid system by pre- and/or postnatal administration of cannabinoids or maternal marijuana consumption, has significant, yet subtle effects on the offspring. Thus, alterations in the dopamine, GABA and endocannabinoid systems have been reported while enhanced drug seeking behaviour and impaired executive (prefrontal cortical) function have also been observed. The relatively mild nature of the disruptive effects of prenatal cannabinoids may be understood in the framework of the intricate timing requirements and frequently biphasic effects of the (endo)cannabinoids. In conclusion, the endocannabinoid system plays several key roles in pre- and postnatal development. Future studies should further clarify the mechanisms involved and provide a better understanding of the adverse effects of prenatal exposure, in order to design strategies for the treatment of conditions such as infertility, mental retardation and failure-to-thrive. PMID- 18426505 TI - Interplay between endocannabinoids, steroids and cytokines in the control of human reproduction. AB - The use of marijuana, which today is the most used recreational drug, has been demonstrated to affect adversely reproduction. Marijuana smokers, both men and women, show impaired fertility, owing to defective signalling pathways, aberrant hormonal regulation, or wrong timing during embryo implantation. Anandamide (N arachidonoylethanolamine, AEA) and 2-arachidonoylglycerol (2-AG) mimic Delta(9) tetrahydrocannabinol (THC), the psychoactive principle of Cannabis sativa, by binding to both the brain-type (CB(1)) and the spleen-type (CB(2)) cannabinoid receptors. These 'endocannabinoids' exert several actions either in the central nervous system or in peripheral tissues, and are metabolised by specific enzymes that synthesise or hydrolyse them. In this review, we shall describe the elements that constitute the endocannabinoid system (ECS), in order to put in a better perspective the role of this system in the control of human fertility, both in females and males. In addition, we shall discuss the interplay between ECS, sex hormones and cytokines, which generates an endocannabinoid-hormone-cytokine array critically involved in the control of human reproduction. PMID- 18426506 TI - The cannabinoid system and male reproductive functions. AB - Cannabinoids, the main active components of marijuana, have been shown to exert different adverse effects on male reproduction both in vertebrates and invertebrates. In vivo, cannabinoids exert negative effects on hypothalamic hypophyseal reproductive hormone secretion and testicular endocrine and exocrine functions. Furthermore, a large amount of experimental data obtained in vitro have clearly shown that cannabinoids negatively influence important sperm functions, including motility and acrosome reaction, two fundamental processes necessary for oocyte fertilisation. These inhibitory effects are mediated by the direct action of cannabinoids on sperm through the activation of the cannabinoid receptor subtype CNR1 that has been shown to be expressed in mature sperm. In the present paper, we briefly review the effects of cannabinoids and endocannabinoids, a particular group of endogenously produced cannabinoids, on male reproductive function. PMID- 18426507 TI - Endocannabinoids and nutrition. AB - The endocannabinoids anandamide (AEA) and 2-arachidonoylglycerol (2-AG) are bioactive lipids derived from the n-6 family of polyunsaturated fatty acids that are essential fatty acids. Symptoms of essential fatty acid deficiency in rats - growth retardation, scaly skin, and increased transepidermal water loss - can mainly be attributed to lack of linoleic acid as a structural element of the epidermis. Arachidonic acid, however, also serve essential functions, particularly in cellular signalling via its precursor role for numerous oxygenated derivatives such as prostaglandins, leukotrienes, hepoxilins and other eicosanoids. Furthermore, arachidonic acid is also a structural part of endocannabinoids that have signalling functions in relation to modulation of neurotransmitter release, which might involve physiological and pathophysiological phenomena such as regulation of appetite, energy metabolism, pain perception, memory and learning. Furthermore, along with AEA formation other acylethanolamides are always formed - e.g., oleoylethanolamide (OEA), that can inhibit food intake, and palmitoylethanolamide, that is anti-inflammatory - possibly through activation of peroxisome proliferator activated receptor alpha (PPAR alpha) and/or GPR119. As all these unsaturated fatty acids are ingested daily in smaller or larger amounts, one can ask whether different dietary fats can affect the levels of these fatty acids in the tissues and thereby the quantitative formation of these bioactive signalling molecules. Generally, in vivo arachidonic-acid-derived eicosanoid production can be increased and decreased by prolonged feeding with pharmacological levels of arachidonic acid and long-chain (n-3) fatty acids (fish oil), respectively. Changes in levels of these two fatty acids within the traditional human diet hardly affects the eicosanoid production, however. Moreover, preliminary data suggest that dietary intake of arachidonic acid and fish oil also doesn't easily affect endocannabinoid formation; however, dietary fat in terms of saturated, polyunsaturated and monounsaturated seems to affect tissue levels of AEA, 2-AG and OEA. PMID- 18426508 TI - Endocannabinoids: some like it fat (and sweet too). AB - There is growing interest in the commercialisation of the CB(1) receptor antagonist Rimonabant in Europe for the treatment of obesity and the metabolic syndrome. Clinical trials have shown that CB(1) receptor blockers are able to reduce not only food intake but also abdominal adiposity and its metabolic sequelae. Accordingly, CB(1) receptors, and tissue concentrations of endocannabinoids sufficient to activate them, are present in all brain and peripheral organs involved in the control of energy balance, including the hypothalamus, nucleus accumbens, pancreas, adipose tissue, skeletal muscle and liver. At the central level, the endocannabinoid system seems to play a dual role in the regulation of food intake by hedonic and homeostatic energy regulation. At the peripheral level, the endocannabinoid system seems to behave as a system that reduces energy expenditure and directs energy balance towards energy storage into fat. The emerging role of the endocannabinoid system in energy balance at both central and peripheral levels will be discussed in this review. PMID- 18426509 TI - Dysregulation of the endocannabinoid system in obesity. AB - An activation of the endocannabinoid system (ECS) in obesity with increased concentrations of endocannabinoids in several tissues and in the circulation is described in this review. This increased availability of endocannabinoids might stimulate cannabinoid receptors in a pathophysiological manner. The successful use of the cannabinoid receptor CB(1) inverse agonists rimonabant and taranabant for weight loss and the treatment of obesity-associated metabolic disorders might well be through blocking this overstimulation of cannabinoid receptors. At present, no single mechanism has been identified that explains the increased bioavailability of endocannabinoids in obesity. Both increased synthesis and decreased degradation appear to operate in a species- and tissue-dependent manner, but many pieces of the puzzle still need to be collected. For example, most data show decreased fatty acid amide hydrolase (FAAH) expression and/or activity as a result of obesity or high-fat intake, but the endocannabinoid predominantly increased in tissues is 2-arachidonoylglycerol (2-AG), which is not degraded by FAAH in vivo. Furthermore, the influence of dietary fatty acids on the synthesis of endocannabinoids needs to be studied in much more detail. Although weight loss does not seem to influence activation of the endocannabinoid system (ECS) in human obesity, suggesting an underlying mechanisms independent of body weight, no such mechanism at the genetic level has yet been identified either. Thus, activation of the ECS is a hallmark of abdominal obesity, and explains the success of pharmacological CB(1) blockade, but serious attempts have to be made to clarify the underlying mechanisms of this activation. PMID- 18426510 TI - Central versus peripheral antagonism of cannabinoid CB1 receptor in obesity: effects of LH-21, a peripherally acting neutral cannabinoid receptor antagonist, in Zucker rats. AB - The endogenous cannabinoid system plays an important modulatory role in feeding behaviour and metabolism, acting at both central and peripheral levels. Chronic administration of cannabinoid CB(1) receptor antagonists has been found to be effective in experimental obesity. However, clinically available cannabinoid receptor antagonists are inverse agonists that can target CB(1) receptors located in both central circuits regulating appetite and motivation and in peripheral organs regulating metabolism and energy expenditure. This profile complicates understanding of cannabinoid CB(1) receptor blockade as a therapeutic strategy in obesity and metabolic disorders. This review aims to explore the relevance of both inverse agonism and peripheral cannabinoid receptor blockade on the beneficial actions of chronic cannabinoid receptor blockade, by comparing the actions of the reference antagonist/inverse agonist rimonabant and the newly designed drug LH-21. LH-21 is a triazol derivative and a neutral cannabinoid receptor antagonist; it has a poor penetration rate into the central nervous system. When given acutely it decreases food intake and enhances the anorectic actions of oleoylethanolamide, a feeding suppressant lipid that acts on peripheral sensory terminals in a similar way as rimonabant. Unlike rimonabant, chronic administration of LH-21 (3 mg/kg) reduces feeding but does not improve hypertriglyceridaemia or hypercholesterolaemia; nor does it reduce liver fat deposits in Zucker rats. These results suggest that the inverse agonism and/or the antagonism of central cannabinoid CB(1) receptors are necessary for the metabolic benefits of cannabinoid CB(1) receptor blockade, but not for the appetite reduction. PMID- 18426511 TI - Endocannabinoids, adipose tissue and lipid metabolism. AB - Endocannabinoids regulate energy balance by modulating hypothalamic circuits controlling food intake and energy expenditure. However, convincing evidence has accumulated indicating that the endocannabinoid system is present also in peripheral tissues, in particular in adipose tissue. Fat cells produce (and are targets of) endocannabinoids. Adipogenesis, lipogenesis and glucose uptake are stimulated by endocannabinoids through CB(1) receptors and these effects are blocked by the CB(1) receptor antagonist rimonabant, suggesting that the weight lowering effect of CB(1) receptor blockade is partly due to peripheral mechanisms. This review will focus on the role of endocannabinoids in adipose tissue metabolism, adipokine production and interactions between endocannabinoids and peroxisome proliferator activated receptors during adipogenesis. PMID- 18426512 TI - Cannabinoid type 1 receptor: another arrow in the adipocytes' bow. AB - The endocannabinoid system has recently emerged as an important modulator of several functions of adipose tissue, including cell proliferation, differentiation and secretion. Here, we will review the effects of cannabinoid type 1 (CB(1)) receptor activation/blockade in adipocytes by summarising the data in the literature since the discovery of the presence of this receptor in adipose tissue. We will also discuss our original data obtained in mouse 3T3-L1 adipocyte cells using WIN55 212, a CB(1)/CB(2) receptor agonist and SR141716 (rimonabant), a specific CB(1) receptor antagonist, respectively, in different experimental settings. PMID- 18426513 TI - CB1 receptor blockade and its impact on cardiometabolic risk factors: overview of the RIO programme with rimonabant. AB - Rimonabant, the first selective CB(1) receptor antagonist in clinical use, has been extensively investigated in the Rimonabant in Obesity (RIO) programme, comprising four 1-2 year placebo-controlled randomised clinical trials recruiting more than 6600 overweight/obese patients with or without co-morbidities. Rimonabant 20 mg daily consistently reduced body weight, waist circumference, triglycerides, blood pressure, insulin resistance and C-reactive protein levels, and increased HDL cholesterol concentrations in both non-diabetic and type-2 diabetic overweight/obese patients. Adiponectin levels were increased, an effect that correlated with HDL cholesterol augmentation, while small dense LDL cholesterol levels were decreased in patients receiving rimonabant 20 mg compared with those receiving placebo in RIO Lipids. Furthermore, in RIO Diabetes, a 0.7% reduction in glycated haemoglobin (HbA1c) levels was observed in metformin- or sulphonylurea-treated patients with type-2 diabetes, an effect recently confirmed in the 6-month SERENADE (Study Evaluating Rimonabant Efficacy in drug-NAive DiabEtic patients) trial in drug-naive diabetic patients. Almost half of metabolic changes occurred beyond weight loss, in agreement with direct peripheral effects. The positive effects observed after 1 year were maintained after 2 years. Rimonabant was generally well-tolerated, but with a slightly higher incidence of depressed mood disorders, anxiety, nausea and dizziness compared with placebo. In clinical practice, rimonabant has to be prescribed to the right patient, i.e. overweight/obese subjects with cardiometabolic risk factors and with no major depressive illness and/or ongoing antidepressive treatment, in order to both maximise efficacy and minimise safety issues. New trials are supposed to confirm the potential role of rimonabant in patients with abdominal adiposity, atherogenic dyslipidaemia and/or type-2 diabetes, i.e. at high cardiometabolic risk. PMID- 18426514 TI - The endocannabinoid system and the control of glucose homeostasis. AB - Blockade of the CB(1) receptor is one of the promising strategies for the treatment of obesity. The first selective CB(1) receptor antagonist, rimonabant, which has already successfully completed phase III clinical trials, led to sustained weight loss and a reduction in waist circumference. Patients treated with rimonabant also demonstrated statistically significant improvement in high density lipoprotein cholesterol levels, triglyceride levels and insulin resistance, as well as a reduced overall prevalence of metabolic syndrome. Currently, one of the most discussed aspects of endocannabinoid system function is to what extent the endocannabinoid system might affect metabolism independently of its control over body weight and food intake. Specifically, a food-intake- and body-weight-independent role in the regulation of glucose homeostasis and insulin sensitivity could have major impact on the potential of drug candidates targeting the endocannabinoid system for the prevention and treatment of metabolic syndrome. This review summarises the effects of the endocannabinoid system on glucose homeostasis and insulin sensitivity. PMID- 18426515 TI - Prenatal polychlorinated biphenyl exposures in eastern Slovakia modify effects of social factors on birthweight. AB - Polychlorinated biphenyls (PCB) were widely used for industrial purposes and consumer products, but because of their toxicity, production was banned by most industrialised countries in the late 1970s. In eastern Slovakia, they were produced until 1985. During 2002-04, a birth cohort of mothers (n = 1057) residing in two Slovak districts was enrolled at delivery, and their specimens and information were collected after birth. Congeners of PCBs were measured in maternal serum by high-resolution gas chromatography with electron capture detection. In this study, we used multiple linear regression to examine the effects of prenatal PCB exposure on birthweight adjusted for gestational age, controlling for inter-pregnancy interval, and maternal smoking, age, education, ethnicity, pre-pregnancy body mass index and height. The association between total maternal serum PCB levels and birthweight was not statistically significant. However, an interaction model indicated that maternal PCB concentrations were associated with lower birthweight in Romani boys. Based on the fitted regression model, the predicted birthweight of Romani boys at the 90th percentile of maternal PCBs (12.8 ng/mL) was 133 g lower than the predicted birthweight at the 10th percentile of maternal PCBs (1.6 ng/mL). This is a similar magnitude of effect to that observed for maternal smoking and birthweight. These results suggest that higher levels of PCBs in maternal blood sera may inhibit growth in boys, particularly in those already affected by social factors related to ethnicity. This study is consistent with previous findings that boys are more susceptible than girls to growth restriction induced by in utero organochlorine exposures, and further indicates that high PCBs may magnify the influence of social disadvantage in this vulnerable group of boys. PMID- 18426516 TI - Influences of study design and location on the relationship between particulate matter air pollution and birthweight. AB - A large number of studies have identified a relationship between particulate matter air pollution and birthweight. Although reported associations are small and varied, they have been identified in studies from places around the world. Exposure assignment, covariates and study inclusion criteria vary among studies. To examine the effect of these and other study characteristics on associations between particulate matter and birthweight, US birth records for singletons delivered at 40 weeks gestation in 2001-03 during the months of March, June, September and December were linked to quarterly estimates of pollution exposure, both particulate matter exposure and exposure to multiple pollutants, by county of residence and month of birth. Annual, 9-month and trimester-specific exposures were assigned. Among births linked to particulate matter exposure there was a small association between coarse particle exposure and birthweight (beta -13 g per 10 microg/m(3) increase [95% CI -18.3 g, -7.6 g]) after controlling for maternal factors; this association was attenuated slightly and remained statistically significant after further adjustment for contextual factors, year of birth, region, or urban-rural status. The associations were slightly weaker among births linked to multiple pollutant exposure than among births linked to just particulate matter exposure. The association varied markedly by region, ranging from a decrement of 43 g per 10 microg/m(3)[95% CI -58.6 g, -27.6 g] in the north-west to a null association in the south-west. Trimester findings were smaller, yet remained significant and varied regionally. The association between fine particle exposure and birthweight varied considerably, with an overall small positive association that became null after control for region. This study found that wide regional differences in association may contribute to the varied published findings. The association between coarse particle exposure and birthweight appeared robust, if small; fine particles had no overall association with birthweight. PMID- 18426519 TI - Biopsychosocial determinants of pregnancy length and fetal growth. AB - The causes and mechanisms related to preterm delivery and intrauterine growth restriction are poorly understood. Our objective was to assess the direct and indirect effects of psychosocial and biomedical factors on the duration of pregnancy and fetal growth. A self-administered questionnaire was distributed to pregnant women attending prenatal ultrasound clinics in nine hospitals in the Monteregie region in the province of Quebec, Canada, from November 1997 to May 1998. Prenatal questionnaires were linked with birth certificates. Theoretical models explaining pregnancy length and fetal growth were developed and tested, using path analysis. In order to reduce the number of variables from the questionnaire, a principal component analysis was performed, and the three most important new dimensions were retained as explanatory variables in the final models. Data were available for 1602 singleton pregnancies. The biophysical score, covering both maternal age and the pre-pregnancy body mass index, was the only variable statistically associated with pregnancy length. Smoking, obstetric history, maternal health and biophysical indices were direct predictors of fetal growth. Perceived stress, social support and self-esteem were not directly related to pregnancy outcomes, but were determinants of smoking and the above mentioned biomedical variables. More studies are needed to identify the mechanisms by which adverse psychosocial factors are translated into adverse biological effects. PMID- 18426518 TI - Placental characteristics and birthweight. AB - Standard gross placental measures capture dimensions relevant to specific placental functions. Our objective was to determine their accountability independent of placental weight for variance in birthweight, an important proxy for intrauterine 'adequacy' in fetal origins studies. The sample consisted of 24 152 singleton liveborn children of the Collaborative Perinatal Project delivered from 34 to 42 completed weeks gestation, with complete data for six placental measures (placental disc shape, umbilical cord length, distance from cord insertion to nearest margin, large diameter, small diameter, placental thickness) and placental weight. Associations between birthweight and placental measures were examined using multiple linear regression. Placental weight alone accounted for 36.6% of birthweight variation; the six other placental measures accounted for 28.1%. Combined, all placental measures accounted for 39.1% of birthweight variation. Seven maternal characteristics (age, height, weight, parity, socio economic status, cigarette use, and race) were investigated to determine whether their known associations with birthweight were mediated by placental markers. Analysis suggested that the impact of all maternal characteristics except smoking was consistent with mediation by placental characteristics. PMID- 18426520 TI - Maternal diet and the risk of hypospadias and cryptorchidism in the offspring. AB - Male genital tract birth defects have been associated in previous studies with several prenatal exposures to environmental and dietary risk factors. The purpose of this study was to explore the association between hypospadias and cryptorchidism, and the dietary habits of an agricultural population in Italy. A population-based case-control study was conducted in the Sicilian Province of Ragusa. Cases (n = 90) and controls (n = 202) included births for the period 1998 2002. Data on dietary habits of the mothers, as well as health-related social, occupational and environmental exposures prior to and during the index birth, were collected through interviews. Adjusted odds ratios (OR) were calculated by logistic regression after adjustment for confounding variables. Increased ORs were observed for mothers of children with hypospadias who, during pregnancy, frequently consumed fish (OR = 2.33 [95% confidence interval (CI) 1.03, 5.31]) and market-purchased fruit (OR = 5.10 [95% CI 1.31, 19.82]). For cryptorchidism, increased risk was observed in mothers consuming liver (OR = 5.21 [95% CI 1.26, 21.50]), and smoked products (OR = 2.46 [95% CI 1.15, 5.29]). For the two malformations pooled together, increased risk was associated with maternal consumption of liver (OR = 4.38 [95% CI 1.34, 14.26]) and with frequent consumption of wine (OR = 1.98 [95% CI 1.01, 3.86]). This study suggests that some maternal dietary factors may play a role in the development of congenital defects of the male reproductive tract. In particular, our data indicate that further research may be warranted on the endocrine-disrupting effects resulting from the bioaccumulation of contaminants (fish, liver), pesticides (marketed fruit, wine) and/or potentially toxic food components (smoked products, wine, liver). PMID- 18426521 TI - Measuring dietary sodium intake in infancy: a review of available methods. AB - It has been hypothesised that infancy may be a particularly sensitive period with respect to the effect of dietary sodium (salt) intake on later blood pressure. If this is the case, interventions to reduce dietary sodium intake in infancy could have major public health impact by reducing cardiovascular disease in the future. However, research in this area is hampered by difficulties of how to measure dietary sodium intake in infancy. Here we review the literature with an aim to describe different methods that have been used for research purposes, how these have been used and their relative strengths and limitations. We identified five studies that were relevant to our aim. In epidemiological and intervention studies sodium intake in infancy has been assessed via parents using diet diaries, which appear to be feasible and acceptable to parents. In these studies, sodium concentration in breast milk is assumed not to vary from one woman to the next, which may not be correct but has been poorly studied. The gold standard method - 24 h urinary excretion of sodium - was used in only one study and there is debate about the most appropriate way to collect urine for research purposes in infants. None of the studies directly compared different methods for determining infant dietary sodium intake. We conclude that research is required to determine the best methods of estimating dietary sodium intake in infants in different research contexts. PMID- 18426522 TI - Trends in stillbirths, early and late neonatal mortality in rural Bangladesh: the role of public health interventions. AB - Trends were examined in a cohort study of stillbirths and early and late neonatal deaths in Matlab, a rural area of Bangladesh between 1975 and 2002, using routinely collected demographic surveillance data. Main outcome measures were stillbirths per 1000 births, early neonatal deaths per 1000 livebirths, and late neonatal deaths per 1000 children surviving after 1 week. We performed a logistic regression examining trends over time and between two areas in the three outcome measures, controlling for the effects of parental education, religion, time, geography, parity, maternal age and birth spacing. There was a marked decline in stillbirths, early and late neonatal mortality over time in both areas, though the pace of decline was somewhat faster in the ICDDR,B (International Centre for Diarrhoeal Disease Research, Bangladesh) service area. Stillbirths declined by 24% overall in the ICDDR,B service area (crude OR comparing 1996-2002 with 1975 81: 0.76 [95% CI 0.68, 0.84]), compared with 15% in the Government service area (crude OR comparing 1996-2002 with 1975-81: 0.85 [0.76, 0.94]). The overall reduction in early and late neonatal mortality comparing the same periods was 39% and 73%, respectively, in the ICDDR,B area, compared with 30% and 63%, respectively, in the Government service area. Adjusting for socio-economic or demographic factors did not substantially alter the time or area differentials. The dramatic decline in neonatal mortality was, in large part, due to a fall in deaths from neonatal tetanus. The pace of decline was faster in the area receiving intense maternal and child health and family planning interventions, but stillbirths, early and late neonatal deaths also declined in the area not receiving such intense attention, suggesting that factors outside the formal health sector play an important role. PMID- 18426523 TI - Pre-conception counselling in primary care: prevalence of risk factors among couples contemplating pregnancy. AB - The outcome of pregnancy can be influenced by several risk factors. Women who are informed about these risks during pre-conception counselling (PCC) have an opportunity to take preventive measures in time. Several studies have shown that high-risk populations have a high prevalence of such risk factors. However, prevalence in the general population, which is assumed to be low risk, is largely unknown. We therefore provided a systematic programme of PCC for the general population and studied the prevalence of risk factors using the risk-assessment questionnaire which was part of the PCC. None of the couples reported no risk factors at all and only 2% of the couples reported risk factors for which written information was considered to be sufficient. Therefore, 98% of all couples reported one or more risk factors for which at least personal counselling by a general practitioner (GP) was indicated. Many of these factors were related to an unhealthy lifestyle. Women with a low level of education reported more risk factors than women with a high level of education. There is a great need for PCC as shown by the fact that almost all couples reported risk factors for which personal counselling was indicated. Pre-conception counselling may reduce the risk of adverse pregnancy outcome by enabling couples to avoid these risks. PCC can be provided by GPs, who have the necessary medical knowledge and background information to counsel couples who wish to have a baby. PMID- 18426524 TI - Vitamin supplement use among children with Down's syndrome and risk of leukaemia: a Children's Oncology Group (COG) study. AB - Vitamin supplements have been proposed for children with Down's syndrome (DS) with claims of improving cognitive abilities, or immune or thyroid function. Several studies have shown decreased levels of zinc in this population. Because children with DS have a 50-fold increased risk of developing acute leukaemia during the first 5 years of life, we explored the relation between child vitamin and herbal supplement use and the risk of leukaemia in a case-control study. During the period 1997-2002, we enrolled 158 children with DS aged 0-18 years that were diagnosed with acute lymphoblastic leukaemia (ALL) (n = 97) or acute myeloid leukaemia (AML) (n = 61) at participating Children's Oncology Group institutions. We enrolled 173 DS children without leukaemia (controls), selected from the cases' primary care clinic and frequency-matched on age. Data were collected via telephone interviews with mothers of the index child regarding use of multivitamins, zinc, vitamin C, iron and herbal supplements, including age at first use, frequency and duration. Among controls, 57% reported regular multivitamin use (>/=3 times/week for >/=3 months) compared with 48% of ALL cases and 61% of AML cases. We found no evidence of an association between children's regular multivitamin use and ALL or AML (adjusted odds ratios [OR] = 0.94 [95% CI 0.52, 1.70] and 1.90 [0.73, 4.91] respectively). There was a suggestion of an increased risk for AML associated with regular multivitamin use during the first year of life or for an extended duration (ORs = 2.38 [0.94, 5.76] and 2.59 [1.02, 6.59] respectively). Despite being the largest study of DS-leukaemia, our sample size was small, resulting in imprecise effect estimates. Future research should include larger sample sizes as well as a full assessment of diet including vitamin supplementation to adequately examine the relation between nutritional status and childhood leukaemia. PMID- 18426525 TI - Comparison of urinary and plasma cotinine levels during the three trimesters of pregnancy. AB - Assays of metabolised cotinine are considered to be an accurate measure of exposure to cigarette smoke among pregnant women. We investigated the association and differences between the cotinine levels in maternal urine and blood, and the umbilical cord blood of three tobacco exposure groups at different stages of pregnancy. A prospective study was conducted among 398 pregnant women undergoing prenatal care in different trimesters at two medical centres and one regional hospital in central Taiwan. All 398 subjects (including 25 smokers, 191 passive smokers and 182 non-smokers) remained in the study up to the time of delivery; 384 of them delivered singleton live births. Cotinine levels were assayed in the maternal plasma and urine of the mothers at each trimester and in the cord blood of the newborns. All specimens were measured using a sensitive high-performance liquid chromatography. Cotinine concentrations in plasma and urine showed a significant dose-dependent difference among the three groups (non-smoker, passive and active smoker) and a trend that increased with gestation among the pregnant women. Significant correlations between cotinine concentrations in plasma and urine among the pregnant women in each trimester were found. In addition, the level of cotinine in umbilical cord blood was significantly correlated with that in maternal blood at term (r = 0.89, P < 0.001). A pattern of elevated cotinine concentrations in the plasma and urine of pregnant women from the beginning to the end of pregnancy was found, and this correlated significantly with the cotinine levels in the umbilical cord blood. PMID- 18426526 TI - Correspondence between Helicobacter pylori antibodies and urea breath test results in a US-Mexico birth cohort. AB - The uncertain accuracy of methods for detecting Helicobacter pylori infection in young children complicates research on this infection in early life. The aim of the present report was to describe the correspondence between positive serology and positive urea breath test (UBT) in children followed from age 0 to 24 months in the Pasitos Cohort Study, conducted along the US-Mexico border at El Paso and Juarez. Children were recruited before birth during 1998-2000 and examined at target ages of 6, 12, 18 and 24 months. H. pylori infection was detected using an enzyme immunoassay for serum immunoglobulin G antibodies and the (13)C-urea breath test corrected for age-dependent variation in CO(2) production. Of 472 children, 125 had one or more positive UBT results and 46 had one or more positive serology results. The prevalence of H. pylori infection at target ages of 6, 12, 18 and 24 months was 7%, 14%, 16% and 19%, respectively, by UBT and 8%, 2%, 3% and 3%, respectively, by serology. Few (<1%) of those tested on both tests were positive on both at any age. Among UBT-positive children, 6% were concurrently seropositive and 6% became seropositive later. Because UBT positivity cut points were selected to minimise false positives, these results suggest that H. pylori infection occurred frequently in this cohort, but rarely produced detectable antibodies. For clinical or epidemiological investigations, serology should not be used as the sole method for detecting H. pylori infection in children aged 2 years or less. PMID- 18426528 TI - Plant form and function. PMID- 18426529 TI - Stealth tactics of galling parasites and their potential indirect effects. PMID- 18426530 TI - The genetic diversity of intraterrestrial aliens. PMID- 18426531 TI - Whither plant evo-devo? PMID- 18426536 TI - Destiny matters: distal developmental influences on adult alcohol use and abuse. AB - A foundational assumption in the fields of addiction and developmental psychopathology is that child and adolescent experiences set the stage for adult functioning and adjustment. However, the empirical literature documenting life span linkages with adult alcohol (and other drug) use and abuse is sparse. This gap is due to a slow adoption of life-span developmental conceptualizations and the lack of long-term prospective longitudinal studies. This supplemental issue provides evidence for such linkages from six long-term longitudinal studies, which together follow individuals from birth through to the late 40s. The data sets include national and regional samples from Britain, Finland and the United States. In this introductory paper, we consider conceptual issues concerning linkages across the life-span culminating in adult alcohol use and disorders, and provide a summary of the purposes and common themes. PMID- 18426537 TI - Childhood and adolescent predictors of alcohol use and problems in adolescence and adulthood in the National Child Development Study. AB - AIMS: To identify childhood and adolescent predictors of alcohol use and harmful drinking in adolescence and adulthood. DESIGN: Longitudinal data from childhood to mid-life from the National Child Development Study (NCDS) were used, including predictors collected at ages 7, 11, 16 years and alcohol outcomes collected at ages 16, 23, 33 and 42 years. SETTING: The NCDS is an ongoing longitudinal study of a cohort of 1 week's births in Britain in 1958. PARTICIPANTS: Childhood and adolescent predictors and alcohol use data from at least one adolescent or adult wave were available from 7883 females and 8126 males. MEASUREMENTS: Social background, family, academic and behavioural predictors measured at ages 7, 11 and 16 years were entered into hierarchical multiple and logistic regressions to predict quantity of alcohol use at ages 16, 23, and 33 years and harmful drinking [i.e. Cut-down, Annoyed, Guilt, Eye-opener (CAGE) questionnaire score] by age 42 years. FINDINGS: Previous drinking was controlled in final models to predict change. Drinking was heavier among those with greater childhood and adolescent social advantage (especially females), less harmonious family relationships, more social maladjustment, greater academic performance, less internalizing problems, more truancy and earlier school-leaving plans. CONCLUSIONS: Alcohol use and problems in adulthood can be predicted by indicators of social background, adjustment and behaviour in childhood and adolescence. Results demonstrate that the early roots of adolescent and adult alcohol use behaviours begin in childhood. PMID- 18426539 TI - Childhood and adolescent predictors of early and middle adulthood alcohol use and problem drinking: the Columbia County Longitudinal Study. AB - AIMS: To examine the role of individually and contextually based factors measured during childhood and adolescence in predicting alcohol use and abuse measured during early and middle adulthood. DESIGN: Initial sample of 856 individuals first interviewed at age 8 with follow-up interviews at 19 (n = 427), 30 (n = 409), and 48 (n = 523). PARTICIPANTS: Individuals enrolled in 3rd-grade classes in Columbia County, NY, in 1960 (49% female; > 90% Caucasian; primarily working class families), who were re-sampled in 1970 (51% female), 1981 (52% female) and 2000 (49% female). MEASUREMENTS: Parent reports of negative family interaction and socio-economic status when the child was 8 years old; IQ test at age 8; peer nominations of aggression, popularity and behavioral inhibition at ages 8 and 19; self-report of depression and educational attainment at age 19; self-report of alcohol use and problem drinking at ages 30 and 48. FINDINGS: Path models showed that the effects of childhood individual variables (e.g. aggression, popularity, behavioral inhibition) on adulthood alcohol use and abuse generally were mediated by the same behavioral variables in adolescence. Specifically, both for males and for females, lower levels of behavioral inhibition and higher levels of aggression predicted adulthood alcohol variables. Childhood contextual variables (family socio-economic status and negative family interaction) were relatively weak predictors of adulthood alcohol use and abuse. CONCLUSIONS: Alcohol use and abuse in adulthood, when considered in a long-term developmental-contextual framework, appear to be consistent with a general deviance model of problem behavior whereby individually based factors from childhood and late adolescence predict long-term indices of adulthood alcohol use and abuse. PMID- 18426538 TI - Childhood and adolescent predictors of heavy drinking and alcohol use disorders in early adulthood: a longitudinal developmental analysis. AB - AIMS: To identify childhood and adolescent factors differentiating heavy alcohol users in early adulthood from more moderate users or abstainers. DESIGN: Low income participants followed from birth to age 28 years. PARTICIPANTS: A total of 178 adults (95 males) who were first-born children of low-income mothers recruited in Minneapolis, Minnesota, during their third trimester of pregnancy. MEASUREMENTS: Maternal hostility (24/42 months), externalizing and internalizing behavior problems (9 years), peer acceptance and academic achievement (12 years), maternal alcohol use and participants' drinking behavior (16 years), quantity of alcohol use per occasion (19, 23 and 26 years), alcohol use disorders (28 years). FINDINGS: For men: (i) higher amounts of alcohol consumption at age 16 increased the odds of being a heavy drinker compared to an abstainer (age 19) and a moderate drinker (ages 23 and 26); (ii) lower achievement scores at age 12 and having a mother who drank more when the participant was age 16 increased the odds of being a heavy drinker compared to moderate drinker (age 26). Higher levels of externalizing behavior problems at age 9 and drinking more when the participants were age 16 increased the odds that men would have a current alcohol use disorder at age 28. For women: (i) drinking more at age 16 increased the odds of being a heavy drinker compared to being either an abstainer or a moderate drinker (age 26); (ii) having higher levels of achievement at age 12 increased the odds of being a heavy drinker compared to an abstainer at age 23. Adolescent alcohol use mediated the relation between externalizing behavior at age 9 and alcohol use at age 26 for women. CONCLUSIONS: Problem drinking may be the result of a long-term developmental process wherein childhood externalizing behavior problems sets a pathway leading to heavy drinking during and after adolescence. PMID- 18426540 TI - A developmental approach to alcohol drinking behaviour in adulthood: a follow-up study from age 8 to age 42. AB - AIMS: To study the links of family background, child and adolescent social behaviour, and (mal)adaptation with heavy drinking by age 20 and with the frequency of drinking, binge drinking, Cut-down, Annoyed, Guilt, Eye-opener (CAGE) questionnaire scores and problems due to drinking at ages 27 and 42 years. DESIGN: In the Finnish Jyvaskyla Longitudinal Study of Personality and Social Development, data have been collected by interviews, inventories and questionnaires. Behavioural data were gathered at ages 8 and 14; data on alcohol consumption were gathered at ages 14, 20, 27, 36 and 42. PARTICIPANTS: A total of 184 males and 163 females; 94% of the original sample of the 8-year-olds. FINDINGS: Family adversities, externalizing problem behaviours, low school success, truancy and substance use in adolescence were associated in early middle age with problems due to drinking in both genders, and to binge drinking and CAGE scores in females. The antecedents varied, however, across the indicators of drinking and gender. The frequency of drinking was least predictable by the studied antecedents. Childhood and adolescent antecedents and drinking up to age 20 explained 43% of males' and 31% of females' problem drinking at age 42; 31% and 19%, respectively, at age 27. CONCLUSIONS: The early warning signs of drinking problems should be taken seriously in the preventive work for alcohol abuse. Problem drinking in early middle age is preceded by maladjustment to school, early age of onset of drinking and heavy drinking in adolescence even more significantly than problem drinking in early adulthood. PMID- 18426541 TI - Adolescent pathways to adulthood drinking: sport activity involvement is not necessarily risky or protective. AB - AIMS: Use pattern-centered methods to examine how adolescents' alcohol use and sports activities are related both to childhood sport and problem behavior and to heavy drinking in early adulthood. DESIGN: The data used in this study come from four waves of the Michigan Study of Adolescent Life Transitions (MSALT) that began in 1983, when participants were approximately age 12, and continued into early adulthood, when participants were approximately age 28. PARTICIPANTS: Sixty per cent of the approximately 1000 MSALT youth living in south-eastern Michigan were females and 97% were European American. Approximately 28% of one or both parents held at least a college degree, and 45% held a high school diploma or lower. FINDINGS: Pattern-centered analyses revealed that the relation between adolescent sport activity and age 28 heavy alcohol use obtained primarily for sport participants who were also using more than the average amount of alcohol and other drugs at age 18. Similarly, children who were characterized by relatively high levels of sport participation, aggression and other problem behavior at age 12 were more likely than expected by chance to become sport participants who used more than the average amount of alcohol and other drugs at age 18. CONCLUSIONS: The results indicate that childhood problem behavior and adolescent sport participation can, but do not necessarily, presage heavy drinking in adulthood and that pattern-centered analytical techniques are useful for revealing such theoretically generated predictions. PMID- 18426542 TI - Adolescent risk factors for adult alcohol use and abuse: stability and change of predictive value across early and middle adulthood. AB - AIMS: To examine age-18 risk factors for alcohol use and heavy drinking during early (ages 22 and 26) and middle (age 35) adulthood, and for symptoms of alcohol use disorders (AUDs) in middle adulthood. DESIGN: Nationally representative samples of US adolescents in their senior year of secondary school (age 18) were followed into middle adulthood. Structural equation models estimated the associations between age-18 characteristics and current drinking and heavy drinking at ages 22, 26 and 35 and symptoms of AUDs at age 35. PARTICIPANTS: The sample consisted of 21,137 respondents from 11 senior year cohorts (1976-86) from the Monitoring the Future study. FINDINGS: Many predictor variables had stable associations with alcohol use over time, although their ability to explain variance in alcohol use declined with increasing time lags. Being white predicted alcohol use, but not symptoms of AUDs. Parental drinking, risk taking and use of cigarettes and marijuana predicted heavy drinking to age 35. Planning to attend college predicted more heavy drinking at age 22 and less frequent heavy drinking by mid-life. High school theft and property damage predicted later AUD symptoms. Most associations were invariant across gender, with variations typically taking the form of stronger associations between predictors and alcohol use for men. Invariance in findings across cohorts indicates that results reflect general developmental trends rather than specific historically bounded ones. CONCLUSIONS: Many adolescent individual and contextual characteristics remain important predictors of adult alcohol use and abuse, and their predictive impact varies as a function of age and type of alcohol outcome. These associations are largely equivalent across gender and cohort, thus reflecting robust developmental linkages. PMID- 18426544 TI - The Second International Symposium on Languages in Biology and Medicine (LBM) 2007. PMID- 18426543 TI - Anticipating problem alcohol use developmentally from childhood into middle adulthood: what have we learned? AB - This commentary reviews and comments on six major longitudinal studies from the United States, Great Britain and Finland, that test predictive models of drinking and problem drinking behavior across a developmental span of one to two generations. The large Ns, in two instances involving population samples, and the broad and study-overlapping variable domains make this collection of studies unique and of special interest vis-a-vis the issue of cross-study replicability of findings. Significant cross-study commonalities are noted, involving the strong cross-study replicability of an undercontrol/externalizing domain as both a childhood and adolescent predictor of problem drinking outcomes in early to middle adulthood, the relative autostability of heavy and problem use of alcohol over intervals of time as long as a generation, the utility of early drinking behavior as an index for later drinking outcomes, the relative parallelism (with some exceptions) of male and female findings, albeit with greater predictability of male over female drinking outcomes and the relatively tighter relational networks of drinking and other behavioral characteristics for males. This impressive group of quasi-replications also points the field to address several next-step questions, including: (i) the need to parse the undercontrol/externalizing domain to identify those subcomponential process characteristics that are causal to heavy and problem drinking outcomes; (ii) the need to develop models that will handle more effectively the uneven relationships of negative activity to drinking outcomes, in some instances operating protectively, in other instances operating as risk factors; (iii) the need for more carefully articulated, theoretically driven process models that will specify the ordering, developmental saliency and mediational properties of risk and protective factors as they come on line; and (iv) the need for more developmental testing of trait/context interaction models of problem drinking development. PMID- 18426545 TI - Automatic construction of rule-based ICD-9-CM coding systems. AB - BACKGROUND: In this paper we focus on the problem of automatically constructing ICD-9-CM coding systems for radiology reports. ICD-9-CM codes are used for billing purposes by health institutes and are assigned to clinical records manually following clinical treatment. Since this labeling task requires expert knowledge in the field of medicine, the process itself is costly and is prone to errors as human annotators have to consider thousands of possible codes when assigning the right ICD-9-CM labels to a document. In this study we use the datasets made available for training and testing automated ICD-9-CM coding systems by the organisers of an International Challenge on Classifying Clinical Free Text Using Natural Language Processing in spring 2007. The challenge itself was dominated by entirely or partly rule-based systems that solve the coding task using a set of hand crafted expert rules. Since the feasibility of the construction of such systems for thousands of ICD codes is indeed questionable, we decided to examine the problem of automatically constructing similar rule sets that turned out to achieve a remarkable accuracy in the shared task challenge. RESULTS: Our results are very promising in the sense that we managed to achieve comparable results with purely hand-crafted ICD-9-CM classifiers. Our best model got a 90.26% F measure on the training dataset and an 88.93% F measure on the challenge test dataset, using the micro-averaged F beta=1 measure, the official evaluation metric of the International Challenge on Classifying Clinical Free Text Using Natural Language Processing. This result would have placed second in the challenge, with a hand-crafted system achieving slightly better results. CONCLUSIONS: Our results demonstrate that hand-crafted systems - which proved to be successful in ICD-9-CM coding - can be reproduced by replacing several laborious steps in their construction with machine learning models. These hybrid systems preserve the favourable aspects of rule-based classifiers like good performance, and their development can be achieved rapidly and requires less human effort. Hence the construction of such hybrid systems can be feasible for a set of labels one magnitude bigger, and with more labeled data. PMID- 18426546 TI - Identification of transcription factor contexts in literature using machine learning approaches. AB - BACKGROUND: Availability of information about transcription factors (TFs) is crucial for genome biology, as TFs play a central role in the regulation of gene expression. While manual literature curation is expensive and labour intensive, the development of semi-automated text mining support is hindered by unavailability of training data. There have been no studies on how existing data sources (e.g. TF-related data from the MeSH thesaurus and GO ontology) or potentially noisy example data (e.g. protein-protein interaction, PPI) could be used to provide training data for identification of TF-contexts in literature. RESULTS: In this paper we describe a text-classification system designed to automatically recognise contexts related to transcription factors in literature. A learning model is based on a set of biological features (e.g. protein and gene names, interaction words, other biological terms) that are deemed relevant for the task. We have exploited background knowledge from existing biological resources (MeSH and GO) to engineer such features. Weak and noisy training datasets have been collected from descriptions of TF-related concepts in MeSH and GO, PPI data and data representing non-protein-function descriptions. Three machine-learning methods are investigated, along with a vote-based merging of individual approaches and/or different training datasets. The system achieved highly encouraging results, with most classifiers achieving an F-measure above 90%. CONCLUSIONS: The experimental results have shown that the proposed model can be used for identification of TF-related contexts (i.e. sentences) with high accuracy, with a significantly reduced set of features when compared to traditional bag-of-words approach. The results of considering existing PPI data suggest that there is not as high similarity between TF and PPI contexts as we have expected. We have also shown that existing knowledge sources are useful both for feature engineering and for obtaining noisy positive training data. PMID- 18426547 TI - Normalizing biomedical terms by minimizing ambiguity and variability. AB - BACKGROUND: One of the difficulties in mapping biomedical named entities, e.g. genes, proteins, chemicals and diseases, to their concept identifiers stems from the potential variability of the terms. Soft string matching is a possible solution to the problem, but its inherent heavy computational cost discourages its use when the dictionaries are large or when real time processing is required. A less computationally demanding approach is to normalize the terms by using heuristic rules, which enables us to look up a dictionary in a constant time regardless of its size. The development of good heuristic rules, however, requires extensive knowledge of the terminology in question and thus is the bottleneck of the normalization approach. RESULTS: We present a novel framework for discovering a list of normalization rules from a dictionary in a fully automated manner. The rules are discovered in such a way that they minimize the ambiguity and variability of the terms in the dictionary. We evaluated our algorithm using two large dictionaries: a human gene/protein name dictionary built from BioThesaurus and a disease name dictionary built from UMLS. CONCLUSIONS: The experimental results showed that automatically discovered rules can perform comparably to carefully crafted heuristic rules in term mapping tasks, and the computational overhead of rule application is small enough that a very fast implementation is possible. This work will help improve the performance of term-concept mapping tasks in biomedical information extraction especially when good normalization heuristics for the target terminology are not fully known. PMID- 18426548 TI - Assessment of disease named entity recognition on a corpus of annotated sentences. AB - BACKGROUND: In recent years, the recognition of semantic types from the biomedical scientific literature has been focused on named entities like protein and gene names (PGNs) and gene ontology terms (GO terms). Other semantic types like diseases have not received the same level of attention. Different solutions have been proposed to identify disease named entities in the scientific literature. While matching the terminology with language patterns suffers from low recall (e.g., Whatizit) other solutions make use of morpho-syntactic features to better cover the full scope of terminological variability (e.g., MetaMap). Currently, MetaMap that is provided from the National Library of Medicine (NLM) is the state of the art solution for the annotation of concepts from UMLS (Unified Medical Language System) in the literature. Nonetheless, its performance has not yet been assessed on an annotated corpus. In addition, little effort has been invested so far to generate an annotated dataset that links disease entities in text to disease entries in a database, thesaurus or ontology and that could serve as a gold standard to benchmark text mining solutions. RESULTS: As part of our research work, we have taken a corpus that has been delivered in the past for the identification of associations of genes to diseases based on the UMLS Metathesaurus and we have reprocessed and re-annotated the corpus. We have gathered annotations for disease entities from two curators, analyzed their disagreement (0.51 in the kappa-statistic) and composed a single annotated corpus for public use. Thereafter, three solutions for disease named entity recognition including MetaMap have been applied to the corpus to automatically annotate it with UMLS Metathesaurus concepts. The resulting annotations have been benchmarked to compare their performance. CONCLUSIONS: The annotated corpus is publicly available at ftp://ftp.ebi.ac.uk/pub/software/textmining/corpora/diseases and can serve as a benchmark to other systems. In addition, we found that dictionary look up already provides competitive results indicating that the use of disease terminology is highly standardized throughout the terminologies and the literature. MetaMap generates precise results at the expense of insufficient recall while our statistical method obtains better recall at a lower precision rate. Even better results in terms of precision are achieved by combining at least two of the three methods leading, but this approach again lowers recall. Altogether, our analysis gives a better understanding of the complexity of disease annotations in the literature. MetaMap and the dictionary based approach are available through the Whatizit web service infrastructure (Rebholz-Schuhmann D, Arregui M, Gaudan S, Kirsch H, Jimeno A: Text processing through Web services: Calling Whatizit. Bioinformatics 2008, 24:296-298). PMID- 18426549 TI - Exploiting and integrating rich features for biological literature classification. AB - BACKGROUND: Efficient features play an important role in automated text classification, which definitely facilitates the access of large-scale data. In the bioscience field, biological structures and terminologies are described by a large number of features; domain dependent features would significantly improve the classification performance. How to effectively select and integrate different types of features to improve the biological literature classification performance is the major issue studied in this paper. RESULTS: To efficiently classify the biological literatures, we propose a novel feature value schema TF*ML, features covering from lower level domain independent "string feature" to higher level domain dependent "semantic template feature", and proper integrations among the features. Compared to our previous approaches, the performance is improved in terms of AUC and F-Score by 11.5% and 8.8% respectively, and outperforms the best performance achieved in BioCreAtIvE 2006. CONCLUSIONS: Different types of features possess different discriminative capabilities in literature classification; proper integration of domain independent and dependent features would significantly improve the performance and overcome the over-fitting on data distribution. PMID- 18426550 TI - New challenges for text mining: mapping between text and manually curated pathways. AB - BACKGROUND: Associating literature with pathways poses new challenges to the Text Mining (TM) community. There are three main challenges to this task: (1) the identification of the mapping position of a specific entity or reaction in a given pathway, (2) the recognition of the causal relationships among multiple reactions, and (3) the formulation and implementation of required inferences based on biological domain knowledge. RESULTS: To address these challenges, we constructed new resources to link the text with a model pathway; they are: the GENIA pathway corpus with event annotation and NF-kB pathway. Through their detailed analysis, we address the untapped resource, 'bio-inference,' as well as the differences between text and pathway representation. Here, we show the precise comparisons of their representations and the nine classes of 'bio inference' schemes observed in the pathway corpus. CONCLUSIONS: We believe that the creation of such rich resources and their detailed analysis is the significant first step for accelerating the research of the automatic construction of pathway from text. PMID- 18426552 TI - Monitoring the evolutionary aspect of the Gene Ontology to enhance predictability and usability. AB - BACKGROUND: Much effort is currently made to develop the Gene Ontology (GO). Due to the dynamic nature of information it addresses, GO undergoes constant updates whose results are released at regular intervals as separate versions. Although there are a large number of computational tools to aid the development of GO, they are operating on a particular version of GO, making it difficult for GO curators to anticipate the full impact of particular changes along the time axis on a larger scale. We present a method for tapping into such an evolutionary aspect of GO, by making it possible to keep track of important temporal changes to any of the terms and relations of GO and by consequently making it possible to recognize associated trends. RESULTS: We have developed visualization methods for viewing the changes between two different versions of GO by constructing a colour coded layered graph. The graph shows both versions of GO with highlights to those GO terms that are added, removed and modified between the two versions. Focusing on a specific GO term or terms of interest over a period, we demonstrate the utility of our system that can be used to make useful hypotheses about the cause of the evolution and to provide new insights into more complex changes. CONCLUSIONS: GO undergoes fast evolutionary changes. A snapshot of GO, as presented by each version of GO alone, overlooks such evolutionary aspects, and consequently limits the utilities of GO. The method that highlights the differences of consecutive versions or two different versions of an evolving ontology with colour-coding enhances the utility of GO for users as well as for developers. To the best of our knowledge, this is the first proposal to visualize the evolutionary aspect of GO. PMID- 18426551 TI - Comparative analysis of five protein-protein interaction corpora. AB - BACKGROUND: Growing interest in the application of natural language processing methods to biomedical text has led to an increasing number of corpora and methods targeting protein-protein interaction (PPI) extraction. However, there is no general consensus regarding PPI annotation and consequently resources are largely incompatible and methods are difficult to evaluate. RESULTS: We present the first comparative evaluation of the diverse PPI corpora, performing quantitative evaluation using two separate information extraction methods as well as detailed statistical and qualitative analyses of their properties. For the evaluation, we unify the corpus PPI annotations to a shared level of information, consisting of undirected, untyped binary interactions of non-static types with no identification of the words specifying the interaction, no negations, and no interaction certainty. We find that the F-score performance of a state-of-the-art PPI extraction method varies on average 19 percentage units and in some cases over 30 percentage units between the different evaluated corpora. The differences stemming from the choice of corpus can thus be substantially larger than differences between the performance of PPI extraction methods, which suggests definite limits on the ability to compare methods evaluated on different resources. We analyse a number of potential sources for these differences and identify factors explaining approximately half of the variance. We further suggest ways in which the difficulty of the PPI extraction tasks codified by different corpora can be determined to advance comparability. Our analysis also identifies points of agreement and disagreement in PPI corpus annotation that are rarely explicitly stated by the authors of the corpora. CONCLUSIONS: Our comparative analysis uncovers key similarities and differences between the diverse PPI corpora, thus taking an important step towards standardization. In the course of this study we have created a major practical contribution in converting the corpora into a shared format. The conversion software is freely available at http://mars.cs.utu.fi/PPICorpora. PMID- 18426553 TI - Structuring an event ontology for disease outbreak detection. AB - BACKGROUND: This paper describes the design of an event ontology being developed for application in the machine understanding of infectious disease-related events reported in natural language text. This event ontology is designed to support timely detection of disease outbreaks and rapid judgment of their alerting status by 1) bridging a gap between layman's language used in disease outbreak reports and public health experts' deep knowledge, and 2) making multi-lingual information available. CONSTRUCTION AND CONTENT: This event ontology integrates a model of experts' knowledge for disease surveillance, and at the same time sets of linguistic expressions which denote disease-related events, and formal definitions of events. In this ontology, rather general event classes, which are suitable for application to language-oriented tasks such as recognition of event expressions, are placed on the upper-level, and more specific events of the experts' interest are in the lower level. Each class is related to other classes which represent participants of events, and linked with multi-lingual synonym sets and axioms. CONCLUSIONS: We consider that the design of the event ontology and the methodology introduced in this paper are applicable to other domains which require integration of natural language information and machine support for experts to assess them. The first version of the ontology, with about 40 concepts, will be available in March 2008. PMID- 18426554 TI - Gene Ontology density estimation and discourse analysis for automatic GeneRiF extraction. AB - BACKGROUND: This paper describes and evaluates a sentence selection engine that extracts a GeneRiF (Gene Reference into Functions) as defined in ENTREZ-Gene based on a MEDLINE record. Inputs for this task include both a gene and a pointer to a MEDLINE reference. In the suggested approach we merge two independent sentence extraction strategies. The first proposed strategy (LASt) uses argumentative features, inspired by discourse-analysis models. The second extraction scheme (GOEx) uses an automatic text categorizer to estimate the density of Gene Ontology categories in every sentence; thus providing a full ranking of all possible candidate GeneRiFs. A combination of the two approaches is proposed, which also aims at reducing the size of the selected segment by filtering out non-content bearing rhetorical phrases. RESULTS: Based on the TREC 2003 Genomics collection for GeneRiF identification, the LASt extraction strategy is already competitive (52.78%). When used in a combined approach, the extraction task clearly shows improvement, achieving a Dice score of over 57% (+10%). CONCLUSIONS: Argumentative representation levels and conceptual density estimation using Gene Ontology contents appear complementary for functional annotation in proteomics. PMID- 18426555 TI - Epigallocatechin-3-gallate (EGCG) inhibits the migratory behavior of tumor bronchial epithelial cells. AB - BACKGROUND: Many studies associated the main polyphenolic constituent of green tea, (-)-Epigallocatechin-3-gallate (EGCG), with inhibition of cancers, invasion and metastasis. To date, most of the studies have focused on the effect of EGCG on cell proliferation or death. Since cell migration is an important mechanism involved in tumor invasion, the aim of the present work was to target another approach of the therapeutic effect of EGCG, by investigating its effect on the cell migratory behavior. METHODS: The effect of EGCG (at concentrations lower than 10 microg/ml) on the migration speed of invasive cells was assessed by using 2D and 3D models of cell culture. We also studied the effects of EGCG on proteinases expression by RT-PCR analysis. By immunocytochemistry, we analyzed alterations of vimentin organization in presence of different concentrations of EGCG. RESULTS: We observed that EGCG had an inhibitory effect of cell migration in 2D and 3D cell culture models. EGCG also inhibited MMP-2 mRNA and protein expression and altered the intermediate filaments of vimentin. CONCLUSION: Taken together, our results demonstrate that EGCG is able to inhibit the migration of bronchial tumor cells and could therefore be an attractive candidate to treat tumor invasion and cell migration. PMID- 18426556 TI - The herpesvirus 8 encoded chemokines vCCL2 (vMIP-II) and vCCL3 (vMIP-III) target the human but not the murine lymphotactin receptor. AB - BACKGROUND: Large DNA-viruses such as herpesvirus and poxvirus encode proteins that target and exploit the chemokine system of their host. The Kaposi sarcoma- associated herpes virus (KSHV) encodes three chemokines. Two of these, vCCL2 and vCCL3, target the human lymphotactin receptor as an antagonist and a selective agonist, respectively. Therefore these virally endcoded chemokines have the potential to be used as tools in the study of lymphotactin receptor pathways in murine models. RESULTS: The activities of vCCL2, vCCL3, human lymphotactin (XCL1) and murine lymphotactin (mXCL1) were probed in parallel on the human and murine lymphotactin receptor (XCR1 and mXCR1) using a phosphatidyl-inositol assay. On the human XCR1, vCCL3, mXCL1 and XCL1 acted as agonists. In contrast, only mXCL1 was able to activate the murine lymphotactin receptor. Using the same assay, vCCL2 was able to block the response using any of the three agonists on the humane lymphotactin receptor with IC50s of 2-3 nM. However, vCCL2 was unable to block the response of mXCL1 through the murine lymphotactin receptor. CONCLUSION: This study shows that vCCL2 and vCCL3 cannot be used to investigate lymphotactin receptor pathways in murine models. These results also add vCCL2 and vCCL3 to a growing list of viral chemokines with known human chemokine receptor targets, which do not target the corresponding murine receptors. This fits with the observation that viral and endogenous ligands for the same human chemokine receptor tend to have relatively divergent amino-acid sequences, suggesting that these viruses have fine-tuned the design of their chemokines such that the action of the viral encoded chemokines cannot be expected to cross species barriers. PMID- 18426557 TI - A systematic survey in Arabidopsis thaliana of transcription factors that modulate circadian parameters. AB - BACKGROUND: Plant circadian systems regulate various biological processes in harmony with daily environmental changes. In Arabidopsis thaliana, the underlying clock mechanism is comprised of multiple integrated transcriptional feedbacks, which collectively lead to global patterns of rhythmic gene expression. The transcriptional networks are essential within the clock itself and in its output pathway. RESULTS: Here, to expand understanding of transcriptional networks within and associated to the clock, we performed both an in silico analysis of transcript rhythmicity of transcription factor genes, and a pilot assessment of functional phenomics on the MYB, bHLH, and bZIP families. In our in silico analysis, we defined which members of these families express a circadian waveform of transcript abundance. Up to 20% of these families were over-represented as clock-controlled genes. To detect members that contribute to proper oscillator function, we systematically measured rhythmic growth via an imaging system in hundreds of misexpression lines targeting members of the transcription-factor families. Three transcription factors were found that conferred aberrant circadian rhythms when misexpressed: MYB3R2, bHLH69, and bHLH92. CONCLUSION: Transcript abundance of many transcription factors in Arabidopsis oscillates in a circadian manner. Further, a developed pipeline assessed phenotypic contribution of a panel of transcriptional regulators in the circadian system. PMID- 18426558 TI - Oral and dental health care practices in pregnant women in Australia: a postnatal survey. AB - BACKGROUND: The aims of this study were to assess women's knowledge and experiences of dental health in pregnancy and to examine the self-care practices of pregnant women in relation to their oral health. METHODS: Women in the postnatal ward at the Women's and Children's Hospital, Adelaide, completed a questionnaire to assess their knowledge, attitudes and practices to periodontal health. Pregnancy outcomes were collected from their medical records. Results were analysed by chi-square tests, using SAS. RESULTS: Of the 445 women enrolled in the survey, 388 (87 per cent) completed the questionnaire. Most women demonstrated reasonable knowledge about dental health. There was a significant association between dental knowledge and practices with both education and socio economic status. Women with less education and lower socio-economic status were more likely to be at higher risk of poor periodontal health compared with women with greater levels of education and higher socioeconomic status. CONCLUSION: Most women were knowledgeable about oral and dental health. Lack of knowledge about oral and dental health was strongly linked to women with lower education achievements and lower socioeconomic backgrounds. Whether more intensive dental health education in pregnancy can lead to improved oral health and ultimately improved pregnancy outcomes requires further study. PMID- 18426559 TI - Diastolic time - frequency relation in the stress echo lab: filling timing and flow at different heart rates. AB - A cutaneous force-frequency relation recording system based on first heart sound amplitude vibrations has been recently validated. Second heart sound can be simultaneously recorded in order to quantify both systole and diastole duration. AIMS: 1- To assess the feasibility and extra-value of operator-independent, force sensor-based, diastolic time recording during stress. METHODS: We enrolled 161 patients referred for stress echocardiography (exercise 115, dipyridamole 40, pacing 6 patients).The sensor was fastened in the precordial region by a standard ECG electrode. The acceleration signal was converted into digital and recorded together with ECG signal. Both systolic and diastolic times were acquired continuously during stress and were displayed by plotting times vs. heart rate. Diastolic filling rate was calculated as echo-measured mitral filling volume/sensor-monitored diastolic time. RESULTS: Diastolic time decreased during stress more markedly than systolic time. At peak stress 62 of the 161 pts showed reversal of the systolic/diastolic ratio with the duration of systole longer than diastole. In the exercise group, at 100 bpm HR, systolic/diastolic time ratio was lower in the 17 controls (0.74 +/- 0.12) than in patients (0.86 +/- 0.10, p < 0.05 vs. controls). Diastolic filling rate increased from 101 +/- 36 (rest) to 219 +/- 92 ml/m2* s-1 at peak stress (p < 0.5 vs. rest). CONCLUSION: Cardiological systolic and diastolic duration can be monitored during stress by using an acceleration force sensor. Simultaneous calculation of stroke volume allows monitoring diastolic filling rate.Stress-induced "systolic-diastolic mismatch" can be easily quantified and is associated to several cardiac diseases, possibly expanding the spectrum of information obtainable during stress. PMID- 18426560 TI - In the aftermath of teenage suicide: a qualitative study of the psychosocial consequences for the surviving family members. AB - BACKGROUND: Studies of family reactions following teenage suicide are hampered by the psychological difficulties of approaching families and recruiting an unbiased sample of study subjects. By using a small but consecutive series of cases, we examined the qualitative aspects of loosing a teenage family member due to suicide. Such an understanding is important for future organisation of proper programs that provide professional support in the grief process. METHODS: From a large project on teenage unnatural death in northern Sweden 1981-2000 (including 88 suicides), 13 cases from 1995 through 1998 were retrospectively identified and consecutively analysed. Ten families agreed to participate. The open interviews took place 15 to 25 months after the suicide. The information gathered was manually analysed according to a grounded theory model, resulting in allocation of data into one of three domains: post-suicidal reactions, impact on daily living, and families' need for support. RESULTS: Teenager suicide is a devastating trauma for the surviving family and the lack of sustainable explanations for the suicide is a predominant issue in the grief process. The prolonged social and psychological isolation of the families in grief should be challenged. At the time of the interview, the families were still struggling with explaining why the suicide occurred, especially since most suicides had occurred without overt premonitory signs. The bereaved family members were still profoundly affected by the loss, but all had returned to an ostensibly normal life. Post-suicide support was often badly timed and insufficient, especially for younger siblings. CONCLUSION: Family doctors can organise a long-term, individually formulated support scheme for the bereaved, including laymen who can play a most significant role in the grief process. There is also a need for better understanding of the families who have lost a teenager whom committed suicide and for the development and testing of treatment schemes for the bereaved family. PMID- 18426561 TI - Effect of a heated humidifier during continuous positive airway pressure delivered by a helmet. AB - INTRODUCTION: The helmet may be an effective interface for the delivery of noninvasive positive pressure ventilation. The high internal gas volume of the helmet can act as a 'mixing chamber', in which the humidity of the patient's expired alveolar gases increases the humidity of the dry medical gases, thus avoiding the need for active humidification. We evaluated the temperature and humidity of respiratory gases inside the helmet, with and without a heated humidifier, during continuous positive airway pressure (CPAP) delivered with a helmet. METHODS: Nine patients with acute respiratory failure (arterial oxygen tension/fractional inspired oxygen ratio 209 +/- 52 mmHg) and 10 healthy individuals were subjected to CPAP. The CPAP was delivered either through a mechanical ventilator or by continuous low (40 l/min) or high flow (80 l/min). Humidity was measured inside the helmet using a capacitive hygrometer. The level of patient comfort was evaluated using a continuous scale. RESULTS: In patients with acute respiratory failure, the heated humidifier significantly increased the absolute humidity from 18.4 +/- 5.5 mgH2O/l to 34.1 +/- 2.8 mgH2O/l during ventilator CPAP, from 11.4 +/- 4.8 mgH2O/l to 33.9 +/- 1.9 mgH2O/l during continuous low-flow CPAP, and from 6.4 +/- 1.8 mgH2O/l to 24.2 +/- 5.4 mgH2O/l during continuous high-flow CPAP. Without the heated humidifier, the absolute humidity was significantly higher with ventilator CPAP than with continuous low flow and high-flow CPAP. The level of comfort was similar for all the three modes of ventilation and with or without the heated humidifier. The findings in healthy individuals were similar to those in the patients with acute respiratory failure. CONCLUSION: The fresh gas flowing through the helmet with continuous flow CPAP systems limited the possibility to increase the humidity. We suggest that a heated humidifier should be employed with continuous flow CPAP systems. PMID- 18426562 TI - Investigation of the hydrodynamic properties of a new MRI-resistant programmable hydrocephalus shunt. AB - BACKGROUND: The Polaris valve is a newly released hydrocephalus shunt that is designed to drain cerebrospinal fluid (CSF) from the brain ventricles or lumbar CSF space. The aim of this study was to bench test the properties of the Polaris shunt, independently of the manufacturer. METHODS: The Polaris Valve is a ball-on spring valve, which can be adjusted magnetically in vivo. A special mechanism is incorporated to prevent accidental re-adjustment by an external magnetic field. The performance and hydrodynamic properties of the valve were evaluated in the UK Shunt Evaluation Laboratory, Cambridge, UK. RESULTS: The three shunts tested showed good mechanical durability over the 3-month period of testing, and a stable hydrodynamic performance over 45 days. The pressure-flow performance curves, operating, opening and closing pressures were stable. The drainage rate of the shunt increased when a negative outlet pressure (siphoning) was applied. The hydrodynamic parameters fell within the limits specified by the manufacturer and changed according to the five programmed performance levels. Hydrodynamic resistance was dependant on operating pressure, changing from low values of 1.6 mmHg/ml/min at the lowest level to 11.2 mmHg/ml/min at the highest performance level. External programming proved to be easy and reliable. Even very strong magnetic fields (3 Tesla) were not able to change the programming of the valve. However, distortion of magnetic resonance images was present. CONCLUSION: The Polaris Valve is a reliable, adjustable valve. Unlike other adjustable valves (except the Miethke ProGAV valve), the Polaris cannot be accidentally re-adjusted by an external magnetic field. PMID- 18426563 TI - Methylation alterations are not a major cause of PTTG1 misregulation. AB - BACKGROUND: On its physiological cellular context, PTTG1 controls sister chromatid segregation during mitosis. Within its crosstalk to the cellular arrest machinery, relies a checkpoint of integrity for which gained the over name of securin. PTTG1 was found to promote malignant transformation in 3T3 fibroblasts, and further found to be overexpressed in different tumor types. More recently, PTTG1 has been also related to different processes such as DNA repair and found to trans-activate different cellular pathways involving c-myc, bax or p53, among others. PTTG1 over-expression has been correlated to a worse prognosis in thyroid, lung, colorectal cancer patients, and it can not be excluded that this effect may also occur in other tumor types. Despite the clinical relevance and the increasing molecular characterization of PTTG1, the reason for its up regulation remains unclear. METHOD: We analysed PTTG1 differential expression in PC-3, DU-145 and LNCaP tumor cell lines, cultured in the presence of the methyl transferase inhibitor 5-Aza-2'-deoxycytidine. We also tested whether the CpG island mapping PTTG1 proximal promoter evidenced a differential methylation pattern in differentiated thyroid cancer biopsies concordant to their PTTG1 immunohistochemistry status. Finally, we performed whole-genome LOH studies using Affymetix 50 K microarray technology and FRET analysis to search for allelic imbalances comprising the PTTG1 locus. CONCLUSION: Our data suggest that neither methylation alterations nor LOH are involved in PTTG1 over-expression. These data, together with those previously reported, point towards a post transcriptional level of misregulation associated to PTTG1 over-expression. PMID- 18426564 TI - Carpal tunnel syndrome and the "double crush" hypothesis: a review and implications for chiropractic. AB - Upton and McComas claimed that most patients with carpal tunnel syndrome not only have compressive lesions at the wrist, but also show evidence of damage to cervical nerve roots. This "double crush" hypothesis has gained some popularity among chiropractors because it seems to provide a rationale for adjusting the cervical spine in treating carpal tunnel syndrome. Here I examine use of the concept by chiropractors, summarize findings from the literature, and critique several studies aimed at supporting or refuting the hypothesis. Although the hypothesis also has been applied to nerve compressions other than those leading to carpal tunnel syndrome, this discussion mainly examines the original application - "double crush" involving both cervical spinal nerve roots and the carpal tunnel. I consider several categories: experiments to create double crush syndrome in animals, case reports, literature reviews, and alternatives to the original hypothesis. A significant percentage of patients with carpal tunnel syndrome also have neck pain or cervical nerve root compression, but the relationship has not been definitively explained. The original hypothesis remains controversial and is probably not valid, at least for sensory disturbances, in carpal tunnel syndrome. However, even if the original hypothesis is importantly flawed, evaluation of multiple sites still may be valuable. The chiropractic profession should develop theoretical models to relate cervical dysfunction to carpal tunnel syndrome, and might incorporate some alternatives to the original hypothesis. I intend this review as a starting point for practitioners, educators, and students wishing to advance chiropractic concepts in this area. PMID- 18426565 TI - Recommendations by Cochrane Review Groups for assessment of the risk of bias in studies. AB - BACKGROUND: Assessing the risk of bias in individual studies in a systematic review can be done using individual components or by summarizing the study quality in an overall score. METHODS: We examined the instructions to authors of the 50 Cochrane Review Groups that focus on clinical interventions for recommendations on methodological quality assessment of studies. RESULTS: Forty one of the review groups (82%) recommended quality assessment using components and nine using a scale. All groups recommending components recommended to assess concealment of allocation, compared to only two of the groups recommending scales (P < 0.0001). Thirty-five groups (70%) recommended assessment of sequence generation and 21 groups (42%) recommended assessment of intention-to-treat analysis. Only 28 groups (56%) had specific recommendations for using the quality assessment of studies analytically in reviews, with sensitivity analysis, quality as an inclusion threshold and subgroup analysis being the most commonly recommended methods. The scales recommended had problems in the individual items and some of the groups recommending components recommended items not related to bias in their quality assessment. CONCLUSION: We found that recommendations by some groups were not based on empirical evidence and many groups had no recommendations on how to use the quality assessment in reviews. We suggest that all Cochrane Review Groups refer to the Cochrane Handbook for Systematic Reviews of Interventions, which is evidence-based, in their instructions to authors and that their own guidelines are kept to a minimum and describe only how methodological topics that are specific to their fields should be handled. PMID- 18426566 TI - Paraneoplastic cerebellar degeneration as a presentation of breast cancer - a case report and review of the literature. AB - Paraneoplastic cerebellar degeneration is part of a rare spectrum of neurological syndromes whereby gynaecological, lung or breast cancers present primarily with neurological manifestations. The presence of onconeural antibodies and PET scanning help in the challenging diagnosis of these conditions but despite the treatment of the primary cancer, the prognosis for the neurological symptoms is poor. PMID- 18426567 TI - Study protocol of the German "Registry for the detection of late sequelae after radiotherapy in childhood and adolescence" (RiSK). AB - BACKGROUND: Late effects after radiotherapy in childhood and adolescence have mainly been characterized retrospectively with small patient numbers. However, these analyses are limited due to little information regarding organ dose levels in many cases. To overcome this limitation, the German Group of Paediatric Radiation Oncology (APRO) established the "Registry for the evaluation of late side effects after radiation in childhood and adolescence" (RiSK). The study protocol and the documentation forms are given in this publication. METHODS/DESIGN: Radiation parameters including detailed organ doses as well as toxicity evaluations are collected prospectively from centres all over Germany. Standardized documentation forms are used. These forms are given in an English and German version as additional files to this publication. Documentation is planned for all children who receive radiotherapy in one of the therapy trials of the "German Society of Paediatric Oncology and Haematology (GPOH)". The study started in a pilot phase in June 2001 in few centres. Since 2004 documentation has been performed all over Germany and is still on-going. DISCUSSION: To our knowledge, "RiSK" is the only multi-centre study that evaluates radiation associated side effects prospectively with detailed information about organ dose levels. With ongoing recruitment and prolongation of follow-up powerful data will be obtained in a few years. A broad use and international cooperation are welcome. PMID- 18426569 TI - A rare cause of forearm pain: anterior branch of the medial antebrachial cutaneous nerve injury: a case report. AB - INTRODUCTION: Medial antebrachial cutaneous nerve (MACN) neuropathy is reported to be caused by iatrogenic reasons. Although the cases describing the posterior branch of MACN neuropathy are abundant, only one case caused by lipoma has been found to describe the anterior branch of MACN neuropathy in the literature. As for the reason for the forearm pain, we report the only case describing isolated anterior branch of MACN neuropathy which has developed due to repeated minor trauma. CASE PRESENTATION: We report a 37-year-old woman patient with pain in her medial forearm and elbow following the shaking of a rug. Pain and symptoms of dysestesia in the distribution of the right MACN were found. Electrophysiological examination confirmed the normality of the main nerve trunks of the right upper limb and demonstrated abnormalities of the right MACN when compared with the left side. Sensory action potential (SAP) amplitude on the right anterior branch of the MACN was detected to be lower in proportion to the left. In the light of these findings, NSAI drug and physical therapy was performed. Dysestesia and pain were relieved and no recurrence was observed after a follow-up of 14 months. CONCLUSION: MACN neuropathy should be taken into account for the differential diagnosis of the patients with complaints of pain and dysestesia in medial forearm and anteromedial aspect of the elbow. PMID- 18426568 TI - Psychological, social and biological determinants of ill health (pSoBid): study protocol of a population-based study. AB - BACKGROUND: Disadvantaged communities suffer higher levels of physical and mental ill health than more advantaged communities. The purpose of the present study was to examine the psychosocial, behavioural and biological determinants of ill health within population groups in Glasgow that differed in socioeconomic status and in their propensity to develop chronic disease especially coronary heart disease and Type 2 diabetes mellitus. METHODS: Participants were selected at random from areas known to be at the extremes of the socioeconomic continuum in Glasgow. Within the categories of least deprived and most deprived, recruitment was stratified by sex and age to achieve an overall sample containing approximately equal numbers of males and females and an even distribution across the age categories 35-44, 45-54 and 55-64 years. Individuals were invited by letter to attend for assessment of their medical history, risk factor status, cognitive function and psychological profile, morbidity, and carotid intima-media thickness and plaque count as indices of atherosclerosis. Anonymised data on study subjects were collected from the General Practice Administration System for Scotland to analyse characteristics of participants and non-participants. RESULTS: 700 subjects were recruited. The response (active participants per 100 invitation letters) in the least deprived group was 35.1% and in the most deprived group was 20.3%. Lowest response was seen in young males (least deprived 22.4% and most deprived 14.1%). CONCLUSION: This cross-sectional study recruited the planned sample of subjects from least deprived and most deprived areas within Glasgow. As evident in other studies response differed between the most and least deprived areas. This study brought together researchers/academics from diverse disciplines to build a more sophisticated understanding of the determinants of health inequalities than can be achieved through unidisciplinary approaches. Future analyses will enable an understanding of the relationships between the different types of measure, and of the pathways that link poverty, biology, behaviour and psychology and lead to health inequalities. PMID- 18426570 TI - Endotoxin receptor CD14 in PiZ alpha-1-antitrypsin deficiency individuals. AB - BACKGROUND: CD14, a receptor for lipopolysaccharides (LPS), is found in both a membrane-bound form (mCD14) and a soluble form (sCD14). It is suggested that sCD14 is mainly released from blood monocytes by serine protease-mediated shedding. Because alpha1-antitrypsin (AAT), an inhibitor of serine proteases, has been shown to regulate CD14 expression in human monocytes in vitro, we sought to investigate plasma levels of sCD14 and monocyte expression of mCD14 in subjects at age 30 years with normal MM and deficient PiZZ and PiSZ genotypes of AAT. METHODS: Plasma levels of AAT and sCD14 were measured in 75 PiZZ and 34 PiSZ individuals with normal lung function identified from the Swedish neonatal AAT deficiency screening, and in 95 age matched PiMM controls. The mCD14 expression in monocytes from 9 PiZZ, 6 PiSZ and 11 PiMM subjects was analysed by FACS and Quantitative Real Time Reverse Transcription PCA. RESULTS: As expected, plasma AAT concentrations were PiMM>PiSZ>PiZZ (p < 0.001). Plasma sCD14 levels were higher in PiZZ than in PiMM subjects (p < 0.01). The expression level of mCD14 was higher (1.89-fold) in monocytes isolated from PiZZ subjects compared to PiMM controls (p = 0.00189). CONCLUSION: This study is the first to show higher levels of plasma sCD14 and monocyte mCD14 expression in young, clinically healthy PiZZ AAT subjects. PMID- 18426571 TI - Effects of an attention demanding task on dynamic stability during treadmill walking. AB - BACKGROUND: People exhibit increased difficulty balancing when they perform secondary attention-distracting tasks while walking. However, a previous study by Grabiner and Troy (J. Neuroengineering Rehabil., 2005) found that young healthy subjects performing a concurrent Stroop task while walking on a motorized treadmill exhibited decreased step width variability. However, measures of variability do not directly quantify how a system responds to perturbations. This study re-analyzed data from Grabiner and Troy 2005 to determine if performing the concurrent Stroop task directly affected the dynamic stability of walking in these same subjects. METHODS: Thirteen healthy volunteers walked on a motorized treadmill at their self-selected constant speed for 10 minutes both while performing the Stroop test and during undisturbed walking. This Stroop test consisted of projecting images of the name of one color, printed in text of a different color, onto a wall and asking subjects to verbally identify the color of the text. Three-dimensional motions of a marker attached to the base of the neck (C5/T1) were recorded. Marker velocities were calculated over 3 equal intervals of 200 sec each in each direction. Mean variability was calculated for each time series as the average standard deviation across all strides. Both "local" and "orbital" dynamic stability were quantified for each time series using previously established methods. These measures directly quantify how quickly small perturbations grow or decay, either continuously in real time (local) or discretely from one cycle to the next (orbital). Differences between Stroop and Control trials were evaluated using a 2-factor repeated measures ANOVA. RESULTS: Mean variability of trunk movements was significantly reduced during the Stroop tests compared to normal walking. Conversely, local and orbital stability results were mixed: some measures showed slight increases, while others showed slight decreases. In many cases, different subjects responded differently to the Stroop test. While some of our comparisons reached statistical significance, many did not. In general, measures of variability and dynamic stability reflected different properties of walking dynamics, consistent with previous findings. CONCLUSION: These findings demonstrate that the decreased movement variability associated with the Stroop task did not translate to greater dynamic stability. PMID- 18426572 TI - Cecal diverticulitis mimicking acute Appendicitis: a report of 4 cases. AB - Diverticulum of the cecum is a rare, benign, generally asymptomatic lesion that manifests itself only following inflammatory or hemorrhagic complications. Most patients with inflammation of a solitary diverticulum of the cecum present with abdominal pain that is indistinguishable from acute appendicitis. The optimal management of this condition is still controversial, ranging from conservative antibiotic treatment to aggressive resection. We describe four cases that presented with symptoms suggestive of appendicitis, but were found at operation to have an inflamed solitary diverticulum. PMID- 18426573 TI - Calcium pyrophosphate dihydrate deposition disease (CPPD)/Pseudogout of the temporomandibular joint - FNA findings and microanalysis. AB - We report a case of a Calcium pyrophosphate dihydrate deposition disease (CPPD) presenting as a mass in the parotid and temporomandibular joint (TMJ) that simulated a parotid tumor. A 35 year-old man presented with pain in the left ear area. A CT Scan of the area showed a large, calcified mass surrounding the left condylar head, and extending into the infratemporal fossa. FNA of the mass showed birefringent crystals, most of which were rhomboid with occasional ones being needle shaped, embedded in an amorphous pink substance. Scanning electron microscopy (SEM) with energy dispersive x-ray spectroscopy (EDS) of these crystals showed peaks corresponding to calcium and phosphorus. SEM/EDS is a rapid method of diagnosing calcium pyrophosphate dihydrate deposition disease (CPPD) and an alternative to more commonly used method of special staining of cell block sections coupled with polarizing microscopy. PMID- 18426574 TI - Myoepithelial cells: any role in aspiration cytology smears of breast tumors? AB - AIMS AND OBJECTIVE: To study the role of myoepithelial (ME) cells in distinguishing benign, proliferative breast diseases (PBD) and frank malignant breast lesions. MATERIALS AND METHODS: In this study, histology proven 71 cases of fine needle aspiration cytology (FNAC) of palpable breast lesions were selected. There were 30 invasive carcinomas (24 infiltrating duct carcinoma and 6 infiltrating lobular carcinoma), 25 cases of benign lesion (21 fibroadenomas and 4 fibrocystic lesions) and 11 proliferative breast diseases (other than carcinoma in situ) and five cases of carcinoma in situ. The number of ME cells were estimated in respect to 1000 ductal cells. In every case at least 20 high power fields (x 40) were studied. Quantitative estimation of ME cell was correlated with the final diagnosis. Corresponding histopathology cases were also evaluated for diagnostic confirmation along with the pattern of distribution of ME cells. The ME cells were also quantitated on histopathology sections on smooth muscle actin (SMA) immunostained sections. RESULTS: The mean number of ME cells per 1000 ductal cells on cytology smears was 5.1 +/- 5.5, 30.8 +/- 25, 28.3 +/- 20.2, and 38.4 +/- 38.8 in malignant, carcinoma in situ, PBD and benign breast lesions respectively. The non parametric Mann Whitney test showed significant difference in number of the ME cells between benign and malignant groups (p < .000), PBD and malignant groups (p < .000) and carcinoma in situ and malignant group (p < .001). However, it was insignificant between benign and PBD group, and PBD and carcinoma in situ (p > .01). In SMA stained histopathology sections, ME cell in benign, PBD, carcinoma in situ and malignant cases were 741.12 +/- 248, 238 +/- 172, 121.6 +/- 115 and 15.6 +/- 25.1 respectively. Statistical analysis showed significantly different number of ME cell between benign versus PBD group, carcinoma in situ and malignant group. It was also significant between PBD versus malignant, and carcinoma in situ versus malignant (p < .001, Mann Whitney test). However number of ME cell was not significant between PBD versus carcinoma in situ. CONCLUSION: The number of ME cell in breast lesions may be helpful in distinguishing PBD versus invasive malignant tumors on FNAC smears. However it is not helpful to distinguish benign lesions versus PBD. PMID- 18426576 TI - Practical aspects in the management of hypokalemic periodic paralysis. AB - Management considerations in hypokalemic periodic paralysis include accurate diagnosis, potassium dosage for acute attacks, choice of diuretic for prophylaxis, identification of triggers, creating a safe physical environment, peri-operative measures, and issues in pregnancy. A positive genetic test in the context of symptoms is the gold standard for diagnosis. Potassium chloride is the favored potassium salt given at 0.5-1.0 mEq/kg for acute attacks. The oral route is favored, but if necessary, a mannitol solvent can be used for intravenous administration. Avoidance of or potassium prophylaxis for common triggers, such as rest after exercise, high carbohydrate meals, and sodium, can prevent attacks. Chronically, acetazolamide, dichlorphenamide, or potassium-sparing diuretics decrease attack frequency and severity but are of little value acutely. Potassium, water, and a telephone should always be at a patient's bedside, regardless of the presence of weakness. Perioperatively, the patient's clinical status should be checked frequently. Firm data on the management of periodic paralysis during pregnancy is lacking. Patient support can be found at http://www.periodicparalysis.org. PMID- 18426575 TI - Capturing doping attitudes by self-report declarations and implicit assessment: a methodology study. AB - BACKGROUND: Understanding athletes' attitudes and behavioural intentions towards performance enhancement is critical to informing anti-doping intervention strategies. Capturing the complexity of these attitudes beyond verbal declarations requires indirect methods. This pilot study was aimed at developing and validating a method to assess implicit doping attitudes using an Implicit Associations Test (IAT) approach. METHODS: The conventional IAT evaluation task (categorising 'good' and 'bad' words) was combined with a novel 'doping' versus 'nutrition supplements' category pair to create a performance-enhancement related IAT protocol (PE-IAT). The difference between average response times to 'good doping' and 'bad-doping' combinations represents an estimate of implicit attitude towards doping in relation to nutritional supplements. 111 sports and exercise science undergraduates completed the PE-IAT, the Performance Enhancement Attitude Scale (PEAS) and answered questions regarding their beliefs about doping. RESULTS: Longer response times were observed in the mixed category discrimination trials where categories 'good' and 'doping' shared the same response key (compared to 'bad-doping' combination on the same key) indicating a less favourable evaluation of doping substances. The PE-IAT measure did not correlate significantly with the declared doping attitudes (r = .181, p = .142), indicating a predictable partial dissociation. Action-oriented self-report expressed stronger associations with PE-IAT: participants who declared they would consider using doping showed significantly less implicit negativity towards banned substances (U = 109.00, p = .047). Similarly, those who reported more lenient explicit attitudes towards doping or expressly supported legalizing it, showed less implicit negativity towards doping in the sample, although neither observed differences reached statistical significance (t = 1.300, p = .198, and U = 231.00, p = .319, respectively). Known-group validation strategy yielded mixed results: while competitive sport participants scored significantly lower than non competitive ones on the PEAS (t = -2.71, p = .008), the two groups did not differ on PE-IAT (t = -.093, p = .926). CONCLUSION: The results suggest a potential of the PE-IAT method to capture undeclared attitudes to doping and predict behaviour, which can support targeted anti-doping intervention and related research. The initial evidence of validity is promising but also indicates a need for improvement to the protocol and stimulus material. PMID- 18426577 TI - Effect of a proprietary Magnolia and Phellodendron extract on stress levels in healthy women: a pilot, double-blind, placebo-controlled clinical trial. AB - BACKGROUND: Recent research has established correlations between stress, anxiety, insomnia and excess body weight and these correlations have significant implications for health. This study measured the effects of a proprietary blend of extracts of Magnolia officinalis and Phellodendron amurense (Relora) on anxiety, stress and sleep in healthy premenopausal women. METHODS: This randomized, parallel, placebo controlled clinical study was conducted with healthy, overweight (BMI 25 to 34.9), premenopausal female adults, between the ages of 20 and 50 years, who typically eat more in response to stressful situations and scores above the national mean for women on self-reporting anxiety. The intervention was Relora (250 mg capsules) or identical placebo 3 times daily for 6 weeks. Anxiety as measured by the Spielberger STATE-TRAIT questionnaires, salivary amylase and cortisol levels, Likert Scales/Visual Analog Scores for sleep quality and latency, appetite, and clinical markers of safety. The study was conducted by Miami Research Associates, a clinical research organization in Miami, FL. RESULTS: The intent-to-treat population consisted of 40 subjects with 26 participants completing the study. There were no significant adverse events. Relora was effective, in comparison to placebo, in reducing temporary, transitory anxiety as measured by the Spielberger STATE anxiety questionnaire. It was not effective in reducing long-standing feelings of anxiety or depression as measured using the Spielberger TRAIT questionnaire. Other assessments conducted in this study including salivary cortisol and amylase levels, appetite, body morphology and sleep quality/latency were not significantly changed by Relora in comparison to placebo. CONCLUSION: This pilot study indicates that Relora may offer some relief for premenopausal women experiencing mild transitory anxiety. There were no safety concerns or significant adverse events observed in this study. PMID- 18426578 TI - DNA watermarks: a proof of concept. AB - BACKGROUND: DNA-based watermarks are helpful tools to identify the unauthorized use of genetically modified organisms (GMOs) protected by patents. In silico analyses showed that in coding regions synonymous codons can be used to insert encrypted information into the genome of living organisms by using the DNA-Crypt algorithm. RESULTS: We integrated an authenticating watermark in the Vam7 sequence. For our investigations we used a mutant Saccharomyces cerevisiae strain, called CG783, which has an amber mutation within the Vam7 sequence. The CG783 cells are unable to sporulate and in addition display an abnormal vacuolar morphology. Transformation of CG783 with pRS314 Vam7 leads to a phenotype very similar to the wildtype yeast strain CG781. The integrated watermark did not influence the function of Vam7 and the resulting phenotype of the CG783 cells transformed with pRS314 Vam7-TB shows no significant differences compared to the CG783 cells transformed with pRS314 Vam7. CONCLUSION: From our experiments we conclude that the DNA watermarks produced by DNA-Crypt do not influence the translation from mRNA into protein. By analyzing the vacuolar morphology, growth rate and ability to sporulate we confirmed that the resulting Vam7 protein was functionally active. PMID- 18426579 TI - Cathepsin D SNP associated with increased risk of variant Creutzfeldt-Jakob disease. AB - BACKGROUND: Variant Creutzfeldt-Jakob disease (vCJD) originally resulted from the consumption of foodstuffs contaminated by bovine spongiform encephalopathy (BSE) material, with 163 confirmed cases in the UK to date. Many thousands are likely to have been exposed to dietary infection and so it is important (for surveillance, epidemic modelling, public health and understanding pathogenesis) to identify genetic factors that may affect individual susceptibility to infection. This study looked at a polymorphism in the cathepsin D gene (refSNP ID: rs17571) previously examined in Alzheimer's disease (AD). METHODS: Blood samples taken from 110 vCJD patients were tested for the C-T base change, and genotype data were compared with published frequencies for a control population using multiple logistic regression. RESULTS: There was a significant excess of the cathepsin D polymorphism TT genotype in the vCJD cohort compared to controls. The TT genotype was found to have a 9.75 fold increase in risk of vCJD compared to the CT genotype and a 10.92 fold increase compared to the CC genotype. CONCLUSION: This mutation event has been observed to alter the protease activity of the cathepsin D protein and has been linked to an increase in amyloid beta plaque formation in AD. vCJD neuropathology is characterised by the presence of amyloid plaques, formed from the prion protein, and therefore alterations in the amyloid processing activity of cathepsin D may affect the neuropathogenesis of this disease. PMID- 18426580 TI - Network motif-based identification of transcription factor-target gene relationships by integrating multi-source biological data. AB - BACKGROUND: Integrating data from multiple global assays and curated databases is essential to understand the spatio-temporal interactions within cells. Different experiments measure cellular processes at various widths and depths, while databases contain biological information based on established facts or published data. Integrating these complementary datasets helps infer a mutually consistent transcriptional regulatory network (TRN) with strong similarity to the structure of the underlying genetic regulatory modules. Decomposing the TRN into a small set of recurring regulatory patterns, called network motifs (NM), facilitates the inference. Identifying NMs defined by specific transcription factors (TF) establishes the framework structure of a TRN and allows the inference of TF target gene relationship. This paper introduces a computational framework for utilizing data from multiple sources to infer TF-target gene relationships on the basis of NMs. The data include time course gene expression profiles, genome-wide location analysis data, binding sequence data, and gene ontology (GO) information. RESULTS: The proposed computational framework was tested using gene expression data associated with cell cycle progression in yeast. Among 800 cell cycle related genes, 85 were identified as candidate TFs and classified into four previously defined NMs. The NMs for a subset of TFs are obtained from literature. Support vector machine (SVM) classifiers were used to estimate NMs for the remaining TFs. The potential downstream target genes for the TFs were clustered into 34 biologically significant groups. The relationships between TFs and potential target gene clusters were examined by training recurrent neural networks whose topologies mimic the NMs to which the TFs are classified. The identified relationships between TFs and gene clusters were evaluated using the following biological validation and statistical analyses: (1) Gene set enrichment analysis (GSEA) to evaluate the clustering results; (2) Leave-one-out cross validation (LOOCV) to ensure that the SVM classifiers assign TFs to NM categories with high confidence; (3) Binding site enrichment analysis (BSEA) to determine enrichment of the gene clusters for the cognate binding sites of their predicted TFs; (4) Comparison with previously reported results in the literatures to confirm the inferred regulations. CONCLUSION: The major contribution of this study is the development of a computational framework to assist the inference of TRN by integrating heterogeneous data from multiple sources and by decomposing a TRN into NM-based modules. The inference capability of the proposed framework is verified statistically (e.g., LOOCV) and biologically (e.g., GSEA, BSEA, and literature validation). The proposed framework is useful for inferring small NM based modules of TF-target gene relationships that can serve as a basis for generating new testable hypotheses. PMID- 18426581 TI - Estimated pre-morbid IQ effects on cognitive and functional outcomes in Alzheimer disease: a longitudinal study in a treated cohort. AB - BACKGROUND: Cognitive reserve is thought to influence the degree of neuropathology needed for diagnosis of Alzheimer disease (AD). Cognitive reserve can be operationally defined as the hypothesized capacity of the mature adult brain to sustain the effects of disease or injury without manifesting clinical symptoms of AD, but sufficient to cause clinical dementia in an individual possessing less cognitive reserve. Its effect on the subsequent course of AD is less clear. Pre-morbid IQ is a useful measure of cognitive reserve. METHODS: We studied 659 consecutive patients with AD at a tertiary referral memory clinic. Patients were assessed on six cognitive tests at baseline. Activities of Daily Living (ADL) were measured on the Instrumental Activities of Daily Living (IADL) scale and Physical Self-Maintenance Scale (PSMS). The National Adult Reading Test (NART) was used to estimate pre-morbid IQ. Patients were followed up after starting a cholinesterase inhibitor over 78 weeks. Mixed general linear models estimated the effects of NART on cognition and ADL. RESULTS: Three hundred and fifty-five patients had NART scored with a mean estimated pre-morbid IQ of 104.7 (standard deviation 18.5). NART increased overall cognitive ability by 2.7% for every 10 IQ points (p < .001). There was a trend for an effect on the change in cognition over time (p = .065) with higher NART associated with improvement of cognitive ability over time. After adjusting for age and sex, a 10 point increase in NART was associated with an improvement of 2% in ADL scores, but this effect was explained by NART's influence on contemporaneous cognitive ability. CONCLUSION: Our data support the hypothesis that cognitive reserve continues to have a limited influence on cognition after AD has been diagnosed and thus, indirectly, has an impact on ADL. PMID- 18426582 TI - Percent body fat estimations in college men using field and laboratory methods: a three-compartment model approach. AB - BACKGROUND: Methods used to estimate percent body fat can be classified as a laboratory or field technique. However, the validity of these methods compared to multiple-compartment models has not been fully established. The purpose of this study was to determine the validity of field and laboratory methods for estimating percent fat (%fat) in healthy college-age men compared to the Siri three-compartment model (3C). METHODS: Thirty-one Caucasian men (22.5 +/- 2.7 yrs; 175.6 +/- 6.3 cm; 76.4 +/- 10.3 kg) had their %fat estimated by bioelectrical impedance analysis (BIA) using the BodyGram computer program (BIA AK) and population-specific equation (BIA-Lohman), near-infrared interactance (NIR) (Futrex(R) 6100/XL), four circumference-based military equations [Marine Corps (MC), Navy and Air Force (NAF), Army (A), and Friedl], air-displacement plethysmography (BP), and hydrostatic weighing (HW). RESULTS: All circumference based military equations (MC = 4.7% fat, NAF = 5.2% fat, A = 4.7% fat, Friedl = 4.7% fat) along with NIR (NIR = 5.1% fat) produced an unacceptable total error (TE). Both laboratory methods produced acceptable TE values (HW = 2.5% fat; BP = 2.7% fat). The BIA-AK, and BIA-Lohman field methods produced acceptable TE values (2.1% fat). A significant difference was observed for the MC and NAF equations compared to both the 3C model and HW (p < 0.006). CONCLUSION: Results indicate that the BP and HW are valid laboratory methods when compared to the 3C model to estimate %fat in college-age Caucasian men. When the use of a laboratory method is not feasible, BIA-AK, and BIA-Lohman are acceptable field methods to estimate %fat in this population. PMID- 18426583 TI - Esophageal squamous cell carcinoma presenting with extensive skin lesions: a case report. AB - INTRODUCTION: Esophageal squamous cell carcinoma (ESCC) is the most common histological subtype of cancer in the upper and middle esophagus and is characterized by a high rate of mortality. The incidence of esophageal cancer varies greatly among regions of the world and occurs at a high frequency in Asia and South America. CASE PRESENTATION: In our department, a 51-year-old man was diagnosed with ESCC after presenting with extensive disseminated skin nodules. Biopsy of the nodules showed metastatic ESCC. Cutaneous manifestations of esophageal neoplasia are very rare and are mainly described for esophageal adenocarcinoma (EADC). Here we report a very uncommon case of extensive skin metastases of ESCC. CONCLUSION: Early biopsies of suspicious skin lesions are important and should be performed in patients with unclear symptoms such as weight loss or dysphagia and especially in patients with a history of cancer, since they can reveal the existence of a distant malignant disease leading to diagnosis and prompt therapy. PMID- 18426584 TI - Phylogenomics of plant genomes: a methodology for genome-wide searches for orthologs in plants. AB - BACKGROUND: Gene ortholog identification is now a major objective for mining the increasing amount of sequence data generated by complete or partial genome sequencing projects. Comparative and functional genomics urgently need a method for ortholog detection to reduce gene function inference and to aid in the identification of conserved or divergent genetic pathways between several species. As gene functions change during evolution, reconstructing the evolutionary history of genes should be a more accurate way to differentiate orthologs from paralogs. Phylogenomics takes into account phylogenetic information from high-throughput genome annotation and is the most straightforward way to infer orthologs. However, procedures for automatic detection of orthologs are still scarce and suffer from several limitations. RESULTS: We developed a procedure for ortholog prediction between Oryza sativa and Arabidopsis thaliana. Firstly, we established an efficient method to cluster A. thaliana and O. sativa full proteomes into gene families. Then, we developed an optimized phylogenomics pipeline for ortholog inference. We validated the full procedure using test sets of orthologs and paralogs to demonstrate that our method outperforms pairwise methods for ortholog predictions. CONCLUSION: Our procedure achieved a high level of accuracy in predicting ortholog and paralog relationships. Phylogenomic predictions for all validated gene families in both species were easily achieved and we can conclude that our methodology outperforms similarly based methods. PMID- 18426585 TI - A spatial dissection of the Arabidopsis floral transcriptome by MPSS. AB - BACKGROUND: We have further characterized floral organ-localized gene expression in the inflorescence of Arabidopsis thaliana by comparison of massively parallel signature sequencing (MPSS) data. Six libraries of RNA sequence tags from immature inflorescence tissues were constructed and matched to their respective loci in the annotated Arabidopsis genome. These signature libraries survey the floral transcriptome of wild-type tissue as well as the floral homeotic mutants, apetala1, apetala3, agamous, a superman/apetala1 double mutant, and differentiated ovules dissected from the gynoecia of wild-type inflorescences. Comparing and contrasting these MPSS floral expression libraries enabled demarcation of transcripts enriched in the petals, stamens, stigma-style, gynoecia, and those with predicted enrichment within the sepal/sepal-petals, petal-stamens, or gynoecia-stamens. RESULTS: By comparison of expression libraries, a total of 572 genes were found to have organ-enriched expression within the inflorescence. The bulk of characterized organ-enriched transcript diversity was noted in the gynoecia and stamens, whereas fewer genes demonstrated sepal or petal-localized expression. Validation of the computational analyses was performed by comparison with previously published expression data, in situ hybridizations, promoter-reporter fusions, and reverse transcription PCR. A number of well-characterized genes were accurately delineated within our system of transcript filtration. Moreover, empirical validations confirm MPSS predictions for several genes with previously uncharacterized expression patterns. CONCLUSION: This extensive MPSS analysis confirms and supplements prior microarray floral expression studies and illustrates the utility of sequence survey-based expression analysis in functional genomics. Spatial floral expression data accrued by MPSS and similar methods will be advantageous in the elucidation of more comprehensive genetic regulatory networks governing floral development. PMID- 18426586 TI - Minimal nutrition intervention with high-protein/low-carbohydrate and low-fat, nutrient-dense food supplement improves body composition and exercise benefits in overweight adults: A randomized controlled trial. AB - BACKGROUND: Exercise and high-protein/reduced-carbohydrate and -fat diets have each been shown separately, or in combination with an energy-restricted diet to improve body composition and health in sedentary, overweight (BMI > 25) adults. The current study, instead, examined the physiological response to 10 weeks of combined aerobic and resistance exercise (EX) versus exercise + minimal nutrition intervention designed to alter the macronutrient profile, in the absence of energy restriction, using a commercially available high-protein/low-carbohydrate and low-fat, nutrient-dense food supplement (EXFS); versus control (CON). METHODS: Thirty-eight previously sedentary, overweight subjects (female = 19; male = 19) were randomly assigned to either CON (n = 10), EX (n = 14) or EXFS (n = 14). EX and EXFS participated in supervised resistance and endurance training (2x and 3x/wk, respectively); EXFS consumed 1 shake/d (weeks 1 and 2) and 2 shakes/d (weeks 3-10). RESULTS: EXFS significantly decreased total energy, carbohydrate and fat intake (-14.4%, -27.2% and -26.7%, respectively; p < 0.017), and increased protein and fiber intake (+52.1% and +21.2%, respectively; p < 0.017). EX and EXFS significantly decreased fat mass (-4.6% and -9.3%, respectively; p < 0.017), with a greater (p < 0.05) decrease in EXFS than EX and CON. Muscle mass increase only reached significance in EXFS (+2.3%; p < 0.017), which was greater (p < 0.05) than CON but not EX (+1.1%). Relative VO2max improved in both exercise groups (EX = +5.0% and EXFS = +7.9%; p < 0.017); however, only EXFS significantly improved absolute VO2max (+6.2%; p = 0.001). Time-to-exhaustion during treadmill testing increased in EX (+9.8%) but was significantly less (p < 0.05) than in EXFS (+21.2%). Total cholesterol and LDL decreased only in the EXFS (-12.0% and -13.3%, respectively; p < 0.017). Total cholesterol-to-HDL ratio, however, decreased significantly (p < 0.017) in both exercise groups. CONCLUSION: Absent energy restriction or other dietary controls, provision of a high-protein/low-carbohydrate and -fat, nutrient-dense food supplement significantly, 1) modified ad libitum macronutrient and energy intake (behavior effect), 2) improved physiological adaptations to exercise (metabolic advantage), and 3) reduced the variability of individual responses for fat mass, muscle mass and time-to-exhaustion - all three variables improving in 100% of EXFS subjects. PMID- 18426587 TI - The brown adipocyte differentiation pathway in birds: an evolutionary road not taken. AB - BACKGROUND: Thermogenic brown adipose tissue has never been described in birds or other non-mammalian vertebrates. Brown adipocytes in mammals are distinguished from the more common white fat adipocytes by having numerous small lipid droplets rather than a single large one, elevated numbers of mitochondria, and mitochondrial expression of the nuclear gene UCP1, the uncoupler of oxidative phosphorylation responsible for non-shivering thermogenesis. RESULTS: We have identified in vitro inductive conditions in which mesenchymal cells isolated from the embryonic chicken limb bud differentiate into avian brown adipocyte-like cells (ABALCs) with the morphological and many of the biochemical properties of terminally differentiated brown adipocytes. Avian, and as we show here, lizard species lack the gene for UCP1, although it is present in amphibian and fish species. While ABALCs are therefore not functional brown adipocytes, they are generated by a developmental pathway virtually identical to brown fat differentiation in mammals: both the common adipogenic transcription factor peroxisome proliferator-activated receptor-gamma (PPARgamma), and a coactivator of that factor specific to brown fat differentiation in mammals, PGC1alpha, are elevated in expression, as are mitochondrial volume and DNA. Furthermore, ABALCs induction resulted in strong transcription from a transfected mouse UCP1 promoter. CONCLUSION: These findings strongly suggest that the brown fat differentiation pathway evolved in a common ancestor of birds and mammals and its thermogenicity was lost in the avian lineage, with the degradation of UCP1, after it separated from the mammalian lineage. Since this event occurred no later than the saurian ancestor of birds and lizards, an implication of this is that dinosaurs had neither UCP1 nor canonically thermogenic brown fat. PMID- 18426588 TI - Major copy proportion analysis of tumor samples using SNP arrays. AB - BACKGROUND: Single nucleotide polymorphisms (SNPs) are the most common genetic variations in the human genome and are useful as genomic markers. Oligonucleotide SNP microarrays have been developed for high-throughput genotyping of up to 900,000 human SNPs and have been used widely in linkage and cancer genomics studies. We have previously used Hidden Markov Models (HMM) to analyze SNP array data for inferring copy numbers and loss-of-heterozygosity (LOH) from paired normal and tumor samples and unpaired tumor samples. RESULTS: We proposed and implemented major copy proportion (MCP) analysis of oligonucleotide SNP array data. A HMM was constructed to infer unobserved MCP states from observed allele specific signals through emission and transition distributions. We used 10 K, 100 K and 250 K SNP array datasets to compare MCP analysis with LOH and copy number analysis, and showed that MCP performs better than LOH analysis for allelic imbalanced chromosome regions and normal contaminated samples. The major and minor copy alleles can also be inferred from allelic-imbalanced regions by MCP analysis. CONCLUSION: MCP extends tumor LOH analysis to allelic imbalance analysis and supplies complementary information to total copy numbers. MCP analysis of mixing normal and tumor samples suggests the utility of MCP analysis of normal-contaminated tumor samples. The described analysis and visualization methods are readily available in the user-friendly dChip software. PMID- 18426589 TI - Characterisation of human papillomavirus (HPV) genotypes in the Azorean population, Terceira island. AB - BACKGROUND: Human papillomavirus detection is very important for the evaluation of prevention strategies in cervical cancer. In the Azorean population, the virus prevalence has never been studied, and there is no data available to preview a successful outcome with HPV vaccination. In this article, our objective is to characterise the HPV genotypes in Terceira Island, contributing for the epidemiological knowledge on the virus infection. RESULTS: Cervical samples were collected from 289 women aged 16-81 in the Gynaecological Outpatient Clinic of the Hospital de Santo Espirito de Angra do Heroismo (HSEAH). HPV DNA was amplified by Polymerase Chain Reaction using the general consensus primers PGMYO9/PGMY11. Commercially available Papillomavirus Clinical Arrays kits (Genomica) were used to perform HPV genotyping. 30 women were HPV positive, with a median age of 41 years old. Our results show that the overall HPV prevalence was 10.49%. Seventeen genotypes were identified, including 58.82% high risk, 17.65% low risk and 23.53% undetermined risk. CONCLUSION: Unlike other epidemiological studies, HPV31 was the most frequent type (26.67%) in Terceira Island, followed by HPV16 (10.00%), HPV51, HPV53, HPV70 and HPV82 (6.67%). Further studies are needed to investigate if the HPV types found in our population are associated with the risk of progression to high-grade squamous intraepithelial lesions or cervical cancer. PMID- 18426590 TI - An interdisciplinary clinical practice model for the management of low-back pain in primary care: the CLIP project. AB - BACKGROUND: Low-back pain is responsible for significant disability and costs in industrialized countries. Only a minority of subjects suffering from low-back pain will develop persistent disability. However, this minority is responsible for the majority of costs and has the poorest health outcomes. The objective of the Clinic on Low-back pain in Interdisciplinary Practice (CLIP) project was to develop a primary care interdisciplinary practice model for the clinical management of low-back pain and the prevention of persistent disability. METHODS: Using previously published guidelines, systematic reviews and meta-analyses, a clinical management model for low-back pain was developed by the project team. A structured process facilitating discussions on this model among researchers, stakeholders and clinicians was created. The model was revised following these exchanges, without deviating from the evidence. RESULTS: A model consisting of nine elements on clinical management of low-back pain and prevention of persistent disability was developed. The model's two core elements for the prevention of persistent disability are the following: 1) the evaluation of the prognosis at the fourth week of disability, and of key modifiable barriers to return to usual activities if the prognosis is unfavourable; 2) the evaluation of the patient's perceived disability every four weeks, with the evaluation and management of barriers to return to usual activities if perceived disability has not sufficiently improved. CONCLUSION: A primary care interdisciplinary model aimed at improving quality and continuity of care for patients with low-back pain was developed. The effectiveness, efficiency and applicability of the CLIP model in preventing persistent disability in patients suffering from low-back pain should be assessed. PMID- 18426591 TI - Cutpoints for mild, moderate and severe pain in patients with osteoarthritis of the hip or knee ready for joint replacement surgery. AB - BACKGROUND: Cutpoints (CPs) for mild, moderate and severe pain are established and used primarily in cancer pain. In this study, we wanted to determine the optimal CPs for mild, moderate, and severe pain in joint replacement surgery candidates with osteoarthritis (OA) of the hip or knee, and to validate the different CPs. METHODS: Patients (n = 353) completed the Brief Pain Inventory (BPI), the WOMAC Arthritis Index, and the SF-36 health status measure. Optimal CPs for categorizing average pain with three severity levels were derived using multivariate analysis of variance, using different CP sets for average pain as the independent variable and seven interference items from the BPI as the dependent variable. To validate the CPs, we assessed if patients in the three pain severity groups differed in pain as assessed with WOMAC and SF-36, and if BPI average pain with the optimal CPs resulted in higher correlation with pain dimensions of the WOMAC and SF-36 than other CPs. RESULTS: The optimal CPs on the 0-10 point BPI scale were CP (4,6) among hip patients and CP (4,7) among knee patients. The resulting pain severity groups differed in pain, as assessed with other scales than those used to derive the CPs. The optimal CPs had the highest association of average pain with WOMAC pain scores. CONCLUSION: CPs for pain severity differed somewhat for patients with OA of the hip and knee. The association of BPI average pain scores categorized according to the optimal CPs with WOMAC pain scores supports the validity of the derived optimal CPs. PMID- 18426592 TI - Nav1.7 expression is increased in painful human dental pulp. AB - BACKGROUND: Animal studies and a few human studies have shown a change in sodium channel (NaCh) expression after inflammatory lesions, and this change is implicated in the generation of pain states. We are using the extracted human tooth as a model system to study peripheral pain mechanisms and here examine the expression of the Nav1.7 NaCh isoform in normal and painful samples. Pulpal sections were labeled with antibodies against: 1) Nav1.7, N52 and PGP9.5, and 2) Nav1.7, caspr (a paranodal protein used to identify nodes of Ranvier), and myelin basic protein (MBP), and a z-series of optically-sectioned images were obtained with the confocal microscope. Nav1.7-immunofluorescence was quantified in N52/PGP9.5-identified nerve fibers with NIH ImageJ software, while Nav1.7 expression in myelinated fibers at caspr-identified nodal sites was evaluated and further characterized as either typical or atypical as based on caspr relationships. RESULTS: Results show a significant increase in nerve area with Nav1.7 expression within coronal and radicular fiber bundles and increased expression at typical and atypical caspr-identified nodal sites in painful samples. Painful samples also showed an augmentation of Nav1.7 within localized areas that lacked MBP, including those associated with atypical caspr-identified sites, thus identifying NaCh remodeling within demyelinating axons as the basis for a possible pulpal pain mechanism. CONCLUSION: This study identifies the increased axonal expression and augmentation of Nav1.7 at intact and remodeling/demyelinating nodes within the painful human dental pulp where these changes may contribute to constant, increased evoked and spontaneous pain responses that characterize the pain associated with toothache. PMID- 18426593 TI - Linking Cytoscape and the corynebacterial reference database CoryneRegNet. AB - BACKGROUND: Recently, the research community has seen an influx of data relating to transcriptional regulatory interactions of Corynebacteria, organisms that are highly relevant to fields of systems biology, biotechnology, and human medicine. Information derived from DNA microarray experiments, computational predictions, and literature has opened the way for the graph-based analysis, visualization, and reconstruction of transcriptional regulatory networks across entire organisms. The reference database for corynebacterial gene regulatory networks CoryneRegNet provides methods for data storage and data exchange in a well structured manner. Additional information on the model organism Escherichia coli K12 obtained from RegulonDB has been integrated. Generally, gene regulatory networks can be visualized as graphs by drawing directed edges between nodes, where a node represents a gene and an edge corresponds to a typed regulatory interaction. Cytoscape is an open-source software project whose aim is to provide graph-based visualization and analysis for biological networks. Its architecture allows the development and integration of user-made plugins to enhance core functionalities. RESULTS: We introduce two novel plugins for the Cytoscape environment designed to enhance in silico studies of procaryotic transcriptional regulatory networks. Our plugins leverage the information from the cornyebacterial reference database CoryneRegNet with the graph analysis capabilities of Cytoscape. CoryneRegNetLoader queries the CoryneRegNet database to extract a gene regulatory network represented as a directed graph. Additional information is stored as node/edge attributes within the graph. COMA facilitates consistency checks for gene expression studies given a gene regulatory network. COMA tests whether all gene expression levels correlate properly with the given network topology. CONCLUSION: The plugins facilitate in silico studies of procaryotic transcriptional gene regulation, particularly in Corynebacteria and E. coli, by combining the knowledge from the corynebacterial reference database with the graph analysis capabilities of Cytoscape, which is one of the most widely used tools for biological network analyses. PMID- 18426594 TI - Investigation of a novel approach to scoring Giemsa-stained malaria-infected thin blood films. AB - Daily assessment of the percentage of erythrocytes that are infected ('percent parasitaemia') across a time-course is a necessary step in many experimental studies of malaria, but represents a time-consuming and unpopular task among researchers. The most common method is extensive microscopic examination of Giemsa-stained thin blood-films. This study explored a method for the assessment of percent-parasitaemia that does not require extended periods of microscopy and results in a descriptive and permanent record of parasitaemia data that is highly amenable to subsequent 'data-mining'. Digital photography was utilized in conjunction with a basic purpose-written computer programme to test the viability of the concept. Partial automation of the determination of percent parasitaemia was then explored, resulting in the successful customization of commercially available broad-spectrum image analysis software towards this aim. Lastly, automated discrimination between infected and uninfected RBCs based on analysis of digital parameters of individual cell images was explored in an effort to completely automate the calculation of an accurate percent-parasitaemia. PMID- 18426595 TI - Reduction in magnetic resonance imaging T2 burden of disease in patients with relapsing-remitting multiple sclerosis: analysis of 48-week data from the EVIDENCE (EVidence of Interferon Dose-response: European North American Comparative Efficacy) study. AB - BACKGROUND: The EVIDENCE (EVidence of Interferon Dose-response: European North American Comparative Efficacy) study was an international, randomized, open label, assessor-blinded, parallel-group study assessing the efficacy and tolerability of interferon (IFN) beta-1a, 44 mcg subcutaneously (sc) three times weekly (tiw), and IFN beta-1a, 30 mcg intramuscularly (im) once weekly (qw), in patients with relapsing-remitting multiple sclerosis (RRMS). The aim of this analysis was to assess whether reductions in T2 burden of disease (BOD) were greater for patients receiving IFN beta-1a, 44 mcg sc tiw, than for those treated with IFN beta-1a, 30 mcg im qw, and to assess the impact of neutralizing antibodies (NAbs). METHODS: A post-hoc analysis was performed on magnetic resonance imaging (MRI) data collected prospectively from the EVIDENCE study. The analysis included all patients with evaluable T2 MRI scans at the start of dosing and at week 48, and those who received at least one drug dose (n = 553). Lesions were identified by a radiologist blinded to treatment codes and the total volume of T2 lesions (BOD) was reported in mm3. RESULTS: Both median percentage decreases and absolute reduction in BOD were greater in the IFN beta-1a, 44 mcg sc tiw, treatment group. The adjusted mean treatment difference in percentage change in BOD from baseline to week 48 showed a significant treatment benefit for patients treated with IFN beta-1a, 44 mcg sc tiw, over those treated with IFN beta-1a, 30 mcg im qw (-4.6%; standard error: 2.6%; p = 0.002). The presence of NAbs reduced the effect of IFN beta-1a 44, mcg sc tiw, on BOD, but BOD changes were still similar to those seen with IFN beta-1a, 30 mcg im qw. CONCLUSION: Patients with RRMS treated with IFN beta-1a, 44 mcg sc tiw, had greater reduction in T2 BOD after 48 weeks than those treated with IFN beta-1a, 30 mcg im qw, which is consistent with other clinical and MRI outcome measures in the EVIDENCE study. In patients testing positive for NAbs (NAb+) to IFN beta-1a 44 mcg sc tiw, changes in BOD were smaller than in NAb negative (NAb-) patients, but similar to those receiving IFN beta-1a, 30 mcg im qw. PMID- 18426596 TI - Diagnosis of ventilator-associated pneumonia: a systematic review of the literature. AB - INTRODUCTION: Early, accurate diagnosis is fundamental in the management of patients with ventilator-associated pneumonia (VAP). The aim of this qualitative review was to compare various criteria of diagnosing VAP in the intensive care unit (ICU) with a special emphasis on the value of clinical diagnosis, microbiological culture techniques, and biomarkers of host response. METHODS: A MEDLINE search was performed using the keyword 'ventilator associated pneumonia' AND 'diagnosis'. Our search was limited to human studies published between January 1966 and June 2007. Only studies of at least 25 adult patients were included. Predefined variables were collected, including year of publication, study design (prospective/retrospective), number of patients included, and disease group. RESULTS: Of 572 articles fulfilling the initial search criteria, 159 articles were chosen for detailed review of the full text. A total of 64 articles fulfilled the inclusion criteria and were included in our review. Clinical criteria, used in combination, may be helpful in diagnosing VAP, however, the considerable inter-observer variability and the moderate performance should be taken in account. Bacteriologic data do not increase the accuracy of diagnosis as compared to clinical diagnosis. Quantitative cultures obtained by different methods seem to be rather equivalent in diagnosing VAP. Blood cultures are relatively insensitive to diagnose pneumonia. The rapid availability of cytological data, including inflammatory cells and Gram stains, may be useful in initial therapeutic decisions in patients with suspected VAP. C-reactive protein, procalcitonin, and soluble triggering receptor expressed on myeloid cells are promising biomarkers in diagnosing VAP. CONCLUSION: An integrated approach should be followed in diagnosing and treating patients with VAP, including early antibiotic therapy and subsequent rectification according to clinical response and results of bacteriologic cultures. PMID- 18426597 TI - Development and validation of an Eating Disorders Symptom Impact Scale (EDSIS) for carers of people with eating disorders. AB - BACKGROUND: Family members of relatives with eating disorders experience high levels of distress due to the difficulties in their care giving role. However no measures have been developed to measure the specific impact that an individual with an eating disorder has on family life. The aim of this study was to develop a measure to assess the specific caregiving burden of both anorexia nervosa and bulimia nervosa. A secondary aim was to examine whether this measure was sensitive to change. METHODS: A new scale, the Eating Disorders Symptom Impact Scale (EDSIS), was generated by a panel of clinicians and researchers based upon quantitative and qualitative work with carers and reviewed by a panel of "expert carers". A cross-sectional study was conducted among carers of relatives with an eating disorder to examine the properties of the new scale. In addition, participants from an ongoing pre-and-post design study completed several self report questionnaires to assess the sensitivity of the EDSIS to change. RESULTS: A sample of 196 carers of relatives with an eating disorder aged 25-68 compted the scale. A 24-item EDSIS scale was derived with four factors: nutrition, guilt, dysregulated behaviour and social isolation. These explained 58.4% of the variance in carer distress. Reliability was acceptable (Cronbach's alpha ranged from 0.84 to 0.90). The convergent validity of the EDSIS subscales was moderately supported by correlations with a general caregiving measure (Experience of Caregiving Inventory (ECI), r = 0.42 to 0.60), psychological distress (General Health Questionnaire (GHQ-12), r = 0.33) and perceived functioning of the relative (Children Global Assessment Scale (CGAS), r = -30). A sample of 57 primary caregivers completed pre-post intervention assessments and the overall scale (t = 2.3, p < 0.05) and guilt subscale (t = 3.4, p < 0.01) were sensitive to change following a skills training workshop. CONCLUSION: The EDSIS instrument has good psychometric properties and may be of value to assess the impact of eating disorder symptoms on family members. It may be of value to highlight carers' needs and to monitor the effectiveness of family based interventions. PMID- 18426598 TI - Intrahepatic cholestasis in subclinical and overt hyperthyroidism: two case reports. AB - INTRODUCTION: Non-specific abnormalities in liver function tests might accompany the clinical course of hyperthyroidism. Hyperthyroidism can cause the elevation of hepatic enzymes and bilirubin. Jaundice is rare in overt hyperthyroidism, especially in subclinical hyperthyroidism. On the other hand, the use of anti thyroid drugs has rarely been associated with toxic hepatitis and cholestatic jaundice. CASE PRESENTATION: Here we present two cases of cholestasis that accompanied two distinct forms of clinical hyperthyroidism. The first patient had a clinical presentation of severe cholestasis in the absence of congestive failure related to hyperthyroidism. The second case had developed intrahepatic cholestasis in the presence of subclinical hyperthyroidism, and improved with rifampicin treatment. CONCLUSION: Hyperthyroidism should be a consideration in non-specific liver dysfunction. PMID- 18426599 TI - Sex and the city: differences in disease- and disability-free life years, and active community participation of elderly men and women in 7 cities in Latin America and the Caribbean. AB - BACKGROUND: The world's population is ageing, and four of the top 10 most rapidly ageing developing nations are from the region of Latin America and the Caribbean (LAC). Although an ageing population heralds likely increases in chronic disease, disability-related dependence, and economic burden, the societal contribution of the chronically ill or those with disability is not often measured. METHODS: We calculated country-specific prevalences of 'disability' (difficulty with at least one activity of daily living), 'disease' and 'co-morbidity' (presence of at least one, and at least two, of seven chronic diseases/conditions, respectively), and 'active community engagement' (using five levels of community participation, from less than weekly community contact to voluntary or paid work) in seven LAC cities. We estimated remaining life expectancy (LE) with and without disability, disease and co-morbidity, and investigated age, sex, and regional variations in disability-free LE. Finally, we modeled the association of disease, co-morbidity and disability with active community participation using an ordinal regression model, adjusted for depression. RESULTS: Overall, 77% of the LAC elderly had at least one chronic disease/condition, 44% had co-morbidity and 19% had a disability. The proportion of disability-free LE declined between the youngest (60-64 years) and the eldest (90 years and over) age-groups for both men (from 85% to 55%) and women (from 75% to 45%). Disease-free and co-morbidity-free LE, however, remained at approximately 30% and 62%, respectively, for men (20% and 48% for women), until 80-84 years of age, then increased. Only Bridgetown's participants had statistically significantly longer disability-free LE than the regional average (IRR = 1.08; 95%CI 1.05-1.10; p < 0.001). Only Santiago's participants had disability-free LE which was shorter than the regional average (IRR = 0.94; 95%CI 0.92-0.97; p < 0.001). There was 75% active community participation overall, with more women than men involved in active help (49% vs 32%, respectively) and more men involved in voluntary/paid work (46% vs 25%, respectively). There was either no, or borderline significance in the association between having one or more diseases/conditions and active community engagement for both sexes. These associations were limited by depression (odds ratio [OR] reduced by 15-17% for men, and by 8-11% for women), and only remained statistically significant in men. However, disability remained statistically significantly associated with less community engagement after adjusting for depression (OR = 0.58, 95%CI 0.49-0.69, p < 0.001 for women and OR = 0.50, 95%CI 0.47-0.65, p < 0.001 for men). CONCLUSION: There is an increasing burden of disease and disability with older age across the LAC region. As these nations cope with resulting social and economic demands, governments and civic societies must continue to develop and maintain opportunities for community participation by this increasingly frail, but actively engaged group. PMID- 18426600 TI - A middle-aged female with recurrent sinopulmonary infections: a case report. AB - INTRODUCTION: Common variable immunodeficiency (CVID) is a form of severe antibody deficiency with an estimated prevalence of 1 in 25,000 to 1 in 100,000. The disorder apparently results from currently undefined immune deregulations resulting in failed B-cell differentiation with impaired secretion of immunoglobulins. It has a broad range of clinical symptoms including recurrent infections of the respiratory tract, chronic lung disease, autoimmune diseases, liver and gastrointestinal disorders, granulomatous infiltrations, lymphoma and solid tumors. CASE PRESENTATION: A 42-year-old Caucasian female presented with a one-day history of high-grade fever and productive cough associated with retrosternal chest pain. The patient had been discharged one week prior after a prolonged stay in an intensive care unit with multiorgan failure requiring temporary hemodialysis for two weeks secondary to sepsis. Past medical history was significant for chronic obstructive pulmonary disease, recurrent pneumonias and recurrent sinus infections since adolescence. She had a temperature of 99.8 degrees F, was tachycardic (137/min), tachypneic (26/min) with a blood pressure of 109/59 mmHg and oxygen saturation of 88% on 2 l/min nasal oxygen. Physical examination was significant for bibasilar rhonchi. Laboratory data were significant for leukocytosis of 15,700/mm3. Chest X-ray demonstrated bibasilar infiltrates. The patient was started on intravenous levofloxacin and vancomycin, and sputum gram stain and cultures were performed. Given the patient's recurrent respiratory infections, an underlying immunologic disorder was considered. Work up revealed immunoglobulin A (IgA) 11 mg/dl (normal 70-400 mg/dl), immunoglobulin M (IgM) 2 mg/dl (normal 40-230 mg/dl) and IgG 53 mg/dl (normal 700-1,600 mg/dl). The patient was diagnosed with CVID and started on intravenous immunoglobulin. She was initially started on a four-week regimen of intravenous immunoglobulin, which was later switched to a three-week regimen as the patient had respiratory infections on the four-week regimen. She remained asymptomatic on a three times/week intravenous immunoglobulin regimen. CONCLUSION: This case emphasizes the need for a high index of clinical suspicion for CVID in patients presenting with recurrent sinopulmonary infections. Although intravenous immunoglobulin provides improvement in these patients, early diagnosis is the key to preventing significant morbidity and mortality and improving prognosis. PMID- 18426602 TI - Development and validation of the Measure of Indigenous Racism Experiences (MIRE). AB - BACKGROUND: In recent decades there has been increasing evidence of a relationship between self-reported racism and health. Although a plethora of instruments to measure racism have been developed, very few have been described conceptually or psychometrically Furthermore, this research field has been limited by a dearth of instruments that examine reactions/responses to racism and by a restricted focus on African American populations. METHODS: In response to these limitations, the 31-item Measure of Indigenous Racism Experiences (MIRE) was developed to assess self-reported racism for Indigenous Australians. This paper describes the development of the MIRE together with an opportunistic examination of its content, construct and convergent validity in a population health study involving 312 Indigenous Australians. RESULTS: Focus group research supported the content validity of the MIRE, and inter-item/scale correlations suggested good construct validity. A good fit with a priori conceptual dimensions was demonstrated in factor analysis, and convergence with a separate item on discrimination was satisfactory. CONCLUSION: The MIRE has considerable utility as an instrument that can assess multiple facets of racism together with responses/reactions to racism among indigenous populations and, potentially, among other ethnic/racial groups. PMID- 18426601 TI - The body politic: the relationship between stigma and obesity-associated disease. AB - BACKGROUND: It is commonly believed that the pathophysiology of obesity arises from adiposity. In this paper, I forward a complementary explanation; this pathophysiology arises not from adiposity alone, but also from the psychological stress induced by the social stigma associated with being obese. METHODS: In this study, I pursue novel lines of evidence to explore the possibility that obesity associated stigma produces obesity-associated medical conditions. I also entertain alternative hypotheses that might explain the observed relationships. RESULTS: I forward four lines of evidence supporting the hypothesis that psychological stress plays a role in the adiposity-health association. First, body mass index (BMI) is a strong predictor of serological biomarkers of stress. Second, obesity and stress are linked to the same diseases. Third, body norms appear to be strong determinants of morbidity and mortality among obese persons; obese whites and women - the two groups most affected by weight-related stigma in surveys - disproportionately suffer from excess mortality. Finally, statistical models suggest that the desire to lose weight is an important driver of weight related morbidity when BMI is held constant. CONCLUSION: Obese persons experience a high degree of stress, and this stress plausibly explains a portion of the BMI health association. Thus, the obesity epidemic may, in part, be driven by social constructs surrounding body image norms. PMID- 18426603 TI - Synergistic effects of high fat feeding and apolipoprotein E deletion on enterocytic amyloid-beta abundance. AB - BACKGROUND: Amyloid-beta (Abeta), a key protein found in amyloid plaques of subjects with Alzheimer's disease is expressed in the absorptive epithelial cells of the small intestine. Ingestion of saturated fat significantly enhances enterocytic Abeta abundance whereas fasting abolishes expression. Apolipoprotein (apo) E has been shown to directly modulate Abeta biogenesis in liver and neuronal cells but it's effect in enterocytes is not known. In addition, apo E modulates villi length, which may indirectly modulate Abeta as a consequence of differences in lipid absorption. This study compared Abeta abundance and villi length in wild-type (WT) and apo E knockout (KO) mice maintained on either a low fat or high-fat diet. Wild-type C57BL/6J and apo E KO mice were randomised for six-months to a diet containing either 4% (w/w) unsaturated fats, or chow comprising 16% saturated fats and 1% cholesterol. Quantitative immunohistochemistry was used to assess Abeta abundance in small intestinal enterocytes. Apo E KO mice given the low-fat diet had similar enterocytic Abeta abundance compared to WT controls. RESULTS: The saturated fat diet substantially increased enterocytic Abeta in WT and in apo E KO mice, however the effect was greater in the latter. Villi height was significantly greater in apo E KO mice than for WT controls when given the low-fat diet. However, WT mice had comparable villi length to apo E KO when fed the saturated fat and cholesterol enriched diet. There was no effect of the high-fat diet on villi length in apo E KO mice. CONCLUSION: The findings of this study are consistent with the notion that lipid substrate availability modulates enterocytic Abeta. Apo E may influence enterocytic lipid availability by modulating absorptive capacity. PMID- 18426604 TI - DNA-dependent protein kinase catalytic subunit modulates the stability of c-Myc oncoprotein. AB - BACKGROUND: C-Myc is a short-lived oncoprotein that is destroyed by ubiquitin mediated proteolysis. Dysregulated accumulation of c-Myc commonly occurs in human cancers. Some of those cases with the dysregulated c-Myc protein accumulation are attributed to gene amplification or increased mRNA expression. However, the abnormal accumulation of c-Myc protein is also a common finding in human cancers with normal copy number and transcription level of c-Myc gene. It seems that the mechanistic dysregulation in the control of c-Myc protein stabilization is another important hallmark associated with c-Myc accumulation in cancer cells. Here we report a novel mechanistic pathway through which DNA-dependent protein kinase catalytic subunit (DNA-PKcs) modulates the stability of c-Myc protein. RESULTS: Firstly, siRNA-mediated silencing of DNA-PKcs strikingly downregulated c Myc protein levels in HeLa and HepG2 cells, and simultaneously decreased cell proliferation. The c-Myc protein level in DNA-PKcs deficient human glioma M059J cells was also found much lower than that in DNA-PKcs efficient M059K cells. ATM deficiency does not affect c-Myc expression level. Silencing of DNA-PKcs in HeLa cells resulted in a decreased stability of c-Myc protein, which was associated the increasing of c-Myc phosphorylation on Thr58/Ser62 and ubiquitination level. Phosphorylation of Akt on Ser473, a substrate of DNA-PKcs was found decreased in DNA-PKcs deficient cells. As the consequence, the phosphorylation of GSK3 beta on Ser9, a negatively regulated target of Akt, was also decreased, and which led to activation of GSK 3beta and in turn phosphorylation of c-Myc on Thr58. Moreover, inhibition of GSK3 activity by LiCl or specific siRNA molecules rescued the downregulation of c-Myc mediated by silencing DNA-PKcs. Consistent with this depressed DNA-PKcs cell model, overexpressing DNA-PKcs in normal human liver L02 cells, by sub-chronically exposing to very low dose of carcinogen 2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD), increased c-Myc protein level, the phosphorylation of Akt and GSK3 beta, as well as cell proliferation. siRNA mediated silencing of DNA-PKcs in this cell model reversed above alterations to the original levels of L02 cells. CONCLUSION: A suitable DNA-PKcs level in cells is necessary for maintaining genomic stability, while abnormal overexpression of DNA-PKcs may contribute to cell proliferation and even oncogenic transformation by stabilizing the c-Myc oncoprotein via at least the Akt/GSK3 pathway. Our results suggest DNA-PKcs a novel biological role beyond its DNA repair function. PMID- 18426605 TI - Characteristic expression of HTLV-1 basic zipper factor (HBZ) transcripts in HTLV 1 provirus-positive cells. AB - BACKGROUND: HTLV-1 causes adult T-cell leukemia (ATL). Although there have been many studies on the oncogenesis of the viral protein Tax, the precise oncogenic mechanism remains to be elucidated. Recently, a new viral factor, HTLV-1 basic Zip factor (HBZ), encoded from the minus strand mRNA was discovered and the current models of Tax-centered ATL cell pathogenesis are in conflict with this discovery. HBZs consisting of non-spliced and spliced isoforms (HBZ-SI) are thought to be implicated in viral replication and T-cell proliferation but there is little evidence on the HBZ expression profile on a large scale. RESULTS: To investigate the role of HBZ-SI in HTLV-1 provirus-positive cells, the HBZ-SI and Tax mRNA loads in samples with a mixture of infected and non-infected cells were measured and then adjusted by dividing by the HTLV-I proviral load. We show here that the HBZ-SI mRNA level is 4-fold higher than non-spliced HBZ and is expressed by almost all cells harboring HTLV-1 provirus with variable intensity. The proviral-adjusted HBZ-SI and Tax quantification revealed a characteristic imbalanced expression feature of high HBZ and low Tax expression levels in primary ATL cells or high HBZ and very high Tax levels in HTLV-1-related cell lines (cell lines) compared with a standard expression profile of low HBZ and low Tax in infected cells. Interestingly, according to the mutual Tax and HBZ expression status, HTLV-1-related cell lines were subcategorized into two groups, an ATL cell type with high HBZ and low Tax levels and another type with high Tax and either high or low HBZ, which was closely related to its cell origin. CONCLUSION: This is the first comprehensive study to evaluate the mutual expression profile of HBZ and Tax in provirus-positive cells, revealing that there are quantitative and relative characteristic features among infected cells, primary ATL cells, and cell lines. PMID- 18426606 TI - Role of Plasmodium falciparum thrombospondin-related anonymous protein in host cell interactions. AB - BACKGROUND: Thrombospondin-related anonymous protein (TRAP) is essential for sporozoite motility and for liver cell invasion. TRAP is a type 1 membrane protein that possesses multiple adhesive domains in its extracellular region. METHODS: Plasmodium falciparum TRAP (PfTRAP) and its subdomains were expressed in a mammalian expression system, and eleven different mutants generated to study interaction of PfTRAP with liver cells. Binding studies between HepG2 cell extracts and PfTRAP were performed using co-immunoprecipitation protocols. RESULTS: Five different amino acid residues of PfTRAP that are involved in liver cell binding have been identified. These PfTRAP mutants bound to heparin like the wild type PfTRAP thereby suggesting a non-heparin mediated binding of PfTRAP to liver cells. Three Src family proteins -Lyn, Lck and CrkL which interact with PfTRAP are also identified. Liver cell extracts and immunoprecipitated Src family kinases phosphorylated PfTRAP at multiple sites. An analysis of multiple TRAP sequences revealed Src homology 3 domain (SH3) binding motifs. CONCLUSION: Binding of PfTRAP to SH3-domain containing proteins like Src-family kinases and their ability to phosphorylate PfTRAP suggests a novel role for PfTRAP in cell signaling during sporozoite invasion and homing inside the liver cells. These data shed new light on TRAP-liver cell interactions. PMID- 18426607 TI - Conventionally-fractionated image-guided intensity modulated radiotherapy (IG IMRT): a safe and effective treatment for cancer spinal metastasis. AB - BACKGROUND: Treatments for cancer spinal metastasis were always palliative. This study was conducted to investigate the safety and effectiveness of IG-IMRT for these patients. METHODS: 10 metastatic lesions were treated with conventionally fractionated IG-IMRT. Daily kilovoltage cone-beam computed tomography (kV-CBCT) scan was applied to ensure accurate positioning. Plans were evaluated by the dose volume histogram (DVH) analysis. RESULTS: Before set-up correction, the positioning errors in the left-right (LR), superior-inferior (SI) and anterior posterior (AP) axes were 0.3 +/- 3.2, 0.4 +/- 4.5 and -0.2 +/- 3.9 mm, respectively. After repositioning, those errors were 0.1 +/- 0.7, 0 +/- 0.8 and 0 +/- 0.7 mm, respectively. The systematic/random uncertainties ranged 1.4-2.3/3.0 4.1 before and 0.1-0.2/0.7-0.8 mm after online set-up correction. In the original IMRT plans, the average dose of the planning target volume (PTV) was 61.9 Gy, with the spinal cord dose less than 49 Gy. Compared to the simulated PTVs based on the pre-correction CBCT, the average volume reduction of PTVs was 42.3% after online correction. Also, organ at risk (OAR) all benefited from CBCT-based set-up correction and had significant dose reduction with IGRT technique. Clinically, most patients had prompt pain relief within one month of treatment. There was no radiation-induced toxicity detected clinically during a median follow-up of 15.6 months. CONCLUSION: IG-IMRT provides a new approach to treat cancer spinal metastasis. The precise positioning ensures the implementation of optimal IMRT plan, satisfying both the dose escalation of tumor targets and the radiation tolerance of spinal cord. It might benefit the cancer patient with spinal metastasis. PMID- 18426608 TI - Length of stay and associated costs of obesity related hospital admissions in Ireland. AB - BACKGROUND: Obesity is the cause of other chronic diseases, psychological problems, obesity shortens the lifespan and puts strain on health systems. The risk associated with childhood obesity in particular, which will accelerate the development of adult morbidity and mortality, has been identified as an emerging public health problem. METHODS: To estimate the length of stay and associated hospital costs for obesity related illnesses a cost of illness study was set up. All discharges from all acute hospitals in the Republic of Ireland from 1997 to 2004 with a principal or secondary diagnostic code for obesity for all children from 6 to 18 years of age and for adults were collected.A discharge frequency was calculated by dividing obesity related discharges by the total number of diagnoses (principal and secondary) for each year. The hospital costs related to obesity was calculated based on the total number of days care. RESULTS: The discharge frequency of obesity related conditions increased from 1.14 in 1997 to 1.49 in 2004 for adults and from 0.81 to 1.37 for children. The relative length of stay (number of days in care for obesity related conditions per 1000 days of hospital care given) increased from 1.47 in 1997 to 4.16 in 2004 for children and from 3.68 in 1997 to 6.74 in 2004 for adults. Based on the 2001 figures for cost per inpatient bed day, the annual hospital cost was calculated to be 4.4 Euromillion in 1997, increasing to 13.3 Euromillion in 2004. At a 20% variable hospital cost the cost ranges from 0.9 Euromillion in 1997 to 2.7 Euromillion in 2004; a 200% increase. CONCLUSION: The annual increase in the proportion of hospital discharges related to obesity is alarming. This increase is related to a significant increase in economic costs. This paper emphasises the need for action at an early stage of life. Health promotion and primary prevention of obesity should be high on the political agenda. PMID- 18426610 TI - Trends in asthma prevalence, admission rates, and asthma deaths. AB - There is now clear evidence that asthma prevalence increased significantly, especially in developed countries, during the second part of the 20th century. What caused this increase is currently unknown. Recent reports from the United States and the United Kingdom suggest that asthma prevalence may have plateaued between 1995 and the first few years of the present century. This stabilization, and even some decrease in asthma prevalence, especially in countries with high baseline rates, was confirmed by the International Study of Asthma and Allergies in Children. The hospitalization rate for asthma (as a proportion of asthma patients) decreased significantly in the United States between 1980 and 1995, then remained stable between 2001 and 2004. However, the asthma death rate (as a proportion of subjects with asthma) did not decrease significantly during either of those periods. A better understanding of what determines the stable asthma death rate is urgently needed, especially since inhaled corticosteroids have been shown to prevent asthma deaths in persons who take them regularly. PMID- 18426611 TI - Clinical asthma syndromes and important asthma mimics. AB - Asthma is a heterogeneous disorder with multiple clinical phenotypes. Phenotypes can be grouped into clinical or physiological, trigger-defined, and inflammatory phenotypes. Treatment based on inflammatory phenotyping improves clinical measures of asthma morbidity. Further study of individual asthma phenotypes will improve understanding of their immunologic and pathologic characteristics and improve diagnosis and therapy. Because asthma is a common disorder with nonspecific presenting features, other disorders are often misdiagnosed as asthma. A high index of suspicion for alternative diagnoses must be maintained when evaluating a patient who presents with clinical features suggestive of asthma, particularly if the patient presents with atypical symptoms or fails to respond to therapy. PMID- 18426612 TI - Making the diagnosis of asthma. AB - Diagnostic tests can only increase or decrease the probability of the asthma diagnosis, so a thorough history is very important. In patients with asthma-like symptoms, spirometric evidence of airway obstruction plus a large bronchodilator response makes asthma much more likely. However, normal spirometry is common in patients with mild asthma who are not symptomatic at the time of testing, and patients with poorly controlled asthma may lack substantial bronchodilator response. Inhalation challenge test often helps confirm asthma in patients with normal spirometry. Adult smokers with intermittent respiratory symptoms may have either asthma or chronic obstructive pulmonary disease (COPD). Normal post bronchodilator spirometry rules out COPD. In patients with airway obstruction, a low diffusing capacity of the lung for carbon monoxide increases the probability of COPD and makes asthma much less likely. A high exhaled nitric oxide level makes allergic asthma more likely. Response to inhaled corticosteroids makes asthma more likely and COPD less likely. PMID- 18426613 TI - Traditional and new approaches to asthma monitoring. AB - Once the diagnosis of asthma is established, monitoring must be implemented to achieve asthma control. Because of the variability of asthma, monitoring is a long-term commitment to effectively adjust treatment and assure that therapy goals are met. This paper reviews the definition of asthma control, including the dimensions of impairment and risk, and the 2007 National Asthma Education and Prevention Program's Expert Panel Report 3, Guidelines for the Diagnosis and Management of Asthma, recommendations for periodic assessment and monitoring of effective control. New approaches to asthma monitoring, such as airway hyperresponsiveness, sputum eosinophils, exhaled nitric oxide, and pharmacogenetic measurements, will be critiqued. PMID- 18426615 TI - Safety and efficacy of beta agonists. AB - Both short-acting and long-acting beta agonists have been used for many years for the treatment of asthma. Short-acting beta agonists are life-saving and their role as rescue agents is unquestioned, but regular use is not recommended because of safety concerns and the effectiveness of asthma-controller medications. Long acting beta agonists are effective controller medications but have safety issues, so their use should be restricted to patients who are not optimally controlled on first-line controllers such as inhaled corticosteroids. The effect of the beta receptor genotype on beta agonist response is unclear but could hold promise for proper patient management. PMID- 18426616 TI - Inhaled corticosteroids in asthma management. AB - Asthma is a disorder of the lower airways, characterized by bronchial hyperresponsiveness and airflow limitation, the pathogenesis of which is yet to be fully understood. Regardless of its triggers, asthma's hallmark is a state of inflammation that, when uncontrolled, results in persistence of symptoms. Inhaled corticosteroids are established as the mainstay of asthma therapy. This paper examines what is currently available among this class of drugs, features of the ideal inhaled corticosteroid, the delivery systems, dose-response relationships, adverse effects, combination with long-acting beta agonists, equipotent doses among the different types, and several special scenarios that involve the apparent incomplete or lack of response to treatment with inhaled corticosteroids among certain subgroups of patients, such as smokers and obese individuals, and we will discuss the scientific basis of such resistance and suggest alternative approaches to therapy. PMID- 18426614 TI - Environmental issues in managing asthma. AB - Management of asthma requires attention to environmental exposures both indoors and outdoors. Americans spend most of their time indoors, where they have a greater ability to modify their environment. The indoor environment contains both pollutants (eg, particulate matter, nitrogen dioxide, secondhand smoke, and ozone) and allergens from furred pets, dust mites, cockroaches, rodents, and molds. Indoor particulate matter consists of particles generated from indoor sources such as cooking and cleaning activities, and particles that penetrate from the outdoors. Nitrogen dioxide sources include gas stoves, furnaces, and fireplaces. Indoor particulate matter and nitrogen dioxide are linked to asthma morbidity. The indoor ozone concentration is mainly influenced by the outdoor ozone concentration. The health effects of indoor ozone exposure have not been well studied. In contrast, there is substantial evidence of detrimental health effects from secondhand smoke. Guideline recommendations are not specific for optimizing indoor air quality. The 2007 National Asthma Education and Prevention Program asthma guidelines recommend eliminating indoor smoking and improving the ventilation. Though the guidelines state that there is insufficient evidence to recommend air cleaners, air cleaners and reducing activities that generate indoor pollutants may be sound practical approaches for improving the health of individuals with asthma. The guidelines are more specific about allergen avoidance; they recommend identifying allergens to which the individual is immunoglobin E sensitized and employing a multifaceted, comprehensive strategy to reduce exposure. Outdoor air pollutants that impact asthma include particulate matter, ozone, nitrogen dioxide, and sulfur dioxide, and guidelines recommend that individuals with asthma avoid exertion outdoors when these pollutants are elevated. Outdoor allergens include tree, grass, and weed pollens, which vary in concentration by season. Recommendations to reduce exposure include staying indoors, keeping windows and doors closed, using air conditioning and perhaps high-efficiency particulate arrestor (HEPA) air filters, and thorough daily washing to remove allergens from one's person. PMID- 18426617 TI - Studies on antimalarial drug susceptibility in Colombia, in relation to Pfmdr1 and Pfcrt. AB - In Colombia, Plasmodium resistance to antimalarials such as chloroquine and antifolates is a serious problem. As a result, the national Colombian health authorities are monitoring the efficacy of alternative drugs and schemes. The study of genetic polymorphisms related with drug resistance is required in the region. In vitro responses to chloroquine, quinine, mefloquine, amodiaquine, desethylamodiaquine, artesunate and dihydroartesunate were carried out by HRP ELISA. SNP analysis in Pfcrt and Pfmdr1 genes was performed by PCR-RFLP in 77 samples from the North West region of Colombia. In vitro resistance to chloroquine was high (74%), followed by mefloquine (30%) and desethylamodiaquine (30%). A positive correlation between the IC(50) of paired drugs was also detected. The allele Pfmdr1 N86 (wild) was present in 100% of the samples and 1246Y (mutant) in 92%. However, their presence did not correlate with in vitro drug resistance. Presence of the mutations K76T and N75E in Pfcrt was confirmed in all samples. Analysis of 4 codons (72, 74, 75 and 76) in pfcrt confirmed the presence of the haplotypes CMET in 91% and SMET in 9% of the samples. PMID- 18426618 TI - Inheritance and linkage analysis of co-dominant SSR markers on the Z chromosome of the silkworm (Bombyx mori L.). AB - Microsatellites or simple sequence repeats (SSRs) are co-dominant molecular markers. When we used fluorescent SSR markers to construct a linkage map for the female heterogametic silkworm (Bombyx mori, ZW), we found that some loci did not segregate in a Mendelian ratio of 1:1 in a backcross population. These loci segregated in a 3:1 ratio of single bands compared with double bands. Further examination of band patterns indicated that three types of SSR bands were present: two homozygotes and one heterozygote. In the beginning, we considered to discard these markers. By scoring male and female F1 individuals, we confirmed that these loci were located on the Z chromosome. Using the sex-linked visible mutation sch (K05) and its wild-type (C108), we constructed an F1 male backcross (BC1M) mapping population. The combination of sch backcross and SSR data enabled us to map the SSR markers to the Z chromosome. By adjusting input parameters based on these data, we were able to use Mapmaker software to construct a linkage map. This strategy takes advantage of co-dominant markers for positional cloning of genes on the Z chromosome. We localized sch to the Z chromosome relative to six SSR markers and one PCR marker, covering a total of 76.1 cM. The sch mutation is an important sex-linked visible mutation widely used in breeding of commercial silkworms (e.g. male silkworm selection rearing). Localization of the sch gene may prove helpful in cloning the gene and developing strains for marker-assisted selection in silkworm breeding. PMID- 18426619 TI - Are autosomal sex-determining factors of the housefly (Musca domestica) spreading north? AB - Multiple sex-determining factors have been found in natural populations of the housefly, Musca domestica. Their distribution seems to follow a geographical cline. The 'standard' system, with a male-determining factor, M, located on the Y chromosome, prevails at higher latitudes and altitudes. At lower latitudes and altitudes M factors have also been found on any of the five autosomes. Such populations often also harbour a dominant autosomal factor, FD, which induces female development even in the presence of several M factors. Autosomal M factors were first observed some 50 years ago. It has been hypothesized that following their initial appearance, they are spreading northwards, replacing the standard XY system, but this has never been systematically investigated. To scrutinize this hypothesis, we here compare the current distribution of autosomal M factors in continental Europe, on a transect running from Germany to southern Italy, with the distribution reported 25 years ago. Additionally, we analysed the frequencies of the FD factor, which has not been done before for European populations. In contrast to earlier predictions, we do not find a clear change in the distribution of sex-determining factors: as 25 years ago, only the standard XY system is present in the north, while autosomal M factors and the FD factor are prevalent in Italy. We discuss possible causes for this apparently stable polymorphism. PMID- 18426620 TI - Understanding the population genetic structure of coastal species (Cakile maritima): seed dispersal and the role of sea currents in determining population structure. AB - The nature and extent of long-distance seed dispersal are currently poorly understood, largely due to the inherent difficulty in detecting such a phenomenon. Genetic methods provide one of the few general approaches that offer the potential to accurately address this issue. Phenotypic and allozymic approaches were applied to characterize inter-population seed dispersal of the sea rocket (Cakile maritima, Brassicaceae), a glabrous and succulent annual herb. Genetic variation was assessed on 360 individuals sampled from nine populations. Genetic diversity across populations was high, 37% of which was represented by Qst and 16% by Fst. When genetic distances were used to construct the UPGMA dendrogram, populations were clustered into three groups at the 90% similarity level. The pattern of clustering can be explained by examining the direction of sea currents around Tunisian coasts. We have shown in this study that C. maritima seeds can survive up to 4 months immersion in sea water and up to 1 year of floating in sea water; therefore, seed dispersal between populations is possible both in terms of seed survival and current patterns. PMID- 18426621 TI - Bayesian analysis of quantitative traits using skewed distributions. AB - Statistical models for genetic evaluation often make use of Gaussian distributions. However, some new statistical developments allow the use of an asymmetric distribution for the residuals. Within this context, we analysed three different patterns for the residual term on a data set consisting of 63 208 litter-size records, belonging to 19 255 sows, with a pedigree including 27 911 individuals. The three different residual distributions were: (1) Gaussian distribution, (2) asymmetric Gaussian distribution and (3) asymmetric Gaussian distribution with a hierarchical scheme for the asymmetry parameter. The operational model always included order of parity and herd-year-season as systematic effects, and the permanent environmental and infinitesimal genetic effect of each sow as random effects. The most suitable model using the deviance information criterion (DIC) and posterior predictive checking was model 3. This implies systematic, additive genetic and permanent environmental control of both litter size and the asymmetry parameter of the residual distribution. The asymmetry parameter can be understood as a measure of sensitivity to negative (or positive) environmental influences on phenotypes. The posterior mean (standard deviation) of the additive genetic variance was 0.28 (0.06) for litter size and 0.07 (0.01) for the asymmetry parameter. The posterior mean (standard deviation) of the additive genetic correlation between litter size and the asymmetry parameter was 0.21 (0.07). PMID- 18426622 TI - A proximal decomposition of inbreeding, coancestry and contributions. AB - Wright's (1922) coalescence probabilistic method decomposes the average coancestry or the average inbreeding of the population of interest into contributions of ancestors who are directly responsible for coancestry. These ancestors are less numerous and younger than those given by Caballero & Toro's (2000) method. Then, Wright's method yields a more parsimonious representation of the origin of coancestry or inbreeding. Results of Wright's method, called proximal because ancestors are younger, can be obtained from the latter method using a simple matrix transformation. Detailed algorithms are presented. Both methods can be used for assessing the overall efficiency of a breeding scheme in balancing genetic gain and coancestry rate, yielding very similar results when generations are discrete. PMID- 18426623 TI - Restricting coancestry and inbreeding at a specific position on the genome by using optimized selection. AB - Over recent years, selection methodologies have been developed to allow the maximization of genetic gain whilst constraining the rate of inbreeding. The desired rate of inbreeding is achieved by constraining the group coancestry using the numerator relationship matrix computed from pedigree. It is shown that when the method is applied to mixed inheritance models, where a QTL is segregating together with polygenes, the rate of inbreeding achieved in the region around a QTL is greater than the desired level. The constraint on group coancestry at specific positions around the QTL is achieved by using a relationship matrix computed from pedigree and genetic markers. However, the rate of inbreeding realized at the position of constraint is lower than that expected given the assumed relationship between group coancestry and the subsequent rate of inbreeding. The use of markers in the calculation of the relationship matrix allows the selection of candidates with very low or zero relationships because they are homozygous for alternative alleles, which results in a heterozygosity amongst their offspring higher than would be expected given their allele frequencies. A generation of random selection restored the expected relationship between group coancestry and inbreeding. PMID- 18426624 TI - Deviations from Hardy-Weinberg proportions for multiple alleles under viability selection. AB - Departures of genotype frequencies from Hardy-Weinberg proportions (HWP) for a single autosomal locus due to viability selection in a random mating population have been studied only for the two-allele case. In this article, the analysis of deviations from HWP due to constant viability selection is extended to multiple alleles. The deviations for an autosomal locus with k alleles are measured by means of k fii fixation indices for homozygotes and k(k-1)/2 fij fixation indices for heterozygotes, and expressions are obtained for these indices (FIS statistics) under the multiallele viability model. Furthermore, expressions for fii and fij when the multiallele polymorphism is at stable equilibrium are also derived and it is demonstrated that the pattern of multiallele Hardy-Weinberg deviations at equilibrium is characterized by a global heterozygote excess and a deficiency of each of the homozygotes. This pattern may be useful for detecting whether a given multiallelic polymorphism is at stable equilibrium in the population due to viability selection. An analysis of Hardy-Weinberg deviations from published data for the three-allele polymorphism at the beta-globin locus in human populations from West Africa is presented for illustration. PMID- 18426625 TI - Core curriculum in Emergency Medicine. PMID- 18426626 TI - Individual titration of propofol plasma target improves anaesthetic stability in patients undergoing major abdominal surgery: a comparison with manually controlled infusion. AB - BACKGROUND AND OBJECTIVE: The impact of anaesthesia using target-controlled infusion with propofol on intraoperative stability, recovery and cost compared to manually controlled infusion has been evaluated with inconsistent results. We studied a new device that allows more individual titration of propofol target controlled infusion by using the effect-site concentration at the loss of eyelash reflex to predict the maintenance infusion rate (FM-TCI). METHODS: Fifty-six patients undergoing major abdominal surgery lasting >2 h were randomly assigned to receive either FM-TCI (n = 28) or MCI-controlled (n = 28) anaesthesia. Both groups were Bispectral Index-monitored and thoracic epidural analgesia was established. Anaesthetic stability, incidence of haemodynamic abnormalities, time to extubation, propofol consumption and patient satisfaction were assessed. RESULTS: In the FM-TCI group, a significantly improved anaesthetic stability was achieved (0.43 +/- 0.44 vs. 1.31 +/- 0.78 adjustments of propofol infusion per patient per hour, P = 0.003) and time to extubation was significantly shorter (9.6 +/- 2.1 vs. 15.7 +/- 9.6 min P = 0.011). With FM-TCI, propofol consumption was significantly lower. Haemodynamic stability and patient satisfaction did not differ between the groups. CONCLUSION: FM-TCI helps to provide more stable anaesthesia conditions requiring less-frequent adjustments of the propofol infusion compared to manually controlled infusion in patients undergoing major abdominal surgery. PMID- 18426627 TI - Preoperatively administered ramosetron oral disintegrating tablets for preventing nausea and vomiting associated with patient-controlled analgesia in breast cancer patients. AB - BACKGROUND AND OBJECTIVES: The purpose of this study was to investigate the acceptability and therapeutic efficacy of a preoperative single administration of long-acting 5-hydroxytryptamine type 3 (5-HT 3) receptor antagonist in an orally disintegrating tablet formulation, ramosetron, in breast cancer patients. METHODS: Two hundred and forty women, ASA I-II, aged 24-60 yr, undergoing elective breast cancer surgery, were randomized. A standardized anaesthetic technique was used. Patients were assigned to receive one of three treatment regimens (n = 80 in each group): no prophylactic antiemetics (Group A), single prophylactic intravenous injection of ramosetron 0.1 mg at the completion of surgery (Group B) or preoperatively oral administration of 0.1 mg of ramosetron (Group C). Episodes of nausea and vomiting, the use of rescue antiemetic treatment, degree of pain, adverse events and level of satisfaction were recorded. RESULTS: The overall incidence of nausea and vomiting during the first 24 h after the recovery in Groups B (27.8%) and C (25%) was decreased significantly compared with Group A (75.3%). The frequency of the use of rescue antiemetics was significantly lower in Group C (5.0%) compared with Groups A (53.2%) and B (15.2%). The patients in Group C were more satisfied with control of postoperative nausea and vomiting than others. CONCLUSION: Preoperative oral administration of ramosetron at a dose of 0.1 mg is an acceptable and effective way of reducing the incidence of postoperative nausea and vomiting in breast cancer patients. PMID- 18426629 TI - Epidural volume extension and role of baricity. PMID- 18426630 TI - Overweight and health problems of the lower extremities: osteoarthritis, pain and disability. AB - AIM: To examine the association between overweight and health problems of the lower extremities, i.e. osteoarthritis (OA), pain and disability. METHODS: Cross sectional data from the Dutch population-based Musculoskeletal Conditions & Consequences Cohort (DMC3), comprising a random sample from the Dutch population aged >25 years (n 3664), were analysed using multivariate logistic regression. Overweight was defined as BMI > or = 25.0 kg/m2, moderate overweight as 25.0 kg/m2 < or = BMI < 30.0 kg/m2 and obesity as BMI > or = 30.0 kg/m2. Health problems of the lower extremities were: (i) self-reported OA of the hip or knee as told by a doctor; (ii) presence of self-reported chronic pain (>3 months) of the lower extremities; and (iii) disabilities in mobility as measured by the Euroqol questionnaire (EQ-5D). RESULTS: Moderate overweight was associated with self-reported OA of the hip or knee (OR = 1.7; 95 % CI 1.4, 2.1), chronic pain of the lower extremities at one or more location(s) (OR = 1.6; 95 % CI 1.3, 1.9) and disability in mobility (OR = 1.7; 95 % CI 1.4, 2.0). For obesity these odds were higher: 2.8 (95 % CI 2.1, 3.7), 2.5 (95 % CI 1.9, 3.2) and 3.0 (95 % CI 2.3, 3.9), respectively. Also, among those with OA, moderate overweight and obesity were associated with disability in mobility. CONCLUSION: There is a strong association between overweight/obesity and health problems of the lower extremities, i.e. OA, pain and disability. The increasing prevalence of overweight and obesity worldwide urges for public health action not only for diabetes and heart disease, but also OA. PMID- 18426631 TI - Maternal lactation characteristics after consumption of an alcoholic soup during the postpartum 'doing-the-month' ritual. AB - OBJECTIVE: The present study examined whether ethanol exposure influences lactation parameters. Specifically, selected constituents in maternal blood and milk and the lactation performance of Chinese lactating mothers were evaluated after they had consumed chicken soup flavoured with sesame oil and rice wine (CSSR), a diet traditionally prescribed during the postpartum 'doing-the-month' ritual. DESIGN: Twenty-three lactating mothers were examined. Informed consent was obtained from each subject. Each subject was tested on two occasions separated by a week. The target alcohol dosage was 0.3 g/kg body weight. Milk and blood samples were collected prior to consumption of soup and at 120 and 150 min, respectively, after consumption. Levels of various constituents were measured. The time for ejection of the first milk droplet and total milk volume yielded were also measured. RESULTS: Consumption of CSSR influenced TAG, insulin and lactate levels in maternal blood. Likewise, consumption of the soup affected milk composition and its nutritional status, particularly total protein, TAG, fatty acid, beta-hydroxybutyrate and lactate levels. CSSR intake significantly affected TAG and lactate levels in milk. The time for the first milk droplet to be ejected was significantly longer in the CSSR group, indicating that the milk-ejecting reflex is inhibited. However, blood prolactin level increased slightly after ethanol intake. Milk yields were reduced after ingestion of CSSR although the difference was not statistically significant. CONCLUSION: Consumption of CSSR affects not only the composition of maternal blood and milk, but also lactation performance. These findings suggest that an alcoholic diet should be avoided during lactation. PMID- 18426632 TI - Relationship between BMI and blood pressure in girls and boys. AB - AIM: To investigate the relationship between BMI and blood pressure as this is of crucial interest in evaluating both public health and the clinical impact of the so-called obesity epidemic. METHODS: Data were gathered from 1899 children aged between 6 and 14 years, analysing and evaluating a possible relationship between BMI and systolic and diastolic blood pressure values for both girls and boys. Each child was classified on the basis of age- and sex-specific BMI percentile as normal weight (<85th percentile), overweight (95th percentile). RESULTS: In comparisons among age BMI percentile groups, systolic and diastolic blood pressure values were higher in obese and overweight groups than in normal weight groups for both sexes. Although BMI among girls was higher than among boys in all three percentile groups, there were no significant differences between sexes with respect to blood pressure values. CONCLUSION: The present findings emphasize the importance of the prevention of obesity in order to prevent future related problems such as hypertension in children and adolescents. PMID- 18426633 TI - Predictors of usage and fatty acid composition of cooking fats in Bogota, Colombia. AB - OBJECTIVE: To examine correlates of home usage of commercially available cooking fats in Bogota, Colombia and to determine their fatty acid composition. DESIGN: Cross-sectional survey. SETTING: Bogota, Colombia. SUBJECTS: A representative sample of low- and middle-income families (n 2408). RESULTS: The types of fat primarily used for cooking at home were mixed vegetable oils (66 %), sunflower oil (21 %) and other oils/fats including margarine (13 %). In multivariate analysis, usage of sunflower oil as the primary cooking fat was positively related to home ownership, age of the father and health as a reason for choosing the main cooking fat, and inversely associated with the number of people per room and an index of household food insecurity. The trans fat content of sunflower oil was unexpectedly higher (mean 4.2 %, range 2.2-8.6) than that of the vegetable mixture oils (mean 3.1 %, range 1.1-6.5). CONCLUSIONS: Vegetable oils are the primary home cooking fats in Bogota, Colombia. Higher socio-economic status is associated with usage of sunflower oil. Paradoxically, oblivious to the higher trans content of sunflower oil and the negligible amount of n-3 fatty acids, families commonly reported 'health' as a reason to choose sunflower over other oils. PMID- 18426634 TI - Targets to tackle the obesity epidemic: a review of twelve developed countries. AB - OBJECTIVE: Health targets describe government intentions for improving population health. The present paper determines whether the targets which twelve developed countries have set for obesity match the seriousness of the public health problem. DESIGN: Policy documents on general public health, obesity, nutrition and physical activity were obtained by repeated searches of government websites. Details of all relevant targets on overweight, obesity, nutrition and physical activity were extracted. RESULTS: Only four of the countries studied have set targets for specific reductions in the prevalence of obesity. Two have targets that only mention reducing the prevalence of obesity and two other countries wish to halt the rise in prevalence. Two countries currently have targets which are much less ambitious than those in previous policies. No obesity targets are stated in the policies of four countries. Many of the countries studied have set detailed nutrition targets, but these seldom identify desired changes in dietary behaviour. No country has set targets for a reduction in energy intake. The physical activity targets reflect recommendations from the 1990s. Few targets are set for health knowledge or behavioural intentions which are prerequisites for desired lifestyle changes. CONCLUSIONS: Most of the countries studied have either set no targets or set very modest targets for reducing the prevalence of obesity. Many countries have physical activity targets that are likely to be insufficient to prevent obesity. Governments need to reconsider targets on obesity and to develop shorter-term targets which monitor desired lifestyle changes. PMID- 18426635 TI - The gap between food-based dietary guidelines and usual food consumption in Belgium, 2004. AB - OBJECTIVE: To evaluate the gap between food-based dietary guidelines (FBDG) and the usual food consumption in Belgium. DESIGN AND SETTING: Information on food intake was collected with two non-consecutive 24 h recalls, using the validated software package EPIC-SOFT in combination with a self-administered FFQ. Habitual food intake was estimated by the Nusser method. Physical activity was evaluated according to the International Physical Activity Questionnaire. SUBJECTS: A representative sample of the Belgian population aged 15 years and older was randomly selected from the National Register using a multistage stratified procedure. Dietary information was obtained from 3245 individuals. RESULTS: Food intakes deviated significantly from the recommendations. In particular, fruit (118 g/d) and vegetable (138 g/d) consumption and intake of dairy and Ca-enriched soya products (159 g/d) were inadequate. Consumption of energy-dense, nutrient poor foods (soft drinks, alcohol and snacks) was excessive (481 g/d). There were important age and gender differences. Fruit, vegetable and spreadable fat consumption was lowest, while consumption of dairy, starchy and energy-dense, nutrient-poor foods was highest among the youngest age group. Men consumed more animal and starchy foods than women, who consumed more fruits. There were only slight differences by education level. CONCLUSION: Food intakes differed substantially from the FBDG. Improvement of the Belgian food pattern, in particular among the youngest age group, is necessary for a better prevention of diet-related diseases. In addition, continuous or regular monitoring is crucial to permit trend analyses and to plan effective education or intervention strategies. PMID- 18426636 TI - Setting priorities for zinc-related health research to reduce children's disease burden worldwide: an application of the Child Health and Nutrition Research Initiative's research priority-setting method. AB - OBJECTIVE: To make the best use of limited resources for supporting health related research to reduce child mortality, it is necessary to apply a suitable method to rank competing research options. The Child Health and Nutrition Research Initiative (CHNRI) developed a new methodology for setting health research priorities. To broaden experience with this priority-setting technique, we applied the method to rank possible research priorities concerning the control of Zn deficiency. Although Zn deficiency is not generally recognized as a direct cause of child mortality, recent research indicates that it predisposes children to an increased incidence and severity of several of the major direct causes of morbidity and mortality. DESIGN: Leading experts in the field of Zn research in child health were identified and invited to participate in a technical working group (TWG) to establish research priorities. The individuals were chosen to represent a wide range of expertise in Zn nutrition. The seven TWG members submitted a total of ninety research options, which were then consolidated into a final list of thirty-one research options categorized by the type of resulting intervention. RESULTS: The identified priorities were dominated by research investment options targeting Zn supplementation, and were followed by research on Zn fortification, general aspects of Zn nutrition, dietary modification and other new interventions. CONCLUSIONS: In general, research options that aim to improve the efficiency of an already existing intervention strategy received higher priority scores. Challenges identified during the implementation of the methodology and suggestions to modify the priority-setting procedures are discussed. PMID- 18426637 TI - The role of mentoring in public health nutrition workforce development. Perspectives of advanced-level practitioners. AB - OBJECTIVE: To explore the nature, role and utility of mentoring in the development of competence in advanced-level Australian public health nutritionists. DESIGN: Qualitative study using in-depth interviews. SUBJECTS AND SETTING: Eighteen advanced-level public health nutritionists working in academic and practice settings in Australia. RESULTS: The attributes and career pathways of the subjects were consistent with previous findings. Dissatisfaction with clinical practice was a key reason for choosing a career in public health. Experiential learning, postgraduate education and mentoring from both peers and senior colleagues were the most significant contributors to competency development. The subjects supported mentoring as an important strategy for public health nutrition workforce development and articulated the characteristics and models important for mentoring relationships in public health nutrition. CONCLUSIONS: The present study suggests mentoring was an important part of competency development for advanced-level public health and community nutritionists in Australia. Mentoring programmes based on experiential learning may assist in developing public health nutrition workforce competence. PMID- 18426638 TI - Reduced away-from-home food expenditure and better nutrition knowledge and belief can improve quality of dietary intake among US adults. AB - OBJECTIVE: To test whether reduced away-from-home food expenditure (AFHFE) and better nutrition knowledge and beliefs (NKB) are associated with dietary quality among US adults. DESIGN AND SUBJECTS: The dietary intake data (average of two 24 h recalls) used were collected from US adults (20-65 years) participating in two cross-sectional surveys, the 1994-96 Continuing Survey of Food Intake by Individuals (CSFII; n 7148) and the CSFII/Diet and Health Knowledge Survey (DHKS; n 4252). OUTCOME MEASURES: Dietary quality was assessed using selected nutrients and food groups and the 2005 revised US Department of Agriculture Healthy Eating Index (HEI). EXPOSURE VARIABLES: (i) Absolute AFHFE (weekly, per capita) and proportion of this exposure out of total food expenditure (relative expenditure); (ii) NKB score using a composite of an eleven-item scale elicited among the CSFII/DHKS subgroup. STATISTICAL ANALYSES PERFORMED: We used t tests, chi2 tests, Wilcoxon rank-sum tests and multivariate linear regression models adjusting standard errors for sample design complexity. We utilized a change-in-estimate approach to assess mediation. For effect modification, we tested the significance of interaction terms (NKB x AFHFE). RESULTS: Absolute AFHFE was positively associated with grams of fat (beta = 0.14 (SE 0.06)) and saturated fat (beta = 0.02 (SE 0.01)) and negatively associated with fibre (beta = -0.02 (SE 0.01)) and HEI (beta = -0.08 (SE 0.01)). Relative AFHFE mediated NKB effects on intakes such as fat, saturated fat, cholesterol, Na, and fruits and vegetables (change in estimate >10 %). Among subjects with a poor NKB score, higher AFHFE resulted in lower diet quality, particularly Na and cholesterol intakes. CONCLUSIONS: Higher AFHFE was associated with a lower dietary quality and interacted antagonistically with NKB in some instances, while mediating the relationship between NKB and dietary quality in others. PMID- 18426640 TI - [Multiple myeloma: past, present and future--editorial]. AB - Multiple myeloma is a neoplasm of mature and immature plasma cells, it remains an incurable disease using conventional chemotherapy and increasing aggressive approaches. In recent years, due to the better understanding of myeloma biology, genetics and tumor formation, there are lots of new active drugs or combinational chemotherapy regimens having been developed, such as proteasome inhibitors, immunomodulatory agents etc, they are more effective than conventional chemotherapy. This article summarizes the recent advances with the new options for the treatment of multiple myeloma. PMID- 18426639 TI - Effect of different dosage and administration schedules of folic acid on blood folate levels in a population of Honduran women of reproductive age. AB - BACKGROUND: Observational studies and clinical trials have shown conclusive evidence that periconceptional folic acid supplementation prevents up to 70 % of neural tube defects (NTD). The Honduran government wanted to implement a supplementation programme of folic acid but needed to assess the relative effects of two dosages of folic acid. OBJECTIVE: To determine the effect of two dosages of folic acid on blood folate levels in Honduran female factory workers aged 18 to 49 years. DESIGN: This was a randomized, double-blind control supplementation trial conducted in Choloma, Honduras. A total of 140 eligible women were randomly assigned to two dosage groups and followed up for 12 weeks. One group received a daily dosage of 1 mg folic acid and the other a once weekly dosage of 5 mg. Serum folate and red blood cell folate levels were determined by radioassay at baseline, 6 weeks and 12 weeks. RESULTS: Serum folate levels increased from 6.3 (se 0.2) to 14.9 (se 0.6) ng/ml (P < 0.0001) in women assigned to the 1 mg/d group and from 6.9 (se 0.3) to 10.1 (se 0.4) ng/ml (P < 0.0001) in those assigned to the 5 mg/week group. Red blood cell folate concentrations also increased significantly in both groups, albeit more slowly. Educational level, age and BMI were not associated with the changes in serum and red blood cell folate levels during the supplementation period. However, a differential effect on serum folate levels by dosage group and time was observed. CONCLUSIONS: Although both folate supplementation regimens increased serum and red blood cell folate levels significantly among the women studied, blood folate levels that are considered to be protective of NTD were reached faster with the daily dosage of 1 mg folic acid. PMID- 18426641 TI - [Correlation between DNA methylation of the ABO gene promoter CpG island and leukemia]. AB - Recent studies have found that ABO blood group antigen is also closely related to the onset and development of many diseases. More and more attention is being paid to the decrease of A/B blood group antigen caused by some tumors. This study was purpose to investigate the correlation between DNA methylation of the ABO gene promoter CpG island and leukemia. The relative contents of ABH antigen on the surface of RBC from kinds of blood disease patients and healthy individuals were detected by using flow cytometry and confocal laser scanning microscopy. The DNA sequences and CpG methylation of ABO gene promoter in patients with hematopathy and healthy individuals, as well as the -102 site methylation of ABO gene promoter in patients with hematopathy and healthy individuals were detected by PCR and MSP-PCR respectively. The results showed that RBC from leukemia patients displayed different degree of A/B antigen decrease. The sequences of ABO gene promotor of patients with hematopathy were not different from healthy individuals indicating high conservation of promoter sequences. Comparison of sequences between patients with hematopathy and healthy individual indicated that CpG islands on ABO gene promoter either from blood disease patients or from healthy individual had no methylated site in AA patients, but C residues at position 102, -101, -100, -99 and -97 on the promoter of ABO gene in AML, CML, ALL and some MDS patients were methylated. It is concluded that methylation of CpG islands in promoter of ABO gene may result in AB antigen decrease in patients with leukemia. The methylation sites -102, -101, -100, -99 and -97 may be specific for leukemia. The methylation of site -102 can be used as a molecular marker in differential diagnosis for leukemias. PMID- 18426642 TI - [CD147 increases invasiveness of U937 cells through regulation of matrix metalloproteinase activity]. AB - This study was purpose to investigate the effects of CD147 on the invasiveness of leukemia cells U937. The experiments were divided into 4 groups: control group, LPS group, CD147mAb group and LPS+CD147 mAb group. Cells were treated by lipopolysaccharide (LPS) or anti-CD147 monoclonal antibody, and the expression of CD147 and MMP-2, -9, the invasive potential of the cells in vitro and ex vivo, as well as the invasion of the implanted tumors in SCID mice were analysed by RT PCR, FCM, gel zymography and invasion test in vitro respectively. The results showed that the expression of CD147 was elevated by the induction of LPS, and the enhanced expression of CD147 on U937 cells increased the production and secretion of MMP-2 and MMP-9 as measured by reverse transcription-PCR and gel zymography. An increased number of LPS-induced cells invading through a reconstituted basement membrane were observed by invasion assays. These responses were down regulated after blocking CD147 with anti-CD147 antibody. At 30 days after intravenous injection of LPS pretreated U937 cells to SCID mice human U937 cells were found in the bone marrow and lung of the mice, indicating the invasion of the tumor cells. And overexpressions of CD147, MMP-2 and MMP-9 were found in the lung tissue of the mice injected with LPS-treated but not anti-CD147 antibody treated tumor cells. It is concluded that overexpression of CD147 on U937 cells may increase the secretion and activation of MMP-2 and MMP-9 and thus promote the invasiveness of the tumor cells. PMID- 18426643 TI - [Effect of down-regulating mll-af9 gene expression on proliferation of acute monocytic leukemia cell line THP-1]. AB - This study was aimed to investigate the effect of small interfering RNA (siRNA) on the expression of mll-af9 oncogene and the proliferation of human acute monocytic leukemia cell line THP-1. One group of siRNA was designed targeting mll af9 mRNA and finally obtained by chemosynthesis. Then the obtained siRNA was transfected into cultured human acute monocytic leukemia cell line THP-1 by lipofectamine. Flow cytometry was used to detect siRNA transfection efficiency. The level of mll-af9 mRNA expression was analyzed by reverse transcription polymerase chain reaction (RT-PCR). The cell proliferation rate was assayed by MTT. The change of cell cycles and apoptosis rate was detected by flow cytometry. The results showed that the siRNA transfection efficiency was 69.1%+/-1.8%. The level of mll-af9 mRNA expression was significantly inhibited in siRNA-transfected cells as compared with the controls. mll-af9-targeted siRNA inhibited the proliferation of THP-1 cells and induced cell apoptosis effectively after transfection. The percentage of G0/G1 phase cells significantly increased in siRNA-transfected cells in comparion with the control cells, but the percentage of S phase cells significantly decreased. It is concluded that the mll-af9 targeted siRNA can effectively inhibit the proliferation of human acute monocytic leukemia cell line THP-1. PMID- 18426644 TI - [Construction of kir3dl1 promoter expression vector and its activity in K562 cell line]. AB - To analyze the function of kir3dl1 core promoter and study the possible regulation mechanism of kir3dl1 gene expression, a kir3dl1 promoter-luciferase reporter vector was constructed and the promoter activity was evaluated in the K562 cell line. A core promoter fragment of the kir3dl1 5'-untranslated region was amplified by PCR. PCR products were cloned into BglII/NcoI-digested pGL3 basic reporter vector; the polycationic compound SuperFect-reporter vector complexes were transferred into K562 cells. The Dual-Luciferase Reporter Assay System was used to quantitate the reporter vector luciferase activity. MTT method was used to measure the influence of SuperFect-DNA complexes on the survival rate of K562 cells. The results indicated that a 254-bp promoter fragment of kir3dl1 gene was successfully constructed and cloned into the pGL3-basic reporter vector, which was authenticated by BglII/NcoI digestion and DNA sequencing. The luciferase activity of the minimal promoter construct was significantly higher than that of the pGL3-Basic promoter in K562 cells. Transiently transfected cells presented continuously optimal luciferase activity and relative luciferase activity up to 3 days. The cell activity was between 76% and 92%. It is concluded that a kir3dl1 promoter-luciferase reporter vector is successfully constructed, the transfection system used in this study can effectively transfer gene in K562 cells. The kir3dl1 core promoter possesses higher activity in K562 cells, and can promote significantly expression of luciferase reporter gene in K562 cells. PMID- 18426645 TI - [Significance of DNA methylation status of runx3 gene promoter region in acute leukemia]. AB - This study was aimed to investigate the effects of methylation of runx3 gene promoter on pathogenesis of acute leukemia (AL) and its clinical significance. The methylation of runx3 gene promoter in cells of bone marrow or peripheral blood from 40 cases of AL and 10 healthy persons as well as in CHRF, U937 and K562 cell lines were assuaged by methylation specific polymerase chain reaction (MS-PCR), the expression of runx3 gene were detected with reverse transcription polymerase chain reaction (RT-PCR). The results indicated that no methylation was detected in all of cell lines and healthy persons while expression of runx3 gene could be deteted, methylation of runx3 gene promoter was found in 35% (14/40) AL patients and its percentage was significant higher than that healthy persons (0%), the difference in methylation for runx3 between two kinds of samples was statistically significant (p<0.05), while methylation rate in AML was 30.43% (7/23), ALL was 41.18% (7/17), there was no significant difference between them (p>0.25). All of methylated samples had no expression of runx3 gene. Patients without methylation of runx3 gene had a lower percentage of blasts in bone marrow and a higher complete remission rate of first chemotherapy than those with methylation of runx3 gene. It is concluded that methylation of runx3 gene promoter probably plays a role in the pathogenesis of AL and may have clinical significance in predicting prognosis of AL. PMID- 18426646 TI - [Expression characteristics of heat shock protein 70 and pim-1 gene in bone marrow mononuclear cells from leukemia patients and their clinical significance]. AB - This study was aimed to investigate the expression characteristics of HSP70 protein/mRNA, pim-1 mRNA in bone marrow mononuclear cells from leukemia patients, and to clarify whether these changes are related to leukemia type, tumor burden of leukemia, therapeutic reaction and prognosis. HSP70 mRNA and pim-1 mRNA in BMMNCs were detected with semi-quantitative RT-PCR in 40 leukemia patients and 10 controls. HSP70 protein in BMMNCs was assayed with Western blot in 34 leukemia patients and 10 controls. Relation of HSP70 and pim-1 expression with leukemia classification, the degree of tumor burden and therapeutic reaction were analyzed. The results showed that the BMMNCs from both leukemia patients and controls expressed HSP70 protein/mRNA. The mean ODR value of HSP70 mRNA in BMMNCs from leukemia patients was significantly higher than that of the controls; the mean ODR value of HSP70 protein/mRNA in acute myeloid leukemia and chronic myeloid leukemia patients both were significantly higher than that of acute lymphocytic leukemia patients; the mean ODR value of HSP70 protein/mRNA in acute leukemia patients with high-degree tumor burden was higher than that of the patients with low-degree tumor burden; the mean ODR value of HSP70 protein/mRNA in the patients after chemotherapy was significantly higher than that of the patients before chemotherapy; the BMMNCs from both leukemia patients and controls expressed pim-1 mRNA. The mean ODR value of pim-1 mRNA in BMMNCs from leukemia patients was significantly higher than that of the controls; the mean ODR value of pim-1 mRNA in BMMNCs for Acute lymphocytic leukemia patients was significantly higher than that of the patients suffered from acute myeloid leukemia and chronic granulocytic leukemia; there was a positive relationship between pim-1 mRNA and HSP70 mRNA expressions in leukemia patients (p < 0.05). It is concluded that there are high expressions of HSP70 and pim-1 in leukemia and their positive correlation is shown. The over-expressions of HSP70 and pim-1 protein/mRNA are related to tumor burden in leukemia patients. PMID- 18426647 TI - [Effect of CTL on K562 cell induced by exosomes and in combination with CPG OND]. AB - To investigate the specific anti-leukemia effect of cytotoxic T lymphocytes (CTLs) induced by dendritic cells (DCs) activated by exosomes alone or in combination with CpG ODN in vitro and the feasibility of exosomes as remedial vaccine, the DCs induced from normal volunteer PBMNCs were divided into 7 groups. Three groups of them were added with the exosomes: Kexo (exosomes derived from K562 cells) or DCexo (exosomes derived from DCs induced from K562 cells) or FTexo (exosomes derived from DCs induced from K562 cells and pulsed by freeze-thawing antigen of K562 cells) as experimental groups (Kexo, DCexo and FTexo). The other three groups were added with CPG ODN while added the exosomes (Kexo, DCexo and FTexo), and were used as experimental groups also (Kexo+CpG, DCexo+CpG and FTexo+CpG). The seventh group DCs was added with nothing as blank control. These DCs above mentioned were cultured continuously for 72 hours. The T lymphocytes were co-cultured with DCs for another 72 hours to generate CTL. Then, the killing effects of them on K562 cells were determined by MTT assay. The results showed that all experimental groups pulsed by exosomes displayed stronger killing effect, compared with control group (p<0.05). DCexo and FTexo displayed stronger killing effect too, compared with Kexo (p<0.05). CPG ODN as an adjuvant could enhance the killing effect (p<0.05). It is concluded that the special killing effect on K562 cells can be induced by exosomes, CPG ODN as an adjuvant can enhance the killing effect. Exosome is hopeful as a remedial vaccine to be used for the leukemia therapy. PMID- 18426648 TI - [Synergistic antileukemic effect of phytoestrogens and chemotherapeutic drugs on leukemic cell lines in vitro]. AB - Natural phytoestrogens such as the isoflavones genistein and daidzein, and the flavones quercetin exhibit anti-cancer properties. This study was purpose to investigate the anti-proliferative effect of phytoestrogens on acute myeloid leukemia (AML) and chronic myeloid leukemia (CML) cells, and their synergistic antileukemic effect in combination with chemotherapeutic drugs. Optimal dosage of genistein, quercetin and in combination with chemicals for leukemia cells were determined by experiments. Cell viability, apoptosis induction and cell cycle arrest were detected by trypan blue staining, MTT assay, optical microscopy, flow cytometry (FCM). The schedule treatment of combination of genistein and chemicals was determined. The results showed that genistein exhibited a dose- and time dependent inhibitory effect on cell proliferation in NB4 and HL-60 cells, induced apoptosis and cell cycle arrest in G2/M phase. Quercetin had evident inhibitory effect on the proliferation of K562 and K562/A cells. The combination of genistein and chemicals exerted a synergistic effect on cell growth inhibition. In conclusion, this study demonstrated the synergistic antileukemic effect of genistein with chemotherapeutic drugs on leukemic cells. This combination appears to be a new idea for the clinical novel treatment of leukemia. PMID- 18426649 TI - [Expression spectra of apoptosis-related gene pnas-2]. AB - To explore the expression spectra of apoptosis-related gene pnas-2 in normal tissues and acute leukemia (AL) patient tissues, the expressions of pnas-2 gene in tissues including heart, brain, placenta, lung, liver, skeletal muscle, kidney, pancreas, spleen, lymph node, thymus, leukocyte, bone marrow and fetal liver were detected by Northern blot. The expressions of pnas-2 in samples including 44 de novo, 9 non-CR, 27 CR and 12 relapsed AL patients were measured by real-time RT-PCR and Northern blot, and the expression levels of pnas-2 in normal and tumor tissues from 31 patients with malignancies were also detected. The results showed that pnas-2 was not expressed in the most tissues except in placenta. The results of real-time PCR indicated that pnas-2 expressions in samples of de novo, non-CR and relapsed patients ware significantly higher than that in CR, tumor tissues and normal tissues. In serial monitoring of 7 AL patients, the expression level of pnas-2 was high at first visit examination, but remarkably decreased after remission, and the pnas-2 expression level increased again when relapsed. It is concluded that the pnas-2 is specifically up-regulated in acute leukemia patients, which might be an oncogene and participate in leukemogenesis. PMID- 18426650 TI - [Relation of apoptosis of K562 cells induced by naringenin in vitro to enzyme activity changes of caspase-3 and caspase-8 and expression of FAS/FASL proteins]. AB - The objective of this study was to investigate the apoptosis-inducing effect and underlying mechanism of naringenin (NGEN) on K562 cells in vitro. The inhibition of NGEN on K562 cells was evaluated by means of MTT assay so as to observe the cytotoxicity of NGEN; The morphological changes of the cells treated by NGEN were observed by transmission electron microscope; cell apoptosis rate influenced by NGEN was assessed by flow cytometry; the enzyme activity changes of caspase-3 and caspase-8 in the process of NGEN-induced K562 apoptosis were detected by Caspase Colorimetric Assay Kit; immunohistochemistry technique was used to detect FAS, FASL protein expression in K562 cells. The results showed that the growth of cells was inhibited by NGEN in dose-and time-dependent manners (p<0.05); NGEN induced K562 cells apoptosis and sub-G1 peak was observed; some typically early and final phase changes of cell apoptosis were revealed under transmission electron microscope; the enzyme activity of caspase-3 and caspase-8 and the expression of FAS remarkably increased, meanwhile the expression of FASL was down regulated (p<0.05). It is concluded that NGEN exerts anti-cancer effect by inducing K562 cell apoptosis, and the regulation of the expression of FAS and FASL. The caspase-3 and caspase-8 co-pathway brings about one of the mechanisms. PMID- 18426651 TI - [Apoptosis of HL-60 cells induced by aescinate]. AB - The aim of this study was to investigate the effects of aescinate on inhibition and apoptosis of HL-60 cell line from promyelocytic leukemia. HL-60 cells at logarithm phase were treated with aescinate. Cell survival rate and cell morphology were observed, and the cell apoptosis was analyzed by Annexin V/PI FITC double labeling and DNA electrophoresis. The results showed that HL-60 cells could be inhibited in the presence of 15-120 mg/L of aescinate for 48 hours, survival rates were (92.2+/-0.69)%-(8.2+/-0.96)%, which were significantly lower than that of non-aescinate control (99.4+/-0.31)% (all p<0.01). The apoptosis of cells could be induced by aescinate treatment at dosage of 15-60 mg/L for 24 hours, the Annexin V positive cells accounted for (12.7+/-0.58)%-(65.4+/-1.30)% which were significantly higher than that of non-aescinate control (0.57+/-0.03)% (all p<0.01). The typical DNA ladder of HL-60 cells treated with aescinate was shown on the DNA electrophoresis pattern. It is concluded that aescinate can specifically induce apoptosis of leukemic HL-60 cells, which provides an experimental evidence for treatment of leukemia with aescinate as a supplementary agent to chemotherapy. PMID- 18426652 TI - [Expression of human telomerase reverse transcriptase and survivin gene in patients with myelodysplastic syndrome]. AB - This study was aimed to investigate the expressions of human telomerase reverse transcriptase (hTERT) and survivin gene in patients with myelodysplastic syndrome (MDS), and to explore their relationship. The expression of hTERT mRNA in bone marrow mononuclear cells (BMMNCs) of 56 patients with MDS and 27 patients with iron deficiency anemia were detected by RT-PCR, the expressions of survivin gene in BMMNCs of 55 patients with MDS and 12 patients with iron deficiency anemia were detected by real-time RT-PCR. The results showed that the expression of hTERT significantly elevated in RA and RAEB patients, as compared with controls (p<0.005). With the disease alleviated, the expression of hTERT decreased and had no significant difference from the controls (p>0.25). There was no significant difference in expression of hTERT between low+int-1 risk group and int-2+high risk group by IPSS (p>0.50). The expression of survivin gene significantly increased in RA and RAEB patients, as compared with controls (p<0.02, p<0.05). The expression of survivin gene in low+int-1 risk group by IPSS was significantly higher than that in the controls (p<0.02), and there was no significant difference in expression of survivin gene between int-2+high-risk group patients and the controls (p>0.10). It is concluded that the expressions of hTERT and survivin may play a critical role in escaping malignant clone of MDS from apoptosis and acquiring the ability to divide unlimitedly. PMID- 18426653 TI - [Immuno-suppressive effects on T cells mediated by mesenchymal stem cells from patients with myelodysplastic syndrome]. AB - The study was aimed to compare the effects of T-cell suppression mediated by mesenchymal stem cells (MSC) from normal individuals and myelodysplastic syndromes (MDS) patients. MSC were cultured from the bone marrow of 12 healthy volunteers and 12 MDS patients, the morphology, surface markers and expression of several cytokines of MSC from normal individuals and MDS patients were compared, and the effects of T-cell suppression were tested in the following assays: phytohemaglutinin (PHA)-primed cultures, mixed lymphocyte reaction (MLR), cell cycle of T-cell after PHA-primed cultures and apoptosis of T-cell as well. The results showed that the MSC from normal individuals and MDS patients were similar in morphology, proliferation and surface markers. The suppressions of T-cell proliferation induced by PHA and alloantigens mediated by MDS-MSC were significantly lower than that of normal MSC. More T-cells were arrested in G0/G1 phase by normal MSC, while the effects were deficient by MDS-MSC. The suppression of T-cell activation mediated by MDS-MSC was also lower than that of normal MSC, but suppression effect on T-cell apoptosis increased. The cytokines TGF-beta1, 3, FasL expressed by MDS-MSC were reduced as compared with normal MSC, but TGF-beta2 expression increased in MDS-MSC. It is concluded that although the morphology, proliferation and cell surface markers of MDS-MSC are normal, the T-cell suppression mediated by MDS-MSC is deficient as compared with normal controls. Whether these abnormalities are relevant to the pathogenesis of aplastic anemia remains to be determined. PMID- 18426654 TI - [Scoring analysis on prognosis of 63 patients with melodysplastic syndrome]. AB - The purpose of this study was to investigate the risk factors in prognosis of patients with myelodysplastic syndrome (MDS) to emphatically study the clinical significance of different scoring systems such as Bournemouth-, Spanish-, Dusseldorf-, Lille, and the International prognostic scoring systems (IPSS), French-American-British (FAB) and World Health Organization (WHO) classifications as well as abnormal clone of chromosome, bone marrow and hematological indicators in evaluating prognosis of MDS patients, and to identify the independent factor related to prognosis. All clinical data of 69 consecutive patients diagnosed as primary MDS over a period of 5 years were collected and stored by Epi Data 3.0, 63 patients out of which were followed up. SPSS 13.0 software was applied to deal with all data. The statistical methods included life table, Kaplan-Meier, Log rank test and Cox regression. The results indicated that the median age of 63 patients was 38 years. 26 out of 63 patients had karyotype aberration (41.27%). Median OS was 30.63 months, and 33 patients (52.38%) died. All five prognostic scoring systems could successfully discriminate risk groups as regards overall survival. IPSS, Lille and Spanish prognostic scoring system were more effective (p<0.0001). Multivariate Cox regression analysis indicated that IPSS (p<0.0001) and Lille chromosome classifications (p<0.0001) were most important factors for OS followed by bone marrow blasts (p=0.00062), Spanish prognostic scoring system (p=0.00064) and Lille prognostic system (p=0.008). The WHO classifications also successfully discriminated between risk groups (p<0.0001). The new WHO subgroups [refractory cytopenia with multilineage dysplasia (RCMD), with or without ringed sideroblasts] showed a significantly different prognosis (p=0.003) for OS, in comparison to the subgroups having erythroid dysplasia only (RA/RARS) and 5q syndrome. All patients were reclassified to FAB classification, and the low risk group (RA/RAS) and high risk group (RAEB) also had significant difference (p=0.00012) as regards OS. It is concluded that the major independent prognostic variables for OS are percentage of bone marrow blasts and karyotype aberration. The use of WHO classification have more significance for improving predictive value than that of the FAB classification, and the IPSS can be used for clinical decision-making in patients with cytogenetic results. In the hospitals which cannot carry out the cytogenetic examination, Spanish prognostic system can be applied to the patients without cytogenetic results. PMID- 18426655 TI - [Effect of VEGF antisense RNA on inducing apoptosis of myeloma cells and inhibition of angiogenesis in endothelial cells in vitro]. AB - This study was aimed to investigate the effect of VEGF antisense RNA on proliferation and apoptosis in myeloma cell line U266 as well as on angiogenesis in endothelial cell ECV304, and to explore the feasibility of gene therapy for multiple myeloma using VEGF antisense RNA. The VEGF121 cDNA was inserted into multiple clone site of eukaryotic expression vector pIRES2-EGFP to construct the recombinant plasmid AS-VEGF. Restriction endonuclease analysis and DNA sequencing were used to confirm the reverse orientation of the VEGF cDNA. The recombinant plasmid was transfected into human myeloma cell line U266 and the positive clone was screened by G418. The VEGF mRNA and protein expressions of the positive clone were detected by RT-PCR and Western blot respectively. The viability and apoptosis of U266 cells were observed by MTT assay, flow cytometry. Angiogenesis was tested by network formation of endothelial cells on matrigel. The results indicated that the recombinant plasmid AS-VEGF expressing VEGF antisense RNA were constructed successfully. VEGF expression in U266 cells was blocked partially by VEGF antisense RNA. Expression of VEGF mRNA and protein decreased more significantly in U266 cells transfected by AS-VEGF than that in control group. Then increasing of apoptosis and inhibition of proliferation in U266 cells transfected by AS-VEGF were observed. Vasoformation on matrigel in the supernatants of U266 culture group transfected by AS-VEGF decreased more significantly than that in control group. It is concluded that VEGF antisense RNA can inhibit the expression of VEGF gene in U266 cells, thereby inhibits the proliferation of U266 cells; increases the apoptosis of U266 cells; and inhibits angiogenesis in vitro. PMID- 18426656 TI - [Synergetic effect of B7-1 and CD40L in the immunotherapy for lymphoma]. AB - The aim of study was to investigate the synergetic effect of B7-1 and CD40L co stimulating pathway in the immunotherapy for lymphoma and to explore the effective manner of tumor vaccine for treating lymphoma. The lymphoma cell line A20 cells were inoculated into BALB/c mice as to establish A20-bearing mice model, the B7-1 and CD40L expression vector were alone or in combination directly injected into lymphoma of mice model, the PBS, vector pcDM8 and pcDNA3.1 were selected as controls so as to observe tumor growth. The pathological section and HE staining of tumor tissue were performed to observe the histological characteristics and the cell infiltration of lymphoma, the CCK-8 detection kit was used to analysis the splenic CTL cytotoxicity. The results showed that the intratumor injection of B7-1 and CD40L resulted in reduction of tumor size. Morphological observation of tumor revealed inflammatory cell infiltration in the tumors, massive necrosis and localization of tumor. CCK-8 kit detection indicated significant enhancement of splenic CTL cytotoxicity, the effect of B7-1 combined with CD40L was stronger than that of B7-1 or CD40L alone. It is concluded that B7 1 and CD40L show immunotherapeutic effect on lymphoma, and the effect becomes stronger when they are combined in treating lymphoma. Meanwhile, the intratumor injection may be considered as a safe and effective way for tumor vaccine. PMID- 18426657 TI - [Significance of id4 gene promoter methylation detection in lymphoma]. AB - The aim of this study was to evaluate the significance of id4 gene promotor methylation detection in NHL patients. MS-PCR method was used to detect the status of id4 gene methylation in health donors and newly diagnosed NHL patients. The results indicated that the id4 gene was unmethylated in bone marrow samples from health donors. Among 18 newly diagnosed NHL patients, including one NHL patient with bone marrow cells involved, 4 patients were found in id4 gene methylation by MS-PCR. The 14 patients with id4 gene unmethylation were in their stable status and no bone marrow involvement were found by bone marrow biopsy during the 8-month follow-up. During the follow-up, the patient with both bone marrow involvement and id4 gene methylation turned to leukemia, in 2 out of the 3 patients with id4 gene methylation but without bone marrow involvement at diagnosis, the bone marrow involvement was found at last. It is concluded that the id4 gene methylation may be an indicator for MRD in NHL patients without bone marrow involvement. PMID- 18426658 TI - [Single nucleotide polymorphism of interferon-gamma gene +874 T/A in patients with aplastic anemia]. AB - The aim of this study was to investigate whether the single nucleotide polymorphism (SNP) of interferon-gamma (IFN-gamma) +874 T/A correlates with aplastic anemia. Amplification refractory mutation system-polymerase chain reaction was used to amplify the polymorphism gene segment of IFN-gamma +874 A/T from 54 aplastic anemia patients and 51 healthy adults. The results showed that the frequency of IFN-gamma +874 TT genotype and T allele was significantly higher in patients with aplastic anemia than that in the healthy adults (42.6% vs 17.6%, chi2=13.780, p=0.01; 65.7% vs 39.2%; chi2=14.811, p<0.001). In conclusion, IFN gamma +874 A/T gene polymorphism is correlated with the susceptibility of aplastic anemia, but not significantly correlated with the the severity of aplastic anemia. PMID- 18426659 TI - [Expression of human ermap gene in umbilical cord blood mononuclear cells during differentiation and development towards erythroid lineage]. AB - The aim of study was to explore the potential of human erythroid membrane associated protein (ERMAP) gene in erythroid cell differentiation and development, mononuclear cells (MNCs) were isolated from umbilical cord blood and induced to erythroid cell differentiation by SCF, IL-3 and EPO. The cell morphology was observed by using optical microscopy, the positive rate of cells was counted by biphenylamine staining and the ratios of CD36+/CD235a-, CD36+/CD235a+, CD36-/CD235a+ cells were detected by flow cytometry, the change of human ermap gene expression level was analyzed by using fluorescent quantitative PCR (FQ-PCR). The results showed that the ermap gene expression level increased while MNCs were induced to erythroid lineage after treatment with SCF, IL-3 and EPO. It is concluded that the human ermap gene plays an important role in differentiation and development of erythroid cells. PMID- 18426660 TI - [Comparison between CD271 and CD133 used for immunomagnetic positive sorting enriching in mesenchymal stem cells from bone marrow]. AB - This study was aimed to find a better method to isolate and enrich mesenchymal stem cells (MSCs)from bone marrow between CD271 (low affinity nerve growth factor receptor, LNGFR) and CD133 used for immunomagnetic positive selections through comparison of characteristics of MSCs isolated by these two agents. CD271+ and CD133+ cells were isolated from bone marrow and their colony forming unit fibroblast (CFU-F) efficiency and proliferative capacity were assessed. Cell surface phenotype, adipogenic and osteogenic inductions were also assayed on the cells (after passage 3) isolated by both methods. The results showed that the purities of immunomagnetically selected CD271+ and CD133+ cells were (89.50+/ 0.98)% and (88.03+/-3.06)% respectively. The CFU-F median frequency of CD271+ cells was 3 times as high as that of CD133+ cells, no CFU-F was observed in CD271 cells, while a few CFU-F was found in the CD133- cells. Phenotype of cells (after passage 3) isolated by the two methods was same, that is CD34-, CD14-, CD45-, CD90+, CD29+, CD44+, CD105+, CD73+. CD271+ cells possessed faster proliferation and stronger osteogenic and adipogenic differentiation potential than that of CD133+ cells. It is concluded that as compared with CD133 positive selection, CD271 positive selection is a better method for isolating and enriching mesenchymal stem cells from bone marrow. PMID- 18426661 TI - [Biological characteristics of human fetal osteoblastic 1.19 cell line]. AB - This study was aimed to investigate the biological characteristics of osteoblasts and their hematopoietic supportive function by using human fetal osteoblastic cell line 1.19 (hFOBs) as a model. The pluripotency markers (Oct-4, Rex-1, hTERT) of hFOBs were analyzed by RT-PCR, the multilineage differentiation experiments were conducted in vitro. Flow cytometry (FCM) was used to identify the surface markers of hFOBs, and RT-PCR was used to analyze their hematopoietic cytokine expression in comparison with bone marrow mesenchymal stem cell (BM-MSC). The results showed that hFOBs expressed several ESC pluripotency markers including Oct-4 and Rex-1, except hTERT. Moreover, hFOBs could also undergo multilineage differentiation into the mesodermal lineages of adipocytic cell types in addition to its predetermined pathway, the mature osteoblast. Both hFOBs and BM-MSC expressed CD44, CD73 (SH3), CD105 (SH2) and CD90 (Thy1), and lack expression of CD34, CD45, or HLA-DR surface molecules. In addition, both hFOBs and BM-MSC expressed SCF, IL-6, and SDF-1alpha mRNA, but only hFOBs could express GM-CSF and G-CSF. It is concluded that human fetal osteoblastic cell line 1.19 may provide a good model to study the osteoblastic regulation role in hematopoiesis in vitro. PMID- 18426662 TI - [Effects of recombinant human interleukin 11 and granulocyte colony stimulating factor in mobilization for autologous peripheral blood stem cell transplantation]. AB - This study was aimed to evaluate the effects of recombinant human interleukin 11 (rhIL-11) and recombinant human granulocyte colony stimulating factor (rhG-CSF) in mobilization for autologous peripheral blood stem cell transplantation (APBSCT). 16 patients with non-Hodgkin's lymphoma or acute myeloblastic leukemia were given myelosuppressive chemotherapy, then were mobilized by using rhG-CSF 5 microg/(kg.d) for median 5.5 days and rhIL-11 50 microg/(kg.d) for median 4 days (experimental group) or rhG-CSF 5 microg/(kg.d) alone for median 5.5 days (control group). After mobilizing, the peripheral blood leucocyte and platelet counts, total mononuclear cells, CD34+ cells and CFU-GM counts in PBSC collection, and amount of apheresed platelet transfusion were assayed. The results showed that the peripheral blood leucocyte and platelet counts, total mononuclear cell, CD34+ cell and CFU-GM counts in PBSC collection were no significant difference between two groups (p>0.05). After APBSCT, the median time for neutrophil count>or=0.5x10(9)/L and the median time for platelet count>or=20x10(9)/L were 10.5 and 11.5 days in experimental group, while were 13 and 13 days in control group, respectively. The median amount of apheresed platelet transfusion was 3.5 unit in experimental group and 5 unit in control group. Data were significantly different between two groups (p<0.05). The adverse reactions of mobilization were mild fever, fatigue, symptoms like as common cold, poor appetite, dizziness, muscular soreness in experimental group, but only mild fever in control. These symptoms were well tolerated and overcome with drug withdrawal. It is concluded that the regimen of rhIL-11 in combination with rhG CSF after myelosuppressive chemotherapy to mobilize PBSC is efficient and safe with rapid hematologic reconstitution and less platelet transfusions after APBSCT were used. PMID- 18426663 TI - [Effects of IL-21 alone or in combination with IL-15/IL-2 on proliferation and anti-tumor activity of G-CSF-mobilized peripheral blood mononuclear cells in vitro]. AB - This study was purposed to investigate the proliferation and antitumor activity of rhG-CSF-mobilized peripheral blood mononuclear cells (G-PBMNCs) activated by interleukin 21 (IL-21) alone or in combination with interleukin 15 (IL 15)/interleukin 2 (IL-2) and to evaluate the feasibility and value of tumor immunotherapy with cytokine combinations. G-PBMNCs were activated by IL-21 alone or in combination with IL-15/IL-2 in vitro, and the proliferation of the activated G-PBMNCs was analyzed by CCK-8 assay. The cytotoxicity of the activated G-PBMNC to the K562 cells was studied by the test principle which is based on target cell labeling with 5-(6)-carboxy-fluorescein succinimidyl ester (CFSE) and subsequent DNA-labeling with propidium iodide (PI) for identification of target cells with compromised cell membranes. The phenotypes of the activated G-PBMNCs were assayed by flow cytometry. The results showed that the cytotoxicity of IL-21 group had no difference from which of IL-2 group. When G-PBMNCs were exposed to the combinations of IL21+IL15/IL21+IL15+IL2, the cytotoxicity was significantly enhanced at E:T ratio of 25:1, as compared with combination of IL21+IL2 (p<0.05). The cytotoxicity of the cytokines combinations was significantly higher than that in cytokine used alone at E:T ratio of 50:1 (p<0.05). The cryopreservative and resuscitative G-PBMNCs showed the same result with the fresh G-PBMNCs in cytotoxicity test. The proportions of CD3+ and CD8+ T cells were increased when G PBMNCs were incubated with cytokines for 72 hours. CD4, CD3-56+ and CD3+56+ counts were significantly elevated when G-PBMNCs were exposed to IL21 + IL15 (p<0.05). It is concluded that IL-21 alone enhance the antitumor activity of G PBMNCs, which further strengthens when IL-21 combinated with IL-15. PMID- 18426664 TI - [Hematopoietic reconstitution by co-transplantation of human BM-MSCs and UCB CD34+ cells at various times in NOD/SCID mice]. AB - To investigate the effect of co-transplantation of bone marrow derived MSCs and UCB CD34+ cells at different time points on hematopoietic reconstitution, all NOD/SCID mice were sublethally exposed to irradiation of 60Co gamma ray and transplanted with UCB CD34+ with or without MSCs (3 mice per group). Animals were divided into HSC group and MSC+HSC group (M+H group). In HSC group, 1x10(6) UCB CD34+ cells for each mouse were infused within 4-6 hours after irradiation; the M+H group again was divided into 3 subgroups according to infusion sequence of MSCs and HSCs. (A) M+H simultaneously infused group: MSCs and UCB CD34+ cells were infused simultaneously; (B) M+48H group: MSCs were infused within 4-6 hours after irradiation, while UCB CD34+ cells were infused at 48 hours after irradiation; (C) H+48M group: UCB CD34+ cells were infused within 4-6 hours after irradiation, while MSCs were infused at 48 hours after irradiation. In 3 subgroups infused amounts of MSCs and UCB CD34+ cells all were 1x10(6) cells. From the 3rd day after transplantation, 20 microl peripheral blood was collected from the retro-orbital plexus of mice every week until 42th day after transplantation. 42 days after transplantation, mice were sacrificed, and the percentages of human CD45, CD34, CD19 and CD11b in bone marrow, peripheral blood and spleen were detected by FACS. The results showed that (1) Co-transplantation of MSCs and UCB CD34+ cells simultaneously (M+H group) can mitigate the decrease of WBC and platelet levels (p<0.01) in peripheral blood, and accelated the hematopoietic recovery. While co-transplanting MSCs and UCB CD34+ cells at different time points (M+48H or H+48M), the similar effect was not observed (p>0.05). As far as platelets was concerned, the recovery of platelets in M+48H group was lagged behind that in M+H group (p<0.01). (2) Co-transplantation of MSCs at different time points enhanced the engraftment of hematopoietic cells (p<0.05 or p<0.01), compared with transplantation of CD34+ cells alone. The effect of engraftment enhancement was not lineage restriction (p>0.05). It is concluded that the ideal transplantation effect is achieved when MSCs and UCB CD34+ cells were co-transplanted at the same time, these study results provide experimental basis for clinical application of MSCs. PMID- 18426665 TI - [Change of IL-4 and IL-6 levels in blood plasma of patients transplanted with allogeneic hematopoietic stem cells determined by protein chip and its relationship with aGVHD]. AB - This study was aimed to explore the relationship between changes of IL-4, IL-6 levels in blood plasma of patients receiving allo-HSCT and aGVHD through dynamical detection. The IL-4 and IL-6 levels in peripheral blood were detected by protein chip and were observed for 10 weeks. The results indicated that the IL 4 level increased rapidly in 1 week after transplantation in aGVHD group and was higher than that of non aGVHD group (p<0.05), while at 7th week after transplantation IL-4 level increased rapidly in non-GVHD group and was higher than that of aGVHD group (p<0.01). There was a significant difference of IL-6 level between these two groups before transplantation. IL-6 level reached peak at 1st week after transplantation in aGVHD group and was higher than that of non aGVHD group (p<0.01), while IL-6 level was significantly higher in non-aGVHD group than that of aGVHD group at 4th week after transplantation (p<0.01) and at 5th week after transplantation (p<0.05). It is concluded that the levels of IL-4 and IL-6 had been increased rapidly after hematopoietic stem cell transplantation, indicating that aGVHD will occur. The high level of IL-6 before transplantation may be the risk factor of aGVHD occurrence after transplantation. aGVHD may occur later if the blood plasma IL-4 level rises in patients without aGVHD. Therefore, dynamically monitoring IL-4 and IL-6 levels contributes to predict the occurrence of aGVHD. PMID- 18426666 TI - [Pure red cell aplasia following ABO-incompatible allogeneic hematopoietic stem cell transplantation]. AB - The objectives of study was to investigate the clinical characteristics and risk factors of pure red cell aplasia (PRCA) following ABO-incompatible allogeneic hematopoietic stem cell transplantation (allo-HSCT). The clinical data of 72 patients receiving ABO-incompatible allo-HSCT were collected and retrospectively studied. The clinical parameters including sex, age, granulocyte engraftment time and blood transfusion were analyzed for the exploration of risk factors resulting in development of PRCA. The results indicated that 4 out of 72 patients receiving ABO-incompatible allogeneic hematopoietic stem cell transplantation developed PRCA, 3 cases out of these patients were ABO-major incompatible, 1 case was Bi direction incompatible, nor any effect of PRCA was observed on incidence of GVHD and CMV infection. In conclusion, PRCA is a major complication of patients receiving ABO-incompatible allo-HSCT, while ABO blood group of O/A in recipient/donor pair may be the major high risk factor for PRCA after ABO mismatched allo-HSCT. PMID- 18426667 TI - [Influence of co-culture ex vivo of CD34+ cells from different two units of cord blood on their homing-related adherent molecules expression]. AB - The study was aimed to explore the influence of co-culture ex vivo of CD34+ cells from two units of cord blood (CB) on the homing-related adherent molecule expression of each other. Mesenchymal stem cells (MSCs) were obtained from human bone marrow. Two units of CB CD34+ cells were co-cultured on 12 Gy gamma-ray irradiated MSC layer. Their adherent molecule expressions were assessed by flow cytometry. The results showed that the purity of the isolated CD34+ cells was (98.25+/-0.93)%. After co-culture on MSC layer for 6 days, the proportion of CD34+ cells of each unit was dropped to (60.4+/-6.32)% and (60.2+/-5.12)% respectively, but there was no significant difference from the control groups. The expressions of CD44, CD62L, CD184 and CD26 on CD34+ cells of each unit remained unaffected. The expression of CD162 was downregulated and CD54 was first increased but then dropped to the level before co-culture. But there was no significant difference between the experimental and control groups. In conclusion, co-culture of CD34+ cells from two units of CB may have no effects on the adherent molecule expressions of each other. PMID- 18426668 TI - [Nonmyeloablative peripheral blood stem cell transplantation for chronic myeloid leukemia in chronic and accelerated phases]. AB - The aim of this study was to investigate the effect of nonmyeloablative peripheral blood stem cell transplantation in treatment of chronic myeloid leukemia in chronic phase (CML-CP) and accelerated phase (CML-AP). 24 patients with CML including 16 in CML-CP and 8 in CML-AP were treated with nonmyeloablative conditioning regimen for peripheral blood stem cell transplantation (PBHSCT). The conditioning regimen included fludarabine (30 mg/m(2)x6 d), busulphan [4 mg/(kg.d)x2 d] and CTX [350 mg/(m2.d)x2 d] combined with or without Ara-C. The donors were HLA-identical (n=20) and 5/6 antigen matched (n=4). The dynamic observation of hematopoietic recovery in all patients was carried out. The results indicated that all the patients were successfully engrafted. The mean time for increase of the number of neutrophils to more than 0.5x10(9)/L and platelet more than 20x10(9)/L were 13 days and 11.5 days respectively. Out of 12 patients, 9 patients showed complete donor chimerism and 3 patients showed mixed chimerism at 30 days after transplantation. At 180 days after transplantation, 18 survival patients showed complete donor chimerism. 18 patients remained alive after a median follow-up length of 24 months (4-48 months). 2 cases died of severe acute GVHD and 1 case died of chronic GVHD, 2 cases died of interstitial pneumonia and 1 case died of relapsed. In conclusions, nonmyeloablative peripheral blood stem cell transplantation is an effective therapeutic method for CML patients in chronic phase and accelerated phase. PMID- 18426669 TI - [Dose-effect relationship of DMSO and Tween 80 influencing the growth and viability of murine bone marrow-derived cells in vitro]. AB - This study was purpose to examine the effect of dimethyl sulfoxide (DMSO) and Tween 80 on the growth and viability of stromal cells (BMSC), colony-forming units for granulocytes and macrophages (CFU-GM) and bone marrow endothelial cell line (BMEC) from murine bone marrow in vitro, and to analyze the concentration effect relationship. The colony yields of colony-forming units fibroblastic (CFU F) and CFU-GM were assessed in the murine bone marrow cell cultures at various concentrations of DMSO or Tween 80 and in the control groups. The MTT assay and trypan blue exclusion were used to determine the cell viability and percentage of survival in BMSC and BMEC cultures with or without either of these organic solvents. The results showed that the colony yields of both CFU-F and CFU-GM were decreased significantly (p<0.05 or <0.01) at the concentrations (v/v final) of 2% DMSO or 0.005%-0.01% Tween 80 respectively, as compared with control. The cell viability and percentage of survival of BMSC and BMEC cultures were significantly reduced (p<0.05 or <0.01) at 0.5%-1.0% DMSO or 0.002%-0.005% Tween 80, as compared with control. With the increase of volume fractions of these solvents, the decreased percentages of corresponding measurements were increased by degrees. It is concluded that when the concentration of DMSO or Tween 80 goes to a certain level in cell culture medium, either of the organic solvents has an inhibitory action or/and cytotoxicity on the growth and viability of BMSCs, CFU GM and BMECs. The growth inhibition and cytotoxic response are more significant at higher concentrations of these solvents. PMID- 18426670 TI - [Function of matrix metalloprotenase-2 by RNA interference]. AB - The study was aimed to reveal the effects of matrix metalloproteinase-2 (MMP-2) on cell proliferation, migration, invasion, angiogenesis and cell cycle. The small interfering RNA (siRNA) of MMP-2 transfected into endothelial cells EAhy926, the transformation efficiency at protein and gene levels was evaluated by using flow cytometry and RT-PCR respectively. MTT method was used to detect the proliferation ability of EAhy926. The migration and invasion abilities of EAhy926 induced by two kinds of factors, type IV collagen (COL IV) and fibronectin (Fn) were assayed with Rose Bengal dying method. The changes of angiogenesis were determined by three-dimension culture. The changes of cell cycle and related gene expression were assayed by using flow cytometry and RT-PCR respectively. The results indicated that the proliferation ability of EAhy926 had no obvious difference between interference or not. The migration ability of EAhy926 induced by two kinds of factors, type IV collagen (COL IV) and fibronectin (Fn) was inhibited after interfered by siRNA for 48 hours, and was stronger inhibition to COL IV than Fn (Fn control: 0.581+/-0.012 vs 0.261+/ 0.002; COL IV control: 0.467+/-0.009 vs 0.110+/-0.010, p<0.01). The invasion test had the similar result as migration test. (Fn vs control: 0.365+/-0.012 vs 0.101+/-0.002; COL IV vs control: 0.317+/-0.009 vs 0.102+/-0.010, p<0.01). The angiogenesis ability of endothelial cells dropped to 58.9% of control after interference with siRNA for 48 hours. In the cell cycle experiments, after RNAi for 48 hours and 72 hours, the cell ratio of G1 rose from [(65.9+/-2.53)%; (63.2+/-1.89)%] to [(83.9+/-2.53)%, (89.2+/-1.24)%]% (p<0.01); the cell ratio of S and G2 dropped from [(32.7+/-1.91)%, (37.1+/-2.65)%] to [(18.1+/-1.49)%, (10.2+/-0.85)%] (p<0.01). The results analyzed by semi-quantitative RT-PCR showed that the expression levels of Rb, cyclin D1 PCNA genes dropped to 35%, 51% and 22% of normal control respectively. It is concluded that the expression of MMP-2 does not distinctly correlate with the proliferation of endothelial cells, but plays a very important role in affecting endothelial cell migration, invasion and angiogenesis, and participates in regulating cell cycle through Rb, cyclinD1 and PCNA. PMID- 18426671 TI - [Isolation, culture and identification of two types of endothelial progenitor cells from human umbilical cord blood]. AB - The aim of this study was to establish the method of isolating and culturing endothelial progenitor cells (EPCs) from human umbilical cord blood. Mononuclear cells (MNCs) from human umbilical cord blood were cultured by using culture system supplemented with endothelial cell-conditioned medium. The obtained two types of cells were purified by picking up colonies, identified by uptake of acetylated low-density lipoprotein (Ac-LDL) and binding to lectin [Ulex European Agglatinin (UEA-1)], and were analyzed for the expression of markers by flow cytometry. The results showed that there were significant differences between two types of cells in proliferation, so they were referred as circulating angiogenic cells (CACs) and high proliferative potential endothelial progenitor cells (HPP EPCs), respectively. They were in accordance with the standards of EPCs, could uptake DiI-Ac-LDL and bind to UEA-1, and expressed the markers of endothelial cells, such as CD31, CD144 and vWF detected by immunocytochemistry. The transcription of CD31, KDR, CD144 and ENOS in both of them could be detected by RT-PCR, but FACS analysis showed significant differences of surface marker expression between them. In conclusion, two types of EPCs are successfully obtained by culturing MNCs isolated from human umbilical cord blood using endothelial cell-conditioned medium. PMID- 18426672 TI - [T cells recognizing EBV-epitopes arose in co-culture of peripheral blood mononuclear cells from EB-infected patients and dendritic cells loaded with LMP2 mixed peptides]. AB - The latent membrane protein 2 (LMP2) is a kind of protein expressed by EBV infected cells. This study was aimed to investigate whether the stimulation of peripheral blood mononuclear cells with peptides induces EBV-specific cytotoxic T lymphocytes (CTL). The peptides were mixture of LMP2 protein and available for people with different HLA types. Peripheral blood sample was collected from a patient with EBV-associated hemophagocytic syndrome. The mononuclear cells were isolated and cultured to obtain dendritic cells (DCs). Immature DCs were pulsed with MIX-LMP2 and added with different maturation-promoting factors. The auto-T lymphocytes were stimulated weekly with the harvested mature DCs loaded with MIX LMP2, and totally for two times. Part of isolated lymphocytes was cultured without any stimulation as control. T-cell receptor (TCR) beta spectratyping was used to analyze the distribution of different T cell subgroups before and after culture. The phenotype of T lymphocytes was determined by flow cytometry. The IFN gamma assay was used to estimate specific cytoxic activity of the cultured T cells. The results showed that the distribution of TCRbeta was changed according to analysis of TCR spectratypes. From the distribution of gene families of TCRbeta, the T lymphocytes were oligoclonal before culture, but shifted to a polyclonal after culture in vitro like the normalization of TCR diversity, suggesting the subgroups of lymphocyte could return to normal. The percentage of CD3+, CD3+CD8+ CD3+ CD45RA- CD45 RO+ on T lymphocytes from freshly isolated mononuclear cells were 70.73%, 42.99%, 27.56% respectively. After being stimulated twice with DC loaded with MIX-LMP2, they further increased to 95.17%, 52.54% and 81.41%. The percentages of CD3-CD56+ NK cells and CD4+CD35+ FOXP3+ regulation T cells seldom changed, from 2.12%, 0.03% to 2.35%, 0.02% respectively. The increase of CD3+CD45RA-CD45RO+ cells obviously indicated that most naive T cells could be activated. ELISA for IFN-gamma showed that when DCs loaded with LMP2 peptide were used as target cells, IFN-gamma level secreted by the T cells stimulated with LMP2 peptide-pulsed DCs was 805+/-16 pg/ml and 1729+/ 49 pg/ml, the IFN-gamma level secreted by T cells stimulated twice with LMP2 peptide-pulsed DCs was 956+/-23 pg/ml and 2325+/-58 pg/ml respectively at effector-target ratios of 10:1 and 10:2. They were both significantly higher than that secreted by T cells without any stimulation (441+/-27 pg/m and 557+/-19 pg/ml) (p<0.05). But DCs unpulsed with LMP2 peptide were used as target cells, there were no significant differences between the T cells stimulated with LMP2 peptide-pulsed DCs and the T cells without stimulation (p>0.05). It is concluded that the antigen specific T cells recognizing EBV epitopes can be obtained by using DCs pulsed with MIX-LMP2 peptide in vitro, meanwhile the distribution of T cell subgroups can be changed and normalized. PMID- 18426673 TI - [CMV pp65 gene modified dendritic cells activate autologous T cells]. AB - Cytomegalovirus (CMV) infection is a dangerous complication in patients with chronic graft versus host disease (cGVHD). CMV-specific immunity depends on the activity of T cells. This study was aimed to investigate the effect of CMV pp65 gene modified dendritic cells (DCs) on activation of autologous T cells. Lentivirus system was utilized to introduce the CMV full-length pp65 gene into mouse DCs; CpG-DNA was used to induce mature DCs; flow cytometry and immunofluorescence were used to determine the expression of antigen and IFNgamma in T lymphocytes. The results showed that the DCs were infected with lentivirus at a multiplicity of infection (MOI) of 50 with optimal infectious efficiency of 30%-40%; mature DCs expressing pp65 gene could stimulate autologous naive T cells to express CD69 specifically; mature DCs expressing PP65 could stimulate autologous CD4+ or CD8+ T cells to produce IFNgamma. It is concluded that CMV pp65-modified and CpG-DNA-induced mature DCs can activate CMV-specific T lymphocytes in vitro. PMID- 18426674 TI - Influences of interferon-alpha on expression of Th cytokines and CCR7 in dendritic cells from patients with chronic myeloid leukemia in vitro. AB - This study was aimed to investigate the influences of interferonalpha (IFN-alpha) on expressions of CCR7, interleukin10 (IL-10) and IL-12p70 in dendritic cells (DCs) from patients with chronic myeloid leukemia (CML). In addition to stem cell factor (SCF), granulocyte-macrophage colony stimulating factor (GM-CSF), tumor necrosis factor-alpha (TNF-alpha) and IL-4, IFN-alpha was added to the serum-free medium of DCs. After culture for 10-14 days, phenotypes and function of CML-DCs were evaluated respectively by flow cytometry and methyl thiazolyl tetrazolium (MTT) assay. Chromosome of DCs was analyzed by displaying G banding assay. The concentrations of IL-10 and IL-12P70 in supernatants were evaluated by enzyme linked immunosorbent assay (ELISA). The results showed that the expressions of CD40, CD83, CD86 and CCR7 and the OD value in allogeneic mixed-lymphocyte reaction (MLR) in group with IFN-alpha (300 U/ml) were twice as high as those in group without IFN-alpha. The percentage of Ph1 positive cells and concentrations of IL-10 and IL-12 P70 were reduced in group with IFN-alpha. It is concluded that the defective phenotypes and functions of CML-DCs can be recruited partly by IFN alpha. The mechanism may lie in the facts that expression of CCR7 and co stimulatory molecules is promoted and the inhibitory effect of IL-10 on CML-DCs is relieved partly through the regulation of IFN-alpha. PMID- 18426675 TI - [Immune tolerance induced by exosomes derived from regulatory dendritic cells of mice]. AB - The study was aimed to explore the roles of exosomes derived from regulatory dendritic cells of mice in the induction of immune tolerance. Immature DC (iDC) from mouse bone marrow cells and regulatory DCs (rDC) were induced by treating iDC with TGF-beta1 and IL-10. The phenotype of regulatory DCs and normal DCs were assayed by flow cytometry. Exosomes from immature DCs (iDex) and regulatory DCs (rDex) were isolated by ultracentrifugation and ultrafiltration. A skin transplantation model was established with the recipients BALB/c mice and the donor C57BL/6 mice. Recipients were divided into PBS control group, iDex group (injection 10 microg iDex of donor C57BL/6 mice via tail vein at days 7 and 3 before skin transplantation), rDex group (injection 10 microg rDex of donor C57BL/6 mice via tail vein at days 7 and 3 before skin transplantation). The capacity of the donor mice and the unrelated allogeneic donor mice to stimulate allogeneic T lymphocyte proliferation was examined by mixed lymphocyte culture (MLR). The results showed that TGF-beta1 and IL-10 could down-regulate the expressions of costimulatory molecules, including CD80, CD86 and CD40. The graft mean survival time (MST) in control group, iDex group and rDex group was 7.8, 10.7 and 18.8 days, respectively. There was significant difference in MST between iDex group and control group (p<0.05), and between rDex group and iDex group (p<0.01). The results of MLR assays indicated donor-specific hyporeactivity especially in rDex group, while the tolerant B/C mice were still immunocompetent to unrelated allogeneic DBA mouse. It is concluded that injection iDex or rDex of donor mice via tail vein before skin transplantation induces immunotolerance, and the effect of rDex is more significant. PMID- 18426676 TI - [Morphology, ultrastructure and function of glycosylation-modified chilled blood platelets]. AB - The glycosylation of platelets may prolong their life-span when being transfused after preservation under 4 degrees C, therefore this study was aimed to investigate the effect of glycosylation on morphology, ultrastructure, function and membrane glycoprotein of platelets. The experiments were divided into 3 groups: group preserved in room temperature (RT group), group preserved in 4 degrees C (4T group) and group UDP-Gal glycosylated and preserved in 4 degrees C (U+4T group). The binding rate of RCA I lectin and expression of platelet surface markers CD62P, CD42b were determined by flow cytometry. Morphology and ultrastructure of platelets were observed by light microscopy, scanning electron microscopy (SEM) and transmission electron microscopy (TEM). Platelets aggregation was detected by aggregometer. The results showed that the binding rate of RCAI in U+4T group significantly higher than that in RT group (p<0.01), no obvious changes was found in ultrastructure of glycosylated platelets, as compared with fresh platelets. Some morphologic changes, such as pseudopodium could be observed in 4T group. The aggregation rate of platelets in U+4T group reached to 50% of RT group. The expression levels of CD42b and CD62P, and the binding rate of annexin V in U+4T group were not significantly different from that in RT group. It is concluded that UDP-Gal can effectively cause galactosylation of platelets, and the platelets modified with UDP-Gal remain normal morphology, ultrastructure and function. PMID- 18426677 TI - [Correlations of jak2v617f mutation with clinical and laboratory findings in primary thrombocythemia patients]. AB - To evaluate the frequency of jak2v617f mutation and analyze its correlation with clinical features of primary thrombocythemia (PT) patients, the mutation was detected by allele specific polymerase chain reaction (AS-PCR), the clinical and laboratory data in 66 PT patients with and without jak2v617f mutation were compared and clinical features of these PT patients were retrospectively analyzed. The results showed that among 66 patients, 27 patients (41%) had the mutation. The patients carrying the mutation displayed higher bone marrow erythropoiesis (26.9%+/-9.4% vs 16.3%+/-8.7%, p<0.05), higher ratio of granulopoiesis/erythropoiesis (2.9+/-1.8 vs 5.2+/-2.9, p<0.05) and higher incidence of microvascular disturbances (29.6% vs 5.1%, p<0.05). However, the age, gender, leukocyte and platelet counts, hemoglobin level, bone granulopoiesis, splenomegaly, history of thrombosis and hemorrhage had no difference between patients with and without the mutation. It is concluded that the frequency of jak2v617f mutation in primary thrombocythemia patients is 41%. The presence of the jak2v617f mutation is associated with a higher bone marrow erythropoiesis in primary thrombocythemia patients at diagnosis. PMID- 18426679 TI - [ABO genotyping of Han population in Beijing]. AB - The aim of this study was to establish a diagnostic method for ABO genotyping and to investigate the distribution of ABO genotype in Beijing Han population so as to understand the distribution characteristics and regularity of ABO genotype. An ABO genotyping method was established by using multiplex-PCR-RFLP and PCR-SSP techniques, and the ABO allele frequency in Beijing Han population was investigated. The results showed that A102, O1 and B allele were more common genes in Beijing Han individuals. And A102 allele was predominant in the phenotype A group in this population. Three O2 alleles were found and no A201 allele was found while gene frequency investigation was performed. No A101A101, A101O2, A102O2, BO2 and O2O2 in this population were discovered. It is concluded that the primary regularity of ABO allele distribution in Beijing Han population is found through this study. It provides basic reference for further study of ABO types. PMID- 18426678 TI - [Analysis on expression and molecular basis for ABO glycosyltransferase with dual specificity]. AB - In order to elucidate the expression and molecular genetic background of ABO gene seven samples with ABO discrepancy further identified as bi-specific ABO gene were studied. All these samples were subjected to phenotyping by monoclonal and polyclonal antisera and were then genotyped by direct DNA sequencing and haplotype-sequencing at the exon 6 and 7 of ABO gene. As a result, six ABO dual specific alleles were identified in Chinese population. An antigen expressed by these B (A) or Cis-AB individuals varied from very low level to the normal level, compared with common A blood group samples. In conclusion, molecular genetic backgrounds of two pairs out of four samples in all samples were the same, however, the ABO expression showed diverse. PMID- 18426680 TI - [HLA-B*15 gene polymorphism of registered bone marrow from donors in north Chinese Han population typed by PCR-SBT]. AB - To investigate the characteristics of the allele distribution of HLA-B*15 gene family in Chinese Han population and to study its influence on the selection of clinical transplantation donor, population of a 815 Han in north China from Shaanxi sub-registry of Chinese National Marrow Donor Project was randomly selected and out of them 206 HLA-B*15 positive samples according to the previous known low-resolution typing results were acquired. HLA-B*15 gene polymorphisms of above-mentioned samples and other 17 individuals were analyzed for the first time by polymerase chain reaction sequence-based typing (PCR-SBT) at high-resolution level. The structure differentiation of all HLA-B*15 alleles were analyzed by HLA three-dimensional structure modeling and software Swiss-PdbViewer. The results showed that the distribution of HLA-A, -B, -DRB1 gene of randomly selected 815 samples accorded with Hardy-Weinberg equilibrium and the gene frequency of HLA B*15 was 0.1379. There were a total of 16 kinds of alleles of HLA-B*15 gene family to be obtained, which belonged to 7 kinds of serologic specificities. HLA B*1501, B*1511, B*1502 and B*1518 were the major alleles with a frequency of 0.0485, 0.0215, 0.0178 and 0.0160 respectively, and the constituent ratio of their accumulated frequencies was 75.11%. The each frequency of the other 12 kinds of B*15 alleles was lower than 0.0100. Among the homozygote of 10 samples at low/medial-resolution level, there were only 4 samples to be pur sang homozygote of HLA-B*15xx, --at high-resolution level, and all the homozygote were constituted by respective dominating alleles. HLA three-dimensional structure modeling demonstrated that within the same specificity, gentle structure differentiation not only existed, such as B*1501, 1505, 1507, 1525, 1527, 1532 (each RMSD5) in 12. Apoptotic cell death correlated significantly with viremia because it was demonstrated by higher HCV RNA levels in cases with a high number of apoptotic cells (odds ratio, 2.96; 95% confidence interval, 1.0-8.5; P = .04). No correlation was found between the apoptotic index and hepatitis necroinflammatory activity, CD8 cell count, fibrosis stage, immunosuppressive therapy, or genotype. In HCV-infected renal transplant recipients, apoptotic cell death seems to be associated with high viral load, thus providing indications of viral interference in the pathogenetic process. PMID- 18426735 TI - Diagnosis of hydatidiform moles using p57 immunohistochemistry and HER2 fluorescent in situ hybridization. AB - Fluorescent in situ hybridization (FISH) for HER2 performed on paraffin-embedded tissue samples and immunohistochemical analysis for p57 may be useful ancillary studies to aid in the diagnosis and classification of hydatidiform moles (HMs). HER2 FISH was validated against 24 paraffin-embedded sections of HMs and hydropic abortions and showed an 85% concordance rate. A morphologic assessment based on 44 cases showed 25% disagreement between original and consensus diagnosis based on H&E-stained slides, all of which involved the differential diagnosis of partial mole and hydropic abortion. Immunohistochemical analysis for p57 and HER2 FISH were performed, and a final diagnosis was assigned by using the results from all ancillary studies. p57 staining was absent in 11 of 13 complete moles, and HER2 was triploid in 8 of 10 partial moles .HER2 FISH and immunohistochemical analysis for p57 are useful ancillary techniques in the evaluation of HM, especially when triploid content is seen. PMID- 18426736 TI - Diagnostic role of circulating free plasma DNA detection in patients with localized prostate cancer. AB - To analyze the potential diagnostic relevance of free plasma DNA (FPDNA), we enrolled 64 patients with localized prostate cancer (CaP). FPDNA was quantified by real-time polymerase chain reaction assessment of the HTERT gene in blood samples from 64 patients with CaP and 45 healthy males. Methylation of the GSTP1 gene was used to confirm the neoplastic origin of FPDNA in selected cases. The mean +/- SD levels of FPDNA were higher in patients with CaP (15.4 +/- 10.9 ng/mL) than in control subjects (5.5 +/- 3.5 ng/mL; P <.001). By using the best cutoff value, the sensitivity of the test was 80%, the specificity was 82%, the area under the receiver operating characteristic curve, 0.881. High FPDNA values were significantly associated with pathologic T3 stage (P = . 035). Methylation of the GSTP1 gene was found in 4 (25%) of 16 FPDNA samples and 15 (94%) of 16 tissue samples. Quantification of FPDNA discriminates between patients with CaP and healthy subjects and correlates with pathologic tumor stage. FPDNA is a candidate biomarker for early diagnosis and monitoring of CaP. PMID- 18426737 TI - Assessment of the cost of fine-needle aspiration cytology as a diagnostic tool in patients with thyroid nodules. AB - Fine-needle aspiration cytology (FNAC) is the reference diagnostic tool in patients with thyroid nodules. Because the true diagnosis is based on histopathologic findings, the final diagnosis of nodules not operated on is postponed, impacting the cost. We aimed to determine the cost of FNAC by taking account of diagnostic mistakes, failures, and follow-up of patients who did not have surgery.A Markov model described the management of patients according to initial cytopathologic results. Estimates for accuracy values and follow-up were derived from a retrospective study of 624 patients. Costs were computed from the hospital perspective. Sensitivity analyses were performed. Of the lesions, 381 were cytopathologically classified as benign, 15 as malignant, 57 as "suspicious," and 171 as unsatisfactory. The cost of diagnosis was 1,145 euros . Cost was significantly affected by the unsatisfactory specimen percentage (27.4%), without which the cost would be reduced by 35.5%, to euros 738. Cost depends on cytopathologist performance and the unsatisfactory rate. In the future, routine ultrasound guidance and on-site assessment of cytopathologic adequacy would help reduce costs. PMID- 18426738 TI - Macrophage migration inhibitory factor may be used as an early diagnostic marker in colorectal carcinomas. AB - Recent genetic studies have identified many differentially expressed genes in colorectal carcinomas. For validation of up-regulated genes in colorectal carcinomas, we performed an enzyme-linked immunosorbent assay. Candidate markers were selected from gene expression data for 40 colorectal cancers and 35 matched normal mucosal samples. Based on intensive filtering, 9 genes were selected for the further evaluations. Among them, macrophage migration inhibitory factor (MIF), inhibin betaA, and chemokine ligand 10 were screened, and the results were compared with carcinoembryonic antigen (CEA) in serum samples of 129 patients with colon cancer and 53 healthy control subjects. We found that the serum MIF level was significantly increased in patients with colorectal cancer. Compared with CEA, MIF was more sensitive in early cancer detection (47.3% vs 29.5%). However, the specificity was not as high as that of CEA (90.6% vs 100.0%). Our findings indicate that MIF may be used as a diagnostic marker in colorectal carcinomas. PMID- 18426739 TI - Equivalence of laser scanning cytometric and flow cytometric immunophenotyping of lymphoid lesions in cytologic samples. AB - The immunophenotype of lymphoid cytologic samples obtained by laser scanning cytometry (LSC) and flow cytometry (FC) was compared in 72 cases composed of a series of 23 cases with simultaneous LSC and FC immunophenotyping and a second series of 49 cases in which nonsimultaneous immunophenotyping was performed. In both series, no discordance in the population immunophenotype was found that would result in changes in diagnostic classification, although minor discordance in some antigens was found, predominantly affecting FMC7, CD11c, and CD23. The immunophenotype obtained by LSC shows a high degree of concordance with that obtained by FC and generates results that are diagnostically equivalent. Potential explanations for the discordant markers include differences related to the techniques, differences in the fluorochrome-labeled antibodies, technical factors, differences in antigen expression related to anatomic sites, temporal variations, and interpretive variances. PMID- 18426740 TI - A rapid point-of-care cardiac marker testing strategy facilitates the rapid diagnosis and management of chest pain patients in the emergency department. AB - We compared a rapid, point-of-care multimarker protocol with a single and serial troponin I (TnI)-only protocol in 5,244 patients admitted to the emergency department with chest pain. The diagnosis of acute myocardial infarction (AMI) was based on a doubling myoglobin level accompanied by at least a 50% increase in the creatine kinase (CK)-MB level with no detectable TnI; a doubling of myoglobin level together with any detectable TnI; or a TnI level of 0.4 ng/mL (0.4 microg/L) or more, irrespective of myoglobin or CK-MB results. By using these new criteria, 145 of 148 cases were positive for AMI (positive predictive value [PPV], 92.4%) and 3 were negative, which were also negative by the core laboratory TnI assay. Twelve confirmed non-AMI cases were positive by the new protocol, with 10 of 12 confirmed by the core laboratory as positive for TnI. The negative predictive value (NPV) was 99.9% the overall diagnostic accuracy was 99.7%. The TnI-only protocol had a sensitivity of 68.2% with an NPV of 99.1%. With lower TnI-only cutoffs, 4 patients had false-negative results, and a PPV of 36.4% was observed. Our rapid multimarker protocol seems superior to a TnI-only approach for rapidly triaging patients with chest pain or AMI. PMID- 18426741 TI - Implementation of point-of-care rapid urine testing for drugs of abuse in the emergency department of an academic medical center: impact on test utilization and ED length of stay. AB - We evaluated the impact of implementing a point-of-care (POC) rapid urine test panel for drugs of abuse on turnaround time (TAT), emergency department length of stay (LOS), and laboratory test utilization patterns. The mean TAT from sample collection to results reporting decreased by 69.4%, from 108 to 33 minutes, the mean LOS from 11.1 to 8.1 hours (27% P < .0001), and the median LOS from 8.0 to 7.0 hours (13% P = .0017). A method crossover between the POC and central laboratory methods revealed differences in sensitivity and specificity. Overall, there was no clear preference for either method. Differences in performance for individual drug classes were reconciled by providing interpretive comments with POC results. Following implementation, use of urine testing for drugs of abuse increased by 30%, which was offset by fewer requests for extended toxicology testing in the central laboratory and more selective ordering of toxicology tests not on the POC panel (alcohols and analgesics). The implementation of a POC urine test panel for drugs of abuse decreased LOS and TAT and essentially replaced central laboratory testing for drugs of abuse. Differences in sensitivity and specificity between POC and central laboratory results require provision of interpretive comments with results. PMID- 18426742 TI - LDL species heterogeneity in the atherogenic dyslipidemia of polycystic ovary syndrome. AB - One of the important risk factors for coronary heart disease is dyslipidemia. Several lipid abnormalities have been studied in patients with polycystic ovary syndrome (PCOS), but the relationship between PCOS and low-density lipoprotein (LDL) subclass pattern is not clear. A case-control study was designed to look into lipid differences, and LDL size was analyzed by a newly developed polyacrylamide tube gel electrophoresis method. Results indicated that only PCOS status and serum triglyceride levels were independently associated with LDL particle size. The apolipoprotein (Apo)A-I level was higher in PCOS patients with small dense LDL (sdLDL). PCOS seems to result in smaller LDL particle size and higher ApoA-I levels independent of triglyceride levels. After adjusting for triglyceride levels, other traits of insulin resistance syndrome (IRS) were not associated with LDL size phenotype, suggesting that the IRS-related sdLDL is linked most strongly to alterations in triglyceride levels. PMID- 18426743 TI - The effect of elevated fetal hemoglobin on hemoglobin A1c results: five common hemoglobin A1c methods compared with the IFCC reference method. AB - Hemoglobin A1c (HbA1c) is an important indicator of risk for complications in patients with diabetes mellitus. Elevated fetal hemoglobin (HbF) levels have been reported to interfere with results of some HbA1c methods, but it has generally been assumed that HbA1c results from boronate-affinity methods are not affected by elevated HbF levels. None of the previous studies used the International Federation of Clinical Chemistry and Laboratory Medicine (IFCC) reference method as the comparative HbA1c method. We, therefore, measured HbA1c in samples with normal and elevated HbF levels by several common assay methods and compared the results with those of the IFCC reference method.HbF levels of more than 20% artificially lowered HbA1c results from the Primus CLC 330/385 (Primus Diagnostics, Kansas City, MO), Siemens DCA2000 (Siemens Healthcare Diagnostics, Tarrytown, NY), and Tosoh 2.2+ (Tosoh Bioscience, South San Francisco, CA), but not the Bio-Rad Variant II (Bio-Rad Laboratories, Hercules, CA) and Tosoh G7. Physicians and laboratory professionals need to be aware of potential interference from elevated HbF levels that could affect HbA1c results, including those from boronate-affinity methods. PMID- 18426744 TI - CRP and adiponectin and its oligomers in the metabolic syndrome: evaluation of new laboratory-based biomarkers. AB - The metabolic syndrome (MetS) confers an increased risk for diabetes and cardiovascular disease. Although high-sensitive C-reactive protein (hsCRP) concentrations are higher and adiponectin concentrations lower in MetS, there is no reliable biochemical measure that can capture its various features. We evaluated whether hsCRP, adiponectin, or the ratio of adiponectin or its oligomers, especially the high-molecular-weight (HMW) oligomer, to hsCRP predict MetS in 123 subjects with MetS compared with that in 91 healthy control subjects. MetS subjects had significantly higher hsCRP levels and lower total adiponectin and oligomer levels relative to control subjects (P < .0001). The HMW/total adiponectin and adiponectin/CRP ratios were significantly lower in MetS subjects than control subjects (P < .005). The odds ratio (OR) of MetS using the 75th percentile cutoff for CRP was 3.8 (95% confidence interval [CI], 2.1-6.8) and equivalent to low total adiponectin (OR, 2.5; 95% CI, 1.3-4.5), its oligomers, or the adiponectin/ hsCRP ratio (OR, 2.6; 95% CI, 1.5, 4.8). Thus, measurements of CRP, adiponectin, or its oligomers provide robust biomarkers for predicting MetS. PMID- 18426745 TI - Correlating Bayesian date estimates with climatic events and domestication using a bovine case study. AB - The tribe Bovini contains a number of commercially and culturally important species, such as cattle. Understanding their evolutionary time scale is important for distinguishing between post-glacial and domestication-associated population expansions, but estimates of bovine divergence times have been hindered by a lack of reliable calibration points. We present a Bayesian phylogenetic analysis of 481 mitochondrial D-loop sequences, including 228 radiocarbon-dated ancient DNA sequences, using a multi-demographic coalescent model. By employing the radiocarbon dates as internal calibrations, we co-estimate the bovine phylogeny and divergence times in a relaxed-clock framework. The analysis yields evidence for significant population expansions in both taurine and zebu cattle, European aurochs and yak clades. The divergence age estimates support domestication associated expansion times (less than 12 kyr) for the major haplogroups of cattle. We compare the molecular and palaeontological estimates for the Bison-Bos divergence. PMID- 18426747 TI - Plant community composition, not diversity, regulates soil respiration in grasslands. AB - Soil respiration is responsible for recycling considerable quantities of carbon from terrestrial ecosystems to the atmosphere. There is a growing body of evidence that suggests that the richness of plants in a community can have significant impacts on ecosystem functioning, but the specific influences of plant species richness (SR), plant functional-type richness and plant community composition on soil respiration rates are unknown. Here we use 10-year-old model plant communities, comprising mature plants transplanted into natural non-sterile soil, to determine how the diversity and composition of plant communities influence soil respiration rates. Our analysis revealed that soil respiration was driven by plant community composition and that there was no significant effect of biodiversity at the three levels tested (SR, functional group and species per functional group). Above-ground plant biomass and root density were included in the analysis as covariates and found to have no effect on soil respiration. This finding is important, because it suggests that loss of particular species will have the greatest impact on soil respiration, rather than changes in biodiversity per se. PMID- 18426746 TI - The carotid rete and artiodactyl success. AB - Since the Eocene, the diversity of artiodactyls has increased while that of perissodactyls has decreased. Reasons given for this contrasting pattern are that the evolution of a ruminant digestive tract and improved locomotion in artiodactyls were adaptively advantageous in the highly seasonal post-Eocene climate. We suggest that evolution of a carotid rete, a structure highly developed in artiodactyls but absent in perissodactyls, was at least as important. The rete confers an ability to regulate brain temperature independently of body temperature. The net effect is that in hot ambient conditions artiodactyls are able to conserve energy and water, and in cold ambient conditions they are able to conserve body temperature. In perissodactyls, brain and body temperature change in parallel and thermoregulation requires abundant food and water to warm/cool the body. Consequently, perissodactyls occupy habitats of low seasonality and rich in food and water, such as tropical forests. Conversely, the increased thermoregulatory flexibility of artiodactyls has facilitated invasion of new adaptive zones ranging from the Arctic Circle to deserts and tropical savannahs. PMID- 18426748 TI - Extreme heterochiasmy and nascent sex chromosomes in European tree frogs. AB - We investigated sex-specific recombination rates in Hyla arborea, a species with nascent sex chromosomes and male heterogamety. Twenty microsatellites were clustered into six linkage groups, all showing suppressed or very low recombination in males. Seven markers were sex linked, none of them showing any sign of recombination in males (r=0.00 versus 0.43 on average in females). This opposes classical models of sex chromosome evolution, which envision an initially small differential segment that progressively expands as structural changes accumulate on the Y chromosome. For autosomes, maps were more than 14 times longer in females than in males, which seems the highest ratio documented so far in vertebrates. These results support the pleiotropic model of Haldane and Huxley, according to which recombination is reduced in the heterogametic sex by general modifiers that affect recombination on the whole genome. PMID- 18426749 TI - Tomographic techniques for the study of exceptionally preserved fossils. AB - Three-dimensional fossils, especially those preserving soft-part anatomy, are a rich source of palaeontological information; they can, however, be difficult to work with. Imaging of serial planes through an object (tomography) allows study of both the inside and outside of three-dimensional fossils. Tomography may be performed using physical grinding or sawing coupled with photography, through optical techniques of serial focusing, or using a variety of scanning technologies such as neutron tomography, magnetic resonance imaging and most usefully X-ray computed tomography. This latter technique is applicable at a variety of scales, and when combined with a synchrotron X-ray source can produce very high-quality data that may be augmented by phase-contrast information to enhance contrast. Tomographic data can be visualized in several ways, the most effective of which is the production of isosurface-based 'virtual fossils' that can be manipulated and dissected interactively. PMID- 18426750 TI - Genetic variation in threshold reaction norms for alternative reproductive tactics in male Atlantic salmon, Salmo salar. AB - Alternative reproductive tactics may be a product of adaptive phenotypic plasticity, such that discontinuous variation in life history depends on both the genotype and the environment. Phenotypes that fall below a genetically determined threshold adopt one tactic, while those exceeding the threshold adopt the alternative tactic. We report evidence of genetic variability in maturation thresholds for male Atlantic salmon (Salmo salar) that mature either as large (more than 1 kg) anadromous males or as small (10-150 g) parr. Using a common garden experimental protocol, we find that the growth rate at which the sneaker parr phenotype is expressed differs among pure- and mixed-population crosses. Maturation thresholds of hybrids were intermediate to those of pure crosses, consistent with the hypothesis that the life-history switch points are heritable. Our work provides evidence, for a vertebrate, that thresholds for alternative reproductive tactics differ genetically among populations and can be modelled as discontinuous reaction norms for age and size at maturity. PMID- 18426751 TI - The costs of risky male behaviour: sex differences in seasonal survival in a small sexually monomorphic primate. AB - Male excess mortality is widespread among mammals and frequently interpreted as a cost of sexually selected traits that enhance male reproductive success. Sex differences in the propensity to engage in risky behaviours are often invoked to explain the sex gap in survival. Here, we aim to isolate and quantify the survival consequences of two potentially risky male behavioural strategies in a small sexually monomorphic primate, the grey mouse lemur Microcebus murinus: (i) most females hibernate during a large part of the austral winter, whereas most males remain active and (ii) during the brief annual mating season males roam widely in search of receptive females. Using a 10-year capture-mark-recapture dataset from a population of M. murinus in Kirindy Forest, western Madagascar, we statistically modelled sex-specific seasonal survival probabilities. Surprisingly, we did not find any evidence for direct survival benefits of hibernation-winter survival did not differ between males and females. By contrast, during the breeding season males survived less well than females (sex gap: 16%). Consistent with the 'risky male behaviour' hypothesis, the period for lowered male survival was restricted to the short mating season. Thus, sex differences in survival in a promiscuous mammal can be substantial even in the absence of sexual dimorphism. PMID- 18426752 TI - Progressive island colonization and ancient origin of Hawaiian Metrosideros (Myrtaceae). AB - Knowledge of the evolutionary history of plants that are ecologically dominant in modern ecosystems is critical to understanding the historical development of those ecosystems. Metrosideros is a plant genus found in many ecological and altitudinal zones throughout the Pacific. In the Hawaiian Islands, Metrosideros polymorpha is an ecologically dominant species and is also highly polymorphic in both growth form and ecology. Using 10 non-coding chloroplast regions, we investigated haplotype diversity in the five currently recognized Hawaiian Metrosideros species and an established out-group, Metrosideros collina, from French Polynesia. Multiple haplotype groups were found, but these did not match morphological delimitations. Alternative morphologies sharing the same haplotype, as well as similar morphologies occurring within several distinct island clades, could be the result of developmental plasticity, parallel evolution or chloroplast capture. The geographical structure of the data is consistent with a pattern of age progressive island colonizations and suggests de novo intra-island diversification. If single colonization events resulted in a similar array of morphologies on each island, this would represent parallel radiations within a single, highly polymorphic species. However, we were unable to resolve whether the pattern is instead explained by ancient introgression and incomplete lineage sorting resulting in repeated chloroplast capture. Using several calibration methods, we estimate the colonization of the Hawaiian Islands to be potentially as old as 3.9 (-6.3) Myr with an ancestral position for Kaua'i in the colonization and evolution of Metrosideros in the Hawaiian Islands. This would represent a more ancient arrival of Metrosideros to this region than previous studies have suggested. PMID- 18426753 TI - Mimetic butterflies support Wallace's model of sexual dimorphism. AB - Theoretical and empirical observations generally support Darwin's view that sexual dimorphism evolves due to sexual selection on, and deviation in, exaggerated male traits. Wallace presented a radical alternative, which is largely untested, that sexual dimorphism results from naturally selected deviation in protective female coloration. This leads to the prediction that deviation in female rather than male phenotype causes sexual dimorphism. Here I test Wallace's model of sexual dimorphism by tracing the evolutionary history of Batesian mimicry-an example of naturally selected protective coloration-on a molecular phylogeny of Papilio butterflies. I show that sexual dimorphism in Papilio is significantly correlated with both female-limited Batesian mimicry, where females are mimetic and males are non-mimetic, and with the deviation of female wing colour patterns from the ancestral patterns conserved in males. Thus, Wallace's model largely explains sexual dimorphism in Papilio. This finding, along with indirect support from recent studies on birds and lizards, suggests that Wallace's model may be more widely useful in explaining sexual dimorphism. These results also highlight the contribution of naturally selected female traits in driving phenotypic divergence between species, instead of merely facilitating the divergence in male sexual traits as described by Darwin's model. PMID- 18426754 TI - Cone visual pigments in two marsupial species: the fat-tailed dunnart (Sminthopsis crassicaudata) and the honey possum (Tarsipes rostratus). AB - Uniquely for non-primate mammals, three classes of cone photoreceptors have been previously identified by microspectrophotometry in two marsupial species: the polyprotodont fat-tailed dunnart (Sminthopsis crassicaudata) and the diprotodont honey possum (Tarsipes rostratus). This report focuses on the genetic basis for these three pigments. Two cone pigments were amplified from retinal cDNA of both species and identified by phylogenetics as members of the short wavelength sensitive 1 (SWS1) and long wavelength-sensitive (LWS) opsin classes. In vitro expression of the two sequences from the fat-tailed dunnart confirmed the peak absorbances at 363 nm in the UV for the SWS1 pigment and 533 nm for the LWS pigment. No additional expressed cone opsin sequences that could account for the middle wavelength cones could be amplified. However, amplification from the fat tailed dunnart genomic DNA with RH1 (rod) opsin primer pairs identified two genes with identical coding regions but sequence differences in introns 2 and 3. Uniquely therefore for a mammal, the fat-tailed dunnart has two copies of an RH1 opsin gene. This raises the possibility that the middle wavelength cones express a rod rather than a cone pigment. PMID- 18426755 TI - Perceptual uncertainty and line-call challenges in professional tennis. AB - Fast-moving sports such as tennis require both players and match officials to make rapid accurate perceptual decisions about dynamic events in the visual world. Disagreements arise regularly, leading to disputes about decisions such as line calls. A number of factors must contribute to these disputes, including lapses in concentration, bias and gamesmanship. Fundamental uncertainty or variability in the sensory information supporting decisions must also play a role. Modern technological innovations now provide detailed and accurate physical information that can be compared against the decisions of players and officials. The present paper uses this psychophysical data to assess the significance of perceptual limitations as a contributor to real-world decisions in professional tennis. A detailed analysis is presented of a large body of data on line-call challenges in professional tennis tournaments over the last 2 years. Results reveal that the vast majority of challenges can be explained in a direct highly predictable manner by a simple model of uncertainty in perceptual information processing. Both players and line judges are remarkably accurate at judging ball bounce position, with a positional uncertainty of less than 40mm. Line judges are more reliable than players. Judgements are more difficult for balls bouncing near base and service lines than those bouncing near side and centre lines. There is no evidence for significant errors in localization due to image motion. PMID- 18426757 TI - The neuroscience of primate intellectual evolution: natural selection and passive and intentional niche construction. AB - We trained Japanese macaque monkeys to use tools, an advanced cognitive function monkeys do not exhibit in the wild, and then examined their brains for signs of modification. Following tool-use training, we observed neurophysiological, molecular genetic and morphological changes within the monkey brain. Despite being 'artificially' induced, these novel behaviours and neural connectivity patterns reveal overlap with those of humans. Thus, they may provide us with a novel experimental platform for studying the mechanisms of human intelligence, for revealing the evolutionary path that created these mechanisms from the 'raw material' of the non-human primate brain, and for deepening our understanding of what cognitive abilities are and of those that are not uniquely human. On these bases, we propose a theory of 'intentional niche construction' as an extension of natural selection in order to reveal the evolutionary mechanisms that forged the uniquely intelligent human brain. PMID- 18426758 TI - [Endproducts and receptors of advanced glycation and lipoxidation (AGE, ALE, RAGE) and chronic diseases from the perspective of food and nutrition]. AB - BACKGROUND: Chronic diseases as well as complications to acute and chronic disease are repeatedly associated with accumulation in the body of glycated and lipoxidated proteins and peptides. These molecules are strongly associated with activation of a specific receptor called RAGE and a long-lasting exaggerated level of inflammation in the body. METHODS: PubMed reports in excess of 5000 papers plus about 14000 articles about the related HbA 1c , most of them published in the last five years. Most of available abstracts have been read and circa 800 full papers studied in detail. RESULTS: RAGE, a member of the immunoglobulin superfamily of cell surface molecules and receptor for advanced glycation endproducts, functions as a master switch, induces sustained activation of NF-kappaB, suppresses a series of endogenous autoregulatory functions and converts long-lasting pro-inflammatory signals into sustained cellular dysfunction and disease. Its activation is associated with high levels of dysfunctioning proteins in body fluids and tissues, and strongly associated with a series of diseases from allergy and Alzheimer to rheumatoid arthritis and urogenital disorders. Heat-treatment, irradiation and ionisation of foods increase the content in foods of AGE/ALE. CONCLUSIONS: Some processed foods are much like tobacco smoking great contributors to accumulation of glycated and lipoxidated molecules in the tissues. Change of life style: avoidance of foods rich in deranged proteins and peptides and increased consumption of antioxidants, especially polyphenols counteracts such a development. PMID- 18426756 TI - Progress in the pathogenesis and genetics of Parkinson's disease. AB - Recent progresses in the pathogenesis of sporadic Parkinson's disease (PD) and genetics of familial PD are reviewed. There are common molecular events between sporadic and familial PD, particularly between sporadic PD and PARK1-linked PD due to alpha-synuclein (SNCA) mutations. In sporadic form, interaction of genetic predisposition and environmental factors is probably a primary event inducing mitochondrial dysfunction and oxidative damage resulting in oligomer and aggregate formations of alpha-synuclein. In PARK1-linked PD, mutant alpha synuclein proteins initiate the disease process as they have increased tendency for self-aggregation. As highly phosphorylated aggregated proteins are deposited in nigral neurons in PD, dysfunctions of proteolytic systems, i.e. the ubiquitin proteasome system and autophagy-lysosomal pathway, seem to be contributing to the final neurodegenerative process. Studies on the molecular mechanisms of nigral neuronal death in familial forms of PD will contribute further on the understanding of the pathogenesis of sporadic PD. PMID- 18426759 TI - [Evaluation of liver function before living donor liver transplantation and liver resection]. AB - Liver resection is the curative therapeutic option for hepatocellular carcinoma, biliary tumors, metastases of colorectal and other extrahepatic tumors, living donor liver transplantation and other benign liver diseases. AIM OF STUDY: To summarize the evaluation methods of liver function before living donor liver transplantation and liver resection. METHOD: We summarize the literature about the evaluation of liver function. RESULTS: Perioperative mortality is determined mostly by the extent of preoperative evaluation focused on the liver. After resection the remnant liver parenchyma must cope with the challenge caused by increased metabolism, portal overflow, decreased vascular bed and biliary tract and oxidative stress following the operation. If the remnant liver is unable to grow up to this challenge, acute liver failure occurs. This maintains the necessity of determining the hepatic functional reserve and the hepatic remnant volume. Child-Pugh classification is widely spread to predict outcome. Dynamic functional tests such as indocyanine green retention test, galactosyl human serum albumin scintigraphy and aminopyrine breath tests can be used to evaluate hepatic reserve. To determine remnant liver volume modern imaging processes such as CT volumetry and hepatobiliary scintigraphy are available. CONCLUSION: After the detailed evaluation resection can be limited to an extent which is oncologically radical enough (1% remnant liver tissue/kg) and spares parenchyma which can ensure survival yet. With careful preoperative examination mortality can be reduced even to reach zero. PMID- 18426760 TI - [Old age and the kidneys]. AB - Age-related changes in renal morphology and function cannot be regarded physiological. The number of glomeruli falls, sclerotic glomeruli and aglomerular arterioles develop. Besides tubular atrophy interstitial fibrosis is often seen, and the age-related vascular changes strongly affect the kidneys. Renal blood flow and GFR decrease, without concomitant changes in se-creatinine. Disorders of tubular transport manifest mainly in salt- and water-excretion and lead to hyposthenuria. The pathogenesis of these age-related changes is not fully understood. Nevertheless, such changes impair the excretory functions and the pharmacokinetics of drugs. In real chronic renal failure other functions (erythropoietin production, vitamin-D, Ca and P metabolism) are also impaired. Due to more frequent occurrence of systemic diseases (diabetes, hypertension, etc.) in the elderly, real chronic renal failure is also more common, and various forms of acute renal failure develop more easily. PMID- 18426761 TI - [The first case of human alveolar Echinococcosis in Hungary]. AB - Infection caused by Echinococcus multilocularis is a rare helminthiasis, human cases have not been diagnosed in Hungary until now. The endemic region is Central Europe; the occurrence of this infection has been reported from most of the neighbouring countries; however, E. multilocularis has been found in the red fox population in Hungary. Summarizing the recent knowledge concerning epidemiological, clinical patterns and therapeutic options, the authors describe the first Hungarian case of alveolar echinococcosis. In the presence of appropriate clinical findings, the possibility of this rare infection has to be considered in the differential diagnosis of infiltrative hepatic lesions. PMID- 18426762 TI - [Long-term survival after nasal NK/T cell lymphoma]. AB - The nasal NK/T cell lymphoma is a rare, extranodal non-Hodgkin lymphoma in western civilizations, which has poor prognosis. The Epstein-Barr virus can be detected in tumor cells in nearly all cases. There are no definite treatment guidelines in our days. There is no significant difference in survival between radiotherapy and chemotherapy according to Asian studies. In this case study we show our diagnostic procedures, our treatment options and we present the summary of this illness based on the data found in the literature. PMID- 18426763 TI - [Chronic lymphocytic leukemia]. PMID- 18426768 TI - Novel preparation of graded porous structures for medical engineering. AB - The gradation of porosity in a biomaterial can be very useful for a variety of medical engineering applications such as filtration, bone replacement and implant development. However, the preparation of such structures is not a technologically trivial task and replication methods do not offer an easy solution. In this work, we elucidate the preparation of structures having a graded porosity by electrohydrodynamic spraying, using zirconia (ZrO2), which is widely used in biomedical and other applications. The processes are generic and can be achieved using other bioactive ceramics with similar particle characteristics. The pores on the sprayed surface, the innermost surface and lengthwise cross sections have been analysed in addition to the change in depth of penetration as a function of spraying time. Control of porosity, pore size and depth of penetration has been obtained by varying parameters such as the spraying time, sintering temperature and the sacrificial template. It has been possible to obtain structures with interconnected pore networks of pore size greater than 100microm as well as scattered pores smaller than 10microm in size. PMID- 18426769 TI - Current techniques for single-cell lysis. AB - Owing to the small quantities of analytes and small volumes involved in single cell analysis techniques, manipulation strategies must be chosen carefully. The lysis of single cells for downstream chemical analysis in capillaries and lab-on a-chip devices can be achieved by optical, acoustic, mechanical, electrical or chemical means, each having their respective strengths and weaknesses. Selection of the most appropriate lysis method will depend on the particulars of the downstream cell lysate processing. Ultrafast lysis techniques such as the use of highly focused laser pulses or pulses of high voltage are suitable for applications requiring high temporal resolution. Other factors, such as whether the cells are adherent or in suspension and whether the proteins to be collected are desired to be native or denatured, will determine the suitability of detergent-based lysis methods. Therefore, careful selection of the proper lysis technique is essential for gathering accurate data from single cells. PMID- 18426770 TI - Rate constants rather than biochemical mechanism determine behaviour of genetic clocks. AB - Many biological systems contain both positive and negative feedbacks. These are often classified as resonators or integrators. Resonators respond preferentially to oscillating signals of a particular frequency. Integrators, on the other hand, accumulate a response to signals. Computational neuroscientists often refer to neurons showing integrator properties as type I neurons and those showing resonator properties as type II neurons. Guantes & Poyatos have shown that type I or type II behaviour can be seen in genetic clocks. They argue that when negative feedback occurs through transcription regulation and post-translationally, genetic clocks act as integrators and resonators, respectively. Here we show that either behaviour can be seen with either design and in a wide range of genetic clocks. This highlights the importance of parameters rather than biochemical mechanism in determining the system behaviour. PMID- 18426771 TI - Quantitative approaches to the study of bistability in the lac operon of Escherichia coli. AB - In this paper, the history and importance of the lac operon in the development of molecular and systems biology are briefly reviewed. We start by presenting a description of the regulatory mechanisms in this operon, taking into account the most recent discoveries. Then we offer a survey of the history of the lac operon, including the discovery of its main elements and the subsequent influence on the development of molecular and systems biology. Next the bistable behaviour of the operon is discussed, both with respect to its discovery and its molecular origin. A review of the literature in which this bistable phenomenon has been studied from a mathematical modelling viewpoint is then given. We conclude with some brief remarks. PMID- 18426772 TI - How much of protein sequence space has been explored by life on Earth? AB - We suggest that the vastness of protein sequence space is actually completely explorable during the populating of the Earth by life by considering upper and lower limits for the number of organisms, genome size, mutation rate and the number of functionally distinct classes of amino acids. We conclude that rather than life having explored only an infinitesimally small part of sequence space in the last 4 Gyr, it is instead quite plausible for all of functional protein sequence space to have been explored and that furthermore, at the molecular level, there is no role for contingency. PMID- 18426773 TI - Combining experiments and modelling to understand size regulation in Dictyostelium discoideum. AB - Little is known about how the sizes of specific organs and tissues are regulated. To try to understand these mechanisms, we have been using a combination of modelling and experiments to study the simple system Dictyostelium discoideum, which forms approximately 20000 cell groups. We found that cells secrete a factor, and as the number of cells increases, the concentration of the factor increases. Diffusion calculations indicated that this lets cells sense the local cell density. Computer simulations predicted, and experiments then showed, that this factor decreases cell-cell adhesion and increases random cell motility. In a group, adhesion forces keep cells together, while random motility forces cause cells to pull apart and separate from each other. As the group size increases above a threshold, the factor concentration goes above a threshold and the cells switch from an adhered state to a separated state. This causes excessively large groups to break apart and/or dissipate, creating an upper limit to group size. In this review, we focus on how computer simulations made testable predictions that led the way to understanding the size regulation mechanism mediated by this factor. PMID- 18426774 TI - Synchrony and entrainment properties of robust circadian oscillators. AB - Systems theoretic tools (i.e. mathematical modelling, control, and feedback design) advance the understanding of robust performance in complex biological networks. We highlight phase entrainment as a key performance measure used to investigate dynamics of a single deterministic circadian oscillator for the purpose of generating insight into the behaviour of a population of (synchronized) oscillators. More specifically, the analysis of phase characteristics may facilitate the identification of appropriate coupling mechanisms for the ensemble of noisy (stochastic) circadian clocks. Phase also serves as a critical control objective to correct mismatch between the biological clock and its environment. Thus, we introduce methods of investigating synchrony and entrainment in both stochastic and deterministic frameworks, and as a property of a single oscillator or population of coupled oscillators. PMID- 18426775 TI - Testing the hypothesis that amphiphilic antineoplastic lipid analogues act through reduction of membrane curvature elastic stress. AB - The alkyllysophospholipid (ALP) analogues Mitelfosine and Edelfosine are anticancer drugs whose mode of action is still the subject of debate. It is agreed that the primary interaction of these compounds is with cellular membranes. Furthermore, the membrane-associated protein CTP: phosphocholine cytidylyltransferase (CCT) has been proposed as the critical target. We present the evaluation of our hypothesis that ALP analogues disrupt membrane curvature elastic stress and inhibit membrane-associated protein activity (e.g. CCT), ultimately resulting in apoptosis. This hypothesis was tested by evaluating structure-activity relationships of ALPs from the literature. In addition we characterized the lipid typology, cytotoxicity and critical micelle concentration of novel ALP analogues that we synthesized. Overall we find the literature data and our experimental data provide excellent support for the hypothesis, which predicts that the most potent ALP analogues will be type I lipids. PMID- 18426776 TI - Random walk models in biology. AB - Mathematical modelling of the movement of animals, micro-organisms and cells is of great relevance in the fields of biology, ecology and medicine. Movement models can take many different forms, but the most widely used are based on the extensions of simple random walk processes. In this review paper, our aim is twofold: to introduce the mathematics behind random walks in a straightforward manner and to explain how such models can be used to aid our understanding of biological processes. We introduce the mathematical theory behind the simple random walk and explain how this relates to Brownian motion and diffusive processes in general. We demonstrate how these simple models can be extended to include drift and waiting times or be used to calculate first passage times. We discuss biased random walks and show how hyperbolic models can be used to generate correlated random walks. We cover two main applications of the random walk model. Firstly, we review models and results relating to the movement, dispersal and population redistribution of animals and micro-organisms. This includes direct calculation of mean squared displacement, mean dispersal distance, tortuosity measures, as well as possible limitations of these model approaches. Secondly, oriented movement and chemotaxis models are reviewed. General hyperbolic models based on the linear transport equation are introduced and we show how a reinforced random walk can be used to model movement where the individual changes its environment. We discuss the applications of these models in the context of cell migration leading to blood vessel growth (angiogenesis). Finally, we discuss how the various random walk models and approaches are related and the connections that underpin many of the key processes involved. PMID- 18426777 TI - A geometric flow-based approach for diffusion tensor image segmentation. AB - Diffusion tensor magnetic resonance imaging (DT-MRI, shortened as DTI) produces, from a set of diffusion-weighted magnetic resonance images, tensor-valued images where each voxel is assigned a 3x3 symmetric, positive-definite matrix. This tensor is simply the covariance matrix of a local Gaussian process with zero mean, modelling the average motion of water molecules. We propose a three dimensional geometric flow-based model to segment the main core of cerebral white matter fibre tracts from DTI. The segmentation is carried out with a front propagation algorithm. The front is a three-dimensional surface that evolves along its normal direction with speed that is proportional to a linear combination of two measures: a similarity measure and a consistency measure. The similarity measure computes the similarity of the diffusion tensors at a voxel and its neighbouring voxels along the normal to the front; the consistency measure is able to speed up the propagation at locations where the evolving front is more consistent with the diffusion tensor field, to remove noise effect to some extent, and thus to improve results. We validate the proposed model and compare it with some other methods using synthetic and human brain DTI data; experimental results indicate that the proposed model improves the accuracy and efficiency in segmentation. PMID- 18426780 TI - Self-rated health in adolescence: a multifactorial composite. AB - BACKGROUND AND AIMS: Self-rated health is an important health predictor, and it has only rarely been studied in adolescents. This study examined the relationships between self-rated health and a broad spectrum of structural, medical, psychological, and social variables. The association between these variables and negative health rating through to good health rating versus good to very good health rating was also compared. METHODS: Analyses were based on cross sectional data from the Young-HUNT II study in Norway. A total of 2,800 students aged 16 to 20 years participated, with a response rate of 81%. Separate logistic regression analyses for each gender were performed for a broad set of independent variables with self-rated health as the dependent variable. The effect of the variables at the negative (poor/not good) and positive (very good) ends of the scale were estimated and compared. RESULTS: Self-rated health in adolescence was significantly associated with a broad spectrum of independent variables reflecting medical, social, and personal factors. The associations were also present in multivariate analyses controlling for the interrelations between the independent variables. The negative and positive ends of the scale were affected in much the same way. The association with general well-being was especially strong. CONCLUSIONS: Adolescents conceptualize health as a construct related to medical, psychological, social, and lifestyle factors. Positive rating of health was affected in a similar manner to negative rating. However, the absolute importance of hampering positive health may be greater because of the higher prevalence of such health ratings. PMID- 18426778 TI - L-4F treatment reduces adiposity, increases adiponectin levels, and improves insulin sensitivity in obese mice. AB - We hypothesized that the apolipoprotein mimetic peptide L-4F, which induces arterial anti-oxidative enzymes and is vasoprotective in a rat model of diabetes, would ameliorate insulin resistance and diabetes in obese mice. L-4F (2 mg/kg/d) administered to ob/ob mice for 6 weeks limited weight gain without altering food intake, decreased visceral (P < 0.02) and subcutaneous (P < 0.045) fat content, decreased plasma IL-1beta and IL-6 levels (P < 0.05) and increased insulin sensitivity, resulting in decreased glucose (P < 0.001) and insulin (P < 0.036) levels. In addition, L-4F treatment increased aortic and bone marrow heme oxygenase (HO) activity and decreased aortic and bone marrow superoxide production (P < 0.001). L-4F treatment increased serum adiponectin levels (P < 0.037) and decreased adipogenesis in mouse bone marrow (P < 0.039) and in cultures of human bone marrow-derived mesenchymal stem cells (P < 0.022). This was manifested by reduced adiposity, improved insulin sensitivity, improved glucose tolerance, increased plasma adiponectin levels, and reduced IL-1beta and IL-6 levels in obese mice. This study highlights the existence of a temporal relationship between HO-1 and adiponectin that is positively affected by L-4F in the ob/ob mouse model of diabetes, resulting in the amelioration of the deleterious effects of diabetes. PMID- 18426781 TI - Associations between strain in domestic work and self-rated health: a study of employed women in Sweden. AB - AIM: The aim of this study was to analyse the association between strain in domestic work and self-rated health among employed women in Sweden, using two different methods of measuring strain in domestic work. METHODS: Questionnaire data were collected on health and living conditions in paid and unpaid work for employed women (n=1,417), aged 17-64 years. "Domestic job strain'' was an application of the demand-control model developed by Karasek and Theorell, and "Domestic work equity and marital satisfaction'' was measured by questions on the division of and responsibility for domestic work and relationship with spouse/cohabiter. Self-rated health was measured using the SF-36 Health Survey. Associations were analysed by bivariate and multivariate linear regression analyses, and reported as standardized regression coefficients. RESULTS: Higher strain in domestic work was associated with lower self-rated health, also after controlling for potential confounders and according to both strain measures. "Domestic work equity and marital satisfaction'' showed for example negative associations with mental health beta -0.211 (p<0.001), vitality beta -0.195 (p<0.001), social function -0.132 (p<0.01) and physical role beta -0.115 (p<0.01). The highest associations between "Domestic job strain'' and SF-36 were found for vitality beta -0.156 (p<0.001), mental health beta -0.123 (p<0.001). CONCLUSIONS: Strain in domestic work, including perceived inequity in the relationship and lack of a satisfactory relationship with a spouse/cohabiter, was associated with lower self-rated health in this cross-sectional study. Future research needs to address the specific importance of strain in domestic work as a contributory factor to women's ill-health. PMID- 18426782 TI - Social capital, political trust and self rated-health: a population-based study in southern Sweden. AB - AIM: To investigate the association between political trust (an aspect of institutional trust) and self-rated health, taking generalized (horizontal) trust in other people into account. METHODS: The 2004 public health survey in Skane is a cross-sectional postal questionnaire study answered by 27,963 respondents aged 18-80 years, yielding a 59% response rate. A logistic regression model was used to investigate the associations between political trust in the Riksdag (national parliament) and self-rated health. Multivariate analyses of political trust and self-rated health were performed in order to investigate the importance of possible confounders. RESULTS: Poor health was reported by 28.7% of the men and 33.2% of the women. In total, 17.3% and 11.6% of the male and female respondents, respectively, reported that they had no trust at all in the Riksdag. The addition of generalized (horizontal) trust in the multivariate models reduced the odds ratios of poor self-rated health in the "no political trust at all'' category as compared to the "very high political trust'' category from 2.4 (1.8-3.1) to 2.1 (1.6-2.7) among men and from 1.9 (1.4-2.4) to 1.6 (1.3-2.1) among women. CONCLUSIONS: Low political trust in the Riksdag seems to be significantly associated with poor self-rated health, even after adjustments for plausible confounders, including generalized (horizontal) trust. PMID- 18426783 TI - Why older people living with a spouse are less likely to be institutionalized: the role of socioeconomic factors and health characteristics. AB - AIMS: To examine whether the lower risk of institutionalization among older adults living with a spouse as compared with those living alone or with other persons could be explained by socioeconomic factors, housing, and chronic medical conditions. METHODS: We used population-based follow-up data on Finnish adults aged 65 years and over (N=280,722), covering the period from January 1998 to September 2003, to analyse the risk of entering into long-term institutional care by living arrangements. Kaplan-Meier estimates and Cox regression models were applied. RESULTS: Among men, those living alone had a 70% higher risk and those living with other persons a 56% higher risk of being institutionalized than those living with a spouse, independently of age, region, and urbanicity. The corresponding figures for women were 29% and 21%. Among men, the lower risk of institutionalization among those living with a spouse than among those living alone was partly explained by higher educational level, occupation-based social class, household income, home ownership, house type, better housing conditions, and lower likelihood of having depressive symptoms. Almost the same factors helped to explain the lower risk among women, except that those living with a spouse were not advantaged in terms of education or having fewer chronic diseases. CONCLUSIONS: As controlling for socioeconomic factors, housing and health characteristics explained only 35-43% of the lower risk of institutionalization among those living with a spouse as compared to those living alone, having a spouse seems to have a major independent role in preventing and delaying institutionalization among older men and women. PMID- 18426784 TI - Increase in social inequality in health expectancy in Denmark. AB - AIMS: Health expectancy represents the average lifetime in various states of health and differs among social groups. The purpose of the study was to determine trends in social inequality in health expectancy since 1994 between groups with high, medium and low educational levels in Denmark. METHODS: The study was based on data from nationwide registers on educational level and mortality during the period 1994-2005 and data on health status derived from the Danish Health Interview Surveys carried out in 1994, 2000 and 2005. Expected lifetime in self rated good and poor health, lifetime without and with longstanding illness and expected lifetime without and with long-lasting difficulties or restrictions were estimated by Sullivan's method. RESULTS: Between 1994 and 2005, life expectancy at age 30 years increased by 1.9 years for men and 1.5 years for women with a low educational level. For people with a high educational level, the increase was 2.7 years for men and 2.2 years for women. The difference between people with low and high educational level in expected lifetime in self-rated good health increased by 2.0 and 1.3 years for 30-year-old men and women, respectively. The social gap also increased for other indicators. CONCLUSIONS: During the past 12 years, social inequality in life expectancy and health expectancy has increased in Denmark, but the proportion of the population with a low educational level has decreased. PMID- 18426785 TI - Reliability and validity of self-reported physical activity in the Nord-Trondelag Health Study: HUNT 1. AB - BACKGROUND: A large health survey was previously conducted in 1984-86, the Nord Trondelag Health Study (HUNT 1), and another was conducted in 1995-97 (HUNT 2). A third, HUNT 3, started in 2006. However, the physical activity (PA) questionnaires have not yet been validated. AIMS: To assess the reliability and validity of the self-reported physical activity questionnaire in the Nord Trondelag Health Study (HUNT 1). METHODS: The HUNT 1 questionnaire was administered to a random sample of 108 healthy men aged 20-39 years. Repeatability was assessed with a repeat questionnaire after one week, and validity by comparing results with direct measurement of VO(2) during maximal work on a treadmill, with ActiReg, an instrument that measures PA and energy expenditure (EE) and with the International Physical Activity Questionnaire (IPAQ). ActiReg records the main body positions (stand, sit, bent forward and lie) together with the motion of the trunk and/or one leg each second. RESULTS: The results indicated strong, significant agreement on test-retest (weighted kappa frequency, r=0.80, intensity, r=0.82, and duration, r=0.69). We found a moderate, significant correlation, r=0.48 (p< or =0.01), between the index based on questionnaire responses and VO(2max.) Metabolic equivalent (MET) values of 6 or more from ActiReg and "vigorous activity'' from the IPAQ most strongly correlated with the index (r=0.39, r=0.55, respectively). Associations of other measures obtained from ActiReg with questionnaire responses were weaker. CONCLUSIONS: Our results indicate that the PA questionnaire in HUNT 1 is reproducible and provides a useful measure of leisure-time PA for men. The questionnaire is very short, and compared favourably with much longer instruments for assessment of more vigorous PA. It should be an appropriate tool for use in further epidemiological studies, particularly when the interest is in aspects of PA reflected in fitness or METs greater than 6. PMID- 18426787 TI - Guidelines improve general trend of lowered cholesterol levels in type 2 diabetes patients in spite of low adherence. AB - BACKGROUND: New guidelines were introduced in Sweden in 1999 to reduce the risk of cardiovascular disease among diabetes patients. The impact of the new guidelines on lipid levels in a diabetes patient population has not been extensively studied. Our aim was to study whether the introduction of treatment goals for dyslipidaemia was reflected in lower cholesterol levels in patients with diabetes in a general population. METHODS: In a population of 59,338 individuals, 971 (1.6%) had diabetes. All subjects were 40, 50 or 60 years of age and participated in the Vasterbotten Intervention Programme in 1995-2004. Cholesterol levels and use of lipid-lowering drugs were measured, and trends in cholesterol levels were analysed before and after the guidelines were introduced in 1999. RESULTS: In this effectiveness study, there was a marked decrease in mean plasma total cholesterol levels among patients with diabetes (5.79 (+/- 1.21) mmol/1 in 1995-1999 and 5.07 (+/- 1.00) mmol/1 in 2000-2004 (p<0.001)) as well as in the non-diabetic population (5.79 (+/-1.15) mmol/1 and 5.41 (+/-1.07) mmol/1 (p<0.001)). The trend in diabetes patients was influenced by increased use of lipid-lowering agents, even though only 25.3% (male/female 26.8%/23.2%) of the diabetes patients received lipid-lowering treatment after the introduction of the new guidelines. CONCLUSIONS: Since the introduction of the guidelines, an increasing number of diabetes patients are receiving lipid-lowering drugs, which enhance a strong general trend of lowered cholesterol levels. Yet, the vast majority of diabetes patients with hypercholesterolaemia still do not receive medical treatment in accordance with the guidelines. PMID- 18426786 TI - History of and factors associated with diabetic foot ulcers in Norway: the Nord Trondelag Health Study. AB - AIMS: To determine the proportion of people with diabetes mellitus reporting a history of foot ulcer and to investigate factors associated with this adverse outcome. METHODS: All inhabitants aged 20 years and older residing in a large geographic region were invited to participate in the Nord-Trondelag Health Study, 71% (n=65,604) attended. Those reporting diabetes (n=1,972) were invited to take part in an ancillary study on diabetes. Based on 1,494 responses to the question: "Have you had a foot ulcer that required more than three weeks to heal'', the proportion with a history of foot ulcer was estimated. RESULTS: The overall proportion with a history of foot ulcer was 10.4% (95% CI 8.8-11.9%). In the final multivariate logistic regression model, significant factors for a foot ulcer history included age > or =75 years (OR 1.8, 95% CI 1.2-2.8), height (men>175 cm, women>161 cm) (1.9, 95% CI 1.3-2.8), gender (male) (1.5, 95% CI 1.03 2.2), using insulin (1.6, 95% CI 1.1-2.4), and macrovascular complications (1.8, 95% CI 1.2-2.6). CONCLUSIONS: The proportion of people reporting a history of foot ulcer in this population-based study exceeded the proportion of foot ulcer history reported previously. Height as a correlate has been occasionally reported in previous studies and needs further attention. Associated factors for a foot ulcer history help identify individuals who may be at particular risk of this adverse outcome. PMID- 18426788 TI - Stroke and acute myocardial infarction in the Swedish Sami population: incidence and mortality in relation to income and level of education. AB - BACKGROUND: Gender differences in cardiovascular diseases (CVD) among the Sami have been reported previously. The aim of the present study was to investigate the incidence of and mortality from stroke, subarachnoid haemorrhage (SAH), and acute myocardial infarction (AMI) in the Swedish Sami population between 1985 and 2002, and to analyse the potential impact of income and level of education on cardiovascular morbidity and mortality. METHODS: A Sami cohort of 15,914 persons (4,465 reindeer herding and 11,449 non-herding Sami) were followed up from 1985 to 2002 with regard to incidence and mortality rates of AMI, stroke, and SAH. Incidence and mortality ratios were calculated using a demographically matched non-Sami control population (DMC) as the standard (71,550 persons). RESULTS: There was no elevated risk of developing AMI among the Sami compared with the DMC. However, the mortality ratio of AMI was significantly higher for Sami women. Higher incidence rates of stroke and SAH for both Sami men and women was observed, but no differences in mortality rates. Apart from the reindeer-herding men who demonstrated lower levels of income and education, the income and education levels among Sami were similar to the DMC. CONCLUSIONS: High mortality rates from AMI rather than stroke explain the excess mortality for CVD previously shown among Sami women. The results suggest that the differences in incidence of stroke between herding and non-herding Sami men, and between Sami women and non Sami women, are caused by behavioural and psychosocial risk factors rather than by traditional socioeconomic ones. PMID- 18426789 TI - When the helper humiliates the patient: a qualitative study about unintended intimidations. AB - AIMS: To explore experiences of intimidating patients/clients from the perspective of the professional. METHODS: This was a qualitative study based on memory work, a procedure whereby memories are converted into written stories. The setting was a workshop about oppression and empowerment in a rehabilitation conference. Participants were 48 health and social professionals in six groups. Thirty-seven of the participants provided their written stories. The main outcome measures were descriptions of intimidation episodes according to the level of passivity/activity in the professional responsible for the act. RESULTS: The participants were able to recall episodes where they later realized that they had been involved in events perceived as intimidations by the patient/client. The memory stories described events that were not isolated events, but represented issues that might occur now and then. The first type of intimidation included events where the professional intimidated the patient/client by neglecting various basic needs, such as bodily functions, information, regard, time, and patience. The second type included events where the professional patronized and overruled the patient/client by pursuing the professional's agenda, ridiculing, or only partially acknowledging, the person. Remembering and presenting these events created ideas for alternative modes of action for the professionals. CONCLUSIONS: Systems for health and social care suffer from lack of resources, time, and proximity -- issues that facilitate the occurrence of intimidation of patients/clients. Yet, the professional remains responsible for his or her moral conduct. High moral awareness and systems with a better balance between reason and emotion are needed to stop vulnerable people being subjected to disempowering practices. PMID- 18426790 TI - News on health policy and public health. Nordic Ministers of Health: important decisions or events in 2007. Paula Risikko, Minister of Health and Social Services, Finland. Gudlaugur Thor Thordarson, Health Minister, Iceland. PMID- 18426791 TI - Protein profiling of human breast tumor cells identifies novel biomarkers associated with molecular subtypes. AB - Molecular subtypes of breast cancer with relevant biological and clinical features have been defined recently, notably ERBB2-overexpressing, basal-like, and luminal-like subtypes. To investigate the ability of mass spectrometry-based proteomics technologies to analyze the molecular complexity of human breast cancer, we performed a SELDI-TOF MS-based protein profiling of human breast cell lines (BCLs). Triton-soluble proteins from 27 BCLs were incubated with ProteinChip arrays and subjected to SELDI analysis. Unsupervised global hierarchical clustering spontaneously discriminated two groups of BCLs corresponding to "luminal-like" cell lines and to "basal-like" cell lines, respectively. These groups of BCLs were also different in terms of estrogen receptor status as well as expression of epidermal growth factor receptor and other basal markers. Supervised analysis revealed various protein biomarkers with differential expression in basal-like versus luminal-like cell lines. We identified two of them as a carboxyl terminus-truncated form of ubiquitin and S100A9. In a small series of frozen human breast tumors, we confirmed that carboxyl terminus-truncated ubiquitin is observed in primary breast samples, and our results suggest its higher expression in luminal-like tumors. S100A9 up regulation was found as part of the transcriptionally defined basal-like cluster in DNA microarrays analysis of human tumors. S100A9 association with basal subtypes as well as its poor prognosis value was demonstrated on a series of 547 tumor samples from early breast cancer deposited in a tissue microarray. Our study shows the potential of integrated genomics and proteomics profiling to improve molecular knowledge of complex tumor phenotypes and identify biomarkers with valuable diagnostic or prognostic values. PMID- 18426792 TI - AMP-activated protein kinase phosphorylates and desensitizes smooth muscle myosin light chain kinase. AB - Smooth muscle contraction is initiated by a rise in intracellular calcium, leading to activation of smooth muscle myosin light chain kinase (MLCK) via calcium/calmodulin (CaM). Activated MLCK then phosphorylates the regulatory myosin light chains, triggering cross-bridge cycling and contraction. Here, we show that MLCK is a substrate of AMP-activated protein kinase (AMPK). The phosphorylation site in chicken MLCK was identified by mass spectrometry to be located in the CaM-binding domain at Ser(815). Phosphorylation by AMPK desensitized MLCK by increasing the concentration of CaM required for half maximal activation. In primary cultures of rat aortic smooth muscle cells, vasoconstrictors activated AMPK in a calcium-dependent manner via CaM-dependent protein kinase kinase-beta, a known upstream kinase of AMPK. Indeed, vasoconstrictor-induced AMPK activation was abrogated by the STO-609 CaM dependent protein kinase kinase-beta inhibitor. Myosin light chain phosphorylation was increased under these conditions, suggesting that contraction would be potentiated by ablation of AMPK. Indeed, in aortic rings from mice in which alpha1, the major catalytic subunit isoform in arterial smooth muscle, had been deleted, KCl- or phenylephrine-induced contraction was increased. The findings suggest that AMPK attenuates contraction by phosphorylating and inactivating MLCK. This might contribute to reduced ATP turnover in the tonic phase of smooth muscle contraction. PMID- 18426793 TI - Molecular architecture of the major histocompatibility complex class I-binding site of Ly49 natural killer cell receptors. AB - Natural killer (NK) cells play a vital role in the detection and destruction of virally infected and tumor cells during innate immune responses. The highly polymorphic Ly49 family of NK receptors regulates NK cell function by sensing major histocompatibility complex class I (MHC-I) molecules on target cells. Despite the determination of two Ly49-MHC-I complex structures, the molecular features of Ly49 receptors that confer specificity for particular MHC-I alleles have not been identified. To understand the functional architecture of Ly49 binding sites, we determined the crystal structures of Ly49C and Ly49G and completed refinement of the Ly49C-H-2K(b) complex. This information, combined with mutational analysis of Ly49A, permitted a structure-based classification of Ly49s that we used to dissect the binding site into three distinct regions, each having different roles in MHC recognition. One region, located at the center of the binding site, has a similar structure across the Ly49 family and mediates conserved interactions with MHC-I that contribute most to binding. However, the preference of individual Ly49s for particular MHC-I molecules is governed by two regions that flank the central region and are structurally more variable. One of the flanking regions divides Ly49s into those that recognize both H-2D and H-2K versus only H-2D ligands, whereas the other discriminates among H-2D or H-2K alleles. The modular design of Ly49-binding sites provides a framework for predicting the MHC-binding specificity of Ly49s that have not been characterized experimentally. PMID- 18426794 TI - The RalA GTPase is a central regulator of insulin exocytosis from pancreatic islet beta cells. AB - RalA is a small GTPase that is thought to facilitate exocytosis through its direct interaction with the mammalian exocyst complex. In this study, we report an essential role for RalA in regulated insulin secretion from pancreatic beta cells. We employed lentiviral-mediated delivery of RalA short hairpin RNAs to deplete endogenous RalA protein in mouse pancreatic islets and INS-1 beta cells. Perifusion of mouse islets depleted of RalA protein exhibited inhibition of both first and second phases of glucose-stimulated insulin secretion. Consistently, INS-1 cells depleted of RalA caused a severe inhibition of depolarization-induced insulin exocytosis determined by membrane capacitance, including a reduction in the size of the ready-releasable pool of insulin granules and a reduction in the subsequent mobilization and exocytosis of the reserve pool of granules. Collectively, these data suggest that RalA is a critical component in biphasic insulin release from pancreatic beta cells. PMID- 18426795 TI - Independent generation of Abeta42 and Abeta38 peptide species by gamma-secretase. AB - Proteolytic processing of the amyloid precursor protein by beta- and gamma secretase generates the amyloid-beta (Abeta) peptides, which are principal drug targets in Alzheimer disease therapeutics. gamma-Secretase has imprecise cleavage specificity and generates the most abundant Abeta40 and Abeta42 species together with longer and shorter peptides such as Abeta38. Several mechanisms could explain the production of multiple Abeta peptides by gamma-secretase, including sequential processing of longer into shorter Abeta peptides. A novel class of gamma-secretase modulators (GSMs) that includes some non-steroidal anti inflammatory drugs has been shown to selectively lower Abeta42 levels without a change in Abeta40 levels. A signature of GSMs is the concomitant increase in shorter Abeta peptides, such as Abeta38, leading to the suggestion that generation of Abeta42 and Abeta38 peptide species by gamma-secretase is coordinately regulated. However, no evidence for or against such a precursor product relationship has been provided. We have previously shown that stable overexpression of aggressive presenilin-1 (PS1) mutations associated with early onset familial Alzheimer disease attenuated the cellular response to GSMs, resulting in greatly diminished Abeta42 reductions as compared with wild type PS1. We have now used this model system to investigate whether Abeta38 production would be similarly affected indicating coupled generation of Abeta42 and Abeta38 peptides. Surprisingly, treatment with the GSM sulindac sulfide increased Abeta38 production to similar levels in four different PS1 mutant cell lines as compared with wild type PS1 cells. This was confirmed with the structurally divergent GSMs ibuprofen and indomethacin. Mass spectrometry analysis and high resolution urea gel electrophoresis further demonstrated that sulindac sulfide did not induce detectable compensatory changes in levels of other Abeta peptide species. These data provide evidence that Abeta42 and Abeta38 species can be independently generated by gamma-secretase and argue against a precursor-product relationship between these peptides. PMID- 18426796 TI - Endoplasmic reticulum (ER) chaperone regulation and survival of cells compensating for deficiency in the ER stress response kinase, PERK. AB - The activity of PERK, an endoplasmic reticulum (ER) transmembrane protein kinase, assists in an ER stress response designed to inhibit general protein synthesis while allowing upregulated synthesis of selective proteins such as the ATF4 transcription factor. PERK null mice exhibit phenotypes that especially affect secretory cell types. Although embryonic fibroblasts from these mice are difficult to transfect with high efficiency, we have generated 293 cells stably expressing the PERK-K618A dominant negative mutant. 293/PERK-K618A cells, in response to ER stress: (a) do not properly inhibit general protein synthesis, (b) exhibit defective/delayed induction of ATF4 and BiP, and (c) exhibit exuberant splice activation of XBP1 and robust cleavage activation of ATF6, with abnormal regulation of calreticulin levels. The data suggest compensatory mechanisms allowing for cell survival in the absence of functional PERK. Interestingly, although induction of CHOP (a transcription factor implicated in apoptosis) is notably delayed after onset of ER stress, 293/PERK-K618A cells eventually produce CHOP at normal or even supranormal levels and exhibit increased apoptosis either in response to general ER stress or, more importantly, to specific misfolded secretory proteins. PMID- 18426798 TI - An intracellular interaction network regulates conformational transitions in the dopamine transporter. AB - Neurotransmitter:sodium symporters (NSS)(1) mediate sodium-dependent reuptake of neurotransmitters from the synaptic cleft and are targets for many psychoactive drugs. The crystal structure of the prokaryotic NSS protein, LeuT, was recently solved at high resolution; however, the mechanistic details of regulation of the permeation pathway in this class of proteins remain unknown. Here we combine computational modeling and experimental probing in the dopamine transporter (DAT) to demonstrate the functional importance of a conserved intracellular interaction network. Our data suggest that a salt bridge between Arg-60 in the N terminus close to the cytoplasmic end of transmembrane segment (TM) 1 and Asp-436 at the cytoplasmic end of TM8 is stabilized by a cation-pi interaction between Arg-60 and Tyr-335 at the cytoplasmic end of TM6. Computational probing illustrates how the interactions may determine the flexibility of the permeation pathway, and mutagenesis within the network and results from assays of transport, as well as the state-dependent accessibility of a substituted cysteine in TM3, support the role of this network in regulating access between the substrate binding site and the intracellular milieu. The mechanism that emerges from these findings may be unique to the NSS family, where the local disruption of ionic interactions modulates the transition of the transporter between the outward- and inward facing conformations. PMID- 18426797 TI - Functional interaction between paramyxovirus fusion and attachment proteins. AB - Paramyxovirinae envelope glycoproteins constitute a premier model to dissect how specific and dynamic interactions in multisubunit membrane protein complexes can control deep-seated conformational rearrangements. However, individual residues that determine reciprocal specificity of the viral attachment and fusion (F) proteins have not been identified. We have developed an assay based on a pair of canine distemper virus (CDV) F proteins (strains Onderstepoort (ODP) and Lederle) that share approximately 95% identity but differ in their ability to form functional complexes with the measles virus (MV) attachment protein (H). Characterization of CDV F chimeras and mutagenesis reveals four residues in CDV F ODP (positions 164, 219, 233, and 317) required for productive interaction with MV H. Mutating these residues to the Lederle type disrupts triggering of F-ODP by MV H without affecting functionality when co-expressed with CDV H. Co immunoprecipitation shows a stronger physical interaction of F-ODP than F-Lederle with MV H. Mutagenesis of MV F highlights the MV residues homologous to CDV F residues 233 and 317 as determinants for physical glycoprotein interaction and fusion activity under homotypic conditions. In assay reversal, the introduction of sections of the CDV H stalk into MV H shows a five-residue fragment (residues 110-114) to mediate specificity for CDV F-Lederle. All of the MV H stalk chimeras are surface-expressed, show hemadsorption activity, and trigger MV F. Combining the five-residue H chimera with the CDV F-ODP quadruple mutant partially restores activity, indicating that the residues identified in either glycoprotein contribute interdependently to the formation of functional complexes. Their localization in structural models of F and H suggests that placement in particular of F residue 233 in close proximity to the 110-114 region of H is structurally conceivable. PMID- 18426799 TI - Nitrosative stress leads to protein glutathiolation, increased s-nitrosation, and up-regulation of peroxiredoxins in the heart. AB - Nitric oxide (NO) is produced by different isoforms of nitric oxide synthases (NOSs) and operates as a mediator of important cell signaling pathways, such as the cGMP signaling cascade. Another mechanism by which NO exerts biological effects is mediated through S-nitrosation of target proteins. To explore thiol based protein modifications in a situation of defined nitrosative stress, we used a transgenic mouse model with cardiac specific overexpression of inducible nitric oxide synthase (iNOS) and concomitant myoglobin deficiency (iNOS(+)/myo(-/-)). In comparison with the wild type hearts, protein glutathiolation detected by immunoblotting was significantly enhanced in iNOS(+)/myo(-/-) hearts, whereas protein S-nitrosation as measured by the biotin switch assay and two-dimensional PAGE revealed that nearly all of the detected proteins ( approximately 60) remained unchanged with the exception of three proteins. Tandem mass spectrometry revealed these proteins to be peroxiredoxins (Prxs), which are known to possess peroxidase activity, whereby hydrogen peroxide, peroxynitrite, and a wide range of organic hydroperoxides are reduced and detoxified. Immunoblotting with specific antibodies revealed up-regulation of Prx VI in the iNOS(+)/myo(-/-) hearts, whereas expression of Prx II and Prx III remained unchanged. Furthermore, the analysis of the cardiac S-nitrososubproteome identified several new proteins possibly being involved in NO-signaling pathways. Our data indicate that S nitrosation and glutathiolation of cardiac proteins may contribute to the phenotype of NO-induced heart failure. The up-regulation of antioxidant proteins like Prx VI appears to be an additional mechanism to antagonize an excess of reactive oxygen/nitrogen species. Furthermore, S-nitrosation of Prxs may serve a new function in the signaling cascade of nitrosative stress. PMID- 18426801 TI - The functional interaction of 14-3-3 proteins with the ERK1/2 scaffold KSR1 occurs in an isoform-specific manner. AB - Identifying 14-3-3 isoform-specific substrates and functions may be of broad relevance to cell signaling research because of the key role played by this family of proteins in many vital processes. A multitude of ligands have been identified, but the extent to which they are isoform-specific is a matter of debate. Herein we demonstrate, both in vitro and in vivo, a specific, functionally relevant interaction of human 14-3-3gamma with the molecular scaffold KSR1, which is mediated by the C-terminal stretch of 14-3-3gamma. Specific binding to 14-3-3gamma protected KSR1 from epidermal growth factor induced dephosphorylation and impaired its ability to activate ERK2 and facilitate Ras signaling in Xenopus oocytes. Furthermore, RNA interference mediated inhibition of 14-3-3gamma resulted in the accumulation of KSR1 in the plasma membrane, all in accordance with 14-3-3gamma being the cytosolic anchor that keeps KSR1 inactive. We also provide evidence that KSR1-bound 14-3-3gamma heterodimerized preferentially with selected isoforms and that KSR1 bound monomeric 14-3-3gamma. In sum, we have demonstrated ligand discrimination among 14-3-3 isoforms and shed light on molecular mechanisms of 14-3-3 functional specificity and KSR1 regulation. PMID- 18426800 TI - The PIM1 kinase is a critical component of a survival pathway activated by docetaxel and promotes survival of docetaxel-treated prostate cancer cells. AB - A defining characteristic of solid tumors is the capacity to divide aggressively and disseminate under conditions of nutrient deprivation, limited oxygen availability, and exposure to cytotoxic drugs or radiation. Survival pathways are activated within tumor cells to cope with these ambient stresses. We here describe a survival pathway activated by the anti-cancer drug docetaxel in prostate cancer cells. Docetaxel activates STAT3 phosphorylation and transcriptional activity, which in turns induces expression of the PIM1 gene, encoding a serine-threonine kinase activated by many cellular stresses. Expression of PIM1 improves survival of docetaxel-treated prostate cancer cells, and PIM1 knockdown or expression of a dominant-negative PIM1 protein sensitize cells to the cytotoxic effects of docetaxel. PIM1 in turn mediates docetaxel induced activation of NFkappaB transcriptional activity, and PIM1 depends in part on RELA/p65 proteins for its prosurvival effects. The PIM1 kinase plays a critical role in this STAT3 --> PIM1 --> NFkappaB stress response pathway and serves as a target for intervention to enhance the therapeutic effects of cytotoxic drugs such as docetaxel. PMID- 18426802 TI - Induction of cytosolic calcium flux by CD20 is dependent upon B Cell antigen receptor signaling. AB - The anti-CD20 monoclonal antibody (mAb) rituximab is now routinely used for the treatment of non-Hodgkins lymphoma and is being examined in a wide range of other B-cell disorders, such as rheumatoid arthritis. Despite intensive study, the mechanism of action still remains uncertain. In the current study, anti-CD20 mAb induced calcium signaling was investigated. Previously, we grouped anti-CD20 mAbs into Type I (rituximab-like) and Type II (B1-like) based upon various characteristics such as their ability to induce complement activation and redistribute CD20 into detergent-insoluble membrane domains. Here we show that only Type I mAbs are capable of inducing a calcium flux in B cells and that this is tightly correlated with the expression of the B-cell antigen receptor (BCR). Inhibitor analysis revealed that the signaling cascade employed by CD20 was strikingly similar to that utilized by the BCR, with inhibitors of Syk, Src, and PI3K, but not EGTA, p38, or ERK1/2, completely ablating calcium flux. Furthermore, binding of Type I but not Type II mAbs caused direct association of CD20 with the BCR as measured by FRET and resulted in the phosphorylation of BCR specific adaptor proteins BLNK and SLP-76. Crucially, variant Ramos cells lacking BCR expression but with unchanged CD20 expression were completely unable to induce calcium flux following ligation of CD20. Collectively, these data indicate that CD20 induces cytosolic calcium flux through its ability to associate with and "hijack" the signaling potential of the BCR. PMID- 18426803 TI - A structural model of the GDP dissociation inhibitor rab membrane extraction mechanism. AB - Rab GDP dissociation inhibitors (GDI)-facilitated extraction of prenylated Rab proteins from membranes plays an important role in vesicular membrane trafficking. The investigated thermodynamic properties of yeast Rab.GDI and Rab.MRS6 complexes demonstrated differences in the Rab binding properties of the closely related Rab GDI and MRS6 proteins, consistent with their functional diversity. The importance of the Rab C terminus and its prenylation for GDI/MRS6 binding was demonstrated using both biochemical and structural data. The presented structures of the apo-form yeast Rab GDI and its two complexes with unprenylated Rab proteins, together with the earlier published structures of the prenylated Ypt1.GDI, provide evidence of allosteric regulation of the GDI lipid binding site opening, which plays a key role in the proposed mechanism of GDI mediated Rab extraction. We suggest a model for the interaction of GDI with prenylated Rab proteins that incorporates a stepwise increase in affinity as the three different partial interactions are successively formed. PMID- 18426804 TI - Comparative conservation analysis of the human mitotic phosphoproteome. AB - MOTIVATION: A key challenge in phosphoproteomic studies is to distinguish functionally relevant phosphorylation sites from potentially 'silent' phosphorylation. Considering that relevant phosphorylation sites are expected to be better conserved during evolution than overall Serine, Threonine and Tyrosine (S/ T/ Y) residues, we asked whether this can be directly demonstrated through statistic analysis, using a large experimental dataset. RESULTS: Analyzing phosphoproteomic data derived from the human mitotic spindle apparatus, we found that 95.2% of 1744 phosphorylation sites are conserved in at least one of six other vertebrate species. Using a new score, termed conservation Z-score (CZ score), we demonstrate that phosphorylation sites are significantly better conserved than other S/T/Y sites, a conclusion validated from several kinase consensus motifs. Most importantly, phosphorylation sites with experimentally verified biological functions were significantly better conserved than other phosphorylation sites, indicating that analysis utilizing evolutionary conservation may constitute a powerful basis for the development of improved phosphorylation site predictors. CONTACT: malik@biochem.mpg.de SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online. PMID- 18426805 TI - Prediction of disordered regions in proteins based on the meta approach. AB - MOTIVATION: Intrinsically disordered regions in proteins have no unique stable structures without their partner molecules, thus these regions sometimes prevent high-quality structure determination. Furthermore, proteins with disordered regions are often involved in important biological processes, and the disordered regions are considered to play important roles in molecular interactions. Therefore, identifying disordered regions is important to obtain high-resolution structural information and to understand the functional aspects of these proteins. RESULTS: We developed a new prediction method for disordered regions in proteins based on the meta approach and implemented a web-server for this prediction method named 'metaPrDOS'. The method predicts the disorder tendency of each residue using support vector machines from the prediction results of the seven independent predictors. Evaluation of the meta approach was performed using the CASP7 prediction targets to avoid an overestimation due to the inclusion of proteins used in the training set of some component predictors. As a result, the meta approach achieved higher prediction accuracy than all methods participating in CASP7. PMID- 18426806 TI - Eukaryotic transcription factor binding sites--modeling and integrative search methods. AB - A comprehensive knowledge of transcription factor binding sites (TFBS) is important for a mechanistic understanding of transcriptional regulation as well as for inferring gene regulatory networks. Because the DNA motif recognized by a transcription factor is typically short and degenerate, computational approaches for identifying binding sites based only on the sequence motif inevitably suffer from high error rates. Current state-of-the-art techniques for improving computational identification of binding sites can be broadly categorized into two classes: (1) approaches that aim to improve binding motif models by extracting maximal sequence information from experimentally determined binding sites and (2) approaches that supplement binding motif models with additional genomic or other attributes (such as evolutionary conservation). In this review we will discuss recent attempts to improve computational identification of TFBS through these two types of approaches and conclude with thoughts on future development. PMID- 18426807 TI - Separation of light-induced linear, cyclic and stroma-sourced electron fluxes to P700+ in cucumber leaf discs after pre-illumination at a chilling temperature. AB - Pre-illumination of cucumber leaf discs at 4 degrees C with low-irradiance white light (i) led to a marked decrease in the extent of photo-oxidation of P700 (the special chlorophyll pair in the PSI reaction center) in actinic light at room temperature and (ii) hastened the post-illumination re-reduction of P700+. Quantifying the linear, cyclic and stroma-sourced electron fluxes to P700+ in two actinic light regimes, we found that there was no increase in cyclic or linear electron fluxes to account for these changes. Rather, we observed a decrease in the maximum extent of P700 photo-oxidation assayed by a strong flash superimposed on continuous, background light of wavelength 723 nm, which we interpret to represent a loss of stable charge separation in PSI due to enhanced charge recombination as a result of the pre-illumination treatment. The funneling of electrons towards fewer non-damaged PSI complexes could explain the hastened post illumination re-reduction of P700+, aided by a slight increase in a stroma sourced electron flux after prolonged pre-illumination at 4 degrees C. Quantifying the separate fluxes to P700+ helps to elucidate the effects of chilling of cucumber leaf discs in the light and the reasons for the hastened post-illumination re-reduction of P700+. PMID- 18426808 TI - John Bryant takes a closer look at some of this month's Original Articles. PMID- 18426810 TI - Characterization of a novel complex BRAF mutation in a follicular variant papillary thyroid carcinoma. AB - INTRODUCTION: Activating mutations of the BRAF oncogene are frequently detected in papillary thyroid carcinoma (PTC) and have been associated with a worse prognosis. The amino acid substitution V600E accounts for 90% of all oncogenic BRAF mutations and is typically detected in classic PTCs, whereas other less frequent BRAF mutations seem to be associated with other PTC histotypes. CASE: Screening for activating BRAF mutations in a series of 83 PTCs identified the most common V600E mutation in 39 cases (histologically, 38 classic PTCs and 1 sclerosing variant PTC) and a complex in-frame mutation involving amino acids V600-S605 in a stage III multicentric follicular variant PTC, occurring in a 50 year-old female patient, who was affected by hypothyroidism in autoimmune thyroiditis and had a family history of PTC and autoimmune thyroiditis. Since the identified BRAF mutation was novel in the literature, bioinformatic modeling was performed to predict its impact on BRAF activity. Although the mutation resulted in loss of a phosphorylation site in the activation loop of BRAF, it was predicted to increase BRAF kinase activity by mimicking an activating phosphorylation. CONCLUSIONS: This study, which reports a new BRAF mutation, highlights the usefulness of bioinformatic modeling in the prediction of functional effects of new mutations and indicates that mutation-specific screening tests might miss some rare BRAF mutations. These facts should be taken into consideration in the molecular diagnosis of thyroid cancer and in the design of therapeutic protocols based on inhibitors of the BRAF pathway. PMID- 18426812 TI - Relation of 25-hydroxyvitamin D and parathyroid hormone levels with metabolic syndrome among US adults. AB - OBJECTIVE: Previous research on the combined association of 25-hydroxyvitamin D (25(OH)D) and parathyroid hormone (PTH) with metabolic syndrome may have been limited by restricted age variability and a lack of representation of the general population. This study examined the combined association of 25(OH)D and PTH with Adult Treatment Panel III-defined MetSyn among a nationally representative sample of US adults. DESIGN AND METHODS: This population-based cross-sectional study included 834 men and 820 women aged > or =20 years without diagnosed diabetes who completed a physical examination as part of the 2003-2004 US National Health and Nutrition Examination Survey. RESULTS: After adjusting for age, sex, race/ethnicity, income, lifestyle factors, total calcium, and energy intake, the odds ratio (OR) for MetSyn in the highest quintile of 25(OH)D (median 88.0 nmol/l) compared with the lowest quintile (median 26.8 nmol/l) was 0.27 (0.15, 0.46; P(trend)<0.001). This relation was unchanged after additional adjustment for PTH level (OR, 0.26; 0.15, 0.44; P(trend)<0.001) and did not differ by sex (P interaction 0.6) or age (< or > or =50 years; P interaction 0.2). In contrast, the multivariable-adjusted odds for MetSyn increased with increasing PTH among older men (P(trend) 0.004), but not younger men (P(trend) 0.4) or women regardless of age (P(trend) 0.4 in younger and older women). CONCLUSIONS: These data suggest an inverse association of 25(OH)D with MetSyn, independent of potential confounding factors, calcium intake, and PTH, and a positive association of PTH with MetSyn among older men. PMID- 18426811 TI - Heterogeneity in the molecular basis of ACTH resistance syndrome. AB - OBJECTIVE: ACTH resistance syndromes are rare, autosomal, and genetically heterogeneous diseases that include familial glucocorticoid deficiency (FGD) and triple A syndrome. FGD has been shown to segregate with mutations in the gene coding for ACTH receptor (MC2R) or melanocortin 2 receptor accessory protein (MRAP), whereas mutations in the triple A syndrome (AAAS, Allgrove syndrome) gene have been found in segregation with triple A syndrome. We describe the clinical findings and molecular analysis of MC2R, MRAP, and AAAS genes in five Brazilian patients with ACTH resistance syndrome. DESIGN AND METHODS: Genomic DNA from patients and their unaffected relatives was extracted from peripheral blood leucocytes and amplified by PCR, followed by automated sequencing. Functional analysis was carried out using Y6 cells expressing wild-type and mutant MC2R. RESULTS: All five patients showed low cortisol and elevated plasma ACTH levels. One patient had achalasia and alacrima, besides the symptoms of adrenal insufficiency. The molecular analysis of FGD patients revealed a novel p.Gly116Val mutation in the MC2R gene in one patient and p.Met1Ile mutation in the MRAP gene in another patient. Expression of p.Gly116Val MC2R mutant in Y6 cells revealed that this variant failed to stimulate cAMP production. The analysis of the AAAS gene in the patient with triple A syndrome showed a novel g.782_783delTG deletion. The molecular analysis of DNA from other two patients showed no mutation in MC2R, MRAP, or AAAS gene. CONCLUSIONS: In conclusion, the molecular basis of ACTH resistance syndrome is heterogeneous, segregating with genes coding for proteins involved with ACTH receptor signaling/expression or adrenal gland development and other unknown genes. PMID- 18426813 TI - The methylenetetrahydrofolate reductase C677T genotype and the risk of obesity in three large population-based cohorts. AB - OBJECTIVE: Epidemiological studies have shown that low folate levels are associated with a high body mass index (BMI). These findings have potentially important health implications and warrant further investigation to determine whether a causal relationship exists and the direction of this relationship. The methylenetetrahydrofolate reductase (MTHFR) C677T TT genotype is associated with reduced folate availability and may be a surrogate for measuring folate levels. We sought to determine whether MTHFR C677T genotype was associated with obesity. DESIGN: We carried out our study on four populations from three longitudinal studies based in the UK and Denmark in which DNA for genotyping was obtained along with measures of obesity. METHODS: Our subjects were taken from the British Women's Heart and Health Study (BWHHS), the Avon Longitudinal Study of Parents and Children (two populations: mothers and children) and the Copenhagen City Heart Study. We performed analyses separately by population, and then carried out a meta-analysis, combining similar populations. RESULTS: Initial findings in the BWHHS suggested that the TT genotype may be associated with an increased risk of obesity BMI > or =30, however, no association was found with BMI or central adiposity in this cohort. This genotype was not associated with obesity in our other cohorts. CONCLUSIONS: Our results suggest that the initial positive finding with obesity in the BWHHS was a chance finding. Our findings do not support a causal effect of low folate on obesity. PMID- 18426814 TI - Retinol-binding protein-4 is not strongly associated with insulin sensitivity in normal pregnancies. AB - OBJECTIVE: Recently, experimental and clinical studies suggest that retinol binding protein-4 (RBP4) may provide a link between obesity and insulin resistance. However, no previous studies have investigated the impact of circulating RBP4 on measures of insulin resistance in normal pregnant women, and the objective of this study is to measure serum RBP4 in early and late pregnancy and relate these to measures of insulin resistance and secretion controlling for changes in fat mass. DESIGN AND METHODS: Samples were obtained during oral glucose tolerance test (OGTT) from 44 normal pregnancies at weeks 14-16 and 30 32. Measures of fat mass were body mass index (BMI) and leptin while insulin sensitivity and secretion were predicted from OGTT. Leptin and RPB4 were measured by immunoassay. RESULTS: Insulin sensitivity decreased during the course of pregnancy. Insulin sensitivity and secretion were best explained by BMI and circulating leptin, but not RBP4, both in early and late pregnancy. However, a marked increase in fasting RBP4 from early to late pregnancy was observed, and this change was associated with a decline in insulin sensitivity. A marked increase in RBP4 was found during OGTT at weeks 14-16 with an opposite temporal course at weeks 30-32. CONCLUSION: The increased fat mass and insulin resistance during normal pregnancy was best explained by measures of fat mass. However, the increase in RBP4 from early to late pregnancy, associated with a decline in insulin sensitivity, potentially indicates interactions with glucose metabolism. PMID- 18426815 TI - RANTES/CCL5 gene polymorphisms, serum concentrations, and incident type 2 diabetes: results from the MONICA/KORA Augsburg case-cohort study, 1984-2002. AB - OBJECTIVE: Regulated on activation, normal T-cell expressed and secreted (RANTES)/chemokine(C-C motif) ligand (CCL5) is expressed by adipocytes, and serum levels of RANTES are increased in obesity and type 2 diabetes. The aim of this study was to test the hypothesis that RANTES is involved in the pathogenesis of type 2 diabetes by analyzing the triangular association between CCL5 gene polymorphisms, systemic RANTES concentrations, and incident type 2 diabetes in a large prospective study. SUBJECTS AND METHODS: The study is based on 502 individuals (293 men and 209 women) and 1632 individuals (859 men and 773 women) with and without incident type 2 diabetes from the population-based MONItoring of Trends and Determinants in Cardiovascular Disease (MONICA)/Cooperative Health Research in the Region of Augsburg (KORA) case-cohort study respectively (mean follow-up time+/-s.d. 10.1+/-4.9 years). CCL5 genotypes and RANTES serum concentrations were determined and associations between genotypes, haplotypes, serum levels, and incident type 2 diabetes were assessed. Results Minor alleles of four single nucleotide polymorphisms were associated with lower RANTES levels (P(additive) between 1.2 x 10(-9) and 3.1 x 10(-8)), but neither genotypes, haplotypes, nor serum levels were associated with incident type 2 diabetes. CONCLUSIONS: Our data suggest that RANTES/CCL5 gene variants and serum levels are not causally related with type 2 diabetes and that elevated RANTES levels in patients with type 2 diabetes may be a consequence of hyperglycemia. However, our findings cannot preclude a local role in adipose tissue where RANTES expression may contribute to leukocyte infiltration and a proinflammatory state. PMID- 18426816 TI - Impact of musculoskeletal disease on quality of life in long-standing acromegaly. AB - OBJECTIVE: To provide rheumatological assessment of patients with long-standing acromegaly and investigate the impact of musculoskeletal disease on quality of life. DESIGN: Cross-sectional observational study. METHODS: Fifty-eight patients diagnosed with acromegaly at least 5 years previously were interviewed and examined by a rheumatologist. Each patient completed the short form-36 (SF-36), arthritis impact measurement scales 2 (AIMS2) and acromegaly quality of life questionnaires (AcroQol). RESULTS: Fifty-two out of 58 (90%) reported musculoskeletal pain, with 29 (50%) reporting neck pain. Hip osteoarthritis was present in 49 (84%) and knee osteoarthritis in 20 (34%). Half the patients (52%) reported sleep disturbance, but only 2 (3.5%) had fibromyalgia. Ten (17.2%) had previously undergone carpal tunnel decompression. Fifty-one (88%) patients had consulted their general practioner and 31 (54%) complementary therapists. SF-36, AIMS2 and AcroQol scores were lower in patients with musculoskeletal pain. CONCLUSIONS: This study of musculoskeletal problems in patients with acromegaly reports systematic rheumatological examination, use of medical services and quality of life scores. Musculoskeletal problems should be routinely addressed in acromegaly by both endocrinologist and rheumatologist and a multidisciplinary approach taken to management. PMID- 18426817 TI - Somatostatinergic ligands in dopamine-sensitive and -resistant prolactinomas. AB - OBJECTIVE: Ten percent of patients with prolactinoma fail to respond with normalization of prolactin (PRL) and tumor shrinkage under dopamine agonist (DA) therapy. The resistance to treatment is linked to a loss of dopamine receptor 2 (D2DR). Prolactinomas express somatostatin (SST) receptor subtypes, SSTR1, 2, and 5. The aim of this study was to determine whether different SST compounds could overcome the resistance to DA in prolactinomas. DESIGN AND METHODS: The efficacy of SSTR1, SSTR2, and SSTR5 ligands; the universal SST ligand, SOM230; and the chimeric SST-DA compound, BIM-23A760, was compared with cabergoline in suppressing PRL secretion from primary cultures of ten prolactinomas (six DA responders and four DA resistant). Receptor mRNAs were assessed by quantitative PCR. RESULTS: The mean mRNA levels for D2DR, SSTR1, SSTR2, and SSTR5 were 92.3+/ 47.3, 2.2+/-1.4, 1.1+/-0.7, and 1.6+/-0.6 copy/copy beta-glucuronidase (beta-Gus) respectively. The SSTR1 agonist, BIM-23926, did not suppress PRL in prolactinomas. In a DA-resistant prolactinoma, it did not inhibit [(3)H]thymidine incorporation. The SSTR5 compound, BIM-23206, produced a dose-dependent inhibition of PRL release similar to that of cabergoline in three DA-sensitive prolactinomas. BIM-23A760 produced a maximal PRL inhibition superimposable to that obtained with cabergoline with a lower EC(50) (0.5+/-0.1 vs 2.5+/-1.5 pmol/l). In DA-resistant prolactinomas, BIM-23206 and SOM230 were ineffective. Cabergoline and BIM-23A760 produced a partial inhibition of PRL secretion (19+/-6 and 21+/-3% respectively). CONCLUSION: Although the SSTRs are expressed in prolactinomas, the somatostatinergic ligands analyzed do not appear to be highly effective in suppressing PRL. D2DR remains the primary target for effective treatment of prolactinomas. PMID- 18426818 TI - Association between tumoral GH-releasing peptide receptor type 1a mRNA expression and in vivo response to GH-releasing peptide-6 in ACTH-dependent Cushing's syndrome patients. AB - OBJECTIVE: GH secretagogues (GHS) produce exaggerated ACTH and cortisol responses in Cushing's disease (CD) patients, attributable to their direct action on GH releasing peptide receptor type 1a (GHSR-1a). However, there are no studies correlating the in vivo response to GHS and GHSR-1a mRNA expression in ACTH dependent Cushing's syndrome (CS) patients. The aim of this study is to correlate the patterns of ACTH and cortisol response to GH-releasing peptide-6 (GHRP-6) to GHSR-1a expression in ACTH-dependent CS patients. DESIGN: Prospective study in a tertiary referral hospital center. Fifteen CD patients and two ectopic ACTH syndrome (EAS) patients were studied. METHODS: Tumor fragments were submitted to RNA extraction, and GHSR-1a expression was studied through real-time qPCR and compared with normal tissue samples. The patients were also submitted to desmopressin test and vasopressin receptor type 1B (AVPR1B) mRNA analysis by qPCR. RESULTS: GHSR-1a expression was similar in normal pituitary samples and in corticotrophic tumor samples. GHSR-1a expression was higher in patients (CD and EAS) presenting in vivo response to GHRP-6. Higher expression of AVPR1B was observed in the EAS patients responsive to desmopressin, as well as in corticotrophic tumors, as compared with normal pituitary samples, but no correlation between AVPR1B expression and response to desmopressin was observed in the CD patients. CONCLUSIONS: Our results revealed a higher expression of GHSR 1a in the ACTH-dependent CS patients responsive to GHRP-6, suggesting an association between receptor gene expression and in vivo response to the secretagogue in both the CD and the EAS patients. PMID- 18426819 TI - Endothelial nitric oxide synthase intron 4a/b polymorphism and early atherosclerotic changes in hypopituitary GH-deficient adult patients. AB - OBJECTIVE: Endothelial nitric oxide synthase (eNOS) intron 4a/b polymorphism is associated with plasma NO concentrations and coronary artery disease/hypertension in various populations. GH deficiency in adulthood predisposes to reduced NO concentrations and premature atherosclerosis. Our aim was to determine whether intron 4a/b polymorphism of eNOS gene influences endothelial function and early atherosclerotic changes in GH-deficient hypopituitary patients. DESIGN: Thirty three hypopituitary GH-deficient patients on conventional replacement therapy other than GH and 43 age-, sex-, and body mass index (BMI)-matched controls were studied in this cross-sectional case-control study. METHODS: Early atherosclerotic changes were determined by flow-mediated dilation (FMD) of brachial artery and carotid artery intima-media thickness (IMT). eNOS4a/b polymorphism was detected by PCR. RESULTS: Hypopituitary patients had significantly higher total/low-density lipoprotein cholesterol and fat mass and lower IGF-I concentrations compared with controls. IMT was significantly higher in patients (0.777+/-0.23 vs 0.639+/-0.17 mm, P<0.01). No significant difference was observed with respect to FMD measurements. eNOS4a/b genotype frequencies were similar between patients and controls. Patients carrying 'a' allele (a/a and a/b) had significantly higher IMT compared with controls carrying 'a' allele and bb genotype (P<0.05). However, logistic regression analysis revealed that presence of hypopituitarism, age> or =45 years, and BMI> or =27.9 kg/m(2) were significant independent predictors of IMT> or =0.65 mm. CONCLUSION: No compelling data are evident to suggest that eNOS4a/b polymorphism modifies the atherosclerotic process in GH-deficient situations. A large case-control study is needed to confirm our findings. PMID- 18426820 TI - Evaluation of health-related quality of life in patients with Cushing's syndrome with a new questionnaire. AB - Chronic exposure to hypercortisolism has significant impact on patient's health and health-related quality of life (HRQoL), as demonstrated with generic questionnaires. We have developed a disease-generated questionnaire to evaluate HRQoL in patients with Cushing's syndrome (CS; CushingQoL). OBJECTIVE: Validate the CushingQoL questionnaire in patients with CS in clinical practice conditions. DESIGN: Observational, international, cross-sectional study. METHODS: A total of 125 patients were recruited by 14 investigators from Spain, France, Germany, The Netherlands, and Italy over a 2-month period. Clinical and hormonal data were collected and correlated with results of the generic short form 36 (SF-36) questionnaire, a question on self-perceived general health status and the CushingQoL score. RESULTS: A total of 107 patients were pituitary-dependent and 18 adrenal-dependent CS; 104 (83%) were females, mean age 45 years (range 20-73 years); 39 (31%) were currently hypercortisolemic; and 47 (38%) adrenal insufficient. In clinical practice, CushingQoL was feasible (117; 94% of patients fully responded to the questionnaire in a mean time of 4 min), reliable (Crohnbach's alpha=0.87), and valid (factorial analysis demonstrated unidimensionality and Rasch analysis lead to a final version with 12 items). A significant (P<0.001) correlation was observed between CushingQoL score and patients self-perceived general health status and dimensions of SF-36 (Pearson's correlation coefficient > or =0.597). Patients with current hypercortisolism scored worse (lower) than those without (44+/-22 vs 56+/-21, P=0.004). Linear regression analysis identified female gender and hypercortisolism as significant predictors for worse QoL. CONCLUSION: CushingQoL is useful to evaluate HRQoL in patients with CS and correlates with clinical parameters. PMID- 18426821 TI - Impact of metformin versus repaglinide on non-glycaemic cardiovascular risk markers related to inflammation and endothelial dysfunction in non-obese patients with type 2 diabetes. AB - OBJECTIVE: In patients with type 2 diabetes mellitus (T2DM), biomarkers reflecting inflammation and endothelial dysfunction have been linked to cardiovascular disease (CVD biomarkers) and metabolic regulation. In T2DM patients, metformin and insulin secretagogues have demonstrated equal anti hyperglycaemic potency. Here, we report the effect of metformin versus an insulin secretagogue, repaglinide, on CVD biomarkers in non-obese T2DM patients. DESIGN AND METHODS: Single-centre, double-masked, double-dummy, crossover study during 2x4 months involving 96 non-obese (body mass index< or =27 kg/m(2)) insulin-naive T2DM patients. At enrolment, previous oral hypoglycaemic agents were stopped and the patients entered a 1-month run-in on diet-only treatment. Hereafter, patients were randomized to either 2 mg repaglinide thrice daily followed by 1 g metformin twice daily or vice versa each during 4 months with a 1-month washout between interventions. RESULTS: Levels of tumour necrosis factor-alpha, plasminogen activator inhibitor-1 antigen, tissue-type plasminogen activator antigen, von Willebrand factor, soluble intercellular adhesion molecule-1 and soluble E selectin were significantly lower during metformin versus repaglinide treatments. In contrast, Amadori albumin and heart rate were higher during metformin versus repaglinide. Levels of interleukin-6, fibrinogen, soluble vascular cell adhesion molecule-1, asymmetric dimethylarginine and advanced glycation end products as well as glycaemic levels (previously reported) and 24-h blood pressure were similar between treatments. Adjustment for known macrovascular disease did not affect the between-treatment effects. CONCLUSIONS: In non-obese T2DM patients, metformin was more effective in reducing selected biomarkers reflecting inflammation and endothelial dysfunction compared with repaglinide despite similar glycaemic levels between treatments. PMID- 18426822 TI - Early or advanced stage type 2 diabetes is not accompanied by in vivo skeletal muscle mitochondrial dysfunction. AB - OBJECTIVE: Several lines of evidence support a potential role of skeletal muscle mitochondrial dysfunction in the pathogenesis of insulin resistance and/or type 2 diabetes. However, it remains to be established whether mitochondrial dysfunction represents either cause or consequence of the disease. We examined in vivo skeletal muscle mitochondrial function in early and advanced stages of type 2 diabetes, with the aim to gain insight in the proposed role of mitochondrial dysfunction in the aetiology of insulin resistance and/or type 2 diabetes. METHODS: Ten long-standing, insulin-treated type 2 diabetes patients, 11 subjects with impaired fasting glucose, impaired glucose tolerance and/or recently diagnosed type 2 diabetes, and 12 healthy, normoglycaemic controls, matched for age and body composition and with low habitual physical activity levels were studied. In vivo mitochondrial function of the vastus lateralis muscle was evaluated from post-exercise phosphocreatine (PCr) recovery kinetics using (31)P magnetic resonance spectroscopy (MRS). Intramyocellular lipid (IMCL) content was assessed in the same muscle using single-voxel (1)H MRS. RESULTS: IMCL content tended to be higher in the type 2 diabetes patients when compared with normoglycaemic controls (P=0.06). The(31)P MRS parameters for mitochondrial function, i.e. PCr and ADP recovery time constants and maximum aerobic capacity, did not differ between groups. CONCLUSIONS: The finding that in vivo skeletal muscle oxidative capacity does not differ between long-standing, insulin-treated type 2 diabetes patients, subjects with early stage type 2 diabetes and sedentary, normoglycaemic controls suggests that mitochondrial dysfunction does not necessarily represent either cause or consequence of insulin resistance and/or type 2 diabetes. PMID- 18426823 TI - Increased Nepsilon-(carboxymethyl)-lysine levels in cerebral blood vessels of diabetic patients and in a (streptozotocin-treated) rat model of diabetes mellitus. AB - OBJECTIVE: Non-enzymatic glycation of proteins and their end products (advanced glycation end products, AGE) have been implicated in the pathogenesis of diabetic complications. Our aim was to evaluate the association between diabetes mellitus (DM) and the accumulation of one of the most abundant AGEs, Nepsilon (carboxymethyl)-lysine (CML), in cerebral vessels. RESEARCH DESIGN AND METHODS: Brain tissue samples were obtained by autopsy from 20 DM patients and 13 age matched controls. In addition, we investigated brain tissue samples of seven rats after induction of diabetes with streptozotocin (STZ) and six non-diabetic control rats. We used an immunohistochemical staining method to examine the CML immunoreactivity in the cerebral vessels. RESULTS: Staining intensity of CML was significantly higher in cerebral vessels of diabetic patients than in non diabetic subjects (median of the immunohistochemical intensity score/cm(2) in the diabetic group of 0.85 (interquartile range (IQR) 0.66-1.52) vs 0.63 in the control group (IQR 0.44-0.70); P=0.002). Furthermore, there was a similar significant difference in CML staining intensity of cerebral vessels between STZ diabetic rats and non-diabetic control rats (median of the immunohistochemical intensity score/cm(2) in the diabetic group of 1.08 (IQR 0.73-1.43) vs 0.23 in the control group (IQR 0.12-0.43); P=0.003). CONCLUSIONS: Accumulation of CML modified proteins is significantly greater in the cerebral vessels of the diabetic patients than their age-matched controls. This association has been confirmed in the insulin-deficient diabetic rat model. It may be possible that the excessive accumulation of AGE-modified proteins in the cerebral vasculature alters the local environment and microcirculation and thereby contributes to the development of cognitive impairments in diabetes. Therefore, additional study on the causal link between AGE accumulation and cognitive dysfunction and the potential benefits of AGE-blocking and/or breaking compounds is indicated. PMID- 18426824 TI - Variation in the bone morphogenetic protein-2 gene: effects on fat and lean body mass in young and elderly women. AB - OBJECTIVE: Bone morphogenetic protein-2 (BMP2) plays a critical role in osteoblastogenesis and adipogenesis from osteoprogenitor cells. The balance between osteogenic and adipogenic effects is influenced by BMP2 concentration, transcription factors and age. BMP2 single nucleotide polymorphisms (SNPs) may contribute to osteoporosis risk, but the relationship between adiposity and body composition has not been explored. We investigated the relationship between BMP2 polymorphisms and body composition in young and elderly women. DESIGN: Population based association study. METHODS: Four BMP2 SNPs studied. Total body fat and lean mass measured by DEXA in two cohorts: 'PEAK-25' women aged 25 (+/-0.00) (n=993) and osteoporosis prospective risk assessment (OPRA) women aged 75 (+/-0.00) years (n=1001). RESULTS: We found no association between BMP2 SNPs and fat or lean mass, however, we observed consistent although non-significant trends. Polymorphisms, rs235767 and Ser37Ala, exerted opposing effects on most parameters of soft tissue and bone mass in both cohorts. This relationship appeared to be age specific with large differences between alleles observed (fat mass; Ser37Ala: 14.3% (PEAK-25), -3.5% (OPRA)). These initial results appear to suggest that alleles exerting a beneficial effect in young women may subsequently contribute to phenotypes associated with osteoporosis risk in elderly women. CONCLUSIONS: While further analyses in other comparative populations are necessary, in this study of almost 2000 women we observed interesting, although non-significant trends, regarding the effects of variation in the BMP2 gene on parameters of body mass. Although the exact nature of the relationship remains uncertain, we suggest that the mechanisms are influenced by age and environmental factors. PMID- 18426825 TI - Variations in the uncoupling protein-3 gene are associated with specific obesity phenotypes. AB - OBJECTIVE: Uncoupling protein 3 (UCP-3) uncouples oxidative metabolism from ATP synthesis, resulting in the production of heat instead of energy storage. Single nucleotide polymorphisms (SNPs) in UCP-3 might result in a reduced function or expression of UCP-3 and therefore lead to an increased capacity to store energy as fat. DESIGN: We conducted a population-based, cross-sectional single-center study among 400 Dutch men between 40 and 80 years. METHODS: Seven SNPs in the UCP 3 gene were genotyped by means of an allele-specific real-time TaqMan PCR. Linear regression analyses were performed to examine the independent effects of these SNPs on obesity phenotypes. RESULTS: We found a significant association between homozygosity for the minor allele of rs647126, rs1685356, and rs2075577 and an increase in body mass index (BMI; P=0.033, P=0.016, and P=0.019 respectively). Heterozygosity for rs1685354 was associated with a significant decrease in visceral fat mass (P=0.030). CONCLUSIONS: Our results suggest that genetic variations in the UCP-3 gene are associated with an increase in BMI. A plausible mechanism by which these SNPs lead to an increase in BMI is that due to these SNPs, the UCP-3 activity might be decreased. As a result, uncoupling activity may also decrease, which will lead to an increase in body weight and BMI. PMID- 18426826 TI - Sialochemical and oxidative analyses in radioactive I131-treated patients with thyroid carcinoma. AB - BACKGROUND: I131 in relatively high doses has been shown in the past to cause damaging salivary effects and oral discomfort in patients. Although lower dosage is now widely accepted, I131 may still be the source of salivary damage over the long-term and subsequent harmful effects on both the oral cavity and the gastrointestinal tract, into which the saliva is swallowed. This study examined the effects of radioactive I131 on salivary gland activity, saliva composition and oxidative profile, and related oral discomfort complaints following thyroidectomy due to carcinoma of thyroid gland. METHODS: Out of 40 consenting female post-thyroidectomy patients, 23 (mean age 50+/-4 years old) were treated with I131 while 17 (mean age 46+/-4) were not. Whole saliva from all subjects was analyzed for antioxidant and biochemical composition and flow rate. RESULTS: The salivary flow rates of both groups were similar but their composition differed considerably. Salivary superoxide dismutase enzyme (SOD), total protein, and albumin concentrations were significantly reduced in the treated patients by 40, 25, and 18% respectively (P<0.05), as were all other salivary antioxidants. Oral discomfort complaints were far more prevalent in the I131-treated patients. CONCLUSIONS: I131-dependent damage to the salivary glands was evidenced by a broad spectrum of compositional alterations and oral complaints. Reduction in salivary antioxidant status, SOD enzyme, and the uric acid molecule leaves the oral cavity less protected against oxidative stress. This is the first report of radioactive I131 treatment being harmful to salivary glands due to compromised salivary compositional and oxidative profile and oral discomfort complaints. PMID- 18426827 TI - The role of F-18-fluorodeoxyglucose positron emission tomography in the postoperative evaluation of differentiated thyroid cancer. AB - OBJECTIVES: The objective of the study was to compare F-18-fluorodeoxyglucose position emission tomography (FDG-PET) with diagnostic whole body scanning (DxWBS) and post-ablation radioiodine whole body scanning (TxWBS) and to assess its prognostic value in newly diagnosed differentiated thyroid cancer (DTC) patients, hypothesizing that FDG-PET is more likely to disclose locoregional and distant metastases. PATIENTS AND METHODS: DxWBS and FDG-PET scanning were performed in 26 newly diagnosed DTC patients who underwent thyroidectomy and TxWBS in 24 cases who had radioactive iodine ablation. The results of the FDG-PET scans were correlated with the stage of the disease and the long-term outcome of DTC. RESULTS: Overall, 18 FDG-PET scans (69.2%) were positive showing a total of 40 foci while 8 scans (30.8%) were negative. The corresponding 26 DxWBS were all positive and showed a total of 47 foci. DxWBS and TxWBS showed similar foci in the 24 patients who had ablation therapy. In contrast to the FDG-PET scans that showed uptake of 26 foci (65%) outside the thyroid bed, 45 foci (95.7%) on DxWBS were in the thyroid bed while 2 foci (4.3%) were in cervical lymph nodes and no focus was seen outside the neck area (P=0.000). There was a clear correlation between the FDG-PET results, the stage of the disease and long-term outcome; seven of the eight negative FDG-PET scans were in stage 1, while all patients with disease higher than stage 1 (six patients) had positive scans. Over a median of 30 months (10-48), seven out of eight patients (87.5%) with negative FDG-PET scans were in remission compared with only eight patients (44.4%) with positive FDG-PET (P=0.04). CONCLUSIONS: In the postoperative evaluation of DTC, compared with DxWBS and TxWBS, FDG-PET scans are more likely to reveal uptake outside the thyroid bed and to correlate with the stage of the disease and long-term outcome. PMID- 18426828 TI - Significance of low levels of thyroglobulin in fine needle aspirates from cervical lymph nodes of patients with a history of differentiated thyroid cancer. AB - OBJECTIVE: Measurement of thyroglobulin in the washout of lymph node (LN) fine needle aspirates is recommended in the follow-up of patients with differentiated thyroid cancer (DTC). The significance of low fine needle aspirates thyroglobin (FNATg) levels remains a question, which we addressed. METHOD: Prospective study comparing FNATg with FNA cytology. Exploration of 34 DTC patients (53 cervical LNs), 26 non-thyroidectomized patients with a thyroid-unrelated cervical mass (negative controls) and 13 with 21 thyroid nodules (positive controls). The 12 DTC patients (19 LNs) with a malignant FNA cytology and/or high FNATg level received LN surgery (11 patients) or I(131)-iodine treatment (1 patient) and the outcome measure was pathological or scintigraphic evidence of DTC LN metastasis. RESULTS: All 26 negative controls showed FNATg <1 ng/FNA and all 21 positive controls showed high levels of FNATg (127-210,000 ng/FNA, median 38,000). Among DTC patients in 25 LNs with a benign FNA cytology, FNATg was undetectable in 24 and low in 1 (6 ng/FNA); in 19 LNs with a malignant FNA cytology, FNATg was high in 17 (80-140,000 ng/FNA, median 7174 ng/FNA) and low in 2 (6.6 and 7.1 ng/FNA), which proved to be low Tg immunostaining oncocytic DTC metastasis; in 9 LNs with a non-informative cytology, FNATg was undetectable in 8 but 11,825 ng/FNA in 1, which proved a DTC metastasis. Measurement of FNA albumin demonstrated that contamination of FNA by serum proteins was negligible. CONCLUSION: Low FNATg levels can indicate a DTC metastasis. It cannot be related to clinically relevant levels of serum Tg. PMID- 18426829 TI - Natural course of small adrenal lesions in multiple endocrine neoplasia type 1: an endoscopic ultrasound imaging study. AB - OBJECTIVE: Adrenal lesion is one of the features of multiple endocrine neoplasia type 1 (MEN1). This study aimed to assess prevalence, natural course and clinical relevance of small adrenal lesions without clinical symptoms, endocrine activity, or mechanical problems and thus without clear indication for surgical therapy by endoscopic ultrasound (EUS). DESIGN AND METHODS: Forty-nine patients with familial MEN1 were studied. Twenty-seven of these with adrenal lesions were detected by EUS and at least two performed EUS examinations were included into a subgroup where changes in adrenal morphology were studied by measuring changes in the largest diameter of the dominant adrenal tumour. RESULTS: EUS detected adrenal lesions in 36 (73%) patients: 6 (12%) plump adrenals, 17 (35%) nodular hyperplasia, 12 (24%) adenomas and 1 (2%) cyst. Bilateral adrenal lesions were detected in 17 patients and unilateral in 19 patients. A change in the largest tumour diameter was found to be for nodular hyperplasia -0.02+/-1.41% per month (range -2.56 to 4.58%) and for adenomas -0.61+/-1.95% per month (range -6.25 to 1.15%). One patient had an adrenal cyst with significant growth. There was no evidence of carcinoma or metastatic disease during the study. CONCLUSIONS: The prevalence of adrenal lesions in MEN1 is higher than that reported earlier. Except one cystic lesion, no significant change in the tumour size was observed over a mean observation period of more than 2 years. In a typical situation, small adrenal lesions in MEN1 seem to be constant in their morphology. PMID- 18426830 TI - Autoimmune polyendocrine syndrome type I in Slovakia: relevance of screening patients with autoimmune Addison's disease. AB - BACKGROUND: Autoimmune polyendocrine syndrome type I (APS I) is a monogenic disease affecting endocrine glands and other organs due to mutations of the autoimmune regulator (AIRE) gene. There is a wide variability in clinical phenotypes in patients with APS I, which makes the diagnosis a challenge. OBJECTIVE: To screen for APS I among Slovakian patients with sporadic Addison's disease and clinical features that raised the suspicion of APS I. METHODS: All 14 exons and exon-intron boundaries of the AIRE gene were sequenced. In addition, autoantibodies specific for Addison's disease and polyendocrine syndromes were assayed. RESULTS: Using clinical criteria we identified four patients with APS I in three families. Two patients had a novel missense mutation in exon 2 (c.274C>T, p.R92W) and either the Finnish major mutation (c.769C>T) or the common 13 bp deletion (c.967-979del13bp). APS I was diagnosed in a brother of the latter after his death due to an adrenal crisis. A fourth patient had primary adrenal failure and hypoparathyroidism without AIRE mutations or APS-I specific autoantibodies. CONCLUSIONS: Four patients with APS I were found in a Slovakian cohort of Addison patients, although the lack of detectable AIRE mutations and APS I-specific autoantibodies raises uncertainty regarding the pathogenesis in one of the patients. This study demonstrates the merits of screening patients with phenotypic features or autoantibody findings that could indicate APS I, even in adult patients. It is necessary to identify APS I patients in order to provide appropriate treatment and follow-up of the various components of APS I. PMID- 18426831 TI - Body mass index and ovarian function are associated with endocrine and metabolic abnormalities in women with hyperandrogenic syndrome. AB - BACKGROUND: The aim of this study was to evaluate associations of clinical features, such as hirsutism, polycystic ovaries (PCOs), ovulatory dysfunction, and body mass index (BMI) > or =25 kg/m(2), with metabolic abnormalities in hyperandrogenic women. METHODS: Hirsutism was based on the modified Ferriman Gallwey score. Ovulatory function was classified as eumenorrhea, oligomenorrhea and amenorrhea, and PCOs were assessed using the ultrasound criteria recommended in the Rotterdam definition. An oral glucose tolerance test was performed. Different insulin resistance (IR) indices were calculated. RESULTS: Hirsute women had significantly higher BMI, DHEA sulfate (DHEAS) and free androgen index (FAI), and significantly lower values for sex hormone-binding globulin (SHBG). Women with amenorrhea were younger in comparison to women with eumenorrhea and had significantly higher values for fasting insulin (FI) and 1- and 2-h insulin levels; lower values for glucose to insulin ratio (GIR), quantitative insulin sensitivity check index (QUICKI), and SHBG. Women with PCO had significantly higher levels of LH and low-density lipoprotein (LDL), whereas high-density lipoprotein (HDL) levels were significantly lower. Women with a BMI > or =25 kg/m(2) had significantly higher values for age, fasting plasma glucose, FI, and 1- and 2-h glucose and insulin levels, homeostatic model for assessment of IR (HOMA-IR), homeostatic model for assessment of B-cell function (HOMA-B), and FAI, whereas their GIR, insulin sensitivity index, QUICKI, SHBG, and HDL were significantly lower. CONCLUSIONS: In women with hyperandrogenic syndrome, BMI> or =25 kg/m(2) and amenorrhea appear to be associated with severe endocrine and metabolic abnormalities. PMID- 18426832 TI - Association between gynecomastia and aromatase (CYP19) polymorphisms. AB - OBJECTIVE: Aromatase cytochrome P45019 (CYP19) is a key enzyme in estrogen biosynthesis, and polymorphisms within its gene are associated with an increased risk of estrogen-dependent diseases. Enhanced estrogen stimulation of breast tissue in men may lead to gynecomastia. We assessed whether intron 4 (TTTA)n repeat and TCT deletion/insertion polymorphisms and an exon 10 (3'-UTR) C/T single nucleotide polymorphism of CYP19 are associated with gynecomastia. DESIGN/METHODS: We performed a genetic association study of 100 patients referred to the endocrinological outpatient clinic with breast glandular tissue enlargement confirmed by clinical and ultrasound examinations and 99 healthy volunteers without gynecomastia. Microsatellite (TTTA)n and insertion/deletion polymorphisms were studied using capillary electrophoresis, and the C/T polymorphism in the 3'-UTR was analyzed using the TaqMan assay. RESULTS: Significantly increased risk of gynecomastia was found in subjects carrying a CYP19 exon 10 T allele that was previously related to the high aromatase activity. Frequency of the TT genotype was significantly higher in patients when compared with controls (40.6 vs 26.3%; TT versus CT and CC genotypes; P(c)<0.05). We found strong linkage disequilibrium between the alleles of studied polymorphic loci. T allele in the 3'-UTR was in linkage disequilibrium with the long alleles of the intron 4 polymorphism, mainly (TTTA)11. However, our findings did not show significant correlation of alleles having more than nine TTTA repeats with gynecomastia. CONCLUSIONS: The CYP19 polymorphisms might contribute to the incidence of gynecomastia, but further studies in larger groups are needed to confirm these results. PMID- 18426833 TI - Activating transcription factor 3: a hormone responsive gene in the etiology of hypospadias. AB - INTRODUCTION: Hypospadias is a common inborn error of the genital development, whose complex etiology remains elusive. Defects of the androgen metabolism and activity have been found in a subset of boys with hypospadias. Moreover, the balance between androgens and estrogens seems to be important to the proper male genital development. Activating transcription factor 3 (ATF3), an estrogen responsive gene, has been reported to be expressed during sexual development and up-regulated in hypospadic genital skin. We investigated ATF3 as a candidate gene for hypospadias. MATERIAL AND METHODS: Genotyping of eight-tagged single nucleotide polymorphisms (SNP)s was performed in 330 boys with hypospadias and in 380 healthy controls. Screening for mutations in ATF3 was conducted in a subset of boys with hypospadias. ATF3 expression was evaluated in the foreskin of boys with hypospadias and in healthy controls and in the human fetal genitalia by immunohistochemistry. RESULTS: Three common SNPs, spanning a region of about 16 kb in intron 1 of ATF3, are associated with hypospadias. These SNPs are not linked and their effects are independent. The combination of the three risk SNPs yields the highest significance. Mutation screening identified the gene variant c536A>G in one patient and c817C>T in the 3'-UTR in two other patients. ATF3 expression was evidenced in the developing male urethra. CONCLUSIONS: ATF3 gene variants influence the risk of hypospadias. Its hormonal responsiveness may underlie this risk effect. But also other ATF3-dependent biological aspects, such as cell survival and death, response to stress stimuli, or the control of epithelial-mesenchymal interactions, may be of importance. PMID- 18426834 TI - Letrozole once a week normalizes serum testosterone in obesity-related male hypogonadism. AB - OBJECTIVE: Isolated hypogonadotropic hypogonadism (IHH) is frequently observed in severely obese men, probably as a result of increased estradiol (E(2)) production and E(2)-mediated negative feedback on pituitary LH secretion. Aromatase inhibitors can reverse this process. This study evaluates whether letrozole once a week can normalize serum testosterone in severely obese men and maintain its long term effect. DESIGN: Open, uncontrolled 6-month pilot study in 12 severely obese men (body mass index>35.0 kg/m(2)) with obesity-related IHH and free testosterone levels <225 pmol/l, treated with 2.5 mg letrozole once a week for 6 months. RESULTS: Six weeks of treatment reduced total E(2) from 123+/-11 to 58+/ 7 pmol/l (P<0.001, mean+/-s.e.m.), and increased serum LH from 4.4+/-0.6 to 11.1+/-1.5 U/l (P<0.001). Total testosterone rose from 5.9+/-0.5 to 19.6+/-1.4 nmol/l (P<0.001), and free testosterone from 163+/-13 to 604+/-50 pmol/l (P<0.001). Total testosterone rose to within the normal range in all subjects, whereas free testosterone rose to supraphysiological levels in 7 out of 12 men. The testosterone and E(2) levels were stable throughout the week and during the 6 month treatment period. CONCLUSION: Letrozole 2.5 mg once a week produced a sustained normalization of serum total testosterone in obese men with IHH. However, free testosterone frequently rose to supraphysiological levels. Therefore, a starting dose <2.5 mg once a week is recommended. PMID- 18426835 TI - Elevated serum parathyroid hormone predicts impaired survival prognosis in a general aged population. AB - OBJECTIVE: Short-term studies on selected patients have indicated that elevated serum parathyroid hormone (PTH) is an independent risk factor of death. However, long-term data on unselected populations are lacking, thus far. In order to evaluate the predictive value of elevated serum PTH during the last years of life, random persons of age cohorts of 75, 80 and 85 years were followed for 17 years. DESIGN: A prospective cohort study. METHODS: Subjects (n=567) were investigated for calcaemic status including serum intact PTH, serum total calcium (CaT) and ionized calcium (Ca(2+)). Thorough clinical examinations included an assessment of co-morbidity. Mortality data were collected from National Census Records. RESULTS: Up to 93% of the subjects died within the follow-up. In contrast to Ca(2+) levels, high serum PTH (> or =63 ng/l, IV quartile cut point) was associated with significant over-mortality (HR=1.56, 95% CI: 1.29-1.88) and a 2.3-year reduction of median life expectancy. After controlling for age, gender, co-morbidity and creatinine, the prognostic impact of elevated serum PTH was still significant (HR=1.24, 95% CI: 1.01-1.53). The tendency for over-mortality was consistent in both genders, in all age groups as well as in subjects with varying co-morbidity, renal function, body mass index categories and Ca(2+) levels. CONCLUSIONS: Elevated serum PTH level is an independent predictor of impaired long-term survival prognosis in unselected aged population. Serum Ca(2+) did not emerge as a significant prognostic indicator. The long-term prognostic impact of vitamin D deficiency, the most common cause of elevated PTH levels in the elderly, remains to be investigated. PMID- 18426836 TI - Variable number of tandem repeats polymorphism in parathyroid hormone-related protein as predictor of peak bone mass in young healthy Finnish males. AB - OBJECTIVE: Mice with osteoblast-specific deletion of parathyroid hormone-related protein (PTHrP) exhibit impaired recruitment and increased apoptosis of osteogenic cells resulting in decreased bone formation and premature osteoporosis. The PTHrP levels within the bone microenvironment are therefore critical in influencing bone mass acquisition. Whether this is applicable in humans has not been established. Here, we studied the association of a variable number of tandem repeats (VNTR) polymorphism in PTHrP with peak bone mass. METHODS: Enrolled in the study were 234 young Finnish males, with median age of 19.6 years (range 18.3-20.6 years). Lifestyle factors, serum bone markers, osteodensitometric measurements (lumbar spine and hip) and calcaneal quantitative ultrasound readings were obtained. The PTHrP VNTR length was determined by the PCR amplification of genomic DNA extracted from peripheral blood and correlated to bone parameters by the multiple regression models. RESULTS: The presence of at least one 252 bp allele was associated with increased lumbar spine bone mineral density (BMD; P<0.0034), broadband ultrasound attenuation (BUA; P<0.0012) and speed-of-sound (SOS; P<0.0023) measurements. The correlation with increased lumbar spine BMD (P=0.0008), BUA (P=0.005) and SOS (P=0.001) was further strengthened by the pairing of the 252 bp allele with a 460 bp allele in comparison with those without any 252 bp allele. Electrophoretic mobility shift assays were used to illustrate the potential transcriptional functionality of the VNTR sequence. CONCLUSION: The results indicate that the PTHrP VNTR sequence likely modulates local PTHrP expression within the skeletal microenvironment and could serve as a diagnostic predictor of peak bone mass acquisition. PMID- 18426837 TI - Retinol, retinol-binding protein 4, abdominal fat mass, peak bone mineral density, and markers of bone metabolism in men: the Northern Osteoporosis and Obesity (NO2) Study. AB - CONTEXT: The association between retinol and bone mineral density (BMD) in males after puberty has not been fully investigated previously. OBJECTIVE: To investigate the association between retinol, retinol-binding protein-4 (RBP-4), BMD (g/cm(2)), abdominal fat mass, and markers of bone metabolism in young men. DESIGN: Longitudinal study. PARTICIPANTS: Seventy-eight healthy males with a mean age of 22.6+/-0.7 years at baseline. A follow-up was conducted in 73 of the participants 2.0+/-0.4 years later. MAIN OUTCOME MEASURES: Associations between serum concentrations of retinol and RBP-4, and BMD of the total body, lumbar spine, and hip, serum concentrations of osteocalcin, and carboxy terminal telopeptide of type 1 collagen (CTX), were investigated. RESULTS: Both retinol and RBP-4 showed an inverse relationship with that of osteocalcin (r=-0.23 to 0.25, P<0.05). Levels of RBP-4 (r=0.26, P=0.02) and osteocalcin (r=-0.23, P=0.04) were also related to abdominal fat mass, and the relationship between RBP-4, retinol, and osteocalcin disappeared after adjusting for this influence of abdominal fat mass. Neither retinol nor RBP-4 concentrations were associated with BMD at any site, CTX as baseline, or changes in BMD during the 2-year follow-up period. Levels of RBP-4 showed a strong association with levels of retinol (r=0.61, P<0.001). CONCLUSION: We found a negative association between the bone formation marker osteocalcin with retinol and RBP-4. The association disappeared when adjusting for the influence of abdominal fat mass. Neither retinol nor RBP-4 were associated with peak BMD in young men. PMID- 18426838 TI - Lack of imatinib-induced thyroid dysfunction in a cohort of non-thyroidectomized patients. PMID- 18426839 TI - Screening for and prevention of type 2 diabetes. PMID- 18426840 TI - Different strategies for screening and prevention of type 2 diabetes in adults: cost effectiveness analysis. AB - OBJECTIVE: To compare four potential screening strategies, and subsequent interventions, for the prevention and treatment of type 2 diabetes: (a) screening for type 2 diabetes to enable early detection and treatment, (b) screening for type 2 diabetes and impaired glucose tolerance, intervening with lifestyle interventions in those with a diagnosis of impaired glucose tolerance to delay or prevent diabetes, (c) as for (b) but with pharmacological interventions, and (d) no screening. DESIGN: Cost effectiveness analysis based on development and evaluation of probabilistic, comprehensive economic decision analytic model, from screening to death. SETTING: A hypothetical population, aged 45 at time of screening, with above average risk of diabetes. DATA SOURCES: Published clinical trials and epidemiological studies retrieved from electronic bibliographic databases; supplementary data obtained from the Department of Health statistics for England and Wales, the screening those at risk (STAR) study, and the Leicester division of the ADDITION study. METHODS: A hybrid decision tree/Markov model was developed to simulate the long term effects of each screening strategy, in terms of both clinical and cost effectiveness outcomes. The base case model assumed a 50 year time horizon with discounting of both costs and benefits at 3.5%. Sensitivity analyses were carried out to investigate assumptions of the model and to identify which model inputs had most impact on the results. RESULTS: Estimated costs for each quality adjusted life year (QALY) gained (discounted at 3.5% a year for both costs and benefits) were pound14,150 (euro17 560; $27,860) for screening for type 2 diabetes, pound6242 for screening for diabetes and impaired glucose tolerance followed by lifestyle interventions, and pound7023 for screening for diabetes and impaired glucose tolerance followed by pharmacological interventions, all compared with no screening. At a willingness-to-pay threshold of pound20,000 the probability of the intervention being cost effective was 49%, 93%, and 85% for each of the active screening strategies respectively. CONCLUSIONS: Screening for type 2 diabetes and impaired glucose tolerance, with appropriate intervention for those with impaired glucose tolerance, in an above average risk population aged 45, seems to be cost effective. The cost effectiveness of a policy of screening for diabetes alone, which offered no intervention to those with impaired glucose tolerance, is still uncertain. PMID- 18426842 TI - How to improve surgical outcomes. PMID- 18426843 TI - Volume of procedures and risk of recurrence after repair of groin hernia: national register study. AB - OBJECTIVE: To determine whether the association between volume and outcome found in major surgery also holds true for a minor operation. DESIGN: Review of outcomes after hernia surgery in Sweden. SETTING: Surgical units registered with the Swedish hernia register, which in 2004 covered about 95% of all hernia operations in Sweden. PARTICIPANTS: 86,409 patients over 15 years, who underwent 96,601 unilateral or bilateral groin hernia repairs (94,077 inguinal and 2524 femoral) in 1996-2004 at the participating surgical units. MAIN OUTCOME MEASURE: Re-operation for recurrence. RESULTS: There was a significantly higher rate of re operation in surgeons who carried out 1-5 repairs a year than in surgeons who carried out more repairs. There was no association between outcome and further increases in volume. Although about half of surgeons in Sweden who repair hernias are low volume operators, they performed only 8.4% of all repairs. CONCLUSIONS: Sweden's numerous low volume hernia surgeons perform such a small proportion of all operations that the impact of their inferior results on the nationwide re operation rate is minimal. Volume indicates an approximate minimum value for the number of hernia repairs a surgeon should do each year but the outcome in surgeons who carry out more than that number disqualifies volume as an indicator of proficiency. PMID- 18426844 TI - Expanded safety study of Praneem polyherbal vaginal tablet among HIV-uninfected women in Pune, India: a phase II clinical trial report. AB - BACKGROUND: Praneem, a candidate vaginal microbicide with reported anti-HIV and contraceptive properties was evaluated for its long-term safety through a double blind, placebo-controlled, randomised phase II trial. METHODOLOGY: 142 women were screened and 100 HIV-uninfected, eligible women were enrolled, after obtaining written informed consents. Of these, 50 were randomly assigned to the study product arm and 50 to the placebo arm. Participants were requested to use the study product at least half an hour before each sexual act for 6 months. RESULTS: Participants in both treatment groups contributed 601 person-months of follow-up and 95% of the participants completed the last follow-up. Although transient genital discomfort was the main reported adverse experience, none of the study participants discontinued product use. Most of the adverse events were mild and there were no serious adverse events related to study product use and no HIV seroconversions. DISCUSSION: Praneem polyherbal tablet was safe for vaginal use up to 6 months with each act of sex among low-risk women. In light of the failure of Nonoxynol-9, cellulose sulphate and Carraguard in preventing HIV infections among women, additional preclinical evaluations should be undertaken urgently before moving into effectiveness studies. PMID- 18426845 TI - The cost-effectiveness of syndromic management for male sexually transmitted disease patients with urethral discharge symptoms and genital ulcer disease in Taiwan. AB - OBJECTIVE: To assess the cost-effectiveness of syndromic management for the treatment of sexually transmitted diseases (STD) in comparison with the strategies currently used in STD clinics in Taiwan. METHODS: Between July 2004 and June 2005, all male patients with genitourinary symptoms attending two hospitals were included in this study. Current clinical procedures (presumptive treatment and laboratory tests), aetiological diagnosis and syndromic management were compared in terms of diagnostic accuracy, treatment appropriateness, costs and effectiveness. RESULTS: 473 patients met the inclusion criteria and were enrolled in the study. 335 patients (71%) had urethral symptoms (discharge, dysuria or painful urination) and 138 (29%) had genital ulcers, sores and skin rashes. For the current approach, the sensitivity, specificity and positive predictive value (PPV) for the detection of chlamydial, gonococcal and combined forms of infection were 100%, 40.0% and 60.4%, respectively. In contrast, the sensitivity, specificity and PPV for the detection of syphilis were 100%, 86.7% and 70.2%, respectively. For syndromic management, the sensitivity, specificity and PPV detection of chlamydial, gonococcal and combined forms of infection were 85.0%, 40.0% and 56.4%, respectively. In contrast, the sensitivity, specificity and PPV for the detection of syphilis were 78.8%, 18.1% and 23.2%, respectively. The average cost of implementing a correct treatment using the current approach was US$54.27 and US$30.74 for urethritis and syphilis, respectively. For the aetiological approach, the average cost of implementing a correct treatment was US$32.83 and US$21.58 for urethritis and syphilis, respectively. For the syndromic approach, the average cost for a correct treatment was US$3.86 and US$14.30 for urethritis and syphilis, respectively. CONCLUSIONS: In this sample of patients attending STD clinics in Taiwan, syndromic management was found to be a more cost-effective protocol in terms of cost per treated STD patient compared with the current and aetiological protocols for STD. PMID- 18426846 TI - Ethics, industry, and outcomes. AB - Evidence-based medicine depends on research that is reliable, well conducted, and free of influence from interest groups (which are not confined to industry). There are many ways to influence the outcome of clinical research, and the history of influence in research related to tobacco is illuminating in this regard. Health care depends on industry, and if properly managed, the relationship between industry and medical academia can be symbiotic. Achieving positive outcomes from such relationships depends on ensuring the presence of the elements needed for reasonable independence on the part of investigators, and on understanding the ways in which influence can be exerted over the production and publication of evidence. Regulation cannot substitute for integrity, particularly the integrity of the investigators. Pushing the limits of ethics in research threatens to undermine not only the reputation of those conducting the research but also the standing of science itself. PMID- 18426847 TI - Early detection of an acute cerebral event during cardiopulmonary bypass using a bispectral index monitor. AB - A case in which the bispectral index monitor underwent an acute change that was most likely the earliest indicator of an intraoperative stroke during replacement of an aortic valve is discussed in this study. The timely response to this intraoperative event may have facilitated a better outcome by virtue of more rapid interventions to limit the severity of the injury. PMID- 18426848 TI - An estimate of the influence of the measurement procedure on patient and phantom doses in breast imaging. AB - This work is aimed to study the variability of dosimetry results owing to various measurement methodologies for breast dosimetry. This is performed in the frame of the development of a national protocol for breast dosimetry. Doses for standard phantom and group of patients were calculated for two mammography systems from the tube output measured with a calibrated ionisation chamber. The backscatter from the phantom under the chamber contributes to an increase in dosimeter readings of approximately 0.8-1.5%, whereas the proximity of the compression plate to the chamber causes increase in the measured air kerma value by 6.5-7%. High value layer (HVL) measured with solid-state detector without corrections for energy dependence was 17% higher than the one measured with ionisation chamber, which causes corresponding overestimation of average glandular dose (AGD). The use of conversion factors based on typical but not measured HVL values leads to 3.5-5.6% overestimation of AGD. Although the sources of uncertainty were taken into account, the difference between the phantom and patient doses was 24%. Some practical recommendations to be included in the national dosimetry protocol are summarised. PMID- 18426849 TI - Double-dosimetry algorithm for workers in interventional radiology. AB - Based on double-dosemeter readings, a conservative effective dose (E) estimation algorithm for lead apron workers in interventional radiology is proposed. Typical radiation conditions for various exposure geometries were simulated using the MCNPX 2.4.0 code. The simulation model consisted of an X-ray source and image intensifier, a patient phantom and a voxelised staff member phantom with lead apron. The effective staff dose and dosemeter readings for several positions of the worker were calculated. The effective dose to a physician, positioned in close proximity to the primary beam, can be estimated within a 10% underestimation margin by E = 1.64 H(p)(10)thorax,under + 0.075 H(p)(10)neck,over. The dose to the eye lens can be estimated by a dosemeter reading at collar level (R2 = 0.98). PMID- 18426850 TI - Running: the risk of coronary events : Prevalence and prognostic relevance of coronary atherosclerosis in marathon runners. AB - AIMS: To quantify the prevalence of coronary artery calcification (CAC) in relation to cardiovascular risk factors in marathon runners, and to study its role for myocardial damage and coronary events. METHODS AND RESULTS: In 108 apparently healthy male marathon runners aged >or=50 years, with >or=5 marathon competitions during the previous three years, the running history, Framingham risk score (FRS), CAC, and presence of myocardial late gadolinium enhancement (LGE) were measured. Control groups were matched by age (8:1) and FRS (2:1) from the Heinz Nixdorf Recall Study. The FRS in marathon runners was lower than in age matched controls (7 vs. 11%, P < 0.0001). However, the CAC distribution was similar in marathon runners and age-matched controls (median CAC: 36 vs. 38, P = 0.36) and higher in marathon runners than in FRS-matched controls (median CAC: 36 vs. 12, P = 0.02). CAC percentile values and number of marathons independently predicted the presence of LGE (prevalence = 12%) (P = 0.02 for both). During follow-up after 21.3 +/- 2.8 months, four runners with CAC >or= 100 experienced coronary events. Event-free survival was inversely related to CAC burden (P = 0.018). CONCLUSION: Conventional cardiovascular risk stratification underestimates the CAC burden in presumably healthy marathon runners. As CAC burden and frequent marathon running seem to correlate with subclinical myocardial damage, an increased awareness of a potentially higher than anticipated coronary risk is warranted. PMID- 18426851 TI - Early intratracheal instillation of budesonide using surfactant as a vehicle to prevent chronic lung disease in preterm infants: a pilot study. AB - OBJECTIVE: Budesonide is an inhaled steroid with a strong topical effect but with minimal systemic effects; it has been effectively delivered to animal lungs using surfactant as a vehicle. The purposes of this study were to determine whether early intratracheal instillation of budesonide using surfactant as a vehicle would improve pulmonary status, reduce mortality, and reduce chronic lung disease morbidity. PATIENTS AND METHODS: We conducted a prospective, randomized blind trial in 116 very low birth weight infants (< 1500 g) who had severe radiographic respiratory distress syndrome and required mechanical ventilation with fraction of inspired oxygen > or = 0.6 shortly after birth: 60 were in the treated group (intratracheal instillation of a mixture of 0.25 mg/kg of budesonide and 100.00 mg/kg of survanta, every 8 hours) and 56 were in the control group (100 mg/kg of survanta only, every 8 hours). The end point assessment was the number of infants who would die or develop chronic lung disease at 36 weeks' postconceptional age. RESULTS: Infants in the treatment group required significantly lower mean airway pressure on day 1 and day 3 and had significantly lower oxygen index and PCO(2) during the first 3 days than infants in the control group. More infants were extubated in the treatment group than controls at 1 and 2 weeks. The combined outcome of deaths or chronic lung disease was significantly lower in the treatment group than in the control group (19 of 60 vs 34 of 56). No clinically significant adverse effects were observed during the study. CONCLUSIONS: This pilot study indicated that early postnatal intratracheal instillation of budesonide using surfactant as vehicle significantly improved the combined outcome of death or chronic lung disease in small premature infants without causing immediate adverse effects. The results are encouraging, and a large sample multicenter trial is warranted. PMID- 18426852 TI - Predictors of retinochoroiditis in children with congenital toxoplasmosis: European, prospective cohort study. AB - OBJECTIVE: By school age, 20% of children infected with congenital toxoplasmosis will have > or = 1 retinochoroidal lesion. We determined which children are most at risk and whether prenatal treatment reduces the risk of retinochoroiditis to help clinicians decide about treatment and follow-up. PATIENTS AND METHODS: We prospectively studied a cohort of children with congenital toxoplasmosis identified by prenatal or neonatal screening in 6 European countries. We determined the effects of prenatal treatment and prognostic markers soon after birth on the age at first detection of retinochoroiditis. RESULTS: Of 281 children with congenital toxoplasmosis, 50 developed ocular disease, and 17 had recurrent retinochoroiditis during a median follow-up of 4.1 years. Prenatal treatment had no significant effect on the age at first or subsequent lesions. Delayed start of postnatal treatment did not increase retinochoroiditis, but the analysis lacked power. Older gestational age at maternal seroconversion was weakly associated with a reduced risk of retinochoroiditis. The presence of nonocular clinical manifestations of congenital toxoplasmosis at birth strongly predicted retinochoroiditis. For 92% (230 of 249) of children with no retinochoroiditis detected before 4 months of age, the probability of retinochoroiditis by 4 years was low, whether clinical manifestations were present or not 8.0%. CONCLUSIONS: Prenatal treatment did not significantly reduce the risk of retinochoroiditis in this European cohort. If children have no retinochoroiditis in early infancy, the low risk of subsequent ocular disease may not justify postnatal treatment and repeated ophthalmic assessments during childhood. Controlled trials are needed to address the lack of evidence for the effectiveness of postnatal treatment. PMID- 18426853 TI - Erythroid apoptosis in idiopathic neonatal jaundice. AB - OBJECTIVES: The objectives of this study were to evaluate the contribution of erythroid apoptosis to neonatal idiopathic pathologic jaundice and to determine whether a measurement of the erythroid apoptosis value at birth could predict the development of hyperbilirubinemia during the first 15 days of life. PATIENTS AND METHODS: Three groups were defined: group 1 (n = 101), healthy newborns whose erythroid apoptosis value and serum total bilirubin levels were detected from birth to day 15; group 2 (n = 24), newborns who were hospitalized for jaundice (serum total bilirubin level: > 12.9 mg/dL) without any identifiable pathologic cause; and group 3 (control group, n = 24), healthy newborns whose serum total bilirubin levels were < or = 12.9 mg/dL. Erythroid apoptosis value was assessed by flow cytometry using an annexin-V fluorescein isothiocyanate kit. RESULTS: In group 1, there was no correlation between the erythroid apoptosis value and serum total bilirubin levels obtained at birth and at the fourth and 15th days of life; the erythrocyte apoptosis value obtained at birth was not significantly different between the neonates whose serum total bilirubin levels were > 12.9 and < or = 12.9 mg/dL and who had prolonged and nonprolonged jaundice during follow-up. The erythroid apoptosis value differed significantly between the newborns in groups 2 and 3. There was no significant correlation between the erythroid apoptosis value and serum total bilirubin levels of the infants in groups 2 and 3. CONCLUSIONS: The erythroid apoptosis value obtained at birth could not predict the development of hyperbilirubinemia in neonates, but it was increased significantly in jaundiced neonates whose serum total bilirubin levels were > 12.9 mg/dL. In these infants, increase in the erythroid apoptosis value may be a result of the toxic effect of bilirubin or of a protective mechanism of neonates to increase heme turnover and bilirubin production to diminish oxidative stress. PMID- 18426854 TI - High resolution computed tomography imaging for individualized allograft replacement of an infected Dacron aortic arch prosthesis. AB - We report on the case of a young woman with a history of intravenous drug abuse and the development of an infection of a prosthetic supracoronary-, total arch and partial descending aortic allograft prosthesis following acute aortic dissection type Stanford-A two years previously. For surgical treatment we implanted an allograft prosthesis obtained from a local tissue bank. Particular variations in this setting and in comparison to conventional surgical planning were that high resolution computed tomography imaging was applied to determine and subsequently order the optimal allograft prosthesis in this individual patient and anatomical situation. PMID- 18426855 TI - Comparison of non-parametric confidence intervals for the area under the ROC curve of a continuous-scale diagnostic test. AB - The accuracy of a diagnostic test with continuous-scale results is of high importance in clinical medicine. It is often summarised by the area under the ROC curve (AUC). In this article, we discuss and compare nine non-parametric confidence intervals of the AUC for a continuous-scale diagnostic test. Simulation studies are conducted to evaluate the relative performance of the confidence intervals for the AUC in terms of coverage probability and average interval length. A real example is used to illustrate the application of the recommended methods. PMID- 18426856 TI - Selective killing of Smad4-negative tumor cells via a designed repressor strategy. AB - Smad4 is a key tumor suppressor that is frequently deleted or inactive in pancreatic and colon tumors. In this report, we describe an approach for attaining selective killing of Smad4-deficient tumor cells. Using a vector system involving a designed repressor with zinc finger binding domains and the herpes simplex virus thymidine kinase (HSV-TK) "suicide gene," we demonstrate Smad4 responsive regulation of HSV-TK expression and consequent altered susceptibility to the prodrug ganciclovir (GCV). In pancreatic tumor cell lines stably transfected with the vector system, a robust differential of HSV-TK expression and GCV toxicity was attained depending on the presence or absence of cotransfected Smad4. In matched colon tumor cell lines lacking Smad4 or expressing physiological levels of Smad4, an adenoviral version of the vector system attained a significant degree of preferential killing of Smad4-negative tumor cells in response to GCV. These findings demonstrate the possibility of achieving selective killing of pancreatic and colon cells depending on their Smad4 status. PMID- 18426857 TI - Nitric oxide donor, (+/-)-S-nitroso-N-acetylpenicillamine, stabilizes transactive hypoxia-inducible factor-1alpha by inhibiting von Hippel-Lindau recruitment and asparagine hydroxylation. AB - We have confirmed that the NO donor (+/-)-S-nitroso-N-acetylpenicillamine (SNAP) stabilizes the transactive form of hypoxia-inducible factor-1alpha (HIF-1alpha), leading to the induction of HIF-1alpha target genes such as vascular endothelial growth factor and carbonic anhydrase 9. Activation of HIF-1alpha should require inhibition of the dual system that keeps it inactive. One is ubiquitination, which is triggered by hydroxylation of HIF-1alpha-proline and the subsequent binding of E3 ubiquitin ligase, the von Hippel Lindau (VHL) protein. The other is hydroxylation of HIF-1alpha-asparagine, which reduces the affinity of HIF-1alpha for its coactivator, cAMP responsive element binding protein/p300. We examined the effects of the NO donor SNAP on proline and asparagine hydroxylation of HIF 1alpha peptides by measuring the activities of the corresponding enzymes, HIF 1alpha-specific proline hydroxylase 2 (PHD2) and the HIF-1alpha-specific asparagine hydroxylase, designated factor inhibiting HIF-1alpha (FIH-1), respectively. We found that the SNAP did not prevent PHD2 from hydroxylating the proline of HIF-1alpha. Instead, it blocked the interaction between VHL and the proline-hydroxylated HIF-1alpha, but only when the reducing agents Fe(II) and vitamin C were limiting. The fact that the absence of cysteine 520 of HIF-1alpha abolishes its responsiveness to SNAP suggests that this residue mediates the inhibition by SNAP of the interaction between VHL and HIF-1alpha, presumably by S nitrosylation of HIF-1alpha. Un-like PHD2, asparagine hydroxylation by FIH-1 was directly inhibited by SNAP, but again only when reducing agents were limiting. Substitution of cysteine 800 of HIF-1alpha with alanine failed to reverse the inhibitory effects of SNAP on asparagine hydroxylation, implying that FIH-1, not its substrate HIF-1alpha, is inhibited by SNAP. PMID- 18426858 TI - Baicalein induces functional hypoxia-inducible factor-1alpha and angiogenesis. AB - Targeting the oxygen-sensing mechanisms of the hypoxiainducible factor (HIF) pathway provides pharmacological ways of manipulating the HIF response. Because HIF-1alpha-specific prolyl-4 hydroxylases (PHDs) prime degradation of HIF-1alpha, we have made an effort to find a small molecule capable of up-regulating the HIF pathway by inhibiting prolyl hydroxylation. Through an in vitro high-throughput screen, we have discovered a PHD2 inhibitor baicalein, which is also found to abrogate asparaginyl hydroxylation of HIF-1alpha. Such inhibitory effects are reversed by the addition of excess 2-oxoglutarate and iron(II), suggesting the involvement of baicalein's binding at the enzyme active sites, which has also been corroborated by spectroscopic binding assays between baicalein and enzyme. In addition, baicalein suppresses ubiquitination of HIF-1alpha, which works in concert with the inhibition of the HIF-specific hydroxylases to increase the HIF 1alpha content, leading to induction of HIF-1-mediated reporter gene activity and target gene transcription in tissue culture cells, whereas it induces HIF independent activation of other genes. Furthermore, in vivo organ models based on the chick chorioallantoic membrane assay demonstrate that baicalein promotes new blood vessel formation. Together, our results indicate that baicalein possesses a proangiogenic potential and thus might have the therapeutic utility in the treatment of ischemic diseases. PMID- 18426859 TI - Association of 1,5-anhydroglucitol and 2-h postprandial blood glucose in type 2 diabetic patients. AB - OBJECTIVE: To assess the association of 1,5-anhydroglucitol (1,5-AG) with 2-h postprandial glucose values in type 2 diabetic patients followed over 12 months in an outpatient setting. RESEARCH DESIGN AND METHODS: In 55 patients, we examined self-measured postprandial blood glucose values for correlations with 1,5-AG values over prespecified preceding time periods (3 days, 1 week, and weekly up to 12 weeks). RESULTS: The correlation coefficients for postprandial glucose values were -0.34 (P < 0.05) for 3 days, -0.38 (P < 0.001) for 1 week, and -0.40 (P < 0.001) for 2 weeks preceding the measurement of 1,5-AG. Correlations declined for time periods >2 weeks before measurement of 1,5-AG. The correlation was lower with fasting/preprandial plasma glucose levels. There was no time dependency for the correlation between A1C and fasting or postprandial glucose. CONCLUSIONS: 1,5-AG best reflected the 2-h postprandial glucose values of the 2 previous weeks. PMID- 18426860 TI - Protein engineering strategies for sustained glucagon-like peptide-1 receptor dependent control of glucose homeostasis. AB - OBJECTIVE: We have developed a novel platform for display and delivery of bioactive peptides that links the biological properties of the peptide to the pharmacokinetic properties of an antibody. Peptides engineered in the MIMETIBODY platform have improved biochemical and biophysical properties that are quite distinct from those of Fc-fusion proteins. CNTO736 is a glucagon-like peptide 1 (GLP-1) receptor agonist engineered in our MIMETIBODY platform. It retains many activities of native GLP-1 yet has a significantly enhanced pharmacokinetic profile. Our goal was to develop a long-acting GLP-1 receptor agonist with sustained efficacy. RESEARCH DESIGN AND METHODS: In vitro and in vivo activity of CNTO736 was evaluated using a variety of rodent cell lines and diabetic animal models. RESULTS: Acute pharmacodynamic studies in diabetic rodents demonstrate that CNTO736 reduces fasting and postprandial glucose, decreases gastric emptying, and inhibits food intake in a GLP-1 receptor-specific manner. Reduction of food intake following CNTO736 dosing is coincident with detection of the molecule in the circumventricular organs of the brain and activation of c-fos in regions protected by the blood-brain barrier. Diabetic rodents dosed chronically with CNTO736 have lower fasting and postprandial glucose and reduced body weight. CONCLUSIONS: Taken together, our data demonstrate that CNTO736 produces a spectrum of GLP-1 receptor-dependent actions while exhibiting significantly improved pharmacokinetics relative to the native GLP-1 peptide. PMID- 18426861 TI - Association analysis of type 2 diabetes Loci in type 1 diabetes. AB - OBJECTIVE: To search for a possible association of type 1 diabetes with 10 validated type 2 diabetes loci, i.e., PPARG, KCNJ11, WFS1, HNF1B, IDE/HHEX, SLC30A8, CDKAL1, CDKN2A/B, IGF2BP2, and FTO/RPGRIP1L. RESEARCH DESIGN AND METHODS: Two European population samples were studied: 1) one case-control cohort of 514 type 1 diabetic subjects and 2,027 control subjects and 2) one family cohort of 483 complete type 1 diabetic case-parent trios (total 997 affected). A total of 13 tag single nucleotide polymorphisms (SNPs) from the 10 type 2 diabetes loci were analyzed for type 1 diabetes association. RESULTS: No association of type 1 diabetes was found with any of the 10 type 2 diabetes loci, and no age-at-onset effect was detected. By combined analysis using the Wellcome Trust Case-Control Consortium type 1 diabetes data, SNP rs1412829 in the CDKN2A/B locus bordered on significance (P = 0.039) (odds ratio 0.929 [95% CI 0.867 0.995]), which did not reach the statistical significance threshold adjusted for 13 tests (alpha = 0.00385). CONCLUSIONS: This study suggests that the type 2 diabetes loci do not play any obvious role in type 1 diabetes genetic susceptibility. The distinct molecular mechanisms of the two diseases highlighted the importance of differentiation diagnosis and different treatment principles. PMID- 18426862 TI - Haplotype structure of the ENPP1 Gene and Nominal Association of the K121Q missense single nucleotide polymorphism with glycemic traits in the Framingham Heart Study. AB - OBJECTIVE: A recent meta-analysis demonstrated a nominal association of the ectonucleotide pyrophosphatase phosphodiesterase 1 (ENPP1) K-->Q missense single nucleotide polymorphism (SNP) at position 121 with type 2 diabetes. We set out to confirm the association of ENPP1 K121Q with hyperglycemia, expand this association to insulin resistance traits, and determine whether the association stems from K121Q or another variant in linkage disequilibrium with it. RESEARCH DESIGN AND METHODS: We characterized the haplotype structure of ENPP1 and selected 39 tag SNPs that captured 96% of common variation in the region (minor allele frequency > or =5%) with an r(2) value > or =0.80. We genotyped the SNPs in 2,511 Framingham Heart Study participants and used age- and sex-adjusted linear mixed effects (LME) models to test for association with quantitative metabolic traits. We also examined whether interaction between K121Q and BMI affected glycemic trait levels. RESULTS: The Q allele of K121Q (rs1044498) was associated with increased fasting plasma glucose (FPG), A1C, fasting insulin, and insulin resistance by homeostasis model assessment (HOMA-IR; all P = 0.01-0.006). Two noncoding SNPs (rs7775386 and rs7773477) demonstrated similar associations, but LME models indicated that their effects were not independent from K121Q. We found no association of K121Q with obesity, but interaction models suggested that the effect of the Q allele on FPG and HOMA-IR was stronger in those with a higher BMI (P = 0.008 and 0.01 for interaction, respectively). CONCLUSIONS: The Q allele of ENPP1 K121Q is associated with hyperglycemia and insulin resistance in whites. We found an adiposity-SNP interaction, with a stronger association of K121Q with diabetes-related quantitative traits in people with a higher BMI. PMID- 18426863 TI - Circulating retinol-binding protein-4 concentration might reflect insulin resistance-associated iron overload. AB - OBJECTIVES: The mechanisms behind the association between retinol-binding protein 4 (RBP4) and insulin resistance are not well understood. An interaction between iron and vitamin A status, of which RBP4 is a surrogate, has long been recognized. We hypothesized that iron-associated insulin resistance could be behind the impaired insulin action caused by RBP4. RESEARCH DESIGN AND METHODS: Serum ferritin and RBP4 concentration and insulin resistance were evaluated in a sample of middle-aged men (n = 132) and in a replication independent study. Serum RBP4 was also studied before and after iron depletion in patients with type 2 diabetes. Finally, the effect of iron on RBP4 release was evaluated in vitro in adipose tissue. RESULTS: A positive correlation between circulating RBP4 and log serum ferritin (r = 0.35 and r = 0.61, respectively; P < 0.0001) was observed in both independent studies. Serum RBP4 concentration was higher in men than women in parallel to increased ferritin levels. On multiple regression analyses to predict serum RBP4, log serum ferritin contributed significantly to RBP4 variance after controlling for BMI, age, and homeostasis model assessment value. Serum RBP4 concentration decreased after iron depletion in type 2 diabetic patients (percent mean difference -13.7 [95% CI -25.4 to -2.04]; P = 0.024). The iron donor lactoferrin led to increased dose-dependent adipose tissue release of RBP4 (2.4-fold, P = 0.005) and increased RBP4 expression, while apotransferrin and deferoxamine led to decreased RBP4 release. CONCLUSIONS: The relationship between circulating RBP4 and iron stores, both cross-sectional and after iron depletion, and in vitro findings suggest that iron could play a role in the RBP4-insulin resistance relationship. PMID- 18426864 TI - Blockade of alpha4 integrin signaling ameliorates the metabolic consequences of high-fat diet-induced obesity. AB - OBJECTIVE: Many prevalent diseases of advanced societies, such as obesity-induced type 2 diabetes, are linked to indolent mononuclear cell-dependent inflammation. We previously proposed that blockade of alpha4 integrin signaling can inhibit inflammation while limiting mechanism-based toxicities of loss of alpha4 function. Thus, we hypothesized that mice bearing an alpha4(Y991A) mutation, which blocks signaling, would be protected from development of high-fat diet induced insulin resistance. RESEARCH DESIGN AND METHODS: Six- to eight-week-old wild-type and alpha4(Y991A) C57Bl/6 male mice were placed on either a high-fat diet that derived 60% calories from lipids or a chow diet. Metabolic testing was performed after 16-22 weeks of diet. RESULTS: Alpha4(Y991A) mice were protected from development of high-fat diet-induced insulin resistance. This protection was conferred on wild-type mice by alpha4(Y991A) bone marrow transplantation. In the reverse experiment, wild-type bone marrow renders high-fat diet-fed alpha4(Y991A) acceptor animals insulin resistant. Furthermore, fat-fed alpha4(Y991A) mice showed a dramatic reduction of monocyte/macrophages in adipose tissue. This reduction was due to reduced monocyte/macrophage migration rather than reduced monocyte chemoattractant protein-1 production. CONCLUSIONS: Alpha4 integrins contribute to the development of HFD-induced insulin resistance by mediating the trafficking of monocytes into adipose tissue; hence, blockade of alpha4 integrin signaling can prevent the development of obesity-induced insulin resistance. PMID- 18426865 TI - Activation of estrogen receptor is crucial for resveratrol-stimulating muscular glucose uptake via both insulin-dependent and -independent pathways. AB - OBJECTIVE: Estradiol (E(2)) is known to modulate insulin sensitivity and, consequently, glucose homeostasis. Resveratrol (RSV), an agonist of estrogen receptor (ER), has exerted antihyperglycemic effects in streptozotocin-induced type 1 diabetic rats in our previous study and was also shown to improve insulin resistance in other reports. However, it remains unknown whether activation of ER is involved in the metabolic effects of RSV via insulin-dependent and independent mechanisms. RESEARCH DESIGN AND METHODS: Male Sprague-Dawley rats were given a high cholesterol-fructose (HCF) diet for 15 weeks and were treated with RSV for either 15 days or 15 weeks. RESULTS: Here, we show that RSV shifts the metabolic characteristics of rats on an HCF diet toward those of rats on a standard diet. RSV treatment increased insulin-stimulated whole-body glucose uptake and steady-state glucose uptake of soleus muscle and liver in HCF-fed rats as well as enhanced membrane trafficking activity of GLUT4 and increased phosphorylation of insulin receptor in insulin-resistant soleus muscles. Interestingly, the phosphorylated ER level in insulin-resistant soleus muscle was significantly elevated in rats with RSV treatment in both basal and euglycemic hyperinsulinemic conditions. RSV exerted an insulin-like stimulatory effect on isolated soleus muscle, epididymal fat and hepatic tissue, and C2C12 myotubes. The RSV-stimulated glucose uptake in C2C12 myotubes was dependent on extracellular signal-related kinase/p38 (early phase, 1 h) and p38/phosphoinositide 3-kinase (late phase, 14 h) activation. Inhibition of ER abrogated RSV-induced glucose uptake in both early and late phases. CONCLUSIONS: Collectively, these results indicate that ER is a key regulator in RSV stimulating insulin-dependent and -independent glucose uptake, which might account for the protective effects of RSV on diet-induced insulin resistance syndrome. PMID- 18426866 TI - Association of FTO with obesity-related traits in the Cebu Longitudinal Health and Nutrition Survey (CLHNS) Cohort. AB - OBJECTIVE: The underlying genetic component of obesity-related traits is not well understood, and there is limited evidence to support genetic association shared across multiple studies, populations, and environmental contexts. The present study investigated the association between candidate variants and obesity-related traits in a sample of 1,886 adult Filipino women from the Cebu Longitudinal Health and Nutrition Survey (CLHNS) cohort. RESEARCH DESIGN AND METHODS: We selected and genotyped 19 single nucleotide polymorphisms in 10 genes (ADRB2, ADRB3, FTO, GNB3, INSIG2, LEPR, PPARG, TNF, UCP2, and UCP3) that had been previously reported to be associated with an obesity-related quantitative trait. RESULTS: We observed evidence for association of the A allele of rs9939609 (FTO intron 1) with increased BMI (P = 0.0072 before multiple test correction), baseline BMI (P = 0.0015), longitudinal BMI based on eight surveys from 1983 to 2005 (P = 0.000029), waist circumference (P = 0.0094), and weight (P = 0.021). The increase in average BMI was approximately 0.4 for each additional A allele. We also observed association of the ADRB3 Trp64Arg variant with BMI, waist circumference, percent body fat, weight, fat mass, arm fat area, and arm muscle area (P < 0.05), although the direction of effect is inconsistent with the majority of previous reports. CONCLUSIONS: Our study confirms that FTO is a common obesity susceptibility gene in Filipinos, with an effect size similar to that seen in samples of European origin. PMID- 18426867 TI - Population tobacco control interventions and their effects on social inequalities in smoking: systematic review. AB - OBJECTIVE: To assess the effects of population tobacco control interventions on social inequalities in smoking. DATA SOURCES: Medical, nursing, psychological, social science and grey literature databases, bibliographies, hand-searches and contact with authors. STUDY SELECTION: Studies were included (n = 84) if they reported the effects of any population-level tobacco control intervention on smoking behaviour or attitudes in individuals or groups with different demographic or socioeconomic characteristics. DATA EXTRACTION: Data extraction and quality assessment for each study were conducted by one reviewer and checked by a second. DATA SYNTHESIS: Data were synthesised using graphical ("harvest plot") and narrative methods. No strong evidence of differential effects was found for smoking restrictions in workplaces and public places, although those in higher occupational groups may be more likely to change their attitudes or behaviour. Smoking restrictions in schools may be more effective in girls. Restrictions on sales to minors may be more effective in girls and younger children. Increasing the price of tobacco products may be more effective in reducing smoking among lower-income adults and those in manual occupations, although there was also some evidence to suggest that adults with higher levels of education may be more price-sensitive. Young people aged under 25 are also affected by price increases, with some evidence that boys and non-white young people may be more sensitive to price. CONCLUSIONS: Population-level tobacco control interventions have the potential to benefit more disadvantaged groups and thereby contribute to reducing health inequalities. PMID- 18426868 TI - What happened to smokers' beliefs about light cigarettes when "light/mild" brand descriptors were banned in the UK? Findings from the International Tobacco Control (ITC) Four Country Survey. AB - AIM: This paper examines how beliefs of smokers in the UK were affected by the removal of "light" and "mild" brand descriptors, which came into effect on 30 September 2003 for Member States of the European Union (EU). PARTICIPANTS: The data come from the first four waves (2002-2005) of the International Tobacco Control Policy Evaluation (ITC) Four-Country Survey, an annual cohort telephone survey of adult smokers in Canada, USA, UK and Australia (15 450 individual cases). DESIGN: The UK ban on misleading descriptors occurred around the second wave of data collection in the ITC survey, permitting us to compare beliefs about light cigarettes among adult smokers in the UK before and after the ban, with beliefs in the three other ITC countries unaffected by the ban. RESULTS: There was a substantial decline in reported beliefs about the benefits of light cigarettes in the UK following the policy change and an associated public information campaign, but by 2005 (ie, wave 4), these beliefs rebounded slightly and the change in beliefs was no greater than in the USA, where there was no policy change. CONCLUSIONS: The findings reveal that high levels of misperceptions about light cigarettes existed among smokers in all four countries before and after the EU ban took effect. We cannot conclude that the policy of removing some aspects of misleading labels has been effective in changing beliefs about light cigarettes. Efforts to correct decades of consumer misperceptions about light cigarettes must extend beyond simply removing "light" and "mild" brand descriptors. PMID- 18426869 TI - Serendipitous discovery of an immunoglobulin-binding autotransporter in Bordetella species. AB - We describe the serendipitous discovery of BatB, a classical-type Bordetella autotransporter (AT) protein with an approximately 180-kDa passenger domain that remains noncovalently associated with the outer membrane. Like genes encoding all characterized protein virulence factors in Bordetella species, batB transcription is positively regulated by the master virulence regulatory system BvgAS. BatB is predicted to share similarity with immunoglobulin A (IgA) proteases, and we showed that BatB binds Ig in vitro. In vivo, a Bordetella bronchiseptica DeltabatB mutant was unable to overcome innate immune defenses and was cleared from the lower respiratory tracts of mice more rapidly than wild-type B. bronchiseptica. This defect was abrogated in SCID mice, suggesting that BatB functions to resist clearance during the first week postinoculation in a manner dependent on B- and T-cell-mediated activities. Taken together with the previous demonstration that polymorphonuclear neutrophils (PMN) are critical for the control of B. bronchiseptica in mice, our data support the hypothesis that BatB prevents nonspecific antibodies from facilitating PMN-mediated clearance during the first few days postinoculation. Neither of the strictly human-adapted Bordetella subspecies produces a fully functional BatB protein; nucleotide differences within the putative promoter region prevent batB transcription in Bordetella pertussis, and although expressed, the batB gene of human-derived Bordetella parapertussis (B. parapertussis(hu)) contains a large in-frame deletion relative to batB of B. bronchiseptica. Taken together, our data suggest that BatB played an important role in the evolution of virulence and host specificity among the mammalian-adapted bordetellae. PMID- 18426870 TI - Characterization of the Ers regulon of Enterococcus faecalis. AB - Ers has been qualified as the PrfA-like transcriptional regulator of Enterococcus faecalis. In a previous study we reported that Ers is important for the survival within macrophages of this opportunist pathogenic bacterium. In the present work we have used proteomic and microarray expression profiling of E. faecalis JH2-2 and an ers-deleted mutant (Delta ers mutant) strains to define the Ers regulon. In addition to EF_0082 (encoding a putative facilitator family transporter), already known to be under Ers regulation, three genes or operons displayed a significant decrease (confirmed by reverse transcription quantitative PCR) in expression in the Delta ers mutant. The first locus corresponds to three genes: arcA, arcB, and arcC1 (arcABC). These genes are members of the ADI operon, encoding enzymes of the arginine deiminase system. The second is the EF_1459 gene, which encodes a hypothetical protein and is located within a putative phage genetic element. Lastly, Ef_3319 is annotated as the alpha subunit of the citrate lyase encoded by citF. citF is a member of a putative 12-gene operon involved in citrate catabolism. Moreover, the promoter sequence, similar to the "PrfA box" and found in the promoter regions of ers and EF_0082, has been shown to be included in the DNA segment recognized by Ers. Phenotypic analysis of the Delta ers mutant strain revealed a growth defect when cultured with arginine or citrate as the energy source; this was not seen for the wild type. As expected, similar results were obtained with mutants in which arcA and citF were inactivated. In addition, in the mouse peritonitis model of virulence, the Delta ers mutant appeared significantly less lethal than the JH2-2 wild-type strain. Taken together, these results indicate that the regulator Ers has a pleiotropic effect, especially in the cellular metabolism and virulence of E. faecalis. PMID- 18426871 TI - Francisella tularensis invasion of lung epithelial cells. AB - Francisella tularensis, a gram-negative facultative intracellular bacterial pathogen, causes disseminating infections in humans and other mammalian hosts. Macrophages and other monocytes have long been considered the primary site of F. tularensis replication in infected animals. However, recently it was reported that F. tularensis also invades and replicates within alveolar epithelial cells following inhalation in a mouse model of tularemia. TC-1 cells, a mouse lung epithelial cell line, were used to study the process of F. tularensis invasion and intracellular trafficking within nonphagocytic cells. Live and paraformaldehyde-fixed F. tularensis live vaccine strain organisms associated with, and were internalized by, TC-1 cells at similar frequencies and with indistinguishable differences in kinetics. Inhibitors of microfilament and microtubule activity resulted in significantly decreased F. tularensis invasion, as did inhibitors of phosphatidylinositol 3-kinase and tyrosine kinase activity. Collectively, these results suggest that F. tularensis epithelial cell invasion is mediated by a preformed ligand on the bacterial surface and driven entirely by host cell processes. Once internalized, F. tularensis-containing endosomes associated with early endosome antigen 1 (EEA1) followed by lysosome-associated membrane protein 1 (LAMP-1), with peak coassociation frequencies occurring at 30 and 120 min postinoculation, respectively. By 2 h postinoculation, 70.0% (+/- 5.5%) of intracellular bacteria were accessible to antibody delivered to the cytoplasm, indicating vacuolar breakdown and escape into the cytoplasm. PMID- 18426872 TI - Deficiency of indoleamine 2,3-dioxygenase enhances commensal-induced antibody responses and protects against Citrobacter rodentium-induced colitis. AB - Indoleamine 2,3-dioxygenase (IDO) is a negative regulator of lymphocyte responses that is expressed predominantly in macrophages and dendritic cells. We detected it at high levels in the small intestine and mesenteric lymph node of young adult mice, suggesting a role in intestinal immunity. Consistent with this idea, we found that IDO-deficient mice had elevated baseline levels of immunoglobulin A (IgA) and IgG in the serum and increased IgA in intestinal secretions. These abnormalities were corrected by a course of broad-spectrum oral antibiotics started at weaning, indicating that they were dependent on the intestinal microbiota. Kynurenine and picolinic acid, two IDO-generated metabolites of tryptophan, were able to inhibit lipopolysaccharide-induced antibody production by splenocytes in vitro, and kynurenine also induced B-cell apoptosis, findings that provide an explanation for the elevated Ig levels in animals lacking IDO. The intestinal secretions of IDO-deficient mice had elevated levels of IgA antibodies that cross-reacted with the gram-negative enteric bacterial pathogen Citrobacter rodentium. In keeping with the functional importance of this natural secretory IgA, the mutant animals were more resistant to intestinal colonization by Citrobacter, developed lower levels of serum Citrobacter-specific IgM and IgG antibodies following oral infection, and had significantly attenuated Citrobacter induced colitis. Our observations point to an important role for IDO in the regulation of immunity to the gut commensal microbiota that has a significant impact on the response to intestinal pathogens. PMID- 18426873 TI - Late endocytic multivesicular bodies intersect the chlamydial inclusion in the absence of CD63. AB - Chlamydiae are obligate intracellular bacterial pathogens that replicate solely within a membrane-bound vacuole termed an inclusion. Within the confines of the inclusion, the replicating bacteria acquire amino acids, nucleotides, and other precursors from the host cell. Trafficking from CD63-positive multivesicular bodies to the inclusion was previously identified as a novel interaction that provided essential precursors for the maintenance of a productive intracellular infection. The present study analyzes the direct delivery of resident protein and lipid constituents of multivesicular bodies to the intracellular chlamydiae. The manipulation of this trafficking pathway with an inhibitor of multivesicular body transport and the delivery of exogenous antibodies altered protein and cholesterol acquisition and delayed the maturation of the chlamydial inclusion. Although inhibitor studies and ultrastructural analyses confirmed a novel interaction between CD63-positive multivesicular bodies and the intracellular chlamydiae, neutralization with small interfering RNAs and anti-CD63 Fab fragments revealed that CD63 itself was not required for this association. These studies confirm CD63 as a constituent in multivesicular body-to-inclusion transport; however, other requisite components of these host cell compartments must control the delivery of key nutrients that are essential to intracellular bacterial development. PMID- 18426874 TI - Expression of Staphylococcus epidermidis SdrG increases following exposure to an in vivo environment. AB - SdrG is a surface-associated fibrinogen binding protein present in most strains of Staphylococcus epidermidis. Surface expression of SdrG was not detected by flow cytometry or immunofluorescence microscopy on S. epidermidis 0-47 grown in nutrient broth or in the presence of human serum. sdrG transcript levels increased 1 hour following a shift from growth in nutrient broth to growth in the bloodstream of a mouse and resulted in a concomitant increase in protein levels as detected by immunofluorescence microscopy. The environmental signal(s) resulting in the increase in expression is elusive, as growth under conditions known to mimic in vivo conditions (elevated CO(2), iron limitation, human serum, and citrated human blood) did not affect expression of SdrG. Immunizing mice with either the N1N2N3 (amino acids 50 to 597) or N2N3 (amino acids 273 to 597) subdomain of the N-terminal A domain of recombinant SdrG (rSdrG) elicited a robust antibody response; however, only mice vaccinated with rSdrG(N23) exhibited a significant reduction in 0-47 recovered after experimental infection. Since SdrG is expressed early during infection in response to specific host environmental cues present in the bloodstream and since antibodies to it are effective in reducing bacteremia, SdrG possesses attributes of a vaccine component effective against the pathogenic form of the ubiquitous human commensal S. epidermidis. PMID- 18426875 TI - Pilot-scale production and characterization of paramyosin, a vaccine candidate for schistosomiasis japonica. AB - Despite effective chemotherapy, schistosomiasis remains a major public health problem in the developing world, with at least 200 million active infections resulting in significant morbidity. Rapid reinfection after treatment, accompanied by extensive residual morbidity, mandates alternative control strategies, including vaccine development. Paramyosin, a myofibrillar protein found only in invertebrates, has been widely studied as a vaccine candidate for both Schistosoma mansoni and Schistosoma japonicum. Recently, we demonstrated that Th2-biased immune responses to paramyosin are associated with resistance to reinfection with S. japonicum in humans; however, challenges in the pilot-scale production of schistosome paramyosin have hampered further studies of this promising vaccine candidate. Here we report a method for the pilot-scale expression and purification of recombinant S. japonicum paramyosin (rSj97). rSj97 was extracted from Escherichia coli inclusion bodies and purified with sequential anion-exchange, hydroxyapatite, and size exclusion chromatography. The purified rSj97 was >95% pure as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoretic analysis and was free of significant endotoxin contamination. We demonstrate that, like native paramyosin, rSj97 adopts an alpha-helical coiled coil tertiary structure and binds immunoglobulin and collagen. Naive mice infected with S. japonicum produce anti-rSj97 immunoglobulin G (IgG) antibodies as early as 4 weeks postinfection, while sera collected from S. japonicum infected individuals contain anti-rSj97 IgE antibodies. Our method for pilot scale production of recombinant full-length paramyosin will facilitate preclinical evaluation of paramyosin as a vaccine for schistosomiasis. PMID- 18426876 TI - CCL28 is increased in human Helicobacter pylori-induced gastritis and mediates recruitment of gastric immunoglobulin A-secreting cells. AB - Human Helicobacter pylori infection gives rise to an active chronic gastritis and is a major risk factor for the development of duodenal ulcer disease and gastric adenocarcinoma. The infection is accompanied by a large accumulation of immunoglobulin A (IgA)-secreting cells in the gastric mucosa, and following mucosal immunization only H. pylori-infected volunteers mounted a B-cell response in the gastric mucosa. To identify the signals for recruitment of gastric IgA secreting cells, we investigated the gastric production of CCL28 (mucosa associated epithelial chemokine) and CCL25 (thymus-expressed chemokine) in H. pylori-infected and uninfected individuals and the potential of gastric B-cell populations to migrate toward these chemokines. Gastric tissue from H. pylori infected individuals contained significantly more CCL28 protein and mRNA than that from uninfected individuals, while CCL25 levels remained unchanged. Chemokine-induced migration of gastric lamina propria lymphocytes isolated from patients undergoing gastric resection was then assessed using the Transwell system. IgA-secreting cells and IgA(+) memory B cells from H. pylori-infected tissues migrated toward CCL28 but not CCL25, while the corresponding cells from uninfected patients did not. Furthermore, IgG-secreting cells from H. pylori infected patients did not migrate to CCL28 but instead to CXCL12 (SDF-1alpha). However, chemokine receptor expression did not correlate to the migratory pattern of the different B-cell populations. These studies are the first to show increased CCL28 production during gastrointestinal infection in humans and provide an explanation for the large influx of IgA-secreting cells to the gastric mucosa in H. pylori-infected individuals. PMID- 18426877 TI - Toll-like receptor 9 regulates the lung macrophage phenotype and host immunity in murine pneumonia caused by Legionella pneumophila. AB - Experiments were performed to determine the contribution of TLR9 to the generation of protective immunity against the intracellular respiratory bacterial pathogen Legionella pneumophila. In initial studies, we found that the intratracheal (i.t.) administration of L. pneumophila to mice deficient in TLR9 (TLR9(-/-)) resulted in significantly increased mortality, which was associated with an approximately 10-fold increase in the number of lung CFU compared to that of wild-type BALB/c mice. Intrapulmonary bacterial challenge in TLR9(-/-) mice resulted in the reduced accumulation of myeloid dendritic cells (DC) and activated CD4(+) T cells. Lung macrophages isolated from Legionella-infected TLR9(-/-) mice displayed the impaired internalization of bacteria and evidence of alternative rather than classical activation, as manifested by the markedly reduced expression of nitric oxide and type 1 cytokines, whereas the expression of Fizz-1 and arginase-1 was enhanced. The adoptive transfer of bone marrow derived DC from syngeneic wild-type, but not TLR9(-/-), mice administered i.t. reconstituted anti-legionella immunity and restored the macrophage phenotype in TLR9(-/-) mice. Finally, the i.t., but not intraperitoneal, administration of the TLR9 agonist molecule CpG oligodeoxynucleotide stimulated protective immunity in Legionella-infected mice. In total, our findings indicate that TLR9 is required for effective innate immune responses against the intracellular bacterial pathogen L. pneumophila, and approaches to maximize TLR9-mediated responses may serve as a means to augment antibacterial immunity in pneumonia. PMID- 18426878 TI - Immunogenicity and protective efficacy of recombinant Campylobacter jejuni flagellum-secreted proteins in mice. AB - Immunogenicity and protective efficacy of three Campylobacter jejuni flagellum secreted proteins, FlaC, FspA1, and FspA2, were compared by use of a mouse model. Mice were immunized intranasally with each protein with or without LTR192G as the adjuvant and challenged intranasally with C. jejuni 81-176 or CG8486. All three proteins were immunogenic, although FspA1 induced the highest levels of serum immunoglobulin G (IgG) and fecal IgA. Although immunogenic, FlaC provided only 18% protection against disease from C. jejuni 81-176. Immunization with FspA1 resulted in 57.8% protection without adjuvant or 63.8% protection with adjuvant against homologous challenge with 81-176. Alternatively, immunization with FspA2 provided 38.4% (without adjuvant) or 47.2% (with adjuvant) protection against disease from homologous challenge with CG8486. In contrast to FspA2, FspA1 provided some heterologous protection against C. jejuni CG8486 when delivered with (31.2%) or without (44.8%) LTR192G. These results suggest that FspA1 may be a good subunit vaccine candidate against C. jejuni disease. PMID- 18426879 TI - Significant role for ladC in initiation of Legionella pneumophila infection. AB - Previously, we identified ladC in a cohort of genes that were present in Legionella pneumophila but absent in other Legionella species. Here we constructed a ladC mutant of L. pneumophila and assessed its ability to replicate in mammalian cell lines and Acanthamoeba castellanii. The ladC mutant was recovered in significantly lower numbers than wild-type L. pneumophila at early time points, which was reversed upon transcomplementation with ladC but not ladC(N430A/R434A), encoding a putative catalytically inactive derivative of the protein. In fact, complementation of ladC::Km with ladC(N430A/R434A) resulted in a severe replication defect within human and amoeba cell models of infection, which did not follow a typical dominant negative phenotype. Using differential immunofluorescence staining to distinguish adherent from intracellular bacteria, we found that the ladC mutant exhibited a 10-fold reduction in adherence to THP-1 macrophages but no difference in uptake by THP-1 cells. When tested in vivo in A/J mice, the competitive index of the ladC mutant dropped fivefold over 72 h, indicating a significant attenuation compared to wild-type L. pneumophila. Although localization of LadC to the bacterial inner membrane suggested that the protein may be involved in signaling pathways that regulate virulence gene expression, microarray analysis indicated that ladC does not influence the transcriptional profile of L. pneumophila in vitro or during A. castellanii infection. Although the mechanism by which LadC modulates the initial interaction between the bacterium and host cell remains unclear, we have established that LadC plays an important role in L. pneumophila infection. PMID- 18426880 TI - Comparison of the contributions of heat-labile enterotoxin and heat-stable enterotoxin b to the virulence of enterotoxigenic Escherichia coli in F4ac receptor-positive young pigs. AB - In swine, the most common and severe enterotoxigenic Escherichia coli (ETEC) infections are caused by strains that express K88 (F4)(+) fimbriae, heat-labile enterotoxin (LT), heat-stable enterotoxin b (STb), and enteroaggregative E. coli heat-stable toxin 1. Previous studies based on a design that involved enterotoxin genes cloned into a nontoxigenic fimbriated strain have suggested that LT but not STb plays an important role in dehydrating diarrheal disease in piglets <1 week old and also enhances bacterial colonization of the intestine. In the present study, we compared these two toxins in terms of importance for piglets >1 week old with a study design that involved construction of isogenic single- and double deletion mutants and inoculation of 9-day-old F4ac receptor-positive gnotobiotic piglets. Based on the postinoculation percent weight change per h and serum bicarbonate concentrations, the virulence of the STb(-) mutant (Delta estB) did not significantly differ from that of the parent. However, deletion of the LT genes (Delta eltAB) in the STb(-) mutant resulted in a complete abrogation of weight loss, dehydration, and metabolic acidosis in inoculated pigs, and LT complementation restored the virulence of this strain. These results support the hypothesis that LT is a more significant contributor than STb to the virulence of F4(+) ETEC infections in young F4ac receptor-positive pigs less than 2 weeks old. However, in contrast to previous studies with gnotobiotic piglets, there was no evidence that the expression of LT enhanced the ability of the F4(+) ETEC strain to colonize the small intestine. PMID- 18426881 TI - Nasal immunization with Porphyromonas gingivalis outer membrane protein decreases P. gingivalis-induced atherosclerosis and inflammation in spontaneously hyperlipidemic mice. AB - Porphyromonas gingivalis has been shown to accelerate atherosclerotic lesion development in hyperlipidemic animals. We assessed the potential of a nasal vaccine against P. gingivalis infection for the prevention of atherosclerosis. Apolipoprotein E-deficient spontaneously hyperlipidemic (Apoe(shl)) mice were nasally immunized with the 40-kDa outer membrane protein (OMP) of P. gingivalis plus cholera toxin (CT) as adjuvant and then challenged intravenously with P. gingivalis strain 381. The animals were euthanized 11 or 14 weeks later. Atheromatous lesions in the proximal aorta of each animal were analyzed histomorphometrically, and the serum concentrations of 40-kDa OMP-specific antibodies and cytokines were determined. The areas of the aortic sinus that were covered with atherosclerotic plaque and the serum levels of inflammatory cytokines and chemokines were increased in Apoe(shl) mice challenged with P. gingivalis compared to nonchallenged mice. In comparison, nasal immunization with 40-kDa OMP plus CT significantly reduced atherosclerotic plaque accumulation in the aortic sinus and lowered the serum levels of cytokines and chemokines compared to nonimmunized animals. Nasal immunization also induced 40-kDa OMP specific serum immunoglobulin G (IgG) and saliva IgA antibody responses. These findings suggest that systemic infection with P. gingivalis accelerates atherosclerosis in Apoe(shl) mice, and 40-kDa OMP plus CT may be an effective nasal vaccine for the reduction of atherosclerosis accelerated by P. gingivalis in the hyperlipidemic mouse model. PMID- 18426882 TI - Role of aromatic amino acids in receptor binding activity and subunit assembly of the cytolethal distending toxin of Aggregatibacter actinomycetemcomitans. AB - The periodontal pathogen Aggregatibacter actinomycetemcomitans produces a cytolethal distending toxin (Cdt) that inhibits the proliferation of oral epithelial cells. Structural models suggest that the CdtA and CdtC subunits of the Cdt heterotrimer form two putative lectin domains with a central groove. A region of CdtA rich in heterocyclic amino acids (aromatic patch) appears to play an important role in receptor recognition. In this study site-specific mutagenesis was used to assess the contributions of aromatic amino acids (tyrosine and phenylalanine) to receptor binding and CdtA-CdtC assembly. Predominant surface-exposed aromatic residues that are adjacent to the aromatic patch region in CdtA or are near the groove located at the junction of CdtA and CdtC were studied. Separately replacing residues Y105, Y140, Y188, and Y189 with alanine in CdtA resulted in differential effects on binding related to residue position within the aromatic region. The data indicate that an extensive receptor binding domain extends from the groove across the entire face of CdtA that is oriented 180 degrees from the CdtB subunit. Replacement of residue Y105 in CdtA and residues Y61 and F141 in CdtC, which are located in or at the periphery of the groove, inhibited toxin assembly. Taken together, these results, along with the lack of an aromatic amino acid-rich region in CdtC similar to that in CdtA, suggest that binding of the heterotoxin to its cell surface receptor is mediated predominantly by the CdtA subunit. These findings are important for developing strategies designed to block the activity of this prominent virulence factor. PMID- 18426883 TI - Type IV pili in Francisella tularensis: roles of pilF and pilT in fiber assembly, host cell adherence, and virulence. AB - Francisella tularensis, a highly virulent facultative intracellular bacterium, is the causative agent of tularemia. Genome sequencing of all F. tularensis subspecies revealed the presence of genes that could encode type IV pili (Tfp). The live vaccine strain (LVS) expresses surface fibers resembling Tfp, but it was not established whether these fibers were indeed Tfp encoded by the pil genes. We show here that deletion of the pilF putative Tfp assembly ATPase in the LVS resulted in a complete loss of surface fibers. Disruption of the pilT putative disassembly ATPase also caused a complete loss of pili, indicating that pilT functions differently in F. tularensis than in model Tfp systems such as those found in Pseudomonas aeruginosa and Neisseria spp. The LVS pilF and pilT mutants were attenuated for virulence in a mouse model of tularemia by the intradermal route. Furthermore, although absence of pili had no effect on the ability of the LVS to replicate intracellularly, the pilF and pilT mutants were defective for adherence to macrophages, pneumocytes, and hepatocytes. This work confirms that the surface fibers expressed by the LVS are encoded by the pil genes and provides evidence that the Francisella pili contribute to host cell adhesion and virulence. PMID- 18426884 TI - The inositol phosphatase SHIP controls Salmonella enterica serovar Typhimurium infection in vivo. AB - The SH2 domain-containing inositol 5'-phosphatase, SHIP, negatively regulates various hematopoietic cell functions and is critical for maintaining immune homeostasis. However, whether SHIP plays a role in controlling bacterial infections in vivo remains unknown. Salmonella enterica causes human salmonellosis, a disease that ranges in severity from mild gastroenteritis to severe systemic illness, resulting in significant morbidity and mortality worldwide. The susceptibility of ship(+/+) and ship(-/-) mice and bone marrow derived macrophages to S. enterica serovar Typhimurium infection was compared. ship(-/-) mice displayed an increased susceptibility to both oral and intraperitoneal serovar Typhimurium infection and had significantly higher bacterial loads in intestinal and systemic sites than ship(+/+) mice, indicating a role for SHIP in the gut-associated and systemic pathogenesis of serovar Typhimurium in vivo. Cytokine analysis of serum from orally infected mice showed that ship(-/-) mice produce lower levels of Th1 cytokines than do ship(+/+) animals at 2 days postinfection, and in vitro analysis of supernatants taken from infected bone marrow-derived macrophages derived to mimic the in vivo ship(-/-) alternatively activated (M2) macrophage phenotype correlated with these data. M2 macrophages were the predominant population in vivo in both oral and intraperitoneal infections, since tissue macrophages within the small intestine and peritoneal macrophages from ship(-/-) mice showed elevated levels of the M2 macrophage markers Ym1 and Arginase 1 compared to ship(+/+) cells. Based on these data, we propose that M2 macrophage skewing in ship(-/-) mice contributes to ineffective clearance of Salmonella in vivo. PMID- 18426885 TI - The cytosolic pattern recognition receptor NOD1 induces inflammatory interleukin 8 during Chlamydia trachomatis infection. AB - Inflammation is a hallmark of chlamydial infections, but how inflammatory cytokines are induced is not well understood. Pattern recognition receptors (PRR) of the host innate immune system recognize pathogen molecules and activate intracellular signaling pathways that modulate immune responses. The role of PRR such as Toll-like receptors (TLR) and nucleotide-binding oligomerization domain (NOD) proteins in the endogenous interleukin-8 (IL-8) response induced during Chlamydia trachomatis infection is not known. We hypothesized that a PRR is essential for the IL-8 response induced by C. trachomatis infection. RNA interference was used to knock down the TLR signaling partner MyD88 as well as NOD1 and its signaling molecule receptor-interacting protein 2 (RIP2). IL-8 induced at 30 h postinfection by C. trachomatis was dependent on NOD1 signaling through RIP2; however, the IL-8 response was independent of MyD88-dependent TLR signaling. Activation of the extracellular signal-regulated kinase (ERK) mitogen activated protein kinase cellular signaling pathway, which is essential for up regulation of IL-8 in response to C. trachomatis infection, was independent of NOD1 or RIP2. We conclude that the endogenous IL-8 response induced by C. trachomatis infection is dependent upon NOD1 PRR signaling through RIP2 as part of a signal system requiring multiple inputs for optimal IL-8 induction. Since ERK is not activated through this pathway, a concomitant interaction between the host and bacteria is additionally required for full activation of the endogenous IL-8 response. PMID- 18426886 TI - Suppression of T-lymphocyte activation and chemotaxis by the adenylate cyclase toxin of Bordetella pertussis. AB - The adenylate cyclase toxin (CyaA) released by Bordetella pertussis is an essential virulence factor for colonization of the host. This toxin inhibits migration and activation of phagocytes, thereby preventing bacterial killing. In addition, CyaA interferes with the initiation of adaptive immunity by misdirecting dendritic cell differentiation to a suppressive rather than stimulatory phenotype. Here we show that CyaA directly affects adaptive responses by catalyzing cyclic AMP (cAMP) production in peripheral blood lymphocytes. Treatment with CyaA resulted in profound impairment of T-lymphocyte activation and chemotaxis. These effects resulted from inhibition of T-cell antigen receptor and chemokine receptor signaling via a cAMP/protein kinase A (PKA)-dependent pathway. A comparison of the activities of CyaA on T-cell and macrophage activation and migration revealed that the biological effects of the toxin were paralleled by inhibition of the activation of mitogen-activated protein (MAP) kinases, highlighting the conclusion that the ubiquitous and evolutionarily conserved MAP kinase modules are common targets of the PKA-mediated immunosuppressant activities of CyaA and underlining the potential of cAMP elevating toxins as a means of evasion of immunity by bacterial pathogens. PMID- 18426887 TI - Metal binding specificity of the MntABC permease of Neisseria gonorrhoeae and its influence on bacterial growth and interaction with cervical epithelial cells. AB - mntABC from Neisseria gonorrhoeae encodes an ABC permease which includes a periplasmic divalent cation binding receptor protein of the cluster IX family, encoded by mntC. Analysis of an mntC mutant showed that growth of N. gonorrhoeae could be stimulated by addition of either manganese(II) or zinc(II) ions, suggesting that the MntABC system could transport both ions. In contrast, growth of the mntAB mutant in liquid culture was possible only when the medium was supplemented with an antioxidant such as mannitol, consistent with the view that ion transport via MntABC is essential for protection of N. gonorrhoeae against oxidative stress. Using recombinant MntC, we determined that MntC binds Zn(2+) and Mn(2+) with almost equal affinity (dissociation constant of approximately 0.1 microM). Competition assays with the metallochromic zinc indicator 4-(2 pyridylazo)resorcinol showed that MntC binds Mn(2+) and Zn(2+) at the same binding site. Analysis of the N. gonorrhoeae genome showed that MntC is the only Mn/Zn metal binding receptor protein cluster IX in this bacterium, in contrast to the situation in many other bacteria which have systems with dedicated Mn and Zn binding proteins as part of distinctive ABC cassette permeases. Both the mntC and mntAB mutants had reduced intracellular survival in a human cervical epithelial cell model and showed reduced ability to form a biofilm. These data suggest that the MntABC transporter is of importance for survival of Neisseria gonorrhoeae in the human host. PMID- 18426888 TI - Incorporation of Mycobacterium tuberculosis lipoarabinomannan into macrophage membrane rafts is a prerequisite for the phagosomal maturation block. AB - Lipoarabinomannan (LAM) is one of the key virulence factors for Mycobacterium tuberculosis, the etiological agent of tuberculosis. During uptake of mycobacteria, LAM interacts with the cell membrane of the host macrophage and can be detected throughout the cell upon infection. LAM can inhibit phagosomal maturation as well as induce a proinflammatory response in bystander cells. The aim of this study was to investigate how LAM exerts its action on human macrophages. We show that LAM is incorporated into membrane rafts of the macrophage cell membrane via its glycosylphosphatidylinositol anchor and that incorporation of mannose-capped LAM from M. tuberculosis results in reduced phagosomal maturation. This is dependent on successful insertion of the glycosylphosphatidylinositol anchor. LAM does not, however, induce the phagosomal maturation block through activation of p38 mitogen-activated protein kinase, contradicting some previous suggestions. PMID- 18426889 TI - Impact of mating type, serotype, and ploidy on the virulence of Cryptococcus neoformans. AB - Hybridization with polyploidization is a significant biological force driving evolution. The effect of combining two distinct genomes in one organism on the virulence potential of pathogenic fungi is not clear. Cryptococcus neoformans, the most common cause of fungal infection of the central nervous system, has a bipolar mating system with a and alpha mating types and occurs as A (haploid), D (haploid), and AD hybrid (mostly diploid) serotypes. Diploid AD hybrids are derived either from a-alpha mating or from unisexual mating between haploid cells. The precise contributions of increased ploidy, the effect of hybridization between serotypes A and D, and the combination of mating types to the virulence potential of AD hybrids have remained elusive. By using in vitro and in vivo characterization of laboratory-constructed isogenic diploids and AD hybrids with all possible mating type combinations in defined genetic backgrounds, we found that higher ploidy has a minor negative effect on virulence in a murine inhalation model of cryptococcosis. The presence of both mating types a and alpha in AD hybrids did not affect the virulence potential, irrespective of the serotype origin. Interestingly, AD hybrids with only one mating type behaved differently, with the virulence of alphaADalpha strains similar to that of other hybrids, while aADa hybrids displayed significantly lower virulence due to negative epistatic interactions between the Aa and Da alleles of the mating type locus. This study provides insights into the impact of ploidy, mating type, and serotype on virulence and the impact of hybridization on the fitness and virulence of a eukaryotic microbial pathogen. PMID- 18426890 TI - The tick salivary protein Salp15 inhibits the killing of serum-sensitive Borrelia burgdorferi sensu lato isolates. AB - Borrelia burgdorferi, the agent of Lyme disease, is transmitted by ticks. During transmission from the tick to the host, spirochetes are delivered with tick saliva, which contains the salivary protein Salp15. Salp15 has been shown to protect spirochetes against B. burgdorferi-specific antibodies. We now show that Salp15 from both Ixodes ricinus and Ixodes scapularis protects serum-sensitive isolates of Borrelia burgdorferi sensu lato against complement-mediated killing. I. ricinus Salp15 showed strong protective effects compared to those of I. scapularis Salp15. Deposition of terminal C5b to C9 (one molecule each of C5b, C6, C7, and C8 and one or more molecules of C9) complement complexes, part of the membrane attack complex, on the surface of B. burgdorferi was inhibited in the presence of Salp15. In the presence of normal human serum, serum-sensitive Borrelia burgdorferi requires protection against complement-mediated killing, which is provided, at least in part, by the binding to the tick salivary protein Salp15. PMID- 18426891 TI - tRNA modification by GidA/MnmE is necessary for Streptococcus pyogenes virulence: a new strategy to make live attenuated strains. AB - Studies directed at vaccine development and mucosal immunity against Streptococcus pyogenes would benefit from the availability of live attenuated strains. Our approach for production of candidate live attenuated strains was to identify mutations that did not alter growth in vitro and did not alter the overall complement of virulence factors produced but did result in reduced levels of expression of multiple secreted virulence factors. A global reduction but not elimination of expression would likely lead to attenuation while maximizing the number of antigenic targets available for stimulation of immunity. Adaptation of Tn5-based transposome mutagenesis to S. pyogenes with initial screening for reduced expression of the SpeB protease resulted in identification of mutations in gidA, which encodes an enzyme involved in tRNA modification. Reduced SpeB expression was due to delayed onset of speB transcription resulting from reduced translation efficiency of the message for RopB, a transcriptional activator. Overall, GidA(-) mutants had a nearly normal global transcription profile but expressed significantly reduced levels of multiple virulence factors due to impaired translation efficiencies. A translation defect was supported by the observation that mutants lacking MnmE, which functions in the same tRNA modification pathway as GidA, phenocopied GidA deficiency. The mutants stimulated a cytokine response in cultured macrophages identical to that in the wild type, with the exception of reduced levels of tumor necrosis factor alpha and interleukin-23. Significantly, GidA(-) mutants were highly attenuated in the murine ulcer model of soft tissue infection. These characteristics suggest that GidA pathway tRNA modification mutants are attractive candidates for further evaluation as live attenuated strains. PMID- 18426892 TI - Novel role of sphingolipid synthesis genes in regulating giardial encystation. AB - Although encystation (cyst formation) is important for the survival of Giardia lamblia outside its human host, the molecular events that prompt encystation have not been fully elucidated. Here, we demonstrate that sphingolipids (SLs), which are important for the growth and differentiation of many eukaryotes, play key roles in giardial encystation. Transcriptional analyses showed that only three genes in the SL biosynthesis pathways are expressed and transcribed differentially in nonencysting and encysting Giardia trophozoites. While the putative homologues of giardial serine palmitoyltransferase (gSPT) subunit genes (gspt-1 and -2) are differentially expressed in nonencysting and encysting trophozoites, the giardial ceramide glucosyltransferase 1 gene (gglct-1) is transcribed only in encysting cells. l-Cycloserine, an inhibitor of gSPT, inhibited the endocytosis and endoplasmic reticulum/perinuclear targeting of bodipy-ceramide in trophozoites, and this could be reversed by 3-ketosphinganine. On the other hand, D-threo-1-phenyl-2-palmitoylamino-3-morpholino-1-propanol (PPMP), an inhibitor of glucosylceramide synthesis, blocked karyokinesis and reduced cyst production in culture. PPMP also altered the expression of cyst wall protein transcripts in encysting cells. Phylogenetic analyses revealed that the gspt genes are paralogs derived from an ancestral spt sequence that underwent gene duplication early in eukaryotic history. This ancestral sequence, in turn, was probably derived from prokaryotic aminoacyl transferases. In contrast, gglct 1 is found in both prokaryotes and eukaryotes without any evidence of gene duplication. These studies indicate that SL synthesis genes are involved in key events in giardial biology and could serve as potential targets for developing new therapies against giardiasis. PMID- 18426893 TI - Systemic expression of cytokine production in patients with severe pneumococcal pneumonia: effects of treatment with a beta-lactam versus a fluoroquinolone. AB - Bacterial alveolar invasion is followed by an inflammatory response. A systemic extension of the compartmentalized immune response has been described in patients with severe pneumonia. The data suggest that some antimicrobials may induce a differential release of cytokines. We conducted a prospective, randomized study in adult patients with severe pneumococcal pneumonia to measure the effects of ceftriaxone and levofloxacin in the systemic cytokine expression over time. Demographic, clinical characteristics, and severity scores were recorded. The serum concentrations of tumor necrosis factor alpha (TNF-alpha), interleukin 1beta (IL-1beta), IL-6, IL-8, IL-10, and IL-1 receptor agonist were measured at 0, 24, 72, and 120 h. A total of 32 patients were included in the study. Both groups were homogeneous in terms of age, comorbidities, severity of disease, and corticosteroid or statin use. With the single exception of IL-1beta, all cytokines were detected in venous blood. All of the cytokines studied showed a similar pattern of progressive decrease over time. No significant differences in the concentrations of any of the cytokines studied were found, with the exception of TNF-alpha, for which lower concentrations were obtained at 120 h in the levofloxacin group (P = 0.014). Basal oxygen saturation (P = 0.034) and heart rate (P = 0.029) returned to normal values earlier in the levofloxacin arm. We demonstrated that in patients with severe pneumococcal pneumonia pro- and anti inflammatory responses could be detected in venous blood, representing a systemic extension of the compartmentalized response. Treatment with a beta-lactam agent or a fluoroquinolone has different effects on cytokine production and its systemic expression, impacting the clinical course of the disease. PMID- 18426894 TI - qnr Gene nomenclature. PMID- 18426895 TI - Impact of mycophenolic acid on the functionality of human polymorphonuclear neutrophils and dendritic cells during interaction with Aspergillus fumigatus. AB - Mycophenolic acid (MPA) is used clinically to prevent graft rejection but may increase the risk of fungal infection. We observed that MPA enhanced the Aspergillus fumigatus-induced oxidative burst of polymorphonuclear neutrophils, but without a corresponding increase in fungal killing. Furthermore, MPA inhibited the proinflammatory cytokine response and maturation of dendritic cells. PMID- 18426896 TI - Activities and ultrastructural effects of antifungal combinations against simulated Candida endocardial vegetations. AB - In vitro pharmacodynamic model (PDM) simulation of serum antifungal concentrations may predict the value of combination antifungal regimens against Candida sp. endocarditis. We investigated the effects of combinations of flucytosine (5FC), micafungin (Mica), and voriconazole (Vor) against Candida infected human platelet-fibrin clots, used as simulated endocardial vegetations (SEVs). Single clinical bloodstream isolates of Candida albicans, Candida glabrata, Candida parapsilosis, and Candida tropicalis were used. All four isolates were susceptible to 5FC, while C. glabrata was resistant to Vor and C. tropicalis had a paradoxical resistance phenotype to Mica. The SEVs were prepared with an initial inoculum of 1 x 10(6) CFU/g of SEV and added to a PDM, which utilized yeast nitrogen broth-2% glucose and incubation at 35 degrees C and simulated antifungal pharmacokinetic profiles. Fungal densities in the SEVs were determined in quadruplicate over 72 h. Scanning electron microscopy (SEM) was used to evaluate treatment and control SEVs. Vor was the least active single agent against all Candida spp. except for C. parapsilosis, where it was comparable to Mica. In contrast, 5FC was the most active against all Candida spp. except for C. tropicalis, where it was comparable to Mica. The combination of 5FC plus Vor was superior to either agent alone against C. parapsilosis. The combination of Vor plus Mica was inferior to the use of Mica alone against C. tropicalis. The triple combination of 5FC plus Vor plus Mica was no better than single or dual agents against any of the Candida spp. The ultrastructural features of infected SEVs were unique for each Candida sp., with C. parapsilosis in particular manifesting friable biofilm clusters. In general, 5FC and Mica were superior in their rates and extents of fungal burden reduction compared to Vor against Candida-infected SEVs. Evaluation of 5FC and Mica in animal models of Candida endocarditis is warranted. PMID- 18426897 TI - High levels of zidovudine (AZT) and its intracellular phosphate metabolites in AZT- and AZT-lamivudine-treated newborns of human immunodeficiency virus-infected mothers. AB - Newborns from human immunodeficiency virus-infected mothers are given antiretroviral prophylaxis against mother-to-child transmission, including predominantly nucleoside reverse transcriptase inhibitors. Pharmacological monitoring of these drugs in newborns has so far been limited to plasma and cord blood. In this study, samples from newborns (up to 45 days old) treated with zidovudine (AZT) alone (n = 29) or in combination with lamivudine (3TC) (n = 20) were analyzed for both intracellular concentrations of phosphate metabolites in peripheral blood mononuclear cells and levels of parent drugs in plasma. Plasma AZT and intracellular AZT-monophosphate and AZT-triphosphate (TP) concentrations were significantly higher during the first 15 days of life (199 versus 52.7 ng/ml [P < 0.0001], 732 versus 282 fmol/10(6) cells [P < 0.0001], and 170 versus 65.1 fmol/10(6) cells [P < 0.0001], respectively) and then became comparable to those of adults. No difference in intracellular AZT metabolite concentrations was found when AZT- and AZT-3TC-treated groups were compared. Plasma 3TC levels (lower limit of quantification [LLOQ], 1,157 ng/ml; median, 412.5 ng/ml) were not associated with the newborn's age, gender, or weight. Intracellular 3TC-TP concentrations (LLOQ, 40.4 pmol/10(6) cells; median, 18.9 pmol/10(6) cells) determined for newborns receiving the AZT-3TC combination were associated with neither the age nor weight of the newborns. Concentrations in females were significantly higher (1.8-fold [P = 0.0415]) than those in males. Unexpectedly, newborns on AZT monotherapy whose mothers' treatment included 3TC displayed residual plasma 3TC and intracellular 3TC-TP levels up to 1 week after birth. PMID- 18426898 TI - Telavancin penetration into human epithelial lining fluid determined by population pharmacokinetic modeling and Monte Carlo simulation. AB - Telavancin is an investigational bactericidal lipoglycopeptide with a multifunctional mechanism of action, as demonstrated against methicillin resistant Staphylococcus aureus. While the plasma pharmacokinetics have been described, the extent of the penetration of the drug into the lung, measured by the epithelial lining fluid (ELF), remains unknown. Population modeling and Monte Carlo simulation were employed to estimate the penetration of telavancin into ELF. Plasma and ELF pharmacokinetic data were obtained from 20 healthy volunteers, and the pharmacokinetic samples were assayed by a validated liquid chromatography-tandem mass spectrometry technique. Concentration-time profiles in plasma and ELF were simultaneously modeled using a three-compartment model with zero-order infusion and first-order elimination and transfer. The model parameters were identified in a population pharmacokinetic analysis (BigNPAG). Monte Carlo simulation of 9,999 subjects was performed to calculate the ELF/plasma penetration ratios by estimating the area under the concentration-time curve (AUC) for the drug in ELF (AUC(ELF)) and for the free drug in plasma (free AUC(plasma)) from zero to infinity after a single dose. After the Bayesian step, the overall fits of the model to the data were good, and plots of predicted versus observed concentrations in plasma and ELF showed slopes and intercepts very close to the ideal values of 1.0 and 0.0, respectively. The median AUC(ELF)/free AUC(plasma) penetration ratio was 0.73, and the 25th and 75th percentile value ratios were 0.43 and 1.24, respectively. In uninfected lung tissue, the median AUC(ELF) is approximately 75% of the free AUC(plasma). PMID- 18426899 TI - Presence of plasmid-mediated quinolone resistance in Klebsiella pneumoniae isolates possessing blaKPC in the United States. AB - The presence of plasmid-mediated quinolone resistance genes [i.e., qnrA, qnrB, qnrS, aac(6')-Ib-cr, and qepA] was evaluated among 42 bla(KPC)-containing Klebsiella pneumoniae isolates collected in the eastern United States. One isolate carried the bla(KPC-3) and qnrB19 genes on the same conjugative plasmid, whereas another carried the bla(KPC-3) and qnrA1 genes on separate plasmids. PMID- 18426900 TI - Daptomycin is effective in treatment of experimental endocarditis due to methicillin-resistant and glycopeptide-intermediate Staphylococcus aureus. AB - Daptomycin is a lipopeptide antibiotic with potent in vitro activity against gram positive cocci, including Staphylococcus aureus. This study evaluated the in vitro and in vivo efficacies of daptomycin against two clinical isolates: methicillin-resistant S. aureus (MRSA) 277 (vancomycin MIC, 2 microg/ml) and glycopeptide-intermediate S. aureus (GISA) ATCC 700788 (vancomycin MIC, 8 microg/ml). Time-kill experiments demonstrated that daptomycin was bactericidal in vitro against these two strains. The in vivo activity of daptomycin (6 mg/kg of body weight every 24 h) was evaluated by using a rabbit model of infective endocarditis and was compared with the activities of a high-dose (HD) vancomycin regimen (1 g intravenously every 6 h), the recommended dose (RD) of vancomycin regimen (1 g intravenously every 12 h) for 48 h, and no treatment (as a control). Daptomycin was significantly more effective than the vancomycin RD in reducing the density of bacteria in the vegetations for the MRSA strains (0 [interquartile range, 0 to 1.5] versus 2 [interquartile range, 0 to 5.6] log CFU/g vegetation; P = 0.02) and GISA strains (2 [interquartile range, 0 to 2] versus 6.6 [interquartile range, 2.0 to 6.9] log CFU/g vegetation; P < 0.01) studied. In addition, daptomycin sterilized more MRSA vegetations than the vancomycin RD (13/18 [72%] versus 7/20 [35%]; P = 0.02) and sterilized more GISA vegetations than either vancomycin regimen (12/19 [63%] versus 4/20 [20%]; P < 0.01). No statistically significant difference between the vancomycin HD and the vancomycin RD for MRSA treatment was noted. These results support the use of daptomycin for the treatment of aortic valve endocarditis caused by GISA and MRSA. PMID- 18426901 TI - Mutagenesis in the alpha3alpha4 GyrA helix and in the Toprim domain of GyrB refines the contribution of Mycobacterium tuberculosis DNA gyrase to intrinsic resistance to quinolones. AB - The replacement of M74 in GyrA, A83 in GyrA, and R447 in GyrB of Mycobacterium tuberculosis gyrase by their Escherichia coli homologs resulted in active enzymes as quinolone susceptible as the E. coli gyrase. This demonstrates that the primary structure of gyrase determines intrinsic quinolone resistance and was supported by a three-dimensional model of N-terminal GyrA. PMID- 18426902 TI - In vitro antibacterial activity of CE-156811, a novel analog derived from hygromycin A. AB - We evaluated a novel truncated hygromycin A analog in which the furanose ring was replaced with a 2-fluoro-2-cyclopropylethyl substituent for its activity against multidrug resistant gram-positive bacteria and compared its activity to the activities of linezolid, quinupristin-dalfopristin, and vancomycin. CE-156811 demonstrated robust in vitro activity against gram-positive bacteria that was comparable to that of linezolid. PMID- 18426903 TI - Identification of ancient remains through genomic sequencing. AB - Studies of ancient DNA have been hindered by the preciousness of remains, the small quantities of undamaged DNA accessible, and the limitations associated with conventional PCR amplification. In these studies, we developed and applied a genomewide adapter-mediated emulsion PCR amplification protocol for ancient mammalian samples estimated to be between 45,000 and 69,000 yr old. Using 454 Life Sciences (Roche) and Illumina sequencing (formerly Solexa sequencing) technologies, we examined over 100 megabases of DNA from amplified extracts, revealing unbiased sequence coverage with substantial amounts of nonredundant nuclear sequences from the sample sources and negligible levels of human contamination. We consistently recorded over 500-fold increases, such that nanogram quantities of starting material could be amplified to microgram quantities. Application of our protocol to a 50,000-yr-old uncharacterized bone sample that was unsuccessful in mitochondrial PCR provided sufficient nuclear sequences for comparison with extant mammals and subsequent phylogenetic classification of the remains. The combined use of emulsion PCR amplification and high-throughput sequencing allows for the generation of large quantities of DNA sequence data from ancient remains. Using such techniques, even small amounts of ancient remains with low levels of endogenous DNA preservation may yield substantial quantities of nuclear DNA, enabling novel applications of ancient DNA genomics to the investigation of extinct phyla. PMID- 18426904 TI - Comparative proteogenomics: combining mass spectrometry and comparative genomics to analyze multiple genomes. AB - Recent proliferation of low-cost DNA sequencing techniques will soon lead to an explosive growth in the number of sequenced genomes and will turn manual annotations into a luxury. Mass spectrometry recently emerged as a valuable technique for proteogenomic annotations that improves on the state-of-the-art in predicting genes and other features. However, previous proteogenomic approaches were limited to a single genome and did not take advantage of analyzing mass spectrometry data from multiple genomes at once. We show that such a comparative proteogenomics approach (like comparative genomics) allows one to address the problems that remained beyond the reach of the traditional "single proteome" approach in mass spectrometry. In particular, we show how comparative proteogenomics addresses the notoriously difficult problem of "one-hit-wonders" in proteomics, improves on the existing gene prediction tools in genomics, and allows identification of rare post-translational modifications. We therefore argue that complementing DNA sequencing projects by comparative proteogenomics projects can be a viable approach to improve both genomic and proteomic annotations. PMID- 18426905 TI - Downregulation of vertebrate Tel (ETV6) and Drosophila Yan is facilitated by an evolutionarily conserved mechanism of F-box-mediated ubiquitination. AB - The vertebrate Ets transcriptional repressor Tel (ETV6) and its invertebrate orthologue, Yan, are both indispensable for development, and they orchestrate cell growth and differentiation by binding to DNA, thus inhibiting gene expression. To trigger cell differentiation, these barriers to transcriptional activation must be relieved, and it is established that posttranslational modifications, such as phosphorylation and sumoylation, can specifically impair the repressive functions of Tel and Yan and are crucial for modulating their transcriptional activity. To date, however, relatively little is known about the control of Tel and Yan protein degradation. In recent years, there has been a concentrated effort to assign functions to the large number of F-box proteins encoded by both vertebrate and invertebrate genomes. Here, we report the identification and characterization of a previously unreported, evolutionarily conserved F-box protein named Fbl6. We isolated both human and Drosophila melanogaster fbl6 cDNA and show that the encoded Fbl6 protein binds to both Tel and Yan via their SAM domains. We demonstrate that both Tel and Yan are ubiquitinated, a process which is stimulated by Fbl6 and leads to proteasomal degradation. We recently established that the sumoylation of Tel on lysine 11 negatively regulates its repressive function and that the sumoylation of Tel monomers, but not that of Tel oligomers, may sensitize Tel for proteasomal degradation. Here, we found that Fbl6 regulates Tel/Yan protein stability and allows appropriate spatiotemporal control of gene expression by these repressors. PMID- 18426906 TI - Residues of Tim44 involved in both association with the translocon of the inner mitochondrial membrane and regulation of mitochondrial Hsp70 tethering. AB - Translocation of proteins from the cytosol across the mitochondrial inner membrane is driven by the action of the import motor, which is associated with the translocon on the matrix side of the membrane. It is well established that an essential peripheral membrane protein, Tim44, tethers mitochondrial Hsp70 (mtHsp70), the core of the import motor, to the translocon. This Tim44-mtHsp70 interaction, which can be recapitulated in vitro, is destabilized by binding of mtHsp70 to a substrate polypeptide. Here we report that the N-terminal 167-amino acid segment of mature Tim44 is sufficient for both interaction with mtHsp70 and destabilization of a Tim44-mtHsp70 complex caused by client protein binding. Amino acid alterations within a 30-amino-acid segment affected both the release of mtHsp70 upon peptide binding and the interaction of Tim44 with the translocon. Our results support the idea that Tim44 plays multiple roles in mitochondrial protein import by recruiting Ssc1 and its J protein cochaperone to the translocon and coordinating their interactions to promote efficient protein translocation in vivo. PMID- 18426907 TI - Aberrant expression of nucleostemin activates p53 and induces cell cycle arrest via inhibition of MDM2. AB - The nucleolar protein nucleostemin (NS) is essential for cell proliferation and early embryogenesis. Both depletion and overexpression of NS reduce cell proliferation. However, the mechanisms underlying this regulation are still unclear. Here, we show that NS regulates p53 activity through the inhibition of MDM2. NS binds to the central acidic domain of MDM2 and inhibits MDM2-mediated p53 ubiquitylation and degradation. Consequently, ectopic overexpression of NS activates p53, induces G(1) cell cycle arrest, and inhibits cell proliferation. Interestingly, the knockdown of NS by small interfering RNA also activates p53 and induces G(1) arrest. These effects require the ribosomal proteins L5 and L11, since the depletion of NS enhanced their interactions with MDM2 and the knockdown of L5 or L11 abrogated the NS depletion-induced p53 activation and cell cycle arrest. These results suggest that a p53-dependent cell cycle checkpoint monitors changes of cellular NS levels via the impediment of MDM2 function. PMID- 18426908 TI - Targeting the GA binding protein beta1L isoform does not perturb lymphocyte development and function. AB - GA binding protein (GABP) is a ubiquitously expressed Ets family transcription factor that consists of two subunits, GABPalpha and GABPbeta. GABPalpha binds to DNA, and GABPbeta heterodimerizes with GABPalpha and possesses the ability to transactivate target genes. Our previous studies using GABPalpha-deficient mice revealed that GABPalpha is required for the development of both T and B cells. Two splice variants of GABPbeta are generated from the Gabpb1 locus and differ in their carboxy-terminal lengths and sequences. The longer isoform (GABPbeta1L) can homodimerize and thus form alpha(2)beta(2) tetramers depending on the gene context, whereas the shorter isoform (GABPbeta1S) cannot. In this study, we generated mice that are deficient in GABPbeta1L but that retain the expression of GABPbeta1S. Surprisingly, GABPbeta1L-/- mice had normal T- and B-cell development, and mature T and B cells showed normal responses to various stimuli. In contrast, targeting both GABPbeta1L and GABPbeta1S resulted in early embryonic lethality. Because of its incapability of forming homodimers, GABPbeta1S has been suspected to have a dominant negative role in regulating GABP target genes. Our findings argue against such a possibility and rather suggest that GABPbeta1S has a critical role in maintaining the transcriptional activity of the GABPalpha/beta complex. PMID- 18426909 TI - Properties of an intergenic terminator and start site switch that regulate IMD2 transcription in yeast. AB - The IMD2 gene in Saccharomyces cerevisiae is regulated by intracellular guanine nucleotides. Regulation is exerted through the choice of alternative transcription start sites that results in synthesis of either an unstable short transcript terminating upstream of the start codon or a full-length productive IMD2 mRNA. Start site selection is dictated by the intracellular guanine nucleotide levels. Here we have mapped the polyadenylation sites of the upstream, unstable short transcripts that form a heterogeneous family of RNAs of approximately 200 nucleotides. The switch from the upstream to downstream start sites required the Rpb9 subunit of RNA polymerase II. The enzyme's ability to locate the downstream initiation site decreased exponentially as the start was moved downstream from the TATA box. This suggests that RNA polymerase II's pincer grip is important as it slides on DNA in search of a start site. Exosome degradation of the upstream transcripts was highly dependent upon the distance between the terminator and promoter. Similarly, termination was dependent upon the Sen1 helicase when close to the promoter. These findings extend the emerging concept that distinct modes of termination by RNA polymerase II exist and that the distance of the terminator from the promoter, as well as its sequence, is important for the pathway chosen. PMID- 18426910 TI - Caspase-2 cleavage of BID is a critical apoptotic signal downstream of endoplasmic reticulum stress. AB - The accumulation of misfolded proteins stresses the endoplasmic reticulum (ER) and triggers cell death through activation of the multidomain proapoptotic BCL-2 proteins BAX and BAK at the outer mitochondrial membrane. The signaling events that connect ER stress with the mitochondrial apoptotic machinery remain unclear, despite evidence that deregulation of this pathway contributes to cell loss in many human degenerative diseases. In order to "trap" and identify the apoptotic signals upstream of mitochondrial permeabilization, we challenged Bax-/- Bak-/- mouse embryonic fibroblasts with pharmacological inducers of ER stress. We found that ER stress induces proteolytic activation of the BH3-only protein BID as a critical apoptotic switch. Moreover, we identified caspase-2 as the premitochondrial protease that cleaves BID in response to ER stress and showed that resistance to ER stress-induced apoptosis can be conferred by inhibiting caspase-2 activity. Our work defines a novel signaling pathway that couples the ER and mitochondria and establishes a principal apoptotic effector downstream of ER stress. PMID- 18426911 TI - A Rictor-Myo1c complex participates in dynamic cortical actin events in 3T3-L1 adipocytes. AB - Insulin signaling through phosphatidylinositol 3-kinase (PI 3-kinase) activates the protein kinase Akt through phosphorylation of its threonine 308 and serine 473 residues by the PDK1 protein kinase and the Rictor-mammalian target of rapamycin complex (mTORC2), respectively. Remarkably, we show here that the Rictor protein is also present in cultured adipocytes in complexes containing Myo1c, a molecular motor that promotes cortical actin remodeling. Interestingly, the Rictor-Myo1c complex is biochemically distinct from the previously reported mTORC2 and can be immunoprecipitated independently of mTORC2. Furthermore, while RNA interference-directed silencing of Rictor results in the expected attenuation of Akt phosphorylation at serine 473, depletion of Myo1c is without effect. In contrast, loss of either Rictor or Myo1c inhibits phosphorylation of the actin filament regulatory protein paxillin at tyrosine 118. Furthermore, Myo1c-induced membrane ruffling of 3T3-L1 adipocytes is also compromised following Rictor knockdown. Interestingly, neither the mTORC2 inhibitor rapamycin nor the PI 3 kinase inhibitor wortmannin affects paxillin tyrosine 118 phosphorylation. Taken together, our findings suggest that the Rictor-Myo1c complex is distinct from mTORC2 and that Myo1c, in conjunction with Rictor, participates in cortical actin remodeling events. PMID- 18426912 TI - Hepatocyte nuclear factor 4alpha contributes to thyroid hormone homeostasis by cooperatively regulating the type 1 iodothyronine deiodinase gene with GATA4 and Kruppel-like transcription factor 9. AB - Type 1 iodothyronine deiodinase (Dio1), a selenoenzyme catalyzing the bioactivation of thyroid hormone, is highly expressed in the liver. Dio1 mRNA and enzyme activity levels are markedly reduced in the livers of hepatocyte nuclear factor 4alpha (HNF4alpha)-null mice, thus accounting for its liver-specific expression. Consistent with this deficiency, serum T4 and rT3 concentrations are elevated in these mice compared with those in HNF4alpha-floxed control littermates; however, serum T3 levels are unchanged. Promoter analysis of the mouse Dio1 gene demonstrated that HNF4alpha plays a key role in the transactivation of the mouse Dio1 gene. Deletion and substitution mutation analyses demonstrated that a proximal HNF4alpha site (direct repeat 1 [TGGACAAAGGTGC]; HNF4alpha-RE) is crucial for transactivation of the mouse Dio1 gene by HNF4alpha. Mouse Dio1 is also stimulated by thyroid hormone signaling, but a direct role for thyroid hormone receptor action has not been reported. We also showed that thyroid hormone-inducible Kruppel-like factor 9 (KLF9) stimulates the mouse Dio1 promoter very efficiently through two CACCC sequences that are located on either side of HNF4alpha-RE. Furthermore, KLF9 functions together with HNF4alpha and GATA4 to synergistically activate the mouse Dio1 promoter, suggesting that Dio1 is regulated by thyroid hormone in the mouse through an indirect mechanism requiring prior KLF9 induction. In addition, we showed that physical interactions between the C-terminal zinc finger domain (Cf) of GATA4 and activation function 2 of HNF4alpha and between the basic domain adjacent to Cf of GATA4 and a C-terminal domain of KLF9 are both required for this synergistic response. Taken together, these results suggest that HNF4alpha regulates thyroid hormone homeostasis through transcriptional regulation of the mouse Dio1 gene with GATA4 and KLF9. PMID- 18426913 TI - Sphingosine kinases and sphingosine-1-phosphate are critical for transforming growth factor beta-induced extracellular signal-regulated kinase 1 and 2 activation and promotion of migration and invasion of esophageal cancer cells. AB - Transforming growth factor beta (TGFbeta) plays a dual role in oncogenesis, acting as both a tumor suppressor and a tumor promoter. These disparate processes of suppression and promotion are mediated primarily by Smad and non-Smad signaling, respectively. A central issue in understanding the role of TGFbeta in the progression of epithelial cancers is the elucidation of the mechanisms underlying activation of non-Smad signaling cascades. Because the potent lipid mediator sphingosine-1-phosphate (S1P) has been shown to transactivate the TGFbeta receptor and activate Smad3, we examined its role in TGFbeta activation of extracellular signal-regulated kinases 1 and 2 (ERK1/2) and promotion of migration and invasion of esophageal cancer cells. Both S1P and TGFbeta activate ERK1/2, but only TGFbeta activates Smad3. Both ligands promoted ERK1/2-dependent migration and invasion. Furthermore, TGFbeta rapidly increased S1P, which was required for TGFbeta-induced ERK1/2 activation, as well as migration and invasion, since downregulation of sphingosine kinases, the enzymes that produce S1P, inhibited these responses. Finally, our data demonstrate that TGFbeta activation of ERK1/2, as well as induction of migration and invasion, is mediated at least in part by ligation of the S1P receptor, S1PR2. Thus, these studies provide the first evidence that TGFbeta activation of sphingosine kinases and formation of S1P contribute to non-Smad signaling and could be important for progression of esophageal cancer. PMID- 18426915 TI - FANCJ helicase defective in Fanconia anemia and breast cancer unwinds G quadruplex DNA to defend genomic stability. AB - FANCJ mutations are associated with breast cancer and genetically linked to the bone marrow disease Fanconi anemia (FA). The genomic instability of FA-J mutant cells suggests that FANCJ helicase functions in the replicational stress response. A putative helicase with sequence similarity to FANCJ in Caenorhabditis elegans (DOG-1) and mouse (RTEL) is required for poly(G) tract maintenance, suggesting its involvement in the resolution of alternate DNA structures that impede replication. Under physiological conditions, guanine-rich sequences spontaneously assemble into four-stranded structures (G quadruplexes [G4]) that influence genomic stability. FANCJ unwound G4 DNA substrates in an ATPase dependent manner. FANCJ G4 unwinding is specific since another superfamily 2 helicase, RECQ1, failed to unwind all G4 substrates tested under conditions in which the helicase unwound duplex DNA. Replication protein A stimulated FANCJ G4 unwinding, whereas the mismatch repair complex MSH2/MSH6 inhibited this activity. FANCJ-depleted cells treated with the G4-interactive compound telomestatin displayed impaired proliferation and elevated levels of apoptosis and DNA damage compared to small interfering RNA control cells, suggesting that G4 DNA is a physiological substrate of FANCJ. Although the FA pathway has been classically described in terms of interstrand cross-link (ICL) repair, the cellular defects associated with FANCJ mutation extend beyond the reduced ability to repair ICLs and involve other types of DNA structural roadblocks to replication. PMID- 18426914 TI - Functional interaction between the Fab-7 and Fab-8 boundaries and the upstream promoter region in the Drosophila Abd-B gene. AB - Boundary elements have been found in the regulatory region of the Drosophila melanogaster Abdominal-B (Abd-B) gene, which is subdivided into a series of iab domains. The best-studied Fab-7 and Fab-8 boundaries flank the iab-7 enhancer and isolate it from the four promoters regulating Abd-B expression. Recently binding sites for the Drosophila homolog of the vertebrate insulator protein CTCF (dCTCF) were identified in the Fab-8 boundary and upstream of Abd-B promoter A, with no binding of CTCF to the Fab-7 boundary being detected either in vivo or in vitro. Taking into account the inability of the yeast GAL4 activator to stimulate the white promoter when its binding sites are separated by a 5-kb yellow gene, we have tested the functional interactions between the Fab-7 and Fab-8 boundaries and between these boundaries and the upstream promoter A region containing a dCTCF binding site. It has been found that dCTCF binding sites are essential for pairing between two Fab-8 insulators. However, a strong functional interaction between the Fab-7 and Fab-8 boundaries suggests that additional, as yet unidentified proteins are involved in long-distance interactions between them. We have also shown that Fab-7 and Fab-8 boundaries effectively interact with the upstream region of the Abd-B promoter. PMID- 18426916 TI - The anaphase-promoting complex/cyclosome controls repair and recombination by ubiquitylating Rhp54 in fission yeast. AB - Homologous recombination (HR) is important for maintaining genome integrity and for the process of meiotic chromosome segregation and the generation of variation. HR is regulated throughout the cell cycle, being prevalent in the S and G2 phases and suppressed in the G1 phase. Here we show that the anaphase promoting complex/cyclosome (APC/C) regulates homologous recombination in the fission yeast Schizosaccharomyces pombe by ubiquitylating Rhp54 (an ortholog of Rad54). We show that Rhp54 is a novel APC/C substrate that is destroyed in G1 phase in a KEN-box- and Ste9/Fizzy-related manner. The biological consequences of failing to temporally regulate HR via Rhp54 degradation are seen in haploid cells only in the absence of antirecombinase Srs2 function and are more extensive in diploid cells, which become sensitive to a range of DNA-damaging agents, including hydroxyurea, methyl methanesulfonate, bleomycin, and UV. During meiosis, expression of nondegradable Rhp54 inhibits interhomolog recombination and stimulates sister chromatid recombination. We thus propose that it is critical to control levels of Rhp54 in G1 to suppress HR repair of double-strand breaks and during meiosis to coordinate interhomolog recombination. PMID- 18426918 TI - miRDB: a microRNA target prediction and functional annotation database with a wiki interface. AB - MicroRNAs (miRNAs) are short noncoding RNAs that are involved in the regulation of thousands of gene targets. Recent studies indicate that miRNAs are likely to be master regulators of many important biological processes. Due to their functional importance, miRNAs are under intense study at present, and many studies have been published in recent years on miRNA functional characterization. The rapid accumulation of miRNA knowledge makes it challenging to properly organize and present miRNA function data. Although several miRNA functional databases have been developed recently, this remains a major bioinformatics challenge to miRNA research community. Here, we describe a new online database system, miRDB, on miRNA target prediction and functional annotation. Flexible web search interface was developed for the retrieval of target prediction results, which were generated with a new bioinformatics algorithm we developed recently. Unlike most other miRNA databases, miRNA functional annotations in miRDB are presented with a primary focus on mature miRNAs, which are the functional carriers of miRNA-mediated gene expression regulation. In addition, a wiki editing interface was established to allow anyone with Internet access to make contributions on miRNA functional annotation. This is a new attempt to develop an interactive community-annotated miRNA functional catalog. All data stored in miRDB are freely accessible at http://mirdb.org. PMID- 18426917 TI - How antigen quantity and quality determine T-cell decisions in lymphoid tissue. AB - T lymphocytes (T cells) express T-cell receptor (TCR) molecules on their surface that can recognize peptides (p) derived from antigenic proteins bound to products of the major histocompatibility complex (MHC) genes. The pMHC molecules are expressed on the surface of antigen-presenting cells, such as dendritic cells (DCs). T cells first encounter antigen on DCs in lymph nodes (LN). Intravital microscopy experiments show that upon entering the LN containing antigen, CD8+ T cells first move rapidly. After a few hours, they stop and make extended contacts with DCs. The factors that determine when and how this transition occurs are not well understood. We report results from computer simulations that suggest that the duration of phase one is related to the low probability of productive interactions between T cells and DCs. This is demonstrated by our finding that the antigen dose and type determine when such a transition occurs. These results are in agreement with experimental observations. TCR-pMHC binding characteristics and the antigen dose determine the time required for a productive T-cell-DC encounter (resulting in sustained contact). We find that the ratio of this time scale and the half-life of the pMHC complex itself provide a consolidated measure of antigen quantity and type. Results obtained upon varying different measures of antigen quantity and type fall on one curve when graphed against this ratio of time scales. Thus, we provide a mechanism for how the effects of varying one set of parameters are influenced by other prevailing conditions. This understanding should help guide future experimentation. PMID- 18426919 TI - A viral RNA competitively inhibits the antiviral endoribonuclease domain of RNase L. AB - Ribonuclease L (RNase L) is a latent endoribonuclease in an evolutionarily ancient interferon-regulated dsRNA-activated antiviral pathway. 2'-5' oligoadenylate (2-5A), the product of dsRNA-activated oligoadenylate synthetases (OASes), binds to ankyrin repeats near the amino terminus of RNase L, initiating a series of conformational changes that result in the activation of the endoribonuclease. A phylogenetically conserved RNA structure within group C enteroviruses inhibits the endoribonuclease activity of RNase L. In this study we report the mechanism by which group C enterovirus RNA inhibits RNase L. Viral RNA did not affect 2-5A binding to RNase L. Rather, the viral RNA inhibited the endoribonuclease domain. We used purified RNase L, purified 2-5A, and an RNA substrate with a 5' fluorophore and 3' quencher in FRET assays to measure inhibition of RNase L activity by the viral RNA. The group C enterovirus RNA was a competitive inhibitor of the endoribonuclease with a K(i) of 34 nM. Consistent with the kinetic profile of a competitive inhibitor, the viral RNA inhibited the constitutively active endoribonuclease domain of RNase L. We call this viral RNA the RNase L competitive inhibitor RNA (RNase L ciRNA). PMID- 18426920 TI - Selection and characterization of anti-NF-kappaB p65 RNA aptamers. AB - NF-kappaB transcription factors include a group of five mammalian proteins that form hetero- or homodimers and regulate hundreds of target genes involved in acute inflammation, HIV-1 transcription activation, and resistance to cancer therapy. We previously used in vitro selection to develop a small RNA aptamer (anti-p50) that binds the DNA-binding domain of NF-kappaB p50(2) with low nanomolar affinity but does not bind NF-kappaB p65(2). Here, we report the in vitro selection of anti-NF-kappaB p65 RNA aptamers using parallel in vitro selections with either a fully randomized RNA library or a degenerate RNA library based on the primary sequence of the 31-nucleotide anti-p50 RNA aptamer. We report the characterization of these aptamers with respect to NF-kappaB target specificity, affinity, minimal sequence requirements, secondary structure, and competition with DNA kappaB sites. These results expand opportunities for artificial inhibition of NF-kappaB transcription factor dimers containing p65 subunits. PMID- 18426921 TI - DcpS scavenger decapping enzyme can modulate pre-mRNA splicing. AB - The human scavenger decapping enzyme, DcpS, functions to hydrolyze the resulting cap structure following cytoplasmic mRNA decay yet is, surprisingly, a nuclear protein by immunofluorescence. Here, we show that DcpS is a nucleocytoplasmic shuttling protein that contains separable nuclear import and Crm-1-dependent export signals. We postulated that the presence of DcpS in both cellular compartments and its ability to hydrolyze cap structure may impact other cellular events dependent on cap-binding proteins. An shRNA-engineered cell line with markedly diminished DcpS levels led to a corresponding reduction in cap-proximal intron splicing of a reporter minigene and endogenous genes. The impaired cap catabolism and resultant imbalanced cap concentrations were postulated to sequester the cap-binding complex (CBC) from its normal splicing function. In support of this explanation, DcpS efficiently displaced the nuclear cap-binding protein Cbp20 from cap structure, and complementation with Cbp20 reversed the reduced splicing, indicating that modulation of splicing by DcpS is mediated through Cbp20. Our studies demonstrate that the significance of DcpS extends beyond its well-characterized role in mRNA decay and involves a broader range of functions in RNA processing including nuclear pre-mRNA splicing. PMID- 18426923 TI - Neonatal outcomes with caesarean delivery at term. PMID- 18426924 TI - Aplasia cutis in association with a triplet pregnancy and fetus papyraceus. PMID- 18426922 TI - Nucleoside modifications modulate activation of the protein kinase PKR in an RNA structure-specific manner. AB - The human interferon-induced protein kinase PKR is a key component of innate immunity, a process in which it senses pathogenic RNA. PKR consists of an N terminal dsRNA-binding domain (dsRBD) and a C-terminal kinase domain. Upon binding long (>33 base pairs) stretches of pathogenic dsRNA, PKR undergoes autophosphorylation, which activates it to phosphorylate eIF2alpha, leading to inhibition of translation initiation. Many cellular and viral transcripts contain nucleoside modifications, and these could affect PKR activation. For example, a 5'-triphosphate confers the ability of relatively unstructured transcripts to activate PKR. Effects of internal RNA modifications on PKR activation have not been reported. Herein, PKR activation by ssRNA and dsRNA containing internal nucleobase, sugar, and phosphodiester modifications is analyzed. We find that for 5'-triphosphate-containing ssRNA, most base and sugar modifications abrogate activation, although 2'-fluoro-modified ssRNA does not, indicative of a critical role for hydrogen bonding at the ribose sugar. In the case of dsRNA, a more limited set of nucleoside modifications affect PKR activation. Watson-Crick base pairing is required for activation, and some minor groove modifications abrogate activation while major groove modifications have little effect. Surprisingly, GU wobble pairs also largely abrogate dsRNA-mediated activation when present at modest levels. Modifications to dsRNA that abrogate activation have no significant effect on dsRBD binding, allowing such RNAs to act as inhibitors and suggesting a nonequivalence of binding ability and activation. Overall, the findings indicate that nucleoside modifications and wobble pairing may serve to discriminate self-RNA and pathogenic RNA in innate immunity. PMID- 18426925 TI - Kneeling delivery in America 2000 years ago. PMID- 18426926 TI - Dr Leonard Colebrook, FRS (1883-1967) and the chemotherapeutic conquest of puerperal infection. PMID- 18426927 TI - Meta-variability of advice on drugs in the breastfeeding mother: the example of beta-blockers. PMID- 18426928 TI - Group B streptococcus and preventive strategies in Europe. PMID- 18426929 TI - Need to address all forms of childhood malnutrition with a common agenda. PMID- 18426930 TI - Children affected by domestic abuse while abroad on holiday. PMID- 18426931 TI - Head-injury-induced pituitary dysfunction. An old curiosity rediscovered. PMID- 18426934 TI - Erythema ab igne and Crohn's disease. PMID- 18426936 TI - Maternal smoking habits are associated with differences in infants' long-chain polyunsaturated fatty acids in whole blood: a case-control study. AB - OBJECTIVE: To study the effects of maternal smoking on the status of long-chain polyunsaturated fatty acids (LCPUFA) in infants' whole-blood lipids. DESIGN: A case-control matched study planned on the basis of preliminary observations. SETTING: Maternity ward. PATIENTS: A total of 159 healthy, term, breastfed infants with weight appropriate for gestational age, subdivided (53 per group) into those born to non-smokers (reference), smokers (> or = 5 cigarettes per day) who either stopped within the first trimester of pregnancy (early smokers) or who continued througout pregnancy (late smokers). INTERVENTIONS: The fatty acid profile of 4-day-old infants was determined on whole blood. RESULTS: Higher levels of linoleic (LA) and alpha-linolenic acid (ALA) and lower levels of the metabolic products di-homo-gammalinolenic (DHGLA) and arachidonic (AA), of the n 6 series, and docosahexaenoic acid (DHA), of the n-3 series, were found in infants born to late smokers compared with the reference group. The DHGLA/LA and AA/DHGLA ratios in the n-6 series and DHA/ALA in the n-3 series, which are indices of the metabolic processes in LCPUFA synthesis, were lower in infants born to smokers compared with those born to non-smokers. Infants born to early smokers showed n-6 PUFA levels and ratios similar to references and n-3 parameters closer to those born to late smokers. No dietary differences were found among the three groups of mothers. All the independent associations with smoking persisted after adjustment for maternal covariates. Pre-pregnancy body weight, which is lower in late smokers compared with non-smokers, independently correlated with LCPUFA levels in both series. CONCLUSIONS: Maternal smoking is associated with a reduction in LCPUFA pools in infants, which might have structural and functional consequences. PMID- 18426938 TI - Jole Rider and bicycles for Africa. PMID- 18426939 TI - Intrabiliary rupture of hydatid cyst demonstrated by magnetic resonance cholangiopancreatography. PMID- 18426941 TI - Internet access, utilisation and perception by parents. PMID- 18426942 TI - Management of infantile spasms in a regional centre before and after the United Kingdom infantile spasms study (UKISS). PMID- 18426943 TI - Sitting devices could protect against sudden infant death. PMID- 18426944 TI - Vaccine-related pain: picking up the child soon after the injection reduces the time of crying. PMID- 18426945 TI - Parents' reactions to nose bleeds and salt ingestion in infancy. PMID- 18426946 TI - Which is the better marker for susceptibility to disease later in life--low birthweight or prematurity? PMID- 18426947 TI - Pitfalls in the diagnosis and management of transient synovitis of the hip: a retrospective case-note analysis. PMID- 18426948 TI - Resuscitate with the placental circulation intact. PMID- 18426949 TI - State of the world's children and progress towards the Alma Ata Declaration. PMID- 18426950 TI - Nasal nitric oxide for early diagnosis of familial primary ciliary dyskinesia. PMID- 18426951 TI - Prevalence of anaemia in an inner city primary school population. PMID- 18426953 TI - Rate-limiting steps in hepatic drug clearance: comparison of hepatocellular uptake and metabolism with microsomal metabolism of saquinavir, nelfinavir, and ritonavir. AB - The intrinsic metabolic clearance of saquinavir, nelfinavir, and ritonavir was determined over a range of concentrations (0.02-20 microM) in both rat liver microsomes and fresh isolated rat hepatocytes in suspension. Clearance values were found to be concentration dependent for both systems, and at low concentrations, microsomal clearance was much greater (7-14-fold) than in hepatocytes. Kinetic parameters showed substantially lower microsomal K(m) values (5-42 nM) compared with suspended rat hepatocytes (34-270 nM) but similar scaled V(max) values (2-26 nmol/min/g liver). In the absence of metabolism (achieved by pretreating hepatocytes with a mechanism-based inhibitor of cytochrome P450), saquinavir, nelfinavir, and ritonavir were actively and rapidly taken up into hepatocytes (cell/medium concentration ratios of 306-3352), and intracellular unbound drug concentrations between 5- and 12-fold higher than extracellular unbound concentrations were achieved. Comparison of the rate of uptake into hepatocytes with the rate of metabolism in hepatocytes and microsomes indicates that the former is the rate-limiting step at low concentrations. The rate of metabolism saturates at lower concentrations (100-400-fold) than the rate of uptake; hence, at the high concentrations metabolic rate-limited clearance occurs. In conclusion, the clearance of saquinavir, nelfinavir, and ritonavir is extremely rapid, and it is proposed that in the case of hepatocytes and by inference in vivo, the rate of uptake limits the metabolic clearance of these three drugs. PMID- 18426954 TI - Trend analysis of a database of intravenous pharmacokinetic parameters in humans for 670 drug compounds. AB - We present herein a compilation and trend analysis of human i.v. pharmacokinetic data on 670 drugs representing, to our knowledge, the largest publicly available set of human clinical pharmacokinetic data. This data set provides the drug metabolism scientist with a robust and accurate resource suitable for a number of applications, including in silico modeling, in vitro-in vivo scaling, and physiologically based pharmacokinetic approaches. Clearance, volume of distribution at steady state, mean residence time, and terminal phase half-life were obtained or derived from original references exclusively from studies utilizing i.v. administration. Plasma protein binding data were collected from other sources to supplement these pharmacokinetic data. These parameters were analyzed concurrently with a range of simple physicochemical descriptors, and resultant trends and patterns within the data are presented. Our findings with this much expanded data set were consistent with earlier described notions of trends between physicochemical properties and pharmacokinetic behavior. These observations and analyses, along with the large database of human pharmacokinetic data, should enable future efforts aimed toward developing quantitative structure pharmacokinetic relationships and improving our understanding of the relationship between fundamental chemical characteristics and drug disposition. PMID- 18426955 TI - Prediction of the pharmacokinetics of atorvastatin, cerivastatin, and indomethacin using kinetic models applied to isolated rat hepatocytes. AB - The disposition of atorvastatin, cerivastatin, and indomethacin, established substrates of rat hepatic basolateral uptake transporters, has been evaluated in suspended rat hepatocytes. Cell and media concentration-time data were simultaneously fitted to a model incorporating active uptake, permeation, binding, and metabolism. Use of the model to estimate the ratio of intracellular to extracellular steady-state free drug concentrations demonstrated the strong influence of active uptake on the kinetics of atorvastatin (18:1) and cerivastatin (8:1), in comparison with indomethacin (3.5:1). Indomethacin, however, was shown to have a higher uptake clearance (599 +/- 101 microl/min/10(6) cells) than atorvastatin (375 +/- 45 microl/min/10(6) cells) and cerivastatin (413 +/- 47 microl/min/10(6) cells). The high passive permeability of indomethacin (237 +/- 63 microl/min/10(6) cells) clearly negated the effect of the active transport on the overall disposition. An analogous physiological model was constructed that allowed prediction of the in vivo pharmacokinetics, including the free intracellular concentration in liver. Hepatic clearance was well predicted by the model, in contrast to predictions based on standard methods. Volume of distribution was well predicted for indomethacin and predicted reasonably for atorvastatin and cerivastatin and higher than might be expected for an acid compound. Furthermore, the terminal half-life predictions for all three compounds were within 2-fold of the observed values. The ability to estimate the free-intracellular hepatic concentration of uptake substrates has major benefits in terms of predicting pharmacokinetics, potential CYP-mediated drug-drug interactions, and efficacy of hepatically targeted therapeutics. PMID- 18426956 TI - The biological sciences section program at the 60th Annual Meeting of the Gerontological Society of America. AB - In this era of genomics and other exciting technical advances, research on the biology of aging is undergoing a renaissance. This report summarizes 10 cutting edge areas of research covered in symposia that spanned such topics as stem cells, novel vaccine strategies, nutritional sensing, new concepts of Parkinson's disease, high throughput screening for aging interventions, manipulating telomerase in cancer and immunodeficiency, synergy between aging and HIV disease, and epigenetic influences on aging. Novel animal models, including those showing no evidence of aging, as well as ethical and political implications of embryonic stem cells and alternative medicine are also discussed. PMID- 18426957 TI - They really are that old: a validation study of centenarian prevalence in Okinawa. AB - Long-lived individuals, such as centenarians, may harbor genetic or environmental advantages important for healthy aging. Populations with high prevalence of such individuals offer fertile ground for such research. However, precise phenotypes are required, particularly accurate age. Okinawa has among the world's highest reported prevalence of centenarians but, despite extensive study, no systematic validation of centenarian prevalence has been published. Therefore, we performed comprehensive age validation of a subset (8%) of the total centenarian population and assessed the reliability of the age registration system. Self-reported age was validated with several common methods and found to correlate well with documented age. Demographic methods, including assessment of age heaping, maximum age at death, centenarian proportions, and male to female ratios of centenarians indicate that the age registration system is reliable. We conclude that the high reported centenarian prevalence in Okinawa is valid and warrants further study for its genetic and environmental correlates. PMID- 18426958 TI - Age-induced morphological, biochemical, and functional alterations in isolated mitochondria from murine skeletal muscle. AB - Several in vitro studies about age-associated skeletal muscle mitochondrial dysfunction are somewhat conflicting, and this might be related to different normalization procedures. The objective of this study was to normalize the functional and biochemical data per number of mitochondria present in a mitochondrial suspension. Functional and biochemical parameters were obtained in mitochondrial suspensions from murine skeletal muscle of different ages. Mitochondrial respiratory function was polarographically measured using a Clark type oxygen electrode. Biochemical analyses included determination of citrate synthase (CS) activity and total protein content in the mitochondrial suspension. Electron microscopy analysis of the suspensions allowed calculation of the number of mitochondria per milligram of protein. Our results conclude that advanced age is associated with mitochondrial dysfunction; moreover, from the correlation between morphological and biochemical data, it is evident that CS activity in the mitochondrial suspensions is a more accurate marker of mitochondrial mass than is total protein content. PMID- 18426959 TI - Age-related correlation between antioxidant enzymes and DNA damage with smoking and body mass index. AB - To understand whether oxidants contribute to the initiation and/or promulgation toward aging, the present study has been undertaken on 220 healthy male volunteers aged 20-80 years selected from the defined electoral area (suburbs of Tirupati, Andhra Pradesh, India) to evaluate the concentrations of free radicals (superoxide anion, hydrogen peroxide), lymphocyte antioxidant enzymes (glutathione S-transferase, superoxide dismutase, catalase), and DNA damage in relation to obesity and smoking (lifestyles). A two fold increase of lymphocyte free radical generation (DNA damage) was observed in older age groups with a reduced antioxidant potential, forming a link between cigarette smoking and oxidative stress represented by an antioxidant imbalance. Body mass index had a positive relationship with oxidative stress, but antioxidant levels did not vary with body mass index. The findings conclude that free radical-mediated oxidative stress and DNA damage accelerate with lifestyle variations under reduced antioxidant potential. PMID- 18426961 TI - Low plasma carotenoids and skeletal muscle strength decline over 6 years. AB - BACKGROUND: Higher intake of fruits and vegetables appears to protect against inflammation, poor physical performance, and disability, but its relationship with muscle strength is unclear. We examined the association between total plasma carotenoids, an indicator of fruit and vegetable intake, and changes in muscle strength over a 6-year follow-up in the participants aged 65 years and older in the InCHIANTI study, a population-based study in Tuscany, Italy. METHODS: Plasma carotenoids were measured at enrollment (1998-2000). Hip, knee, and grip strength were measured at enrollment and 6 years later (2004-2006) in 628 of the 948 participants evaluated at baseline. Poor muscle strength was defined as the lowest sex-specific quartile of hip, knee, and grip strength at enrollment. The main outcome was poor muscle strength at the 6-year follow-up visit among those participants originally in the upper three quartiles of strength at enrollment. RESULTS: Overall, 24.9% (110/441), 25.0% (111/444), and 24.9% (118/474) participants developed poor hip, knee, and grip strength, respectively. After adjusting for potential confounders, participants in the lowest versus the highest quartile of total plasma carotenoids at enrollment were at higher risk of developing poor hip (odds ratio [OR] = 3.01, 95% CI, 1.43-6.31, p =.004), knee (OR = 2.89, 95% CI, 1.38-6.02, p =.005), and grip (OR = 1.88, 95% CI, 0.93-3.56, p =.07) muscle strength at the 6-year follow-up visit. CONCLUSION: These findings suggest that older community-dwelling adults with lower plasma carotenoids levels, a marker of poor fruit and vegetable intake, are at a higher risk of decline in skeletal muscle strength over time. PMID- 18426960 TI - Statin use and incident frailty in women aged 65 years or older: prospective findings from the Women's Health Initiative Observational Study. AB - BACKGROUND: Inflammatory biomarkers have shown consistent associations with disability and frailty in older adults. Statin medications may reduce the incidence the frailty because of their anti-inflammatory effects. This study examines associations between current use, duration, and potency of statin medications and incident frailty in initially nonfrail women 65 years old or older. METHODS: The authors conducted a prospective analysis of data from the Women's Health Initiative Observational Study (WHI-OS) conducted at 40 clinical centers in the United States. Eligible women were nonfrail and 65-79 years old at baseline (n = 25,378). Current statin use at baseline was ascertained through direct inspection of medicine containers during clinic visits. Frailty was ascertained through self-reported indicators and physical measurements at baseline and 3-year clinic contacts. Components of frailty included self-reported low physical function, exhaustion, low physical activity, and unintended weight loss. Multinomial logistic regression models were used to adjust for covariates predicting incident frailty. RESULTS: Among the 25,378 eligible women, 3453 (13.6%) developed frailty by the 3-year follow-up contact. Current statin use had no association with incident frailty (multivariate-adjusted odds ratio [OR] = 1.00; 95% confidence interval [CI], 0.85-1.16). Duration and potency of statin use were also not significantly associated with incident frailty. Among low potency statin users, longer duration of use was associated with reduced risk of frailty (p for trend =.02). A similar pattern of results was observed when frailty was studied in the absence of intervening, incident cardiovascular events. CONCLUSIONS: Overall, incidence of frailty was similar in current statin users and nonusers. PMID- 18426962 TI - The relation of peripheral arterial disease to leg force, gait speed, and functional dependence among older adults. AB - BACKGROUND: Atherosclerotic peripheral arterial disease (PAD), common among older adults, is associated with poor low-extremity functioning. In considering functional status, varying domains exist, including activities of daily living (ADL), instrumental activities of daily living (IADL), low-extremity mobility (LEM), and leisure and/or social activities (LSA). However, little is known about how PAD is related to functional status beyond low-extremity functioning. METHODS: A total of 1798 participants 60 years old or older was selected from the population-based National Health and Nutrition Examination Survey 1999-2002 in the United States. ADL, IADL, LSA, LEM, and general physical activities (GPA) were obtained by self-report. Peak leg force was obtained from an isokinetic dynamometer. Habitual gait speed was obtained from a 20-foot timed walk. PAD was defined as an ankle-brachial blood pressure index <0.9 in either leg. RESULTS: After multivariable adjustment, the odds ratios (ORs) for dependence in IADL, LSA, and LEM comparing participants with PAD to those without were 1.60 (95% confidence interval [CI], 1.11-2.29), 1.63 (95% CI, 1.08-2.44), and 2.29 (95% CI, 1.64-3.18), respectively. Additional adjustment of peak leg force and/or habitual gait speed diminished the relations of PAD to dependence in IADL and LSA. PAD was associated with an 18.06 Newton reduction (p =.003) in peak leg force and a 0.05 m/s reduction (p =.002) in habitual gait speed. CONCLUSION: PAD was independently associated with multiple domains of functional dependence. The association between PAD and dependence in IADL and LSA was to a large extent mediated by leg force and gait speed. PMID- 18426963 TI - The physical and biological characterization of a frail mouse model. AB - BACKGROUND: The development of animal models that approximate human frailty is necessary to facilitate etiologic and treatment-focused frailty research. The genetically altered IL-10(tm/tm) mouse does not express the antiinflammatory cytokine interleukin 10 (IL-10) and is, like frail humans, more susceptible to inflammatory pathway activation. We hypothesized that with increasing age, IL 10(tm/tm) mice would develop physical and biological characteristics similar to those of human frailty as compared to C57BL/6J control mice. METHODS: Strength, activity, serum IL-6, and skeletal muscle gene expression were compared between age-matched and gender-matched IL-10(tm/tm) mice on C57BL/6J background and C57BL/6J control mice using a longitudinal design for physical characteristics and cross-sectional design for biological characteristics. RESULTS: Strength levels declined significantly faster in IL-10(tm/tm) compared to control mice with increasing age. Serum IL-6 levels were significantly higher in older compared to younger IL-10(tm/tm) mice and were significantly higher in older IL 10(tm/tm) compared to age- and gender-matched C57BL/6J control mice. One hundred twenty-five genes, many related to mitochondrial biology and apoptosis, were differentially expressed in skeletal muscle between 50-week-old IL-10(tm/tm) and 50-week-old C57BL/6J mice. No expression differences between IL-10(tm/tm) age groups were identified by quantitative polymerase chain reaction. CONCLUSION: These physical and biological findings suggest that the IL-10(tm/tm) mouse develops inflammation and strength decline consistent with human frailty at an earlier age compared to C57BL/6J control type mice. This finding provides rationale for the further development and utilization of the IL-10(tm/tm) mouse to study the biological basis of frailty. PMID- 18426964 TI - Caregiving in the presence of chronic pain. AB - BACKGROUND: Chronic pain sufferers and caregivers share the risk of higher levels of psychological distress and adverse effects on well-being. This study examined the joint impact of chronic pain and primary caregiving on older people. METHODS: Data came from the New South Wales (NSW) Older People's Health Survey 1999, a state-wide general health survey of over 9000 NSW residents 65 years old or older. Using survey logistic regression modeling, we examined the relationship between chronic pain with different levels of disability, caregiving status, self reported physical functioning, and two dependent variables--poor/fair self-rated health and psychological distress. RESULTS: Caregivers with chronic pain reported more psychological distress and poorer self-rated health than caregivers without pain, when both were compared to noncaregivers without pain (age-adjusted and sex adjusted odds ratios [ORs] for caregivers with pain were 3.4 and 2.8, respectively, both p <.001). Caregivers with pain and noncaregivers with pain had similar patterns of results. Physical function significantly declined for both caregivers and noncaregivers with pain when compared with noncaregivers without pain. CONCLUSIONS: Older people coping with caregiving and chronic pain are a potentially vulnerable group. Chronic pain status should be ascertained in older people who are caregivers, with particular attention to the issue of caregiver psychological distress and physical well-being. PMID- 18426965 TI - Extrinsic risk factors for pressure ulcers early in the hospital stay: a nested case-control study. AB - BACKGROUND: Little is known about the impact of extrinsic factors on pressure ulcer risk. The objective of this study was to determine whether risk of pressure ulcers early in the hospital stay is associated with extrinsic factors such as longer emergency department (ED) stays, night or weekend admission, potentially immobilizing procedures and medications, and admission to an intensive care unit (ICU). METHODS: A nested case-control study was performed in two teaching hospitals in Philadelphia, Pennsylvania. Participants were medical patients age > or =65 years admitted through the ED. Cases (n = 195) had > or =1 possibly or definitely hospital-acquired pressure ulcers. Three controls per case were sampled randomly from among noncases at the same hospital in the same month (n = 597). Pressure ulcer status was determined by a research nurse on the third day of hospitalization. Pressure ulcers were classified as preexisting, possibly hospital-acquired, or definitely hospital-acquired. Information on extrinsic factors was obtained by chart review. RESULTS: The odds of pressure ulcers were twice as high for those with an ICU stay as for those without (adjusted odds ratio [aOR] 2.0, 95% confidence interval [CI], 1.2-3.5). The aOR was 0.6 (95% CI, 0.3-0.9) for use of any potentially immobilizing medications during the early inpatient period. CONCLUSIONS: Many of the procedures experienced by patients in the ED and early in the inpatient stay do not confer excess pressure ulcer risk. Having an ICU stay is associated with a doubling of risk. This finding emphasizes the importance of developing and evaluating interventions to prevent pressure ulcers among patients in the ICU. PMID- 18426966 TI - The metabolic syndrome is associated with circulating adipokines in older adults across a wide range of adiposity. AB - BACKGROUND: Circulating levels of adipokines are elevated with adiposity and are closely linked with the clustering of traditional metabolic risk factors for cardiovascular disease. The purpose of this study was to examine the relationship of metabolic syndrome to several adipokines and the role of total and visceral adiposity in influencing this relationship in older adults. METHODS: A cross sectional analysis was conducted including 1914 individuals aged 70-79 years without cardiovascular disease or type 2 diabetes. The metabolic syndrome was defined by the updated Adult Treatment Panel III criteria. Circulating levels of leptin, adiponectin, plasminogen activator inhibitor type 1 (PAI-1), interleukin 6 (IL-6), tumor necrosis factor-alpha (TNF-alpha), and C-reactive protein (CRP) were measured. Body composition and abdominal visceral fat area were determined. RESULTS: Both the presence of metabolic syndrome and the number of metabolic syndrome components were associated with higher levels of leptin, PAI-1, IL-6, TNF-alpha, and CRP and with lower levels of adiponectin (all p <.0001). The odds ratios for the prevalence of metabolic syndrome associated with adipokines were attenuated after adjustment for total fat mass and/or visceral fat area, but remained significant. Levels of leptin, PAI-1, IL-6, and TNF-alpha were higher (all p <.05 to p <.0001), and adiponectin was lower (all p <.0001), in persons with, compared to those without, metabolic syndrome within each tertile of percent body fat. CONCLUSION: The metabolic syndrome is associated with adipokines in older adults across a wide range of adiposity, including in those with low or normal overall fatness. PMID- 18426967 TI - The white-black disability gap revisited: does an incident heart attack change this gap? AB - BACKGROUND: A myocardial infarction (MI) results typically in abrupt functional deterioration immediately postevent, followed by recovery. The post-MI health disparities experienced by black older adults may be attributable to the social and health correlates of race. We explored patterns of change in functional status in a community-based sample of 243 older white and black persons hospitalized for an incident MI. METHODS: The study sample was drawn from the Established Populations for Epidemiologic Studies of the Elderly (EPESE). All older adults hospitalized for an incident MI between the first two waves of data collection were followed up yearly for two additional years. Nonlinear quadratic trajectories of functional status, as measured by disability in activities of daily living (ADL) and functional limitation (FL), were fit using mixed-effects models. RESULTS: Although there were no nonlinear differences in ADL trajectories, there was a faster nonlinear rate of change in FL in older blacks compared to whites, independent of other social and health factors. The baseline white-black gap in FL widened after the MI by the first follow-up, continued to widen at a less accelerated pace until the second follow-up, and narrowed by the third follow-up. CONCLUSIONS: Disparities in relevant social and health factors did not account for the more abrupt deterioration in FL postevent or for the more substantial recovery in older blacks compared to older whites. Disparities in therapeutic strategies and the "survival of the fittest" may underlie the pattern of this white-black gap in FL after an incident MI. PMID- 18426968 TI - A short version of the ADAM Questionnaire for androgen deficiency in Chinese men. AB - BACKGROUND: A 10-question screening questionnaire for androgen deficiency in aging men (ADAM) was reported in previous white but not Chinese populations. We therefore investigated the validity of a Chinese version of the Saint Louis University ADAM questionnaire to screen for androgen deficiency in Chinese men. METHODS: This was a cross-sectional study. Seven hundred ninety-six ambulatory community-based Chinese men, 18-89 years old, were recruited from October 2003 through June 2006. Self-administered Chinese ADAM questionnaire and morning blood samples for serum total testosterone (TT) and bioavailable testosterone (BT) levels were collected from all participants. Low serum BT levels (androgen deficiency) were defined as <5th percentile of serum BT levels in young healthy Chinese men (18-29 years). RESULTS: The Chinese ADAM questionnaire had good internal consistency (Cronbach alpha = 0.74) and test-retest reliability (Pearson correlation coefficient, r = 0.86; p <.001, two-tailed). As a screening test for low serum BT levels, the Chinese ADAM questionnaire has a high sensitivity of 88% but low specificity of 32%. In 6 of the 10 questions, the mean serum BT levels were significantly lower in those who answered positively than in those who answered negatively. Using a cut-off score of > or =2, a six-question short Chinese ADAM questionnaire demonstrated sensitivity, specificity, and positive and negative predictive values of 86%, 40%, 46%, and 82%, respectively. CONCLUSION: We have validated a full Chinese version and developed a shortened version of the ADAM questionnaire, and demonstrated that they are sensitive but not specific screening tests for androgen deficiency in Chinese men. PMID- 18426969 TI - Abdominal applications of 3.0-T MR imaging: comparative review versus a 1.5-T system. AB - With the development of dedicated receiver coils and increased gradient performance, 3.0-T magnetic resonance (MR) systems are gaining wider acceptance in clinical practice. The expected twofold increase in signal-to-noise ratio (SNR) compared with that of 1.5-T MR systems may help improve spatial resolution or increase temporal resolution when used with parallel acquisition techniques. Several issues must be considered when applying 3.0-T MR in the abdomen, including the alteration of the radiofrequency field and relaxation time, increase in energy deposition and susceptibility effects, and problems associated with motion artifacts. For the evaluation of liver lesions, higher SNR and greater resolution achieved with the 3.0-T system could translate into better detection of malignant lesions on T2-weighted images obtained with adjusted imaging parameters. For the evaluation of pancreatic and biliary diseases, high resolution T2-weighted imaging using single-shot turbo spin-echo sequences is useful; improvement in SNR was noticeable on two-dimensional MR cholangiopancreatographic images. For the preoperative imaging of rectal cancer, a single-shot sequence is useful for dramatically decreasing imaging time while maintaining image quality. Substantial modification of examination protocols, with optimized imaging parameters and sequence designs along with ongoing development of hardware, could contribute to an increased role of the 3.0-T system for abdominal MR examinations. PMID- 18426970 TI - Differential roles of ICAM-1 and VCAM-1 in leukocyte-endothelial cell interactions in skin and brain of MRL/faslpr mice. AB - MRL/fas(lpr) mice, which undergo a systemic autoimmune disease with similarities to systemic lupus erythematosus (SLE), display reduced pathology and prolonged survival if rendered deficient in ICAM-1. However, it remains unclear whether this is a result of the ability of ICAM-1 to promote the immune response or mediate leukocyte recruitment. Therefore, the aim of these studies was to compare the role of ICAM-1 in the elevated leukocyte-endothelial interactions, which affect MRL/fas(lpr) mice. Intravital microscopy was used to compare leukocyte rolling and adhesion in postcapillary venules in the dermal and cerebral (pial) microcirculations of wild-type (ICAM+/+) and ICAM-1-deficient (ICAM-1-/-) MRL/fas(lpr) mice. In the dermal microcirculation of 16-week MRL/fas(lpr) mice, leukocyte adhesion was increased relative to nondiseased MRL+/+ mice. However, this increase was abolished in ICAM-1-/- MRL/fas(lpr) mice. ICAM-1 deficiency was also associated with reduced dermal pathology. In contrast, in the pial microcirculation, the elevation in leukocyte adhesion observed in ICAM+/+ MRL/fas(lpr) mice also occurred in ICAM-1-/- MRL/fas(lpr) mice. VCAM-1 expression was detectable in both vascular beds, but higher levels were detected in the pial vasculature. Furthermore, VCAM-1 blockade significantly reduced leukocyte adhesion and rolling in the cerebral microcirculation of ICAM-1-/- MRL/fas(lpr) mice. Therefore, ICAM-1 was critical for leukocyte adhesion in the skin but not the brain, where VCAM-1 assumed the major function. Given the ongoing development of anti-adhesion molecule therapies and their potential in inflammatory diseases such as SLE, these data indicate that implementation of these therapies in SLE should take into account the potential for tissue-specific functions of adhesion molecules. PMID- 18426971 TI - Type 1-polarized dendritic cells loaded with autologous tumor are a potent immunogen against chronic lymphocytic leukemia. AB - Induction of active tumor-specific immunity in patients with chronic lymphocytic leukemia (CLL) and other hematologic malignancies is compromised by the deficit of endogenous dendritic cells (DCs). In attempt to develop improved vaccination strategies for patients with CLL and other tumors with poorly identified rejection antigens, we tested the ability of ex vivo-generated DCs to cross present the antigens expressed by CLL cells and to induce CLL-specific, functional CTL responses. Monocyte-derived DCs from CLL patients were induced to mature using a "standard" cytokine cocktail (in IL-1beta, TNF-alpha, IL-6, and PGE2) or using an alpha-type 1-polarized DC (alphaDC1) cocktail (in IL-1beta, TNF alpha, IFN-alpha, IFN-gamma, and polyinosinic:polycytidylic acid) and were loaded with gamma-irradiated, autologous CLL cells. alphaDC1 from CLL patients expressed substantially higher levels of multiple costimulatory molecules (CD83, CD86, CD80, CD11c, and CD40) than standard DCs (sDCs) and immature DCs, and their expression of CCR7 showed intermediate level. alphaDC1 secreted substantially higher (10-60 times) levels of IL-12p70 than sDCs. Although alphaDC1 and sDCs showed similar uptake of CLL cells, alphaDC1 induced much higher numbers (range, 2.4-38 times) of functional CD8+ T cells against CLL cells. The current demonstration that autologous tumor-loaded alphaDC1 are potent inducers of CLL specific T cells helps to develop improved immunotherapies of CLL. PMID- 18426972 TI - Sir John Gurdon: godfather of cloning. Interviewed by Ruth Williams. PMID- 18426973 TI - The peroxisome: a production in four acts. AB - A cell regulates the number, size, and kind of each organelle it possesses in response to its particular role in an environment or tissue. Yet we still know little about how the molecular signaling networks within each cell perform such regulation. In this issue, Saleem et al. (Saleem, R.A., B. Knoblach, F.D. Mast, J.J. Smith, J. Boyle, C.M. Dobson, R. Long-O'Donnell, R.A. Rachubinski, and J.D. Aitchison. 2008. J. Cell Biol. 181:281-292) show for the first time how groups of kinases and phosphatases are organized to control when and how a cell assembles one kind of organelle, the peroxisome. PMID- 18426974 TI - Aurora B kinase and protein phosphatase 1 have opposing roles in modulating kinetochore assembly. AB - The outer kinetochore binds microtubules to control chromosome movement. Outer kinetochore assembly is restricted to mitosis, whereas the inner kinetochore remains tethered to centromeres throughout the cell cycle. The cues that regulate this transient assembly are unknown. We find that inhibition of Aurora B kinase significantly reduces outer kinetochore assembly in Xenopus laevis and human tissue culture cells, frog egg extracts, and budding yeast. In X. leavis M phase extracts, preassembled kinetochores disassemble after inhibiting Aurora B activity with either drugs or antibodies. Kinetochore disassembly, induced by Aurora B inhibition, is rescued by restraining protein phosphatase 1 (PP1) activity. PP1 is necessary for kinetochores to disassemble at the exit from M phase, and purified enzyme is sufficient to cause disassembly on isolated mitotic nuclei. These data demonstrate that Aurora B activity is required for kinetochore maintenance and that PP1 is necessary and sufficient to disassemble kinetochores. We suggest that Aurora B and PP1 coordinate cell cycle-dependent changes in kinetochore assembly though phosphorylation of kinetochore substrates. PMID- 18426975 TI - The highly conserved nuclear lamin Ig-fold binds to PCNA: its role in DNA replication. AB - This study provides insights into the role of nuclear lamins in DNA replication. Our data demonstrate that the Ig-fold motif located in the lamin C terminus binds directly to proliferating cell nuclear antigen (PCNA), the processivity factor necessary for the chain elongation phase of DNA replication. We find that the introduction of a mutation in the Ig-fold, which alters its structure and causes human muscular dystrophy, inhibits PCNA binding. Studies of nuclear assembly and DNA replication show that lamins, PCNA, and chromatin are closely associated in situ. Exposure of replicating nuclei to an excess of the lamin domain containing the Ig-fold inhibits DNA replication in a concentration-dependent fashion. This inhibitory effect is significantly diminished in nuclei exposed to the same domain bearing the Ig-fold mutation. Using the crystal structures of the lamin Ig fold and PCNA, molecular docking simulations suggest probable interaction sites. These findings also provide insights into the mechanisms underlying the numerous disease-causing mutations located within the lamin Ig-fold. PMID- 18426976 TI - Genome-wide analysis of signaling networks regulating fatty acid-induced gene expression and organelle biogenesis. AB - Reversible phosphorylation is the most common posttranslational modification used in the regulation of cellular processes. This study of phosphatases and kinases required for peroxisome biogenesis is the first genome-wide analysis of phosphorylation events controlling organelle biogenesis. We evaluate signaling molecule deletion strains of the yeast Saccharomyces cerevisiae for presence of a green fluorescent protein chimera of peroxisomal thiolase, formation of peroxisomes, and peroxisome functionality. We find that distinct signaling networks involving glucose-mediated gene repression, derepression, oleate mediated induction, and peroxisome formation promote stages of the biogenesis pathway. Additionally, separate classes of signaling proteins are responsible for the regulation of peroxisome number and size. These signaling networks specify the requirements of early and late events of peroxisome biogenesis. Among the numerous signaling proteins involved, Pho85p is exceptional, with functional involvements in both gene expression and peroxisome formation. Our study represents the first global study of signaling networks regulating the biogenesis of an organelle. PMID- 18426977 TI - eIF4GI links nutrient sensing by mTOR to cell proliferation and inhibition of autophagy. AB - Translation initiation factors have complex functions in cells that are not yet understood. We show that depletion of initiation factor eIF4GI only modestly reduces overall protein synthesis in cells, but phenocopies nutrient starvation or inhibition of protein kinase mTOR, a key nutrient sensor. eIF4GI depletion impairs cell proliferation, bioenergetics, and mitochondrial activity, thereby promoting autophagy. Translation of mRNAs involved in cell growth, proliferation, and bioenergetics were selectively inhibited by reduction of eIF4GI, as was the mRNA encoding Skp2 that inhibits p27, whereas catabolic pathway factors were increased. Depletion or overexpression of other eIF4G family members did not recapitulate these results. The majority of mRNAs that were translationally impaired with eIF4GI depletion were excluded from polyribosomes due to the presence of multiple upstream open reading frames and low mRNA abundance. These results suggest that the high levels of eIF4GI observed in many breast cancers might act to specifically increase proliferation, prevent autophagy, and release tumor cells from control by nutrient sensing. PMID- 18426978 TI - A cell-based reglucosylation assay demonstrates the role of GT1 in the quality control of a maturing glycoprotein. AB - The endoplasmic reticulum (ER) protein GT1 (UDP-glucose: glycoprotein glucosyltransferase) is the central enzyme that modifies N-linked carbohydrates based upon the properties of the polypeptide backbone of the maturing substrate. GT1 adds glucose residues to nonglucosylated proteins that fail the quality control test, supporting ER retention through persistent binding to the lectin chaperones calnexin and calreticulin. How GT1 functions in its native environment on a maturing substrate is poorly understood. We analyzed the reglucosylation of a maturing model glycoprotein, influenza hemagglutinin (HA), in the intact mammalian ER. GT1 reglucosylated N-linked glycans in the slow-folding stem domain of HA once the nascent chain was released from the ribosome. Maturation mutants that disrupted the oxidation or oligomerization of HA also supported region specific reglucosylation by GT1. Therefore, GT1 acts as an ER quality control sensor by posttranslationally reglucosylating glycans on slow-folding or nonnative domains to recruit chaperones specifically to critical aberrant regions. PMID- 18426979 TI - Remodeling of cellular cytoskeleton by the acid sphingomyelinase/ceramide pathway. AB - The chemotherapeutic agent cisplatin is widely used in treatment of solid tumors. In breast cancer cells, cisplatin produces early and marked changes in cell morphology and the actin cytoskeleton. These changes manifest as loss of lamellipodia/filopodia and appearance of membrane ruffles. Furthermore, cisplatin induces dephosphorylation of the actin-binding protein ezrin, and its relocation from membrane protrusions to the cytosol. Because cisplatin activates acid sphingomyelinase (ASMase), we investigate here the role of the ASMase/ceramide (Cer) pathway in mediating these morphological changes. We find that cisplatin induces a transient elevation in ASMase activity and its redistribution to the plasma membrane. This translocation is blocked upon overexpression of a dominant negative (DN) ASMase(S508A) mutant and by a DN PKCdelta. Importantly; knockdown of ASMase protects MCF-7 cells from cisplatin-induced cytoskeletal changes including ezrin dephosphorylation. Reciprocally, exogenous delivery of D-e-C16 Cer, but not dihydro-C16-Cer, recapitulates the morphotropic effects of cisplatin. Collectively, these results highlight a novel tumor suppressor property for Cer and a function for ASMase in cisplatin-induced cytoskeletal remodeling. PMID- 18426980 TI - ErbB2 directly activates the exchange factor Dock7 to promote Schwann cell migration. AB - The cellular events that precede myelination in the peripheral nervous system require rapid and dynamic morphological changes in the Schwann cell. These events are thought to be mainly controlled by axonal signals. But how signals on the axons are coordinately organized and transduced to promote proliferation, migration, radial sorting, and myelination is unknown. We describe that the axonal signal neuregulin-1 (NRG1) controls Schwann cell migration via activation of the atypical Dock180-related guanine nucleotide exchange factor (GEF) Dock7 and subsequent activation of the Rho guanine triphosphatases (GTPases) Rac1 and Cdc42 and the downstream c-Jun N-terminal kinase. We show that the NRG1 receptor ErbB2 directly binds and activates Dock7 by phosphorylating Tyr-1118. Dock7 knockdown, or expression of Dock7 harboring the Tyr-1118-to-Phe mutation in Schwann cells, attenuates the effects of NRG1. Thus, Dock7 functions as an intracellular substrate for ErbB2 to promote Schwann cell migration. This provides an unanticipated mechanism through which ligand-dependent tyrosine phosphorylation can trigger the activation of Rho GTPase-GEFs of the Dock180 family. PMID- 18426981 TI - A central function for perlecan in skeletal muscle and cardiovascular development. AB - Perlecan's developmental functions are difficult to dissect in placental animals because perlecan disruption is embryonic lethal. In contrast to mammals, cardiovascular function is not essential for early zebrafish development because the embryos obtain adequate oxygen by diffusion. In this study, we use targeted protein depletion coupled with protein-based rescue experiments to investigate the involvement of perlecan and its C-terminal domain V/endorepellin in zebrafish development. The perlecan morphants show a severe myopathy characterized by abnormal actin filament orientation and disorganized sarcomeres, suggesting an involvement of perlecan in myopathies. In the perlecan morphants, primary intersegmental vessel sprouts, which develop through angiogenesis, fail to extend and show reduced protrusive activity. Live videomicroscopy confirms the abnormal swimming pattern caused by the myopathy and anomalous head and trunk vessel circulation. The phenotype is partially rescued by microinjection of human perlecan or endorepellin. These findings indicate that perlecan is essential for the integrity of somitic muscle and developmental angiogenesis and that endorepellin mediates most of these biological activities. PMID- 18426983 TI - IL-6-dependent spontaneous proliferation is required for the induction of colitogenic IL-17-producing CD8+ T cells. AB - We propose a novel role for interleukin (IL) 6 in inducing rapid spontaneous proliferation (SP) of naive CD8(+) T cells, which is a crucial step in the differentiation of colitogenic CD8(+) T cells. Homeostasis of T cells is regulated by two distinct modes of cell proliferation: major histocompatibility complex/antigen-driven rapid SP and IL-7/IL-15-dependent slow homeostatic proliferation. Using our novel model of CD8(+) T cell-dependent colitis, we found that SP of naive CD8(+) T cells is essential for inducing pathogenic cytokine producing effector T cells. The rapid SP was predominantly induced in mesenteric lymph nodes (LNs) but not in peripheral LNs under the influence of intestinal flora and IL-6. Indeed, this SP was markedly inhibited by treatment with anti-IL 6 receptor monoclonal antibody (IL-6R mAb) or antibiotic-induced flora depletion, but not by anti-IL-7R mAb and/or in IL-15-deficient conditions. Concomitantly with the inhibition of SP, anti-IL-6R mAb significantly inhibited the induction of CD8(+) T cell-dependent autoimmune colitis. Notably, the transfer of naive CD8(+) T cells derived from IL-17(-/-) mice did not induce autoimmune colitis. Thus, we conclude that IL-6 signaling is crucial for SP under lymphopenic conditions, which subsequently caused severe IL-17-producing CD8(+) T cell mediated autoimmune colitis. We suggest that anti-IL-6R mAb may become a promising strategy for the therapy of colitis. PMID- 18426984 TI - Virulent Shigella flexneri subverts the host innate immune response through manipulation of antimicrobial peptide gene expression. AB - Antimicrobial factors are efficient defense components of the innate immunity, playing a crucial role in the intestinal homeostasis and protection against pathogens. In this study, we report that upon infection of polarized human intestinal cells in vitro, virulent Shigella flexneri suppress transcription of several genes encoding antimicrobial cationic peptides, particularly the human beta-defensin hBD-3, which we show to be especially active against S. flexneri. This is an example of targeted survival strategy. We also identify the MxiE bacterial regulator, which controls a regulon encompassing a set of virulence plasmid-encoded effectors injected into host cells and regulating innate signaling, as being responsible for this dedicated regulatory process. In vivo, in a model of human intestinal xenotransplant, we confirm at the transcriptional and translational level, the presence of a dedicated MxiE-dependent system allowing S. flexneri to suppress expression of antimicrobial cationic peptides and promoting its deeper progression toward intestinal crypts. We demonstrate that this system is also able to down-regulate additional innate immunity genes, such as the chemokine CCL20 gene, leading to compromised recruitment of dendritic cells to the lamina propria of infected tissues. Thus, S. flexneri has developed a dedicated strategy to weaken the innate immunity to manage its survival and colonization ability in the intestine. PMID- 18426985 TI - Essential role of Notch signaling in effector memory CD8+ T cell-mediated airway hyperresponsiveness and inflammation. AB - Adoptive transfer of in vivo-primed CD8(+) T cells or in vitro-generated effector memory CD8(+) T (T(EFF)) cells restores airway hyperresponsiveness (AHR) and airway inflammation in CD8-deficient (CD8(-/-)) mice. Examining transcription levels, there was a strong induction of Notch1 in T(EFF) cells compared with central memory CD8(+) T cells. Treatment of T(EFF) cells with a gamma-secretase inhibitor (GSI) strongly inhibited Notch signaling in these cells, and after adoptive transfer, GSI-treated T(EFF) cells failed to restore AHR and airway inflammation in sensitized and challenged recipient CD8(-/-) mice, or to enhance these responses in recipient wild-type (WT) mice. These effects of GSI were also associated with increased expression of the Notch ligand Delta1 in T(EFF) cells. Treatment of sensitized and challenged WT mice with Delta1-Fc resulted in decreased AHR and airway inflammation accompanied by higher levels of interferon gamma in bronchoalveolar lavage fluid. These results demonstrate a role for Notch in skewing the T cell response from a T helper (Th)2 to a Th1 phenotype as a consequence of the inhibition of Notch receptor activation and the up-regulation of the Notch ligand Delta1. These data are the first to show a functional role for Notch in the challenge phase of CD8(+) T cell-mediated development of AHR and airway inflammation, and identify Delta1 as an important regulator of allergic airway inflammation. PMID- 18426986 TI - C5 deficiency and C5a or C5aR blockade protects against cerebral malaria. AB - Experimental infection of mice with Plasmodium berghei ANKA (PbA) provides a powerful model to define genetic determinants that regulate the development of cerebral malaria (CM). Based on the hypothesis that excessive activation of the complement system may confer susceptibility to CM, we investigated the role of C5/C5a in the development of CM. We show a spectrum of susceptibility to PbA in a panel of inbred mice; all CM-susceptible mice examined were found to be C5 sufficient, whereas all C5-deficient strains were resistant to CM. Transfer of the C5-defective allele from an A/J (CM resistant) onto a C57BL/6 (CM susceptible) genetic background in a congenic strain conferred increased resistance to CM; conversely, transfer of the C5-sufficient allele from the C57BL/6 onto the A/J background recapitulated the CM-susceptible phenotype. The role of C5 was further explored in B10.D2 mice, which are identical for all loci other than C5. C5-deficient B10.D2 mice were protected from CM, whereas C5 sufficient B10.D2 mice were susceptible. Antibody blockade of C5a or C5a receptor (C5aR) rescued susceptible mice from CM. In vitro studies showed that C5a potentiated cytokine secretion induced by the malaria product P. falciparum glycosylphosphatidylinositol and C5aR blockade abrogated these amplified responses. These data provide evidence implicating C5/C5a in the pathogenesis of CM. PMID- 18426987 TI - Transmission of HIV-1 Gag immune escape mutations is associated with reduced viral load in linked recipients. AB - In a study of 114 epidemiologically linked Zambian transmission pairs, we evaluated the impact of human leukocyte antigen class I (HLA-I)-associated amino acid polymorphisms, presumed to reflect cytotoxic T lymphocyte (CTL) escape in Gag and Nef of the virus transmitted from the chronically infected donor, on the plasma viral load (VL) in matched recipients 6 mo after infection. CTL escape mutations in Gag and Nef were seen in the donors, which were subsequently transmitted to recipients, largely unchanged soon after infection. We observed a significant correlation between the number of Gag escape mutations targeted by specific HLA-B allele-restricted CTLs and reduced VLs in the recipients. This negative correlation was most evident in newly infected individuals, whose HLA alleles were unable to effectively target Gag and select for CTL escape mutations in this gene. Nef mutations in the donor had no impact on VL in the recipient. Thus, broad Gag-specific CTL responses capable of driving virus escape in the donor may be of clinical benefit to both the donor and recipient. In addition to their direct implications for HIV-1 vaccine design, these data suggest that CTL induced viral polymorphisms and their associated in vivo viral fitness costs could have a significant impact on HIV-1 pathogenesis. PMID- 18426988 TI - Rapid mobilization of functional donor hematopoietic cells without G-CSF using AMD3100, an antagonist of the CXCR4/SDF-1 interaction. AB - Allografts from HLA-matched sibling donors were mobilized and collected without granulocyte colony-stimulating factor (G-CSF) using AMD3100, a direct antagonist of CXCR4/stromal-derived factor 1 (SDF-1/CXCL12). Donors (N = 25) were treated with AMD3100 at a dose of 240 mug/kg by subcutaneous injection, and leukapheresis was then initiated just 4 hours later. Two-thirds of the donors collected an allograft with a CD34(+) cell dose sufficient for transplantation after just one dose of AMD3100. No donor experienced more than grade 1 toxicity. After a myeloablative regimen, 20 patients with hematologic malignancies received allografts collected after AMD3100 alone. All patients engrafted neutrophils (median day 10) and platelets (median day 12) promptly. Acute graft-versus-host disease (GVHD) grades 2 through 4 occurred in 35% of patients. One patient died due to complications related to acute GVHD. No unexpected adverse events were observed in any of the recipients. All 14 patients surviving in remission have robust trilineage hematopoiesis and are transfusion-free with a median follow-up of 277 days (range, 139-964 days). Direct antagonism of CXCR4 by AMD3100 may provide a more rapid and possibly less toxic and cumbersome alternative to traditional G-CSF-based mobilization in normal donors. This trial was registered as no. NCT00241358 at www.ClinicalTrials.gov. PMID- 18426990 TI - Genetic variants of inflammatory markers and arterial stiffness. PMID- 18426989 TI - MDM2 SNP309 and TP53 Arg72Pro interact to alter therapy-related acute myeloid leukemia susceptibility. AB - The p53 tumor suppressor directs the cellular response to many mechanistically distinct DNA-damaging agents and is selected against during the pathogenesis of therapy-related acute myeloid leukemia (t-AML). We hypothesized that constitutional genetic variation in the p53 pathway would affect t-AML risk. Therefore, we tested associations between patients with t-AML (n = 171) and 2 common functional p53-pathway variants, the MDM2 SNP309 and the TP53 codon 72 polymorphism. Although neither polymorphism alone influenced the risk of t-AML, an interactive effect was detected such that MDM2 TT TP53 Arg/Arg double homozygotes, and individuals carrying both a MDM2 G allele and a TP53 Pro allele, were at increased risk of t-AML (P value for interaction is .009). This interactive effect was observed in patients previously treated with chemotherapy but not in patients treated with radiotherapy, and in patients with loss of chromosomes 5 and/or 7, acquired abnormalities associated with prior exposure to alkylator chemotherapy. In addition, there was a trend toward shorter latency to t-AML in MDM2 GG versus TT homozygotes in females but not in males, and in younger but not older patients. These data indicate that the MDM2 and TP53 variants interact to modulate responses to genotoxic therapy and are determinants of risk for t-AML. PMID- 18426991 TI - Valsartan improves arterial stiffness in type 2 diabetes independently of blood pressure lowering. AB - Increased arterial stiffness, as estimated from aortic pulse wave velocity (Ao PWV), and albuminuria are independent predictors for cardiovascular disease in type 2 diabetes mellitus (T2DM). Whether angiotensin receptor blockers (ARBs), drugs with cardio-renal protective effects, improve Ao-PWV to a greater extent than other equipotent antihypertensive medications remains unclear. After a 4 week washout phase, we compared the effects of valsartan (n=66), an ARB, with that of amlodipine (n=65), a calcium channel blocker on Ao-PWV in 131 T2DM patients with pulse pressure (PP) >or=60 mm Hg and raised albumin excretion rate (AER) in a 24-week randomized, double-blind, parallel group study. Hydrochlorothiazide (HCTZ) 25 mg/d was added to valsartan 160 mg and amlodipine 5 mg/od uptitrated to 10 mg/od after 4 weeks to ensure equivalent BP control. After 24 weeks brachial and central aortic PP had fallen to a similar extent with attained mean (SD) brachial and central PP of 61.6 (13.6) and 47.3 (14.1) mm Hg in the valsartan/HCTZ group and 61.5 (12.2) and 47.3 (9.9) mm Hg in the amlodipine group, respectively. Ao-PWV showed a significantly greater reduction, mean (95% CI), -0.9 m/s (-1.4 to -0.3) for valsartan/HCTZ compared to amlodipine (P=0.002). AER fell significantly only with Val/HCTZ from 30.8(20.4, 46.5) to 18.2(12.5, 26.3) mcg/min, (P=0.01) with between treatment difference in favor of Val/HCTZ of -15.3mcg/min (P<0.001). Changes in AER and Ao-PWV were not correlated. Valsartan/HCTZ improves arterial stiffness and AER to a significantly greater extent than amlodipine despite similar central and brachial BP control. These 2 effects, which appear independent of each other, may explain the specific cardio-renal protective properties of ARBs. PMID- 18426992 TI - Collecting duct renin is upregulated in both kidneys of 2-kidney, 1-clip goldblatt hypertensive rats. AB - Renin in collecting duct cells is upregulated in chronic angiotensin II-infused rats via angiotensin II type 1 receptors. To determine whether stimulation of collecting duct renin is a blood pressure-dependent effect; changes in collecting duct renin and associated parameters were assessed in both kidneys of 2-kidney, 1 clip Goldblatt hypertensive (2K1C) rats. Renal medullary tissues were used to avoid the contribution of renin from juxtaglomerular cells. Systolic blood pressure increased to 184+/-9 mm Hg in 2K1C rats (n=19) compared with sham rats (121+/-6 mm Hg; n=12). Although renin immunoreactivity markedly decreased in juxtaglomerular cells of nonclipped kidneys (NCK: 0.2+/-0.0 versus 1.0+/-0.0 relative ratio) and was augmented in clipped kidneys (CK: 1.7+/-1.0 versus 1.0+/ 0.0 relative ratio), its immunoreactivity increased in cortical and medullary collecting ducts of both kidneys of 2K1C rats (CK: 2.8+/-1.0 cortex; 2.1+/-1.0 medulla; NCK: 4.6+/-2.0 cortex, 3.2+/-1.0 medulla versus 1.0+/-0.0 in sham kidneys). Renal medullary tissues of 2K1C rats showed greater levels of renin protein (CK: 1.4+/-0.2; NCK: 1.5+/-0.3), renin mRNA (CK: 5.8+/-2.0; NCK: 4.9+/ 2.0), angiotensin I (CK: 120+/-18 pg/g; NCK: 129+/-13 pg/g versus sham: 67+/-6 pg/g), angiotensin II (CK: 150+/-32 pg/g; NCK: 123+/-21 pg/g versus sham: 91+/-12 pg/g; P<0.05), and renin activity (CK: 8.6 microg of angiotensin I per microgram of protein; NCK: 8.3 microg of angiotensin I per microgram of protein; sham: 3.4 microg of angiotensin I per microgram of protein) than sham rats. These data indicate that enhanced collecting duct renin in 2K1C rats occurs independently of blood pressure. Upregulation of distal tubular renin helps to explain how sustained intrarenal angiotensin II formation occurs even during juxtaglomerular renin suppression, thus allowing maintained effects on tubular sodium reabsorption that contribute to the hypertension. PMID- 18426993 TI - Hypertension impairs postnatal vasculogenesis: role of antihypertensive agents. AB - We analyzed the effect of hypertension on postischemic vasculogenesis. Ischemia was induced by right femoral artery ligature in Wistar Kyoto rats (WKY) or spontaneously hypertensive rats (SHR) treated with or without angiotensin converting enzyme inhibitor (Perindopril, 0.76 mg/kg/d) and angiotensin type 1 receptor blocker (losartan, 30 mg/kg/d). Basal postischemic neovascularization was reduced in SHR compared to WKY (P<0.05, n=8). Treatment with ACE inhibitor or angiotensin type 1 receptor blocker decreased blood pressure levels by 1.4- and 1.3-fold (P<0.001), respectively and restored vessel growth in SHR to WKY levels. Interestingly, 14 days after bone-marrow mononuclear cell (BM-MNC) transfusion, angiographic scores, capillary density, and foot perfusion were decreased by 1.4 , 1.5-, and 1.2-fold, respectively in SHR transfused with BM-MNCs isolated from SHR compared to those receiving BM-MNCs of WKY (P<0.05, n=6). Alteration in BM MNCs proangiogenic potential was likely related to the reduction in their ability to mobilize into peripheral circulation, as revealed by the 2.9-fold decrease in number of circulating CD34+/CD117+ cells (P<0.001) and to differentiate into cells with endothelial phenotype, as revealed by the 2.1-fold reduction in percentages of DilLDL/BS-1 lectin positive cells (P<0.001). In addition, reactive oxygen species (ROS) levels were increased by 2.2-fold in SHR BM-MNCs compared to WKY BM-MNCs (P<0.01), as assessed by L-012 luminescence. Cotreatment with ACE inhibitor, angiotensin type 1 receptor blocker, or antioxidants (NAC 3 mmol/L, Apocynin 200 micromol/L) reduced ROS levels, improved the number of DilLDL/BS-1 lectin-positive cells by around 1.5-fold, and restored BM-MNCs proangiogenic effects in ischemic hindlimb. In conclusion, alteration in progenitor cell proangiogenic function may participate to the hypertension-induced impairment in postischemic revascularization. PMID- 18426994 TI - Central aortic blood pressure and cardiovascular risk: a paradigm shift? PMID- 18426995 TI - Forearm vascular reactivity and arterial stiffness in asymptomatic adults from the community. AB - Vascular reactivity may affect the stiffness characteristics of the arterial wall. We investigated the association between forearm microcirculatory and conduit artery function and measures of arterial stiffness in 527 asymptomatic non-Hispanic white adults without known cardiovascular disease. High-resolution ultrasonography of the brachial artery (ba) was performed to assess forearm microcirculatory function (ba blood flow velocity, local shear stress, and forearm vascular resistance at rest and during reactive hyperemia) and conduit artery function (ba flow-mediated dilatation [baFMD] and ba nitroglycerin mediated dilatation [baNMD]). Arterial stiffness was assessed by cuff-derived brachial pulse pressure and aortic pulse wave velocity (aPWV) measured by applanation tonometry. In regression analyses that adjusted for heart rate, mean arterial pressure, height, cardiovascular risk factors, and hypertension medication and statin use, higher baseline ba systolic velocity and systolic shear stress were associated with greater pulse pressure (P=0.0002 and P=0.006, respectively) and higher aPWV (each P<0.0001). During hyperemia, lower ba mean velocity and lower mean shear stress were associated with higher pulse pressure (P=0.045 and P=0.036, respectively), whereas both systolic and mean velocity (P<0.0001 and P=0.002, respectively) and systolic and mean shear stress (P<0.0001 and P=0.003, respectively) were inversely associated with aPWV. baFMD was not associated with pulse pressure but was inversely associated with aPWV (P=0.011). baNMD was inversely associated with pulse pressure (P=0.0002) and aPWV (P=0.008). Our findings demonstrate that impaired forearm microvascular function (in the form of elevated resting blood flow velocity and impaired flow reserve) and impaired brachial artery reactivity are associated with increased arterial stiffness. PMID- 18426996 TI - Relations of inflammatory biomarkers and common genetic variants with arterial stiffness and wave reflection. AB - Inflammation causes vascular dysfunction and perpetuates proatherosclerotic processes. We hypothesized that a broad panel of inflammatory biomarkers and single nucleotide polymorphisms in inflammatory genes is associated with vascular stiffness. We assessed 12 circulating inflammatory biomarkers (C-reactive protein, fibrinogen, interleukin-6, intercellular adhesion molecule-1, lipoprotein-associated phospholipase-A2 [mass and activity], monocyte chemoattractant protein-1, myeloperoxidase, CD40 ligand, osteoprotegerin, P selectin, and tumor necrosis factor receptor-II) in relation to tonometry variables (central pulse pressure, mean arterial pressure, forward pressure wave, reflected pressure wave, carotid-femoral pulse wave velocity, and augmentation index) measured in 2409 Framingham Heart Study participants (mean age: 60 years; 55% women; 13% ethnic/racial minorities). Single nucleotide polymorphisms (n=2195) in 240 inflammatory candidate genes were related to tonometry measures in 1036 white individuals. In multivariable analyses, biomarkers explained <1% of any tonometry measure variance. Applying backward elimination, markers related to tonometry (P<0.01) were as follows: tumor necrosis factor receptor-II (inversely) with mean arterial pressure; C-reactive protein (positively) and lipoprotein associated phospholipase-A2 (inversely) with reflected pressure wave; and interleukin-6 and osteoprotegerin (positively) with carotid-femoral pulse wave velocity. In genetic association analyses, lowest P values (false discovery rate <0.50) were observed for rs10509561 (FAS), P=6.6x10(-5) for central pulse pressure and rs11559271 (ITGB2), P=1.1x10(-4) for mean arterial pressure. These data demonstrate that, in a community-based sample, circulating inflammatory markers tumor necrosis factor receptor-II (mean arterial pressure), C-reactive protein, lipoprotein-associated phospholipase-A2 activity (reflected pressure wave), interleukin-6, and osteoprotegerin (carotid-femoral pulse wave velocity) were significantly but modestly associated with measures of arterial stiffness and wave reflection. Additional studies are needed to determine whether variation in inflammatory marker genes is associated with tonometry measures. PMID- 18426997 TI - Central pressure: variability and impact of cardiovascular risk factors: the Anglo-Cardiff Collaborative Trial II. AB - Pulse pressure varies throughout the arterial tree, resulting in a gradient between central and peripheral pressure. Factors such as age, heart rate, and height influence this gradient. However, the relative impact of cardiovascular risk factors and atheromatous disease on central pressure and the normal variation in central pressure in healthy individuals are unclear. Seated peripheral (brachial) and central (aortic) blood pressures were assessed, and the ratio between aortic and brachial pulse pressure (pulse pressure ratio, ie, 1/amplification) was calculated in healthy individuals, diabetic subjects, patients with cardiovascular disease, and in individuals with only 1 of the following: hypertension, hypercholesterolemia, or smoking. The age range was 18 to 101 years, and data from 10 613 individuals were analyzed. Compared with healthy individuals, pulse pressure ratio was significantly increased (ie, central systolic pressure was relatively higher) in individuals with risk factors or disease (P<0.01 for all of the comparisons). Although aging was associated with an increased pulse pressure ratio, there was still an average+/-SD difference between brachial and aortic systolic pressure of 11+/-4 and 8+/-3 mm Hg for men and women aged >80 years, respectively. Finally, stratifying individuals by brachial pressure revealed considerable overlap in aortic pressure, such that >70% of individuals with high-normal brachial pressure had similar aortic pressures as those with stage 1 hypertension. These data demonstrate that cardiovascular risk factors affect the pulse pressure ratio, and that central pressure cannot be reliably inferred from peripheral pressure. However, assessment of central pressure may improve the identification and management of patients with elevated cardiovascular risk. PMID- 18426998 TI - Excessive nitric oxide function and blood pressure regulation in patients with autonomic failure. AB - Approximately 50% of patients with autonomic failure (AF) suffer from supine hypertension, even those with very low plasma norepinephrine and renin. Because NO is arguably the most potent metabolic modulator of blood pressure, we hypothesized that impaired NO function contributes to supine hypertension in AF. However, we found that AF patients (n=14) were more sensitive to the pressor effects of the NO synthase inhibitor N(G)-monomethyl-l-arginine, suggesting increased NO function rather than deficiency; a lower dose of N(G)-monomethyl-l arginine was needed to produce a similar increase in blood pressure in AF patients, as in healthy control subjects in whom AF was induced with the ganglionic blocker trimethaphan (171+/-37 mg versus 512+/-81 mg, respectively; P=0.001). Furthermore, potentiation of the actions of endogenous NO with the phosphodiesterase inhibitor sildenafil (25 mg PO) decreased nighttime supine systolic blood pressure from 182+/-11 to 138+/-4 mm Hg in 8 AF patients with supine hypertension (P=0.012 compared with placebo). Finally, AF patients tolerated a greater degree of upright tilt during infusion of N(G)-monomethyl-l arginine (56+/-6 degrees versus 41+/-4 degrees with placebo; n=7; P=0.014), an improvement in orthostatic tolerance similar to that obtained with equipressor doses of phenylephrine. In conclusion, AF patients do not have NO deficiency contributing to supine hypertension. Instead, they have increased NO function contributing to their orthostatic hypotension. Potentiation of NO could be used in the treatment of supine hypertension, and its inhibition offers a novel approach to improve orthostatic hypotension. PMID- 18426999 TI - Myocardial protection against pressure overload in mice lacking Bach1, a transcriptional repressor of heme oxygenase-1. AB - Bach1 is a stress-responsive transcriptional factor that is thought to control the expression levels of cytoprotective factors, including heme-oxygenase (HO)-1. In the present study, we investigated the roles of Bach1 in the development of left ventricular (LV) hypertrophy and remodeling induced by transverse aortic constriction (TAC) in vivo using Bach1 gene-deficient (Bach1(-/-)) mice. TAC for 3 weeks in wild-type control (Bach1(+/+)) mice produced LV hypertrophy and remodeling manifested by increased heart weight, histological findings showing increased myocyte cross-sectional area (CSA) and interstitial fibrosis (picro Sirius red staining), reexpressions of ANP, BNP, and betaMHC genes, and echocardiographic findings showing wall thickening, LV dilatation, and reduced LV contraction. Deletion of Bach1 caused significant reductions in heart weight (by 16%), CSA (by 36%), tissue collagen content (by 38%), and gene expression levels of ANP (by 75%), BNP (by 45%), and betaMHC (by 74%). Echocardiography revealed reduced LV dimension and ameliorated LV contractile function. Deletion of Bach1 in the LV caused marked upregulation of HO-1 protein accompanied by elevated HO activity in both basal or TAC-stimulated conditions. Treatment of Bach1(-/-) mice with tin-protoporphyrin, an inhibitor of HO, abolished the antihypertrophic and antiremodeling effects of Bach1 gene ablation. These results suggest that deletion of Bach1 caused upregulation of cytoprotective HO-1, thereby inhibiting TAC-induced LV hypertrophy and remodeling, at least in part, through activation of HO. Bach1 repressively controls myocardial HO-1 expression both in basal and stressed conditions, inhibition of Bach1 may be a novel therapeutic strategy to protect the myocardium from pressure overload. PMID- 18427000 TI - Modulation of cellular invasion by VEGF-C expression in squamous cell carcinoma of the head and neck. AB - OBJECTIVE: To determine how vascular endothelial growth factor C (VEGF-C) affects tumor cell invasion and motility in squamous cell carcinoma of the head and neck (SCCHN). DESIGN: A molecular biology study. The VEGF-C coding sequence was cloned into an expression vector and stably transfected into the SCCHN cell line SCC116 to create the SCC116-VEGFC line. RNA interference (RNAi) was used to block VEGF-C expression. An adenoviral system for expressing VEGF-C RNAi was developed and tested. SETTING: An academic hospital laboratory. MAIN OUTCOME MEASURES: Relative VEGF-C RNA levels were determined by real-time quantitative reverse transcriptase polymerase chain reaction, and protein expression was evaluated by Western blot. Cellular invasion was evaluated by 24-hour semipermeable membrane transit assay. RESULTS: SCC116-VEGFC cells had markedly increased expression of VEGF-C protein and RNA compared with normal SCC116 controls. SCC116-VEGFC cells produced marked increases in cellular invasion and motility compared with SCC116 cells. Blockade of VEGF-C expression by transfection of a VEGF-C RNAi expression plasmid into both SCC116 and SCC116-VEGFC cells induced a 38% decrease in SCCHN invasion and motility as tested by a semipermeable membrane invasion assay. We developed an adenoviral expression system for VEGF-C RNAi, which also induced a dose-dependent decrease in cellular invasion in the highly invasive DM12 cell line. CONCLUSIONS: These studies demonstrate that intracellular VEGF-C levels modulate in vitro SCCHN motility and invasion. Further work is needed to clarify the specific receptors and signaling pathways that are involved in SCCHN motility. Molecular therapies that inhibit the VEGF-C pathway may have clinical potential in the treatment of lymphatic metastasis in SCCHN. PMID- 18427001 TI - Expression of p53 and Bcl-xL as predictive markers for larynx preservation in advanced laryngeal cancer. AB - OBJECTIVE: To assess tumor markers in advanced laryngeal cancer. DESIGN: Marker expression and clinical outcome. PATIENTS: Pretreatment tumor biopsy specimens were analyzed from patients enrolled in the Department of Veterans Affairs Laryngeal Cancer Study. MAIN OUTCOME MEASURES: Expression of p53 (OMIM TP53) and Bcl-xL (OMIM 600039) in pretreatment biopsy specimens was assessed for correlation with chemotherapy response, laryngeal preservation, and survival. RESULTS: Higher rates of larynx preservation were observed in patients whose tumors expressed p53 vs those that did not (80% [36 of 45 patients] vs 59% [24 of 41 patients], P =.03). Higher rates of larynx preservation were also observed in patients whose tumors expressed low levels of Bcl-xL vs high levels of Bcl-xL (90% [18 of 20 patients] vs 60% [30 of 50 patients], P =.02). Patients were categorized into 3 risk groups (low, intermediate, and high) based on their tumor p53 and Bcl-xL expression status. Patients whose tumors had the high-risk biomarker profile (low p53 expression and high Bcl-xL expression) were less likely to preserve their larynx than patients whose tumors had the intermediate risk biomarker profile (high p53 expression and low or high Bcl-xL expression) or the low-risk biomarker profile (low p53 expression and low Bcl-xL expression). The larynx preservation rates were 100% (10 of 10 patients), 77% (26 of 34 patients), and 54% (7 of 13 patients) for the low-risk, intermediate-risk, and high-risk groups, respectively (P =.04, Fisher exact test). CONCLUSION: Tumor expression of p53 and Bcl-xL is a strong predictor of successful larynx preservation in patients treated with induction chemotherapy and followed by radiation therapy in responding tumors. PMID- 18427002 TI - Five-year survival rates and time trends of laryngeal cancer in the US population. AB - OBJECTIVES: To provide comprehensive temporal trend analysis of 5-year relative survival rates of laryngeal cancer using the Surveillance, Epidemiology, and End Results database; and to expand on prior reports by including inclusion of laryngeal tumor location, stage, age at diagnosis, treatment strategy, and histologic grade. DESIGN: Retrospective cohort analysis using the Surveillance, Epidemiology, and End Results database of the National Cancer Institute. The Surveillance, Epidemiology, and End Results data were used to design 5 cohorts of patients with laryngeal cancer: 1977-1978, 1983-1984, 1989-1990, 1995-1996, and 2001-2002. Five-year survival rates were analyzed according to tumor site, stage, and grade; age at diagnosis; and treatment strategy. The joinpoint regression model was used to assess survival trends and their statistical significance. RESULTS: Among patients with supraglottic cancer, 5-year relative survival rates for distant disease worsened over time while rates for local and regional disease did not change (P = .01 and P > .05, respectively). For localized glottic cancer, survival remained stable from 1977-1978 to 2001-2002. However, patients with regional and distant glottic cancer demonstrated a significant decrease in survival in the past 3 decades (P < .001). This trend was independent of treatment strategy. Finally, the proportion of well-differentiated tumors in patients with regional laryngeal cancer decreased over time (P < .001 for supraglottic and P = .007 for glottic). CONCLUSIONS: A decreasing 5-year survival trend was found among patients with glottic cancer who had regional disease and in all patients with distant disease. Histopathologic trends not previously reported in those with laryngeal cancer seem to parallel those seen in other tobacco-related cancers. These trends may reflect the effect of birth cohorts and implicate the relationship between carcinogenic exposure and host factors, rather than the influence of treatment. PMID- 18427003 TI - Prognostic indicators of unilateral vocal fold paralysis. AB - OBJECTIVES: To determine the possible prognostic indicators of unilateral vocal fold paralysis (UVFP) and survey the timing and values of preset laryngeal electromyography (LEMG) rules for UVFP. DESIGN: Cohort study with retrospective data analysis. SETTING: Voice clinic of a tertiary medical center. PATIENTS: Complete data for 45 patients diagnosed with idiopathic or iatrogenic UVFP. The LEMG was performed between 3 weeks and 6 months from the onset of symptoms. MAIN OUTCOME MEASURE: At least 6 months after symptom onset and 3 months after LEMG. RESULTS: Thirteen subjects showed resolved vocal fold motion (29%), and 32 had persistent vocal fold paralysis (71%). According to the LEMG decision rules proposed by Munin et al in 2003, the predictive values for positive results, negative results, sensitivity, specificity, and accuracy of LEMG were 78.9%, 71.4%, 93.8%, 38.5%, and 77.8%, respectively. We found the false-positive rate to be as high as 50% if LEMG was performed less than 2 months after symptom onset, and only 7.7% if LEMG was performed at least 2 months after symptom onset. After excluding 14 LEMG data recorded at less than 2 months, the predictive values for positive results, negative results, sensitivity, specificity, and accuracy of LEMG were 92.3%, 60%, 92.3%, 60.0%, and 87.1%, respectively. The predictive values of positive results and accuracy significantly improved without compromising sensitivity. CONCLUSION: This study confirms that LEMG is a clinically useful tool that can offer prognostic information for UVFP especially if it is done at least 2 months after symptom onset. PMID- 18427004 TI - Temporomandibular disorder and new aural symptoms. AB - OBJECTIVES: To report the prevalence and demographics of temporomandibular disorder (TMD) within a population of clinic patients and to describe the prevalence of revisited and new, previously unstudied, aural symptoms described by a sample of these patients with TMD (hereinafter "TMD patients"). DESIGN: A retrospective evaluation of patient records was completed to determine the percentage and the demographics of TMD patients in a clinical setting. A prospective analysis was done on the self-reported prevalence of previously studied and new aural symptoms of 78 TMD study patients compared with 78 control patients without TMD. SETTING: A private otolaryngology practice in a rural Arizona town. PATIENTS: Patients with TMD and aural symptoms. RESULTS: Ten percent of all new otolaryngology clinic patients were diagnosed as having TMD. Of the 78 patients, 27 (35%) listed the ear as one of their sites of pain. The prevalence of each of the 8 aural symptoms assessed was significantly higher in TMD patients compared with controls (P < .001). A warm and/or fluid sensation in the ear and a stuffed cotton sensation in the ear were the most indicative symptoms of TMD because they had the highest relative risk ratios in TMD patients. Aural symptoms of loud noise sensitivity and cold air/wind sensitivity are also relevant and were approximately 5 times more frequent in TMD subjects than in controls. CONCLUSIONS: Patients with TMD are a significant component of otolaryngology practice. There are previously uninvestigated aural symptoms that occur much more frequently in TMD patients than in patients without TMD. PMID- 18427005 TI - Subjective visual vertical and horizontal: effect of the preset angle. AB - OBJECTIVES: (1) To study the subjective visual vertical (SVV) and subjective visual horizontal (SVH) in patients with long-standing unilateral peripheral vestibular dysfunction (PVD) and unilateral Meniere's disease (MD) compared with controls. (2) To study the relationship between the direction of deviation of the linear marker (preset angle) and measures of SVV and SVH values. DESIGN: Prospective case-control study. SETTING: Outpatient clinic in a tertiary neuro otology department. PATIENTS: Seventeen healthy volunteers (mean age, 35.5 years), 9 patients with PVD (mean age, 43.1 years), and 10 patients with MD (mean age, 50.7 years) were included in the analysis. INTERVENTIONS: All subjects had a detailed neuro-otological evaluation. Twelve replicate readings of SVV and SVH were taken for each subject, with random preset angles, 6 in the clockwise and 6 in the counterclockwise direction. MAIN OUTCOME MEASURE: The SVV and SVH values were correlated with clinical features and the direction of the preset angle. RESULTS: The 2 subjects with PVD who had abnormal mean SVV and SVH values had symptoms of dysequilibrium and otolithic involvement. The 5 patients in the MD group who had abnormal mean SVV and SVH values had either recent acute vertiginous attacks or total canal paresis on the affected side. A previously unreported finding, to our knowledge, is that the SVV value depends on the direction of the preset angle in all subject groups, more so in the PVD and MD groups compared with controls. The SVV is inclined toward the direction of the preset angle. A weaker relation is seen between the SVH and preset angle. CONCLUSION: The preset angle should be considered when comparing SVV and SVH values. PMID- 18427006 TI - A novel SLC26A4 (PDS) deafness mutation retained in the endoplasmic reticulum. AB - OBJECTIVES: To identify mutations in the SLC26A4 gene in individuals with nonsyndromic hearing loss and enlarged vestibular aqueduct, to design a predicted model of the pendrin protein, and to characterize novel mutations by means of localization in mammalian cells and effect of the mutation on the predicted model. DESIGN: Validation of the mutation by its exclusion in more than 300 individuals with normal hearing. SETTING: A laboratory of genetics of hearing loss research, clinical genetics laboratories, an otolaryngology department at Tel Aviv University, and medical centers in Israel. PATIENTS: A patient with nonsyndromic hearing loss and enlarged vestibular aqueduct, 203 deaf probands, and 310 controls with normal hearing. INTERVENTIONS: Sequencing the SLC26A4 gene in the patient with nonsyndromic hearing loss and enlarged vestibular aqueduct. Transfection of yellow fluorescent protein (YFP) constructs into mammalian COS7 cells. Designing a computational model of the human SLC26A4 protein. MAIN OUTCOME MEASURE: Detection of a novel c.1458_1459insT SLC26A4 mutation. RESULTS: A computational model of the human pendrin protein suggests that the novel c.1458_1459insT mutation leads to a prematurely truncated protein, p.Ile487TyrfsX39. Mammalian COS7 cells transfected with the YFP-1458_1459insT construct showed mislocalization of the mutant protein. CONCLUSIONS: A novel SLC26A4 mutation was detected in Israel. Because current estimates demonstrate that SLC26A4 mutations are involved in up to 4% of nonsyndromic deafness, our findings emphasize the importance of adding a molecular test for the SLC26A4 gene in the diagnosis of deafness, particularly when bone abnormalities are involved, to the list of genes screened in Israel and elsewhere in the world. PMID- 18427007 TI - Pediatric mediastinitis as a complication of methicillin-resistant Staphylococcus aureus retropharyngeal abscess. AB - OBJECTIVE: To examine changes in the incidence, bacteriology, and complications of retropharyngeal infection (RPI) over an 8-year period. DESIGN: Retrospective medical record review. SETTING: Tertiary children's hospital. PATIENTS: The study population comprised 108 patients younger than 18 years old. INTERVENTION: Medical record review of patients with a discharge diagnosis of RPI (International Classification of Diseases, Ninth Revision code 478.24). MAIN OUTCOME MEASURES: Cases from June 1997 to May 2001 were compared with those from June 2001 to May 2005 to examine changes in the incidence, bacteriology, and complications of RPI. RESULTS: The number of RPI cases doubled from 36 to 72 in the final 4 years. In the first 4 years, no isolates of methicillin-resistant Staphylococcus aureus (MRSA) were found, and 1 patient developed mediastinitis. In the last 4 years, 8 of 25 patients (32%) with positive cultures had MRSA isolated, and 7 cases of mediastinitis occurred. Of the 8 children with cultures positive for MRSA, 6 developed mediastinitis. The median age for all children with RPI was 32.5 months (n = 108). The median age for children with MRSA and mediastinitis was 6.5 months (n = 8) and 5.5 months (n = 8), respectively. CONCLUSIONS: An alarming increase in the number of RPI cases occurred over the final 4 years. Methicillin-resistant S aureus is now a significant pathogen in patients with RPI at our institution. Documented local increases in community associated MRSA infections and universal sensitivity to clindamycin suggest that community-associated MRSA is responsible for the change in bacteriology. A high correlation exists between MRSA infection and mediastinitis. Patients with MRSA infections are younger and may be vulnerable to developing mediastinitis because of immature immune systems. A higher index of suspicion is needed for MRSA, especially in patients younger than 1 year. PMID- 18427008 TI - The role of airway fluoroscopy in the evaluation of children with stridor. AB - OBJECTIVE: To determine the sensitivity and specificity of airway fluoroscopy in the diagnosis of pediatric laryngotracheal abnormalities. DESIGN: Retrospective chart review. SETTING: Tertiary care children's hospital. PATIENTS: Thirty-nine children, with a mean age of 18 months at the time of evaluation, were evaluated for stridor. MAIN OUTCOME MEASURES: Diagnoses made by airway fluoroscopy and endoscopy. The medical records of patients who had undergone both airway fluoroscopy and airway endoscopy for the evaluation of stridor over a 5-year period were reviewed. The sensitivity, specificity, and positive and negative predictive values of airway fluoroscopy in the diagnosis of laryngomalacia, tracheomalacia, airway stenosis at any level, and airway mass lesions were determined using endoscopic evaluation as the "gold standard." RESULTS: Twenty three of 39 patients (59%) received a different diagnosis by airway endoscopy than by airway fluoroscopy. The sensitivity of airway fluoroscopy in the diagnosis of laryngomalacia, tracheomalacia, airway stenosis, and an airway mass was 27%, 20%, 69%, and 43%, respectively. The specificity for the same diagnoses was 100%, 94%, 100%, and 100%, respectively. CONCLUSIONS: Airway fluoroscopy appears to be reliable in the diagnosis of laryngomalacia, tracheomalacia, airway stenosis, and airway masses because of its high specificity. However, its sensitivity in detecting these common causes of stridor is poor. Negative fluoroscopic study results require further diagnostic evaluation if the clinical indication exists; therefore, the value of fluoroscopy as a screening tool remains uncertain. PMID- 18427009 TI - Effect of septoplasty on inferior turbinate hypertrophy. AB - OBJECTIVE: To measure the effect of septoplasty on the volume of inferior turbinate in patients with a deviated nasal septum. DESIGN: In this retrospective analysis, patients who underwent septoplasty without turbinate surgery from May 1, 2003, through April 30, 2006, were studied. The thicknesses and cross sectional areas of mucosa and conchal bones were measured with computed tomography before the operations and at least 1 year after the operations. SETTING: University hospital. PATIENTS: A total of 20 patients who presented with a chief concern of nasal obstruction. MAIN OUTCOME MEASURES: The thicknesses of the medial mucosa, bone, and lateral mucosa and the cross-sectional area of turbinate before and after septoplasty were compared using the Wilcoxon signed rank test. P < .05 was considered statistically significant. RESULTS: The medial mucosa and cross-sectional area of the inferior turbinate on the concave side of the septum were significantly decreased by septoplasty (both, P = .01), and the medial mucosa and cross-sectional area of the inferior turbinate on the convex side of the septum were significantly increased by septoplasty (P = .01). The thicknesses and cross-sectional areas of the conchal bone on the concave and convex sides of the septum were not affected by septoplasty. CONCLUSION: After septoplasty, inferior turbinate hypertrophy, especially in the medial mucosa, may reverse. PMID- 18427010 TI - Infection rate and virus-induced cytokine secretion in experimental rhinovirus infection in mucosal organ culture: comparison between specimens from patients with chronic rhinosinusitis with nasal polyps and those from normal subjects. AB - OBJECTIVE: To investigate the difference in susceptibility to rhinovirus (RV) infection and RV-induced inflammatory response between the nasal mucosae from patients with chronic rhinosinusitis with nasal polyps (CRS/NP) and subjects without CRS/NP (hereinafter, normal subjects). DESIGN: In vitro study. SETTING: Tertiary care rhinology clinic. PATIENTS: We conducted RV infection experiments on the organ cultures of NPs and inferior turbinate mucosae from 16 patients with CRS/NP and sphenoid sinus and inferior turbinate mucosae from 19 patients who underwent transsphenoidal pituitary surgery. MAIN OUTCOME MEASURES: Successful RV 16 infection was determined by positive identification of RV on the surface fluid of organ culture using seminested reverse transcriptase-polymerase chain reaction. Effects of RV on interleukin 6 (IL-6) and IL-8 secretion were measured by enzyme-linked immunosorbent assay. RESULTS: The successful RV infection was achievable in 9 of 16 NP samples (56.3%) and 9 of 16 turbinate samples (56.3%) from patients with CRS/NP compared with 11 of 19 sphenoid sinus samples (57.9%) and 15 of 19 turbinate samples (78.9%) from normal subjects. The RV infection increased IL-6 and IL-8 secretion 236% and 173%, respectively, in NP samples, and 218% and 178%, respectively, in turbinate samples from patients with CRS/NP; compared with 231% and 145%, respectively, in sphenoid mucosa samples, and 181% and 148%, respectively, in turbinate samples from normal subjects. However, there were no statistical differences among the 4 groups. CONCLUSION: These in vitro findings suggest that subjects with CRS/NP mucosa might not be more susceptible to RV infection, and did not secrete more cytokines in response to rhinovirus infection, than those with normal mucosa. PMID- 18427011 TI - Improvement in quality of life after nasal surgery alone for patients with obstructive sleep apnea and nasal obstruction. AB - OBJECTIVE: To evaluate the impact of nasal surgery alone on quality of life (QOL) in patients with obstructive sleep apnea and nasal obstruction using generic and disease-specific QOL questionnaires. DESIGN: Prospective, longitudinal cohort study. PATIENTS: Fifty-one consecutive patients with obstructive sleep apnea (50 men and 1 woman; mean age, 39 years; mean [SD] apnea-hypopnea index, 37.4 [28.9] events/h; and mean +/- SD body mass index [calculated as weight in kilograms divided by height in meters squared], 26.0 [3.5]) with symptoms of nasal obstruction due to a deviated nasal septum. INTERVENTION: Septomeatoplasty. OUTCOME MEASURES: Surgical outcomes were measured using the Snore Outcomes Survey, the Epworth Sleepiness Scale, and the Medical Outcomes Study 36-Item Short-Form Health Survey (SF-36) 3 months after surgery. We compared baseline and postoperative scores. Normative SF-36 data obtained from 4591 age- and sex matched adults were used as references. RESULTS: Nasal obstruction symptoms significantly improved (mean [SD] visual analog scale score, -5.2 [1.4]; P < .001). Assessments also showed significant improvement in the Snore Outcomes Survey (P < .001) and Epworth Sleepiness Scale (P < .001) scores and 6 of the 8 SF-36 subscale scores (P < .05). Remarkable improvements were observed in disease specific Snore Outcomes Survey (by 43.1%), Epworth Sleepiness Scale (by 27.3%), and generic SF-36 role-emotional (by 30.4%) and role-physical (by 20.7%) QOL subscales. The postoperative role-emotional, bodily pain, and social function dimensions of health were indistinguishable from referential population data (P > .05). CONCLUSIONS: Correction of an obstructed nasal airway significantly improves disease-specific and generic QOL in adult patients with obstructive sleep apnea who also have nasal obstruction symptoms. After nasal surgery, patients may experience greater improvement in snoring and daytime sleepiness than in other generic health status. Our findings substantiate the role of nasal surgery in treating patients with obstructive sleep apnea and nasal obstruction. PMID- 18427012 TI - Adolescent tracheobronchomalacia: double aortic arches revisited. PMID- 18427013 TI - Proliferative myositis: a rare pseudomalignant tumor of the head and neck. PMID- 18427014 TI - Radiology quiz case 1. Anaplastic thyroid carcinoma. PMID- 18427015 TI - Radiology quiz case 2. Pseudoaneurysm of the facial artery. PMID- 18427016 TI - Pathology quiz case 1. Oncocytic lipoadenoma of the parotid gland. PMID- 18427017 TI - Pathology quiz case 2. Atypical meningioma with clear cell features (World Health Organization [WHO] grade II). PMID- 18427018 TI - Natural orifice transluminal endoscopic surgery: lessons learned from the laparoscopic revolution. PMID- 18427019 TI - Surgery for hyperparathyroidism in image-negative patients. AB - HYPOTHESIS: Patients with primary hyperparathyroidism and negative preoperative localization imaging have a different outcome than patients with positive imaging. DESIGN: Prospective single-surgeon case series. SETTING: Referral center. PATIENTS: Forty-two patients with primary hyperparathyroidism, indications for surgery, and both cervical ultrasonographic results and technetium Tc 99m sestamibi nuclear images that were nonlocalizing over a 5- year span. MAIN OUTCOME MEASURES: Extent of surgery required to produce cure; operative findings. RESULTS: Of 430 patients undergoing surgery for primary hyperparathyroidism, 351 underwent both ultrasonographic and sestamibi imaging. Among 351 patients, the imaging results of 42 patients did not show an adenoma, and these patients underwent cervical exploratory surgery. Of 42 patients, 41 were cured at a mean follow-up of 90 days; 1 patient underwent surgical reexploration and was cured by removal of a mediastinal adenoma. To achieve initial cure, 12 of 42 patients (28.6%) required partial thyroidectomy, 9 (21.4%) required partial thymectomy, 17 (40.5%) required paratracheal dissection to access or devascularize an obscure adenoma. Pathologic examination disclosed single adenoma in 26 of 42 patients (61.9%), parathyroid hyperplasia in 14 (33.3%), and double adenoma in 2 (4.8%). CONCLUSIONS: Patients whose preoperative localization studies fail to localize solitary adenoma commonly require extensive surgery to cure hyperparathyroidism. Lack of localization may be a reasonable criterion on which to base referral of the patient to a high-volume medical center. PMID- 18427020 TI - Volume-outcome relationship for coronary artery bypass grafting in an era of decreasing volume. AB - HYPOTHESIS: We hypothesized that the recent reduction in procedure volume for coronary artery bypass grafting (CABG) has led to an increase in the in-hospital mortality rate. DESIGN: Hospital discharge data from the Nationwide Inpatient Sample from January 1, 1988, through December 31, 2003. SETTING: A 20% random sample of patients admitted to US hospitals. PATIENTS: All patients who underwent CABG or percutaneous transluminal coronary interventions. Facilities performing CABG were assigned to standard volume cutoffs. MAIN OUTCOME MEASURES: Rates of cardiac procedures and the proportion of hospitals meeting standard volume cutoffs, as well as the CABG mortality rate. RESULTS: During our 16-year study period, the rate of CABG increased from 7.2 cases per 1000 discharges in 1988 to 12.2 cases in 1997 but then decreased to 9.1 cases in 2003, while the rate of percutaneous interventions tripled. For CABG, the proportion of high-volume hospitals declined from 32.5% in 1997 to 15.5% in 2003. Despite shifts between high- and low-volume hospitals, the CABG mortality rate steadily declined from 5.4% in 1988 to 3.3% in 2003. Hospitals performing the lowest volume of CABG experienced the largest decrease in the in-hospital mortality rate. CONCLUSIONS: Since 1997, CABG volume has declined in the setting of a decrease in in-hospital mortality. A lower mortality rate in the setting of reduced CABG volume is a counterintuitive finding, suggesting that procedure volume is an insufficient predictor of outcome on which to base regionalization strategies. PMID- 18427021 TI - A longitudinal analysis of the general surgery workforce in the United States, 1981-2005. AB - HYPOTHESIS: The overall supply of general surgeons per 100 000 population has declined in the past 2 decades, and small and isolated rural areas of the United States continue to have relatively fewer general surgeons per 100 000 population than urban areas. DESIGN: Retrospective longitudinal analysis. SETTING: Clinically active general surgeons in the United States. PARTICIPANTS: The American Medical Association's Physician Masterfiles from 1981, 1991, 2001, and 2005 were used to identify all clinically active general surgeons in the United States. MAIN OUTCOME MEASURES: Number of general surgeons per 100 000 population and the age, sex, and locale of these surgeons. RESULTS: General surgeon to population ratios declined steadily across the study period, from 7.68 per 100 000 in 1981 to 5.69 per 100 000 in 2005. The overall urban ratio dropped from 8.04 to 5.85 (-27.24%) across the study period, and the overall rural ratio dropped from 6.36 to 5.02 (-21.07%). The average age of rural surgeons increased compared with their urban counterparts, and women were disproportionately concentrated in urban areas. CONCLUSIONS: The overall number of general surgeons per 100 000 population has declined by 25.91% during the past 25 years. The decline has been most marked in urban areas. However, more remote rural areas continue to have significantly fewer general surgeons per 100 000 population. These findings have implications for training, recruiting, and retaining general surgeons. PMID- 18427022 TI - First-line chemotherapy vs bowel tumor resection plus chemotherapy for patients with unresectable synchronous colorectal hepatic metastases. AB - HYPOTHESIS: Bowel resection followed by chemotherapy is a better management strategy than immediate chemotherapy in asymptomatic patients with colorectal cancer and unresectable liver-only metastases at presentation. DESIGN: Retrospective study. SETTING: University hospital. PATIENTS: Sixty-five consecutive symptom-free colorectal cancer patients with unresectable synchronous metastases confined to the liver undergoing bowel tumor resection plus systemic chemotherapy (42 patients [resection group]) or chemotherapy first (23 patients [chemotherapy group]). MAIN OUTCOME MEASURES: Long-term survival and identification of prognostic indicators of outcome. RESULTS: In the resection group, the mean and median overall survival times were shown to be significantly better than those in the chemotherapy group (P = .03). Performance status, basal serum levels of lactic dehydrogenase and alkaline phosphatase, percentage of liver involvement, potentially curative resection of the bowel tumor, and type of treatment (resection vs chemotherapy) were demonstrated to be the only variables significantly correlated with long-term survival. On multivariate analysis, performance status, extent of liver involvement, and type of treatment were shown to be the only covariates independently associated with survival rate. The rate of liver metastasis downstaging with subsequent curative hepatic resection was clearly associated with good performance status, limited liver involvement, and resection of the bowel tumor. CONCLUSIONS: Achieving complete cure in asymptomatic colorectal cancer patients with unresectable synchronous liver-only metastases appears to be mostly the result of shrinkage and resection of hepatic metastases. In patients with good performance status and limited liver involvement, bowel tumor resection appears to be the best treatment option for this purpose. PMID- 18427023 TI - Complications in antireflux surgery: national-based analysis of laparoscopic and open fundoplications. AB - HYPOTHESIS: Longer experience of surgeons has reduced the rate of complications in antireflux surgery. DESIGN: Comparison of the rate of serious complications between open and laparoscopic fundoplication in Finland at the national level. SETTING: University teaching hospital. PATIENTS: From January 1, 1992, to December 31, 2001, 10 846 fundoplications were performed in Finland. Of these, 3987 (37%) were open and 6859 (63%) were laparoscopic. MAIN OUTCOME MEASURES: Administrative databases provided the number of fundoplications, the rate of severe complications, and the mortality. Medical records allowed for evaluation of the nature and cause of severe complications of laparoscopic and open fundoplications. RESULTS: From January 1, 1992, to December 31, 2001, hospital mortality was significantly lower after laparoscopy (P = .01). In comparable groups, surgical mortality or the overall rate of serious complications did not differ. The rate of serious complications decreased after both open surgery (P = .01) and laparoscopic surgery (P = .03). After laparoscopy, patients made claims for injuries more often (P = .003) and had a higher rate of dysphagia (P < .001). In all of the patients with severe dysphagia or fundic perforations after laparoscopy, the short gastric vessels were not divided. Furthermore, 1 open fundoplication and 22 laparoscopic fundoplications had to have reoperations performed owing to dysphagia, mostly involving technical failure. CONCLUSIONS: At the national level, the first 10-year experience of laparoscopic fundoplication reduced the rate of serious complications. The complications largely were technical failures related to the lack of a standardized surgical technique. PMID- 18427024 TI - Porcine and bovine surgical products: Jewish, Muslim, and Hindu perspectives. AB - OBJECTIVE: To determine the acceptability of porcine and bovine surgical implants among persons of Jewish, Muslim, and Hindu faiths whose beliefs prohibit them from consuming porcine and bovine products. DATA SOURCES: An evaluation of current literature concerning religious beliefs among persons of Jewish, Muslim, and Hindu faiths was undertaken to determine if animal-derived surgical implants are permitted for use in these religions. STUDY SELECTION: Because of the limited published literature about this topic, the opinions of religious leaders in Australia were sought. DATA EXTRACTION: Religious and cultural beliefs can conflict with and limit treatment options, especially in surgery. Approximately 81 porcine and bovine surgical implants are regularly used in Australia. DATA SYNTHESIS: It is deemed acceptable for members of the Jewish faith to undergo surgery using porcine products. In dire situations and only after all other options have been exhausted, followers of the Muslim faith are permitted to use porcine surgical products. Hindu religious leaders did not accept the use of bovine surgical implants. CONCLUSIONS: Australia comprises a multicultural society; therefore, it is necessary to consider religious beliefs of all patients. As part of a surgeon's duty of care, the informed consent process should include a discussion about animal-derived surgical implants to avoid religious distress and possible litigation. A greater understanding of religious views would enhance the medical care of persons of Jewish, Muslim, and Hindu faiths. PMID- 18427025 TI - Randomized clinical trial of small-incision and laparoscopic cholecystectomy in patients with symptomatic cholecystolithiasis: primary and clinical outcomes. AB - OBJECTIVE: To evaluate the primary and clinical outcomes in laparoscopic and small-incision cholecystectomy. DESIGN: Blinded randomized single-center trial emphasizing methodologic quality and generalizability. SETTING: General teaching hospital in the Netherlands. PATIENTS: A total of 257 patients undergoing cholecystectomy for symptomatic cholecystolithiasis. INTERVENTIONS: Laparoscopic cholecystectomy and small-incision cholecystectomy, performed mainly by surgical residents. MAIN OUTCOME MEASURES: Complications and symptom relief were primary outcome measures; conversion rate, operative time, and hospital stay were secondary outcome measures. Feasibility of performing both procedures by residents was evaluated as well. RESULTS: In the 257 patients, surgical residents performed 105 laparoscopic and 118 small-incision cholecystectomies. There were no significant differences in complications, conversion rates, and hospital stay. Operative time was significantly shorter with the small-incision technique. CONCLUSIONS: No differences in primary clinical outcome measures were found between laparoscopic and small-incision cholecystectomy in this randomized trial with emphasis on methodologic quality and generalizability. The gold standard status of laparoscopic cholecystectomy is questionable. Trial Registration isrctn.org Identifier: ISRCTN67485658. PMID- 18427026 TI - Use of vascular clamping in hepatic surgery: lessons learned from 1260 liver resections. AB - HYPOTHESIS: Several techniques have been introduced to minimize intraoperative bleeding in hepatic surgery. Ischemia-reperfusion injuries and intestinal congestion are the main drawbacks of vascular clamping. We hypothesized possible negative effects on early postoperative outcomes associated with different types of vascular clamping during liver resections and evaluated how attitudes have changed in the past 20 years. DESIGN: Retrospective review. SETTING: Academic research institute. PATIENTS: Patients who underwent 1260 consecutive liver resections, 338 of them (26.8%) in patients with cirrhosis. MAIN OUTCOME MEASURES: Postoperative complications and mortality were analyzed relative to liver disease, blood transfusion, vascular clamping, and type of liver resection. RESULTS: Vascular clamping was applied in 594 patients (47.1%). Operative mortality was 4.4% in the vascular clamping group and 2.9% in the nonclamped group, a statistically nonsignificant difference. On multivariate analysis, blood transfusion, major hepatectomies, and the presence of cirrhosis were statistically significantly associated with postoperative complications. Among the overall cohort and among patients with cirrhosis, there was statistically significantly reduced use of vascular clamping and of blood transfusion during the past 20 years. The lowest incidences of severe complications occurred among cases of continuous or hemihepatic clamping. Among 338 patients with cirrhosis, 155 (45.9%) received some type of vascular control; morbidity and mortality rates were similar in the groups with vs those without vascular control. On multivariate analysis, only blood transfusion was statistically significantly associated with postoperative morbidity. Postoperative complications were statistically significantly reduced among patients receiving intermittent compared with continuous clamping. CONCLUSIONS: Vascular clamping can be applied without additional risk during partial hepatectomy. Intermittent or hemihepatic clamping is preferable in patients with cirrhosis. PMID- 18427027 TI - Patient and physician preferences for surgical and adjuvant treatment options for rectal cancer. AB - HYPOTHESIS: Patients and their clinicians hold varying preferences for surgical and adjuvant treatment therapies for rectal cancer. DESIGN: Preferences were determined using the Prospective Measure of Preference. SETTING: Royal Prince Alfred and St Vincent's hospitals in Sydney, Australia. PARTICIPANTS: Patients with colorectal cancer were interviewed during their postoperative hospital stay, and physicians were asked to complete a mailed survey. MAIN OUTCOME MEASURES: The Prospective Measure of Preference method produces 2 outcome measures of preference: willingness to trade and prospective measure of preference time trade off. RESULTS: Patients' strongest preference was to avoid a stoma: more than 60% would give up a mean of 34% of their life expectancy to avoid this surgical option. This was followed by treatment options involving chemoradiotherapy, where more than 50% would give up a mean of almost 25% of their life to avoid treatment. Surgeons held stronger preferences against all adjuvant options compared with oncologists (P or=3) varied with headache type. Prevalence was significantly higher in transformed migraine (TM, 68.3%) than in episodic migraine (63.2%, p < 0.01) and significantly elevated in both of these groups compared with probable migraine (42.6%), other chronic daily headaches (36.8%), and severe episodic tension-type headache (36.7%). The prevalence of severe CA (ASC score >or=9) was also highest in TM (28.5%) followed by migraine (20.4%), probable migraine (12.3%), other chronic daily headaches (6.2%), and severe episodic tension-type headache (5.1%). In the migraine and TM groups, prevalence of CA was higher in women and increased with disability score. Among migraineurs, CA increased with headache frequency and body mass index. In all groups, ASC scores were higher in individuals with major depression. CONCLUSIONS: Cutaneous allodynia (CA) is more common and more severe in transformed migraine and migraine than in other primary headaches. Among migraineurs, CA is associated with female sex, headache frequency, increased body mass index, disability, and depression. PMID- 18427072 TI - Acute treatment and prevention of menstrually related migraine headache: evidence based review. AB - Menstrually related migraine (MRM) headache is common in women and associated with substantial disability. Compared to nonmenstrual migraine, MRM attacks are more severe, longer in duration, and have a poorer response to analgesics. The purpose of this guideline is to provide a systematic review and meta-analysis of the existing therapy trials for MRM and evidence-based recommendations for acute and short-term preventive treatment of MRM headache. Prospective, double-blind, randomized controlled trials of any pharmacologic agent for the symptomatic relief or prevention of MRM headache were included in the guideline. The main outcomes considered were the pain response and pain-free response at 2 hours for acute treatment trials, and the incidence of MRM or the number of days on which MRM attacks occurred for short-term prevention trials. Nineteen trials were included in the analysis. The US Preventive Services Task Force quality criteria were used to assess trial quality and to grade recommendations. Based on the evidence, grade B recommendations can be made for the use of sumatriptan 50 and 100 mg, mefenamic acid 500 mg, and rizatriptan 10 mg for the acute treatment of MRM. For the preventive treatment of MRM, there are grade B recommendations for the perimenstrual use of transcutaneous estrogen 1.5 mg, frovatriptan 2.5 mg twice daily, and naratriptan 1 mg twice daily. Choosing among treatment strategies must be based on clinical considerations. PMID- 18427073 TI - Invited article: Managing disruptive physician behavior: impact on staff relationships and patient care. AB - Disruptive behavior can have a significant impact on care delivery, which can adversely affect patient safety and quality outcomes of care. Disruptive behavior occurs across all disciplines but is of particular concern when it involves physicians and nurses who have primary responsibility for patient care. There is a higher frequency of disruptive behavior in neurologists compared to most other nonsurgical specialties. Disruptive behavior causes stress, anxiety, frustration, and anger, which can impede communication and collaboration, which can result in avoidable medical errors, adverse events, and other compromises in quality care. Health care organizations need to be aware of the significance of disruptive behaviors and develop appropriate policies, standards, and procedures to effectively deal with this serious issue and reinforce appropriate standards of behavior. Having a better understanding of what contributes to, incites, or provokes disruptive behaviors will help organizations provide appropriate educational and training programs that can lessen the likelihood of occurrence and improve the overall effectiveness of communication among the health care team. PMID- 18427074 TI - Reduced RAGE mRNA in mononuclear blood cells of patients with probable Alzheimer disease. PMID- 18427075 TI - Schwannomatosis. PMID- 18427076 TI - Re: Brain SPECT perfusion of frontotemporal dementia associated with motor neuron disease. PMID- 18427077 TI - Diagnostic pitfalls in sporadic transthyretin familial amyloid polyneuropathy (TTR-FAP). PMID- 18427079 TI - Diagnosing the aging physician. PMID- 18427078 TI - Ending life with grace and agreement. PMID- 18427080 TI - Global Health forum urges training programs. PMID- 18427081 TI - Abramson: direct-to-consumer advertising will erode health care. PMID- 18427082 TI - Ditch that white coat. PMID- 18427084 TI - Proposed cuts to sport and recreation could hinder health of northern communities. PMID- 18427086 TI - Emergency battlefield cricothyrotomy. PMID- 18427087 TI - Angioedema of the tongue. PMID- 18427088 TI - Black hairy tongue: what is your call? PMID- 18427089 TI - The OPALS Major Trauma Study: impact of advanced life-support on survival and morbidity. AB - BACKGROUND: To date, the benefit of prehospital advanced life-support programs on trauma-related mortality and morbidity has not been established METHODS: The Ontario Prehospital Advanced Life Support (OPALS) Major Trauma Study was a before after systemwide controlled clinical trial conducted in 17 cities. We enrolled adult patients who had experienced major trauma in a basic life-support phase and a subsequent advanced life-support phase (during which paramedics were able to perform endotracheal intubation and administer fluids and drugs intravenously). The primary outcome was survival to hospital discharge. RESULTS: Among the 2867 patients enrolled in the basic life-support (n = 1373) and advanced life-support (n = 1494) phases, characteristics were similar, including mean age (44.8 v. 47.5 years), frequency of blunt injury (92.0% v. 91.4%), median injury severity score (24 v. 22) and percentage of patients with Glasgow Coma Scale score less than 9 (27.2% v. 22.1%). Survival did not differ overall (81.1% among patients in the advanced life-support phase v. 81.8% among those in the basic life-support phase; p = 0.65). Among patients with Glasgow Coma Scale score less than 9, survival was lower among those in the advanced life-support phase (50.9% v. 60.0%; p = 0.02). The adjusted odds of death for the advanced life-support v. basic life-support phases were nonsignificant (1.2, 95% confidence interval 0.9-1.7; p = 0.16). INTERPRETATION: The OPALS Major Trauma Study showed that systemwide implementation of full advanced life-support programs did not decrease mortality or morbidity for major trauma patients. We also found that during the advanced life-support phase, mortality was greater among patients with Glasgow Coma Scale scores less than 9. We believe that emergency medical services should carefully re-evaluate the indications for and application of prehospital advanced life support measures for patients who have experienced major trauma. PMID- 18427090 TI - Effect of mechanical ventilation in the prone position on clinical outcomes in patients with acute hypoxemic respiratory failure: a systematic review and meta analysis. AB - BACKGROUND: Mechanical ventilation in the prone position is used to improve oxygenation in patients with acute hypoxemic respiratory failure. We sought to determine the effect of mechanical ventilation in the prone position on mortality, oxygenation, duration of ventilation and adverse events in patients with acute hypoxemic respiratory failure. METHODS: In this systematic review we searched MEDLINE, EMBASE, the Cochrane Central Register of Controlled Trials and Science Citation Index Expanded for articles published from database inception to February 2008. We also conducted extensive manual searches and contacted experts. We extracted physiologic data and clinically relevant outcomes. RESULTS: Thirteen trials that enrolled a total of 1559 patients met our inclusion criteria. Overall methodologic quality was good. In 10 of the trials (n = 1486) reporting this outcome, we found that prone positioning did not reduce mortality among hypoxemic patients (risk ratio [RR] 0.96, 95% confidence interval [CI] 0.84-1.09; p = 0.52). The lack of effect of ventilation in the prone position on mortality was similar in trials of prolonged prone positioning and in patients with acute lung injury. In 8 of the trials (n = 633), the ratio of partial pressure of oxygen to inspired fraction of oxygen on day 1 was 34% higher among patients in the prone position than among those who remained supine (p < 0.001); these results were similar in 4 trials on day 2 and in 5 trials on day 3. In 9 trials (n = 1206), the ratio in patients assigned to the prone group remained 6% higher the morning after they returned to the supine position compared with patients assigned to the supine group (p = 0.07). Results were quantitatively similar but statistically significant in 7 trials on day 2 and in 6 trials on day 3 (p = 0.001). In 5 trials (n = 1004), prone positioning was associated with a reduced risk of ventilator-associated pneumonia (RR 0.81, 95% CI 0.66-0.99; p = 0.04) but not with a reduced duration of ventilation. In 6 trials (n = 504), prone positioning was associated with an increased risk of pressure ulcers (RR 1.36, 95% CI 1.07 1.71; p = 0.01). Most analyses found no to moderate between-trial heterogeneity. INTERPRETATION: Mechanical ventilation in the prone position does not reduce mortality or duration of ventilation despite improved oxygenation and a decreased risk of pneumonia. Therefore, it should not be used routinely for acute hypoxemic respiratory failure. However, a sustained improvement in oxygenation may support the use of prone positioning in patients with very severe hypoxemia, who have not been well-studied to date. PMID- 18427092 TI - Should invasive airway management be done in the field? PMID- 18427091 TI - Traumatic brain injury: can the consequences be stopped? AB - Traumatic brain injury is a leading cause of morbidity and death in both industrialized and developing countries. To date, there is no targeted pharmacological treatment that effectively limits the progression of secondary injury. The delayed progression of deterioration of grey and white matter gives hope that a meaningful intervention can be applied in a realistic timeframe following initial trauma. In this review we discuss new insights into the subcellular mechanisms of secondary injury that have highlighted numerous potential targets for intervention. PMID- 18427093 TI - Ventilation in the prone position: for some but not for all? PMID- 18427094 TI - Health geography: supporting public health policy and planning. PMID- 18427095 TI - Serious adverse events in academic critical care research. PMID- 18427097 TI - Effectiveness of selective serotonin reuptake inhibitors. PMID- 18427098 TI - Health Check program. PMID- 18427100 TI - A "take with a grain of salt" label for Holiday Review articles. PMID- 18427101 TI - Health Check program. PMID- 18427102 TI - Health Check program. PMID- 18427103 TI - Health Check program. PMID- 18427104 TI - Health Check program. PMID- 18427105 TI - Were polio to return, today... PMID- 18427106 TI - Notch signaling mediates hypoxia-induced tumor cell migration and invasion. AB - Tumor hypoxia is linked to increased metastatic potential, but the molecular mechanisms coupling hypoxia to metastasis are poorly understood. Here, we show that Notch signaling is required to convert the hypoxic stimulus into epithelial mesenchymal transition (EMT), increased motility, and invasiveness. Inhibition of Notch signaling abrogated hypoxia-induced EMT and invasion, and, conversely, an activated form of Notch could substitute for hypoxia to induce these processes. Notch signaling deploys two distinct mechanisms that act in synergy to control the expression of Snail-1, a critical regulator of EMT. First, Notch directly up regulated Snail-1 expression by recruitment of the Notch intracellular domain to the Snail-1 promoter, and second, Notch potentiated hypoxia-inducible factor 1alpha (HIF-1alpha) recruitment to the lysyl oxidase (LOX) promoter and elevated the hypoxia-induced up-regulation of LOX, which stabilizes the Snail-1 protein. In sum, these data demonstrate a complex integration of the hypoxia and Notch signaling pathways in regulation of EMT and open up perspectives for pharmacological intervention with hypoxiainduced EMT and cell invasiveness in tumors. PMID- 18427107 TI - Dendritic cells in islets of Langerhans constitutively present beta cell-derived peptides bound to their class II MHC molecules. AB - Islets of Langerhans from normal mice contained dendritic cells (DCs) in the range of 8-10 per islet. DCs were found in several mouse strains, including those from lymphocyte-deficient mice. DCs were absent in islets from colony stimulating factor-1 deficient mice and this absence correlated with small size islets. Most DCs were found next to blood vessels and resided in islets for several days. Some DCs contained insulin-like granules, and most expressed peptide-MHC complexes derived from beta cell proteins. Islet DCs were highly effective in presenting beta cell antigens to CD4 T cells ex vivo. Presentation of beta cell-derived peptide-MHC complexes by DCs neither depended on islet inflammation nor correlated with the extent of spontaneous beta cell death. Periislet stroma DCs did not contain beta cell peptide-MHC complexes; however, 50% of DCs in pancreatic node were positive. Hence, presentation of high levels of beta cell antigens normally takes place by islet DCs, a finding that has to be placed in the perspective of autoimmune diabetes. PMID- 18427108 TI - The molecular and cellular basis of olfactory-driven behavior in Anopheles gambiae larvae. AB - The mosquito Anopheles gambiae is the principal Afrotropical vector for human malaria. A central component of its vectorial capacity is the ability to maintain sufficient populations of adults. During both adult and preadult (larval) stages, the mosquitoes depend on the ability to recognize and respond to chemical cues that mediate feeding and survival. In this study, we used a behavioral assay to identify a range of odorant-specific responses of An. gambiae larvae that are dependent on the integrity of the larval antennae. Parallel molecular analyses have identified a subset of the An. gambiae odorant receptors (AgOrs) that are localized to discrete neurons within the larval antennae and facilitate odor evoked responses in Xenopus oocytes that are consistent with the larval behavioral spectrum. These studies shed light on chemosensory-driven behaviors and represent molecular and cellular characterization of olfactory processes in mosquito larvae. These advances may ultimately enhance the development of vector control strategies, targeting olfactory pathways in larval-stage mosquitoes to reduce the catastrophic effects of malaria and other diseases. PMID- 18427109 TI - A polyketide macrolactone synthase from the filamentous fungus Gibberella zeae. AB - Resorcylic acid lactones represent a unique class of fungal polyketides and display a wide range of biological activities, such as nanomolar inhibitors of Hsp90 and MAP kinase. The biosynthesis of these compounds is proposed to involve two fungal polyketide synthases (PKS) that function collaboratively to yield a 14 membered macrolactone with a resorcylate core. We report here the reconstitution of Gibberella zeae PKS13, which is the nonreducing PKS associated with zearalenone biosynthesis. Using a small molecule mimic of the natural hexaketide starter unit, we reconstituted the entire repertoire of PKS13 activities in vitro, including starter-unit selection, iterative condensation, regioselective C2-C7 cyclization, and macrolactone formation. PKS13 synthesized both natural 14 membered and previously uncharacterized 16-membered resorcylic acid lactones, indicating relaxed control in both iterative elongation and macrocyclization. PKS13 exhibited broad starter-unit specificities toward fatty acyl-CoAs ranging in sizes between C6 and C16 and displayed the highest activity toward decanoyl CoA. PKS13 was shown to be active in Escherichia coli and synthesized numerous alkyl pyrones and alkyl resorcylic esters without exogenously supplied precursors. We demonstrated that PKS13 can interact with E. coli fatty acid biosynthetic machinery and can be primed with fatty-acyl ACPp at low-micromolar concentrations. PKS13 synthesized new polyketides in E. coli when the culture was supplemented with synthetic precursors, showcasing its utility in precursor directed biosynthesis. PKS13 is therefore a highly versatile polyketide macrolactone synthase that is useful in the engineered biosynthesis of polyketides, including resorcylic acid lactones that are not found in nature. PMID- 18427111 TI - Strong human association with plant invasion success for Trifolium introductions to New Zealand. AB - It has proven difficult, when focused only on biological determinants, to explain why some plant species become naturalized in or invade new locations, whereas others fail. We analyzed the invasion of Trifolium (true clover) species into New Zealand, assessing a range of human, biogeographic, and biological influences at three key invasion stages: introduction, naturalization, and spread. We used sparse principal component analysis (SPCA) to define suites of related attributes and aggregated boosted trees to model relationships with invasion outcomes. Human and biogeographic attributes were strongly associated with success at all stages. Whereas biogeographic attributes, notably large native range, were consistently associated with success, different human factors appeared to favor success at different stages, such as presence in early trade/immigration hotspots (introduction), intentional large-scale planting (naturalization), and frequent presence as a seed contaminant (relative spread rate). Biological traits were less strongly associated with success for introduction and spread and little if at all for naturalization; we found that tall perennials with long flowering periods were more frequently selected for introduction, whereas species with extended flowering in New Zealand spread more rapidly. In addition to causal relationships, the importance of human factors may reflect indirect associations, including ecological traits associated with both human use and invasion. Nevertheless, our results highlight key roles that humans can play in facilitating plant invasion via two pathways: (i) commercial introduction leading to widespread planting and concomitant naturalization and spread and (ii) unintentional introduction and spread of species associated with human activities, such as seed contaminants. PMID- 18427110 TI - Antiinflammatory cAMP signaling and cell migration genes co-opted by the anthrax bacillus. AB - Bacillus anthracis, the etiologic agent of anthrax, avoids immune surveillance and commandeers host macrophages as a vehicle for lymphatic spreading. Here, we show that B. anthracis edema toxin (ET), via its adenylyl cyclase activity, dramatically increases the motility of infected macrophages and the expression of vascular endothelial growth factor. The transcription factor CREB and the syndecan-1 gene, a CREB target, play crucial roles in ET-induced macrophage migration. These molecular and cellular responses occur in macrophages engaged in antiinflammatory G protein-coupled receptor activation, thus illustrating a common signaling circuitry controlling resolution of inflammation and host cell hijacking by B. anthracis. PMID- 18427112 TI - Single-cell protein induction dynamics reveals a period of vulnerability to antibiotics in persister bacteria. AB - Phenotypic variability in populations of cells has been linked to evolutionary robustness to stressful conditions. A remarkable example of the importance of cell-to-cell variability is found in bacterial persistence, where subpopulations of dormant bacteria, termed persisters, were shown to be responsible for the persistence of the population to antibiotic treatments. Here, we use microfluidic devices to monitor the induction of fluorescent proteins under synthetic promoters and characterize the dormant state of single persister bacteria. Surprisingly, we observe that protein production does take place in supposedly dormant bacteria, over a narrow time window after the exit from stationary phase. Only thereafter does protein production stop, suggesting that differentiation into persisters fully develops over this time window and not during starvation, as previously believed. In effect, we observe that exposure of bacteria to antibiotics during this time window significantly reduces persistence. Our results point to new strategies to fight persistent bacterial infections. The quantitative measurement of single-cell induction presented in this study should shed light on the processes leading to the dormancy of subpopulations in different systems, such as in subpopulations of viable but nonculturable bacteria, or those of quiescent cancer cells. PMID- 18427113 TI - Calculation of protein-ligand binding free energy by using a polarizable potential. AB - The binding of charged ligands benzamidine and diazamidine to trypsin was investigated by using a polarizable potential energy function and explicit-water molecular dynamics simulations. The binding free energies were computed from the difference between the free energies of decoupling the ligand from water and protein environments. Both the absolute and the relative free energies from the perturbation simulations agree with experimental measurements to within 0.5 kcal.mol(-1). Comparison of free-energy components sampled from different thermodynamic paths indicates that electrostatics is the main driving force behind benzamidine recognition of trypsin. The contribution of electronic polarization to binding appears to be crucial. By computing the free-energy contribution caused by the polarization between the ligand and its surroundings, we found that polarization has the opposite effect in dissimilar environments. Although polarization favors ligand solvation in water, it weakens the protein ligand attraction by screening the electrostatic interaction between trypsin and benzamidine. We also examined the relative binding free energies of a benzamidine analog diazamidine to trypsin. The changes in free energy on benzamidine diazamidine substitution were tens of kilocalories in both water and trypsin environments; however, the change in the total binding free energy is <2 kcal.mol(-1) because of cancellation, consistent with the experimental results. Overall, our results suggest that the use of a polarizable force field, given adequate sampling, is capable of achieving chemical accuracy in molecular simulations of protein-ligand recognition. PMID- 18427114 TI - The mitotic kinesin CENP-E is a processive transport motor. AB - In vivo studies suggest that centromeric protein E (CENP-E), a kinesin-7 family member, plays a key role in the movement of chromosomes toward the metaphase plate during mitosis. How CENP-E accomplishes this crucial task, however, is not clear. Here we present single-molecule measurements of CENP-E that demonstrate that this motor moves processively toward the plus end of microtubules, with an average run length of 2.6 +/- 0.2 mum, in a hand-over-hand fashion, taking 8-nm steps with a stall force of 6 +/- 0.1 pN. The ATP dependence of motor velocity obeys Michaelis-Menten kinetics with K(M,ATP) = 35 +/- 5 muM. All of these features are remarkably similar to those for kinesin-1-a highly processive transport motor. We, therefore, propose that CENP-E transports chromosomes in a manner analogous to how kinesin-1 transports cytoplasmic vesicles. PMID- 18427115 TI - Suppression of interneuron programs and maintenance of selected spinal motor neuron fates by the transcription factor AML1/Runx1. AB - Individual spinal motor neuron identities are specified in large part by the intrinsic repertoire of transcription factors expressed by undifferentiated progenitors and maturing neurons. It is shown here that the transcription factor AML1/Runx1 (Runx1) is expressed in selected spinal motor neuron subtypes after the onset of differentiation and is both necessary and sufficient to suppress interneuron-specific developmental programs and promote maintenance of motor neuron characteristics. These findings show an important role for Runx1 during the consolidation of selected spinal motor neuron identities. Moreover, they suggest a requirement for a persistent suppression of interneuron genes within maturing motor neurons. PMID- 18427116 TI - Neural correlates of mentalizing-related computations during strategic interactions in humans. AB - Competing successfully against an intelligent adversary requires the ability to mentalize an opponent's state of mind to anticipate his/her future behavior. Although much is known about what brain regions are activated during mentalizing, the question of how this function is implemented has received little attention to date. Here we formulated a computational model describing the capacity to mentalize in games. We scanned human subjects with functional MRI while they participated in a simple two-player strategy game and correlated our model against the functional MRI data. Different model components captured activity in distinct parts of the mentalizing network. While medial prefrontal cortex tracked an individual's expectations given the degree of model-predicted influence, posterior superior temporal sulcus was found to correspond to an influence update signal, capturing the difference between expected and actual influence exerted. These results suggest dissociable contributions of different parts of the mentalizing network to the computations underlying higher-order strategizing in humans. PMID- 18427117 TI - Global transcriptomic analysis of Cyanothece 51142 reveals robust diurnal oscillation of central metabolic processes. AB - Cyanobacteria are photosynthetic organisms and are the only prokaryotes known to have a circadian lifestyle. Unicellular diazotrophic cyanobacteria such as Cyanothece sp. ATCC 51142 produce oxygen and can also fix atmospheric nitrogen, a process exquisitely sensitive to oxygen. To accommodate such antagonistic processes, the intracellular environment of Cyanothece oscillates between aerobic and anaerobic conditions during a day-night cycle. This is accomplished by temporal separation of the two processes: photosynthesis during the day and nitrogen fixation at night. Although previous studies have examined periodic changes in transcript levels for a limited number of genes in Cyanothece and other unicellular diazotrophic cyanobacteria, a comprehensive study of transcriptional activity in a nitrogen-fixing cyanobacterium is necessary to understand the impact of the temporal separation of photosynthesis and nitrogen fixation on global gene regulation and cellular metabolism. We have examined the expression patterns of nearly 5,000 genes in Cyanothece 51142 during two consecutive diurnal periods. Our analysis showed that approximately 30% of these genes exhibited robust oscillating expression profiles. Interestingly, this set included genes for almost all central metabolic processes in Cyanothece 51142. A transcriptional network of all genes with significantly oscillating transcript levels revealed that the majority of genes encoding enzymes in numerous individual biochemical pathways, such as glycolysis, oxidative pentose phosphate pathway, and glycogen metabolism, were coregulated and maximally expressed at distinct phases during the diurnal cycle. These studies provide a comprehensive picture of how a physiologically relevant diurnal light-dark cycle influences the metabolism in a photosynthetic bacterium. PMID- 18427118 TI - Compression of auditory space during rapid head turns. AB - Studies of spatial perception during visual saccades have demonstrated compressions of visual space around the saccade target. Here we psychophysically investigated perception of auditory space during rapid head turns, focusing on the "perisaccadic" interval. Using separate perceptual and behavioral response measures we show that spatial compression also occurs for rapid head movements, with the auditory spatial representation compressing by up to 50%. Similar to observations in the visual system, this occurred only when spatial locations were measured by using a perceptual response; it was absent for the behavioral measure involving a nose-pointing task. These findings parallel those observed in vision during saccades and suggest that a common neural mechanism may subserve these distortions of space in each modality. PMID- 18427119 TI - Protein exchange dynamics at chemoreceptor clusters in Escherichia coli. AB - Signal processing in bacterial chemotaxis relies on large sensory complexes consisting of thousands of protein molecules. These clusters create a scaffold that increases the efficiency of pathway reactions and amplifies and integrates chemotactic signals. The cluster core in Escherichia coli comprises a ternary complex composed of receptors, kinase CheA, and adaptor protein CheW. All other chemotaxis proteins localize to clusters by binding either directly to receptors or to CheA. Here, we used fluorescence recovery after photobleaching (FRAP) to investigate the turnover of chemotaxis proteins at the cluster and their mobility in the cytoplasm. We found that cluster exchange kinetics were protein-specific and took place on several characteristic time scales that correspond to excitation, adaptation, and cell division, respectively. We further applied analytical and numerical data fitting to analyze intracellular protein diffusion and to estimate the rate constants of cluster equilibration in vivo. Our results indicate that the rates of protein turnover at the cluster have evolved to ensure optimal performance of the chemotaxis pathway. PMID- 18427120 TI - Bromodomain 4 activation predicts breast cancer survival. AB - Previous work identified the Rap1 GTPase-activating protein Sipa1 as a germ-line encoded metastasis modifier. The bromodomain protein Brd4 physically interacts with and modulates the enzymatic activity of Sipa1. In vitro analysis of a highly metastatic mouse mammary tumor cell line ectopically expressing Brd4 demonstrates significant reduction of invasiveness without altering intrinsic growth rate. However, a dramatic reduction of tumor growth and pulmonary metastasis was observed after s.c. implantation into mice, implying that activation of Brd4 may somehow be manipulating response to tumor microenvironment in the in vivo setting. Further in vitro analysis shows that Brd4 modulates extracellular matrix gene expression, a class of genes frequently present in metastasis-predictive gene signatures. Microarray analysis of the mammary tumor cell lines identified a Brd4 activation signature that robustly predicted progression and/or survival in multiple human breast cancer datasets analyzed on different microarray platforms. Intriguingly, the Brd4 signature also almost perfectly matches a molecular classifier of low-grade tumors. Taken together, these data suggest that dysregulation of Brd4-associated pathways may play an important role in breast cancer progression and underlies multiple common prognostic signatures. PMID- 18427121 TI - A dry ligand-binding cavity in a solvated protein. AB - Ligands usually bind to proteins by displacing water from the binding site. The affinity and kinetics of binding therefore depend on the hydration characteristics of the site. Here, we show that the extreme case of a completely dehydrated free binding site is realized for the large nonpolar binding cavity in bovine beta-lactoglobulin. Because spatially delocalized water molecules may escape detection by x-ray diffraction, we use water (17)O and (2)H magnetic relaxation dispersion (MRD), (13)C NMR spectroscopy, molecular dynamics simulations, and free energy calculations to establish the absence of water from the binding cavity. Whereas carbon nanotubes of the same diameter are filled by a hydrogen-bonded water chain, the MRD data show that the binding pore in the apo protein is either empty or contains water molecules with subnanosecond residence times. However, the latter possibility is ruled out by the computed hydration free energies, so we conclude that the 315 A(3) binding pore is completely empty. The apo protein is thus poised for efficient binding of fatty acids and other nonpolar ligands. The qualitatively different hydration of the beta-lactoglobulin pore and carbon nanotubes is caused by subtle differences in water-wall interactions and water entropy. PMID- 18427122 TI - The antibody zalutumumab inhibits epidermal growth factor receptor signaling by limiting intra- and intermolecular flexibility. AB - The epidermal growth factor receptor (EGFR) activates cellular pathways controlling cell proliferation, differentiation, migration, and survival. It thus represents a valid therapeutic target for treating solid cancers. Here, we used an electron microscopy-based technique (Protein Tomography) to study the structural rearrangement accompanying activation and inhibition of native, individual, EGFR molecules. Reconstructed tomograms (3D density maps) showed a level of detail that allowed individual domains to be discerned. Monomeric, resting EGFR ectodomains demonstrated large flexibility, and a number of distinct conformations were observed. In contrast, ligand-activated EGFR complexes were detected only as receptor dimers with ring-like conformations. Zalutumumab, a therapeutic inhibitory EGFR antibody directed against domain III, locked EGFR molecules into a very compact, inactive conformation. Biochemical analyses showed bivalent binding of zalutumumab to provide potent inhibition of EGFR signaling. The structure of EGFR-zalutumumab complexes on the cell surface visualized by Protein Tomography indicates that the cross-linking spatially separates the EGFR molecules' intracellular kinase domains to an extent that appears incompatible with the induction of signaling. These insights into the mechanisms of action of receptor inhibition may also apply to other cell-surface tyrosine kinase receptors of the ErbB family. PMID- 18427123 TI - Prediction of human errors by maladaptive changes in event-related brain networks. AB - Humans engaged in monotonous tasks are susceptible to occasional errors that may lead to serious consequences, but little is known about brain activity patterns preceding errors. Using functional MRI and applying independent component analysis followed by deconvolution of hemodynamic responses, we studied error preceding brain activity on a trial-by-trial basis. We found a set of brain regions in which the temporal evolution of activation predicted performance errors. These maladaptive brain activity changes started to evolve approximately 30 sec before the error. In particular, a coincident decrease of deactivation in default mode regions of the brain, together with a decline of activation in regions associated with maintaining task effort, raised the probability of future errors. Our findings provide insights into the brain network dynamics preceding human performance errors and suggest that monitoring of the identified precursor states may help in avoiding human errors in critical real-world situations. PMID- 18427124 TI - Higher effect of plant species diversity on productivity in natural than artificial ecosystems. AB - Current and expected changes in biodiversity have motivated major experiments, which reported a positive relationship between plant species diversity and primary production. As a first step in addressing this relationship, these manipulative experiments controlled as many potential confounding covariables as possible and assembled artificial ecosystems for the purpose of the experiments. As a new step in this endeavor, we asked how plant species richness relates to productivity in a natural ecosystem. Here, we report on an experiment conducted in a natural ecosystem in the Patagonian steppe, in which we assessed the biodiversity effect on primary production. Using a plant species diversity gradient generated by removing species while maintaining constant biomass, we found that aboveground net primary production increased with the number of plant species. We also found that the biodiversity effect was larger in natural than in artificial ecosystems. This result supports previous findings and also suggests that the effect of biodiversity in natural ecosystems may be much larger than currently thought. PMID- 18427125 TI - Cardiovascular monitoring of children and adolescents with heart disease receiving medications for attention deficit/hyperactivity disorder [corrected]: a scientific statement from the American Heart Association Council on Cardiovascular Disease in the Young Congenital Cardiac Defects Committee and the Council on Cardiovascular Nursing. PMID- 18427127 TI - Impact of time of presentation on the care and outcomes of acute myocardial infarction. AB - BACKGROUND: Prior studies have demonstrated an inconsistent association between patients' arrival time for acute myocardial infarction (AMI) and their subsequent medical care and outcomes. METHODS AND RESULTS: Using a contemporary national clinical registry, we examined differences in medical care and in-hospital mortality among AMI patients admitted during regular hours (weekdays 7 am to 7 pm) versus off-hours (weekends, holidays, and 7 pm to 7 am weeknights). The study cohort included 62,814 AMI patients from the Get With the Guidelines-Coronary Artery Disease database admitted to 379 hospitals throughout the United States from July 2000 through September 2005. Overall, 33 982 (54.1%) patients arrived during off-hours. Compared with those arriving during regular hours, eligible off hour patients were slightly less likely to receive primary percutaneous coronary intervention (adjusted odds ratio [OR], 0.93; 95% confidence interval [CI], 0.89 to 0.98), had longer door-to-balloon times (median, 110 versus 85 minutes; P<0.0001), and were less likely to achieve door-to-balloon < or = 90 minutes (adjusted OR, 0.34; 95% CI, 0.29 to 0.39). Arrival during off-hours was associated with slightly lower overall revascularization rates (adjusted OR, 0.94; 95% CI, 0.90 to 0.97). No measurable differences, however, were found in in hospital mortality between regular hours and off-hours in the overall AMI, ST elevated MI, and non-ST-elevated MI cohorts (adjusted OR, 0.99; 95% CI, 0.93 to 1.06; adjusted OR, 1.05; 95% CI, 0.94 to 1.18; and adjusted OR, 0.97; 95% CI, 0.90 to 1.04, respectively). Similar observations were made across most age and sex subgroups and with an alternative definition for arrival time (weekends/holidays versus weekdays). CONCLUSIONS: Despite slightly fewer primary percutaneous coronary interventions and overall revascularizations and significantly longer door-to-balloon times, patients presenting with AMI during off-hours had in-hospital mortality similar to those presenting during regular hours. PMID- 18427126 TI - Thymosin beta4 is an essential paracrine factor of embryonic endothelial progenitor cell-mediated cardioprotection. AB - BACKGROUND: Prolonged myocardial ischemia results in cardiomyocyte loss despite successful revascularization. We have reported that retrograde application of embryonic endothelial progenitor cells (eEPCs) provides rapid paracrine protection against ischemia-reperfusion injury. Here, we investigated the role of thymosin beta4 (Tbeta4) as a mediator of eEPC-mediated cardioprotection. METHODS AND RESULTS: In vitro, neonatal rat cardiomyocytes were subjected to hypoxia reoxygenation in the absence or presence of eEPCs with or without Tbeta4 short hairpin RNA (shRNA) transfection. In vivo, pigs (n=9 per group) underwent percutaneous left anterior descending artery occlusion for 60 minutes on day 1. After 55 minutes of ischemia, control eEPCs (5x10(6) cells) or cells transfected with Tbeta4 shRNA when indicated or 15 mg Tbeta4 alone were retroinfused into the anterior interventricular vein. Segmental endocardial shortening in the infarct zone at 150-bpm atrial pacing, infarct size (triphenyl tetrazolium chloride viability and methylene blue exclusion), and inflammatory cell influx (myeloperoxidase activity) were determined 24 hours later. Survival of neonatal rat cardiomyocytes increased from 32+/-4% to 90+/-2% after eEPC application, an effect sensitive to shRNA transfection compared with Tbeta4 (45+/-7%). In vivo, infarct size decreased with eEPC application (38+/-4% versus 54+/-4% of area at risk; P<0.01), an effect abolished by Tbeta4 shRNA (62+/-3%). Segmental subendocardial shortening improved after eEPC treatment (22+/-3% versus -3+/-4% of control area) unless Tbeta4 shRNA was transfected (-6+/-4%). Retroinfusion of Tbeta4 mimicked eEPC application (infarct size, 37+/-3%; segmental endocardial shortening, 34+/-7%). Myeloperoxidase activity (3323+/-388 U/mg in controls) was decreased by eEPCs (1996+/-546 U/mg) or Tbeta4 alone (1455+/-197 U/mg) but not Tbeta4 shRNA-treated eEPCs (5449+/-829 U/mg). CONCLUSIONS: Our findings show that short-term cardioprotection derived by regional application of eEPCs can be attributed, at least in part, to Tbeta4. PMID- 18427128 TI - Mineralocorticoid receptor blockade reverses obesity-related changes in expression of adiponectin, peroxisome proliferator-activated receptor-gamma, and proinflammatory adipokines. AB - BACKGROUND: In obesity, decreases in adiponectin and increases in proinflammatory adipokines are associated with heart disease. Because adipocytes express mineralocorticoid receptor (MR) and MR blockade reduces cardiovascular inflammation and injury, we tested the hypothesis that MR blockade reduces inflammation and expression of proinflammatory cytokines in adipose tissue and increases adiponectin expression in adipose tissue and hearts of obese mice. METHODS AND RESULTS: We determined the effect of MR blockade (eplerenone, 100 mg/kg per day for 16 weeks) on gene expression in retroperitoneal adipose and heart tissue from obese, diabetic db/db mice (n=8) compared with untreated obese, diabetic db/db mice (n=10) and lean, nondiabetic db/+ littermates (n=11). Expression of tumor necrosis factor-alpha, monocyte chemoattractant protein-1, plasminogen activator inhibitor type 1, and macrophage protein CD68 increased, and expression of adiponectin and peroxisome proliferator-activated receptor gamma decreased in retroperitoneal adipose tissue from obese versus lean mice. In addition, adiponectin expression in heart was reduced in obese versus lean mice. MR blockade prevented these obesity-related changes in gene expression. Furthermore, treatment of undifferentiated preadipocytes with aldosterone (10(-8) mol/L for 24 hours) increased mRNA levels of tumor necrosis factor-alpha and monocyte chemoattractant protein-1 and reduced mRNA and protein levels of peroxisome proliferator-activated receptor-gamma and adiponectin, supporting a direct aldosterone effect on gene expression. CONCLUSIONS: MR blockade reduced expression of proinflammatory and prothrombotic factors in adipose tissue and increased expression of adiponectin in heart and adipose tissue of obese, diabetic mice. These effects on adiponectin and adipokine gene expression may represent a novel mechanism for the cardioprotective effects of MR blockade. PMID- 18427129 TI - Noncanonical Wnt11 signaling is sufficient to induce cardiomyogenic differentiation in unfractionated bone marrow mononuclear cells. AB - BACKGROUND: Despite the frequent clinical use of adult unfractionated bone marrow mononuclear cells (BMMNCs) for cardiac repair, whether these cells are capable of undergoing cardiomyogenic differentiation in vitro remains uncertain. In addition, the role of Wnt signaling in cardiomyogenic differentiation of adult cells is unclear. METHODS AND RESULTS: Unfractionated BMMNCs were isolated from adult mice via Ficoll-Paque density-gradient centrifugation and cultured in the presence of Wnt3a or Wnt11. In control BMMNCs, Wnt11 was not expressed, whereas the expression of markers of pluripotency (Oct-4 and Nanog), as well as that of Wnt3a and beta-catenin, decreased progressively during culture. Exposure to Wnt3a rescued beta-catenin expression and markedly increased the expression of Oct-4 and Nanog, concomitant with increased cell proliferation and CD45 expression. In contrast, exposure to ectopically expressed noncanonical Wnt11 markedly decreased the expression of Oct-4 and Nanog and induced mRNA expression (quantitative real time reverse-transcription polymerase chain reaction) of cardiac-specific genes (Nkx2.5, GATA-4, atrial natriuretic peptide, alpha- and beta-myosin heavy chain, and cardiac troponin T) by day 3 with subsequent progression to a pattern characteristic of the cardiac fetal gene program. After 21 days, 27.6+/-0.6% and 29.6+/-1.4% of BMMNCs expressed the cardiac-specific antigens cardiac myosin heavy chain and cardiac troponin T, respectively (immunocytochemistry), indicating cardiomyogenic lineage commitment. Wnt11-induced cardiac-specific expression was completely abolished by the protein kinase C inhibitor bisindolylmaleimide I, partially abolished by the c-Jun-N-terminal kinase inhibitor SP600125, and attenuated by the Wnt inhibitor Dickkopf-1. CONCLUSIONS: In adult density-gradient separated BMMNCs, canonical Wnt3a promotes stemness, proliferation, and hematopoietic commitment, whereas noncanonical signaling via Wnt11 induces robust cardiomyogenic differentiation in a protein kinase C- and c Jun-N-terminal kinase-dependent manner. PMID- 18427130 TI - Depressive symptoms and the risk of atherosclerotic progression among patients with coronary artery bypass grafts. AB - BACKGROUND: Depressive symptoms have been associated with increased risk of coronary artery disease and poor prognosis among patients with existing coronary artery disease, but whether depressive symptoms specifically influence atherosclerotic progression among such patients is uncertain. METHODS AND RESULTS: The Post-CABG Trial randomized patients with a history of coronary bypass graft surgery to either an aggressive or a moderate lipid-lowering strategy and to either warfarin or placebo. Coronary angiography was conducted at enrollment and after a median follow-up of 4.2 years. Depressive symptoms were assessed at enrollment with the Centers for Epidemiologic Studies Depression scale (CES-D) in 1319 patients with 2496 grafts. In models that adjusted for age, gender, race, treatment assignment, and years since coronary bypass graft surgery, a CES-D score > or =16 was positively associated with risk of substantial graft disease progression (OR 1.50, 95% CI 1.08 to 2.10, P=0.02) and marginally associated with a 0.11-mm (95% CI -0.22 to 0.01 mm, P=0.07) decrease in minimum lumen diameter, but not with risk of graft occlusion (P=0.30). Additional adjustment for past medical history, blood pressure, and renal function did not materially alter these results. This association was virtually absent among participants randomly assigned to aggressive lipid-lowering therapy. CONCLUSIONS: These findings suggest that depressive symptoms are associated with a higher risk of atherosclerotic progression among patients with saphenous vein grafts and that aggressive lipid lowering can minimize this increased risk. Whether depressive symptoms increase progression in other types of coronary atherosclerosis and whether aggressive lipid lowering attenuates such progression will require additional study. PMID- 18427131 TI - Effect of short call admission on length of stay and quality of care for acute decompensated heart failure. AB - BACKGROUND: In response to residency work hour restrictions, programs restructured call schedules, increasing the use of short call (daytime admitting teams). Few data exist on the effect of short call on quality of patient care. Our objective was to examine the effect of short call admission on length of stay and quality of care for patients with acute decompensated heart failure. METHODS AND RESULTS: We conducted a retrospective cohort study of 218 patients admitted with acute decompensated heart failure to the Nashville VA Medical Center between July 1, 2003, and June 30, 2005. The primary exposure was short call, and the primary outcome was length of stay. The secondary outcomes--diuretic dosing, weight monitoring, and hospital complications--were determined through a combination of administrative data and chart review. Patients admitted to short call had a longer median length of stay than patients admitted to long call (5.2 days [25% to 75%, 3.2 to 8 days] versus 3.9 days [interquartile range, 2.7 to 6.5 days]; P=0.0004). After adjustment for covariates, short call had a 44% increase in length of stay (95% CI, 15 to 80) compared with long call. Short call patients received fewer diuretic doses in the first 24 hours of hospitalization (1.80 versus 2.12; P=0.014) and had a longer median time to the second dose of loop diuretics compared with long call patients (17.9 hours versus 16.2 hours; P=0.044). CONCLUSIONS: Admission to short call is predictive of increased length of stay, a decreased number of diuretic doses, and delays in the timing of diuretics among patients with acute decompensated heart failure. Additional studies are needed to clarify the impact of short call admission on inpatient quality of care. PMID- 18427132 TI - Cardioprotective and vasodilatory actions of glucagon-like peptide 1 receptor are mediated through both glucagon-like peptide 1 receptor-dependent and -independent pathways. AB - BACKGROUND: The glucagon-like peptide 1 receptor (GLP-1R) is believed to mediate glucoregulatory and cardiovascular effects of the incretin hormone GLP-1(7-36) (GLP-1), which is rapidly degraded by dipeptidyl peptidase-4 (DPP-4) to GLP-1(9 36), a truncated metabolite generally thought to be inactive. Novel drugs for the treatment of diabetes include analogues of GLP-1 and inhibitors of DPP-4; however, the cardiovascular effects of distinct GLP-1 peptides have received limited attention. METHODS AND RESULTS: Here, we show that endothelium and cardiac and vascular myocytes express a functional GLP-1R as GLP-1 administration increased glucose uptake, cAMP and cGMP release, left ventricular developed pressure, and coronary flow in isolated mouse hearts. GLP-1 also increased functional recovery and cardiomyocyte viability after ischemia-reperfusion injury of isolated hearts and dilated preconstricted arteries from wild-type mice. Unexpectedly, many of these actions of GLP-1 were preserved in Glp1r(-/-) mice. Furthermore, GLP-1(9-36) administration during reperfusion reduced ischemic damage after ischemia-reperfusion and increased cGMP release, vasodilatation, and coronary flow in wild-type and Glp1r(-/-) mice, with modest effects on glucose uptake. Studies using a DPP-4-resistant GLP-1R agonist and inhibitors of DPP-4 and nitric oxide synthase showed that the effects of GLP-1(7-36) were partly mediated by GLP-1(9-36) through a nitric oxide synthase-requiring mechanism that is independent of the known GLP-1R. CONCLUSIONS: These data describe cardioprotective actions of GLP-1(7-36) mediated through the known GLP-1R and novel cardiac and vascular actions of GLP-1(7-36) and its metabolite GLP-1(9-36) independent of the known GLP-1R. Our data suggest that the extent to which GLP-1 is metabolized to GLP-1(9-36) may have functional implications in the cardiovascular system. PMID- 18427133 TI - Angiotensin-converting enzyme inhibition improves vascular function in rheumatoid arthritis. AB - BACKGROUND: The excess in cardiovascular risk in patients with rheumatoid arthritis provides a strong rationale for early therapeutical interventions. In view of the similarities between atherosclerosis and rheumatoid arthritis and the proven benefit of angiotensin-converting enzyme inhibitors in atherosclerotic vascular disease, it was the aim of the present study to delineate the impact of ramipril on endothelial function as well as on markers of inflammation and oxidative stress in patients with rheumatoid arthritis. METHODS AND RESULTS: Eleven patients with rheumatoid arthritis were included in this randomized, double-blind, crossover study to receive ramipril in an uptitration design (2.5 to 10 mg) for 8 weeks followed by placebo, or vice versa, on top of standard antiinflammatory therapy. Endothelial function assessed by flow-mediated dilation of the brachial artery, markers of inflammation and oxidative stress, and disease activity were investigated at baseline and after each treatment period. Endothelial function assessed by flow-mediated dilation increased from 2.85+/ 1.49% to 4.00+/-1.81% (P=0.017) after 8 weeks of therapy with ramipril but did not change with placebo (from 2.85+/-1.49% to 2.84+/-2.47%; P=0.88). Although systolic blood pressure and heart rate remained unaltered, diastolic blood pressure decreased slightly from 78+/-7 to 74+/-6 mm Hg (P=0.03). Tumor necrosis factor-alpha showed a significant inverse correlation with flow-mediated dilation (r=-0.408, P=0.02), and CD40 significantly decreased after ramipril therapy (P=0.049). CONCLUSIONS: Angiotensin-converting enzyme inhibition with 10 mg/d ramipril for 8 weeks on top of current antiinflammatory treatment markedly improved endothelial function in patients with rheumatoid arthritis. This finding suggests that angiotensin-converting enzyme inhibition may provide a novel strategy to prevent cardiovascular events in these patients. PMID- 18427134 TI - Long-QT syndrome after age 40. AB - BACKGROUND: Previous studies that assessed the risk of life-threatening cardiac events in patients with congenital long-QT syndrome (LQTS) have focused mainly on the first 4 decades of life, whereas the clinical course of this inherited cardiac disorder in the older population has not been studied. METHODS AND RESULTS: The risk of aborted cardiac arrest or death from age 41 though 75 years was assessed in 2759 subjects from the International LQTS Registry, categorized into electrocardiographically affected (corrected QT interval [QTc] > or = 470 ms), borderline (QTc 440 to 469 ms), and unaffected (QTc < 440 ms) subgroups. The affected versus unaffected adjusted hazard ratio for aborted cardiac arrest or death was 2.65 (P<0.001) in the age range of 41 to 60 years and 1.23 (P=0.31) in the age range of 61 to 75 years. The clinical course of study subjects displayed gender differences: Affected LQTS women experienced a significantly higher cumulative event rate (26%) than borderline (16%) and unaffected (12%) women (P=0.001), whereas event rates were similar among the 3 respective subgroups of men (29%, 26%, and 27%; P=0.16). Recent syncope (< 2 years in the past) was the predominant risk factor in affected subjects (hazard ratio 9.92, P<0.001), and the LQT3 genotype was identified as the most powerful predictor of outcome in a subset of 871 study subjects who were genetically tested for a known LQTS mutation (hazard ratio 4.76, P=0.02). CONCLUSIONS: LQTS subjects maintain a high risk for life-threatening cardiac events after age 40 years. The phenotypic expression of affected subjects is influenced by age-specific factors related to gender, clinical history, and the LQTS genotype. PMID- 18427135 TI - Changes in insulin resistance and cardiovascular risk during adolescence: establishment of differential risk in males and females. AB - BACKGROUND: Developmental changes in insulin resistance and cardiovascular risk were studied in youths 11 to 19 years of age. METHODS AND RESULTS: A cohort was randomly selected after blood pressure screening of Minneapolis, Minn, school children. Studies were done 3 times on this cohort and once on their siblings (996 observations on 507 individuals from 363 families). Insulin sensitivity was determined by euglycemic clamp. Body mass index and waist circumference increased similarly in both sexes from ages 11 to 19 years. Body fat decreased in males and increased in females (P<0.001). Lean body mass increased at a steeper rate in males (P<0.0001). Insulin resistance was lower in males at 11 years but increased steadily to 19 years (P=0.003); in contrast, it did not increase in females. Thus, despite being less insulin resistant at 11 years and decreasing in fatness during puberty, males became more insulin resistant than females by 19 years of age. Triglycerides increased in males and high-density lipoprotein cholesterol decreased, whereas the opposite pattern was seen in females, which resulted in higher triglycerides and lower high-density lipoprotein cholesterol in males at 19 years. No gender difference in low-density lipoprotein or total cholesterol was seen. Systolic blood pressure increased in both sexes but at a greater rate in boys (P=0.03). CONCLUSIONS: During the transition from late childhood through adolescence, insulin resistance in males increased in association with increased triglycerides and decreased high-density lipoprotein cholesterol, despite a concurrent reduction in body fatness, whereas the opposite occurred in females. These gender-related developmental changes in insulin resistance, which were independent from changes in fatness, total cholesterol, and low-density lipoprotein cholesterol, are consistent with an early role for insulin resistance in the increased cardiovascular risk found in males. PMID- 18427136 TI - Risk factors for aborted cardiac arrest and sudden cardiac death in children with the congenital long-QT syndrome. AB - BACKGROUND: The congenital long-QT syndrome (LQTS) is an important cause of sudden cardiac death in children without structural heart disease. However, specific risk factors for life-threatening cardiac events in children with this genetic disorder have not been identified. METHODS AND RESULTS: Cox proportional hazards regression modeling was used to identify risk factors for aborted cardiac arrest or sudden cardiac death in 3015 LQTS children from the International LQTS Registry who were followed up from 1 through 12 years of age. The cumulative probability of the combined end point was significantly higher in boys (5%) than in girls (1%; P<0.001). Risk factors for cardiac arrest or sudden cardiac death during childhood included corrected QT interval [QTc] duration > 500 ms (hazard ratio [HR]; 2.72; 95% confidence interval [CI], 1.50 to 4.92; P=0.001) and prior syncope (recent syncope [< 2 years]: HR, 6.16; 95% CI 3.41 to 11.15; P<0.001; remote syncope [> or = 2 years]: HR, 2.67; 95% CI, 1.22 to 5.85; P=0.01) in boys, whereas prior syncope was the only significant risk factor among girls (recent syncope: HR, 27.82; 95% CI, 9.72 to 79.60; P<0.001; remote syncope: HR, 12.04; 95% CI, 3.79 to 38.26; P<0.001). Beta-blocker therapy was associated with a significant 53% reduction in the risk of cardiac arrest or sudden cardiac death (P=0.01). CONCLUSIONS: LQTS boys experience a significantly higher rate of fatal or near-fatal cardiac events than girls during childhood. A QTc duration > 500 ms and a history of prior syncope identify risk in boys, whereas prior syncope is the only significant risk factor among girls. Beta-blocker therapy is associated with a significant reduction in the risk of life-threatening cardiac events during childhood. PMID- 18427138 TI - Images in cardiovascular medicine. Persistent thebesian sinusoids presenting as ischemic heart disease. PMID- 18427137 TI - Stromal cell-derived factor-1alpha is cardioprotective after myocardial infarction. AB - BACKGROUND: Heart disease is a leading cause of mortality throughout the world. Tissue damage from vascular occlusive events results in the replacement of contractile myocardium by nonfunctional scar tissue. The potential of new technologies to regenerate damaged myocardium is significant, although cell-based therapies must overcome several technical barriers. One possible cell-independent alternative is the direct administration of small proteins to damaged myocardium. METHODS AND RESULTS: Here we show that the secreted signaling protein stromal cell-derived factor-1alpha (SDF-1alpha), which activates the cell-survival factor protein kinase B (PKB/Akt) via the G protein-coupled receptor CXCR4, protected tissue after an acute ischemic event in mice and activated Akt within endothelial cells and myocytes of the heart. Significantly better cardiac function than in control mice was evident as early as 24 hours after infarction as well as at 3, 14, and 28 days after infarction. Prolonged survival of hypoxic myocardium was followed by an increase in levels of vascular endothelial growth factor protein and neoangiogenesis. Consistent with improved cardiac function, mice exposed to SDF-1alpha demonstrated significantly decreased scar formation than control mice. CONCLUSIONS: These findings suggest that SDF-1alpha may serve a tissue-protective and regenerative role for solid organs suffering a hypoxic insult. PMID- 18427139 TI - Letter by Hibbert et al regarding article, "Smooth muscle cells healing atherosclerotic plaque disruptions are of local, not blood, origin in apolipoprotein E knockout mice". PMID- 18427141 TI - Diastolic dysfunction: one piece of the heart failure with normal ejection fraction puzzle. PMID- 18427142 TI - Studies of drug-eluting stents: to each his own? PMID- 18427143 TI - Cardiac allograft vasculopathy: recent developments. AB - Cardiac allograft vasculopathy (CAV) continues to limit the long-term success of cardiac transplantation. Recent insights have underscored the fact that innate and adaptive immune responses are involved in the pathogenesis of CAV. Vascular lesions are the result of cumulative endothelial injuries induced both by alloimmune responses and by nonspecific insults (including ischemia-reperfusion injury, viral infections, and metabolic disorders) in the context of impaired repair mechanisms. Intravascular ultrasound is the most sensitive method for detection of CAV, and progressive intimal thickening in the first posttransplant year identifies patients at high risk for future cardiovascular events. Encouraging results with regard to the detection of CAV by noninvasive methods should be an incentive to apply routine noninvasive imaging during mid- to long term follow-up. Improved immunosuppressive drugs, including mycophenolate mofetil and proliferation signal inhibitors, as well as statins (in part via immunomodulation), have beneficial effects on CAV progression, although there is still a need to confirm the impact of vasodilators in improving outcome after heart transplantation. Coronary revascularization for CAV is only palliative, with no long-term survival benefit. Three main strategies for CAV prevention are currently under investigation: inhibition of growth factors and cytokines, cell therapy, and tolerance induction. However, because individual responses to an allograft change over time, assays to monitor the recipient's immune response and individualized methods for therapeutic immune modulation are clearly needed. PMID- 18427144 TI - Antioxidant gene therapy for cardiovascular disease: current status and future perspectives. AB - Excessive production of reactive oxygen species has been implicated to play an important role in a number of cardiovascular pathologies, including hypertension, atherosclerosis, myocardial infarction, ischemia/reperfusion injury, and restenosis after angioplasty or venous bypass grafting. The formation of reactive oxygen species is balanced out by antioxidant defenses, and augmenting this defense by antioxidant therapies could therefore provide a potential means to treat conditions in which the formation of reactive oxygen species exceeds the capability of natural protective mechanisms. In this review, we summarize the studies in which antioxidant gene therapy has been used successfully to treat cardiovascular diseases. We also discuss the current limitations of antioxidant gene therapy and envision future therapeutic targets and methodological approaches for an improved outcome. PMID- 18427145 TI - Therapeutic uses of inorganic nitrite and nitrate: from the past to the future. AB - Potential carcinogenic effects, blue baby syndrome, and occasional intoxications caused by nitrite, as well as the suspected health risks related to fertilizer overuse, contributed to the negative image that inorganic nitrite and nitrate have had for decades. Recent experimental studies related to the molecular interaction between nitrite and heme proteins in blood and tissues, the potential role of nitrite in hypoxic vasodilatation, and an unexpected protective action of nitrite against ischemia/reperfusion injury, however, paint a different picture and have led to a renewed interest in the physiological and pharmacological properties of nitrite and nitrate. The range of effects reported suggests that these simple oxyanions of nitrogen have a much richer profile of biological actions than hitherto assumed, and several efforts are currently underway to investigate possible beneficial effects in the clinical arena. We provide here a brief historical account of the medical uses of nitrite and nitrate over the centuries that may serve as a basis for a careful reassessment of the health implications of their exposure and intake and may inform investigations into their therapeutic potential in the future. PMID- 18427146 TI - Images in cardiovascular medicine. Cardiac resynchronization by restoration of native ventricular activation: reversal of iatrogenic mitral regurgitation and heart failure. PMID- 18427148 TI - Individualizing treatment of patients with myeloma in the era of novel agents. AB - Progress in the understanding of multiple myeloma (MM) cell biology has led to the identification of new relevant prognostic factors and subsequently different risk groups. This concept, together with the recent discovery of new drugs with novel mechanisms of action, will probably lead to individualized treatment according to the different patients' characteristics. In this review, we focus on current available agents already approved for MM, and discuss individualized treatment approaches for both transplantation candidates (subdivided into standard and high-risk patients) and elderly patients. Future progress in MM will be based on using science to inform the design of the optimal combined treatments, and high throughput assays that can assess the ability of combination therapies to induce death of MM cells, both alone and in the bone marrow microenvironment. PMID- 18427147 TI - Risedronate prevents bone loss in breast cancer survivors: a 2-year, randomized, double-blind, placebo-controlled clinical trial. AB - PURPOSE: Limited data are available on the efficacy of oral bisphosphonate therapy in breast cancer survivors. Our goal was to examine prevention of breast cancer-related bone loss in this cohort. PATIENTS AND METHODS: Eighty-seven postmenopausal women after chemotherapy for breast cancer were randomly assigned to once-weekly risedronate 35 mg or placebo for 24 months. Outcomes included bone mineral density (BMD) and turnover markers. RESULTS: At study initiation, 13% of patients were on an aromatase inhibitor (AI). After 24 months, there were differences of 1.6 to 2.5% (P < .05) at the spine and hip BMD between the placebo and risedronate groups. At study completion, 44% were on an AI. Adjusting for an AI, women on placebo plus AI had a decrease in BMD of (mean +/- SE) 4.8% +/- 0.8% at the spine and 2.8% +/- 0.5% at the total hip (both P < .001). In women on risedronate + AI, the spine decreased by 2.4% +/- 1.1% (P < .05) and was stable at the hip. Women in the placebo group not on an AI, maintained BMD at the spine, and had a 1.2% +/- 0.5% loss at the total hip (P < .05). Women who received risedronate but no AI had the greatest improvement in BMD of 2.2% +/- 0.9% (P < .05) at the total hip. Bone turnover was reduced with risedronate. There were no differences in adverse events between the groups. CONCLUSION: We conclude that in postmenopausal women with breast cancer with or without AI therapy, once-weekly oral risedronate was beneficial for spine and hip BMD, reduced bone turnover, and was well tolerated. PMID- 18427149 TI - HIV-specific differences in outcome of squamous cell carcinoma of the anal canal: a multicentric cohort study of HIV-positive patients receiving highly active antiretroviral therapy. AB - PURPOSE: To define clinical outcome after definitive chemoradiotherapy (CRT) of anal carcinoma in HIV-infected patients treated with highly active antiretroviral therapy (HAART). PATIENTS AND METHODS: A multicentric cohort comparison of 40 HIV positive patients with HAART and 81 HIV-negative patients treated with radiotherapy (RT) or CRT was retrospectively performed. Local disease control (LC), relapse-free survival (RFS), overall survival (OS), cancer-specific survival (CSS), toxicity, and prognostic factors were investigated. RESULTS: HIV positive patients were younger (mean age, 48 v 62 years; P < .0005), predominantly male (93% v 25%; P < .0005), and with early-stage (P = .06) and large-cell histology (90% v 67%; P = .005) disease. RT or CRT resulted in complete response in 92% (HIV positive) and 96% (HIV negative) of cases. Five year OS was 61% (95% CI, 44% to 78%) in HIV-positive and 65% (95% CI, 53% to 77%) in HIV-negative patients (median follow-up, 36 months). Five-year LC was 38% (95% CI, 5% to 71%) in HIV-positive and 87% (95% CI, 79% to 95%) in HIV-negative patients (P = .008) compromising CSS and sphincter preservation. Grade 3/4 acute skin (35% v 17% [HIV negative]; P = .04) and hematologic (33% v 12% [HIV negative]; P = .08) toxicity together approximated 50% in HIV-positive patients. RFS in HIV-positive patients was associated with RT dose (P = .08) and severe acute skin toxicity (P = .04). CONCLUSION: Long-term LC and acute toxicity represent major clinical challenges in HIV-positive patients with anal carcinoma. Even if fluoropyrimidine-based CRT is feasible and may result in similar response rates and OS as in HIV-negative patients, improved treatment strategies with better long-term outcome are warranted. PMID- 18427150 TI - Higher doses of lenalidomide are associated with unacceptable toxicity including life-threatening tumor flare in patients with chronic lymphocytic leukemia. AB - PURPOSE: Lenalidomide is a novel therapeutic agent with uncertain mechanism of action that is clinically active in myelodysplastic syndrome (MDS) and multiple myeloma (MM). Application of high (MM) and low (MDS) doses of lenalidomide has been reported to have clinical activity in CLL. Herein, we highlight life threatening tumor flare when higher doses of lenalidomide are administered to patients with CLL and provide a potential mechanism for its occurrence. PATIENTS AND METHODS: Four patients with relapsed CLL were treated with lenalidomide (25 mg/d for 21 days of a 28-day cycle). Serious adverse events including tumor flare and tumor lysis are summarized. In vitro studies examining drug-induced apoptosis and activation of CLL cells were also performed. RESULTS: Four consecutive patients were treated with lenalidomide; all had serious adverse events. Tumor flare was observed in three patients and was characterized by dramatic and painful lymph node enlargement resulting in hospitalization of two patients, with one fatal outcome. Another patient developed sepsis and renal failure. In vitro studies demonstrated lenalidomide-induced B-cell activation (upregulation of CD40 and CD86) corresponding to degree of tumor flare, possibly explaining the tumor flare observation. CONCLUSION: Lenalidomide administered at 25 mg/d in relapsed CLL is associated with unacceptable toxicity; the rapid onset and adverse clinical effects of tumor flare represent a significant limitation of lenalidomide use in CLL at this dose. Drug-associated B-cell activation may contribute to this adverse event. Future studies with lenalidomide in CLL should focus on understanding this toxicity, investigating patients at risk, and investigating alternative safer dosing schedules. PMID- 18427151 TI - Granulocyte-macrophage colony-stimulating factor potentiates rituximab in patients with relapsed follicular lymphoma: results of a phase II study. AB - PURPOSE: We hypothesized that granulocyte-macrophage colony-stimulating factor (GM-CSF) could potentiate the clinical activity of rituximab given its individual and cooperative effects on Fc gamma RIIa- and Fc gamma RIIIa-expressing cells. A phase II clinical study combining GM-CSF and rituximab was initiated in patients with relapsed follicular lymphoma (FL) to determine the clinical and biologic responses, as well as safety of the combination. PATIENTS AND METHODS: Thirty three patients with relapsed FL were treated with GM-CSF 5 microg/kg/d on days 1 to 8 and rituximab 375 mg/m(2) on day 5 of each 21-day cycle for four cycles. Clinical response and tolerability were examined according to international criteria. Biologic monitoring included evaluation of immune cells involved in rituximab activity. RESULTS: Of 33 evaluated patients, a 70% overall response rate (complete response plus complete response unconfirmed, 45%) and a median progression-free survival (PFS) of 16.5 months were achieved. Outcome was influenced by the quality of response and the Follicular Lymphoma International Prognostic Index (FLIPI), where low- and intermediate-risk FLIPI groups were associated with significantly better PFS. After treatment there was a significant increase in granulocyte and monocyte counts. Examination of dendritic cell response showed an overall increase in plasmacytoid dendritic cells, especially in non-complete response patients, after treatment. Addition of GM-CSF did not impair tolerance to rituximab, and adverse events were rare and mild. DISCUSSION: GM-CSF plus rituximab results in high response rates, along with a tolerable safety profile in patients with relapsed or progressive FL. The improved efficacy over rituximab monotherapy may be due to increases seen in monocyte, granulocyte, and dendritic cell populations. PMID- 18427152 TI - Mammography before diagnosis among women age 80 years and older with breast cancer. AB - PURPOSE: Screening mammography guidelines for patients age 80 years and older are variable. We determined the effect of mammography use on stage at breast cancer diagnosis and survival among women of this age range. PATIENTS AND METHODS: We used the linked Surveillance, Epidemiology, and End Results-Medicare database to evaluate 12,358 women >or= 80 years of age diagnosed with breast cancer between 1996 and 2002. Patients were grouped according to number of mammograms during the 60 months before diagnosis: nonusers (0 mammograms), irregular users (one to two mammograms), and regular users (three or more mammograms). Effects of mammography on disease stage (I to IIa v IIb to IV) and survival were determined by logistic regression and Cox proportional hazards analyses. RESULTS: Percentages of women with nonuse, irregular use, and regular use of mammography during the 5 years preceding diagnosis were 49%, 29%, and 22%, respectively. On multivariate analysis, patients were 0.37 times less likely to present with late-stage cancer for each mammogram obtained (odds ratio, 0.63; 95% CI, 0.63 to 0.67). Breast cancer-specific 5-year survival among nonusers was 82%, that among irregular users was 88%, and that among regular users was 94%. However, survival from causes other than breast cancer was also associated with mammography use, suggesting a bias for healthier patients to undergo mammography. CONCLUSION: Regular mammography among women >or= 80 years of age was associated with earlier disease stage, although improved survival remains difficult to demonstrate. Health care providers should consider discussing the potential benefits of screening mammography with their older patients, particularly for those without significant comorbidity. PMID- 18427153 TI - GPR119: "double-dipping" for better glycemic control. PMID- 18427154 TI - Insulin-like growth factor binding protein-2: a novel regulator of skeletal gender differences? PMID- 18427156 TI - Lipid composition of microdomains is altered in a cell model of Gaucher disease. AB - The formation of cholesterol and sphingolipids into specialized liquid-ordered membrane microdomains (rafts) has been proposed to function in the intracellular sorting and transport of proteins and lipids. Defined by biochemical criteria, rafts resist solubilization in nonionic detergents, enabling them to be isolated as detergent-resistant membranes (DRM). In this study, we characterized the lipid composition of DRM from a cell model of the sphingolipid storage disorder, Gaucher disease, in which the catabolism of the sphingolipid glucosylceramide (GC) is impaired. In this cell model, we showed that GC accumulated primarily in the DRM, with smaller secondary increases in ceramide, dihexosylceramide, trihexosylceramide, and phosphatidylglycerol. This suggested that not only was lipid metabolism altered as a consequence of the cells' inability to degrade GC, but this affected the DRM rather than other regions of the membrane. This increase in lipids in the DRM may be responsible for the altered lipid and protein sorting seen in Gaucher disease. Analysis of individual lipid species revealed preservation of the shorter and fully saturated fatty acid species in the DRM, suggesting that the highly ordered and tightly packed nature of the DRM is maintained. PMID- 18427157 TI - Human Reproduction. Editor's choice. PMID- 18427158 TI - Robust electrocardiogram (ECG) beat classification using discrete wavelet transform. AB - This paper presents a robust technique for the classification of six types of heartbeats through an electrocardiogram (ECG). Features extracted from the QRS complex of the ECG using a wavelet transform along with the instantaneous RR interval are used for beat classification. The wavelet transform utilized for feature extraction in this paper can also be employed for QRS delineation, leading to reduction in overall system complexity as no separate feature extraction stage would be required in the practical implementation of the system. Only 11 features are used for beat classification with the classification accuracy of approximately 99.5% through a KNN classifier. Another main advantage of this method is its robustness to noise, which is illustrated in this paper through experimental results. Furthermore, principal component analysis (PCA) has been used for feature reduction, which reduces the number of features from 11 to 6 while retaining the high beat classification accuracy. Due to reduction in computational complexity (using six features, the time required is approximately 4 ms per beat), a simple classifier and noise robustness (at 10 dB signal-to noise ratio, accuracy is 95%), this method offers substantial advantages over previous techniques for implementation in a practical ECG analyzer. PMID- 18427159 TI - Minimally obtrusive wearable device for continuous interactive cognitive and neurological assessment. AB - This study demonstrates the feasibility of a minimally obtrusive wearable system that can assess cognitive performance continuously throughout normal life activities by excitation of the peripheral nervous system and detection of the central nervous system response. The new concept was tested with one possible implementation as a device the size of a wristwatch which interrogates the subject by means of haptic excitation (vibration) and records the responses (subtle hand movements detected by accelerometers). The system was programmed to perform simple reaction time trials and was tested with ten volunteers during 8 h of their normal daytime activities. Results indicate that the volunteers responded properly to most of the interrogations (>95%) and that the impact of the device on everyday activities was not significant. The ability to assess cognitive capabilities of individuals continuously during everyday activities could have far-reaching implications for diagnostics and treatment of many different neurological conditions. PMID- 18427160 TI - Gastrointestinal arrhythmias are associated with statistically significant fluctuations in systemic information dimension. AB - Although cardiac arrhythmias have been studied extensively, little is known about arrhythmic phenomena in the gastrointestinal (GI) system. In this study, we demonstrate for the first time that the onset of GI arrhythmias is associated with statistically significant fluctuations in the information dimension of the associated systems. We induced gastric and intestinal arrhythmias in pigs using surgical stomach division and mesenteric artery ligation, respectively. Both conditions lead to a decreased supply of blood to the GI tract, which is associated in humans with various potentially lethal conditions including chronic mesenteric ischemia, whose mortality rate is over 60%. During our experiments, we recorded simultaneous magnetocardiographic, magnetogastrographic and magnetoenterographic signals and concluded that, when GI circulation is compromised, the information dimensionality of the system fluctuates significantly. In conclusion, dimensionality may be an important diagnostic factor for the characterization of arrhythmias in the context of GI pathophysiology. PMID- 18427161 TI - Non-invasive monitoring of chewing and swallowing for objective quantification of ingestive behavior. AB - A methodology of studying of ingestive behavior by non-invasive monitoring of swallowing (deglutition) and chewing (mastication) has been developed. The target application for the developed methodology is to study the behavioral patterns of food consumption and producing volumetric and weight estimates of energy intake. Monitoring is non-invasive based on detecting swallowing by a sound sensor located over laryngopharynx or by a bone-conduction microphone and detecting chewing through a below-the-ear strain sensor. Proposed sensors may be implemented in a wearable monitoring device, thus enabling monitoring of ingestive behavior in free-living individuals. In this paper, the goals in the development of this methodology are two-fold. First, a system comprising sensors, related hardware and software for multi-modal data capture is designed for data collection in a controlled environment. Second, a protocol is developed for manual scoring of chewing and swallowing for use as a gold standard. The multi modal data capture was tested by measuring chewing and swallowing in 21 volunteers during periods of food intake and quiet sitting (no food intake). Video footage and sensor signals were manually scored by trained raters. Inter rater reliability study for three raters conducted on the sample set of five subjects resulted in high average intra-class correlation coefficients of 0.996 for bites, 0.988 for chews and 0.98 for swallows. The collected sensor signals and the resulting manual scores will be used in future research as a gold standard for further assessment of sensor design, development of automatic pattern recognition routines and study of the relationship between swallowing/chewing and ingestive behavior. PMID- 18427162 TI - The ABA effect on the accumulation of an invertase inhibitor transcript that is driven by the CAMV35S promoter in ARABIDOPSIS. AB - Invertase (beta-D-fructofuranosidase; EC 3.2.1.26) catalyzes the conversion of sucrose into glucose and fructose and is involved in an array of important processes, including phloem unloading, carbon partitioning, the response to pathogens, and the control of cell differentiation and development. Its importance may have caused the invertases to evolve into a multigene family whose members are regulated by a variety of different mechanisms, such as pH, sucrose levels, and inhibitor proteins. Although putative invertase inhibitors in the Arabidopsis genome are easy to locate, few studies have been conducted to elucidate their individual functions in vivo in plant growth and development because of their high redundancy. In this study we assessed the functional role of the putative invertase inhibitors in Arabidopsis by generating transgenic plants harboring a putative invertase inhibitor gene under the control of the CaMV35S promoter. A transgenic plant that expressed high levels of the putative invertase inhibitor transcript when grown under normal conditions was chosen for the current study. To our surprise, the stability of the invertase inhibitor transcripts was shown to be down-regulated by the phytohormone ABA (abscisic acid). It is well established that ABA enhances invertase activity in vivo but the underlying mechanisms are still poorly understood. Our results thus suggest that one way ABA regulates invertase activity is by down-regulating its inhibitor. PMID- 18427163 TI - Th17 cells and rheumatoid arthritis--from the standpoint of osteoclast differentiation--. AB - Rheumatoid arthritis is a chronic disease that affects multiple joints. It is considered to be an autoimmune disease in which a T helper (Th)-1 type response has been implicated to play an important pathogenetic role. As osteoclasts, cells that resorb bone, play a crucial part in the bone destruction that occurs in RA, we and others have investigated the pathophysiology of these cells. The findings that interferon (IFN)-gamma strongly inhibits osteoclastogenesis and that interleukin (IL)-17 has the ability to enhance osteoclast differentiation have cast doubt on the hypothesis that RA is a Th1 disease. In this review, I describe the relationship between Th cells, the so-called "commander" of the immune response, and RA, mainly from the viewpoint of the environments Th cells create for the excessive differentiation and function of osteoclasts, resulting in the destruction of bone. PMID- 18427164 TI - Th17 Cells and autoimmune encephalomyelitis (EAE/MS). AB - Multiple sclerosis (MS) is a CD4+ T cell-mediated autoimmune disease affecting the central nervous system. It was largely accepted that Th1 cells driven by IL 12 were pathogenic T cells in human MS and experimental autoimmune encephalomyelitis, an animal model of MS. Recent data have established that IL-17 producing CD4+ T cells, driven by IL-23 and referred to as Th17 cells, play a pivotal role in the pathogenesis of EAE. A combination of TGF-beta and IL-6 induce Th17 cell lineage commitment via expression of transcription factor RORgammat. Th17 cells and induced Foxp3+ T regulatory cells are in reciprocal position in the T cell lineage commitment governed by TGF-beta and IL-6. The vitamin A metabolite retinoic acid is involved in this process via TGF-beta dependent induction of Foxp3. We have demonstrated that human Th17 cells could be identified as CCR2+ CCR5- memory CD4+ T cells. It is becoming clear that IL 23/Th17 axis also plays an important role in the pathogenesis of various human autoimmune diseases including MS. Additionally, accumulating evidences raise a possibility that CCR2 on Th17 cells may be a therapeutic target in MS. PMID- 18427165 TI - Th17 and allergy. AB - The identification of novel helper T (Th) cell subsets, i.e., IL-17-producing Th cells (Th17 cells) and regulatory T cells (Treg cells), provided new insight into our understanding of the molecular mechanisms involved in the development of infectious and autoimmune diseases as well as immune responses, and thus led to revision of the classic Th1/Th2 paradigm. Several current lines of evidence from gene-deficient mice indicate that IL-17 and Th17 cells, but not IFN-gamma and Th1 cells, are responsible for the development of autoimmune diseases such as murine arthritis and encephalomyelitis, which have classically been considered to be Th1 mediated disorders. Th17 cells may also contribute to the pathogenesis of classically recognized Th2-mediated allergic disorders. In this review, we summarize the current knowledge regarding IL-17 and Th17 cells and discuss their potential roles in the pathogenesis of allergic disorders. PMID- 18427166 TI - Human Th17 cell clones and natural immune responses. AB - Immunomodulators such as lipopolysaccharides (LPS) and forskolin change the nature of dendritic cells (DCs) to induce Th1 and Th2 cells, respectively, thereby designated Th1 or Th2 adjuvants. Recent studies showed that Th17-inducing activity can be carried by certain polysaccharides such as beta-glucan derived from Candia albicans. Such activities can be scrutinized by using MLR, cAMP and possibly, differential expression of Notch ligand isoforms. In this review article, we also introduce an effective method to establish human Th17 cell clones and a transcriptome analysis using human Th subpopulations. In vivo relevance to human Th17 responses is discussed. PMID- 18427167 TI - A 13-year study of Japanese cedar pollinosis in Japanese schoolchildren. AB - BACKGROUND: Japanese cedar pollen (JCP) sensitization and Japanese cedar pollinosis (JCPS) appear to be increasingly prevalent in younger children. The present study investigated factors affecting JCP sensitization and JCPS development in school children. METHODS: In May or June each year from 1994 to 2006, 275-510 children were assessed for serum JCP-IgE and house dust mite (HDM) IgE levels, and surveyed regarding rhinoconjunctival symptoms. RESULTS: Strong JCP sensitization (IgE > or = 17.5 U(A)/ml) was associated with age (odds ratio (OR) = 2.65), the amount of dispersed pollen in the observed year (OR = 2.03) and in the year following birth (OR = 1.51), the month of birth (OR = 2.18), and the recent birth cohort (OR = 1.96). Symptoms were negatively correlated with the recent birth cohort (OR = 0.69) after adjusting for JCP-IgE levels. Strong HDM sensitization was associated with gender (OR = 0.65 for girls) and the recent birth cohort (OR = 1.76). CONCLUSIONS: JCP sensitization appeared to be associated with the recent birth cohort and to increases in dispersed pollen just after birth and in the observed season. Although the recent birth cohort was more easily sensitized, they were not more likely to develop symptoms. In contrast to JCP sensitization, strong HDM sensitization appeared to develop prior to commencement of primary school and was more likely to affect boys. PMID- 18427168 TI - Does drug-induced hypersensitivity syndrome elicit bullous pemphigoid? PMID- 18427198 TI - Human leukocyte antigen-DQ alleles and haplotypes and their associations with resistance and susceptibility to HIV-1 infection. AB - OBJECTIVES: To determine the association of DQ antigens with resistance and susceptibility to HIV-1. DESIGN: Despite repeated exposure to HIV-1, a subset of women in the Pumwani Sex Worker cohort established in Nairobi, Kenya in 1985 have remained HIV-1 negative for at least 3 years and are classified as resistant. Differential susceptibility to HIV-1 infection is associated with HIV-1 specific CD4 and CD8 T cell responses. As human leukocyte antigen-DQ antigens present viral peptides to CD4 cells, we genotyped human leukocyte antigen -DQ alleles for 978 women enrolled in the cohort and performed cross-sectional and longitudinal analyses to identify associations of human leukocyte antigen -DQ with resistance/susceptibility to HIV-1. METHODS: DQA1 and DQB1 were genotyped using taxonomy-based sequence analysis. SPSS 13.0 was used to determine associations of DQ alleles/haplotypes with HIV-1 resistance, susceptibility, and seroconversion rates. RESULTS: Several DQB1 alleles and DQ haplotypes were associated with resistance to HIV-1 infection. These included DQB1*050301 (P = 0.055, Odds Ratio = 12.77, 95% Confidence Interval = 1.44-112), DQB1*0603 and DQB1*0609 (P = 0.037, Odds Ratio = 3.25, 95% Confidence Interval = 1.12-9.47), and DQA1*010201 DQB1*0603 (P = 0.044, Odds Ratio = 17.33, 95% Confidence Interval = 1.79-168). Conversely, DQB1*0602 (P = 0.048, Odds Ratio = 0.68, 95% Confidence Interval = 0.44-1.05) and DQA1*010201-DQB1*0602 (P = 0.039, Odds Ratio = 0.64, 95% Confidence Interval = 0.41-1.03) were overrepresented in the HIV-1 infected population. DQA1*0504-DQB1*0201, DQA1*010201-DQB1*0201, DQA1*0402-DQB1*0402 and DQA1*0402-DQB1*030101 genotypes were only found in HIV-1 positive subjects (Odds Ratio = 0.30-0.31, 95% Confidence Interval = 0.03-3.70), and these women seroconverted rapidly. The associations of these DQ alleles and haplotypes with resistance and susceptibility to HIV-1 were independent of the previously reported human leukocyte antigen-DRB*01, human leukocyte antigen A2/6802, and human leukocyte antigen-A*2301. CONCLUSION: The associations of DQ alleles and haplotypes with resistance and susceptibility to HIV-1 emphasize the importance of human leukocyte antigen-DQ and CD4 in anti-HIV-1 immunity. PMID- 18427197 TI - Children and HIV/AIDS: from research to policy and action in resource-limited settings. PMID- 18427199 TI - Genetic variation in mother-child acute seroconverter pairs from Zambia. AB - OBJECTIVE: To characterize the envelope (Env) glycoprotein of HIV-1 in mother infant pairs (MIP) that underwent near simultaneous or acute-phase seroconversion, we examined the Env sequence of the transmitted viruses and compare viral evolution within the pair. DESIGN: Three MIP from a Zambian cohort that seroconverted at the same sampling time were identified and followed longitudinally. METHODS: The V1-V5 region of the HIV-1 Env gene was sequenced for each sample collected. Phylogenetic and population genetics analyses were carried out to subtype the viruses, estimate relationships among viral genotypes, and compare molecular evolution between the viral populations. RESULTS: Genetic analyses demonstrated a close intrapair relationship between viral sequences from each MIP. Transmission involved several closely related viral genotypes and did not result in a reduction in viral diversity. Amino acid changes were not evenly distributed along Env V1-V5 but concentrated in concordant areas within each MIP. Several positions under positive selection were shared between the MIP viruses. Interestingly, selective pressure on the virus was higher in the infants than in the mothers. CONCLUSIONS: In contrast to most cases of perinatal transmission of HIV-1 from chronically infected mothers, there is no evidence of a genetic bottleneck in the transmitted viruses in these three instances of acute seroconversion. The longitudinal changes in the amino acids are in similar positions in Env for the MIP, suggesting shared evolutionary constrains among the closely related viruses infecting the MIP; such constrains may lead to similar genetic changes in the virus in two different hosts. PMID- 18427200 TI - Role of HIV and human herpesvirus-8 infection in pulmonary arterial hypertension. AB - BACKGROUND: Previous work has found a high prevalence of pulmonary arterial hypertension in HIV-infected persons, but establishment of a causal relationship has been limited by the lack of well characterized contemporaneous HIV-uninfected comparator groups. Among HIV-uninfected persons, human herpesvirus-8 (HHV-8) has also been linked to pulmonary arterial hypertension (PAH), but whether this relationship occurs among HIV-infected persons - who have among the highest prevalence of HHV-8 infection - has not been examined. METHODS AND RESULTS: We echocardiographically calculated pulmonary artery systolic pressure and measured HHV-8 antibodies in HIV-infected and HIV-uninfected adults. Among the 196 HIV infected participants, the median pulmonary artery systolic pressure (PASP) was 27.5 mmHg and 35.2% had PASP greater than 30 mmHg. This compared to a median of 22 mmHg among 52 HIV-uninfected participants in whom 7.7% had a PASP greater than 30 mmHg (P < 0.001). After adjustment for injecting drug and stimulant use, smoking, age, and gender, HIV-infected participants had 5.1 mmHg higher mean PASP and seven fold greater odds of having a PASP greater than 30 mmHg (P < 0.001). Although we found no association between HHV-8 and PAH among all HIV-infected participants, a borderline relationship was present when restricting to those without risk factors for PAH. CONCLUSION: HIV-infected persons have a high prevalence of elevated PASP, which is independent of other risk factors for PAH. This suggests a causal role of HIV in PAH and emphasizes the need to understand the natural history of PAH in this setting. A role for HHV-8 infection in PAH remains much less definitive. PMID- 18427201 TI - Comparison of algorithms that interpret genotypic HIV-1 drug resistance to determine the prevalence of transmitted drug resistance. AB - OBJECTIVE: We compared eight genotypic interpretation methods to determine whether the method used would affect the rates of reported transmitted drug resistance. DESIGN: Retrospective cohort study. METHODS: For the International AIDS Society-USA method we classified a mutation as resistant if it was a 'major' resistance-associated mutation. For the Stanford algorithm, we classified a mutation as resistant if the score was at least 60 (Stanford 60), and alternatively, if the score was at least 30 (Stanford 30). For Agence Nationale de Recherches sur le SIDA and Rega, we interpreted resistance as either 'intermediate resistance' or 'resistance' (ANRS 1 and Rega 1), and 'resistance' only (ANRS 2 and Rega 2). We also used the calibrated population resistance algorithm. We then determined the rates of transmitted drug resistance within the Acute Infection Early Disease Research Program cohort (n = 1311) enrolled between March 1995 and August 2006 using each method; agreement was assessed using kappa coefficients. RESULTS: Differences in estimated rates of transmitted drug resistance using International AIDS Society-USA, calibrated population resistance, Stanford 30, ANRS 1, Rega 1 and Rega 2 methods were mostly minor for resistance to protease and non-nucleoside reverse transcriptase inhibitors (1% range) and more pronounced for nucleoside reverse transcriptase inhibitors (5% range). For these methods kappa agreement was substantial or almost perfect across all drug classes. The Stanford 60 was most conservative. CONCLUSIONS: The persistent high rates of transmitted drug resistance support the need for continued genotypic surveillance. The currently available interpretation algorithms can be used for this purpose. PMID- 18427202 TI - CD4+ count and risk of non-AIDS diseases following initial treatment for HIV infection. AB - BACKGROUND: Reductions in AIDS-related morbidity and mortality following the advent of combination antiretroviral therapy have coincided with relative increases in chronic non-AIDS end-organ diseases among HIV+ patients. OBJECTIVE: To examine the association of latest CD4+ counts with risk of non-AIDS diseases in a cohort of 1397 patients who initiate antiretroviral therapy. METHODS: CD4+ counts and HIV RNA levels along with fatal, and non-fatal, AIDS and non-AIDS diseases (liver, cardiovascular, renal, and cancer) were assessed over a median follow-up of 5 years. Cox proportional regression models were used to study risk associations. RESULTS: A total of 227 patients experienced an AIDS event and 80 patients developed a non-AIDS disease event. Both AIDS and non-AIDS diseases rates (events/100 person-years), respectively, declined with higher latest CD4+ counts: 13.8 and 2.1 with latest CD4+ counts less than 200 cells/microl; 2.0 and 1.7 for counts of 200-350 cells/microl; and 0.7 and 0.7 for counts greater than 350 cells/microl. After adjusting for baseline covariates and the latest HIV RNA level, risk of AIDS and non-AIDS diseases were lowered by 44% (95% confidence interval for hazard ratio 0.50-0.62, P < 0.01) and 14% (95% confidence interval for hazard ratio 0.77-0.96, P = 0.01), respectively, for each 100 cell/microl higher latest CD4+ count. CONCLUSION: Higher CD4+ counts on antiretroviral therapy are associated with lower rates of non-AIDS diseases and AIDS. These findings expand our understanding of the implications of HIV-related immunodeficiency and motivate randomized studies to evaluate the effects of antiretroviral therapy on a broad set of clinical outcomes at CD4+ counts greater than 350 cells/microl. PMID- 18427203 TI - Insulin resistance predicts endothelial dysfunction and cardiovascular risk in HIV-infected persons on long-term highly active antiretroviral therapy. AB - OBJECTIVE: Cardiovascular disease risk among persons with HIV is likely multifactorial, thus testing a variety of available noninvasive vascular ultrasound and other surrogate tests may yield differing results. To address this issue, we assessed multiple metabolic and clinical predictors of endothelial function and carotid intima-media thickness in HIV-infected subjects and compared results with HIV-negative controls. DESIGN: Prospective, cross-sectional study of 50 HIV-infected, healthy adults on stable highly active antiretroviral therapy matched to 50 HIV-negative controls by age, sex, race, and body mass index. METHODS: Flow-mediated vasodilation of the brachial artery, carotid intima-media thickness, dual energy X-ray absorptiometry (HIV-infected subjects), and fasting insulin, lipids, and oral glucose tolerance tests were performed. Results were compared between HIV-infected and control groups. RESULTS: Fifty percent of subjects were African-American with 34% women. Among HIV-infected, mean CD4 cell count was 547 cells/microl; 90% had HIV RNA less than 50 copies/ml. There were no significant differences between HIV-infected and control subjects with regard to brachial artery flow-mediated vasodilation or carotid intima-media thickness. In multivariate analyses of the HIV cohort, independent predictors of endothelial dysfunction (lower flow-mediated vasodilation) were increasing insulin resistance, greater alcohol consumption, and higher baseline brachial artery diameter (P < 0.05); predictors of increased carotid intima-media thickness were hypertension, higher trunk/limb fat ratio, and insulin resistance (P < 0.05). CONCLUSION: In this HIV cohort on modern highly active antiretroviral therapy with well controlled HIV, there were no significant differences with regard to preclinical markers of cardiovascular disease. Insulin resistance was a strong predictor of impaired brachial artery flow-mediated vasodilation and increased carotid intima-media thickness, and may be an important cardiovascular disease risk factor in the HIV population. PMID- 18427204 TI - Insulin resistance impairs rapid virologic response in HIV/hepatitis C virus coinfected patients on peginterferon-alfa-2a. AB - OBJECTIVES: To investigate the association between insulin resistance and rapid virologic response. DESIGN: All consecutive HIV/hepatitis C virus coinfected patients who started peg-interferon alpha-2a (180 microg/week) and ribavirin 1000 1200 mg/day were analysed. METHODS: Insulin resistance was defined according to the homeostasis model of assessment-insulin resistance calculated as fasting insulin (mIU/l) x fasting glucose (mmol/l)/22.5. Rapid virologic response was defined as testing negative for hepatitis C virus-RNA after 4 weeks of therapy. Fasting levels of insulin and glucose in plasma were measured in all patients on the first day of treatment. Hepatitis C virus-RNA was determined by quantitative PCR assay (version 3.0). Hepatitis C virus-RNA was measured by qualitative PCR assay (COBAS 2.0) after 4 weeks of treatment. RESULTS: Seventy-four HIV/hepatitis C virus coinfected patients were enrolled [mean age 41.7 years (SD 5.3), 61 men, 54.1% with advanced fibrosis (F3-4 according to METAVIR classification), 52.4% with infection by hepatitis C virus genotype 1 or 4]. Rapid virologic response was reached by 30 subjects. In the multivariate analysis the independent predictors of rapid virologic response were: genotype 1 or 4 [adjusted odds ratio 0.18 (0.06-0.55)], hepatitis C virus-RNA < 400.000 UI/ml [adjusted odds ratio 0.229 (0.09-0.92)] and homeostasis model of assessment-insulin resistance more than 3.00 [adjusted odds ratio 0.1 (0.05-0.6)]. CONCLUSION: The homeostasis model of assessment-insulin resistance score should be evaluated and possibly corrected before starting anti-hepatitis C virus therapy. PMID- 18427205 TI - The molecular epidemiology of HIV-1 envelope diversity during HIV-1 subtype C vertical transmission in Malawian mother-infant pairs. AB - OBJECTIVES: To study the relationship between HIV-1 subtype C genetic diversity and mother-to-child transmission and to determine if transmission of HIV-1 V1/V2 env variants occurs stochastically. DESIGN: Case-case-control study of Malawian mother-infant pairs consisting of 32 nontransmitting women, 25 intrauterine transmitters, and 23 intrapartum transmitters in Blantyre, Malawi. METHODS: A heteroduplex tracking assay against the highly variable HIV-1 env V1/V2 region was used to characterize the relationship between HIV-1 diversity and mother-to child transmission. The relative abundance of the maternal env variants was quantified and categorized as transmitted or nontransmitted based on the env variants detected in the infant plasma. The V1/V2 region was sequenced from two mother-infant pairs and a phylogenetic tree was built. RESULTS: No relationship was found between transmission and overall maternal env diversity. Infants had less diverse HIV-1 populations than their mothers, and intrauterine-infected infants had fewer V1/V2 variants and were more likely to harbor a homogeneous V1/V2 population than infants infected intrapartum. V1/V2 sequences cloned from two mother-infant transmission pairs support multiple env variant transmission when multiple variants are detected, rather than single variant transmission followed by diversification. Almost 50% of the HIV-infected infants contained V1/V2 env variants that were not detected in maternal plasma samples. Finally transmission of env variants was not related to their abundance in maternal blood. CONCLUSION: These data suggest that the predominant mechanism(s) of HIV-1 subtype C mother-to-child transmission differs by the timing of transmission and is unlikely to be explained by a simple stochastic model. PMID- 18427206 TI - Rapid scaling-up of antiretroviral therapy in 10,000 adults in Cote d'Ivoire: 2 year outcomes and determinants. AB - OBJECTIVE: To assess the rates and determinants of mortality, loss to follow-up and immunological failure in a nongovernmental organization-implemented program of access to antiretroviral treatment in Cote d'Ivoire. METHODS: In each new treatment center, professionals were trained in HIV care, and a computerized data system was implemented. Individual patient and program level determinants of survival, loss to follow-up and immunological failure were assessed by multivariate analysis. RESULTS: Between May 2004 and February 2007, 10,211 patients started antiretroviral treatment in 19 clinics (median preantiretroviral treatment CD4 cell count, 123 cells/microl; initial regimen zidovudine-lamivudine efavirenz, 20%; stavudine-lamivudine-efavirenz, 22%; stavudine-lamivudine nevirapine, 52%). At 18 months on antiretroviral treatment, the median gain in CD4 cell count was +202 cells/microl, the probability of death was 0.15 and the probability of being loss to follow-up was 0.21. In addition to the commonly reported determinants of impaired outcomes (low CD4 cell count, low BMI, low hemoglobin, advanced clinical stage, old age and poor adherence), two factors were also shown to independently jeopardize prognosis: male sex (men vs. women: hazard ratio = 1.52 for death, 1.27 for loss to follow-up, 1.31 for immunological failure); and attending a recently opened clinic (inexperienced vs. experienced centers: hazard ratio = 1.40 for death, 1.58 for loss to follow-up). None of the three outcomes was associated with the drug regimen. DISCUSSION: In this rapidly scaling-up program, survival and immune reconstitution were good; women and patients followed up in centers with longer experience had better outcomes; outcomes were similar in zidovudine/stavudine-based regimens and in efavirenz/nevirapine-based regimens. Decreasing the rate of loss to follow-up should now be the top priority in antiretroviral treatment rollout. PMID- 18427207 TI - Intervention to promote exclusive breast-feeding for the first 6 months of life in a high HIV prevalence area. AB - OBJECTIVES: We report on a nonrandomized intervention cohort study to increase exclusive breast-feeding rates for 6 months after delivery in HIV-positive and HIV-negative women in KwaZulu-Natal, South Africa. METHODS: Lay counselors visited women to support exclusive breast-feeding: four times antenatally, four times in the first 2 weeks postpartum and then fortnightly to 6 months. Daily feeding practices were collected at weekly intervals by separate field workers. Cumulative exclusive breast-feeding rates from birth were assessed by Kaplan Meier analysis and association with maternal and infant variables was quantified in a Cox regression analysis. FINDINGS: One thousand, two hundred and nineteen infants of HIV-negative and 1217 infants of HIV-positive women were followed postnatally. Median duration of exclusive breast-feeding was 177 (R = 1-180; interquartile range: 150-180) and 175 days (R = 1-180; interquartile range: 137 180) in HIV-negative and HIV-positive women, respectively. Using 24-h recall, exclusive breast-feeding rates at 3 and 5 months were 83.1 and 76.5%, respectively, in HIV-negative women and 72.5 and 66.7%, respectively, in HIV positive women. Using the most stringent cumulative data, 45% of HIV-negative and 40% of HIV-positive women adhered to exclusive breast-feeding for 6 months. Counseling visits were strongly associated with adherence to cumulative exclusive breast-feeding at 4 months, those who had received the scheduled number of visits were more than twice as likely to still be exclusively breast-feeding than those who had not (HIV-negative women: adjusted odds ratio: 2.07, 95% confidence interval: 1.56-2.74, P < 0.0001; HIV-positive women: adjusted odds ratio: 2.86, 95% CI 2.13-3.83, P < 0.0001). CONCLUSION: It is feasible to promote and sustain exclusive breast-feeding for 6 months in both HIV-positive and HIV-negative women, with home support from well trained lay counselors. PMID- 18427208 TI - The IL-6 system in HIV-1-infection and in HAART-related fat redistribution syndromes. AB - We determined the IL-6 -174 G>C single nucleotide polymorphism, IL-6 mRNA expression in subcutaneous adipose tissue (SAT) and IL-6 plasma levels in HIV-1 infected patients with and without lipodystrophy and uninfected controls. HIV-1 infected patients had a greater prevalence of the IL-6 -174 C/C genotype and the C allele, higher SAT IL-6 mRNA expression and plasma IL-6 levels than controls. The IL-6 -174 G>C genotype distribution and allele frequencies, SAT IL-6 mRNA expression and IL-6 plasma levels were non-significantly different between HIV-1 infected patients with and without lipodystrophy. PMID- 18427209 TI - Emergence of X4 usage among HIV-1 subtype C: evidence for an evolving epidemic in South Africa. AB - This study investigated the genotype and phenotype of HIV-1 isolates of 20 South African AIDS patients. We found the highest percentage of CXCR4 usage among primary isolates, in which 30% efficiently utilized CXCR4 and exhibited the syncytium-inducing phenotype. Phylogenetic analysis of env confirmed that 19 of the 20 were subtype C, and syncytium-inducing viruses had genetic changes in the V3 loop, characteristic of CXCR4 usage. Results imply that the frequency of CXCR4 utilizing subtype C is increasing with time. PMID- 18427210 TI - Similar hepatitis C virus RNA kinetics in HIV/hepatitis C virus monoinfected genotype 2 or 3 matched controls during hepatitis C virus combination therapy. AB - We prospectively studied early hepatitis C virus kinetics and sustained virological response rates in HIV/HCV coinfected (n = 13) and HCV monoinfected matched controls (n = 26) with HCV genotype 2/3 treated with pegylated interferon (peg-IFN) alpha-2a 135 microg/week plus ribavirin 11 mg/kg daily during 24 weeks. No significant difference in HCV-RNA decay was seen at any time point during the initial 12 weeks of therapy. Sustained virological response was achieved in 9/13 (69%) versus 20/26 (77%) patients (intent-to-treat), respectively. The lower-than standard peg-IFN dose offered high compliance and reasonable sustained virological response rates. PMID- 18427211 TI - Association of HIV infection and Mycobacterium ulcerans disease in Benin. AB - We investigated the association between Buruli ulcer and HIV by comparing the HIV 1/2 seroprevalence in a series of 426 Buruli ulcer patients and a sample of 613 residents of southern Benin. The overall HIV prevalence was 2.6% (11/426) among patients and 0.3% among controls (2/613), giving an odds ratio for the association between HIV and Buruli ulcer of 8.1 (95% confidence interval = 1.8 75; P = 0.003). PMID- 18427212 TI - Multifocal squamous cell carcinomas in an HIV-infected patient with a long-term voriconazole therapy. PMID- 18427213 TI - Antiretroviral therapy adherence in children: outcomes from Africa. PMID- 18427214 TI - Vaginal microbicides: the importance of effective distribution, retention and coating of the mucosa. PMID- 18427215 TI - Hepatitis E and jaundice in an HIV-positive pregnant woman. PMID- 18427216 TI - A low antiretroviral activity of the antihepatitis B drug entecavir may be enough to select for M184V in HIV-1. PMID- 18427217 TI - Immune reconstitution syndrome is not a single entity. PMID- 18427218 TI - Risk factors for falls during inpatient rehabilitation. AB - OBJECTIVE: To determine risk factors for falls during inpatient rehabilitation on the basis of admission data, and to assess the predictive value of the FIM instrument. DESIGN: One thousand four hundred seventy-two patients consecutively admitted to a large tertiary care rehabilitation center during 18 mos were included in this retrospective study. Events surrounding falls were reported by clinical staff. Demographic data, prehospital socioeconomic status, medical condition at admission, and admission FIM scores were analyzed using log-logistic regression model for their association with falls. RESULTS: One hundred forty (9.5%) patients fell at least once. Most falls occurred during daytime (85%), in a patient room (90%), and were unobserved (74%). About a half of all falls occurred during the first week of rehabilitation stay. Multivariate model revealed that diagnosis of stroke and amputation, age between 41 and 50 yrs, lower cognitive FIM scores, and a large number of medical comorbidities (> or =9) were associated with a high risk for fall. The respective prevalence ratios were 1.79, 3.80, 2.01, 0.98, and 1.50. CONCLUSIONS: The rate of falls varies considerably among different diagnostic groups admitted to inpatient rehabilitation. Mid-aged people with stroke and amputation, worse cognitive functions, and greater medical complexity are at a higher risk for falling. Admission FIM score may be of value for predicting falls in rehabilitation setting, which warrants further investigation. PMID- 18427219 TI - Functional and economic outcomes of cardiopulmonary patients: a preliminary comparison of the inpatient rehabilitation and skilled nursing facility environments. AB - OBJECTIVE: To examine the major clinical and economic outcomes of cardiopulmonary patients referred for inpatient rehabilitation or skilled nursing care after an acute care stay. DESIGN: Retrospective, exploratory study conducted at inpatient rehabilitation facilities (IRF) and a matched skilled nursing facility (SNF). Participants were cardiovascular and pulmonary patients (n = 495; 76.2 +/- 0.5 yrs). Measurements included changes in functionality (assessed by functional independence measure [FIM] and minimum data set [MDS]), length of stay (LOS), total and itemized facility charges, discharge disposition, and mortality. RESULTS: Participation in physical and occupational therapies occurred during 72 78% and 48-51% of total days in the IRF and SNF, respectively (P < 0.001). Changes in eating, grooming, bathing, dressing, toileting, bed-chair transfers, walking, verbal expression, problem solving, and auditory comprehension were greater in patients from the IRF than SNF (all P < 0.0001). LOS was longer in the SNF than IRF (34.7 +/- 3.4 vs. 14.9 +/- 0.5 days, P < 0.0001). In the IRF compared with the SNF, total charges ($22,162 vs. $10,873), pharmacy charges ($3104 vs. $1604), and combined physical and occupational therapy charges were higher ($5225 vs. $3582), all P < 0.0001. More IRF patients than SNF patients were discharged home (77.5% vs. 44.1%), and fewer IRF patients than SNF patients were discharged to acute care (15.8% vs. 23.2%) or expired (1.3% vs. 13.6%) during their stay (P < 0.05). CONCLUSIONS: More patients achieved functional independence, had shorter LOS, and had a higher rate of homebound discharge in the IRF than in the SNF, and this is associated with a higher cost of care. These SNF outcomes may be related to advancing age, type of illness, dementia, and inability to fully participate in therapies. PMID- 18427220 TI - Botulinum toxin type A injections into the calf muscles for treatment of spastic equinus in cerebral palsy: a randomized trial comparing single and multiple injection sites. AB - OBJECTIVE: To investigate the hypothesis that the multiple-site injection technique is associated with better outcomes than the single-point injection method in children with cerebral palsy and spastic equinus gait. DESIGN: A total of 17 children (nine boys, eight girls ages 1.8-9.4 yrs; eight hemiplegics, eight diplegics, one quadriplegic; levels I-IV with the Gross Motor Function Classification System) with 25 treated lower limbs were randomized into two groups: a single-point group receiving a standard dose of botulinum toxin A injection into one site and a multiple-points group into two sites on both heads of the gastrocnemius. Active and passive range of movement, selective dorsiflexion, dynamic muscle length (modified Tardieu scale), calf tone (modified Ashworth scale), attainment of anticipated gait pattern (Goal Attainment Scale), and video gait analysis (Observational Gait Scale [OGS]) were assessed before and 1, 2, and 4 mos after intervention. RESULTS: Both groups improved in dynamic muscle length, muscle tone, OGS-total scores and initial foot contact scores and a similar number of children attained their goals on the Goal Attainment Scale. The only significant difference between the groups was observed at 2 mos in passive dorsiflexion with the knee flexed, favoring the single-point group. Though not significantly, the incidence of adverse effects was higher in the multiple-points group. CONCLUSIONS: Using the methods described, no major changes in main outcome measures were associated with the number of injection sites. Issues other than efficacy guide the decision on whether to inject in single or multiple sites when treating spastic equinus with botulinum toxin. PMID- 18427222 TI - MRI artifact mimicking root compression by interbody cage displacement. PMID- 18427224 TI - RE: Effect of short-term hospitalization on functional capacity in patients not restricted to bed. PMID- 18427225 TI - Pain, mind, and movement: an expanded, updated, and integrated conceptualization. PMID- 18427226 TI - Toward a biopsychomotor conceptualization of pain: implications for research and intervention. AB - OBJECTIVES: Nearly 400 years ago, Rene Descartes proposed a model of pain perception that characterized pain as a purely physical phenomenon, devoid of psychologic influence. The characterization of pain as an exclusively sensory (or experiential) phenomenon continues to dominate current conceptualizations of pain. METHODS: This paper advances the view that the exclusive focus on pain sensation or experience as the essential feature of the pain system has given rise to conceptual frameworks that are incomplete and flawed. It is argued that individuals with pain differ from individuals without pain not only in how they "feel" but they differ in how they "behave." RESULTS: Arguments are put forward advocating for a biopsychomotor conceptualization of pain where pain behaviors are construed as integral components of the pain system. The biopsychomotor model proposes that at least 3 partially independent behavioral subsystems are integral components of pain. These include communicative pain behaviors, protective pain behaviors, and social response behaviors. Evidence is reviewed suggesting that different dimensions of pain behavior are functionally distinct, and questions are raised about the nature of motor programs responsible for the elicitation and maintenance of different forms of pain behavior. DISCUSSION: Clinical and theoretical implications of a biopsychomotor conceptualization of pain are discussed. PMID- 18427227 TI - Muscle pain: sensory implications and interaction with motor control. AB - Muscle hyperalgesia and referred pain plays an important role in chronic musculoskeletal pain. New knowledge on the involved basic mechanisms and better methods to assess muscle pain in the clinic are needed to revise and optimize the treatment regimes. Increased muscle sensitivity is manifested as (1) pain evoked by a normally non-nociceptive stimulus (allodynia), (2) increased pain intensity evoked by nociceptive stimuli (hyperalgesia), or (3) increased referred pain areas with associated somatosensory changes. Quantitative sensory testing provides the possibility to evaluate these manifestations in a standardized way in patients suffering from musculoskeletal pain or in healthy volunteers. Some manifestations of sensitisation, such as expanded referred muscle pain areas in chronic musculoskeletal pain patients, can be explained from animal experiments showing extrasegmental spread of sensitisation. An important part of the pain manifestations (eg, tenderness and referred pain) related to chronic musculoskeletal disorders may be due to peripheral and central sensitization, which play a role in the transition from acute to chronic pain. In recent years, it has become evident that muscle pain can interfere with motor control strategies and different patters of interaction are seen during rest, static contractions, and dynamic conditions. PMID- 18427228 TI - Virtual reality pain control during burn wound debridement in the hydrotank. AB - OBJECTIVE: Most burn-injured patients rate their pain during burn wound debridement as severe to excruciating. We explored the adjunctive use of water friendly, immersive virtual reality (VR) to distract patients from their pain during burn wound debridement in the hydrotherapy tank (hydrotank). SETTING: This study was conducted on inpatients at a major regional burn center. PATIENTS: Eleven hospitalized inpatients ages 9 to 40 years (mean age, 27 y) had their burn wounds debrided and dressed while partially submerged in the hydrotank. INTERVENTION: Although a nurse debrided the burn wound, each patient spent 3 minutes of wound care with no distraction and 3 minutes of wound care in VR during a single wound care session (within-subject condition order randomized). OUTCOME MEASURES: Three 0 to 10 graphic rating scale pain scores (worst pain, time spent thinking about pain, and pain unpleasantness) for each of the 2 treatment conditions served as the primary dependent variables. RESULTS: Patients reported significantly less pain when distracted with VR [eg, "worst pain" ratings during wound care dropped from "severe" (7.6) to "moderate" (5.1)]. The 6 patients who reported the strongest illusion of "going inside" the virtual world reported the greatest analgesic effect of VR on worst pain ratings, dropping from severe pain (7.2) in the no VR condition to mild pain (3.7) during VR. CONCLUSIONS: Results provide the first available evidence from a controlled study that immersive VR can be an effective nonpharmacologic pain reduction technique for burn patients experiencing severe to excruciating pain during wound care. The potential applications of VR analgesia to other painful procedures (eg, movement or exercise therapy) and other pain populations are discussed. PMID- 18427229 TI - Treatment expectancy and credibility are associated with the outcome of both physical and cognitive-behavioral treatment in chronic low back pain. AB - INTRODUCTION: Patients' initial beliefs about the success of a given pain treatment are shown to affect final treatment outcome. The Credibility/Expectancy Questionnaire (CEQ) has recently been developed as measure of treatment credibility and expectancy. OBJECTIVE: The objectives of this study were (1) to investigate the factor structure of the CEQ in a sample of chronic low back pain (CLBP) patients by means of a confirmatory factor analysis, (2) to examine the association between treatment credibility and expectancy and patient characteristics, and (3) to assess whether treatment expectancy and credibility are associated with the outcome of rehabilitation treatment. METHODS: CLBP patients (n=167) were randomized to either active physical therapy (n=51), cognitive-behavioral therapy (n=57), or a combination therapy (n=59), and completed the CEQ after a careful explanation of the treatment rationale. RESULTS: Confirmatory factor analysis supported the 2-factor structure (credibility/expectancy) of the CEQ. Lower credibility was associated with higher pain-related fear and lower internal control of pain, and lower expectancy with higher levels of pain-related fear and no radiating pain. Multiple linear regression analyses revealed that after controlling for age, sex, treatment center, pain-intensity at baseline, duration of disability, and irrespective of the treatment offered, expectancy was significantly associated with disability and satisfaction. Credibility was significantly associated with patient-specific symptoms and satisfaction. For global perceived effect, treatment expectancy was predictive in active physical therapy only, and treatment credibility was a significant predictor in combination therapy only. DISCUSSION: Although the associations found were low to modest, these results underscore the importance of expectancy and credibility for the outcome of different active interventions for CLBP and might contribute to the development of more effective treatments. PMID- 18427230 TI - Are we measuring what we need to measure? AB - Optimal treatment for patients with chronic pain remains elusive. A growing international consensus advocates evidence-based practice with assessment of clinical outcomes to improve the process and outcome of care. Clinical decision making about treatment options for an individual patient should include the patient's clinical presentation, available evidence, and patient preferences. Treatment should then be monitored and outcomes of treatment assessed. Although the placement of clinical decision making on a scientific, often quantitative basis as opposed to a subjective, impressionistic approach makes intuitive sense, the question is whether we have been measuring what we need to measure to practice evidence-based practice when we consider the current available evidence on pain management? The methods of synthesis of available evidence are still in development. Much of the evidence, although having internal validity, has limited external validity and is difficult to apply to the individual patient. Patients with chronic pain are a heterogeneous group, and different interventions may be indicated for different subgroups of patients. Various methods are being developed to better match patients with treatment. Little information exists on patient preferences, or how best to measure these. Information on how health care providers make clinical decisions is also scarce. Outcome measurement has come a long way and core domains to be measured have been established. Establishing normative data is a next main goal. Important methodologic and practical challenges remain to formulate evidence that can be applied to the individual patient with chronic pain. PMID- 18427231 TI - The restore rechargeable, implantable neurostimulator: handling and clinical results of a multicenter study. AB - BACKGROUND: Spinal cord stimulation is an effective therapy for chronic, neuropathic pain refractory to medication. Use of a rechargeable neurostimulation system (Restore, Medtronic Inc) could provide greater longevity in the treatment of complex pain. However, patients' ability to successfully recharge a neurostimulation system has not yet been demonstrated. PRIMARY OBJECTIVE: Ability of patients to recharge the neurostimulator. SECONDARY OBJECTIVES: Patient and physician satisfaction with the system, pain relief, quality of life, functional status, adverse events. METHODS: Prospective, open-label, multicenter, European study in patients with long-term refractory neuropathic pain. Recharging ability was assessed 1-month postimplant. Patient and physician satisfaction, pain relief, quality of life, and functional status were assessed at scheduled follow up visits through 12 months. Adverse events were monitored throughout. RESULTS: Primary end point: 100% of patients (n=41) successfully recharged the neurostimulator. Secondary end points at 1 month: 78.6% of patients found recharging easy. At 12 months: physicians were satisfied with the system for 92.7% of patients; pain intensity decreased significantly (P<0.001); mean self reported pain relief was 62%; 80.5% of patients had more than 50% pain relief; quality of life and functional status improved significantly (P<0.001); 98% of patients would recommend spinal cord stimulation to others. Overall, 41 device related complications (23 patients) were observed. CONCLUSIONS: Twelve-month experience indicates that the rechargeable neurostimulation system (Restore) was easy to use, with 100% of patients able to recharge successfully. Patient and physician satisfaction was high, with significant improvements in pain, quality of life, and functional status. Complications were comparable to prior experience with this therapy. PMID- 18427232 TI - Pain characteristics and their association with quality of life and self-concept in children with hemiplegic cerebral palsy identified from a population register. AB - OBJECTIVES: To describe and characterize the prevalence and quality of pain in a population-based community sample of children with hemiplegic cerebral palsy. METHODS: Outcomes were assessed from 2 domains of the World Health Organization International Classification of Functioning, Disability and Health: body structure/function (upper limb spans, modified Ashworth scale, Tardieu scale, sensory function), activities-participation (Assessment of Motor and Process Skills), Pediatric Quality of Life Inventory (PedsQL), and self-perception (Harter Self-Perception Profile). RESULTS: There were 107 participants of age (mean, 95% confidence interval) 8.94 years (8.23 to 9.65); 61 (57%) were boys and 58 (54%) had hemiplegia affecting the right side. Fifty-one (48%) reported pain. Pain mostly affected the involved side (n=28, 55%) and lower limb (n=19, 37%). Eighteen (35%) reported moderate to severe pain. Thirty-five (69%) said pain impacted on movement/activity and was of an aching quality (n=29, 57%). Mean (95% confidence interval) quality of life according to Pediatric Quality of Life Inventory was significantly lower for children experiencing pain compared with children experiencing no pain [parent 50.2 (45.9 to 54.5) vs. 60.1 (55.1 to 65.1), P<0.01; child 60.5 (55.4 to 65.6) vs. 75.8 (68.4 to 83.2), P<0.01]. The self-perception domains of Scholastic Competence [no pain, 3.02 (2.78 to 3.26); pain, 2.55 (2.31 to 2.79) P<0.01] and Behavioral Competence [no pain, 3.33 (3.07 to 3.60); pain, 2.88 (2.70 to 3.06) P<0.01] were significantly lower in children with pain aged > or =8 years. DISCUSSION: Pain is common in children with hemiplegic cerebral palsy with qualities suggesting a nociceptive origin. Pain is associated with lower quality of life and self-perception. Results suggest clinicians should assess and actively manage pain in this population. PMID- 18427233 TI - Experimental pain sensitivity in women with temporomandibular disorders and pain free controls: the relationship to orofacial muscular contraction and cardiovascular responses. AB - OBJECTIVE: Chronic pain may result both from a generalized hypersensitivity to acute pain, suggestive of central sensitization processes, and dysfunction of the endogenous pain regulatory system. One purpose of this study was to compare experimental pain sensitivity at several anatomic sites in temporomandibular disorder (TMD) patients and pain-free controls during baseline and after standardized mechanical load of the orofacial region. A second purpose was to compare the pain-modulating effects of cardiovascular responses in TMD patients and pain-free controls. METHODS: Experimental pain was induced by electrocutaneous stimulation of the dorsal left hand and pressure algometry at the right masseter muscle and the sternum. The pain sensitivity of the orofacial region was manipulated by isometric contraction of the masseter muscles. Elevations of mean arterial pressure and heart rate were induced by a simulated job interview. RESULTS: At baseline, the TMD patients exhibited a significantly higher electrocutaneous pain threshold. Relative to the healthy controls, the TMD patients reported increased electrocutaneous and pressure pain sensitivity after isometric contraction of the orofacial region. In addition, there were correlations between mean arterial pressure and pain sensitivity in the TMD group only. DISCUSSION: Significant increases in generalized pain sensitivity occurred in the TMD group, but not in the control group, after isometric contraction of the orofacial muscles, suggestive of a central sensitization process in TMD. Moreover, only in the TMD group there were significant associations between cardiovascular responsesand pain sensitivity, challenging previous assumptions of this relationship occurring mainly in pain-free individuals. PMID- 18427234 TI - Modulation of pain in osteoarthritis: the role of nitric oxide. AB - BACKGROUND: Patients with osteoarthritis (OA) may experience severe pain, progressive loss of movement function, and disability. Many pain-relieving medications are not effective, and are not able to improve the existing pathology. OBJECTIVES: This review summarizes (1) the pathology, mechanisms of pain production, and conservative management of OA with respect to pain; and (2) explains the role of nitric oxide (NO) in pain reduction and production, especially as related to OA. DISCUSSION: NO is produced in biologic cells by a family of enzymes referred to as the nitric oxide synthases (NOSs). The beneficial or harmful effects of different isoforms, constitutive NOS (cNOS) and inducible NOS (iNOS), respectively, suggest dual effects of NO in biologic structures. The harmful effects of NO are most often reported in the literature. We suggest that (1) NO via the beneficial cNOS pathway is decreased in joint structures exposed to chronic load-induced stresses and biochemical change induced stresses, (2) monochromatic infrared light energy at an 890 nm wavelength, applied at the skin surface, is absorbed into blood vessels and stimulates production of NO in joints by the beneficial cNOS pathway, (3) NO from the cNOS pathway may help decrease the detrimental effects of NO induced by iNOS and produced in OA pathology, and (4) NO-based intervention may produce substantial pain relief without undesirable side effects by increasing circulation, decreasing nerve irritation, and decreasing inflammation in joints. KEY MESSAGES: (1) The roles of NO in nociception are dual and complex. (2) NO via cNOS, produced transiently in small amounts, can bring dramatic relief to people with painful OA. PMID- 18427236 TI - Feasibility of a strength-training program to reduce pain in older women in assisted living. PMID- 18427235 TI - Post herpetic itching--a treatment dilemma. AB - OBJECTIVES: To present a case of severe disabling postherpetic itching (PHI) and discuss possible mechanisms and management. METHODS: We report on a 22-year-old male patient with a history of non-Hodgkin lymphoma, chronic renal failure peritoneal dialysis dependent, presented with a disabling pruritus around his left eye and forehead. Two months before, he was diagnosed with herpes zoster ophtalmicus. His itching intensity was 10/10 on a visual analog scale and he reported no pain. The neurologic examination showed a hyposensitivity to touch around his left eye. RESULTS: Our patient suffered of PHI who responded successfully to a combination of antihistamine and an antiepileptic agent. DISCUSSION: The mechanism of postherpetic neuralgia and PHI are not well understood and no single best treatment for postherpetic neuralgia and PHI is known. Clinical experience suggested that neuropathic itch may be more resistant to treatment than neuropathic pain. This immunocompromized patient with a severe disabling PHI responded to antihistaminic and anticonvulsant treatment. PMID- 18427238 TI - Current opinions on selected respiratory infections causing global morbidity and mortality. PMID- 18427239 TI - Probiotics for the prevention of nosocomial pneumonia: current evidence and opinions. AB - PURPOSE OF REVIEW: Nosocomial infections are common and are associated with considerable morbidity and mortality. The continuing evolution of multidrug resistant pathogens and ineffective therapy for the infections they cause has stimulated interest in the potential of probiotic products to prevent nosocomial infections. Probiotics are viable microorganisms that colonize the host and exert antibacterial and immunomodulatory effects. This article will review the current evidence for probiotics in preventing nosocomial infections, particularly pneumonia in a diverse population of critically ill patients. RECENT FINDINGS: Currently, there are insufficient data to conclusively determine whether probiotics are beneficial in the prevention of nosocomial infections, particularly nosocomial pneumonia. Most of the current literature is limited by poor trial design, inadequate blinding, small study samples, and poorly defined endpoints. SUMMARY: Probiotic products reduce pathogenic colonization of the host. Despite the theoretical plausibility, there is currently insufficient evidence that probiotic products reduce the incidence of nosocomial pneumonia. Large, multicenter, randomized clinical trials utilizing a rigorous, invasive diagnostic approach to nosocomial pneumonia need to be performed to prospectively evaluate the utility of probiotic products. In addition, bench research needs to be performed to select the most appropriate probiotic formulation for different clinical applications. PMID- 18427240 TI - Early diagnosis of lower respiratory tract infections (point-of-care tests). AB - PURPOSE OF REVIEW: Respiratory tract infections are a common reason for prescribing antibiotics, although not all of these infections require such therapy. Rapid diagnosis of etiology using point-of-care tests is a potentially useful way of reducing prescriptions of both unnecessary and unnecessarily broad spectrum antibiotics. This can also lead to the facilitation of appropriate infection control measures to prevent spread of respiratory viruses within institutions. RECENT FINDINGS: Point-of-care tests are available for diagnosing influenza, respiratory syncytial virus, Streptococcus pneumoniae, and Legionella infections using easily obtainable specimens. Their main benefit is that results can be obtained in about 15 min with reasonable accuracy. In many situations, however, it is still important to confirm diagnosis with more accurate but slower tests such as bacterial cultures with antibacterial susceptibility testing or viral polymerase chain reaction testing. SUMMARY: Although the sensitivities of many of the rapid diagnostic tests are moderate, when used at the time of initial consultation, they have the potential to reduce costs, length of stay, secondary spread of respiratory viruses, and inappropriate antibiotic prescribing. PMID- 18427241 TI - Atypical pneumonias: current clinical concepts focusing on Legionnaires' disease. AB - PURPOSE OF REVIEW: This review provides clinicians with an overview of the clinical features of the atypical pneumonias. Atypical community-acquired pneumonia pathogens cause systemic infections with pneumonia. The key to the clinical diagnosis of atypical pneumonias depends on recognizing the characteristic pattern of extrapulmonary organ involvement different for each pathogen. As Legionella is likely to present as severe pneumonia and does not respond to beta-lactams, it is important to presumptively diagnose Legionnaires' disease clinically so that Legionella coverage is included in empiric therapy. This study reviews the clinical features and nonspecific laboratory markers of atypical pathogens, focusing on Legionnaires' disease. RECENT FINDINGS: Case reports/outbreaks increase our understanding of Legionnaires' disease transmission. Both Mycoplasma pneumoniae and Chlamydophilia pneumoniae may cause asthma. Antimicrobial therapy of Chlamydophilia pneumoniae/Mycoplasma pneumoniae is important to decrease person-to-person spread and to decrease potential long term sequelae. SUMMARY: Atypical pulmonary pathogens cause systemic infections accompanied by a variety of characteristic extrapulmonary features. Clinically, it is possible to differentiate Legionnaires' disease from the other typical/atypical pneumonias. Rapid clinical diagnosis of atypical pathogens, particularly Legionnaires' disease, is important in selecting effective empiric therapy and prompting definitive laboratory testing. PMID- 18427242 TI - Aetiology, diagnosis and management of infective causes of severe haemoptysis in intensive care units. AB - PURPOSE OF THE REVIEW: Infective causes of severe haemoptysis have progressively shifted to causes related to chronic inflammatory lung diseases. Physicians should, however, recognize the most common of them, for example necrotizing parenchymal infections, tuberculosis and mycetoma. RECENT FINDINGS: The recent increase in the incidence of a devastating Panton-Valentine leukocidin-associated staphylococcal pneumonia has reminded us of the crucial role of prompt diagnosis and management. General supportive care should be administered to prevent asphyxiation in addition to starting appropriate antibiotics as soon as possible. Once the bleeding has been controlled, the diagnostic strategy should integrate a detailed medical history, physical examination, Gram stain of the respiratory specimens and chest radiograph. Computed tomography scan has dramatically improved the diagnosis and the treatment of infective causes of severe haemoptysis by assessing the cause and mechanism(s) of haemoptysis. Although bronchial arteries are the major source of bleeding, nonbronchial systemic and pulmonary arteries' involvement should be feared, especially in haemoptysis related to tuberculosis and mycetoma. SUMMARY: Endovascular therapy should be first attempted to control the bleeding and then elective surgery performed in case of localized lesion and adequate pulmonary function. Fibreoptic bronchoscopy with broncho-alveolar lavage remains the cornerstone of diagnosis in immunocompromised hosts with haemoptysis and in the rare cases of alveolar haemorrhage related to infectious diseases. PMID- 18427243 TI - Nontuberculous mycobacterial pulmonary disease. AB - PURPOSE OF REVIEW: Pulmonary disease caused by nontuberculous mycobacteria is occurring with greater frequency, and previously unrecognized manifestations of nontuberculous mycobacteria are being identified. Paralleling this increase, improvements in laboratory techniques now allow for more precise identification of nontuberculous mycobacteria and recognition of new species. Consequently, clinicians are more often confronted with diagnostic and therapeutic challenges relevant to the care of patients with nontuberculous mycobacterial lung disease. RECENT FINDINGS: In response to this burgeoning clinical need, the American Thoracic Society and Infectious Disease Society of America jointly published an updated consensus statement on nontuberculous mycobacterial pulmonary disease in 2007. This document, in conjunction with original investigations in the field, has advanced our understanding of the pathogenesis of nontuberculous mycobacterial lung disease, its clinical manifestations, and the efficacy of medical and surgical therapy. SUMMARY: The present article will review our current understanding of nontuberculous mycobacterial pulmonary disease with particular emphasis on pathogenesis, diagnosis, and therapeutic decision making. Areas of clinical controversy in which current data are inadequate to guide our decision making will be highlighted. PMID- 18427244 TI - Fungal infections in lung transplant recipients. AB - PURPOSE OF REVIEW: Despite numerous advances in lung transplantation, survival is still significantly compromised by a higher rate of infections. This review intends to provide an overview of the most common fungal infections afflicting lung transplant recipients, focusing on the most recent developments in diagnosis, therapy and prophylaxis. RECENT FINDINGS: Although detection of galactomannan in serum has poor sensitivity for the diagnosis of invasive aspergillosis in lung transplant recipients, detection of galactomannan in the bronchoalveolar lavage of a lung transplant recipient, with a compatible clinical illness, is highly suggestive of invasive disease. New antifungal agents, with a broader spectrum of activity and a more tolerable side effect profile, have become available for the treatment and prophylaxis of fungal infections. Evidence suggests that, at least for voriconazole, monitoring of drug concentrations may be advisable to prevent clinical failure due to underdosing and toxicity due to excessive dosing. SUMMARY: Fungal infections remain a significant cause of morbidity and mortality in lung transplant recipients; however, advances have been made in the recent years, which will allow earlier diagnosis and more effective and tolerated treatment. PMID- 18427245 TI - Pulmonary nocardiosis: risk factors, clinical features, diagnosis and prognosis. AB - PURPOSE OF REVIEW: Pulmonary infection by Nocardia spp. has been recognized for the past 100 years. The number of cases of pulmonary nocardiosis reported in the literature is increasing, and in some cases, a diagnosis is reached even postmortem. This increase is partly due to the growing number of patients with depressed cellular immunity. The diagnosis of this infection, which has a high rate of mortality, is usually delayed, due to a nonspecific clinical-radiological presentation and the difficulties in cultivating the bacteria. This review analyzes the current situation, in order to better understand this infection and enhance awareness and clinical suspicion that would lead to further specific microbiological studies and treatment. RECENT FINDINGS: The number of case series in the literature is increasing. This development may be due to an absolute increase in the number of immunocompromised patients, but also to improvements in laboratory techniques and molecular methods to detect nocardiosis. SUMMARY: It is important to keep in mind, and suspect the presence of pulmonary nocardiosis in the immunodepressed patients with clinical pulmonary infection. Furthermore, the laboratory should be notified when Nocardia is suspected in a clinical specimen so that measures can be taken to optimize recognition and recovery of the organism. PMID- 18427246 TI - Advances in the biology, pathogenesis and identification of Pneumocystis pneumonia. AB - PURPOSE OF REVIEW: Pneumocystis pneumonia remains the most prevalent opportunistic infection in patients with AIDS. It is also a common devastating infection in patients with other causes of altered immunity. Though scientific study of this fungal pathogen is challenging given the inability to propagate the organism outside of the host lung, studies utilizing advanced molecular techniques and genomic analysis have broadened our understanding of the epidemiology and pathogenesis of Pneumocystis and will be described herein. RECENT FINDINGS: Results from advanced molecular techniques suggest that Pneumocystis organisms not only cause infection in patients with impaired immunity but also colonize mammals with normal immune systems. Advanced technology has also identified acquired Pneumocystis genetic mutations that confer resistance to currently utilized therapeutics. Though not yet widely utilized in clinical medicine, advanced polymerase chain reaction techniques improve the diagnostic yield of respiratory specimen analysis. Preliminary results from serum beta-glucan testing suggest that a noninvasive marker of Pneumocystis pneumonia infection and response to therapy may be on the horizon. SUMMARY: Recent scientific advances suggest opportunities for improving the diagnosis and treatment surveillance of Pneumocystis pneumonia. Further investigations are necessary to define the optimal characteristics of these laboratory tests and to develop therapeutics directed at novel Pneumocystis genomic targets. PMID- 18427247 TI - Respiratory syncytial virus infection in adults. AB - PURPOSE OF REVIEW: Respiratory syncytial virus has increasingly been recognized as a clinically significant cause of respiratory tract infections in adults, especially among the elderly and the immunocompromised. RECENT FINDINGS: Advances in molecular diagnostic methods have enabled rapid diagnosis of respiratory syncytial virus for clinical and epidemiological studies. Recent reports focus on clinical, immunological, and/or radiographic characterizations of respiratory syncytial virus infection in adults, particularly in hospitalized patients and those with underlying chronic obstructive pulmonary disease, and therapeutic and prophylactic use of antiviral agents in immunocompromised adults. Respiratory syncytial virus vaccine development remains a high priority, with the testing of genetically engineered live attenuated vaccines leading to further insights into the pathogenesis of respiratory syncytial virus in adults. SUMMARY: Further studies are necessary to elucidate the pathogenesis and immune response against respiratory syncytial virus in adults. The significant burden of respiratory syncytial virus-induced disease in adults and the limited number of approved antiviral agents reinforce the need to develop a respiratory syncytial virus vaccine. PMID- 18427248 TI - Clinical features, pathogenesis and immunobiology of severe acute respiratory syndrome. AB - PURPOSE OF REVIEW: Severe acute respiratory syndrome coronavirus is a novel virus responsible for the major pandemic in 2003, and it re-emerged in China in late 2003 and 2004 following resumption of wild animal trading activities. Over the past few years, research work has looked into factors that may lead to super spreading events, clinical/laboratory parameters that may differentiate severe acute respiratory syndrome from other causes of community-acquired pneumonia, the origin of severe acute respiratory syndrome, reservoir host distribution and transmission routes. A better understanding of these issues may help prevent and control future outbreaks of severe acute respiratory syndrome. RECENT FINDINGS: Bats are natural reservoirs of severe acute respiratory syndrome like coronaviruses. The human and civet isolates of severe acute respiratory syndrome coronavirus nestle phylogenetically within the spectrum of severe acute respiratory syndrome like coronaviruses. Severe acute respiratory syndrome has the potential of being converted from droplet to airborne transmission. When evaluating epidemiologically high-risk patients with community-acquired pneumonia and no immediate alternative diagnosis, a low absolute neutrophil count on presentation, along with poor responses after 72 h of antibiotic treatment, may raise the index of suspicion for severe acute respiratory syndrome. SUMMARY: The presence of severe acute respiratory syndrome like coronaviruses in horseshoe bats raises the possible role of bats in previous and potentially future severe acute respiratory syndrome outbreaks in human. Healthcare workers should take adequate respiratory protection in addition to strict contact and droplet precautions when managing patients with severe acute respiratory syndrome. PMID- 18427249 TI - Recent antiviral strategies against human coronavirus-related respiratory illnesses. AB - PURPOSE OF REVIEW: The main purpose of this review is to summarize the current research (2006-2007) concerning the development of novel anticoronaviral strategies and compounds. RECENT FINDINGS: Recent research led to the identification of several novel agents inhibiting coronaviral replication. The most promising compounds include carbohydrate-binding agents, neutralizing antibodies and drugs targeting a coronaviral envelope protein. SUMMARY: Although initial outbreaks of coronavirus that causes severe acute respiratory syndrome (SARS-CoV) were controlled by public health measures, the development of vaccines and antiviral agents for SARS-CoV is essential for improving control and treatment of future outbreaks. Four years after the SARS-CoV epidemic, several compounds with an anticoronaviral activity have been identified. PMID- 18427250 TI - Surgery for pulmonary tuberculosis. AB - PURPOSE OF REVIEW: The role of thoracic surgery in pulmonary tuberculosis, in either a diagnostic or a therapeutic role, is well established. This review aims to underline the current indications for lung resection, surgical strategy and outcomes associated with surgery. RECENT FINDINGS: There has been a renewed interest in surgery, mainly in the multidrug-resistant tuberculosis (MDRTB) group as an adjunct to medical therapy. Lung resection in this group of patients has been undertaken with acceptable morbidity and mortality, and with cure rates of over 90%. The current strategy favours early surgical intervention for patients with multidrug-resistant tuberculosis to prevent the development of bilateral disease. Early surgery for haemoptysis is advocated, for sometimes it is necessary to operate on these patients without a full course of antituberculous therapy. Medical stabilization and early inpatient surgery is favoured in view of the excessive mortality associated with emergency surgery. There is still no consensus on the management of the bronchial stump at the time of lung resection, although the current trend favours closure, either sutured or stapled, without muscle reinforcement. SUMMARY: Lung resection for pulmonary tuberculosis, in carefully selected and prepared patients, is safe with acceptable morbidity and mortality. PMID- 18427251 TI - Pulmonary defense mechanisms against pneumonia and sepsis. AB - PURPOSE OF REVIEW: Pneumonia and sepsis are major causes of morbidity and mortality. Lung defense against pathogens involves innate and acquired immune responses. In this review, we focus on lung pathogens associated with sepsis and the innate immune response to them. In addition to discussing typical lung pathogens, the structural defenses, antimicrobial particles, complement, and cellular components of the immune response against these pathogens are also explored. RECENT FINDINGS: The rising importance of pathogen-associated molecular pattern recognition molecules (such as Toll-like receptors) is discussed, as is the pivotal role of the dendritic cell in linking the innate and adaptive immune response. Although the adaptive response is delayed, it is more specific than the innate response and confers long-lasting memory allowing rapid and efficient clearance of pathogens on subsequent re-exposure. Sepsis interferes with both the innate and adaptive immune responses by inducing marked apoptosis of dendritic cells and lymphocytes, suppressing both arms of the immune response. This immunosuppression is a major hallmark of this disorder. SUMMARY: Clearly, understanding the immune response in the lung is critical to the development of future therapeutics for pneumonia and sepsis. This review concludes with novel discoveries in the lung immune response, which may lead to future avenues of treatment. PMID- 18427252 TI - Bibliography. Current world literature. Infectious diseases. PMID- 18427254 TI - Pediatric ophthalmology. Preface. PMID- 18427255 TI - Cataract surgery in children with juvenile idiopathic arthritis associated uveitis. PMID- 18427256 TI - Tubulointerstitial nephritis and uveitis syndrome. PMID- 18427257 TI - Blepharophimosis-ptosis-epicanthus inversus syndrome (BPES): clinical manifestation and treatment. PMID- 18427258 TI - Update on pediatric keratoplasty. PMID- 18427259 TI - Anterior segment dysgenesis: Peters anomaly and sclerocornea. PMID- 18427260 TI - The role of soft contact lenses as an adjunct to the Boston keratoprosthesis. PMID- 18427261 TI - The vitreo-retinal manifestations of persistent hyperplasic primary vitreous (PHPV) and their management. PMID- 18427262 TI - Surgical management of glaucoma with the sturge weber syndrome. PMID- 18427263 TI - Aniridia. PMID- 18427264 TI - Aphakic glaucoma after congenital cataract surgery. PMID- 18427265 TI - Abnormal eye movements in children. PMID- 18427266 TI - Diagnosis, classification, and treatment of retinoblastoma. PMID- 18427267 TI - Coats' disease. PMID- 18427268 TI - Inherited proliferative vitreoretinopathies of childhood. PMID- 18427269 TI - Infantile cataracts. PMID- 18427270 TI - Safety and efficacy of PRK and LASIK in pediatric patients. PMID- 18427271 TI - Herpetic simplex eye disease in the pediatric population. PMID- 18427272 TI - The etiology of V pattern strabismus in patients with craniosynostosis. PMID- 18427273 TI - Retinopathy of prematurity update. PMID- 18427274 TI - Shaken baby syndrome: a review and update on ophthalmologic manifestations. PMID- 18427276 TI - Effects of modifying triglycerides and triglyceride-rich lipoproteins on cardiovascular outcomes. AB - Elevated levels of triglycerides (and triglyceride-rich lipoproteins) are increasingly being recognized as treatment targets to lower cardiovascular risk in certain patient subgroups, including individuals receiving HMG-CoA reductase inhibitors (statins). Evidence suggests that these agents reduce the risk of coronary events more markedly in patients with elevated triglycerides and low levels of high-density lipoprotein cholesterol (HDL-C). Further, intensive long term statin therapy that reduces both low-density lipoprotein cholesterol (LDL-C) to <70 mg/dL and triglycerides to <150 mg/dL results in a decreased risk of cardiovascular events compared with more moderate statin treatment. Long-term therapy with fibric-acid derivatives, which lower triglycerides and raise HDL-C, appears to reduce mortality in patients with elevated triglycerides and/or those experiencing the most marked reductions in triglycerides on therapy. However, randomized clinical trials involving fibrates have not shown consistent benefit. Niacin (nicotinic acid), which is the most effective available medication for raising HDL-C and also lowers triglycerides, has not been as extensively studied as fibrates in long-term randomized controlled trials. Initial reports (eg, Coronary Drug Project) demonstrated a reduction in coronary disease but not total mortality in patients randomized to niacin. However, a 15-year follow-up demonstrated that all-cause mortality was significantly reduced in those initially randomized to niacin. At the pathophysiologic level, elevated triglycerides and triglyceride-rich lipoproteins are recognized as potential factors in driving atherosclerotic progression, particularly in mild-to-moderate lesions. Elevated triglycerides also constitute a plausible therapeutic target in certain patients with coronary heart disease (and/or insulin resistance) but without profound LDL-C elevations. The foregoing and other evidence has led consensus panels to lower the upper limit for "normal" triglycerides to 150 mg/dL. Adequately powered randomized controlled trials that specifically assess the effects of lowering triglycerides and raising HDL-C, and trials that target individuals with high triglycerides and low HDL-C, may provide data for recommending specific treatment targets for triglycerides and HDL-C, as well as effective and well-tolerated therapies to achieve these goals. PMID- 18427277 TI - Novel transient outward and ultra-rapid delayed rectifier current antagonist, AVE0118, protects against ventricular fibrillation induced by myocardial ischemia. AB - AVE0118 is a novel drug that blocks the transient outward current (Ito), the ultra rapid component of the delayed rectifier current (IKur), and the acetylcholine dependent potassium channel (IKach). The latter 2 channels are more abundant in atrial tissue. It is possible that AVE0118 could reduce regional differences in repolarization and thereby prevent malignant arrhythmias provoked by ischemia. To test this hypothesis, ventricular fibrillation was induced by a 2 minute occlusion of the left circumflex coronary artery during the last min of exercise in dogs with healed myocardial infarctions (n = 9). On a subsequent day, this exercise plus ischemia test was repeated after pretreatment with AVE0118 (1.0 mg/kg, IV). AVE0118 did not change QTc (Van de Water's correction) interval [245 +/- 6.0 ms (control) versus 242 +/- 2.3 ms (AVE)] and attenuated the dispersion of repolarization as measured by the duration of the descending portion of the T wave (Tpeak - Tend) induced by ischemia [ischemic changes: +11.1 +/- 2.4 ms (no drug) versus +2.2 +/- 3.7 ms (AVE)]. AVE0118 also significantly reduced the incidence of ventricular fibrillation, protecting 7 of 9 animals. Thus, AVE0118 abolished ischemically induced repolarization abnormalities and prevented malignant arrhythmias induced by ischemia without altering QTc interval. PMID- 18427278 TI - Tissue 65Zinc translocation in a rat model of chronic aldosteronism. AB - Zinc, an essential micronutrient, is involved in wound healing. The hypozincemia seen with chronic aldosteronism is associated with enhanced fecal and urinary excretory Zn losses, and its tissue distribution is less certain. This study monitored tissue 65Zn distribution in uninephrectomized rats at weeks 1 and 4 of aldosterone/salt treatment (ALDOST). Plasma and tissue total radionucleotide uptake was determined by calculating its mean radioactivity at 1, 4, 8, 24, and 48 hours after intravenous 65Zn administration and where respective area under the concentration-time curves (AUC) were determined by the linear trapezoidal rule and expressed as a tissue:plasma AUC ratio. Examined tissues included: (1) injured heart and kidney in response to ALDOST and incised skin; (2) noninjured liver, skeletal muscle, and spleen sites of stress-linked Zn uptake; and (3) bone, a major storage and release site when Zn homeostasis is threatened. In comparison with age-matched and gender-matched controls, the following were found with week 1 and 4 ALDOST: (1) reduced plasma 65Zn; (2) an accumulation of 65Zn in heart and kidneys, where a well-known vasculopathy involves intramural vessels, and in incised skin at week 1; (3) an organ-specific increase in tissue 65Zn in liver, in keeping with upregulated metallothionein expression, skeletal muscle, and spleen; and (4) a fall in bone and healed skin Zn at week 4. Thus a wide ranging disturbance in Zn homeostasis appears during ALDOST to include its translocation from plasma to injured heart, kidneys, and skin and noninjured liver, skeletal muscle, and spleen together with a resorption of stored Zn in bone at week 4. Zinc dyshomeostasis is an integral feature of chronic aldosteronism. PMID- 18427279 TI - Rapid stabilization of vulnerable carotid plaque within 1 month of pitavastatin treatment in patients with acute coronary syndrome. AB - We determined time course of stabilization of echolucent carotid plaques by statin therapy in patients with acute coronary syndrome (ACS). Treatment with 4 mg/d pitavastatin (n = 33) or placebo (n = 32) was initiated within 3 days after onset of ACS in 65 patients with echolucent carotid plaque. Vulnerable carotid plaques were assessed by measuring plaque echolucency using carotid ultrasound with integrated backscatter (IBS) analysis before and 1 month after treatment in all patients. The calibrated IBS value (intima-media IBS value minus adventia IBS) of vulnerable carotid plaques favorably changed at 1 month after treatment in both groups, but the echolucency at 1 month improved more in the pitavastatin than in the placebo group (pitavastatin group: -18.7 +/- 3.3 dB at pretreatment versus -12.7 +/- 2.3 dB at 1 month *P < 0.001; placebo: -19.0 +/- 3.5 dB versus 16.9 +/- 3.2 dB, P < 0.05, *P < 0.01 versus the value at 1 month in placebo group). Levels of CRP, VEGF, and TNFalpha at 1 month were significantly lower in pitavastatin than placebo group. In conclusion, pitavastatin improved carotid plaque echolucency within 1 month of therapy in patients with ACS, in association with decrease in the inflammatory biomarkers related to vulnerable plaques. PMID- 18427280 TI - A novel partial fatty acid oxidation inhibitor decreases myocardial oxygen consumption and improves cardiac efficiency in demand-induced ischemic heart. AB - The benefits of inhibition of fatty acid oxidation (FOX) and stimulation of glucose oxidation (GOX) in ischemia are controversial. The objective of this study was to evaluate the effect of the FOX inhibitor CVT-4325 on the rates of FOX, GOX, myocardial oxygen consumption (MVO2), and cardiac efficiency in the absence and presence of palmitate during demand-induced ischemia of the rodent isolated hearts. Palmitate concentration-dependently increased FOX, decreased GOX, and increased MVO2. CVT-4325 inhibited FOX and increased GOX in the presence (but not the absence) of 1.2 mM palmitate, with EC50 values of 0.9 and 5.8 microM, respectively. The potency for CVT-4325 to inhibit FOX was 10-fold greater (0.9 versus 9.7 microM) in the presence of 1.2 mM compared with 0.4 mM palmitate. The increase in MVO2 caused by 1.2 mM palmitate was significantly reduced by 3 to 10 microM CVT-4325 in guinea pig hearts. In the presence of 1.2 mM palmitate, an increase in pacing rate of the guinea pig heart from 3.5 to 6.5 Hz caused a significant 50% increase in MVO2, a decrease in cardiac efficiency, and an increase in lactate concentration in the cardiac effluent from 0.04 +/- 0.01 to 0.10 +/- 0.02 mM (P < 0.01). CVT-4325 (3 microM) attenuated the increase (P < 0.05) in MVO2 while maintaining cardiac contractility, and decreased the lactate production to 0.05 +/- 0.01 mM (P < 0.01). Thus, the FOX inhibitor CVT-4325 decreased MVO2 and increased myocardial efficiency during demand-(pacing)-induced ischemia in the presence of palmitate in the rodent isolated hearts. PMID- 18427281 TI - Characterization of human cardiac Kv1.5 inhibition by the novel atrial-selective antiarrhythmic compound AVE1231. AB - OBJECTIVE: Atrial-selective drug therapy represents a novel therapeutic approach for atrial fibrillation management. The aim of the present study was to investigate the mechanism of hKv1.5 channel inhibition by the atrial-selective compound AVE1231. METHODS: Ionic currents were recorded from CHO cells transfected with KCNA5 cDNA with whole-cell patch-clamp technique. The effect of AVE1231 on human atrial cell action potentials was explored with a computer model. RESULTS: KCNA5 expression resulted in typical K currents that activated and inactivated voltage dependently. Ascending concentrations of AVE1231 (0.1-100 microM) led to concentration- and voltage-dependent current inhibition (IC50 at +40 mV: 2.0 +/- 0.5 microM, Hill coefficient 0.69 +/- 0.12). Acceleration of hKv1.5 current inactivation occurred with increasing AVE1231 concentrations, indicating channel inhibition in the open state (eg, taufast at +40 mV: 318 +/- 92 milliseconds under control; 14 +/- 1 milliseconds with 3 microM, P < 0.05). Using 1/taufast as an approximation of the time course of drug-channel interaction, association rate (K+1) and dissociation rate (K-1) constants were 8.18 x 10 M/s and 45.95 seconds, respectively (KD = 5.62 microM). The onset of current inhibition occurred more rapidly with higher concentrations along with a prominent tail current crossover phenomenon after AVE1231 application. Drug inhibition remained effective through a range of stimulation frequencies. Computer modeling suggested more pronounced prolongation of action potential duration under conditions of atrial remodeling. CONCLUSION: AVE1231 is an inhibitor of hKv1.5 currents with predominant action on channels in their open state; thus, it may be suitable for the treatment of AF. PMID- 18427282 TI - Atorvastatin inhibits ABCA1 expression and cholesterol efflux in THP-1 macrophages by an LXR-dependent pathway. AB - The effect of atorvastatin on adenosine triphosphate (ATP)-binding cassette transporter A1 (ABCA1) expression and cholesterol efflux remains controversial. In an effort to clarify this issue, ABCA1 expression and apolipoprotein AI (apoAI)-mediated cholesterol efflux after atorvastatin treatment were investigated in THP-1 macrophages. Atorvastatin from 2 microM to 40 microM dose dependently inhibited ABCA1 expression in human monocyte-derived macrophages and phorbol 12-myristate 13-acetate (PMA)-stimulated THP-1 monocytes. ApoAI-mediated cholesterol efflux was reduced in PMA-stimulated THP-1 cells treated with atorvastatin, this effect was abolished with acetylated low-density lipoprotein (LDL) pretreatment. Atorvastatin treatment also dose-dependently reduced liver X receptor alpha (LXRalpha) expression and Rho activation. Rho activation by farnysylpyophosphate (FPP) and lysophosphatidic acid (LPA) did not salvage, but further depressed, the cholesterol efflux and ABCA1 expression in the presence of atorvastatin. Without atorvastatin, Rho activation by mevalonate, FPP, and LPA diminished apoAI-mediated cholesterol efflux, and Rho activation by GTPgammaS also decreased ABCA1 messenger ribonucleic acid (mRNA) by 16%. Furthermore, Rho inhibition by C3 exoenzyme increased ABCA1 mRNA by 48% despite a 17% decrease in apoAI-mediated cholesterol efflux. LXRalpha agonists (T01901317 and 22(R) hydroxycholesterol) prevented any reductions in cholesterol efflux or ABCA1 expression associated with atorvastatin treatment. Furthermore, Western blot analysis demonstrated the reciprocal inhibition of Rho and LXRalpha. In conclusion, atorvastatin decreases ABCA1 expression in noncholesterol-loaded macrophages in an LXRalpha- but not Rho-dependent pathway; this effect can be compromised after acetylated LDL cholesterol loading. PMID- 18427283 TI - Sodium tanshinone IIA sulfonate protects cardiomyocytes against oxidative stress mediated apoptosis through inhibiting JNK activation. AB - Sodium tanshinone IIA sulfonate (STS) is a water-soluble derivative of tanshinone IIA, a well-known Chinese medicine for treating cardiovascular disorders. Cardiomyocyte apoptosis plays a major role in the development of cardiovascular diseases. The present study was designed to investigate the effects of STS on cardiomyocyte apoptosis induced by in vivo acute myocardial infarction (MI) in adult rats and by in vitro H2O2-treated neonatal rat ventricular myocytes. In MI rats, STS significantly reduced the infarct sizes, the blood lactate dehydrogenase (LDH) level, and the number of apoptotic cardiomyocytes in the infarcted hearts. In the in vitro study, STS reversed the decreased effect of cell viability induced by H2O2. In addition, STS also markedly inhibited H2O2 induced cardiomyocyte apoptosis. C-Jun N-terminal kinases/stress-activated protein kinases (JNKs/SAPKs) and p38 MAPK are classic oxidative stress-activated protein kinases. Our further mechanistic study revealed that increased JNK phosphorylation stimulated by H2O2 was abolished by STS treatment. In conclusion, inhibition of JNK activation plays a significant role in cardioprotective effects of STS. PMID- 18427284 TI - Effects of antihypertensive drugs on capillary rarefaction in spontaneously hypertensive rats: intravital microscopy and histologic analysis. AB - We investigated the effects of chronic oral antihypertensive treatment on functional and structural capillary rarefaction in spontaneously hypertensive rats (SHR). Wistar Kyoto rats (WKY) were used as a normotensive control group. In untreated rats, intravital videomicroscopy showed that functional capillary density was lower in SHR skeletal muscle (WKY 395 +/- 17 and SHR 258 +/- 13 capillaries/mm, P < 0.01) and ear skin (WKY 391 +/- 18 and SHR 210 +/- 15 capillaries/mm, P < 0.01). A linear relationship was seen between skeletal muscle and skin capillary densities (r = 0.654, P < 0.0001). Histologic analysis showed that SHR had a lower capillary-to-fiber ratio in the skeletal muscle (WKY 1.74 +/ 0.08 and SHR 1.40 +/- 0.06, P < 0.01). Capillary volume density-to-fiber volume density ratio in the left ventricle of SHR was also reduced (WKY 0.55 +/- 0.09 and SHR 0.42 +/- 0.09, P < 0.01). The animals were treated with the angiotensin converting enzyme (ACE) inhibitor enalapril, the angiotensin II type I receptor (AT1) receptor antagonist losartan, the beta-blocker atenolol, or the calcium channel blocker nifedipine, resulting in similar reductions in systolic blood pressure (19.8%, 19.1%, 17.4%, and 18.2%, respectively, P > 0.05). Atenolol did not induce any change in functional capillary density of SHR. Losartan and nifedipine completely reversed functional capillary rarefaction in both muscle and cutaneous tissues, whereas enalapril significantly increased functional capillary density only in the skin. The skeletal muscle capillary-to-fiber ratio was normalized by enalapril, losartan, and nifedipine. Treatments with enalapril or losartan normalized the cardiac structural capillary rarefaction of SHRs, whereas atenolol and nifedipine had no effect. Our results suggest that different pharmacologic classes of antihypertensive drugs with similar effect on blood pressure differ in terms of their effect on the microcirculation. PMID- 18427285 TI - Pharmacology and safety of tetradecylthioacetic acid (TTA): phase-1 study. AB - This study describes the clinical, hematological, and biochemical safety of tetradecylthioacetic acid (TTA). A total of 18 healthy volunteers were included. Subjects were randomly assigned into 3 groups according to the daily given dose of TTA: group 1 (200 mg), group 2 (600 mg), and group 3 (1000 mg). TTA was given as a single oral dose for 7 consecutive days. Safety was evaluated by following the adverse events, vital signs, and hematological and biochemical parameters in blood and urine samples. Efficacy was estimated through its effects on plasma lipids profile. Few adverse events of mild severity were reported. No clinically significant changes were observed in the hematological or clinical chemical parameters in blood/urine. TTA did not induce significant changes in the blood lipids or free fatty acids, but it did result in an increase in plasma concentration of Delta9 desaturated TTA (TTA: 1n-8). Serum concentration pattern of TTA at day 1 showed a 1.5-hour lag time followed by a rapid absorption and a slower elimination phase. The median peak values were 2.9 mg/L (range, 1.1 to 5.4 mg/L), 11.5 mg/L (range, 4 to 35 mg/L), and 11 mg/L (range, 5 to 25 mg/L), in groups 1, 2, and 3, respectively (P = 0.006). The time to peak levels were 3.5 hours (range, 2.5 to 6.5 hours), 2.5 hours (range, 2.5 to 4.5 hours), and 4.5 hours (range, 2.5 to 12 hours), respectively (P = 0.2). TTA is safe and well tolerated. PMID- 18427287 TI - Medical therapy of reflux laryngitis. AB - Laryngeal signs and symptoms are frequently associated with gastroesophageal reflux disease (GERD). Establishing the diagnosis of laryngopharyngeal reflux (LPR), however, is enigmatic as there are no tests that specifically define GERD related laryngitis. Furthermore, in contrast to typical GERD, the treatment data for LPR using acid suppression with proton pump inhibitors has not shown a statistically significant advantage over placebo. This review highlights the current challenges for establishing the diagnosis of GERD-related LPR and focuses on the limitations of medical therapy directed toward gastric acid suppression. PMID- 18427286 TI - Differential effects of epsilon-aminocaproic acid and aprotinin on matrix metalloproteinase release in patients following cardiopulmonary bypass. AB - This study examined whether differential effects of 2 agents commonly used for hemostatic purposes during cardiac surgery, aprotinin and epsilon-aminocaproic acid (EACA), exist with respect to elevations in proinflammatory interleukins (ILs) and matrix metalloproteinases (MMPs) in patients undergoing coronary artery bypass surgery. Sixty patients were prospectively randomized to receive either aprotinin (1 x 10 KIU; n = 30) or EACA (5 g IV; n = 30), and blood samples were obtained for IL and MMP levels just before induction of anesthesia (Baseline), 10 minutes after separation from cardiopulmonary bypass (Post), and 6 hours after surgery (6 hours). IL-6 was increased at Post in the EACA group and increased further at 6 hours. In the aprotinin group, IL-6 was significantly increased only at 6 hours. MMP subtypes associated with inflammation, MMP-8, and MMP-9 were increased in the EACA group at Post and remained elevated at 6 hours. Thus, differential effects on IL and MMP release occurred between aprotinin and EACA, indicative of different mechanisms of action independent of hemostatic effects. PMID- 18427288 TI - Stem cells bioprocessing: an important milestone to move regenerative medicine research into the clinical arena. AB - Regenerative Medicine is a new, multidisciplinary field that combines expertise in biology, chemistry, engineering, materials, and medicine, to find solutions to some of the most challenging medical problems faced by humankind. Regenerative Medicine has the potential to impact the whole spectrum of health care, such as heart disease, emphysema, and diabetes. Regenerative Medicine employs various combinations of specially grown cells, tissues, and laboratory-made compounds to replace or amplify the body's natural healing process. The impact of Regenerative Medicine to the health care industry is likely to be comparable with that of antibiotics, vaccines and lately, monoclonal antibodies have had in clinical care. Regenerative Medicine is growing and maturing steadily; however, many challenges lie ahead. These include best cell source, most appropriate biomaterials, and reliable ways of expanding the cells and growing them in a three-dimensional environment (stem cell bioprocessing). This concise review deals with current achievements in the field, challenges that lie ahead and potential ways of having robust and reliable "off the shelf" cellular products. PMID- 18427289 TI - Stem cell and regenerative science applications in the development of bioengineering of renal tissue. AB - A rising number of patients with acute and chronic renal failure worldwide have created urgency for clinicians and investigators to search out alternative therapies other than chronic renal dialysis and/or organ transplantation. This review focuses on the recent achievements in this area, and discusses the various approaches in the development of bioengineering of renal tissue including recent discoveries in the field of regenerative medicine research and stem cells. A variety of stem cells, ranging from embryonic, bone marrow, endogenous, and amniotic fluid, have been investigated and may prove useful as novel alternatives for organ regeneration both in vitro and in vivo. Tissue engineering, developmental biology, and therapeutic cloning techniques have significantly contributed to our understanding of some of the molecular mechanisms involved in renal regeneration and have demonstrated that renal tissue can be generated de novo with similar physiologic functions as native tissue. Ultimately all of these emerging technologies may provide viable therapeutic options for regenerative medicine applications focused on the bioengineering of renal tissue for the future. PMID- 18427290 TI - Stem cells and tissue engineering applications of the genitourinary tract. AB - The field of regenerative medicine continues to make substantial advancements in therapeutic strategies addressing urologic diseases. Tissue engineering borrows principles from the fields of cell biology, materials science, transplantation and engineering in an effort to repair or replace damaged tissues. This review is intended to provide a current overview of the use of stem cells and tissue engineering technologies specifically in the treatment of genitourinary diseases. Current themes in the field include the use of adult stem cells seeded onto biocompatible resorbable matrices for implantation as tissue substitutes, which is conducive to host tissue in-growth. Injection therapy of adult stem cells for organ rehabilitation is also making strong headway toward the restoration of organ structure and function. With new data describing the molecular mechanisms for differentiation, work has begun on targeting tissues for regeneration by genetic modification methods. Promising laboratory discoveries portend the emergence of a new class of clinical therapies for regenerative medicine applications in the genitourinary tract. PMID- 18427291 TI - Tissue engineering craniofacial defects with adult stem cells? Are we ready yet? AB - Over three-quarters of all craniofacial defects observed in the US per year are cleft palates. Usually involving significant bony defects in both the hard palate and alveolar process of the maxilla, repair of these defects is typically performed surgically using autologous bone grafts taken from appropriate sites (i.e., iliac crest). However, surgical intervention is not without its complications. As such, the reconstructive surgeon has turned to the scientist and engineer for help. In this review, the application of the field of tissue engineering to craniofacial defects (e.g., cleft palates) is discussed. Specifically the use of adult stem cells, such as mesenchymal stem cells from bone marrow and Adipose-derived Stem Cells (ASCs) in combination with currently available biomaterials is presented in the context of healing craniofacial defects like the cleft palate. Finally, future directions with regards to the use of ASCs in craniofacial repair are discussed, including possible scaffold-driven and gene-driven approaches. PMID- 18427292 TI - Polymeric biomaterials in tissue engineering. AB - Polymeric biomaterials are one of the cornerstones of tissue engineering. A wide range of materials has been used. Approaches have shown increasing sophistication over recent years employing drug delivery functionality, micropatterning, microfluidics, and other technologies. Challenges such as producing three dimensional matrixes and rendering them deliverable through minimally invasive techniques have been addressed. A major recent development is the design of biomaterials for tissue engineering matrices to achieve specific biologic effects on cells, and vice versa. Much remains to be achieved, particularly in integrating other new technologies into the field. PMID- 18427293 TI - Collagen tissue engineering: development of novel biomaterials and applications. AB - Scientific investigations involving collagen have inspired tissue engineering and design of biomaterials since collagen fibrils and their networks primarily regulate and define most tissues. The collagen networks form a highly organized, three-dimensional architecture to entrap other ingredients. Biomaterials are expected to function as cell scaffolds to replace native collagen-based extracellular matrix. The composition and properties of biomaterials used as scaffold for tissue engineering significantly affect the regeneration of neo tissues and influence the conditions of collagen engineering. The complex scenario of collagen characteristics, types, fibril arrangement, and collagen structure-related functions (in a variety of connective tissues including bone, cartilage, tendon, skin and cornea) are addressed in this review. Discussion will focus on nanofibrillar assemblies and artificial synthetic peptides that mimic either the fibrillar structure or the elemental components of type I collagen as illustrated by their preliminary applications in tissue engineering. Conventional biomaterials used as scaffolds in engineering collagen-containing tissues are also discussed. The design of novel biomaterials and application of conventional biomaterials will facilitate development of additional novel tissue engineering bioproducts by refining the currently available techniques. The field of tissue engineering will ultimately be advanced by increasing control of collagen in native tissue and by continual manipulation of biomaterials. PMID- 18427294 TI - Biomaterial scaffolds in pediatric tissue engineering. AB - This article reviews recent developments and major issues in the use and design of biomaterials for use as scaffolds in pediatric tissue engineering. A brief history of tissue engineering and the limitations of current tissue-engineering research with respect to pediatric patients have been introduced. An overview of the characteristics of an ideal tissue-engineering scaffold for pediatric applications has been presented, including a description of the different types of scaffolds. Applications of scaffolds materials have been highlighted in the fields of drug delivery, bone, cardiovascular, and skin tissue engineering with respect to the pediatric population. This review highlights biomaterials as scaffolds as an alternative treatment method for pediatric surgeries due to the ability to create a functional cell-scaffold environment. PMID- 18427295 TI - Memory encoded throughout our bodies: molecular and cellular basis of tissue regeneration. AB - When a sheep loses its tail, it cannot regenerate it in the manner of lizards. On the other hand, it is possible to clone mammals from somatic cells, showing that a complete developmental program is intact in a wounded sheep's tail the same way it is in a lizard. Thus, there is a requirement for more than only the presence of the entire genetic code in somatic cells for regenerative abilities. Thoughts like this have motivated us to assemble more than just a factographic synopsis on tissue regeneration. As a model, we review skin wound healing in chronological order, and when possible, we use that overview as a framework to point out possible mechanisms of how damaged tissues can restore their original structure. This article postulates the existence of tissue structural memory as a complex distributed homeostatic mechanism. We support such an idea by referring to an extremely fragmented literature base, trying to synthesize a broad picture of important principles of how tissues and organs may store information about their own structure for the purposes of regeneration. Selected developmental, surgical, and tissue engineering aspects are presented and discussed in the light of recent findings in the field. When a sheep loses its tail, it cannot regenerate it in the manner of lizards. On the other hand, it is possible to clone mammals from somatic cells, showing that a complete developmental program is intact in a wounded sheep's tail the same way it is in a lizard. Thus, there is a requirement for more than only the presence of the entire genetic code in somatic cells for regenerative abilities. Thoughts like this have motivated us to assemble more than just a factographic synopsis on tissue regeneration. As a model, we review skin wound healing in chronological order, and when possible, we use that overview as a framework to point out possible mechanisms of how damaged tissues can restore their original structure. This article postulates the existence of tissue structural memory as a complex distributed homeostatic mechanism. We support such an idea by referring to an extremely fragmented literature base, trying to synthesize a broad picture of important principles of how tissues and organs may store information about their own structure for the purposes of regeneration. Selected developmental, surgical, and tissue engineering aspects are presented and discussed in the light of recent findings in the field. PMID- 18427296 TI - Bioengineering approaches to controlled protein delivery. AB - Proteins are of crucial importance in all biologic organisms, in terms of both structure and function. Their deficits play central roles in many pathologic states, and their potential as powerful therapeutic agents has been widely recognized. Many issues, however, exist in delivery of biologically active proteins to target tissues and organs. Recent advances in biomedical engineering have lead to development of advanced techniques for controlled delivery of peptides and proteins, paving the way for their efficient use in treating human injury and disease. With a particular emphasis on most recent advances, this review discusses currently available techniques for controlled delivery of proteins and considers future research directions. PMID- 18427297 TI - Tissue engineering and organ structure: a vascularized approach to liver and lung. AB - Over the past two decades, great strides have been made in the field of tissue engineering. Many of the initial attempts to develop an engineered tissue construct were based on the concept of seeding cells onto an avascular scaffold. Using advanced manufacturing technologies, the creation of a preformed vascular scaffold has become a reality. This article discusses some of the issues surrounding the development of such a vascular scaffold. We then examine of the challenges associated with applying this scaffold technology to two vital organ constructs: liver and lung. PMID- 18427298 TI - Bioengineering cartilage growth, maturation, and form. AB - Cartilage of articular joints grows and matures to achieve characteristic sizes, forms, and functional properties. Through these processes, the tissue not only serves as a template for bone growth but also yields mature articular cartilage providing joints with a low-friction, wear-resistant bearing material. The study of cartilage growth and maturation is a focus of both cartilage biologists and bioengineers with one goal of trying to create biologic tissue substitutes for the repair of damaged joints. Experimental approaches both in vivo and in vitro are being used to better understand the mechanisms and regulation of growth and maturation processes. This knowledge may facilitate the controlled manipulation of cartilage size, shape, and maturity to meet the criteria needed for successful clinical applications. Mathematical models are also useful tools for quantitatively describing the dynamically changing composition, structure and function of cartilage during growth and maturation and may aid the development of tissue engineering solutions. Recent advances in methods of cartilage formation and culture which control the size, shape, and maturity of these tissues are numerous and provide contrast to the physiologic development of cartilage. PMID- 18427299 TI - The development of a tissue-engineered cornea: biomaterials and culture methods. AB - The field of corneal tissue engineering has made many strides in recent years. The challenges of engineering a biocompatible, mechanically stable, and optically transparent tissue are significant. To overcome these challenges, researchers have adopted two basic approaches: cell-based strategies for manipulating cells to create their own extracellular matrix, and scaffold-based strategies for providing strong and transparent matrices upon which to grow cells. Both strategies have met with some degree of success. In addition, recent advances have been made in innervating a tissue-engineered construct. Future work will need to focus on further improving mechanical stability of engineered constructs as well as improving the host response to implantation. PMID- 18427300 TI - Tissue engineering in cleft palate and other congenital malformations. AB - Contributions from multidisciplinary investigations have focused attention on the potential of tissue engineering to yield novel therapeutics. Congenital malformations, including cleft palate, craniosynostosis, and craniofacial skeletal hypoplasias represent excellent targets for the implementation of tissue engineering applications secondary to the technically challenging nature and inherent inadequacies of current reconstructive interventions. Apropos to the search for answers to these clinical conundrums, studies have focused on elucidating the molecular signals driving the biologic activity of the aforementioned maladies. These investigations have highlighted multiple signaling pathways, including Wnt, fibroblast growth factor, transforming growth factor beta, and bone morphogenetic proteins, that have been found to play critical roles in guided tissue development. Furthermore, a comprehensive knowledge of these pathways will be of utmost importance to the optimization of future cell based tissue engineering strategies. The scope of this review encompasses a discussion of the molecular biology involved in the development of cleft palate and craniosynostosis. In addition, we include a discussion of craniofacial distraction osteogenesis and how its applied forces influence cell signaling to guide endogenous bone regeneration. Finally, this review discusses the future role of cell-based tissue engineering in the treatment of congenital malformations. PMID- 18427301 TI - Smart materials applications for pediatric cardiovascular devices. AB - "Smart Materials" are materials that change their shape, color, or size in response to an externally applied stimulus. While smart materials have already made a tremendous impact on our lives through their applications in liquid crystal displays, headphones, fuel injection systems, flexible cell phone antennas, and many other commercial products, they also have the potential to help many pediatric patients. This review focuses on with the present and potential applications of shape memory alloys, piezoelectric materials, and the relatively newer class of materials called magnetostrictive and ferromagnetic shape memory alloys in the design of pediatric cardiovascular devices. PMID- 18427302 TI - The development of tissue-engineered grafts for reconstructive cardiothoracic surgical applications. AB - Surgical correction of congenital heart defects often requires the use of valves, patches, or conduits to establish anatomic continuity. Homografts, xenografts, or mechanical prosthetic devices are frequently implanted during these surgical procedures. These grafts however lack growth potential, are associated with increased risk of thrombosis and infection and have limited durability, thus increasing the morbidity and mortality of their application in pediatric cardiac surgery. These limitations are being addressed through the development of living, biologic tissue-engineered valves, patches, and conduits. Pilot studies and phase 1 clinical trials are currently underway to evaluate their feasibility, safety, and efficacy. The optimal scaffold, cell source, and conditioning parameters, however, still remain to be determined and are areas of active research. PMID- 18427303 TI - Bioengineered tissues for urogenital repair in children. AB - The most common congenital abnormalities involve the genitourinary system. These include hypospadias, in which the urethral opening develops in an improper position, and bladder exstrophy, in which the bladder develops on the outer surface of the abdomen. Children with these conditions will require immediate and multiple reconstructive surgeries. Currently, reconstruction may be performed with native nonurologic tissues (skin, gastrointestinal segments, or mucosa), homologous tissues from a donor (cadaver or living donor kidney), heterologous tissues or substances (bovine collagen), or artificial materials (silicone, polyurethane, teflon). However, these materials often lead to complications after reconstruction, either because the implanted tissue is rejected, or because inherently different functional parameters cause a mismatch in the system. For example, replacement of bladder tissue with gastrointestinal segments can be problematic due to the opposite ways in which these two tissues handle solutes urologic tissue normally excretes material, and gastrointestinal tissue generally absorbs the same materials. This mismatched state can lead to metabolic complications as well as infection and other issues. The replacement of lost or deficient urologic tissues with functionally equivalent ones would improve the outcome of reconstructive surgery in the genitourinary system. This goal may soon be attainable with the use of tissue engineering techniques. PMID- 18427304 TI - Microsystems for biomechanical measurements. AB - The use of microtechnology to make biomechanical measurements allows for the study of cellular and subcellular scale mechanical forces. Forces generated by cells are in the few nanoNewton to several microNewton range and can change spatially over subcellular size scales. Transducing forces at such small size and force scales is a challenging task. Methods of microfabrication developed in the integrated circuit industry have allowed researchers to build platforms with cellular and subcellular scale parts with which individual cells can interact. These parts act as transducers of stresses and forces generated by the cell during migration or in the maintenance of physical equilibrium. Due to the size and sensitivity of such devices, quantitative studies of single cell and even single molecule biomechanics have become possible. In this review we focus on two classes of cellular force transducers: silicon-based devices and soft-polymer platforms. We concentrate on the biomechanical discoveries made with these devices and less so on the engineering behind their development because this is covered in great detail elsewhere. PMID- 18427305 TI - Antiangiogenic strategies in medulloblastoma: reality or mystery. AB - Medulloblastoma is the most common malignant brain tumor of childhood. Surgery, radiation therapy, and chemotherapy successfully cure many patients, but survivors can suffer long-term toxicities affecting their neurocognitive and growth potential; furthermore, there is no curative therapy in up to 30% of cases, mainly because of our incomplete understanding of many of the underlying molecular and cellular processes. Angiogenesis is a hallmark of the progression of medulloblastoma and, over the last years, investigators have sought to develop effective and less toxic antiangiogenic strategies, including the inhibition or destruction of abnormal blood vessels using either antiangiogenic or vascular disrupting agents. However, the results are conflicting principally because of the complex biology of tumor vasculature and the irregular geometry of the vascular system in real space. In addition, current targets of antiangiogenic therapy, such as vascular endothelial growth factor (VEGF), are thought to be critical for both physiologic and pathologic angiogenesis, and clinical side effects of anti-VEGF therapy are beginning to emerge. We here review the state-of the-art concerning antiangiogenic targets for medulloblastoma treatment, and discuss the complexity of the vascular system that intrinsically limits the efficacy of current strategies. PMID- 18427306 TI - Implications of lumbar plexus anatomy for removal of total disc replacements through a posterior approach. AB - STUDY DESIGN: An anatomic study in which the lumbar plexuses of 14 embalmed cadavers were dissected bilaterally and measured using a posterior approach. OBJECTIVE: To determine the cephalocaudal (root-to-root) distances and the mediolateral (root-to-tether) distances within the lumbar plexus and determine the feasibility for removal of a lumbar total disc replacement (TDR) through these anatomic spaces using a posterior approach. SUMMARY OF BACKGROUND DATA: Currently, lumbar TDRs are implanted primarily through an anterior retroperitoneal or transperitoneal approach. However, revision surgeries through these approaches can be complicated by significant adhesions, with potential injuries to intra- and retroperitoneal contents. Advancements in accessing anterior column structures through a posterior lumbar extracavitary approach suggest that posterior removal of TDRs may be an alternative. Unlike the thoracic extracavitary approach in which ligation of the thoracic nerve rarely leaves significant morbidity, the lumbar extracavitary approach cannot rely on the analogous ligation of the lumbar root to achieve access. Therefore, feasibility of the lumbar extracavitary approach depends on the presence of sufficient anatomic space between the tethered nerves of the lumbar plexus. METHODS: Fourteen adult cadavers (5 M/9F) were dissected through a posterior approach to expose the lumbar plexus bilaterally. The root-to-root distances at levels L2-S1 and corresponding root-to-tether distances at levels L3-L5 were measured bilaterally. RESULTS: Root-to-root distance was smallest at the male L5-S1 interval (11.7 +/- standard deviations 4.1 mm). Root-to-tether distance was smallest at the female L5 (43.1 +/- standard deviations 8.4 mm). These plexus measurements compare favorably with the CHARITE TDR components, in which the thickest sliding core is 11.0 mm in height and the largest endplate is 42.0 mm in width. CONCLUSION: This anatomic study suggests that posterior TDR removal is possible in the lumbar spine without undue risk to the surrounding nervous structures. PMID- 18427307 TI - Brown-Sequard syndrome produced by C3-C4 cervical disc herniation: a case report and review of the literature. AB - STUDY DESIGN: The article presents a case in which Brown-Sequard syndrome resulted from a painless C3-C4 disc herniation. OBJECTIVE: To raise spinal surgeons' awareness of this unusual clinical problem. SUMMARY OF BACKGROUND DATA: Brown-Sequard syndrome involves ipsilateral loss of motor function combined with contralateral loss of pain and temperature sensation. Brown-Sequard syndrome is commonly seen in the setting of spinal trauma or an extramedullary spinal neoplasm, but rarely it can be caused by a herniated cervical disc. METHODS: A 46 year-old man presented with progressive numbness and weakness in the left arm, mild neck pain, and reduced temperature sensation on the right side of the body. There was weakness in left arm and leg and proximal right lower limb. Magnetic resonance imaging showed large C3-C4 disc herniation compressing the spinal cord at that level. Anterior cervical discectomy and fusion with iliac crest bone graft was performed. RESULTS: Follow-up showed complete resolution of the neck pain, normal sensory function, and complete recovery of motor power in the left upper and right lower limb. There was a slight residual weakness in the left leg. CONCLUSION: Brown-Sequard syndrome is rarely caused by a cervical disc herniation. This etiology may be underdiagnosed but has a more favorable outcome in those cases where rapid diagnosis is followed by spinal cord decompression. PMID- 18427308 TI - The paracondylar process: an unusual and treatable cause of posttraumatic headache. AB - STUDY DESIGN: Case report and clinical discussion. OBJECTIVE: To describe a case of persistent posttraumatic headache associated with the presence of a paracondylar process, and the result of surgical resection of this osseous anomaly. SUMMARY OF BACKGROUND DATA: The paracondylar process is one of a group of congenital anomalies of the craniovertebral junction. This heterogenic group is classified in the literature based on embryologic origin. In general, the paracondylar process has little or no clinical significance. There are some cases reporting symptoms related to the paracondylar process, especially after head trauma. Few report improvement after surgery. METHODS: We describe a case of a 21 year-old woman who was seen for severe persistent daily headache and neck pain after minor head- and neck injury. Clinical examination revealed tenderness and an abnormal structure in the right cervico-occipital region by palpation. A conventional radiograph and computed tomography scan of the craniovertebral junction revealed a paracondylar process on the right side of the skull. RESULTS: After resection of the paracondylar process, headache and cervical pain were substantially reduced. CONCLUSION: This case underlines the possible clinical significance of congenital anomalies of the craniovertebral junction. They can become symptomatic after head trauma, causing headache and/or cervical pain. Surgical extirpation can relieve these symptoms. PMID- 18427309 TI - Computer-assisted surgical planning and image-guided surgical navigation in refractory adult scoliosis surgery: case report and review of the literature. AB - STUDY DESIGN: Case report and literature review. OBJECTIVE: In this case report, we present the utility of computer-assisted surgical planning and image-guided surgical navigation in the planning and execution of a major osteotomy to correct severe kyphoscoliosis. SUMMARY OF BACKGROUND DATA: Computer-assisted surgical planning is useful to appreciate the three-dimensional nature of scoliotic deformities and allows for operative maneuvers to be simulated on a computer before their implementation in the operating room. Image-guided surgical navigation improves surgical accuracy and can help translate a virtual surgical plan to the operative setting. METHODS: We report the case of a 38-year-old woman with severe, congenital kyphoscoliosis refractory to many previous surgeries, who presents with moderate progressive myelopathy and severe pain attributable to a sharp angular deformity at T12. Three-dimensional computed tomography reconstruction and computer-assisted surgical planning were used to determine the optimal corrective osteotomy. The surgical plan was translated to the operating room where a posterior vertebrectomy and instrumented correction were executed with the aid of image-guided surgical navigation. RESULTS: The osteotomy was safely performed resulting in improved sagittal and coronal alignments, as well as, correction of the sharp kyphoscoliotic deformity at the thoracolumbar junction. At 6-month follow-up, the patient's myelopathy and pain had largely resolved and she expressed high satisfaction with the procedure. CONCLUSION: We advocate this novel application of virtual surgical planning and intraoperative surgical navigation to improve the safety and efficacy of complex spinal deformity corrections. PMID- 18427310 TI - Two cases of Pseudomonas aeruginosa epidural abscesses and cervical osteomyelitis after dental extractions. AB - STUDY DESIGN: Case report. OBJECTIVES: To report 2 unusual cases of Pseudomonas aeruginosa epidural abscesses and cervical osteomyelitis after routine dental extractions and to review relevant literature. SUMMARY OF BACKGROUND DATA: Pseudomonas aeruginosa is a rare cause of cervical osteomyelitis in patients after dental extractions. Only 1 prior case could be found in the literature. METHODS: The cases of an 18-year-old male and a 23-year-old female are presented. PubMed was used to search for relevant literature. RESULTS: Our 2 patients presented with excruciating neck pain within 24 hours of routine dental extractions and, by imaging were found to have cervical epidural abscesses and osteomyelitis. Both patients were taken to the operating room for drainage and corpectomy and treated with prolonged courses of intravenous antibiotics. When seen in follow up 3 months later, neither patient demonstrated any neurologic sequelae. CONCLUSION: Pseudomonas aeruginosa epidural abscesses and osteomyelitis of the cervical spine have only rarely been reported in healthy patients after dental extractions. To our knowledge, the 2 patients reported here are only the second 2 such cases reported in the literature. Unfortunately, as in prior cases, these 2 patients had a significant delay in diagnosis. Therefore, a strong suspicion must be maintained for all patients presenting with neck pain after a recent dental extraction and appropriate imaging must be obtained urgently. PMID- 18427312 TI - Full-endoscopic interlaminar and transforaminal lumbar discectomy versus conventional microsurgical technique: a prospective, randomized, controlled study. AB - STUDY DESIGN: Prospective, randomized, controlled study of patients with lumbar disc herniations, operated either in a full-endoscopic or microsurgical technique. OBJECTIVE: Comparison of results of lumbar discectomies in full endoscopic interlaminar and transforaminal technique with the conventional microsurgical technique. SUMMARY OF BACKGROUND DATA: Even with good results, conventional disc operations may result in subsequent damage due to trauma. Endoscopic techniques have become the standard in many areas because of the advantages they offer intraoperatively and after surgery. With the transforaminal and interlaminar techniques, 2 full-endoscopic procedures are available for lumbar disc operations. METHODS: One hundred seventy-eight patients with full endoscopic or microsurgical discectomy underwent follow-up for 2 years. In addition to general and specific parameters, the following measuring instruments were used: VAS, German version North American Spine Society Instrument, Oswestry Low-Back Pain Disability Questionnaire. RESULTS: After surgery 82% of the patients no longer had leg pain, and 14% had occasional pain. The clinical results were the same in both groups. The recurrence rate was 6.2% with no difference between the groups. The full-endoscopic techniques brought significant advantages in the following areas: back pain, rehabilitation, complications, and traumatization. CONCLUSION: The clinical results of the full-endoscopic technique are equal to those of the microsurgical technique. At the same time, there are advantages in the operation technique and reduced traumatization. With the surgical devices and the possibility of selecting an interlaminar or posterolateral to lateral transforaminal procedure, lumbar disc herniations outside and inside the spinal canal can be sufficiently removed using the full endoscopic technique, when taking the appropriate criteria into account. Full endoscopic surgery is a sufficient and safe supplementation and alternative to microsurgical procedures. PMID- 18427313 TI - Full-endoscopic cervical posterior foraminotomy for the operation of lateral disc herniations using 5.9-mm endoscopes: a prospective, randomized, controlled study. AB - STUDY DESIGN: Prospective, randomized, controlled study of patients with lateral cervical disc herniations, operated either in a full-endoscopic posterior or conventional microsurgical anterior technique. OBJECTIVE: Comparison of results of cervical discectomies in full-endoscopic posterior foraminotomy technique with the conventional microsurgical anterior decompression and fusion. SUMMARY OF BACKGROUND DATA: Anterior cervical decompression and fusion is the standard procedure for operation of cervical disc herniations with radicular arm pain. Mobility-preserving posterior foraminotomy is the most common alternative in the case of lateral localization of the pathology. Despite good clinical results, problems may arise due to traumatization of the access. Endoscopic techniques are considered standard in many areas, since they may offer advantages in surgical technique and rehabilitation. These days, all disc herniations of the lumbar spine can be operated in full-endoscopic technique. With the full-endoscopic posterior cervical foraminotomy a procedures is available for cervical disc operations. METHODS: One hundred and seventy-five patients with full-endoscopic posterior or microsurgical anterior cervical discectomy underwent follow-up for 2 years. In addition to general and specific parameters, the following measuring instruments were used: VAS, German version North American Spine Society Instrument, Hilibrand Criteria. RESULTS: After surgery 87.4% of the patients no longer had arm pain, and 9.2% had occasional pain. The clinical results were the same in both groups. There were no significant difference between the groups in the revision or complication rate. The full-endoscopic technique brought advantages in operation technique, preserving mobility, rehabilitation, and traumatization. CONCLUSION: The recorded results show that the full-endoscopic posterior foraminotomy is a sufficient and safe supplement and alternative to conventional procedures when the indication criteria are fulfilled. At the same time, it offers the advantages of a minimally invasive intervention. PMID- 18427314 TI - Reproducible long-term disc degeneration in a large animal model. AB - STUDY DESIGN: Twelve goats were chemically degenerated and the development of the degenerative signs was followed for 26 weeks to evaluate the progression of the induced degeneration. The results were also compared with a previous study to determine the reproducibility. OBJECTIVES: The purpose of this study was determine whether this Chondroitinase ABC (CABC) induced goat model is reproducible and to study the development of the degeneration in time up to 26 weeks. SUMMARY OF BACKGROUND DATA: Injecting CABC into goat intervertebral discs results in mild disc degeneration after 12 weeks. Spontaneous recovery or leveling off of the degeneration has been reported before and is relevant when the goat model is used in regeneration studies. Reproducibility of the induced degeneration is relevant as well. METHODS: Twelve goats were used in this study. The development of degeneration was studied after the injection of 0.25 U/mL CABC intradiscally. The development of degenerative signs was studied after 18 (n = 6) and 26 (n = 6) weeks by means of radiograph, magnetic resonance imaging, macroscopic analysis, and histology and biochemical evaluation. The induced degeneration was compared with the results from a previous study, in which degeneration was induced similarly and analysis was performed after 12 weeks. RESULTS: The severity of the degenerative signs was mild and was consequently present in all parameters analyzed. When compared with the results after 12 weeks, the degeneration was similar in the present study. Spontaneous recovery was not observed up to 26 weeks. CONCLUSION: The injection with CABC in the intervertebral disc reproducibly results in mild disc degeneration in the goat. These findings corroborate the goat model as a suitable large animal model to evaluate mild disc degeneration and potential new therapies. PMID- 18427315 TI - Static electromagnetic fields generated by corrosion currents inhibit human osteoblast differentiation. AB - STUDY DESIGN: Human osteoblast cultures were exposed to a very low intensity static magnetic fields (SMF) to investigate its effects on osteoblast growth and differentiation. OBJECTIVE: Analysis of the effects of periprosthetic SMF on the growth and differentiation of human osteoblast cell cultures in vitro. SUMMARY OF BACKGROUND DATA: The effects of pulsed electromagnetic fields (PEMF) on cell proliferation, especially in human osteoblast-like cells is well described, whereas few data are available on the effects of SMF on osteoblast cell culture. We previously demonstrated that the proliferation of human osteoblast cultures is reduced when cells are exposed to a continuous low intensity SMF comparable to the one that occurs around metal devices (Ti spinal implant) because of the generation of electric currents between the screw (Ti6Al4V) and the rod (Ti). METHODS: Primary osteoblastic cells were isolated from a human femoral head. Osteoblast cultures were exposed to SMF and alkaline phosphatase activity was evaluated in the osteoblast cell cultures at different time points. Reverse transcriptase-polymerase chain reaction (RT-PCR) was performed to evaluate mRNA expression levels of osteocalcin, Runx2, and collagen I genes. RESULTS: The SMF treated cells showed a progressive increase in the alkaline phosphatase activity which, however, remained always lower than the one observed in the control group at each observation time (72 hours, 7 and 14 days). RT-PCR demonstrated that Runx2 and collagen I mRNA were downregulated following SMF stimulation, whereas no change in osteocalcin mRNA was observed. CONCLUSION: Continuous low-intensity electromagnetic field comparable to the one that generates around metal devices because of the generation of corrosion currents inhibits osteoblasts differentiation pattern and might contribute at least in part to a decrease in periprosthetic bone formation occurring in vivo. PMID- 18427316 TI - Salvage of C2 pedicle and pars screws using the intralaminar technique: a biomechanical analysis. AB - STUDY DESIGN: Human cadaveric biomechanical analysis. OBJECTIVE: The purpose of this study is to evaluate the ability of using 1 of the remaining 2 methods of instrumenting C2, should the initial method fail. SUMMARY OF BACKGROUND DATA: Although 3 different methods of C2 fixation (pedicle, pars, and laminar screws) are possible, occasionally an attempt at screw insertion fails. In such cases, the surgeon needs a viable alternative to salvage/obtain fixation to obviate the need to instrument an additional motion segment. METHODS.: Eleven fresh-frozen cadaveric specimens (Occ-C4) were DEXA scanned for bone mineral density. On the left side, pedicle screws were first inserted, then pulled out. Then, "salvage" pars screws were inserted, then pulled-out, followed by laminar screws. On the right, a similar sequence was repeated, except that a pars screw was followed by a pedicle screw, then a laminar screw. All screws were placed by experienced cervical spine surgeons. Insertional torque (IT) was measured in Newton-meters (Nm). Tensile loading to failure was performed "in-line"with the screw axis at a rate of 0.25 mm/s using a MTS 858 MiniBionix II System with data recorded as peak pull-out strength (POS) in newtons "N". RESULTS: Pedicle screws generated statistically greater IT and POS than other techniques as the initial fixation type (P < 0.0001). Similar trends were observed with transpedicular fixation as a salvage procedure (P > 0.05). Laminar screws yielded consistently higher POS values than pars fixation when applied in a salvage scenario (POS range: LS = 146 707 N; PrS = 8-548 N); however, high standard deviation precluded statistical significance (P > 0.05). Significant predictive relationship was established between IT and POS for all screws using Pearson correlation coefficient and bivariate linear regression analysis (r = 0.75 and r = 0.511, respectively; P < 0.01). CONCLUSION: Our results suggest that pedicle screws provide the strongest fixation for both initial and salvage applications. If they should fail, lamina screws appear to provide stronger and more reproducible fixation than pars screws. PMID- 18427317 TI - Beliefs about the causes and consequences of pain in patients with chronic inflammatory or noninflammatory low back pain and in pain-free individuals. AB - STUDY DESIGN: Case control study including 2 groups of patients with low back pain (LBP, inflammatory and noninflammatory) and a pain-free community control group. OBJECTIVE: We explored whether pain beliefs differ between patients with chronic LBP attributed to inflammatory or noninflammatory medical diagnoses, and between patients with chronic LBP and pain-free controls. SUMMARY OF BACKGROUND DATA: Beliefs strongly influence patients' engagement in and response to treatments for chronic LBP. It is unclear, however, whether unhelpful beliefs held by patients with chronic LBP are predominantly associated with diagnosis, or with other aspects of the patient's pain experience. METHODS: Patients and controls completed the pain beliefs questionnaire addressing beliefs about the causes and consequences of pain. Patients also completed questionnaires addressing catastrophizing (Coping Strategies Questionnaire), physical disability and bodily pain (SF-36 Health Survey), and psychological distress (Spielberger State-Trait Anxiety Inventory Short Form and Cognitive Depression Index). Variance analysis and chi2 test were used as appropriate, adjusting for effects of covariates and multiple comparisons. Linear regression and logistic regression were used to adjust for confounding factors. RESULTS: Patients with noninflammatory LBP more strongly endorsed organic pain beliefs (e.g., that pain necessarily indicates damage), and catastrophizing (e.g., that the pain is never going to get better), than did patients with inflammatory LBP (P < 0.01). Patients with inflammatory LBP, in turn, more strongly endorsed organic pain beliefs than did pain-free controls (P < 0.05). Endorsement of organic pain beliefs was associated with catastrophizing. CONCLUSION: Organic pain beliefs are associated with increased catastrophizing in patients with chronic LBP, and addressing these beliefs may help patients to manage their pain and disability. Meanings attributed to inflammatory and noninflammatory diagnostic labels may contribute to the different pain beliefs held by different patient groups. PMID- 18427318 TI - Endoscopic transforaminal discectomy for recurrent lumbar disc herniation: a prospective, cohort evaluation of 262 consecutive cases. AB - STUDY DESIGN: A prospective, cohort evaluation of 262 consecutive patients who underwent transforaminal endoscopic excision for recurrent lumbar disc herniation, after previous discectomy. OBJECTIVE: To review complications and results of the endoscopic transforaminal discectomy (ETD) for recurrent herniated disc with a 2-year follow-up. SUMMARY OF BACKGROUND DATA: Recurrent herniation is a significant problem, as scar formation and progressive disc degeneration may lead to increased morbidity after traditional posterior reoperation. The studies published until now on recurrent disc herniation concern various operative techniques, mostly the lumbar microdiscectomy, which is still seen as the standard. The advantage of ETD could be that there is no need to go through the old scar tissue and the procedure can be performed in local anesthesia. The disadvantage may be a long learning curve for the surgeon. METHOD: Between January 1994 and November 2002, 262 patients with primarily radicular problems underwent an ETD for a recurrent herniated disc. Two hundred and thirty-eight of these patients (90.84%) completed our 2-year follow-up questionnaire. Initial surgery of 82 patients was performed in-house, 180 external. Average age was 46.4 years. The female/male ratio was 29/71%. RESULTS: At 2-year follow-up 85.71% of patients rated the result of the surgery as excellent or good. 9.66% reported a fair and 4.62% patients an unsatisfactory result. Average improvement of back pain of 5.71 points and 5.85 points of leg pain on the VAS scale (1-10). According to Mac Nab, 30.67% of the patients felt fully regenerated, 50% felt their functional capacity to be slightly restricted, 16.81% felt their functional capacity noticeably restricted, and 2.52% felt unimproved or worse. All patients participated in a 3-month follow-up to establish the perioperative complications. The overall complication rate was 10/262 (3.8%), including 3 nerve root irritations and 7 early recurrent herniations (<3 month). There was no case of infection or discitis. After 3 months and within 2 years, 4 patients have been treated for a recurrent herniated disc in our own center and 7 patients have been treated elsewhere, resulting in a recurrence rate 11/238 (4.62%). CONCLUSION: ETD for recurrent disc herniation seems to be an effective method with few complications and a high patient satisfaction. PMID- 18427319 TI - Posterior only pedicle screw instrumentation with intraoperative halo-femoral traction in the surgical treatment of severe scoliosis (>100 degrees). AB - STUDY DESIGN: Retrospective clinical study. OBJECTIVE: To report the results of surgical correction achieved by intraoperative halo-femoral traction and posterior only pedicle screw instrumentation in severe scoliosis (scoliosis greater than 100 degrees ). SUMMARY OF BACKGROUND DATA: Although previous reports show the effectiveness of preoperative halo traction in the treatment of severe spinal deformity, the intraoperative use of halo-femoral traction in conjunction with posterior pedicle screw instrumentation has never been reported in patients with severe spinal deformity. METHODS: A total of 15 consecutive patients with severe (>100 degrees) thoracic idiopathic scoliosis and/or kyphoscoliosis operated by using intraoperative halo-femoral traction and posterior only pedicle screw instrumentation were included in the study. Subjects were analyzed by age at date of examination, gender, major coronal curve magnitude, major compensatory coronal curve magnitude, major sagittal curve magnitude, shoulder imbalance, and preoperative vital capacity of the lungs. Halo-traction related complications and short- and long-term complications were noted in each case. RESULTS: The average age at the time of surgery was 17.8 years (range, 16-19). There were 4 males and 11 females. The average improvement was 51% in the major thoracic curve, 33% in the compensatory lumbar curve, and 53% in the major sagittal curve. The average follow-up was 56 (range, 24-96) months. Loss of correction averaged 4 degrees for major thoracic curves and 2 degrees for thoracic kyphosis based on measurements at the final follow-up date. CONCLUSION: The use of intraoperative halo-femoral traction together with the wide facet resection and posterior release gradually provide a good correction and balance maintained by pedicle screw instrumentation. Intraoperative halo-femoral traction not only elongates spinal column but also elongates the thoracic cavity improving the compromised pulmonary function. PMID- 18427320 TI - Dual growing rod technique followed for three to eleven years until final fusion: the effect of frequency of lengthening. AB - STUDY DESIGN: Retrospective case review of children completing dual growing rod treatment at our institutions. Patients had a minimum of 2 years follow-up. OBJECTIVE: To identify the factors influencing dual growing rod treatment outcome followed to final fusion. SUMMARY OF BACKGROUND DATA: Published reports on dual growing rod technique results for early onset scoliosis demonstrate it to be safe and effective in curve correction and maintenance as well as in allowing spinal growth. METHODS: Between 1990 and 2003, 13 patients with no previous surgery and noncongenital curves underwent final fusion. All had preoperative curve progression over 10 degrees after unsuccessful nonoperative treatment. There were 10 females and 3 males. Average age was 6.6 +/- 2.9 years at initial surgery. There were 3 idiopathic, 1 nonspine congenital anomaly, and 9 syndromic patients. Analysis included age at initial surgery and final fusion, number and frequency of lengthenings, and complications. Radiographic evaluation included changes in Cobb angle, T1-S1 length, and instrumentation length over the treatment period. RESULTS: Cobb angle improved from 81.0 +/- 23 degrees to 35.8 +/- 15 degrees postinitial and 27.7 +/- 17 degrees after final fusion. Average number of lengthenings was 5.2 +/- 3 at an interval of 9.4 +/- 5 months. T1-S1 length increased from 24.4 +/- 3.4 to 29.3 +/- 3.6 cm postinitial and 35.0 +/- 3.7 cm postfinal fusion. Average growth was 1.46 +/- 0.66 cm/year. Those lengthened at or=30). Early follicular phase blood draw, anthropometric measurements, and a transvaginal ultrasonography were performed. Outcome measures were serum antimullerian hormone, inhibin B, estradiol, follicle-stimulating hormone, ultrasound ovarian volume, and antral follicle count. RESULTS: Mean antral follicle count was 7.6 for normal weight and 6.3 for obese women (P = 0.35). Proportions of normal weight (17%) versus obese women (22%) with antral follicle count less than 4 were similar. Ovarian volumes did not differ by body size. In adjusted models, antimullerian hormone levels in obese women were 77% lower on average than those in normal weight women (P = 0.02). Inhibin B levels were 24% lower in obese women compared with normal weight women (P = 0.08). Follicle stimulating hormone and estradiol were not associated with body mass index. CONCLUSIONS: Although antral follicle count did not differ by body size, antimullerian hormone was lower in obese compared with normal weight late reproductive age women. These data suggest that lower antimullerian hormone levels in obese late reproductive age women result from physiologic processes other than decreased ovarian reserve. PMID- 18427359 TI - Shaping outcomes using two Ni-Ti rotary instruments in simulated canals. AB - AIM: The aim of this study was to compare the shaping ability of Mtwo and ProTaper instruments in simulated curved root canals in resin blocks. METHODS: The study was carried out using 40 simulated canals in resin blocks, all of which had the same canal diameter (0.70 mm>D>0.20 mm), length (16 mm), and angle of curvature (40 degrees). These 40 canals were divided into 2 groups of 20 specimens. Both before and after instrumentation, all the resin specimens were photographed, and all the canals examined were measured at 4 different points of reference. Using a computer image analysis program, the photographs of corresponding sections were superimposed on one another. In this way we sought to evaluate the shaping ability of these 2 systems in relation to specific portions of the root canal, also with regard to maintenance of original canal curvature. The results were analysed using Student's t-test. RESULTS: Both rotary systems were found to respect the original root canal curvature, particularly in the areas most at risk of modification, namely the outer aspect of the curve in the apical third of the canal. The Mtwo instruments caused less transportation of the apex than the ProTaper instruments, but this difference was not statistically significant (P>0.05). CONCLUSION: According to the results of this study, both these instruments respected the original canal curvature, particularly in the areas at most risk of modification, and they also showed good shaping ability in curved canals, with minimum transportation of the apex. PMID- 18427358 TI - Plasma lipid-dependent and -independent effects of dietary soy protein and social status on atherogenesis in premenopausal monkeys: implications for postmenopausal atherosclerosis burden. AB - OBJECTIVE: : Atherosclerosis developed during premenopausal years predicts postmenopausal atherosclerosis burden. The objective of this study was to determine the effects of dietary soy protein isolate (SPI) and social status on atherogenesis and arterial gene expression in a premenopausal monkey model. DESIGN: : Socially housed premenopausal cynomolgus macaques (n = 84) were fed an atherogenic diet deriving protein from casein/lactalbumin or SPI (containing 1.88 mg isoflavones/g). After 36 months of diet consumption, iliac artery biopsies were assessed for atherosclerosis and expression of mRNA transcripts related to inflammation, macrophage and T-cell content, and estrogen receptors (ERs). RESULTS: : SPI reduced plaque size (P < 0.05), total plasma cholesterol, non-high density lipoprotein cholesterol (HDLc), and the total plasma cholesterol/HDLc ratio (all P < 0.003), while increasing triglycerides (P < 0.006) and HDLc (P < 0.0001). Arterial mRNA for CD68 (P < 0.001), CD3 (P < 0.02), and CD4 (P < 0.001) and inflammatory markers monocyte chemotactic protein-1, intercellular adhesion molecule-1, and interleukin-6 (all P < 0.0001) were also lower in the group receiving SPI. For most outcomes, this effect remained even after adjustments for plaque size and plasma lipid concentrations. Arterial ER-alpha was inversely associated with atherosclerosis (P < 0.02) and increased with SPI (P < 0.001). Subordinate monkeys had lower ER-beta (P < 0.02) and higher interleukin-6 (P < 0.05) transcripts but did not differ from dominant monkeys in extent of atherosclerosis (P > 0.9). CONCLUSIONS: : Premenopausal consumption of SPI had plasma lipid-independent beneficial effects on the pathobiological processes involved in atherosclerotic plaque development, thus potentially establishing the basis for reduced postmenopausal complications. Dominant social status provided similar, albeit less extensive, benefits in risk markers. PMID- 18427360 TI - Correlation of the temporomandibular dysfunction classification index with cephalometric indicators of facial structure: a study on adult female subjects. AB - AIM: The aim of this study was to determine whether associations exist between temporomandibular dysfunction index (TMI) and craniofacial morphology. METHODS: The sample consisted of 30 symptomatic (mean age 28.3+/-6.7; range 24-39 years, TMI 0.48+/-0.21) and 30 asymptomatic women (mean age 27.4+/-9.2; range 22-42 years, TMI 0.1+/-0.05) seeking orthodontic treatment at Department of Dental Sciences ''G. Messina'', University of Palermo, who had routine lateral cephalograms. Symptoms and signs of temporomandibular joint dysfunction (TMD) were assessed by a standardized clinical examination and the TMI (a clinical measure of dysfunction used to evaluate the severity of TMD). Linear and angular cephalometric measurements were taken to evaluate skeletal and dental characteristics of the two groups. Unpaired t test was used to compare the symptomatic subjects with the control subjects. RESULTS: The sella-nasion supramentale (SNB) angle (P<0.038, indicating mandibular retrognathism relative to cranial base), the lower facial height and the palatal plane-mandibular plane angle (P<0.025 and P<0.037 respectively, indicating hypodivergent facial profile) were significantly smaller in the symptomatic than in the asymptomatic women. The occlusal plane was steeper (P<0.033) and the cranial flexion was higher (P<0.035) in the symptomatic group than in the asymptomatic group. In addition the overjet (P<0.008) and the overbite (P<0.005) were significantly greater in symptomatic group than in the asymptomatic group. CONCLUSION: This study showed a significant correlation between dentofacial characteristics and TMD. PMID- 18427361 TI - Cemented-retained vs screw-retained implant restorations: an investigation on 1939 dental implants. AB - AIM: The aim of this work was to present a comparison of screw-retained and cemented-retained implant prostheses based on a retrospective analysis of 1 939 dental implants. The advantages, disadvantages, and limitations of those two different types of dental restorations are discussed and compared with the recent international literature. It is important to underline the influence of the attachment mechanism on many clinical aspects of implant dentistry. METHODS: A group of 527 patients was analyzed. All of the patients required implant rehabilitation. 1 939 dental implants were positioned between January 1997 and December 2003. RESULTS: The analyzed clinical trials show the presence of several factors essential to the long-term success of any implant prosthesis. Aesthetic, retrievability, passivity of framework, occlusion, ease of fabrication and cost and retention. In this study, 762 cemented dental prostheses were used for 1 551 dental implants placed. The survival rate of this rehabilitation was 98.4 % with an average of 34.7 month follow-up; 84 screwed-dental prostheses were placed on 330 dental implants. The success percentage of this type of prostheses was 100% with an average of 44.3 month follow-up. CONCLUSION: Cemented-implant prosthesis and screw-retained implant prosthesis analyzed in this investigation, present with advantages and disadvantages. Clinicians should be aware of the limitations and disadvantages of each type of prosthesis, so as to select the one that is most appropriate for a given clinical situation. PMID- 18427362 TI - Randomized clinical trial on the effects of full mouth disinfection versus conventional quadrant therapy in the control of chronic periodontitis. AB - AIM: This clinical trial investigates the effectiveness of full-mouth disinfection (FMD) versus conventional etiological therapy in patients with chronic periodontitis (CP). METHODS: The therapy effectiveness was assessed by a randomized trial, performed over 20 adult periodontitis (AP) patients, divided into two groups. Patients were recruited undergoing strict inclusion/exclusion criteria. The following parameters were considered to evaluate and compare the two procedures: bleeding on probing (BOP), Plaque Index (PLI), probing depth (PD), clinical attachment level (CAL). These clinical data were collected at baseline and at three follow-ups (three months, six months and twelve months from baseline). Each parameter was averaged within each group; then statistic comparisons were performed within groups and between groups. RESULTS: In the test group statistically significant improvements (P<0.001) were found for all parameters between baseline and every following review. The same result was reported in the control group (with a further significant difference between first and second review). Finally, the comparison between groups did not show any difference at any time for every parameter considered. CONCLUSION: FMD outcomes are similar to those of the conventional therapy and improvements can be achieved more quickly. FMD does not cause remarkable side effects and reduces the number of therapy sessions. Some aspects about this treatment need further research: maybe FMD could give an extra reduction of bacterial load, in comparison with traditional therapy, resulting in a longer free-infection period; that could allow a decrease in the frequency of supporting periodontal treatment. PMID- 18427363 TI - Osteo-promoting activity of OSTEOPLANT ANGIOSTAD in vitro. AB - AIM: There is an increasing need for an appropriate and readily-available material to reconstruct large bone defects, one of the most significant problems in the dental and maxillo-facial fields. The in vitro study examines the effects of OSTEOPLANT ANGIOSTAD, a product developed to increase osteoinductivity. METHODS: The product's biological properties were assessed by examining: the viability of cultured bone-marrow mesenchymal stem cells (MSC) through the methylthiazol tetrazolium assay; transforming growth factor (TGF)-b release by these cells through the enzyme-linked immunosorbent assay (ELISA) and the migration capacity of MSC and endothelial cells, by the in vitro wound closure test and transwell-migration assay, respectively. RESULTS: OSTEOPLANT ANGIOSTAD preserved MSC's viability and improved their capacity to release TGF-b1. It also increased in vitro wound healing by MSC and migration of endothelial cells. CONCLUSION: The results show that, since it increases the production by MSC of proangiogenic factors such as TGF-beta and promotes endothelial cell migration, OSTEOPLANT ANGIOSTAD may be an appropriate adjunct to accelerate the osteointegration of bone substitutes. PMID- 18427364 TI - Necrotizing sialometaplasia of the palate in a cocaine-using patient. A case report. AB - Cocaine abuse and the consequent prevalence of the damage associated with it have increased in the global population. The damage depends on the route of administration, where an important sequel of snorting is perforation of the nasal septum and palate. Necrosis of the palate, meanwhile, is also the manifestation of other diseases such as tertiary syphilis, necrotizing sialometaplasia and malignant neoplasms. The differential diagnosis of such lesions is important for determining appropriate treatment. A case of an ulcerated lesion in the palate of a cocaine-using patient whose diagnosis was necrotizing sialometaplasia is reported. PMID- 18427365 TI - An unusual oronasal fistula induced by prolonged cocaine snort. Case report and literature review. AB - In the last 50 years cocaine usage has had a growing incidence worldwide. Chronic nasal abuse of this drug can cause extensive oromaxillofacial destructive process as oronasal perforation. To author's knowledge the only type of oronasal fistula in cocaine abusers that has been reported to date has been perforation of the hard palate; a case of oronasal fistulas involving another region of oral cavity, the anterior gingival sulcus, has been reported only once. In this article authors present the first case of oronasal fistula involving hard palate and the right supero-lateral vestible induced by prolonged cocaine snort described in literature, review about the better surgical management for this lesion and the relationship between chronic cocaine abuse and elevated serum levels of c-ANCA. PMID- 18427366 TI - Oral laser surgical pathology: a preliminary study on the clinical advantages of diode laser and on the histopathological features of specimens evaluated by conventional and confocal laser scanning microscopy. AB - AIM: The effects of diode laser therapy reported in literature are both clinical and biological. The aim of the present preliminary study was to verify the clinical benefits such as reduction of postoperative pain, inflammatory response and bleeding and also the benefits concerning wound healing and histopathological alterations of specimens related to diode laser surgery. METHODS: The study was carried out on a group of 25 patients; we evaluated histologically the surgical specimens both by conventional optical microscopy and by autofluorescence by confocal laser scanner microscopy, paying attention to the incision margins and to the eventual presence of thermally induced changes/ damages. Both benign and premalignant or malignant lesions were treated with diode laser, with an output ranging from 2 to 7 Watt. RESULTS: Clinically we had several advantages (haemostasis, reduction of pain, healing without suture) and wound healing was always complete in 20-30 days. In the specimens evaluated histologically we noticed a good precision of surgical margins while changes induced by laser such as coagulation of proteins were present only with high power density output. Considering that nowadays the use of diode laser in the treatment of premalignant or malignant lesions is still debated, in this preliminary study no difficulty occurred with the observation of the specimens and no alterations were found. CONCLUSION: Thanks to the preliminary results obtained with this study, we encourage the use of diode laser also for malignant lesions, considering that an important factor to obtain specimens without alterations is the choice of the laser setting, strictly related to clinical experience, clinical situation, site and biological nature of the lesion. PMID- 18427367 TI - Mechanical evaluation of new injection needles for dental anesthesia. AB - AIM: In this study Authors want to evaluate the static and dynamic flexural behaviour of innovative injection needles for dental anaesthesia comparing to the traditional needles. METHODS: Four kinds of innovative injection needles (Carpule Free Flow-Heraeus Kulzer), different for length and external diameter, have been evaluated in comparison with the traditional ones (Carpule-Heraeus Kulzer), dimensions being equal except for the internal diameter. Static stiffness tests (10 for each kind of needles, resulting in a totality of 80 tests) and fatigue bending test (10 for each kind of needles, each tested at 3 different bending angles, resulting in a totality of 240 tests) have been conducted following the ISO 9626 norms. The final comparison among the obtained data have been conducted using a one way statistical analysis of variance (ANOVA). RESULTS: After stiffness tests the Carpule Free Flow injection needles and the traditional ones showed the same static behaviour, resulting in statistical comparable values of mean deflection at the maximum load allowed by ISO norms tests, whereas the 0.4 mm external diameter showed a statistically significant difference because the Carpule Free Flow needles achieved lower deflection values. After dynamic bending tests at 20 degrees both needle types showed the same behaviour, whereas at 30 degrees bending the Carpule Free Flow injection needles showed a higher number of breaks comparing to normal type, up to a maximum value of breaks (10 out of 10) for the 0.4 mm external diameter needles. After 50 degrees fatigue bending tests the Carpule Free Flow needles came to the break point after lower number of cycles than the Carpule needles. CONCLUSION: This study pointed out that, among anaesthesia injection needles, the Carpule Free Flow needles not only showed better clinical characteristics but also a mechanical behaviour, both static and dynamic, statistically similar to traditional ones, according to international standards. After fatigue bending higher than 20 degrees, exclusively made in this research in order to simulate critical or extreme conditions, all analysed Carpule Free Flow needles have been broken after a lower number of cycles. It is concluded that it is recommended not to repeatedly bend this kind of needles at more than 90 degrees. At the end of this study it may be suggested that this kind of needles cannot be repeatedly bended at angles more than 90 degrees. PMID- 18427368 TI - Evaluation of peri-implant tissues around single implant supporting single fixed crown: clinical considerations. AB - AIM: Assessment of peri-implant tissues around single implant-supported single fixed crown restorations can follow different protocols. The aim of this study was to asses peri-implant hard and soft tissues according to clinical periodontal parameters, modified for implant standard, and to radiographic parameters, and to test correlations between clinical parameters and historical bone loss. METHODS: Forty-two Branemarks implants (implant-supported single fixed crown restorations positioned between two natural teeth) inserted in 24 patients were assessed according to clinical and radiographic parameters, without removing the prosthetic crown. RESULTS: All implants were judged successful according to published criteria. Results of some clinical parameters differed between smokers and non-smokers. Radiographic measurements of peri-implant bone loss were in line with published data. Bone loss also correlated with the distance between adjacent teeth and implants. CONCLUSION: Radiographic parameters were found to be the most appropriate for evaluation of successful implants. While clinical parameters proved useful for distinguishing immediately between swollen and healthy tissues, they did not indicate a negative prognosis for osseointegration nor for potential complications affecting peri-implant soft tissues. PMID- 18427369 TI - Influence of storage solution and curing method on a microhybrid composite microhardness. AB - AIM: The purpose of this study is to evaluate and compare the Vickers microhardness of one microhybrid composite polymerized with different sources and stored in different solutions for up to 14 days. METHODS: Using a bipartite PTFE mould with 6 mm inner diameter and 3 mm high, 30 samples were manufactured with Charisma B1 shade for each polymerization procedures (halogen light, LED and halogen light and postcure cycles) stored in tree types of storage solution. RESULTS: The postcuring method tended to improve the microhardness, but was not statistically different from halogen or LED curing methods (P>0.05). The storage solutions interfered in surface hardness, with the samples eluted in red wine showing the lowest hardness values (P<0.05). After seven days, the hardness values were higher than the first day, but statistically equal to 14 days (P<0.05). CONCLUSION: On accordance with the findings of this study, different storage solutions can change the surface microhardness of a composite resin. An alcoholic solution seems most harmful to the composite. Samples postcured in autoclave had an improved mean value, however, without differing from those of the LED and halogen photo polymerized specimens. PMID- 18427370 TI - Failures in fabrication and treatment using ceramic crowns. AB - Dental ceramics is a constituent prosthodontic material with excellent aesthetic properties. It is stable, does not change shape, color or size, and is biotolerant. Treatment with ceramic restorations is a standard procedure in modern fixed prosthodontics. Because of its multiphase structure and the thermal processes it undergoes during the making of a fixed prosthodontic replacement, ceramics is highly sensitive to small failures in fabrication. These failures can develop into cracks that can ultimately cause fracture of restorations in use. This is why each phase of restoration in a dental practice and a dental laboratory requires professional skill. Selection of a definite ceramic system for a specific clinical case is more important than with other constituent materials. Patient selection, oral hygiene, masticatory force, as well as proper maintenance of instruments and devices for the fabrication of ceramic replacements are all important. PMID- 18427371 TI - Intermediate short-term tourniquet use during the preparation of a free vascularised fibula flap for mandibular reconstruction. A case report. AB - First described by Taylor et al. in 1975, the fibula flap is well established as a universal method for reconstruction of defects in several medical fields. Mostly a tourniquet is kept on during the whole procedure of harvesting the fibula flap. In some hospitals the operation is performed without tourniquet. The outcome is mostly described as successful, but functional impairment and donor site morbidity should not be neglected and severe complications are not frequently reported. In this article we describe a modification of the standard harvesting techniques to minimise the ischaemia time of the flap as well as the danger of severe blood loss. The tourniquet was only activated during the final disconnection of the fibular artery and was released immediately after the successful harvesting of the fibula flap. This method combines the safety of a tourniquet during the critical disconnection procedure and the advantages of a long perfusion of the donor site and the graft. PMID- 18427372 TI - Inflammatory myofibroblastic tumor of the alveolar mucosa of the mandible. AB - Inflammatory myofibroblastic tumor is a rare lesion composed of myofibroblastic spindle cells accompanied by inflammatory infiltrate. The objective of this paper is to report an uncommon case of inflammatory myofibroblastic tumor located in the alveolar mucosa of the mandible. A 33-year-old male presented an asymptomatic tumoral lesion, firm, pedunculated, pink-colored, covered by smooth mucosa, with focal ulceration, measuring 30x20x20 mm, located in the left posterior alveolar mucosa. Clinical diagnosis was soft tissue tumor. An excisional biopsy was made. Microscopic examination showed compact fascicular spindle cells proliferation with a diffuse inflammatory infiltrate of plasma cells, lymphocytes, and eosinophils. Large ganglion-like cells were observed. The lesional cells were immunopos-itive to vimentin, a-smooth muscle actin, muscle specific actin, and CD68. Negative immunostain was observed to S-100, Bcl-2, Ki-67, desmin, CD34, and cytokeratin. A diagnosis of inflammatory myofibroblastic tumor was performed. After 28 months of follow-up there was no recurrence. Although no evidence of oral inflammatory myofibroblastic tumor recurrence or malignant transformation has been reported, it has been observed that in inflammatory myofibroblastic tumor of other regions a prolonged follow-up is necessary after surgical excision. PMID- 18427373 TI - Successful conservative treatment of osteoradionecrosis of the mandible associated with pathological fracture. AB - RESULTS: Treatment of patients with osteoradionecrosis (ORN) of the mandible associated with pathologic fractures usually consists of a continuity resection of the mandible, leading to severe impairment of the patient's health and quality of life. Conservative treatment may be alternatively attempted; however, it is not usually employed. Here, we report a successful conservative treatment of ORN associated with pathologic fracture. A 48-year-old black man who had undergone radiotherapy for a squamous cell carcinoma in the tongue 6 years previously sought care complaining of facial swelling and draining episodes. Since the patient was fiercely resistant to any kind of hospital-based therapy, a conservative approach was proposed. Daily irrigations with 2% potassium iodate, followed by 3% oxygenated water, were performed by the patient at his house. In addition, local cleaning and debridement were weekly performed at our dental clinic. Systemic antibiotics were also administered. Throughout the therapy course, bone fragments were spontaneously exfoliated. Resolution of the ulcer and fracture was observed after 6 weeks of treatment. This report shows that ORN may be conservatively managed in specific situations, with good cosmetic and functional PMID- 18427374 TI - Multinodular neurilemmoma of the tongue: a case report with differential immunohistochemical profile. AB - Neurilemmoma is a distinctive, encapsulated, benign tumor, derived from Schwann cells. It is the most common neurogenic tumor, eventually occurring in the mouth, particularly the tongue. It occurs most often during the second or third decades of life, with predilection in females. Clinically, neurilemmomas present as a solitary circumscribed nodule, rarely being multinodular. We report a case of a 46 year-old female who presented a multinodular neurilemmoma of the tongue. She had a sessile nodule with two months of evolution. The presumptive diagnosis was of a benign mesenchymal neoplasm and an excisional biopsy was performed. During the excision it was found that the lesion was formed by two well-delimitated firm yellowish-white nodules. Histopathologic evaluation revealed an encapsulated benign tissue, formed by spindle cells forming irregular Antoni A and Antoni B patterns, not easily recognizable. Immunostaining was strongly positive for S-100 protein and negative for neurofilament. The lesion was excised and after 12 months of follow-up there was no recurrence. PMID- 18427376 TI - Enamel surface after debracketing of orthodontic brackets bonded with flowable orthodontic composite. A comparison with a traditional orthodontic composite resin. AB - AIM: Flowable composites showed acceptable shear bond strength for bonding orthodontic brackets, but it still lacks evidence if the routine clean-up of this type of composites from enamel surface after debracketing is susceptible of more difficult procedure, due to their fluidity, and, consequently, more enamel lesions, compared with traditional orthodontic composites. Thus, this in vitro study investigated this point. METHODS: Stainless steel brackets were bonded to 60 extracted human premolars using: 1) a new flowable composite (Denfil Flow) (group I; N=20); 2) a traditional flowable composite (Dyract Flow) (group II; N:20); 3) Transbond XT adhesive (group III; N=20). Twenty untreated teeth were the control group. After a routine debonding procedure, the adhesive remnant index (ARI) was calculated in each tooth. After the following clean-up procedure, enamel cracks (measurements about their frequency, their position in the tooth crown and their direction) were evaluated using fiber-optic transillumination. Finally, enamel surfaces were analyzed with scanning electron microscope to quantify enamel lesions RESULTS: A significantly higher frequency of enamel cracks, (from 65% to 80%), mostly in the cervical third of tooth crown (from 65% to 80%) and vertical (from 75% to 80%), were observed in the three study groups, compared with the control group (P<0.05), without difference among the three study groups. The index of enamel lesions was significantly higher in the three study groups, compared with control group (P<0.05), without significant differences among the three study groups. CONCLUSION: Flowable composites seem to show no relevant differences in terms of enamel lesions and cracks compared with traditional composites. PMID- 18427377 TI - In vitro retentive strength of metal superstructures cemented to solid abutments. AB - AIM: The aim of this study was to evaluate the effect of sandblasting on the retentive strength of metal single crowns luted with a resin cement to Straumann implant/abutment assemblies. METHODS: Fifty 4.1 mm-wide Straumann solid screw implants were mounted in self-polymerizing soft resin. Standard 5.5 mm-high, 8 degree tapered solid abutments were placed on each implant and torqued to 35 Ncm. Fifty metal castings were made using prefabricated burn-out caps. Each implant/abutment assembly and its corresponding metal casting was numbered and they were divided into two groups of 25. In the test group, the external surface of the abutments and the cavosurface of the corresponding metal casting were sandblasted. In the control group, neither the implant abutment nor the metal casting underwent sandblasting. Each metal casting was cemented onto its respective implant/abutment assembly using Panavia 21 (Kuraray Europe GmbH, Dusseldorf, Ger-many) resin cement. Specimens were then subjected to a pull-out test using a universal Instron testing machine. The load required to dislodge each crown was recorded and mean values were calculated for each group. Retention values were analyzed using the ANOVA test. RESULTS: The test group showed a higher mean retention value (83.78 kgf+/-19.61) than the control group (44.03 kgf+/-9.45) and the difference was statistically significant. CONCLUSION: Within the limitations of this in vitro study, the results suggest that sandblasting treatment significantly increases mechanical retention of crowns cemented using a resin cement. It is at the clinician's discretion to evaluate whether additional retention is desired in cementing an implant-supported fixed partial denture. PMID- 18427378 TI - Adhesive bond strength between a ceramic system and composite resin bonded by resinous cement submitted or not to thermocycling process. AB - AIM: The success of ceramic restorations is influenced by biocompatibility and resistance provided after bonding with an adhesive system. The aim of this in vitro study was to evaluate the adhesive bond strength between ceramic (IPS Classic V, Ivoclar Vivadent) and composite resin (Z-250, 3M), submitted or not to thermocycling treatment. METHODS: Forty ceramic cylinders were obtained with the following dimensions: 2 mm diameter and 3 mm high. The bond surface was polished with polishing paper (#600), conditioned with 10% hydrofluoric acid and silanized, according to manufacturer's recommendations. Later, the ceramic cylinders were bonded, with resinous cement (Rely X, 3M) to composite resin cylinders (Z-250, 3M) with the same dimensions, under 1 kg load. The assemblies were divided into two groups submitted to the following treatments: Group 1 (control): immersed in distilled water at 37 degrees C for 3 days, and Group 2: thermocycled (500 cycles) into two baths (5 degrees C and 55 degrees C), with 1 min of storing time each. In order to evaluate the adhesive resistance, the assemblies were tractioned in a universal testing machine (EMIC MEM 2000) with a crosshead speed of 0.5 mm/min. Obtained data were submitted to statistical analysis (ANOVA and Tukey's test). RESULTS: The means (MPa) and standard deviation obtained for Groups 1 and 2 were, respectively, 29.64+/-9.48 and 17.55+/-4.78. Group 1 presented significantly higher values than Group 2 (P<0.05). CONCLUSION: The results obtained in this study showed that the thermocycling procedure decreased the adhesive resistance between the materials studied. PMID- 18427379 TI - Salivary cortisol levels in young adults with temporomandibular disorders. AB - AIM: Causes of temporomandibular disorders (TMDs) are multifactorial, involving both physical and psychological factors. The aim of this study was to evaluate the salivary cortisol levels in young adults with TMDs. METHODS: The Research Diagnostic Criteria (RDC)/TMDs were used to establish the diagnosis of TMDs regarding the physical (axis I) and psychological factors (depression and somatization; axis II). Young adults of both genders composed the TMDs males (21.7+/-2.21 years, n=10) and females (22.7+/-1.95 years n=10) groups, and 20 healthy matched controls composed the control male (22.5+/-1.96 years n=10) and female (22.4+/-2.17years n=10) groups. Saliva samples were collected immediately on awakening and at bedtime. The cortisol levels were measured by enzyme immunoassay. RESULTS: The female group with TMDs showed a significantly higher cortisol level (2.890+/-0.411 mg/dL) than the corresponding control group in the morning evaluation (2.470+/- 0.486 mg/dL). The depression scores were significantly higher for the female group with TMDs (1.3+/-0.18) in comparison with the control female (0.72+/-0.18) and male groups (0.51+/-0.13). The groups with TMDs showed significantly higher somatization scores (0.69+/-0.16 for males and 1.38+/-0.23 for females) in comparison with the controls (0.23+/-0.10 for males and 0.78+/-0.15 for females), these also being significantly higher for the females than males. CONCLUSION: It was concluded that cortisol levels are higher in TMDs in females, who had elevated depression and somatization scores. The results infer that the evaluation of salivary cortisol could be a promising tool in the diagnosis of TMDs associated with psychological factors. PMID- 18427380 TI - Functional endoscopic sinus surgery for the treatment of Aspergillus mycetomas of the maxillary sinus. AB - AIM: The proper treatment for Aspergillus mycetoma (AM) of the maxillary sinus is a complete removal of the sinus fungal masses and improvement of aeration for the involved sinuses. We report our experience in the treatment of AM of the maxillary sinus by functional endoscopic sinus surgery (FESS). METHODS: Thirteen immunocompetent patients with AM underwent FESS under general anesthesia. Mycotic concretions were endonasally removed through the enlarged maxillary natural ostium. Care was taken to avoid any removal of the maxillary sinus mucosa. Only in one case complete removal could not be achieved and therefore we opened the maxillary sinus via the canine fossa and cleared it of fungal masses. Follow-up has ranged from 6 months to 4 years after surgery. Patients were all closely followed postoperatively with serial endoscopic examinations in order to verify the maintenance of opening of the maxillary natural ostium. RESULTS: Histological examination was positive for Aspergillus in all the patients. Postoperative radiographs and endoscopic examinations revealed maintenance of the antrostomy performed during FESS without mucosal degeneration and no evidence of recurrence. All patients were free of symptoms after a mean follow-up of 31 months. None of the patients required a second procedure. CONCLUSION: Surgical treatment of AM with FESS appears a reliable and safe surgical treatment. Sinus physiology is preserved in the event of a future bone reconstruction for prosthetic purposes. PMID- 18427381 TI - Modified surgical approach for removal of an ivory osteoma in the paranasal sinus. A case report. AB - Osteomas are benign tumors that consist mainly of mature compact or cancellous bone. The most common site in maxillofacial complex is the mandible, particularly the angle, followed by sinuses. The authors report a case of an ivory osteoma of the paranasal sinus in a 16 year-old male patient. The lesion was located in the left paranasal sinus (frontal, ethmoidal and maxillary), obstructing the nasolacrimal duct, resulting in dacryocystitis and frontal mucocele. The ethmoid and orbital portions were approached and excised through a Weber-Ferguson incision and maxillary osteotomy was carried out to remove the ivory osteoma, considering its location in the central and deep region of the face. After tumor removal and drainage of frontal and ethmoidal sinuses, the bone flap was repositioned using titanium microplates and screws. Soft tissue was then sutured. The patient was seen again at 3 weeks, 3 months and 2 years after surgery. Overall, he showed a good recovery and wound healing. A multispeciality team approach is advisable in such cases if radical excision is necessary. A craniofacial approach made radical single stage excision of this multicompartmental ivory osteoma possible with an uneventful postoperative period. PMID- 18427383 TI - The dream to re-build the human body. PMID- 18427382 TI - Treatment of submandibular fibromatosis with chemotherapy and surgery. A case report. AB - Fibromatoses are a broad group of fibrous tissue proliferation that arise from the musculoaponeurotic structures. Because of the locally aggressive behaviour and the high recurrence rate of these tumors, various treatment options are suggested but there is still no optimal therapy for these diseases. This paper describes a case of infantile fibromatosis of the submandibular region, which was successfully treated with an interdisciplinary approach using chemotherapy and surgery. A three-year-old girl with a growing submandibular mass over a period of six months is described. The tumor was clinically aggressive, causing bone erosion and invading adjacent soft tissue structures. The diagnosis of infantile fibromatosis was established on histological and immunohistochemical findings. With a combined treatment based on ''low-dose'' chemotherapy including vinblastine and methotrexate and surgery; complete remission was achieved in 30 months. Management of head and neck fibromatoses in child-hood needs a specific approach. Combined treatment by chemotherapy and nonmutilating surgery seems to offer the best potential for cure. PMID- 18427385 TI - Stem cell sources for cardiac regeneration. AB - Cell-based cardiac repair has the ambitious aim to replace the malfunctioning cardiac muscle developed after myocardial infarction, with new contractile cardiomyocytes and vessels. Different stem cell populations have been intensively studied in the last decade as a potential source of new cardiomyocytes to ameliorate the injured myocardium, compensate for the loss of ventricular mass and contractility and eventually restore cardiac function. An array of cell types has been explored in this respect, including skeletal muscle, bone marrow derived stem cells, embryonic stem cells (ESC) and more recently cardiac progenitor cells. The best-studied cell types are mouse and human ESC cells, which have undisputedly been demonstrated to differentiate into cardiomyocyte and vascular lineages and have been of great help to understand the differentiation process of pluripotent cells. However, due to their immunogenicity, risk of tumor development and the ethical challenge arising from their embryonic origin, they do not provide a suitable cell source for a regenerative therapy approach. A better option, overcoming ethical and allogenicity problems, seems to be provided by bone marrow derived cells and by the recently identified cardiac precursors. This report will overview current knowledge on these different cell types and their application in cardiac regeneration and address issues like implementation of delivery methods, including tissue engineering approaches that need to be developed alongside. PMID- 18427386 TI - Cardioprotection and cardiac regeneration by mesenchymal stem cells. AB - Mesenchymal stem cells (MSC) are promising candidates in the emerging field of regenerative medicine. MSC have been applied in numerous experimental and preclinical studies evaluating their therapeutical potential in various models of diseases. For cardiac applications, especially myocardial ischemia, MSC have been shown to provide an interesting therapeutical potential. However, to date the mechanisms of their beneficial effects on the cardiac tissue are only faintly elucidated. On the one hand we do not completely understand the biology of the MSC, on the other hand their interactions with the complex in vivo situation e. g. after myocardial infarction and the following remodeling processes are difficult to study. Initially, with respect to their in vitro differentiation capacity, it has been assumed that transplanted MSC may, promoted by the local microenvironment, differentiate in vivo into cells of cardiomyogenic phenotype or even cardiomyocytes, integrate themselves into the myocardium and help to regenerate the damaged tissue. The finding that after transplantation only a minor percentage of the MSC showed long term survival and persistent lack of evidence for in vivo differentiation make the original mechanistic concept of in vivo cardiomyogenic differentiation questionable. More recent studies suggest that the transplanted MSC may interact with the local tissue releasing paracrine factors and may hereby support the regenerative process. In this article, referring to experimental and preclinical studies, characteristics, sources, differentiation and paracrine activity of MSC contributing to cardioprotection and cardiac regeneration will be discussed. PMID- 18427384 TI - Targeting of cancer stem/progenitor cells plus stem cell-based therapies: the ultimate hope for treating and curing aggressive and recurrent cancers. AB - The rapid progression from aggressive primary cancers into locally advanced and invasive and/or metastatic diseases remains a big obstacle for an early diagnosis and curative therapeutic intervention for cancer patients. The late-stage leukemias and disseminated and metastatic sarcomas, melanomas, brain tumors and epithelial cancers are the devastating diseases associated with a high rate of recurrence after treatment with the conventional clinical therapies including surgery, ionizing radiation, hormonal therapy and systemic chemotherapy, which generally lead to the death of patients. Therefore, the establishment of the molecular events underlying cancer initiation and progression into locally invasive and metastatic diseases is of major interest in basic cancer research as well as for the development of new effective clinical therapeutic options against the recurrent and lethal cancers. Recent advances have led to the identification of specific oncogenic products that are implicated in the malignant transformation of adult stem/progenitor cells into leukemic or tumorigenic and migrating cancer stem/progenitor cells during cancer progression. Of therapeutic interest, the molecular targeting of deregulated signaling elements in cancer stem/progenitor cells and their local microenvironment represents a new potential strategy for the development of more effective clinical treatments against aggressive cancers. Particularly, the combined use of chemotherapeutic drugs to eradicate cancer-initiating cells with hematopoietic stem cell or genetically modified stem cell transplant is emerging as potential cancer treatments that hold great promise in the area of clinical cancer research. These targeting and stem cell-based therapies may offer the ultimate hope for treating and even curing the patients diagnosed with locally advanced cancers at high risk of recurrence, metastatic and/or relapsed cancers in the clinics. PMID- 18427388 TI - Neurogenesis and neurodegenerative diseases in human. AB - Acute and chronic neurodegenerative diseases are the most common neurological disorders in human and affect millions of individuals worldwide. While the specific clinical presentation varies among such diseases, their common feature is neural cell death. Yet, despite major advances in the understanding of neural cell death, effective treatment for these diseases remains one of the foremost challenges for medicine today. The transplantation of embryonic cells into the diseased brain in human has emerged out a mere theory and is possible as a practical application. This advancement, however, has raised important ethical, technical and immunological concerns. Studies have documented that neurogenesis occurs in the adult brain and that endogenous neural stem/progenitor cells (NSCs) respond to neurodegenerative diseases, suggesting that it might be possible for dead or injured neural cells to be replaced by endogenous NSCs. In this regard it is especially interesting to know the biological behaviors of endogenous NSCs in response to neurodegenerative diseases. Understanding the mechanisms underlying these changes could lead to the development of new strategies for treating neurological diseases using endogenous NSC pool. PMID- 18427387 TI - Stem cells and molecular strategies to restore hearing. AB - Hearing loss is a costly and growing problem for the elderly population worldwide with millions of people being affected. There are currently two prosthetic devices available to minimize problems associated with the two forms of hearing loss: hearing aids that amplify sound to overcome middle ear based conductive hearing loss and cochlear implants that restore some hearing after neurosensory hearing loss. The current presentation provides information on the treatment of neurosensory hearing loss. Although the cochlear implant solution for neurosensory hearing loss is technologically advanced; it still provides only moderate hearing capacity in neurosensory deaf individuals. Inducible stem cells and molecular therapies are appealing alternatives to the cochlear implant and may provide more than a new form of treatment as they hold the promise for a cure. To this end, current insights into inducible stem cells that may provide cells for seeding the cochlea with the hope of new hair cell formation are being reviewed. Alternatively, similar to induction of stem cells, cells of the flat epithelium that remains after hair cell loss could be induced to proliferate and differentiate into hair cells. In either of these strategies, hair cell specific genes known to be essential for hair cell differentiation or maintenance such as ATOH1, POU4F3, GFI1, and miRNA-183 will be utilized with the hope of completely restoring hearing to all patients with hearing loss. PMID- 18427389 TI - Tissue stem cells and cancer stem cells: potential implications for gastric cancer. AB - Gastric cancer remains the second leading cause of death in the world today, making the search for its molecular and cellular basis an important priority. Though recognition of the tight link between inflammation and tumorigenesis is centuries old, only recently are the pieces of the etiological puzzle beginning to fall together. Recent advances in gastric stem cell biology appear to be central to this slowly resolving puzzle. At least two types of stem cells may be important. Resident adult or tissue stem cells may, in a chronically inflamed environment, slowly acquire a series of genetic and epigenetic changes that lead to their emergence as ''cancer stem cells''. This scenario has not yet been proven experimentally, although the first step, prospective recognition of a gastric stem cell has recently been conquered. Alternatively, the setting of chronic inflammatory stress and injury may lead to loss of the indigenous gastric stem cells from their niches; bone marrow derived stem cells may then be recruited to and engraft into the gastric epithelium. Such recruited cells have the potential to contribute to the tumor mass. Indeed, evidence supporting this scenario has been published. Here, we review these recent findings and discuss implications for the future. PMID- 18427390 TI - Differentiation of embryonic and adult stem cells into insulin producing cells. AB - Replacement of insulin producing cells represents an almost ideal treatment for patients with diabetes mellitus type 1. Transplantation of pancreatic islets of Langerhans is successful in experienced centers. The wider application of this therapy, however, is limited by the lack of donor organs. Insulin producing cells generated from stem cells represent an attractive alternative. Stem cells with the potential to differentiate into insulin producing cells include embryonic stem cells (ESC) as well as adult stem cells from various tissues including the pancreas, liver, bone marrow and adipose tissue. The use of human ESC is hampered by ethical concerns but research with human ESC may help us to decipher important steps in the differentiation process in vitro since almost all information available on pancreas development are based on animal studies. The present review summarizes the current knowledge on the development of insulin producing cells from embryonic and adult stem cells with special emphasis on pancreatic, hepatic and human mesenchymal stem cells. PMID- 18427391 TI - Fetal alcohol syndrome (FAS): a not neglectable disease. PMID- 18427392 TI - Prevalence of hepatitis C virus/hepatitis G virus co-infection in Turin: updated data ten years later. PMID- 18427394 TI - Central European Vascular Forum. PMID- 18427393 TI - Kearns-Sayre syndrome with corneal dystrophy, hypomagnesemia, GH deficiency, and reduced visual acuity. PMID- 18427395 TI - Genetics of venous leg ulcer: an indication to priority of varicose veins treatment? PMID- 18427396 TI - Consensus document on intermittent claudication from the Central European Vascular Forum 1st edition - Abano Terme (Italy) - May 2005 2nd revision - Portroz (Slovenia) September 2007. PMID- 18427397 TI - Monocytes contribute to the atherosclerotic cap by transformation into fibrocytes. AB - AIM: The stability of an atherosclerotic plaque is a key-determining factor in the clinical outcome of cardiovascular disease. In this respect, smooth muscle (SM) alfa actin positive cells play an important role in maintaining plaque stability through formation of a fibrous cap. Recent evidence suggests that circulating progenitors may be a source of these cells. We hypothesized that they may be fibrocytes bone-marrow derived cells that acquire SM-like characteristics, including the expression of SM alfa actin. METHODS: We examined human carotid endarterectomy specimens for the presence of fibrocytes by immunohistochemistry staining for CD34/procollagen I and leukocyte specific protein-1/procollagen I) and examined fibrocyte differentiation in vitro. RESULTS: Fibrocytes were found in regions of plaque growth/healing. They possessed a SM-like spindle shape, produced collagen, and consistent with being fibrocytes they co-localized with transformation growth factor beta, but not serum amyloid P factors, known to promote and inhibit their formation, respectively. While fibrocytes were detected in regions of new growth in 35/40 specimens, only 1/3 of the specimens expressed the SM cell marker calponin, and smoothelin was absent, in these regions. CONCLUSION: Our results demonstrate that fibrocytes contribute to formation of the fibrous cap. With fibrocytes being a monocyte derived cell, we suggest that monocytes may play a more crucial role in the clinical outcome of atherosclerosis than previously realized as they not only contribute directly to plaque instability (through foam cell formation), but also promote plaque stability by transformation into a fibrocyte. PMID- 18427398 TI - Non-cardiac arterial disease in Catalonia: patient profiles and quality of clinical records by characteristics of the Vascular Surgery Unit. AB - AIM: Non-cardiac arterial disease (NCAD) is a frequent cause of hospital admission. The aim of this study was to investigate differences in patient profiles and clinical records as a function of the size of the Vascular Surgery Unit (VSU). METHODS: Retrospective observational study. Stratified cluster sampling and selection of patients hospitalized for NCAD. ANALYSIS: 1) description of patient profiles, quality of clinical records, and VSU [availability of diagnostic (DR) and therapeutic (TR) resources, and of written protocols (WP)]; 2) association between these variables and size of VSU. RESULTS: The sample consisted of 14 hospitals, 6 with a VSU of 15 or fewer beds (VSU < or = 15B) and 8 with >15 beds (VSU >15B). The mean number of DRs, TRs and WPs was 9, 2.8 and 2 in VSUs < or = 15B, and 11.5, 6.5 and 3.3 in VSUs >15B. The proportion of patients older than 70, female, with ischemic disease, or with coexisting diabetes was significantly higher in VSUs < or = 15B (67%, 31%, 95% and 57%, respectively) than in VSUs >15B (58%, 22%, 69% and 48%). Comorbid conditions and treatment during admission and at discharge were documented significantly less frequently in the clinical records in VSUs < or = 15B. Risk factors were under reported in the clinical records of both types of VSU. CONCLUSION: Patient profiles and the quality of clinical records vary by size of VSU. Under-reporting of risk factors may hinder the implementation of prevention and treatment measures. PMID- 18427399 TI - Men with atherosclerotic stenosis of the carotid artery have lower testosterone levels compared with controls. AB - AIM: There is evidence to suggest an inverse association between serum levels of testosterone and coronary heart disease. The aim of this study was to compare endogenous sex hormone levels of men with severe internal carotid artery (ICA) atherosclerosis with age-matched controls. METHODS: Metabolic parameters and sex hormones were measured or calculated in 124 male patients undergoing carotid endarterectomy for high grade ICA stenosis and in 124 age-matched male controls. The presence or absence of atherosclerotic stenosis of ICA was determined by high resolution B-mode ultrasound. RESULTS: The cases had statistically significant lower levels of total testosterone (TT) (medians: 3.8 microg/L versus 4.3 microg/L, P=0.005) and sex hormone binding globulin (SHBG) (means: 39.8+/-17.2 versus 54.3+/-34.3 nmol/L, P<0.001) compared to controls. Multivariate linear regression analysis, adjusted for all clinical and physiologic parameters, showed a significant inverse association between ICA stenosis and TT (b=-0.158, P=0.013) and SHBG (beta=-0.259, P<0.001). CONCLUSION: This study provides evidence of a positive association between low serum androgen levels and severe ICA atherosclerosis in men. It suggests that higher, but physiological, levels of androgens could have a protective role in the development of atherosclerosis. PMID- 18427400 TI - The prevalence of carotid artery stenosis in an unselected hospitalized stroke population. AB - AIM: The aim of this study was to describe the number and severity of carotid artery stenosis in an unselected stroke population in hospital. METHODS: The carotid arteries were investigated consecutively with color-coded duplex scanning in patients suspected of having stroke and admitted to a stroke unit during a 6 month period. Percent internal carotid artery stenosis by diameter reduction was described. RESULTS: A total of 144 patients were included in the investigation and the mean age was 75 years. The final diagnosis was stroke in 126 patients, while 18 had transient ischemic attacks. On the side, relevant to the neurologic deficit, a stenosis of >70% diameter reduction was observed in 4 patients and occlusion in 3. Severe stenosis and occlusion was found to have almost the same incidence on the contralateral side. Altogether 46 stenoses >30% (16.3%) were observed in 282 arteries investigated. The distribution was equal between the two sides. CONCLUSION: These findings indicate that few patients are eligible for surgery. However, routine duplex ultrasound examination in stroke patients gives information whether there are carotid arterial lesions, which could be a source of emboli. Such information can also be a guide for further medical treatment and lifestyle modification. PMID- 18427401 TI - The role of trans fatty acids in atherosclerosis, cardiovascular disease and infant development. AB - The process of partial hydrogenation converts vegetable oils in semisolid fats, like margarines, that contain high concentrations of trans fatty acids (TFA) and are commonly used in bakery, as well as for deep frying in fast food chains and other restaurants. Initially, these fats were considered the healthy solution, because they substituted butter and other cholesterol fats. However, in the last decades there has been continuing accumulation of evidence that TFA have potential harmful action in blood lipid metabolism, atherosclerosis development and cardiovascular disease, as well as in infant development. Consequently, many countries have enacted in order to reduce total TFA percentage in the daily fat intake, while others are waiting strongest evidence to enact. This article reviews the evidence of the effects of TFA, in relation to atherosclerosis, cardiovascular disease, inflammation and diabetes, and infant development. PMID- 18427402 TI - Acute thrombosis of abdominal aorta and hypercoagulable disorders. AB - Acute abdominal aortic occlusion (AAAO) is a rare, life threatening condition, which usually occurs in elderly patients, causing challenging management issues. In patients who have no cardiac or vascular disease this catastrophic event is very rare and is due to hypercoagulable disorders. This study reviews the literature on AAAO in hypercoagulable states in the light of our experience on a case of an acute thrombosis of nonaneurysmal, nonatherosclerotic abdominal aorta in a female patient with protein S deficiency and Sjogren's syndrome and her younger brother, which was found to have atherosclerotic involvement of distal aorta and elevation in homocysteine levels. Because of a misleading clinical presentation, the diagnosis was delayed and conservative treatment failed. Both were successfully treated with emergency aorto-bifemoral grafting. Other cases of arterial thrombosis and hypercoagulable disorders were found in first-degree relatives. Our experience and the review of the literature suggest that the interaction between host and environment factors can lead to acute thrombosis of the non-pathologic abdominal aorta; not only classic hypercoagulability disorders, but also immunologic, metabolic, toxicological cofactors can be involved. Delay in diagnosis is frequent and may not influence the prognosis, but does not allow conservative therapy. Prognosis depends mainly on pathologic cofactors that require detection and appropriate treatment in order to prevent complications and recurrences. PMID- 18427403 TI - Postoperative hepatic artery aneurysms development and remodeling in Ehlers Danlos syndrome type IV. Case report. AB - Patients affected by Ehlers-Danlos syndrome (EDS) type IV are at risk for aneurysm formation and rupture. This case report shows the extreme vascular fragility of these patients. We studied a 31-year-old man that developed hepatic artery aneurysms 3 weeks after splenectomy. Computed tomography angiography showed the extreme vascular remodeling of the aneurysms. We conclude that remote site complications should be kept in mind by all surgeons in vascular EDS patients even after general surgery operations. PMID- 18427404 TI - Report of a serious reportable communicable disease at a major sporting event. AB - A European Youth Olympic Sports Festival (EYOF), in Jaca, Northern Aragon, Spain, involving 1500 athletes, from 43 countries was held in January for 7 days. The event was marred by a case of type B Neisseria meningitidis. The usual care of the patient was performed in a local hospital. He eventually made a full recovery. The case was reported to the Local Health Authority and the National Public Health authority of the patient's country. Their advice was to treat the core case, give prophylactic therapy to the inner circle (28 athletes and officials) and surveillance of the other close contacts (84 total) with temperature and symptom review daily. They advised against further information being given to the rest of the athletes and officials, in case it might give rise to a panic situation. The dilemma of the responsible physicians was that 1500 athletes were traveling back to 43 countries within the incubation period of the bacterial meningitis. It was decided that informing each country was appropriate. This was done at the event by informing the Chef of the Missions and writing to the Secretary Generals of each National Olympic Committee attending. This was the first serious contagious disease at a major sporting event. The way in which the wider dispersing attendees were informed at the event should form the basis of management at sporting events in the likelihood of a serious communicable disease. PMID- 18427405 TI - Influence of work rate on dynamics of O2 uptake under hypoxic conditions in humans. AB - AIM: It was the purpose of the investigation to determine whether an altered work rate could influence the oxygen uptake (V.O(2)) and heart rate (HR) dynamics at hypoxia and normoxia. METHODS: Ten males performed a cycle exercise with 2 repetitions of 6 min each at a constant work load while breathing one of two inspiratory O(2) fractions (FIO(2)): 0.12 (moderate hypoxia) and 0.21 (normoxia). Each test began with unloaded pedaling. This was followed by three constant loads, which were 40%, 60%, and 80% of the subject's gas exchange threshold (GET) in hypoxia (F(I)O(2) = 0.12), with the 80% GET load repeated under normoxia (room air). V.O(2) was measured on a breath-by-breath basis and beat-by-beat HR via ECG, and the half time (t1/2) of each parameter was established, following interpolation data. RESULTS: There were no remarkable differences in t1/2 V.O(2) dynamics among the 40%, 60% and 80% GET; however, the differences became significant at hypoxia compared with normoxia. The HR dynamics were significantly faster in normoxia compared with hypoxia, independent of work rates. During steady-state exercise, the alterations in HR and cardiac output (Q) using the acetylene rebreathing method depended on increases in the work rate, and a significantly increase in at 80% GET was observed when compared with normoxia. Increases of stroke volume (SV) were unaffected by altered work rates and inspired O(2) concentrations. The arteriovenous oxygen difference (Ca-vO(2)) at a steady-state of exercise increased proportionally with the work rate under hypoxia, and a much greater Ca-vO(2) was observed during normoxic exercise than under hypoxia. CONCLUSION: These results seem to suggest that in humans, O(2) uptake dynamics are affected by lower O(2), not by changing work rates at hypoxia, to which the interaction between lower O(2) utilization in exercising muscles and hypoxic-induced greater blood flow can be attributed. PMID- 18427406 TI - The relative contributions of anaerobic and aerobic energy supply during track 100-, 400- and 800-m performance. AB - AIM: The present study set out to identify the relative contribution of the laboratory determined physiological measures, (maximal) accumulated oxygen deficit (AOD) and maximal oxygen uptake (VO(2max)), when predicting track performance. METHODS: Fourteen volunteers (men: n=10; women: n=4); mean (+/- standard deviation [SD]) height 1.76+/-0.1 (men) vs 1.62+/-0.08 m (women); body mass: 67.9+/-7.1 (men) vs 50.6+/-8.2 kg (women), ran track races at distances of 100, 400 and 800 m. The individually determined (maximal) AOD and VO(2max) were measured under controlled laboratory conditions (68.3+/-10.2 vs 60.7+/-16.1; men vs women, mL x (2) x Eq x kg(-1)) and (68.7+/-7.3 vs 55.6+/-4.3; men vs women, mL x kg(-1) x min(-1)), respectively. RESULTS: Track performance could be predicted using both laboratory measures, AOD and , with a high degree of accuracy: R2=76.9%, 84.8% and 89.1% for 100, 400 and 800 m, respectively. Data analysis confirmed the dominant energy supply during 100-m sprinting was the anaerobic energy supply processes, reflected as AOD. In contrast, oxidative metabolism (reflected as VO(2max)) was the dominant source of energy supply during 800-m performance. CONCLUSION: The results support earlier research, rather than present textbook dogma, namely that aerobic and anaerobic processes contribute equally to maximal exercise lasting approximately 60 s. PMID- 18427407 TI - Energy expenditure of submaximal running does not increase after cycle-run transition. AB - AIM: This study was performed to determine the relationship between increased fat oxidation and decreased running efficiency following intense cycling exercise. METHODS: Twenty-two middle-level triathletes were studied during submaximal running before and after submaximal cycling exercise. All subjects completed a 13 min run on a track at a velocity corresponding to 75% of their maximal aerobic speed (MAS) before (T1) and after (T2) submaximal cycling exercise at 75 % of maximal aerobic power (MAP). The energy cost of running (Cr) was quantified using the O(2) uptake (.VO(2)) and energy expenditure (EE) using the respiratory exchange ratio (RER). Gas exchange was measured over 30 s during the 3(rd) min and last 30 s of each run. RESULTS: The results show that after cardiorespiratory equilibration (12 min 30 s), Cr (calculated in mL(O(2))*kg(-1)*m(-1)) during T2 was higher than during T1 (+ 8.2+/-4.3%; P = 0.03). Similar observations were made for .VO(2) (+ 8.2+/-4.3%; P = 0.03) and pulmonary ventilation (+ 7.0+/ 12.3%; P = 0.04). RER decreased between T1 and T2 (- 8.6+/-9.2 %; p = 0.01). EE and Cr expressed in kJ.kg(-1).m(-1) did not vary significantly between T1 and T2. CONCLUSION: We suggest that the decrease in RER drop may be a result of greater lipid oxidation as metabolic substrate after cycling exercise. PMID- 18427408 TI - Effects of a hot environment on simulated cycling and running performance in triathletes. AB - AIM: This study examined the effects of a hot environment on metabolic responses, thermoregulation, and performance during simulated cycling and running in triathletes. METHODS: Seven male triathletes completed two trials in moderate (22+/-0.2 degrees C, 76+/-2% relative humidity, M) and hot (31.2+/-0.2 degrees C, 76.4+/-1.6% relative humidity, H) environmental conditions separated by at least 7 days. The subjects were required to complete a self-paced 40 km simulated cycling, followed by a 10 km run on a treadmill for as fast as possible in both trials. RESULTS: The overall performance time was faster in M than H (M vs H, 119+/-6 min vs 127+/-6 min, P<0.01). Moreover, there were no differences in the cycling time between the two trials, but the run time was faster in M (M vs H, 51+/-4 min vs 59+/-5 min, P<0.05) than in H. Ad libitum water consumption was higher in H than in M (H vs M, 970+/-231 mL min(-1) vs 547+/-131 mL min(-1) P<0.05), and the mean skin temperature was also higher in H than in M throughout the exercise (H vs M, 35.3+/-0.1 vs 33.3+/-0.1 degrees C, P<0.05). However, there were no differences in rectal temperature, blood lactate, blood glucose, body mass change, plasma volume change, osmolality, carbohydrate oxidation, and fat oxidation between the trials. CONCLUSION: The results suggested that triathletes reduced their running performance after a 40 km simulated cycling when the ambient temperature was high. PMID- 18427409 TI - Effects of a new experimental training program on V.O2max and running performance. AB - AIM: By testing the V.O(2max) and running performance in soccer players, the purpose of the study was to determine whether an experimental conditioning program would elicit better effects than the widely used traditional program, especially across longer distances. METHODS: Laboratory measurements of maximal oxygen consumption and maximal heart rate were performed on 48 soccer players (U19) from two first league soccer teams (Experimental team, n=24; Control team n=24). Exercise intensities were calculated from heart rate maximum. Detailed training programs for both the experimental and control groups were designed for the 13-week period. The main intervention was performed in the conditioning phase where the control group performed traditional conditioning (straight-line running, with stretching exercises in the break) and the experimental group underwent a conditioning training program on designed polygon, very much akin to a real game situation; sprint-runs with or without the ball and rapid changes in direction. During the 20, 40 or 60m sprint-run the players had to perform specific ball drills depending on marked polygon station. Running performance at 60, 100, 200, 400, 800, 1200, 2 400 m distances and V.O(2max) were tested in the initial, transitory (week 6) and final trials. RESULTS: Significant improvement in 200, 400, 800, 1200, 2 400 m running performance and V.O(2max) (P<0.01), occurred in the experimental group after only 6 weeks of program and increased more after 13 weeks. Some non-significant improvement was observed in the control group at the time of the transitory trial. However, in this group the gained improvements quickly disappeared after the beginning of the competitive season as recorded at the final trial. Discriminant analysis revealed that the groups differed the most in the 200 m variable. CONCLUSION: The new experimental program is better suited for the enhancement of V.O(2max) and running performance in soccer players and the gained improvement lasted well into the competition season. PMID- 18427410 TI - Physical load imposed on soccer players during small-sided training games. AB - AIM: The aim of this study was to examine the kinematical, physiological and technical load imposed on soccer players during three typical small-sided 3-a side training games carried out in an artificial grass 33x20 m surface. METHODS: Kinematic analysis was carried out with a bidimensional photogrammetric video system. Heart rate was recorded at 5 s intervals using Polar Accurex Plus heart rate monitors and expressed in relation to individual maximal heart rate (HR(max)). Technical parameters were registered using a specifically designed notation sheet. RESULTS: The overall intensity of these three small-sided games was superior than the experienced during competitive matches. Distance covered (747-749 vs 638 m) and mean heart rate (173 vs 166 b x min(-1)) were higher (P<0.05) in the drills without goalkeepers. The inclusion of goalkeepers reduced the tempo of the game as players performed less (P<0.05) high-intensity running and increased (P<0.05) low-intensity activities. Time spent exercising at intensities between 76-85% HR(max) was lower (P<0.05) in the game without goals, whereas the presence of goalkeepers reduced (P<0.05) activities performed exceeding 86% HR(max). CONCLUSION: The results of this study suggest that small sided games can be used effectively to develop the specific endurance capacity of football players. The integration of these drills by coaches during the regular training schedule can help replicating the demands experienced during real match play. PMID- 18427411 TI - Comparison between vertical jumps of high performance athletes on the Brazilian men's beach volleyball team. AB - AIM: The aim of this paper was to compare the anthropometric profile and the vertical jumps of two groups of Brazilian male high performance beach volleyball players. METHODS: The sample consisted of 38 male beach volleyball players from the Brazilian Beach Volleyball Circuit of 2006, allocated to two groups according to national ranking of their teams. Anthropometric measures and performance in vertical jumps were assessed using a specific methodology. RESULTS: The anthropometric results of the groups showed no statistically significant differences. The players of group 1 (G1) were better in the spike jump (P<0.01), block jump (P<0.01) and block difference (P<0.01) than the players of group 2 (G2). The prediction model of the spike jump for G2 included body mass and standing spike reach (adjusted R2=0.77) while for the block jump model it was body mass and standing block reach (adjusted R2=0.73). The regression models for G1 were not statistically significant. CONCLUSION: It is likely that vertical jump height (spike and block) influences the performance of beach volleyball players, and consequently the performance of their teams, since the present study found higher values in G1 than in G2 for the spike jump, block jump and block difference. However, an athlete's success is not related only to the variables investigated in this study; technical skill, tactics, psychology and physical conditioning can also play a role. PMID- 18427412 TI - Assessment of the reliability of a custom built Nordic Ski Ergometer for cross country skiing power test. AB - AIM: Despite the recent development of cross-country ski technique having led to an increase in the importance of upper body power, there is a substantial lack of specific equipment for upper body force and power evaluation. In order to achieve good reproducibility of the skiing motion, a new upper body ergometer has been developed in our lab and tested by elite cross-country skiers. In this study, the reliability of this device was assessed comparing upper body power measurements with double poling ski performance in the field. METHODS: The new apparatus consists of an electric motor acting as load and actively controlled by a personal computer on the basis of force and velocity data. Nine cross-country skiers (age: 21.7+/-3.12 years; body weight: 72.2+/-3.8 kg), competing at international level, performed a ski test on a 1.2 km long sprint track and a 50 s exercise on the Nordic Ski Ergometer. The velocity of the last section (180 m, slope 1.37%) of the track, performed using the double poling technique at maximal voluntary intensity, was related to the upper body power measured at the ergometer. RESULTS: Mean upper body power was 9.22+/-2.29 W kg(-1), while average velocity on the section considered was 6.66+/-0.67 m s(-1). A high correlation (R(2)=0.871) was found between upper body power and ski velocity. CONCLUSION: In addition to overcoming the main limitations that affect traditionally used equipment, the strong relationship between the parameters obtained with the new ergometer and ski velocity indicates their ability to assess athletes performance. The new apparatus could therefore be considered a ski specific testing equipment for cross-country skiers which is useful for reproducing upper body involvement in cross-country ski-ing in a laboratory setting. PMID- 18427413 TI - Cardiac and metabolic demands of in place shallow water running in trained and untrained men. AB - AIM: The aim of this study was to assess maximal cardiac and metabolic demand imposed by in place shallow water running (SWR) and to compare such responses with those obtained during treadmill running (TMR). METHODS: Twelve males, 6 trained (T) pentathletes (age: 19+/-1 years) and 6 untrained (UT) controls (age: 22+/-1 years), performed one maximal TMR test and one SWR maximal test. Heart rate (HR) and oxygen consumption (VO(2)) were continuously measured in both test conditions. RESULTS: In both groups, resting HR decreased immediately after water immersion (UT group: from 81.8+/-6.2 b x min(-1) to 69.2+/-6 b min(-1); T group: from 71.7+/-2.1 b x min(-1) to 61.7+/-7.7 b x min(-1)). In UT subjects SWR metabolic demand was comparable to that obtained during treadmill (VO(2tmr)=47.9 ml min(-1)+/-3.6; VO(2tmr)=45.2 ml x min(-1) +/- 6.8). In contrast, during shallow SWR, T subjects obtained lower V.O(2max) values with respect to TMR (V.O(2tmr)=68.9 mL x min(-1) +/-5.1; VO(2TMR)=57.2 mL x min(-1)+/-3.9; P<0.05). In the T group maximal HR was lower during SWR (from 191+/-8.1 to 177+/-7.1). At a given stride frequency, SWR was less demanding than treadmill in terms of V.O(2) and HR responses. During SWR the metabolic adjustments were depressed more than the cardiac adjustments. CONCLUSION: These findings suggest that in place SWR may be a useful exercise to reduce the effects of detraining. PMID- 18427414 TI - Muscle mass and strength, body composition and dietary intake in master strength athletes vs untrained men of different ages. AB - AIM: The aim of this study was to compare muscle strength and thickness, body composition and dietary intake between master strength athletes and controls. METHODS: Cross-sectional comparison between: 1) young control men (25.7+/-3.4 y; n=10); 2) middle-aged master athletes (52.1+/-4.7 y; n=9); 3) middle-aged control men (51.9+/-3.1 y; n=11); 4) older master athletes (71.8+/-3.8 y; n=8); and 5) older control men (70.6+/-3.3 y; n=10). Athletes had been strength trained for 22.8+/-14.9 y. Maximal isometric strength of the leg extensors was measured with a leg dynamometer, body composition by skin folds, muscle thickness of the vastus lateralis with an ultrasound scanner and dietary intake by food diaries for 4 days. RESULTS: Athletes had more lean body mass than age-matched controls (P<0.001-0.05) and young controls more than older controls (P<0.01). No group differences were observed in the thickness of vastus lateralis. Athletes showed higher absolute strength and strength per vastus lateralis thickness ratio than all control groups (P<0.01-0.001). Body mass adjusted dietary intake did not differ between the strength trained and control men. Dietary intake did not correlate with strength, muscle thickness and lean body mass. CONCLUSION: The present older men with a long-term history of strength training showed greater muscle strength and strength per muscle thickness ratio than the untrained controls. The data support the usefulness of continuous strength training to preserve muscle strength in older men. PMID- 18427415 TI - Relation between body mass index and serum aminotransferases concentrations in professional athletes. AB - AIM: Reference intervals commonly used for evaluating and interpreting laboratory values obtained in athletes are the same used in the general population. Aminotransferases (aspartate aminotransferase [AST], and alanine aminotransferase [ALT]) are commonly analyzed in serum for evaluating hepatic function. Some studies in the general population and in blood donors testified that ALT concentrations clearly correlated with weight and body mass. METHODS: We compared the aminotransferase concentrations at rest of 116 male professional athletes of 7 different sport disciplines with their body mass index (BMI). The blood drawing was performed before the start of training and of the competitive season. The athletes engaged in rugby, triathlon, soccer, sailing, cycling, basketball, alpine skiing. One hundred age-matched, apparently healthy, not physically active, males chosen for general check-up were recruited as a control group. RESULTS: The average concentrations of AST and ALT in the whole group of athletes were 24.4 U/L (standard deviation [SD]: 10.5) and 23.6 (SD: 6.5). The results in athletes were not statistically different from those of sedentary people. A positive correlation between BMI and ALT exists, whilst a very weak negative correlation between BMI and AST occurs. CONCLUSION: High concentrations of ALT should be evaluated considering BMI values whilst high values of AST should be evaluated considering the influence of physical exercise. PMID- 18427416 TI - Relationship of obesity with physical activity, aerobic fitness and muscle strength in Flemish adults. AB - AIM: The aim of this study was to analyse differences in physical activity, cardiorespiratory fitness (CRF) and muscle strength between normal weight, overweight and obese adults and to investigate the role of physical activity variables in the analyses of differences in CRF and muscle strength between these groups. METHODS: A total of 807 men and 633 women (age: 18-75 years) were included in this cross-sectional study. Weight, height, waist circumference (WC) and bioelectrical impedance were measured. Different dimensions of physical activity were assessed using a validated questionnaire. CRF (VO(2peak)) was evaluated by a maximal test on a cycle ergometer. Knee strength was measured with a calibrated Biodex System Pro 3 dynamometer. Three methods were used for classification in obesity groups: body mass index (BMI), WC and combined BMI-WC classification. RESULTS: Health-related sports and physical activity level are negatively associated with obesity in men, but not in women. Television viewing is positively associated with obesity, while VO(2peak)/fat free mass (FFM) and knee strength/FFM show a negative association with obesity in both genders. Overall, subjects with normal WC seem to be more physically active and to have somewhat better values for CRF compared to those with high WC within the same BMI category. Lower values for relative CRF and knee strength in obese subjects compared to their lean counterparts remain after adjustment for physical activity. CONCLUSION: This study confirms the lower level of physical activity and the impaired CRF and knee strength in obese adults compared to their lean counterparts. This study also sustains the importance of measuring WC and CRF during clinical examinations. PMID- 18427417 TI - BMI, fat percentage and VO2max in college female staff. AB - AIM: Low levels of physical activity and cardio respiratory fitness are both associated with higher risk of all-cause and disease-specific mortality. The purpose of this study is to examine obesity and fitness of the female staff of Ardebil Azad University in the northwest of Iran. METHODS: Thirty seven staff (medium age: 32.97+/-5.81 year, height: 158.21+/-5.88 cm, Body Mass Index [BMI]: 26.59+/-4.02 kg/m(2)) of Ardebil Azad University participated in this study voluntarily. Primary measurements of interest in the present study were height, BMI, subcutaneous skin folds, and cardio respiratory fitness determined by 1 609 meter (one mile) walk test. RESULTS: The subjects of the present study are more obese than some other population (fat percentage: 28.68+/-5.33) and cardio respiratory fitness of them is rather low (VO(2max): 33.43+/-6.90 mL/kg/min). CONCLUSION: Social/lifestyle factors such as the level of education, marital status, exercise, dietary and smoking habits may be related to overweight/obesity and cardio respiratory fitness in female staff of Ardebil Azad University. PMID- 18427418 TI - Effect of vitamin C supplementation on lipid peroxidation, muscle damage and inflammation after 30-min exercise at 75% VO2max. AB - AIM: Hypothetically, supplementation with the antioxidant vitamins C could alleviate exercise-induced lipid peroxidation. The purpose of this study was to evaluate the effect of vitamin C supplementation on exercise-induced lipid peroxidation, muscle damage and inflammation. METHODS: Sixteen healthy untrained male volunteers participated in a 30-min exercise at 75% Vo2max. Subjects were randomly assigned to one of two groups: 1) placebo and 2) vitamin C (VC: 1 000 mg vitamin C). Blood samples were obtained prior to supplementation (baseline), 2 h after supplementation (immediately pre-exercise), post-exercise, 2 and 24 h after exercise. Plasma levels of VC, total antioxidant capacity (TAC), creatine kinase (CK), malondealdehyde (MDA), total leukocytes, neutrophils, lymphocytes, interleukin-6 (IL-6) and cortisol were measured. RESULTS: Plasma vitamin C concentrations increased significantly in the VC in response to supplementation and exercise (P<0.05). TAC decreased significantly in Placebo group 24 h after exercise compared to pre-exercise (P<0.05). Although MDA levels were similar between groups at baseline, it increased significantly 2 h after exercise only in the Placebo group (P<0.05). CK increased immediately and 2 h after exercise in both groups and 24 h after exercise only in placebo group compared to pre exercise (P<0.05). Markers of inflammation (total leukocyte counts, neutrophil counts and IL-6) were increased significantly in response to the exercise (P<0.05). In VC group, there was significant increase in lymphocyte counts immediately after exercise compared with pre-exercise (P<0.05). Serum cortisol concentrations significantly declined after supplementation compared with baseline (P<0.05) as well as declined 2 and 24 h after exercise compared with immediately after exercise in VC group (P<0.05). CONCLUSION: VC supplementation prevented endurance exercise-induced lipid peroxidation and muscle damage but had no effect on inflammatory markers. PMID- 18427419 TI - Subtalar joint kinematic correlations with footprint arch index in race walkers. AB - AIM: The aim of the paper was to analyze the relationship between footprint arch index and subtalar joint movement in race walkers. METHODS: Thirteen young, highly skilled race walkers volunteered to participate. We obtained dominant-foot footprints in a bipedal stance. The arch index was measured to classify arch height. We also conducted a photogrammetric video-3D study on a running track. The support phase was recorded while subjects race walked at their individual competition speed. We calculated 4 angle time series describing the ankle joint kinematics during the support phase. Five specific step instants were calculated for each angle and correlated with the arch index. Race walkers were grouped according to arch height to compare. We also correlated the arch index with the time in medial support, and time to change from lateral to medial support during the stance phase. RESULTS: In the calcaneal angle we found correlations with the footprint (r=0.81; P<0.01) and statistical differences when grouped (P<0.01). In this angle we also found a moderate-high correlation between footprint arch index and the time in medial support (r=0.77; P<0.01), and also in the time to change from lateral to medial support during the stance phase (r=-0.73; P<0.01). Data also show that the race walkers adopt a characteristic propulsion technique in the end of the support. No statistical differences were found in the rearfoot angle, which has been previously associated to specific injuries in running. CONCLUSION: In conclusion, race walkers with higher arches exhibit a more pronounced support with the lateral side of the foot and they do so for a longer time. Conversely, subjects with flatter feet support with the medial side of the foot. The footprint has been found to be a good predictor for the technique employed with respect to the medial and lateral strike of the foot reflected by the calcaneal angle. Coaches should keep this in mind from both performance and injury prevention viewpoints. PMID- 18427420 TI - Effects of percutaneous transluminal coronary angioplasty on cardiopulmonary responses during exercise. AB - AIM: The aim of this study was to examine by means of percutaneous transluminal coronary angioplasty (PTCA) whether dilating blocked coronary arteries in patients with coronary heart disease will improve relevant attributes measured during cardiopulmonary exercise testing (CPET). METHODS: Fourteen myocardial ischemia patients (13 male, 1 female; age range: 42-72 years), who were referred for cardiac catheterization, participated in the study. Eight patients underwent PTCA (experimental group) and 6 did not (control group). All subjects performed CPET about 2 weeks before and 3 weeks after cardiac catheterization. RESULTS: The results of electrocardiogram (ECG) analysis reveal a very low predictive accuracy (36%) for identifying coronary heart disease and/or anatomical changes caused by the PTCA. However, selected CPET parameters in the experimental group only showed significant post-PTCA changes from baseline. Peak oxygen uptake (VO(2)) increased from 17.49 to 20.75 mL x kg(1) x min(1), ventilatory anaerobic threshold from 12.15 to 14.39 mL x kg(1) x min(1), peak oxygen pulse (O(2)pulse) from 11.76 to 13.27 mL x beat(1), and O(2)pulse slope from 7.05 to 9.25, slope category. No significant inter- or intra-group differences were found in peak exercise heart rate, respiratory exchange ratio, subjective rating of perceived exertion, or systolic and diastolic blood pressure. CONCLUSIONS: This study suggests that selected CPET parameters seem to be highly sensitive to changes in cardiac function caused by the PTCA, significantly more than with conventional stress ECG. These findings merit further investigation. PMID- 18427421 TI - Effect of exercise training on cutaneous microcirculation in rats. AB - AIM: Exercise training is known to improve endothelium-dependent relaxation in the coronary and skeletal muscle arteries. However, the effects of exercise training on peripheral nonworking tissue, including microcirculation, are still unclear. Therefore, we investigated the effect of chronic and regular aerobic exercise on cutaneous microvascular endothelial function in rats. METHODS: We assessed the effect of physical training on skin microcirculation in 7 sedentary (SED) and 21 training rats (Wistar-Kyoto), submitted to a treadmill training protocol (15 m/min; 15% incline; 60 min/day; 8 weeks). Training rats were divided into 3 groups, exercising 1 day/week (Ex1), 3 days/week (Ex3) or 5 days/week (Ex5). Cutaneous blood flow was recorded before the beginning of the training protocol, after 4 weeks and at the end of the training program. Hyperemic response (RH) was the flow reaction obtained after sudden release of the garrot. For data analysis, cutaneous vascular conductance (CVC) was indexed as cutaneous blood flow divided by mean arterial blood pressure (in millimeters of mercury, mmHg) and normalized to baseline values. RESULTS: At baseline, CVC was not different among groups (SED or training) at 3 steps of experimental protocol. The hyperemic stimulus significantly increased normalized CVC only in group Ex3 after 4 weeks (P<0.006) and 8 weeks (P<0.006). CONCLUSION: Exercise training exerts a generalized effect on the vasculature by increasing endothelial function in vessel beds different from those perfusing actively working muscle. However, some differences exist since training at a frequency of 3 bouts weekly only modifies cutaneous microcirculation. PMID- 18427422 TI - Insulin resistance and glucose tolerance in obese women: the effects of a recreational training program. AB - AIM: This study was designed to investigate whether recreational physical activity based on aquatic training may improve lipid profile and glucose tolerance in obese subjects. Additionally, we intended to assess a possible correlation between insulin resistance index HOMA(IR) with another index including serum adiponectin concentrations. METHODS: Study population consisted of 12 obese women aged 44-61, who volunteered for an aquatic training 1 h twice a week for 3 months. Before entering the training program, and after completing it patients' oral glucose tolerance test was performed. Glucose, insulin and adiponectin concentrations, levels of antibodies to oxidatively modified low density lipoproteins (oLAB), and lipid profiles were measured in blood samples collected before the oral glucose tolerance test (OGTT). Glucose and insulin concentrations were also assessed in blood samples collected at 30 and 120 min of OGTT. Atherogenic index of plasma (AIP), and insulin sensitivity indexes HOMA(IR) and HOMA(AD) were calculated on the basis of the obtained data. RESULTS: The 3 month, recreational aquatic training had no influence on body mass, but it resulted in improved glucose tolerance (at t = 0 and t = 120 min), decreased index (HOMA(IR)), and decreased levels of total and LDL-cholesterol (P<0.05). Adiponectin concentrations and values of HOMA(AD) remained unchanged. We found a significant correlation between adiponectin levels and concentrations of total and LDL cholesterol, and between insulin indexes HOMA(IR) and HOMA(AD) both before, and after the training (P<0.01). CONCLUSION: The results of our study indicate that a 3-month recreational training consisting in water aerobic results in favorable changes in glucose and lipid metabolism in obese subjects, even despite the lack of improvement in body mass. PMID- 18427423 TI - The effects of swimming exercise on high-fat-diet-induced steatohepatitis. AB - AIM: The aim of this study was to assess the effects of swimming exercise on high fat (HF) diet-induced steatohepatitis. METHODS: Two groups of rats were fed a HF diet (35%; kcal) for 12 weeks, with one group remained sedentary (HF-Sed) and the other group swam (HF-Tr) during this entire period. A third group was fed a standard diet and remained sedentary for 12 weeks (SD-Sed). Swimming exercise (5 days/week) was initiated at the same time as the HF diet and was progressively increased, until it reached 90 min after 4 weeks. RESULTS: At the end of the 12 week period, HF-Sed rats exhibited higher body weight (means +/- standard error [SE]: 492+/-40 vs 432+/-21 g; P<0.05), liver weight (16.21+/-2.23 vs 9.92+/-1.14 g; P<0.05), serum alanine aminotransferase (73.62+/-36.11 vs 27.16+/-4.62 U/L; P<0.05), aspartic aminotransferase (AST; 198.87+/-120.16 vs 120.16+/-19.09 U/L; P<0.05), nonesterified fatty acids (NEFA; 897.48+/-133.62 vs 437.5 micromol/L; P<0.05), triglyceride (0.92+/-0.1 vs 0.56+/-0.04 mmol/L; P<0.05), tumor necrosis factor (TNF)-a (21.74+/-2.93 vs 16.57+/-3.53 pg/mL; P < 0.05) as well as liver malondialdehyde (MDA; 5.3+/-1.45 vs 3.48+/-0.94 nmol/mg protein; P<0.05) and NEFA (305.47+/-69.23 vs 204.56+/-64.26 micromol/mg protein; P<0.05) than SD-Sed rats. All HF-Sed rats developed moderate to severe hepatic steatosis and steatohepatitis according to liver histological findings. Compared with HF-Sed, HF-Tr rats' hepatopathological manifestations of steatosis and inflammation were much attenuated, and their serum and liver parameters were all lower with the exception of serum AST. CONCLUSION: It was concluded that swimming exercise might reduce the incidence of HF-diet-induced steatohepatitis. PMID- 18427424 TI - Impact of acute exercise on bone turnover and growth hormone/insulin-like growth factor axis in boys. AB - AIM: The aim of this study was to investigate the response of N-terminal propeptide of type I procollagen, crosslinked telopeptide of type I collagen and the growth hormone/insulin-like growth factor-I axis to acute aerobic exercise in boys at different pubertal stages METHODS: The subjects were 60 healthy boys (group I - Tanner stage 1, N=20; group II - Tanner stages 2 and 3, N=20; group III - Tanner stages 4 and 5, N=20) who exercised 30 minutes at constant load on cycle ergometer at the level of ~95% of their individual ventilatory threshold. Venous blood samples were obtained before, immediately after and after 30 minutes of recovery for the measurement of serum testosterone, growth hormone (GH), insulin-like-growth factor-I, insulin-like-growth factor binding protein-3, N terminal propeptide of type I procollagen (PINP) and crosslinked telopeptide of type I collagen. RESULTS: Acute exercise did not affect significantly serum testosterone, insulin-like-growth factor-I, insulin-like-growth factor binding protein-3 or bone turnover markers concentrations in any of study groups. The rise in growth hormone concentration during exercise was highest in group III (62.3+/-41.7 mU/L vs 15.5+/-11.4 in group I and 41.8+/-20.0 in group II). The increment in serum growth hormone level during exercise was positively correlated (r=0.64; P<0.001) to basal serum testosterone concentration. CONCLUSIONS: It can be concluded that growth hormone response to exercise was directly dependent on serum testosterone concentration. Acute exercise did not affect serum testosterone, insulin-like-growth factor-I, insulin-like-growth factor binding protein-3 or bone markers levels. PMID- 18427425 TI - Altered perception and report of fatigue and recovery in veteran athletes. AB - AIM: This study investigated whether ageing effects perceived and reported ratings of fatigue and total quality of recovery following high-intensity training in athletes. We hypothesized that veteran (V) athletes would report greater changes in perceived measures of fatigue and recovery than training matched younger athletes. METHODS: Perceptions of muscle soreness (SOR), fatigue, and recovery were recorded in young (Y) and V (>35 years) well-trained cyclists in response to 3 days of repeated cycling time trials. Nine Y (24+/-5 years) and 9 V (45+/-6 years) cyclists performed 3 consecutive days (T1-T3) of 30-min cycling time trials (TT30) intended to induce fatigue leading to decreased performance. Physiological and performance variables were measured before, during, and after each time trial. Subjective measures of SOR, fatigue, and recovery were recorded each day. RESULTS: There was no change in performance at the TT30 from T1 to T3 for either group. SOR, fatigue, and recovery significantly changed over the 3 days in the V group, but not in the Y group. The change in SOR from T1 to T3 was significantly greater in the V group than in the Y group (22+/ 14 mm vs 9+/-12 mm, respectively; P=0.04). CONCLUSION: It was concluded that 3 days of cycling time trials induce perceptions of muscle pain/SOR, fatigue and reduced recovery in well-trained V cyclists with no corresponding decline in physical performance. PMID- 18427426 TI - Extracranial-intracranial bypass for giant aneurysms and complex vascular lesions: a clinical series of 10 patients. AB - AIM: The purpose of this clinical study is to evaluate the preventive bypass indications, the kind of bypass to use, and the timing of postoperative internal carotid artery (ICA) occlusion in the treatment of complex aneurysms. METHODS: This study enrolled 10 patients suffering from complex aneurysms treated using several kind of extra-intracranial bypass on the basis of preoperative careful assessment of cerebrovascular flow. This assessment is based on the balloon test occlusion (BTO), from which three different conditions emerge. According to the Department of Neurosurgery these three different conditions require three different kinds of bypass: in case of depleted flow a short STA-MCA bypass is necessary, in case of intermediate flow a long arterial graft bypass is required, whereas in case of high flow a long venous graft bypass is mandatory. RESULTS: Bypass patency was found in six out of seven patients who underwent surgery for giant ICA aneurysms, as well as in other patients. One of the patients died for a septic shock, another had a neck hematoma that required surgical evacuation. At mean follow-up no further neurological deficits were found. CONCLUSION: The severe complications due to the combined approaches can be reduced thanks to the indications obtained by BTO concerning the kind of bypass to perform and to the timing of the ICA occlusion. PMID- 18427427 TI - The pillar pain in the carpal tunnel's surgery. Neurogenic inflammation? A new therapeutic approach with local anaesthetic. AB - AIM: After the surgical decompression of the transverse carpal ligament as treatment of the tunnel carpal syndrome, pillar pain manifestation is possible. This is a painful and temporary invaliding syndrome with unknown aetiology. Aim of the study is to demonstrate that pillar pain is based on autonomic irritation that disappears with simple infiltration of local anaesthetic (LA), or rather by neuromodulation. METHODS: Eighty-four patients were enrolled for this study and underwent an open release technique surgery. They were then asked for regular postoperative follow-ups. RESULTS: Thirty-two out of 84 patients (38%) developed pillar pain. Accordingly, injection of LA as pain treatment has since been studied and results compared with the conventional protocol for this painful syndrome. CONCLUSION: Even if the number of the patients considered is not great, there is evidence of a decrement of pillar pain by means of LA injections. Excellent functional outcomes and satisfaction were achieved using LA infiltrations for pillar pain after carpal tunnel decompression. The minimally invasive technique offers a quick, easy, effective, and inexpensive method useful to minimize and cure in a few days the pillar pain. PMID- 18427428 TI - Therapeutic criteria in communicating childhood hydrocephalus. AB - AIM: The aim of this study was to evaluate the usefulness of cerebral blood flow velocity in the middle cerebral artery measured by transcranial Doppler as criteria to therapeutic action in communicating hydrocephalic children. METHODS: In eight non-tumoral communicating hydrocephalic infants, ranging from five to 18 months of age, monitored from 18 to 36 months (mean time of follow-up: 24.25 months), cerebrospinal fluid (CSF) oxypurines (hypoxanthine and xanthine) and uric acid levels were compared by means of the Evans' index, the mean weekly increase in cranial circumference, and the transcranial Doppler measurements. RESULTS: Results indicate that clinical (mean weekly increase in head circumference), radiological (Evans' index), biochemical (oxypurines and uric acid in the CSF), and hemodynamic (transcranial Doppler) criteria have the same role in monitoring infantile hydrocephalus. CONCLUSION: In conclusion the transcranial Doppler measurement can be done noninvasively and examinations can be repeated when needed, obtaining immediate RESULTS: Hence, it is the most adequate monitoring technique in clinical practice. PMID- 18427429 TI - Implantation of a hybrid solution, using a combination of instrumentation systems for the treatment of degenerative disorders of the lumbosacral spine. A technical note. AB - This article describes a novel hybrid technique for implantation of a combined spinal instrumentation system incorporating lumbosacral facet-joint screw fixation and single- or two-level placement of a lumbar interspinous distractor (X STOP). This system accomplishes two main tasks: 1) immediate rigid fixation of the lumbosacral junction; 2) single- or two-level lumbar decompression with no opening of the spinal canal or intervertebral foramina. This technique may be a reasonable alternative to traditional surgery for patients suffering from multisegmental degenerative disorders of the lumbosacral spine. PMID- 18427430 TI - Pelvi-ureteric junction obstruction and renal function after pyeloplasty: a retrospective study in 29 children. AB - AIM: Actually, unilateral obstruction is not indicated for surgery, especially if renal function is unaffected. This retrospective study focused on the renal function after pyeloplasty for unilateral obstruction in children. METHODS: Twenty-nine children were retrospectively reviewed. To compare the ultrasound readings in patients with different ages, the comparative-length-index (index) of each renal unit (RU) was calculated. MAG3 dynamic scintigraphy was applied to diagnose any obstructions. RESULTS: Children with left obstruction were younger than children with right obstruction, when surgery was performed. Scintigraphic scan of right RUs with obstruction operated later was a lower at diagnosis than jet of the normal contralateral. Normal scintigraphic scan at diagnosis of left RUs with obstruction operated early revealed that RU was slightly reduced after the operation. At both diagnosis and follow-up the index between obstructed RUs and normal contralateral was comparable, even if it was significantly higher in left obstructed RUs. At follow-up, however, the scintigraphic scan revealed that RUs were more reduced in patients who were treated late. CONCLUSION: Early pyeloplasty partially safeguard kidney function in children with unilateral obstruction, whose renal function was normal at birth. The reduced kidney function observed before surgery was not proportionally improved after surgery with respect to the contralateral that was not affected. PMID- 18427431 TI - Intraoperative double-J stent insertion in children with scintigraphic impaired renal function and obstructive urinary tract malformation. AB - AIM: Ureteral double-J (DJ) stents are frequently used in modern urologic practice. At present the role of stents in urological and surgical practice and their efficacy in paediatric age are not yet clear. The aim of this study is to evaluate advantages and efficacy of ureteral stents, correlating clinical and radiological data with the permanence of stent in situ. METHODS: Between July 1999 and July 2004 surgery with ureteral stenting was performed on 24 consecutive patients aged between 2 and 13.5 months with scintigraphic impaired renal function due to an obstructive urinary tract malformation. During the study the performance and the efficacy of indwelling stent have been evaluated through clinical and radiological variables: pre-, intra-, and post stenting blood tests, ultrasonographic and scintigraphic parameters were also evaluated. RESULTS: The stent insertion was useful to improve renal parenchymal thickness and renal growth. No correlation was found between improved blood tests and scintigraphic values. The improvements of clinical and radiological data were strictly correlated with the time of stenting (>3 months). CONCLUSION: The insertion of DJ stents as long-term internal urinary diversion is useful and safe. Late complications related to the use of stents are not frequent. PMID- 18427432 TI - Manipulation of androgens and alterations in the androgen receptor axis in prostate cancer. AB - Androgens play a key role in prostate cancer (PCa) development and progression. For decades, androgen deprivation therapy (ADT) has been the primary treatment for men with metastatic prostate cancer. ADT initially results in the regression of prostate cancers, however, this effect is limited and patients inevitably develop an androgen refractory form of the disease. Many studies have implicated the androgen receptor (AR), the key mediator of androgen action, as an important player in PCa recurrence. Changes such as AR gene amplification, AR mutations, and in AR subcellular localization, and AR coregulator status that result in the upregulation of AR activity are often associated with androgen refractory prostate cancers. This review seeks to highlight some of the changes in the AR axis in androgen refractory tumors and the recent advances in ADT. A greater understanding of the AR axis will provide a basis for improving existing methods and developing new methods for the treatment of prostate cancer. PMID- 18427433 TI - Robotic assisted laparoscopic radical prostatectomy: evolution and outcomes. AB - Robotic assisted laparoscopic prostatectomy (RALP) is rapidly gaining acceptance in the urologic community as an effective treatment modality for the treatment of localized prostate cancer. For the open surgeon, this technology assists in the transition to laparoscopic-based surgery. With early stage cancer, the absence of tactile feedback appears to be offset by the superior visualization and instrumentation that the daVinci robot (Intuitive Surgical, Sunnyvale, CA, USA) provides. Economical considerations also appear to be outweighed by patient demand and benefits. Short-term perioperative, functional, and oncological outcomes all appear to be equivalent to the gold standard, open radical retropubic prostatectomy. In conclusion, RALP is an excellent option for patients considering prostate cancer treatment and should be part of the urologic surgeon's armamentarium. PMID- 18427434 TI - Management of lymph nodes in early prostate cancer. AB - This article is a review of the literature concerning the management of lymph nodes (LNs) in early prostate cancer. The literature review was performed using the PUBMED database. The treatment of LNS in early prostate cancer usually requires a pelvic LN dissection (PLND), which is performed in all patients with localized prostate cancer. Sometimes nomograms are used to select patients requiring lymphadenectomy. No consensus is available concerning the indications for the performance of PLND and the extent of the lymphadenectomy that should be performed. Recent non-surgical methods detecting LN invasion seem promising, but further investigations and a consensus attainment for terminology and investigation methodologies are advisable. PMID- 18427435 TI - Biomarkers for early prostate cancer detection. AB - In the 1990s the discovery of prostate-specific antigen (PSA) revolutionized early prostate cancer detection. Since that time, PSA has become an indispensable marker for diagnosis and follow up of prostate cancer patients. Despite its remarkable performance, PSA is not cancer specific. High PSA levels are found in both cancerous and healthy tissue, particularly in benign prostate disease, resulting in significant numbers of false positive cases. Hence, there is a need for new markers that better differentiate benign from malignant lesions and indolent from aggressive cancers to decrease the potential over treatment of prostate cancer. With recent advances in biotechnology, many promising blood biomarkers have been identified and are currently under investigation. This article reviewed the literature searching for emerging biomarkers for early prostate cancer detection. PMID- 18427436 TI - [Cardiovascular and urological combined operation in a patient with severe aortic stenosis and renal cell carcinoma]. AB - The simultaneous treatment of cardiovascular and urological diseases is at present discussed concerning indication and the increasing postoperative complications. Rarely these cases are observed with simultaneous critical diseases for both apparatuses thus conditioning the manner and timing of procedures. Authors report a case of sequential cardiosurgical and urological treatment in a patient affected by severe aortic stenosis and kidney neoplasm with bleeding. Authors performed the combined approach in the cardiosurgical department, starting with the aortic valve replacement with extracorporeal circulation (ECC) procedure and subsequently with the radical right nephrectomy. We believe that, with limitations according to each case, this approach can be taken into account in selected cases with severe cardiac valve disease refractory to medical therapy with persistent hematuria or bleeding. PMID- 18427437 TI - Lymphangioma communicating with the excretory system. AB - Lymphangioma is a rare benign tumor caused by failure in the development of the lymphatic communicating system. The corresponding nomenclature is confusing. In recent years ''renal lymphangiectasia'' is the preferred name. Although this disease may occur in any site of the body, the neck (75%) and axillary area (20%) are the most common sites, and the kidney is occasionally involved. We report a case of lymphangioma communicating with the urinary system in a 61-year-old man diagnosed by CT scan treated with nephrectomy and histological confirmation. PMID- 18427438 TI - Postchemotherapy resection of a metastatic retroperitoneal nonseminomatous germ cell tumor of the testes. PMID- 18427439 TI - Male breast cancer: 6-year experience. AB - AIM: Breast cancer in men is a very rare neoplasm accounting 1% of all breast cancer with an incidence ratio of 1:100 of men to women and about 1% of all malignancies in men. On the basis of the literature review the authors tried to determine the main characteristics of this rare neoplasm in terms of epidemiology, diagnosis, prognosis, treatment and survival. METHODS: The authors report the experience of the Breast Unit of the San Giovanni Addolorata Hospital in Rome, where 4 cases of male breast cancer were observed and treated over 784 breast cancers. RESULTS: All tumours were ductal carcinomas. The extent of disease was as follows: 3 cases with stage I and 1 case with Stage IIIB; in two cases estrogen and progesterone receptors expression was 100% and in the other two cases it was 20-80%. Median follow up was 57.5 months. At present, after 6 year follow up the three patients with stage I are in good conditions; the patient with stage III died after 27 months with metastatic disease. CONCLUSIONS: Surgical treatment remains the gold standard in male breast cancer. The prognosis for males with breast cancer is similar to female patients on equal terms of stage of disease. Adjuvant therapy is based on retrospective studies of male breast cancer conducted over the past 20 years using the guidelines for breast cancer in women. PMID- 18427440 TI - Maxillary sinus lift through heterologous bone grafts and simultaneous acid etched implants placement. Five year follow-up. AB - AIM: The aim of this study was to assess retrospectively at 5 years the clinical outcome of bone regeneration in patients who underwent maxillary sinus lift with different equine-derived, enzyme-deantigenated equine bone grafts, and simultaneous placement of acid-etched surface implants. METHODS: Eighteen patients (10 males and 8 females, age between 55 and 61 years) were considered. They were divided into 2 groups according to the type of bone graft used: a combination of an equine flexible heterologous spongy bone layer (Osteoplant Flex, Bioteck) and equine heterologous cortical and spongy granules (Bio-Gen Mix, Bioteck), group A, or a block of equine heterologous spongy bone (Bio-Gen Block, Bioteck), group B. Surface-treated implants (''TRE'' type, Biotec) were placed simultaneously to sinus augmentations and usual clinical and radiological tests were performed at 6 months and every year up to 5 years after surgery. RESULTS: At 5 years, 45 out of 49 total implants placed (91.8%) were successful according to the Albrektsson and Zarb criteria. The loss of the 4 failed implants was observed in 2 patients of group B during the first days following surgery. CONCLUSION: These results show that the heterologous bone grafts supported properly the bone regeneration inside the sinuses, and that the type of implants used allowed to achieve a satisfying rate of success according to the current standards in implantology. PMID- 18427441 TI - [Gastric cancer: surgical treatment and prognostic score]. AB - AIM: Gastric cancer is the fifth most common cause of tumor-related death in Western countries. Surgery is the only effective treatment but only 50-60% of patients can receive a curative treatment because of absent or aspecific symptoms. The aim of this study was to develop a scale for gastric cancer patients that takes into account factors related to the tumor and to the patient. METHODS: Fifty-seven patients with gastric adenocarcinoma admitted to the Department of General, Geriatric Surgery and Diagnostic and Operative Endoscopy of the University ''Federico II'' in Naples, and treated by gastrectomy from January 1998 until December 2002, were included in this retrospective cohort. The prognostic score was created according to the variables identified in Cox analysis as statistically significant (P 0.1). RESULTS: The 5-year mortality rate was 61%. Cox analysis identified these variables with a significant effect on mortality: age ?60 (odds ratio (OR) 4.16; P=0.015), smoking or alcoholism (OR 2.66; P=0.057), pTNM I (OR 0.04; P=0.003), pTNM II (OR 0.18; P=0.029), pTNM III (OR 0.27; P=0.023), pTNM IV (OR 3.28; P=0.012), lymph node ratio (LNR) <20% (OR 0.15; P=0.01), LNR 20% (OR 3.83; P=0.002), Lauren diffuse histotype (OR 2.41; P=0.1) and location of the neoplasm at superior third (OR 6.70; P=0.003), middle third (OR 5.60; P=0.003), or inferior third (OR 0.32; P=0.008). Patients have been randomized into three groups according to their scores (3-40.5; 41-78.5; 79 115.5) and the 5-year mortality rate was 46%, 59%, 90% in group 1, 2 and 3 respectively. CONCLUSION: It is necessary to consider in prognostic stratification of gastric cancer patients not only pTNM staging but also other factors such as age, smoking or alcoholism, Lauren histotype, location and linfonodal involvement. It is possible to design a more effective prognostic score predicting the individual risk and addressing the therapy and the follow up. PMID- 18427442 TI - Characteristics and prognosis of resected T3 non-small cell lung cancer. AB - AIM: The aim this study is to determine the characteristics, survival, and factors affecting the survival of resected T3 non-small cell lung cancer. METHODS: The records of 97 cases were retrospectively reviewed with T3 non-small cell lung cancer patients that were operated between 1996-2001. Complete resection was achieved in 71 (73.2%) patient. The distribution of N status was 47.4%, 28.9%, and 23.7% for N0, N1 and N2 respectively. The evaluated prognostic factors in univariate and multivariate analyses were, histologic type, type of resection, N status, subgroups of pT3, resection margins and effect of adjuvant therapy. RESULTS: Overall 5-year survival rate was 24.3%. Median survival and 5 year survival of the patients whose tumors resected completely was 33 months and 31.5%, whereas 18 months and 7.3% for the patients resected incompletely (P=0.03). Median survival being not significantly different among the three subgroups: 25, 23, and 32 months (P=0.7) in the bronchial pT3, mediastinal pT3, and peripheral pT3 subgroups, respectively. Histology (P=0.57), type of surgical resection (lobectomy versus pneumonectomy) (P=0.25), and use of adjuvant therapy (P=0.054) did not influence the survival significantly. However N status influenced the survival significantly (P=0.01). According to the multivariate analyses, two factors were selected as prognostic indicators: N2 status (P=0.03) and incomplete resection (P=0.03). CONCLUSION: Three pT3 subgroups did not show survival differences. Complete resection and N2 status are the two most influencing factors in survival of the patients. Adjuvant therapy effected the survival and the quality of life reversely. PMID- 18427443 TI - Acute appendicitis: a descending trend? AB - AIM: The diagnosis-related group (DRG) system is a prospective hospital payment system used to categorize hospital patients expected to require similar hospital services. In Italy, hospital productivity is calculated from DRG-based data coded according to the International Classification of Diseases, Ninth Revision, Clinical Modification (ICD-9-CM), which is a classification system for coding of diagnoses and operations for indexing medical records by diagnosis and operations. The aim of our survey was to determine the national incidence of appendectomies based on the coded summary of selected data in hospital discharge reports (HDR). METHODS: The Italian Ministry of Health online database was searched for data collected between 2001 and 2003. The search engine allows analysis by different regions but not by individual hospital. The national incidence of appendectomy was calculated using data collected from the ICD-9-CM and from the HDR. In a deeper analysis, regional data and data from individual hospitals were compared. RESULTS: The analysis revealed the incidence of appendectomy, rates of simple acute appendicitis vs complicated appendicitis, common laparotomic appendectomy vs laparoscopic appendectomy, as well as mean duration of hospitalization. CONCLUSION: The incidence of acute appendicitis has considerably decreased, whereas the rates of complicated appendicitis have increased because of longer diagnostic and therapeutic delay, inappropriate antibiotic therapy and upclassifying of diagnosis and procedures in the HDR (ICD 9-CM) in order to obtain a ''wider impact'' on DRG. PMID- 18427444 TI - Minimally invasive techniques in colon surgery. AB - With the description of the first laparoscopic cholecystectomy in 1985, minimally invasive approaches have become the standard practice of surgeons in managing several disease processes. This has been mainly driven by the significant favorable impact of minimally invasive surgery on patient related outcomes. Smaller incisions lead to improved cosmesis, reduced postoperative pain and earlier return of gastrointestinal function. These factors in turn contribute to a faster recovery of the patient (compared to similar open procedures) with a reduced utilization of hospital resources, reduced costs and earlier return of the patient to normal routines of daily life and work. With experience it is clear that these favorable patient outcomes can also be seen with minimally invasive surgery for various colonic diseases and procedures. Many of the early concerns regarding minimally invasive approaches such as port site recurrence and the feasibility of adequate oncologic resections have been laid to rest by multiple randomized trials. There are now documented benefits to minimally invasive approaches for colonic diseases such as cancer, inflammatory bowel disease and diverticular disease; as long as surgeons choose the appropriate patients and spend the time and resources needed to become proficient at these advanced procedures. PMID- 18427446 TI - Laparoscopic surgery for inflammatory bowel diseases. AB - Since the introduction of laparoscopy into colorectal surgery in the early 1990s, almost every procedure was attempted laparoscopically. Performing laparoscopic colectomies in patients with inflammatory bowel diseases (IBD) might prove to be extremely challenging due to the inflammatory features of the diseases and the potential complications they may pose; dealing with inflammatory masses, fistulas and abscesses, short and thick mesentery, friable bowel wall, and the need to operate in all the abdominal quadrants, mobilizing long segments of large and small bowel and controlling multiple large blood vessels is not an easy task. Consequently, many very experienced surgical groups conducted numerous trials in an attempt to determine whether laparoscopy in IBD is indeed beneficial or not. The focus of this review is minimally invasive procedures in patients with ulcerative colitis (UC) and Crohn's disease (CD). PMID- 18427445 TI - Minimally invasive approach to colorectal surgery. AB - While the development of laparoscopic surgery over the last two decades was amazingly fast, its adoption was neither uniform nor universal. Some procedures, like laparoscopic cholecystectomy, rapidly became the standard of care throughout the surgical community. Laparoscopy for colorectal surgery gained much less acceptance. Factors such as technical complexity, cost, duration of surgery and concerns about oncologic safety influenced the hesitancy in performing this surgery, and it took the surgical community more than a decade to admit that the laparoscopic option is legitimate: it is safe, and it provides the patients with the advantages of minimally invasive surgery, without any surgical or oncologic compromise. This slow process of maturation had a significant advantage, as it allowed this kind of surgery to be thoroughly investigated. Its acceptance is now well based on multitude of data, available from many basic science and clinical studies. Not many procedures in the daily surgical practice are as evidence-based as is laparoscopic colon surgery. The aim of this review was to describe some general aspects of laparoscopic colorectal surgery, and examine the data supporting its use in different procedures for various pathologies, both benign and malignant. PMID- 18427447 TI - Laparoscopic reintervention in colorectal surgery. AB - Laparoscopic colorectal surgery has developed in the 1990's and beginning of 2000. The favourable results and great progress in the development of laparoscopic techniques have expanded the indications of laparoscopic colorectal surgery. More and more complicated colorectal cases are treated laparoscopically, including those having had previous laparotomies. Surgical reinterventions after colorectal procedures are common. Reinterventions are either intended to treat complications of colorectal surgery or to treat colorectal disease after previous abdominal or pelvic surgery. Laparoscopic reinterventions face surgeons with specific challenges related to morphological changes in the abdomen. Adhesions are primarily responsible for these changes and evoke various complications such as trocar injury, bleeding, enterotomy and conversion to laparotomy. Trocars and Veress needle are responsible for up to half of all bowel injuries in laparoscopic surgery and adhesion formation is the most important risk factor for bowel injury. The risks of adhesions are often underestimated. The first clinical results on laparoscopic reinterventions are promising. Routine use of anti adhesion agents and diagnostics is advocated to prevent adhesion formation and make reintervention more safe reducing serious complications as inadvertent enterotomy, bleeding and trocar injuries. More research is needed to develop better tools for mapping adhesions, as none of the trocar placing techniques can rule out bowel injury. Improved diagnostic tools for mapping adhesions will also facilitate patient selection for laparoscopic treatment of SBO. PMID- 18427448 TI - [A rare presentation of Amyand's hernia. Case report and review of the literature]. AB - Amyand's hernia is defined as an inguinal hernia within the hernial sac containing the appendix. It is a rare disease, reported in 1% of cases of inguinal hernia repair. The appendix can be complicated by acute appendicitis in 0.13% of cases. This disease is often very difficult to diagnose, and most of the time it can be confused with an incarcerated or strangulated inguinal hernia. Often, it requires an emergent surgical treatment. This article describes the case of a 82-year-old female who was admitted for an intestinal obstruction and a bulge in the right inguinal region. An abdominal computed tomography scan showed dilated small bowel loops with multiple air/liquid levels and one loop herniating into the right inguinal canal. The patient underwent a laparotomy that showed the presence of an acute appendicitis and a necrotized ileal loop protruding into the right inguinal canal. The patient underwent an appendectomy and small bowel resection and she was discharged on postoperative day 10. Amyand's hernia can be a challenge for the surgeon. Its treatment depends on the grade of inflammation of the appendix. In fact, it can range from the simple repair of the abdominal defect with a prosthetic mesh, to appendectomy, small bowel resection and repair of the abdominal wall defect without a mesh. PMID- 18427449 TI - Sequential left pneumonectomy and right upper lobectomy for hemoptysis in post tuberculosis destroyed lung and aspergilloma. AB - The case of a 30-years-old man from Angola who was referred with a history of previous tuberculosis at the age of 14, recurrent hemoptysis and a radiological picture of destroyed left lung and shrunken right upper lobe containing a fungus ball is presented. After careful functional evaluation the patient underwent sequential left pneumonectomy and right upper lobectomy due to the fear of massive and possibly fatal hemoptysis. Both operation were well tolerated and had an uneventful postoperative course. There was no significant difference between his postoperative and his preoperative functional status. The patient is now living an active life with only seven lung segments. PMID- 18427450 TI - Unusual paravertebral location of bronchogenic cyst. PMID- 18427451 TI - Uterine artery pseudo-aneurysm during pregnancy: diagnosis and surgical treatment. PMID- 18427452 TI - [Hemorheology and homeostasis in the most acute stage of lacunar stroke]. AB - Eighty patients were studied at the first day of the development of lacunar stroke, the main attention being focused on the particular form of focal pathology--a small deep (lacunar) stroke (SDS). Patients with arterial hypertension and complication free course had higher hemorheologic parameters (blood viscosity, erythrocytes aggregation) and a shift of prostacyclin thromboxane balance towards preaggregation and decrease of fibrinolysis. These disturbances strengthened during the development of SDS. The endothelium dysfunction, alterations of blood rheologic characteristics due to the formation of aggregants of erythrocytes, decrease of erythrocytes deformation and activation of cell homeostasis play a central role in the development of intracranial arterial occlusion resulted in the SDS development. At the same time, fibrinolysis remains relatively stable. PMID- 18427453 TI - [The problem of comorbidity of phobic disorders in adolescents]. AB - Age peculiarities of the comorbidity of the phobic syndrome with other psychopathological formations have been studied on the basis of clinical and follow-up examination of 314 patients, 257 male, 57 female, aged 15-17 years old, with phobic disorders diagnosed according to ICD-10 criteria. Simple phobias found in 21% of patients existed as an isolated phenomenon displaying no comorbidity. Asthenic syndrome was most frequently concomitant with phobias (293 patients); personality anomalies were found in 167 patients including 31 with personality disorders; affective syndrome was observed in 61 patients and other ones were noticeably rare. Within the structure of the acute states, phobias appear as a schizophrenic attack or reactive symptoms correlating with the basic syndrome by the time of onset, intensity and content. Age differences of the phobic and related psychopathological syndromes are revealed. PMID- 18427454 TI - [Craniospinal electromagnetic therapy in the neurological practice]. AB - Effectiveness of a new method for treating neurological disorders based on the use of trans-cranial electro- and magnetic stimulation in patients with vascular, vertebral and traumatic pathology of the nervous system is presented. Different frequency parameters of transcranial electrical stimulation and superposition methods of the magnetic generator are proposed taking into account the specific features of neurodynamics and pathogenesis of diseases. The effect of craniospinal electromagnetic therapy on the autonomic nervous system is demonstrated. PMID- 18427455 TI - [The study on the assessment of the new levodopa drug--stalevo (levodopa/carbidopa/entacapone), in treatment of Parkinson's disease in out patient clinical practice (the results of the open START-M)]. AB - Despite significant symptomatic effect of levodopa, in most patients the stable 24 hour's effect changed for alterations of movement symptoms (fluctuations, dyskinesia) after 2-3 years of the therapy. These complications may be reduced due to the constant (physiologic) stimulation of post-synaptic dopamine receptors that, to a certain degree, provides the drug stalevo, which contains levodopa and 2 enzyme inhibitors, dopa decarboxylase (carbidopa) and catechol-O methyltransferase (entacapone). The results of the study demonstrated beneficial effects of stalevo as compared to traditional therapy in patients with "wearing off" and "on-off" phenomena. PMID- 18427456 TI - [Efficacy and safety of galantamine (reminyl) in the treatment of dementia in patients with Parkinson's disease (open-label controlled trial)]. AB - Forty-one Parkinson's disease patients with dementia (21 galantamine group, 20- control group) with onset of dementia at least two years after the manifestation of parkinsonian symptoms participated in this open-label controlled trial of galantamine in maximum dose 16 mg/day. Cognitive, psychiatric and motor symptoms were assessed before and after 4, 12 and 24 weeks of treatment using clinical assessment as well as rating scales, including the Mini-Mental State Examination (MMSE), ADAS-cog, clock drawing test, Frontal Assessment Battery (FAB), and the Neuropsychiatric Inventory (NPI-12) with assessment of caregiver distress. Patients treated with galantamine had better scores on MMSE (p<0,05), ADAS-cog (p<0,05), clock drawing test (p<0,05) and FAB (p<0,01) to the end of the trial comparing to the control group. NPI scores on individual items changed from baseline at week 12 and 24, showing benefits of galantamine treatment as compared to the controls, with significant difference for hallucinations (p=0,0002), anxiety (p=0,04), sleep disorders (p=0,04) and apathy (p=0,006). Galantamine therapy was associated with a significant reduction in caregiver distress (p=0,007), improvement of daily life activity (p=0,003). Gait, freezing and falls were improved in the galantamine group but a mild worsening of tremor was noted in two patients. Adverse events (drooling, postural hypotension, nausea, dysuria) were observed in 7 (30%) of galantamine treated patients. PMID- 18427457 TI - [Pantogam in daily psychiatric practice]. AB - Pantogam was used as a corrector of the side-effects of neuroleptic therapy. The adverse effects were assessed psychometrically with several scales. Attention was focused on the unwanted signs (pathological movements of different groups of muscles). Fifty patients were studied, 37 of them completed the treatment course. Pantogam was administered in dosage 750-3000 mg per day along with traditional and atypical neuroleptics. The drug was effective against the most prevalent side effects. The mild sedative effect of pantogam, absence of negative influence on myocardial vessels and possibility of its combination with most pharmacological drugs make its possible to use the drug in patients of different age groups. PMID- 18427458 TI - [The use of topamax in the treatment of epilepsy: clinical and pharmacoeconomic aspects]. AB - The results of an open prospective naturalistic study on the use of topiramate (topamax) in 92 patients with focal epilepsy are presented. The pharmacological remission with duration over one year was achieved in 47,8% of patients. In total, the percentage of remission and clinical improvement made up 68,5%. The positive changes were observed in drug-naive patients (during first monotherapy) and in patients who previously received ineffective therapy. In some cases of patients with resistant forms of epilepsy, pharmacological remission or significant improvement of the control over seizures was found. The topamax therapy substantially increased the quality of life of the patients. The cost of therapy was 3100-5200 rubles per month, mean costs for a one patient during one year--416,000 rubles. The marginal utility 237,000 rubles per each additional QALY was significantly lower than $20,000, the minimal threshold of cost effectiveness for European countries. PMID- 18427459 TI - [Magnetophoresis of glutaminic acid and magnesium sulfate in rehabilitation of children with cerebral palsy]. AB - Clinical efficacy of magnetophoresis of glutaminic acid and magnesium sulfate used in the rehabilitative complex for preschool children with spastic forms of cerebral palsy has been studied. The clinical and neurophysiological examination has been conducted in 40 children aged from 1 to 7 years. The higher efficacy of treatment with combination of running pulsed magnetic field and drugs as compared to magnetotherapy using the same procedure has been found. PMID- 18427460 TI - [Delta-sleep inducing peptide and the drug deltaran: possible approaches to antistress protection]. AB - An aim of the present study was a comparative investigation of a delta-sleep inducing peptide and the drug deltaran on the neural activity of the brain structures involved in emotional processing. Another goal was to analyze the possibility to prevent negative effects of emotional stress on brain ischemia using, along with deltaran, glycine and a delta-sleep inducing peptide. Deltaran and the delta-sleep inducing peptide exert in general similar effect on the burst activity of neurons in the dorsal hippocampus, hypothalamic paraventricular nucleus and ventral anterior thalamic nucleus, inducing amplification of the majority of recorded units. The activation of neuronal activity was seen mostly after the delta-sleep inducing peptide microiontophoresis in the dorsal hippocampus and after the deltaran application in the hypothalamic paraventricular nuclei. The index characterizing blood supply was significantly higher in all rats receiving deltaran as compared to the controls. Animals receiving deltaran survived experimental brain ischemia in 100% cases versus 38% in those not exposed to this drug. PMID- 18427462 TI - [Decreased numerical density of pericapillary oligodendrocytes in the cortex in schizophrenia]. AB - Autopsy samples from the prefrontal cortex were obtained from 22 schizophrenic cases and 22 normal controls. Capillaries and oligodendrocytes were viewed in paraffin sections stained with a Luxol-fast blue + cresyl violet. Electron microscopy was applied to study the ultrastructure of oligodendrocytes. Numerical density of pericapillary oligodendrocytes was measured in layer V of Broadmann's area 10. Subjects with schizophrenia had a significantly lower (-20,8%) numerical density of pericapillary oligodendrocytes compared to controls. The analysis for schizophrenic subgroups with predominantly positive or predominantly negative symptoms revealed a significant decrease in the numerical density of pericapillary oligodendrocytes only in the latter one (-29,4%). The numerical density of pericapillary oligodendrocytes in cases with predominantly positive symptoms also decreased (-8,3%) but not differed significantly from the controls. Electron microscopy revealed the prominent ultrastructural dystrophic and degenerative alterations of pericapillary oligodendrocytes in the prefrontal cortex in schizophrenic brains. The present study points out that the population of pericapillary oligodendrocytes is involved in the pathogenesis of schizophrenia. Damage and loss of pericapillary oligodendrocytes might lead to disruption of the blood-brain barrier and as a result to dysregulation of metabolic trafficking across the blood-brain barrier in the prefrontal cortex in schizophrenia. PMID- 18427461 TI - [The role of antibodies to phospholipids in the pathogenesis of symptomatic epilepsy]. AB - Antibodies to phospholipids (ABPL) were studied in 40 patients, aged from 17 to 65 years, with symptomatic epilepsy. Those with primary or secondary antiphospholipid syndrome and with diffuse diseases of connecting tissues were not included in the study. All patients underwent clinical, neurological, electroencephalographic and immunological examination, the latter including the determination of ABPL level, namely the level of AB to cardiolipin in the blood serum using ELISA. It was shown that the ABPL level was 1,5-2,0 times higher in patients comparing to the controls. This increase was observed in patients with moderate and high frequency of seizures and a history of at least 2 seizures. Moreover, the ABPL level was depended on the etiologic factor and epileptic EEG activity. In conclusion, the results obtained suggest that ABPL not only interact with vessel endothelium but caused the immunologically mediated damage of cortical neurons that plays an important role in the pathogenesis of symptomatic epilepsy. PMID- 18427463 TI - [Treatment of gastroduodenal bleedings: from surgery to therapy?]. AB - Results of treatment of 2352 patients with gastroduodenal ulcer bleedings were analyzed. Surgical treatment was performed at 495 (21%) of them. Emergency life saving operation was performed at 78 (15%) patients, urgent operation after primary hemostasis - at 114 (23%), elective operations - at 303 (62%). Various resections were carried out at 78% cases, organ-saving surgeries - at 22%. The main problem in the treatment program of these patients is the prophylaxis of bleeding recurrence. Rational pathogenetic out-patient treatment of ulcer disease, prophylaxis of recurrences and complications are the key points which need close collaboration between surgeons and gastroenterologists. PMID- 18427464 TI - [Multivisceral resections at disseminated abdominal tumors]. AB - Overall 131 multivisceral resections at disseminated abdominal malignant tumors have been performed. The main arguments to extended operations were local dissemination of tumor (35.1%), synchronous hepatic metastases (49.6%), combination of local dissemination with synchronous metastases (6.1%), isolated synchronous tumors (6.1%), and multiple metastases with polyorganic localization (3%). The rate of postoperative complications and lethality at the patients undergone multivisceral operations were 34.3 and 6.8% respectively. There were no significant differences in postoperative complications rate and lethality between adjusted multivisceral and standard operations groups. PMID- 18427465 TI - [Comparative analysis of long-term results of combined and surgical treatment at the patients with signet ring cell carcinoma of stomach]. AB - Overall 194 patients with signet ring cell carcinoma of stomach were treated, 124 (64%) of them underwent combined treatment (preoperative radiotherapy with subsequent operation), 70 (36%) - only surgical treatment. There were no differences in survival rate between two groups at early (pT1) cancer. The combined method has advantages over isolated surgical at T2 tumors (both with and without regional lymphatic metastases) and at T3 tumors without regional lymphatic metastases (p=0.03). At T3N1-2 tumors with regional lymphatic metastases and T4 tumor neither combined no surgical method demonstrated positive results of treatment. Generally combined treatment with preoperative radiotherapy has statistically significant advantage (p=0.007) over surgical treatment at the patient with signet ring cell carcinoma of stomach. PMID- 18427466 TI - [Local permanent intraarterial antimicrobic therapy in postoperative period at the patients with cholangitis]. AB - Method of local permanent intraarterial antimicrobic therapy was used in postoperative period at the patients with acute cholangitis. The method includes transfemoral catheterization of hepatic artery and permanent infusion of ceftriabol (2.0 ml per day) and metronidazol (1.0 ml per day) during 5-7 days. At the end of first week the more positive results were achieved compared with control group. PMID- 18427467 TI - [Prophylaxis of pulmonary complications after traditional cholecystectomy at senior patients]. AB - Results of complex treatment of 238 patients undergone operations due to cholelithiasis with acute cholecystitis are analyzed; 198 of them were aged 60 years. At perioperative period patients were treated traditionally, 76 patients were additionally treated with alkaline inhalations, bronchodilatators, cardiac drugs, Bemithyl antihypoxant, broad spectrum antibiotics, oxygen therapy through nasopharyngeal catheter and oxygen cocktail, physical exercise therapy and prolonged lung ventilation (in postoperative period). Pulmonary complications were registered in 2.5 times more at traditional treatment in senior patients compared with ones less than 60 years. Original complex treatment permitted to reduce the rate of postoperative complications in 2.96 times. PMID- 18427468 TI - [Pathogenesis and prophylaxis of venous thromboembolic complications at large ventral hernias]. AB - Overall 188 patients undergone surgical treatment due to large postoperative ventral hernias are analyzed. It is demonstrated that venous hemodynamics at these patients before surgery is characterized by disorders of "abdominal-caval pump" function due to anterior abdominal wall destruction and low mobility of diaphragm's cupula. The recovery of their functions is the key point in prophylaxis of thromboembolic complications. PMID- 18427469 TI - [Surgical treatment of atherosclerotic lower limb ischemia with ulcers and necrotic lesions at the patients aged 70 years and older]. AB - Overall 133 patients aged 70 years and older with critical lower limb ischemia and necrotic lesions were treated. It is demonstrated that arterial reconstructive surgery is the effective method for limb salvage, life prolongation and improvement of life quality. PMID- 18427470 TI - [Methodology of clinical studies in surgery]. AB - Results of clinical studies recently became the guidelines in decision making in medicine including surgical practice. Results of each study could be used generally only if all methodological requirements are fulfilled. In this paper the principles of clinical studies design particularly in surgery and medical publications analysis are described and discussed. PMID- 18427471 TI - [Classification of acute inflammation of pancreas]. AB - The common classifications of acute pancreatitis are analyzed. The original classification based on experience of the treatment more 3000 patients is suggested. This classification reflects ethiology, pathogenesis, severity, clinical manifestation and complications of pancreatitis and permits to optimize the treatment of patients. PMID- 18427472 TI - [Endoscopic ultrasonography in diagnosis of surgical treatment of pancreas]. AB - Results of endoscopic ultrasonography at 137 patients suspected for pancreas pathology are analyzed. Methodology of endoscopic ultrasonography, semiotics of pancreas surgical diseases, advantages over other diagnostic methods are described. Endosonography is informative method for final diagnosis of different disease of pancreas. Diagnostic value of method and area of its clinical application are described. PMID- 18427473 TI - [Peculiarities of saliva chemiluminescence and microelement hair content in children with various dental caries activity]. AB - Integrated stomatological examination of 1002 children of Khabarovsk Territory was conducted. Besides carious process and oral hygiene indices the intensity of free radical oxidation in saliva (by chemiluminescence method) and basic microelement hair content (by spectral analysis method) in urban and rural children with compensated and decompensated forms of dental caries. High and equal prevalence and incidence of dental caries in urban and rural children was found. It was shown that in cases of decompensated forms of dental caries indicators of saliva chemiluminescence and consequently the intensity of free radical oxidation was significantly higher than in cases of compensated forms of dental caries. It was determined that activation of free radical oxidation with dental caries progression was developed on the background of combined deficit and imbalance of essential microelements. PMID- 18427474 TI - [Histamine releasing activity of dental materials as the indicator of their biocompatibility]. AB - Different types of dental materials (DM) were studied for their capacity to release histamine in vitro from basophils of whole blood of allergic patients and healthy donors using automated and computerized glass fiber-based leukocyte histamine release test (LHRT). It was shown that some types of DM possessed ability to release histamine from basophils and some didn't. There were no differences in histamine releaseability from basophils obtained from allergic patients and healthy donors. LHRT gives opportunity to recognize of DM possessing high or low histamine releaseability as well as to detect individual sensitivity to different DM. Thus, LHRT can be used for preliminary assessment of DM for their biocompatibility and also for individual selection of suitable DM for particular patient to avoid unwanted side effects. PMID- 18427475 TI - [Experimental substantiation of indirect electrochemical blood oxidation and antioxidant therapy use efficacy in the treatment of festering soft tissues wounds]. AB - In experiment on rats in the treatment of festering soft tissues wounds it was substantiated the inclusion into the scheme of the basic therapy 0.03% solution of sodium hypochlorite and antioxidant Recsod. Introduction of these preparations let to reduce the level of oxidation stress and endogenous intoxication, to eliminate imbalance in the antioxidant erythrocyte system and in the organism as a whole in the acute inflammation phase and to speed up wound healing processes. PMID- 18427476 TI - [Evaluation of the efficacy of porous alumina ceramic use for defects elimination in maxillofacial region]. AB - To study the biocompatibility of porous glass ceramics, experiments were conducted on: 75 white mice (Vistar); 11 rabbits (Shinshila) and 6 dogs. The porous alumina ceramic implants were embedded subperiostaly without additional fixation to bone surface. The osseointegration of implants, the size and form maintaining, changing of physical properties and long-term results in bone substitution were studied. The handling properties during surgery were also evaluated. PMID- 18427477 TI - [Use of TachoComb adhesive material after mucogingival operations]. AB - The most difficult situation is a treatment of, so called, "open" injuries in the oral cavity. A method of treatment of the "open" wound is the transfer it to the "closed" one. Today resolved adhesive materials are used for a covering of the "open" injury. One of the newest materials is TachoComb ("Nycomed", Austria). An effectiveness of using TachoComb in cases of mucogingival operations of patients with somatic diseases was studied. PMID- 18427478 TI - [Periodontal tissues status after treatment of chronic periodontitis with consideration of gender and age]. AB - Radiovisiographic study of periodontal tissues changes in different periods after the therapy of destructive chronic periodontitis have disclosed that jaw alveolar process bone density and the size of the destruction focus after 6 months, 1 and 2 years after the treatment were not significantly changed (134 patients sample of different gender and age). When this sample was subdivided into groups depending upon gender and age some significant changes of some parameters were found. The most expressed were the changes of the size of the periodontal destruction focus in the patients of 40-49 years of age, less pronounced they were in patients of 18-29 age group; in the age group 30-39 years changes were not found, In men of 18-29 age group regenerative processes in periodontal tissues were more expressed than in women. In patients of female sex of the age group 30-39 years were found more expressed changes. In older age group in patients of both genders the changes were approximately equal. The bone density around periodontium after treatment in men of the age group 18-29 years was restored more intensive than in 40-49-year-old patients. In females age dependent changes in the studied tissues status were not found. PMID- 18427479 TI - [Temporo-mandibulary joint state after distraction osteogenesis use in cases of different genesis facial skeleton deformities elimination]. AB - Distraction osteogenesis is one of the most often used methods for the treatment of craniofacial deformities. One of the clinical effects of the distraction is the tissue growth induction process in the conditions of its influence upon condylar process of mandible. Besides it distraction mandible osteosynthesis and active functional therapy stimulates neoarthrosis development in cases of temporo mandibulary joint ankylosis. According to our opinion distraction osteogenesis as the technique for facial skeleton deformities elimination is indicated in cases of surgical treatment of joint ankylosis, deforming arthrosis for mandible size recovery and patients' functional rehabilitation. PMID- 18427480 TI - [Estimation of tracheal intubation methods in patients with pathology of maxillofacial region]. AB - In 42 patients with pathology of face cranium tracheal intubation was performed by different methods--intubation with artificial jet high frequency lung ventilation (I+AJHFV) and fiber optic intubation (FOI). It was established that more favourable conditions for intubation were created when I+AJHFV was used. FOI with preserved consciousness and spontaneous breathing in patients with pathology of maxillofacial region reduces the time factor significance but demands significant sedative preparation of the patient. PMID- 18427481 TI - [Remote results of surgical treatment of aging ptosis of face and neck tissues]. AB - Remote results of surgical treatment of patients with aging ptosis of face and neck tissues during 15 years are presented. Results were good there where suggested incisions marking was carried out and for those face and neck skin regions which should be corrected. Unsatisfactory results were received in the cases when wounds edges were sewn by uninterrupted blanket sutures without skin deeper layers fixing. PMID- 18427482 TI - [Principles of bone tissue structures interaction with full removable dentures fixed on intraosseous implantates modelling]. AB - Math modelling "removable dentures-implantate-bone" with size and density of bone tissue as variables was created. It allowed to study biomechanical bases of mandibular bone tissue structures interaction with full removable dentures of different constructions and fixed on intraosseous implantates. Analysis of the received data showed that in the majority of cases it was expedient to recommend 3 bearing (abutments) system of denture making. Rest on 4 and more implantates was appropriate for patients with reduced density of spongy bone and significant mandibular bone atrophy. 2 abutment system can be used in patients with high density of spongy bone and absence of mandibular bone atrophy. PMID- 18427483 TI - [Results of the use of whole ceramic prosthetic constructions from zirconium oxide for recovery of the teeth with deep subgingival defects]. AB - 30 patients (13 men and 17 women) with deep subgingival defects were selected for treatment, their mean age was equal to 39.7+/-3.2 years; for their teeth recovery 38 stump inlays from zirconium oxide were made. The patients were distributed in 3 groups depending upon the material used for inlay fixation; for 2 years in the dynamic regime (after 6, 12 and 24 months) clinical evaluations were done of the quality of tooth recovery. Medium-term results of the quality of tooth recovery clinical evaluations by inlays from zirconium oxide have shown that this method is perspective for teeth recovery with deep subgingival defects. Such qualities of metal-free ceramics as practicality, biocompatibility, stability and excellent aesthetic characteristics have provided high quality of the treatment. At the same time stability and longevity of the prosthetic constructions were formed not only by mechanic characteristics of the used materials but also by such fixating cements properties as the degree of edge protection and resorption resistance. Adhesive inlay fixation with the use of such cements as Panavia F and Fuji plus have demonstrated stabile joint with perfect edge conformity. PMID- 18427484 TI - [Phenotypical analysis of children with congenital cleft of upper lip and palate]. AB - The relation between congenital cleft of upper lip and palate with dysplasia of connective tissue was studied. 102 patients (aged from 1 to 14 years) with various kinds of congenital cleft of upper lip and palate were observed. The reports showed phenotypical attributes of dysplasia of connective tissue of patients with similar pathology. The data on the character and prevalence of external features are received from children with congenital cleft of upper lip and palate. PMID- 18427485 TI - [Early application of plasmapheresis for complex treatment of purulent mediastinitis]. AB - Acute purulent mediastinitis (APM) is one of the most difficult forms of surgical infection with severe clinical course and lethality from 17 to 80%. Overall 80 patients with APM were treated; plasmapheresis in postoperative period was used at 66 of them. The main criteria of negative prognosis at the patients with APM were elderly age, high score on APACHE-2, SAPS-2 and SOFA scales, higher level of creatinine and urea serum concentration, hyperglycemia, leucopenia. Early application of plasmapheresis was associated with lower lethality (12.5% at the group with early beginning of plasmapheresis, 35.3% at the group with late beginning, 36.2% at the control group). Plasmapheresis promotes normalization of hemostasis, microcirculation and immune parameters, reduction of intoxication and systemic inflammation syndrome. PMID- 18427486 TI - [Use of Amplatzer septal occluder system for atrial septal defect closure]. AB - Short-term results of minimally invasive transcatheter closure of atrial septal defect using Amplatzer septal occluder system (AGA Medical Corporation, USA) at 72 patients are analyzed. Technique of surgical procedure is described in detail. Morphometric protocol used for determination of indications and contraindication for endoscopic closure is also described. The causes of impossibility of endovascular procedures are established. The rate of transoccluder blood bypasses in postoperative period is demonstrated. PMID- 18427487 TI - [Efficacy of endoscopic dissection of perforant veins in lower limb chronic lymphovenous insufficiency]. AB - Overall 156 patients with lower limb chronic lymphovenous insufficiency were treated and analyzed. All the patients were divided into 3 groups depending on method of perforant veins dissection. First group consisted of 46 patients who underwent epifascial dissection (Cokket's operation), 2nd group--50 patients undergone isolated subfascial dissection from small separate incisions (Madden Kim's operation), 3rd group--60 patients after endoscopic subfascial dissection (SEPS technology). The results were followed-up to 5 years. It is demonstrated that SEPS technology is associated with the best results in early and late postoperative period (8.3% of recurrences) and prevents the progress of chronic lymphovenous insufficiency. Subfascial dissection promotes early normalization of lower limb microcirculation parameters. Epifascial dissection permits to achieve satisfactory early results, but long-term results are associated with high rate of recurrences especially at the patients with trophic lesions. PMID- 18427488 TI - [Allotransplantation of testicle]. AB - Allotransplantation of vascularized testicle with mild immune suppression was performed at 132 patients with primary hypogonadism associated with androgenic deficient and endocrine form of impotence. This method permits to achieve stable engraftment in early postoperative period at 83% patients. Acute immunologic rejection crisis and secondary wound disruption are the most frequent complications at early postoperative period. The rate of these complications may be reduced with more intensive immune suppression. At the patients with smooth recovery the testosterone level starts to increase in 1 week after allotransplantation and stabilizes during all first year on the level of normal and subnormal parameters. PMID- 18427489 TI - [Choice of the method of isoperistaltic tubular esophagogastroplasty completion based on intraoperative assessment of vascularization of gastric transplant]. AB - Intraoperative assessment of vascularization of isoperistaltic tubular gastric transplant using angiotensiometry and Doppler flowmetry was performed at 136 patients undergone extirpation of esophagus with esophagogastroplasty. The main criteria of gastric transplant convenience for anastomosis with esophagus were: clear intramural pulse on all circle of gastric transplant by flowmetry data; intramural arterial pressure according to angiotensiometry data have to be not less 80 mm Hg, and the ratio of venous to arterial pressure has not to be more 50%. At inadequate criteria the surgery must be finished with cervical esophago- and gastrostoma. PMID- 18427490 TI - [Treatment tactics of the patients with gastroduodenal ulcer bleeding]. AB - Results of treatment of 263 patients aged 15 to 93 years with gastroduodenal ulcer bleeding are analyzed. There were 200 (76,1%) male and 63 (23,9%) female patients. Gastric ulcers were diagnosed at 85, duodenal ulcers--at 178 patients. Differential approach to conservative and surgical treatment of these complications is described. PMID- 18427491 TI - [Bleedings from ulcers after resection of stomach and vagotomy]. AB - Overall 162 patients with recurrent ulcers after previous operations for ulcer disease were treated. Resection of stomach was performed in 69 (42.6%) of them, vagotomy--in 93 (57.4%). Episodes of bleeding were revealed in anamnesis at 56 (81.2%) patients with peptic ulcers after stomach resection, and at 15 (16.1%)- with ulcers after vagotomy. Bleeding was the cause of urgent hospitalization at 39 (28.1%) patients. One hundred and thirty-six from 162 patients have been operated (including 52 patients after primary resection and 81--after primary vagotomy). Vagotomy at the patients with ulcers after primary resections demonstrates good results. Repeated resection of gastric stump with vagotomy is indicated in great penetrating ulcers. Resection of stomach is the operation of choice in majority cases of recurrent ulcers after primary vagotomy. Stem re vagotomy demonstrates good result an the patients with ulcers of gastro intestinal anastomosis after primary selective proximal vagotomy. PMID- 18427493 TI - [Mini-laparoscopic fundoplication by Nissen in the treatment of gastroesophageal reflux at children]. AB - Results of surgical treatment of 180 children with gastroesophageal reflux are analyzed. Laparoscopic fundoplication by Nissen was used. Positive result was achieved in all the cases, there were no lethal outcomes. Mean time of surgery was 64.4+/-30.2 min, mean hospital stay after operation--4.7+/-1.5 days. It is concluded that laparoscopic fundoplication may be performed at children irrespective of age and body mass, including at the patients with malformations and recurrence of disease. PMID- 18427492 TI - [Complex treatment of the patients with postoperative ventral hernia]. AB - Overall 605 patients with postoperative ventral hernia underwent plasty of anterior abdominal wall by combined methods and on-lay or in-lay disposition of synthetic implant. Concomitant diseases were diagnosed 432 (71.4%) patients that required 495 simultaneous operations at 283 (43.8%) patients. Wound complications after surgery were seen at 21 (3.47%) patients. Long-term results were followed up to 11 years: recurrences of hernia were diagnosed at 12 (1.9%) patients, 3 (0.5%) patients died due to pulmonary embolism. It is concluded that the treatment of patients with postoperative ventral hernia requires complex approach and leads to good short- and long-term results. PMID- 18427494 TI - [Treatment tactics in intussusception in children]. AB - Results of treatment of 235 patients with intussusception are analyzed. The treatment tactics in different period of disease is described in details, the role of echography is emphasized. For the first time the conception of functional (reversible) intussusception was offered, treatment tactics were determined. Results of treatment were studied, conservative and surgical tactics of treatment were substantiated. PMID- 18427495 TI - [Ozone in the treatment of purulent surgical diseases of fingers and hand]. AB - Results of treatment of 116 patients were analyzed retrospectively. Study group consisted of 56 patients who treated with ozonized salt solution (3.3-4 mg/l) administered intravenously and through drainage system; 60 patients (control group) were treated traditionally. Bacteriological, cytological and special biochemical examinations were carried out. It is concluded that combination of ozone therapy with active surgical tactics leads to earlier recovery, early elimination of microorganisms, decreases of endotoxicosis parameters, and reduce hospital stay. PMID- 18427496 TI - [Magnetic resonance imaging in the diagnosis of acute pancreatitis]. AB - Overall 52 patients with acute pancreatitis were examined with MRI and spiral CT. The examination was performed during 24 hours from admission time. Results of examination were assessed confidentially by 3 experts with 5-scores scale. Non parametric chi-square criterion was used for significances detection. MRI demonstrated advantages over spiral CT in diagnosis of fat necrosis, sequestration and hemorrhages. MRI compares well with contrast CT in diagnosis of pancreonecrosis. MRI permits to diagnose destructive lesions in pancreas without contrast agent, and accordingly without additional effect on pancreas and kidneys. It is concluded that MRI is high informative method in diagnosis of acute pancreatitis and its complications. PMID- 18427497 TI - [Depression in Alzheimer's disease]. AB - To study clinical and pathogenetic aspects of depression in Alzheimer's disease (AD), 65 patients with AD have been studied. The higher frequency of depressive symptoms has been revealed in different types of AD and all stages of the disease. Different syndrome variants of depression have been found: anxious (35,4%), apathic (29,2%), depression with delusions (16,9%), hypochondriac (10,8%) and melancholic (7,7%). The variety of clinico-phychopathological signs and mechanisms of depressive pathology formation in patients with AD suggests its heterogenic character. PMID- 18427499 TI - [Pelotherapy of patients with diabetic poloyneuropathy: a clinical-immunological study]. AB - Clinical presentations, stage of disease, therapeutic efficacy and parameters of humoral and cell immunity were studied in 91 patients with diabetic poloyneuropathy (DP) receiving pelotherapy. It was shown that changes both in cellular and humoral links were increased depending on DP stage. Pelotherapy led to the regress of basic objective and subjective manifestations of DP in all stages of disease. The immunocorrective effect of the treatment was revealed. PMID- 18427498 TI - [Specific disturbances of psychomotor development in children with thymomegaly]. AB - Ninety children, aged from 2 month to 3 years, with thymomegaly and 25 aged matched controls were studied. Most children with thymomegaly had disturbances of psychomotor development. Depending on their types, the cohort of children was stratified into 4 subgroups: 1st - 36 patients (40%) with schizotypal signs; 2nd 30 hyperactive children (33%); 3rd - 19 children with hyperthymia signs (21%); 4th - 5 normal children (6%). The deviations of locomotion and psychiatric development were correlated with the extent of thymus enlargement and activation of innate and adaptive immunity. PMID- 18427500 TI - [Implication of alpha-lipoic acid preparations in the treatment of diabetic neuropathy]. AB - Twenty-eight patients, 17 women and 11 men, aged 40-70 years, with distal sensory motor diabetic neuropathy of the lower extremities were treated with berlition, an alpha-lipoic acid preparation, in dosage 600 mg daily during 3 months. The treatment resulted in the significant reduction of clinical (sensory and motor) and neurological changes of lower extremity peripheral nerves. PMID- 18427501 TI - [Dynamics of biochemical parameters in patients with anxious depression treated with serotonergic antidepressants with different mechanisms of action]. AB - The aim of the study was to compare the dynamics of some biochemical parameters in patients with anxious depression treated with tianeptine (coaxil) - serotonin reuptake enhancer and sertraline (zofort) - selective serotonin reuptake inhibitor. Forty-three patients with ICD-10 diagnosis of anxious depression (F32.1 and F33.1) were included in the study. Comparing to healthy volunteers, the significant increase of platelet monoamine oxidase activity by 95% and concentration of middle molecules (middle-mass endotoxic molecules) by 86% and decrease of serum amine oxidase activity by 43% and functional albumin activity by 38% was found in patients. An analysis of the biochemical profiles revealed the changes in opposite directions of all investigated parameters for antidepressants with different effects on serotonin reuptake though both tianeptine and sertraline were effective in the treatment of anxious depression. PMID- 18427502 TI - [The complex neurochemical assessment of brain proteins in mentally healthy subjects and schizophrenic patients]. AB - Relative amounts of the glutamate metabolizing enzymes - glutamine synthetase, glutamine synthetase-like protein, three isoenzymes of glutamate dehydrogenase as well as creatine phosphokinase (a main astroglial energy metabolism enzyme) and major proteins of astro- and oligodendroglia - a glial fibrillary acidic protein and a myelin basic protein were determined in postmortem brain extracts from three areas - the prefrontal cortex, caudate nucleus and cerebellum - from mentally healthy subjects (n=21) and patients with chronic schizophrenia (n=23). To single out "metabolic types" the data obtained have been subjected to cluster analysis. It has been demonstrated for the first time that the cluster analysis of the biological parameters (enzymes and proteins) with correction for age, gender, postmortem interval and presence/absence of diagnosis, enables to distinguish "mentally healthy" cases and "schizophrenic patients" with a high degree of significance (mean mixing error <20%, small er, Cyrillic>>0,00004). Thus, we suppose that mentally healthy controls and patients with schizophrenia are objectively divided into different "metabolic types". PMID- 18427503 TI - [A nasal cycle in healthy children]. AB - A nasal cycle was studied in 10 healthy children (5 boys and 5 girls) aged 11-17 years (mean age 14.1+/-1.9 years). Registration of the nasal cycle was conducted for 7 hours by means of long-term nasal monitoring with Rhinocycle rhinoflowmeter (Interacoustics, Denmark). The nasal cycle was presented as a graphic image of fluctuations of a relative volumetric flux (RVF) in the relative units in accordance with linear dependence of pressure in the air flow to its volume. The nasal cycle in all healthy children was irregular, with mean fluctuation rate 1.3+/-0.6 h. There were no significant differences by RVF between the right and left parts of the nose in 8 children. In girls the nasal cycle most often was partially coinsiding, partially fluctuating while in most boys (in 4 of 5) the nasal cycle was irregular bilateral and fluctuating. It was found that the cycle was not longer or regular with the child's growth. PMID- 18427504 TI - [Hearing in children with a history of acute otitis media. Use of paracentesis]. AB - The examination of 103 children aged 1 year to 5 years had acute otitis media purulenta. They were treated with paracentesis which had a positive effect on recovery of the acoustic function. It is shown that children with previous acute otitis media should be followed up to control recovery of hearing. PMID- 18427505 TI - [Specific features of mineral and protein metabolism in patients with paranasal osteoma]. AB - A comparative analysis of biochemical blood picture was made in 20 patients with osteoma of the paranasal sinuses before and after surgical treatment. A total of 17 tests characterizing metabolism and 680 biochemical examinations were made. Investigation of mineral and other kinds of metabolism may be important in diagnosis of osteomas, hyperostosis, exostosis and malignant tumors, determination of activity and tumor growth rate. PMID- 18427506 TI - [Diagnostic and expert value of functional tests in detection of latent vestibular dysfunction]. AB - Vestibular dysfunction was studied in 150 patients with paroxysmal states and minimal neurological defect in the past. A series of functional stress tests proved to be highly effective in diagnosis of latent vestibular dysfunction. PMID- 18427507 TI - [Epidemiological features of noninvasive pneumococcal infections in different age groups of population]. AB - The aim of the study was to determine nosological structure, sex and age of patients with non-invasive pneumococcal infections (otitis and sinusitis). Medical cards of 80 inpatients and results of clinico-microbiological tests have been analysed. Otitis and sinusitis morbidity is about 12+/-0.54 and 36+/-1.04 per 10 000, respectively. Acute otitis occurred in 47.5%, acute sinusitis or exacerbation of chronic sinusitis - in 52.5% (S. pneumoniae was isolated). Males suffered more frequently (57.9%) from otitis, females - from sinusitis (54.76%). Registration of pneumococcal otitis and sinusitis morbidity is important for organization of anti-epidemic measures. PMID- 18427508 TI - [Endoscopic surgical treatment of vascular tumors of the paranasal sinuses and base of the skull]. AB - Fourteen endonasal endoscopic operations for vascular tumors of the paranasal sinuses, mainly of stage II-III, were performed in 1997-2006. In one case intraoperative navigation was used. Preoperative angiography was made in 4 patients with a definite diagnosis of hemangioma. Out of them, in three cases preoperative embolisation of the afferent vessel was conducted; apparent vascular bed in tumor stroma was not found. Follow-up from 1 to 8 years registered no further growth of the tumor in patients operated endoscopically. PMID- 18427509 TI - [CT diagnosis of polypous rhinosinusitis]. AB - Patients with diseases of the nasal cavity and paranasal sinuses (PNS) were examined with computed tomography (CT) with subsequent estimation of the area of the ethmoid bone sinuses (EBS) using Contour program. CT detected the following changes: total area of EBS in polypous rhinosinusitis (PRS) was 12-26 cm2 versus 42-51 cm2 in healthy subjects. In ostiomeatal complex pathology total cell area was 30-41 cm2 while area of at least one EBS group was subnormal and corresponded to sinus area in PRS. Conduction of PNS CT with subsequent calculation of EBS area detects patients with defective structure of the ostiomeatal complex predisposed to development of PRS and allows early surgical correction of anomalous group of EBS. PMID- 18427510 TI - [Potentialities of autofluorescent spectroscopy in diagnosis of laryngeal and pharyngeal tumors]. AB - Laryngeal and pharyngeal mucosa of 50 patients with malignant (n=56%) and benign (n=44%) laryngeal and pharyngeal tumors was examined with autofluorescent spectroscopy using nitrogen laser LGI-505 (337,1 nm). It was found that autofluorescent spectrum of malignant tumors is significantly less intensive than relevant spectrum of healthy mucosa and benign tumors. PMID- 18427511 TI - [Stapedoplasty: one technology, two types of protheses]. AB - The article presents postoperative results of stapedoplasty depending on the type of prosthesis (Teflon or titan). The technique of the operation was the same but snapping the prosthesis around a long process of the incus in application of titan prostheses. Thickness of tephlon prostheses was 0.8-1.0 mm, titan - 0.6-0.8 mm, the length of both prostheses was adjusted individually from 3.5 to 4.25 mm. Efficacy of the prostheses was assessed in 0.125-2 kHz and 3-8 kHz tests. PMID- 18427512 TI - [Monotherapy of rhinitis of various etiology]. AB - The article describes various forms of rhinitis and principles of their management; results of monotherapy of patients with different forms of acute rhinitis (viral, allergic) using homeopathic drug corisalium. PMID- 18427513 TI - [Videoendolaryngeal surgery in primary local laryngeal amyloidosis with involvement of the upper third of the trachea: case report]. AB - Laryngeal amyloidosis is a rare pathology (0.5-1% of all benign laryngeal newgrowths). Five patients with local laryngeal and tracheal amyloidosis were successfully diagnosed and treated endoscopically in the endoscopic department of the P.A. Herzen Moscow Research Cancer Institute in 1987-2007. Videoendolaryngeal operations were performed with application of Nd:YAG laser, argon-plasma coagulation and their combination. The patients were followed up for 15 years maximum. Two most interesting cases of primary local laryngeal amyloidosis with involvement of the upper third of the trachea are presented. Foreign and domestic literature data on diagnosis and treatment of respiratory amyloidosis are reviewed. PMID- 18427514 TI - [Pseudopolymelia and disorders of body scheme]. AB - Two cases of pseudopolymelia are described. In the first case, a feeling of having one more left hand developed in a female patient after the ablation of an arteriovenous malformation of the splenium of the corpus callosum expended to the left cingulated gyrus. No movement and sensory disorders were found but "split brain" symptoms were observed. To the best of author's knowledge, this is a first description of such a case. In another female patient, the same feeling appeared, along with deafferent pain in the hand, due to tearing off the primary trunks of brachial plexus after the car accident. An analysis of literature and own data revealed that pseudopolymelia is observed in right-handers in the lesion of the deep right-hemisphere regions and accompanied by movement and sensory disturbances. Moreover, the feeling of phantom extremities appears in the lesion of the peripheral nervous system (deafferent phantoms), amputation of extremity and, what is most important, agenesia. Such findings raise a question on genetically determined engram of body scheme. The appearance of left-sided phenomena: pseudopolymelia, unilateral spatial ignorance, phenomenon of "strange hand", after the lesion of the right hemisphere in right-handers only as well as the corpus callosum argue for the common underlying mechanisms. The author suggests that they are based not only on the disturbances of analysis and synthesis of sensory information and movements (realized by the right hemisphere) but also on the defect of consciousness of stimuli realized by the left hemisphere, which can be connected to through the corpus callosum. PMID- 18427515 TI - [Neuropsychological syndromes of infancy]. AB - Neuropsychological syndromes of infancy, using a method by O.V. Bazhenova (1986), were studied in 77 infants with mild perinatal encephalopathy and dysfunction of one brain hemisphere. The right hemisphere syndrome is characterized by the reduction of reactivity and immediate emotional communication, accelerated development of complicated modes of communication and interest in item operations. The left hemisphere syndrome is characterized by the high reactivity to stimulation with generalized activity and weakness of regulation processes, retardation of complicated modes of communication and item operations but the well development of immediate emotional communication. These peculiarities in infants with the left hemisphere syndrome aggravate in the excess of impressions and stimulation and in those with the right hemisphere syndrome - in their limitation. It is suggested that the syndromes are underlie by disturbances of global neurodynamics. PMID- 18427516 TI - [Dynamics of bulimia disorders in anorexia nervosa and bulimia nervosa]. AB - Thirty-three female patients with anorexia nervosa (AN) with bulimia disorders and 54 with bulimia nervosa (BN) were studied. The analysis of clinical presentations and dynamics of bulimia disorders in these groups revealed the similarity of types of <> behavior. At the same time, NA differs by the absence of genuine binge attacks characteristic of the patients with BN. Psychopathological basis of the <> behavior in AN seems to be dysmorphophobic, anxiety, hypochondriac disorders as well as insufficient criticism of a patient's state in the absence of pronounced affective disorders and reduced self-esteem and that in BN - cyclothymic, bipolar affective disorders and disorders of drive, dysmorphophobic concerns related to the changes of self esteem. Also, patients with BN have criticism towards their behavior. These differences should be taken into account in the choice of therapeutic tactics and methods as well as in rehabilitation of patients with eating disorders. PMID- 18427517 TI - [Serum immunological markers as predictors of effectiveness of antipsychotic therapy in patients with schizophrenia]. AB - Serum immunological parameters - activity of leukocyte elastase (LE) and a1 proteinase inhibitor (a1-PI), content of C-reactive protein, von Willebrand factor, interleukin 8 as well as a level of autoantibodies to neuroantigens (nerve growth factor and basic myelin protein) were studied in patients with schizophrenia during their treatment with psychotropic drugs. All parameters studied differed in the groups of patients and controls. However, the pronounced dynamics was found only for LE and a1-PI activity. After the therapeutic course, the reduction of LE activity accompanied by the increase of a1-PI activity was observed. The correlation study between the biological parameters during disease exacerbation and therapeutic effectiveness assessed by the PANSS scores revealed that activity of LE and a1-PI may be considered as a prognostic marker of therapeutic efficacy, i.e. the high enzyme activity at the moment of maximal activity of the process was predictive of good therapeutic response. PMID- 18427518 TI - [The use of coaxil (tianeptine) in the treatment of aged people with combined mild cognitive impairment and depressive-anxiety disorders]. AB - Twenty patients, aged 60-69 years, with combined mild cognitive impairment, subtle depression and anxiety have been treated with coaxil (tianeptine) used in dosage 37,5 mg daily during 60 days. The clinical state of patients has been assessed using clinical-psychopathological examination as well as the Hamilton scale and neuropsychological testing according to A.R. Luria. The high effectiveness of coaxil is revealed with respect to all psychopathological characteristics. Recommendations on expediency of administering coaxil to aged patients with combined cognitive and depressive disorders are presented. PMID- 18427519 TI - [Assessment of berlition effectiveness in ischemic stroke with CT perfusion imaging of the brain]. AB - Berlition, an alpha-lipoic acid preparation, is a natural regulator of the free radical processes. It was used as an add-on therapy in the management of patients in the acute period of ischemic stroke during 10 days. A main group included 15 patients treated with berlition and a comparison group - 11 patients who received a basic therapy. Patients' age was 30-60 years. A complete clinical study of patients was conducted during 21 days of treatment. CT perfusion imaging of the brain with contrast material was carried out on 1st, 5th and 15th days. The higher therapeutic effectiveness was found in the main group of patients, with complete functional rehabilitation in 33% of cases and significant regress of neurologic symptoms in 67%. These changes were correlated with positive shifts of different hemodynamic parameters to the 15th day of treatment. PMID- 18427520 TI - [Diagnostics and treatment tactics for chronic cerebrovascular disorders with the use of single photon emission computed tomography]. AB - Single photon emission computed tomography (SPECT) is a highly informative method of functional neuroimaging with high diagnostic potential for some neurological disorders. We used this method in diagnostics of specification of lesion localization, choosing of treatment tactics for chronic cerebral blood circulation disorders and evaluation of cavinton effect. The positive effect of the drug on cerebral hemoperfusion is confirmed. PMID- 18427521 TI - [Natural neurotrophic IgG autoantibodies in the blood serum of patients with different clinical outcomes after ischemic stroke]. AB - Recent studies revealed a role of natural neurotrophic IgG autoantibodies (NNAA) in the trophic supply of nervous tissue and modification ability of neuronal networks. Changes of the NNAA level likely may reflect direction (compensation, decompensation) of cytochemical and morphologic alterations of nervous tissue in acute focal ischemia, thus, having certain prognostic value. To assess the dynamics of NNAA in the blood serum on 3, 7, 14 and 21 days after ischemic stroke and 3 months after the disease, 84 patients (mean age 68+/-10 years) with first ever stroke in the system of internal carotid artery were studied. NNAA immunoreactivity to four nervous tissue proteins, S100b, GRAP, MP-65, NGF, was conducted using ELIZA method. Patients' outcomes were assessed with the Barthel Index and NIH on the 3rd month of the disease. In the acute period of stroke, the changes of immunoreactivity, comparing to controls, were found in 95% o cases (p<0,01), the level of NNAA pathologically increased or decreased. The increase of NNAA level was revealed in 45 patients (56% cases) with satisfactory clinical rehabilitation on 7-14 days (p<0,0001) with the following normalization of these parameters to the 21st day. In case of unfavorable outcomes, the prolonged increase of the NNAA level or decrease of immunoreactivity (p<0,0001) was found in 44% of cases. The changes of NNAA level were observed in 60 patients on the 3rd month of the disease, i.e. the protracted decrease of immunoreactivity was found in 42 patients and increase - in 18 cases (p<0,01). The quantitative changes of NNAA likely indicate the adaptive-compensative reaction of the immune system. The character of immunoreactivity changes is related to reparative neuroplasticity and may be of prognostic value. PMID- 18427522 TI - [Neurophysiological abnormalities in the P300 paradigm as endophenotypes of schizophrenia]. AB - The aim of the study was to discriminate the auditory event-related potentials (ERP) abnormalities in the P300 paradigm as the possible neurophysiological endophenotypes (pathogenetic predisposition markers) of schizophrenia. Deviations of the parameters of different waves of auditory ERP were determined in a sample of patients as compared to the controls, the influence of patient's clinical state (measured by PANSS) upon ERP abnormalities was studied. The comparison of neurophysiological characteristics was carried out also in mentally healthy parents from the families with so-called "sporadic" cases of schizophrenia and in parents from the families, in which the other spouse was either affected with mental disorder or had relatives with schizophrenia. The data obtained suggest that two neurophysiological abnormalities of auditory ERP - the decrease of N100 amplitude in ERP to non-target stimuli and reduction of P300 amplitude, fulfill the "endophenotype" criteria. PMID- 18427523 TI - [Extirpation of stomach without esophagoenteroanastomosis at recurrent profuse gastric bleedings]. AB - Five cases of extirpation of stomach stump are analyzed. These operations have been performed due to recurrent bleeding after long conservative treatment, endoscopic and surgical procedures (gastrotomy with suturing of erosions and ulcers, resection and re-resection of stomach) at the patients with severe anemia and hypoproteinemia. The operations were limited to extirpation without esophagoenteroanastomosis due to severe condition of patients and were completed by external esophageal drainage and jejunostoma. Three of five patients have been survived and undergone reconstructive operations with Roux esophageal-intestinal anastomosis in 4 - 6 months. All these patients resumed professional activity. We didn't find the similar surgical tactics in literature. PMID- 18427524 TI - [Algorithm of cardiac risk assessment before vascular surgery]. AB - THE OBJECT: to detect the informative value of algorithm of cardiac risk assessment recommended by American Heart Association compared with original standard enlarged protocol. Overall 456 patients underwent vascular operations. First group consisted of 198 patients who were examined according original enlarged protocol; other 258 patients (2nd group) were examined with AHA protocol. At 1st group 41% patients didn't need myocardial revascularization (according coronarography results), preventive coronary bypass surgery was performed at 32% patients. At 2nd group coronarography was performed at 133 (51%) patients according risk factors stratification and stress-examination results. Preventive coronary bypass surgery was performed at 97 patients; 27 patients refused this recommendation. Enlarged protocol increases in 2.6 times the odds of unnecessary invasive procedure. It is concluded that AHA algorithm permits to reduce in 2 times and more the number of invasive diagnostic procedures compared with standard enlarged protocol; AHA algorithm increases the clinical and economic effectiveness. PMID- 18427525 TI - [Staging treatment of aorta aneurysm disease]. AB - Overall 13 patients with aorta aneurysm disease underwent staging operations with 2 weeks - 12 years interval. Surgical treatment of aneurismal lesion of the whole of aorta is very difficult due to severity of the disease and non-typical surgical technique, and characterized by high complications rate and lethality. Individual approach to each patients and staging surgery permitted to achieve the positive results of surgical treatment with 10% lethality. PMID- 18427526 TI - [Prediction of critical lower limb ischemia at the patients with lower limb arteries atherosclerosis]. AB - Arterial inflow, venous outflow and microcirculation were studied with ultrasound Doppler, photopletismography and oxygen partial pressure detection at 115 patients with lower limb arteries atherosclerosis and different stages of limb ischemia. Four-level algorithm of critical limb ischemia prediction has been created. First level is the study of arterial inflow. Gradient of regional systolic blood pressure >or=2.0 is the critical parameter. Second level is the study of venous outflow. Critical parameter - gradient of postocclusive venous pressure >or=3.0. Third level is the ratio of arterial and venous blood flow. Venous-arterial index >or=40% demonstrates critical disturbances of macrohemodynamics. Fourth level is the study of microcirculation. If the capillary gradient 50%) of anxiety are reported in patients undergoing screening for prostate cancer, which may affect health-related quality of life. We aimed to determine the level and prevalence of anxiety and depression and to identify those aspects of the diagnostic pathway that induce the most stress in men being investigated for prostate cancer. A total of 159 prostate-specific antigen unscreened men undergoing a transrectal ultrasound-guided biopsy of the prostate (TRUS-B) completed two questionnaires, prior to their biopsy and before receiving results, containing the Hospital Anxiety and Depression Scale (HADS) and a 10 point Visual Analogue Scale (VAS). Median scores and prevalence of anxiety (4-5, 4-7%) and depression (1-2, 1.4%) respectively were low for both questionnaires. Waiting for biopsy results received the highest median VAS score (6) and was the most stressful event in 65% of men. There is a low incidence of clinically significant anxiety and depression in men being investigated for prostate cancer but questionnaires such as HADS identify patients with psychological distress who may benefit from early counselling. Uncertainty about the future while awaiting biopsy results after TRUS-B seems to be the most stressful event in patients' lives and minimizing this wait should help optimize patient care. PMID- 18427570 TI - Race and prostate weight as independent predictors for biochemical recurrence after radical prostatectomy. AB - We hypothesized that factors beyond pathological stage, grade, PSA and margin status would be important predictors of biochemical recurrence (BCR) after radical prostatectomy (RP). A cohort of 3194 patients who underwent RP between 1988 and 2007 and who had neither neoadjuvant therapy nor postoperative adjuvant hormonal therapy was retrieved from the Duke Prostate Center database. Age, prostate-specific antigen (PSA), pathological Gleason score (pG), lymph node status, seminal vesicle invasion (SVI), extracapsular extension (ECE), positive surgical margin (PSM) status, year of surgery, race, adjuvant radiation therapy (XRT), percent tumor involvement in the RP specimen and prostate weight were evaluated as possible predictors of BCR in multivariate Cox regression analysis. BCR was defined as a PSA of 0.2 ng ml(-1) or higher at least 30 days after surgery. A nomogram was developed from the Cox model. Predictive accuracy was obtained by calculating bias-corrected Harrell's c and by bootstrap calibration. In multivariate analysis, PSA (hazard ratio 1.39 (95% confidence interval 1.29 1.51)), ECE (1.22 (1.04-1.44)), pG score (1.38 (1.14-1.68), 2.23 (1.76-2.84), 2.69 (2.12-3.40) for pG 3+4, 4+3, >7, respectively), SVI (1.72 (1.40-2.12)), PSM (2.05 (1.73-2.42)), year of surgery (0.65 (0.54-0.77)), African-American race (1.37 (1.13-1.66)), adjuvant XRT (0.19 (0.11-0.34)) and prostate weight (0.83 (0.76-0.92)) were identified as independent predictors of BCR (P< or =0.018 for all factors). Predictive accuracy of the nomogram was 0.75. Race and prostate weight were independent predictors for BCR after RP. By incorporating these variables, we developed a nomogram, which provides a highly accurate means for estimating risk of BCR after RP. PMID- 18427571 TI - Biochemical relapse-free survival in 400 patients treated with I-125 prostate brachytherapy: the Guildford experience. AB - A total of 1200 patients had undergone I-125 prostate brachytherapy (BXT) in our centre. We present prospective outcome data for the first 400 treated patients. Data were analysed from a prospective database of 400 consecutive patients treated with permanent prostate BXT between March 1999 and December 2003. Patients were stratified into low (49%), intermediate (36%) and high (15%) risk as defined by the Memorial Sloan-Kettering Prognostic Index. Patients received 145 Gy BXT alone (41%), BXT with 3 months neoadjuvant androgen deprivation (NAAD) (39%), 45 Gy external beam radiotherapy (EBRT) with 110 Gy BXT (3%) or a combination of NAAD, 45 Gy EBRT and 110 Gy BXT (17%). Biochemical relapse-free survival (bRFS) and prostate-specific antigen (PSA) nadirs were analysed for treatment received in each risk group. Median follow-up was 54 months (range, 38 96 months) with a mean patient age of 63 years. Prostate cancer-specific survival was 99.5%. Twenty-eight patients (7%) experienced biochemical failure according to the 2006 Radiation Therapy Oncology Group-American Society for Therapeutic Radiology and Oncology (RTOG-ASTRO) Phoenix Consensus definition (PSA nadir plus >or=2 ng ml(-1)): nine low-, fourteen intermediate- and five high-risk patients. When stratified by treatment group for low-, intermediate- and high-risk groups, the 5-year actuarial bRFS was 98, 89 and 100% for BXT; 91, 87 and 88% for NAAD and BXT; 100, 80 and 100% for EBRT and BXT; and 100, 92 and 88% for NAAD, EBRT and BXT, respectively. Overall 4- and 5-year PSA or =3 lines, or percentage > or =20/40. CONCLUSIONS: Removal of the ILM decreases persistent and recurrent macular holes. ICG staining and complete removal of the ILM slows the rate of visual recovery but does not appear to have any long-term deleterious effect on the results of macular hole surgery. PMID- 18427611 TI - Central corneal opacification resulting from recent chemotherapy in corneal donors. AB - PURPOSE: Ocular surface disease following penetrating keratoplasty has been shown to increase patient morbidity and adversely affect graft survival. Five cases of dense central subepithelial opacification were noted in keratoplasty patients who received tissue from donors who had chemotherapy prior to death. Cancer-related deaths account for approximately 20% of the cornea donor pool. The purpose of this study was to identify the effect of recent systemic antimetabolite therapy on donor corneas. METHODS: Eye bank donor charts of 120 consecutive penetrating keratoplasty donors were retrospectively reviewed for cancer-related deaths. Donors who received chemotherapy prior to death were identified. Recipient records of those patients receiving tissue from donors that had recently undergone systemic chemotherapy were reviewed. Corneal clarity and postoperative ocular surface disease were noted by the surgeon. RESULTS: Twenty-nine of 120 cornea donors (24%) had a cancer-related cause of death. Five of these 29 donors (17%) had undergone systemic chemotherapy with antimetabolite drugs (which inhibit microtubule formation) within the previous 8 weeks. All 5 recipients postoperatively developed central subepithelial opacification in spite of temporary tarsorrhaphy performed at the time of keratoplasty. Central corneal opacification was severe enough in 3 patients to require regrafting for visual improvement. Histopathology showed abnormality of the central epithelial basement membrane, apoptosis of basal epithelial cells, and thinning of the epithelial cell layer. CONCLUSION: Corneas from donors who received a full course of recent systemic antimetabolite therapy are associated with central subepithelial scarring following keratoplasty. Corneal surgeons should be aware of the potential for severe ocular surface disease when using donor tissue from patients with metastatic cancer. PMID- 18427612 TI - How effectively does medical care achieve its purposes? Evaluation of peer reviewed literature in ophthalmology related to wellness. AB - PURPOSE: To get an idea of whether the issue of what makes people healthier is studied in ophthalmology by determining the proportion of articles dealing with that subject. METHODS: Prospective review of all articles published in 3 consecutive issues of 7 peer-reviewed ophthalmology journals, using a grading system in which A signified an article that clearly dealt with a subject expected to have an impact on health or quality of life, or that considered health or quality of life itself directly; B indicated an article similar to A, but not directly concerned with the issue of health; C signified an article similar to B but more distantly related to health or quality of life; and D was the grade given when there was no relationship at all to health or quality of life. Grading was done independently by 3 graders. A literature review on the subject was also performed. RESULTS: Thirty-three articles received a grade of A, 229 of B, 740 of C, and 81 of D. There were more articles that had no relationship at all to health or quality of life than there were articles dealing directly with those issues. CONCLUSIONS: On the basis of a review of the literature and of over 1000 articles, ophthalmologists do not appear to give much priority to issues of quality of life or health. How validly these conclusions can be generalized to general clinicians is not known. PMID- 18427613 TI - Aqueous humor dynamics in monkeys in response to the kappa opioid agonist bremazocine. AB - PURPOSE: To determine the effects of the kappa opioid agonist, bremazocine (BRE), on intraocular pressure (IOP) and aqueous humor dynamics in normotensive cynomolgus monkeys. METHODS: IOP, pupil diameter, refraction, aqueous humor flow, and mean arterial pressure (MAP) were measured following unilateral topical application of 1 to 100 microg BRE. IOP and MAP responses to 100 microg BRE were repeated during intravenous infusion of angiotensin II (ATII). IOP and MAP responses to BRE were also measured following pretreatment with the opioid receptor antagonists norbinaltorphimine (nor-BNI) or naloxone. Outflow facility was measured following unilateral intracameral exchange with 0.01 to 100 microg/mL BRE. IOP, aqueous humor flow, pupil, and MAP were measured after unilateral intracameral bolus injection of 1 microg of BRE. RESULTS: Unilateral topical BRE caused a dose-related reduction in IOP and aqueous humor flow in both eyes and in MAP. Pupil miosis occurred at the 100-microg dose. There was no effect on refraction. IOP and MAP decreases after 100 microg of BRE were eliminated by ATII infusion. Differential IOP effects after 10-microg topical BRE doses were not eliminated by nor-BNI or naloxone. Unilateral intracameral bolus injection of BRE decreased IOP in both eyes but had no effect on MAP or aqueous humor flow. Outflow facility was unchanged after intracameral exchange with BRE. CONCLUSIONS: The IOP response to high doses of BRE in monkeys can be attributed to peripheral or central effects on MAP. The IOP-lowering response to topical BRE is due to aqueous humor flow suppression via non-opioid receptor stimulation. Some components of the IOP response are mediated by unknown mechanisms. PMID- 18427614 TI - Topographically guided LASIK for myopia using the Nidek CXII customized aspheric treatment zone (CATz). AB - PURPOSE: To assess the efficacy, predictability, and safety of topography-guided laser in situ keratomileusis (LASIK) for the surgical correction of low to moderate myopia with astigmatism using the Nidek CXIII excimer laser equipped with the customized aspheric treatment zone (CATz) algorithm. METHODS: In a multicenter US Food and Drug Administration study of topography-guided LASIK, 4 centers enrolled 135 eyes with manifest refraction sphere that ranged from -0.50 to -7.00 D (mean, -3.57 +/- 1.45) with up to -4.00 D of astigmatism (mean, -1.02 +/- 0.64 D). The intended outcome was plano in all eyes. Refractive outcomes and higher-order aberrations were analyzed preoperatively and postoperatively. Patient satisfaction was assessed using both the validated Refractive Status and Vision Profile (RSVP) questionnaire and a questionnaire designed for this study. Six-month postoperative outcomes are reported here. RESULTS: By 6 months postoperatively, the manifest refraction spherical equivalent (MRSE) for all eyes was -0.09 +/- 0.31 D. Six months postoperatively, 116 of 131 eyes (88.55%) had an uncorrected visual acuity of 20/20 or better, and 122 of 131 eyes (93.13%) had a MRSE within +/-0.50 D. Distance best spectacle-corrected visual acuity (BSCVA) increased by 2 or more lines in 21 of 131 eyes (19.01%), and no eyes lost 2 lines or more of BSCVA. The total ocular higher-order aberrations root-mean-square increased by 0.04 microm postoperatively. Patients reported significantly fewer night driving and glare and halo symptoms postoperatively than preoperatively. CONCLUSIONS: Nidek CXIII CATz treatment of myopia with astigmatism is safe, efficacious, and predictable, and it reduces patient symptoms associated with night driving and glare and halo symptoms. PMID- 18427616 TI - Monocular diplopia due to spherocylindrical refractive errors (an American Ophthalmological Society thesis). AB - PURPOSE: Ordinary spherocylindrical refractive errors have been recognized as a cause of monocular diplopia for over a century, yet explanation of this phenomenon using geometrical optics has remained problematic. This study tests the hypothesis that the diffraction theory treatment of refractive errors will provide a more satisfactory explanation of monocular diplopia. METHODS: Diffraction theory calculations were carried out for modulation transfer functions, point spread functions, and line spread functions under conditions of defocus, astigmatism, and mixed spherocylindrical refractive errors. Defocused photographs of inked and projected black lines were made to demonstrate the predicted consequences of the theoretical calculations. RESULTS: For certain amounts of defocus, line spread functions resulting from spherical defocus are predicted to have a bimodal intensity distribution that could provide the basis for diplopia with line targets. Multimodal intensity distributions are predicted in point spread functions and provide a basis for diplopia or polyopia of point targets under conditions of astigmatism. The predicted doubling effect is evident in defocused photographs of black lines, but the effect is not as robust as the subjective experience of monocular diplopia. CONCLUSIONS: Monocular diplopia due to ordinary refractive errors can be predicted from diffraction theory. Higher order aberrations--such as spherical aberration--are not necessary but may, under some circumstances, enhance the features of monocular diplopia. The physical basis for monocular diplopia is relatively subtle, and enhancement by neural processing is probably needed to account for the robustness of the percept. PMID- 18427617 TI - Dissociated horizontal deviation: clinical spectrum, pathogenesis, evolutionary underpinnings, diagnosis, treatment, and potential role in the development of infantile esotropia (an American Ophthalmological Society thesis). AB - PURPOSE: To elucidate the pathophysiology of dissociated horizontal deviation. METHODS: The reversed fixation test was performed prospectively in 28 patients who developed consecutive exotropia following horizontal extraocular muscle surgery for infantile esotropia. All patients were assessed for the presence of adduction weakness, latent nystagmus, dissociated vertical divergence, and neurologic disease. RESULTS: A positive reversed fixation test, indicating the presence of dissociated horizontal deviation, was found in 14 of 28 patients (50%) with consecutive exotropia. In patients with dissociated horizontal deviation, the exodeviation was usually smaller with the nonpreferred eye fixating than with the preferred eye fixating, and smaller with the preferred eye fixating than during periods of visual inattention or under general anesthesia. Dissociated horizontal deviation correlated with the findings of dissociated vertical divergence, but not with asymmetric adduction weakness, latent nystagmus, or neurologic disease. CONCLUSIONS: Using reversed fixation testing, dissociated horizontal deviation can be detected in 50% of patients who develop consecutive exotropia following surgery for infantile esotropia. In this setting, monocular fixation with either eye superimposes a dissociated esotonus upon a baseline exodeviation. Fixation with the nonpreferred eye usually exerts greater esotonus than fixation with the preferred eye, producing an asymmetrical exodeviation during prism and alternate cover testing. Depending on the baseline anatomical position of the eyes, this dissociated esotonus can manifest as an intermittent exodeviation or an intermittent esodeviation. This unrecognized form of ocular motor dissociation may contribute to the pathogenesis of infantile esotropia. PMID- 18427618 TI - Hanging by a thread: the long-term efficacy and safety of transscleral sutured intraocular lenses in children (an American Ophthalmological Society thesis). AB - PURPOSE: To evaluate the long-term efficacy, safety, and advisability of using transscleral sutured posterior chamber intraocular lenses (IOLs) in pediatric patients with no capsular support and to determine whether 10-0 polypropylene suture should be used for this purpose. METHODS: A long-term retrospective interventional case series review of 33 eyes of 26 patients who had a sutured IOL at Duke University Eye Center were evaluated for the intraoperative surgical risks, postoperative visual and refractive outcomes, and the number, type, and severity of the postoperative complications. In addition, a survey of pediatric ophthalmologists' experience with suture breakage was performed. RESULTS: Postoperative visual acuity was significantly improved after surgery (P < .001). Predicted vs actual refraction was not significantly different (P = .10) and was within 1.50 diopters of predicted in 66% of patients. A refractive myopic shift occurred over time and was age-dependent. Intraoperative and immediate postoperative complications were minimal and not sight-threatening. Three patients developed subluxation of the IOL secondary to spontaneous 10-0 polypropylene suture breakage at 3.5, 8, and 9 years after surgery. A survey of pediatric ophthalmologists revealed 10 similar cases (mean, 5 years after surgery). CONCLUSION: Transscleral fixation of an IOL in a child appears to be a safe and effective procedure provided that the suture material used is stable enough to resist significant degradation over time. Caution should be exercised in the use of 10-0 polypropylene suture to fixate an IOL to the sclera in children, and an alternative material or size should be considered. PMID- 18427621 TI - A reconsideration of anterior chamber flare and its clinical relevance for children with chronic anterior uveitis (an American Ophthalmological Society thesis). AB - PURPOSE: To gain a better understanding of laser flare photometry values ("flare") as a feature of chronic anterior uveitis in children; to identify relationships between flare and other patient and disease characteristics; to describe changes in flare during course of disease; and specifically to determine whether elevated flare is predictive of subsequent adverse events. METHODS: A retrospective review of medical records was performed for all children (aged < or =16 years at disease onset) with chronic anterior uveitis presumed to be noninfectious, who were examined by one clinician at the Jules Stein Eye Institute since laser flare photometry became available at that facility. All involved eyes were studied. Cross-sectional analysis compared initial flare to other characteristics. Relationships between potential risk factors and outcomes were studied by Kaplan-Meier analyses and Cox proportional hazards regression models. RESULTS: Included were 114 patients (198 involved eyes). Follow-up ranged from 0 to 154.8 months (median, 23.5 months for 82 patients with follow-up). Flare was related to the following factors: anterior chamber cells, keratic precipitates, papillitis, and various complications of uveitis, including band keratopathy, posterior synechiae, and cataract. Flare was not a function of disease duration. High flare was associated with an increased risk of vision loss and development of new vision-threatening complications, including glaucoma/increased intraocular pressure, during follow-up; risk was independent of anterior chamber cells. CONCLUSIONS: Flare may be useful in the evaluation and management of chronic anterior uveitis in children. Flare is a marker of disease severity and is predictive of adverse events during the course of disease. PMID- 18427620 TI - Comparison of topical interleukin-1 vs tumor necrosis factor-alpha blockade with corticosteroid therapy on murine corneal inflammation, neovascularization, and transplant survival (an American Ophthalmological Society thesis). AB - PURPOSE: Interleukin-1 (IL-1) and tumor necrosis factor-alpha (TNF-alpha) play critical roles in mediating corneal inflammation. In this study, topical blockade of IL-1 and TNF-alpha, alone or in combination, was compared to conventional corticosteroid anti-inflammatory therapy in suppressing infiltration of the cornea by antigen-presenting Langerhans cells (LCs) and in promoting corneal transplant survival in a mouse model of keratoplasty. METHODS: Study drugs included topical 2% IL-1 receptor antagonist (IL-1Ra), 1.5% soluble TNF-alpha receptor (sTNFR), and 1% prednisolone phosphate (Pred), all formulated in hyaluronic acid vehicle. Fifty eyes of BALB/c mice were used for LC studies where the numbers of LCs were determined 1 week after electrocautery to the corneal surface or transplantation of C57BL/6 corneas. Additionally, 65 BALB/c mice received corneal allografts and were randomized to receive one of the following for 8 weeks: (1) IL-1Ra, (2) sTNFR, (3) Pred, (4) combined IL-1Ra and Pred, or (5) vehicle alone. RESULTS: Mean suppression of LC infiltration after electrocautery or transplantation was 67% and 71%, respectively, for IL-1Ra, 40% and 62% for sTNFR, 70% and 72% for sTNFR+IL-1Ra, and 77% and 78% for Pred alone. Rejection rates were 15% for IL-1Ra (P = .01), 38% for sTNFR (P = .1), 17% for Pred (P = .02), and 7% for combined IL-1Ra+Pred (P = .002) as compared to 69% for the vehicle-treated group. IL-1Ra and Pred, but not sTNFR, significantly inhibited post-transplantation neovascularization. CONCLUSIONS: Topical IL-1Ra and prednisolone are comparable in their capacity to promote graft survival. sTNFR therapy, though effective, has much lower efficacy as compared to IL-1Ra or Pred. Combination IL-1Ra and steroid therapy offers only minimal added efficacy over either agent used alone. PMID- 18427619 TI - Sources of binocular suprathreshold visual field loss in a cohort of older women being followed for risk of falls (an American Ophthalmological Society thesis). AB - PURPOSE: To determine the sources of binocular visual field loss most strongly associated with falls in a cohort of older women. METHODS: In the Study of Osteoporotic Fractures, women with severe binocular visual field loss had an increased risk of two or more falls during the 12 months following the eye examination. The lens and fundus photographs of the 422 women with severe binocular visual field loss, plus a random sample of 141 white women with no, mild, or moderate binocular visual field loss--47 white women with no binocular visual field loss, 46 white women with mild binocular visual field loss, and 48 white women with moderate binocular visual field loss--were evaluated for lens opacities, glaucomatous optic nerve damage, age-related macular degeneration, and diabetic retinopathy. RESULTS: Eighty-four percent of the women with severe binocular visual field loss had ocular disease in one or both eyes. Bilateral cataracts and glaucomatous optic nerve damage were the most common sources of this severe binocular visual field loss. Approximately 15.2% of women had no evidence of lens opacities, glaucomatous optic nerve damage, age-related macular degeneration, or diabetic retinopathy. CONCLUSION: Severe binocular visual field loss due primarily to cataracts, glaucoma, and age-related macular degeneration explains 33.3% of the falls among women who fell frequently. Because binocular visual field loss may be treatable and/or preventable, screening programs for binocular visual field loss and subsequent referral for intervention and treatment are recommended as a strategy for preventing falls among the elderly. PMID- 18427622 TI - Suppression of keratoepithelin and myocilin by small interfering RNA (an American Ophthalmological Society thesis). AB - PURPOSE: Mutations of keratoepithelin (KE) and myocilin (MYOC) have been linked to certain types of inherited corneal stromal dystrophy and open-angle glaucoma, respectively. In this study, the feasibility of using small interfering RNAs (siRNAs) to suppress the expression of keratoepithelin and myocilin and their capabilities to reduce the related cytotoxic effects caused by mutant myocilins were investigated. METHODS: cDNAs of human KE gene and myocilin gene were amplified by polymerase chain reaction and subcloned into pEGFP-N1 to construct respective plasmids, KEpEGFP and MYOCpEGFP, to produce fluorescence-generating fusion proteins. Short hairpin RNAs (shRNAs) were generated from an RNA polymerase III promoter-driven vector (pH1-RNA). Transformed HEK293 and trabecular meshwork (TM) cells were cotransfected via liposomes with either KEpEGFP or MYOCpEGFP and respective shRNA-generating plasmids to evaluate the suppression efficacy of shRNAs. Suppression of KE-EGFP by KE-specific shRNAs was evaluated by fluorescence microscopy and Western blotting. Suppression of MYOC EGFP by myocilin-specific shRNAs was quantified with UN-SCAN-IT software on digitized protein bands of Western blots. A BiP promoter-driven luciferase reporter assay was used to evaluate the stress response of TM cells induced by misfolded mutant myocilins. RESULTS: Two KE-specific shRNAs that effectively suppressed the expression of KE-EGFP in HEK293 cells were identified. One shRNA (targeting the coding sequence starting at 1528bp of KE) reduced the expression of KE-EGFP approximately by 50%, whereas the other shRNA (targeting the 3'-UTR region of KE) suppressed greater than 80% of the expression. Cotransfection of MYOCpEGFP and various shRNA-generating plasmids targeting different regions of myocilin (containing amino acid residues R76, E352, K423, or N480 associated with inherited glaucoma) showed effective reduction of MYOC-EGFP, ranging from 78% to 90% on average. The activation of BiP gene (as a stress response induced by mutant myocilins) in transformed TM cells was significantly reduced when mutant myocilin proteins were suppressed by myocilin-specific shRNAs. CONCLUSIONS: KE- or myocilin-specific shRNAs could effectively suppress the expression of recombinant KE or myocilin proteins and the related cytotoxicity of mutant myocilins. RNA interference may have future therapeutic implications in suppressing these genes. PMID- 18427623 TI - Mechanisms of retinal ganglion specific-cell death in Leber hereditary optic neuropathy. AB - PURPOSE: Leber hereditary optic neuropathy (LHON) results from point mutations in mitochondrial DNA (mtDNA) present in all cells but is only manifested in retinal ganglion cells (RGCs). Given that RGCs use superoxide for intracellular signaling after axotomy, and that LHON mutations increase superoxide levels in non-RGC transmitochondrial cybrids, I hypothesized that RGCs regulate superoxide levels differently than other neuronal cells. METHODS: Superoxide production in mitochondria isolated from the RGC-5 cell line, rat brain, or neuroblastoma SK-N AS cells was measured and correlated with levels of mitochondrial electron transport chain (METC) complexes. RESULTS: The rate of superoxide production in brain mitochondria was more than 5 times the rate in RGC-5 cells when complex I substrates were used. Rotenone significantly increased the rate of superoxide production in brain but not RGC-5 mitochondria. Succinate-dependent superoxide production was similar in brain and RGC-5 mitochondria, but was increased by the complex III inhibitor antimycin A only in brain cells. Neuroblastoma mitochondria demonstrated similar superoxide generation rates as brain cells. Lower rates of superoxide production probably reflected lower levels of METC components. CONCLUSIONS: These results demonstrate that RGC-5 mitochondria produce superoxide at significantly lower rates than brain mitochondria. Tighter regulation of superoxide levels in RGCs would prevent aberrant apoptosis signaling. LHON mtDNA mutations may interfere with superoxide regulation, possibly leading to aberrant RGC death and consequent optic neuropathy. PMID- 18427625 TI - Impact of a pay-for-performance intervention: financial analysis of a pilot program implementation and implications for ophthalmology (an American Ophthalmological Society thesis). AB - PURPOSE: To determine whether a specific pay-for-performance program design will result in a decrease in global health care expenditures attributable to implementation of that program. METHODS: A retrospective analysis was performed of costs referable to the health plan during a baseline year in comparison to the year following the program implementation. All claims paid during the year prior to program implementation (Baseline) were compared with all costs during the first year of program deployment (Intervention). The primary outcome measure was global health plan expenditure. Secondary outcomes measures included global health plan expenditures adjusted for catastrophic cases and changes in costs by provider type attributable to the program implementation. RESULTS: Global expenditures, for Implementation relative to Baseline years, decreased to $2,049,780 from $2,316,929 (11.5%). When adjustment was made for catastrophic cases, costs decreased to $1,645,568 from $1,811,840 (9.2%). This cost reduction was achieved despite approximately a 10% increase in provider pricing per unit of service. CONCLUSIONS: In this pilot, implementing the program was an effective way to reduce the total health care costs in the first year of implementation. This supports the concept and documents for the first time in a commercial population that an appropriately designed pay-for-performance system can reduce total health care costs by reduction in units of service. This reduction in units of service will more than offset a substantive increase in physician payment per unit of service. Pay-for-performance measures will impact the practice of ophthalmology as government, payers, employers, and consumers focus on value and on demonstrable, auditable outcomes of the care process. PMID- 18427626 TI - Malpractice and the quality of care in retinopathy of prematurity (an American Ophthalmological Society thesis). AB - PURPOSE: A review of retinopathy of prematurity (ROP) malpractice cases will identify specific, repetitive problems in the provision of care and the reasons underlying these problems. Opportunities to improve the quality of care provided to premature infants with ROP will result. METHODS: A retrospective review of a series of 13 ROP malpractice cases in which the author served as a paid consultant, as well as a review of the literature for additional cases, was conducted. The series of 13 involved a review of the entire medical record as well as testimony and depositions. The characteristics of each case are tabulated, including state, date, allegations, defendants, disposition, award, the medical facts and care issues involved, and the judgment of medical error. In addition, a merit review was performed on the care in each case, and an error assessment was performed. RESULTS: The quality of care issues included neonatology failure to refer or follow up in 8 of 13, failure to adequately supervise resident care in 2 of 13, ophthalmologic failure to follow up in 6 of 13, and failure to properly diagnose and manage in 9 of 13. The latter included 4 of 13 that hinged on zone III issues and the presence or absence of full nasal vascularization with or without previous zone II disease. Merit review found negligent error by at least one party in 12 of 13. Ophthalmology error was found in 6 of 13. Malpractice, ie, negligent error causing negligent harm, was judged to be present in 9 of 13. CONCLUSIONS: Negligent errors are common in malpractice cases that proceed to disposition. There are a limited number of repetitive errors that produce malpractice. An explanation of how these errors occur, coupled with the pertinent pathophysiology, afford an excellent opportunity to improve patient care. PMID- 18427624 TI - A prospective study of cavernous sinus surgery for meningiomas and resultant common ophthalmic complications (an American Ophthalmological Society thesis). AB - PURPOSE: Cavernous sinus surgery is considered neurosurgically feasible. A systematic review of patients undergoing cavernous sinus procedures for meningioma was undertaken to determine whether cavernous sinus surgery could be performed with an acceptable level of iatrogenic-induced dysfunction. METHODS: Fifty-six patients undergoing 57 cavernous sinus surgical procedures performed by a single senior neurosurgeon were systematically evaluated to determine the consequences of surgery. Quantitative assessment of afferent (acuity, fields, pupil) and efferent function was stressed. RESULTS: Five of 20 patients (25%) with preoperative optic nerve dysfunction improved, but vision worsened in 6 (30%), including 4 (20%) whose vision deteriorated to no light perception. Four (11%) of 37 patients developed newly acquired optic neuropathy. No patients with preoperative third nerve palsies (19) cleared, although one improved. All 57 patients had evidence of some cranial nerve dysfunction (III, IV, V, or VI) immediately after surgery. Eight patients with long-term follow-up had complete sixth nerve palsies (7 preoperatively), and 4 had complete third nerve dysfunction (none in patients normal preoperatively). Nine (16%) had evidence of aberrant regeneration of the third nerve, and 12 (21%) developed neurotrophic keratitis. CONCLUSIONS: Cavernous sinus surgery results in transient worsening of third, fourth, fifth, and sixth cranial nerve function. Cavernous sinus surgery carries a high risk of worsening ocular motor disorders and producing new ones. Preexisting cranial nerve dysfunction (other than optic nerve) rarely improves. Patients and physicians should be aware of the potential for ophthalmic complications in addition to the more generalized risks of neurosurgery (eg, cerebrospinal fluid leak, infection, stroke). PMID- 18427627 TI - The graded levator hinge procedure for the correction of upper eyelid retraction (an American Ophthalmological Society thesis). AB - PURPOSE: Many surgical techniques have been developed to address eyelid retraction with varying results. Identifying and evaluating the anatomical and pathophysiological factors involved will assist in its surgical treatment. This prospective study evaluated the graded levator hinge procedure, in combination with a Mullerectomy and/or lateral canthoplasty when indicated, in an attempt to precisely and selectively target the pathophysiology responsible for the various causes of eyelid retraction in only one surgical session. METHODS: This is a clinical, prospective study of patients with moderate to severe eyelid retraction due to various causes who underwent the graded levator hinge procedure, in combination with a Mullerectomy and/or lateral canthoplasty when indicated. The exact amount of hinging of the levator aponeurosis, and combination with a Mullerectomy and/or lateral canthoplasty, was determined by the clinical operative findings with active cooperation from the conscious patient. RESULTS: Thirty-two consecutive patients (48 eyelids) with varying degrees of upper eyelid retraction underwent the graded levator hinge procedure in combination with a Mullerectomy and or lateral canthoplasty when indicated. The mean (+/- standard deviation) preoperative palpebral vertical fissure height was 12.4 mm (+/- 0.45 mm), and the mean postoperative palpebral fissure height was 9.0 mm (+/-0.20 mm). The mean preoperative asymmetry in the palpebral fissure height was 2.41 (+/- 0.29) mm, and the mean postoperative asymmetry was 0.59 mm (+/- 0.09), and this difference was statistically significant (P <.001). The mean reduction in the palpebral fissure height was 4.6 mm (+/- 0.29 mm) (range, 1-10 mm). The graded levator hinge procedure in combination with a Mullerectomy and or lateral canthoplasty when indicated, led to a statistically significant (P <.001) reduction in mean palpebral fissure height for all patients, the bilateral subset of patients, the unilateral subset of patients, and the thyroid-related orbitopathy subgroup. The graded levator hinge procedure in combination with a Mullerectomy and/or lateral canthoplasty when indicated led to a statistically significant reduction in palpebral fissure height, asymmetry between the eyes in the total set of patients, the unilateral set of patients, and the thyroid related orbitopathy subset, but not in the bilaterally operated subset of patients, which were already relatively symmetric preoperatively. Postoperatively 90.6 % of all eyelids were within 1 mm of the desired postoperative level (25% were equal, 68.8% were within 0.5 mm, and 6.2% greater than 1 mm from the desired level). CONCLUSIONS: The graded levator hinge procedure, alone or in combination with a Mullerectomy and/or lateral canthoplasty, is a safe and highly effective surgical approach for the treatment of various causes of upper eyelid retraction. Through consideration of the various anatomical and pathophysiological causes of eyelid retraction, excellent functional and cosmetic results are achieved with a graded procedure tapered to the needs of each individual. PMID- 18427628 TI - Preventing surgical confusions in ophthalmology (an American Ophthalmological Society thesis). AB - PURPOSE: Surgical confusions have been rarely studied, especially in ophthalmology. The author hypothesized that such confusions occur rarely but are unacceptable in the public, legal, and regulatory arenas; often occur in circumstances presenting predictable risk; more often involve wrong lens implant than wrong eye, procedure, or patient; and can be prevented by following the Universal Protocol. METHODS: A retrospective series of 106 cases occurring between 1982 and 2005 included 42 closed files from the Ophthalmic Mutual Insurance Company and 64 cases reported to the New York State Health Department. Records were grouped by procedure planned and analyzed to answer these questions: How did the error occur? By whom and when was the error recognized? Who was responsible? Was the patient informed? What was done to the patient? What was the outcome? What liability payments were made? What policy changes or sanctions resulted? Was the error preventable by following the Universal Protocol? RESULTS: The most common confusion was wrong lens implant, accounting for 67 (63%) of the 106 cases. Wrong eye surgery occurred in 15 cases, wrong eye blocks in 14, wrong patient/procedure in 8, and wrong corneal transplant in 2. In 16 cases, the Universal Protocol would have been unlikely to prevent the confusion. CONCLUSIONS: Surgical confusions occur infrequently and usually cause little or no permanent injury, but they may be devastating to the patient, the physician, and the profession. Measures to prevent such confusions, including the Universal Protocol and related checklists, deserve the acceptance, support, and active participation of ophthalmologists. PMID- 18427629 TI - Endothelial keratoplasty: clinical outcomes in the two years following deep lamellar endothelial keratoplasty (an American Ophthalmological Society thesis). AB - PURPOSE: To evaluate the clinical outcome of small-incision, deep lamellar endothelial keratoplasty (DLEK) for the treatment of endothelial dysfunction. METHODS: A prospective series of 79 eyes that underwent DLEK by a single surgeon was evaluated. Best spectacle-corrected visual acuity (BSCVA), refractive astigmatism, and central endothelial cell density (ECD) were measured preoperatively and at 6, 12, and 24 months. RESULTS: Data was available on 78 eyes (99%) at 6 months, 77 eyes (97%) at 1 year, and 79 eyes (100%) at 2 years. Mean BSCVA preoperatively of 20/71 improved to 20/42 by 6 months and remained stable. Eliminating eyes with known retinal disease, BSCVA of 20/40 or better was present in 60% (40 of 67) of eyes at 6 months, 74% (49 of 66) of eyes at 1 year, and 79% (53 of 68) of eyes at 2 years. Refractive astigmatism preoperatively was .91 +/-.78 diopters and was unchanged by surgery over time with results at 6 months of 1.11 +/-.76 (P = .052, power = .43), 1 year 1.04 +/-.80 (P =.287, power = .06), and 2 years 1.10 +/-.70 (P =.467, power = .22). The mean donor ECD preoperatively was 2819 +/- 225 (2389 to 3385) cells/mm(2), and this decreased by 26% at 6 months (2095 +/- 380) (1097 to 2920) (P = .0001; 95% confidence interval [CI] = 643-809), 3% fewer at 1 year (2009 +/- 393) (612 to 2723) (P = .054, power = .5), and 17% fewer at 2 years (1536 +/- 547) (500 to 2546) (P < .001, 95% CI = 368-585). Complications included one primary graft failure and 4 dislocations into the anterior chamber. CONCLUSIONS: DLEK provides improved vision and minimal refractive astigmatic change, but progressive ECD decrease over time is of concern. PMID- 18427631 TI - Computer-assisted quantification of vascular tortuosity in retinopathy of prematurity (an American Ophthalmological Society thesis). AB - PURPOSE: Plus disease is severely abnormal dilation and tortuosity of posterior retinal blood vessels in infants with retinopathy of prematurity (ROP). It has become the major criterion for laser treatment in ROP, but its assessment is subjective and prone to error. ROPtool is a computer program that traces retinal blood vessels, measures their tortuosity, and determines whether there is sufficient tortuosity for plus disease. The purpose of this study was to prospectively determine if assessment of tortuosity by ROPtool is more accurate than by individual pediatric ophthalmologists. METHODS: One hundred eighty-five high-quality RetCam images from premature infants were circulated to 3 ROP experts to develop reference data and to 3 other pediatric ophthalmologists ("examiners") who graded the tortuosity in each quadrant as normal, pre-plus, or plus. These same images were analyzed using ROPtool. Overall accuracy, sensitivity, and specificity of ROPtool relative to expert consensus were compared to that of the individual examiners. RESULTS: By expert consensus, 37 of the 185 eyes (20%) had tortuosity sufficient for plus disease. The overall accuracy of ROPtool of 95% (175/185) for identifying tortuosity sufficient for plus disease was similar to that of examiner 1 (93%, 172/185, P = .5), examiner 2 (93%, 172/185, P = .5), and examiner 3 (91%, 168/185, P = .1). Sensitivity of ROPtool of 97% (36/37) was superior to that of examiner 1 (65%, 24/37, P < .001), examiner 2 (70%, 26/37, P < .001), and examiner 3 (81%, 30/37, P = .02). The mean tortuosity of quadrants with plus disease was 19.1 tortuosity units, compared to 9.9 tortuosity units for quadrants with pre-plus (P < .001) and 4.8 tortuosity units for normal quadrants (P < .001 for pre-plus vs normal). CONCLUSIONS: ROPtool has excellent sensitivity and overall accuracy relative to expert consensus in the detection of tortuosity sufficient for plus disease. Computer assisted analysis of retinal images has the potential to remove subjectivity from the determination of plus disease and to optimize the timing of follow-up and treatment for ROP. PMID- 18427630 TI - Causing and curing infantile esotropia in primates: the role of decorrelated binocular input (an American Ophthalmological Society thesis). AB - PURPOSE: Human infants at greatest risk for esotropia are those who suffer cerebral insults that could decorrelate signals from the 2 eyes during an early critical period of binocular, visuomotor development. The author reared normal infant monkeys, under conditions of binocular decorrelation, to determine if this alone was sufficient to cause esotropia and associated behavioral as well as neuroanatomic deficits. METHODS: Binocular decorrelation was imposed using prism goggles for durations of 3 to 24 weeks (in 6 experimental, 2 control monkeys). Behavioral recordings were obtained, followed by neuroanatomic analysis of ocular dominance columns and binocular, horizontal connections in the striate visual cortex (area V1). RESULTS: Concomitant, constant esotropia developed in each monkey exposed to decorrelation for a duration of 12 to 24 weeks. The severity of ocular motor signs (esotropia-angle; dissociated vertical deviation; latent nystagmus; pursuit/optokinetic tracking asymmetry; fusional vergence deficits), and the loss of V1 binocular connections, increased as a function of decorrelation duration. Stereopsis was deficient and motion visual evoked potentials were asymmetric. Monkeys exposed to decorrelation for 3 weeks showed transient esotropia but regained normal visuomotor behaviors and binocular V1 connections. CONCLUSIONS: Binocular decorrelation is a sufficient cause of infantile esotropia when imposed during a critical period of visuomotor development. The systematic relationship between severity of visuomotor sign, and severity of V1 connectivity deficit, provides a neuroanatomic mechanism for several of these signs. Restoration of binocular fusion and V1 connections, after short durations of decorrelation, helps explain the benefits of early repair in human strabismus. PMID- 18427632 TI - Visual morbidity in thirty-four families with Schnyder crystalline corneal dystrophy (an American Ophthalmological Society thesis). AB - PURPOSE: To assess the findings, visual morbidity, and surgical intervention in Schnyder crystalline corneal dystrophy (SCCD). METHODS: Retrospective case series of 115 affected individuals from 34 SCCD families identified since 1989. Age, uncorrected visual acuity, best-corrected visual acuity (BCVA), corneal findings, and ocular surgery were recorded. Prospective phone, e-mail, or written contact provided updated information. Patients were divided into 3 age categories for statistical analysis: less than 26 years of age, 26 to 39 years of age, and 40 years of age and older. RESULTS: Mean age on initial examination was 38.8 +/- 20.4 (range, 2-81) with follow-up of 55 of 79 (70%) of American patients. While there were no statistical significant correlations between logMAR visual acuity and age (logMAR BCVA =.033 + .002 x age; R =.21), the linear regression showed the trend of worse visual acuity with age. BCVA at > or =40 years was decreased compared to <40 (P < .0001), although mean BCVA was > 20/30 in both groups. Twenty-nine of 115 patients had corneal surgery with 5 phototherapeutic keratectomy (3 patients), and 39 penetrating keratoplasty (PKP) (27 patients). PKP was reported in 20 of 37 (54%) patients > or =50 years and 10 of 13 (77%) of patients > or =70. BCVA 1 year prior to PKP in 15 eyes (9 patients) ranged from 20/25 to 20/400 including 7 eyes with other ocular pathology. BCVA in the remaining 8 eyes was 20/25 to 20/70 with 3 of these 4 patients reporting preoperative glare. Chart and phone survey suggested increasing difficulty with photopic vision with aging. CONCLUSION: Although excellent scotopic vision continues until middle age in SCCD, most patients had PKP by the 7th decade. SCCD causes progressive corneal opacification, which may result in glare and disproportionate loss of photopic vision. PMID- 18427633 TI - The incidence of symptomatic acquired lacrimal outflow obstruction among residents of Olmsted County, Minnesota, 1976-2000 (an American Ophthalmological Society thesis). AB - PURPOSE: To define the incidence, clinical characteristics, and outcomes following treatment of symptomatic acquired lacrimal outflow obstruction (SALOO) in Olmsted County, Minnesota, from 1976 to 2000, and to test the hypothesis that the incidence of this disorder increased over this interval. METHODS: In this retrospective, population-based study, the Rochester Epidemiology Project was used to identify patients above the age of 5 years with SALOO. Patient medical records were reviewed, and the incidence and localization of lacrimal obstruction were determined. RESULTS: Five-hundred eighty-seven patients with SALOO were identified, with an average annual incidence rate of 30.47 per 100,000. Nasolacrimal duct obstruction was most common, with an incidence of 20.24 per 100,000. The increase in incidence from 1976-1979 to 1996-2000 was statistically significant (P=.01). Among 397 patients with nasolacrimal duct obstruction, 107 (27%) were male and 290 (73%) female, with a mean age of 59.5 +/- 22 years. SALOO and nasolacrimal duct obstruction incidence increased with age. Glaucoma, dry eye, cataract, diabetes mellitus, systemic malignancy, cigarette smoking, and hypertension were noted in 5.5%, 8.7%, 37.5%, 10.9%, 18.5%, 26.4%, and 41.1% of patients, respectively. One hundred eleven patients underwent dacryocystorhinostomy, with a success rate of 94.1%. CONCLUSIONS: SALOO incidence increased during the study interval, although a possible plateau effect was noted during the last 5 years of the study period. The majority of patients were female above the age of 66 years. The latter finding, in conjunction with US demographic trends, suggests that the frequency of SALOO may continue to increase in the future. External dacryocystorhinostomy appeared to be effective in the management of nasolacrimal duct obstruction. PMID- 18427634 TI - PCI comes to age as age increasingly comes to PCI. PMID- 18427635 TI - Outcome and quality of life one year after percutaneous coronary interventions in octogenarians. AB - BACKGROUND: While percutaneous coronary intervention (PCI) is increasingly being performed in octogenarians, little is known about the quality of life (QOL) one year after PCI. We assessed the one-year outcome and QOL after PCI. METHODS: Outcome and QOL at one year were assessed in patients of 80 years and older who underwent PCI at our institution. We used the RAND-36 Health Survey to assess health-related QOL at follow-up. The EuroSCORE was used for reference risk assessment. RESULTS: Ninety-eight patients (mean age 82.7+/-2.9 years; 60% female) underwent PCI. Acute PCI was performed in 36% of the patients. Canadian Cardiovascular Society (CCS) angina class before the procedure was class III for 28% and class IV for 64%. Of the patients, 98% were in the highest-risk group (additive EuroSCORE 6+). The overall PCI success rate was 94%. Mortality at one year was 19% (38% acute vs. 12% elective PCI). At followup, general health was rated as fairly good and better then before PCI (CCS I and II: 77%). RAND-36 scores for the mental component were better than scores for the physical component. Physical functioning (41+/-28) and role limitations caused by physical health problems (32+/-37) had the worst scores. The mental component vitality had the lowest (55+/-20) and mental health the highest (70+/-21) score. Social functioning was in general good (67+/-26). CONCLUSION: Octogenarians have a high mortality risk following PCI, especially in acute PCI. In survivors QOL is acceptable with a better mental than physical score. In general, PCI in octogenarians has a positive effect on health perception, with less symptoms of angina pectoris. (Neth Heart J 2008;16:117-22.). PMID- 18427636 TI - Prevalence and misdiagnosis of chronic heart failure in nursing home residents: the role of B-type natriuretic peptides. AB - BACKGROUND/OBJECTIVES: Without knowing the exact CHF prevalence, chronic heart failure (CHF) occurs frequently in elderly people both inside and outside nursing homes. For a diagnosis we have to rely on physical examination and additional tests. We therefore run the risk of missing CHF diagnoses or of diagnosing CHF when we should not. Natriuretic peptide assays have emerged as a diagnostic test but their use in nursing home residents is limited. We examined the number of misdiagnoses, the CHF prevalence and the role of natriuretic peptide. METHOD: Residents in one centre without aphasia, cognitive impairments or metastatic cancer were screened for CHF; the natriuretic peptide levels were measured separately. RESULTS: Of the 150 residents, 103 (64%) were included (79+/-11 years). The diagnosis of CHF was established in 24 of these 103 residents with NTproBNP 1871 (IQR 539 to 4262) and BNP 194 (IQR 92 to 460) pg/ml. A striking result was that of the 24 residents found to have CHF after the screening, 15 (66%) had previously been undetected: NT-proBNP 1146 (interquartile range (IQR) 228 to 3341) and BNP 200 (IQR 107 to 433) pg/ml. Moreover, in 13 out of 22 residents (62%) who had previously been thought to have CHF, the diagnosis was rejected: NT-proBNP 388 (IQR 174 to 719) and BPN 90 (IQR 35 to 128) pg/ml). Regarding the diagnostic accuracy of NT-proBNP and BNP, the optimal cut-off level of NT-proBNP was 450 pg/ml with a sensitivity of 0.71 and specificity of 0.67, and for BNP it was 100 pg/ml with a sensitivity of 0.71 and specificity of 0.70. CONCLUSION: Both undetected and incorrect diagnoses of CHF were common. NT-proBNP and BNP were moderately accurate at diagnosing CHF. CHF prevalence was 23%. (Neth Heart J 2008;16:123-8.). PMID- 18427637 TI - Cardiac remodelling: concentric versus eccentric hypertrophy in strength and endurance athletes. AB - Cardiac remodelling is commonly defined as a physiological or pathological state that may occur after conditions such as myocardial infarction, pressure overload, idiopathic dilated cardiomyopathy or volume overload. When training excessively, the heart develops several myocardial adaptations causing a physiological state of cardiac remodelling. These morphological changes depend on the kind of training and are clinically characterised by modifications in cardiac size and shape due to increased load. Several studies have investigated morphological differences in the athlete's heart between athletes performing strength training and athletes performing endurance training. Endurance training is associated with an increased cardiac output and volume load on the left and right ventricles, causing the endurance-trained heart to generate a mild to moderate dilatation of the left ventricle combined with a mild to moderate increase in left ventricular wall thickness. Strength training is characterised by an elevation of both systolic and diastolic blood pressure. This pressure overload causes an increase in left ventricular wall thickness. This may or may not be accompanied by a slight raise in the left ventricular volume. However, the development of an endurancetrained heart and a strength-trained heart should not be considered an absolute concept. Both forms of training cause specific morphological changes in the heart, dependent on the type of sport. (Neth Heart J 2008;16:129-33.). PMID- 18427638 TI - Multifocal spasm with acetylcholine in Prinzmetal angina. AB - A 60-year-old female patient with Prinzmetal angina and a single non-critical (<50%) focal obstruction in the right coronary artery was referred for percutaneous coronary intervention. Coronary angiography with provocative testing using incremental doses of acetylcholine demonstrated diffuse mild vasoconstriction and multifocal hyperreactive vasoconstriction in apparently normal coronary segments but not at the site of the nonsignificant obstruction. We refrained from intervention and advised avoidance of beta-blockade, and continuation of medical therapy with nitrates and calcium antagonists. (Neth Heart J 2008;16:134-6.). PMID- 18427639 TI - An unexpected finding in a patient with haematuria. PMID- 18427640 TI - Scientific progress - or just another day at the office. AB - Harvey is generally accepted as the patron saint of cardiology. Some shading is not amiss because a medical history canon is by definition an instantaneous photograph. Therefore De Motu Cordis is set in a historic perspective and portrayed as the result of genuine scientific progress. Nevertheless Harvey symbolises the watershed between ancient medicine and modern medicine based on observational and experimental research. We too seem to witness a transitional zone between modern medicine and a new customised medicine, individualised and integrative, firmly based on a further expanding technology. (Neth Heart J 2008;16:143-6.). PMID- 18427641 TI - A word from the dark side. PMID- 18427642 TI - Passing of the editorship to Dr. James Willerson. PMID- 18427644 TI - Dishonesty in medicine revisited. PMID- 18427645 TI - Targeted gene silencing of tumor necrosis factor attenuates the negative inotropic effects of lipopolysaccharide in isolated contracting cardiac myocytes. AB - Bacterial endotoxin (lipopolysaccharide) depresses cardiovascular function; however, the mediators and signaling pathways that are responsible for the negative inotropic effects of lipopolysaccharide are not fully known. We used RNA interference to determine the relative role of tumor necrosis factor with respect to mediating the negative inotropic effects of lipopolysaccharide in isolated cardiac myocytes. Cardiac myocyte cultures were treated with lipopolysaccharide in the presence or absence of small interfering RNAs (siRNA) for tumor necrosis factor. We examined the effects of tumor necrosis factor siRNA on lipopolysaccharide-induced tumor necrosis factor messenger RNA (mRNA) and protein biosynthesis, as well as the negative inotropic effects of lipopolysaccharide in isolated contracting cardiac myocytes. Treatment of adult cardiac myocyte cultures with tumor necrosis factor siRNA significantly attenuated lipopolysaccharide-induced tumor necrosis factor mRNA and protein biosynthesis, whereas transfection with a double-stranded RNA that does not target mammalian mRNA had no effect. Pretreatment with tumor necrosis factor siRNA significantly attenuated, but did not abrogate, the lipopolysaccharide-induced decrease in sarcomere shortening in isolated contracting cardiac myocytes. In contrast, tumor necrosis factor siRNA had a comparatively smaller effect on improving sarcomere shortening once the negative inotropic effects of lipopolysaccharide were fully established. These results suggest that tumor necrosis factor plays an important upstream role in lipopolysaccharide-induced negative inotropic effects in isolated contracting cardiac myocytes and that other molecular mechanisms are responsible for the decrease in sarcomere shortening after sustained lipopolysaccharide signaling. PMID- 18427646 TI - Evaluation of left ventricular long-axis function in cases of rheumatic pure mitral stenosis with atrial fibrillation. AB - Using tissue Doppler echocardiography, we evaluated left ventricular long-axis function in 65 mitral stenosis patients, 30 of whom were in sinus rhythm and 35 of whom were in chronic atrial fibrillation. There were 35 healthy control subjects. Conventional echocardiography was used to evaluate left ventricular diameters, left atrial diameters, left ventricular ejection fractions, and mitral valve areas. Tissue Doppler echocardiography was used to evaluate isovolumetric contraction time (ICT), isovolumetric relaxation time (IRT), ejection time (ET), and peak systolic myocardial velocities. The myocardial performance index was calculated with the formula (ICT+IRT)/ET. Mean and segmental left ventricular IRT and ICT values were significantly longer in mitral stenosis patients than in control subjects, but were similar in the sinus rhythm and atrial fibrillation subgroups. Ejection times were significantly lower in the mitral stenosis with atrial fibrillation group than in the mitral stenosis with sinus rhythm and control groups (P<0.001), but were similar when the sinus rhythm group was compared with the control group (P<0.05). Myocardial performance index values were higher in mitral stenosis patients, significantly so in those with atrial fibrillation when compared with the control group (P<0.001). Systolic velocity values were significantly lower in mitral stenosis patients, more markedly so in those with atrial fibrillation than in the control group (P<0.001). We conclude that patients with pure rheumatic mitral stenosis, most particularly when in combination with atrial fibrillation, have significantly impaired left ventricular long-axis function as evaluated by tissue Doppler echocardiography, although global systolic function may be normal. PMID- 18427647 TI - "Acute takeoff" of the circumflex artery: a newly recognized coronary anatomic variant with potential clinical consequences. AB - Human coronary anatomy allows for a wide range of anatomic variants while maintaining certain consistent features. The use of specific descriptions and names is helpful in talking about variants that have an implicit potential for clinical consequences. In reviewing the angiograms of 813 patients, we newly identified a coronary pattern that we propose to name "acute takeoff of the circumflex artery" in 16 patients (2%). This previously unreported pattern angiographically features a or=220 mg/dl, 5.7 mmol/l) was calculated by adjusting only for age (step 1), for age and lifestyle factors (drinking habit, smoking habit, habitual exercise) (step 2), for age, body mass index (BMI), and the laboratory data [creatinine, glycosylated hemoglobin A1c (HbA1c), aspartic aminotransferase, gamma-glutamyl transpeptidase (gamma-GTP), uric acid] (step 3), and for age, BMI, lifestyle factors, and the laboratory data (step 4). RESULTS: The odds ratio (OR) for shift work with respect to the onset of hypercholesterolemia was significant in step 4 [OR 1.10, 95% confidence interval (95% CI) 1.00-1.21, P-value 0.048], whereas the corresponding odds ratios in steps 1, 2, and 3 were not significant. In step 4, the other significant covariates were age (OR 1.01, 95% CI 1.00-1.02), BMI (OR 1.07, 95% CI 1.05-1.09), HbA1c (OR 1.07, 95% CI 1.02-1.11), gamma-GTP (OR 1.03, 95% CI 1.02-1.04) and alcohol consumption (OR 0.89, 95% CI 0.81-0.99). CONCLUSIONS: In the present study, shift work was shown to be a potential risk factor for hypercholesterolemia among male Japanese workers. However, no consistent association was found between shift work and hypercholesterolemia in the hierarchical evaluation of the statistical model. The present study could not provide well-established evidence for a relationship between shift work and hypercholesterolemia. PMID- 18427697 TI - Job control and the risk of incident stroke in the working population in Sweden. AB - OBJECTIVES: This study estimated the risk of incident stroke according to the level of job control and examined whether the association between job control and the risk of stroke varied as a function of gender. METHODS: This was a register based cohort study of nearly 3 million working people (age 30-64 years in 1990) with a 13-year follow-up (1991-2003) for incident stroke (50 114 events). Job control was aggregated to the data by a secondary data source (job-exposure matrix) in 1990. Gender-specific Cox regressions were applied. RESULTS: The age- and workhour-adjusted hazard ratio of the lowest versus the highest job control quartile was 1.25 [95% confidence interval (95% CI) 1.17-1.32] for any stroke, 1.33 (95% CI 1.15-1.55) for intracerebral hemorrhage, and 1.22 (95% CI 1.14-1.31) for brain infarction among the women, and the corresponding figures for the men were 1.24 (95% CI 1.21-1.28), 1.30 (95% CI 1.21-1.40), 1.23 (95% CI 1.19-1.28), respectively. Adjustment for education, marital status, and income attenuated these associations to 1.07 (95% CI 1.01-1.14) for any stroke, 1.22 (95% CI 1.04 1.42) for intracerebral hemorrhage, and 1.04 (95% CI 0.97-1.12) for brain infarction for the women and to 1.08 (95% CI 1.04-1.12), 1.12 (95% CI 1.03-1.22), 1.08 (95% CI 1.04-1.13), respectively, for the men. CONCLUSIONS: The relative risk of stroke was higher in low job-control occupations. The association between job control and stroke subtypes varied as a function of gender. The relative risk of intracerebral hemorrhage was highest for the women in low job-control occupations. PMID- 18427698 TI - Job strain and arteriosclerosis in three different types of arteries among male Japanese factory workers. AB - OBJECTIVES: This study evaluated the association between job strain and subclinical indicators of arteriosclerosis simultaneously in the cerebral artery, the aorta, and the carotid artery. METHODS: The participants in this cross sectional study consisted of 352 male factory workers between the ages of 24.9 to 55.8 (mean 41.7) years who had not been exposed to any hazardous chemicals in Japan. Job strain was measured using a Japanese version of the Job Content Questionnaire developed by Karasek. The following subclinical indicators of arteriosclerosis were examined by noninvasive procedures: hyperintense spots in T2-weighted images in brain magnetic resonance imaging (MRI), pulse wave velocity of the aorta, and the stiffness parameter of the carotid artery using ultrasound and systolic and diastolic blood pressure. RESULTS: After control for potential confounders, high job strain was associated with an increased prevalence of hyperintense spots (adjusted odds ratio 2.53, 95% confidence interval 1.14-5.63), and elevated pulse-wave velocity (adjusted increase in percent 3.5, 95% confidence interval 0.0-6.9). The stiffness parameter tended to be elevated in the high job-strain group although this trend was not statistically significant. Job strain showed no relation to blood pressure. CONCLUSIONS: In this study, job strain was associated with subclinical indicators of arteriosclerosis in different arteries although the association was not statistically significant for the carotid artery. New information is offered by brain MRI for arteriosclerosis in the cerebral artery. An increase in hyperintense spots may explain the link between job strain and cerebrovascular disease. PMID- 18427699 TI - One-year randomized controlled trial with different physical-activity programs to reduce musculoskeletal symptoms in the neck and shoulders among office workers. AB - OBJECTIVES: This study evaluates the effect of two different worksite physical activity interventions on neck-shoulder symptoms, together with perceived work ability and sick leave among office workers. METHODS: An examiner-blinded randomized controlled trial was conducted with 549 office workers allocated to one of three intervention groups: one with specific resistance training (SRT) of the neck-shoulder region (N=180), one with all-round physical exercise (APE) (N=187), and one which acted as a reference group, which was informed about general health-promoting activities but did not include a physical activity program (N=182). Questionnaires were filled out at baseline and after 1 year of training. RESULTS: The duration and intensity of neck and shoulder symptoms was lower after the specified worksite physical-activity interventions than in the reference group. On an intervention group level, SRT was not more effective than APE in reducing the duration and intensity of neck and shoulder symptoms. However, those asymptomatic at baseline had a significant lower prevalence of neck-shoulder symptoms at follow-up when allocated to the SRT group than placed in the APE group or reference group. At baseline the work ability index (WAI) was close to 90% of the maximum score, and the mean sick leave was 5 days per year, both being unaffected by the interventions. CONCLUSIONS: Different physical activity interventions were successful in reducing neck-shoulder symptoms, and SRT was superior to APE in the primary prevention of such symptoms. The initially relatively high WAI was the most probable reason for no further increase in WAI. Likewise the mean sick leave the year before the intervention was very low, and it was probably not possible to reduce it further. PMID- 18427700 TI - Asthma incidence in wood-processing industries in Finland in a register-based population study. AB - OBJECTIVES: This register-based population study determined incidence rates of clinically verified asthma among woodworkers, other blue-collar workers, and administrative personnel employed in wood-processing industries in Finland. Exposure to wood dust was under special scrutiny. METHODS: All Finns employed in wood-processing industries were followed for asthma incidence via record linkage in the years 1986-1998. Incident cases included people with asthma reimbursed for medication by the national health insurance or registered as having occupational asthma. Age-adjusted incidence rates and relative risks (RR) by gender were estimated for wood workers, other blue-collar workers, and administrative employees (referents) in wood industries. RESULTS: The relative risk of asthma was increased for all woodworkers among both genders [men: RR 1.5, 95% confidence interval (95% CI) 1.2-1.8; women: RR 1.5, 95% Cl 1.2-1.7]; a similarly elevated risk was also found for other blue-collar workers (men: RR 1.5, 95% Cl 1.2-1.8; women: RR 1.4, 95% Cl 1.2-1.6) in the same wood industries. Statistically increased relative risks were found for low and medium exposure to wood dust, but not for high exposure. Altogether 217 of the 4074 clinically verified asthma cases were reported as occupational asthma in the Finnish Register on Occupational Diseases. CONCLUSIONS: The incidence rates for asthma were significantly increased both among the woodworkers and the other blue-collar workers in wood industries but without a clear dose-response. Cases recognized as occupational asthma accounted for only a small part of the total asthma excess, indicating that much of the work-related asthma excess remains unrecognized in these industries. PMID- 18427701 TI - Cancer incidence among Finnish farmers, 1995-2005. AB - OBJECTIVES: The aim of this study was to describe the cancer risk pattern of male and female Finnish farmers and to define the role of the type of farm production as a risk determinant. METHODS: All farmers on 31 December 1978 living on 1 January 1995 were included in the cohort. The data concerning continuation as a farmer and the type of farming was collected from the 31 December 1990 and 1994 farm registries. The observed number of cases in each stratum was divided by the respective expected number based on national incidence rates to calculate the standardized incidence ratio. RESULTS: The overall cancer incidence was smaller than that of the general population. For most of the cancer sites, the standardized incidence ratios were below 1.0. The lowest rates for farmers continuing to farm were determined for mesothelioma and cancers of the liver, larynx, lung, nose, esophagus, and urinary bladder. The only significantly elevated standardized incidence ratio was that for lip cancer. Permanent beef and dairy farmers had the lowest standardized incidence ratios for overall cancer. Dairy farmers of 1978 who had changed their production type to crop farmers had increased their risk for overall cancer from a standardized incidence ratio of 0.82 to 0.92. CONCLUSIONS: The cancer incidence of Finnish farmers was significantly below the national average. The finding that lung cancer risk was low among dairy farmers but increased with a change to another type of farm production gives some support to the hypothesis that endotoxin exposure may decrease cancer risk. PMID- 18427702 TI - Deconstructing the 'black box' of the Camberwell assessment of need score in mental health services evaluation. AB - OBJECTIVE: The aim of the study was to examine an alternative way of scoring the Camberwell Assessment of Need (CAN) for the purpose of service evaluation, using the by us defined Social Services (SI) and Psychiatric Services (PI) subindices. METHODS: CAN assessments in 1997 and 1999 of 262 outpatients (mean age 45 years, 77.1% psychotic disorders) were reanalysed to fit the SI and the PI, which were compared to the full CAN. RESULTS: The mean total needs on the full CAN decreased from 6.65 to 6.22 (P = 0.007), as did the mean unmet needs (1.55-1.81, P = 0.049). The mean total needs on the PI decreased from 2.42 to 2.22 (P = 0.006), as did the mean unmet needs (1.66-0.57, P < 0.001). No changes in mean needs occurred on the SI. CONCLUSIONS: All significant changes occurred on the PI, indicating a more beneficial outcome of the psychiatric care than the social care in terms of meeting needs, a result impossible to discern from the total scores of the CAN. Thus, output scores on subindices of the CAN might be useful as outcome measures in service evaluation. PMID- 18427703 TI - Practice of electroconvulsive therapy at the research and training hospital in Turkey. AB - OBJECTIVE: We sought to obtain an overview of electroconvulsive therapy (ECT) practice in Bakirkoy Research and Training Hospital for Psychiatric and Neurological Diseases, which is the biggest hospital for psychiatry in Turkey. METHOD: From 1st January 2006 to 30th June 2007, a form enquiring about evaluation of ECT was filled retrospectively. RESULTS: The total number of patients, admitted for psychiatry during the survey period was 265,283. A total of 1,531 patients (12.4% among inpatients and 0.58% in all psychiatric admissions) received 13,618 sessions (including multiple hospitalizations) of ECT from 12,341 psychiatric inpatients during the survey period. Ninety-eight patients had multiple hospitalizations. The male-to-female ratio was 1.26-1. Patients with bipolar affective disorder, current episode manic with or without psychotic symptoms received ECT most frequently (30.3%), followed by patients with schizophrenia (29.5%), severe depressive episode with or without psychotic symptoms (include bipolar affective disorder current episode severe depression) (15.2%), other non-organic psychotic disorders (14.4%), schizoaffective disorders (6.3%), mental and behavioral disorders due to psychoactive substance abuse with psychotic disorders (3.5%) and catatonic schizophrenia (0.7%). Patients who received ECT were in age group of 25-44 years (64.7%), followed by 45-64 years (17.7%), 18-24 years (15.4%), 65 years and older (1.4%), and younger than 18 years (0.8%). All patients received modified ECT. There were no ECT-related deaths during the survey. CONCLUSION: The rate of ECT among all psychiatric inpatients during the survey period was 12.4%. The majority of patients who received ECT were diagnosed with bipolar affective disorder-current episode manic and schizophrenia. ECT training programs for psychiatry residents and specialists should be planned, and conducted systematically. PMID- 18427704 TI - User satisfaction with child and adolescent mental health services: impact of the service unit level. AB - BACKGROUND: Child and adolescent mental health service units (CAMHS) play an important role in the supply of services to children and adolescents with mental illness. The purpose of this study was to examine the service unit effect on parent satisfaction with outpatient treatment. METHOD: The study was undertaken in 49 of 72 Norwegian outpatient CAMHS in 2004. A total of 2253 of the parents who were asked to participate (87%) responded. Parent satisfaction was measured using two summated scales: clinician interaction/information and treatment outcome. Multilevel analyses were used to assess the contribution of the service units to satisfaction and to investigate patient level predictors of parent satisfaction. RESULTS: About 96-98% of the parent satisfaction variance could be attributed to factors within CAMHS, leaving only 2-4% of the variance attributable to the CAMHS level. Parents of patients aged 0-6 years were more satisfied than older patients' parents. Longer treatment episodes were positively associated with satisfaction. Parents whose children had been referred with externalizing symptoms were less satisfied with treatment outcome than those referred for internalizing symptoms. Waiting time was negatively associated with treatment outcome satisfaction. Adjustments for patient characteristics did not substantially change the relative effect of CAMHS on satisfaction ratings. CONCLUSION: The results indicate that information from user satisfaction surveys has clear limitations as an indicator of CAMHS quality. From a quality improvement perspective, the factors affecting the variance within CAMHS are of dominating importance compared to factors affecting between CAMHS variance. PMID- 18427705 TI - Public perceptions, explanatory models and service utilisation regarding mental illness and mental health care in Kenya. AB - The degree to which the mental health services are utilized depends partly on the public's views about mental illness and the public's perceptions about the roles of the providers of the services. Therefore, effective implementation of Kenya's mental health policy during the 1990s was likely to be influenced in some degree by how far it was compatible with the public's concepts of mental illness. The aim of this study was thus to explore the conceptual model underlying the views of the Kenyan public about mental illness and relate it to the national mental health policy of 1994. A household survey was conducted using a Key Informant Interview schedule. Results showed that the public subscribed to a biopsychosocial view of mental illness and health care, which was indeed compatible with the mental health policy, which was rooted in the concepts of Primary Health Care, as articulated at Alma Ata. However, the public did not expect biopsychosocial care from the health services, but rather only the biological/pharmacological component, relying on other care providers for psychosocial management. This discrepancy could have impeded the implementation of the 1994 policy. PMID- 18427706 TI - Uroflowmetry: its current clinical utility for women. AB - Uroflowmetry, the simple, non-invasive measurement of urine flow over time during micturition, has a long and interesting history, clear definitions, a clear purpose in screening for voiding difficulty and, most importantly, technical accuracy. Data interpretation is currently limiting its clinical utility, despite appropriate analysis being available in long-standing existing research. The main clinically important numerical parameters are the maximum and average urine flow rates and the voided volume. Urine flow rates are strongly dependent on voided volume. Reference to established (Liverpool) nomograms will most accurately correct for this dependency. Nomograms will also optimise the validation of uroflowmetry data and the accurate assessment of its normality, compared with fixed urine flow rates and "cutoffs" for voided volume. Abnormally slow urine flow (under the 10th centile Liverpool Nomograms) is the most clinically significant abnormality. Repeat uroflowmetry, concomitant post-void residual measurement and voiding cystometry studies are appropriate options for evaluating any abnormal uroflowmetry. PMID- 18427707 TI - Optimal decision criterion for detecting change in bone mineral density during serial monitoring: a Bayesian approach. AB - Interpretation of change in serial bone densitometry using least significant change (LSC) may not lead to optimal decision making. Using the principles of Bayesian statistics and decision sciences, we developed the Optimal Decision Criterion (ODC) which resulted in 11-12.5% higher rate of correct classification compared with the LSC method. INTRODUCTION: The interpretation of change in serial bone densitometry emphasizes using least significant change (LSC) to distinguish between true changes and measurement error. METHODS: Using the principles of Bayesian statistics and decision sciences, we developed the optimal decision criterion (ODC) based on maximizing a 'utility' function that rewards the correct and penalizes the incorrect classification of change. The relationship between LSC and ODC is demonstrated using a clinical sample from the Manitoba Bone Density Program. RESULTS: Under certain conditions, it can be shown that using LSC at the 95% confidence level implicitly equates the benefit of 39 true positive diagnoses with the harm of one false positive classification of BMD change. ODC resulted in an 11% higher rate of correct classification for lumbar spine BMD change and a 12.5% better performance for classifying total hip BMD change compared with LSC with this method. CONCLUSIONS: ODC has the same clinical interpretation as LSC but with two major advantages: it can incorporate prior knowledge of the likely values of the true change and it can be fine-tuned based on the relative value placed on the correct and incorrect classifications. Bayesian statistics and decision sciences could potentially increase the yield of a BMD monitoring program. PMID- 18427708 TI - Responses of planktonic microorganisms from tropical reservoirs to paraquat and deltamethrin exposure. AB - This study focused on the effects of two pesticides, paraquat (herbicide) and deltamethrin (insecticide), which are two common molecules used intensively in Burkina Faso. Natural bacterial populations, phytoplankton cultures (one cyanobacterium, Cylindrospermopsis raciborskii, and one chlorophycea, Monoraphidium sp.), and two species of zooplankton (Diaphanosoma excisum and Moina micrura) were isolated from aquatic communities and were used as biological targets in the experimental protocols. Paraquat was moderately toxic to bacteria and phytoplankton, whereas deltamethrin was significantly toxic only to the zooplankton species. Paradoxically, the chlorophycea Monoraphidium sp. exhibited a significant increase of in vitro fluorescence after 48 h at the highest doses. Preliminary tests were also performed from natural water extract of the main drinking water supply of the country (Loumbila Reservoir) by using solid-phase extraction. Obviously, the natural extract proved to be toxic to the same biological targets. Despite the absence of any determination of pesticides in the natural extract, the question of contamination and toxicity of these waters affects concerns about the safety of water supply and the effect of human pressure on the dynamics of planktonic communities of freshwater reservoirs in arid regions of western Africa. PMID- 18427709 TI - A Bayesian approach to probabilistic risk assessment in municipal playgrounds. AB - A Bayesian risk assessment of the exposure of children up to 6 years of age to trace elements in the sandy substrate of municipal playgrounds in Madrid, Spain was carried out. As a first step, exposure variables and parameters were borrowed from US Environmental Protection Agency (EPA) databases. However, the use of these non-site-specific exposure data introduced a high arbitrariness in the assumed distributions, which, in the Bayes approach, translates into a high uncertainty for those distributions that is also reflected in the risk outcome. In order to reduce this uncertainty, site-specific values for children's body weight and exposure frequency were determined from three surveys of 75, 56, and 34 individuals, respectively, carried out in randomly selected playgrounds in Madrid. This information was used in a Bayesian approach to modify the prior distributions of exposure frequency and body weight adapted from the literature. As a result, the predictive distributions of risk values for arsenic and mercury presented a reduced arbitrariness and a significantly lower uncertainty than those obtained either from the prior distributions borrowed from the US EPA or from the limited data gathered in Madrid separately. PMID- 18427710 TI - Nitrous oxide emissions from wetland rice-duck cultivation systems in Southern China. AB - Nitrous oxide (N2O) emissions from a rice-duck cultivation system in the subtropical region of China and its regulating factors were investigated by using a static chambers technique during rice growth seasons in 2006 and 2007. The experimental field was equally divided into six plots for two different treatments: One was a conventional rice field (CK) and the other was a rice-duck ecosystem (RD). With the same amount of urea applied as basal fertilization, N2O emission fluxes from RD and CK followed a similar seasonal variation trend. During the flooding seasons, the N2O emission flux was not correlated with temperature, but it was significantly related to soil inorganic nitrogen (SIN) (p < 0.01) and soil pH (p < 0.01). After drainage, the N2O emission flux was not correlated with temperature, SIN, and soil pH. Our experimental data showed that peaks of N2O emission flux occurred both in 2 weeks after urea application and after drainage. Compared to CK, RD could significantly increase N2O emission. We evaluated the integrated global warming potentials (GWPs) of a rice-duck cultivation system based on methane (CH4) and N2O emission, which showed that RD could suppress the total amount of CH4 and N2O emissions from rice paddies. Moreover, because the decrease of CH4 emissions from RD compared to CK was far more than the increase of N2O emissions from RD compared to CK, RD greatly reduced integrated GWPs (CH4 + N2O) compared to CK. So, the rice-duck cultivation system is an effective strategy for reducing integrated GWPs of the rice-duck cultivation systems based on CH4 and N2O in southern China and will contribute to alleviating global warming. PMID- 18427711 TI - An initial qualitative study of dual-energy CT in the knee ligaments. AB - OBJECTIVE: To study the clinical application of dual-energy CT (DECT) in the knee ligaments. METHODS: Twelve cases (24 knees) were scanned using dual-energy CT for the knee. Two- and three-dimensional images were used for display in all cases by means of multi-planar reformation (MPR) and volume rendering technique (VRT). All images were ranked by two radiologists according to the grade of knee ligament displayed, the definition of edge and attachment points of the knee ligament. RESULTS: The partial ligaments of 24 knees, such as the patellar ligament, fibular collateral ligament, anterior cruciate ligament (ACL) and posterior cruciate ligament (PCL) were clearly displayed; the tibial collateral ligament was not satisfactorily displayed. The transversal ligaments, such as lateral patellar retinaculum and medial patellar retinaculum, and the posterior ligament, such as oblique popliteal ligament could not be shown clearly. CONCLUSION: The dual-energy CT is a new and valuable tool to qualitatively display the main ligaments of the knee. PMID- 18427712 TI - Synthesis of bis-amino acid derivatives by Suzuki cross-coupling, Michael addition and substitution reactions. AB - Several bis-amino acids were prepared using a bis-Suzuki coupling (compounds 4-8, 10), a sequential Michael addition and bis-Suzuki coupling (compounds 12, 13) and a Michael addition followed by a substitution reaction (compounds 18, 19). Thus, the pure stereoisomer of the methyl esters of N-(tert-butoxycarbonyl)-beta bromodehydroaminobutyric acid and dehydrophenylalanine and of N-benzyloxycarbonyl beta-bromodehydroaminobutyric acid were reacted with 1,4-phenylene-bis-boronic acid or 9,9-dioctyl-9H-fluorene-2,7-bis-boronic acid using modified Suzuki coupling conditions. The corresponding bis-dehydroamino acid derivatives were obtained in good to high yields maintaining the stereochemistry of the starting materials. This reaction was also applied successfully to a brominated dehydrodipeptide and 1,4-phenylene-bis-boronic acid showing that it could be used to create cross-links in peptide chains. An N,N-diacyldehydroalanine derivative was used in a sequential Michael addition and bis-Suzuki coupling giving a p terphenyl bis-amino acid and a fluorenyl bis-amino acid in good yields. Two bis alpha,beta-diamino acids were obtained by a Michael addition of 1,2,4-triazole to the methyl esters of N-(4-toluenesulfonyl), N-(tert-butoxycarbonyl) dehydroamino acids followed by treatment with ethylenediamine. PMID- 18427713 TI - Using Chou's pseudo amino acid composition to predict protein quaternary structure: a sequence-segmented PseAAC approach. AB - In the protein universe, many proteins are composed of two or more polypeptide chains, generally referred to as subunits, which associate through noncovalent interactions and, occasionally, disulfide bonds to form protein quaternary structures. It has long been known that the functions of proteins are closely related to their quaternary structures; some examples include enzymes, hemoglobin, DNA polymerase, and ion channels. However, it is extremely labor expensive and even impossible to quickly determine the structures of hundreds of thousands of protein sequences solely from experiments. Since the number of protein sequences entering databanks is increasing rapidly, it is highly desirable to develop computational methods for classifying the quaternary structures of proteins from their primary sequences. Since the concept of Chou's pseudo amino acid composition (PseAAC) was introduced, a variety of approaches, such as residue conservation scores, von Neumann entropy, multiscale energy, autocorrelation function, moment descriptors, and cellular automata, have been utilized to formulate the PseAAC for predicting different attributes of proteins. Here, in a different approach, a sequence-segmented PseAAC is introduced to represent protein samples. Meanwhile, multiclass SVM classifier modules were adopted to classify protein quaternary structures. As a demonstration, the dataset constructed by Chou and Cai [(2003) Proteins 53:282-289] was adopted as a benchmark dataset. The overall jackknife success rates thus obtained were 88.2 89.1%, indicating that the new approach is quite promising for predicting protein quaternary structure. PMID- 18427714 TI - Prediction of protein structure class by coupling improved genetic algorithm and support vector machine. AB - Structural class characterizes the overall folding type of a protein or its domain. Most of the existing methods for determining the structural class of a protein are based on a group of features that only possesses a kind of discriminative information for the prediction of protein structure class. However, different types of discriminative information associated with primary sequence have been completely missed, which undoubtedly has reduced the success rate of prediction. We present a novel method for the prediction of protein structure class by coupling the improved genetic algorithm (GA) with the support vector machine (SVM). This improved GA was applied to the selection of an optimized feature subset and the optimization of SVM parameters. Jackknife tests on the working datasets indicated that the prediction accuracies for the different classes were in the range of 97.8-100% with an overall accuracy of 99.5%. The results indicate that the approach has a high potential to become a useful tool in bioinformatics. PMID- 18427715 TI - An ensemble of support vector machines for predicting the membrane protein type directly from the amino acid sequence. AB - Given a particular membrane protein, it is very important to know which membrane type it belongs to because this kind of information can provide clues for better understanding its function. In this work, we propose a system for predicting the membrane protein type directly from the amino acid sequence. The feature extraction step is based on an encoding technique that combines the physicochemical amino acid properties with the residue couple model. The residue couple model is a method inspired by Chou's quasi-sequence-order model that extracts the features by utilizing the sequence order effect indirectly. A set of support vector machines, each trained using a different physicochemical amino acid property combined with the residue couple model, are combined by vote rule. The success rate obtained by our system on a difficult dataset, where the sequences in a given membrane type have a low sequence identity to any other proteins of the same membrane type, are quite high, indicating that the proposed method, where the features are extracted directly from the amino acid sequence, is a feasible system for predicting the membrane protein type. PMID- 18427716 TI - Secondary structure-based assignment of the protein structural classes. AB - Structural class categorizes proteins based on the amount and arrangement of the constituent secondary structures. The knowledge of structural classes is applied in numerous important predictive tasks that address structural and functional features of proteins. We propose novel structural class assignment methods that use one-dimensional (1D) secondary structure as the input. The methods are designed based on a large set of low-identity sequences for which secondary structure is predicted from their sequence (PSSA(sc) model) or assigned based on their tertiary structure (SSA(sc)). The secondary structure is encoded using a comprehensive set of features describing count, content, and size of secondary structure segments, which are fed into a small decision tree that uses ten features to perform the assignment. The proposed models were compared against seven secondary structure-based and ten sequence-based structural class predictors. Using the 1D secondary structure, SSA(sc) and PSSA(sc) can assign proteins to the four main structural classes, while the existing secondary structure-based assignment methods can predict only three classes. Empirical evaluation shows that the proposed models are quite promising. Using the structure-based assignment performed in SCOP (structural classification of proteins) as the golden standard, the accuracy of SSA(sc) and PSSA(sc) equals 76 and 75%, respectively. We show that the use of the secondary structure predicted from the sequence as an input does not have a detrimental effect on the quality of structural class assignment when compared with using secondary structure derived from tertiary structure. Therefore, PSSA(sc) can be used to perform the automated assignment of structural classes based on the sequences. PMID- 18427717 TI - Protein function prediction with high-throughput data. AB - Protein function prediction is one of the main challenges in post-genomic era. The availability of large amounts of high-throughput data provides an alternative approach to handling this problem from the computational viewpoint. In this review, we provide a comprehensive description of the computational methods that are currently applicable to protein function prediction, especially from the perspective of machine learning. Machine learning techniques can generally be classified as supervised learning, semi-supervised learning and unsupervised learning. By classifying the existing computational methods for protein annotation into these three groups, we are able to present a comprehensive framework on protein annotation based on machine learning techniques. In addition to describing recently developed theoretical methodologies, we also cover representative databases and software tools that are widely utilized in the prediction of protein function. PMID- 18427718 TI - Prokaryotic diversity in one of the largest hypersaline coastal lagoons in the world. AB - Araruama Lagoon is an environment characterized by high salt concentrations. The low raining and high evaporation rates in this region favored the development of many salty ponds around the lagoon. In order to reveal the microbial composition of this system, we performed a 16S rRNA gene survey. Among archaea, most clones were related to uncultured environmental Euryarchaeota. In lagoon water, we found some clones related to Methanomicrobia and Methanothermococcus groups, while in the saline pond water members related to the genus Haloarcula were detected. Bacterial community was dominated by clones related to Gamma-proteobacteria, Actinobacteria, and Synechococcus in lagoon water, while Salinibacter ruber relatives dominated in saline pond. We also detected the presence of Alpha proteobacteria, Pseudomonas-like bacteria and Verrucomicrobia. Only representatives of the genus Ralstonia were cosmopolitan, being observed in both systems. The detection of a substantial number of clones related to uncultured archaea and bacteria suggest that the hypersaline waters of Araruama harbor a pool of novel prokaryotic phylotypes, distinct from those observed in other similar systems. We also observed clones related to halophilic genera of cyanobacteria that are specific for each habitat studied. Additionally, two bacterioplankton molecular markers with ecological relevance were analyzed, one is linked to nitrogen fixation (nifH) and the other is linked to carbon fixation by bacterial photosynthesis, the protochlorophyllide genes, revealing a specific genetic distribution in this ecosystem. This is the first study of the biogeography and community structure of microbial assemblages in Brazilian tropical hypersaline environments. This work is directed towards a better understanding of the free-living prokaryotic diversity adapted to life in hypersaline waters. PMID- 18427719 TI - Hyaluronan inhibits IL-1beta-stimulated collagenase production via down regulation of phosphorylated p38 in SW-1353 human chondrosarcoma cells. AB - We investigated the intracellular mechanism for the inhibitory effects of hyaluronan (HA) on interleukin-1beta (IL-1beta)-stimulated collagenase-1 and -3 (matrix metalloproteinases (MMPs)-1 and -13) production in a human chondrosarcoma cell line, SW-1353. MMPs-1 and -13 were induced by IL-1beta at 2 ng/ml in SW-1353 cells for 48 h. HA of 800 kDa, which is used clinically, significantly suppressed IL-1beta-stimulated production of MMPs-1 and -13 by immunoblotting. SW-1353 cells express the standard form of CD44 (CD44H), and immunofluorescent cytochemistry demonstrated the association of HA with CD44 on SW-1353 cells. Phosphorylated p38 (Phos-p38) mitogen-activated protein kinase was stimulated in SW-1353 cells by IL 1beta but not by HA alone. SB203580, a p38 MAPK inhibitor, partially blocked the MMP-1 and -13 production stimulated by IL-1beta. 800-kDa HA suppressed IL-1beta activated Phos-p38 in a dose-dependent manner. CD44 blocking significantly reversed the inhibitory effects of HA on IL-1beta-activated Phos-p38 production. The present study clearly suggests that HA binds CD44 and inhibits IL-1beta induced MMP-1 and -13 expression via down-regulation of Phos-p38 in SW-1353 cells. PMID- 18427720 TI - Decreased percentages of regulatory T cells in peripheral blood of patients with Behcet's disease before ocular attack: a possible predictive marker of ocular attack. AB - The current study investigates the role of regulatory T (Treg) cells in the pathogenesis of ocular attack in patients with Behcet's disease (BD). Nineteen BD patients with ocular complications (BDo), including 11 BD patients with ocular attack (BDa) and eight BD patients with inactive ocular complications (BDi), were studied. Four BD patients without ocular complications (BDwo) were also evaluated as controls. All patients were prospectively followed by our outpatient clinic between autumn 2004 and spring 2005. CD4+ CD25+bright T cells (Treg cells) from peripheral blood were measured by flow cytometry. The percentages of Treg cells in CD4+ T cells from BDo were significantly decreased before ocular attack compared with those after ocular attack. Moreover, surprisingly, these levels before attack were significantly lower than normal level, whereas the percentages of Treg cells in both BDi and BDwo patients were normal. Treg cells were significantly decreased in BDa before active ocular attack. These findings suggest that Treg cells play an important role in ocular attack in BD patients. In addition, decreased percentages of Treg cells may be a predictive marker of ocular attack in BD patients allowing treatment of BD patients before an ocular attack. PMID- 18427721 TI - Bilateral ureteral stenosis as a complication of Henoch-Schonlein vasculitis. PMID- 18427722 TI - Thrombotic thrombocytopenic purpura-hemolytic uremic syndrome and adult onset Still's disease: case report and review of the literature. AB - Thrombotic thrombocytopenic purpura-hemolytic uremic syndrome (TTP/HUS) is a multisystem disorder characterized by consumptive thrombocytopenia, microangiopathic hemolytic anemia, neurologic symptoms, renal function abnormalities, and fever. Coexistence of TTP/HUS and adult onset Still's disease (ASD) is extremely rare. We report the case of a 46-year-old woman who presented with fever, arthritis, myalgias, petechia on skin and confusion five years after the onset of ASD. Thrombocytopenia, renal failure, marked elevation lactate dehydrogenase, and red cell fragmentation on peripheral blood smear were observed. We made a diagnosis of TTP/HUS associated with ASD, according to physical examination and characteristic laboratory data. She recovered from the TTP/HUS following daily sessions of therapeutic plasma exchange with fresh frozen plasma replacement and glucocorticoid therapy. Awareness of the possible development of TTP/HUS in ASD is important for early diagnosis and treatment. PMID- 18427723 TI - Safety of long-term tacrolimus therapy for rheumatoid arthritis: an open-label, uncontrolled study in non-elderly patients. AB - In this study we focused on the safety of long-term tacrolimus therapy in non elderly patients with rheumatoid arthritis who were treated with tacrolimus or mizoribine in a previous double-blind study. The patients received oral tacrolimus at a dose < or =3 mg once daily for 76 weeks. The safety analysis population included 115 patients aged 20-64 years. Adverse drug reactions presented as symptomatic events in 39 patients (33.9%), laboratory abnormalities in 38 patients (33.0%), and infections in 19 patients (16.5%). The major reactions were gastrointestinal disorders and hypertension as symptomatic events, increases of creatinine, urinary N-acetyl-beta-D-glucosamidase and hemoglobin A1C as laboratory abnormalities, and the common cold syndrome as infections. After 76 weeks of tacrolimus treatment, the ACR20 response rates of patients who had also received tacrolimus during the preceding double-blind study was 61.5% (compared with the status at baseline in the preceding study). The corresponding response rate for patients who had previously received mizoribine was 66.0%. The mean blood concentration of tacrolimus was 3.8-4.8 ng/mL. In conclusion, safety profiles of tacrolimus treatment for long-term seems to be similar to those of previous studies in patients with rheumatoid arthritis. PMID- 18427724 TI - Effect of alendronate on glucocorticoid-induced osteoporosis in Japanese women with systemic autoimmune diseases: versus alfacalcidol. AB - Glucocorticoids-induced osteoporosis is a serious problem for patients with systemic autoimmune disease requiring relatively long-term glucocorticoid treatment. Effectiveness of alendronate for the prevention of glucocorticoids induced osteoporosis was evaluated in comparison with that of alfacalcidol in Japanese women with autoimmune disease excluding rheumatoid arthritis. Loss of bone mass was evaluated with bone mineral density (BMD) of lumber vertebrae, bone resorption was with urinary N-telopeptide for type I collagen (NTX), and bone formation was with serum bone-specific alkaline phosphatase (B-ALP). A total of 33 patients who were treated with oral glucocorticoids (>or=5 mg/day of prednisolone equivalence) for more than 6 months were randomized into two groups; alendronate group (n = 17) received 5 mg/day of alendronate, and alfacalcidol group (n = 16) received 1.0 mug/day of alfacalcidol for 24 months with glucocorticoids. The dose of alendronate was the maximal dose approved in Japan. BMD had tendency to decrease with alfacalcidol, while increase with alendronate. The difference in BMD change between the two groups was significant by 4.3% at 18 months and by 4.2% at 24 months (both P < 0.05). Bone resorption was significantly reduced only with alendronate; NTX was decreased by 28 to 35% at 6 to 24 months (P < 0.05), but not changed with alfacalcidol at 24 months. The bone formation was found to be unchanged according to the B-ALP measured between the two groups. In conclusion, the treatment of 5 mg alendronate daily is more effective than alfacalcidol for preventing the glucocorticoid-induced osteoporosis by the mechanism of reducing bone resorption in Japanese women with systemic autoimmune disease. PMID- 18427725 TI - Influence of system size and solvent flow on the distribution of wormlike micelles in a contraction-expansion geometry. AB - Viscoelastic wormlike micelles are formed by surfactants assembling into elongated cylindrical structures. These structures respond to flow by aligning, breaking and reforming. Their response to the complex flow fields encountered in porous media is particularly rich. Here we use a realistic mesoscopic Brownian Dynamics model to investigate the flow of a viscoelastic surfactant (VES) fluid through individual pores idealized as a step expansion-contraction of size around one micron. In a previous study, we assumed the flow field to be Newtonian. Here we extend the work to include the non-Newtonian flow field previously obtained by experiment. The size of the simulations is also increased so that the pore is much larger than the radius of gyration of the micelles. For the non-Newtonian flow field at the higher flow rates in relatively large pores, the density of the micelles becomes markedly non-uniform. In this case, we find that the density in the large, slowly moving entry corner regions is substantially increased. PMID- 18427726 TI - Interaction potential and near wall dynamics of spherical colloids in suspensions of rod-like fd-virus. AB - Averaged diffusivities of spherical colloids in solutions of rod-like particles, i.e. fd-virus, were measured with EWDLS and TIRM. While the experimentally observed near wall dynamics of the spheres in the absence of fd are well described by standard hydrodynamic theories, there are significant deviations at finite fd concentrations. Both experimental methods yield data which are significantly smaller than the theoretical predictions. PMID- 18427727 TI - Headaches related to triptans therapy in patients of migrainous vertigo. AB - Dizziness and vertigo are frequently reported by patients with migraine. In migrainous vertigo (MV), vertigo is causally related to migraine. Patients of MV usually have an attenuated or absent headache with their vertigo as compared with their usual headache of migraine. Here we report three female patients of MV in which administration of triptan was associated with induction (two patients) or exacerbation (one patient) of headache with disappearance of vertigo. We suggest that headache and vertigo of migraine may be inversely related to each other and suppression of one may induce or aggravate the other. PMID- 18427728 TI - Prevalence of migraine diagnosis using ID Migraine among university students in southern Turkey. AB - In this prospective study, we aimed to determine the prevalence of migraine and non-migraine headaches using a questionnaire, including ID Migraine, for university students. The study was designed cross-sectionally and a questionnaire consisting of 43 questions was applied to 1,256 students. The questionnaire consisted of questions related to demographic, social, curriculum, housing and headache characteristics of the subjects. Three-item screening questions of the ID Migraine test were included at the end of the questionnaire aimed at migraine diagnosis. The mean age of 1,256 students (529 females and 727 males) enrolled in this study was 21.9 +/- 2.1 years (17-31 years). Migraine-type headache was detected in 265 subjects (21.9%) based on the ID Migraine evaluation. Of these, 145 (54.7%) were female and 120 (45.3%) were male (female/male ratio: 1.2/1). Non migraine-type headache was identified in 864 subjects, with 357 females and 507 males. As a conclusion, ID Migraine screening test might be practical and beneficial when a face-to-face interview is not possible or to pre-determine the subjects for a face-to-face interview for migraine diagnosis in larger populations. PMID- 18427729 TI - The study of the feasibility of segmental bone defect repair with tissue- engineered bone membrane: a qualitative observation. AB - The objective of the study was to investigate the feasibility of intramembranous osteogenesis from tissue-engineered bone membrane in vivo. Bone marrow mesenchymal stem cells (MSCs) of rabbits were harvested, expanded and some of them were induced into osteoblasts. Porcine small intestinal submucosa (SIS) was converted by a series of physical and chemical procedures into a scaffold. MSCs and induced osteoblasts were seeded separately onto the scaffold, thus fabricating two kinds of tissue-engineered bone membrane. A total of 12 New Zealand rabbits were subjected to a surgical operation; a 15 mm bone segment, including the periosteum, was resected from the radius on both sides of each rabbit to create critical bone defects. The two kinds of tissue-engineered bone membrane and SIS (as control) were implanted randomly into the site of bone defect. The animals had radiographs and were killed after 4 weeks. The specimens were harvested and histological examination performed for evidence of osteogenesis. Bone tissue had formed in defects treated by the two kinds of tissue-engineered bone membrane at 4 weeks. This was supported by the X-ray and histological examination, which confirmed the segmental gap bridged by bone. There was no attempt to bridge in the bone defect treated by SIS. Tissue engineered bone membrane, constructed by seeding allogeneic cells on an xenogeneic and bio-derived scaffold, can repair critical bone defects successfully. PMID- 18427730 TI - The effects of raloxifene hydrochloride on ocular hemodynamics and visual function. AB - OBJECTIVE: To assess the comprehensive effects of raloxifene hydrochloride on retinal, choroidal and retrobulbar hemodynamics and on visual function in post menopausal women. DESIGN: Twenty-four post-menopausal women (age 55 +/- 3.8 years) were recruited for this cross-sectional study: 12 received placebo and 12 received raloxifene hydrochloride 60 mg once a day for 3 months. Baseline measurements of both groups included heart rate (HR), blood pressure (BP), visual acuity, contrast sensitivity and intraocular pressure (IOP) for both eyes. A comprehensive ocular blood flow (OBF) assessment was obtained for each patient in a randomly chosen study eye. Retinal blood flow data was obtained using confocal scanning laser Doppler flowmetry [Heidelberg Retinal Flowmeter (HRF)]. Color Doppler imaging (CDI) was used to assess retrobulbar hemodynamics in the ophthalmic, central retinal, short nasal and temporal posterior ciliary arteries. Baseline vision and hemodynamics in post-menopausal subjects were compared using paired Student's t tests, and the percentage change in baseline versus 3-month parameters was analyzed. RESULTS: There were no statistically significant differences between 3 months of raloxifene therapy and placebo in terms of age, HR, arterial or mean BP, visual acuity, contrast sensitivity, IOP or retinal or retrobulbar blood flow. CONCLUSION: Raloxifene therapy at 60 mg/day had no clinically significant impact on BP, IOP or OBF in post-menopausal women. PMID- 18427731 TI - Clinical biomarkers of kinase activity: examples from EGFR inhibition trials. AB - INTRODUCTION: Tumor response and duration of patient survival after treatment with inhibitors of the epidermal growth factor receptor (EGFR) varies considerably between different kinds of EGFR inhibitors, different combination schedules, but also between individual patients. DISCUSSION: Development and introduction of biomarkers into clinical practice is necessary to predict treatment response and thereby to individualize cancer therapy. Due to specific interactions of EGFR inhibitors with biological effects of irradiation, biomarkers are expected to differ for radiation oncology compared to application of the drugs alone or within chemotherapy treatment schedules and therefore need to be established and tested separately. OBJECTIVES: The review summarizes the current status of potential predictors for the effect of EGFR inhibitors used as single agents or in combination with chemotherapy. CONCLUSION: Based on this knowledge and on preclinical radiotherapy data, candidate biomarkers and further research strategies for radiation oncology are discussed. PMID- 18427732 TI - Practical approaches to proteomic biomarkers within prostate cancer radiotherapy trials. AB - INTRODUCTION: Proteomic biomarkers may be useful for monitoring therapeutic response and to triage cancer patients to best therapy. OBJECTIVES: In this review, we highlight the importance of specimen acquisition, preparation and analysis in radiotherapy proteomic studies. We also discuss practical approaches for the design and execution of clinical proteomic studies using our recent experience based on specimens accrued during prostate cancer radiation therapy. DISCUSSION AND CONCLUSIONS: Numerous proteomic methods are being employed, including high-throughput mass spectrometry and immunoassays, and using solid tissues, blood and urine for analysis. Given the potential complexity of cytokine and other protein responses, there is a need to assess proteomic signatures within serial samples as longitudinal studies during a course of fractionated radiotherapy (RT). PMID- 18427733 TI - Biomarkers of clinical trials using molecular inhibitors and radiotherapy: state of-the-art. Preface. PMID- 18427734 TI - Pancreatic cancer: from molecular pathogenesis to targeted therapy. AB - Pancreatic cancer is a deadly malignancy with still high mortality and poor survival despite the significant advances in understanding, diagnosis, and access to conventional and novel treatments. Though cytotoxic chemotherapy based on the purine analogue gemcitabine remains the standard approach in adjuvant and palliative setting the need for novel agents aiming at the main pathophysiological abnormalities and molecular pathways involved remains soaring. So far, evidence of clinical benefit, though small, exists only from the addition of the targeted agent erlotinib on the standard gemcitabine chemotherapy. Apart from the popular monoclonal antibodies and small molecules tyrosine kinase inhibitors, other novel compounds being tested in preclinical and clinical studies target mTOR, NF-kappaB, proteasome and histone deacetylase. These new drugs along with gene therapy and immunotherapy, which are also under clinical evaluation, may alter the unfavorable natural course of this disease. In this review we present the main pathophysiological alterations met in pancreatic cancer and the results of the florid preclinical and clinical research with regards to the targeted therapy associated to these abnormalities. PMID- 18427735 TI - Denitrification of highly alkaline nitrate waste using adapted sludge. AB - Uranium extraction and regeneration of ion exchange resin generates concentrated nitrate effluents (typically 500 to 10,000 ppm NO(3)-N) that are highly alkaline in nature (pH 9.0 to 11.0). It is difficult to remove nitrate from such solutions using standard physiochemical and biological methods. This paper reports denitrification of such wastes using preadapted sludge (biomass), which was acclimatized to different influent pH (7.5 to 11.5) in a sequencing batch reactor (4 l) for 2 months. Performance of the developed consortia was studied under different pH (7.5 to 12). Biomass denitrified the synthetic wastewater containing 1,694 ppm NO(3)-N at a pH of 10.5. Decrease in nitrite build up was observed at higher pH, which differs from the reported results. Kinetic analysis of the data showed that specific rate of nitrate reduction was highest (78 mg NO(3)-N/g MLSS/h) at higher pH (10.5). This was attributed to the acclimatization process. Thus, high-strength nitrate wastewater, which was highly alkaline, was successfully treated using preadapted sludge. PMID- 18427736 TI - Fed-batch production of gluconic acid by terpene-treated Aspergillus niger spores. AB - Aspergillus niger spores were used as catalyst in the bioconversion of glucose to gluconic acid. Spores produced by solid-state fermentation were treated with 15 different terpenes including monoterpenes and monoterpenoids to permeabilize and inhibit spore germination. It was found that spore membrane permeability is significantly increased by treatment with terpenoids when compared to monoterpenes. Best results were obtained with citral and isonovalal. Studies were carried out to optimize spores concentration (10(7)-10(10) spores/mL), terpene concentrations in the bioconversion medium and time of exposure (1-18 h) needed for permeabilization of spores. Fed-batch production of gluconate was done in a bioreactor with the best conditions [10(9) spores/mL of freeze-thawed spores treated with citral (3% v/v) for 5 h] followed by sequential additions of glucose powder and pH-regulated with a solution containing 2 mol/L of either NaOH or KOH. Bioconversion performance of the spore enzyme was compared with the commercial glucose oxidase at 50, 60, and 70 degrees C. Results showed that the spore enzyme was comparatively stable at 60 degrees C. It was also found that the spores could be reutilized for more than 14 cycles with almost similar reaction rate. Similar biocatalytic activity was rendered by spores even after its storage of 1 year at 20 degrees C. This study provided an experimental evidence of the significant catalytic role played by A. niger spore in bioconversion of glucose to gluconic acid with high yield and stability, giving protection to glucose oxidase. PMID- 18427737 TI - Selection and optimization of Bacillus atrophaeus inoculum medium and its effect on spore yield and thermal resistance. AB - Bacillus atrophaeus's spores are used as biological indicators to monitor sterilization processes and as a Bacillus anthracis surrogate in the development and validation of biosafety methods. The regular use of biological indicators to evaluate the efficiency of sterilization processes is a legal requirement for health services. However, its high cost hinders its widespread use. Aiming at developing a cost-effective inoculum medium, soybean molasses and nutrient supplemented vinasse were evaluated for their effectiveness in solid-state fermentation (SSF). In biomass production, the results demonstrated that all tested compositions favor growth by providing the nutritional demands of the microorganism. Optimum casein peptone and soybean molasses concentration (1.0%, 2.5%, or 4.0%) was determined by a 2((2-0)) factorial experimental design. The results have showed a positive influence of peptone on biomass production. In order to define peptone final concentration (4.0% or 6.0%), a 2(2) factorial experimental design was used. An optimized medium containing 4.0% soybean molasses and 4.0% casein peptone was similar in performance to a synthetic control medium (tryptone soy broth) in dry-heat thermal-resistant spore production by SSF. An experiment performed under optimum SSF conditions resulted in 1.9 x 10(10) CFU g(-1) dry matter with D (160 degrees C) = 5.2 +/- 0.2 min. PMID- 18427738 TI - Sequential production of amylolytic and lipolytic enzymes by bacterium strain isolated from petroleum contaminated soil. AB - Amylases and lipases are highly demanded industrial enzymes in various sectors such as food, pharmaceuticals, textiles, and detergents. Amylases are of ubiquitous occurrence and hold the maximum market share of enzyme sales. Lipases are the most versatile biocatalyst and bring about a range of bioconversion reactions such as hydrolysis, inter-esterification, esterification, alcoholysis, acidolysis, and aminolysis. The objective of this work was to study the feasibility for amylolitic and lipolytic production using a bacterium strain isolated from petroleum contaminated soil in the same submerged fermentation. This was a sequential process based on starch and vegetable oils feedstocks. Run were performed in batchwise using 2% starch supplemented with suitable nutrients and different vegetable oils as a lipase inducers. Fermentation conditions were pH 5.0; 30 degrees C, and stirred speed (200 rpm). Maxima activities for amyloglucosidase and lipase were, respectively, 0.18 and 1,150 U/ml. These results showed a promising methodology to obtain both enzymes using industrial waste resources containing vegetable oils. PMID- 18427739 TI - Production of cellulolytic enzymes by Aspergillus phoenicis in grape waste using response surface methodology. AB - The production of cellulolytic enzymes by the fungus Aspergillus phoenicis was investigated. Grape waste from the winemaking industry was chosen as the growth substrate among several agro-industrial byproducts. A 2 x 2 central composite design was performed, utilizing the amount of grape waste and peptone as independent variables. The fungus was cultivated in submerged fermentation at 30 degrees C and 120 rpm for 120 h, and the activities of total cellulases, endoglucanases, and beta-glucosidases were measured. Total cellulases were positively influenced by the linear increase of peptone concentration and decrease at axial concentrations of grape waste and peptone. Maximum activity of endoglucanase was observed by a linear increase of both grape waste and peptone concentrations. Concentrations of grape waste between 5 and 15 g/L had a positive effect on the production of beta-glucosidase; peptone had no significant effects. The optimum production of the three cellulolytic activities was observed at values near the central point. A. phoenicis has the potential for the production of cellulases utilizing grape waste as the growth substrate. PMID- 18427740 TI - BTX removal from polluted water through bioleaching processes. AB - In this study, benzene, toluene, and xylenes (BTX) removal from contaminated water by physical, chemical, and biological processes was studied. Results showed that air sparging in polluted water can reduce monoaromatic compounds from 140,000 to about 5 microg/l in only 1 h process with a gassing rate of 0.33 VVM. This method cannot be considered as a green technology as pollutants are only transferred from the liquid phase to the gas phase The ultimate objective of this research was thus to evaluate the efficiency of a strategy involving BTX adsorption by granular-activated charcoal (GAC) and subsequent regeneration of this support by a bioleaching process. Analysis of such processes requires the building of analytical tools able to accurately determine the contents of the contaminants in samples containing biomass to make possible the calculation of reliable material balances. Current investigation showed that BTX are readily trapped by GAC particles with low further release in the liquid medium whereas they remain at least partially available for in situ biodegradation. BTX adsorption onto the GAC was shown to reach maximum solute retention close to 350, 250, and 150 (as mg/g GAC) for xylenes, toluene, and benzene, respectively. This approach, which could afford efficient biological active carbon regeneration, is very promising for the removal of BTX compounds from water without any further environment damage. PMID- 18427741 TI - Biosurfactant production by Pseudomonas aeruginosa grown in residual soybean oil. AB - Pseudomonas aeruginosa PACL strain, isolated from oil-contaminated soil taken from a lagoon, was used to investigate the efficiency and magnitude of biosurfactant production, using different waste frying soybean oils, by submerged fermentation in stirred tank reactors of 6 and 10 l capacities. A complete factorial experimental design was used, with the goal of optimizing the aeration rate (0.5, 1.0, and 1.5 vvm) and agitation speed (300, 550, and 800 rpm). Aeration was identified as the primary variable affecting the process, with a maximum rhamnose concentration occurring at an aeration rate of 0.5 vvm. At optimum levels, a maximum rhamnose concentration of 3.3 g/l, an emulsification index of 100%, and a minimum surface tension of 26.0 dynes/cm were achieved. Under these conditions, the biosurfactant production derived from using a mixture of waste frying soybean oil (WFSO) as a carbon source was compared to production when non-used soybean oil (NUSO), or waste soybean oils used to fry specific foods, were used. NUSO produced the highest level of rhamnolipids, although the waste soybean oils also resulted in biosurfactant production of 75-90% of the maximum value. Under ideal conditions, the kinetic behavior and the modeling of the rhamnose production, nutrient consumption, and cellular growth were established. The resulting model predicted data points that corresponded well to the empirical information. PMID- 18427742 TI - Construction of protein chip to detect binding of Mitf protein (microphthalmia transcription factor) and E-box DNA. AB - A protein chip was constructed to detect the binding of microphthalmia-associated transcription factor (Mitf) and E-box DNA. Mitf, a key regulatory transcriptional factor of pigmentation-related genes such as tyrosinase, binds to specific sequence (CATGTG) in E-box DNA within the promoter of tyrosinase in the melanocytes. We produced Mitf as a maltose-binding protein (MBP) fusion protein in Escherichia coli, purified it using an affinity column, and immobilized it on beta-cyclodextrin-coated glass plate. Binding of Mitf to its target DNA, E-box oligomer, was monitored by surface plasmon resonance (SPR), SPR imaging (SPRi), and fluorescence-based system. Among these detection methods, fluorescence method was the most reliable. In this method, fluorescent intensity was proportional to the DNA concentration (up to 20 microM) and Mitf (up to 500 microg/ml). Kinetics of DNA binding with Mitf showed Langmuir isotherm, and its kinetic constants were determined. It is expected that Mitf-E-box DNA chip can be used as a screening tool for depigmenting agents in the cosmetic industry. PMID- 18427743 TI - Response surface optimization of the critical medium components for pullulan production by Aureobasidium pullulans FB-1. AB - Culture conditions for pullulan production by Aureobasidium pullulans were optimized using response surface methodology at shake flask level without pH control. In the present investigation, a five-level with five-factor central composite rotatable design of experiments was employed to optimize the levels of five factors significantly affecting the pullulan production, biomass production, and sugar utilization in submerged cultivation. The selected factors included concentration of sucrose, ammonium sulphate, yeast extract, dipotassium hydrogen phosphate, and sodium chloride. Using this methodology, the optimal values for concentration of sucrose, ammonium sulphate, yeast extract, dipotassium hydrogen phosphate, and sodium chloride were 5.31%, 0.11%, 0.07%, 0.05%, and 0.15% (w/v), respectively. This optimized medium has projected a theoretically production of pullulan of 4.44%, biomass yield of 1.03%, and sugar utilization of 97.12%. The multiple correlation coefficient 'R' was 0.9976, 0.9761 and 0.9919 for pullulan production, biomass production, and sugar utilization, respectively. The value of R being very close to one justifies an excellent correlation between the predicted and the experimental data. PMID- 18427744 TI - Cell-wall composition and accessibility to hydrolytic enzymes is differentially altered in divergently bred switchgrass (Panicum virgatum L.) genotypes. AB - The aims of this study were to understand the genotypic variability in cell-wall composition and cell-wall accessibility to enzymes in select switchgrass plants obtained from two different populations derived from a base population of octaploid cultivars. Population C+3 was developed by three breeding generations for high digestibility and population C-1 developed by one generation of breeding for low digestibility. Above-ground biomass from 12 selected genotypes, three each with high or low digestibility within each population, was analyzed for their cell-wall aromatics and polysaccharides. The ratio of p-coumaric acid/ferulic acid was greater (P < or = 0.05) for the high-lignin C-1 population over the low-lignin C+3 population, although the amounts of these two phenolics did not differ between populations. Combined values of guaiacyl + syringyl-lignin were consistently higher in genotypes from the C-1 population as compared to the genotypes from the C+3 population. Overall, p-coumaric acid was released by enzymes in greater amounts than ferulic acid in all these genotypes. Genotypes in the C-1 population exhibited lower dry weight loss as compared to the genotypes in the C+3 population after enzymatic digestion, suggesting changes in cell-wall architecture. Overall, our data highlight the phenotypic plasticity coded by the switchgrass genome and suggest that combining dry matter digestibility with other more specific cell-wall traits could result in genotypes with greater utility as bioenergy feedstocks. PMID- 18427745 TI - Corrective osteotomy after malunion of mid shaft fractures of the clavicle. AB - Displaced mid shaft fractures of the clavicle result in some degree of shortening and rotation. These fractures often heal with some degree of malunion which can be symptomatic. The question arises as to whether surgical correction of the deformity will relieve the symptoms associated with the malunion. Ten patients with a symptomatic malunion of the clavicle were treated by means of a corrective osteotomy with plate and screw fixation. Outcome measurement was a pre and postoperative DASH score, range of motion and patient satisfaction. At follow up after a mean duration of 37 months there was a significant improvement of the DASH score, eight patients were satisfied, and range of motion did not differ significantly. Two patients had a complication resulting from the surgical procedure. PMID- 18427746 TI - The tantalum screw for treating femoral head necrosis: rationale and results. AB - Femoral head necrosis (FHN) is a progressive pathology due to the failure of blood supply to the proximal femoral epiphysis, with consequent necrosis of the sub-chondral bone and collapse of the articular cartilage and loss of congruity between the head and the acetabulum. Borrowing the biological and mechanical principles from the vascularized fibular graft technique for the femoral neck, the tantalum screws have been introduced. They show an extraordinary porosity, osteoconductivity, biocompatibility and very good osteoinductivity. Vitreous tantalum can be processed to take the form of a screw, with a round medial extremity and a 25 mm threaded lateral extremity that can be inserted into the neck of the femur, thereby supporting the articular cartilage, stimulating the repair process, interrupting the interface between necrotic and healthy tissue and favoring local vascularization. We have drawn up a treatment protocol for early-stage FHN, based on the insertion of a tantalum screw into the femoral neck. The implant has a cylindrical shape, with a 10 mm diameter in the smooth part and 15 mm in the threaded part. It is available in different sizes from 70 to 130 mm, with 5 mm increments. The aim of the study is to describe the clinical and instrumental results of the tantalum screw for FHN. From June 2004 to June 2006 we performed 15 implants. The tantalum screw was inserted with an incision on the trochanteric region with traction and under X-ray control. For the clinical evaluation of the hip, we used the Harris hip score (HHS). For diagnosis and staging we used standard X-rays in two views and/or MRI, using the Steinberg classification (J Bone Joint Surg Br 77:34-41, 1995) and CT. In all cases, the osteonecrosis extended to not more than 30% of the joint surface and the cartilage was intact with no collapse. To assess the results, we compared the pre operative and the post-operative HHS, calculated the percentage differences between the two. We then compared X-rays, CT scans and MRI before and some time after the operation to assess whether the problem had been addressed, taking into account the intracancellous edema and the possible extension of necrosis. After an average follow-up period of 15.43 +/- 5.41 months, ten implants (seven patients out of ten) were examined and all but one patient showed a marked improvement in HHS (the average increase was 127.9%), with no further progression of the disease. We believe that this procedure can be suitable for young patients with limited first or second stage osteonecrosis. The objective for the foreseeable future is to resolve the pain, improve the quality of life and prevent or at least postpone arthroplasty. PMID- 18427747 TI - Peri-acetabular external fixation for hip disease: an anatomical study. AB - Hip distraction using cross joint articulated external fixation has been used by a number of orthopaedic centres for the treatment of osteoarthritis, chrondrolysis, and osteonecrosis, as an alternative to hip arthroplasty or arthrodesis in the adolescent and young adults . The hip, however, is problematic with respect to external fixation, as it lies deep, surrounded by powerful muscles and in intimate contact with major neurovascular structures and intrapelvic organs. The numbers of patients treated by this technique to date remains small and the technical detail and potential complications, with respect to the application of the external fixation, unclear. In this study we used anatomical information from CT scans combined with computer modelling of the hip and pelvis to identify safe screw positions in the periacetabular region. The surgical insertion of the pins was then performed on five cadavers and anatomical dissections undertaken to confirm the structures at risk. This study indicated that whilst there are a number of anatomic constraints, it is still possible to insert three pins with good divergence into the limited bony corridor of the peri acetabular region recognising that the more posterior of the laterally inserted pins poses a potential risk to the sciatic nerve if not inclined away from the sciatic notch. PMID- 18427748 TI - The dynamic condylar screw in the management of subtrochanteric fractures: does judicious use of biological fixation enhance overall results? AB - Subtrochanteric fractures are fraught with certain anatomic, biologic and biomechanical challenges. Evolution of implants like the Gamma nail, fixed-angle nail plates, compression hip screws and dynamic hip screws with trochanteric stabilization plates underlines a persistent quest for a better implant. We studied the dynamic condylar screw DCS as an implant on a series of 30 consecutive patients with subtrochanteric fractures. Our purpose was to assess this implant as a panacea for subtrochanteric fractures. All cases of AO type A and B were anatomically fixed, whereas type C was biologically plated. The idea was to assess the applicability and adaptability of the DCS. Fractures in 29 cases united, with one patient suffering from an implant failure. There were 17 excellent, 5 good, 5 fair and 3 poor results. The DCS is a definite advance over previous methods of treatment; when combined with the utilization of biological fixation techniques for comminuted fractures, can be relied upon to treat all types of subtrochanteric fractures. PMID- 18427749 TI - Safety and reliability of external fixation for basicervical and intertrochanteric fractures in high-risk elderly patients. AB - Forty elderly patients with basicervical and pertrochanteric fractures were managed with uniplanar AO external fixator under regional anaesthetic block of the femoral nerve and lateral cutaneous nerve of the thigh from April 2003 to March 2006. The mean age of the patients was 67.9 +/- 5.5 years. External fixator application was performed under radiological control after closed reduction had been obtained. Comorbid factors, duration of surgery, duration of hospitalisation, complications, walking ability, time to union and mortality rate were recorded. Patients were followed up for a mean period of 12 +/- 4.5 months. Superficial pin tract infection occurred in 13 patients, healing in varus >10 degrees and with shortening >2 cm occurred in six patients, and one patient suffered a spontaneous ipsilateral femoral neck fracture after removal of the fixator. The mean time for union was 10.4 +/- 1.2 weeks. Rapid union rate and minor complications obtained in the present study are comparable to those obtained with standard internal fixation techniques. Minimal intraoperative blood loss, short operative time and early patient mobilisation are advantages signifying uniplanar external fixator application under regional anaesthetic block to be a viable option in treatment of basicervical and pertrochanteric fractures in high-risk elderly patients. PMID- 18427750 TI - Monolateral external fixation for the progressive correction of neurological spastic knee flexion contracture in children. AB - The purpose of this study was to report the results of the surgical treatment of spastic knee flexion contracture using tenotomy and progressive correction by external fixator-distractor devices. The study design involved a prospective observational study of 16 knees in nine patients with spastic flexion contracture greater than 30 degrees . Treatment was indicated for both ambulatory and nonambulatory patients; and, in the latter group when sitting or personal hygiene was compromised. The average age was 11.6 years (range 10-17). Five of the patients were male and four female. There was one case of hemiplegia (11.1%), two cases of paraplegia (22.2%), and six cases of quadriplegia (66.7%). Six patients retained some walking capacity, while three had none. In all cases, distal lengthening of the hamstrings was carried out. A monolateral fixator with a gradual correction device was applied for a period of 4.8 weeks. The average follow-up was 26.6 months. The preoperative straight-leg raise was 55 degrees . The popliteal angle was 58 degrees preoperatively (range 30-80 degrees ), 8.5 degrees on removal of the fixator, and 20 degrees at the end of the follow-up. COMPLICATIONS: There were no superficial or deep infections, and no fractures or distal sensory-motor alterations. There was one case of arthrodiatasis of the knee (6.3%) which was resolved when the fixator was removed, and 11 cases of pin track infection (68.7%) which were resolved with local care and oral antibiotics. To conclude, spastic knee flexion contracture can be treated gradually with monolateral external fixator with distraction devices, and with distraction modules which prevent acute stretching of the posterior neurovascular structures of the knee. PMID- 18427751 TI - Limb reconstruction surgery in China: an evaluation of its role in the largest developing nation. AB - China is the "economic tiger" of the twenty-first century. Its new economic policies have overturned the outlook and futures of many industries in this nation, and parallel to this industrial progress have been strides in medicine. However a country steeped in a history of several millennia does not replace generations of experience nor knowledge in a few decades. The author explores the history of limb reconstruction surgery in this country, setting it in context of a population exposed to both traditional Chinese medicine and Western medicine. PMID- 18427752 TI - Safe corridors in external fixation: the lower leg (tibia, fibula, hindfoot and forefoot). AB - It is a prerequisite for surgeons using external fixation systems to be familiar with safe corridors for half pin or wire insertion. Several atlases of cross sectional anatomy are available, mostly in print, to provide guidance [1-3]. This series of articles provides high-quality cross-sectional anatomical images together with guidance in locating safe corridors; the added benefit here is that the information can be downloaded and stored on the surgeon's or in the operating room computer for ease of reference. This review covers the lower leg from knee joint to foot. PMID- 18427753 TI - Quality of training in radiation oncology in Germany. Results of a 2006 survey. AB - PURPOSE: To evaluate residents' satisfaction with their training in radiation oncology, the first nationwide survey was done in 2006. Results were presented at the 2006 annual meeting of the German Society of Radiation Oncology (DEGRO). MATERIAL AND METHODS: A questionnaire with 39 questions regarding training in radiation oncology in Germany was developed and sent by e-mail. Questionnaires were returned by mail and analyzed anonymously. RESULTS: 96 questionnaires were received. A total of 88% of respondents are pleased with their decision of training in radiation oncology. Residents are strongly motivated by their interest in oncology. Quality of training is heterogeneous and not optimal. Training in three-dimensional treatment planning, radiochemotherapy and intracavitary brachytherapy is judged adequate, whereas special techniques such as intensity-modulated radiotherapy (IMRT) and permanent prostate implants are not covered by the majority of institutions. Organization of training in the departments is often judged insufficient. CONCLUSION: Radiation oncology is attractive for young doctors. However, training quality for radiation oncologists in Germany was judged to be heterogeneous and needs to be optimized. For this, results of this survey may be helpful. The overall positive judgment may help to attract more students into the field of radiation oncology, an issue that becomes increasingly important given the shortage of doctors and the strong competition with other disciplines. Modern techniques, such as IMRT, need to be integrated into training programs in order to maintain the high standard of radiation oncology in Germany. PMID- 18427754 TI - [Radiotherapy for shoulder impingement]. AB - BACKGROUND AND PURPOSE: Up to now, degenerative shoulder diseases were summarized by the term "periarthritis humeroscapularis". Actual shoulder diseases can be differentiated etiopathologically according to a primary and secondary impingement syndrome. Narrowing of the subacromial space, which is caused by an osseous shape variant, leads to primary impingement. Secondary impingement develops, when the subacromial space is reduced by swelling tissue below the osseous shoulder roof. This study aimed for the exact diagnosis to indicate therapy and to classify the results according to the Constant score. PATIENTS AND METHODS: From August 1999 to September 2002, 102 patients with 115 shoulder joint conditions underwent radiation therapy (RT). All joints received two RT series (6 x 0.5 Gy/series) applied in two to three weekly fractions, totaling a dosage of 6.0 Gy (250 kV, 15 mAs, 1-mm Cu filter). The second RT course started 6 weeks after the end of the first. 115 shoulders were examined before RT, 6 weeks after the second RT course and, finally, during the follow-up from January to May 2003. RESULTS: Pain relief was achieved in 94/115 shoulder joints (82%) after 18-month follow-up (median). A significant difference existed between secondary impingement and primary/non-impingement according to response. Tendinosis calcarea, bursitis subdeltoidea, tendovaginitis of the long biceps tendon, and capsulitis adhaesiva responded well to therapy. CONCLUSION: Shoulder diseases of secondary impingement demonstrate a good response to RT. Less or no benefit was found in primary impingement syndrome or complete rotator cuff disruption and acute shoulder injuries, respectively. PMID- 18427755 TI - A new scoring system to predicting the survival of patients treated with whole brain radiotherapy for brain metastases. AB - PURPOSE: To create a scoring system to estimate survival of patients who received whole-brain radiotherapy (WBRT) for brain metastases. MATERIAL AND METHODS: Based on a multivariate analysis of 1,085 retrospectively analyzed patients, a scoring system was developed. This score was based on the four significant prognostic factors found in the multivariate analysis including: age, performance status, extracranial metastases at the time of WBRT, and interval between tumor diagnosis and WBRT. The score for each prognostic factor was determined by dividing the 6 month survival rate (in %) by 10. The total score represented the sum of the partial scores for each prognostic factor. Total scores ranged from 9 to 18 points, and patients were divided into four groups. For each group, survival was compared for short-course (5 x 4 Gy) versus longer-course WBRT (10 x 3 Gy/20 x 2 Gy). RESULTS: Actuarial 6-month survival rates were 6% for patients with scores of 9-10 points, 15% for those with scores of 11-13 points, 43% for those with scores of 14-16 points, and 76% for those with scores of 17-18 points (p < 0.001). Longer-course WBRT was not associated with better survival than short course WBRT in any of the four groups. CONCLUSION: Patients with brain metastases receiving WBRT can be grouped with this score to estimate survival. Short-course and longer-course WBRT resulted in similar survival in all groups studied. However, in the more favorable patients with scores of 17-18, longer-course WBRT with lower doses per fraction should be considered, as these schedules have been associated with less neurocognitive toxicity. PMID- 18427756 TI - Radiotherapy of liver metastases. Comparison of target volumes and dose-volume histograms employing CT- or MRI-based treatment planning. AB - PURPOSE: To assess differences in delineated target volumes of liver metastases using contrast-enhanced CT and different MRI sequences for radiation treatment planning. PATIENTS AND METHODS: 25 patients with 43 colorectal liver metastases were recruited. Tumor margins were defined by two experienced radiologists. The resulting D90 was assessed and the CT-based 3-D dose distribution merged with the according MRI dataset by employing image fusion. A theoretical D90 as a result of MRI-based treatment planning was assessed for various MRI sequences individually. RESULTS: In venous phase contrast-enhanced CT, the mean tumor volume was 20 ml; T1-weighted (T1w) MRI, 27 ml; contrast-enhanced T1w 42 ml; T2w 65 ml. The difference between the target volumes as assessed by either CT or MRI was 181% for T1w images, 178% for contrast-enhanced T1w, and 246% for T2w sequences. All differences were statistically significant (p < 0.05). The analysis of the dose volume histograms revealed statistically significant differences (i.e., for the D90) for the different target volumes specified by CT and MRI: mean D90 on CT, 18 Gy; plain T1w, 16 Gy; contrast-enhanced T1w, 15.5 Gy; T2w, 12 Gy. Hence, delineation of a larger target volume in T2w MRI compared to contrast-enhanced CT resulted in a smaller D90. The mean differences of tumor volumes assessed by CT and plain T1w were significantly higher in the group of patients showing local tumor recurrences as compared to patients with long-term local tumor control (p = 0.002). CONCLUSION: For treatment planning of liver metastases, the use of either plain T1w or T2w sequences is recommended to delineate the clinical target volume as completely as possible and not to miss potential tumor cell congregations in the surroundings as in CT. PMID- 18427758 TI - Similar extent of apoptosis induction at doses of X-rays and neutrons isoeffective for cell inactivation. AB - BACKGROUND AND PURPOSE: The relative biological effectiveness (RBE) of neutrons differs for various biological endpoints, and for various cell and tissue types. With respect to the apoptosis induction, a whole range of values can be found in the literature, but the decisive factors are not clear. Most previous studies have used apoptosis-prone hematopoietic cells, whereas tumor cells have received little attention. The authors therefore decided to investigate apoptosis induction caused by X-rays and neutrons in a line of human melanoma cells, at doses which are isoeffective for the loss of colony-forming ability. MATERIAL AND METHODS: Human melanoma cells Be11, expressing p53 wild-type protein, were used throughout. Exponentially growing cells were exposed to two pairs of isoeffective doses (at surviving levels 10% and 1%) of 240-kV X-rays and 5.8-MeV neutrons. 1-8 days after irradiation, the frequency of apoptosis in adherent cells was assessed by two-parameter flow cytometric analysis with a DNA-dye-exclusion annexin-V binding assay as well as by morphological examination with DAPI staining. RESULTS: Apoptosis was induced most significantly 6-7 days after irradiation. The time courses, as well as the magnitudes of apoptosis induction, after isoeffective doses of X-rays and neutrons with respect to loss of colony-forming ability appeared to be comparable. RBE values in the range of 4-5 were estimated for apoptosis 4-8 days after irradiation by both the annexin V assay and morphological examination. CONCLUSION: Radiation-induced apoptosis depends on ionization density in the same way as cell inactivation in general does, i.e., the RBE is similar, and the ratio of cells dying by apoptosis to cells dying otherwise does not depend on radiation quality. PMID- 18427757 TI - Multisegmented tangential breast fields: a rational way to treat breast cancer. AB - PURPOSE: Using three-dimensional conformal radiation therapy (3D-CRT) and multisegmented conformal radiation therapy (MS-CRT) for breast cancer treatment, the dose coverage of the planning target volume (PTV) and the radiation burden on the organs at risk (OARs) were evaluated. MATERIAL AND METHODS: 3D-CRT and MS-CRT were planned for 436 unilateral breasts (217 left). All patients were treated with MS-CRT between 2005 and 2007. For PTV delineation and beam orientation, supportive structures were applied. The mean PTV was 1,130 cm3 (in ten patients > 2,200 cm3). Three-dimensional planning with weight-optimized medial and lateral open fields at a total dose of 50.4/1.8 Gy was followed by multisegmented planning with a reasonably high-dose-level dose cloud to define the medial subfield, and renewed optimization. This was repeated for the lateral subfield with a final optimization. For PTV coverage evaluation, the ICRU 50 was considered: the PTV portions receiving 95-107%, < 95% and > 107% of the prescribed dose (PTVD95- 107%, PTVD107%), and the PTV maximal dose (PTVDmax). To compare the OAR radiation burdens, the mean doses to the ipsi /contralateral lung, contralateral breast, and whole heart were documented. RESULTS: The multisegmented plans furnished significantly (p < 0.0001) better target coverage (PTVD95-107% 82.8% vs. 90.9%, PTVD107% 5.9% vs. 0.3% and PTVDmax 56.6 vs. 54.3 Gy). The mean OAR doses remained almost unchanged: ipsilateral lung 10.5 versus 10.4 Gy, contralateral lung 0.4 versus 0.4 Gy, contralateral breast 0.8 versus 0.8 Gy, and whole heart (for left-sided cancers) 4.8 versus 4.8 Gy. The subfields required a mean of 9.8 MU (monitor units), i.e., a mean total 7.6 MU increment. The planning took 10-20 min, and the delivery 5-10 min. CONCLUSION: MS-CRT is a good alternative to breast intensity modulated radiation therapy (IMRT) and seems adequate for right-sided cancers, whereas left-sided cancers necessitate a longer follow-up of heart-related side effects before a final assessment. PMID- 18427759 TI - Intracerebral cavernous hemangioma after cranial irradiation in childhood. Incidence and risk factors. AB - BACKGROUND AND PURPOSE: Radiotherapy is an integral part of various therapeutic regimens in pediatric and adult oncology. Endocrine dysfunction, neurologic and psychiatric deficits, secondary malignancies and radiation-induced necrosis are well-known possible late effects of cranial irradiation. However, only sporadic cases of radiation-induced cavernous hemangiomas (RICH) have been reported so far. PATIENTS AND METHODS: Pediatric patients who underwent cranial radiation therapy for malignant diseases between January 1980 and December 2003 were retrospectively analyzed. After the end of therapy they entered a detailed follow up program. RESULTS: Of 171 patients, eight (three patients with medulloblastoma, three patients with acute lymphoblastic leukemia, and one patient each with ependymoma and craniopharyngioma) developed intracerebral cavernoma 2.9-18.4 years after irradiation representing a cumulative incidence (according to the Kaplan-Meier method) of 2.24%, 3.86%, 4.95%, and 6.74% within 5, 10, 15, and 20 years following radiation therapy, respectively. In patients treated in the first 10 years of life, RICH occurred with shorter latency and significantly more often (p = 0.044) resulting in an even higher cumulative incidence. CONCLUSION: These findings and previously published cases show that cavernous hemangiomas may occur after irradiation of the brain several years after the end of therapy irrespective of the radiation dose and type of malignancy. Particularly children < 10 years of age at the time of irradiation are at higher risk. Since patients with RICH frequently do not show symptoms but hemorrhage is a possible severe complication, imaging of the central nervous system should be performed routinely for longer follow- ups, particularly in patients who were treated as young children. PMID- 18427760 TI - [Statin therapy for systolic heart failure. The CORONA trial (Controlled Rosuvastatin Multinational Trial in Heart Failure)]. PMID- 18427761 TI - [A 42 year old patient with bilateral loss of sight and hypertension. Gemcitabine associated thrombotic microangiopathy (TMA)]. AB - We report a case of a 42 year old male patient with a history of adrenocortical carcinoma, who was admitted with bilateral loss of sight and hypertension. Laboratory tests and further clinical evaluation showed hemolytic anemia, thrombocytopenia and acute renal failure. This was consistent with thrombotic microangiopathy / hemolytic uremic syndrome (HUS) due to gemcitabine therapy. The patient was successfully treated with prednisolon and antihypertensive drugs. Visus was completely restored, plasmapheresis was not needed. Clinicians should be aware of HUS as a rare complication of gemcitabine therapy. PMID- 18427762 TI - Yellowjackets (Vespula pensylvanica) thermoregulate in response to changes in protein concentration. AB - Social insects can modulate body temperature to increase foraging efficiency; however, little is known about how the relative value of protein resources affects forager body temperature. Such regulation may be important given that colony growth is often limited by protein availability. In this paper, we present what are, to our knowledge, the first data for social insects showing that thoracic temperatures (T (th)) of foragers increase with the protein content of food resources. In an introduced population of western yellowjacket (Vespula pensylvanica), we measured T (th) of foragers collecting high-quality protein (100% canned chicken) and low-quality protein (50% canned chicken, 50% indigestible alpha-cellulose by volume) at different ambient air temperatures (T (a)). Wasps foraging on 100% chicken consistently exhibited higher T (th) compared to wasps foraging on 50% chicken. After correcting for T (a), the mean T (th) for wasps collecting 100% chicken were 1.98 degrees C higher than those of individuals collecting 50% chicken. We suggest that this mechanism may increase foraging efficiency in this and other social wasp species. PMID- 18427763 TI - Plant origin of Okinawan propolis: honeybee behavior observation and phytochemical analysis. AB - Propolis is a natural resinous product collected by honeybees from certain plants. It has gained popularity as a food and alternative medicine. Poplar and Baccharis are well known as the source plants of European and Brazilian propolis, respectively. However, the propolis from Okinawa, Japan, contains some prenylflavonoids not seen in other regions such as Europe and Brazil, suggesting that the plant origin of Okinawan propolis is a particular plant that grows in Okinawa. To identify the plant origin of Okinawan propolis, we observed the behavior of honeybees as they collected material from plants and caulked it inside the hive. Honeybees scraped resinous material from the surface of plant fruits of Macaranga tanarius and brought it back to their hive to use it as propolis. We collected samples of the plant and propolis, and compared their constituents by high-performance liquid chromatography with a photo-diode array detector. We also compared their 1,1-diphenyl-2-picryl-hydrazyl radical scavenging activity. The chemical constituents and biological activity of the ethanol extracts of the plant did not differ from those of propolis. This indicates directly that the plant origin of Okinawan propolis is M. tanarius. PMID- 18427764 TI - [Wish vs. will]. AB - With unclear use of the words 'wish' and 'will', a differentiation can be sought by mean of the structural dynamic conception. There exists a clear difference between undecided projection of autopractic wish and goal-oriented behaviour guided by the intentional will. Prerequisites of the human will are a developed mental structure and its system of values. The various degrees of autonomy, outside of neurobiologic factors, guarantee the person's relative compatibilistic freedom. PMID- 18427765 TI - [End-of-life decisions, powers of attorney, and advance directives]. AB - In modern medicine, decisions about the kind of treatment at life's end are often inevitable. According to German law, powers of attorney and advance directives can be of help in these decisions. When a patient in a state of competence has issued a lasting power of attorney, there is no need for courts to appoint a proxy, and physicians immediately have a legally empowered decision-maker they can address. According to current German law, advance directives are legally valid and binding expressions of a patient's will. They are, however, more powerful when issued after consultation with a physician. If treatment at life's end no longer complies with the patient's will or loses its medical indication, the goal of treatment should be redirected towards palliation. This implies that life-sustaining treatment may be withdrawn or withheld, which is best accomplished with sensitivity to the needs of patients, relatives, and health care professionals. PMID- 18427766 TI - [Histopathology of urothelial carcinomas: crucial for patient management]. AB - Urothelial carcinomas include tumors with very different clinical outcomes: 70 80% of urothelial carcinomas are genetically stable and are associated with a favorable prognosis, but 20-30% are genetically unstable and have a high progression rate. Therefore, the current World Health Organization classification (2004) differentiates the histologic grade as low grade (LG) or high grade (HG). Unequivocal language is mandatory to optimize therapy and assess prognosis. In addition to TNM classification and histologic grade, genetic factors such as mutations of p53, fibroblast growth factor receptor 3 (FGFR3), and phosphatidylinositol-3-kinase (PIK(3)CA) are relevant in patients' prognoses. PMID- 18427767 TI - [Technical innovations in endourological stone therapy]. AB - Endoscopic stone management is a field of consistent technical innovation. For ureterorenoscopy, flexible scopes capable of 270 degrees deflection have allowed access to the complete urinary tract, including the lower-pole renal calculi. To counter the flaw of high repair costs, new-generation flexible ureterorenoscopes feature significantly greater durability and a less frequent need for repair. The development and improvement of auxiliary instruments such as access sheaths has facilitated the procedures, reduced complications, and improved the stone-free rates. A similar trend is seen for percutaneous nephrolitholapaxy (PCNL). Development is directed at miniaturised instruments such as the mini-PCNL. Especially in combination with a tubeless procedure in which a routine nephrostomy after PCNL is omitted and the nephrostomy tract is closed using hemostyptic sealant, similar success rates are achieved, with fewer complications and shorter hospital stays. PMID- 18427768 TI - [Transrectal sonography biofeedback training (TSB). Incontinence training after radical prostatectomy]. AB - BACKGROUND: Biofeedback methods are used in severe incontinence after radical prostatectomy (RP). Although contraction of the urethral sphincter can be demonstrated very well to the patient by video urethrocystoscopy, this method is invasive. Transrectal sonography biofeedback training (TSB) is a less invasive method for visualization of contractions of the musculus sphincter urethrae externus. PATIENTS AND METHODS: A total of 100 patients with urinary incontinence after RP either lost 50 g urine or more during a 1-h pad test or they could not feel their sphincter contraction during physiotherapy continence training. Patients lying on their left side followed the examination with a 7.5-MHz ultrasound probe on a second monitor. Patients were then asked to reduce the urinary stream for 5 s with 10%, 30%, 70% and 100% of maximum possible strength. A reduction in the diameter of the fluid column just above the sphincter urethrae externus proportional to the strength of contraction could be demonstrated to the patients. RESULTS: Pain prevented TSB in 8%; 8% reported on severe, 5% on moderate and 24% on mild pain. In 4% the sphincter could not be precisely detected. A total of 88% showed differentiated tension, while 96% claimed that TSB was helpful for restoring continence after radical prostatectomy. CONCLUSION: TSB, particularly in the transversal plane, is a viable method to treat urinary incontinence after radical prostatectomy. In 8% of patients pain prevented TSB, in 4% TSB did not lead to evaluable results, while in 88% TSB demonstrated the effects of different tension strength of the sphincter muscle to the patients. The method is very well accepted by the patients themselves. PMID- 18427769 TI - Chlorophyll breakdown during pepper fruit ripening in the chlorophyll retainer mutation is impaired at the homolog of the senescence-inducible stay-green gene. AB - The pepper chlorophyll retainer (cl) mutation is characterized by inhibition of chlorophyll degradation during fruit ripening. Ripe fruit of cl pepper containing chlorophyll and red carotenoids is brown, while ripe fruit containing chlorophyll and yellow carotenoids is green. In addition to the inhibitory effect during fruit ripening caused by cl, we show that chlorophyll degradation is inhibited during natural and dark-induced leaf senescence. Therefore, the cl mutation has the characteristics of the stay-green (sgr) mutants described in many other species. Upon the recent discovery of the SGR gene in various plant species, we isolated pepper SGR (CaSGR) and found that it genetically cosegregates with cl in a BC1 mapping population. Furthermore, sequencing the wild-type and mutant alleles revealed an amino-acid substitution of tryptophan (aromatic amino acid) to arginine (basic amino acid) at position 114 in the protein sequence. The single-nucleotide polymorphism (SNP) that differentiates the wild-type and mutant alleles was exploited to develop a PCR marker useful for marker-assisted selection. Expression of CaSGR as measured by semiquantitative RT-PCR was mostly induced upon fruit ripening and to a lesser extent upon leaf senescence. Taking together, our genetic, sequence and expression data all indicate that CaSGR is a candidate for controlling the cl mutation in pepper. PMID- 18427770 TI - Identification of quantitative trait loci controlling partial clubroot resistance in new mapping populations of Arabidopsis thaliana. AB - To date, mechanisms of partial quantitative resistance, under polygenic control, remain poorly understood, studies of the molecular basis of disease resistance have mainly focused on qualitative variation under oligogenic control. However, oligogenic conferred resistance is rapidly overcome by the pathogen and knowledge of the relationship between qualitative and quantitative resistance is necessary to develop durably resistant cultivars. In this study, we exploited the Arabidopsis thaliana-Plasmodiophora brassicae pathosystem to decipher the genetic architecture determining partial resistance. This soil-borne pathogen causes clubroot, one of the economically most important diseases of Brassica crops in the world. A quantitative trait locus (QTL) approach was carried out using two segregating populations (F(2) and recombinant inbred lines) from crosses between the partially resistant accession Burren and the susceptible accession Columbia. Four additive QTLs (one moderate and three minor) controlling partial resistance to clubroot were identified, all the resistance alleles being derived from the partially resistant parent. In addition, four epistatic regions, which have no additive effect on resistance, were also found to be involved in partial resistance. An examination of candidate genes suggested that a potentially diverse array of mechanisms is related to the different QTLs. By fine-mapping and cloning these regions, the mechanisms involved in partial resistance will be identified. PMID- 18427771 TI - Genetic analysis of opaque2 modifier loci in quality protein maize. AB - Quality protein maize (QPM) was created by selecting genetic modifiers that convert the starchy endosperm of an opaque2 (o2) mutant to a hard, vitreous phenotype. Genetic analysis has shown that there are multiple, unlinked o2 modifiers (Opm), but their identity and mode of action are unknown. Using two independently developed QPM lines, we mapped several major Opm QTLs to chromosomes 1, 7 and 9. A microarray hybridization performed with RNA obtained from true breeding o2 progeny with vitreous and opaque kernel phenotypes identified a small group of differentially expressed genes, some of which map at or near the Opm QTLs. Several of the genes are associated with ethylene and ABA signaling and suggest a potential linkage of o2 endosperm modification with programmed cell death. PMID- 18427772 TI - [Interaction between human chondrocytes and extracellular matrix in vitro: a contribution to autologous chondrocyte transplantation]. AB - BACKGROUND: Autologous chondrocyte transplantation (ACT) has had reasonable success for repairing small articular cartilage defects. A limiting factor for ACT is, however, the in vitro cultivation of chondrocytes because it leads to dedifferentiation. Therefore, the goal of this work was to optimize the monolayer culture of chondrocytes in vitro. MATERIAL AND METHOD: Human articular chondrocytes were plated on either collagen type II or untreated surfaces. The cells were evaluated morphologically and with immunoblotting. RESULTS: On collagen type II surfaces, a stable chondrogenic phenotype, expression of beta1 integrin, and a significant activation of phosphorylated intracellular proteins and the adaptor protein Shc could be observed up to day 20 in culture. Treatment with beta1 integrin antibody led to a loss of cell adhesion (82%). The results indicate that on collagen type II, beta1-integrin receptors are activated. Through the activation of Shc, these stimulate the Ras-MAPK pathway, which stabilizes the chondrogenic phenotype. CONCLUSION: Our results provide a practical and low-cost solution for improved long-term chondrocyte cultivation, thus providing a new perspective for using ACT on larger or arthrotic cartilage defects. PMID- 18427773 TI - [Influence of adjuvant pain medication on quality of life in the treatment of postmenopausal osteoporosis]. AB - AIM OF THE STUDY: Chronic pain is the main symptom of postmenopausal osteoporosis. This can decrease mobility and quality of life of the patients. The hypothesis of this study was that administration of an adjuvant pain medication is essential additionally to the basic therapy. The second question was if a recommendation can be formulated whether a peripheral or a central acting pain medication is more effective to prevent osteoporosis induced chronic pain. METHODS: Three pseudorandomised patient groups were prospectively compared. Group 1 was treated with alendronate, vitamin D, and calcium. Group 2 also received ibuprofen, and group 3 also received tramadol. In 117 women suffering from postmenopausal osteoporosis, quality of life was measured before and 26 weeks after therapy using the International Osteoporosis Foundation Qualeffo-41 score, and pain intensity was measured using a visual analogue scale. RESULTS: No therapy-associated complications were observed during the study. After 26 weeks, quality of life significantly increased in groups 2 and 3 compared with group 1 (p<0.001). Pain intensity decreased in group 1 by only 6 points, whereas it decreased in group 2 by 31 points and in group 3 by 24 points. Pain relief was significantly different between the treatment groups and the control group and between the treatment groups themselves (p<0.001 and p<0.01). CONCLUSION: We conclude that pain therapy with an almost peripherally acting drug such as ibuprofen can reduce osteoporosis-associated chronic pain better than a centrally acting pain medication such as tramadol. It therefore can be recommended to prescribe ibuprofen rather than tramadol for treating osteoporosis-associated chronic pain in postmenopausal women if the specific risk for gastrointestinal side effects is considered. PMID- 18427774 TI - Prone positioning in acute respiratory distress syndrome: a multicenter randomized clinical trial. AB - OBJECTIVE: We examined the effect on survival of prone positioning as an early and continuous treatment in ARDS patients already treated with protective ventilation. DESIGN AND SETTING: Open randomized controlled trial in 17 medical surgical ICUs. PATIENTS: Forty mechanically ventilated patients with early and refractory ARDS despite protective ventilation in the supine position. INTERVENTIONS: Patients were randomized to remain supine or be moved to early (within 48[Symbol: see text]h) and continuous (> or = 20 h/day) prone position until recovery or death. The trial was prematurely stopped due to a low patient recruitment rate. MEASUREMENTS AND RESULTS: Clinical characteristics, oxygenation, lung pressures, and hemodynamics were monitored. Need for sedation, complications, length of MV, ICU, and hospital stays, and outcome were recorded. PaO(2)/FIO(2) tended to be higher in prone than in supine patients after 6[Symbol: see text]h (202 +/-78 vs. 165+/-70 mmHg); this difference reached statistical significance on day 3 (234+/-85 vs. 159+/-78). Prone-related side effects were minimal and reversible. Sixty-day survival reached the targeted 15% absolute increase in prone patients (62% vs. 47%) but failed to reach significance due to the small sample. CONCLUSIONS: Our study adds data that reinforce the suggestion of a beneficial effect of early continuous prone positioning on survival in ARDS patients. PMID- 18427776 TI - Extracorporeal membrane oxygenation rescue for cardiopulmonary collapse secondary to pheochromocytoma: report of three cases. PMID- 18427775 TI - Modulation of regional nitric oxide metabolism: blood glucose control or insulin? AB - OBJECTIVE: Tight glycaemic control by intensive insulin therapy (IIT) reduces morbidity and mortality in critically ill patients. As potential mechanisms contributing to the clinical benefits we hypothesized that glycaemic control affects regional nitric oxide (NO) bioavailability by changing NO synthases (NOS) activity, NOS transcription, NOS substrate availability or the endogenous NOS inhibitor asymmetric dimethylarginine (ADMA) levels. DESIGN: Prospective, randomized experimental study. SETTING: University medical laboratory. INTERVENTIONS: In a rabbit model of prolonged critical illness we assessed the relative impact of maintaining normal insulin/normoglycaemia (n = 8), high insulin/normoglycaemia (n = 8), normal insulin/hyperglycaemia (n = 9) and high insulin/hyperglycaemia (n = 8) plasma levels over 7 days on activity and gene expression of endothelial and inducible NOS isoforms in muscle, liver and aorta biopsies, and on plasma levels of NO, arginine and ADMA. MEASUREMENTS AND RESULTS: Compared with normoglycaemic groups, both hyperglycaemic groups revealed 53% higher day-3 NO plasma levels (p < 0.05), 40% lower NOS activity in muscle (p < 0.01) and 35% lower endothelium-mediated relaxation of aortic rings (p < 0.01), 515% higher gene expression of iNOS in muscle (p < 0.01) and 99% higher eNOS gene expression in aorta (p < 0.01). Only the hyperglycaemic/hyperinsulinaemic group showed lower arginine plasma levels (53% lower, p < 0.0001). Compared with healthy controls, normoglycaemic animals revealed 33% lower ADMA levels (p < 0.05). CONCLUSIONS: In this animal model of prolonged critical illness, maintaining normoglycaemia, and not glycaemia-independent actions of insulin, prevented excessive systemic NO release on day 3 and appeared to preserve local endothelial function. Factors contributing to this finding may comprise direct endothelial cell damage, direct effects on the enzyme activity, decreased substrate availability or less NO-induced inhibition. PMID- 18427777 TI - Bi-ventricular failure following methadone overdose. PMID- 18427778 TI - End-of-life decision in a paediatric intensive care unit: decision making in light of the parents' religious beliefs. PMID- 18427779 TI - Systolic pressure variation and pulse pressure variation during modifications of arterial pressure. AB - OBJECTIVE: This study was performed to investigate the effect of vasopressor therapy on systolic pressure variation (SPV) and pulse pressure variation (PPV) compared to experimentally measured left ventricular stroke volume variation (SVV). DESIGN AND SETTING: Prospective study in a university laboratory. SUBJECTS: Twelve anesthetized and mechanically ventilated pigs. INTERVENTIONS: Increase in mean arterial pressure (by 100%) using phenylephrine and decrease (by 38%) using adenosine. MEASUREMENTS AND RESULTS: SPV and PPV were calculated and compared to SVV derived from aortic blood flow measurements. SPV was significantly affected by changes in arterial pressure [4.6% (1.5) vs. 6.3% (2.1), p < 0.05, increased vs. decreased arterial pressure], whereas PPV did not change during modifications of arterial pressure. During baseline conditions and decreased afterload, correlation with SVV was good both for SPV (r =0.892 and r = 0.859, respectively) and for PPV (r = 0.870 and r = 0.871, respectively) (all p < 0.001). Correlation with SVV was only moderate during increased arterial pressure (r = 0.683 for SPV and r = 0.732 for PPV, p < 0.05). CONCLUSION: For guiding fluid therapy in patients under vasopressor support, PPV seems superior to SPV. PMID- 18427780 TI - Variability in the concentrations of intravenous drug infusions prepared in a critical care unit. AB - OBJECTIVE: To quantify the variability in the concentration of drug infusions prepared on an intensive care unit and establish whether there was a relationship between the quality of syringe labeling and drug preparation. DESIGN: Audit carried out over 3 weeks in May 2006 and completed in May 2007. SETTING: The adult neurosciences critical care unit of a UK university teaching hospital. INTERVENTIONS: Daily collections of discarded syringes containing midazolam, insulin, norepinephrine, dopamine, potassium or magnesium. MEASUREMENTS AND RESULTS: Residual solutions in the syringes were sampled and the concentrations measured. Syringe labels were inspected and awarded a score for labeling quality based on an 11-point scale. A total of 149 syringes were analyzed. Six of the magnesium syringes contained 4-5 times too much Mg(2+), presumably because of confusion about converting millimoles to grams. The majority of the other infusions differed from the expected concentration by more than 10%. Magnesium infusions were least likely to be properly labeled (p= 0.012), and there was a positive correlation between quality of syringe labeling and drug preparation (p=0.002). After the introduction of a new electrolyte prescription chart, magnesium and potassium preparation significantly improved but there was still substantial variability. CONCLUSIONS: These findings present a strong argument for the use of pre-prepared syringes or standardized drug preparation and labeling systems. They also highlight once again the difficulties healthcare professionals encounter when dealing with different ways of expressing drug concentrations. PMID- 18427782 TI - The application of risk-adjusted control charts using the Paediatric Index of Mortality 2 for monitoring paediatric intensive care performance in Australia and New Zealand. AB - OBJECTIVE: To assess the applicability of risk-adjusted sequential control charts using the Paediatric Index of Mortality version 2 for monitoring of the quality of paediatric intensive care. DESIGN: Observational study. SETTING: A registry of patient admissions to paediatric intensive care units (PICUs) in Australia and New Zealand. PATIENTS: A total of 10,710 patients admitted to eight PICUs during a 24-month period. MEASUREMENTS AND RESULTS: A series of risk-adjusted control charts was created for each PICU. Modified sequential probability ratio tests were used to test the hypothesis that the PICUs being monitored were 'out of control', where loss of control was arbitrarily defined as the odds of death exceeding twice the odds of dying as estimated by PIM2. In 24 months of monitoring, there was one alarm signal, suggesting the odds of deaths had doubled, and there was one signal, in another PICU, suggesting the odds of death had halved. CONCLUSIONS: The major advantage of risk-adjusted sequential control charts is that the technique allows unit performance to be monitored continuously over time, rather than intermittently, with the aim of rapidly detecting a change in performance as soon as possible after it occurs. This technique is suitable for continuously screening for a change in outcome within a PICU over time and complements other methods of monitoring the quality of paediatric intensive care. PMID- 18427781 TI - Understanding organ dysfunction in hemophagocytic lymphohistiocytosis. AB - OBJECTIVE: This review aims to help critical care clinicians maintain a high level of suspicion regarding the diagnosis of Hemophagocytic Histiolymphocytosis (HLH). It describes the clinical and laboratory features of HLH, outlines its pathophysiology and reviews the most frequent etiologies related to HLH. Prognostic factors and therapeutic options are also reported. DATA SOURCES: Review of the literature. RESULTS: The diagnosis of HLH relies on the association of clinical abnormalities and hemophagocytosis in bone marrow, spleen, or lymph node specimens. Liver, pulmonary, renal, cardiac and skin involvement may occur at various degrees possibly leading to multiple organ failure. Three main etiologies can be found, namely infections, lymphoproliferative diseases, or connective tissue diseases. Immune deficiency is often retrieved. Mortality can be as high as 50%. Although clinically mimicking severe sepsis, HLH has a distinct pathophysiology on which specific therapy is based. Early diagnosis and treatment is mandatory to increase the chances of survival. CONCLUSION: The comprehensive management of severe HLH requires the involvement of a multidisciplinary team in order to determine the best therapeutic strategy and to identify the underlying cause. PMID- 18427783 TI - Expression of nuclease gene nucA, a member of an operon putatively involved in uracil removal from DNA and its subsequent reuse in Prevotella bryantii. AB - The genomic region of Prevotella bryantii TC1-1 that conferred an increased nucleolytic activity on Escherichia coli was characterized. It contains two divergent transcriptional units separated by an AT-rich promoter region. One unit is comprised of three genes involved in nucleotide metabolism. nucA, the first gene of this unit, whose product belongs to exonuclease/endonuclease/phosphatase Pfam family, was thought to be required for the increased nucleolytic activity and various expression strategies were employed to confirm its role. The nucA expression was only successful in cell free system where DNase and RNase activity was observed. Two genes downstream of nucA code for a putative uracil DNA glycosylase and uridine kinase which could be involved in the removal of misincorporated uracil from DNA and its reuse. Given that apurinic/apyrimidinic nuclease activity is required after uracil removal from DNA, it was somewhat surprising to find out that nucA, whose product belongs to protein family consisting mostly of apurinic/apyrimidinic nucleases, has no apurinic/apyrimidinic activity. PMID- 18427784 TI - Subchronic and chronic PCP treatment produces temporally distinct deficits in attentional set shifting and prepulse inhibition in rats. AB - RATIONALE: We have previously demonstrated that subchronic (five daily administrations of 2.6 mg/kg PCP) and chronic intermittent administration of 2.6 mg/kg PCP to rats produces hypofrontality and other neurochemical changes akin to schizophrenia pathology (Cochran et al., Neuropsychopharmacology, 28:265-275, 2003). OBJECTIVES: We sought to determine whether behavioral alterations related to discrete aspects of schizophrenia are also induced by these PCP treatment regimes. MATERIALS AND METHODS: Following administration of vehicle or PCP according to the protocols described above, rats were assessed for attentional set shifting ability, prepulse inhibition (PPI), or social interaction and the locomotor response to a challenge dose of amphetamine. RESULTS: Ability to shift attentional set was impaired 72 h after the last PCP administration following the subchronic and chronic intermittent treatment regimes. PPI was disrupted after each acute administration of PCP in animals under the subchronic treatment regime. However, PPI deficits were not sustained 72 h after the last of five daily administrations. In subchronic and chronic PCP treated animals, no change was found in social interaction behavior, and there was little change in baseline or amphetamine-stimulated locomotor activity, employed as an indicator of dopaminergic hyperfunction. CONCLUSIONS: The temporally distinct behavioral effects of these PCP treatment regimes suggest that PPI deficits relate directly to acute NMDA receptor antagonism, whereas the more enduring set shifting deficits relate to the longer term consequences of NMDA receptor blockade. Therefore, these subchronic and chronic PCP treatment regimes produce hypofrontality (Cochran et al., Neuropsychopharmacology, 28:265-275, 2003) and associated prefrontal cortex-dependent deficits in behavioral flexibility which mirror core deficits in schizophrenia. PMID- 18427785 TI - Towards chemical analysis of nanostructures in biofilms II: tip-enhanced Raman spectroscopy of alginates. AB - This study examines the feasibility of using tip-enhanced Raman spectroscopy (TERS) for label-free chemical characterization of nanostructures in biological systems. For this purpose, a well-defined model system consisting of calcium alginate fibers is studied. In a companion paper, calcium alginate fibers and their network structures were shown to be a good model for the extracellular polysaccharides of biofilms at the nanoscale. TERS analysis of biological macromolecules, such as alginates, is complicated by heterogeneity in their sequence, molecular weight, and conformations, their small Raman cross-section, and the large number of functional groups, which can chemically interact with the silver surface of the tip and cause significant band shifts. Due to these effects, Raman frequencies in TERS spectra of biopolymers do not necessarily resemble band positions in the normal Raman spectrum of the bulk material, as is the case for less complex samples (e.g., dye molecules) studied so far. Additionally, analyte decomposition due to laser heating can have a significant influence, and carbon contamination signals can sometimes even overwhelm the weak analyte signals. Based on the investigation of alginates, strategies for spectra correction, choice of appropriate reference samples, and data interpretation are presented. With this approach, characteristic frequency ranges and specific marker bands can be found for biological macromolecules that can be employed for their identification in complex environments. PMID- 18427786 TI - Towards chemical analysis of nanostructures in biofilms I: imaging of biological nanostructures. AB - Due to their direct influence on the stability of bacterial biofilms, a better insight into the nanoscopic spatial arrangement of the different extracellular polymeric substances (EPS), e.g., polysaccharides and proteins, is important for the improvement of biocides and for process optimization in wastewater treatment and biofiltration. Here, the first application of a combination of confocal laser scanning microscopy (CLSM) and atomic force microscopy (AFM) to the investigation of river-water biofilms and related biopolymers is presented. AFM images collected at selected areas of CLS micrographs dramatically demonstrate the heterogeneity of biofilms at the nanometer scale and the need for a chemical imaging method with nanoscale resolution. The nanostructures (e.g., pili, flagella, hydrocolloids, and EPS) found in the extracellular matrix are classified according to shape and size, which is typically 50-150 nm in width and 1-10 nm in thickness, and sets the demands regarding spatial resolution of a potential chemical imaging method. Additionally, thin layers of the polysaccharide alginate were investigated. We demonstrate that calcium alginate is a good model for the EPS architecture at the nanometer scale, because of its similar network-like structure. PMID- 18427787 TI - Highly sensitive restriction enzyme assay and analysis: a review. AB - Biological assays at the single molecule level are crucial to fundamental studies of DNA-protein mechanisms. In order to cater for high throughput applications, one area of immense research potential is single-molecule bioassays where miniaturized devices are developed to perform rapid and effective biological reactions and analyses. With the success of various emerging technologies for engineering miniaturized structures down to the nanoscale level, supported by specialized equipment for detection, many investigations in the field of life science that were once thought impossible can now be actively explored. In this review, the significance of downscaling to the single-molecule level is firstly presented in selected examples, with the focus placed on restriction enzyme assays. To determine the effectiveness of single-molecule restriction enzyme reactions, simple and direct analytical methods based on DNA stretching have often been reliably employed. DNA stretching can be realized based on a number of working principles related to the physical forces exerted on the DNA samples. We then discuss two examples of a nanochannel system and a microchamber system where single-molecule restriction enzyme digestion and DNA stretching have been integrated, which possess prospective capabilities of developing into highly sensitive and high-throughput restriction enzyme assays. Finally, we take a brief look at the general trends in technological development in this field by comparing the advantages and disadvantages of performing assays at bulk, microscale and single-molecule levels. PMID- 18427788 TI - Determination of immunomodulatory effects: focus on functional analysis of phagocytes as representatives of the innate immune system. AB - The evaluation of the effects of drugs or chemicals on the functions of the immune system is an increasingly important task. Due to the accessibility of primary cells and cell lines, in vitro cellular functional tests are frequently being performed with cells representing the innate immune system, in particular those with phagocytotic activities, such as neutrophils and macrophages. Suitable functional parameters are the efficiency of phagocytosis, the efficiency with which viable pathogens are killed, the production of reactive oxygen and nitrogen species (ROS and RNS) and that of cytokines. Corresponding analytical procedures are available, but standardization is required, as varying the procedure may influence the outcomes of the assays. PMID- 18427789 TI - Heteroatom(isotope)-tagged proteomics via ICP-MS: screening and quantification of proteins and their post-translational modifications. PMID- 18427790 TI - Whither "quant"? An examination of the curriculum and testing methods for quantitative analysis courses taught in universities and colleges in the Western USA. PMID- 18427791 TI - Grasping numbers. AB - Both theoretical and empirical studies suggest that numerical processing is intimately linked to representations of goal-directed hand actions. Further evidence for this possibility is provided here by the results of two experiments, both of which revealed a powerful influence of numerical magnitude on the selection of hand grasping movements. Human participants performed either power or precision grip responses based on the semantic properties (e.g., parity) of visual Arabic numerals, in Experiment 1, or depending on their surface characteristics (e.g., colour), in Experiment 2. In both the experiments, it was found that small numerical values facilitated precision grip (commonly used to grasp small objects), while large numerical value potentiated power grip (commonly used to grasp large objects). These findings reveal that perceiving numbers can automatically prime grasping gestures, in a similar manner to viewing physical objects. This result is coherent with the view that processing of symbolic (numerical) and physical quantitative information converges onto a shared magnitude system representing the coordinates of action. PMID- 18427792 TI - Paired-pulse behavior of visually evoked potentials recorded in human visual cortex using patterned paired-pulse stimulation. AB - Paired-pulse stimulation techniques are used as common tools to investigate cortical excitability and cortical plastic changes. Similar to investigations in the somatosensory and motor system here we applied a new paired-pulse paradigm to study the paired-pulse behavior of visually evoked potentials (VEPs) in 25 healthy subjects. VEPs were recorded and the responses to the first and the second P100 peak were analyzed at different SOAs [stimulus onset asynchrony (SOA) = interstimulus interval (ISI) + pulse duration (13 ms)]. Two measures describe the paired pulse interaction: the "amplitude ratio", the ratio of the second to the first amplitude, and the "latency shift", the difference of the inter-peak interval between the P100 peaks and the respective SOA. To separate alterations in the amplitude of the second VEP response due to changes in paired-pulse inhibition from those originating from superposition of the two waveforms, particularly at short SOAs, we created a waveform template from recordings made at SOAs of 1 s, where interaction can be assumed to be negligible. Superposed traces of VEP recordings were then created by adding two templates at delays corresponding to the SOAs used. The original recordings were then digitally subtracted from the traces obtained by superposition. Analysis of the subtracted traces revealed evidence that at short SOAs the second VEP response is substantially suppressed, a finding comparable to the paired-pulse inhibition described for motor and somatosensory cortex following paired-pulse stimulation. However, paired-pulse inhibition seen in V1 varied considerably from subject to subject, both in respect to amplitude, and to time of maximal inhibition. We found paired-pulse inhibition ranging from 12 to 76% (mean 34%) at SOAs between 80 (shortest discriminable SOA) and 320 ms (mean 128 ms). At intermediate SOAs between 80 and 720 ms (mean 215 ms) the amplitude ratios were between 94 and 145% (mean 116%) indicative of slight paired-pulse facilitation. Comparable to recovery studies by means of paired-pulse median nerve stimulation in somatosensory cortex, at shorter SOAs we found a delayed second VEP response. Combined together, our findings suggest that VEPs are characterized by significant paired-pulse inhibition at short SOAs, a phenomenon reminiscent of findings reported in other modalities. Possible mechanisms and pharmacological properties of the described paired-pulse behavior in visual cortex remain to be explored. PMID- 18427793 TI - Soleus H-reflex excitability during pedaling post-stroke. AB - A major contributor to impaired locomotion post-stroke is abnormal phasing of paretic muscle activity, but the mechanisms remain unclear. Previous studies have shown that, in the paretic limb of people post-stroke, Group Ia reflexes are abnormally elevated and fail to decrease in amplitude during locomotion. Hence, we hypothesized that inappropriate muscle phasing may be associated with enhanced transmission in the monosynaptic Group Ia afferent pathway. Soleus (SO) H reflexes were used to examine transmission in the Group Ia afferent pathway to SO motor neurons during pedaling, a locomotor task in which abnormal muscle phasing is evident. Our hypothesis predicted that H-reflexes would be elevated during the flexion phase of pedaling where inappropriate SO activity occurs. H-reflexes were elicited in paretic (n = 13) and neurologically intact (NI, n = 26) individuals at 11 different positions in the pedaling cycle and during tonic plantar flexion at comparable limb positions and levels of SO EMG. In both groups, SO H-reflexes were smaller during pedaling as compared to matched tonic plantar flexion. In the NI group, but not the paretic group, SO H-reflex amplitude was significantly modulated across the pedaling cycle. H-reflexes were large during extension and small during flexion. Reduced H-reflex modulation post-stroke was associated with the level of neuromuscular impairment as indicated by Fugl-Meyer score. However, regardless of impairment level, stroke subjects displayed H-reflex suppression during the flexion phase of pedaling. After correcting for the level of background muscle activity, H-reflexes were found to be larger in paretic as compared to NI individuals, regardless of the phase of the pedaling cycle. We conclude that Group Ia afferent transmission is enhanced in the paretic SO of people post-stroke as compared to NI individuals. However, contrary to our hypothesis, enhanced transmission in the Group Ia monosynaptic spinal pathway is not specifically associated with extraneous extensor muscle activity during the flexion phase of pedaling and is unlikely to account for abnormal locomotor muscle phasing post-stroke. This result is important because it suggests that, despite the presence of hyperactive monosynaptic reflexes post-stroke, this impairment may not make an important contribution to abnormal locomotor muscle activity. PMID- 18427794 TI - Online corrections can produce illusory bias during closed-loop pointing. AB - This experiment examined whether the impact of pictorial illusions during the execution of goal-directed reaching movements is attributable to ocular motor signaling. We analyzed eye and hand movements directed toward both the vertex of the Muller-Lyer (ML) figure in a closed-loop procedure. Participants pointed to the right vertex of a visual stimulus in two conditions: a control condition wherein the figure (in-ML, neutral, out-ML) presented at response planning remained unchanged throughout the movement, and an experimental condition wherein a neutral figure presented at response planning was perturbed to an illusory figure (in-ML, out-ML) at movement onset. Consistent with previous work from our group (Heath et al. in Exp Brain Res 158:378-384, 2004; Heath et al. in J Mot Behav 37:179-185, 2005b), action-bias present in both conditions; thus illusory bias was introduced into during online control. Although primary saccades were influenced by illusory configurations (control conditions; see Binsted and Elliott in Hum Mov Sci 18:103-117, 1999a), illusory bias developed within the secondary "corrective" saccades during experimental trials (i.e., following a veridical primary saccade). These results support the position that a unitary spatial representation underlies both action and perception and this representation is common to both the manual and oculomotor systems. PMID- 18427795 TI - Arguments against the proposed randomised trial (ARUBA). PMID- 18427796 TI - ARUBA--beating natural history in unruptured brain AVMs by intervention. PMID- 18427797 TI - Endovascular snare kit in the combined antegrade and retrograde management of ureteral avulsion: report of two cases. AB - We report the feasible and safe use of the Amplatz Goose Neck Snare kit for avulsed ureter retrieval during ureteroscopy. A 49-year-old lady and a 61-year old man complaining of urolithiasis underwent ureteroscopy; following stone fragmentation, and basketing avulsion of the ureter occurred. Using the Amplatz Goose Neck Snare kit it was possible to place an indwelling ureteral catheter in both cases aiming at restoring the urinary upper tract continuity. The snare assisted endovascular technique may be an interesting tool even in endourology for the management of ureteral avulsion. This endoscopic mini-invasive procedure makes it possible to avoid an immediate invasive surgical approach often resulting in nephrectomy, having time for planning a possible durable conservative treatment. PMID- 18427798 TI - Relaxivities of paramagnetic liposomes: on the importance of the chain type and the length of the amphiphilic complex. AB - Nuclear magnetic relaxation dispersion (NMRD) profiles of unilamellar DPPC liposomes incorporating Gd-DTPA-bisamides with alkyl chains of 12 to 18 C atoms in their external and internal layers were recorded in order to study the influence that the chain length and structure of Gd-bisamides incorporated in the liposomal membrane have on their proton relaxivity. The NMRD profiles recorded at 310 K show that the relaxivity reaches a minimum value when the carbon chain lengths of the phospholipid and of the Gd complex match and is at a maximum in the presence of a carbon-carbon double bond. For these DPPC paramagnetic liposomes, the longer the aliphatic chains of the complex, the larger will be its immobilization in the membrane. In addition, the presence of an unsaturated carbon-carbon bond in the alkyl chain of the Gd complex induces an increase of its mobility and of its water exchange rate with, as a result, a much greater efficiency as an MRI contrast agent. PMID- 18427799 TI - A model for thermal exchange in axons during action potential propagation. AB - Several experiments have shown that during propagation of the action potential in axons, thermal energy is locally exchanged. In this paper, we use a simple model based on statistical physics to show that an important part of this exchange comes from the physics of the effusion. We evaluate, during the action potential propagation, the variation of internal energy and of the energy associated with the chemical potential of the effusion of water and ions to extract the thermal energy exchanged. The temperature exchanged is then evaluated on the area where the action potential is active. Results give a good correspondence between experimental work and this model, showing that an important part of the thermal energy exchange comes from the statistical cooling power of the effusion. PMID- 18427800 TI - Arrangement of ceramide [EOS] in a stratum corneum lipid model matrix: new aspects revealed by neutron diffraction studies. AB - The lipid matrix in stratum corneum (SC) plays a key role in the barrier function of the mammalian skin. The major lipids are ceramides (CER), cholesterol (CHOL) and free fatty acids (FFA). Especially the unique-structured omega-acylceramide CER[EOS] is regarded to be essential for skin barrier properties by inducing the formation of a long-periodicity phase of 130 angstroms (LPP). In the present study, the arrangement of CER[EOS], either mixed with CER[AP] and CHOL or with CER[AP], CHOL and palmitic acid (PA), inside a SC lipid model membrane has been studied for the first time by neutron diffraction. For a mixed CER[EOS]/CER[AP]/CHOL membrane in a partly dehydrated state, the internal membrane nanostructure, i.e. the neutron scattering length density profile in the direction normal to the surface, was obtained by Fourier synthesis from the experimental diffraction patterns. The membrane repeat distance is equal to that of the formerly used SC lipid model system composed of CER[AP]/CHOL/PA/ChS. By comparing both the neutron scattering length density profiles, a possible arrangement of synthetic long-chain CER[EOS] molecules inside a SC lipid model matrix is suggested. The analysis of the internal membrane nanostructure implies that one CER[EOS] molecule penetrates from one membrane layer into an adjacent layer. A 130 angstroms periodicity phase could not be observed under experimental conditions, either in CER/CHOL mixtures or in CER/CHOL/FFA mixture. CER[EOS] can be arranged inside a phase with a repeat unit of 45.2 angstroms which is predominately formed by short-chain CER[AP] with distinct polarity. PMID- 18427801 TI - Inter-subunit disulfide cross-linking in homomeric and heteromeric P2X receptors. AB - P2X receptors are ATP-gated cation channels and assembled as homotrimers or heterotrimers from seven cloned subunits. Each subunit contains two transmembrane domains connected by a large extracellular loop. We have previously shown that replacement of two conserved residues, K68 and F291, by cysteine residues leads to disulfide cross-linking between neighbouring P2X(1) subunits. Since mutation of these residues results in a reduced ATP potency and cysteine cross-linking is prevented in the presence of ATP, we suggested an inter-subunit ATP binding site. To investigate whether the proximity of these residues is preserved in other P2X subtypes, we tested for spontaneous cystine formation between the corresponding P2X(2 )(K69C, F289C), P2X(3 )(K63C, F280C), and P2X(4 )(K67C, F294C) mutants upon pairwise expression in Xenopus laevis oocytes. Non-reducing SDS-PAGE analysis of the purified receptors revealed a specific dimer formation between P2X(2)K69C and P2X(2)F289C mutants. Likewise, co-expression of P2X(1)K68C and P2X(2)F289C, but not P2X(1)F291C and P2X(2)K69C, mutants resulted in dimer formation between the respective subunits. Cross-linked P2X(1/2) heteromers showed strongly reduced or absent function that was selectively recovered upon treatment with DTT. Cross linking was less efficient between P2X(3) or P2X(4) mutants but could be enhanced by the short cysteine-reactive cross-linker MTS-2-MTS. These results show that the spatial proximity and/or orientation of residues analogous to positions K68 and F291 in P2X(1) are preserved in P2X(2) receptors and at one of two possible interfaces in heteromeric P2X(1/2) receptors but appears to be redundant for P2X(3) and P2X(4) receptor function. PMID- 18427802 TI - The role of protein-solvent hydrogen bond dynamics in the structural relaxation of a protein in glycerol versus water. AB - We used MD simulations to investigate the dependence of the dynamics of a soluble protein, RNase A, on temperature and solvent environment. Consistent with neutron scattering data, the simulations predict that the protein undergoes a dynamical transition in both glycerol and aqueous solutions that is absent in the dry protein. The temperature of the transition is higher, while the rate of increase with temperature of the amplitudes of motion on the 100 ps timescale is lower, in glycerol versus water. Analysis of the dynamics of hydrogen bonds revealed that the protein dynamical transition is connected to the relaxation of the protein solvent hydrogen bond network, which, in turn, is associated with solvent translational diffusion. Thus, it appears that the role of solvent dynamics in affecting the protein dynamical transition is qualitatively similar in water and glycerol. PMID- 18427803 TI - Efficient production of active form of recombinant cassava hydroxynitrile lyase using Escherichia coli in low-temperature culture. AB - Overexpression and production of the high concentration of hydroxynitrile lyase from cassava (Manihot esculenta (MeHNL, EC 4.1.2.39)) were investigated. Hydroxynitrile lyase is a useful enzyme for the production of optically active cyanohydrin compounds. The production of MeHNL was increased by changing the rare codons of the original sequence of cassava MeHNL. However, most of the produced MeHNL was in the insoluble form. In order to increase the solubility of MeHNL, the effects of the cultivation temperature were investigated. When the cultivation temperature was reduced, the cell yield and the ratio of soluble MeHNL increased significantly. The enzyme activity and yield at low-temperature cultures (17 degrees C) were 850 times higher than those obtained at the optimum growth temperature of 37 degrees C. The rate of MeHNL production in the present study was calculated as 3,000 unit/h. Low-temperature cultivation was very effective in improving the productivity of the active form of MeHNL. Unlike the temperature-shift method, low-temperature cultivation has more potential for the large-scale production of MeHNL for the optically active cyanohydrin production. PMID- 18427804 TI - The yeast Kluyveromyces marxianus and its biotechnological potential. AB - Strains belonging to the yeast species Kluyveromyces marxianus have been isolated from a great variety of habitats, which results in a high metabolic diversity and a substantial degree of intraspecific polymorphism. As a consequence, several different biotechnological applications have been investigated with this yeast: production of enzymes (beta-galactosidase, beta-glucosidase, inulinase, and polygalacturonases, among others), of single-cell protein, of aroma compounds, and of ethanol (including high-temperature and simultaneous saccharification fermentation processes); reduction of lactose content in food products; production of bioingredients from cheese-whey; bioremediation; as an anticholesterolemic agent; and as a host for heterologous protein production. Compared to its congener and model organism, Kluyveromyces lactis, the accumulated knowledge on K. marxianus is much smaller and spread over a number of different strains. Although there is no publicly available genome sequence for this species, 20% of the CBS 712 strain genome was randomly sequenced (Llorente et al. in FEBS Lett 487:71-75, 2000). In spite of these facts, K. marxianus can envisage a great biotechnological future because of some of its qualities, such as a broad substrate spectrum, thermotolerance, high growth rates, and less tendency to ferment when exposed to sugar excess, when compared to K. lactis. To increase our knowledge on the biology of this species and to enable the potential applications to be converted into industrial practice, a more systematic approach, including the careful choice of (a) reference strain(s) by the scientific community, would certainly be of great value. PMID- 18427805 TI - Construction and characterization of an enhanced GFP-tagged anti-BAFF scFv antibody. AB - Elevated levels of B-cell-activating factor of the TNF family (BAFF) have been implicated in the pathogenesis of autoimmune diseases in human. In this study, an anti-BAFF single-chain antibody fragment (scFv) was genetically linked to the C terminus of the enhanced green fluorescent protein (EGFP) to generate an EGFP/scFv fusion protein. The EGFP/scFv fusion protein had an apparent molecular weight of 52 kDa and was identified by sodium dodecyl sulfate polyacrylamide gel electrophoresis and Western blot analysis. After being purified by immobilized metal affinity chromatography, the fusion protein exhibited similar fluorescence spectra with native EGFP. Enzyme-linked immunosorbent assay and fluorescence activated cell sorting showed the EGFP/scFv could bind to human soluble BAFF and BAFF positive cell lines in vitro. The binding of EGFP/scFv can also be visualized under laser scanning confocal microscopy. Furthermore, the results of the competition assay indicated its antigen binding specificity. Therefore, the fusion protein EGFP/scFv has several characteristics including high sensitivity, stability and convenience for manipulation, and can be a powerful tool for the study of the underlying pathology of BAFF relevant to autoimmune diseases. PMID- 18427806 TI - Purification and characterization of organic solvent stable protease from Bacillus licheniformis RSP-09-37. AB - A protease was purified from the cell-free supernatant of Bacillus licheniformis RSP-09-37, a mutant from a thermophilic bacterial strain, B. licheniformis RSP 09, using affinity chromatography with alpha-casein agarose resin. The protease was purified 85-fold to electrophoretic homogeneity. The apparent molecular mass of purified protease was 55 kDa using gel filtration in high-performance liquid chromatography, which is in agreement with the results obtained from sodium dodecyl sulfate-polyacrylamide gel electrophoresis, suggesting a monomeric nature of the protein. The purified protease revealed temperature optima of 50 degrees C and pH optima of 10.0 and was classified as serine protease based on its complete inhibition with phenyl methyl sulfonyl fluoride. The purified protease exhibited tolerance to both detergents and organic solvent. The synthetic activity of the protease was tested using the transesterification reaction between N-acetyl-L phenylalanine-ethyl ester and n-propanol in organic solvents varying in their log P values and the kinetic parameters of the enzyme in these organic solvents were studied. The enzyme has potential to be employed for synthetic reactions and in detergent formulations. PMID- 18427807 TI - Long-term evaluation of renal toxicity after peptide receptor radionuclide therapy with 90Y-DOTATOC and 177Lu-DOTATATE: the role of associated risk factors. AB - PURPOSE: Peptide receptor radionuclide therapy (PRRT) of neuroendocrine tumours with (90)Y-DOTATOC and (177)Lu-DOTATATE is promising. The kidney is the critical organ and despite renal protection, function loss may become evident years later. The aim of this study was to analyse renal parameters in patients who had undergone dosimetry before PRRT. METHODS: Among those in protocols at our institution, 28 patients were considered: 23 received (90)Y-DOTATOC (3.8-29.2 GBq, median 12.2) and five received (177)Lu-DOTATATE (20.7-29.2 GBq, median 23.2). Patients were followed up after therapy for creatinine and creatinine clearance loss (CCL) for 3-97 months (median 30). Renal doses and bio-effective doses (BED) were calculated (MIRD, LQ model). RESULTS: After (90)Y-DOTATOC toxicity on creatinine according to NCI criteria occurred in nine cases (seven grade 1, one grade 2, one grade 3), CCL at 1 year was >5% in 12 cases and >10% in eight. A 28-Gy BED threshold was observed in patients with risk factors (mainly hypertension and diabetes), while it was 40 Gy in patients without risk factors. Probably due to the low number of patients, despite the absence of severe toxicity after hyper-fractionated PRRT, clear correlations between fractionation and toxicity could not be found. After (177)Lu-DOTATATE, no toxicity occurred in 1-2 year follow-up; CCL at 1 year >5% occurred in three patients and >10% in two. CONCLUSIONS: Our results indicate the importance of clinical screening for risk factors: In this case, a BED <28 Gy is recommended. Fractionation of therapy is important in order to decrease toxicity, and further studies are needed to evaluate its clinical impact. PMID- 18427808 TI - Malaria detection with the Sysmex XE-2100 hematology analyzer using pseudoeosinophilia and abnormal WBC scattergram. AB - Recent investigation using the Sysmex XE-2100 hematology analyzer (Sysmex Corporation, Japan) has demonstrated erroneously high eosinophil counts and abnormal white blood cell (WBC) scattergrams in malaria cases. This study was conducted to assess the diagnostic efficiency of the Sysmex XE-2100 analyzer for malaria. One hundred forty-four patients initially diagnosed with Plasmodium vivax infection, 319 patients with febrile illness, and 24 patients who underwent malaria treatment were analyzed. Atypical features on Sysmex XE-2100 analyzer were categorized as pseudoeosinophilia (a gap of more than 5% in eosinophil counts between the Sysmex XE-2100 analyzer and microscopic examination) and abnormal WBC scattergram. Pseudoeosinophilia or abnormal WBC scattergram were detected in 100 of 144 malaria-positive samples (sensitivity 69.4%, specificity 100%). The samples with pseudoeosinophilia or abnormal WBC scattergrams showed significantly higher parasite counts than the samples without pseudoeosinophilia or an abnormal WBC scattergram (P<0.05). All 24 samples from patients for whom the malaria smear was repeated after malaria treatment was initiated showed a normalized eosinophil count and a normal WBC histogram. In conclusion, attention to differential count and WBC scattergrams provided by the Sysmex XE-2100 would be a valuable tool in malaria detection. PMID- 18427809 TI - Carnitine and carnitine acetyltransferases in the yeast Saccharomyces cerevisiae: a role for carnitine in stress protection. AB - To date, the only reported metabolic and physiological roles for carnitine in Saccharomyces cerevisiae are related to the activity of the carnitine shuttle. In yeast, the shuttle transfers peroxisomal activated acetyl-residues to the mitochondria. However, acetyl-CoA can also be metabolised by the glyoxylate cycle to form succinate. The two pathways, therefore, provide a metabolic bypass for each other, and carnitine-dependent phenotypes have only been described in strains with non-functional peroxisomal citrate synthase, Cit2p. Here, we present evidence for a role of carnitine in stress protection that is independent of CIT2 and of the carnitine shuttle. Data show that carnitine improves growth during oxidative stress and in the presence of weak organic acids in wt and in CAT deletion strains. Our data also show that strains with single, double and triple deletions of the three CAT genes generally present identical phenotypes, but that the deletion of CAT2 decreases survival during oxidative stress in a carnitine independent manner. Overexpression of single CAT genes does not lead to cross complementation, suggesting a highly specific metabolic role for each enzyme. The data suggest that carnitine protects cells from oxidative and organic acid stress, while CAT2 contributes to the response to oxidative stress. PMID- 18427810 TI - Extrinsic light:dark cycles, rather than endogenous circadian cycles, affect the photoperiodic counter in the pitcher-plant mosquito, Wyeomyia smithii. AB - A wide diversity of organisms use photoperiod (daylength) as an environmental cue to anticipate the changing seasons and to time various life-history events such as dormancy and migration. Photoperiodic time measurement consists of two main components, (1) the photoperiodic timer that discriminates between long and short days, and (2) the photoperiodic counter that accumulates and stores information from the timer and then induces the phenotypic output. Herein, we use extended night treatments to show that light is necessary to accumulate photoperiodic information across the geographic range of the mosquito, Wyeomyia smithii and that the photoperiodic counter counts extrinsic (external) light:dark cycles and not endogenous (internal) circadian cycles. PMID- 18427811 TI - Surgical strategies in the management of ecthyma gangrenosum in paediatric oncology patients. AB - Clinical presentation and microbiology profiles of neutropenic paediatric oncology patients presenting with ecthyma gangrenosum (EG) were studied. Surgical strategies deployed for these critically ill children are reported. Between 1994 and 2005, all children with EG were identified. Case notes were reviewed. Hospital course and long-term outcome were documented. Ten patients were identified. Eight had acute lymphoblastic leukaemia, one child had acute myeloid leukaemia and another had rhabdomyosarcoma. Lesions occurred in the perineal region (n = 5), buttocks (n = 2), thigh (n = 2) and the face (n = 1). Seven children had positive blood cultures for Pseudomonas aeruginosa. Surgery included (1) radical debridement, and (2) debridement with covering colostomy for four of those with perianal lesions. Ecthyma gangrenosum is a rapidly spreading and potentially lethal condition. Paediatric oncology patients with neutropenia are at a high risk. Surgical excision is crucial for progressive lesions to prevent mortality. PMID- 18427812 TI - Oesophageal atresia with cleft lip and palate: a marker for associated lethal anomalies? AB - An adverse association between oesophageal atresia (OA) and cleft lip-palate (3% incidence) has been reported. The present study analyses outcomes of this rare association at a UK paediatric surgical centre. Hospital charts of newborns diagnosed with OA were reviewed. Demographics, associated anomalies and prognostic classification (after Spitz 1994) were recorded. Mortality rates and causes of death were examined in OA babies with cleft lip-palate. Of 152 patients treated for OA, five babies (3%) had cleft lip-palate. All of these newborns had common variant OA-TEF and were Spitz group II category. Deaths occurred in 3 of 5 patients (60%) in the OA-cleft group compared to only 8 of 147 patients (5%) without clefts (p < 0.005; Fisher's exact test). OA-cleft non-survivors succumbed to tetralogy of Fallot (n = 2) and trisomy 18 (n = 1; treatment withdrawn). Both survivors with cleft lip-palate had features of the VACTERL sequence: one baby also had Goldenhaar syndrome, the other aortic coarctation. These children now attend mainstream school. Although high-quality survival is possible in OA with cleft lip-palate, this rare phenotype is associated with a substantially decreased survival. Rather than causing death directly, the combination of OA and cleft lip-palate appears to be a marker for further lethal anomalies. PMID- 18427813 TI - Protective effects of vitamin E and omeprazole on the hypoxia/reoxygenation induced intestinal injury in newborn rats. AB - Evaluation of prophylactic effects of omeprazole and/or vitamin E on the formation of free oxygen radicals (FOR) and bowel histopathology in the newborn rat model of hypoxia/reoxygenation (H/R) that resembles human necrotizing enterocolitis (NEC). Eighty newborn rats were randomly divided into eight groups. H/R was done using airtight chamber. Rats were exposed to 100% CO2 for 15 min followed by a reoxygenation for the next 15 min with 100% O2. Group 1 (n = 10) was the control group. Group 2 (n = 10) rats received vitamin E. In Group 3 (n = 10) omeprazole was administrated. Group 4 (n = 10) rats received omeprazole and vitamin E. Group 5 (n = 10) rats were subjected to H/R two times for 2 days and one time for 3 days. Group 6 (n = 10) received vitamin E in addition to H/R for 5 days and in Group 7 (n = 10) omeprazole in addition to H/R for 5 days. In Group 8 (n = 10), vitamin E and omeprazole and H/R were applied for 5 days. Rats were killed at the end of the each process and bowel specimens were harvested for histopathological and biochemical investigations. We administrated vitamin E intramuscularly 300 unit/kg per day and omeprazole orally 20 mg/kg per day. Malondialdehyde (MDA), xanthine oxidase (XO), xanthine dehydogenase (XDH) and XO/(XO + XDH) were measured. Vitamin E and/or omeprazole treated rats had significantly less XO% levels than H/R only group (0.36, 0.38 and 0.57, respectively). Similarly, the MDA levels were significantly lower in vitamin E and/or omeprazole received rats than H/R only rats (88.8, 97.9 and 122.6, respectively). All rats treated with omeprazole and/or vitamin E had better biochemical and histopathological levels compared to H/R rats (p < 0.05). Histopathological results show that Group 5 (H/R only) had significantly more intestinal damage when compared with Group 6 (vitamin E + H/R), Group 7 (omeprazole + R/H) and Group 8 (vitamin E + omeprazole + H/R) (p < 0.001). Grade 2 and 3 intestinal damages were only in Group 5 and there were no statistical difference between in Groups 6, 7 and 8 (p > 0.001). Omeprazole and/or vitamin E may protect the biochemical and histopathological intestinal damage of H/R injury in rats. These drugs may be beneficial in the prophylaxis of NEC in humans as well. PMID- 18427814 TI - The surgical management of fistula-in-ano in a specialist colorectal unit. AB - INTRODUCTION: Fistula-in-ano can be associated with a number of conditions, including Crohn's disease. The majority, however, are classified as idiopathic or cryptoglandular. The aim of this study was to review the outcome of surgical management of fistula-in-ano in a specialist colorectal unit. MATERIALS AND METHODS: One hundred and four consecutive patients underwent surgery for anal fistulae between 1st January 2000 and December 2004. Data was analysed in two main groups, according to the aetiology, cryptoglandular (n = 86) and Crohn's disease (n = 18). Follow-up data was available on 91 patients. RESULTS: In the cryptoglandular group, 62 patients had an inter-sphincteric tract, of which 48 underwent a single-stage fistulotomy. Of those patients with a trans-sphincteric tract, six patients underwent a single-stage fistulotomy, 13 had a seton and staged fistulotomy. Follow-up data revealed that two fistulae recurred. The median number of procedures in this group was 1 (range 1-3). There was a significant difference in the inpatient stay depending of Park's classification (p = 0.001). In the Crohn's group, three patients with an inter-sphincteric tract underwent a single-stage fistulotomy, two patients with a trans-sphincteric tract had single-stage fistulotomy, and five required a loose seton and staged fistulotomy. Eight patients had multiple fistulae which required long-term setons. Four patients from this group eventually required proctectomy. In the Crohn's group, there was a significantly increased complexity of surgery and higher recurrence. This was reflected in an increased inpatient length of stay and a greater reliance on imaging (p = 0.001). The median number of procedures in this group was 3 (range 1-5). DISCUSSION: The majority of cryptoglandular fistula in-ano were treated by primary fistulotomy or staged fistulotomy with a loose seton. This was associated with a low recurrence rate and low rates of faecal incontinence. There was a low reliance on imaging techniques in this group. However, we would urge caution when dealing with fistula-in-ano related to Crohn's disease. In this group of patients, the fistulae tended to be more complex and require additional imaging and multiple procedures. PMID- 18427815 TI - Astrocytic tau pathology positively correlates with neurofibrillary tangle density in progressive supranuclear palsy. AB - Tufted astrocytes (TAs) are considered reliable, specific markers for the neuropathologic diagnosis of progressive supranuclear palsy (PSP). It is known that neurofibrillary tangles (NFTs) may relate directly to neurodegeneration, but the role of glial tau pathology is not well determined. To examine the hypothesis that TAs are as pathogenic as NFTs and that both might have a common accumulation, we evaluated the topographic relationship between TAs and NFTs in 12 cases of PSP. The sections of 13 different parts of the brain were stained using the Gallyas-Braak method, and TAs and NFTs were counted and compared statistically. The number of TAs significantly correlated with that of NFTs in the central gray matter, pontine nuclei, and tegmentum, which are responsible for the main symptoms in PSP. In the examined allocortex, however, NFTs were abundant without accompanying TAs. Staining with the specific antibody for 4-repeat tau (RD4) and 3-repeat tau (RD3) was performed to clarify this discrepancy from the standpoint of tau isoforms. NFTs in the entorhinal cortex were stained with both RD3 and RD4, but NFTs in the premotor cortex were stained with only RD4. The nature of NFTs in the allocortical area was different from that of the isocortex in PSP. TAs in the isocortex may share the same pathologic cascade with NFTs stained only by RD4. These results suggest that TAs are part of the same pathologic process as NFTs in PSP. PMID- 18427816 TI - Copy number alterations of the polycomb gene BMI1 in gliomas. AB - Gliomas are heterogeneous tumours that grow in an uninhibited fashion, and these brain tumour cells share numerous characteristics with neural stem cells. The BMI1 gene encodes a component of the polycomb protein complex regulating epigenetically gene activity via histone modification. It functions for instance during the development of the central nervous system and maturation of neural cells. BMI-1 protein expression is deregulated in several forms of cancer and gene amplification has been identified in mantle cell lymphomas. Since BMI1 is located at chromosome 10p, a region implicated frequently in brain tumourigenesis, we investigated the genetic status and the corresponding expression patterns of BMI1 in a series of 100 low- and high-grade primary and recurrent gliomas. Chromogenic in situ hybridisation (CISH) with probes directed against BMI1 at 10p13 and the centromere of chromosome 10 was used in the analyses. Of all gliomas, 59% demonstrated aberrant copy numbers of BMI1. Deletions of the BMI1 locus were found in most types of tumours, and in a univariate survival analysis these cases had poor prognosis. Increased copy numbers of the BMI1 locus (3-5 copies) were found in all histological types, especially in high-grade astrocytomas. No difference in prognosis between cases with normal copy numbers and cases with increased copy numbers could be observed. This data suggests that BMI1 gene is aberrant at the chromosomal level in a subset of gliomas, and possibly contributes to brain tumour pathogenesis. PMID- 18427817 TI - Myenteric neurons of the ileum that express somatostatin are a target of prion neuroinvasion in an alimentary model of sheep scrapie. AB - Neuroinvasion of the enteric nervous system by prions is an important step in dissemination to the brain, yet very little is known about the basic process of enteric neuroinvasion. Using an alimentary model of neonatal disease transmission, neuroinvasion by scrapie prions in the ileum of lambs was detected by immunohistochemical staining for the disease-associated form of the prion protein, PrPSc. Odds ratios (OR) were determined for the frequency of PrPSc staining within enteric somata categorized by plexus location (myenteric, submucosal) and neurochemical staining (PGP 9.5, neural nitric oxide synthase, somatostatin, substance P, and vasoactive intestinal polypeptide). PrPSc was observed in 4.48 +/- 4.26% of myenteric neurons and 2.57 +/- 1.82% of submucosal neurons in five lambs aged 208-226 days but not in a lamb aged 138 days. The relative frequency of PrPSc within enteric somata was interdependent on plexus location and neurochemical type. Interestingly, PrPSc was observed more frequently within myenteric neurons than in submucosal neurons (PGP 9.5; OR = 1.72, 95% confidence interval = 1.21-2.44), and was observed within the myenteric plexus approximately 4x (2.16-6.94) more frequently in somatostatin neurons than in the general neural population stained by PGP 9.5. Nerve fibers stained for somatostatin were present in the mucosa and near PrPSc staining within Peyer's patches. The results suggest that somatostatin-expressing enteric neurons, with fiber projections near Peyer's patches, but with somata present in greatest proportion within the myenteric plexus, are an early target for neuroinvasion by scrapie prions and could serve an important role in neural dissemination. PMID- 18427818 TI - Acetabular component orientation in intra- and postoperative positions in total hip arthroplasty. AB - INTRODUCTION: In total hip arthroplasty (THA), acetabular component orientation has critically important effects on dislocation, range of motion, polyethylene wear, pelvic osteolysis, and component migration. The differences in the pelvic orientation in the intraoperative lateral position for insertion of acetabular component during operation and that in the postoperative supine position for evaluation of acetabular component orientation will be one of the factors, which make outliers in acetabular component orientation. We compared acetabular component orientation between intraoperative lateral position and postoperative supine position in 100 consecutive primary THAs. MATERIALS AND METHODS: A total of 100 consecutive primary THAs (between October 2004 and December 2005) in 100 patients performed by a single surgical team were investigated. Intraoperative anteroposterior radiographs of pelvis in the lateral position and postoperative anteroposterior radiographs of pelvis in the supine position were taken. Acetabular component orientation (vertical tilt and anteversion) were measured using computer software. RESULTS: The absolute values of difference between measurements in the two positions were 5.3 degrees +/- 4.5 degrees (mean +/- SD) for vertical tilt and 5.1 degrees +/- 3.7 degrees for anteversion. The difference in the vertical tilt between the two positions was significant (P < 0.0001). CONCLUSION: The difference in the acetabular component orientation between the two positions, which might be caused by the difference between intra- and postoperative pelvic orientation, should be considered during THA. PMID- 18427819 TI - Low-dose X-irradiation promotes mineralization of fracture callus in a rat model. AB - OBJECTIVES: This study investigated the hypothesized beneficial effect of low dose irradiation (LDI) on fracture callus mineralization in a rat model. METHODS: Seventy-two male Sprague-Dawley rats were averagely randomized into LDI group (rats treated with LDI) and SHAM group (rats treated with sham irradiation). Right after either LDI or sham irradiation, a standardized closed fracture on the right femur was established. At 2, 3 and 4 weeks postfracture, 12 rats in each group were euthanized. Fracture callus was assessed by using radiography and MicroCT for callus bridging, peripheral quantitative computed tomography (pQCT) for quantifying bone mineral content (BMC) and cross sectional area (CSA), confocal laser scanning microscopy for measuring area fraction of fluorescence labeling (AFFL) and four-point bending test for examining mechanical properties. RESULTS: The CSA and AFFL were found to be 22 and 33% smaller in the LDI group compared to the SHAM group at 2 weeks (P<0.05 for both), whereas the BMC and AFFL were 15 and 34% higher in the LDI group at 3 weeks (P<0.05 for both). The changing patterns were consistent with the findings in 3-D MicroCT reconstructions. The mechanical parameters (Max-Load, Stiffness and Energy) were also 18, 30 and 24% higher in the LDI group than in the SHAM group at 3 weeks (P<0.05 for all). At 4 weeks, there was no difference found for all assessments between the two groups. CONCLUSION: The results indicated LDI promoted mineralization at the stage of hard callus formation in a rat fracture model. PMID- 18427820 TI - Changes in serum levels of receptor activator of nuclear factor-kappaB ligand, osteoprotegerin, IL-6 and TNF-alpha in patients with a concomitant head injury and fracture. AB - INTRODUCTION: Several reports indicated that interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF- alpha) play important regulatory roles in bone remodeling and homeostasis. In addition, receptor activator of nuclear factor kappaB ligand (RANKL) and osteoprotegerin (OPG) have been shown to be important regulators of osteoclastogenesis during bone remodeling, and their expressions were examined during fracture healing in a mouse model of tibial fracture. However, studies linking RANKL, OPG, IL-6 and TNF-alpha in patients with head injury and fracture are lacking. PATIENTS AND METHODS: Within the first few hours of admission to hospital and at 4, 8, and 12 weeks after the injury, we evaluated changes in serum levels of RANKL, OPG, IL-6 and TNF-alpha in 24 male patients with a concomitant head injury and fracture and in 26 male patients with fracture only. These levels were compared with those found in 36 healthy controls. RESULTS: The RANKL/OPG ratios were found to significantly lower in patients with a concomitant head injury and fracture than in the controls immediately after admission and at 4, 8, and 12 weeks after the injury. In addition, RANKL/OPG ratios were significantly lower in patients with a concomitant head injury and fracture than in those with fracture at 8 and 12 weeks after the injury. The serum IL-6 levels were significantly higher in patients with a concomitant head injury and fracture than in the controls upon admission, and at 4, 8, and 12 weeks after the injury. Moreover, the serum IL-6 levels were significantly higher in patients with a head injury and fracture than in those with just a fracture at 4, 8, and 12 weeks after the injury. CONCLUSIONS: Based on these changes in the profiles of RANKL, OPG, and IL-6 and the RANKL/OPG ratio, altered repair of a fracture can occur in patients with a concomitant head injury and fracture. PMID- 18427821 TI - A novel 22-bp deletion mutation in a Chinese family with X-linked hypohidrotic ectodermal dysplasia. AB - X-linked hypohidrotic ectodermal dysplasia (XLHED) is the most common form of the ectodermal dysplasias characterized by an abnormal development of eccrine sweat glands, hair and teeth. Pathogenic mutations in the ED1 gene have been identified. In this family, a 22-bp deletion mutation of exon 8 in the ED1 gene was found in the affected members but not in the healthy individuals and 100 unrelated controls. We add new variant to the knowledge of ED1 mutations in XLHED. PMID- 18427822 TI - Influence of substance-P on cultured sebocytes. AB - Acne is a complex, chronic and common skin disorder of pilosebaceous units. Although it is known that exacerbation of acne results from emotional stress, the nature of the association between stress and acne remains unclear. This is due in part to the lack of substantial evidence regarding the participation of cutaneous neurogenic factors in the pathogenesis of acne. Culture of sebocytes provides a new insight into the participation of neuropeptides, notably substance P (SP), in the pathophysiology of acne. To examine the possible involvement of neurogenic factors in the pathogenesis of acne, we used immunohistochemistry and RT-PCR to compare the expression of interleukin-1 (IL-1), interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-alpha), peroxisome proliferators activated receptors gamma (PPAR-gamma) on the cultured sebocytes stimulated by SP. IL-1 is primarily proinflammatory cytokines to stimulate the expression of genes associated with inflammation. IL-6 is a pleiotropic cytokine with a wide range of biological activities and regulates inflammation. TNF-alpha is a pleiotropic pro inflammatory cytokine that exerts multiple biologic effects. PPAR-gamma is a nuclear hormone receptor and plays a unique role in stimulating sebocyte lipogenesis. More numerous immunoreactivity to IL-1, IL-6, TNF-alpha and PPAR gamma and increased RNA amplification for IL-1, IL-6, TNF-alpha and PPAR-gamma were observed after addition of SP compared with the control. This study reveals that SP is involved in the pathogenesis of acne. PMID- 18427823 TI - Femur fracture during abdominal breech delivery. AB - BACKGROUND: While vaginal breech delivery, although rare, can cause femur fracture, abdominal breech delivery is not expected to cause it. CASE: A 2,490-g female infant was delivered at term by elective cesarean section for breech presentation. She sustained a fracture of the femur shaft. A simple splinting led to a complete healing of the fracture without sequelae. CONCLUSION: Although abdominal breech delivery reduces the risk of birth trauma, we must be aware that femur fracture can occur regardless of the mode of delivery. PMID- 18427824 TI - Lower genital tract rhabdomyosarcoma: case series and literature review. AB - OBJECTIVE: Rhabdomyosarcoma (RMS) of the lower genital tract is a rare tumor. It tends to occur in childhood in the vagina and in rare cases, RMS can originate in the uterine cervix, with a peak incidence in the 2nd decade. METHODS: A hospital based tumor registry was searched to find all patients with female genital tract RMS, which were treated between 1999 and 2004. The medical records of all patients were reviewed. A single pathologist reviewed all pathologic specimens. RESULTS: Among the 1,528 patients with genital tract malignancies, six RMS were found: three vaginas, three cervixes. All patients presented with vaginal bleeding. Mean age of patients was 16(13-30). The lesions in vagina were clinically staged as stage I (2), stage II (1) and in cervix were stage I (2) and stage III(1). All of the patients were treated with surgery and adjuvant chemotherapy. One patient with cervical RMS was treated with adjuvant chemo radiation. Two patients with cervical RMS died from the large size and extent of the disease 9 and 11 months after diagnosis, but all patients with vaginal RMS remain alive after a mean follow-up of 38 months. CONCLUSION: Most patients present with vaginal bleeding and a palpable cervical or vaginal mass. While the optimal management of these tumors is uncertain, primary therapy with wide local excision and chemotherapy can result in prolonged survival and cure in patients with early stage RMS. Vaginal lesions have a better prognosis than cervical lesions. In patients with un-embrional RMS, large size of lesion, cervical origin and extent of disease, survival rates was decreased. PMID- 18427825 TI - Angulation of the styloid process in Eagle's syndrome. AB - In a retrospective study, we investigated and compared the angulation and the length of the styloid process between patients operated for Eagle's syndrome and a control group by means of lateral skull and Towne's radiographs. Thirty patients with 51 symptomatic elongated styloid processes underwent surgery. As a control group, patients with chronic otitis media and trauma were included in the study and none of them had symptoms characteristic of an elongated styloid process. The length, medial and anterior angulation of the styloid processes of the patient and the control groups were measured on lateral skull and Towne's radiographs. The mean length of the styloid process was 5 cm on the right and 5.2 cm on the left in the patient group, whereas they were 2.8 and 2.6 cm, respectively, in the control group. The mean degree of anterior angulation in the patient group was 33.6 degrees on the right, 36.7 degrees on the left, whereas these were 21.4 degrees and 18.5 degrees , respectively, in the control group. There was a significant difference between the two groups for length and anterior angulation (P = 0.001). The mean medial angulation was 14 degrees on the right and 18.1 degrees on the left in the patient group, whereas these were 15 degrees and 16.3 degrees in the control group, respectively, and there were no significant differences between the two groups. The anterior angulation and the length of the styloid process are responsible for the symptoms of Eagle's syndrome. PMID- 18427826 TI - Role of the alpha v-integrin subunit in cell proliferation, apoptosis and tumor metastasis of laryngeal and hypopharyngeal squamous cell carcinomas: a clinical and in vitro investigation. AB - The objective of this study was to investigate the role of alpha v-integrin subunit (ITGAV, CD51) in invasion and metastasis of the laryngeal and hypopharyngeal squamous cell carcinomas and to evaluate whether an antisense oligonucleotide sequence (ASONs) targeting ITGAV gene can result in proliferative inhibition and induce to apoptosis of laryngeal carcinoma cell lines (Hep-2). Firstly, a tissue microarray contained 75 primary carcinomas, 29 non-cancerous normal tissues and 20 metastatic lymph nodes was constructed and used to detect the expression of ITGAV by immunohistochemistry. The changes of ITGAV expression from each group were assessed and correlated to the clinical parameters of the patients. Secondly, the ASONs targeting ITGAV gene was transfected into Hep-2 cells in vitro. The proliferative ability of the cells after transfection was measured by MTT assay and the apoptosis was detected using flow cytometry. Results showed that the expression of ITGAV was significantly correlated with differentiation and lymph node metastasis of these cancers. In vitro test showed that the proliferative ability of Hep-2 cells was significantly inhibited by ASONs in a way of concentration- and time-depending mode, and a significant apoptosis of Hep-2 cells was also observed after ASONs transfection. In conclusion, the expression of ITGAV was significantly correlated with differentiation and metastasis of the laryngeal and hypopharyngeal carcinomas; down-regulation of ITGAV gene could inhibit proliferation of Hep-2 cells and induce to its apoptosis. These results suggest that ITGAV gene may become a promising prognostic marker and new treatment target for these cancers. PMID- 18427827 TI - Objective measurement of nasal airway dimensions and resistance using acoustic rhinometry and rhinomanometry in habitual snorers compared with non-snorers. AB - Snorers represent a heterogeneous group that requires adequate assessment before recommending surgical treatment. Most studies of the pathophysiology of snoring and obstructive sleep apnea have emphasized anatomical abnormalities in the oropharyngeal and hypopharyngeal airways. It is still unclear if nasal airway restriction plays an important role in sleep-disordered breathing and there is no general consensus if treatment of nasal pathology should be included in the management of patients with snoring or sleep apnea. The aim of this study was to compare nasal dimensions and airflow resistance of habitual snorers with non snoring individuals by means of acoustic rhinometry and rhinomanometry. Sixty individuals were enrolled in this analytical cross-sectional study. They were divided in two groups: group A (case) consisted of 30 patients with a main complaint of chronic snoring referred to ear, nose, and throat (ENT) clinic of Hazrat-e-Rasoul University Hospital, Tehran, Iran. Group B (control) consisted of 30 individuals without any complaint of snoring. The subjects were assessed objectively with acoustic rhinometry and rhinomanometry. Nasal dimensions and airflow resistance were recorded for all individuals. The most common site of minimum cross-sectional area (MCA) was at the left concha-notch in both snoring and non-snoring individuals. Significant reduction of cross-sectional area of both isthmus and concha notches was seen in habitual snorers (P < 0.05). The mean total airflow resistances in both pressures of 75 and 150 Pa was higher in habitual snorers. Whereas, these differences were not statistically significant (P > 0.05). The results of our study illustrate that acoustic rhinometry, rhinomanometry may be helpful methods for quantitative assessments of nasal airway respiratory function, and configuration in snorers; especially to evaluate site of MCA, decreased nasal cross-sectional area and increased nasal airflow resistance in habitual snorers which may lead to OSA. PMID- 18427829 TI - Socioeconomic and occupational groups and Parkinson's disease: a nationwide study based on hospitalizations in Sweden. AB - OBJECTIVES: The aim of this study was to investigate whether there is an association between socioeconomic status, occupation and hospitalization for Parkinson's disease (PD). METHODS: A nationwide database was constructed by linking the Swedish Census to the Hospital Discharge Register to obtain data on all first adult hospitalizations for PD diagnosed in Sweden during the study period from 1987 to 2004. Standardized incidence ratios (SIRs) with 95% confidence intervals (CIs) were calculated. RESULTS: A total of 8,870 males and 3,724 females first hospitalizations for PD were retrieved during the study period for adults aged 30 and older. Significantly increased SIRs for hospitalization for PD were found for men with high socioeconomic status. Among male occupations, increased SIRs were noted for several occupational groups such as teachers, administrators and managers, farmers, sales agents, wood workers, and painters and wall paperhangers. For female occupations, an increased risk was observed only among assistant nurses. The significant SIRs varied between 1.08 and 1.60. CONCLUSIONS: The socioeconomic and occupational groups used in the present study have a relatively small effect on the population's likelihood of hospitalization for PD but could give a notion of future research on specific occupational exposures. PMID- 18427828 TI - Biogenesis of mRNPs: integrating different processes in the eukaryotic nucleus. AB - Transcription is a central function occurring in the nucleus of eukaryotic cells in coordination with other nuclear processes. During transcription, the nascent pre-mRNA associates with mRNA-binding proteins and undergoes a series of processing steps, resulting in export-competent mRNA ribonucleoprotein complexes (mRNPs) that are transported into the cytoplasm. Experimental evidence increasingly indicates that the different processing steps (5'-end capping, splicing, 3'-end cleavage) and mRNP export are connected to each other as well as to transcription, both functionally and physically. Here, we review the overall process of mRNP biogenesis with particular emphasis on the functional coupling of transcription with mRNP biogenesis and export and its relationship to nuclear organization. PMID- 18427830 TI - Cancer mortality in workers exposed to dieldrin and aldrin: over 50 years of follow up. AB - OBJECTIVE: Dieldrin and aldrin, pesticides widely used until the 1970s, have been under suspicion of being carcinogenic. In this study, overall and cause-specific mortality was assessed in a cohort of 570 employees occupationally exposed to the pesticides dieldrin and aldrin to investigate the long-term health effects, in particular carcinogenic effects. METHODS: All of the employees worked in the production plants between January 1954 and January 1970 and were followed for cause-specific mortality until 30 April 2006. Based on dieldrin levels in blood samples taken from 343 workers during the exposure period, the total intake of dieldrin was estimated for each individual subjects in the cohort. The estimated total intake ranged from 11 to 7,755 mg of dieldrin, with an average of 737 mg. RESULTS: Two hundred and twenty-six workers had died before 30 April 2006 compared with an expected number of 327.3, giving a standardized mortality ratio (SMR) of 69.0 (95% confidence interval (CI): 60.3-78.7). Overall cancer mortality was also significantly lower than expected (SMR: 76.4, 95% CI: 60.8-94.9). Also, none of the specific cancer sites showed a significant excess mortality and no association between exposure level and cancer mortality was found. CONCLUSION: The results from this study support findings from other epidemiological and recent animal studies concluding that dieldrin and aldrin are not likely human carcinogens. PMID- 18427831 TI - Levels of agreement for RR intervals and short-term heart rate variability obtained from the Polar S810 and an alternative system. AB - The aim of this study was to assess the agreement between HRV measures derived from a time series of RR intervals recorded by a standard 12-lead ECG (CP) and a commercially available RR interval recorder (S810). Thirty-three participants (19 males) (median age 36, range 20-63) underwent simultaneous, 5-min, supine RR interval recordings. Each RR interval time series was analysed using the software supplied with the recording equipment. Two comparisons were then made. First, a comparison of RR interval data recording and editing only was made. Second, comparisons were made for measures of HRV derived from edited RR interval data. Agreement between RR intervals and standard HRV measures were assessed using intraclass correlation coefficient and limits of agreement. Agreement of HRV measures derived from RR intervals recorded and edited by individual systems was not acceptable. Agreement analyses for the number of RR intervals recorded and edited by each systems software showed excellent intraclass correlation coefficients (ICC lower 95% CI > 0.75) and acceptably narrow limits of agreement (LoA). These data indicate that the number of RR intervals recorded by S810 can agree well those recorded from a standard 12-lead ECG. This is true even after application of system specific data editing procedures. Commercial RR-interval recorders may offer a simple, inexpensive alternative to full 12-lead ECG in the recording and editing of RR intervals for subsequent HRV analysis in healthy populations. PMID- 18427832 TI - Professor Victor Alekseevich Rogozkin. PMID- 18427834 TI - A genetically encoded ratiometric sensor to measure extracellular pH in microdomains bounded by basolateral membranes of epithelial cells. AB - Extracellular pH, especially in relatively inaccessible microdomains between cells, affects transport membrane protein activity and might have an intercellular signaling role. We have developed a genetically encoded extracellular pH sensor capable of detecting pH changes in basolateral spaces of epithelial cells. It consists of a chimerical membrane protein displaying concatenated enhanced variants of cyan fluorescence protein (ECFP) and yellow fluorescence protein (EYFP) at the external aspect of the cell surface. The construct, termed pHCECSensor01, was targeted to basolateral membranes of Madin Darby canine kidney (MDCK) cells by means of a sequence derived from the aquaporin AQP4. The fusion of pH-sensitive EYFP with pH-insensitive ECFP allows ratiometric pH measurements. The titration curve of pHCECSensor01 in vivo had a pK (a) value of 6.5 +/- 0.04. Only minor effects of extracellular chloride on pHCECSensor01 were observed around the physiological concentrations of this anion. In MDCK cells, the sensor was able to detect changes in pH secondary to H(+) efflux into the basolateral spaces elicited by an ammonium prepulse or lactate load. This genetically encoded sensor has the potential to serve as a noninvasive tool for monitoring changes in extracellular pH microdomains in epithelial and other tissues in vivo. PMID- 18427833 TI - Cell models for studying renal physiology. AB - The development over the past 20 years of a variety of cultured renal tubule cell lines derived from different parts of the renal tubule has provided invaluable powerful cell systems for in vitro analyses of the various tubule segment specific biochemical functions and ion transport processes. Immortalized cell lines have been established using different hybrid gene constructs, most of them carrying the immortalizing simian virus 40 large T antigen (Tag) gene. The development of transgenic mice carrying unregulated Tag, and of others in which the expression of Tag remains controlled, has made it possible to establish permanent cell lines derived from microdissected or immunoselected renal proximal, distal, and collecting duct tubules. This review summarizes the different strategies of cellular immortalization used and the most frequently used human, rabbit, rat, and mouse tubule cell lines. This review provides an overview of the use of immortalized mouse tubule cell lines for in vitro analyses of various tubule cell-specific functions and the regulation of ion transporters and membranous channels. The advantages of using primary cultures of isolated tubules dissected from physiopathological models of transgenic mice are also discussed. PMID- 18427835 TI - Identification of P2X(4) receptor transmembrane residues contributing to channel gating and interaction with ivermectin. AB - Ivermectin (IVM), a large macrocyclic lactone, specifically enhances P2X(4) receptor-channel function by interacting with residues of transmembrane (TM) helices in the open conformation state. In this paper, we used cysteine-scanning mutagenesis of rat P2X(4)-TMs to identify and map residues of potential importance for channel gating and interaction with IVM. The receptor function was unchanged by mutations in 29 different residues, and among them, the IVM effects were altered in Gln(36), Leu(40), Val(43), Val(47), Trp(50), Asn(338), Gly(342), Leu(346), Ala(349), and Ile(356) mutants. The substitution-sensitive Arg(33) and Cys(353) mutants could also be considered as IVM-sensitive hits. The pattern of these 12 residues was consistent with helical topology of both TMs, with every third or fourth amino acid affected by substitution. These predominantly hydrophobic-nonpolar residues are also present in the IVM-sensitive Schistosoma mansoni P2X subunit. They lie on the same side of their helices and could face lipids in the open conformation state and provide the binding pocket for IVM. In contrast, the IVM-independent hits Met(31), Tyr(42), Gly(45), Val(49), Gly(340), Leu(343), Ala(344), Gly(347), Thr(350), Asp(354), and Val(357) map on the opposite side of their helices, probably facing the pore of receptor or protein and playing important roles in gating. PMID- 18427836 TI - Fas/FasL-dependent apoptosis of hepatocytes induced in rat and patients with Clonorchis sinensis infection. AB - To investigate the injury and pathogenesis of Clonorchis sinensis (C. sinensis) to hepatocytes, the liver samples from Wistar rats and patients with C. sinensis infection were examined. The typical histopathological findings of clonorchiasis were observed in rats 4 to 12 weeks postinfection, and majority hepatocytes exhibited hydropic degeneration, even some hepatocytes showed densely condensed nuclei suggesting apoptosis in liver tissue. Apoptosis was found around the central vein or portal areas of liver tissue in rat infected with C. sinensis by transferase uridyl nick end labeling (TUNEL) assay. Compared with normal control, TUNEL-positive cells in liver tissue increase from 4 to 12 weeks postinfection with the peak at 8 weeks. Furthermore, the expression of mRNA and protein of Fas, FasL, and caspase-3 was stronger in infected group than normal control using semiquantitative reverse transcriptase polymerase chain reaction (RT-PCR) and immunohistochemistry. Autopsy specimens from four patients infected with C. sinensis have the same findings detected by histopathology, TUNEL, and immunohistochemistry. These findings suggest that the C. sinensis can stimulate both hepatocytic apoptosis and hydropic degeneration that may be responsible for relationship between clinical manifestations and liver lesions in patients with clonorchiasis. These data also indicate a role for Fas/FasL-mediated pathway in the apoptosis that occurs in response to C. sinensis infection. PMID- 18427837 TI - Differential regulation of cellular maturation in chondrocytes and osteoblasts by glycine. AB - Previous studies have demonstrated the functional expression, by osteoblasts, of N-methyl-D-aspartate (NMDA) receptors responsible for the promotion of cellular differentiation in bone. We have now evaluated the possible role of the endogenous co-agonist of NMDA receptors, glycine (Gly), in chondrogenesis. In ex vivo organotypic cultures of fetal mouse tibias, proximal and distal cartilaginous primordia were significantly increased in the presence of Gly, with the osteogenic center being unchanged. Exposure to Gly drastically increased mRNA expression of the calcified chondrocyte marker osteopontin, without markedly affecting that of a proliferating chondrocyte marker or a hypertrophic chondrocyte marker, as shown in organotypic cultures by in situ hybridization analysis. Gly significantly increased Ca2+ accumulation, osteopontin mRNA expression, and alkaline phosphatase activity in cultured rat costal chondrocytes, without significantly affecting those in cultured rat calvarial osteoblasts. The increase induced by Gly was significantly prevented by an NMDA receptor channel blocker and an antagonist at the Gly site on NMDA receptors, but not by an inhibitory Gly receptor antagonist or a Gly transporter inhibitor, in cultured chondrocytes. Constitutive mRNA expression was seen for NR1, NR2D, and NR3A subunits of NMDA receptors, but not for Gly receptors and transporters, in cultured chondrocytes. Corresponding immunoreactive proteins were detected for NR1 and NR2D subunits in cartilaginous zones of fetal mouse tibias. Thus, Gly might, at least in part, play a role as a trophic factor in the mechanisms associated with chondral calcification through the Gly site of NMDA receptors functionally expressed by chondrocytes in rodent cartilage. PMID- 18427838 TI - Neuronal connections between central and enteric nervous system in the locust, Locusta migratoria. AB - The number and location of neurons, in the central nervous system, that project into the frontal connective was studied in the locust by using retrograde neurobiotin staining. Staining one frontal connective revealed some 70 neurons in the brain. Most of these were located within both tritocerebral lobes. Additional groups of neurons were located within the deutocerebrum and protocerebrum. Some 60 neurons were labelled in the suboesophageal ganglion. These formed nine discernable populations. In addition, two neurons were located in the prothoracic ganglion and two neurons in the first abdominal neuromere of the metathoracic ganglion. Thus, some 250 neurons located within the head ganglia, and even neurons in thoracic ganglia, project into the ganglia of the enteric nervous system. This indicates that the coordination between the central and enteric ganglia is much more complex than previously thought. With the exception of some previously described dorsal unpaired median neurons and a few motor neurons in the head ganglia, the identity and function of most of these neurons is as yet unknown. Possible functions of the neurons in the thoracic ganglia are discussed. PMID- 18427839 TI - Effects of evaporative cooling on the regulation of body water and milk production in crossbred Holstein cattle in a tropical environment. AB - The aim of this study was to determine how evaporative cooling modifies body function with respect to water metabolism and other variables relevant to milk synthesis in crossbred cattle. The study was conducted on two groups of 0.875HF:0.125RS crossbred Holstein cattle (87.5%) housed in an open-sided barn with a tiled roof (non-cooled animals) and in a close-sided barn under an evaporative cooling system (cooled animals). The maximum ambient temperature and relative humidity for the non-cooled group were 33 degrees C and 61%, with the corresponding values for the evaporatively cooled barn being 28 degrees C and 84%, respectively. The temperature humidity index (THI) of under non-cooled conditions was higher (P < 0.05) than that in the cooled barn. Rectal temperatures and respiration rates of non-cooled animals were higher (P < 0.05) than those of cooled animals. Daily dry matter intake (DMI) of cooled animals was higher while water intakes were lower (P < 0.05) than those of non-cooled animals. The mean absolute values of plasma volume, blood volume, and extracellular fluid (ECF) of cooled animals were significantly higher (P < 0.05) than those of non-cooled animals throughout all stages of lactation. Milk yields of cooled animals were higher by 42%, 36% and 79% on average than those of non cooled animals during early-, mid- and late-lactation, respectively. The decline in milk yields as lactation advances was markedly apparent in late-lactating non cooled animals, while no significant changes in milk composition at different stages of lactation were observed in either group. Mean arterial plasma concentrations, arteriovenous concentration differences (A-V differences) and the extraction ratio across the mammary gland for acetate, glucose and triglyceride of cooled animals were not significantly different compared with values for non cooled animals. No differences were seen in plasma hormonal levels for triiodotyronine (T(3)) and insulin-like growth factor-1 (IGF-1), but plasma cortisol and thyroxine (T(4)) levels tended to be lower in non-cooled animals. This study suggests that low cooling temperature accompanied by high humidity influences a galactopoietic effect, in part through increases in ECF, blood volume and plasma volume in association with an increase in DMI, which partitions the distribution of nutrients to the mammary gland for milk synthesis. Cooled animals were unable to maintain high milk yield as lactation advances even though a high level of body fluids was maintained during long-term cooled exposure. The decline in milk yield, coinciding with a decrease in net energy for lactation as lactation advances, could be attributed to a local change within the mammary gland. PMID- 18427840 TI - Independent evaluation of a clinical prediction rule for spinal manipulative therapy: a randomised controlled trial. AB - A clinical prediction rule to identify patients most likely to respond to spinal manipulation has been published and widely cited but requires further testing for external validity. We performed a pre-planned secondary analysis of a randomised controlled trial investigating the efficacy of spinal manipulative therapy in 239 patients presenting to general practice clinics for acute, non-specific, low back pain. Patients were randomised to receive spinal manipulative therapy or placebo 2 to 3 times per week for up to 4 weeks. All patients received general practitioner care (advice and paracetamol). Outcomes were pain and disability measured at 1, 2, 4 and 12 weeks. Status on the clinical prediction rule was measured at baseline. The clinical prediction rule performed no better than chance in identifying patients with acute, non-specific low back pain most likely to respond to spinal manipulative therapy (pain P = 0.805, disability P = 0.600). At 1-week follow-up, the mean difference in effect of spinal manipulative therapy compared to placebo in patients who were rule positive rather than rule negative was 0.3 points less on a 10-point pain scale (95% CI -0.8 to 1.4). The clinical prediction rule proposed by Childs et al. did not generalise to patients presenting to primary care with acute low back pain who received a course of spinal manipulative therapy. PMID- 18427841 TI - Neither the WAD-classification nor the Quebec Task Force follow-up regimen seems to be important for the outcome after a whiplash injury. A prospective study on 186 consecutive patients. AB - A classification of injury and a follow-up schedule were proposed by the Quebec Task Force (QTF) in 1995. No general agreement about the clinical usefulness of the WAD-classification or of the suggested follow-up regimen exists. A series of 186 consecutive cases seen in the emergency room during the acute phase after a whiplash injury was prospectively studied for 1 year. All findings including history and physical findings were recorded using standardized QTF protocols. In one group follow-up visits were done according to the QTF regimen: at 1, 3, 6, 12 weeks and 1 year after the accident; in a control group no visit was scheduled. The outcome variable was neck pain at 1 year after the accident. After 1 year, 18% of the total number of patients had significant neck pain. Risk factors for chronic neck pain at 1 year after whiplash injury were: neck pain before the accident and a high degree of emotional distress at the time of the accident; both factors independently associated with a tenfold increased risk of developing chronic neck pain. Neither the WAD classification nor the QTF follow-up regimen could be linked to a better outcome. In this study the outcome was associated with patient-specific characteristics and not with physical signs of injury, the depth of the initial evaluation or the follow-up regimen. PMID- 18427842 TI - Does scoliosis causes low bone mass? A comparative study between siblings. AB - The aim of this study is to assess the prevalence low bone mass among girls with adolescent idiopathic scoliosis (AIS) and their siblings. The subjects of this study were Saudi Arabian girls with AIS. Patients had their weight and height measured to calculate their body mass index (BMI). Clinical examination and investigations were done to rule out any other cause of scoliosis. All had bone mineral density (BMD) measurement of hip area and the spine using DEXA scan, Hologic Inc. Patients with a BMD of < -2.6 was taken as osteoporotic and those between < -1 and -2.5 was taken as osteopenic for analysis. As control subjects, siblings of the patients with normal spine had their BMI calculated and BMD measurement done. We were able to analyze the data of 32 girls with an average age of 18.42 +/- 5.71 (14-26) years with mean BMI of 17.7 +/- 0.69 (16.5-18.5) kg/M2. Analysis of the scans of the hip revealed that 62.5% of the patients were osteoporotic with BMD of 0.837 (0.697-0.936) +/- 0.04, T-score -3.8 +/- 0.56 ( 2.6 to -3.9) and Z-score. Nine (28.1%) were osteopenic with BMD of 0.768 +/- 0.15 (0.638-0.878), mean T-score of -1.6 (-1.1 to 2.5) and Z-score -3.5 +/- 0.63 (-2.9 to -3.9). Analysis of BMD of the spine showed similar results. In comparison to the scoliotics, girls with normal spine had higher BMI and BMD which was statistically significant at P < 0.001. T- and Z-score was also lower in scoliotic girls in comparison with girls with normal spine significant at P < 0.001 (CI 95%). Our study indicates that the scoliosis causes osteopenia and osteoporosis among girls while their siblings with normal spine remain with normal bone mass. PMID- 18427844 TI - Optimization of cutting schemes for the evaluation of molecular electrostatic potentials in proteins via Moving-Domain QM/MM. AB - This work presents new developments of the moving-domain QM/MM (MoD-QM/MM) method for modeling protein electrostatic potentials. The underlying goal of the method is to map the electronic density of a specific protein configuration into a point charge distribution. Important modifications of the general strategy of the MoD QM/MM method involve new partitioning and fitting schemes and the incorporation of dynamic effects via a single-step free energy perturbation approach (FEP). Selection of moderately sized QM domains partitioned between C (alpha) and C (from C=O), with incorporation of delocalization of electrons over neighboring domains, results in a marked improvement of the calculated molecular electrostatic potential (MEP). More importantly, we show that the evaluation of the electrostatic potential can be carried out on a dynamic framework by evaluating the free energy difference between a non-polarized MEP and a polarized MEP. A simplified form of the potassium ion channel protein Gramicidin-A from Bacillus brevis is used as the model system for the calculation of MEP. PMID- 18427843 TI - Clinimetric evaluation of active range of motion measures in patients with non specific neck pain: a systematic review. AB - The study is to provide a critical analysis of the research literature on clinimetric properties of instruments that can be used in daily practice to measure active cervical range of motion (ACROM) in patients with non-specific neck pain. A computerized literature search was performed in Medline, Cinahl and Embase from 1982 to January 2007. Two reviewers independently assessed the clinimetric properties of identified instruments using a criteria list. The search identified a total of 33 studies, investigating three different types of measurement instruments to determine ACROM. These instruments were: (1) different types of goniometers/inclinometers, (2) visual estimation, and (3) tape measurements. Intra- and inter-observer reliability was demonstrated for the cervical range of motion instrument (CROM), Cybex electronic digital instrument (EDI-320) and a single inclinometer. The presence of agreement was assessed for the EDI-320 and a single inclinometer. The CROM received a positive rating for construct validity. When clinical acceptability is taken into account both the CROM and the single inclinometer can be considered appropriate instruments for measuring the active range of motion in patients with non-specific neck pain in daily practice. Reliability is the aspect most frequently evaluated. Agreement, validity and responsiveness are documented less frequently. PMID- 18427845 TI - Intestinal obstruction: trends in imaging utilization and their influence in its rising hospital bill. AB - The purpose of the study was to evaluate trends in the utilization of different imaging modalities and review how imaging utilization practices affect hospital charges for patients with intestinal obstruction. All patients discharged with a primary diagnosis of intestinal obstruction during 6 fiscal years (1999-2004) were retrospectively studied. We obtained data on patients' demographics, procedures, outcomes, imaging services utilization, and hospital and imaging charges from our institution's transition system (a clinical and financial decision support software system). The institutional review board approved this study. Surgery was performed in 26% of patients in 1999 and in 40% in 2004 (p = 0.01) with the mortality rate significantly (p < 0.01) dropping from 3.8% to 0.4%. A total of 5,292 abdominal imaging studies were obtained; 93% of those were either abdominal radiographs or abdomino-pelvic computed tomography (CT) scans. CT studies per patient increased from 0.5 in 1999 to 1 in 2004 (p < 0.01), while abdominal radiographs (mean = 2.4) did not significantly change over the entire study period (p = 0.6). Average imaging charges doubled during the study period ($1,572 to $3,012, p < 0.01). Average hospital charges increased from $18,138 in 1999 to $32,808 in 2004 (p < 0.01). The fraction of hospital charges attributed to imaging varied between 8.7% and 9.2%. CT utilization for intestinal obstruction increased from 1999 to 2004 without modality substitution. While hospital and imaging charges have significantly increased, the fraction represented by imaging has remained constant, suggesting that imaging is an unlikely cause for the increase in hospital charges. PMID- 18427846 TI - Aortic injury with Chance fracture in a child. AB - To present the findings in a 10-year-old male with a Chance fracture and associated aortic vascular injury and review of the literature on the subject. We reviewed the clinical and imaging findings in a patient who was in a serious motor vehicle accident, sustaining a Chance fracture of the lumbar spine and associated aortic and common iliac vessel injury. We also accomplished a literature review. Evidence of an associated aortic injury was available on the initial computed tomography (CT) study, but the findings were more exquisitely demonstrated on reconstructed studies of the lumbar spine. Our literature review indicated that aortic injury associated with Chance fracture is relatively rare and often overlooked. Abdominal aortic injury although rare, can occur with Chance fractures. With the now present sophisticated CT imaging in most Emergency Rooms it is possible to obtain exquisite reconstructed images of all organ systems including the vascular tree. The images may provide enough details so that further delineation of any given problem may not be necessary. PMID- 18427847 TI - Bioconversion of starch processing waste to Phellinus linteus mycelium in solid state cultivation. AB - The objective of the experiment was to use starch processing waste as an alternative growth medium for cultivation of mycelia of the mushroom Phellinus linteus and to find an optimum condition under solid-state cultivation. Response surface analysis along with a central composite design was successfully applied to approximate the simultaneous effects of the substrate concentration (16-36 g l(-1)), pH (4.5-6.5), and temperature (25-35 degrees C) on the mycelial growth rate. In the model, pH and temperature significantly affected the mycelial growth but substrate concentration did not. The optimal substrate concentration, pH, and temperature for maximizing growth rate of P. linteus mycelia were found to be 16.5 g l(-1), pH 6.0, and 29.7 degrees C, respectively. Subsequent verification of these levels agreed with model predictions and the maximum mycelial growth rate at these conditions was 6.1 +/- 0.8 mm day(-1). Therefore, the results of the experiments suggest that starch processing waste could be utilized as a growth substrate for the cultivation of the mushroom mycelia of P. linteus, enhancing the usefulness of this byproduct of the starch manufacturing industry. This approach is likely to be useful for establishing similar parameters for the cultivation of other fungi. PMID- 18427848 TI - Degradation of raw feather by a novel high molecular weight extracellular protease from newly isolated Bacillus cereus DCUW. AB - Biotreatment of feather wastes and utilization of the degraded products in feed and foodstuffs has been a challenge. In the present study, we have demonstrated the degradation of feather waste by Bacillus cereus DCUW strain isolated during a functional screening based microbial diversity study on East Calcutta Wetland Area. A high molecular weight keratinolytic protease from feather degrading DCUW strain was purified and characterized. Moreover, utilization of degraded products during feather hydrolysis was developed and demonstrated. The purified keratinolytic protease was found to show pH and temperature optima of 8.5 and 50 degrees C, respectively. PMSF was found to inhibit the enzyme completely. The purified enzyme showed molecular weight of 80 kDa (from SDS-PAGE). The protease was found to have broad range substrate specificities that include keratin, casein, collagen, fibrin, BAPNA and gelatin. The protease was identified as minor extracellular protease (Vpr) by RT-PCR and northern blotting techniques. This is the first report describing the characterization of minor extracellular protease (Vpr) and its involvement in feather degradation in B. cereus group of organisms. PMID- 18427849 TI - Hydrogel coated monoliths for enzymatic hydrolysis of penicillin G. AB - The objective of this work was to develop a hydrogel-coated monolith for the entrapment of penicillin G acylase (E. coli, PGA). After screening of different hydrogels, chitosan was chosen as the carrier material for the preparation of monolithic biocatalysts. This protocol leads to active immobilized biocatalysts for the enzymatic hydrolysis of penicillin G (PenG). The monolithic biocatalyst was tested in a monolith loop reactor (MLR) and compared with conventional reactor systems using free PGA, and a commercially available immobilized PGA. The optimal immobilization protocol was found to be 5 g l(-1) PGA, 1% chitosan, 1.1% glutaraldehyde and pH 7. Final PGA loading on glass plates was 29 mg ml(-1) gel. For 400 cpsi monoliths, the final PGA loading on functionalized monoliths was 36 mg ml(-1) gel. The observed volumetric reaction rate in the MLR was 0.79 mol s( 1) m(-3) (monolith). Apart from an initial drop in activity due to wash out of PGA at higher ionic strength, no decrease in activity was observed after five subsequent activity test runs. The storage stability of the biocatalysts is at least a month without loss of activity. Although the monolithic biocatalyst as used in the MLR is still outperformed by the current industrial catalyst (immobilized preparation of PGA, 4.5 mol s(-1) m(-3) (catalyst)), the rate per gel volume is slightly higher for monolithic catalysts. Good activity and improved mechanical strength make the monolithic bioreactor an interesting alternative that deserves further investigation for this application. Although moderate internal diffusion limitations have been observed inside the gel beads and in the gel layer on the monolith channel, this is not the main reason for the large differences in reactor performance that were observed. The pH drop over the reactor as a result of the chosen method for pH control results in a decreased performance of both the MLR and the packed bed reactor compared to the batch system. A different reactor configuration including an optimal pH profile is required to increase the reactor performance. The monolithic stirrer reactor would be an interesting alternative to improve the performance of the monolith PGA combination. PMID- 18427850 TI - Measurement of functional microcirculatory geometry and velocity distributions using automated image analysis. AB - This study describes a new method for analyzing microcirculatory videos. It introduces algorithms for quantitative assessment of vessel length, diameter, the functional microcirculatory density distribution and red blood-cell (RBC) velocity in individual vessels as well as its distribution. The technique was validated and compared to commercial software. The method was applied to the sublingual microcirculation in a healthy volunteer and in a patient during cardiac surgery. Analysis time was reduced from hours to minutes compared to previous methods requiring manual vessel identification. Vessel diameter was detected with high accuracy (>80%, d > 3 pixels). Capillary length was estimated within 5 pixels accuracy. Velocity estimation was very accurate (>95%) in the range [2.5, 1,000] pixels/s. RBC velocity was reduced by 70% during the first 10 s of cardiac luxation. The present method has been shown to be fast and accurate and provides increased insight into the functional properties of the microcirculation. PMID- 18427851 TI - Constraining the general linear model for sensible hemodynamic response function waveforms. AB - We propose a method to do constrained parameter estimation and inference from neuroimaging data using general linear model (GLM). Constrained approach precludes unrealistic hemodynamic response function (HRF) estimates to appear at the outcome of the GLM analysis. The permissible ranges of waveform parameters were determined from the study of a repertoire of plausible waveforms. These parameter intervals played the role of prior distributions in the subsequent Bayesian analysis of the GLM, and Gibbs sampling was used to derive posterior distributions. The method was applied to artificial null data and near infrared spectroscopy (NIRS) data. The results show that constraining the GLM eliminates unrealistic HRF waveforms and decreases false activations, without affecting the inference for "realistic" activations, which satisfy the constraints. PMID- 18427852 TI - A hybrid algorithm for solving the EEG inverse problem from spatio-temporal EEG data. AB - Epilepsy is a neurological disorder caused by intense electrical activity in the brain. The electrical activity, which can be modelled through the superposition of several electrical dipoles, can be determined in a non-invasive way by analysing the electro-encephalogram. This source localization requires the solution of an inverse problem. Locally convergent optimization algorithms may be trapped in local solutions and when using global optimization techniques, the computational effort can become expensive. Fast recovery of the electrical sources becomes difficult that way. Therefore, there is a need to solve the inverse problem in an accurate and fast way. This paper performs the localization of multiple dipoles using a global-local hybrid algorithm. Global convergence is guaranteed by using space mapping techniques and independent component analysis in a computationally efficient way. The accuracy is locally obtained by using the Recursively Applied and Projected-MUltiple Signal Classification (RAP-MUSIC) algorithm. When using this hybrid algorithm, a four times faster solution is obtained. PMID- 18427854 TI - Consistency principle in biological dynamical systems. AB - We propose a principle of consistency between different hierarchical levels of biological systems. Given a consistency between molecule replication and cell reproduction, universal statistical laws on cellular chemical abundances are derived and confirmed experimentally. They include a power law distribution of gene expressions, a lognormal distribution of cellular chemical abundances over cells, and embedding of the power law into the network connectivity distribution. Second, given a consistency between genotype and phenotype, a general relationship between phenotype fluctuations by genetic variation and isogenic phenotypic fluctuation by developmental noise is derived. Third, we discuss the chaos mechanism for stem cell differentiation with autonomous regulation, resulting from a consistency between cell reproduction and growth of the cell ensemble. PMID- 18427853 TI - A model for the neuronal substrate of dead reckoning and memory in arthropods: a comparative computational and behavioral study. AB - Returning to the point of departure after exploring the environment is a key capability for most animals. In the absence of landmarks, this task will be solved by integrating direction and distance traveled over time. This is referred to as path integration or dead reckoning. An important question is how the nervous systems of navigating animals such as the 1 mm(3) brain of ants can integrate local information in order to make global decision. In this article we propose a neurobiologically plausible system of storing and retrieving direction and distance information. The path memory of our model builds on the well established concept of population codes, moreover our system does not rely on trigonometric functions or other complex non-linear operations such as multiplication, but only uses biologically plausible operations such as integration and thresholding. We test our model in two paradigms; in the first paradigm the system receives input from a simulated compass, in the second paradigm, the model is tested against behavioral data recorded from 17 ants. We were able to show that our path memory system was able to reliably encode and compute the angle of the vector pointing to the start location, and that the system stores the total length of the trajectory in a dependable way. From the structure and behavior of our model, we derive testable predictions both at the level of observable behavior as well as on the anatomy and physiology of its underlying neuronal substrate. PMID- 18427856 TI - EURRECA: EURopean micronutrient RECommendations Aligned. Preparing the way: a European Commission Network of Excellence. PMID- 18427855 TI - Soy intake is related to a lower body mass index in adult women. AB - BACKGROUND: Experimental and epidemiologic studies suggest that soy may promote weight loss. AIM OF THE STUDY: The goal of this study was to examine the relation of soy intake with body weight over the lifespan of women with Caucasian, Japanese, and Native Hawaiian ancestry. METHODS: We assessed the relation between lifetime soy consumption and body mass index (BMI) among 1,418 women in Hawaii. All subjects reported anthropometric measures, regular diet, and soy intake throughout life. The lifetime soy questionnaire was completed again by a subset of 356 women 5 years after study entry and the kappa values indicated moderate agreement. We regressed soy intake on BMI at study entry and at age 21 while controlling for confounding variables, computed least square means, and performed trend tests. RESULTS: Higher soy consumption in adulthood was related to a lower BMI (P=0.02). This association was only significant for Caucasian women and for postmenopausal subjects. The women in the highest category also experienced a smaller annual weight change since age 21 (by 0.05 kg/year) than the low soy intake group (P=0.02). We observed no association between early life soy intake and BMI. High vegetable consumption was significantly associated with a higher soy intake among Caucasian women. CONCLUSIONS: In this study, women consuming more soy during adulthood had a lower BMI, but the relation was primarily observed for Caucasian and postmenopausal subjects. This indicates that the association may be due to other nutritional factors and behaviors common in women with high soy intake. PMID- 18427857 TI - How we will produce the evidence-based EURRECA toolkit to support nutrition and food policy. AB - BACKGROUND: There is considerable variation in the recommended micronutrient intakes used by countries within Europe, partly due to different methodologies and concepts used to determine requirements and different approaches used to express the recommendations. As populations become more mobile and multi national, and more traditional foods become available internationally, harmonised recommendations based on up to date science are needed. This was recognised by the European Commission's (EC) Directorate-General (DG) Research in their 2005 call for proposals for a Network of Excellence (NoE) on 'nutrient status and requirements of specific vulnerable population groups'. EURopean micronutrient RECommendations Aligned (EURRECA), which has 34 partners representing 17 European countries, started on its 5-year EC-funded programme in January 2007. The programme of work was developed over 2 years prior to submitting an application to the EC. The Network's first Integrating Meeting (IM) held in Lisbon in April 2007, and subsequent consultations, has allowed further refinement of the programme. AIM: This paper presents the rationale for the EURRECA Network's roadmap, which starts by establishing the status quo for devising micronutrient recommendations. The Network has the opportunity to identify previous barriers and then explore 'evidence-based' solutions that have not been available before to the traditional panels of experts. The network aims to produce the EURRECA 'toolkit' to help address and, in some cases, overcome these barriers so that it can be used by those developing recommendations. RESULTS: The status quo has been largely determined by two recent initiatives; the Dietary Reference Intake (DRI) reports from the USA and Canada and suggestions for approaches to international harmonisation of nutrient-based dietary standards from the United Nations University (UNU). In Europe, the European Food Safety Authority (EFSA) has been asked by the EC's Directorate-General for Health and Consumer Protection to produce values for micronutrient recommendations. Therefore, EURRECA will draw on the uniqueness of its consortium to produce the sustainable EURRECA toolkit, which will help make such a task more effective and efficient. Part of this uniqueness is the involvement in EURRECA of small and medium-sized enterprises (SMEs), consumer organisations, nutrition societies and other stakeholders as well as many scientific experts. The EURRECA toolkit will contain harmonised best practice guidance for a more robust science base for setting micronutrient recommendations. Hence, in the future, the evidence base for deriving nutrient recommendations will have greater breadth and depth and will be more transparent. CONCLUSIONS: The EURRECA Network will contribute to the broader field of food and nutrition policy by encouraging and enabling the alignment of nutrient recommendations. It will do this through the development of a scientific toolkit by its partners and other stakeholders across Europe. This will facilitate and improve the formulation of micronutrient recommendations, based on transparently evaluated and quantified scientific evidence. The Network aims to be sustainable beyond its EC funding period. PMID- 18427858 TI - Current micronutrient recommendations in Europe: towards understanding their differences and similarities. AB - BACKGROUND: Nowadays most countries in Europe have established their own nutrient recommendations to assess the adequacy of dietary intakes and to plan desirable dietary intakes. As yet there is no standard approach for deriving nutrient recommendations, they may vary from country to country. This results in different national recommendations causing confusion for policy-makers, health professionals, industry, and consumers within Europe. EURRECA (EURopean micronutrient RECommendations Aligned) is a network of excellence funded by the European Commission (EC), and established to identify and address the problem of differences between countries in micronutrient recommendations. The objective of this paper is to give an overview of the available micronutrient recommendations in Europe, and to provide information on their origin, concepts and definitions. Furthermore this paper aims to illustrate the diversity in European recommendations on vitamin A and vitamin D, and to explore differences and commonalities in approaches that could possibly explain variations observed. METHODS: A questionnaire was developed to get information on the process of establishing micronutrient recommendations. These questionnaires were sent to key informants in the field of micronutrient recommendations to cover all European countries/regions. Also the latest reports on nutrient recommendations in Europe were collected. Standardisation procedures were defined to enable comparison of the recommendations. Recommendations for vitamin A and vitamin D were compared per sex at the ages 3, 9 months and 5, 10, 15, 25, 50 and 70 years. Information extracted from the questionnaires and reports was compared focusing on: (1) The concept of recommendation (recommended daily allowance (RDA), adequate intake (AI) or acceptable range), (2) The year of publication of the report (proxy for available evidence), (3) Population groups defined, (4) Other methodological issues such as selected criteria of adequacy, the type of evidence used, and assumptions made. RESULTS: Twenty-two countries, the World Health Organization (WHO)/the Food and Agriculture Organization of the United Nations (FAO) and the EC have their own reports on nutrient recommendations. Thirteen countries based their micronutrient recommendations on those from other countries or organisations. Five countries, WHO/FAO and the EC defined their own recommendations. The DACH-countries (Germany, Austria and Switzerland) as well as the Nordic countries (Norway, Sweden, Finland, Denmark and Iceland) cooperated in setting recommendations. Greece and Portugal use the EC and the WHO/FAO recommendations, respectively and Slovenia adopted the recommendations from the DACH-countries. Rather than by concepts, definitions, and defined population groups, variability appears to emerge from differences in criteria for adequacy, assumptions made and type of evidence used to establish micronutrient recommendations. DISCUSSION: The large variation in current micronutrient recommendations for population groups as illustrated for vitamin A and vitamin D strengthens the need for guidance on setting evidence based, up-to-date European recommendations. Differences in endpoints, type of evidence used to set recommendations, experts' opinions and assumptions are all likely to contribute to the identified variation. So far, background information was not sufficient transparent to disentangle the relative contribution of these different aspects. CONCLUSION: EURRECA has an excellent opportunity to develop tools to improve transparency on the approaches used in setting micronutrient recommendations, including the selection of criteria for adequacy, weighing of evidence, and interpretation of data. PMID- 18427859 TI - Children with mixed language disorder do not discriminate accurately facial identity when expressions change. AB - We investigated the recognition of pairs of faces (same or different facial identities and expressions) in two groups of 14 children aged 6-10 years, with either an expressive language disorder (ELD), or a mixed language disorder (MLD), and two groups of 14 matched healthy controls. When looking at their global performances, children with either expressive (ELD) or MLD have few differences from controls in either face or emotional recognition. At contrary, we found that children with MLD, but not those with ELD, take identical faces to be different if their expressions change. Since children with mixed language disorders are socially more impaired than children with ELD, we think that these features may partly underpin the social difficulties of these children. PMID- 18427860 TI - Child and adolescent psychiatry and family status: a nationwide register-based study. AB - BACKGROUND: Much attention has been given to parental separation as a possible risk factor for adverse child development; however, little information is available regarding the family status of children referred to psychiatric facilities. AIMS: To assess the association between psychiatric illness in childhood and family status compared to the background population. METHODS: Data was derived from a national register on children referred to psychiatric facilities in Denmark and a databank containing detailed statistical information on the Danish society. RESULTS: Regardless of age significantly fewer children with psychiatric illnesses lived with both biological parents as compared to the background population (51.1% vs 73.3%). There were no gender differences. Analyses of the specific diagnoses association with family status revealed only few significant differences. CONCLUSIONS: Psychiatrically ill children are at increased risk of not living with both biological parents independent of age of the child. PMID- 18427861 TI - A twin study investigating the genetic and environmental aetiologies of parent, teacher and child ratings of autistic-like traits and their overlap. AB - In the present study we investigated phenotypic agreement between informants (parent, teacher and child self-report) on ratings of autistic-like traits and compared the genetic and environmental aetiologies of the informants' ratings and of their covariance. Parents and teachers of >2,500 pairs from a community twin sample completed an abbreviated Childhood Asperger Syndrome Test (CAST). The twins also completed an adapted self-report version of the CAST. Structural equation model-fitting was carried out. Correlations between raters were significant but moderate (0.16-0.33). The magnitude of heritability estimates of autistic-like traits varied across raters, being highest for parent-rated autistic-like traits (82-87%) and more modest for child self-reported autistic like traits (36-47%). Genetic overlap was significant but moderate across all raters. These findings are discussed in relation to population screening for autism and future genetic research. PMID- 18427862 TI - Prevalence of DSM-IV disorders in Chinese adolescents and the effects of an impairment criterion: a pilot community study in Hong Kong. AB - PURPOSE: To provide preliminary prevalence estimates of common DSM-IV (Diagnostic and Statistical Manual of Mental Disorders-4th Edition) disorders in a sample of Hong Kong Chinese adolescents. METHODS: 541 Chinese adolescents were recruited from Grades 7, 8 and 9 of 28 mainstream high schools in Hong Kong (mean age=13.8 years; SD=1.2). The adolescents and their parents were separately administered the Youth and Parent versions of DISC-IV (Diagnostic Interview Schedule for Children-Version 4), respectively. RESULTS: Based upon both symptom and impairment criteria, as required by DSM-IV, the overall prevalence estimate of DSM-IV disorders in our sample of Chinese adolescents was 16.4%. Estimates for such individual disorders/diagnostic groupings as anxiety disorders, depressive disorders, attention deficit/hyperactivity disorder (ADHD), oppositional defiant disorder (ODD), conduct disorder (CD), and substance use disorders were 6.9, 1.3, 3.9, 6.8, 1.7, and 1.1%, respectively. These rates were largely compatible with those reported in previous studies with perhaps lower rates of generalized anxiety disorder (GAD), depressive disorders, CD, and substance use disorders, but a higher rate of ODD. The rate of ADHD was somewhat higher, but this might reflect the current DSM-IV diagnostic practice. The rate of anxiety disorders was not as high as predicted from some previous questionnaire surveys. The application of an impairment criterion had discernible impacts on prevalence estimates, greater on anxiety and substance use disorders, but smaller on depressive and disruptive behavior disorders. There was a lack of gender difference in rates of ODD and CD. DISCUSSION AND CONCLUSION: While the findings reported here are broadly compatible with those of other studies, there may be cross-cultural differences in rates of some individual disorders, e.g., GAD, depressive disorders, ODD, CD, and substance use disorders, as well as in gender difference regarding rates of ODD and CD. However, exact comparison between studies is confounded by methodological differences in sample characteristics, measures, and case definition. Standardization of methodology in epidemiological surveys should allow more precise identification of any within- or between culture variations in prevalence estimation. PMID- 18427863 TI - Psychosocial interventions for disruptive and aggressive behaviour in children and adolescents: a meta-analysis. AB - BACKGROUND: The effects of psychotherapy in reducing aggressive behaviours in children and adolescents using meta-analysis were estimated. METHOD: Sixty-five studies were included, covering 4,971 cases. Teacher reported change in aggression, change in social functioning, and changes in parental distress were calculated. RESULTS: The mean effect size (ES) of change in aggression in studies with untreated controls was 0.62 and in studies without untreated controls the ES was 0.95. In studies with or without untreated controls, the ESs in teacher reported aggression was 0.41 and 0.63, the ESs in changes in social functioning was 0.42 and 0.49, and the ESs in changes in parental distress was 0.39 and 0.47, respectively. CONCLUSION: Psychosocial treatments aimed at reducing aggressive behaviour have positive effects and additional treatment effects are moderate. In the moderator analysis, studies with untreated controls obtained significantly larger ESs if the sample size was small. Similarly, in studies without untreated controls, studies presenting diagnostic information, and studies with younger children resulted in significantly larger ESs, and studies applying behavioural interventions obtained significantly larger ESs as compared to studies applying family therapeutic interventions. There is still a need to further develop effective outpatient interventions for children being disruptive, and especially for adolescents. PMID- 18427865 TI - Narcissism, self-esteem, and conduct problems: evidence from a British community sample of 7-11 year olds. AB - The aim of the current report was to investigate the relationship between narcissism, self-esteem and conduct problems in a British community sample of pre adolescent and young adolescent children (n = 659; 7-11 year olds). We demonstrated that narcissism is associated with conduct problems, but no evidence for an interaction between low self-esteem and high narcissism in the prediction of conduct problems was found. Whilst low self-esteem was associated with teacher reported (but not parent-reported) conduct problems at the bivariate level of analyses, multi-variate analyses showed that self-esteem yielded no significant effects, neither independently, nor in interaction with narcissism for either parent- or teacher reported conduct problems. However, self-esteem was predictive of self-reported conduct problems at both the bivariate and multivariate level of analysis, possibly due to shared method variance. The findings suggest an important role for narcissism for conduct problems in children as young as seven years old. PMID- 18427864 TI - Executive dysfunction in Turkish children at high risk for schizophrenia. AB - OBJECTIVE: To explore different aspects of executive function (i.e. sequencing, set shifting and mental flexibility) in children who are at high risk for schizophrenia by comparing them with normal controls. METHOD: The high risk (HR) group consisted of 30 children whose parents were diagnosed as schizophrenia. As the control group (CG) 30 children, whose parents did not meet any DSM IV diagnostic criteria for any psychiatric disorder, participated. They were age and sex matched with the HR group. For the evaluation of different domains of cognitive functions Wechsler intelligence scale for children-revised (WISC-R), and a group of neuropsychological tests, including Trail Making A-B Tests, Color Form Test, and Progressive Figures Test were administered. Behavioral problems were assessed using Hacettepe Adjustment Scale. RESULTS: The subjects in the high risk group had significantly lower scores on Trail Making A-B, Color Form, Progressive Figures Tests, as well as subtests and scores of WISC-R (Information, Comprehension, Similarities, Picture Completion, Block Design, Object Assembly and Coding subtests, Verbal, Performance and Full Scale IQ scores). There is no significant difference between the two groups in the frequency and severity of behavioral problems. CONCLUSION: Children of parents with schizophrenia displayed significantly greater number of difficulties in several areas of executive function, such as sequencing, set shifting, and mental flexibility, when compared to their controls. PMID- 18427866 TI - Temperament features in adolescents with ego-syntonic or ego-dystonic obsessive compulsive symptoms. AB - The present study evaluated whether different patterns of temperament may predict a different threshold of acceptability of obsessive-compulsive (OC) symptoms in adolescents. OC symptomatology was detected with the Leyton Obsessional Inventory Child Version (LOI-CV) and temperament was assessed using the tridimensional personality questionnaire in 2,775 high-school students. According to the LOI-CV scores, the adolescents were classified as high interference (interfering, ego dystonic symptoms) (HI), supernormal (noninterfering, ego-syntonic symptoms) (Sn) and controls (C) HI were 119 (4.3%), Sn 85 (3.1%) and C 2,571 (92.6%). The best predictor of belonging to HI or Sn groups was the temperament configuration of high Harm Avoidance (HA) and high Persistence (P). The feature that mainly distinguishes the two symptomatic groups were Novelty Seeking (NS) levels. Our data suggest that people characterized by pessimistic worry in anticipation of future problems, passive avoidant behaviour, rapid fatigability (high HA) and irresoluteness, ambitiousness, perseverance, perfectionism, enduring feelings of frustration (high P) might develop OC symptoms. Whether OC symptoms become ego syntonic or ego-dystonic seems to mainly depend on NS levels: low NS might protect people (with the prevention of "exploratory and active behaviours" that may elicit loss of control on symptoms) from the development of interfering OC symptoms. PMID- 18427867 TI - Ten-year increase in service use in the Dutch population. AB - Since earlier research has indicated an increase in mental health problems among Dutch children, we investigated whether service use for mental health problems has also increased. Subsequently, we investigated whether a possible increase could be explained by child, family and socio-demographic characteristics that increase the likelihood of service use. We compared two population samples of 6- to 18-year-olds, one assessed in 1993 and one in 2003. Chi-square tests were conducted to examine differences between the proportions of service-users. We performed a logistic regression to test whether care-promoting factors accounted for the effect of year. Results showed that service use increased from 1993 to 2003, but not among children with high CBCL scores. Having serious problems, living in a family other than two biological parents, and having educational problems all increased the likelihood of service use and became more present in the Dutch population. These variables accounted for 49% reduction in the Odds Ratio of the effect of year. Although the proportion of children who used services increased from 1993 to 2003, still a large number of children experience an unmet need. The increase in the number of children from a family structure other than two biological parents or who have educational problems is a worrisome development in itself. PMID- 18427868 TI - Psychometric properties and diagnostic ability of the separation anxiety scale for children (SASC). AB - This research describes the psychometric properties of a new child self-report measure, the separation anxiety scale for children (SASC), which assesses the frequency of symptoms of separation anxiety in children from 8 to 11. The factor solution with a sample of 1,201 children from 8 to 11, isolated three factors, discomfort from separation, worry about separation, and calm at separation, tested by confirmatory factor analysis, which accounted for 32.80% of the total variance. Results indicated that the SASC has a high internal consistency (alpha = 0.83). The results revealed a high test-retest reliability of the instrument, and a high number of correctly diagnosed cases. The correlation of scores with other anxiety measures shows that the variable most associated with separation anxiety is trait-anxiety. PMID- 18427869 TI - Catatonia in autism: implications across the life span. AB - BACKGROUND: There is increasing evidence that catatonia is an important source of impairment in adolescents and adults with autism. AIM: Review of the evaluation, diagnosis, differential diagnosis, and treatment of catatonia in autism. METHOD: Presentation and discussion of a case-vignette spanning early childhood to adulthood. RESULTS: Autistic and catatonic symptoms overlap, yet catatonia is diagnosable in about one of seven adolescents and young adults with autism. Case reports suggest that benzodiazepines and electroconvulsive therapy are effective treatments in the acute and maintenance phase for people with autism who develop catatonia. CONCLUSIONS: Catatonia should be assessed in people with autism when there is an obvious and marked deterioration in movement, vocalizations, pattern of activities, self-care, and practical skills. Benzodiazepines and electroconvulsive therapy are favored options for acute and maintenance treatment in these cases. Further studies on the possible biological-genetic overlap between autism and catatonia would be helpful. PMID- 18427870 TI - Psychosocial conditions and suicidal behavior in pregnant teenagers : a case control study in Brazil. AB - OBJECTIVE: To compare the psychosocial profile and suicidal behavior of 110 pregnant teenagers (PT) with 110 non-pregnant teenagers (NPT). METHOD: Subjects were matched by age and residential district. The research instruments used were a structured clinical interview and a self-reporting questionnaire based on psychometric scales whose answers were subjected to univariate and multivariate logistic regression. RESULTS: Prevalence in the PT and the NPT groups were found to be as follows: attempted suicide (20.0 vs. 6.3%); depression (26.3 vs. 13.6%); anxiety (43.6 vs. 28.0%). Univariate analysis revealed the following significant associations with pregnancy: relocation in the previous 3 years (odds ratio (OR) = 6); years of schooling < or =7 (OR = 3.4); repeating a year in school (OR = 2.4); dropping out of school (OR = 5.2); death of a parent during childhood (OR = 2.9); use of alcohol/drugs in the family (OR = 2.5); previous attempted suicide (OR = 3.6); suicide by a relative (OR = 2.1); threats of physical/sexual abuse (OR = 3.5); depression (OR = 2.2); low level of social support (OR = 4.2); traumatic events (OR = 5.1) and psychosocial difficulties (OR = 4.4); prior use of tobacco and marijuana (OR = 4.0 and 4.8 respectively); weekly intake of alcohol over the previous 12 months (OR = 4.2). Multivariate analysis identified the following associations: relocation (OR = 6.4); prior use of tobacco (OR = 2.9); dropping out of school for a period in excess of 6 months (OR = 2.3); suicide by a social acquaintance (OR = 2.5). CONCLUSION: The PT case group exhibited a psychosocial profile whose characteristics clearly differentiate this group from the NPT control group. Preventive mental health care is needed to help PT because their behavioral pattern exposes them to high risk for suicide. PMID- 18427871 TI - Anomalous connection of the inferior vena cava to the left atrium: a surgical error in closing an atrial septal defect. PMID- 18427872 TI - Tumor necrosis factor-alpha and interleukin-10 gene promoter polymorphisms in Turkish rheumatoid arthritis patients. AB - Tumor necrosis factor and interleukin 10 have been implicated in the pathogenesis of rheumatoid arthritis (RA). Certain single-nucleotide polymorphisms (SNPs) within the promoter region of the IL-10 and TNF genes have been associated with altered levels of circulating IL10 and TNF. We aimed to explore the association of IL-10 and TNF-alpha polymorphisms in Turkish RA patients. We analyzed the association of TNF-alpha (-308G/A, -238G/A, -376G/A) and IL10 (-1082G/A, -819C/T, -592C/A) polymorphisms in 98 Turkish patients with rheumatoid arthritis and 122 healthy subjects using ARMS-PCR. The correlation of these findings with RF positivity and erosive disease in RA patients was also sought. A significant association was found between having RA and -1082 G allele (p=0.008; OR=1.44, 95% CI 1.11-1.86). There was no association between RA and -819C/T polymorphism. Significant differences were observed in IL10 GCC and ACC haplotypes distribution between RA and control subjects (p=0.006; OR=1.46, 95% CI 1.13-1.89 and p=0.011; OR=1.43, 95% CI 1.09-1.88, respectively). No statistically significant association was found between TNF-alpha 308G/A, -238G/A, -376G/A polymorphisms and RA. No significant association was found between RF positivity and erosive disease and TNF-alpha, IL10 gene polymorphisms. In addition, when combined genotypes were analyzed, no significant difference was found between RA patients and healthy controls. Our findings suggest that IL-10 1082 G/A polymorphism or GCC, ACC haplotypes may be associated with RA in Turkish patients. PMID- 18427873 TI - Photodynamic therapy in endodontic treatment of deciduous teeth. AB - The purpose of this study was to evaluate photodynamic therapy in deciduous teeth with necrotic pulp by means of fully quantifying viable bacteria, before and after instrumentation and after the use of photodynamic therapy. Radicular canal cultures were conducted (n = 10): the first one was performed right after access and location of the radicular canal; the second was performed after the conclusion of chemical-mechanical instrumentation, and the last one after photodynamic therapy. The photodynamic therapy was performed with 4 J/cm energy low-intensity diode together with toluidine blue. The results (log(10)) were submitted to a descriptive analysis and Wilcoxon test. The percentage of reduction was submitted to the Mann-Whitney test. The instrumentation resulted in a reduction of 82.59% of viable bacteria, and, after photodynamic therapy, the microbial reduction observed was 98.37% (P = 0.0126). Photodynamic therapy is recommended as adjunct therapy for microbial reduction in deciduous teeth with necrotic pulp. PMID- 18427874 TI - Primary hydatidosis of the gluteus muscles: report of three cases. AB - BACKGROUND: Hydatid disease caused by the tapeworm Echinococcus granulosus is a worldwide problem especially in sheep and cattle raising countries. AIMS: Muscle involvement is most commonly encountered as recurrence of previously treated disease or concurrently with primary lesions of the liver or lung. Furthermore, the rarity of muscle hydatidosis has unique implications in diagnosis and management. METHODS: We report on three cases of primary echinococcus of the gluteus muscles presented in our clinic during a 10-year period. RESULTS: We have administered albendazole for one cycle of 28 days in two of our patients based on the size and appearance of the cyst. All patients underwent total pericystectomy without cyst rupture. We have not found any recurrences after minimum follow up of 12 months. CONCLUSIONS: Muscle echinococciasis respond well to surgical intervention. Complete and intact removal of the cyst in primary muscular hydatidosis should be considered curative. PMID- 18427875 TI - TB or not TB?: a case of isolated testicular TB with scrotal involvement. AB - Despite the genitourinary tract being the most common site affected by extrapulmonary TB, isolated testicular TB remains a rare clinical entity. In patients with co-morbidities such as hepatic impairment, treatment proves a challenge, as first-line hepatotoxic pharmaceuticals are contraindicated. Here, we report a case of isolated testicular TB with scrotal involvement, on a background of hepatic dysfunction. PMID- 18427876 TI - Assessment of clinical and social need in patients attending an inner-city psychiatric service. PMID- 18427877 TI - Surgical debulking in a case of chronic lymphoedema. AB - Lymphoedema can present with an array of distressing symptoms, which can pose significant management dilemmas. First-line treatment involves established therapies including elevation, manual drainage and compression hosiery. However, failure of these medical strategies can occasionally necessitate surgical intervention to alleviate symptoms. This case highlights the role of surgical debulking in the management of an intractable case of lower limb lymphoedema with symptoms so severe that the patient requested amputation of her left lower extremity. This report also describes other surgical modalities in the management of this debilitating condition. PMID- 18427878 TI - Complementary and alternative medicine use in Irish paediatric patients. AB - BACKGROUND: Although very little scientific data exists on the efficacy and side effects of complementary and alternative medicines, their profile and availability is increasing. Use among Irish children is unknown. AIMS: To determine the nature and prevalence of complementary and alternative medicines (CAM) use in our paediatric population. METHODS: Parental questionnaires were distributed in 13 paediatric settings over a 4-month period. RESULTS: There were 57% of parents reported using CAM for their child. Use was significantly higher in the 2-4 years age group (34/105, 32%, P = 0.005). The commonest medicinal CAMs used were vitamins (88%), fish oils (27%) and Echinacea (26%). The commonest non medicinal CAMs used were homeopathy (16%) and craniosacral therapy (14%). Use varied between paediatric specialties, with the highest in neurological patients (23/25, 92%, P = 0.005). Only 13% of parents had informed their Paediatrician of their child's CAM use. CONCLUSIONS: More than half of the children surveyed had used some form of CAM, usually without their Paediatrician's knowledge. PMID- 18427879 TI - The challenge of diagnosing arrhythmogenic right ventricular cardiomyopathy in the young. AB - We report two cases of arrhythmogenic right ventricular cardiomyopathy (ARVC) in the pediatric age group. In both cases, the diagnosis was considered and pursued but would not be made utilizing Task Force Criteria. The diagnosis was made based on the morphology of a single beat during exercise testing. We illustrate the difficulty of diagnosing ARVC in the young even with a heightened index of suspicion. PMID- 18427880 TI - A statement on ethics from the HEART Group. PMID- 18427881 TI - PHACES association: a vasculocutaneous syndrome. AB - PHACES association is a spectrum of anomalies that might occur in infants with large facial hemangiomas. Most infants with PHACES association have segmental hemangiomas of the head or neck. Cardiac and cerebrovascular anomalies might be the most important association, as they carry a significant risk of complications. This article summarizes the dermatologic, cardiac, and cerebral vascular findings in a cohort of infants diagnosed with PHACES association. All had large segmental facial hemangiomas and aortic arch abnormalities. Four of the five were not suspected of having arch obstruction prior to imaging studies because of the aberrant origin of both subclavian arteries, and 4/5 required either interventional or surgical repair for arch obstruction. In contrast to classic aortic coarctation, the aortic anomalies found in the cohort had unusually complex and unpredictable anatomic involvement. Cerebral vascular anomalies were identified in 5/5, and 2/5 had neurologic complications secondary to abnormal cerebral vascular supply. It is important for care providers to recognize this association that presents with a cutaneous stigma, as it is associated with potentially lethal and often unrecognized vascular anomalies. Earlier recognition of the associated vascular pathologies might enable preemptive treatments before potentially devastating and irreversible sequelae. PMID- 18427882 TI - Effect of rainfall regime and slope on runoff in a gullied loess region on the Loess Plateau in China. AB - Runoff was measured from seven plots with different slopes nested in Tuanshangou catchment on the Loess Plateau to study effect of slopes on runoff in relation to rainfall regimes. Based on nine years of field observation and K-mean clusters, 84 rainfall events were grouped into three rainfall regimes. Rainfall regime A is the group of events with strong rainfall intensity, high frequency, and short duration. Rainfall regime C consists of events with low intensity, long duration, and infrequent occurrence. Rainfall regime B is the aggregation of events of medium intensity and medium duration, and less frequent occurrence. The following results were found: (1) Different from traditional studies, runoff coefficient neither decreased nor increased, but presented peak value on the slope surfaces; (2) For individual plot, runoff coefficients induced by rainfall regime A were the highest, and those induced by rainfall regime C were the lowest; Downslope, the runoff coefficients induced by three rainfall regimes presented the same changing trend, although the peak value induced by regime A occurred on a shorter slope length compared to those by regime B and C; (3) Scale effect on runoff induced by rainfall regime A was the least, and that induced by rainfall regime C was the largest. These results can be explained by the interactions of crusting, soil moisture content, slope length and gradient, and erosion units, etc., in the context of different rainfall regimes. PMID- 18427883 TI - The contribution of marsh zones to water quality in Dutch shallow lakes: a modeling study. AB - Many lakes have experienced a transition from a clear into a turbid state without macrophyte growth due to eutrophication. There are several measures by which nitrogen (N) and phosphorus (P) concentrations in the surface water can be reduced. We used the shallow lake model PCLake to evaluate the effects of three measures (reducing external nutrient loading, increasing relative marsh area, and increasing exchange rate between open water and marsh) on water quality improvement. Furthermore, the contribution of different retention processes was calculated. Settling and burial contributed more to nutrient retention than denitrification. The model runs for a typical shallow lake in The Netherlands showed that after increasing relative marsh area to 50%, total phosphorous (TP) concentration in the surface water was lower than the Maximum Admissible Risk (MAR, a Dutch government water quality standard) level, in contrast to total nitrogen (TN) concentration. The MAR levels could also be achieved by reducing N and P load. However, reduction of nutrient concentrations to MAR levels did not result in a clear lake state with submerged vegetation. Only a combination of a more drastic reduction of the present nutrient loading, in combination with a relatively large marsh cover (approximately 50%) would lead to such a clear state. We therefore concluded that littoral marsh areas can make a small but significant contribution to lake recovery. PMID- 18427884 TI - An approach toward understanding wildlife-vehicle collisions. AB - Among the most conspicuous environmental effects of roads are vehicle-related mortalities of wildlife. Research to understand the factors that contribute to wildlife-vehicle collisions can be partitioned into several major themes, including (i) characteristics associated with roadkill hot spots, (ii) identification of road-density thresholds that limit wildlife populations, and (iii) species-specific models of vehicle collision rates that incorporate information on roads (e.g., proximity, width, and traffic volume) and animal movements. We suggest that collision models offer substantial opportunities to understand the effects of roads on a diverse suite of species. We conducted simulations using collision models and information on Blanding's turtles (Emydoidea blandingii), bobcats (Lynx rufus), and moose (Alces alces), species endemic to the northeastern United States that are of particular concern relative to collisions with vehicles. Results revealed important species-specific differences, with traffic volume and rate of movement by candidate species having the greatest influence on collision rates. We recommend that future efforts to reduce wildlife-vehicle collisions be more proactive and suggest the following protocol. For species that pose hazards to drivers (e.g., ungulates), identify collision hot spots and implement suitable mitigation to redirect animal movements (e.g., underpasses, fencing, and habitat modification), reduce populations of problematic game species via hunting, or modify driver behavior (e.g., dynamic signage that warns drivers when animals are near roads). Next, identify those species that are likely to experience additive (as opposed to compensatory) mortality from vehicle collisions and rank them according to vulnerability to extirpation. Then combine information on the distribution of at risk species with information on existing road networks to identify areas where immediate actions are warranted. PMID- 18427885 TI - Volunteer macroinvertebrate monitoring: tensions among group goals, data quality, and outcomes. AB - Volunteer monitoring of natural resources is promoted for its ability to increase public awareness, to provide valuable knowledge, and to encourage policy change that promotes ecosystem health. We used the case of volunteer macroinvertebrate monitoring (VMM) in streams to investigate whether the quality of data collected is correlated with data use and organizers' perception of whether they have achieved these outcomes. We examined the relation between site and group characteristics, data quality, data use, and perceived outcomes (education, social capital, and policy change). We found that group size and the degree to which citizen groups perform tasks on their own (rather than aided by professionals) positively correlated with the quality of data collected. Group size and number of years monitoring positively influenced whether a group used their data. While one might expect that groups committed to collecting good quality data would be more likely to use it, there was no relation between data quality and data use, and no relation between data quality and perceived outcomes. More data use was, however, correlated with a group's feeling of connection to a network of engaged citizens and professionals. While VMM may hold promise for bringing citizens and scientists together to work on joint conservation agendas, our data illustrate that data quality does not correlate with a volunteer group's desire to use their data to promote regulatory change. Therefore, we encourage scientists and citizens alike to recognize this potential disconnect and strive to be explicit about the role of data in conservation efforts. PMID- 18427886 TI - What drives accelerated land cover change in central Argentina? Synergistic consequences of climatic, socioeconomic, and technological factors. AB - Synergistic combinations of climatic and land use changes have the potential to produce the most dramatic impacts on land cover. Although this is widely accepted, empirical examples, particularly involving deforestation in Latin America, are still very few. The geographic extent and causes of deforestation in subtropical seasonally dry forests of the world have received very little attention. This is especially true for the Chaco forests in South America, which are being lost at an alarming rate, sometimes higher than those reported for tropical forests. On this basis, the aims of this study were to analyze the changes in land cover that have occurred during the last three decades of the 20th century in the Chaco forests of central Argentina, and to explain the factors that have driven those changes. Results show major land cover changes. Approximately 80% of the area that was originally undisturbed forest is now occupied by crops, pastures, and secondary scrub. The main proximate cause of deforestation has been agricultural expansion, soybean cultivation in particular. This appears as the result of the synergistic convergence of climatic, technological, and socioeconomic factors, supporting the hypothesis of a multiple factor explanation for forest loss, while providing one of the very few existing analyses of changes in subtropical forests of the world. PMID- 18427887 TI - Solitary fibrous tumor of the pleura: an analysis of 13 cases. AB - BACKGROUND: Solitary fibrous tumor of the pleura is a rare soft-tissue tumor. In search of appropriate diagnosis and treatment methods, we present our experience with 13 patients. METHODS: The medical charts of 13 patients treated during the last 5 years were reviewed, as well as pathological records, including immunohistochemical stains. Follow-up data were obtained. In addition, a literature review with regard to treatment and clinical outcome was performed. RESULTS: Our series consisted of four men and nine women with a mean age of 47 years. Two were diagnosed before operation with ultrasonography-guided core needle biopsy. All patients underwent primary surgical treatment, and four of them were resected by video-assisted thoracic surgery. Seven tumors were malignant and the other six were benign. Immunohistochemical staining showed nestin was positive in three malignant solitary fibrous tumors of pleura (3/7), which were negative for CD34. Except for one, all patients were followed-up for 3 to 35 (mean, 14.5) months. Among them, one patient experienced a recurrence and one patient died of brain metastasis. CONCLUSION: Ultrasonography-guided core needle biopsy combined with immunohistochemical analysis might be a safe and rapid method to provide a confirmatory diagnosis before resection. For smaller, pedunculated tumors, video-assisted thoracic surgery could be a powerful and useful approach. We speculate that CD34-negative and nestin-positive might be a malignant marker for solitary fibrous tumor of pleura. PMID- 18427888 TI - Endoscopic antireflux surgery: are we there yet? PMID- 18427889 TI - Neurofuzzy is useful aid in diagnosing acute appendicitis. PMID- 18427891 TI - Trauma care in New Zealand: it's time to move ahead. AB - The quality of trauma care in New Zealand is good, but not optimal; it is characterized by skilled and dedicated staff relying on personal knowledge and institutional guidelines to deliver care. Quality assessment, standardization, and coordination of care continue to be hampered by the absence of a national trauma registry within the framework of a national trauma system. There is some hope that a planned regional pilot of the national trauma database will lead to more robust trauma system development, although this will only be achievable with solid governmental support. PMID- 18427892 TI - What World Health Assembly Resolution 60.22 means to those who care for the injured. AB - On May 23, 2007, the World Health Assembly (WHA) adopted WHA Resolution 60.22, "Health Systems: Emergency Care Systems," which called on the World Health Organization (WHO) and governments to adopt a variety of measures to strengthen trauma and emergency care services worldwide. This resolution constituted some of the highest level attention ever devoted to trauma care worldwide. This article reviews the background of this resolution and discusses how it can be of use to surgeons, emergency physicians, and others who care for the injured, especially in low- and middle-income countries. PMID- 18427893 TI - Percutaneous radiofrequency ablation of lung tumors in contact with the aorta: dangerous and difficult but efficient: a report of two cases. AB - Percutaneous imaging-guided tumor ablation is a widely accepted method for the treatment of primary and secondary lung tumors. Although it is generally feasible and effective for local tumor control, some conditions may affect its feasibility and effectiveness. Herein the authors report their experience with two patients with lung malignancies contiguous to the aorta who were successfully treated with radiofrequency ablation, even though it initially appeared highly risky due to the possible fatal complications. PMID- 18427894 TI - Transcatheter arterial chemoembolization for advanced hepatocellular carcinoma with inferior vena cava and right atrial tumors. AB - Advanced hepatocelluar carcinoma (HCC) with invasion of venous systems usually indicates not only a poor prognosis but also a contraindication for transcatheter arterial chemoembolization (TACE). This study evaluated the feasibility of TACE for advanced HCC with inferior vena cava (IVC) and right atrium (RA) tumors and, also, to search for the ideal embolization particle size. Twenty-six patients who had HCC invasion into the IVC included five patients with coexistent RA tumors that were treated with TACE. The chemoembolization method was cisplatin, doxorubicin, and mitomycin C mixed with Lipiodol and Ivalon. The selection of Ivalon particles was divided into two groups based on their size: (A) >180 microm, N = 9; and (B) 47-180 microm, N = 17. The overall response rate was 53.8% (14/26). Based on the response to TACE, the median survival period of the entire group was 4.2 months (range, 1.5 to 76.7 months). The median survival period of the 14 responders was 13.5 months (1.5-76.7 months), and that of the 12 nonresponders, 3.3 months (2.1 to 24.3 months) (p < 0.002). Comparing the two Ivalon particle sizes, the response rate was 12.5% (1/8 [corrected] patients) for group A and 72.2% [corrected] for group B (13/18 [corrected] patients) (p < 0.01). [corrected] No serious complication was observed post-chemoembolization. In conclusion, TACE is a safe and effective treatment for advanced HCC with IVC and RA tumors, and small Ivalon particles (47-180 microm) are superior to large ones (>180 microm). PMID- 18427895 TI - Homologous-restraint polymerase chain reaction: an efficient and rapid protocol to clone multiple homologous genes. AB - In this article, we present a novel protocol, called homologous-restraint polymerase chain reaction (HRPCR), for cloning multiple homologous genes. One of the homologous genes was cloned by consensus-degenerate hybrid oligonucleotide (CODEHOP) polymerase chain reaction (PCR) and sequenced. Primers of HRPCR were designed with 20 to 30 nt inverted to the known gene before the 5' end of the CODEHOP primers. The amplification of the known gene was restricted owing to the loop of the PCR product or the incorrect binding of the primers and the template. As a result, only unknown genes could be cloned. This protocol proved to be simple, rapid, and efficient. We applied this protocol to clone the multiple homologous genes of beta-1,4-N,6-O-diacetylmuramidase from the genomic DNA of Streptomyces griseus. PMID- 18427896 TI - Hypercoagulability: another potential mechanism of obstructive sleep apnea related cardiovascular disease? PMID- 18427898 TI - Anatomical variations in stylopharyngeus muscle insertions suggest interindividual and left/right differences in pharyngeal clearance function of elderly patients: a cadaveric study. AB - The stylopharyngeus plays a critical role in the clearance of the piriform recess. We dissected 78 sides of the pharynx from 55 donated cadavers and observed histology of another seven sides of the pharynx from seven cadavers. The stylopharyngeus consistently comprised (1) a descending muscle bundle surrounding the piriform recess and (2) an additional short sheet inserting into the tonsillar bed. Histologically, the former bundle connected to a thick fascia providing the lateral glossoepiglottic fold, extending along the submucosa of the piriform recess, and covering the thyroid cartilage, whereas the latter sheet intermingled with other pharyngeal wall muscles at and near the tonsillar bed. Notably, in 44.4% of female specimens, the additional sheet occupied a greater proportion in cross section than the descending muscle bundle. Given the different directions, the additional sheet seemed to check clearance function of the descending bundle for the piriform recess. Thus, particularly in women, interindividual differences in pharyngeal clearance were likely to depend on whether the additional sheet is strong or weak. Chin down in combination with tilting and rotating the head may represent effective exercises of the stylopharyngeus that could compensate for the disadvantages of additional insertion. PMID- 18427900 TI - Artifacts found during quality assurance testing of computed radiography and digital radiography detectors. AB - A series of artifact images, obtained over 5 years of performance testing, of both computed radiography (CR) and integrated digital radiographic X-ray imaging detectors are presented. The images presented are all either flat field or test object images and show artifacts previously either undescribed in the existing literature or meriting further comment. The artifacts described are caused by incorrect flat field corrections, a failing amplifier, damaged detector lines affecting their neighbors, lost information between neighboring detector tiles, image retention, delamination of a detector, poor setup of mechanical movements in CR, suckers damaging a CR plate, inappropriate use of grid suppression software, inappropriate use of a low pass spatial frequency filter, and unsharp masking filters. The causes and significance of the artifacts are explained and categorized as software or hardware related. Actions taken to correct the artifacts are described and explained. This work will help physicists, radiographers, and radiologists identify various image quality problems and shows that quality assurance is useful in identifying artifacts. PMID- 18427901 TI - Phytotoxic catechin leached by seeds of the tropical weed Sesbania virgata. AB - Sesbania virgata (Cav.) Pers (wand riverhemp) is a fast-growing tropical legume species that has been used for revegetation of riparian forests and rehabilitation of degraded areas and that exhibits an invasive behavior in certain regions of Brazil. Preliminary studies have shown that seed leachates inhibit the germination and development of seedlings of some crop species. In this study, we report that the seed leachates of S. virgata inhibit the growth of Arabidopsis thaliana and rice. The flavonoid (+)-catechin is found in high amounts in these leachates. It was active at concentrations of 50 microg ml(-1), and its effect was not distinguishable from the (+)-catechin obtained from a commercial source. We found that (+)-catechin is located in the seed coat and is rapidly released in high concentrations (235 microg per seed) at the beginning of imbibition. Quercetin was also detected in the seed coat of S. virgata, but it was not released from the seeds. Other phytotoxic compounds in the seed leachates were also detected. The fact that S. virgata releases high amounts of (+) catechin, which also has antimicrobial activity, and other phytotoxins from its seeds at the earliest stages of its development might represent some adaptative advantage to the seedling that contributes to its invasive behavior and successful establishment in different soils. PMID- 18427902 TI - The relationship between multimorbidity and patients' ratings of communication. AB - BACKGROUND: The growing interest in pay-for-performance and other quality improvement programs has generated concerns about potential performance measurement penalties for providers who care for more complex patients, such as patients with more chronic conditions. Few data are available on how multimorbidity affects common performance metrics. OBJECTIVE: To examine the relationship between multimorbidity and patients' ratings of communication, a common performance metric. DESIGN: Cross-sectional study SETTING: Nationally representative sample of U.S. residents PARTICIPANTS: A total of 15,709 noninstitutionalized adults living in the United States participated in a telephone interview. MEASUREMENTS: We used 2 different measures of multimorbidity: 1) "individual conditions" approach disregards similarities/concordance among chronic conditions and 2) "condition-groups" approach considers similarities/concordance among conditions. We used a composite measure of patients' ratings of patient-physician communication. RESULTS: A higher number of individual conditions is associated with lower ratings of communication, although the magnitude of the relationship is small (adjusted average communication scores: 0 conditions, 12.20; 1-2 conditions, 12.06; 3+ conditions, 11.90; scale range 5 = worst, 15 = best). This relationship remains statistically significant when concordant relationships among conditions are considered (0 condition groups 12.19; 1-2 condition groups 12.03; 3+ condition groups 11.94). CONCLUSIONS: In our nationally representative sample, patients with more chronic conditions gave their doctors modestly lower patient-doctor communication scores than their healthier counterparts. Accounting for concordance among conditions does not widen the difference in communication scores. Concerns about performance measurement penalty related to patient complexity cannot be entirely addressed by adjusting for multimorbidity. Future studies should focus on other aspects of clinical complexity (e.g., severity, specific combinations of conditions). PMID- 18427903 TI - Early relaparoscopy for management of suspected postoperative complications. AB - BACKGROUND: Diagnosis of complications after laparoscopic surgery is difficult and sometimes late. METHODS: We compared the outcome of patients who had early (<48 h) relaparoscopy for suspected postoperative complication to those where relaparoscopy was delayed (>48 h). RESULTS: During the study period, 7726 patients underwent laparoscopic surgery on our service. Of these, 57 (0.7%) patients had relaparoscopy for suspected complication. The primary operations were elective in 48 patients and emergent in nine. Thirty-seven patients had early, 20 had delayed, secondary operations. The most common indication in the early group was excessive pain (46%) followed by peritoneal signs in 35%. In the delayed group, the most common indication was signs of systemic inflammatory response syndrome in 30% and peritoneal signs in 25%. Relaparoscopy was negative in 16 (28%) patients with no difference between groups. The identified complication was treated laparoscopically in 37(65%) patients, and the rest were converted. The patients in the delayed group had a significantly longer hospital stay (p < 0.003) and had a higher rate of complications (p < 0.05). They also had a higher mortality rate (10% vs. 2.7%), but the difference was not statistically significant. CONCLUSIONS: A policy of early relaparoscopy in patients with suspected complications enables timely management of identified complications with expedient resolution. PMID- 18427904 TI - Prospective nonrandomized comparison between pylorus-preserving and subtotal stomach-preserving pancreaticoduodenectomy from the perspectives of DGE occurrence and postoperative digestive functions. AB - BACKGROUND: To determine the influence of pylorus preservation after pancreaticoduodenectomy, we compared the postoperative course of subtotal stomach preserving pancreaticoduodenectomy (SSPPD) and pylorus-preserving pancreaticoduodenectomy (PPPD). METHODS: A prospective, nonrandomized comparison of 77 consecutive patients undergoing PPPD (n = 37) or SSPPD (n = 40) between January 2003 and March 2007 was planned. The early postoperative course, dietary intake, and the incidence of delayed gastric emptying (DGE) were evaluated. RESULTS: SSPPD included significantly more cases of regional lymph node dissection (D2, PPPD 53% vs. SSPPD 80%) and portal vein resection. The median duration of surgery (457 vs. 520 min) was significantly shorter, and blood loss (619 vs. 1,235 ml) was significantly less in PPPD. Regarding postoperative clinical factors, the duration of nasogastric tube intubation (1 vs.1 day), days until solid diet (7 vs. 7 days), and the incidence of DGE (9% vs.10%) were similar in PPPD and SSPPD. However, the postoperative/preoperative body weight ratio (95% vs. 93%) was significantly higher, and the postoperative hospital stay (31 vs. 38 days) was significantly shorter in PPPD (p < 0.05). CONCLUSIONS: Despite the bias of the operative factors, the incidence of DGE and postoperative dietary intake after SSPPD was comparable with PPPD, and therefore, pylorus preservation seemed to have no impact on postoperative dietary intake or DGE. PMID- 18427905 TI - The use of a high-throughput luminescent method to assess CYP3A enzyme induction in cultured rat hepatocytes. AB - Assessment of a new chemical entity for cytochrome P450 (CYP) enzyme induction at an early stage in discovery is crucial to prevent potential drug-drug interactions. CYP3A, the most abundant CYP isoform in the liver, metabolizes approximately 50% of drugs currently on the market and is also a highly inducible enzyme. The use of both rat and human hepatocyte culture for the prediction of in vivo CYP3A induction has become refined and validated and is considered a standard in vitro model. The current evaluation of CYP3A enzyme induction involves the use of substrates requiring subsequent analysis of metabolites by high-performance liquid chromatography/mass spectrometry, which adds considerable time and cost. In the present study, we describe the use of a novel luminogenic substrate, luciferin-6'-pentafluoro-benzyl ether (PFBE), which allows for a fast and selective measurement of CYP3A enzyme induction in cultured rat hepatocytes. The extent of induction was evaluated using cells treated for 3 d with the prototypical inducers, dexamethasone, phenobarbital, and pregnenolone 16 alpha carbonitrile (PCN). Enzyme activity was measured in the treated cells either by the depentafluorobenzylation of luciferin-PFBE or the testosterone 6-beta hydroxylation. Using both methods, dexamethasone and PCN-treated cells exhibited strong CYP3A activity, whereas phenobarbital treatment resulted in a weak response. The fold induction varied between both methods, but this variability can be controlled by normalizing data from each treatment to a positive control. The results indicate that luciferin-PFBE is an attractive alternative to the use of conventional substrate, testosterone, providing a sensitive, robust, and rapid method compatible with the multiwell plate format for the assessment of CYP3A induction. PMID- 18427906 TI - 50 years ago in CORR: Postspinal anesthesia osteomyelitis of the lumbar spine P. L. Day MD and J. J. Hinchey MD CORR 1958;11:185-193. PMID- 18427907 TI - Incarcerated massive incisional hernia: extensive necrosis of the colon in a very obese patient. Surgical treatment and vacuum-assisted closure therapy: a case report. AB - We discuss a diabetic obese patient with an extensive necrosis of the ascending and transverse colon plus segmental necrosis of the small bowel incarcerated in a massive median incisional hernia below the umbilicus. After a blood test and an abdominal CT scan (without contrast dial), the patient underwent an urgent operation. We performed an extended right hemicolectomy, multiple segmental small bowel resections and a terminal ileostomy. The defect of the abdominal wall was treated with vacuum-assisted closure (VAC) therapy with good results. PMID- 18427908 TI - Severe degenerative change of multiple organs mediated by chronic active Epstein Barr virus infection with infected T-cell expansion. AB - We here report the case of a young Japanese woman diagnosed with chronic active Epstein-Barr virus (EBV) infection. Intensive therapy with the CHOP regimen was partially able to control virus expansion, but various central nervous system symptoms appeared and gradually progressed. EBV-encoded RNA, detected using in situ hybridization, disclosed the presence of EBV in liver and bone marrow tissue, and real-time PCR revealed the presence of EBV in the cerebrospinal fluid (CSF) and serum. CD3+CD4+CD8-CD56- T-cell expansion in the peripheral blood (PB) and CSF was also observed. Atrophic brain changes were progressive, and the patient died of central nervous system disturbance and pulmonary hemorrhage a year after diagnosis. Autopsy revealed that EBV-infected T lymphocytes with a phenotype similar to those seen in PB and CSF had infiltrated multiple organs: the lymph nodes, bone marrow, endocardium, pericardium, myocardium, spleen, liver, and spinal cord. There have been few previous reports of severe degenerative changes in the myocardium, liver, and spinal cord in patients with EBV infection. Although EBV occasionally infiltrates the central nervous system and brain, atrophic changes mediated by EBV are rare. The autopsy results of this case suggest the possibility of EBV-mediated, severe degenerative changes in multiple organs. PMID- 18427910 TI - Stability with unilateral external fixation in the tibia. AB - Unilateral external fixation can be used in the provisional or definitive treatment of tibial fractures. A properly applied fixator allows bony and soft tissue stability, whereas an improperly applied fixator achieves neither and can be a hindrance. The principles for the successful application of monolateral external fixation, including the rationale for choosing this type of device, the assembly of its components and deciding on planes of application, are discussed in this article. PMID- 18427911 TI - Correction of complex foot deformities using the V-osteotomy and the Ilizarov technique. AB - Complex foot deformity is a multiplanar deformity with or without foot shortening. It also includes deformed feet with poor soft-tissue coverage, relapsed or neglected cases, and those with acompanying problems such as leg length discrepancy, lower leg deformity, osteomyelitis and nonunions. Traditionally, correction of these deformities can be achieved by extensive soft tissue releases, osteotomies or arthrodesis with or without internal fixation. This usually involves excision of large appropriate bony wedges and has many disadvantages, including neurovascular injury, soft tissue problems and a shortened foot. We present our experience with a group of severe deformities of the foot that we managed using the V-osteotomy combined with the Ilizarov technique. We present our algorithm of management of complex foot and ankle deformities, together with our prerequisites for patient selection. A detailed description of the operative technique, postoperative care and possible complications is also presented. The combination of the Ilizarov technique and the V-osteotomy offers versatility in foot deformity correction, enabling correction of individual components of the deformity at rates that may be tailored to achieve accurate three-dimensional control. PMID- 18427909 TI - Biomechanics of the anterior cruciate ligament and implications for surgical reconstruction. AB - Injury to the anterior cruciate ligament (ACL) is regarded as critical to the physiological kinematics of the femoral-tibial joint, its disruption eventually causing long-term functional impairment. Both the initial trauma and the pathologic motion pattern of the injured knee may result in primary degenerative lesions of the secondary stabilisers of the knee, each of which are associated with the early onset of osteoarthritis. Consequently, there is a wide consensus that young and active patients may profit from reconstructing the ACL. Several factors have been identified as significantly influencing the biomechanical characteristics and the functional outcome of an ACL reconstructed knee joint. These factors are: (1) individual choice of autologous graft material using either patellar tendon-bone grafts or quadrupled hamstring tendon grafts, (2) anatomical bone tunnel placement within the footprints of the native ACL, (3) adequate substitute tension after cyclic graft preconditioning, and (4) graft fixation close to the joint line using biodegradable graft fixation materials that provide an initial fixation strength exceeding those loads commonly expected during rehabilitation. Under observance of these factors, the literature encourages mid-to long-term clinical and functional outcomes after ACL reconstruction. PMID- 18427912 TI - Open reduction and corrective ulnar osteotomy for missed radial head dislocations in children. AB - The treatment of chronic radial head dislocation remains controversial. Open reduction of the radial head in combination with correction of malalignment with ulnar osteotomy can be the key to a good surgical result. Between 2001 and 2006, 9 (6 female, 3 male, average age 8.4 (5-11) years) patients were treated surgically for chronic radial head dislocation by one surgeon. The time between trauma and surgery was 7 (1.5-14) months. The procedure consisted of open reduction of the dislocated radial head and reconstruction of the annular ligament in combination with an ulnar osteotomy. An upper arm cast was applied with the forearm in neutral rotation for six weeks. Plates were removed in all patients. Clinical and radiological evaluation took place preoperatively and after an average of 23 (10-49) months. At radiograph 8/9 showed a reduced radial head; in one an anterior subluxation was seen. The range of motion remained the same in 4 patients who had a full range of motion preoperatively. In 2/5 patients with loss of range of motion preoperatively, improvement was seen. There were no serious surgical complications beside one infection. Open reduction and corrective ulnar osteotomy shows good results for missed radial head dislocations in children. PMID- 18427913 TI - Internal femoral osteosynthesis after external fixation in multiple-trauma patients. AB - In this study the authors evaluate the results of internal synthesis of femoral fractures in polytraumatised patients initially treated by external fixation (EF). From January 2002 to December 2005, 39 femurs in 37 polytraumatised patients (average age 34.2 years, range 18-44) with closed fractures and an ISS>20 were initially treated with EF. There were three groups: Group A, 13 cases when conversion to internal osteosynthesis occurred after 4-7 days (average 5.6 days); Group B, 11 cases with a 4-6-month interval before internal osteosynthesis, and after investigation using MRI and scintigraphy with labelled leucocytes; Group C, the remaining cases treated definitively with EF. Time of healing, lower limb function, time of return to previous activities and short and long-term complications were evaluated at the follow-up. The average time of follow-up was 23 months. In Group A the time of bone healing was 123 days; there were no events of embolism but one case of pseudoarthrosis and one case of instrument failure. In Group B the time of bone healing was 274 days, with one case of pseudoarthrosis and one case of deep infection. In Group C the average healing time was 193 days, with 3 cases of screw (half-pin) osteolysis. Functional recovery was delayed by the presence of other fractures. EF is a simple, quick and safe procedure to stabilise fractures in polytraumatised patients. According to damage control orthopaedic (DCO) concepts, it is possible to replace EF with internal synthesis after an interval as this reduces the risks of internal osteosynthesis when performed in the emergency period. EF can also be maintained as definitive treatment but should a change to internal synthesis be needed, it is possible to do it safely after excluding bone infection. PMID- 18427914 TI - Minimally invasive ankle arthrodesis with a retrograde locking nail after failed fusion. AB - A retrograde nail with posterior-to-anterior (PA) locking into os calcis, talus and tibia was used to correct deformity and achieve fusion after failed fusion. A variety of methods have been published to achieve union of the ankle and subtalar joint in a failed fusion situation. We have studied a retrograde locking nail technique through a 2.5-cm incision in the non-weightbearing part of the sole of the foot. Remaining cartilage in the ankle joint, where necessary, was percutaneously removed through an anterior approach and the locking nail was inserted after reaming of os calcis, talus and tibia. Locking screw insertion was in the sagittal plane (p.a. direction), in talus os calcis and tibial diaphysis using a nail mounted jig. Ten patients were entered in the study (age 27-60 years). The initial aetiology for attempted fusion was post-traumatic in nine cases and rheumatic in one case. There were 25 previous operations in the cohort not leading to fusion. An additional temporary external fixator was used in four cases to reach and maintain the optimum position for the procedure. The intervention time was 30-75 min. Dynamisation of the nail was performed after four months under local anaesthesia. The mean duration of follow-up was 4 years (3-5.5 years). Radiologically and clinically, fusion was achieved in 16 weeks (range, 12-20 weeks). There was no loosening of the implant or implant failure. A leg length discrepancy was avoided using this technique. There was one complication with varus malunion in a heavy smoker which united after corrective osteotomy, revision nailing and bone grafting. Patient satisfaction was measured on a scale (not visual analogue) of 0 (not satisfied) to 10 (completely satisfied); overall satisfaction averaged 9.5 points (range, 6-10 points). The postoperative ankle-hindfoot score of the American Orthopedic Foot and Ankle Society averaged 73.5 points (range, 61-81 points). Retrograde locked nailing with locking in the sagittal plane is a reliable minimally invasive procedure to achieve fusion of the ankle and the subtalar joint after failed fusion. PMID- 18427915 TI - The "floating forearm" injury in a child: a case report. AB - The case of a eleven-year-old girl who had a fracture dislocation of the left elbow with entrapment of the ulnar nerve into the dislocated ulnar epicondyle anlage and unstable forearm fracture of the ipslateral upper extremity is described. This severe injury to the elbow and the ipsilateral forearm is termed "floating forearm" injury. The forearm was stabilized percutaneously and the elbow fracture dislocation, remaining unstable after internal fixation was treated with a pediatric elbow fixator with motion capacity. PMID- 18427916 TI - A life without pain: a case report. AB - Congenital insensitivity to pain is a rare condition with an abnormality of interpretation of painful stimuli. This case report illustrates how a sequence of injuries after no or trivial trauma incapacitated a young boy. Especially the bilateral collapse and dislocation of the hip is an unusual sequela of this disorder. PMID- 18427917 TI - Results of treatment of displaced supracondylar humeral fractures in children by percutaneous lateral cross-wiring technique. AB - Seventy children with displaced type II and III supracondylar fractures of the humerus were managed with percutaneous lateral cross-wiring technique from January 2006 to January 2007. There were 54 boys and 16 girls with a mean age of 6.1 +/- 3.07 years. All patients were operated within 24 h after trauma using the Dorgans percutaneous lateral cross-wiring technique. Patients were followed up for a mean period of 6.1 +/- 2.6 months and assessed both radiologically for union; and functionally and cosmetically according to Flynn's criteria. All patients achieved solid union. Functionally, all patients achieved satisfactory results, while cosmetically, 91.4% of patients had satisfactory results and 8.6% had unsatisfactory results. The most frequently occurring complications were minor pin tract infection in six patients, deep infection in two patients, and 32 patients suffered excessive granulation tissue formation mostly around the proximal pin. There was no iatrogenic neurological injury either for the ulnar or for the radial nerves. The obtained results and minor complications reported signify this technique as a viable treatment method for displaced type II and III supracondylar fractures in children. PMID- 18427918 TI - Percutaneous compression plating versus gamma nail for the treatment of pertrochanteric hip fractures. AB - The objective of this study is to compare percutaneous compression plating (PCCP) device with standard gamma nail (GN). A sample was prospectively followed and compared to a historical cohort: 82 intertrochanteric hip fractures in 81 patients treated with PCCP in 2004 versus 51 hip fractures treated with GN in 2003 (AO type 31A1, 31 A2). The main outcome measures were: surgery times, blood loss (Hb serum level and transfusions), complication, costs, for a 1-year follow up. The minimally invasive PCCP technique resulted in a lower blood loss and consequently lower transfusion need (statistically significant), fewer implant related complications and comparable surgery times. Overall surgical costs were lower for a comparable outcome in terms of healing and surgical time. PMID- 18427919 TI - The treatment of stable and unstable proximal femoral fractures with a new trochanteric nail: results of a multicentre study with the Veronail. AB - The incidence of hip fractures continues to rise. This study is the first evaluation of a new intramedullary implant, the Veronail, that provides double axis fixation into the femoral head and allows the surgeon to choose whether to use sliding or fixed locked proximal screw fixation for trochanteric femoral fractures. The fractures were classified according to the AO classification, and function was assessed with the Modified Harris Hip Score. 111 patients with trochanteric fractures were evaluated in eight Italian hospitals. The stable 31.A1 fractures were treated with sliding proximal screws, the subtrochanteric 31.A3 fractures with converging proximal screws, and the unstable 31.A2 fractures were treated with both types of proximal fixation. The unstable fractures treated with locked converging screws had the same function at one year as those treated with sliding screws. This study suggests a possible new method of treating unstable trochanteric femoral fractures. This may be the solution to prevent excessive collapse of the fracture with the resultant poor function and persisting pain noted in the literature. Two converging locked proximal screws seem to provide stable fixation in 31.A2 femoral fractures and produce as good a result as the use of traditional sliding screws. The role of converging locked proximal screws in unstable trochanteric fractures requires further evaluation. PMID- 18427920 TI - A pseudo-iatrogenic case of medial clavicular fracture. AB - Medial fractures are the least common type of clavicular fracture (2-10%). The patient is a 29-year-old gynaecology resident with hyper-laxity and sternoclavicular instability. The latter had been surgically stabilized with Dacron((R)) tape, which eroded the bone causing an usura. Acute right shoulder pain occurred 10 years later. CT revealed medial clavicular stress fracture. After 4 weeks of conservative management, internal fixation followed. Five months postoperatively the patient performed all activities without pain. In this patient the weakened medial clavicle due to usura clearly played a role in both the site and nature of the fracture. Furthermore, CT is essential in arriving at the correct diagnosis. PMID- 18427921 TI - High-pressure injection injury of the hand: an often underestimated trauma: case report with study of the literature. AB - The real extent of damage in high-pressure injection injuries (grease gun injuries, paint gun injuries, pressure gun in juries) is hidden behind a small and frequently painless punctiform skin lesion on the finger or the hand. These kinds of injuries require prompt surgical intervention with surgical debridement of all ischemic tissue. Possibility of a general intoxication by the fluid must always be ruled out. Postoperative intensive physiotherapy is essential for the final hand function. The initial benign aspect is frequently causing a delay for an adequate treatment while in the mean time the possibility for subcutaneous damage continuously increases. Because of this delay the chance of permanent reduced functionality in the hand or finger amputation raises. Not only the latency time to adequate treatment but also the injected fluid's nature, the pressure, the volume and the location of injection, has influence on the seriousness and extensiveness of subcutaneous damage. All these factors influence the functional outcome of the patient. PMID- 18427922 TI - External fixator-assisted acute shortening with internal fixation for leg length discrepancy after total hip replacement. AB - We report a case of a 51-year-old lady, who underwent a femoral shortening using a fixator assisted blade plate after total hip replacement. The patient had a total hip replacement on the other side with previous revisions, which resulted in a leg length discrepancy. We used the above technique to control the shortening and preserve the mechanical and anatomical axis of the femur. PMID- 18427923 TI - Traumatic anterior hip dislocation associated with anterior and inferior iliac spines avulsions and a capsular-labral lesion. AB - A 16-year-old man had a traumatic accident during motor-cross racing and sustained an anterior hip dislocation with fracture of anterior superior and anterior inferior iliac spines. The hip was emergently reduced and further imaging was obtained to evaluate the lesion and bony fragments. Computed tomography confirmed the presence of two large fragments and a small fragment from the antero-inferior acetabulum. The patient underwent open reduction and internal fixation of the iliac spines and the reparation of anterior hip capsule. No complications occur after 1 year of follow-up. We present the case and a review of the literature for this lesion. PMID- 18427924 TI - Fibular lengthening by Ilizarov method secondary to shortening by osteochondroma of distal tibia. AB - Osteochondroma is the most common benign bone tumour. They most commonly affect the long tubular bones and almost half of osteochondromata are found around the knee. Osteochondroma arising from the distal metaphysis of the tibia typically result in a valgus deformity of the ankle joint secondary to relative shortening of the fibula. This case describes the use of Ilizarov technique for fibular lengthening following excision of a distal tibial osteochondroma. A 12-year-old girl presented with a 3-year history of a large swelling affecting the lateral aspect of the right distal tibia. Plain radiographs confirmed a large sessile osteochondroma arising from the postero-lateral aspect of the distal tibia with deformity of the fibula and 15 mm of fibular shortening. The patient underwent excision through a postero-lateral approach and subsequent fibular lengthening by Ilizarov technique. The patient made excellent recovery with removal of frame after 21 weeks and had made a full recovery with normal ankle function by 6 months. The Ilizarov method is a commonly accepted method of performing distraction osteogenesis for limb inequalities; however, this is mainly for the tibia, femur and humerus. We are unaware of any previous cases using the Ilizarov method for fibular lengthening. This case demonstrates the success of the Ilizarov method in restoring both fibular length and normal ankle anatomy. PMID- 18427925 TI - Alteration of metal ions improves the activity and thermostability of aminoacylase from hyperthermophilic archaeon Pyrococcus horikoshii. AB - Recombinant L-aminoacylase (PhoACY) from a hyperthermophilic archeon, Pyrococcus horikoshii, is a zinc-containing metalloenzyme. When the zinc was substituted by Mn(2+) or Ni(2+), its specific activity was significantly increased with acetyl-L methionine as a substrate. The thermostability of PhoACY was improved when it was incubated with 1 mM Zn(2+), Mn(2+) or Ni(2+). The enzyme with external Zn(2+) addition had no significant loss of the activity when held at 90 degrees C for up to 12 h and moreover had more than a 10-fold longer half-life even at 100 degrees C, compared to the enzyme without Zn(2+) addition. A thermostable structure of the enzyme associated with zinc binding is described based on differential scanning calorimetry. PMID- 18427926 TI - Co-culture of arbuscular mycorrhiza-like fungi (Piriformospora indica and Sebacina vermifera) with plant cells of Linum album for enhanced production of podophyllotoxins: a first report. AB - Cell suspension cultures of Linum album were developed from internode portions of in vitro germinated plant in Gamborg's B5 medium supplemented with 0.4 mg naphthalene acetic acid/l. The highest biomass was 8.5 g/l with podophyllotoxin and 6-methoxypodophyllotoxin at 29 and 1.9 mg/l, respectively after 12 d cultivation. Co-cultures of L. album cells with axenically cultivable arbuscular mycorrhiza-like fungi, Piriformospora indica and Sebacina vermifera, were established for the first time. These enhanced podophyllotoxin and 6 methoxypodophyllotoxin production by about four- and eight-fold, respectively, along with a 20% increase in biomass compared to the control cultures. PMID- 18427927 TI - Esterification reactions catalyzed by lipases immobilized in organogels: effect of temperature and substrate diffusion. AB - Rhizomucor miehei lipase was immobilized in hydroxy(propylmethyl) cellulose or agar gels containing lecithin or AOT microemulsions. The effect of the diffusion of substrates and products to this catalyst was studied, as well as the effect of temperature on the initial rate of ester synthesis. The composition of the gel affects the reaction rate due to mass transport phenomena. The apparent activation energies were higher for the systems based on agar, independently of the microemulsion used, and lower for the systems based on AOT microemulsions, independently of the polymer used. PMID- 18427928 TI - Metabolic engineering of noviose: heterologous expression of novWUS and generation of a new hybrid antibiotic, noviosylated 10 deoxymethynolide/narbonolide, from Streptomyces venezuelae YJ003-OTBP1. AB - NovW, novU and novS genes have been characterized as dTDP-4-keto-6-deoxy-D glucose 3-epimerase, C-5 methyltransferase and dTDP-glucose 4-ketoreductase, respectively involved in noviose biosynthetic pathway. We have cloned and expressed the Streptomyces spheroids novWUS genes in S. venezuelae YJ003-OTBP1. This established the function of novWUS and, at the same time, it also proved that the noviosyl derivative of 10-deoxymethynolide(2)/narbonolide(4) obtained from S. venezuelae YJ003-OTBP1 is a novel hybrid antibiotic. PMID- 18427929 TI - Biotransformation of (+/-)-2-methylcyclohexanone by fungi. AB - The biotransformation of 2-methylcyclohexanone (1) using 16 fungal strains and some mushroom cultures was investigated. Fusarium sp. was one of the effective biocatalysts for oxidoreduction of 2-methylcyclohexanone (1). cis-2 Methylcyclohexanol (2a) was isomerized to trans-2-methylcyclohexanol (2b) by Fusarium sp. In addition, the corresponding lactones 3 was obtained by Baeyer Villiger oxidation using Fusarium sp. AP-2 (46%, 94% ee). PMID- 18427930 TI - Advances towards resonance assignments for uniformly--13C, 15N enriched bacteriorhodopsin at 18.8 T in purple membranes. AB - Solid state NMR spectra from uniformly (13)C, (15)N enriched bacteriorhodospin (bR) purified from H. salinarium were acquired at 18.8 T using magic angle spinning methods. Isolated resonances of 2D (13)C-(13)C spectra exhibited 0.50 0.55 ppm line-widths. Several amino acid types could be assigned, and at least 12 out of 15 Ile peaks could be resolved clearly and identified based on their characteristic chemical shifts and connectivities. This study confirms that high resolution solid state NMR spectra can be obtained for a 248 amino acid uniformly labeled membrane protein in its native membrane environment and indicates that site-specific assignments are likely to be feasible with heteronuclear multidimensional spectra. PMID- 18427931 TI - The Institute of Medicine's reports on quality and safety: paradoxes and tensions. PMID- 18427933 TI - Characterization of the lignin signature in Lake Mead, NV, sediment: comparison of on-line flash chemopyrolysis (600 degrees C) and off-line chemolysis (250 degrees C). AB - The distribution of lignin in sediment is a useful tool for tracing the transport of land-derived organic matter in an aquatic environment. Tetramethylammonium hydroxide (TMAH) flash chemopyrolysis, or chemolysis followed by GC-MS analysis can be used for evaluating the origin of organic carbon in sediments. TMAH chemopyrolysis or chemolysis of organic matter produces a myriad of semi-volatile products. Among these products are methylated phenols which are an indirect measure of lignin in sediment. In this study, total organic carbon, elemental carbon, and lignin were measured in Lake Mead sediments. This study indicates that terrestrial runoff makes a contribution to Lake Mead sediments, and that this contribution is most apparent in sediment that is close to the Las Vegas Wash. Two chemolysis methods (on-line and off-line) were examined and compared for detection of lignin phenols. The results from these sediment cores indicate that comparable results can be obtained from the two approaches, although detection levels are significantly lower for the off-line approach. PMID- 18427932 TI - A novel pepper membrane-located receptor-like protein gene CaMRP1 is required for disease susceptibility, methyl jasmonate insensitivity and salt tolerance. AB - Plant receptor proteins are involved in the signaling networks required for defense against pathogens. The novel pepper pathogen-induced gene CaMRP1 was isolated from pepper leaves infected with Xanthomonas campestris pv. vesicatoria (Xcv). This gene is predicted to encode a membrane-located receptor-like protein that has an N-terminal signal peptide and a C-terminal transmembrane helix. A CaMRP1-GFP fusion protein localized primarily to the plasma membrane of plant cells. Strong and early induction of CaMRP1 expression occurred following exposure of pepper plants to Xcv, Colletotricum coccodes, methyl jasmonate (MeJA) and wounding stress. Virus-induced gene silencing (VIGS) of CaMRP1 in pepper conferred enhanced basal resistance to Xcv infection, accompanied by induction of genes encoding basic PR1 (CaBPR1), defensin (CaDEF1) and SAR8.2 (CaSAR82A). In contrast, CaMRP1 overexpression (OX) in transgenic Arabidopsis plants resulted in increased disease susceptibility to Hyaloperonospora parasitica infection. Arabidopsis plants overexpressing CaMRP1 exhibited insensitivity to MeJA by causing reduced expression of MeJA-responsive genes. Overexpression also resulted in tolerance to NaCl and during salt stress, the expression of several abscisic acid-responsive genes was induced. Together, these results suggest that pepper CaMRP1 may belong to a new subfamily of membrane-located receptor-like proteins that regulate disease susceptibility, MeJA-insensitivity and salt tolerance. PMID- 18427934 TI - Spatial correlation between the prevalence of transmissible spongiform diseases and British soil geochemistry. AB - Transmissible spongiform encephalopathies (TSEs) are a group of fatal neurological conditions affecting a number of mammals, including sheep and goats (scrapie), cows (BSE), and humans (Creutzfeldt-Jakob disease). The diseases are widely believed to be caused by the misfolding of the normal prion protein to a pathological isoform, which is thought to act as an infectious agent. Outbreaks of the disease are commonly attributed to contaminated feed and genetic susceptibility. However, the implication of copper and manganese in the pathology of the disease, and its apparent geographical clustering, have prompted suggestions of a link with trace elements in the environment. Nevertheless, studies of soils at regional scales have failed to provide evidence of an environmental risk factor. This study uses geostatistical techniques to investigate the correlations between the distribution of TSE prevalence and soil geochemical variables across the UK according to different spatial scales. A similar spatial pattern in scrapie and BSE occurrence is identified, which may be linked with increasing pH and total organic carbon, and decreasing iodine concentration. However, the pattern also resembles that of the density of dairy farming. Nevertheless, despite the low spatial resolution of the TSE data available for this study, the fact that significant correlations are detected indicates there is a possibility of a link between soil geochemistry, scrapie, and BSE. It is suggested that further investigations of the prevalence of TSE and environmental exposure to trace metals should take into account the factors affecting their bioavailability. PMID- 18427935 TI - Biosorption of Cd(II) and Pb(II) onto brown seaweed, Lobophora variegata (Lamouroux): kinetic and equilibrium studies. AB - The present work deals with the biosorption performance of raw and chemically modified biomass of the brown seaweed Lobophora variegata for removal of Cd(II) and Pb(II) from aqueous solution. The biosorption capacity was significantly altered by pH of the solution delineating that the higher the pH, the higher the Cd(II) and Pb(II) removal. Kinetic and isotherm experiments were carried out at the optimal pH 5.0. The metal removal rates were conspicuously rapid wherein 90% of the total sorption occurred within 90 min. Biomass treated with CaCl(2) demonstrated the highest potential for the sorption of the metal ions with the maximum uptake capacities i.e. 1.71 and 1.79 mmol g(-1) for Cd(II) and Pb(II), respectively. Kinetic data were satisfactorily manifested by a pseudo-second order chemical sorption process. The process mechanism consisting of both surface adsorption and pore diffusion was found to be complex. The sorption data have been analyzed and fitted to sorption isotherm of the Freundlich, Langmuir, and Redlich-Peterson models. The regression coefficient for both Langmuir and Redlich Peterson isotherms were higher than those secured for Freundlich isotherm implying that the biosorption system is possibly monolayer coverage of the L. variegata surface by the cadmium and lead ions. FT-IR studies revealed that Cd(II) and Pb(II) binding to L. variegata occurred primarily through biomass carboxyl groups accompanied by momentous interactions of the biomass amino and amide groups. In this study, we have observed that L. variegata had maximum biosorption capacity for Cd(II) and Pb(II) reported so far for any marine algae. PMID- 18427936 TI - Human perception and awareness of ticks in a South Australian rural community and implications for management of Amblyomma triguttatum triguttatum. AB - The ornate kangaroo tick, Amblyomma triguttatum triguttatum, is invasive on Yorke Peninsula, South Australia. To assess the potential for developing comprehensive management strategies for the species, we mailed 1,000 surveys to Yorke Peninsula residents to investigate community perceptions of ticks, gather information on tick-host ecology, and assess whether or not occupation and length of residence affected whether residents had seen ticks. The response rate was 32%; most respondents (63%) had not seen ticks on Yorke Peninsula. Of those who had (n = 104), 56% had seen ticks on animals, especially lizards (31%) and kangaroos (20%); 50% had seen ticks on humans. Of the 64 respondents who believed places on Yorke Peninsula had many ticks, 56% identified Innes National Park and 75% areas on southern Yorke Peninsula. The likelihood of seeing ticks was not related to occupation. Whether respondents had seen ticks was related to length of residence on Yorke Peninsula (<10 years = 23.6%, 10-30 years = 44.0%, 31-50 years = 47.6%, 51-70 years = 30.9%, 71+ years = 21.4%). The number of respondents who had seen ticks on humans suggests that tick-borne diseases could become a public health issue on Yorke Peninsula. This finding is of particular concern because A. t. triguttatum is implicated in the epidemiology of Q fever in Queensland. Finally, respondents were unaware of the responsibility for tick management, which demonstrates a need for collaboration amongst stakeholders. PMID- 18427937 TI - The tick fauna of Sulawesi, Indonesia (Acari: Ixodoidea: Argasidae and Ixodidae). AB - Twenty-six species of ticks are reported from the island of Sulawesi (Celebes), Indonesia. These include two species of soft ticks (Argasidae), Carios batuensis and C. vespertilionis, and the following 24 species of hard ticks (Ixodidae): Amblyomma babirussae, A. breviscutatum, A. cordiferum, A. fimbriatum, A. helvolum, A. testudinarium, A. trimaculatum, A. varanense, Dermacentor atrosignatus, D. steini, Haemaphysalis celebensis, H. hystricis, H. kadarsani, H. papuana, H. psalistos, H. renschi, H. toxopei, H. wellingtoni, Ixodes cordifer, I. granulatus, Rhipicephalus haemaphysaloides, R. (Boophilus) microplus, R. pilans and R. sanguineus. This represents an almost three-fold increase in the number of tick species recorded (9) from Sulawesi since the last available list in 1950. The tick records reported herein represent a culmination of data from specimens in the U.S. National Tick Collection, new records of ticks from endemic tarsiers and associated vertebrates, and literature reviews. Collectively, the tick fauna of Sulawesi shows most affinities with the fauna of southeast Asia but there are distinct faunal elements that show relationships with other Indonesian islands, the Philippines or Australasia, as well as a few tick species with widespread geographical distributions. Some ticks known from Sulawesi have known or potential medical-veterinary significance. These include R. (B.) microplus which is a significant pest of cattle and a vector of the agents of bovine anaplasmosis, I. granulatus which is a vector of Langat virus and Lyme disease spirochetes and has been shown to harbor pathogenic rickettsiae in other parts of its range, and R. sanguineus which is a globally widespread ectoparasite of canines and a vector of canine pathogens and parasites. PMID- 18427939 TI - Comet assay: a reliable tool for the assessment of DNA damage in different models. AB - New chemicals are being added each year to the existing burden of toxic substances in the environment. This has led to increased pollution of ecosystems as well as deterioration of the air, water, and soil quality. Excessive agricultural and industrial activities adversely affect biodiversity, threatening the survival of species in a particular habitat as well as posing disease risks to humans. Some of the chemicals, e.g., pesticides and heavy metals, may be genotoxic to the sentinel species and/or to non-target species, causing deleterious effects in somatic or germ cells. Test systems which help in hazard prediction and risk assessment are important to assess the genotoxic potential of chemicals before their release into the environment or commercial use as well as DNA damage in flora and fauna affected by contaminated/polluted habitats. The Comet assay has been widely accepted as a simple, sensitive, and rapid tool for assessing DNA damage and repair in individual eukaryotic as well as some prokaryotic cells, and has increasingly found application in diverse fields ranging from genetic toxicology to human epidemiology. This review is an attempt to comprehensively encase the use of Comet assay in different models from bacteria to man, employing diverse cell types to assess the DNA-damaging potential of chemicals and/or environmental conditions. Sentinel species are the first to be affected by adverse changes in their environment. Determination of DNA damage using the Comet assay in these indicator organisms would thus provide information about the genotoxic potential of their habitat at an early stage. This would allow for intervention strategies to be implemented for prevention or reduction of deleterious health effects in the sentinel species as well as in humans. PMID- 18427938 TI - Folding and regulation in myosins II and V. AB - The enzymatic activity of many myosins is regulated by various means including calcium binding, phosphorylation or binding of receptor molecules. In this review we compare and contrast the regulation of smooth muscle myosin II and myosin Va with particular emphasis on the structural basis for the regulation. Both myosins adopt folded compact conformations in their off states, but the details of the conformations are markedly different. In the regulated smooth muscle myosin II, the key feature is an asymmetric interaction between the two heads of the molecule with contributions of specific tail-head interactions. In myosin V the key feature is an interaction between the heads and the globular tail domain. PMID- 18427940 TI - Exploring emerging issues in research higher degree supervision of professional doctorate students in the health sciences. AB - Over the past 20 years, there has been an increase in the number of students undertaking traditional research masters and doctor of philosophy courses in the health sciences. This paper reviews and summarizes several aspects of the literature related to supervision of students in these traditional research higher degree (RHD) programs in Australasia: elements of RHD supervision; use of information and communication technologies in RHD supervision; warning signs for non-completion of RHD candidature; and evaluation of RHD supervision. The growth in traditional RHD enrolments has been paralleled by similar expansion in professional doctorate programs, and this paper draws attention to issues relating to supervision of these professional doctorate students, especially those studying at a distance. It draws on earlier research which compared structures of such programs across Australia and grounded professional doctorate supervision firmly in the workplace. Further, the emerging small body of literature associated with professional doctorate supervision is used to highlight features shared with and those distinct from traditional RHD supervision, and to identify fertile topics for further research into this burgeoning area. PMID- 18427941 TI - Reflections on experimental research in medical education. AB - As medical education research advances, it is important that education researchers employ rigorous methods for conducting and reporting their investigations. In this article we discuss several important yet oft neglected issues in designing experimental research in education. First, randomization controls for only a subset of possible confounders. Second, the posttest-only design is inherently stronger than the pretest-posttest design, provided the study is randomized and the sample is sufficiently large. Third, demonstrating the superiority of an educational intervention in comparison to no intervention does little to advance the art and science of education. Fourth, comparisons involving multifactorial interventions are hopelessly confounded, have limited application to new settings, and do little to advance our understanding of education. Fifth, single-group pretest-posttest studies are susceptible to numerous validity threats. Finally, educational interventions (including the comparison group) must be described in detail sufficient to allow replication. PMID- 18427942 TI - Exploring continuous clinical placement for undergraduate students. AB - Clinical placements are integral to health professional preparatory courses. These placements allow for the application of classroom-based learning into real patient care situations. In doing so, they provide opportunities for applying theoretical knowledge into practice contexts, skills development and socialisation into the chosen profession. However, despite its recognised importance across health professions, little has been written about optimal structures for supporting clinical learning. This paper presents one group of findings from a larger qualitative study aimed at exploring health professional student's clinical experiences and their impact on career intentions. Findings reported here present a group of undergraduate midwifery student's perspectives on a "home" hospital clinical placement model where continuous clinical placements were taken in the same agency (or hospital group) for 2 days per week over the final 2 years of their course. Two main themes emerged from the data analysis, these being, 'familiarity' and 'continuity'. It is concluded that continuous placements in the same clinical setting have the potential to offer greater opportunities for learning and early professional socialisation than traditional block (Monday to Friday) placements. They can offer a more integrated approach to classroom theory and its application into practice. PMID- 18427943 TI - An insight into vaginal surgery in a severely masculinized CAH patient. AB - Reconstruction and exteriorization of the "high" vagina in girls with congenital adrenal hyperplasia (CAH) is technically challenging. Although current surgical thinking favors correction in infancy the evidence from several long-term studies indicates that many patients will require further surgery in adolescence or adult life. Deferred vaginoplasty has been advocated because the procedure may be technically easier as a consequence of changes in the characteristics of the genital tissues and vaginal anatomy. The role of urogenital mobilization is as yet undetermined but carries promise for the short, high vagina. We describe a patient where the radiological and clinical findings support the argument for deferred vaginoplasty. PMID- 18427944 TI - Comparison of sleep quality between hemodialysis and peritoneal dialysis patients. AB - AIM: Sleep disorders are common in patients with end-stage renal disease. Although studies have been conducted on the type and frequency of sleep disturbances in hemodialysis and peritoneal dialysis patients, there has been no study comparing the sleep quality between these two groups. Therefore, we aimed to compare sleep quality between hemodialysis and peritoneal dialysis patients. METHODS: A total of 102 patients (52 hemodialysis and 50 peritoneal dialysis) were included in the study. The Pittsburgh sleep quality index (PSQI) was used for the assessment of sleep quality. Two groups were compared for seven components of the PSQI questionnaire and global score as well as for clinical and laboratory findings. We also assessed the independent predictors of sleep quality. RESULTS: There were 51 male and 51 female patients (29 male and 23 female in hemodialysis group versus 22 male and 28 female in peritoneal dialysis group). The mean age was 55.5+/-14.6 years in the hemodialysis and 51.5+/-18.1 years in the peritoneal dialysis group. The median dialysis duration was 36 (77.0) months. The sleep quality was poor in 88.5% of the hemodialysis patients and 78.0% of the peritoneal dialysis patients. However, this difference in sleep quality was not significant between the two groups (P>0.05). There was a significant association between the sleep quality and the age, presence of diabetes mellitus, and serum albumin. Among these variables, only age was found to be an independent predictor of sleep quality. CONCLUSIONS: Hemodialysis and peritoneal dialysis patients had a similar high rate of poor sleep quality. Further studies are necessary to investigate the causes of poor quality of sleep and to investigate methods to improve sleep quality in this population. PMID- 18427945 TI - Perceived age as a biomarker of ageing: a clinical methodology. AB - In a previous field-based study, how old one looks for one's age (perceived age) was found to be predictive of mortality in elderly individuals. In conjunction, perceived age is of relevance and interest to the layperson. Here, a clinical methodology for generating perceived age as a biomarker of facial ageing is detailed. The methodology utilises facial photographs of subjects to present images to large numbers of age assessors who are primarily nationals of the country of study origin. In five observational studies in five different countries involving 874 female subjects it was found that subject age and assessor gender, nationality, age and ageing expertise had little effect on the perceived age data generated. However, increasing the numbers of age assessors up to 50 substantially increased the reproducibility of the mean perceived age for an image and a minimum of 10 assessors were required to give reproducible data. This methodology was also compared to a methodology that utilises passport-type photographs of subjects typically taken in field studies. Although the perceived age data from the two types of images were more similar to each other than to chronological age, there was a marked difference between the two sets of data. Therefore, to allow meaningful comparisons across perceived age studies, the same type of image should be used for the generation of perceived age. In conclusion, the methodology detailed here has demonstrated that perceived age can be a reproducible measure when large numbers of adult age assessors are used and can be utilised globally in studies to investigate facial ageing. PMID- 18427946 TI - The pairing center plays a key role in homolog paring: an explanation for adjacent-2 segregation in interchange heterozygotes. AB - Having reflected on the discrepancy between various views of chromosome behavior during meiosis, we propose an alternative description of Mendel's first law of segregation by referring to the segregation of pairing centers instead of centromeres. We also propose an alternative description of Mendel's second law of independent assortment, which refers to the free combination of different pairing centers. This interpretation is based on the modified concept that true 'homologous chromosomes' should carry the pairing center rather than centromere: the length of homology or the importance of the homologous segment on the chromosome is the crucial factor in homologous chromosome pairing and synapsis. PMID- 18427947 TI - The correlations between HPV16 infection and expressions of c-erbB-2 and bcl-2 in breast carcinoma. AB - Breast carcinoma (BC) is a prevalent malignant tumour occurring in women. Many studies have indicated the role of human papilloma virus type 16 (HPV16) in the pathogenesis of BC; however, the correlations of HPV16 infection with the clinicopathologic features of BC and the expressions of c-erbB-2 and bcl-2 have not yet been elucidated. In this study, HPV16 was detected by amplifying the HPV16 E6 gene by the polymerase chain reaction method, and the expressions of c erbB-2 and bcl-2 in 40 BCs and 20 normal breast tissue samples, obtained from Shaanxi Province, were examined using the streptavidin-peroxidase method with monoclonal antibodies specific to c-erbB-2 and bcl-2. The infection rate of HPV16 E6 and the positive expression rate of c-erbB-2 were significantly higher in the BCs than in the normal tissues (HPV16 E6: 60% vs. 5%; c-erbB-2: 42.5% vs. 5%, P < 0.05). However, the positive expression rate of bcl-2 was significantly lower in the BCs than in the normal tissues (67.5% vs. 95%, P < 0.05). The infection rate of HPV16 did not correlate with any of the pathological features observed (P > 0.05). HPV16 infection correlated with bcl-2 expression (P = 0.015) but not with c-erbB-2 expression (P = 0.747) in the BCs. Interestingly, HPV16 infection correlated with bcl-2 expression in grade I BCs (P = 0.018) but not in grade II III BCs (P = 0.633). Our data suggest that HPV16 infection is correlated with bcl 2 expression in BCs. PMID- 18427948 TI - Quality of life in cancer patients in South Asia: psychometric properties of the Sinhala version of the EORTC QLQ-C30 in cancer patients with heterogeneous diagnoses. AB - BACKGROUND: The European Organisation for Research and Treatment of Cancer Quality of Life Core Questionnaire (EORTC QLQ-C30) was evaluated for its psychometric properties in a sample of cancer patients from the culturally distinct South Asian subcontinent, which accounts for a significant proportion of the global cancer burden. METHODS: Psychometric testing assessed the hypothesised scale structure, internal consistency, content and construct validity, and acceptability of the Sinhala version of the QLQ-C30 independently in two heterogeneous groups of cancer patients at pretreatment (N = 489) and during treatment (N = 343). RESULTS: Qualitative feedback from an expert panel assessing content validity recommended measuring family support during illness as an additional, culturally-relevant dimension of health-related quality of life (HRQL). Compliance was high (100%), with little missing data (0.11%). Multitrait scaling results supported the scale structure of the QLQ-C30, with the exception of the cognitive functioning scale, which was also the only scale that did not meet the 0.70 internal consistency criteria in either sample. Interscale correlations were of a moderate size, with conceptually related scales showing higher correlations. All scales were able to discriminate clearly between pre- and current treatment patients (P < 0.01), although results were less consistent when comparing groups formed on the basis of age and disease stage. CONCLUSIONS: Overall psychometric results confirmed the QLQ-C30 as a reliable and valid questionnaire for assessing HRQL of cancer patients in Sri Lanka. PMID- 18427949 TI - The impact of chronic hepatitis C and co-morbid illnesses on health-related quality of life. AB - OBJECTIVES: Determine the relative impact of chronic hepatitis C (CHC) and co morbid illnesses on health-related quality of life (HRQoL) in 3023 randomly selected veterans with known hepatitis C virus antibody (anti-HCV) status who previously completed a veteran-specific HRQoL questionnaire (SF-36V). METHODS: Multiple regression analyses were performed to measure the relative contribution of anti-HCV status, four demographic variables, and ten common medical and six psychiatric co-morbidities to HRQoL between 303 anti-HCV(+) and 2720 anti-HCV(-) patients. RESULTS: Anti-HCV(+) veterans were younger, reported a lower HRQoL on seven of eight 36-Item Short Form Health Survey for Veterans (SF-36V) subscales (P < or = 0.001) and the mental component summary (MCS) scale (P < 0.001). The ten medical and six psychiatric co-morbidities had variable impact on predicting lower HRQoL in both groups. After adjusting for demographic variables and co morbid illnesses, we found that anti-HCV(+) patients reported a significantly lower MCS score (P < 0.001) and a trend toward a lower physical component summary (PCS) score (P < 0.07) compared to anti-HCV(-) veterans. Among the anti-HCV(+) veterans, co-morbid medical illnesses contributed to impaired PCS but not to MCS. CONCLUSIONS: Veterans with CHC were younger than HCV(-) veterans and hence less likely to have other co-morbid medical illnesses. Medical co-morbidities seen in those veterans with CHC contribute to impaired PCS but not MCS. Anti-HCV(+) status negatively affects HRQoL, particularly MCS, independently of medical and psychiatric co-morbidities. PMID- 18427950 TI - Measuring health-related quality of life for persons with mobility impairments: an enabled version of the short-form 36 (SF-36E). AB - OBJECTIVE: Examine psychometric properties of the SF-36 and SF-36E for mobility impaired individuals and assess whether the SF-36E yields higher health-related quality-of-life scores. METHODS: We altered the SF-36 Physical Function scale by substituting the word "go" for "walk" and "climb" and changed the stem to reflect function using assistive devices. We compared responses between versions for 201 individuals with disabilities (n = 95 wheelchair users, n = 48 other device users, 58 = no device users). RESULTS: Both surveys yielded reliable scores, but floor and ceiling effects occurred with both versions. Confirmatory factor analyses demonstrate good fit for the SF-36 and SF36E, but were compromised by low sample size. Respondents demonstrated significantly better Role Physical, Bodily Pain, and Vitality on the SF-36E, but worse General Health. CONCLUSIONS: The World Health Organization framework, the International Classification of Functioning, Disability, and Health (ICF) suggests that it is important to know both what one can do without assistance (capacity) and what one can do with assistance (performance). Results suggest that the SF-36E successfully measures performance among mobility-impaired individuals, including wheelchair users. However, further validation studies of the SF-36 and SF-36E are warranted with samples of individuals with disabilities. PMID- 18427951 TI - The use of focus groups in the development of the PROMIS pediatrics item bank. AB - PURPOSE: To understand differences in perceptions of patient-reported outcome domains between children with asthma and children from the general population. We used this information in the development of patient-reported outcome items for the Patient-Reported Outcomes Measurement Information System Pediatrics project. METHODS: We conducted focus groups composed of ethnically, racially, and geographically diverse youth (8-12, 13-17 years) from the general population and youth with asthma. We performed content analysis to identify important themes. RESULTS: We identified five unique and different challenges that may confront youth with asthma as compared to general population youth: (1) They experience more difficulties when participating in physical activities; (2) They may experience anxiety about having an asthma attack at anytime and anywhere; (3) They may experience sleep disturbances and fatigue secondary to their asthma symptoms; (4) Their health condition has a greater effect on their emotional well being and interpersonal relationships; and (5) Youth with asthma report that asthma often leaves them with insufficient energy to complete their school activities, especially physical activities. CONCLUSIONS: The results confirm unique experiences for children with asthma across a broad range of health domains and enhance the breadth of all domains when creating an item bank. PMID- 18427952 TI - Comparison of the cardiac electrophysiological effects between doxazosin and bunazosin. AB - In Langendorff-perfused adult rat heart with constant pressure at 80 mmHg, we found doxazosin, an alpha(1) adrenoceptor blocker, at 10 muM prolonged PR interval and induced occasional arrhythmia followed by complete inhibition of the sinus rhythm, whereas bunazosin, another alpha(1)-blocker, at same concentration did not. The results of voltage-clamp study showed that, at the concentration of 10 muM, doxazosin inhibited I (Na), I ( (Ca,L) ), I (to), and Iss without changing I (k1) but bunazosin only inhibited I ( (Ca,L) ) about 30%. Doxazosin also caused markedly negative shift of the I (Na) inactivation curve. In current clamp study, doxazosin prolonged action potential duration in association with the decreased action potential amplitude and upstroke velocity, whereas bunazosin did not. We hypothesize that doxazosin-induced arrhythmia may result from the heterogeneous or different level of I ( (Ca,L) ) blockade of AV nodal tissue. In conclusion, the present study suggests that bunazosin is safer than doxazosin for long-term treatment in view of electrophysiological effect. However, the underlying mechanism of doxazosin induced nodal arrhythmia is still needed to be determined. PMID- 18427953 TI - Philosophy of technology and macro-ethics in engineering. AB - The purpose of this paper is to diagnose and analyze the gap between philosophy of technology and engineering ethics and to suggest bridging them in a constructive way. In the first section, I will analyze why philosophy of technology and engineering ethics have taken separate paths so far. The following section will deal with the so-called macro-approach in engineering ethics. While appreciating the initiative, I will argue that there are still certain aspects in this approach that can be improved. In the third, fourth, and fifth sections, I will point out three shortcomings of engineering ethics in terms of its macro level discourse and argue that a number of certain insights taken from the study of philosophy of technology could be employed in overcoming those problems. In the concluding section, a final recommendation is made that topics of philosophy of technology be included in the curriculum of engineering ethics. PMID- 18427954 TI - Engineering ethics in Puerto Rico: issues and narratives. AB - This essay discusses engineering ethics in Puerto Rico by examining the impact of the Colegio de Ingenieros y Agrimensores de Puerto Rico (CIAPR) and by outlining the constellation of problems and issues identified in workshops and retreats held with Puerto Rican engineers. Three cases developed and discussed in these workshops will help outline movements in engineering ethics beyond the compliance perspective of the CIAPR. These include the Town Z case, Copper Mining in Puerto Rico, and a hypothetical case researched by UPRM students on laptop disposal. The last section outlines four future challenges in engineering ethics pertinent to the Puerto Rican situation. PMID- 18427955 TI - Liberty to decide on dual use biomedical research: an acknowledged necessity. AB - Humanity entered the twenty-first century with revolutionary achievements in biomedical research. At the same time multiple "dual-use" results have been published. The battle against infectious diseases is meeting new challenges, with newly emerging and re-emerging infections. Both natural disaster epidemics, such as SARS, avian influenza, haemorrhagic fevers, XDR and MDR tuberculosis and many others, and the possibility of intentional mis-use, such as letters containing anthrax spores in USA, 2001, have raised awareness of the real threats. Many great men, including Goethe, Spinoza, J.B. Shaw, Fr. Engels, J.F. Kennedy and others, have recognized that liberty is also a responsibility. That is why the liberty to decide now represents an acknowledged necessity: biomedical research should be supported, conducted and published with appropriate measures to prevent potential "dual use". Biomedical scientists should work according to the ethical principles of their Code of Conduct, an analogue of Hippocrates Oath of doctors; and they should inform government, society and their juniors about the problem. National science consulting boards of experts should be created to prepare guidelines and control the problem at state level. An international board should develop minimum standards to be applicable by each country. Bio-preparedness is considered another key-measure. PMID- 18427956 TI - A revision of the family Dissonidae Kurtz, 1924 (Copepoda: Siphonostomatoida). AB - Two new species of the parasitic copepod genus Dissonus Wilson, 1906 are described: D. excavatus n. sp. from the gills of a labrid, Bodianus perditio, and a lutjanid, Macolor niger, collected off New Caledonia and Taiwan, and D. inaequalis n. sp. from a hemiscylliid elasmobranch, Chiloscyllium punctatum, collected off Sarawak (Malaysia) and the Philippines. Material of D. heronensis Kabata, 1966 is described from a balistid host, Pseudobalistes fuscus, off New Caledonia, and this constitutes a new host record for this parasite. D. manteri Kabata, 1966 was collected from four serranid host species off New Caledonia and from one of the same hosts off Taiwan. Two of the hosts from New Caledonia, Plectropomus laevis and Epinephelus cyanopodus, represent new host records. D. pastinum Deets & Dojiri, 1990 was recognised as a new synonym of D. nudiventris Kabata, 1966, so the total number of valid species is now twelve. Material from museum collections of D. nudiventris, D. similis Kabata, 1966 and D. spinifer Wilson, 1906 was re-examined and provided new information which is utilised in a key to all valid species of Dissonus. PMID- 18427957 TI - Cystacanths of Acanthocephala in notothenioid fish from the Beagle Channel (sub Antarctica). AB - The morphology of relaxed cystacanths of polymorphid acanthocephalans collected from notothenioid fishes in the Beagle Channel (Magellanic subregion of sub Antarctica) is described. A parasite of birds, Andracantha baylisi (Zdzitowiecki, 1986), was found in Patagonotothen longipes and Champsocephalus esox. It has: a proboscis 0.82-0.89 mm long; a proboscis hook formula of 16 rows of 9/10-10/11, including 4-5 basal hooks; distal hooks with the longest blades; a fore-trunk not separated from the hind-trunk by a constriction; large somatic spines arranged in two zones separated by a zone of small, loosely dispersed spines; and only the anterior 36-40% of ventral side of the trunk is covered with spines. One male specimen of Corynosoma sp. was found in Patagonotothen tessellata. It differs from A. baylisi in that the distal proboscis hooks are similar in length to the prebasal hooks, it has a smaller proboscis (0.77 mm) and in the distribution of the somatic spines, which are contiguous with the genital spines on the ventral side of the trunk and lack a zone of small spines between zones of larger spines. A parasite of seals and fur seals, Corynosoma evae Zdzitowiecki, 1984, was found in P. longipes and Champsocephalus esox. It has: a proboscis 0.61-0.78 mm long; a proboscis hook formula of 20-22 rows of 12-13, including 3/4-4 basal hooks; prebasal hooks with the longest blades; a trunk divided into fore-trunk and hind trunk; somatic spines covering the anterior 64-74% of the ventral side of the trunk; genital spines present only in males; and a terminal genital opening in both sexes. Corynosoma beaglense n. sp. was found in Champsocephalus esox. It has: an almost cylindrical proboscis (length 0.52-0.56 mm); a proboscis hook formula of 16 rows of 9/10-10/11, including 4-4/5 basal hooks; distal hooks shorter than the prebasal hooks; a fore-trunk not separated from the hind-trunk by a constriction; somatic spines contiguous with the genital spines on the ventral side of the trunk of the male and covering the entire length of the ventral side of the female trunk, and the presence of genital spines surrounding the terminal genital pore of the male. The definitive host of this species is unknown. PMID- 18427958 TI - Redescription and new data on Microsomacanthus diorchis (Fuhrmann, 1913) (Cestoda: Hymenolepididae). AB - Microsomacanthus diorchis (Fuhrmann, 1913) is redescribed and illustrated on the basis of the type-material and new findings from common eider Somateria mollissima captured in Iceland and specimens from the same host species from the Barents, White and Bering Seas. A lectotype is designated and an amended diagnosis is provided. The main differentiating features of M. diorchis are the size and shape of rostellar hooks and the cirrus, the well-marked delay in the antiporal testis development and the bow-shaped uterus. This parasite is shown to be specific to S. mollissima for both Atlantic and Pacific populations of the host. Hymenolepis (Microsomacanthus) somateriae of Bishop & Threlfall (1974) [nec M. somateriae Ryzhikov, 1965] is recognised as a synonym of M. diorchis (Fuhrmann, 1913). The taxonomic position of the species described as Aploparaksis murmanica Baylis, 1919 from common eider is discussed. PMID- 18427959 TI - A new species of Potamotrygonocestus Brooks & Thorson, 1976 (Eucestoda: Tetraphyllidea) from Plesiotrygon iwamae Rosa, Castello & Thorson (Mylliobatoidea: Potamotrygonidae) and a redescription of Potamotrygonocestus chaoi Marques, Brooks & Araujo, 2003. AB - Potamotrygonocestus Brooks & Thorson, 1976 is currently represented by six recognised species of tetraphyllidean cestodes inhabiting Neotropical freshwater stingrays. Potamotrygonid stingrays examined to date have included only a single specimen of Plesiotrygon iwamae. Only one species of tetraphyllidean, Potamotrygonocestus chaoi Marques, Brooks & Araujo, 2003, has been described from this host, and this description was based on limited material. New efforts to document the diversity in this host species resulted in the collection of eight additional specimens of P. iwamae, one of these from the upper Rio Solimoes, at Sao Paulo de Olivenca Amazonas during September, 2003, and seven from the Baia do Marajo, Para, during November, 2003. The specimen from the upper Solimoes was found to be infected with P. chaoi. Voucher material from this stingray was used for the redescription of this cestode, which is characterized by strobila 8.78 22.83 mm long and a great number of proglottides, 58-93; the new material provided strobilar length and proglottis counts for complete worms. Potamotrygonocestus marajoara n. sp. is the second species of this genus reported from Plesiotrygon iwamae, although it appears to be restricted to the lower Amazon. This new species resembles P. chaoi in possessing filitriches and blade like spinitriches on the scolex, cephalic peduncle and cirrus, but differs from other species of the genus in the number of testes, which is 44 on average per proglottis, and by having apical sucker measuring 95-175 microm in length. Additional data on the distribution and morphology of the microtriches and a detailed description of the female reproductive system are also provided in this study. PMID- 18427960 TI - Neoskrjabinolepis nuda n. sp. from shrews on Sakhalin Island, Russia, with a taxonomic review of Neoskrjabinolepis Spasskii, 1947 (Cestoda: Cyclophyllidea: Hymenolepididae). AB - Neoskrjabinolepis (Neoskrjabinolepidoides) nuda n. sp. is described from the shrews Sorex unguiculatus (type-host), S. gracillimus, S. isodon and S. caecutiens on Sakhalin Island, Russia. The new species is characterised by: rostellar hooks 40-44 microm long and provided with small epiphyseal thickening of the handle; a long (95-100 microm) cirrus consisting of basal region with claw shaped spines, a parabasal region with thin needle-shaped spines and an unarmed distal region; a cirrus-sac extending well into the median field; and 15-22 eggs per gravid uterus. A review of the species of Neoskrjabinolepis Spasskii, 1947 is presented. Currently, this genus includes nine species and is divided in two subgenera on the basis of strobilar development, which is gradual in the subgenus Neoskrjabinolepis (four species) and serial in the subgenus Neoskrjabinolepidoides Kornienko, Gulyaev & Mel'nikova, 2006 (five species). An amended generic diagnosis and an identification key to Neoskrjabinolepis spp. are presented. PMID- 18427961 TI - Chemoprevention of pancreatic cancer: characterization of Par-4 and its modulation by 3,3' diindolylmethane (DIM). AB - PURPOSE: Cancer chemoprevention is defined as the use of natural, synthetic, or biological agents to suppress, reverse or prevent the carcinogenic process from turning into aggressive cancer. Prostate apoptosis response-4 (Par-4) is a unique pro-apoptotic protein that selectively induces apoptosis in prostate cancer cells. However, its role in other malignancies has not been fully explored. This study tries to identify the functional significance of Par-4 in pancreatic cancer. METHODS: Multiple molecular techniques such as Western blot analysis, trypan blue assay for cell viability, MTT assay for cell growth inhibition and Histone/DNA ELISA for apoptosis were used. RESULTS: Western blot analysis revealed that 3,3'-diindolylmethane (DIM) a chemopreventive agent, specifically its more bioavailable formulation, B-DIM, at low doses (20 micromol/L) induces Par-4, in L3.6pl and Colo-357 pancreatic cancer cells. At similar doses, DIM reduced cell viability and caused cell growth inhibition and apoptosis. Moreover, DIM pre-treatment sensitized the cells to cytotoxic action of chemotherapeutic drug gemcitabine through up-regulation of Par-4. CONCLUSION: The induction of Par 4 is indirectly related to increased sensitivity and cell death through apoptosis. To our knowledge the results reported here showed, for the first time, the induction of Par-4 by chemopreventive agents, in general, and DIM, in particular, in pancreatic cancer cells in vitro. PMID- 18427962 TI - Different curcuminoids inhibit T-lymphocyte proliferation independently of their radical scavenging activities. AB - PURPOSE: We investigated the inhibitory effects of curcumin, curcumin derivatives and degradation products on OKT3-induced human peripheral blood mononuclear cell (PBMC) proliferation and the role of their radical scavenging activity. METHODS: OKT3-induced human PBMC proliferation was determined by measuring 3H-thymidine incorporation. Radical scavenging activity was evaluated by using an in vitro DPPH assay. RESULTS: OKT3-induced PBMC proliferation was inhibited by curcumin, isocurcumin, bisdesmethoxy-, diacetyl-, tetrahydro-, hexahydro-, and octahydrocurcumin as well as by vanillin, ferulic acid, and dihydroferulic acid with IC50-values of 2.8, 2.8, 6.4, 1.0, 25, 38, 82, 729, 457, and >1,000 microM, respectively. The investigated substances with the strongest effect on radical scavenging were tetrahydro-, hexahydro-, and octahydrocurcumin with IC50 values of 10.0, 11.7, and 12.3 microM, respectively. IC50-values of dihydroferulic acid, ferulic acid, and curcumin were 19.5, 37, and 40 microM. The substances with the lowest radical scavenging activities were vanillin, isocurcumin, diacetylcurcumin, and bisdesmethoxycurcumin with IC50 values higher than 100 microM each. CONCLUSIONS: Curcuminoid-induced inhibition of OKT3-induced PBMC proliferation depends on the number of carbon atoms and double bonds of the 1,6 heptadiene-3,5-dione structure as well as on the phenolic ring substitutes of the curcuminoids but is not correlated to their respective radical scavenging activity. PMID- 18427963 TI - Hepatoprotective effect of infliximab, an anti-TNF-alpha agent, on carbon tetrachloride-induced hepatic fibrosis. AB - To assess the effect of infliximab, an anti-tumor necrosis factor (TNF)-alpha agent, on the carbon tetrachloride (CCl(4))-induced hepatic fibrosis in rats. Rats were randomized into three groups (n=9). The control group received only intraperitoneal (i.p.) olive oil. Hepatic fibrosis was induced by repeated i.p. injections of 1.5 ml/kg CCl(4) (1:3 mixture with olive oil) for 5 weeks in the remaining two groups which were also injected subcutaneous saline or 2 mg/kg infliximab. Infliximab reduced the levels of aspartate aminotransferase and alanine aminotransferase (p<0.05 for both). The scores of hepatic necrosis, inflammation and fibrosis, and expression of alpha-smooth muscle actin were lower in the infliximab-treated group than the CCI(4)-treated group (p<0.01, p<0.001, p<0.01, p<0.001, respectively). However, there was no significant difference in terms of liver tissue and plasma malondialdehyde, and serum TNF-alpha levels, while infliximab relatively reduced the level of transforming growth factor beta(1) (373.0+/-153.1 vs. 280.8+/-127.1 pg/ml). Treatment with infliximab attenuated the necro-inflammation and fibrogenesis in the CCI(4)-induced hepatic fibrosis, and thus it might be effective as a therapeutic anti-fibrotic agent. PMID- 18427965 TI - Introduction to the Journal of Cross-Cultural Gerontology, special issue on aging and social change in Africa. PMID- 18427964 TI - Anti-inflammatory effects of ethanol extract from Kummerowia striata (Thunb.) Schindl on lps-stimulated RAW 264.7 cell. AB - Kummerowia striata (Thunb.) Schindl has long been used as a fork herb in inflammation-related therapy. This study was undertaken to determine the anti inflammatory effect of the plant. High performance liquid chromatography (HPLC) was used for evaluating the extract. While dexamethasone (DM) was used as a positive control, the effects of ethanol extract on the production of IL-1beta, IL-6, NO, COX-2 and TNF-alpha, the expression of iNOS mRNA, TNF-alpha mRNA, COX-2 mRNA, protein production of COX-2 and HO-1, NF-kappaB and I-kappaB of LPS stimulated RAW 264.7 cells were studied by sandwich ELISA, real-time PCR, Western blot analysis and immunocytochemistry assay respectively. The results showed that K. striata (Thunb.) Schindl had a good anti-inflammatory effect on LPS-stimulated RAW264.7 cell. On one hand, it could significantly inhibit the production of IL 1beta, IL-6, NO, TNF-alpha, COX-2 in LPS-stimulated cell than that of single LPS stimulated cell (p < 0.01 or p < 0.05). On the other hand, it could increase the production of IL-10 and HO-1 than that of single LPS intervention cell (p < 0.01 or p < 0.05). Furthermore, the extract also could inhibit the production of NF kappaB and I-kappaB compared to single LPS stimulated cell. In a word, it suggested that the anti-inflammatory actions of K. striata (Thunb.) Schindl ethanol extract might be due to the down-regulation of IL-1beta, IL-6, NO, TNF alpha and COX-2 via the suppression of NF-kappaB activation and conversation of I kappaB production, and another pathway was up regulating the production of IL-10 and HO-1. PMID- 18427966 TI - Fluctuation-driven rhythmogenesis in an excitatory neuronal network with slow adaptation. AB - We study an excitatory all-to-all coupled network of N spiking neurons with synaptically filtered background noise and slow activity-dependent hyperpolarization currents. Such a system exhibits noise-induced burst oscillations over a range of values of the noise strength (variance) and level of cell excitability. Since both of these quantities depend on the rate of background synaptic inputs, we show how noise can provide a mechanism for increasing the robustness of rhythmic bursting and the range of burst frequencies. By exploiting a separation of time scales we also show how the system dynamics can be reduced to low-dimensional mean field equations in the limit N --> infinity. Analysis of the bifurcation structure of the mean field equations provides insights into the dynamical mechanisms for initiating and terminating the bursts. PMID- 18427967 TI - Modeling of quantal neurotransmitter release kinetics in the presence of fixed and mobile calcium buffers. AB - The local calcium concentration in the active zone of secretion determines the number and kinetics of neurotransmitter quanta released after the arrival of a nerve action potential in chemical synapses. The small size of mammalian neuromuscular junctions does not allow direct measurement of the correlation between calcium influx, the state of endogenous calcium buffers determining the local concentration of calcium and the time course of quanta exocytosis. In this work, we used computer modeling of quanta release kinetics with various levels of calcium influx and in the presence of endogenous calcium buffers with varying mobilities. The results of this modeling revealed the desynchronization of quanta release under low calcium influx in the presence of an endogenous fixed calcium buffer, with a diffusion coefficient much smaller than that of free Ca(2+), and synchronization occurred upon adding a mobile buffer. This corresponds to changes in secretion time course parameters found experimentally (Samigullin et al., Physiol Res 54:129-132, 2005; Bukharaeva et al., J Neurochem 100:939-949, 2007). PMID- 18427968 TI - Identification of functional domains of phosphoproteins of two morbilliviruses using chimeric proteins. AB - The paramyxovirus P protein is an essential component of the transcriptase and replicase complex along with L protein. In this article, we have examined the functional roles of different domains of P proteins of two closely related morbilliviruses, Rinderpest virus (RPV) and Peste des petits ruminants virus (PPRV). The PPRV P protein physically interacts with RPV L as well as RPV N protein when expressed in transfected cells, as shown by co-immunoprecipitation. The heterologous L-P complex is biologically active when tested in a RPV minigenome replication/transcription system, only when used with PPRV N protein but not with RPV N protein. Employing chimeric PPRV/RPV cDNAs having different coding regions of P protein in the minigenome replication/transcription system, we identified a region between 290 and 346 aa in RPV P protein necessary for transcription of the minigenome. PMID- 18427969 TI - Characterization and genetic diversity of sugarcane streak mosaic virus causing mosaic in sugarcane. AB - Sixty-three sugarcane leaf samples were collected from fifty-eight sugarcane varieties, evolved from eleven major sugarcane growing states in India, Australia, South Africa and USA. In RT-PCR, using gene specific primers for sugarcane streak mosaic virus (SCSMV)-CP, 58 of 63 sugarcane samples were found positive to the virus infection and rest of the five samples were negative. Partial CP gene sequences of 42 SCSMV isolates including an isolate from aphid colony (Melanaphis indosacchari) infested on sugarcane variety from this study were characterized after cloning and sequencing for selective isolates represented by at least one isolate from each location. The new sequences identified in the study were named as SCSMV-CB isolates. Fifty two sequences including the 10 database sequences (complete CP cds) deposited earlier from this institute were compared with each other as well as GenBank database sequences of Potyviridae members viz., Rymovirus, Potyvirus, Ipomovirus, Tritimovirus and eight sequences of SCSMV reported from elsewhere. Among the SCSMV-CB isolates sequenced in the study, 85.7-100% (nucleotide) and 89.9-100% (amino acid) sequence identities were observed and with the other data base sequences of SCSMV, the respective identities were 82.2-97.5 and 89.7-98.6%. Grouping of the isolates by the maximum likelihood with molecular clock model, distributed 60 SCSMV sequences including the eight database sequences deposited by other SCSMV working groups from India and USA in 16 different phylogenetic groups. Although the isolates of SCSMV were relatively close to Ipomovirus and Tritimovirus, they were sandwiched between Rymovirus and Ipomovirus. The sequence comparison and phylogenetic studies revealed that the relatedness of SCSMV with the potyviral related genera was comparatively low to consider it as a member of earlier described potyviral genera, hence the genus "Susmovirus" (sugarcane streak mosaic virus) has been proposed, with SCSMV as the sole species to be included. The 52 SCSMV-CB isolates from this institute were distributed in 14 phylogenetic groups and the grouping pattern revealed that the virus isolates could not be grouped based on geographical origin of the host varieties or longevity of the host variety. PMID- 18427970 TI - Drug and HIV-related risk behaviors after geographic migration among a cohort of injection drug users. AB - To determine whether migration impacted on drug use and HIV-related risk behaviors among injection drug users (IDU), we identified participants in a prospective cohort of IDU (Vancouver Injection Drug User Study) who had reported migrating out of Greater Vancouver between May 1996 and November 2005. We compared risk behaviors before and after a move for individuals who migrated (movers) and for a similar period for non-movers using linear growth curve analyses. In total, 1,122 individuals were included, including 430 (38.3%) women and 331 (29.5%) Aboriginal participants. Among these, 192 (17.1%) individuals reported migrating out of Greater Vancouver between 1996 and 2005 while 930 (82.9%) did not. Movers were significantly younger than non-movers: 32.0 (Interquartile Range [IQR]: 24.3-39.2) and 34.6 (IQR: 26.9-40.8) respectively. A significant decrease in those reporting unstable housing, frequent heroin and cocaine injection occurred only in movers. Our findings suggest that, in this setting, risk-taking among IDU declines following periods of migration out of Greater Vancouver. PMID- 18427971 TI - Adolescent sexual risk: factors predicting condom use across the stages of change. AB - This study examined factors associated with high-risk adolescents' movement toward or away from adopting consistent condom use behavior using the Transtheoretical Model Stages of Change. Participants drawn from the inactive comparison condition of a randomized HIV prevention trial (Project SHIELD) responded to items assessing pros and cons of condom use, peer norms, condom communication, and perceived invulnerability to HIV. Participants were categorized based on their condom use behavior using the Transtheoretical Model. Multiple logistic regressions found that progression to consistent condom use was predicted by continuing to perceive more advantages to condom use, reporting greater condom use communication with partners, and less perceived invulnerability to HIV. Movement away from adopting consistent condom use was predicted by a decrease in perceived advantages to condom use, increased perceived condom disadvantages, and fewer condom discussions. Future interventions may be tailored to enhance these factors that were found to change over time. PMID- 18427972 TI - Social discrimination, concurrent sexual partnerships, and HIV risk among men who have sex with men in Shanghai, China. AB - We examined the potential mediating role of sex partner concurrency in explaining associations between experiences of homophobia and financial hardship and HIV risk behavior among a sample of 477 men who have sex with men (MSM) recruited in Shanghai, China from 2004 to 2005. We found significant positive direct associations of experiences of homophobia and financial hardship with having unprotected anal sex with men. These associations were not mediated by male partner concurrency. However, male partner concurrency had a direct effect on having unprotected anal sex with men. Male and female partner concurrency did not mediate the significant association between experiences of homophobia and having unprotected anal/vaginal sex with both men and women, but was positively associated with having unprotected anal/vaginal sex with both men and women. These results suggest that homophobia, financial hardship, and sex partner concurrency should be addressed to help reduce sexual risk for HIV among MSM in China. PMID- 18427973 TI - The challenges of assessing fidelity to physician-driven HIV prevention interventions: lessons learned implementing Partnership for Health in a Los Angeles HIV clinic. AB - Documenting fidelity to HIV prevention interventions is critical to ensure consistency in intervention implementation and necessary for measuring intervention exposure and, ultimately, outcomes. Significant variation from prescribed protocols or inconsistent implementation can jeopardize the integrity of evaluation research and render outcomes uninterpretable. There is increasing support for HIV prevention models targeting seropositive individuals designed to be delivered by physicians during clinic visits. Assessing fidelity to physician delivered interventions that occur during clinical exams present unique challenges. This paper presents findings from various data sources designed to track intervention fidelity and exposure to the Partnership for Health intervention, a physician-delivered HIV prevention intervention implemented in an urban community HIV clinic. We present findings from chart abstraction data, patient surveys and exit interviews, and provider qualitative interviews. Lessons learned and recommendations for maximizing the accuracy and validity of fidelity assessment in future evaluations of HIV prevention interventions in primary care settings are considered. PMID- 18427974 TI - Comparison of the efficacy of the aromatase inhibitor letrozole and clomiphen citrate gonadotropins in controlled ovarian hyperstimulation: a prospective, simply randomized, clinical trial. AB - OBJECTIVE: To study the efficacy of the aromatase inhibitor letrozole in controlled ovarian hyperstimulation (COH). MATERIAL AND METHODS: In this prospective simply randomized clinical trial, one hundred forty patients with unexplained infertility undergoing intrauterine insemination (IUI) therapy were randomized to receive either letrozole or clomiphene citrate (CC)-gonadotropin. The patients were selected among patients referred to one university hospital and one private infertility clinic. A letrozole dose of 5 mg/day (n = 70) was given on days 3-7 of the menstrual cycles. Clomiphen citrate a dose of 100 mg/day was given like letrozole but combined with human menopausal gonadotropin (hMG) dose of 75 IU/ml administered every day starting on day 6. Ovulation was triggered with urinary hCG (10,000 IU) when the leading follicle(s) reached 18 mm in diameter. A single IUI was performed 36 hours later. The luteal phase was supplemented with micronized progesterone vaginally. Ovarian stimulation response (E2 levels and number of follicles) was primary outcome. RESULTS: There were no differences in demographic characteristics between groups. The number of mature follicles (1.8 +/- 0.7 vs. 2.46 +/- 2.3; P = 0.042) and serum E2 level on the day of hCG (310 +/- 135.4 vs. 1,670.7 +/- 1021.8 pg/ml, respectively; P < 0.0001) were significantly lower in letrozole group. A significantly higher endometrial thickness was observed at the time of hCG administration in patients that received letrozole (9.7 +/- 1.6 mm vs. 7.8 +/- 2 mm; P < 0.001). Clinical pregnancy rates also were significantly higher in letrozole group (32.8% vs. 14.3%, respectively; P < 0.01). CONCLUSION: The aromatase inhibitor letrozole appears to constitute a good alternative to CC-gonadotropin in patients with unexplained infertility undergoing COH cycles combined with IUI therapy. PMID- 18427975 TI - Do theory of mind and executive function deficits underlie the adverse outcomes associated with profound early deprivation?: findings from the English and Romanian adoptees study. AB - Theory of Mind (ToM) and Executive Function (EF) have been associated with autism and with attention-deficit hyperactivity disorder (ADHD), and hence might play a role in similar syndromes found following profound early institutional deprivation. In order to examine this possibility the current study included a group of 165 Romanian adoptees, of whom 144 were adopted into the UK from deprived institutional settings before 43months of age, and a group of 52 within UK adoptees, all adopted before 6months of age. Both groups were assessed at 6 and 11years. The Strange Stories task was used to assess ToM and the Stroop task was used to assess EF, both at age 11. The Romanian adoptees displayed deficits in both ToM and EF compared with the within-UK adoptee group. The degree of deficit was greater for children who had experienced more than 6months of institutional deprivation. Deficits in both domains (ToM and EF) were associated with each of the three apparently deprivation-specific problems, namely quasi autism, disinhibited attachment and inattention/overactivity. Statistical analyses indicated a mediating role for both ToM and EF with respect to quasi autism; possibly a partial mediating role for EF with respect to inattention/overactivity; and probably no mediating role for either ToM or EF in the case of disinhibited attachment. In conclusion, there is evidence for a possible mediating role for ToM and EF in the development of some apparently deprivation-specific difficulties in institution-reared Romanian adoptees, but neither accounts for the overall pattern of deprivation-related difficulties. PMID- 18427976 TI - Justification of violence and grandiosity schemas as predictors of antisocial behavior in adolescents. AB - This study assessed the role of grandiosity and justification of violence cognitive schemas as predictors of adolescents' antisocial behavior. The 974 Spanish adolescents (457 girls and 517 boys, aged between 14 and 18 years) were assessed at the beginning of the school year and at follow-up 6 months later. They completed measures of aggressive behavior and delinquent behavior, justification of violence and grandiosity cognitive schemas, and impulsivity. The results showed that grandiosity and justification of violence schemas predicted both aggressive and delinquent behavior 6 months later and that the association between grandiosity and aggressive behavior was moderated by impulsivity in boys. In general, the influence of the cognitive schemas in antisocial behavior was very similar for boys and girls. However, boys showed higher delinquent behavior rates than did girls and the tests of mediation revealed that this difference was partially accounted for by boys' higher scores in grandiosity and justification of violence. PMID- 18427977 TI - High-throughput multiplex single-nucleotide polymorphism (SNP) analysis in genes involved in methionine metabolism. AB - Hyperhomocysteinemia is a well-known independent marker factor for atherothrombotic diseases and may result from acquired and genetic influences. Several polymorphisms are suspected to be associated with hyperhomocysteinemia, but data are limited and inconsistent. High-throughput genotyping technologies, such as GenomeLab SNPStream, are now available. Moreover, an appropriate selection of SNPs to be analyzed could represent a strong resource to define the role of genetic risk factors. We developed a multiplex PCR-oligonucleotide extension approach by GenomeLab platform. We selected 72 SNPs based on their putative function and frequency in the candidate genes AHCY, BHMT, BHMT2, CBS, ENOSF1, FOLH1, MTHFD1, MTHFR, MTR, MTRR, NNMT, PON1, PON2, SLC19A1, SHMT1, TCN2, and TYMS. We were able to analyze 57 of the SNPs (79%). For MTHFR C677T and A1298C and MTR A2756G SNPs, we compared data obtained with an electronic microchip technology and found 99.2% concordance. We also performed a haplotype analysis. This approach could represent a useful tool to investigate the genotype phenotype correlation and the association of these genes with hyperhomocysteinemia and correlated diseases. PMID- 18427978 TI - Chromosome location and association of haplotypes of insulin-like growth factor binding protein-2 with production performance in swine. AB - Insulin-like growth factor binding protein-2 is a member of the insulin-like growth factor families. Using a porcine RH panel, the gene was mapped on chromosome 15q22-23. Meanwhile, using polymerase chain reaction single strand conformation polymorphism, genotypic and allelic frequencies were analyzed in 17 pig breeds (total animals 570), together with a chi-square test of Hardy-Weinberg equilibrium. Also the association between haplotypes and production performance was analyzed in a Lantang x Landrace population family (n = 133, total 43 traits). At each locus we investigated, all the breeds showed different genotypic and allelic frequency distributions. In general, the Chinese native pig breeds carried a higher allele A frequency (over 50%) than the European pigs. For production performance, pigs with the CAG haplotype had higher fore-body and rear body weight than those with the TGT and TAG haplotypes (P < 0.05). Also, pigs with the CAG haplotype had higher bone weight of the rear-body than those with the CAT haplotype (P < 0.05); pigs with the TGT and CAG haplotypes had higher forelimb and rearlimb weight than those with the CAT haplotype (P < 0.01 and P < 0.05, respectively); pigs with the TGG haplotype had higher leaf fat weight than those with the TGT and CAG haplotypes (P < 0.05); and pigs with the CAG haplotype had more stomach weight than those with the CAT and CGT haplotypes (P < 0.01); pigs with the TGT and CAG haplotypes had more ribs and longer body than those with the CGT-TGG, and CAT-TAG haplotypes (P < 0.05). These results suggest that IGFBP-2 is associated with production performance, but our population family was small. More studies with large samples are needed before the IGFBP-2 locus will be useful for a selection program. PMID- 18427979 TI - Effect of Coenzyme Q(10), Riboflavin and Niacin on Tamoxifen treated postmenopausal breast cancer women with special reference to blood chemistry profiles. AB - BACKGROUND: Tamoxifen (TAM) a non-steroidal antiestrogen, is widely used in adjuvant therapy for all stages of breast carcinomas and in chemoprevention of high-risk group. TAM also has estrogenic activity on liver and endometrium causing severe oxidative stress with various biochemical derangements. Coenzyme Q(10), Riboflavin and Niacin (CoRN) are well-known potent antioxidants and protective agents against many diseases including cancer. In this context, this study was undertaken to find if co-administration of TAM along with CoRN could alleviate the sole TAM-induced biochemical derangements in postmenopausal women with breast cancer. METHOD: The vitamin supplementation with TAM was given for a period of 90 days. Blood samples were collected at the base line, 45th and 90th day during the course of treatment. Various blood chemistry profiles were assessed in 78 untreated, sole TAM treated and combinatorial treated group along with 46 age- and sex-matched controls. RESULTS: A statistically significant alteration in various blood chemistry parameters, such as serum total bilirubin (S. BIL), serum glutamate oxaloacetate transaminase (SGOT), serum glutamate pyruvate transaminase (SGPT), gamma glutamyl transpeptidase (gamma-GT), uric acid (UA), lipoprotein lipase (LPL), lecithin: cholesterol acyl transferases (LCAT), potassium, calcium and Na(+), K(+)-ATPase in sole TAM-treated group, was favorably reverted back to near normal levels on combinatorial therapy with CoRN. CONCLUSION: TAM on co-administration with CoRN has a favorable impact on various blood chemistry profiles. However, large scale randomized studies over a longer time span are required to ascertain the safety and efficacy of co-administrating antioxidants with conventional chemotherapy. PMID- 18427981 TI - The Wnt antagonist, Dickkopf-1, as a target for the treatment of neurodegenerative disorders. AB - The canonical Wnt pathway contributes to the regulation of neuronal survival and homeostasis in the CNS. Recent evidence suggests that an increased expression of Dickkopf-1 (Dkk-1), a secreted protein that negatively modulates the canonical Wnt pathway, is causally related to processes of neurodegeneration in a number of CNS disorders, including Alzheimer's disease (AD), brain ischemia and temporal lobe epilepsy (TLE). Dkk-1 induction precedes neuronal death in cellular and animal models of excitotoxicity, beta-amyloid toxicity, transient global ischemia, and kainate-induced epilepsy. In addition, Dkk-1, which is barely visible in the healthy brain, is strongly induced in brain tissue from AD patients or from patients with TLE associated with hippocampal sclerosis. These data raise the attractive possibility that Dkk-1 antagonists or neutralizing antibodies behave as neuroprotective agents by rescuing the activity of the canonical Wnt pathway. PMID- 18427982 TI - The use of CDME in cystinosis research. PMID- 18427980 TI - Intracellular signaling in primary sensory neurons and persistent pain. AB - During evolution, living organisms develop a specialized apparatus called nociceptors to sense their environment and avoid hazardous situations. Intense stimulation of high threshold C- and Adelta-fibers of nociceptive primary sensory neurons will elicit pain, which is acute and protective under normal conditions. A further evolution of the early pain system results in the development of nociceptor sensitization under injury or disease conditions, leading to enhanced pain states. This sensitization in the peripheral nervous system is also called peripheral sensitization, as compared to its counterpart, central sensitization. Inflammatory mediators such as proinflammatory cytokines (TNF-alpha, IL-1beta), PGE(2), bradykinin, and NGF increase the sensitivity and excitability of nociceptors by enhancing the activity of pronociceptive receptors and ion channels (e.g., TRPV1 and Na(v)1.8). We will review the evidence demonstrating that activation of multiple intracellular signal pathways such as MAPK pathways in primary sensory neurons results in the induction and maintenance of peripheral sensitization and produces persistent pain. Targeting the critical signaling pathways in the periphery will tackle pain at the source. PMID- 18427984 TI - WITHDRAWN: Synthetic NCAM-derived Ligands of the Fibroblast Growth Factor Receptor. PMID- 18427983 TI - Epileptic seizures but not pseudoseizures are associated with decreased density of the serotonin transporter in blood platelet membranes. AB - The density of the serotonin transporter in the plasma membranes of blood platelets was evaluated by labelled paroxetine binding in three different groups. These groups were: normal controls, epileptic patients having undergone a recent seizure (less than 4 days before) and patients who equally recently presented psychogenic non-epileptic seizures (pseudoseizures). Real seizures resulted in a significant decrease of membrane serotonin transporter density. In the instances of pseudoseizures, its membrane density was undistinguishable from that of normal controls. These data lend further support to the idea that down regulation of serotonin transporter may play a homeostatic role in the cessation of epileptic seizures. PMID- 18427985 TI - An analytical flow injection system to measure glutamate in microdialysis samples based on an enzymatic reaction and electrochemical detection. AB - Glutamate (Glu) is the main excitatory neurotransmitter in the brain for which several methods have been developed to measure this compound in extracellular brain fluids. Most of these techniques are based on coupling microdialysis to HPLC and they have a resolution time of about 10 min. Here, we present a different approach to measure Glu with a resolution of about 1 min per microdialysis sample, enabling a better relationship to be established between EEG activity and biochemical changes. This new setup was used to determine the time delay between the tip of the microdialysis probe and the site of sample collection, and was accurate to within seconds. Indeed, the measurement of Glu concentrations was linear. Administration of 4-aminopyridine was used to provoke seizure convulsions and under these conditions, biochemical changes and EEG activity were evaluated. These experimental data support the key role of Glu in the initiation of a seizure convulsion. PMID- 18427986 TI - Protective effects of Nigella sativa on the neuronal injury in frontal cortex and brain stem after chronic toluene exposure. AB - The goal of this study was designed to evaluate the possible protective effects of Nigella sativa (NS) on the neuronal injury in the frontal cortex and brain stem after chronic toluene exposure in rats. The rats were randomly alotted into one of three experimental groups: A (control), B (toluene treated) and C (toluene treated with NS); each group contain 10 animals. Control group received 1 ml serum physiologic and toluene treatment was performed by inhalation of 3,000 ppm toluene, in a 8 h/day and 6 day/week order for 12 weeks. The rats in NS treated group was given NS (in a dose of 400 mg/kg body weight) once a day orally by using intra gastric intubation for 12 weeks starting just after toluene exposure. Tissue samples were obtained for histopathological investigation. To date, no histopathological changes of neurodegeneration in the frontal cortex and brain stem after chronic toluene exposure in rats by NS treatment have been reported. In this study, chronic toluene exposure caused severe degenerative changes, shrunken cytoplasma, severely dilated cisternae of endoplasmic reticulum, markedly swollen mitochondria with degenerated cristae and nuclear membrane breakdown with chromatin disorganization in neurons of the frontal cortex and brain stem. The nerve cells showing the pathologic changes were almost absent in the NS-treated rats. We conclude that NS therapy causes morphologic improvement on neurodegeneration in frontal cortex and brain stem after chronic toluene exposure in rats. We believe that further preclinical research into the utility of NS may indicate its usefulness as a potential treatment on neurodegeneration after chronic toluene exposure in rats. PMID- 18427987 TI - Parvalbumin immunoreactivity and protein level are altered in the gerbil hippocampus during normal aging. AB - Hippocampal interneurons are local circuit neurons which are responsible for inhibitory activity in the hippocampus. Parvalbumin (PV) is one of useful markers for GABAergic interneurons, not for principle cells, in the hippocampus. In the present study, we investigated age-related changes in PV immunoreactive neurons and protein levels in the gerbil hippocampus during normal aging. PV immunoreactive neurons were detected in all hippocampal subregions of all groups. PV immunoreactive neurons, which innervated principal neurons, were non-pyramidal neurons in the hippocampal CA1-3 regions, and were polymorphic neurons in the dentate gyrus. In the hippocampal CA1 region, the number of PV immunoreactive neurons was significantly reduced in the postnatal month 3 (PM 3) group, which was sustained by PM 18, and, at PM 24, the number of PV immunoreactive neurons was significantly decreased. In the CA2/3 region and dentate gyrus, the number of PV immunoreactive neurons was significantly decreased at PM 6: Thereafter, the number of PV immunoreactive neurons was sustained until PM 24. In addition, changes in PV protein levels in the gerbil hippocampus were similar to immunohistochemical changes during normal aging: PV protein levels were significantly decreased with age by PM 6: Thereafter, PV protein levels were sustained by PM 24. These results suggest that PV immunoreactive interneurons were decreased in the hippocampus with age in gerbils. PMID- 18427988 TI - The PKC and ERK/MAPK pathways regulate glucocorticoid action on TRH transcription. AB - Biosynthesis of TRH, a neuropeptide involved in energy homeostasis, is modulated by glucocorticoids. TRH mRNA and peptide levels are increased upon incubation of hypothalamic cells with dexamethasone or with cAMP analogs but when combined, a mutual antagonism is observed. These effects are observed at the transcriptional level and on binding of glucocorticoid receptor (GR) or pCREB to the composite GRE (cGRE) and CRE-2 sites of TRH promoter. The present work studied the involvement of PKC and MAPK pathways on the effect of dexamethasone and on its interaction with cAMP signaling in hypothalamic cell cultures. PKC or MEK inhibition abolished dexamethasone-stimulatory effect on TRH mRNA levels, as well as its interference with the stimulatory effect of 8Br-cAMP. Binding of nuclear extracts from hypothalamic or neuroblastoma cells stimulated with dexamethasone or 8Br-cAMP to oligonucleotides containing the CRE or cGRE sites of TRH gene promoter was decreased if cells were preincubated with PKC or MEK inhibitors. Mutations on the AP-1 or the GRE half sites of cGRE showed that GR binds as an heterodimer on cGRE, and PKC or MEK inhibitors diminish binding at the AP-1 site. PKC and ERK signaling thus modulate GR activity and its interaction with CREB or AP-1 at the TRH gene promoter. PMID- 18427989 TI - Effects of naltrexone and acamprosate on alcohol-induced NGFI-A expression in mouse brain. AB - In search for the substrate of naltrexone and acamprosate action on alcohol craving, we investigated the effects of ethanol alone and combined with naltrexone or acamprosate on expression of nerve growth factor-inducible clone A (NGFI-A; zif268). In Experiments 1 and 3, alcohol (2 g/kg) alone or in combination with naltrexone (15 mg/kg) or acamprosate (300 mg/kg) was injected intraperitoneally into mice. In Experiment 2, treatment was nor-BNI (0.5 mg/kg) to investigate whether the effect of naltrexone involved blockade of kappa-opioid receptors. Both ethanol and naltrexone alone induced NGFI-A in the central amygdala, but not in several other areas; these effects were additive. However, acamprosate alone or in combination with ethanol had no effect on NGFI-A mRNA, while nor-BNI induced NGFI-A mRNA in the basolateral amygdala. The central amygdala appears to be an important target of both alcohol and naltrexone. Acamprosate may not share the site of action with naltrexone despite being used for the same therapeutic purpose. PMID- 18427990 TI - Yoghurts containing probiotics reduce disruption of the small intestinal barrier in methotrexate-treated rats. AB - Small intestinal permeability was employed to assess the efficacy of commercially available yoghurts containing probiotics in a rat model of methotrexate (MTX) induced mucositis. Male Sprague-Dawley rats were allocated to four groups (n = 8): MTX + water, MTX + cow's milk yoghurt (CY; fermented with Lactobacillus johnsonii), MTX + sheep's milk yoghurt (SY; containing Lactobacillus bulgaricus and Streptococcus thermophilus), and saline. Treatment gavage occurred twice daily for 7 days pre-MTX and 5 days post-MTX. Intestinal permeability was assessed on days -7, -1, 2, and 5 of the trial. Intestinal sections were collected at sacrifice for histological and biochemical analyses. Histology revealed that rats receiving CY and SY did not have a significantly damaged duodenum compared to controls. However, an improved small intestinal barrier function was evident, determined by a decreased lactulose/mannitol ratio. Probiotics containing SY and CY may be useful in preventing disruption to intestinal barrier function in MTX-induced mucositis. PMID- 18427991 TI - Does cyclophosphamide play a protective role against neuronal loss in chronic T. cruzi infection? AB - In this study, we verified the possible role of cyclophosphamide (CY) in protecting or not against neuronal losses in young and aged male Calomys callosus chronically infected with the MORC-1 strain of Trypanosoma cruzi through numerical quantification of neurons from the myenteric plexus of the colon and quantification of nitric-oxide concentration (NO) during the acute and chronic phase of infection. For this purpose, groups of young C. callosus were infected with the MORC-1 strain of T. cruzi. A group of infected animals received i.p. 0.2 mg/ml genuxal dissolved in distilled water treatment with CY. NO concentration in aged animals displayed reduced levels when compared to those found in young animals. No significant alterations in the number of neurons were observed in young animals, but for aged ones, a protective role of CY in reducing neuron loss was noted, in addition to enhancing the neuronal volume, area, and perimeter. These results suggest that CY administration, depending on the dose and time span, can act as a protective agent against neuronal losses. PMID- 18427992 TI - A systematic review and meta-analysis of the treatment for Barrett's esophagus. AB - As evidence-based strategies to the clinical management of Barrett's esophagus (BE) are lacking, we have carried out a systematic review and meta-analysis of all published randomized controlled trials with the aim of evaluating the value of different approaches in the treatment of BE. Searches were conducted in the databases PUBMED, EMBASE, and Cochrane Library. Thirteen randomized clinical trials that fulfilled the inclusion criteria and addressed the clinical questions of this analysis were assessed in more detail. Based on our search, neither the pharmacological nor surgical therapies currently available for reflux appear to achieve complete regression of BE and the elimination of the cancer risk associated with it. In contrast, endoscopic ablative techniques are capable of achieving endoscopic and histological reversal of BE, with ablation by argon plasma coagulation (APC) appearing to be more effective than treatment with photodynamic therapy (PDT) [odds ratio (OR) 3.46, 95% confidence interval (CI) 1.67-7.81, P = 0.0008]. There was no statistically significant difference between APC and multipolar electrocoagulation (MPEC) in terms of the efficacy to achieve regression of BE (OR 2.01, 95% CI 0.77-5.23, P = 0.15). In conclusion, there have been only a limited number of randomized controlled trials that compare treatments for BE. The pharmacological therapy, antireflux surgery, and endoscopic ablative techniques are promising in terms of treating BE, but the studies carried out to date have had no adequate power to assess the effect of treatment on reducing and preventing progression to adenocarcinoma. PMID- 18427993 TI - An investigation of vibration-induced protein desorption mechanism using a micromachined membrane and PZT plate. AB - A micromachined vibrating membrane is used to remove adsorbed proteins on a surface. A lead zirconate titanate (PZT) composite (3 x 1 x 0.5 mm) is attached to a silicon membrane (2,000 x 500 x 3 microm) and vibrates in a flexural plate wave (FPW) mode with wavelength of 4,000/3 microm at a resonant frequency of 308 kHz. The surface charge on the membrane and fluid shear stress contribute in minimizing the protein adsorption on the SiO(2) surface. In vitro characterization shows that 57 +/- 10% of the adsorbed bovine serum albumin (BSA), 47 +/- 13% of the immunoglobulin G (IgG), and 55.3~59.2 +/- 8% of the proteins from blood plasma are effectively removed from the vibrating surface. A simulation study of the vibration-frequency spectrum and vibrating amplitude distribution matches well with the experimental data. Potentially, a microelectromechanical system (MEMS)-based vibrating membrane could be the tool to minimize biofouling of in vivo MEMS devices. PMID- 18427995 TI - A new missense mutation found in the FLNA gene in a family with bilateral periventricular nodular heterotopia (BPNH) alters the splicing process. AB - We describe the clinical and molecular evaluation of two patients, mother and daughter (proband), with bilateral periventricular nodular heterotopia (BPNH). The clinical evaluation revealed a more severe phenotype in the proband, with mental retardation and seizures. Imaging studies showed bilateral periventricular nodules in both patients. We identified a novel mutation, c.987G-->C mutation in exon 6 of the Filamin A (FLNA) gene in the genomic DNA of both patients. Complementary DNA (cDNA) sequencing revealed the maintenance of intron 6 in the mutated allele. Bioinformatics analysis indicates that the mutation identified in both patients probably destroyed the intron 6 donor-splicing site, which is likely to introduce a premature stop codon resulting in a truncated FLNA protein. In addition, X-chromosome inactivation studies in DNA of blood cells revealed a skewed pattern in the proband, and real time quantitative polymerase chain reaction (PCR) showed a higher expression of the mutated allele in the proband compared to that of the mother. This variation in expression of the mutated allele may be responsible for the differences in the clinical manifestations observed in both patients. PMID- 18427996 TI - Gene manipulation of a heavy metal hyperaccumulator species Thlaspi caerulescens L. via Agrobacterium-mediated transformation. AB - Thlaspi caerulescens L. is well known as a Zn/Cd hyperaccumulator. The genetic manipulation of T. caerulescens through transgenic technology can modify plant features for use in phytoremediation. Here, we describe the efficient transformation of T. caerulescens using Agrobacterium tumefaciens strain EHA105 harboring a binary vector pBI121 with the nptII gene as a selectable marker, the gus gene as a reporter and a foreign catalase gene. Based on the optimal concentration of growth regulators, the shoot cluster regeneration system via callus phase provided the basis of the genetic transformation in T. caerulescens. The key variables in transformation were examined, such as co-cultivation period and bacterial suspension density. Optimizing factors for T-DNA delivery resulted in kanamycin-resistant transgenic shoots with transformation efficiency more than 20%, proven by histochemical GUS assay and PCR analysis. Southern analysis of nptII and RT-PCR of catalase gene demonstrated that the foreign genes were integrated in the genome of transformed plantlets. Moreover, the activity of catalase enzyme in transgenic plants was obviously higher than in wild-type plants. This method offers new prospects for the genetic engineering of this important hyperaccumulator species. PMID- 18427997 TI - [Preliminary study aimed at the construction of a questionnaire for referral from primary care to mental health services]. AB - The high prevalence of psychiatric morbidity in primary care, the growing perception of the need for specialized help by the least severe patients and the lack of accuracy in referrals, contribute to the increasing overload in mental health services. So it seems necessary to design diagnostic tools in order to improve the detection of more severe patients and to help in the referral decision. With this purpose in mind, we have designed the multidimensional heteroadministrated Scale Referral Criteria for Mental Health (CRMH). This paper presents the preliminary results of a pilot study on its application in a sample of 198 patients by a group of Primary Care Physicians (PCP). The data show the detection of a high percentage of potential psychiatric patients (46.9%) and 4% of patients who having the possibility of being referred. The results also illustrate the low ability of PCP to detect these disorders. CRMH has a moderate correlation with the General Health Questionnaire (GHQ) and with detection of psychopathology by PCP. Other factors, apart from clinical severity evaluated through CRMH, possibly belonging to doctor-patient relationship, which should be analyzed, seem to influence the mental health referral. In a future article, we will present the validation of this scale in our care setting. PMID- 18427998 TI - Behaviour in amyotrophic lateral sclerosis. AB - We wished to determine whether patients with amyotrophic lateral sclerosis (ALS) show behavioural changes similar to those of frontotemporal dementia (FTD). There is accumulating evidence to suggest a link between ALS and FTD, yet there has been little systematic exploration of behaviour in ALS. An informant based semi structured behavioural interview, sensitive to the behavioural changes of FTD, was administered to carers of 16 consecutive patients attending a motor neuron disease clinic. Findings varied across the group. At one extreme informants reported no behavioural change, whereas at the other they reported a spectrum of behaviours similar to those seen in FTD. Changes in affect and social behaviour were most common, although some patients also showed altered response to sensory stimuli, gluttony and indiscriminate eating, behavioural stereotypies and compulsions. Behavioural changes were mirrored by SPECT abnormalities in the frontal and/or temporal lobes. Thus, behavioural changes of the type seen in FTD may be present even in a small consecutive cohort of ALS patients. Detection of behavioural change is crucial for optimal management. PMID- 18427999 TI - Association study on glutathione S-transferase omega 1 and 2 and familial ALS. AB - Glutathione S-transferase omega 1 and 2 (GSTO1 and 2) protect from oxidative stress, a possible pathogenic mechanism underlying the pathogenesis of neurodegenerative diseases such as amyotrophic lateral sclerosis (ALS) and Alzheimer's disease. Significant association of age of onset in Alzheimer's patients with GSTO1 and 2 had recently been identified, suggesting a possibly similar association with ALS. In this study 12 Hapmap tagged SNPs in GSTO1 and 2 were genotyped in 251 Caucasian British, Australian and Swedish familial ALS (FALS) cases. No association was found for age of onset and survival of FALS in the British and Australian patients. In the Swedish patients, association for age of onset was found with several SNPs (p = 0.003-0.048). These results suggest a possible effect of the GSTO1 and 2 locus on age of onset of FALS. PMID- 18428000 TI - Gemals, a new drug candidate, extends lifespan and improves electromyographic parameters in a rat model of amyotrophic lateral sclerosis. AB - Amyotrophic lateral sclerosis (ALS) is a fatal disease involving selective and progressive degeneration and death of motor neurons. ALS is a multifactorial disease in which oxidative stress, glutamate excitotoxicity, intracellular aggregates, neurofilamentous disorganization, zinc excitotoxicity, mitochondrial damage, neuroinflammation, abnormalities in growth factors and apoptosis play a role. Any therapeutic approach to delay or stop the evolution of ALS should therefore ideally target these multiple pathways leading to motor neuron death. We have developed a combination therapy (Gemals) composed of functional polypeptides (fatty acids, free radical scavengers and amino acids linked to poly L-lysine), chosen according to their known potentiality for regeneration or protection of neuronal components such as myelin, axon transport and mitochondria. We found that Gemals significantly extended lifespan and improved electromyographic parameters in a SOD1(G93A) rat model. The use of two drug concentrations indicated a possible dose dependence. These initial findings open the way to further investigation necessary to validate this new drug as a candidate for ALS treatment. PMID- 18428001 TI - Palliative care and circumstances of dying in German ALS patients using non invasive ventilation. AB - Non-invasive ventilation (NIV) is known to improve quality of life and to prolong survival in amyotrophic lateral sclerosis (ALS) patients. However, little is known about the circumstances of dying in ventilated ALS patients. In the light of the debate on legalizing euthanasia it is important to provide empirical data about the process of dying in these patients. In a structured interview, 29 family caregivers of deceased ALS patients were asked about their own and the patient's attitude toward physician-assisted suicide (PAS) and euthanasia, circumstances of dying, and the use of palliative medication. Quantitative and qualitative content analysis was performed on the data. Non-recurring suicidal thoughts were reported by five patients. Three patients and seven relatives had thought about PAS. Seventeen caregivers described the patients' death as "peaceful", while choking was reported in six bulbar patients. In final stages of dying, the general practitioner (GP) was involved in the treatment of 10 patients, with palliative medication including sedatives and opiates being administered in eight cases. In conclusion, in contrast to the Netherlands, where 20% of terminal ALS patients die from PAS or euthanasia, only a small minority of our patients seems to have thought about PAS. The legal situation in Germany (where euthanasia is illegal), a bias due to the selection of NIV patients as well as a high percentage of religious patients and those with good levels of social support from family and friends, might account for this. Most of our patients died peacefully at home from carbon dioxide narcosis, but choking was described in some bulbar patients. Thus, palliative care, especially the use of opiates, anxiolytics and sedatives should be optimized, and the involvement of GP should be strongly encouraged, especially in bulbar patients. PMID- 18428002 TI - The influence of personality factors on disease progression and health-related quality of life in people with ALS. AB - We aimed to perform a prospective long-term follow-up of health-related quality of life (QOL) in ALS and to investigate the relationship of personality factors with changes in QOL and disease progression. Data on QOL were collected prospectively for 12 months from 31 ALS patients. Personality factors were studied using the NEO-FFI (NEO Five Factor Inventory). Monthly self-ratings of global QOL, and seven health-related QOL functions, as well as ALSFRS (ALS Functional Rating Scale) scores were analyzed using a linear mixed model approach. QOL and ALSFRS scores decreased during follow-up. Patients who scored higher on the agreeableness personality dimension, despite similar total duration of disease, had higher QOL at the beginning of the follow-up period but the reduction of QOL over time was significantly steeper than in patients who scored lower on agreeableness, associated with faster disease progression. These findings suggest that being less agreeable might serve as a protective factor with respect to QOL and disease progression in ALS. PMID- 18428003 TI - SOD1 gene mutations in ALS patients from British Columbia, Canada: clinical features, neurophysiology and ethical issues in management. AB - Two hundred and fifty-four ALS patients from British Columbia, Canada were screened for mutations in the gene encoding the enzyme superoxide dismutase type 1 (SOD1). Thirteen patients (5.1%) carried one of six missense mutations (A4V, G72C, D76Y, D90A, C111Y, I113T). Mutations were found both in sporadic and familial ALS cases. Atypical clinical features delayed diagnosis in some cases. The demographic and clinical features of the mutation carrying index cases are summarized, and compared with those of screened patients without mutations. The phenotypic variability between SOD1 mutation carrying patients in this study is dramatic, even among patients with the same mutation This underlines the hypothesis that ALS is a biologically heterogeneous disorder in which genetics, environment and ageing all interrelate to form the final clinical phenotype. PMID- 18428004 TI - Strength, physical activity, and fasciculations in patients with ALS. AB - Fasciculations are a nearly universal feature in people with amyotrophic lateral sclerosis (ALS). The prognostic value of fasciculations remains uncertain. Twenty four patients with ALS were evaluated for the effects of atrophy, limb weakness, disease duration, and physical activity on fasciculation frequency (as measured by surface electromyography and clinical counting). Variables were compared by multiple linear regression. As strength of the limb deteriorated, the number of fasciculations in the same limb increased, as long as physical activity was maintained or increased. Fasciculation frequency was not associated with the duration of ALS (r = 0.22; p = 0.30) and was independent of the degree of limb weakness (p>0.05) and limb atrophy (p>0.05). No prediction of disease duration could be made based on fasciculation frequency alone. Fasciculations therefore appear to have diagnostic, but not prognostic, utility in the care of people with ALS. PMID- 18428005 TI - Lithium therapy in ALS. PMID- 18428006 TI - Lithium in ALS: from the bench to the bedside. PMID- 18428007 TI - Comments on Fornai et al. (PNAS/ Feb 2008). PMID- 18428008 TI - Ethnogenetic layering (EL): an alternative to the traditional race model in human variation and health disparity studies. AB - BACKGROUND: Traditionally, studies in human biodiversity, disease risk, and health disparities have defined populations in the context of typological racial models. However, such racial models are often imprecise generalizations that fail to capture important local patterns of human biodiversity. AIM: More explicit, detailed, and integrated information on relevant geographic, environmental, cultural, genetic, historical, and demographic variables are needed to understand local group expressions of disease inequities. This paper details the methods used in ethnogenetic layering (EL), a non-typological alternative to the current reliance of the biological racial paradigm in public health, epidemiology, and biomedicine. SUBJECTS AND METHODS: EL is focused on geographically identified microethnic groups or MEGs, a more nuanced and sensitive level of analysis than race. Using the MEG level of analysis, EL reveals clinical variations, details the causes of health disparities, and provides a foundation for bioculturally effective intervention strategies. EL relies on computational approaches by using GIS-facilitated maps to produce horizontally stratified geographical regional profiles which are then stacked and evaluated vertically. Each horizontal digital map details local geographic variation in the attributes of a particular database; usually this includes data on local historical demography, genetic diversity, cultural patterns, and specific chronic disease risks (e.g. dietary and toxicological exposures). Horizontal visual display of these layered maps permits vertical analysis at various geographic hot spots. RESULTS AND CONCLUSIONS: From these analyses, geographical areas and their associated MEGs with highly correlated chronic disease risk factors can be identified and targeted for further study. PMID- 18428009 TI - Growth and nutritional status in children and adolescents with cystic fibrosis. AB - BACKGROUND: Growth retardation, delayed puberty and malnutrition are frequently observed in children suffering from cystic fibrosis. AIM: The aim of this study was to estimate growth and nutritional status in children with cystic fibrosis on the basis of body proportions and body mass index. SUBJECTS AND METHODS: Anthropometric data were collected from the medical histories of 62 patients treated in three cystic fibrosis treatment centers in Poland. Anthropometric parameters were expressed in terms of standard deviations away from age-specific and sex-specific reference means reported for the population of Poland. Two-way analysis of variance was used to determine whether the type of cystic fibrosis transmembrane conductance regulator (CFTR) mutation is correlated with age at the time of diagnosis and with body proportions. RESULTS: The type of mutation was significantly correlated with height, weight and transverse chest width. Growth retardation was greater in subjects diagnosed before they were 3 years old than in subjects diagnosed later. The children had infantile body proportions. Their legs were short and their trunks were long in comparison to their height. Almost 40% of the subjects suffered from malnourishment. CONCLUSION: Further study is needed to determine how growth in children with cystic fibrosis is affected by clinical practice and socio-economic factors. PMID- 18428010 TI - UK adults exhibit higher step counts in summer compared to winter months. AB - BACKGROUND: Seasonal differences in step counts have been observed in a limited number of studies conducted on US adults. Due to the diverse global climate, assessment and interpretation of seasonal patterns in ambulatory activity may vary between countries, and regionally specific studies are necessary to understand global patterns. Currently, no studies have assessed whether a seasonal trend is present when ambulatory activity is measured objectively in adults living in the UK. AIM: The present study investigated whether pedometer determined step counts of adults living in the UK vary between summer and winter. SUBJECTS AND METHODS: Ninety-six adults (52% male, age = 41.0 +/- 12.3 years, BMI = 26.1 +/- 5.1 kg m(-2)) completed a within-subject bi-seasonal pedometer study. All participants completed two 4-week monitoring periods; one during the summer and one the following winter. The same Yamax SW-200 pedometer was worn throughout waking hours during both seasons, and daily step counts were recorded in an activity log. Intra-individual seasonal changes in mean daily steps were analysed using a paired samples t-test. RESULTS: Summer mean daily step counts (10 417 +/- 3055 steps day(-1)) were significantly higher than those reported during the winter (9132 +/- 2841 steps day(-1)) (p < 0.001). A follow-up study conducted the subsequent summer in a sub-sample (n = 28) reinforced this trend. Summer step counts were significantly higher than winter step counts on all days of the week (p < or = 0.001). A significant day of the week effect was present in both seasons, with step counts reported on a Sunday being on average 1500 steps day( 1) lower than those reported Monday through to Saturday. CONCLUSION: Step counts in the sample of UK adults surveyed decreased significantly in the winter compared to the summer, suggesting future pedometer surveillance studies should capture step counts throughout the year for a non-biased reflection of habitual ambulatory activity. Public health initiatives should target these seasonal differences, and opportunities should be provided that encourage individuals to increase their activity levels during the colder, darker months of the year. PMID- 18428011 TI - Estimating body surface area from mass and height: theory and the formula of Du Bois and Du Bois. AB - BACKGROUND: Body surface areas are usually estimated by means of a formula due in its general form to Du Bois and Du Bois (1916), i.e. area = C x mass(a) x height(b), where C, a and b are empirical constants. Its physical basis is unknown. AIM: The present study aimed to explain this formula, correct some errors in the associated literature and provide a clear basis for future developments. SUBJECTS AND METHODS: Use is made of published data, but arguments are largely based on mathematics and modelling. RESULTS: A more fundamental formula is as follows: area = alpha(mass x height)(1/2) + beta(mass/height), where alpha and beta are constants. For realistic values of mass and height the two equations are numerically equivalent. For individuals, beta cannot be negative and b cannot exceed a, but, as regression parameters, these conditions may not be satisfied. This could be due to systematic or statistical relationships between individual values of alpha or beta and the ratio height(3)/mass. Values of alpha, beta, C, a and b are calculated for some published data. CONCLUSIONS: The original type of formula suffices for practical purposes, but the new one is better in analytical contexts when other terms, e.g. for body shape, are to be incorporated. PMID- 18428012 TI - Comparison of field methods to estimate fat mass in children. AB - BACKGROUND: Reliable field methods to measure fat mass (FM) in children may contribute to primary prevention of childhood obesity. AIM: The objective was to compare the accuracy of existing field methods (skinfold thickness (SF), leg-to leg bioelectrical impedance analysis (BIA), anthropometrics for FM measurement in prepubertal European children. SUBJECTS AND METHODS: Reference FM was measured in 55 French children (30 boys, 25 girls; mean age 8.7 years) using a three compartment model: body volume (BV) was assessed by air displacement plethysmography (ADP) and total body water (TBW) was assessed by deuterium dilution. Agreement between field methods and the reference method was assessed using Bland-Altman analyses. Since field methods for FM measurement are reported to be population-dependent, adjustment to the study population was performed using stepwise multiple linear regressions modelling. RESULTS: Even after adjustment, field methods exhibited a high correlation (R(2) = 0.71-0.84) but a moderate agreement (+/-3.32 to +/-4.47 kg for fat mass) with the reference model. Methods based on BIA or SF performed slightly better than those based on anthropometry. CONCLUSIONS: Field methods for FM measurement may be recommended for epidemiological applications, but not for individual follow-up. New field equipment is required to improve accuracy of FM measurement in children and make individual follow-up possible. PMID- 18428013 TI - Origin and evolution of two Yugur sub-clans in Northwest China: a case study in paternal genetic landscape. AB - BACKGROUND: Yugur is an ethnic group that was officially identified by the Chinese Government in 1953. Within the population there are two sub-clans distinctly identified as the Eastern Yugur and Western Yugur, partly because they have different local languages. AIM: A parentage comparison was conducted between the two sub-clans to investigate their genetic relationship. SUBJECTS AND METHODS: Male subjects were chosen from the two clans to investigate their paternal genetic landscape through typing 14 single nucleotide polymorphisms (SNP) and 12 short tandem repeats (STR) of the Y chromosome. RESULTS: Significant differences were revealed between the sub-clans at the haplogroup level. Genetic divergence was also observed by analyses of multidimensional scaling (MDS) and principal components (PC). Genetically, the Eastern Yugur are closer to the Han Chinese and Mongolian people than the Western Yugur. The Uygur people, who share a common ancestor (ancient Huihu) with the Yugur, were genetically separate from both sub-clans of Yugur. Moreover, the constructed phylogenetic network for haplogroup O provided further evidence that the two Yugur sub-groups present an underlying genetic difference. CONCLUSION: Overall, the diffusion of Mongolians during the Mongol Period has affected the Eastern Yugur more than the Western Yugur. The genetic contribution of the Han people to the Eastern Yugur seems to be more pronounced than to the Western Yugur. Besides the two different contributions referred to above, small population size and genetic drift have resulted in the genetic differentiation of the current sub-clans of Yugur. PMID- 18428014 TI - Mitochondrial DNA variation in Jordanians and their genetic relationship to other Middle East populations. AB - BACKGROUND: The Levant is a crucial region in understanding human migrations between Africa and Eurasia. Although some mitochondrial DNA (mtDNA) studies have been carried out in this region, they have not included the Jordan area. This paper deals with the mtDNA composition of two Jordan populations. AIM: The main objectives of this article are: first, to report mtDNA sequences of an urban and an isolate sample from Jordan and, second, to compare them with each other and with other nearby populations. SUBJECTS AND METHODS: The analyses are based on HVSI and HVSII mtDNA sequences and diagnostic RFLPs to unequivocally classify into haplogroups 101 Amman and 44 Dead Sea unrelated individuals from Jordan. RESULTS: Statistical analysis revealed that, whereas the sample from Amman did not significantly differ from their Levantine neighbours, the Dead Sea sample clearly behaved as a genetic outlier in the region. Its outstanding Eurasian haplogroup U3 frequency (39%) and its south-Saharan Africa lineages (19%) are the highest in the Middle East. On the contrary, the lack ((preHV)1) or comparatively low frequency (J and T) of Neolithic lineages is also striking. Although strong drift by geographic isolation could explain the anomalous mtDNA pool of the Dead Sea sample, the fact that its mtDNA lineage composition mirrors, in geographic origin and haplogroup frequencies, its Y-chromosome pool, points to founder effect as the main cause. Ancestral M1 lineages detected in Jordan that have affinities with those recently found in Northwest but not East Africa question the African origin of the M1 haplogroup. CONCLUSION: Results are in agreement with an old human settlement in the Jordan region. However, in spite of the attested migratory spreads, genetically divergent populations, such as that of the Dead Sea, still exist in the area. PMID- 18428016 TI - Hemoglobin variants, hematological parameters and beta-globin gene cluster haplotypes in an isolated Amerindian group from the Orinoco River Delta. AB - BACKGROUND: Several previous studies reported that the Venezuelan Warao Indians presented unusual genetic characteristics. AIM: The present study checked previous reports of a high frequency of hereditary persistence of fetal hemoglobin (HPFH) and examined other hematological traits. SUBJECTS AND METHODS: Standard hematology, electrophoresis on cellulose acetate, fetal hemoglobin alkali denaturation, gamma-globin chain, DNA amplification and sequencing, and denaturing gradient gel electrophoresis determinations were performed in 269 individuals living in two localities of the Orinoco River Delta. RESULTS: Two beta(s) genes, in apparently non-related individuals, were found. HPFH, detected in this same population of Warao Indians 25 years ago, was present in heterozygous form in five individuals from a large kindred, with hemoglobin F levels ranging from 3.7% to 8%, and with a pancellular distribution. The HPFH mutation was of the deletional type. beta-globin gene haplotypes were determined by direct counting (through family studies) in 150 chromosomes; 26% of the 150 examined cluster presented haplotype 2, 22% haplotype 6, and 13% a new, Warao haplotype. Haplotype 3, of probable African origin, was also found with a frequency of 5%. CONCLUSIONS: The presence of the HPFH mutation was confirmed, and the new beta-globin gene haplotype together with the presence of other rare variants indicates that the Warao are very distinctive in relation to other Native Americans. Evidence was also found of a slight admixture from Africa derived subjects (Layrisse et al. 1988). PMID- 18428015 TI - Obesity and regional fat distribution in Kenyan populations: impact of ethnicity and urbanization. AB - BACKGROUND: Obesity is increasing rapidly in Africa, and may not be associated with the same changes in body composition among different ethnic groups in Africa. OBJECTIVE: To assess abdominal visceral and subcutaneous fat thickness, prevalence of obesity, and differences in body composition in rural and urban Kenya. SUBJECTS AND METHODS: In a cross-sectional study carried out among Luo, Kamba and Maasai in rural and urban Kenya, abdominal visceral and subcutaneous fat thicknesses were measured by ultrasonography. Height and weight, waist, mid upper arm circumferences, and triceps skinfold thickness were measured. Body mass index (BMI), arm fat area (AFA) and arm muscle area (AMA) were calculated. RESULTS: Among 1430 individuals (58.3% females) aged 17-68 years, abdominal visceral and subcutaneous fat, BMI, AFA and waist circumference (WC) increased with age, and were highest in the Maasai and in the urban population. AMA was only higher with increasing age among males. The prevalence of overweight (BMI > or = 25) (39.8% vs. 15.8%) and obesity (BMI > or = 30) (15.5% vs. 5.1%) was highest in the urban vs. rural population. CONCLUSION: Abdominal visceral and subcutaneous fat thickness was higher with urban residency. A high prevalence of overweight and obesity was found. The Maasai had the highest overall fat accumulation. PMID- 18428018 TI - Drug-eluting stents - what should be improved? AB - Despite the success of drug-eluting stents (DES) in reducing restenosis and the need for target vessel revascularization, several deficiencies have been unraveled since their first clinical application including the risk of stent thrombosis, undesired effects due to the stent polymer as well as the stent itself, and incomplete inhibition of restenosis (especially in complex lesions). Several novel stent systems are being investigated in order to address these issues. In second-generation DES, the rapamycin analogues zotarolimus and everolimus (and more recently biolimus) have been most extensively studied. Furthermore, special stent-coatings to actively promote endothelial healing (in order to reduce the risk of stent thrombosis) and to further reduce restenosis have been employed. To avoid undesirable effects of currently applied (durable) polymers, biocompatible and bioabsorbable polymers as well as DES delivery systems without the need for a polymer have been developed. Bioabsorbable stents, both polymeric and metallic, were developed to decrease potential late complications after stent implantation. Although most of these innovative novel principles intuitively seem appealing and demonstrate good results in initial clinical evaluations, long-term large-scale studies are necessary in order to reliably assess whether these novel systems are truly superior to first generation DES with respect to safety and efficacy. PMID- 18428019 TI - The assembly of apoB-containing lipoproteins: a structural biology point of view. AB - Atherosclerosis is a widespread disease caused by the deposition of lipids on arterial walls. Such lipid plaques in coronary arteries can be fatal. Although many factors related to diet, life-style, etc. contribute to the worsening of the ailment, the primary cause, the lipids in the circulatory system, come from a series of low-density lipoproteins. These lipoproteins are necessary for the transport of lipids to and from different organs. It would be valuable to medicine and the field of drug design if a more detailed understanding of the organization of lipid and protein in these molecules were available. Unfortunately because of heterogeneity in their size and lipid composition, all classes of the low-density serum lipoproteins appear to be not amenable to the most widely used method for obtaining detailed atomic data - X-ray crystallography. However there appears to be a recently identified homolog that is relatively homogeneous, and crystal structures have been obtained. Used as a molecular model, the homolog serves as a source of conformational information that might help to unravel the processes involved in the lipid loading of the low density lipoproteins. The review attempts to give a brief summary of the structural biology of the serum low-density lipoproteins relative to the molecular model of lipovitellin. PMID- 18428020 TI - A review of rapid prototyping techniques for tissue engineering purposes. AB - Rapid prototyping (RP) is a common name for several techniques, which read in data from computer-aided design (CAD) drawings and manufacture automatically three-dimensional objects layer-by-layer according to the virtual design. The utilization of RP in tissue engineering enables the production of three dimensional scaffolds with complex geometries and very fine structures. Adding micro- and nanometer details into the scaffolds improves the mechanical properties of the scaffold and ensures better cell adhesion to the scaffold surface. Thus, tissue engineering constructs can be customized according to the data acquired from the medical scans to match the each patient's individual needs. In addition RP enables the control of the scaffold porosity making it possible to fabricate applications with desired structural integrity. Unfortunately, every RP process has its own unique disadvantages in building tissue engineering scaffolds. Hence, the future research should be focused on the development of RP machines designed specifically for fabrication of tissue engineering scaffolds, although RP methods already can serve as a link between tissue and engineering. PMID- 18428022 TI - Failure of aspirin to prevent myocardial infarction and adverse outcome during follow-up - a large series of all-comers. AB - BACKGROUND: Aspirin has been used for secondary prevention of myocardial infarction (MI) in individuals with coronary disease. Although supported by several large controlled trials, aspirin prevents only a portion of recurrent events. AIM AND METHOD: We set out to study the prevalence of long-term aspirin use prior to admission for MI and its significance for medium-term event-free survival; 998 consecutive patients with acute MI admitted to a tertiary center were included in the study. RESULTS: Nearly half (42.4%) of all patients reported long-term use of low-dose aspirin prior to the index event. Prior aspirin use was associated with a 50% increase in the risk of both the combined end point of recurrent unstable angina, recurrent myocardial infarction, stroke, or death (OR 1.49; 95% CI 1.12-2.00, P=0.006) and mortality (OR 1.50; 95% CI 1.03-2.17, P=0.03) during 10-month follow-up. Prior aspirin use was not associated with an increased frequency of added antithrombotic therapy at discharge. CONCLUSIONS: We have found that prior aspirin use is common in patients hospitalized for acute MI. Individuals already on aspirin had increased risk of recurrent ischemic events and all-cause mortality during the 10-month follow-up after their index MI. PMID- 18428023 TI - Long-term repeatability of measures of early insulin secretion derived from an intravenous glucose tolerance test and conversion from impaired glucose tolerance to diabetes. AB - AIM: We assessed the long-term repeatability of the acute insulin response (AIR) and sensitivity index (S(I)) derived from the frequently sampled intravenous glucose tolerance test (FSIGT). METHODS: An FSIGT was performed in 20 women who participated in a 6.5-month rye- and wheat-bread intervention trial, 70 men and women with impaired fasting glycaemia (IFG) or impaired glucose tolerance (IGT) who participated in the Genobin study, and 81 men and women with IGT who participated in the Finnish Diabetes Prevention Study (DPS). RESULTS: The correlation of AIR and S(I) at base-line with respective values after the 6.5-8.5 month trials was 0.86-0.88 and 0.71-0.84, and before and after 4 years in the DPS substudy, 0.86 and 0.53. In multivariate analyses, AIR (relative risk for a 1-SD change, 0.67; 95% confidence intervals 0.46-0.97) predicted the conversion from IGT to diabetes in the DPS substudy. CONCLUSION: AIR is highly repeatable even after 4 years of follow-up. The long-term repeatability of S(I) is moderate. Our findings emphasize the importance of impaired early insulin secretion in the transition from IGT to diabetes, and the high degree of tracking of measures of early insulin secretion derived from the FSIGT. PMID- 18428021 TI - Mitochondrial involvement in psychiatric disorders. AB - Recent findings of mitochondrial abnormalities in brains from subjects with neurological disorders have led to a renewed search for mitochondrial abnormalities in psychiatric disorders. A growing body of evidence suggests that there is mitochondrial dysfunction in schizophrenia, bipolar disorder, and major depressive disorder, including evidence from electron microscopy, imaging, gene expression, genotyping, and sequencing studies. Specific evidence of dysfunction such as increased common deletion and decreased gene expression in mitochondria in psychiatric illnesses suggests that direct examination of mitochondrial DNA from postmortem brain cells may provide further details of mitochondrial alterations in psychiatric disorders. PMID- 18428024 TI - Urinary matrix metalloproteinase -8, -9, -14 and their regulators (TRY-1, TRY-2, TATI) in patients with diabetic nephropathy. AB - Matrix metalloproteinase-9 (MMP-9) has been shown to be involved in the development of diabetic nephropathy (DNP). We studied the levels, molecular forms, and degree of activation of urinary MMP-8, -9, -14, trypsin-1 and -2, as well as tumor-associated trypsin inhibitor (TATI) of DNP patients and healthy controls. Urinary samples were analyzed for MMPs by Western blotting and gelatin zymography and for trypsin-1, -2, and TATI by time-resolved immunofluorometric assays. Total MMP-8 immunoreactivity, the proportion of active MMP-9, and gelatinolytic activity in urine were significantly higher in DNP patients than in controls. In urine of DNP patients the proportion of active polymorphonuclear neutrophil (PMN)-type (but not fibroblast-type) MMP-8 was increased. MMP-8 and MMP-9 were found to form high molecular weight complexes in DNP urine. Total immunoreactivity of soluble urinary MMP-14 and the levels of trypsin (TRY)-1 and TRY-2, but not of TATI, were also significantly increased in DNP. Zymography, Western blotting, and immunofluorometric analysis of DNP urine showed a significant association especially between activation of MMP-9 as well as PMN type MMP-8 and TRY-2. Our findings suggest that a trypsin-MMP cascade is involved in the pathogenesis of DNP, which may offer new possibilities for diagnosis and treatment of DNP with MMP inhibitors. PMID- 18428025 TI - Beneficial synergistic interactions of TNF-alpha and IL-6 in C2 skeletal myoblasts--potential cross-talk with IGF system. AB - The interaction effects of tumour necrosis factor-alpha (TNF-alpha) and interlukin-6 (IL-6) on skeletal muscle proliferation and differentiation remains controversial. We therefore investigated the potential interactive effects of TNF alpha and IL-6 on murine C2 skeletal myoblast survival, differentiation and proliferation. A novel and unexpected positive temporal interaction between TNF alpha and IL-6 on cell growth was identified (90%), with maximal beneficial effects obtained in myoblasts treated with TNF-alpha (10 ng/ml) for 24 h prior to being dosed with IL-6 (2.5 ng/ml) for a further 24 h. This combined treatment significantly (p < 0.05) increased the level of total cellular protein (330%), extracellular signal-regulated kinase (ERK) phosphorylation (55%), and S-phase of cell cycle (2.5-fold), confirming cell growth. The expression of mRNAs of key regulators of muscle mass: insulin-like growth factor binding protein-5, insulin like growth factor-II (IGF-II), IGF-I receptor (IGF-IR) and IGF-II receptor (IGF IIR) were also significantly (p < 0.05) increased by 1600-, 1.6-, 27- and 6-fold, respectively, giving an indication of the regulatory mechanisms of this interaction. Moreover, in response to this treatment, the expression level of signal-transducing glycoprotein 130 (gp130) was induced up to 3.5-fold but not after either treatments alone. This may not only explain the beneficial effects of this treatment on skeletal myoblast numbers but also define a functional role of gp130 in skeletal muscle cells. Our data suggest that in the presence of TNF alpha/IL-6 functions positively and potentially also cooperatively with the IGF system to achieve the maximal beneficial effect on skeletal myoblast numbers. PMID- 18428026 TI - The effects of inflammatory response associated with traumatic spinal cord injury in cutaneous wound healing and on expression of transforming growth factor-beta1 (TGF-beta1) and platelet-derived growth factor (PDGF)-A at the wound site in rats. AB - At the cellular level, spinal cord injury (SCI) provokes an inflammatory response that generates substantial secondary damage within the cord, but also may contribute to its repair. The aim of this study was to investigate the effects of inflammatory response associated with SCI in cutaneous wound healing and on expression of transforming growth factor-beta1 (TGF-beta1) and platelet-derived growth factor (PDGF)-A at the wound site in rats. At the 14th day analysis, the mean TGF-beta1 score in trauma group (I) was significantly lower than that in control group (C) (2.60 +/- 0.90 vs. 3.64 +/- 0.37, respectively; p < 0.05). The mean score for PDGF-A expression in group I was similar to the corresponding value in group C (2.42 +/- 0.74 vs. 2.94 +/- 0.72, respectively). Compared to group C, group I had significantly lower mean scores for epidermal and dermal regeneration, but higher mean scores for granulation tissue thickness and similar scores for angiogenesis. The dermal layer contains diffuse deposition of collagen fibers that are not organised as in control rat skin, and intraepidermal and subepidermal vasocongestion is distinct. Based on the results on the parameters evaluated in the study, experimental SCI in rats results in delay in wound healing and low intensity of TGF-beta1 in the dorsal wound-tissue specimens. PMID- 18428027 TI - Connective tissue growth factor: structure-function relationships of a mosaic, multifunctional protein. AB - Connective tissue growth factor (CTGF) is a member of the CCN family of six small secreted, cysteine-rich growth factors. The unique modular structure encompasses distinct functional domains which enable CTGF to interact with growth factors, surface receptors and matrix components. Widely expressed, CTGF has critical roles in embryonic development and the maintenance of normal cell and connective tissue function. It is also important for tissue repair following injury, and has been implicated in common diseases including atherosclerosis, pulmonary and renal fibrotic disorders and cancer. Factors that regulate CTGF transcription in response to exogenous stimuli, as well as downstream signalling pathways, have been described. However, only recently have studies begun to unravel how the functional domains within the CTGF modules orchestrate signals and control key biological processes. This article highlights how the structural and functional domains of CTGF and CTGF cleavage fragments integrate multiple extracellular events into cell signals. PMID- 18428029 TI - Regulation of DNA synthesis in mouse embryonic stem cells by transforming growth factor-alpha: involvement of the PI3-K/Akt and Notch/Wnt signaling pathways. AB - This study examined the mechanisms by which transforming growth factor (TGF) alpha regulates proliferation of mouse embryonic stem (ES) cells. TGF-alpha increased [3H] thymidine and BrdU incorporation in a time- (0-72 h) and dose dependent (0-10 ng/ml) manner. TGF-alpha stimulated the phosphorylation of Akt, mammalian target of rapamycin (mTOR), p70S6K1 and p44/42 mitogen-activated protein kinases (MAPKs). TGF-alpha also increased the protein levels of Notch, Notch intracellular domain, Hes-1 and Wnt1. However, TGF-alpha-induced DNA synthesis was blocked by inhibition of Akt, mTOR, p44/42 MAPKs and Notch. TGF alpha increased the gene expression of c-jun, c-myc and c-fos. Moreover, TGF alpha increased cyclin D/CDK 4 and cyclin E/CDK 2 levels, while decreasing p21cip1/waf1 and p27kip1, which were blocked by the inhibition of Akt, mTOR and Notch. In conclusion, TGF-alpha regulated DNA synthesis of mouse ES cells via PI3 K/Akt, p44/42 MAPKs and Notch/Wnt pathways. PMID- 18428028 TI - Precursor IGF-II (proIGF-II) and mature IGF-II (mIGF-II) induce Bcl-2 And Bcl-X L expression through different signaling pathways in breast cancer cells. AB - IGF-II plays a crucial role in fetal and cancer development by signaling through the IGF-I receptor. We have shown that inhibition of IGF-II by resveratrol (RSV) induced apoptosis and that proIGF-II (highly expressed in cancer) was more potent than mIGF-II in inhibiting this effect. Thus, we hypothesized that IGF-II differentially regulates the signaling cascade of the IGF-I receptor to stimulate the anti-apoptotic proteins Bcl-2 and Bcl-X(L) to prevent apoptosis. RSV treatment to breast cancer cells inhibited Bcl-2 and Bcl-X(L) expression and induced mitochondrial membrane depolarization. ProIGF-II was more potent than mIGF-II in: (1) activating the PI3/Akt pathway, (2) regulating Bcl-2 and Bcl-X(L) expression, and (3) inducing phosphorylation/nuclear translocation of Cyclic AMP responsive element binding protein. Furthermore, IGF-II differentially regulated the intracellular translocation of Bcl-2 and Bcl-X(L), a critical process in breast cancer progression to hormone-independence. Our study provides a novel mechanism of how proIGF-II promotes progression and chemoresistance in breast cancer development. PMID- 18428030 TI - Helmsman is expressed in both trachea and photoreceptor development: partial inactivation alters tracheal morphology and visually guided behavior. AB - We have identified helmsman (hlm), which is expressed in the fruit fly photoreceptor cells during neural network development. Hlm is also expressed in the elongating cells of the embryonic trachea. Both photoreceptor neurons and embryonic trachea cells elongate in precise, targeted growth for cell-to-cell specific recognition. Expression of antisense hlm-interfering RNA during embryogenesis arrests elongation of the developing tracheal cells and blocks maturation. Expression of hlm-interfering RNA during visual system formation results in reduced visual acuity and poor performance in optomotor response, indicative of abnormal neural network development. Hlm is a unique cell surface protein with complement-like protein interaction motifs. We have also cloned hlm from Lucilia cuprina (Australian blowfly), which is approximately 100 million years divergent from Drosophila, and find a remarkable 90% protein identity over the entire 558 amino acid protein. Analysis of the hlm sequence found in other species indicates a significant evolutionary pressure to maintain the hlm protein sequence. Our interpretation is that hlm is involved in cell maturation in both the elongating trachea and elongating photoreceptor cells. Cell adhesion and cell signaling, which are known to use immunoglobulin-like cell adhesion molecules, may use molecular systems analogous to complement to create protein complexes to regulate growth. PMID- 18428032 TI - SiO2 aerosol nanoparticle reactor for occupational health and safety studies. AB - Important questions are emerging about potential occupational safety, toxicological, and ecotoxicological effects and occupational inhalation exposure risks to engineered aerosol nanoparticles. Although multiple avenues are available to synthesize nanoparticles, few tools are accessible to industrial hygienists and inhalation toxicologists to produce well-characterized aerosols of known aerosol size distribution and particle number concentration that are stable, simple, and robust to operate. This article describes a SiO(2) aerosol nanoparticle reactor that has been developed as a tool for the study of the safety, health, and environmental consequences of exposure to nanoparticle synthesis and processing. The SiO(2) aerosol nanoparticle reactor is capable of stable, long-term synthesis of amorphous SiO(2) aerosol nanoparticles from d(50) = 10-70 nm at particle concentrations approximately 10(4)-10(7)particles/cm(3) that does not produce halogen-containing byproducts and does not require daily monitoring of the particle size distribution. This reactor is designed to produce a well-characterized aerosol to enable subsequent testing with a continuous, stable supply of aerosol nanoparticles (i) to facilitate inhalation toxicology studies, (ii) to measure explosion characteristics of aerosol nanoparticles, (iii) to determine the barrier efficacy for respirator filtration, bag house exhaust, and personal protective garment media challenged with diverse aerosol nanoparticles, and (iv) to develop airborne monitoring technologies for verifying workplace safety protocols. This article details reactor design, synthesis parameters, and instruments available to characterize the resulting aerosol nanoparticle size distributions. PMID- 18428031 TI - Effects of hyperkinetic, a beta subunit of Shaker voltage-dependent K+ channels, on the oxidation state of presynaptic nerve terminals. AB - The Drosophila Hyperkinetic (Hk) gene encodes a beta subunit of Shaker (Sh) K+ channels and shows high sequence homology to aldoketoreductase. Hk mutations are known to modify the voltage dependence and kinetics of Sh currents, which are also influenced by the oxidative state of the N-terminus region of the Sh channel, as demonstrated in heterologous expression experiments in frog oocytes. However, an in vivo role of Hk in cellular reduction/oxidation (redox) has not been demonstrated. By using a fluorescent indicator of reactive oxygen species (ROS), dihydrorhodamine-123 (DHR), we show that the presynaptic nerve terminal of larval motor axons is metabolically active, with more rapid accumulation of ROS in comparison with muscle cells. In Hk terminals, DHR fluorescence was greatly enhanced, indicating increased ROS levels. This observation implicates a role of the Hk beta subunit in redox regulation in presynaptic terminals. This phenomenon was paralleled by the expected effects of the mutations affecting glutathione S transferase S1 as well as applying H2O2 to wild-type synaptic terminals. Thus, our results also establish DHR as a useful tool for detecting ROS levels in the Drosophila neuromuscular junction. PMID- 18428033 TI - Low-level lead exposure among South Korean lead workers, and estimates of associated risk of cardiovascular diseases. AB - This study investigated the distribution of blood lead (PbB) levels, especially low levels, among Korean lead workers. The authors also estimated the potential effects of PbB on the blood pressure (BP) and cardiovascular diseases using models taken from the published meta-analyses. The PbB data from a total of 13,043 lead workers in 1217 companies throughout Korea were used. The geometric mean PbB level was 6.08 microg/dL (geometric standard deviation was 2.53), and 56.6% of the workers showed PbB levels greater than 5 microg/dL. Females accounted for 31.3% of all Korean lead workers in 2003. Considering two factors, such as PbB levels and the number of lead workers, the relatively important industry subclasses were identified as Manufacture of Accumulators; Manufacture of Other Electronic Valves, Tubes, Electronic Components n.e.c.; and Manufacture of Other Parts and Accessories for Motor Vehicles. The industrial processes of relative importance included battery assembly, acid treatment, and other soldering. Although uncertainties exist in the prediction model and associated model parameters, the authors attempted to estimate potential adverse health effects related to the lead exposure. It was estimated that 7383 South Korean lead workers might have increased blood pressure and the health risks due to the lead exposures in 2003. The highest estimated risk of BP increases due to lead exposures was expected in workers of industrial subclasses and processes, such as Other Basic Nonferrous Metal Industries and Maintenance. If the models in this study were applied to the South Korean population, the impact fractions for cardiac disease among lead workers would be estimated at 4.9-12.8 times those of the general population. PMID- 18428034 TI - Advances in functional regulation mechanisms of plant aquaporins: their diversity, gene expression, localization, structure and roles in plant soil-water relations (Review). AB - Aquaporins are important molecules that control the moisture level of cells and water flow in plants. Plant aquaporins are present in various tissues, and play roles in water transport, cell differentiation and cell enlargement involved in plant growth and water relations. The insights into aquaporins' diversity, structure, expression, post-translational modification, permeability properties, subcellular location, etc., from considerable studies, can lead to an understanding of basic features of the water transport mechanism and increased illumination into plant water relations. Recent important advances in determining the structure and activity of different aquaporins give further details on the mechanism of functional regulation. Therefore, the current paper mainly focuses on aquaporin structure-function relationships, in order to understand the function and regulation of aquaporins at the cellular level and in the whole plant subjected to various environmental conditions. As a result, the straightforward view is that most aquaporins in plants are to regulate water flow mainly at cellular scale, which is the most widespread general interpretation of the physiological and functional assays in plants. PMID- 18428035 TI - Influence of the passenger domain of a model autotransporter on the properties of its translocator domain. AB - Autotransporters are a superfamily of proteins secreted by Gram-negative bacteria including many virulence factors. They are modular proteins composed of an N terminal signal peptide, a surface-exposed 'passenger' domain carrying the activity of the protein, and a C-terminal 'translocator' domain composed of an alpha-helical linker region and a transmembrane beta-barrel. The translocator domain plays an essential role for the secretion of the passenger domain across the outer membrane; however, the mechanism of autotransport remains poorly understood. The whooping cough agent Bordetella pertussis produces an autotransporter serine-protease, SphB1, which is involved in the maturation of an adhesin at the bacterial surface. SphB1 also mediates the proteolytic maturation of its own precursor. We used SphB1 as a model autotransporter and performed the first comparisons of the biochemical and biophysical properties of an isolated translocator domain with those of the same domain preceded by the C-terminal moiety of its natural passenger. By using cross-linking and dynamic light scattering, we provide evidence that the passenger domain promotes the auto association of SphB1, although these interactions appear rather labile. Electrophysiological studies revealed that the passenger domain of the autotransporter appears to maintain the translocator channel in a low-conductance conformation, most likely by stabilizing the alpha-helix inside the pore. That the passenger may significantly influence AT physicochemical properties is likely to be relevant for the in vivo maturation and stability of AT proteins. PMID- 18428036 TI - EAAT2 density at the astrocyte plasma membrane and Ca(2 + )-regulated exocytosis. AB - We studied whether regulated exocytosis affects the glutamate transporter density in cultured astrocytes, in which the expression of a fluorescently labeled excitatory amino acid transporter 2 (EAAT2-EGFP) predominantly labeled the plasma membrane. The addition of ionomycin that elevates cytosolic Ca(2+) strongly increased the fluorescence of FM 4-64 membrane area dye, confirming the presence of regulated exocytosis in transfected astrocytes. However, concomitant with Ca(2+)-dependent FM 4-64 fluorescence increase, ionomycin induced a significant steady-state decrease in EAAT2-EGFP fluorescence. This is likely due to a secondary inner filter effect since,(i) in the absence of FM 4-64, ionomycin stimulation was ineffective in changing the EAAT2-EGFP fluorescence, and (ii) fluorescence changes in FM 4-64 and EAAT2-EGFP were inversely correlated. To test whether subcellular EAAT2-EGFP structures are translocated from the cytoplasm to the plasma membrane during ionomycin stimulation, EAAT2-EGFP fluorescence was monitored locally at the plasma membrane and a few microns away in the adjacent cytoplasm. Measurements revealed sites with an increase in EAAT2-EGFP plasma membrane fluorescence correlated with a fluorescence decrease beneath the plasma membrane, and sites with plasma membrane fluorescence decrease correlated with fluorescence increase within the adjacent cytoplasm. The sites of rapid translocation/retrieval of EAAT2-EGFP structures to/from the plasma membrane appeared to be distributed in a punctuate pattern around the cell perimeter. The density of EAAT2-EGFP was regulated in a Ca(2+)-dependent manner, since in the absence of extracellular Ca(2+) local translocation/retrieval events were absent, revealing rapid surface density regulation of EAAT2 in astrocytes by regulated exo/endocytosis. PMID- 18428037 TI - Effects of cholesterol levels on the excitability of rat hippocampal neurons. AB - Changes in the cholesterol levels dynamically alter the microenvironment of the plasma membrane and have been shown to modify functions of ion channels. However, the cellular effect of these modifications is largely unknown. In this report, we demonstrate that cholesterol levels modulate neuronal excitability in rat hippocampal neurons. Reduction of cholesterol levels shortened the duration and increased the firing frequency and peak amplitude of action potentials, while enrichment of cholesterol reversed the effect. Furthermore, we showed that reduction of cholesterol levels increased, while enrichment of cholesterol decreased the amplitude of the delayed rectifier I(K) currents. On the other hand, reduction of cholesterol levels slowed down the inactivation of the fast transient I(A) currents, but enrichment of cholesterol had no significant effect on the I(A) currents. Besides, alteration in cholesterol levels had no significant effect on the action potential in the presence of blockers for both I(K) and I(A) currents. These observations demonstrate that cholesterol levels bi directionally regulate the neuronal excitability mainly through modifications of the I(K) and I(A) currents, suggesting an optimum level of cholesterol for the optimum excitability of neurons. Alterations in the neuronal cholesterol levels have been associated with aging, cognitive decline, neurodegenerative diseases, etc. Therefore, our findings are important for a deeper understanding of the relationship between the cholesterol level and dysfunctions of the brain at the molecular level. PMID- 18428039 TI - Structural-dynamical properties of the transmembrane segment VI of the mitochondrial oxoglutarate carrier studied by site directed spin-labeling. AB - Site directed spin-labeling (SDSL) has been used to probe the structural and dynamic features of residues comprising the sixth transmembrane segment of the mitochondrial oxoglutarate carrier. Starting from a functional carrier, where cysteines have been replaced by serines, 18 consecutive residues (from G281 to I298) have been mutated to cysteine and subsequently labeled with a thiol selective nitroxide probe. The labeled proteins, reconstituted into liposomes, have been assayed for their transport activity and analyzed with continuous-wave electron paramagnetic resonance. Linewidth analysis, that is correlated to local probe mobility, indicates a well defined periodicity of the whole segment from G281 to I298, indicating that it has an alpha-helical structure. Saturation behaviour, in presence of paramagnetic perturbants of different hydrophobicities, allow the definition of the polarity of the individual residues and to assign their orientation with respect to the lipid bilayer or to the water accessible translocation channel. Comparison of the EPR data, homology model and activity data indicate that the segment is made by an alpha helix, accommodated in an amphipathic environment, partially distorted in the middle at the level of L289, probably because of the presence of a proline residue (P291). The C-terminal region of the segment is less restrained and more flexible than the N-terminus. PMID- 18428038 TI - Targeted disruption of Slc2a8 (GLUT8) reduces motility and mitochondrial potential of spermatozoa. AB - GLUT8 is a class 3 sugar transport facilitator which is predominantly expressed in testis and also detected in brain, heart, skeletal muscle, adipose tissue, adrenal gland, and liver. Since its physiological function in these tissues is unknown, we generated a Slc2a8 null mouse and characterized its phenotype. Slc2a8 knockout mice appeared healthy and exhibited normal growth, body weight development and glycemic control, indicating that GLUT8 does not play a significant role for maintenance of whole body glucose homeostasis. However, analysis of the offspring distribution of heterozygous mating indicated a lower number of Slc2a8 knockout offspring (30.5:47.3:22.1%, Slc2a8(+/+), Slc2a8(+/-), and Slc2a8(-/-) mice, respectively) resulting in a deviation (p=0.0024) from the expected Mendelian distribution. This difference was associated with lower ATP levels, a reduced mitochondrial membrane potential and a significant reduction of sperm motility of the Slc2a8 knockout in comparison to wild-type spermatozoa. In contrast, number and survival rate of spermatozoa were not altered. These data indicate that GLUT8 plays an important role in the energy metabolism of sperm cells. PMID- 18428040 TI - Membrane adsorption, folding, insertion and translocation of synthetic trans membrane peptides. AB - Spontaneous membrane adsorption, folding and insertion of the synthetic WALP16 and KALP16 peptides was studied by computer simulations starting from completely extended conformations. The peptides were simulated using an unmodified all-atom force field in combination with an efficient Monte Carlo sampling algorithm. The membrane is represented implicitly as a hydrophobic zone inside a continuum solvent modelled using the generalized Born theory of solvation. The method was previously parameterized to match insertion energies of hydrophobic side chain analogs into cyclohexane and no parameters were optimized for the present simulations. Both peptides rapidly precipitate out of bulk solution and adsorb to the membrane surface. Interfacial folding into a helical conformation is followed by membrane insertion. Both the peptide conformations and their location in the membrane are strongly temperature dependent. The temperature dependent behaviour can be summarized by fitting to a four-state model, separating the system into folded and unfolded conformers, which are either inserted into the membrane or located at the interfaces. As the temperature is lowered the dominant peptide conformation of the system changes from unfolded surface bound configurations to folded surface bound states. Folded trans-membrane conformers represent the dominant configuration at low temperatures. The analysis allows direct estimates of the free energies of peptide folding and membrane insertion. In the case of WALP the quality of the fit is excellent and the thermodynamic behaviour is in good agreement with expected theoretical consideration. For KALP the fit is more problematic due to the large solvation energies of the charged lysine residues. PMID- 18428041 TI - Patching of ganglioside(M1) in human erythrocytes - distribution of CD47 and CD59 in patched and curved membrane. AB - Membrane rafts may act as platforms for membrane protein signalling. Rafts have also been implicated in the sorting of membrane components during membrane budding. We have studied by fluorescence microscopy cross-linking of ganglioside GM1 in the human erythrocyte membrane, and how membrane proteins CD47 and CD59 distribute in GM1 patched discoid cells and calcium-induced echinocytic cells. Patching of ganglioside(M1) (GM1) by cholera toxin subunit B (CTB) plus anti-CTB resulted in the formation of usually 40-60 GM1 patches distributed over the membrane in discoid erythrocytes. Pre-treatment of erythrocytes with methyl-beta cyclodextrin abolished GM1 patching. GM1 patching was insensitive to pre-fixation (paraformaldehyde) of cells. Patching of GM1 did not affect the discoid shape of erythrocytes. Membrane proteins CD47 and CD59 did not accumulate into GM1 patches. No capping of patches occurred. GM1 accumulated in calcium-induced echinocytic spiculae. Also CD59, but not CD47, accumulated in spiculae. However, CD59 showed a low degree of co-localization with GM1 and frequently accumulated in different spiculae than GM1. In conclusion, our study describes a novel method for examining properties and composition of rafts. The study characterizes raft patching in the human erythrocyte membrane and emphasizes the mobility and 'echinophilicity' of GM1. Glycosyl phosphatidylinositol-anchored CD59 was identified as a mobile 'echinophilic' but 'raftophobic(GM1)' protein. Largely immobile CD47 showed no segregation. PMID- 18428042 TI - Tell haematologists there's been no progress in CML and ... see 'em yell! PMID- 18428043 TI - Current and emerging tests for the laboratory monitoring of chronic myeloid leukaemia and related disorders. AB - Chronic myeloid leukaemia (CML) is a molecularly defined disease. The BCR-ABL fusion occurs in all cases of classical CML and leukaemic cells express a constitutively activated BCR-ABL tyrosine kinase. Other fusion oncogenes involving tyrosine kinases, including ABL and PDGFRA/B, have been identified, and are associated with leukaemic syndromes that may resemble CML. The discovery and treatment of these related disorders has been facilitated by our detailed understanding of CML. Imatinib mesylate has significantly improved the outcome of patients with CML, but there remains a significant minority of chronic phase CML patients for whom the response to treatment with standard dose imatinib is suboptimal. Cytogenetic and molecular monitoring of the response to treatment provides important prognostic information. Achievement of a major molecular response (MMR) in chronic phase patients treated de novo with imatinib confers near 100% freedom from progression to advanced phase, and MMR is now an important goal of therapy. Standardisation of BCR-ABL molecular monitoring is under way and should enable the accurate and reproducible identification of MMR in laboratories around the world. Point mutations in the kinase domain of BCR-ABL are the most common cause of acquired resistance to imatinib treatment. The susceptibility of a mutation to imatinib, nilotinib, or dasatinib may help to guide changes in therapy in a patient with resistance. In addition to these established methods of monitoring, there are new tests in development that may assist in determining prognosis and optimising therapy. Among patients receiving the same dose of imatinib, the plasma level of imatinib shows considerable inter-patient variation, and there is emerging evidence that higher levels may be associated with improved response to treatment. The intracellular concentration of imatinib also shows considerable variation, most likely related to differences in influx and efflux transport mechanisms. We discuss how these established and emerging assays might be used to optimise the treatment of CML patients. PMID- 18428044 TI - What's in a cancer syndrome? Genes, phenotype and pathology. AB - Syndromes are characterised by a group of specific signs and symptoms. This review aims to provide an overview of cancer syndromes and sheds light on possible roles for general health professionals in relation to their abilities to identify patients with cancer syndromes and therefore to refer such patients to specialists. PMID- 18428046 TI - Pulmonary pathology in patients associated with scrub typhus. AB - AIMS: Scrub typhus is a zoonotic disease caused by Orientia tsutsugamushi. Severe cases resulting in mortality from this disease have rarely been reported. We present two scrub typhus cases (a man and a girl) who died of acute respiratory distress syndrome (ARDS). METHODS: Autopsies were performed. Histopathological and immunohistochemical stains were employed using specific antibody for O. tsutsugamushi and inducible nitric oxide synthase (iNOS). RESULTS: These subjects developed respiratory distress shortly after admission, and expired following respiratory failure. At autopsy, generalised lymphadenopathy was observed. The lung weight was about two-fold the normal value. Gross inspection revealed oedematous and haemorrhagic lungs. Microscopic examination revealed diffuse alveolar damage with hyaline membrane formation and interstitial pneumonitis with infiltration of inflammatory cells. Immunohistochemical stain showed O. tsutsugamushi antigen depositions in the endothelial cells. We also demonstrated iNOS in the alveolar macrophages and lung tissue debris in both cases. CONCLUSION: Scrub typhus is usually a mild infectious disease. Our cases present the most dramatic example of sudden death due to ARDS in a short period of time. The clinical investigation and analysis suggest direct endothelial cell invasion of the organism and marked iNOS expression may be involved in the pathogenesis of ARDS associated with scrub typhus. PMID- 18428045 TI - Bile duct to portal space ratio and ductal plate remnants in liver disease of infants aged less than 1 year. AB - AIM: To validate the bile duct to portal space ratio as an independent factor useful for the prognosis of neonatal liver disease. METHODS: We assessed the maturation of the intrahepatic bile duct system (IBDS) in 87 consecutive infants aged less than 1 year undergoing non-subcapsular, adequate (at least six portal tracts), liver needle biopsies because of hepatomegaly and/or cholestasis. The maturation of the IBDS was evaluated by immunohistochemistry with an antibody directed to cytokeratin 7 (CK7), a biliary-type intermediate filament of the cytoskeleton, and a schema showing the IBDS remodelling. We used five categories to fit the different patterns of the IBDS remodelling using the ratio between the number of bile ducts and the number of portal tracts (BD/PT) and the presence of abnormal reaction patterns (marked intra-acinar pseudorosettes and/or periportal ductular proliferation): (A) abnormal reaction patterns with any BD/PT; (B) BD/PT = 0; (C) 0.1 < or = BD/PT < 0.5; (D) 0.5 < or = BD/PT < 0.9; and (E) BD/PT > 0.9 (B-E categories: no abnormal reaction patterns). Further, we evaluated cholestasis, portal fibrosis (PF), portal inflammation (PI), giant cell transformation (GCT), and extramedullary haematopoiesis (EMH). RESULTS: We identified A-E categories in 24, 14, 17, 8, and 24 biopsies, respectively. B and C categories were frequently observed in biliary atresia (BA), A category in neonatal hepatitis (NH), A-C categories in paucity of intrahepatic bile ducts (PIBD), and E category in 'other liver diseases' (OLD). Cholestasis, PI, GCT, and EMH were more frequent in A and C, while PF was variably seen in all categories. The lowest survival rate occurred in B (Kaplan-Meier estimator). CONCLUSIONS: (1) Biliary epithelial cell patterns recapitulate the primitive stages of the IBDS maturation; (2) abnormal reaction patterns occur mainly in NH, whilst BD/PT < 0.5 in BA; and (3) lack of intrahepatic bile ducts in infants aged less than 1 year is an adverse prognostic factor independent from aetiology of neonatal liver disease. PMID- 18428047 TI - Transdifferentiation into biliary ductular cells of hepatocytes transplanted into the spleen. AB - AIMS: Transplantation of rat hepatocytes into the syngeneic rat spleen results in the appearance of cytokeratin (CK)7 and CK19 positive biliary cells that form ductules. We examined whether hepatocytes are the origin of these biliary ductular cells. METHODS: We transplanted rat dipeptidyl peptidase IV (DPPIV) positive hepatocytes into the liver of retrorsine-treated and partially hepatectomised DPPIV negative rats, which resulted in proliferation of DPPIV positive hepatocytes in the liver. Two months later, hepatocytes were prepared from chimaeric livers of these rats and transplanted into the spleen of DPPIV negative rats. Four weeks later, the expression of DPPIV in CK7 positive ductules in the spleen was examined by immunofluorescent double-staining. RESULTS: In the spleen of DPPIV negative rats transplanted with hepatocytes prepared from the chimaeric livers, DPPIV was found to be expressed in some CK7 positive biliary ductules where only a fraction of cells expressed DPPIV, whereas in the spleen of DPPIV negative rats transplanted with hepatocytes from livers of DPPIV positive rats, DPPIV was expressed in all CK7 positive biliary ductules. CONCLUSION: The present study indicates that hepatocytes transplanted into the spleen could transdifferentiate into biliary cells that aggregate to form ductular structures. PMID- 18428048 TI - Time to think outside the box? Prothrombin time, international normalised ratio, international sensitivity index, mean normal prothrombin time and measurement of uncertainty: a novel approach to standardisation. AB - BACKGROUND: The prothrombin time (PT) assay is the most clinically ordered coagulation test, and most often used for monitoring of vitamin K antagonist therapy (e.g., warfarin), where results are expressed as an international normalised ratio (INR). The INR is in essence the patient's PT 'mathematically adjusted' to a standardised value taking into account the peculiarities of the test system as defined by an ISI (international sensitivity index) and MNPT (mean normal prothrombin time). Although some manufacturers provide assigned ISI values for specific PT reagents and instrumentation, it is still recommended practice that laboratories check or validate these ISIs, as well as estimate the MNPT. Where an ISI is not provided by a manufacturer, the laboratory needs to estimate its own value. Current recommendations suggest the use of commercial reference plasma calibration sets, but there is limited information on the performance of these in the field. RESULTS: We report a comparative study that assessed the utility of three such commercial calibration plasma sets, used as recommended, as well as alternate or supplementary procedures for estimation of ISI and MNPT. The latter included one novel approach using comparative data of 'existing' versus 'replacement' reagent, as well as assessment of external quality assurance data. Although MNPT value estimates were not grossly disparate, a wide variety of ISI values (e.g., 1.12-1.30 for our primary instrument) was obtained with the different plasma sets. CONCLUSION: Because of the above, further verification checks are required prior to acceptance of ISI and MNPT estimates generated from commercial plasma calibration sets. We also provide some recommendations regarding the process of standardisation of INR testing. PMID- 18428049 TI - B-cell maturation defects in common variable immunodeficiency and association with clinical features. AB - AIMS: Patients with common variable immunodeficiency (CVID) often have defects in post-antigenic B-cell differentiation with fewer memory B cells and impaired isotype switching. We aimed to classify CVID patients according to these defects and determine whether this predicted clinical manifestations. METHODS: We analysed the memory marker CD27, maturation marker CD21, and IgD on peripheral blood B cells from 31 CVID patients and 23 controls using a whole-blood lysis technique, allocated patients according to two classifications ('Freiburg' and 'Paris') and correlated results with clinical manifestations. RESULTS: CVID patients had fewer memory (CD27(+)) B cells and isotype-switched (IgD(-)) memory B cells in absolute number and proportion. Many CVID patients had increased immature (CD21(-)) B cells. Lymphoproliferation and autoimmune cytopenias were found almost exclusively in these patients, including Freiburg group Ia (decreased switched memory and increased immature B cells), but also those with normal switched memory and increased immature B cells. The Paris classification was less useful in predicting clinical manifestations. CONCLUSIONS: CVID is associated with defects in memory B-cell differentiation. Subclassification helps identify patients with clinical manifestations, particularly lymphoproliferation and autoimmune cytopenias in those with impaired B-cell maturation and isotype switching. Routine B-cell phenotyping may assist clinicians in predicting these clinical features. PMID- 18428050 TI - Detection of BRAF V600E mutation by pyrosequencing. AB - INTRODUCTION: Detection of the V600E hotspot mutation in BRAF oncogene is extremely useful for the screening of hereditary non-polyposis colorectal cancer (Lynch's syndrome) and for the prediction of sensitivity to MEK inhibitors. Here we describe a method for detecting this mutation based upon pyrosequencing technology. METHODS: The efficiency of pyrosequencing for detecting BRAF V600E mutations was compared with the conventional dideoxy sequencing method in 12 tumour cell lines and in 108 colorectal tumours. RESULTS: The results from pyrosequencing were 100% concordant with those from dideoxy sequencing. This method was capable of detecting BRAF V600E mutations at a much lower ratio of mutant to wild-type alleles (1:50) than dideoxy sequencing (1:5) while being considerably faster and less expensive. CONCLUSIONS: Pyrosequencing offers a specific, sensitive, rapid and cost-effective alternative to dideoxy sequencing for the detection of BRAF V600E mutations in clinical tumour specimens. PMID- 18428051 TI - Test and teach. An uncommon but distinctive breast lesion. Diagnosis: Cystic hypersecretory lesion associated with pregnancy-like hyperplasia. PMID- 18428052 TI - Late gastric ulceration and cytomegalovirus infection following selective internal radiation therapy (SIRT) of the liver. PMID- 18428053 TI - hCDC4 gene mutation is rare in colorectal carcinomas in Korean patients. PMID- 18428054 TI - Renal cell carcinoma in a 33-year-old male with an unusual morphology and an aggressive clinical course: possible Xp11.2 translocation. PMID- 18428055 TI - An unusual glomerulopathy in a man with HIV infection on HAART. PMID- 18428056 TI - Lymphoepithelioma-like carcinoma of the urinary bladder. PMID- 18428057 TI - Soft tissue recurrence of sacral giant cell tumour of bone as an intra-abdominal mass: an unusual presentation. PMID- 18428058 TI - Giant chordoma occupying the whole abdominal cavity. PMID- 18428059 TI - Merkel cell carcinoma with fibrosarcomatous differentiation. PMID- 18428060 TI - Recurrent squamous cell carcinoma and follicular lymphoma arising in the scalp after treatment for lymphoma. PMID- 18428061 TI - Follicular Hodgkin's lymphoma recurring as nodular sclerosis Hodgkin's lymphoma: report of two cases. PMID- 18428062 TI - 'Breast-like' lesions in the vulva: histology, origin and significance. PMID- 18428063 TI - Proximal-type epithelioid sarcoma of the scrotum: a difficult diagnosis in an unusual location and review of the literature. PMID- 18428064 TI - Cellular angiofibroma of the scrotum. PMID- 18428065 TI - Tubulopapillary clear cell carcinoma of the stomach may be a type of pylorocardiac carcinoma. PMID- 18428067 TI - Inhibitory control deficits in childhood and risk for substance use disorders: a review. AB - Identification of neurobiological factors that confer risk for the development of addiction may substantially advance development of new prevention and treatment strategies to combat substance use disorders. This review focuses on the relationship between impulsivity - a behavior that is common to the clinical picture of both substance use disorders (SUD) and childhood disruptive behavior disorders - and neurobiological risk for SUD. It further examines various behaviors within the over-arching domain of impulsivity, ultimately focusing on the more narrowly defined and measurable construct of inhibitory control, and concluding that underlying deficits in inhibitory control may be central to many of the behaviors associated with high risk for SUD. Targeted cross-sectional study of the neural basis of inhibitory dyscontrol in subjects at high risk for SUD, who have not yet begun to abuse drugs, has the potential to generate important hypotheses regarding the neurobiological underpinnings of SUD risk. Hypotheses developed using this approach can be more definitively evaluated in longitudinal studies with these same populations, extending through the period of maximal risk for SUD in adolescence and early adulthood. PMID- 18428068 TI - Prospective relations between melancholia and substance use disorders. AB - Examining associations between subtypes of major depressive disorder (MDD) and substance use disorders (SUDs) might elucidate mechanisms of comorbidity between MDD and SUDs. This study evaluated prospective relations between SUDs and melancholic MDD. A cohort of community-dwelling participants with lifetime history of MDD (N = 460) were assessed for DSM-IV mental disorders using structured clinical interviews at ages 24 and 30. Stimulant use disorders and melancholic MDD were prospective risk factors for each other over the 6-year period following the age-24 assessment. Associations were robust when controlling for clinical severity/chronicity. Alcohol and cannabis use disorders were not robustly associated with melancholia. PMID- 18428069 TI - Tai-Kang-Ning, a Chinese herbal medicine formula, alleviates acute heroin withdrawal. AB - The efficacy and safety of Tai-Kang-Ning (TKN) capsule, a traditional Chinese medicine formula, for the treatment of acute heroin withdrawal syndrome were investigated by conducting a double-blind, double-dummy, positive-controlled, and randomized trial. Sixty-four patients with acute heroin withdrawal syndrome were recruited. These patients were treated with either TKN or lofexidine in a fixed schedule of doses for 10 days. The results indicate that both treatments significantly reduced withdrawal symptoms by day 3, but there was no significant difference overall between lofexidine and TKN in efficacy or safety. These results demonstrate that TKN is effective in the treatment of moderate-to-severe acute heroin withdrawal syndrome with mild adverse effects. PMID- 18428070 TI - Child sexual abuse, dissociation, and alcohol: implications of chemical dissociation via blackouts among college women. AB - The present study examined whether childhood sexual trauma moderated the relationships between dissociation and both problematic college drinking and alcohol-induced blackouts among a sample of college females (N = 156). Cross sectional data were consistent with the moderation hypotheses. Simple effects showed that the relationship between dissociation and blackout frequency as well as problematic drinking only existed among those with sexual trauma histories (p < .035), but not among those reporting no sexual trauma exposure (p = .333). Findings implicate the use of alcohol as a possible maladaptive coping mechanism among college females with childhood sexual trauma. PMID- 18428071 TI - Further validity evidence for the dimensions of change in therapeutic community treatment instrument. AB - Recent research efforts have focused on assessing the therapeutic community (TC) treatment process. The current article examines the validity of the client self report Dimensions of Change Instrument (DCI) treatment process measure, by confirming its factor structure and examining each subscale's relationship with time in treatment, scales from an existing process assessment, and mental and physical health status. Participants were 511 adult and adolescent residential TC clients. The factor structure of the DCI replicated with satisfactory fit. The pattern of associations between DCI subscales and criterion measures was largely as expected, providing initial concurrent and discriminant validity evidence for the DCI. PMID- 18428072 TI - Effects of potential agonist-replacement therapies for stimulant dependence on inhibitory control in cocaine abusers. AB - Two experiments were conducted to determine whether methylphenidate or modafinil, two potential pharmacotherapies for stimulant dependence, would impair inhibitory behavior in cocaine users. Eleven cocaine abusers were administered methylphenidate (0, 15, 30, and 45 mg) or modafinil (0, 150, 300, and 450 mg) across four experimental sessions. A cued go-no-go task was used to measure response execution and inhibition. Subjective and cardiovascular measures were collected. Neither methylphenidate nor modafinil impaired inhibitory control, but produced prototypical subject-rated and cardiovascular effects. The results of these studies may have implications for the use of these drugs as agonist replacement therapies for stimulant dependence. PMID- 18428073 TI - Peer environment mediates parental history and individual risk in the etiology of cannabis use disorder in boys: a 10-year prospective study. AB - Previous research has shown that a trait termed neurobehavior disinhibition (ND) measured in childhood predicts substance use disorder by young adulthood. The present investigation extends these findings by determining the degree to which peer environment mediates the association between ND and development of cannabis use disorder (CUD). ND was measured in a sample of 216 boys 10-12 years of age. The peer environment was assessed at age 16. Current CUD was determined at age 22. Paternal and maternal SUD predicted son's ND which, in turn, predicted son's peer environment and, subsequently, son's cannabis use frequency and CUD. Peer environment mediated the association between ND and cannabis use and ND and CUD. Maternal and paternal SUD predicted the peer environment. Parental SUD, son's ND, and son's peer environment predicted CUD at age 22 with 84% accuracy. PMID- 18428074 TI - Serum cortisol secretion during heroin abstinence is elevated only nocturnally. AB - Several studies indicate abnormalities in the hypothalamic-pituitary-adrenal axis (HPA) during acute opiate withdrawal, but protracted withdrawal has gotten less study. The current study further characterized the 24-hour time course of plasma cortisol levels in heroin-dependent individuals who were abstinent for 10-15 days, which is beyond the 5 days of acute withdrawal, compared to demographically matched healthy controls using samples collected every 3 hours over 24 hours and assessed with radioimmunoassay (RIA). The abstinent heroin-dependent participants had significantly higher plasma cortisol levels nocturnally suggesting a loss of diurnal variation in these heroin subjects. PMID- 18428075 TI - HIV risk and history of STDs in MCMI-III psychopathology subgroups of comorbid substance abusers. AB - The purpose of this investigation is to investigate HIV risk-related attitudes, beliefs, expectancies, behaviors, and histories of lifetime sexually transmitted diseases in the Millon Clinical Multiaxial Inventory III (MCMI-III) defined psychopathology cluster subgroups. Hierarchical agglomerative cluster analysis, using Ward's method, was employed that led to identification of high (n = 37), medium (n = 132), and low (n = 28) MCMI-III psychopathology cluster subgroups. Members of the low psychopathology subgroup demonstrated significantly higher levels of knowledge about HIV and AIDS and less anxiety about HIV infection than high and moderate psychopathology subgroups. The high psychopathology subgroup reported greater importance of approval of condom use by partners but less sexual self-efficacy than the moderate psychopathology subgroup. This high pathology group revealed less favorable condom attitudes than did the low psychopathology subgroup and a significantly higher percentage of unprotected vaginal sex acts in the past 6 months than did members of the other two subgroups. A comparatively low rate of lifetime syphilis was reported in the low psychopathology subgroup (all ps < .05). Results are discussed in the context of a growing literature indicating distinctive treatment needs among members of high psychopathology subgroups of drug treatment participants. PMID- 18428076 TI - Aggression, impulsivity, personality traits, and childhood trauma of prisoners with substance abuse and addiction. AB - INTRODUCTION: The aim of our study is then to analyze psychological and judicial features of a subgroup of inmates with substance abuse. METHODS: Prisoners with substance abuse (n = 312) were compared to prisoners without substance abuse (n = 591). Recruited inmates completed a semistructured interview for collection of sociodemographic and judicial data and a battery of psychometric tests for assessement of aggression, impulsivity, depression, personality traits, hostility, resilience, and childhood trauma. RESULTS: Substance abusers had on average multiple incarcerations (78.8%), more juvenile convictions (60.2%), more violent behaviors during detention (29.8%), and a history of one or more suicide attempts (20.8%). They also had higher scores on subscales for childhood trauma, higher scores for psychoticism and neuroticism, higher impulsivity levels, worse resilience, increased hostility, and prevalent suicidal ideation. CONCLUSION: Prisoners with substance abuse constitute a subgroup with increased judiciary and psychiatric issues, possibly due to early life history and psychological characteristics, such as high impulsivity and aggressiveness, poor resilience, and higher suicidal risk. PMID- 18428077 TI - Methadone- and heroin-related deaths in Florida. AB - Methadone is a potent synthetic opioid used for treatment of opioid dependence and chronic pain. Florida Department of Law Enforcement data were analyzed to examine trends in deaths related to or caused by methadone and/or heroin between 2001-2006. Results demonstrated that mortalities associated with methadone use increased steadily as mortalities associated with heroin decreased steadily. Though useful in the treatment of opioid dependence and chronic pain, methadone possesses high abuse potential and documented mortality risks. Treatment with methadone, for both pain and opioid dependence, should be preceded by an abuse liability evaluation. Attempts to minimize diversion should be implemented. PMID- 18428079 TI - Outcomes in major depressive disorder: the evolving concept of remission and its implications for treatment. AB - It is increasingly recognised that major depressive disorder can be a chronic condition with many patients experiencing recurrent episodes. Remission from a depressive episode implies the absence or near absence of depressive symptoms. However, for many patients the periods between depressive episodes are not symptom free. Residual symptoms are predictors of relapse or recurrence, and may be associated with residual psychosocial impairment. In clinical studies, remission is commonly defined using a cut-off score on a rating scale for depressive symptoms, such as a score of < or = 7 on the 17-item Hamilton scale. However, there is debate about which scales and cut-offs are optimal and full length rating scales are not widely used in clinical practice. In spite of such issues, it seems clear that a therapy should aim at the most complete remission possible. Unfortunately, recent studies have highlighted that in clinical practice usually only a low rate of remission is achieved. Although long-term treatment with antidepressants can reduce the risk of relapse or recurrence only a minority of patients in clinical practice achieve this as treatment is often prematurely stopped due to long-term side effects such as sleep disturbance, sexual dysfunctioning and weight gain. Therefore, it represents an unmet need to come up with antidepressant drugs of greater efficacy and improved tolerability as such treatments could lead to more complete remission in more patients. PMID- 18428080 TI - The lactate provocation test to investigate the relationships between panic attacks and psychosis: a report of two cases. AB - It was suspected that the delusional convictions of bewitchment and devil persecution of two female patients (41 and 40-years-old) could be the consequence of an erroneous interpretation of the sensations induced by panic attacks, as several authors have previously suggested. The interest of this case report stems from the manner in which we tested our clinical hypothesis. The patients agreed to the use of a lactate provocation test in double-blind, placebo-controlled conditions during four randomized sessions on consecutive days (two with lactate and two with placebo). The results for patient A strongly supported our hypothesis: patient A developed two full-blown panic attacks during the active lactate sessions, whereas patient B developed one subthreshold and one moderate panic attack during the active lactate sessions. The results of these investigations led to a more specific psychotherapeutic approach for patient A. PMID- 18428081 TI - Jacksonian seizure in a manic patient treated with rTMS. AB - This is a report of a jacksonian seizure in a non-psychotic manic patient who was treated with right prefrontal high frequency rTMS concomitant to pharmacotherapy. PMID- 18428082 TI - Can we help one billion sick people? PMID- 18428083 TI - Genomics in drug discovery: the best things come to those who wait. AB - The year 2007 has been marked by the maturation of high-throughput technologies that combine automation and miniaturization to enable systematic surveys of genome sequence variation, gene expression and gene function. These technologies have the potential to affect drug discovery in many ways, from target identification and validation, to pinpointing the molecular variants that influence medicine response. In the current climate of declining pharmaceutical R&D productivity, these approaches offer hope, but a price tag is attached. This review covers exciting advances in the field of genomics, and discusses when to act on genomic data versus when to wait for further information. PMID- 18428084 TI - Bioinformatics and cheminformatics: where do the twain meet? AB - Bridging the domains of cheminformatics and bioinformatics in the post-genomic era requires the convergence of goals, tools, techniques and annotations. This article reviews recent research at the interface of the domains that shows evidence of this convergence. While graph theoretical representations have long been used to develop simple topological descriptions of molecules, graph theory based network concepts are also widely employed in systems biology. Shape and conformation are important for understanding intermolecular interactions, and several structure-based cheminformatic descriptors have been developed and applied to drug-like molecules and biomolecules. Data fusion methods and shared ontologies can also help integrate data from multiple sources in order to generate a holistic picture of the shared molecular informatics domain. PMID- 18428085 TI - Mathematical biodescriptors of proteomics maps: background and applications. AB - This article reviews recent developments in the formulation and application of biodescriptors to characterize proteomics maps. Such biodescriptors can be derived by applying techniques from discrete mathematics (graph theory, linear algebra and information theory). This review focuses on the development of biodescriptors for proteomics maps derived from 2D gel electrophoresis. Preliminary results demonstrated that such descriptors have a reasonable ability to differentiate between proteomics patterns that result from exposure to closely related individual chemicals and complex mixtures, such as the jet fuel JP-8. Further research is required to evaluate the utility of these proteomics-based biodescriptors for drug discovery and predictive toxicology. PMID- 18428086 TI - Which aspects of HTS are empirically correlated with downstream success? AB - High-throughput screening (HTS) is a well-established hit-finding approach used in the pharmaceutical industry. In this article, recent experience at Novartis with respect to factors influencing the success of HTS campaigns is discussed. An inherent measure of HTS quality could be defined by the assay Z and Z' factors, the number of hits and their biological potencies; however, such measures of quality do not always correlate with the advancement of hits to the later stages of drug discovery. Also, for many target classes, such as kinases, it is easy to identify hits, but, as a result of selectivity, intellectual property and other issues, the projects do not result in lead declarations. In this article, HTS success is defined as the fraction of HTS campaigns that advance into the later stages of drug discovery, and the major influencing factors are examined. Interestingly, screening compounds in individual wells or in mixtures did not have a major impact on the HTS success and, equally interesting, there was no difference in the progression rates of biochemical and cell-based assays. Particular target types, assay technologies, structure-activity relationships and powder availability had a much greater impact on success as defined above. In addition, significant mutual dependencies can be observed - while one assay format works well with one target type, this situation might be completely reversed for a combination of the same readout technology with a different target type. The results and opinions presented here should be regarded as groundwork, and a plethora of factors that influence the fate of a project, such as biophysical measurements, chemical attractiveness of the hits, strategic reasons and safety pharmacology, are not covered here. Nonetheless, it is hoped that this information will be used industry-wide to improve success rates in terms of hits progressing into exploratory chemistry and beyond. The support that can be obtained from new in silico approaches to phase transitions are also described, along with the gaps they are designed to fill. PMID- 18428087 TI - Prioritizing hits from phenotypic high-content screens. AB - In the past decade, advances in the field of high-content screening (HCS) have provided researchers with a powerful new screening tool to observe treatment effects on multiple experimental parameters. While extremely useful, HCS has resulted in the collection of large datasets of increased complexity that require intensive analysis. Recently, approaches have been developed to analyze multi parametric HCS data more completely and, when used in conjunction with RNA interference, target-based biochemistry and structural analysis, these approaches have begun to unlock the potential of this screening format in aiding drug discovery. This review illustrates how the combination of these technologies has been used to successfully drive the drug discovery process. PMID- 18428088 TI - The synergy between combinatorial chemistry and high-throughput screening. AB - Despite the initial promise of combinatorial chemistry, particularly large library combinatorial chemistry, to greatly accelerate drug discovery, this approach has not been fully utilized as a means to build the compound collections of pharmaceutical and biotechnology companies. This review highlights some of the strengths of large library combinatorial chemistry as a means of generating molecules for lead discovery, such as providing rich and robust structure activity relationships around each hit series. The challenges and concepts emerging from traditional high-throughput screening and fragment-based drug design, how these methods influence the design of large combinatorial libraries and the interpretation of the ensuing high-throughput screening data are also highlighted. PMID- 18428089 TI - The use of protein-ligand interaction fingerprints in docking. AB - The use of structure-based virtual screening to predict small-molecule binding in a target active site is an increasingly popular approach in drug discovery programs. As the number of structures of protein-ligand complexes in public and proprietary databases grow, it is important to incorporate prior structural knowledge of ligand binding into virtual screening experiments. The structural interaction fingerprint (SIFt) approach aims to capture a 1D representation of the interactions between ligand and protein either in complexes of known structure or in docked poses. This review describes recent developments in the use of the SIFt in rescoring docked ligand poses in virtual screening. PMID- 18428090 TI - Recent developments in de novo design and scaffold hopping. AB - This review covers the developments in the fields of de novo ligand design and scaffold hopping since 2006. De novo ligand design was introduced in 1991 as a purely structure-based method to suggest ligands for synthesis and was later augmented by ligand-based approaches. Both structure-based and ligand-based methods identify pharmacophores, as well as shape constraints, and subsequently match these with complementary features embedded into small-molecule topologies. Recently, significant attention has been paid to de novo ligand design in combination with biophysical fragment screening and X-ray structure elucidation. Scaffold hopping has evolved from a niche application of de novo design into a rapidly expanding suite of different software tools, which are used extensively in the pharmaceutical industry. PMID- 18428091 TI - Recent trends in library design: 'rational design' revisited. AB - Diversity has historically played a critical role in the design of combinatorial libraries, screening sets and corporate collections for lead discovery. Large library design dominated the field of lead discovery in the 1990s, with design methods ranging from arbitrary and property-based reagent selection to product based approaches. Over time, however, there has been a downward trend in library size as the genomics revolution and the increasing availability of target protein structures from X-ray crystallography and homology modeling have increased the volume of information concerning desired targets. Concurrently, computing grids and CPU clusters have facilitated the development of structure-based tools that are able to screen hundreds of thousands of molecules. Smaller, 'smarter' combinatorial and focused parallel libraries have replaced the unfocused large libraries in the drug design paradigm of the 21st century. While diversity continues to play a role in lead discovery, the focus of current library design methods has shifted to scaffold design and bio-isostere searching, with a greatly needed emphasis on synthetic feasibility. PMID- 18428092 TI - Scientific workflows as productivity tools for drug discovery. AB - Large pharmaceutical companies annually invest tens to hundreds of millions of US dollars in research informatics to support their early drug discovery processes. Traditionally, most of these investments are designed to increase the efficiency of drug discovery. The introduction of do-it-yourself scientific workflow platforms has enabled research informatics organizations to shift their efforts toward scientific innovation, ultimately resulting in a possible increase in return on their investments. Unlike the handling of most scientific data and application integration approaches, researchers apply scientific workflows to in silico experimentation and exploration, leading to scientific discoveries that lie beyond automation and integration. This review highlights some key requirements for scientific workflow environments in the pharmaceutical industry that are necessary for increasing research productivity. Examples of the application of scientific workflows in research and a summary of recent platform advances are also provided. PMID- 18428093 TI - Software for rapid prototyping in the pharmaceutical and biotechnology industries. AB - The automation of drug discovery methods continues to develop, especially techniques that process information, represent workflow and facilitate decision making. The magnitude of data and the plethora of questions in pharmaceutical and biotechnology research give rise to the need for rapid prototyping software. This review describes the advantages and disadvantages of three solutions: Competitive Workflow, Taverna and Pipeline Pilot. Each of these systems processes large amounts of data, integrates diverse systems and assists novice programmers and human experts in critical decision-making steps. PMID- 18428094 TI - Public chemical compound databases. AB - The internet has rapidly become the first port of call for all information searches. The increasing array of chemistry-related resources that are now available provides chemists with a direct path to the information that was previously accessed via library services and was limited by commercial and costly resources. The diversity of the information that can be accessed online is expanding at a dramatic rate, and the support for publicly available resources offers significant opportunities in terms of the benefits to science and society. While the data online do not generally meet the quality standards of manually curated sources, there are efforts underway to gather scientists together and 'crowdsource' an improvement in the quality of the available data. This review discusses the types of public compound databases that are available online and provides a series of examples. Focus is also given to the benefits and disruptions associated with the increased availability of such data and the integration of technologies to data mine this information. PMID- 18428095 TI - [The adherence of clinical management in a randomized controlled trial. Results from a psychotherapy study]. AB - CM is increasingly used as a control condition in depression research. In the present study, the adherence of standardized CM sessions and their influence on outcome in depressed inpatients was investigated for the first time. In a randomized controlled trial, 43 inpatients with a diagnosis of Major Depression received medication treatment plus three-weekly CM sessions for 5 weeks. The 17 item version of the Hamilton Depression Rating Scale was the outcome measure. A total of 167 randomly selected audiotapes of CM sessions were evaluated using an adherence scale. The CM sessions delivered by psychiatric residents showed overall a high quality and included extensive support by the physician. The length of sessions was extended. The purity of sessions was associated with higher response rates. A diagnosis of Axis II-disorders or -traits and the clinical experience of the physician in years were predictors for less adherend treatments. However, both of them did not function as moderator variables. PMID- 18428096 TI - [Attachment style and outcome in short-term psychodynamic psychotherapy for children and adolescents]. AB - In several studies, the association between attachment style and psychotherapy outcome has been shown for adults. The aim of the present study was to examine the association between attachment style and outcome of short-term psychodynamic psychotherapy for children and adolescents. 71 children and adolescents with a mental disorder receiving out-patient short-term psychodynamic psychotherapy were studied. Attachment style was assessed with the Heidelberg Attachment Style Rating for Children and Adolescents (HASR-CA). Therapy outcome was measured using the Severity of Impairment Score for Children and Adolescents (SIS-CA). It could be shown that good outcome in therapy is significantly more frequently associated with a secure attachment style than with an insecure attachment style. No significant changes in attachment security were observed during treatment. Thus, attachment style is a good predictor of a successful short-term psychodynamic psychotherapy for children and adolescents and therefore is an important criterion for the differential indication for psychodynamic therapies. PMID- 18428097 TI - [Effectiveness of out-patient psychotherapy--a multimethod study in the German speaking part of Switzerland]. AB - The present paper describes an effectiveness-study to analyze the effectiveness of psychotherapeutic treatments. In the documentation phase of three years 24 therapists documented 93 psychotherapies. The patients represent a general population of health care takers. The treatments have an average duration of 45 appointments over 62 weeks. Good results are being achieved within this short period of treatment. The reduction of symptoms (global severity index GSI of SCL 90-R) reaches a high effectsize; regarding their symptoms 46 % of the patients can be considered as cured and 74 % as cured or improved. PMID- 18428098 TI - [Mourning, coping and subjective attribution after early miscarriage]. AB - A few weeks after miscarriage 232 women were questioned with standardized questionnaires and symptom scales about the way in which they coped with an early miscarriage (up to the 16th week of gestation) about mourning processes and subjective attribution as well as anamnestic and psychosocial characteristics. Seven and fourteen months after pregnancy loss and if necessary in the first trimester of a following pregnancy the psychological symptoms of the women (fear and depressive symptoms) were evaluated. A few weeks after miscarriage we found with main component analysis three patterns of coping. The pattern of "depressive coping" could be diagnosed as a predictor of increased depression seven months after the miscarriage and in a new pregnancy. Depressive pathology in a new pregnancy of the women with early pregnancy loss can be predicted also over the pattern of "anxious grieving". On the other hand, the pattern of "active coping" is associated with lower state anxiety in a new pregnancy. Lack of perceived partner support and past divorces or separations are risk factors for increased fears in a new pregnancy. These results suggest that women who have had an early miscarriage especially if they have maladaptive coping patterns and psychosocial risk factor for maladjustment need prevention of psychological disturbances with an integrated psychosomatic concept. PMID- 18428099 TI - [Prevalence of a pathological DGER (duodeno-gastric-oesophageal reflux) in patients with clinical symptoms of reflux disease]. AB - BACKGROUND: According to recent studies DGER (duodeno-gastric-oesophageal reflux) is considered as an independent risk factor for the development of reflux esophagitis and the Barrett metaplasia. The Bilitec 2000 allows a qualitative and quantitative measurement of DGER in patients with symptoms of reflux disease. The aim of the present study was to investigate the prevalence of DGER in patients with reflux symptoms. METHODS: 146 patients with symptoms of gastro-oesophageal reflux disease were enrolled in this study. Patients underwent upper gastrointestinal endoscopy, oesophageal manometry and simultaneous 24 h oesophageal pH and bilirubin monitoring. The presence of pathological DGER and its relations to the symptom pattern, distal oesophageal acid exposure and endoscopic findings were analysed. RESULTS: In 74 out of 146 patients (51 %, 39 men, 34 women) a DGER could be detected. Twenty-eight (32 %) of these patients suffered from an isolated DGER, while 46 (32 %) had a combined acid and DGER reflux. An isolated acid reflux was found in additional 28 (19 %) patients. The degrees of both acid and DGER were significantly higher in those patients with oesophageal lesions. CONCLUSIONS: 1. There is a high prevalence of DGER in patients with the clinical symptoms of a reflux disease. 2. The combined measurement of acid reflux and DGER helps to better define the cause of reflux symptoms. 3. In analogy to the acid reflux DGER increases with the gravity of oesophageal lesions. PMID- 18428100 TI - [A pharmacoepidemiological study of antiepileptic drug consumption (1992-2004)]. AB - INTRODUCTION: Epilepsy and the epileptic syndromes are common neurological diseases and represent an important public health problem that has given rise to marked social and healthcare concerns. AIM: To analyse the changes in the consumption of antiepileptic drugs in the Basque Country Autonomous Community over a 13 year period (1992-2004). MATERIALS AND METHODS: The consumption of the N03 subgroup has been studied using data from the ECOM database of the Spanish Ministry of Health and Consumer Affairs, which records the number of drug packets dispensed on National Health Service prescription. The results are expressed as defined daily doses per 1,000 inhabitants per day (DID). RESULTS: A significant increase is observed in the consumption of antiepileptic drugs over the study period (5.53-9 DID). The most widely used drugs were phenobarbital, carbamazepine, valproic acid and phenytoin and, in recent years, the extensive use of gabapentin is of particular note. CONCLUSION: Epidemiological studies on epilepsy are difficult to perform due to a number of methodological problems which are caused fundamentally by the heterogeneity of the disease. Pharmacoepidemiological studies constitute a simple, effective, low-cost tool for estimating the prevalence of the disease, and furthermore enumerates the drugs used, thus contributing to a rational use of these drugs. PMID- 18428101 TI - [Development and validation of a short battery of tests for the assessment of aphasia: 'bedside assessment of language'. Its use in a rehabilitation centre]. AB - INTRODUCTION: A formal and complete language assessment requires too much time and also tires the patient out during the first recovery stages after brain injury. The aim of this paper is to introduce a bedside screening, short and sensitive to diagnose aphasias in patients with brain injury. PATIENTS AND METHODS: The results of the Bedside Assessment of Language (BAL) was studied in 121 aphasic patients and in 77 dysarthric patients after brain injury. The five linguistic areas assessed in this short battery are: spontaneous language, comprehension, repetition, writing and reading. Each area has a maximum of 5 points, and the test has a total score of 25. Statistics tests used were: median test, Kruskal-Wallis test, Fisher's exact test, receiver operating characteristics curves and correlation coefficient. RESULTS: The BAL showed an excellent internal consistency reliability (Cronbach's alpha coefficient: 0.9419); sensitivity was of 79.3% and specificity of 84.4%. CONCLUSIONS: These results indicate that the BAL has been a potentially useful tool for the diagnose of the type of aphasia and it has been sensitive to evolutionary changes in sub acute stages of language disorders. PMID- 18428103 TI - [Cognitive effects of combined memory and psychomotor training in elderly adults]. AB - INTRODUCTION: As part of the normal ageing process, cognitive deterioration clearly takes place in memory, attention and the information processing speed (IPS). Among the intervention strategies commonly used, combined programmes like those involving memory and psychomotor skills have the greatest beneficial effects on cognition in the short and long term. AIM: To evaluate the effects of a combined memory and psychomotor training programme entitled 'Independence in Older age' (SIMA) on the cognitive performance of a population sample of healthy elderly adults. SUBJECTS AND METHODS: A cross-sectional study was conducted on 95 elderly adults (49 in the intervention group and 46 in the control group) before and after a 20-session training programme. Both intergroup (intervention group versus control group) and intragroup (each group individually) cognitive performance was analysed. The two groups were paired by age, sex and schooling. RESULTS: The intergroup analysis following the training showed statistically and clinically significant differences in two IPS variables: conflict and interference in the colour-word test, which had a size effect of -1.31 and -1.38, respectively; the performance of the group of trained elderly adults was better than that of the controls. Likewise, in the intragroup analysis significant differences were found in these two variables in the group that received the intervention, with a size effect of -1.27 and -1.15. CONCLUSION: Findings show positive effects of combined SIMA memory and psychomotor training, especially in selective attention and the IPS with a healthy elderly adult population. PMID- 18428104 TI - [Miller Fisher syndrome as the presenting symptom of Wernicke's encephalopathy]. AB - INTRODUCTION: Wernicke's encephalopathy is an acute neuropsychiatric syndrome resulting from a thiamine deficit, which is defined by the characteristic triad of confusion, ophthalmoparesis and ataxia, although rare presentations have been reported that delay its diagnosis. Miller Fisher syndrome is characterised by the triad ophthalmoparesis, ataxia and areflexia and is considered to be a variant of Guillain-Barre syndrome; its differential diagnosis includes Wernicke's encephalopathy. CASE REPORT: A 75-year-old female with chronic digestive disorders, who developed an acute picture of bilateral internuclear ophthalmoplegia, ataxia and areflexia, with proteinocytologic dissociation in cerebrospinal fluid; accordingly, an initial diagnosis of Miller Fisher syndrome was proposed. Results of the neurophysiological studies were normal; anti-GQ1b antibodies were negative; and magnetic resonance imaging of the brain suggested Wernicke's encephalopathy. The response to thiamine was spectacular. CONCLUSIONS: The similarities in the distribution of the lesions of the two conditions, in the signs and symptoms and the lab findings, as well as the influence of certain misleading factors (hyponatremia, advanced age), went to make up a typical syndrome that favoured a wrong presumptive aetiological diagnosis. This was corrected at an early stage, however, in light of the results of certain diagnostic tests and after observing the therapeutic response. In addition to being an atypical presentation for Wernicke's encephalopathy, this case highlights the fact that for there to be an agreement between the syndromic and aetiological diagnoses it is necessary to carry out a correct differential diagnosis based on details from the patient's history, on appropriate complementary tests and on the follow-up study of how the patients progress, even when we come across typical syndromes that are usually related to a predominant aetiopathogenesis. PMID- 18428105 TI - [Ziprasidone in Parkinsonian dopamine psychosis]. AB - INTRODUCTION: The dopamine psychosis that appears in Parkinson's disease (PDP) is a complication that is often related with frequent intake of antiparkinsonian agents, especially levodopa and dopamine agonists. Morbidity and the risk of being institutionalised are increased in such patients and therapeutic management is difficult. Its treatment is based on reducing the intake of antiparkinsonian drugs or the use of atypical antipsychotics, due to the fact that they have scarce or no motor effects (at least in theory). In this work, we examine the role that ziprasidone (ZPS) can play in PDP. DEVELOPMENT: We reviewed the studies in which ZPS was used to treat PDP that have appeared in the literature. To date, very few studies have been conducted and only a small number of patients were included in each case. Nevertheless, it seems that, at doses below those used in schizophrenia (20-80 mg/day), ZPS is an effective antipsychotic. In general, the extrapyramidal side effects that do appear are infrequent and mild. Pathological laughter was detected in some patients. The drug was well tolerated and safe. CONCLUSIONS: ZPS seems to be an effective antipsychotic for PDP, with scarce or no extrapyramidal side effects; it is also safe and well tolerated. In view of the scarcity of studies and patients, however, further research is needed (if possible, by means of double-blind randomised trials) to be able to assess the true role of ZPS in PDP. PMID- 18428102 TI - [Treatment pattern of Alzheimer's disease with cholinesterase inhibitors (TRAIN study)]. AB - AIMS: To describe the relation between the level of cognitive impairment in Alzheimer's disease and the use of cholinesterase inhibitors (ChEIs) in neurology, geriatric and psychiatric units, and to establish the clinical profile of these patients. PATIENTS AND METHODS: An epidemiological, multi-centre, cross sectional study was conducted. Subjects included in the study were consecutive outpatients diagnosed with Alzheimer's disease, in accordance with the fourth edition of the Diagnostic and Statistical Manual of Mental Disorders, and who had been treated with rivastigmine, donepezil or galantamine, either on its own or in association with memantine in the last six months. The recruitment period lasted three months. In a single visit, researchers determined the medication that was used, the dose, the mini-mental test, the overall clinical impression-overall improvement and the overall clinical impression-severity of the disease. A total of 1940 patients were selected from neurology, psychiatric and geriatric services all over the country. Possible differences in the habits of different specialists as regards prescribing were analysed, together with the relation between cognitive impairment and the type of medication employed. RESULTS: The mean age of the patients was 77 +/- 6.6 years, 62% of whom were females; the mean score on the mini-mental test was 17.4 +/- 5.5. The mini-mental score was similar in patients treated with rivastigmine (18.02 +/- 5.23), donepezil (17.08 +/- 5.54) or galantamine (17.34 +/- 5.38). In patients who were treated with memantine in association with a ChEI, the mini-mental score was significantly lower (11.44 +/- 5.68) (p < 0.0001). The doses of the different ChEIs used by the specialists were similar. A higher percentage of patients had maximum doses of donepezil (81%) than in the cases of rivastigmine (43%) and galantamine (67%). CONCLUSIONS: The different specialists involved (neurologists, geriatricians and psychiatrists) displayed similar habits regarding the utilisation of ChEIs to treat Alzheimer's disease. There was no relation between the degree of impairment and the drug chosen, except in the case of memantine. PMID- 18428106 TI - [The impact of lesions in the right hemisphere on linguistic skills: theoretical and clinical perspectives]. AB - INTRODUCTION: A lesion of the right hemisphere of right-handers can result in verbal communication impairments. The recent development of theoretical frameworks with regard to discourse and pragmatic abilities, among others, now allows us to recognize and describe these impairments. AIM: To offer an overview of the verbal communication deficits that can be found in right-hemisphere damaged individuals. These deficits can interfere, at different levels, with prosody, the semantic processing of words and discourse and pragmatic abilities. DEVELOPMENT: Such impairments appear to be present in about half of right hemisphere-damaged patients and, when present, can result in different clinical profiles. These deficits raise the question of their labeling and their relationship with aphasia. CONCLUSIONS: Given the evolution of the concept of language and the universal definition of aphasia, it is proposed that these deficits correspond to another manifestation of aphasia, thus challenging the idea that they are of a 'non-aphasic' nature. PMID- 18428107 TI - [The mad and the demented in the literary works of Cervantes: on Cervantes' sources of medical information about neuropsychiatry]. AB - INTRODUCTION: Throughout the writings of Cervantes, the lunatic or the madman (which allows for certain neurological pathologies, such as epilepsy) is a figure that appears over and over again, as a literary strategy that the author uses to express his own particular social criticism. DEVELOPMENT: This literary characterisation of insanity is endowed with a number of clinical connotations that suggest that Cervantes was quite familiar with certain elements of medicine; although such knowledge may well have come from relatives and friends, he also showed a personal interest in the mentally ill (there is evidence that he visited the Hospital de Inocentes--a mental asylum--in Seville). Likewise, it seems obvious that Cervantes was also familiar with several medical treatises concerning the neurosciences, which were very much in vogue in Spain in the late Renaissance period. In fact, his personal library included two works that, in addition to serving to inspire the Alcala-born author on some points, are cited almost literally in some of his novels. These references are The Examination of Men's Wits, written by Juan Huarte de San Juan in 1575, and the annotated Spanish version of Dioscorides (On medical matters and mortal poisons) by Andres Laguna, from 1555. Similarly, the clinical description of the main character in the novela ejemplar Vidriera, the Lawyer seems to have been taken from the medical tract Dignotio et cura affectuum melancholicorum (1569) by Alfonso de Santa Cruz. Lastly, it has also been suggested that, in the construction of his literature, Cervantes drew on the humanistic approaches advocated by Erasmus of Rotterdam in his In Praise of Folly (1509). CONCLUSIONS: These four works may have served as a source of medical information that allowed Cervantes to develop some of his characters and to write his accurate comments on therapeutic matters. PMID- 18428109 TI - [Trigeminal involvement in SUNCT syndrome]. PMID- 18428111 TI - [SIP30-AI: reflections]. PMID- 18428113 TI - Directed evolution of LuxI for enhanced OHHL production. AB - Quorum sensing is a common mechanism used by bacteria to coordinate population behavior, and is involved in a variety of biological processes, such as bioluminescence, virulence factor synthesis, antibiotic production, and biofilm formation. To engineer the LuxI enzyme of the LuxI-LuxR quorum-sensing system, we developed a high throughput genetic selection to identify LuxI mutants with improved OHHL (3-oxo-hexanoyl homoserine lactone) synthesis in E. coli. Using this genetic selection, we created LuxI mutants with improved OHHL synthesis rates and yields through directed evolution, identifying three LuxI mutants after two generations. An in vivo semi-quantitative method allowed for verification of the genetic screen and OHHL yields were quantified using HPLC-MS/MS, revealing an 80-fold increase in a mutant culture compared to the wildtype culture. In addition to OHHL, the yields of C6HSL (hexanoyl homoserine lactone) and C8HSL (octanoyl homoserine lactone) were also improved, and a slight change in substrate specificity towards C6HSL production was observed. Based on alignment with the crystal structure of EsaI, a homolog of LuxI, two mutations are most likely involved in enhancing the interactions between the enzyme and the substrates. The high throughput genetic selection and the semi-quantitative method can be conveniently modified for the directed evolution of LuxI homologs. The identification of these LuxI mutants has implications in synthetic biology, where they can be used for the construction of artificial genetic circuits. In addition, development of drugs that specifically target quorum sensing to attenuate the pathogenesis of gram-negative infectious bacteria might also benefit from the insights into the molecular mechanism of quorum sensing revealed by the amino acid substitutions. PMID- 18428114 TI - Polymeric particles conjugated with a ligand to VCAM-1 exhibit selective, avid, and focal adhesion to sites of atherosclerosis. AB - The increased expression of VCAM-1 on endothelial segments within plaque regions could be used as a target to deliver polymeric drug carriers selectively to sites of atherosclerosis. We probed the hypothesis that polymeric particles conjugated with a ligand for VCAM-1 exhibit selective and avid adhesion to sites of atherosclerosis. Particles made from polystyrene or the biodegradable polymer poly(sebacic acid)-block-polyethylene glycol (PSA-PEG) were conjugated with an antibody to VCAM-1 (alpha-VCAM-1) or IgG (negative control). The particles were injected into the jugular vein of ApoE(-/-) (a murine model of atherosclerosis) or wild type mice and their adhesion to the aorta determined. alpha-VCAM-1 particles exhibited significantly greater adhesion to ApoE(-/-) mouse aorta [32 +/- 5 (mean +/- SEM) particles/mm(2) for polystyrene particles and 31 +/- 7 particles/mm(2) for PSA-PEG particles] compared to the level of adhesion to wild type mouse aorta (18 +/- 1 particles/mm(2) for polystyrene particles and 6 +/- 1 particles/mm(2) for PSA-PEG particles). Within ApoE(-/-) mice, the alpha-VCAM-1 particles exhibited significantly greater adhesion to the aorta (32 +/- 5 particles/mm(2) for polystyrene particles and 31 +/- 7 particles/mm(2) for PSA PEG particles) compared to the adhesion of IgG particles (1 +/- 1 particles/mm(2) for polystyrene particles and 2 +/- 1 particles/mm(2) for PSA-PEG particles). Detailed analysis of the adhesion revealed that alpha-VCAM-1 particles exhibited focal adhesion to plaque regions, in particular the periphery of the plaques, within the ApoE(-/-) mouse aorta. Combined the data demonstrate that polymeric particles conjugated with a ligand to VCAM-1 exhibit selective, avid and focal adhesion to sites of atherosclerosis providing strong evidence that VCAM-1 ligand bearing polymeric particles could be used for targeting drugs selectively to atherosclerotic tissue. PMID- 18428115 TI - Conformational analysis of molecular machines: internal rotation and enantiomerization in triptycyl[3]helicene. PMID- 18428116 TI - Nonpeptidic vinyl and allyl phosphonates as falcipain-2 inhibitors. PMID- 18428117 TI - Neurotrophic factor-related gene polymorphisms and adult attention deficit hyperactivity disorder (ADHD) score in a high-risk male population. AB - Adult attention deficit hyperactivity disorder (ADHD) is a widely under-reported but nevertheless common condition with a clear heritable component. Several genes have been proposed to play a role in the childhood onset of this neurodevelopmental disorder; however, association studies of persistence of ADHD into adulthood have rarely been performed. Neurotrophic factors (NTFs) are known to be involved in several aspects of neuronal development and neural plasticity in adults. They have also been linked, particularly through brain-derived neurotrophic factor (BDNF) interaction with dopamine transport, to the pathophysiology of ADHD. This study compares the genotypes of six different single nucleotide polymorphisms of genes within the neurotrophin system and their possible association with adult ADHD score in 143 high-risk male subjects referred to a forensic psychiatric unit. The genes included NTF3, NTRK2 (TrkB), NTRK3 (TrkC), BDNF, and p75(NTR). While none of the SNPs showed significant association with ADHD symptoms, one polymorphism within the exon of NTF3 (rs6332) showed a trend toward an association between the A-allele and increased scores using both the retrospective childhood analysis Wender-Utah Rating Scale (WURS-k) (P = 0.05) and the adult ADHD assessment Wender-Reimherr interview (P = 0.03). This SNP is a silent mutation which might be in linkage disequilibrium with a functional risk variant for ADHD. As the association was only suggestive, however, this finding needs replication in a larger study with higher power. PMID- 18428118 TI - Literature review: January-March 2008. PMID- 18428119 TI - Self-reported early trauma as a predictor of adult personality: a study in a military sample. AB - Exposure to early trauma has frequently been linked to adult psychopathology, including personality disorders. This cross-sectional study explored the relationship between personality and retrospectively rated levels of early trauma in 242 soldiers. Multiple regression analyses showed a significant relationship between early trauma and adult personality as exposure to emotional trauma predicted levels of self-directedness and cooperativeness on the Temperament and Character Inventory (TCI; C. R. Cloninger, T. R. Przybeck, D. M. Svrakic, & R. D. Wetzel, 1994). Overall, these results suggest that early emotional trauma may be related to personality dimensions associated with poor impulse control and interpersonal behavior. These results are noteworthy considering that they were obtained in a healthy sample. They show that early trauma may become ingrained in personality and hamper the potential to effectively engage in social interactions, increasing the risk of emotional and cognitive problems. PMID- 18428120 TI - Asymptomatic reactivation and shed of infectious varicella zoster virus in astronauts. AB - Varicella zoster virus (VZV) causes varicella (chickenpox), after which virus becomes latent in ganglia along the entire neuraxis. Virus reactivation produces zoster (shingles). Infectious VZV is found in vesicles of patients with zoster and varicella, but virus shed in the absence of disease has not been documented. VZV DNA was previously detected in saliva of astronauts during and after spaceflight, a uniquely stressful environment in which cell mediated immunity (CMI) is temporally dampened. The decline in CMI to VZV associated with zoster led to the hypothesis that infectious VZV would also be present in the saliva of astronauts subjected to stress of spaceflight. Herein, not only was the detection of salivary VZV DNA associated with spaceflight validated, but also infectious virus was detected in saliva from 2 of 3 astronauts. This is the first demonstration of shed of infectious VZV in the absence of disease. PMID- 18428121 TI - Prevalence of VP4 and VP7 genotypes of human rotavirus in Ecuadorian children with acute diarrhea. AB - The objective of the present study was to determine rotavirus etiology and prevalence of the different rotavirus serotypes in Ecuadorian children younger than 5 years of age with gastroenteritis. Children (729) less than 5 years of age with acute diarrhea from either public or private primary health care centers in 10 different provinces of Ecuador, between March 2006 and August 2006 were included in the study. Rotavirus infection was diagnosed using a commercial immunoenzymatic test. Rotavirus isolated from stool samples was genotyped. Rotavirus was detected in the feces of 269 of the 729 children (37%) with diarrhea. The most prevalent G genotypes were G9 (46.1%) and G2 (27.2%), while the predominant P genotypes were P[8] (57%) and P[4] (29.5%). Among the single infections, the predominant P/G combinations were: P[8]G9 (56.9%) and P[4]G2 (32.6%). The present countrywide survey is one of the major studies for one single season in Latin America and the first in its class in Ecuador. The value of expanding laboratory capability throughout Latin America in order to monitor rotavirus strains over time, with special attention directed at those strains obtained from children who experience vaccine failure, is critical. Only continuous monitoring of rotavirus disease burden and genotype surveillance will provide this information. PMID- 18428122 TI - Monitoring of adenovirus from conjunctival scrapings in Japan during 2005--2006. AB - A total of 189 conjunctival scrapings were collected from patients in Tokyo, Japan by monitoring adenovirus infection in community-based clinics during 2005 and 2006. Of the 189 samples, 155 (82%) had adenoviruses detected by polymerase chain reaction (PCR). The serotypes were determined by PCR-restriction fragment length polymorphism (PCR-RFLP) analysis, using a combination of endonucleases, such as HhaI, AluI, and HaeIII, and neutralization tests (NTs). PCR-RFLP identified five serotypes: serotype 3: 16.8%, serotype 4: 9.7%, serotype 8: 34.8%, serotype 11: 23.2%, and serotype 37: 15.5%. Adenovirus 8 was the most common serotype identified. A subset consisting of 25 isolates identified as adenovirus 8 from this study plus 25 isolates from Kawasaki were analyzed using PCR sequencing of the hexon gene. Compared with prototype adenovirus serotype 8 and serotype 9 derived in Tokyo and Kawasaki, these isolates shared 61.7-62.8% and 80.5-82.7% amino acid homology, respectively, suggesting that a variant adenovirus serotype 8 was involved in this outbreak, and is different from the prototype adenovirus 8 virus. This variant had not been detected in Japan prior to 1996 and appears to be the most common adenovirus type 8 involved in cases of epidemic keratoconjunctivitis in Japan at present. PMID- 18428123 TI - Crimean-Congo hemorrhagic fever in Pakistan. AB - Crimean-Congo virus, the causative agent of Crimean-Congo Virus Fever (CCVF) is endemic in Pakistan. Cases are documented sporadically ever year, mostly at and around the time of Eid-ul-Adha, an Islamic festival, celebrated on day 10 through 13 of the 12th month of each lunar calendar year. At this time of the year in Pakistan, livestock are brought down to the urban areas from the rural parts of the country. Animals are housed in open spaces and private houses until they are slaughtered during the 3 days of Eid-ul-Adha. This allows the CCHF virus, which is carried by a tick that inhabits the animal hide, to be transmitted through unprotected contact with live animals as well as through contact with animal blood subsequent to its slaughter. In this report, a typical case of CCVF is described that was encountered in Rawalpindi, Pakistan. A number of issues pertaining to the management of recurrent outbreaks of CCVF in the country are discussed. PMID- 18428124 TI - Human cytomegalovirus and Epstein-Barr virus in apical and marginal periodontitis: a role in pathology? AB - Periodontitis is presumably caused by bacterial infection, but it has been shown recently that affected tissue often contains human cytomegalovirus (HCMV) and Epstein-Barr virus (EBV). The present study was initiated to evaluate the role of these viruses in the pathogenesis of periodontitis. HCMV and EBV were quantified in 40 apical and 25 marginal periodontitis samples using real time PCR. In situ hybridization or immunohistochemistry was carried out on apical samples to detect viral presence within cells. A possible association with relevant bacteria was examined. Of the apical periodontitis samples, 50% contained EBV, while none contained HCMV. Of the marginal periodontitis samples, 40% were positive for EBV and 12% for HCMV. With one exception, however, the amount of virus was close to the detection limits. EBV was only detected in 1 out of 15 healthy periodontium samples. Immunohistochemistry and in situ hybridization were all negative. Significant associations were found between periodontal EBV and the presence of Aggregatibacter actinomycetemcomitans and Porphyromonas gingivalis. Although there was an obvious association of the virus with clinical samples, it seems unlikely that these viruses play a major role in the pathogenesis of periodontitis of the average patient. Their presence may reflect that the clinical samples contain more blood or saliva compared to controls, or an accumulation of lymphoid cells harboring virus in the inflamed tissue. PMID- 18428126 TI - DNA microarray technology for simultaneous detection and species identification of seven human herpes viruses. AB - The aim of the study was to develop a multiplex PCR-based DNA microarray technology for simultaneous detection and species identification of seven human herpes viruses, namely herpes simplex virus type 1, type 2 (HSV-1, HSV-2), varicella-zoster virus (VZV), Epstein-Barr virus (EBV), cytomegalovirus (CMV), and human herpes virus 6 (HHV-6A, HHV-6B), and to apply this technology to accurate diagnosis of herpesvirus-associated diseases. Primers and oligonucleotide probes were designed and synthesized based on the highly conserved regions of the DNA polymerase gene in human herpes viruses. DNA microarrays were made by printing the oligonucleotide probes onto special glass slides. After amplification and labeling with CY5, the PCR products were hybridized with the DNA microarrays and species identified. Sixty-one cerebrospinal fluid (CSF) and 132 blood specimens were analyzed by this technique, and the results were compared with those of TaqMan PCR. Several specimens were sequenced further after cloning. The PCR products of the seven human herpes viruses ranged from 224 to 252 bp, and could be species identified with DNA microarrays. The detection limits were 10(1) copies/microl for each virus. And the test showed no cross-reaction to DNA extracted from S. aureus, E. coli, hepatitis B virus, Cryptococcus neoformans, Candida albicans and human genome. Among 132 blood and 61 CSF specimens, 55 were tested positive for human herpes virus DNA. Compared with the results of TaqMan PCR, the sensitivity and specificity of the DNA microarray technology was 96.2% and 99.3%, respectively. This multiplex PCR-based DNA microarray technology, which is rapid, specific and sensitive, serves as an effective technique for simultaneous detection and species identification of seven human herpes viruses. PMID- 18428125 TI - Molecular epidemiological study of HEV-B enteroviruses involved in the increase in meningitis cases occurred in Spain during 2006. AB - Human enteroviruses are one of the main etiological agents of aseptic meningitis and other central nervous system infections, particularly the serotypes included in the enterovirus B species. Molecular methods have proved useful to identify serotypes in clinical samples, facilitating the epidemiological study of these viruses. In the spring of 2006, there was a significant increase in meningitis cases caused by enteroviruses in Spain. In the present study, 138 enteroviruses directly detected in clinical samples of patients with aseptic meningitis (n = 116) and other neurological pathologies (n = 22) received by the National Center for Microbiology during the year, were genotyped by amplification and sequencing part of the VP1 region and phylogenetic analysis. Echovirus 30 was the most frequent serotype, followed in decreasing order by echovirus 6, 9, 13, 18, enterovirus 75, coxsackievirus A9, echovirus 11, 14, 29, 4, and coxsackievirus B4 and B5. Phylogenetic analysis with all Spanish echovirus 30 strains detected in 2006 and other reported echovirus 30 sequences, demonstrated that Spanish strains formed a new lineage, different from others previously described. In conclusion, echovirus 30 is the most commonly reported enterovirus serotype associated with aseptic meningitis in Spain. Direct molecular typing of clinical samples also allows rapid identification of the serotypes involved in an epidemic alert and phylogenetic analysis in the 3'-VP1 region is useful to study viral epidemiology. PMID- 18428128 TI - Serologic and genetic characterization analyses of a highly pathogenic influenza virus (H5N1) isolated from an infected man in Shenzhen. AB - Highly pathogenic avian influenza (HPAI) H5N1 virus caused a wave of outbreaks in China during 2005--2006, resulting in a total of 20 cases of human infection in 14 provinces of China. On June16, 2006, a case of H5N1 human infection was confirmed in Shenzhen. The virus isolated from the patient, A/Guangdong/2/06, was characterized genetically and the relationship between the tracheal virus load and the antibody titer of the infected man was analyzed. Serological analysis confirmed that the patient's neutralizating antibodies had been generated 2 weeks after the onset of symptoms. The patient's serum antibodies could efficiently neutralize A/Guandong/2/06 infectivity in vitro. Phylogenetic analysis showed that the H5N1 virus of Shenzhen belonged to subclade 2.3.4, which contained viruses that were mainly responsible for the outbreaks in domestic poultry and in the cases of human infection in southern China. Homology and molecular characterization analysis revealed that all the segments of Shenzhen H5N1 virus still belonged to avian segments. Several specific amino acid residue mutations were detected. PMID- 18428127 TI - Genetic characterization of hantaviruses isolated from Guizhou, China: evidence for spillover and reassortment in nature. AB - To gain more insights into the epidemiology of hantaviruses in the Guizhou province, China, rodents were captured in Guizhou during the period from 2001 to 2003. In addition, serum sample was collected from one patient. Virus isolation was attempted from human serum and rodent samples. Four hantaviruses were isolated successfully in cell culture from one human, two A. agrarius, and one R. norvegicus. The nucleotide sequences for the entire S and M and partial L segment were determined from these four isolates as well as six viruses isolated in 1980s. Phylogenetic analysis revealed that the S segment from all isolates belong to the Hantaan virus (HTNV) clade, regardless of the sources from which they were derived. According to the S sequences, these viruses could be divided into three distinct phylogroups, showing geographical clustering. Analysis of the entire M and the partial L segment sequences demonstrated that 8 out of the 10 isolates belong to the HTNV clade. However, two isolates (CGRn8316 and CGRn9415) isolated from R. norvegicus belong to the Seoul virus (SEOV) clade. In addition, these two isolates were distinct from other known members of SEOV clade. Together, the data suggest that at least three groups of HTNV are co-circulating and one new variant of SEOV may be present in Guizhou. Our results also suggest that HTNV from A. agrarius spilled over to R. norvegicus and natural reassortment between HTNV and SEOV occurred during or after the spillover. PMID- 18428129 TI - Human influenza virus infection and apoptosis induction in human vascular endothelial cells. AB - Acute encephalopathy accompanying influenza virus infection results in brain and systemic organ failure mainly through vasogenic edema with high levels of inflammatory cytokines, such as blood tumor necrosis factor (TNF)-alpha and interleukin (IL)-6, as well as the cytochrome c apoptosis marker. A highly virulent strain of avian influenza virus causes fatal infection in chickens by infecting vascular endothelial cells in systemic organs, inducing apoptosis therein. To verify the possibility of apoptosis induction by human influenza virus in infected human vascular endothelial cells, purified influenza virus infected human umbilical vein endothelial cells (HUVECs) were examined using a tissue culture method. When pre-treated with TNF-alpha, influenza virus (Philippine strain, H3N2) promoted TNF-alpha induced apoptosis of HUVECs. Viral replication was confirmed in HUVECs infected with the Philippine strain in the absence of TNF-alpha by measurement of the amount of infective virus in the culture supernatant using the tissue culture infectious dose (TCID) method, immunohistochemistry and real-time PCR. The number of influenza virus genomes in the infected HUVECs at 24 hr post-infection increased about fivefold compared to that just after virus adsorption. Many TUNEL-positive influenza virus-infected HUVECs were observed using the TUNEL method. Furthermore, cleaved caspase 3 was also detected in influenza virus-infected cells by immunofluorescence staining. These results demonstrated that human influenza virus can infect and replicate in human vascular endothelial cells and induce apoptosis therein. PMID- 18428130 TI - Occurrence rate and genotype distribution of the JC virus (JCV) in a sample from the Polish population. AB - The JC virus (JCV) is a common human virus persisting in renal tissue. In immunocompromised individuals it may reactivate and cause progressive multifocal leukoencephalopathy (PML). JCV has also been implicated in cancerogenesis, leading to various brain tumors and cancers of gastrointestinal tract. In this study the JCV excretion in urine of 113 healthy Polish donors was analyzed. A 215 bp region of the viral gene coding for a major capsid protein VP1 was PCR amplified and detected in 52 individuals (46.0%). The occurrence rate increased with age and was highest in the group of over 60-year-old donors (63.6%). Sequence analysis of the VP1 gene fragment revealed the following distribution of JCV genotypes in the investigated group: 1A, 31 (59.6%); 1B, 13 (25.0%); 2A, 2 (3.8%); and 2C, 6 samples (11.5%). The frequency and distribution of the JCV genotypes in the Polish population resembles that in other European countries, with the most abundant genotype 1 (84.6%). However, while in those countries the second most frequent genotype was usually 4, in the investigated group genotype 4 was not detected. PMID- 18428131 TI - The course of fever following influenza virus infection in children treated with oseltamivir. AB - Although the effectiveness of oseltamivir against influenza virus infection is well known, there has been no report analyzing the detailed time course of fever following the drug treatment in children. Oseltamivir was prescribed for 4 days to every child with a positive result for rapid immunological test for influenza virus during 2002--2003, 2003--2004, and 2004--2005 epidemics. Only those who were 1-12 years of age and prescribed oseltamivir within 24 hr after the onset of fever were included in the analysis. The numbers of children with type A/H3N2 disease for the three seasons were 64, 77, and 33, and those with type B disease were 102, 4, and 86, for the respective seasons. The period until normalization of temperature was obtained from six-hourly recordings of body temperature. By multiple regression analysis, temperature periods were longer in type B than in type A/H3N2 disease, negatively associated with age, and positively with maximal body temperature (all: P < 0.001). The effectiveness of oseltamivir on body temperature in type B disease was less apparent in the 2004--2005 than in the 2002--2003 season, irrespective of age. No such between-season difference was observed for Type A/H3N2 disease. Frequencies of ineffective cases with biphasic fever (19.6% and 43.0% during 2002--2003 and 2004--2005 seasons) were significantly higher in type B than in type A/H3N disease (12.0% and 11.8%, respectively). The effectiveness of oseltamivir depends on a child's age, maximal body temperature and the virus type. This study confirmed recent reports indicating decreased effectiveness of oseltamivir against type B disease. PMID- 18428132 TI - Non-structural protein NSP2 induces heterotypic antibody responses during primary rotavirus infection and reinfection in children. AB - Rotaviruses are the single most important causes of severe acute diarrhoea in children worldwide. Despite success in developing vaccines, there is still a lack of knowledge about many components of the immune response, particularly those to non-structural proteins. This study established radioimmunoprecipitation (RIP) assays using labeled G1P[8], G2P[4], and G4P[6] human rotaviruses to examine the spectrum and duration of rotavirus antibodies in sera collected sequentially for 18-36 months from 27 children after hospitalization for primary rotavirus gastroenteritis. Five children experienced rotavirus re-infections. Primary responses detected to non-structural protein NSP2 declined to baseline after 100 150 days. Responses were heterotypic between NSP2 of G1P[8] and G4P[8] rotaviruses. Re-infections after 465-786 days boosted antibody levels to NSP2of both serotypes, together with the appearance of anti-NSP2 to G2P[4], even though there was no evidence of infection with this serotype. We developed an enzyme immunoassay to measure sequential levels of anti-NSP2 IgG and IgA, using recombinant (heterotypic) NSP2 derived from SA11 (G3P[2]). Anti-NSP2 IgG and IgA were detected in sera from 23/23 (100%) and 18/24 (75%) of children after primary infection, declined to baseline after 100-150 days, were boosted after rotavirus re-infections, and again declined to baseline 150 days later. Anti-NSP2 IgA was also detected after primary infection, in duodenal juice from 14/16 (87%), and faecal extract from 11/19 (57%) of children. Sequential estimation of anti-NSP2 EIA levels in sera could be a sensitive index of rotavirus infection and re infection. The potential of anti-NSP2 to limit viral replication after re infection deserves further study. PMID- 18428133 TI - Investigation of the environment and of mothers in transmission of rotavirus infections in the neonatal nursery. AB - A distinct feature of neonatal rotavirus infection is the association of unusual strains that appear to be prevalent only in neonatal units and persist for long periods of time. The main aims of this study were to determine if rotavirus can be detected on environmental surfaces in the neonatal nursery and whether the infection occurs in mothers of infected and uninfected neonates. Thirty rotavirus positive neonates and an equal number of negative neonates were enrolled in this study. Stool samples from 15 mothers in each group and environmental swabs collected from the bed and surfaces around neonates were tested for rotavirus using single round and nested PCR for the VP6 gene. Rotavirus could be detected in environmental swabs using single round PCR for VP6 gene in 40% of neonates positive for rotavirus antigen by enzyme immunoassay (EIA) and 33.3% of EIA negative neonates. The detection rate was almost 100% using the nested VP6 PCR. Rotavirus was detected in maternal samples only if the nested VP6 PCR was used, with no significant difference between rates of rotavirus detection in maternal fecal samples of infected and uninfected neonates (p-0.4). Sequence analysis of nested VP6 amplicons from two environmental swabs revealed them to be closest in identity to G10P[11], the most common genotype causing infections in neonates in this setting. Interestingly, sequences of amplicons from maternal stool samples did not cluster with G10P[11] or other VP6 subgroup I strains but showed clustering with human strains of VP6 subgroup II. PMID- 18428134 TI - Lack of protein expression of the Simian virus 40 large T antigen in human lymphomas. AB - Several studies have detected Simian virus 40 (SV40) deoxyribonucleic acid sequences in human tumor tissues, including lymphomas, mainly by the polymerase chain reaction, but these data were not confirmed by subsequent investigations. Regional differences in the distribution of the SV40 and/or technical difficulties have been taken into account to explain these divergent results, but because only a few such studies dealt with the expression of SV40 proteins in tumor tissues, we investigated the expression of the SV40 large T antigen in human lymphomas by immunohistochemistry. Tissue microarrays containing Non Hodgkin's-lymphomas and Hodgkin's-lymphomas were constructed utilizing archival samples encompassing the years 1974--2001 from Italian, Swiss and Austrian patients. Expression of the SV40 large T antigen was analysed by highly specific and sensitive immunohistochemistry using a mouse monoclonal antibody. Protein expression of the large T antigen was not detected in 655 Non-Hodgkin's-lymphomas or in 337 Hodgkin's- lymphomas. The results suggest the absence of an association between SV40 large T antigen and human lymphomas. PMID- 18428135 TI - Genotypes of varicella-zoster virus wild-type strains in Germany. AB - Surveillance of varicella-zoster virus (VZV) genotypes is indicated in Germany after implementation of universal varicella vaccination. This article reports genotyping data of 77 VZV strains obtained from 54 patients with varicella, 1 newborn with congenital varicella syndrome, 2 fetuses with intrauterine VZV infection and 20 cases with zoster. Fragments of the open reading frames (ORF) 1, 21, 22, 37, 50, 54, and 60 were analyzed by sequencing. In addition, the PstI polymorphism of the ORF 38 was characterized. Thirty strains, 22 from varicella and 8 from zoster, had the genetic markers of genotype E2, 2 of them carried new single nucleotide polymorphisms (SNP). Twenty-nine VZV isolates, 17 from varicella, and 12 from zoster, could be analyzed as E1 strains, 6 of them as E1 variants containing individual SNPs. Finally, 17 strains taken from primary VZV infection were classified as genotype M1, 13 of which belonged to the M1 subtype 1, 3 to the M1 subtype 2, and 1 to the M1 subtype 3. One strain was regarded as potential E2/J recombinant. In conclusion, VZV genotypes E2, E1, and M1 can be found in nearly equal incidence in varicella in Germany. The most frequent group is attributed to the genotype E2. Genotype M1 strains can only be detected after primary VZV infection and not in zoster cases. The possible recombinant could not be classified definitely by the scattered SNP method used and, therefore, has to be confirmed by full-genome sequencing studies. PMID- 18428137 TI - Clinical impact of human metapneumovirus genotypes and genotype-specific seroprevalence in Yamagata, Japan. AB - The clinical impact of human metapneumovirus (hMPV) genotypes and the relation between the hMPV genotype in circulation and genotype-specific seroprevalence are yet to be clarified. We determined the genotypes of 93 hMPV strains that were isolated between 2004 and 2006 in Yamagata, Japan, and identified 35 genotype A2, 14 genotype B1, and 44 genotype B2 isolates. Children infected with genotype A2 hMPV were significantly older than those infected with genotype B1 hMPV. Diagnosis of laryngitis was more common in children with genotype B1 hMPV infection and wheezing was more prevalent in children with genotype B1 and B2 hMPV infection than in those with genotype A2 hMPV infection. We then examined genotype-specific seroprevalence by neutralization assay. The higher seropositive rate for the B2 genotype among the children aged 1-2 years is likely to reflect the outbreak of B2 genotype strains in the previous year in this community. The low seropositive rate for the B1 genotype among children aged 1-2 years appears to be associated with a finding that more than 70% of children infected with the B1 genotype were less than 3 years old. In conclusion, we found that the different clinical characteristics of hMPV infection may be associated with hMPV genotype, and the predominant genotype during a season and the affecting age may be closely related to genotype-specific immune status within a community. PMID- 18428138 TI - Unexpected dramatic increase in CD4+ cell count in a patient with AIDS after enfuvirtide treatment despite persistent viremia and resistance mutations. AB - An unexpected dramatic immune recovery was observed in a patient with full-blown AIDS receiving enfuvirtide-based antiretroviral therapy after multiple treatment failures. A complex interplay of viral and host factors, including the control of X4 viruses and proviral burden, may favor immune restoration with HIV neutralizing activity, despite persistent viremia. PMID- 18428140 TI - Changes in circulation of B and non-B HIV strains: spotlight on a reference centre for infectious diseases in Northern Italy. AB - Stored demographic data and HIV RT and protease sequences of 877 HIV patients attending for the first time the HIV/AIDS outpatient clinics of a reference Infectious Diseases centre in Northern Italy between 1999 and 2006 were stratified by 3-year spanning periods according to date of HIV infection. In the period 1980--1982, new infections were entirely caused by HIV-1 subtype B strains and were all diagnosed in injection drug users, 88.9% of whom were males. Injection drug users accounted for 12.8% of new infections in 2004--2006. The frequency of heterosexually-transmitted infections consistently increased until 2000 (from almost none to 51.5%) remaining stable afterwards. About half of heterosexual patients were females. HIV infections among homosexual men increased from 0% in 1980--1982 to 15-21% between 1998 and 2006. Overall, the frequency of non-B subtypes HIV strains increased from 0% in 1980--1982 to 20.3% in 2004--2006 with a greater impact in heterosexuals (from 0% in 1980--1982 to 30.5% in 2004- 2006). In conclusion, a picture of the changing scenario of circulating HIV types and subtypes in a reference Infectious Diseases centre in Northern Italy over the past 26 years is provided. A progressive modification in risk factors for HIV infection and a significant increase in the frequency of non-B HIV strains were observed. PMID- 18428139 TI - Zidovudine with nevirapine for the prevention of HIV mother-to-child transmission reduces nevirapine resistance in mothers from the Western Cape, South Africa. AB - In the Western Cape province of South Africa, an intensified regimen for the prevention-of-mother-to-child-transmission-of-HIV consisting of zidovudine (AZT) from 34 weeks of pregnancy plus single dose (sd) nevirapine (NVP) during labor was instituted in 2004. The newborn baby receives a single dose of NVP and AZT for 7 days. Similar strategies in Thailand and Africa have been shown to be more effective in reducing transmission than NVP alone. The use of sd NVP only for the prevention-of-mother-to-child-transmission-of-HIV has a high risk of inducing resistance (25-69%) with an average of 35.7% by a recent meta-analysis and has been shown to adversely affect non-nucleoside reverse transcriptase inhibitor (NNRTI)-based antiretroviral therapy when initiated within 6 months. In this study the prevalence of resistance to NVP and AZT in mothers who had received the intensified regimen was measured. Specimens collected from mothers were genotyped by in-house PCR and sequencing. In specimens obtained within 60 days of delivery, acquired NVP resistance mutations were detected in 13 of 76 patients (17.1%, 95% confidence interval: 8.7-25.6%), which appears to be lower than in studies with sd NVP alone (37.5%, 95% confidence interval: 23.0-50.6%). PMID- 18428141 TI - High prevalence of hepatitis B virus infection in patients with B-cell non Hodgkin's lymphoma in Korea. AB - This study assessed the association of hepatitis B virus (HBV) and hepatitis C virus (HCV) infection with non-Hodgkin's lymphoma in a highly HBV-endemic area. The prevalence of either HBV or HCV infection in 235 patients with non-Hodgkin's lymphoma was compared with that of an age- and sex-matched hospital control group of 235 patients. The prevalence of HBV infection was higher in B-cell non Hodgkin's lymphoma (15.5%) than control (8.1%), but the prevalence of HCV infection in the non-Hodgkin's lymphoma patients (2.1%) and control group (3%) was similar. HBV prevalence increased significantly with age in the B-cell non Hodgkin's lymphoma patients. The presence of HBV proteins and DNA in lymphoma tissues and peripheral blood mononuclear cells (PBMCs) from HBV-infected non Hodgkin's lymphoma patients was also investigated using immunohistochemistry and PCR. HBV DNA was frequently detected in PBMCs from HBV-infected non-Hodgkin's lymphoma patients, but HBV antigens were not. Therefore, HBV infection, but not HCV infection, was associated with B-cell non-Hodgkin's lymphoma in Korea, suggesting a possible role for HBV in the development of non-Hodgkin's lymphoma. PMID- 18428143 TI - Cervical and oral human papillomavirus types in HIV-1 positive and negative women with cervical disease in South Africa. AB - This study tested cervical and oral human papillomavirus (HPV) infection in HIV-1 seropositive (HIV+) and seronegative (HIV-) women to determine any association between infections at both sites and the difference in prevalence of the HPV types infecting these women. Participants were 115 women referred to a colposcopy clinic after diagnosis of abnormal cervical cytology. The women showed low grade cervical intraepithelial neoplasia (CIN1) or high grade disease (CIN2/3) or no CIN based on colposcopy and histology. Typing of HPV in cervical and oral cells was by Roche linear array and included direct sequencing on selected oral samples. Cervical HPV prevalence was 86.5% and 97.1% in HIV- and HIV+ women respectively. With the exception of HPV-45, prominent in HIV+ women, the hierarchy of predominant types were similar in HIV- and HIV+ women. HPV-16 was most prevalent in both HIV+ (41.7%) and HIV- women (38.5%) with CIN2/3. Significantly more HIV+ women had multiple cervical (>1) infections than HIV- women (36.1% vs. 88.2%, P < 0.001) and more oral HPV infections (45.5% and 25% respectively; P = 0.04). The most prevalent oral HPV types were HPV-33, -11, and 72. The majority of women did not have concordant oral and cervical HPV types, reflecting possible independence of infection at the two sites. HIV immune suppression did not impact significantly on the predominant types of cervical HPV infection (except for HPV-45). HIV+ women had more multiple HPV infections and those with severe cervical disease a similar prevalence of HIV-16 but a lower HPV 18 prevalence than HIV- women. PMID- 18428142 TI - Analysis of the entire nucleotide sequence of hepatitis B causing consecutive cases of fatal fulminant hepatitis in Miyagi Prefecture Japan. AB - We encountered five consecutive patients with fulminant hepatitis induced by acute hepatitis B virus (HBV) infection in 2000--2001 in Japan. They had not had previous contact each other, and were referred to us from different hospitals. Although a 69-year-old woman could be rescued by intensive internal treatment, the four patients died. We analyzed the partial (nt 278-646) and entire nucleotide sequences of the HBV obtained from them, and their divergences were 0 0.3% and 0-0.2%, respectively. The results suggested that they had been infected with the same HBV isolates. The isolates belonged to genotype B and subgenotype B2 on the phylogenetic tree analysis (AB302942-AB302946). As for the nucleotides sequences of them, previously reported mutations of G1896A, A1762T, and G1764A were present. Amino acid analysis revealed that previously reported Ile97Leu and Pro130Non-Pro in the core region and Trp28Stop in the precore region were present. As for the entire nucleotide sequences among B2, AB302942 showed low divergences with AF121245 and AB073834 (1.7%), and X97850 from patients with fulminant hepatitis (3.2%). We compared the two consensus nucleotides derived from AB302942 and X97850 (fulminant hepatitis) versus AY121245 and AB073834 (non fulminant hepatitis), which revealed a difference in nt 1,504 located in the P and X region. Nucleotide 1,504 was C for isolates from fulminant hepatitis and G for non-fulminant hepatitis, and it was recognized among most of the isolates belonging to B2 registered on GenBank. Further studies could disclose the mechanism of severe inflammation of liver that finally leads to fulminant hepatitis. PMID- 18428144 TI - What is the role of serology for the study of chronic hepatitis B virus infection in the age of molecular biology? AB - To assess quantitative serology in chronic hepatitis B virus (HBV) infection, testing by novel immunoassays has been carried out on 202 specimens from untreated patients and in 83 samples from 10 patients with chronic hepatitis B treated with lamivudine. Serum samples were assayed for quantitative HBsAg, in comparison with quantitative HBV-DNA, and for anti-HBc IgM and the avidity index (AI) of total anti-HBc antibodies. The AI was high (mean: 0.93 +/- 0.19) in all groups, confirming the consistency of this procedure in chronic HBV infections. A low-level positivity (2-28 Paul-Ehrlich units/ml) for IgM anti-HBc was detectable both in HBeAg-positive and in HBeAg-negative untreated chronic hepatitis cases (mean S/CO values by the Abbott Architect assay: 0.51 +/- 0.12 and 0.48 +/- 0.10, respectively; correlation between assays: r = 0.685), while treated patients (mean: 0.20 +/- 0.15) and inactive carriers (mean: 0.17 +/- 0.21), were generally negative for IgM. The levels of HBsAg (IU/ml) showed a weak correlation with HBV DNA (IU/ml). A difference in HBsAg levels was found between inactive carriers (1,935 +/- 2,887 IU/ml) and chronic hepatitis B cases, either treated (5,199 +/- 9,259 IU/ml) or untreated (14,596 +/- 15,227 IU/ml). Pre-treatment levels of HBsAg in patients undergoing lamivudine treatment were correlated with a sustained response to therapy over 13-33 months (mean: 27.3) of follow-up: mean HBsAg values were 1,576 + 1,487 IU/ml in five responders and 6,063 + 5,142 in five nonresponders or breakthrough responders (P < 0.05). The availability of standardized quantitative immunoassays for HBsAg and anti-HBc IgM may be considered in addition to quantitative HBV-DNA in the staging and monitoring of chronic HBV infection. PMID- 18428145 TI - Therapy with interferon-alpha and ribavirin for chronic hepatitis C virus infection upregulates membrane HLA-ABC, CD86, and CD28 on peripheral blood mononuclear cells. AB - Multiple interferon-stimulated genes (ISGs) involving T-cell activation are upregulated during initial interferon-alpha-based therapy for chronic hepatitis C virus (HCV) infection. However, the long-term impact on therapeutic outcome in patients remains unknown. In this study, the effects of anti-HCV therapy on the surface expression of HLA-ABC, CD86, and CD28 were longitudinally assessed. These proteins are integral membrane receptors of antigen presentation and triggering of costimulatory signals for activating CD8+ T cells. Peripheral blood mononuclear cells were collected at baseline and post-treatment for 1 day, and 2, 4, 12, and 24 weeks, respectively. This treatment led to a time-related elevation of membrane levels of HLA-ABC and CD86 on B-cells and monocytes in patients with a sustained response (n = 23), but not in those without (n = 8). Meanwhile, upregulation of CD28 on CD4+ and CD8+ T cells was comparable in both groups of sustained responders and non-responders. Steady increases in the B cells' surface and intracellular HLA-ABC were observed, thus, the surface-to-intracellular ratios did not alter over the period of treatment. Furthermore, multivariate analysis shows that increased HLA-ABC on monocytes by week 12 correlates significantly with sustained response (P = 0.033). In conclusion, differential modulation of T-cell activation ISGs, such as HLA-ABC and CD86 might correlate with the outcome of interferon-alpha-based therapy in chronic hepatitis C patients. PMID- 18428146 TI - Predictive factors for cytomegalovirus reactivation in cytomegalovirus seropositive kidney-transplant patients. AB - The aims of the present study were to assess the incidence of cytomegalovirus (CMV) reactivation, and to determine the predictive factors for CMV reactivation in CMV seropositive kidney-transplant patients. One hundred ninety CMV seropositive kidney-transplant patients were included in this study; of these, 39 patients had received CMV prophylaxis. CMV DNAemia was assessed by real timepolymerase chain reaction assay every 2 weeks until day 120, then every 3-4 weeks until day 180, and then every month until day 365. One hundred seven patients (56.3%) had at least one positive CMV DNAemia within the first year. The time between renal transplantation and the first positive CMV DNAemia was 59 +/- 5 days. The number of positive CMV DNAemia/patient was 3.28 +/- 0.22. CMV viral load at first positive CMV DNAemia was 704 (10-742,000) copies/ml. The donor CMV seropositivity, the absence of CMV prophylaxis, and the occurrence of acute rejection before CMV reactivation were independent factors associated with CMV reactivation within the first year after kidney transplantation. Hence, CMV reactivation is frequent after kidney transplantation. CMV prophylaxis in CMV seropositive kidney-transplant patients should be offered to avoid CMV-related side-effects. PMID- 18428147 TI - Development of RNA interference (RNAi) as potential antiviral strategy against enterovirus 70. AB - Enterovirus 70 (EV70) is recognized as the main causative agent of acute hemorrhagic conjunctivitis (AHC), a highly contagious viral infection of the eye. Currently, there is no available treatment for EV70 infections. In this study, we developed a potential intervention strategy using RNA interference (RNAi) against EV70 infection in an in vitro system. Two synthetic 19-mer siRNAs, si-3D1 and si 3D2, were designed to target the 3Dpol region of the EV70 genome. Significant dosage dependent inhibition of EV70 in rhabdomyosarcoma cell line, as shown by reduction of viral RNA and VP1 production, was observed. Both siRNAs prevented EV70 replication in RD cells when transfected into these cells 48 hr prior to virus infection. Introduction of these siRNAs into RD cells 1-3 hr after infection with EV70 reduced production of viral RNA by approximately 60%. Thus, RNAi is a promising strategy to prevent EV70 infections and may have therapeutic potential. PMID- 18428148 TI - Human herpesvirus-6 infection: a prospective study evaluating HHV-6 DNA levels in liver from children with acute liver failure. AB - The aim of this prospective study was to investigate the role of HHV-6 infection in children with acute onset of liver failure using real-time quantitative PCR. Twenty-three children (median age, 24 months) were included: 6 cases of fulminant hepatic failure of undetermined cause (group 1); 4 cases of fulminant hepatic failure of recognized cause (group 2); 3 cases of acute decompensation of chronic liver disease (group 3); and 10 cases of chronic liver disease (group 4). HHV-6 genomic DNA was detected and quantified using real-time PCR in plasma and livers obtained at the time of transplantation. HHV6-DNA detection rate was significantly higher among groups 1, 2, and 3 compared to group 4 (76.9% vs. 20% P = 0.02). Viral loads ranged from 6 to 32,500 copies/106 cells. Significantly higher viral loads were found in 4 of 9 children with acute onset of liver failure of unknown origin (group 1, n = 3; group 3, n = 1) and 1 child with fulminant autoimmune hepatitis (group 2) (P = 0.03). These results strongly support the hypothesis that HHV-6 may cause fulminant hepatic failure and acute decompensation of chronic liver disease in children. Nevertheless, a threshold viral load value still remains to be determined. PMID- 18428149 TI - Impaired cytokine response in myeloid dendritic cells in chronic hepatitis C virus infection regardless of enhanced expression of Toll-like receptors and retinoic acid inducible gene-I. AB - Dendritic cells utilize various sets of Toll-like receptors (TLR) or cytosolic sensors to detect pathogens and evoke immune responses. In patients with hepatitis C virus (HCV) infection, a higher prevalence of various infectious diseases is reported; suggesting that innate immunity against pathogens is impaired. The aim of this study was to clarify whether the TLR and retinoic acid inducible gene-I (RIG-I) system in myeloid dendritic cells is preserved or not in chronic HCV infection. The expression of TLRs, RIG-I and its relatives were compared in myeloid dendritic cells between 39 patients and 52 healthy volunteers. The induction of type-I interferon (IFN) and inflammatory cytokines was examined in response to agonists for TLR2 (palmitoyl-3-cysteine-serine-lysine 4), TLR3/RIG-I (polyinosine-polycytidylic acid) or TLR4 (lipopolysaccharide). The relative expressions of TLR2, TLR4, RIG-I, and LGP2 from the patients were significantly higher than those from the volunteers, whereas TLR3 and MDA-5 expressions did not differ. In search for factors regulating TLR/RIG-I expression, it was shown that IFN-alpha, polyinosine-polycytidylic acid and lipopolysaccharide induced TLR3, TLR4 and RIG-I, but TNF-alpha, HCV core or HCV non-structural proteins did not. For the functional analyses, myeloid dendritic cells from the patients induced significantly less amounts of IFN-beta, TNF-alpha and IL-12p70 in response to polyinosine-polycytidylic acid or lipopolysaccharide. It is noteworthy that the expression of TRIF and TRAF6, which are essential adaptor molecules transmitting TLR3 or TLR4-dependent signals, is reduced in the patients. Thus, innate cytokine responses in myeloid dendritic cells are impaired regardless of enhanced expressions of TLR2, TLR4, and RIG-I in HCV infection. PMID- 18428150 TI - Bruce Merrifield's contribution to antimicrobial peptide research. PMID- 18428151 TI - The early years: working at the bench with Bruce (1957-1959). PMID- 18428152 TI - A tribute to the career of a gentle giant of peptide science. PMID- 18428153 TI - DDAVP (Desmopressin) and solid phase peptide synthesis. PMID- 18428154 TI - Remembering Bruce Merrifield. Collection of reminiscences arranged by Svetlana Mojsov. PMID- 18428155 TI - Festschrift dedicated to the memory of Bruce Merrifield. PMID- 18428156 TI - Bibliography of Bruce Merrifield: reviewed papers, reviews, book chapters, patents, symposium abstracts and articles. PMID- 18428158 TI - Remembering Bruce Merrifield. PMID- 18428157 TI - Working with Bruce Merrifield: the later years (1986-1987). PMID- 18428159 TI - Personal reminiscences of Bruce Merrifield. PMID- 18428160 TI - Editorial: a tribute to Bruce Merrifield. PMID- 18428161 TI - Solid-phase synthesis for a lifetime. PMID- 18428162 TI - Merrifield laboratory: last member standing. PMID- 18428163 TI - The early years: working at the bench with Bruce (1961-1968). PMID- 18428164 TI - Personal reminiscences of Bruce Merrifield. PMID- 18428165 TI - Why do white horses eat more than black horses? PMID- 18428166 TI - Nanoribbons self-assembled from triblock peptide polymers and coordination polymers. PMID- 18428167 TI - Diastereoselective tetrahydropyrone synthesis through transition-metal-free oxidative carbon-hydrogen bond activation. PMID- 18428168 TI - Sensitive NMR sensors detect antibodies to influenza. PMID- 18428169 TI - Photochemical surface patterning by the thiol-ene reaction. PMID- 18428170 TI - Rational engineering of dynamic DNA systems. PMID- 18428171 TI - Au36 crown: a macrocyclization directed by metal-metal bonding interactions. PMID- 18428177 TI - The origin of nonmagnetic Kramers doublets in the ground state of dysprosium triangles: evidence for a toroidal magnetic moment. PMID- 18428172 TI - Control of planar chirality: the construction of a copper-ion-controlled chiral molecular hinge. PMID- 18428178 TI - Synthesis of a polyrotaxane-based macroporous polymer as stationary phase for capillary electrochromatography via host-guest complexation of N,N ' ethylenedianilinediacrylamide with statistically methylated beta-cyclodextrin. AB - A synthetic route to acrylamide-based monolithic stationary phases for CEC with rotaxane-type immobilized derivatized beta-CD was explored. N,N' Ethylenedianilinediacrylamide was synthesized as the water-insoluble crosslinker forming water-soluble inclusion complexes with statistically methylated beta-CD. Mixed-mode stationary phases were synthesized by free radical copolymerization of the bisacrylamide-CD host-guest complex with water-soluble monomers and an additional water-soluble crosslinker in aqueous solution. Complex formation in solution and inclusion of the pseudorotaxane into the polymeric network (formation of a polyrotaxane architecture) were studied by means of (1)H-NMR chemical shift analysis, CD modified micellar EKC (CD-MEKC), 2D-NOESY spectroscopy, and solid state( 13)C-NMR spectroscopy. The presence of a mixed mode selectivity of the stationary phase based on hydrophobic and hydrophilic interaction was confirmed by CEC with neutral polar and nonpolar solutes. PMID- 18428179 TI - HILIC for separation of co-eluted flavonoids under RP-HPLC mode. AB - Hydrophilic interaction chromatography (HILIC) was employed to separate the co eluted flavonoids from licorice extract under RP-HPLC mode. HILIC separations were carried out with the Atalantis HILIC Silica column and the CD-based column. The co-eluted flavonoids were well retained and separated on the two HILIC columns under HILIC mode. Similar results were obtained in the separation of another isoflavones sample, from kudzu extract under HILIC mode. PMID- 18428180 TI - The application of novel 1.7 microm ethylene bridged hybrid particles for hydrophilic interaction chromatography. AB - An un-derivatized 1.7 microm ethylene bridged hybrid (BEH) particle was evaluated for its utility in retaining polar species in hydrophilic interaction chromatography (HILIC), and was compared to a 3 microm un-derivatized silica material. Retentivity as a function of mobile phase pH, polar modifier and ACN content was examined. Also, the efficiency of the two particle substrates was compared by plotting HETP vs. linear velocity. Improved chemical resistance of the un-derivatized BEH particle was compared to un-derivatized silica at pH 5, demonstrating no performance deterioration over the course of 2000 injections for the BEH particle, while the silica particle deteriorated rapidly after 800 injections. Lastly, ESI-MS sensitivity as a function of particle size and separation mode was demonstrated. A 2.2 to 4.7-times higher ESI-MS response was observed on the 1.7 microm particle compared to the 3 microm particle, whereas a 5.6 to 8.8-times higher ESI-MS response was observed using HILIC as when compared to traditional RP chromatography. PMID- 18428181 TI - Stationary phases for hydrophilic interaction chromatography, their characterization and implementation into multidimensional chromatography concepts. AB - Hydrophilic interaction chromatography (HILIC) is becoming increasingly popular for separation of polar samples on polar columns in aqueous-organic mobile phases rich in organic solvents (usually ACN). Silica gel with decreased surface concentration of silanol groups, or with chemically bonded amino-, amido-, cyano , carbamate-, diol-, polyol-, or zwitterionic sulfobetaine ligands are used as the stationary phases for HILIC separations, in addition to the original poly(2 sulphoethyl aspartamide) strong cation-exchange HILIC material. The type of the stationary and the composition of the mobile phase play important roles in the mixed-mode HILIC retention mechanism and can be flexibly tuned to suit specific separation problems. Because of excellent mobile phase compatibility and complementary selectivity to RP chromatography, HILIC is ideally suited for highly orthogonal 2-D LC-LC separations of complex samples containing polar compounds, such as peptides, proteins, oligosaccharides, drugs, metabolites and natural compounds. This review attempts to present an overview of the HILIC separation systems, possibilities for their characterization and emerging HILIC applications in 2-D off-line and on-line LC-LC separations of various samples, in combination with RP and other separation modes. PMID- 18428182 TI - In-line coupling of a reversed-phase column to cope with limited chemoselectivity of a quinine carbamate-based anion-exchange type chiral stationary phase. AB - A chiral stationary phase based on tert-butylcarbamoyl quinine has shown remarkable enantiomer separation capability for the thyroid hormone thyroxine (T(4)) and its structural analogue triiodothyronine (T(3)) employing hydroorganic buffered mobile phases (typical RP conditions). To overcome the problem of a somehow limited chemoselectivity for the critical peak pair between adjacent L thyroxine (L-T(4)) and D-thyroxine (D-T(4)) peaks on the chiral anion-exchanger CSP when all four compounds need to be analysed simultaneously like in impurity profiling of L-T(4 )drug products, an RP column (Gemini C18) was serially coupled with the chiral anion-exchanger column to add a hydrophobic selectivity increment and to improve thereby the critical resolution between L-T(3) and D-T(4). Various parameters such as organic modifier content, pH, buffer concentration and type, type of achiral column as well as sequence of achiral and chiral column have been investigated with individual and tandem columns. With the optimized conditions and use of the tandem column a significantly improved separation, as compared to the chiral anion-exchanger column alone, with a critical resolution as high as 3.7 and an almost equal band spacing of the four components of the test mixture could be obtained. The sequence of the columns (achiral-chiral or chiral-achiral) had no significant effect on the separation performance. PMID- 18428183 TI - Hydrophilic interaction chromatography of small metal species in plants using sulfobetaine- and phosphorylcholine-type zwitterionic stationary phases. AB - Hydrophilic interaction chromatography (HILIC) was applied for the separation of small, noncovalent metal species, and free ligands in plants, using two different zwitterionic stationary phases at pH 7.3. Ligands ranged from amino acids and phytosiderophores to organic acids and synthetic chelators like EDTA. Our results confirmed the suitability of zwitterionic stationary phases for the separation of such ligands and their metal complexes. In particular, the chromatographic behavior of phytosiderophore complexes of copper, nickel, and zinc on a sulfobetaine-type material was compared to that on a phosphorylcholine-type material under otherwise identical conditions. A compression of the usable retention range for phytosiderophore species on the latter phase was found, which can be traced back to the reversed charge orientation of the zwitterionic functionalities. Differences were also found for the integrity of more labile metal chelates during separation on the two columns. In particular, Ni-malate could only be analyzed on the sulfobetaine phase, and Cu-glutathione, Ni aspartate, and Ni-citrate complexes were only stable on the phosphorylcholine stationary phase at pH 7.3. Ni-histidine species were only found after lowering the pH to 4-5. The identification of separated species is possible by online ESI MS in the negative ionization mode. PMID- 18428184 TI - In situ UV Raman spectroscopic studies on the synthesis mechanism of zeolite X. PMID- 18428185 TI - Isoquinolin-1-one inhibitors of the MDM2-p53 interaction. AB - p53 has been at the centre of attention for drug design since the discovery of its growth-suppressive and pro-apoptotic activity. Herein we report the design and characterisation of a new class of isoquinolinone inhibitors of the MDM2-p53 interaction. Our identification of druglike and selective inhibitors of this protein-protein interaction included a straightforward in silico compound selection process, a recently reported NMR spectroscopic approach for studying the MDM2-p53 interaction, and selectivity screening assays using cells with the same genetic background. The selected inhibitors were all able to induce apoptosis and the expression of p53-related genes, but only the isoquinolin-1-one based inhibitors stabilised p53. Our NMR experiments give a persuading explanation for these results, showing that isoquinolin-1-one derivates are able to dissociate the preformed MDM2-p53 complex in vitro, releasing a folded and soluble p53. The joint application of these methods provides a framework for the discovery of protein interaction inhibitors as a promising starting point for further drug design. PMID- 18428186 TI - Regulation of mitochondrial dynamics--characterization of fusion and fission genes in the ascomycete Podospora anserina. AB - The filamentous ascomycete Podospora anserina is a model system for studying aging, a complex process that is regulated by multiple factors. Among these, mitochondria were shown to be of crucial importance. Recently, it was shown that the morphology of these organelles, which is dependent on dynamic fusion and fission processes, has profound effects on P. anserina aging. To further analyze this phenomenon, we characterized molecular components of the machinery regulating the dynamic behavior of mitochondria by utilizing transgenic strains in which fission genes (PaDnm1, PaFis1 and PaMdv1) and a fusion gene (PaFzo1) are overexpressed. While overexpression of PaFis1 has no phenotypic effects in the genetic background of the wild type, it surprisingly promotes mitochondrial fusion and decreases the life span in a mutant overexpressing PaDnm1. Remarkably, when grown on synthetic medium, overexpression of PaDnm1 leads to a decreased life span compared to the wild type. Increased expression of PaMdv1 results in the formation of ring-shaped mitochondria, a morphology of these organelles that has not been previously observed in P. anserina. Transformants with elevated PaFzo1 transcript levels show no altered life span, although the age-dependent fragmentation of mitochondria is impaired. PMID- 18428187 TI - Potential of HILIC-MS in quantitative bioanalysis of drugs and drug metabolites. AB - One fundamental requirement for many lead optimization processes is the need for bioanalytical support within pharmaceutical drug discovery and development. Currently, most bioanalytical methods for pharmaceutical analysis employ HPLC coupled with MS/MS. The combination of HPLC and MS/MS detection frequently offers the complete resolution of the dosed compounds from their metabolites and the endogenous interferences to avoid extra efforts for chemical separation and sample clean-up procedures resulting in higher-throughput assays for a series of new chemical entities (NCEs). Hydrophilic interaction chromatography (HILIC) has been demonstrated to be a powerful technique for the retention of polar analytes offering a difference in selectivity compared to traditional RP chromatography. This review summarizes the HILIC-MS/MS methods for the trace quantitative determinations of the drug compounds and their metabolites to support both in vitro and in vivo experiments. The challenges on performing HILIC-MS/MS assays such as matrix ionization suppression and the potential for endogenous interferences are also presented. PMID- 18428188 TI - Evaluation of hydrophilic interaction chromatography versus reversed-phase chromatography in a plant metabolomics perspective. AB - The metabolomics goal, the unbiased relative quantification of all metabolites in a biological system, still lacks a universal analytical approach. In the LC-MS line of approach, one of the major problems encountered is the polar nature of a large group of (plant) metabolites. Here, we investigate the potential of hydrophilic interaction chromatography (HILIC) and compare its qualities with extended polarity RP chromatography. Two opposite LC phase compositions (Atlantis dC18 vs. TSKgel Amide-80) are compared in a plant metabolomics setting. Both performed equally well with regard to retentive capacities, but variation in peak area was about 5% higher for the HILIC approach. Focussing on matrix effects (ME) on the other hand, it was observed that this well-known problem in RP LC-MS metabolomics was not reduced on using hydrophilic interaction chromatography. PMID- 18428189 TI - Hydrophilic interaction liquid chromatography method for the determination of ascorbic acid. AB - Hydrophilic interaction liquid chromatography (HILIC) method using internal standard for the determination and stability study of ascorbic acid was developed. HILIC method was very fast and simple using the following analytical conditions: ZIC HILIC (150 x 2.1 mm, 3.5 microm) chromatographic column and mobile phase composed of ACN and 50 mM ammonium acetate buffer pH 6.8 (78:22 v/v). Diode array detection was performed and chromatograms were processed at 268 nm, the maximum wavelength of absorbance of ascorbic acid. An extensive stability study of ascorbic acid as a function of various factors including temperature, stabilizing agents, oxygen presence and its concentration in solution was performed in order to gain information about the quantitative influence of individual stability factors. Low temperature and stabilizing agents (o phosphoric acid and oxalic acid) were found to be key factors enabling substantial enhancement of the stability of ascorbic acid. PMID- 18428190 TI - Deconvolution of electrokinetic and chromatographic contributions to solute migration in stereoselective ion-exchange capillary electrochromatography on monolithic silica capillary columns. AB - A monolithic silica stationary phase functionalized with an enantioselective strong cation exchanger based on an aminosulfonic acid derivative was used for chiral separations of basic test solutes by nonaqueous CEC and capillary LC. The effects of the applied electric field as well as the ionic strength in the eluent on electrokinetic and chromatographic contributions to the overall separation performance in the electrically driven mode were investigated. Hence, under the utilized experimental conditions, i. e., at an electric field strength in the range of approximately 120-720 V/cm (applied voltages 4-24 kV) and an ionic strength of the counterion between 5 and 25 mM (at constant acid-to-base, i. e., co- to counterion ratio of 2:1), no deviations from the expected linearity of the EOF were observed. This led to the conclusion that an occurrence of the so-called electrokinetic effects of the second kind resulting from electric double layer overlap inside the mesopores of the monolithic stationary phase and concentration polarization phenomena were largely negligible. Additional support to this conclusion was inferred from the observed independence of CEC retention factors on the electric field strength across the investigated ionic strength range of the BGE. As a consequence, a simple framework allowing for calculation of the CEC mobilities from the individual separation contributions, viz. electroosmotic and electrophoretic mobilities as well as retention factors, could be applied to model CEC migration. There was a reasonable agreement between calculated and experimental CEC mobility data with deviations typically below 5%. The deconvolution of the individual contributions to CEC migration and separation is of particular value for the understanding of the separation processes in which electrophoretic migration of ionic sample constituents plays a significant role like in ion-exchange CEC and may aid the optimization procedure of the BGE and other experimental conditions such as the optimization of the surface chemistry of the stationary phase. In combination with the remarkable column performance evident from the low theoretical plate heights observed under CEC conditions for all test solutes (3.5-7.5 microm in the flow rate range of 0.4-1.2 mm/s, corresponding to (130,000-300,000 plates per meter), the presented framework provides an attractive tool as the basis for the assessment of chromatographic selectivities in a miniaturized CEC screening of new selectors and chiral stationary phases (CSPs), respectively, from experimental CEC data and known CE mobilities. PMID- 18428191 TI - Development of retention prediction models for adrenoreceptor agonists and antagonists on a polyvinyl alcohol-bonded stationary phase in hydrophilic interaction chromatography. AB - Retention prediction models based on multiple linear regression (MLR) and artificial neural network (ANN) for adrenoreceptor agonists and antagonists chromatographed on a polyvinyl alcohol-bonded stationary phase under hydrophilic interaction chromatography were described. The models showed the combined effects of solute structure and mobile phase composition on the retention behavior of the analytes. Using stepwise MLR, the retentions of the studied compounds were satisfactorily described by a five-predictor model; the predictors being the %ACN, the logarithm of the partition coefficient (log D), the number of hydrogen bond donors (HBD), the desolvation energy for octanol (FOct), and the total absolute atomic charge (TAAC). The inclusion of the solute-related descriptors suggested that hydrophilic interactions such as hydrogen bonding and also ionic interactions are possible mechanisms by which analytes are retained on the studied system. ANN prediction models were also derived using the predictors derived from MLR as inputs and log k as outputs. The best network architectures were found to be 5-3-1 for the datasets at pH 3.0 and 4.0, and 5-4-1 for the dataset at pH 5.0. The optimized ANNs showed better predictive properties than the MLR models for both training and test sets under all pH conditions. PMID- 18428193 TI - Potential role of mind-body therapies in cancer survivorship. AB - The use of complementary and alternative medicine (CAM) by cancer survivors is high, particularly among those with psychosocial distress, poor quality of life, culturally based health beliefs, and those who experience health disparities in the mainstream healthcare system. As the number of cancer survivors continues to increase, so does the diversity of the survivorship population, making it increasingly important to understand and address the CAM culture in different survivor groups. Given the known communication barriers between cancer patients and their physicians regarding CAM, it would be useful for oncology providers to have a platform from which to discuss CAM-related issues. It is proposed that mind-body therapies with some basis in evidence could provide such a platform and also serve as a possible means of connecting cancer survivors to psychosocial supportive services. This article reviews a few mind-body therapies that may have particular relevance to cancer survivors, such as hypnosis and meditation practices. A theoretical foundation by which such therapies provide benefit is presented, with particular emphasis on self-regulation. PMID- 18428192 TI - Recruiting and retaining breast cancer survivors into a randomized controlled exercise trial: the Yale Exercise and Survivorship Study. AB - BACKGROUND: Given observational findings that physical activity reduces breast cancer risk, improves survival, and improves quality of life in breast cancer survivors, a need has been identified for randomized controlled trials that test the efficacy of exercise on biological mechanisms associated with breast cancer survival. The primary aims of the Yale Exercise and Survivorship Study were to 1) determine the feasibility of recruiting breast cancer survivors into a randomized controlled trial of the effects of exercise on biological markers and/or mechanisms associated with survival, 2) compare the effectiveness of various recruitment strategies on accrual rates and baseline characteristics, and 3) report adherence to the exercise trial. METHODS: Seventy-five postmenopausal breast cancer survivors self-referred into the trial or were recruited through the New Haven Tumor [corrected] Registry and randomly assigned to an exercise (n = 37) or usual-care (n = 38) group. The exercise group participated in 150 min/wk of supervised gym-based and home-based aerobic exercise for 6 months. The usual care group was instructed to maintain current physical activity level. RESULTS: A total of 75 women (an accrual rate of 9.5%) were randomized to the trial. Rates of accrual were higher for women who self-referred into the study (19.8%) compared with women recruited via the cancer registry (7.6%); however, demographic, physiologic, and prognostic characteristics did not differ between the 2 recruitment strategies. On average, exercisers increased moderate- intensity to vigorous-intensity aerobic exercise by 129 minutes per week compared with 44 minutes per week among usual-care participants (P < .001). Women in the exercise-intervention group increased their average pedometer steps by 1621 steps per day compared with a decrease of 60 steps per day among women in the usual care group (P < .01). CONCLUSIONS: Findings from this study will provide useful information for investigators who are conducting exercise trials in cancer populations, clinicians who are treating women diagnosed with breast cancer, and exercise professionals who are developing community-based exercise programs for cancer survivors. PMID- 18428194 TI - Survivorship research based in integrated healthcare delivery systems: the Cancer Research Network. AB - BACKGROUND: Integrated healthcare delivery systems present unique opportunities for cancer survivorship research. The National Cancer Institute funds the Cancer Research Network (CRN) to leverage these capabilities for all types of cancer research, including survivorship. METHODS: The authors gathered information from a recent CRN funding application, Survivorship Interest Group materials, the CRN website, and published articles. CRN studies were selected to illustrate diverse topics and a variety of data-collection approaches. RESULTS: The 14 systems that participate in the CRN provide care for approximately 10.8 million individuals of all ages and racial/ethnic backgrounds, for whom approximately 38,000 new cancer diagnoses were made in 2005. CRN systems have the ability to use existing data and collect new data on patients, providers, and organizations through well established research centers staffed by independent scientists. Of the 45 funded and 2 pending CRN grant applications as of November 30, 2007, 21 include aspects related to cancer survivorship. These studies have examined clinical trial participation, patterns of care, age and racial/ethnic disparities, diffusion of clinical trial findings, treatment outcomes, surveillance, and end-of-life and palliative care. Breast, colorectal, lung, ovarian, and prostate cancers have been the focus of these studies. Results of these studies have been published widely in leading journals. CONCLUSIONS: Completed and ongoing CRN survivorship studies provide a strong foundation for future studies. Scientists from all institutional affiliations are welcome to approach the CRN with ideas and are encouraged to allow ample time to establish collaborative relationships and design rigorous studies. PMID- 18428195 TI - Physical activity and obesity in Canadian cancer survivors: population-based estimates from the 2005 Canadian Community Health Survey. AB - BACKGROUND: Physical inactivity and obesity are associated with poorer disease outcomes in several cancer survivor groups. Few studies, however, have provided population-based estimates of these risk factors in cancer survivors and compared them with individuals without a history of cancer. Here such estimates for the Canadian population are reported. METHODS: Data were obtained from the 2005 Canadian Community Health Survey consisting of computer-assisted interviews of 114,355 adults representing an estimated 23,285,548 Canadians. Participants self reported their cancer history, height, and body weight to calculate body mass index and participation in various leisure-time activities. RESULTS: Fewer than 22% of Canadian cancer survivors were physically active and over 18% were obese. Few differences were observed between cancer survivors and those without a history of cancer except that: 1) prostate cancer survivors were more likely to be active (adjusted odds ratio [OR] = 1.27; 95% confidence interval [CI] = 1.01 1.59) and less likely to be obese (adjusted OR = 0.71; 95% CI = 0.56-0.90); 2) skin cancer survivors (nonmelanoma and melanoma) were more likely to be active (adjusted OR = 1.33; 95% CI = 1.12-1.59); and 3) obese breast cancer survivors were less likely to be active compared with obese women without a history of cancer (adjusted OR = 0.51; 95% CI = 0.27-0.94). CONCLUSIONS: Canadian cancer survivors have low levels of physical activity and a high prevalence of obesity that, although comparable to the general population, may place them at higher risk for poorer disease outcomes. Population-based interventions to increase physical activity and promote a healthy body weight in Canadian cancer survivors are warranted. PMID- 18428196 TI - Limits of observational data in determining outcomes from cancer therapy. AB - BACKGROUND: Observational data are used increasingly to assess the effectiveness of therapies. However, selection biases are likely to have an impact on results and threaten the validity of these studies. METHODS: The primary objective of the current study was to explore the effect of selection biases in observational studies of treatment effectiveness in cancer care. Patients were identified from the Surveillance, Epidemiology, and End Results-Medicare linked database. The following groups of patients were included: 5245 men treated with and without androgen deprivation for locally advanced prostate cancer, 43,847 men with active treatment versus observation for low- and intermediate-risk prostate cancer, and 4860 patients with lymph node-positive colon cancer who were treated with and without fluorouracil chemotherapy. Patients were compared by therapy for the outcomes of cancer-specific mortality, other-cause mortality, and overall mortality. RESULTS: In all comparisons, the observational data produced improbable results. For example, when evaluating outcomes of men who were treated with and without androgen deprivation for locally advanced prostate cancer, men who underwent androgen deprivation had higher prostate cancer mortality (hazard ratio, 1.5; 95% confidence interval, 1.29-1.92) despite clinical trial evidence that this treatment improves cancer mortality. Controlling for comorbidity, extent of disease, and other characteristics by multivariate analyses or by propensity analyses had remarkably small impact on these improbable results. CONCLUSIONS: The current results suggested that the results from observational studies of treatment outcomes should be viewed with caution. PMID- 18428197 TI - Development of a high drug load tablet formulation based on assessment of powder manufacturability: moving towards quality by design. AB - The development of a robust tablet formulation for a high dose active pharmaceutical ingredient (API) by the trial-and-error approach is challenging. To meet the growing needs of bringing drugs to market faster and with reduced costs, more targeted and efficient development practices are in demand. Here we show detailed understanding of mechanical properties of API and excipients are essential in achieving efficient development of a high API loading formulation. The loading of the experimental drug, AMG458, was 50 wt% plus accompanying 1:1 molar ratio organic acid of approximately 19%. We assessed manufacturability of powders based on their flow and compaction properties using a shear cell and a compaction simulator, respectively. We selected granulation process on the basis of poor flow properties of API and its blends with common direct compaction excipients. During the course of formulation development, we could quickly identify manufacturability deficiencies in the lead formulation. With detailed knowledge of the mechanical properties of excipients and formulated powders, we improved the lead formulation by overcoming manufacturability deficiencies using predictive and material sparing (<10 g) approaches. Larger batches were subsequently manufactured to confirm predictions. PMID- 18428198 TI - C-reactive protein as a prognostic marker for men with androgen-independent prostate cancer: results from the ASCENT trial. AB - BACKGROUND: Studies of cancer risk and molecular carcinogenesis suggest a role for inflammation in cancer development and progression. The authors sought to determine whether specific blood proteins associated with inflammation predict for outcomes in men with metastatic androgen-independent prostate cancer (AIPC) who are initiating docetaxel-based chemotherapy. METHODS: Baseline plasma samples were stored (-80 degrees C) from 160 of 250 patients enrolled in the AIPC Study of Calcitriol ENhancing Taxotere (ASCENT) trial, a randomized, placebo-controlled trial comparing weekly docetaxel plus high-dose calcitriol with weekly docetaxel. Multiplex immunoassays measured 16 cytokine, chemokine, cardiovascular, or inflammatory markers. The Cox proportional hazards model was used to assess associations between baseline biomarkers, clinical characteristics, and survival. Logistic regression was used for analyses of associations with prostate-specific antigen (PSA) decline. RESULTS: C-reactive protein (CRP) was found to be significantly predictive of a shorter overall survival (hazards ratio [HR] of 1.41 for each natural logarithm [ln] [CRP] increase; 95% confidence interval [95% CI], 1.20-1.65 [P < .0001]). When CRP (continuous) was entered into a multivariate model using 13 baseline clinical variables, only elevated CRP remained a significant predictor (P < .0001) of shorter overall survival. When categorized as normal (8 mg/L), elevated CRP was found to be a significant predictor of shorter overall survival (HR of 2.96; 95% CI, 1.52-5.77 [P = .001]), as was hemoglobin (P = .007). Elevated CRP was also associated with a lower probability of PSA decline (odds ratio of 0.74 for each ln(CRP) increase; 95% CI, 0.60-0.92 [P = .007]). CONCLUSIONS.: Elevated plasma CRP concentrations appear to be a strong predictor of poor survival and lower probability of PSA response to treatment in patients with AIPC who are receiving docetaxel-based therapy. PMID- 18428199 TI - Quality of life of family caregivers of cancer survivors: across the trajectory of the illness. AB - BACKGROUND: Cancer affects not only the quality of life (QOL) of individuals with the disease but also that of their family members and close friends. The impact on various aspects of the family caregivers' QOL is significant throughout the trajectory of the illness. The authors reviewed literature on the QOL of family caregivers at the acute and middle- to long-term survivorship phases as well as the bereavement phase. METHODS: By using several databases, the authors searched the literature published in English from 1996 through July 2007. Keywords searched included cancer, carcinoma, family, family member, caregivers, and quality of life. Several criteria were used to guide the literature review: Articles had to be published in refereed journals and had to use rigorous methods, sample, and validated measures. RESULTS: The findings suggested that the QOL of family caregivers of individuals with cancer varies along the illness trajectory. This highlights were importance of assessing the ongoing adjustment of the caregivers over time. However, there were few theory-driven studies, and significant gaps remain in the current understanding of the effects of family caregiving beyond the time of diagnosis and treatment. CONCLUSIONS: Accumulating evidence has supported the concept that cancer affects not only the patients/survivors but also their family members. However, theoretically and methodologically rigorous research on various aspects of the family's QOL, including not only the psychological but also the physical, spiritual, and behavioral adjustment to cancer in the family, remains sparse. Family-based interventions across the trajectory of the illness also are needed. PMID- 18428200 TI - Introduction: Partnering to embrace the future of cancer survivorship research and care. PMID- 18428201 TI - Information support for cancer survivors. AB - Survivors' information-seeking behavior has traditionally been documented through analysis of inquiries to hotlines and cancer information services. Data from these self-selected inquiries tend to be restricted to a time around diagnosis, and to those populations possessing the wherewithal and motivation to seek information actively. The current study used data from a general population survey to assess 1) the prevalence of information-seeking behavior among survivors in the general population, 2) characteristics of seekers versus nonseekers, 3) ratings of information-seeking experience, and 4) actual versus preferred sources of information. Data were analyzed from the 2005 administration of the Health Information National Trends Survey (HINTS). HINTS is a cross sectional, random digit dial telephone survey, weighted to provide estimates for the general population. Nearly half of all Americans (48.7%) indicated that they have looked for cancer information from 1 source or another. Percentages were highest for those who have been touched by cancer (63.1% of cancer survivors and 54.6% of those with family histories) and lowest for those with no cancer history (27.6% of those with no history). Quality concerns topped the list of information seeking experiences for those recalling the last time they looked. Patterns of information-seeking revealed a discrepancy between preferred and actual source when tracked over years since diagnosis. Information-seeking is prevalent among cancer survivors and does not diminish over time. Prescriptions are given for reengineering the information environment to improve long-term outcomes for survivors. PMID- 18428202 TI - Intimacy and relationship processes in couples' psychosocial adaptation to cancer. AB - The authors highlighted the importance of viewing cancer from a relationship perspective. This perspective not only considers the marital relationship as a resource that individual partners draw upon but also highlights the importance of focusing attention onto the relationship and engaging in communication behaviors aimed at sustaining and/or enhancing the relationship during stressful times. On the basis of existing conceptualizations, empiric research on couples and cancer, and the authors' perspective on the literature, they formulated the relationship intimacy model of couples' psychosocial adaptation to cancer as a first step toward building a framework for researchers and clinicians to inform their work in this area. The model proposes that patients and their partners engage in behaviors that either promote or undermine the level of closeness in their relationship and that the closeness of the marital relationship is an important determinant of patient and partner psychologic adaptation to cancer. Preliminary data from a couples' intimacy-enhancing intervention for breast cancer patients and their partners supported the model. Of the 25 couples who consented to participate in the intervention and completed the preintervention surveys, 15 couples completed all 5 sessions, and 12 couples completed the follow-up survey. The current results suggested that the intervention improved patient and partner perceptions of the closeness of their relationship and reduced their distress. The authors also discuss limitations of the relationship intimacy model as well as future directions for empiric and clinical research on couples' psychosocial adaptation to cancer. PMID- 18428203 TI - Clinical, cellular, and neuropathological consequences of AP1S2 mutations: further delineation of a recognizable X-linked mental retardation syndrome. AB - Mutations in the AP1S2 gene, encoding the sigma1B subunit of the clathrin associated adaptor protein complex (AP)-1, have been recently identified in five X-linked mental retardation (XLMR) families, including the original family with Fried syndrome. Studying four patients in two unrelated families in which AP1S2 nonsense and splice-site mutations segregated, we found that affected individuals presented, in addition to previously described features, with elevated protein levels in cerebrospinal fluid (CSF). Moreover, computed tomography scans demonstrated that the basal ganglia calcifications associated with AP1S2 mutations appeared during childhood and might be progressive. Based on these observations, we propose that AP1S2 mutations are responsible for a clinically recognizable XLMR and autism syndrome associating hypotonia, delayed walking, speech delay, aggressive behavior, brain calcifications, and elevated CSF protein levels. Using the AP-2 complex, in which the sigma subunit is encoded by one single gene, as a model system, we demonstrated that sigma subunits are essential for the stability of human AP complexes. By contrast, no major alteration of the stability, subcellular localization, and function of the AP-1 complex was observed in fibroblasts derived from a patient carrying an AP1S2 mutation. Similarly, neither macro- nor microscopic defects were observed in the brain of an affected fetus. Altogether, these data suggest that the absence of an AP-1 defect in peripheral tissues is due to functional redundancy among AP-1 sigma subunits (sigma1A, sigma1B, and sigma1C) and that the phenotype observed in our patients results from a subtle and brain-specific defect of the AP-1-dependent intracellular protein traffic. PMID- 18428204 TI - A gerontologic perspective on cancer and aging. AB - Most people diagnosed with cancer are aged >65 years, and many diagnosed younger live to become older survivors. Geriatric oncology is becoming recognized as a specialty area within oncology. It focuses specifically on the functional impacts of the interplay of aging and cancer, including the role of comorbidities. Nevertheless, to the authors' knowledge, little attention has been given to cancer from a gerontologic and lifespan perspective, especially quality of life and psychologic impact. Research has shown that the amount and type of psychologic impact of cancer is highly variable and that part of that variation is related to age, in that older persons are often less affected in both negative and positive ways. Gerontologic concepts and empiric findings related to physical, psychologic, and social aging processes may serve as partial explanations for that age-related pattern. Important potential contributors include psychologic factors, such as changes in future time perspective and goals, as well as social ones, such as roles and previous experience. The result is a complex interplay of factors that vary across persons but are covaried with age. Empiric findings regarding 1-year to 8-year prostate cancer survivors illustrate the age differences and the differential impacts of age itself and comorbidity. The use of gerontologic concepts to explain the age-related impact of cancer will benefit both research and clinical practice by providing a means to target interventions more effectively by taking into account the psychologic and social changes that often accompany aging. . PMID- 18428205 TI - Physical and psychological long-term and late effects of cancer. AB - The number of long-term cancer survivors (> or =5 years after diagnosis) in the U.S. continues to rise, with more than 10 million Americans now living with a history of cancer. Along with such growth has come increasing attention to the continued health problems and needs of this population. Many cancer survivors return to normal functioning after the completion of treatment and are able to live relatively symptom-free lives. However, cancer and its treatment can also result in a wide range of physical and psychological problems that do not recede with time. Some of these problems emerge during or after cancer treatment and persist in a chronic, long-term manner. Other problems may not appear until months or even years later. Regardless of when they present, long-term and late effects of cancer can have a negative effect on cancer survivors' quality of life. This article describes the physical and psychological long-term and late effects among adult survivors of pediatric and adult cancers. The focus is on the prevalence and correlates of long-term and late effects as well as the associated deficits in physical and emotional functioning. In addition, the emergence of public health initiatives and large-scale research activities that address the issues of long-term cancer survivorship are discussed. Although additional research is needed to fully understand and document the long-term and late effects of cancer, important lessons can be learned from existing knowledge. Increased awareness of these issues is a key component in the development of follow-up care plans that may allow for adequate surveillance, prevention, and the management of long-term and late effects of cancer. PMID- 18428206 TI - Oral absorption enhancement of dipeptide L-Glu-L-Trp-OH by lipid and glycosyl conjugation. AB - In recent years, the conjugation of sugar moieties and lipoamino acids has been extensively investigated as a mean to enhance the stability towards enzymatic degradation and the permeability across biological membranes of poorly orally available drugs, including peptides. In this prospect, a library of novel derivatives of the dipeptide L-Glu-L-Trp, a naturally occurring thymic immunomodulator with high hydrophilic character and low membrane permeability, was designed and synthesised by conjugating 2-amino-dodecanoic acid (C(12)) and/or 1-amino-beta-D-glucuronic acid (GlcAN), beta-D-glucuronic acid (GlcA) and N-beta-D-glucopyranosylamine succinamic acid (GlsNS) residues to the Glu-Trp scaffold, using an Fmoc solid-phase peptide synthesis strategy on trichlorotrityl resin. A cellobiose derivative was also prepared in solution. The synthesized peptides showed no sign of toxicity to red blood cells at 200 microM (haemolysis assay) and their resistance against enzymatic hydrolysis, assessed in Caco-2 homogenates, was usually significantly increased, particularly for the C-terminal conjugates. Several derivatives also saw their apparent permeability values greatly enhanced and one of the conjugates tested proved to be able to release the initial dipeptide after penetrating Caco-2 monolayers. An initial in vivo experiment was then carried out in male Wistar rats to examine the effect of conjugation on the absorption rate and bioavailability. PMID- 18428207 TI - Structural insights for designed alanine-rich helices: comparing NMR helicity measures and conformational ensembles from molecular dynamics simulation. AB - The temperature dependence of helical propensities for the peptides Ac-ZGG (KAAAA)(3)X-NH(2) (Z = Y or G, X = A, K, and D-Arg) were studied both experimentally and by MD simulations. Good agreement is observed in both the absolute helical propensities as well as relative helical content along the sequence; the global minimum on the calculated free energy landscape corresponds to a single alpha-helical conformation running from K4 to A18 with some terminal fraying, particularly at the C-terminus. Energy component analysis shows that the single helix state has favorable intramolecular electrostatic energy due to hydrogen bonds, and that less-favorable two-helix globular states have favorable solvation energy. The central lysine residues do not appear to increase helicity; however, both experimental and simulation studies show increasing helicity in the series X = Ala --> Lys --> D-Arg. This C-capping preference was also experimentally confirmed in Ac-(KAAAA)(3)X-GY-NH(2) and (KAAAA)(3)X-GY-NH(2) sequences. The roles of the C-capping groups, and of lysines throughout the sequence, in the MD-derived ensembles are analyzed in detail. PMID- 18428208 TI - A novel approach for calculating starch crystallinity and its correlation with double helix content: a combined XRD and NMR study. AB - A peak fitting procedure has been implemented for calculating crystallinity in granular starches. This methodology, widely used for synthetic polymers, is proposed to better reflect the crystalline content of starches than the method normally used, in which it is assumed that relatively perfect crystalline domains are interspersed with amorphous regions. The new approach takes into account irregularities in crystals that are expected to exist in semicrystalline materials. Therefore, instead of assuming that the amorphous background extends up to the base of diffraction peaks, the whole X-ray diffraction (XRD) profile is fitted to an amorphous halo and several discrete crystalline diffraction peaks. The crystallinity values obtained from the XRD patterns of a wide range of native starches using this fitting technique are very similar to the double helix contents as measured by (13)C solid state NMR, suggesting that double helices in granular starches are present within irregular crystals. This contrasts with previous descriptions of crystalline and noncrystalline double helices that were based on the analysis of XRD profiles as perfect crystals interspersed in a noncrystalline background. Furthermore, with this fitting methodology it is possible to calculate the contribution from the different crystal polymorphs of starch to the total crystallinity. PMID- 18428209 TI - Malignant primary cardiac tumors: review of a single institution experience. AB - BACKGROUND: Primary cardiac sarcomas are uncommon. The authors undertook to review the Mayo Clinic's experience with primary cardiac sarcomas consisting of 34 patients seen over a 32-year period. METHODS: The patient database at the Mayo Clinic was searched to identify patients with malignant tumors of the heart seen during the 32-year period between 1975 and 2007. Thirty-four patients with primary cardiac sarcomas were identified and their medical records reviewed for details pertaining to presenting symptoms, staging modalities, treatment approaches, and outcomes. RESULTS: Of the 34 patients, 17 were men and 17 were women. The median age was 44 years. The mean duration of symptoms at the time of diagnosis was 3.6 months. The most common histologic type was angiosarcoma (41%). The median follow-up for the entire group was 12 months (range, 0-61 months). The median survival for those who underwent a complete surgical excision was 17 months compared with 6 months for those in whom a surgical complete remission could not be achieved (P = .01). Patients with angiosarcoma had a lower survival compared with patients with other histologies (5 months vs 17 months; P = .01). The median survival of patients with metastatic disease was 5 months versus 15 months in patients without metastatic disease (P = .03 by the log-rank test). CONCLUSIONS: Cardiac sarcomas remain a rare but lethal disease. Compared with extracardiac sarcomas, the prognosis for patients with cardiac sarcomas remains very poor. A complete surgical excision should be performed if possible. Innovative treatment strategies are required. PMID- 18428210 TI - Prognostic factors for patients with stage IV epithelial ovarian cancer receiving intraperitoneal chemotherapy after second-look assessment: results of long-term follow-up. AB - BACKGROUND: The aim was to determine the long-term outcome for patients with FIGO stage IV epithelial ovarian carcinoma (EOC) treated with intraperitoneal (IP) chemotherapy after second-look assessment. METHODS: By using data from a retrospective cohort of 433 patients who received IP therapy after second-look assessment after primary surgery and initial systemic therapy for EOC between 1984 and 1998 at our institution, all FIGO stage IIIC and IV patients were identified. Standard statistical methods were used. RESULTS: Overall, 297 patients met study criteria (246 stage IIIC; 51 stage IV). The median survival for patients with stage IV disease was 34 months compared with 42 months for patients with stage IIIC disease (P=.02). The only significant predictor of overall survival in patients with stage IV disease was the presence of gross residual disease at initiation of IP therapy (P=.027). When comparing stage IV patients with and without pleural effusions to all stage IIIC patients, there was a significant trend toward improved survival in the patients with pleural effusions only compared with other stage IV patients (P=.01). CONCLUSIONS: Prolonged overall survival was observed in patients with no gross residual disease at the time of IP chemotherapy initiation. When compared with similarly treated stage IIIC patients, stage IV patients with malignant pleural effusions appear to have a better outcome than those with other sites of metastasis. Future prospective trials should evaluate the use of IP therapy for patients with stage IV EOC by virtue of malignant pleural effusions only who responded to initial systemic therapy. PMID- 18428211 TI - Efficacy of lamotrigine in the management of chemotherapy-induced peripheral neuropathy: a phase 3 randomized, double-blind, placebo-controlled trial, N01C3. AB - BACKGROUND: Lamotrigine, an antiepileptic agent, has been reported as being effective in reducing symptoms of neuropathy associated with various etiologies. Based on such data, a multicenter double-blind, placebo-controlled, randomized trial was conducted to evaluate the effect of lamotrigine on pain and other neuropathic symptoms due to chemotherapy-induced peripheral neuropathy (CIPN). METHODS: Patients with symptomatic CIPN with symptom scores of either 1) >3 on a 0-10 Numerical Rating Scale (NRS) or 2) >1 on the 0-3 the Eastern Cooperative Oncology Group (ECOG) neuropathy scale (ENS) were eligible (higher numbers corresponding to greater severity of symptoms in both scales). Patients were randomly assigned to receive lamotrigine (target dose of 300 mg/day) or placebo for 10 weeks. Endpoints were measured biweekly. RESULTS: In all, 131 patients were enrolled. Both groups were well matched at baseline. Over the 10-week period of the trial, the average pain scores (NRS) for the lamotrigine and placebo arms declined in both arms, with no statistically significant difference noted between the changes in the 2 groups (0.3 and 0.5 unit reduction from baseline, respectively; P=.56). Similarly, decreases in the ENS with therapy were not statistically different (0.4 and 0.3, respectively; P=.3). Changes in other subjective symptom scales were also not found to be statistically different between the 2 groups. Toxicities were mild and similar in each group. CONCLUSIONS: The results suggest that lamotrigine is not effective for relieving neuropathic symptoms in patients because of CIPN. PMID- 18428212 TI - Preoperative assessment enables the early diagnosis and successful treatment of lymphedema. AB - BACKGROUND: The incidence of breast cancer (BC)-related lymphedema (LE) ranges from 7% to 47%. Successful management of LE relies on early diagnosis using sensitive measurement techniques. In the current study, the authors demonstrated the effectiveness of a surveillance program that included preoperative limb volume measurement and interval postoperative follow-up to detect and treat subclinical LE. METHODS: LE was identified in 43 of 196 women who participated in a prospective BC morbidity trial. Limb volume was measured preoperatively and at 3-month intervals after surgery. If an increase>3% in upper limb (UL) volume developed compared with the preoperative volume, then a diagnosis of LE was made, and a compression garment intervention was prescribed for 4 weeks. Upon reduction of LE, garment wear was continued only during strenuous activity, with symptoms of heaviness, or with visible swelling. Women returned to the 3-month interval surveillance pathway. Statistical analysis was a repeated-measures analysis of variance by time and limb (P10 to 30 cells. A basic protocol for array hybridization on nylon filters is provided because such filters are available in every laboratory. Tissue samples contain many different cell types in variable amounts, so their analysis may require microdissection; a protocol for obtaining cryosections is given. Finally, a simple procedure to prepare the data for statistical analysis is also provided. PMID- 18428373 TI - An overview of Spotfire for gene-expression studies. AB - Spotfire DecisionSite for Functional Genomics (referred to here as Spotfire) is a powerful data mining and visualization application with use in many disciplines. This unit provides an overview of Spotfire's utility in analyzing gene expression data obtained from DNA microarray experiments. Analysis of microarray data requires software-based solutions able to handle and manipulate the enormous amount of data generated. Spotfire provides a solution for accessing, analyzing and visualizing data generated from microarray experiments. Spotfire is designed to allow biologists with little or no programming or statistical skills to transform, process, and analyze microarray data. PMID- 18428374 TI - Helper-dependent adenoviral vectors. AB - The helper-dependent adenovirus (HDAd) is a recently developed adenovirus-based vector with an improved safety profile and long-term transgene expression. In this unit, a basic procedure for HDAd production using the Cre-loxP system is presented. Amplification and large-scale production of the vector can be done in both adherent and suspension cell culture systems. Included are protocols for Southern blot analysis to monitor vector amplification, slot blot assay to determine the infectious titer of the purified HDAd, and real-time PCR to detect helper virus contamination in the preparation. PMID- 18428375 TI - An overview of biochemical genetics. AB - Biochemical genetics is focused on the pathophysiology, diagnosis, and treatment of inborn errors of metabolism, which result in potentially toxic accumulation of substrate or secondary metabolites, and/or inhibition of reaction products. Early onset metabolic disorders often present during infancy as acute, life-threatening illnesses; other forms of inborn errors appear later in childhood and are often characterized by progressive neurodegeneration. This unit provides an overview of modern biochemical genetic diagnostic methods used to evaluate a patient's metabolic status via measurement of reaction products or a specific enzyme activity. One such technique, tandem mass spectrometry (MS/MS), has revolutionized population-scale newborn screening for metabolic disorders by providing rapid, accurate, multianalyte quantification on small samples. The combination of expanded newborn screening and novel therapeutic approaches, such as enzyme-replacement therapy, offers significant improvement in the management of metabolic disorders. PMID- 18428376 TI - Metaphase chromosome preparation from cultured peripheral blood cells. AB - Chromosome preparations currently provide the only direct view of the genome as a whole. Although molecular methods allow a more detailed analysis of specific regions of the genome, the study of genetics is not complete without an appreciation of the metaphase cell. The stimulated T cell system described in this unit is the most widely used means of obtaining large numbers of mitotic cells for genetic analyses. Synchronization of the cell cycle in culture is described, combined with direct inhibition of chromosome condensation, to yield longer high-resolution prophase or prometaphase preparations. Such preparations are used for detailed analysis of microdeletions or subtle rearrangements, fine breakpoint analysis, and refined mapping. Microscope slide preparation of mitotic chromosomes from harvested cell culture suspensions is also explained in the support protocol. PMID- 18428377 TI - The BAC resource: tools for array CGH and FISH. AB - Bacterial Artificial Chromosome (BAC) vector clones carrying human DNA were chosen as the intermediate templates for the sequencing of the human genome due to their inherent stability and fidelity to the genome sequence from which they were derived. In this unit, we describe a set of protocols for BAC-based array comparative genomic hybridization (aCGH) that comprise the generation of targets for printing solutions onto glass slides, the subsequent hybridization steps, and CGH analysis of a test sample compared to a reference normal sample. The BAC clones through their sequence allow the extent and gene content of numerical aberrations to be delineated by aCGH, and also provide cytogeneticists with tools for subsequent validation or fine mapping studies. PMID- 18428378 TI - In situ hybridization to metaphase chromosomes and interphase nuclei. AB - In situ hybridization is used to determine the chromosomal map location and the relative order of genes and DNA sequences within a chromosomal band. It can also be used to detect aneuploidy, gene amplification, and subtle chromosomal rearrangements. Fluorescence in situ hybridization (FISH), probably the most widely used method, is described in the first basic protocol. Two support protocols are provided to amplify weak fluorescent signals obtained in FISH. Nonisotopic probes can also be detected by enzymatic reactions using horseradish peroxidase or alkaline phosphatase, as described in alternate protocols. Nonisotopic labeling of DNA probes by nick translation is described in a support protocol. The order of closely spaced FISH probes along chromosomes in interphase nuclei can be determined. A basic protocol for isotopic in situ hybridization (IISH) with (3)H is provided followed by a support protocol for preparation of autoradiographic emulsion. PMID- 18428380 TI - High-resolution FISH analysis. AB - Map order, orientation, and gap or overlap distance of closely linked DNA probes may be determined using fluorescent hybridization to decondensed DNA. The linear arrangement of released chromatin fibers not only simplifies the task of gene ordering, but also provides higher resolution with probes separated by greater distances than can be achieved in FISH with intact interphase nuclei. The Basic Protocol 1 of this unit describes an alkaline lysis procedure for generating free chromatin from cultured cells for FISH analysis. A support protocol describes an empirical approach to optimize conditions for preparation of free chromatin. An Alternate Protocol 1 provides a method for producing free chromatin from cultured lymphocytes with drug treatment. The Basic Protocol 2, high-resolution FISH mapping with free chromatin, is a modification of the method used for FISH mapping of interphase nuclei. PMID- 18428381 TI - Morphology antibody chromosome technique for determining phenotype and genotype of the same cell. AB - The morphology antibody chromosome (MAC) technique is a powerful combination of methods that permits analysis of both phenotype and genotype on a single interphase or mitotic cell as a basis for lineage analysis of neoplastic and normal cells. This unit describes MAC with sequential phenotypic analysis using antibody and an alkaline phosphatase anti-alkaline phosphatase (APAAP) complex and genotypic analysis using in situ hybridization with either enzymatic or fluorescence detection. Alternate methods for phenotypic analysis are also described, which include the use of horseradish peroxidase-conjugated antibodies, fluorochrome-conjugated antibodies, May-Grunwald-Giemsa cytochemical staining, and Sudan black B cytochemical staining. An additional protocol describes G- and C-banding as alternatives to in situ hybridization (ISH) for genotyping MAC specimens. Support protocols describe methods for preparing specimenscytospin preparations, in situ cultures, paraffin-embedded or cryostat sections, and blood and bone marrow smears. Basic Protocol 2 describes a procedure for chromosome painting of previously GTG-banded slides. PMID- 18428382 TI - Human mutation databases. AB - The first part of this unit compares general and locus-specific mutation databases. The second section deals with submitting data. The third part provides guidance for accessing mutation data. The final section offers advice on database construction. PMID- 18428379 TI - Microscopy and image analysis. AB - This unit provides an overview of light microscopy, including objectives, light sources, filters, film, and color photography for fluorescence microscopy and fluorescence in situ hybridization (FISH). Computerized image-analysis systems currently used in clinical cytogenetics are also discussed. PMID- 18428383 TI - Infectious diseases testing. AB - Molecular methods have been applied widely for the diagnosis of infectious diseases. Beginning with solution hybridization in the early 1990s, multiple methods for nucleic acid hybridization and amplification have been introduced into the laboratory for the identification and characterization of microbial pathogens. This unit contains examples of several basic approaches for microbial detection or characterization in the laboratory. Methods in this chapter include automated nucleic acid extraction, direct detection of microbial pathogens, and characterization of pathogens by DNA sequencing or typing. Any of these methods could be customized for the characterization of bacteria, fungi, parasites, or viruses. PMID- 18428384 TI - Techniques for mammalian cell tissue culture. AB - This appendix opens with detailed discussions on the latest principles of sterile technique and preparation of culture media. Step-by-step protocols describe trypsinizing and subculturing monolayer cultures, passaging suspension cultures, freezing and thawing cells, counting cells using a hemacytometer, and preparing cells for transport. PMID- 18428385 TI - Use of LINKAGE programs for linkage analysis. AB - Genetic linkage analysis remains a powerful tool for identifying regions of the genome that may harbor suceptibility loci. This unit describes the traditional approach of calculating two-point and multipoint lod scores when the underlying genetic model is known. Components of the genetic model include whether a trait is dominant, recessive, or codominant, whether it is autosomal or sex linked, and whether there is mutation at the disease gene locus, as well as the disease and marker allele frequencies and the penetrance of the disease allele. The approach to genetic linkage analysis presented in this unit assumes that the markers are loosely spaced, so that there is no linkage disequilibrium between genetic markers. Practical examples and step-by-step guidelines are presented. PMID- 18428386 TI - Detecting mutations in the APC gene in familial adenomatous polyposis (FAP). AB - Hereditary forms of colorectal cancer (CRC) account for up to 5% of total cases. Familial adenomatous polyposis (FAP) is an autosomal dominant condition affecting nearly 1 in 5000 people and accounts for only about 1% of all CRCs. It is characterized by the progressive development of hundreds to thousands of adenomatous colon polyps. The gene associated with FAP (APC) contains 15 exons in the coding region. A scanning approach for large genes is reasonable, but some standard techniques have limited analytical sensitivity. The method described here, using DHPLC as mutation scanning approach for medium-throughput DNA sequence analysis, is largely considered to be the gold standard for point mutation analysis, and can be optimized for high-throughput testing. Detection of deletion and duplication mutations refractory to sequencing have been described using real-time quantitative PCR for dosage analysis. Technical strategies for mutation detection in the APC gene are presented in this chapter. PMID- 18428387 TI - EGF receptor testing for non-small cell lung carcinomas. AB - Non-small cell lung cancer (NSCLC) is one of the most common cancers worldwide. An estimated 170,000 new diagnoses are expected this year. Recently, small molecule inhibitors directed at the EGFR kinase domain were approved for the treatment of advanced stages of NSCLC. Genotyping of the EGFR kinase domain has proven to be a useful marker for predicting who will respond to these novel medications. This unit provides a protocol to perform mutation analysis on the EGFR kinase domain where mutations have been associated with significant responsiveness to these EGFR inhibitors. The protocol includes microdissection of tumor tissue from slides, DNA digestion of these cells, amplifying and sequencing pertinent segments of the EGFR gene, and interpretation of the data. The protocol is designed with appropriate redundancy to eliminate allele dropout and to maximize detection of somatic mutations within the tumor. PMID- 18428388 TI - Biosafety in handling gene transfer vectors. AB - This unit is devoted to safety issues that must be considered when generating and working with vectors for gene therapy. A short description of the relevant regulatory bodies and mechanisms is followed by a general discussion of containment and safety procedures. This is followed by vector-specific guidelines for nine commonly used vector systems, including adenovirus, vaccinia, HSV-1, lentivirus, and nonviral vectors. Recent tragic experiences with retroviral integration near oncogene loci demonstrate the surprise effects of successful gene therapy. The unit also outlines the problems encountered when taking a promising vector to clinical trials. PMID- 18428389 TI - Ex vivo and in vivo gene delivery to the brain. AB - This unit describes methods for grafting genetically modified cells for ex vivo delivery of specific genes into the rat brain and direct delivery of transgenes to brain cells in vivo using recombinant viral vectors. These methods assess the function of a gene in the brain. The ex vivo approach of gene transfer to the nervous system depends on genetic manipulation of cells in vitro prior to grafting of the cells into the brain to enable production of a transgene at physiologically significant levels for a long period of time. This unit also includes procedures for transcardial perfusion to fix the tissue prior to analysis of expression and for sectioning of brains by use of a freezing sledge microtome. Thionine staining of tissue sections is also described. PMID- 18428390 TI - Overview of model systems for the analysis of human disease. AB - The development of animal models of human disease has been crucial in elucidating normal organ biology and pathogenic mechanisms leading to disease states, and in the development of new therapeutic strategies to modify disease. This unit provides an overview of various animal models used in the study of human disease and the factors that need to be considered when selecting a suitable animal model appropriate for the disease under investigation. PMID- 18428391 TI - Yeast as a model for human disease. AB - The sequencing of the human genome promised the identification of disease-causing genes and, subsequently, therapies for those diseases. However, when identifying the genetic basis of a disease, it is not uncommon to discover an abnormal protein whose normal function is unknown. The genetic manipulations required to assign function to genes is often extremely difficult, if not impossible, in human cells. Model organisms have been used to facilitate understanding of gene function because of the ease of genetic manipulations and because many features of eukaryotic physiology have been conserved across phyla. Yeast is a simple eukaryote with a tractable genome, a short generation time, and a large network of researchers who have generated a vast arsenal of research tools. These traits make yeast ideally suited to help reveal the function of genes implicated in human disease. PMID- 18428392 TI - Chromatographic analysis of amino and organic acids in physiological fluids to detect inborn errors of metabolism. AB - This unit describes methods for the preparation of samples for analysis of physiological amino acids and organic acids. Amino acids are analyzed by ion exchange chromatography using an automated system. Organic acids are analyzed by gas-chromatography/mass spectrometry (GC-MS). Analysis of amino and organic acids is necessary to detect and monitor the treatment of many inborn errors of metabolism. PMID- 18428393 TI - Selective isolation of mammalian genes by TAR cloning. AB - Transformation-associated recombination (TAR) cloning provides a unique tool for selective isolation of desired chromosome segments and full-size genes from complex genomes in the form of a circular yeast artificial chromosome (YAC) up to 250 kb in size. The method has a broad application for structural and functional genomics, long-range haplotyping, mutational analysis of gene families, characterization of chromosomal rearrangements, and evolutionary studies. This unit describes a procedure for gene isolation by TAR as well as a method for conversion of YAC-TAR isolates into a bacterial artificial chromosome (BAC) form. PMID- 18428394 TI - Searching the NCBI databases using Entrez. AB - One of the most widely-used interfaces for the retrieval of information from biological databases is the NCBI Entrez system. Entrez capitalizes on the fact that there are pre-existing, logical relationships between the individual entries found in numerous public databases. The existence of such natural connections, mostly biological in nature, argued for the development of a method through which all the information about a particular biological entity could be found without having to sequentially visit and query disparate databases. Two Basic Protocols describe simple, text-based searches, illustrating the types of information that can be retrieved through the Entrez system. An Alternate Protocol builds upon the first Basic Protocol, using additional, built-in features of the Entrez system, and providing alternative ways to issue the initial query. The Support Protocol reviews how to save frequently-issued queries. Finally, Cn3D, a structure visualization tool, is also discussed. PMID- 18428395 TI - DNA mutation detection using denaturing high-performance liquid chromatography (DHPLC). AB - DHPLC is an efficient method for candidate gene scanning with a high level of automation. Single-base substitutions and insertions or deletions of up to 1.5 kb in PCR amplified DNA fragments can be detected. The method exploits the differential retention of homoduplex and heteroduplex DNA species under conditions of partial thermal denaturation. DHPLC provides a useful platform for high-throughput mutation detection and SNP discovery. PMID- 18428396 TI - MLPA and MAPH: sensitive detection of deletions and duplications. AB - The detection of quantitative changes in genomic DNA, i.e., deletions and duplications or so called Copy Number Variants (CNV), is an important element of a complete mutation screening strategy. However, because of practical difficulties, screening for quantitative changes in genomic DNA is often ignored. Hitherto, the techniques available were technically challenging and laborious and thus too costly to be applied on a routine basis. The development of MAPH (Multiplex Amplifiable Probe Hybridization) and more recently MLPA (Multiplex Ligation-dependent Probe Amplification) have revived interest in the detection of deletions and duplications, primarily due to the simplicity and flexibility of these two approaches. Compared to previous technologies, e.g., Southern blotting, fluorescence in situ hybridization (FISH), quantitative PCR (qPCR), and breakpoint PCR, they have some clear advantages, including high resolution, high throughput, amenability to multiplexing, and simplicity. PMID- 18428397 TI - Overview of molecular genetic diagnosis. AB - This unit describes the two approaches for molecular genetic diagnosis, direct mutation analysis and linkage studies. Direct mutation analysis is possible if the gene responsible for a disorder has been cloned and techniques exist to detect mutations. Genetic linkage analysis is used when polymorphic markers are available that are closely linked to a disease gene but the gene itself has not been cloned, or when it is difficult or impossible to detect mutations in the gene. For direct mutation analysis, the unit offers a review of the types of mutations that can be detected and the strategies that can be employed for their detection. The unit then describes the types of disorders that are best suited for linkage-based analysis and offers guidance for interpreting the data. PMID- 18428398 TI - Internet resources in Medical Genetics. AB - This unit presents an overview of the most commonly used, Web-based information resources for (1) clinicians seeking to apply molecular genetic testing to patient care (see GeneTests) or to understand the molecular basis of inherited diseases (see OMIM), (2) consumers seeking advocacy information (see Genetic Alliance) or scientific and management information (see The Genetics Home Reference, The Genetics Education Center, The Family Village, NOAH, NORD, and Genes and Diseases), and (3) genetics professional societies. PMID- 18428399 TI - Overview of pharmacogenetics. AB - Pharmacogenetics is the study of relationships between genetic variation and inter-individual differences with respect to drug response. As the field has matured over the past 15 years, a remarkable diversity of pathways, variation types, and mechanisms have been found to be relevant pharmacogenetic factors. Today, pharmacogenetics is becoming more important in pharmacology for target validation, lead optimization, and understanding of idiosyncratic toxicity. This unit provides an overview of the history of pharmacogenetics and current research applications in drug discovery, as well as a discussion of research quality issues relevant for human subjects research in the pharmacogenetics laboratory. PMID- 18428400 TI - Multiplex PCR for identifying DMD gene deletions. AB - The identification of dystrophin as the defective protein in patients with Duchenne and Becker muscular dystrophies (DMD and BMD) has allowed the development of sensitive and specific tests to establish a diagnosis and to aid in genetic counseling and prenatal diagnosis. The Basic Protocol describes three complementary multiplex PCR assays that detect 26 dystrophin gene exons. The multiplex nature of these assays allows the detection of up to ten different exons in a single reaction. At least one of these exons is missing in >95% of deletions. The Support Protocol describes preparation and storage of stock PCR reaction mixes with primers for each of the three diagnostic assays. The Alternate Protocol is a modification of the Basic Protocol for radioactive detection of duplications in males and deletions in carrier females. PMID- 18428401 TI - Quantitation of DNA and RNA with absorption and fluorescence spectroscopy. AB - Quantitation of nucleic acids is a fundamental tool in molecular biology that requires accuracy, reliability, and the use of increasingly smaller sample volumes. This unit describes the traditional absorbance measurement at 260 nm and three more sensitive fluorescence techniques, as well as three microvolume methods that use fiber optic technology in specialized cells or instrumentation. These procedures allow quantitation of DNA solutions ranging from 1 pg/microl to 50 mg/ml. PMID- 18428402 TI - Population-based case-control association studies. AB - This unit provides an overview of the design and analysis of population-based case-control studies of genetic risk factors for complex disease. Considerations specific to genetic studies are emphasized. The unit reviews basic study designs, differentiating case-control studies from others, discusses selection of genetic markers for use in studies, introduces basic methods of analysis of case-control data, and discusses measures of association and impact. Controlling for confounding (including population stratification), consideration of multiple loci, and haplotype analysis are briefly discussed. Readers are referred to basic texts on epidemiology for more details on general conduct of case-control studies. PMID- 18428403 TI - Overview of genetic diagnosis in cancer. AB - Both cytogenetic and molecular genetic studies can contribute to the management of patients with cancer. In some cases genetic markers are specific to particular tumor types and are useful in diagnosis. This can be helpful in distinguishing histologically similar tumors that may respond differently to treatment and can sometimes be of prognostic value. Genetic markers can also be tools for following the response of a tumor to therapy, providing a sensitive means of detection of relapse. This introductory unit considers some of the types of genetic changes that occur in association with malignancies and major approaches used in their detection. PMID- 18428404 TI - Analysis of expression data: an overview. AB - After providing a brief introduction to microarray chips and experimental details, this overview discusses analysis techniques. Data analysis from microarray experiments generally involves two parts: acquiring and normalizing the data, and interpreting it. This unit focuses mostly on the latter, as it is less technology-specific. PMID- 18428405 TI - Serial analysis of gene expression (SAGE): experimental method and data analysis. AB - This unit provides a protocol for performing serial analysis of gene expression (SAGE). SAGE involves the generation of short fragments of DNA, or tags, from a defined point in the sequence of all cDNAs in the sample analyzed. This short tag, because of its presence in a defined point in the sequence, is typically sufficient to uniquely identify every transcript in the sample. SAGE allows one to generate a comprehensive profile of gene expression in any sample desired from as little as 100,000 cells or 1 microg of total RNA. SAGE generates absolute, rather than relative, measurements of RNA abundance levels, and this fact allows an investigator to readily and reliably compare data to those produced by other laboratories, making the SAGE data set increasingly useful as more data is generated and shared. Software tools have also been specifically adapted for SAGE tags to allow cluster analysis of both public and user-generated data. PMID- 18428406 TI - Production and titration of lentiviral vectors. AB - Lentiviral vectors have emerged over the last decade as powerful, reliable and safe tools for stable gene transfer in a wide variety of mammalian cells. Unlike other vectors derived from oncoretroviruses, they allow for stable gene delivery into most nondividing primary cells. This is why LVs are becoming useful and promising tools for future gene and cell therapy approaches. Lentivectors (LVs) derived from HIV-1 have gradually evolved to display many desirable features aimed at increasing both their safety and their versatility. These latest designs are reviewed in this unit. This unit also describes protocols for production and titration of LVs that can be implemented in a research laboratory setting, with an emphasis on standardization to improve transposability of results between laboratories. PMID- 18428407 TI - Production of vesicular stomatitis virus G glycoprotein (VSV-G) pseudotyped retroviral vectors. AB - Retrovirus pseudotype is defined as the genome of one retrovirus encapsidated by the envelope protein of a second virus. The host range of the pseudotype is that of the virus donating the envelope protein. Two procedures that use 293GP cells, which are derived from human kidney 293 cells, are described here. The first is based on the high transient transfection efficiency of 293 cells. The retroviral construct and an expression plasmid for VSV-G are co-transfected into 293GP cells that stably express MLV gag and pol proteins. Transiently generated virus is then harvested during consecutive days following DNA transfection. The second procedure involves stable 293GP cell lines containing the VSV-G gene under the control of a promoter whose activity is regulated by tetracycline. Cell lines containing the retroviral vector of interest are established under noninduced conditions. Infectious virus can be harvested following the induction of VSV-G expression in these cell lines. PMID- 18428408 TI - Production of recombinant adeno-associated viral vectors. AB - Adeno-associated virus is a nonpathogenic human virus that has been developed into a gene-delivery vector due to its high efficiency of infection for many different cell types and its ability to persist and lead to long-term gene expression. This unit describes efficient methods to generate high-titer, research-grade, adenovirus-free recombinant single-stranded and self complementary adeno-associated virus in various serotypes, along with methods to quantify the viral vectors. PMID- 18428410 TI - Detection of hypo-N-glycosylation using mass spectrometry of transferrin. AB - Many congenital disorders of glycosylation (CDG) can be diagnosed by observing the extent of glycosylation of the abundant serum glycoprotein transferrin (Trf). Trf is an N-glycosylated protein with two asparagine glycation sites. CDG types I are those genetic defects which occur prior to transfer of the complex oligosaccharide to the acceptor asparagine in the cotranslated polypeptide chain. CDG Ia constitutes by far the most frequent form of CDG and is the result of mutations in the phosphomannomutase gene. CDG Ia and the Ib subtype (Phosphomannoisomerase deficiency) result in low cellular mannose-1-phosphate levels, a required precursor for oligosaccharide assembly in the endoplasmic reticulum. The deficiency in oligosaccharides with branched mannose structures is thereafter expressed by the appearance of glycoproteins with unoccupied N glycosylation sites (hypoglycosylation). Currently, there have been at least 11 Type I defects, type Ia being by far the most frequently occurring. Most, if not all type I defects result in unoccupied N-glycation sites. Hypoglycosylated Trf, also known as carbohydrate-deficient Trf (CDT), can be detected using mass spectrometry (MS) to measure the masses of the serum Trf. The methods for sample preparation using affinity chromatography and MS analysis are described in this unit. PMID- 18428409 TI - Quantification of creatine and guanidinoacetate using GC-MS and LC-MS/MS for the detection of cerebral creatine deficiency syndromes. AB - Inherited defects in creatine biosynthesis and cellular uptake are neurometabolic disorders characterized by seizures, developmental delay, mental retardation, autistic-like behavior, and creatine deficiency in the brain. Metabolic screening of these disorders is possible using analytical techniques that quantify creatine and its precursor guanidinoacetate in urine, plasma, or cerebrospinal fluid (CSF). Elevated creatine in urine is suggestive of a deficiency of the X-linked creatine transporter, SLC6A8. Decreased or elevated levels of guanidinoacetate in urine, plasma, or CSF suggest deficiencies of the creatine biosynthetic enzymes, arginine:glycine amidinotransferase (AGAT) or guanidinoacetate methyltransferase (GAMT), respectively. This unit describes three stable isotope dilution-mass spectrometric methods for analyzing creatine and guanidinoacetate. Gas chromatography/mass spectrometry with negative-ion chemical ionization is a highly sensitive technique, suitable for detection of low analyte levels resulting from AGAT deficiency and in CSF. The two liquid chromatography-tandem mass spectrometric approaches are amenable to high-throughput screening and have simple sample preparation requirements. PMID- 18428411 TI - Conversion Technology for the separation of maternal and paternal copies of any autosomal chromosome in somatic cell hybrids. AB - Conversion Technology (CT) is a streamlined version of somatic cell hybrid preparation that was developed to be amenable for use as a mutation detection method and as a diagnostic tool for individual patients. It is also a powerful research tool. There are two broad categories of potential applications for CT: research applications in gene mapping and identification, and clinical applications in the detection of disease-causing mutations in individual patients. CT may emerge as the gold standard for both mutation detection and haplotyping. It will likely prove valuable in mutation detection due to its ability to detect mutations that are not amenable to detection by sequencing (e.g., deep intron or promoter mutations). It will likely prove valuable in gene identification projects because genotyping of haploid chromosomes yields unequivocal haplotypes. PMID- 18428412 TI - Human artificial chromosome assembly by transposon-based retrofitting of genomic BACs with synthetic alpha-satellite arrays. AB - The development of methodologies for the rapid assembly of synthetic alpha satellite arrays recapitulating the higher-order periodic organization of native human centromeres permits the systematic investigation of the significance of primary sequence and sequence organization in centromere function. Synthetic arrays with defined mutations affecting sequence and/or organization may be evaluated in a de novo human artificial chromosome assay. This unit describes strategies for the assembly of custom built alpha-satellite arrays containing any desired mutation as well as strategies for the construction and manipulation of alpha satellite-based transposons. Transposons permit the rapid and reliable retrofitting of any genomic bacterial artificial chromosome (BAC) with synthetic alpha-satellite arrays and other functional components, thereby facilitating conversion into BAC-based human artificial chromosome vectors. These techniques permit identification and optimization of the critical parameters underlying the unique ability of alpha-satellite DNA to facilitate de novo centromere assembly, and they will establish the foundation for the next generation of human artificial chromosome vectors. PMID- 18428413 TI - Large-scale BAC clone restriction digest fingerprinting. AB - Restriction digest fingerprinting is a common method for characterizing large insert genomic clones, e.g., bacterial artificial chromosome (BAC), P1 artificial chromosome (PAC) and Fosmid clones. This clone fingerprinting method has been widely applied in the construction of clone-based physical maps, which have been used as positional cloning resources as well as to support directed and genome wide sequencing efforts. This unit describes a robust, large-scale procedure for generation of agarose gel-based clone fingerprints from BAC clones. PMID- 18428414 TI - Fosmid libraries for genomic structural variation detection. AB - Fosmid libraries have demonstrated their utility for a number of applications. These include filling gaps between BACs and small insert libraries in sequence assemblies, performing hybridization/screening studies to isolate functional elements within the genome (Vergin et al., 1998), and detecting insertions, deletions, and rearrangements in structural variation studies (Tuzun et al., 2005). This unit covers the basic methodologies for the construction of fosmid libraries with tight insert sizes suitable for these applications. Basic Protocol 1 covers the shearing, size selection, and recovery of DNA from a pulsed-field gel. Basic Protocol 2 covers the cloning of insert DNA into the fosmid vector, packaging of DNA into infective phage particles, and the infection/transformation of bacteria. A commentary section is provided, which outlines many of the critical parameters involved in fosmid library construction, along with some additional background information and a section discussing anticipated results. PMID- 18428415 TI - Using BLAST for performing sequence alignment. AB - BLAST is a widely used genetic sequence comparison program developed at the National Center for Biotechnology Information (NCBI). In this unit, three Basic Protocols and one Support Protocol are provided for general-purpose BLAST searches on the NCBI and ENSEMBL Web-accessible BLAST servers. Key parameters affecting how the search algorithm works are reviewed, with advice on modifying search parameters for specific situations. Many other public and private Web sites offer BLAST interfaces which may differ from those described in this unit, but the general principles will be similar. The Support Protocol describes how to obtain sequences in various formats from NCBI for use in BLAST searches. It is emphasized that no algorithm can be a substitute for biological understanding; performing a BLAST search takes only a few minutes but understanding the implications of the results takes much longer. PMID- 18428416 TI - Quality assurance and quality control in clinical cytogenetics. AB - The goal of any clinical laboratory should be to provide patients with the most accurate test results possible. This is accomplished through various overlapping programs that continuously monitor and optimize all aspects of a test, including decisions by the laboratory to offer a test, the decision of providers to request the test, the testing itself, and the reporting of results to the referral source and patient. The levels at which test performance and accuracy can be optimized are encompassed under quality control (QC) and quality assurance (QA). The monitoring of QC and QA problems allows for the integration of these parameters into a total quality management program. This unit reviews QC and QA guidelines, in addition to discussing how to establish a quality assurance program. PMID- 18428417 TI - Preparation of cells from formalin-fixed, paraffin-embedded tissue for use in fluorescence in situ hybridization (FISH) experiments. AB - Numerical and structural chromosome abnormalities can be accurately detected in cells from archived tissues using fluorescence in situ hybridization (FISH). This unit describes two common approaches to performing FISH in formalin-fixed, paraffin-embedded tissue. The first approach utilizes 4 to 6 microm tissue sections in cases for which preserving tissue morphology is necessary, and the second involves extraction of intact nuclei from 50 microm tissue sections. To interpret FISH results using 4 to 6 microm sections, an adequate number of nuclei must be evaluated to perform statistical analysis. Evaluation of 30 to 50 nuclei from the single cell suspension generally gives an interpretable result. PMID- 18428418 TI - Laboratory safety. AB - The pursuit of scientific knowledge has been considerably advanced by the use of a wide variety of hazardous substances, including radioisotopes, hazardous chemicals, and biohazards. This appendix addresses each of these classes of hazards and primary safety precautions that need to be considered. PMID- 18428419 TI - Calculation and use of the Hardy-Weinberg model in association studies. AB - Hardy-Weinberg equilibrium (HWE) is an important tool for understanding population structure. If certain assumptions are met, genotype and allele frequencies can be estimated from one generation to the next. In genetic association studies, HWE principles have been applied to detect genotyping error and disease susceptibility loci. The focus of this unit is to review the key principles and assumptions of HWE. There is a brief discussion on how the significance of HWE is tested, and a review of current applications of HWE in association studies. The applications discussed include estimating penetrance, evaluating genotyping errors, testing for population stratification, and testing for association. PMID- 18428420 TI - p53 Testing for Li-Fraumeni and Li-Fraumeni-like syndromes. AB - Li-Fraumeni Syndrome (LFS; OMIM #151623) is an autosomal dominant cancer predisposition syndrome characterized by early onset tumors including sarcomas, breast cancer, leukemia, brain tumors, and adrenocortical carcinoma. Li-Fraumeni syndrome is primarily attributed to germline mutations in the p53 tumor suppressor gene, which encodes tumor protein 53. In addition to germline p53 mutations, the p53 gene is the most commonly mutated gene in human cancers, with as much as 50% of tumors containing somatic p53 mutations. This unit provides a protocol to perform germline mutation analysis of the p53 gene. The protocol includes steps for amplification and sequencing of the entire coding region of the p53 gene (exons 2 to 11). The protocol was designed for detecting germline alterations from DNA extracted from blood; however, with some additional optimization, it could also be used to detect somatic mutations in DNA extracted from tumors. PMID- 18428421 TI - The Catalogue of Somatic Mutations in Cancer (COSMIC). AB - COSMIC is currently the most comprehensive global resource for information on somatic mutations in human cancer, combining curation of the scientific literature with tumor resequencing data from the Cancer Genome Project at the Sanger Institute, U.K. Almost 4800 genes and 250000 tumors have been examined, resulting in over 50000 mutations available for investigation. This information can be accessed in a number of ways, the most convenient being the Web-based system which allows detailed data mining, presenting the results in easily interpretable formats. This unit describes the graphical system in detail, elaborating an example walkthrough and the many ways that the resulting information can be thoroughly investigated by combining data, respecializing the query, or viewing the results in different ways. Alternate protocols overview the available precompiled data files available for download. PMID- 18428422 TI - Human hematopoietic cell culture, transduction, and analyses. AB - This unit provides methods for introducing genes into human hematopoietic progenitor cells. The Basic Protocol describes isolation of CD34(+) cells, transduction of these cells with a retroviral vector on fibronectin-coated plates, assaying the efficiency of transduction, and establishing long-term cultures. Support protocols describe methods for maintenance of vector-producing fibroblasts (VPF) and supernatant collection from these cells, screening medium components for the ability to support hematopoietic cell growth, and establishing colonies from long-term cultures. Other protocols provide PCR-based methods to analyze individual colonies for transduction, methods to analyze cells harvested from long-term cultures, and procedures for freezing and thawing of hematopoietic cells. PMID- 18428423 TI - Diagnosis of inherited disorders of galactose metabolism. AB - Galactose metabolism occurs through an evolutionarily conserved pathway in which galactose and uridine diphosphoglucose are converted to glucose-1-phosphate and uridine diphosphogalactose through the action of three sequential enzymes: galactokinase (GALK, EC 2.7.1.6), galactose-1-phosphate uridyltransferase (GALT, EC 2.7.7.12), and uridine phosphogalactose 4'-epimerase (GALE, EC 5.1.3.2). Inborn errors of galactose metabolism occur with impaired activity for each of the enzymes. Classical galactosemia is the most common and the most severe of these diseases and is caused by deficiency of the GALT enzyme, affecting from approximately 1 in 10,000 to 1 in 30,000 live births. Deficiency of GALE is the rarest of the three diseases. Assays for galactitol and galactose-1-phosphate and methods for assaying enzyme activities of GALT, GALK, and GALE are provided here. Interpretation of diagnostic results for screen-positive newborns or symptomatic patients, as well as therapeutic interventions based on biochemical phenotype and molecular genotype, are also included as decision trees. PMID- 18428424 TI - Genotyping using the TaqMan assay. AB - The 5'-nuclease allelic discrimination assay, or TaqMan assay, is a PCR-based assay for genotyping single nucleotide polymorphisms (SNPs). The region flanking the SNP is amplified in the presence of two allele-specific fluorescent probes. The probes do not fluoresce in solution because of a quencher at the 3' end. The presence of two probes allows the detection of both alleles in a single tube. Moreover, because probes are included in the PCR, genotypes are determined without any post-PCR processing, a feature that is unavailable with most other genotyping methods. This unit describes probe and primer design and PCR conditions. PMID- 18428425 TI - Selection of a platform for mutation detection. AB - New mutation detection technologies must keep pace by becoming more cost effective while offering improved technical sensitivity and higher throughput capacity. In recent years, the number of mutation detection platforms available to the clinical researcher has grown to a point where it is difficult to keep track of all available options as well as their benefits and pitfalls. This unit provides an entry point for a variety of researchers who wish to analyze samples for known or novel mutations and need to determine which platform is most suited for their particular needs. A practical guide is provided in this unit, including a brief overview, information on assay parameters, design and cost considerations, as well as platform flexibility and scalability of the assay. Although the focus here is on applications involving human disease, many of these platforms can be easily adapted to the study of other organisms. PMID- 18428426 TI - Mutation detection using automated fluorescence-based sequencing. AB - The development of high-throughput DNA sequencing techniques has made direct DNA sequencing of PCR-amplified genomic DNA a rapid and economical approach to the identification of polymorphisms that may play a role in disease. Point mutations as well as small insertions or deletions are readily identified by DNA sequencing. The mutations may be heterozygous (occurring in one allele while the other allele retains the normal sequence) or homozygous (occurring in both alleles). Sequencing alone cannot discriminate between true homozygosity and apparent homozygosity due to the loss of one allele due to a large deletion. In this unit, strategies are presented for using PCR amplification and automated fluorescence-based sequencing to identify sequence variation. The size of the project and laboratory preference and experience will dictate how the data is managed and which software tools are used for analysis. A high-throughput protocol is given that has been used to search for mutations in over 200 different genes at the Harvard Medical School - Partners Center for Genetics and Genomics (HPCGG, http://www.hpcgg.org/). PMID- 18428427 TI - Retrospective analysis of 65 Chinese children with acute promyelocytic leukemia: a single center experience. AB - BACKGROUND: There are very limited data reported about childhood acute promyelocytic leukemia (APL), especially with arsenic trioxide (As2O3) treatment. We review the clinical course and treatment outcome of 65 children APL. PROCEDURE: Between January 1999 and December 2005, 65 children (<18 years) with newly diagnosed APL were treated. RESULTS: Three groups of patients were identified according to the different induction regimens. Forty patients were given ATRA alone (group 1, G1), 8 patients were given As2O3 alone (group 2, G2), 15 patients (group 3, G3) were treated with combination of ATRA and As2O3. Two patients received cytosine arabinoside and died at days 2 and 4, respectively. The CR rate of G1 was 95% (38/40). The overall CR rate of G2+G3 was 91.3% (21/23). Toxicity of ATRA and As2O3 were minimal and recoverable during induction. Five children had a hematologic relapse and two developed molecular relapse. Five of them were successfully treated with ATRA alone or combined with As2O3. The 5-year estimate of EFS, DFS and OS between G1 and G2+G3 has no difference. CONCLUSION: As2O3 is an effective and well tolerable therapy for children with APL and it may be used in those who have dose limiting side effects of ATRA, but also for those with newly diagnosed or relapsed APL. PMID- 18428428 TI - Analysis of multiple SNPs in a candidate gene or region. AB - We consider the analysis of multiple single nucleotide polymorphisms (SNPs) within a gene or region. The simplest analysis of such data is based on a series of single SNP hypothesis tests, followed by correction for multiple testing, but it is intuitively plausible that a joint analysis of the SNPs will have higher power, particularly when the causal locus may not have been observed. However, standard tests, such as a likelihood ratio test based on an unrestricted alternative hypothesis, tend to have large numbers of degrees of freedom and hence low power. This has motivated a number of alternative test statistics. Here we compare several of the competing methods, including the multivariate score test (Hotelling's test) of Chapman et al. ([2003] Hum. Hered. 56:18-31), Fisher's method for combining P-values, the minimum P-value approach, a Fourier-transform based approach recently suggested by Wang and Elston ([2007] Am. J. Human Genet. 80:353-360) and a Bayesian score statistic proposed for microarray data by Goeman et al. ([2005] J. R. Stat. Soc. B 68:477-493). Some relationships between these methods are pointed out, and simulation results given to show that the minimum P value and the Goeman et al. ([2005] J. R. Stat. Soc. B 68:477-493) approaches work well over a range of scenarios. The Wang and Elston approach often performs poorly; we explain why, and show how its performance can be substantially improved. PMID- 18428429 TI - Quantifying the contribution of genetic variants for survival phenotypes. AB - Particularly in studies based on population representative samples, it is of major interest what impact a genetic variant has on the phenotype of interest, which cannot be answered by mere association estimates alone. One possible measure for quantifying the phenotype's variance explained by the genetic variant is R(2). However, for survival outcomes, no clear definition of R(2) is available in the presence of censored observations. We selected three criteria proposed for this purpose in the literature and compared their performance for single nucleotide polymorphism (SNP) data through simulation studies and for mortality data with candidate SNPs in the general population-based KORA cohort. The evaluated criteria were based on: (1) the difference of deviance residuals, (2) the variation of individual survival curves, and (3) the variation of Schoenfeld residuals. Our simulation studies included various censoring and genetic scenarios. The simulation studies revealed that the deviance residuals' criterion had a high dependence on the censoring percentage, was generally not limited to the range [0; 1] and therefore lacked interpretation as a percentage of explained variation. The second criterion (variation of survival curves) hardly reached values above 60%. Our requirements were best fulfilled by the criterion based on Schoenfeld residuals. Our mortality data analysis also supported the findings in simulation studies. With the criterion based on Schoenfeld residuals, we recommend a powerful and flexible tool for genetic epidemiological studies to refine genetic association studies by judging the contribution of genetic variants to survival phenotype. PMID- 18428430 TI - Comparison of gallium and PET scans at diagnosis and follow-up of pediatric patients with Hodgkin lymphoma. AB - BACKGROUND: Positron emission tomography (PET) and gallium scans facilitate diagnosis and staging, evaluation of response to therapy, and monitoring for relapse in Hodgkin lymphoma (HL), but have not been compared in pediatric HL. PROCEDURE: We performed concurrent PET and gallium scans on 44 pediatric HL patients at diagnosis, early response, off chemotherapy, and off-therapy evaluations. PET and gallium scans were compared to each other and to computed tomography (CT) alone to determine whether either modality led to a change in stage or modified the results of the early response evaluation, which was used to determine the radiation dose. RESULTS: PET upstaged four patients at diagnosis (2 from stage I to II, one II to III, and one III to IV), but did not lead to a change in therapy in any of them. It changed response category in two patients at early response evaluation, leading to a change in radiation dose for 1 patient (25.5 Gy instead of 15 Gy to the spleen). Gallium did not change the stage of treatment for any patient. The negative predictive values for eventual lymphoma relapse of PET and gallium scans at off therapy were 89% and 83%, respectively; the positive predictive value of PET at off therapy is 29%. CONCLUSION: PET appears to be superior to gallium in pediatric HL; future studies will determine the optimal timing of PET to assess early response and the utility of quantitative interpretation of the avidity of specific nodal sites. PMID- 18428431 TI - Methadone analgesia for children with advanced cancer. AB - BACKGROUND: Methadone is frequently used in the treatment of adults with advanced cancer. A criticism of relevant research is the use of single or fixed doses, which does not reflect use in clinical practice. Literature about use of methadone in the treatment of pediatric patients is limited to case reports. The objective of this study is to describe methadone use as primary opioid analgesic for advanced pediatric cancer over a 6.5-year period. PROCEDURE: All 17 patients who received methadone as their primary opioid analgesic through the Northern Alberta Children's Cancer Program from January 2000 to June 2007 were included. Children who received combination opioid therapy were excluded. RESULTS: Rotation to methadone was usually by a complete switch from primary opioid. Conversion ratios of morphine equivalent daily dose (MEDD)/methadone daily dose (TMDD) ranged widely from 1:2 in one patient with sudden pain crisis just prior to death, to 60:1 in a patient who had been treated with opioids for months. Methadone was used for a total of 925 patient-days. There were no significant adverse events in any patient, and all but one patient remained on methadone until the time of their death. Clinically, the effectiveness of analgesia clearly improved at time of conversion in 16 patients. CONCLUSION: With close monitoring, methadone therapy can be done safely in pediatric oncology patient populations in both inpatient and outpatient settings. Our experience suggests improvement in analgesia with the use of methadone, with 16 patients remaining on methadone until they died. PMID- 18428432 TI - The role of hematopoietic stem cell transplantation with relapsed or primary refractory childhood B-cell non-Hodgkin lymphoma and mature B-cell leukemia: a retrospective analysis of enrolled cases in Japan. AB - BACKGROUND: There have been excellent treatment results for children with B-cell non-Hodgkin lymphoma (B-NHL) and mature B-cell leukemia (B-ALL) in the last few decades. However, a small subset of relapsed or refractory patients, after first line therapy, still have a poor prognosis. PROCEDURE: Thirty-three patients with relapsed or primary refractory B-NHL/B-ALL among 327 newly diagnosed patients between 1996 and 2004 were analyzed retrospectively. RESULTS: After salvage therapy, 18 patients were chemotherapy-sensitive and 15 patients suffered from progression. Among 18 patients who had a chemotherapy-sensitive disease, 4 of 5 patients who underwent hematopoietic stem cell transplantation (HSCT) during remission survived without progression, while 3 of 12 patients who did not receive HSCT were alive without disease progression. Fifteen patients never sensitive to salvage therapy died. CONCLUSIONS: Patients with relapsed/primary refractory B-NHL/B-ALL have a poor prognosis with current treatment approaches, while the patients sensitive to salvage therapy have a respectable chance to achieve a sustained complete second remission with HSCT. PMID- 18428433 TI - Optimization of transfection. AB - The single most important factor in optimizing transfection efficiency is selecting the proper transfection protocol. This overview discusses the most commonly used procedures: calcium phosphate-mediated gene transfer, DEAE-dextran mediated gene transfer, electroporation, and liposome-mediated transfection. Fusion techniques such as protoplast fusion and microinjection may also be considered. PMID- 18428434 TI - Calcium phosphate transfection. AB - This unit presents two methods of calcium phosphate-based eukaryotic cell transfection that can be used for both transient and stable transfections. In these protocols, plasmid DNA is introduced to monolayer cell cultures via a precipitate that adheres to the cell surface. A HEPES-buffered solution is used to form a calcium phosphate precipitate that is directly layered onto the cells. For some cells, shocking the cells with glycerol or DMSO improves transfection efficiency. In the alternate high-efficiency method, a BES-buffered system is used that allows the precipitate to form gradually in the medium and then drop onto the cells. While the alternate method is particularly efficient for stable transformation of cells with circular plasmid DNA, both protocols yield similar results for transformation with linear plasmid or genomic DNA, or for transient expression. PMID- 18428435 TI - Transfection using DEAE-dextran. AB - Transfection of cultured mammalian cells using diethylaminoethyl (DEAE) dextran/DNA can be an attractive alternative to other transfection methods in many circumstances. The major advantages of the technique are its relative simplicity and speed, limited expense, and remarkably reproducible interexperimental and intraexperimental transfection efficiency. Disadvantages include inhibition of cell growth and induction of heterogeneous morphological changes in cells. Furthermore, the concentration of serum in the culture medium must be transiently reduced during the transfection. In general, DEAE-dextran DNA transfection is ideal for transient transfections with promoter/reporter plasmids in analyses of promoter and enhancer functions, and is suitable for overexpression of recombinant protein in transient transfections or for generation of stable cell lines using vectors designed to exist in the cell as episomes. This unit presents a general description of DEAE-dextran transfection, as well as two more specific protocols for typical experimental applications. The basic protocol is suitable for transfection of anchorage-dependent (attached) cells. For cells that grow in suspension, electroporation or lipofection is usually preferred, although DEAE-dextran-mediated transfection can be used. PMID- 18428436 TI - Transfection by electroporation. AB - Electroporation--the use of high-voltage electric shocks to introduce DNA into cells--can be used with most cell types,yields a high frequency of both stable transformation and transient gene expression,and,because it requires fewer steps,can be easier than alternate techniques. In this unit,the basic protocol describes the electroporation of mammalian cells and an alternate protocol outlines modifications for preparation and transfection of plant protoplasts. PMID- 18428437 TI - Transfection of cultured eukaryotic cells using cationic lipid reagents. AB - The development of high-efficiency methods for the introduction of functional genetic material into eukaryotic cells using cationic lipid-based transfection reagents has accelerated biology research in studies of gene expression, control of cell growth, and cell lineage. In this unit, DNA transfection is described for adherent mammalian cells (both cells and primary cultures) along with an alternate procedure for enhanced transfection. A protocol is also described for transfection of suspension cells (lymphoid, myeloid and leukemic-derived cells). In addition, transfection of RNA into adherent mammalian cells and DNA transfection into insect cells are presented. Importantly, a detailed procedure is described for optimization of reagents and transfection conditions. PMID- 18428438 TI - Purification and concentration of DNA from aqueous solutions. AB - This unit presents basic procedures for manipulating solutions of single- or double-stranded DNA through purification and concentration steps. These techniques are useful when proteins or solute molecules need to be removed from aqueous solutions, or when DNA solutions need to be concentrated for reasons of convenience. The Basic Protocol, using phenol extraction and ethanol (or isopropanol) precipitation, is appropriate for the purification of DNA from small volumes (<0.4 ml) at concentrations lower than 1 mg/ml. Three support protocols outline methods to buffer the phenol used in extractions, concentrate DNA using butanol, and extract residual organic solvents with ether. An alternative to these methods is nucleic acid purification using glass beads and this is also presented. These protocols may also be used for purifying RNA. The final two alternate protocols provide modifications to the basic protocol that are used for concentrating RNA and extracting and precipitating DNA from larger volumes and from dilute solutions, and for removing low-molecular-weight oligonucleotides and triphosphates. PMID- 18428439 TI - Preparation of genomic DNA from mammalian tissue. AB - There are a number of different procedures for the preparation of genomic DNA. They all start with some form of cell lysis, followed by deproteination and recovery of DNA. The main differences between various approaches lie in the extent of deproteination and in molecular weight of the DNA produced. The isolation procedure described here is relatively brief and relies on the powerful proteolytic activity of proteinase K combined with the denaturing ability of the ionic detergent SDS. PMID- 18428440 TI - Preparation of RNA from tissues and cells. AB - Most procedures for isolating RNA from eukaryotic cells involve lysing and denaturing cells to liberate total nucleic acids. Additional steps are then required to remove DNA. The first basic protocol describes hot phenol extraction of RNA; the method eliminates or minimizes DNA contamination by the shearing of DNA. The second basic protocol allows rapid preparation of total cytoplasmic RNA by using a nonionic detergent to lyse the plasma membrane, leaving the nuclei intact. The nuclei and hence the bulk of the cellular DNA are then removed with a simple brief centrifugation. A guanidinium thiocyanate protocol describes the separation of RNA from other cellular macromolecules in a guanidinium lysate using a CsCl step gradient. A protocol is also provided for isolation of poly(A(+)) mRNAs from total RNA. PMID- 18428441 TI - Preparation of bacterial plasmid DNA. AB - The protocols in this unit describe methods for preparing bacterial plasmid DNA free from chromosomal DNA. The first is an alkaline lysis miniprep suitable for screening a moderate number of bacterial colonies by restriction endonuclease cleavage and agarose gel electrophoresis. The second is the first step to producing large amounts (milligrams) of plasmid DNA and is also based on alkaline lysis of the bacterial cells. The crude lysate generated in this protocol can be further purified by centrifugation using CsCl/ethidium bromide (CsCl/EtBr) equilibrium density gradients. Three support protocols provide information on how to grow overnight and larger cultures of bacteria, and how to monitor bacterial growth with a spectrophotometer. Other methods, some relying on commercially available ion-exchange columns, are discussed in the commentary. PMID- 18428442 TI - Quantitation of DNA and RNA with absorption and fluorescence spectroscopy. AB - Reliable quantitation of nanogram and microgram amounts of DNA and RNA in solution is essential to researchers in molecular biology. In addition to the traditional absorbance measurements at 260 nm, two more sensitive fluorescence techniques are presented below. These three procedures cover a range from 5 to 10 ng/ml DNA to 50 mg/ml DNA. PMID- 18428443 TI - Introduction of plasmid DNA into cells. AB - This appendix presents a procedure for transformation using calcium chloride, and also an alternate procedure for one-step preparation and transformation of competent cells. PMID- 18428444 TI - Digestion of DNA with restriction endonucleases. AB - Restriction endonucleases recognize short DNA sequences and cleave double stranded DNA at specific sites within or adjacent to the recognition sequences. Restriction endonuclease cleavage of DNA into discrete fragments is one of the most basic procedures in molecular biology. This appendix describes restriction endonucleases and their properties. PMID- 18428446 TI - Common stock solutions, buffers, and media. PMID- 18428445 TI - Agarose gel electrophoresis. AB - This appendix presents a protocol for separating and purifying DNA fragments between 0.5 and 25 kb. A support protocol describes the use of midigels and minigels. PMID- 18428447 TI - Standard laboratory equipment. AB - This appendix unit lists standard pieces of equipment for the modern neuroscience laboratory. More specialized equipment is also itemized in the materials list of each protocol throughout the series. In the individual materials lists no attempt has been made to list all items required for each procedure, but only those items that might not be readily available in the laboratory or that require special preparation. PMID- 18428448 TI - Preparation of an iodinated radioligand. AB - Radioactive tracers are useful for receptor measurement and for radioimmunoassays because of the ease with which radioactive ligands are detected. Typical receptor measurement methods include whole tissue receptor assays, autoradiography, and cell and membrane binding assays. The isotopes most commonly used in receptor measurement studies are tritium, carbon-14, and iodine-125, with the choice of isotope most often based on the specific activity required and the sensitivity and type of detection system available. Its high specific activity and easy detection make I-125 ideal for labeling large peptides and proteins. While the short half-life (60 days) of iodine requires frequent replacement with freshly iodinated compounds, this is also an advantage, as unused portions of radioligand can be held for decay and disposed of as nonradioactive waste. This unit describes the iodination of proteins or peptides using the 125I-labeled Bolton Hunter reagent, a convenient, easy-to-use, nonoxidizing labeling reagent. PMID- 18428449 TI - High precision stereotaxic surgery in mice. AB - This unit provides protocols for cannulation and site-specific central microinjection of mice using a recently developed high-precision stereotaxic frame. The construction of cannulae, wire plugs and injection needles are also described. PMID- 18428450 TI - Receptor binding techniques. AB - This overview first discusses issues relating to the selection of radioligand for receptor binding assays, including the isotopic label and considerations pertaining to the pharmacological and chemical profile of the ligand. This is followed by a section on characterization of ligand-binding assays, starting with tissue preparation methods, followed by detection of specific binding, determination of incubation and washing conditions and a discussion of saturation and competition assay formats. Quantification of the assay results can be accomplished by autoradiography or film densitometry. Finally, methods and considerations for analysis of the resulting data are presented. PMID- 18428451 TI - Transneuronal circuit analysis with pseudorabies viruses. AB - Over the past decade there has been a dramatic increase in the use of viruses as transneuronal tracers of neuronal circuitry. The method exploits the propensity of neurotropic viruses to invade neurons and then produce infectious progeny that cross synapses to infect other neurons within a circuit. The protocols and commentaries included in this unit focus upon the use of the swine alpha herpesvirus known as pseudorabies virus (PRV) for polysynaptic analysis. Here, the aspects of experimental design that have the greatest import for successful use of viruses in circuit definition are presented. Accordingly, the protocols included in this unit can be applied in concert with methods in which the use of classical tract tracers has been detailed. A procedure for retrograde infection of CNS circuits in the rat CNS by peripheral injection of virus is detailed, while transneuronal analysis by intracerebral injection is also described. A variant of these procedures, transneuronal analysis with multiple recombinant strains, is also described along with methods for growing and titering viral stocks, and procedures for single and dual immunohistochemical localization of viral antigens in fixed brain tissue. PMID- 18428452 TI - Metabolic mapping. AB - This unit describes a high-resolution [(3)H]2-deoxyglucose (2DG)/immunohistochemistry double labeling protocol which enables simultaneous visualization of metabolic and immunohistochemical markers (e.g., neurotransmitter-specific epitopes) in the same tissue section. This approach generates single-cell metabolic measures from large (many thousands) samples of neurons, and allows the assignment of putative neurotransmitter types to large samples of functionally assayed single neurons. Thus, it provides a unique opportunity to assay the metabolic activities of histochemically identified neural elements throughout an entire pathway. [(3)H]2DG injection and perfusion fixation of hamsters or mice is described first, followed by immunohistochemical staining of brain slices for a monoclonal antibody against glutamic acid decarboxylase-6 (GAD-6). Included are the steps for dipping and developing slides for autoradiography. An alternate protocol gives details of staining for cytochrome oxidase (CO). PMID- 18428453 TI - Targeted toxins. AB - Molecular neurosurgery can be used to make selective neural lesions by targeting cytotoxins to specific populations of neurons based on their common expression of a particular surface molecule. The targeted toxins employed in this unit consist of a targeting moiety (vector) and an effector moiety (cytotoxin). In all cases discussed in this unit, the cytotoxic moiety is an enzyme that catalytically inactivates the large ribosomal subunit, irreversibly inhibiting protein synthesis and resulting in cell death. These toxins appear to kill in an all-or none fashion, indicating that one molecule of free cytotoxin in the cytoplasm of a cell is sufficient to kill the cell. Three general molecular neurosurgery protocols are presented in this unit. The first describes suicide transport, which refers to the use of targeted toxins to make anatomically restricted lesions based on retrograde axonal transport of the toxin. The second involves immunolesioning and uses anti-neuronal immunotoxins to make type-selective and anatomically restricted lesions. The final protocol uses neuropeptide-toxin conjugates to selectively destroy neurons expressing the receptor for the specific neuropeptide. PMID- 18428454 TI - Cellular compartment analysis of temporal activity by fluorescence in situ hybridization (catFISH). AB - This protocol describes a method (cellular compartment analysis of temporal activity by fluorescent in situ hybridization or catFISH) that uses fluorescent in situ hybridization to immediate-early gene RNAs and confocal microscopy to identify neuronal populations activated at two distinct times. The combination of cellular and temporal resolution makes catFISH a valuable tool for investigating the dynamic interactions of neuronal populations associated with different behaviors or cognitive challenges. PMID- 18428455 TI - Detection of DNA damage in tissue sections by in situ nick translation. AB - The technique of in situ nick translation (ISNT) is used to detect DNA strand breaks in tissue sections at the cellular level with great sensitivity. In fact, ISNT can be used to detect DNA damage in a single cell, which is particularly useful to assess programmed cell death during development. One crucial advantage of ISNT is the anatomical resolution that permits a detailed topographical analysis of DNA damage. This can be useful to identify cells that are more vulnerable to an experimental insult. Furthermore, cells with DNA damage can be identified morphologically with this method. This can be of interest to determine whether cells that exhibit DNA damage already exhibit clear features of dying cells or are still relatively intact morphologically. This can be useful to identify the mode of cell death involved. This unit provides a protocol that describes tissue preparation, in situ nick translation and emulsion autoradiography. PMID- 18428456 TI - Theory and application of fluorescence microscopy. AB - The growing importance in biology and especially in neurobiology of fluorescence microscopy is due to (1) the extraordinary development of new fluorescent molecular probes and (2) the development of improved low light level imaging systems and confocal microscopy techniques. This overview covers fluorescent molecular probes, filters and filter sets, multiband filters and multidye fluorescence, light sources, microscope objectives, image resolution and the point-spread function, and general steps for immunolabeling. PMID- 18428457 TI - Basic confocal microscopy. AB - Confocal microscopy produces sharp images of structures within relatively thick specimens (up to several hundred microns). It is particularly useful for examining fluorescent specimens. This overview intended to provide background and practical tips needed to get started with confocal microscopy. It begins with a description of the basis of optical sectioning, then discusses various types of confocal microscopes, and concludes with practical guidelines for sample preparation and optimizing image acquisition parameters. PMID- 18428458 TI - Imaging nervous system activity. AB - Optical imaging methods rely upon visualization of three types of signals: (1) intrinsic optical signals, including light scattering and reflectance, birefringence, and spectroscopic changes of intrinsic molecules, such as NADH or oxyhemoglobin; (2) changes in fluorescence or absorbance of voltage-sensitive membrane dyes; and (3) changes in fluorescence or absorbance of calcium-sensitive indicator dyes. Of these, the most widely used approach is fluorescent microscopy of calcium-sensitive dyes. This unit describes protocols for the use of calcium sensitive dyes and voltage-dependent dyes for studies of neuronal activity in culture, tissue slices, and en-bloc preparations of the central nervous system. PMID- 18428459 TI - Culture of neuroepithelial stem cells. AB - In vitro culture of multipotent neural precursors allows researchers to study mechanisms regulating such processes as proliferation and lineage commitment within the developing central nervous system (CNS). The protocols presented in this unit describe the isolation and maintenance of stem cells from both fetal and adult rodent neural tissue. A procedure for treating tissue culture dishes with poly-L-ornithine and fibronectin, which is necessary before the dishes are used for neuroepithelial stem cell culture, is also included. PMID- 18428460 TI - Calcium phosphate transfection of DNA into neurons in primary culture. AB - Calcium phosphate/DNA coprecipitation is a widely used method for the introduction of foreign DNA into cells. DNA and calcium phosphate are allowed to form a precipitate that is then added to cells in culture. The cells internalize the DNA, leading to the expression of the transfected genes in the cell. Despite the simplicity of this method, it has not been used very often for primary neurons because of its potential to cause neuronal toxicity. However, low toxicity and reasonably high transfection efficiency (0.5% to 5%) can be achieved by optimization of the transfection parameter, combined in some cases with the use of inhibitors of neuronal activity. This unit describes a very easy and inexpensive method for neuronal gene delivery that can be used with standard eukaryotic expression vectors for the gene of interest. PMID- 18428461 TI - Purification of oligodendrocytes and their progenitors using immunomagnetic separation and Percoll gradient centrifugation. AB - In this unit, two techniques are described for the purification of oligodendrocytes and their progenitors from the developing mammalian central nervous system (CNS). The first method utilizes the technique of immunomagnetic separation to selectively isolate oligodendrocytes and their progenitor cells from the optic nerve of prenatal and early postnatal rats. This technique takes advantage of the surface antigens expressed on these cells. A paramagnetic bead is attached to the cells via an antibody bridge. Target cells that are coupled to magnetic beads can then be separated from a heterogeneous cell population using a magnetic field. The second method for isolating oligodendrocytes uses Percoll gradient centrifugation to separate oligodendrocytes from a heterogeneous cell population by virtue of their cell density and allows the direct isolation of oligodendrocytes from animals aged postnatal day 4 (P-4) to adult. This method is particularly useful for assessing physiological systems present in development that may be lost as a result of growing purified neonatal cells in vitro in the absence of neuronal influence. PMID- 18428462 TI - Isolation and generation of oligodendrocytes by immunopanning. AB - This unit presents a procedure for the purification of oligodendrocyte progenitor cells, their expansion in vitro, and differentiation of these cells to yield oligodendrocyte cultures. A variation is also presented, detailing the direct isolation of differentiated oligodendrocytes from postnatal brain. The purification of the target cell population is achieved by exploiting the differential binding of cells to tissue culture dishes coated with an antibody directed against a specific cell-surface antigen. Cells expressing this surface antigen are retained on the dish and are thereby separated from the remaining cell population. PMID- 18428463 TI - Generation and utilization of phosphorylation state-specific antibodies to investigate signaling pathways. AB - Phosphorylation state-specific antibodies can be of great use, for example, in studying individual steps within a given signal transduction pathway. This unit presents a general approach to the generation and purification of phosphorylation state-specific antibodies. In addition to their ability to detect phosphorylation at a particular key site, these antibodies are often more sensitive for biochemical studies. Besides their application in immunoblotting procedures, activation state-specific antibodies can be used as immunohistochemical reagents. Thus, critical changes in phosphorylation can be monitored as described on an individual cell basis or in fixed tissue sections. Such antibodies can be used to address fundamental questions about signal transduction pathways during physiologic events that cannot be resolved by more conventional methodologies. PMID- 18428464 TI - Neuronal transfection using particle-mediated gene transfer. AB - This unit describes the use of particle-mediated gene transfer (also known as biolistics) for the transfection of neuronal cell lines and brain slices. Like nuclear microinjection of DNA, biolistics results in the direct introduction of DNA into the nucleus; it is perhaps for this reason that biolistics works as well in mitotic cells as in postmitotic cells such as skeletal muscle, skin, liver, and neurons. The basic principle of biolistics is to accelerate micron-sized gold particles coated with DNA towards target cells or tissue. Cells penetrated by these particles have a high likelihood of being transfected by the DNA thus introduced. The motive force for particle acceleration can come from a variety of sources, the most widely used is described in this unit and is a supersonic shock wave generated by the rupture of a kapton membrane induced by high-pressure helium. Another option included in this unit is to propel the gold particles by gas jet entrainment. PMID- 18428465 TI - Gene expression in transgenic mice using neural promoters. AB - In the first part of this unit, major considerations for the analysis of neural promoters in transgenic mice are discussed. Detailed protocols on the production of transgenic mice are not described in this unit. Advantages and disadvantages of the transgenic approach for analysis of neural cis-acting elements are also presented. The concept of transient transgenic mice is then introduced; this method compensates for some disadvantages associated with the conventional transgenic approach. Finally, major factors influencing the efficiency of transgenic mouse production are discussed. The second part of the unit presents detailed information on a variety of neural-specific cis-acting elements that have been characterized by a transgenic approach. This information is useful both as a guide for carrying out the analysis of cis-acting elements and as a reference for selection of promoter/enhancer elements for designing an appropriate transgenic construct. PMID- 18428466 TI - Isolation and purification of primary Schwann cells. AB - Schwann cells are the glial cells of the peripheral nervous system (PNS) which both myelinate and provide trophic support for their associated axons. Their functions are critical for proper development, homeostasis, and regeneration of the PNS. Schwann cells can be isolated and expanded in culture. Such a culture can be used as a source of myelinating glia in culture or as a source of Schwann cells for various cell and molecular biological experiments. The proliferative ability of these cells is exploited in the isolation procedures presented here. PMID- 18428467 TI - Culture of substantia nigra neurons. AB - Primary cultures of nigral tissue are widely used as a model system to assay effects of trophic and toxic agents on dopaminergic neurons. Cultured dopaminergic neurons have been successfully transplanted in animals and led to behavioral improvement in animal models of Parkinson's disease. Cell cultures have also been used to study the development of substantia nigra, allowing investigators to identify early inductive events important for nigral development and to study dopaminergic differentiation and target innervation. This unit provides simple and reliable culture protocols for these applications. The first approach presented is the preparation of dissociated nigral cell cultures, the later steps of which can be used as a simple and efficient assay for testing growth factors. A second approach is the preparation of free-floating roller tube cultures, which may be used as a tool for neural transplantation and to study more complex developmental events. A third approach is the production of organotypic cultures using chicken plasma as a matrix. Organotypic cultures can maintain the in vivo cytoarchitecture of a host region in vitro. PMID- 18428468 TI - Isolation and purification of primary oligodendrocyte precursors. AB - Oligodendrocytes are the myelinating cells of the central nervous system, and are avidly studied by investigators interested in neural cell lineages, myelination, multiple sclerosis, and oligodendroglioma. This unit presents a strategy for manipulating the division and differentiation of oligodendrocyte progenitors by isolating O-2A progenitors from optic nerve and then purifying and expanding them by culture in a defined medium supplemented with platelet-derived growth factor (PDGF) plus basic fibroblast growth factor (bFGF). Cells are then allowed to differentiate through removal of the growth factors. A protocol is also provided for monitoring the cell type composition of the cultures by immunohistochemistry. In addition to making it possible to generate large numbers of cells, this approach allows access to intermediate differentiation stages. Cells can thus be studied as they differentiate into oligodendrocytes, making the actual process of differentiation open to examination. PMID- 18428469 TI - Isolation and purification of primary rodent astrocytes. AB - Astrocytes are a major cell type in the mammalian central nervous system (CNS). The ability to obtain virtually pure populations of these cells makes it possible to study their function as isolated cells or in mixed populations where they support the growth and survival of surrounding neurons. Unlike other mature CNS cells, mature astrocytes maintain the lifelong ability to reenter the cell cycle. The first isolation procedure described in this unit takes advantage of the proliferative ability of these cells, as does the second, except that no antibody or complement treatment is required. A procedure for detecting glial fibrillary acidic protein (GFAP), which is present in most astrocytes in vivo and virtually all astrocytes in vitro and is a useful marker for assessing the purity of cultures, is also presented. PMID- 18428470 TI - Differentiation of embryonic stem cells. AB - Embryonic stem (ES) cells are pluripotent cells whose developmental state is equivalent to cells of the inner cell mass in the blastocyst-stage embryo. This unit presents a method of producing differentiated ES cells in which the cells are first aggregated on a less adhesive surface to form embryoid bodies (EBs). EBs are allowed to attach to a permissive substrate and then differentiate into neural cells in a serum-free medium. The resulting cell population can be maintained for several passages in serum-free medium in the presence of basic fibroblast growth factor (bFGF) or allowed to differentiate into postmitotic neurons by plating onto a glial monolayer followed by culture with serum. Also included in the unit are procedures for preparing feeder cells from embryonic fibroblasts and for preparing glial monolayer cultures, both for use in the main ES production protocol. PMID- 18428471 TI - Immortalizing central nervous system cells. AB - This unit presents methods for isolating clonal, neural-derived cell lines. One approach for isolating such neural cell lines involves a replication-deficient retrovirus encoding a specific oncogene and a selectable marker which are used to infect dissociated CNS cells dissected at a developmental stage at which the cell population of interest has not undergone its terminal mitotic division. Also presented is a method for cloning by limiting dilution, which may be necessary to obtain a pure population of cells. Following growth under appropriate selection conditions, clones are isolated and tested for their ability to differentiate with the desired phenotypic properties. A method is also provided for coating tissue culture dishes, which is necessary for successful culture of CNS neurons. PMID- 18428473 TI - Homology cloning of cDNA or genomic DNA. AB - The principle of cloning utilizes hybridization of single-stranded DNA probes to denatured DNA fixed on membranes to detect the identical DNA molecules present in a large population of diverse DNAs. In homology or low-stringency cloning, hybridization of oligonucleotide, cDNA, or genomic DNA probes to denatured DNA from plasmid or bacteriophage libraries is carried out under low-stringency conditions that promote hybridization in the presence of sequence mismatch (i.e., mispaired nucleotides). The procedures used are identical to those for screening libraries at high stringency, with the exception that hybridization and washing conditions are altered to permit hybridization with mismatched sequence; salt concentration is increased and the hybridization and wash temperatures are decreased. This unit describes conditions that have been used to clone and identify novel genes and cDNA clones using low-stringency hybridization of known probes to membranes that contain libraries of bacterial or bacteriophage DNA. PMID- 18428472 TI - Identification of neural programmed cell death through the detection of DNA fragmentation in situ and by PCR. AB - A universal feature in the development of multicellular organisms is a physiological form of cell death called programmed cell death (PCD). A subset of PCD is apoptosis, which is defined by characteristic morphological changes and genomic DNA fragmentation producing what are referred to as nucleosomal ladders. To understand how PCD operates in a developing tissue or in a tissue following an experimental procedure, dying cells must be identified in relation to their surviving neighbors. One way to accomplish this is to visualize fragmented DNA in situ, in conjunction with gel electrophoresis of isolated DNA to visualize the nucleosomal ladders associated with apoptosis. Two approaches are presented in this unit: in situ end-labeling plus (ISEL+), a technique to identify dying cells in tissue sections or cell cultures of central nervous system (CNS) tissue (optimized for embryonic samples); and the use of ligation-mediated polymerase chain reaction (LMPCR) to identify nucleosomal ladders from intact tissues. Also included are procedures for preparing thymocyte cell cultures for use as controls in the ISEL+ procedure and for isolating genomic DNA for LMPCR. PMID- 18428474 TI - Oligonucleotide-directed mutagenesis without phenotypic selection. AB - A DNA sequence can be specifically altered by synthesizing the desired sequence change within an oligonucleotide, and then converting this into a biologically active circular DNA strand by using the oligonucleotide to prime in vitro synthesis on a single-stranded circular template. This unit presents a protocol which uses a DNA template containing a small number of uracil residues in place of thymine. Use of the uracil-containing template allows rapid and efficient recovery of mutants; in principle this same template can be applied to most of the other mutagenesis protocols in use. The length of the oligonucleotide primer is highly variable and depends on the nature of the change being made. PMID- 18428475 TI - Directed mutagenesis using the polymerase chain reaction. AB - The polymerase chain reaction (PCR) is most often used for the enzymatic amplification and direct sequencing of small quantities of nucleic acids. This technology can also be used as a quick and efficient method for introducing any desired sequence change into the DNA of interest. This unit contains two basic protocols for introducing base changes into specific DNA sequences. The first describes the incorporation of a restriction site and the second details the generation of specific point mutations. An alternate protocol describes generating point mutations by sequential PCR steps. Although the general procedure is the same in all three protocols, there are differences in the design of the synthetic oligonucleotide primers and in the subsequent cloning and analyses of the amplified fragments. PMID- 18428476 TI - Overview of gene delivery into cells using HSV-1-based vectors. AB - This overview describes the considerations involved in the preparation and use of herpes simplex virus type 1 (HSV-1) as a vector for gene transfer into neurons. Strategies for gene delivery into neurons, either to study the molecular biology of brain function or for gene therapy, must utilize vectors that persist stably in postmitotic cells and that can be targeted both spatially and temporally in the nervous system in vivo. This unit describes the biology of HSV-1 along with a discussion covering development of amplicon and genomic HSV-1 vectors. Advantages and disadvantages of current HSV-1 vectors are presented, and HSV-1 vectors are compared with other vectors for gene transfer into neurons. PMID- 18428477 TI - Generation of high-titer defective HSV-1 vectors. AB - There are two types of replication-deficient herpes simplex virus type 1 (HSV-1) vectors: those in which the foreign DNA of interest is cloned into the viral genome itself, and those that are comprised of a plasmid (amplicon) carrying minimal HSV-1 sequences that allow it to be packaged into virus particles with the aid of a helper virus. This unit describes the generation of helper virus stocks. Preparation of recombinant amplicon vector particles by transfection of amplicon and superinfection of helper virus into cells, and harvesting of packaged particles, is also delineated. Thorough characterization of the viral vector stock involves measuring (1) the helper virus plaque-forming units per ml (pfu/ml) on 2-2 cells, (2) the wild-type HSV-1 pfu/ml on Vero cells, and (3) the amplicon stock infectious vector units per ml (ivu/ml) on PC12 cells. Support protocols detail methods for determining titers of helper virus and wild-type virus by plaque assay, and of amplicon stocks by vector assay. PMID- 18428478 TI - The substituted-cysteine accessibility method (SCAM) to elucidate membrane protein structure. AB - The substituted-cysteine accessibility method (SCAM) provides an approach to identifying the residues in the membrane-spanning segments that line a channel, transporter, or binding-site crevice. SCAM can also be used to determine differences in the structures of the membrane-spanning segments in different functional states of the proteins, to map electrostatic potential in the membrane spanning domains, and to size a channel or binding-site crevice. The protocol in this unit describes the use of SCAM to map the binding-site crevice of a G protein coupled receptor (GPCR) which binds ligand within the transmembrane portion of the receptor. PMID- 18428479 TI - Overview of the baculovirus expression system. AB - Baculoviruses have emerged as a popular system for overproducing recombinant proteins in eukaryotic cells. This unit gives an overview of the baculovirus expression system, including discussion of the baculovirus life cycle, and post translational modifications that occur in insect cells. In addition, the steps for overproducing proteins in the baculovirus systems are described along with recommendations for choosing an appropriate baculovirus vector and DNA, and reagents and equipment necessary for implementing the whole overexpression system. PMID- 18428480 TI - Protein expression in the baculovirus system. AB - Insect cell-recombinant baculovirus co-cultures offer a protein production system that complements microbial systems by providing recombinant proteins in soluble form and with most post-translational modifications. Moreover, the large size of the viral genome enables cloning of large segments of DNA and consequent expression of complex protein aggregates. This unit describes methods associated with the large-scale production of recombinant proteins in the baculovirus expression system. A method for large-scale production of viral stocks is described and methods for titration of virus are provided (a plaque assay and an end-point assay). Once viral stocks have been prepared and titered, a protocol for testing the virus in small-scale cultures is provided to determine the kinetics of expression, which allows evaluation of various cell culture and infection conditions aimed at developing optimal levels of protein production (e.g., comparisons of different host cell lines, media, and environmental parameters). Support protocols provide instructions for preparing culture samples for protein analysis by SDS-PAGE and discuss analytical methods for monitoring nutrient levels in cell culture fluids. Once optimal process parameters are identified, protocols describe production of the target protein on a large scale in fermentors using either regular batch production in bioreactors or a fed-batch procedure of production in perfusion cultures. Techniques for harvesting cultures from bioreactors are also provided. PMID- 18428481 TI - PCR cloning of neural gene products. AB - Of the many proteins that are known to be involved in neuronal signaling, one family of gene products, collectively referred to as the G protein-coupled receptors (GPCRs), has received considerable attention. Within the transmembrane domains of GPCRs are clusters of amino acids that tend to be conserved among receptors that bind related ligands. Polymerase chain reaction (PCR)-based approaches to cloning novel GPCRs typically begin with the identification of these well-conserved amino acid motifs, which are then back-translated into degenerate oligonucleotide primers. These pools of degenerate oligonucleotides are the most important variables in PCR cloning of GPCRs. Although GPCRs are used as the focus of this unit, the strategies and techniques described are applicable to the cloning of a wide variety of neuronal gene products. In the first procedure in this unit, either total or poly(A)(+) purified RNA is reverse transcribed into first-strand cDNA. In subsequent steps the cDNA product serves as the template for synthesis and amplification of target receptor sequences by PCR primed with degenerate oligodeoxynucleotides. The product is ready to be cloned and screened as described. Guidelines for database searching are provided to help identify the cloned gene from the known sequence. Typically, only a portion of the receptor coding region is cloned by the above approach. Rapid amplification of cDNA ends (RACE) or anchored PCR is described in this unit and is used to obtain a full-length cDNA amenable for expression studies. PMID- 18428482 TI - Calcium flux assay in Xenopus oocytes. AB - Many G protein-coupled receptors of interest to neuroscientists induce transient increases in [Ca(2+)](i), which can be used as a convenient measure of receptor activation in a variety of applications. This unit describes a simple calcium flux assay applied to Xenopus oocytes. PMID- 18428483 TI - Gene transfer into neural cells in vitro using adenoviral vectors. AB - Adenoviruses (Ads) have become a very attractive and versatile vector system for delivering genes into brain cells in vitro and in vivo. One of the main attractions of Ads is that they can mediate gene transfer into post-mitotic cells, i.e. neurons. Ads are easy to grow and manipulate, stable, and their biology is very well understood. This unit is designed to help newcomers into the field, to design, prepare and grow replication-defective recombinant adenovirus vectors with the aim of transferring genes into neurons and glial cells in primary culture. It provides step-by-step methods describing the preparation of brain cell cultures, their infection using recombinant adenovirus vectors and also the assessment of transgene expression using a variety of techniques including fluorescence immunocytochemistry and fluorescence activated cell sorting (FACS) analysis. The methods described will be useful to scientists wishing to enter the adenovirus field to construct adenovirus vectors to be used for gene transfer into neural cells. PMID- 18428485 TI - Overview of nucleic acid arrays. AB - Nucleic acid array technology refers to the use and fabrication of arrays containing thousands of nucleic acid samples bound to solid substrates such as glass microscope slides or silicon wafers. Because the physical area occupied by each sample is usually 50 to 200 micrometers in diameter, it is possible to assay nucleic acid samples representing entire genomes, ranging in size from 3,000 to 32,000 genes, on a single slide. Microarrays are useful for analyzing gene expression patterns, genotyping and genetic mapping, comparative genomic hybridization, polysome analysis, and DNA-protein interactions. This overview describes the technology and applications, and provides valuable web site listings for obtaining additional information. PMID- 18428484 TI - Gene transfer into rat brain using adenoviral vectors. AB - Recombinant adenovirus vectors are attractive vehicles to deliver genes into the brain for the purposes of neurobiological research and for gene therapy of neurological diseases. This unit provides a comprehensive set of protocols for adenovirus vector-mediated gene transfer to the brain, including introduction of the vector into the brain by stereotaxic injection and preparation and processing of brain tissue for the evaluation of gene transfer. The potential side-effects of administering adenovirus vectors to the brain are discussed in detail. The unit also provides protocols for evaluating these side-effects (e.g., demyelination, inflammation, vector-mediated cytotoxicity, etc.). Finally, critical parameters for obtaining optimal gene transfer with minimum side-effects are presented. PMID- 18428486 TI - Preparation of mRNA for expression monitoring. AB - The ability to construct comprehensive gene expression profiles comprising hundreds to thousands of genes whose RNA levels are monitored simultaneously represents an exciting new capability in molecular biology. This is accomplished by hybridizing mRNA, which has been quantitatively amplified and labeled with biotin, to DNA chips that display thousands of nucleotides complementary to the mRNAs of interest. In this unit, rationale for starting with poly(A(+)) versus total RNA is discussed, and strategies for choosing oligonucleotides for chip design is presented. Protocols on RNA amplification and labeling, and purifying and quantifying the cDNA and in vitro transcription products are included. PMID- 18428487 TI - Expression cloning of neural genes using Xenopus laevis oocytes. AB - Expression cloning requires a representative cDNA or genomic DNA library and a host organism in which the cloned genes can be transcribed and/or translated. It likewise requires a method to detect the expressed protein using, for example, the inherent biological activity of the gene or antibodies specific for the gene product. Most successful expression cloning strategies have employed cDNA libraries constructed in plasmid or bacteriophage lambda vectors and Xenopus oocytes or cultured mammalian cells as hosts. This unit presents several protocols designed for expression cloning paradigms that rely on electrophysiological recordings from Xenopus laevis oocytes. PMID- 18428488 TI - Overview of neural gene expression. AB - This overview describes issues that must be considered before attempting to express neural cDNAs in mammalian cells, including the choice of expression vector and cell type. Considerations for introducing recombinant vectors into cells are discussed along with a comparison of achieving stable or transient expression. Finally, the appropriate promoter is crucial and must be chosen to fit the design of the expression system. PMID- 18428489 TI - Selection of transfected mammalian cells. AB - Analysis of gene function frequently requires the formation of mammalian cell lines that contain the studied gene in a stably integrated form. Approximately one in 10(4) cells in a transfection will stably integrate DNA (the efficiency can vary depending on the cell type). Therefore, a dominant, selectable marker is used to permit isolation of stable transfectants. In the first part of this unit, the procedure for determining selection conditions and the resulting stable transfection is presented and the most commonly used selectable markers are discussed. The second protocol includes conditions for thirteen markers commonly used for selection of mammalian cells. A third protocols describes selection of transfected cells from the total population soon after transfection with plasmids that express both the gene of interest and a selection tag. Optimization of transfection conditions can be facilitated by a simple staining assay detailed in a support protocol. PMID- 18428490 TI - Transient expression of proteins using COS cells. AB - This unit describes the use of COS cells to efficiently produce a desired protein in a short period of time. These cells express high levels of the SV40 large tumor (T) antigen, which is necessary to initiate viral DNA replication at the SV40 origin. Each COS cell transfected with DNA encoding a cell-surface antigen (in the appropriate vector) or cytoplasmic protein will express several thousand to several hundred thousand copies of the protein 72 hr posttransfection. If the transfected DNA encodes a secreted protein, up to 10 mg of protein can be recovered from the supernatant of the transfected COS cells 1 week posttransfection. COS cell transient expression systems have also been used to screen cDNA libraries, to isolate cDNAs encoding cell-surface proteins, secreted proteins, and DNA binding proteins, and to test protein expression vectors rapidly prior to the preparation of stable cell lines. PMID- 18428491 TI - cDNA expression cloning in mammalian cells. AB - This unit contains protocols for expression cloning in mammalian cells. Either calcium phosphate- or liposome-mediated transfection of mammalian cells, or virus infection and liposome-mediated transfection are used to screen pools derived from a cDNA library. cDNA pools are prepared for cloning from library-transformed E. coli grown in liquid culture medium or on antibiotic-containing selection plates. Results of screening assays for expression can be detected using autoradiography of dishes of cultured cells to identify clones, direct visualization of radiolabeled cells on emulsion-coated and developed chamber slides, detection and quantification of gene activity by a functional (transport) assay with scintillation counting, or detection using a filter-based assay for binding of radioligand to membranes or whole cells. The most critical step of any cDNA cloning project is the establishment of the screening protocol. Therefore, the bioassay for the gene product must be established prior to executing any of these protocols, including construction of the cDNA library. PMID- 18428492 TI - Mutagenesis approaches for elucidation of protein structure-function relationships. AB - Several mutagenesis strategies have been used to allow the identification of specific receptor-ligand interactions to support the docking of ligands within various receptor models. The strengths and limitations of each strategy are discussed in this overview. Large-scale mapping is described using deletion and chimeric receptors. Fine tuning with single residue replacements is also covered along with two-dimensional approaches. PMID- 18428493 TI - Metal-chelate affinity chromatography. AB - Recombinant proteins engineered to have six consecutive histidine residues on either the amino or carboxy terminus can be purified using a resin containing nickel ions (Ni(2+)) that have been immobilized by covalently attached nitrilotriacetic acid (NTA). This technique is know as metal-chelate affinity chromatography and can be performed using either native or denatured protein. This unit presents protocols for expression of histidine-tail fusion proteins and their purification in either native or denatured form (along with procedures for renaturation by either dialysis or solid-phase renaturation). Also provided are procedures for analysis of the purified produce and regeneration of the NTA resin. PMID- 18428494 TI - Immunoaffinity chromatography. AB - This technique involves elution of a single protein from an immunoaffinity column after prior elution of nonspecifically adsorbed proteins. Antibodies are coupled to Sepharose and the cell lysate is passed over the column. After washing, the specific antigen is eluted from the column using one of a number of methods: brief exposure to either high or low pH, or using the detergent octyl beta-D glucoside (easily removable by dialysis). All these methods are described in this unit along with a procedure for covalently linking an antibody to Sepharose using the cyanogen bromide activation method. PMID- 18428495 TI - The use of phage display in neurobiology. AB - Phage display is a technique that involves the coupling of phenotype to genotype in a selectable format. It has been extensively used in molecular biology to study protein-protein interactions, receptor and antibody binding sites, and immune responses; to modify protein properties; and to select antibodies against a wide range of different antigens. In the format most often used, a polypeptide is displayed on the surface of a filamentous phage by genetic fusion to one of the coat proteins, creating a chimeric coat protein. As the gene encoding the chimeric coat protein is packaged within the phage, selection of the phage on the basis of the binding properties of the polypeptide displayed on the surface simultaneously results in the isolation of the gene encoding the polypeptide. This unit describes the background of the technique and illustrates how it has been applied to a number of different problems, each of which has its neurobiological counterparts. PMID- 18428496 TI - Fluorescent in situ transcription in cells and tissues. AB - This method assesses cellular mRNA transcripts in tissue sections and cell cultures using unique short anti-sense primers directed against sequences in particular protein(s). The unlabeled synthetic cDNA oligonucleotide primers are extended complementary to a sense mRNA transcript using reverse transcriptase and labeled through incorporation of a fluorescent-labeled dUTP nucleotide base. The new cDNA will be synthesized upstream from the point of primer hybridization, and has a specific activity of fluorescent labeling dependent upon the length of the template mRNA from the primer location to the 5'-terminus. This procedure provides rapid detection of low abundance mRNA messages that can be related to other cellular protein components, labeled experimentally with alternative fluorochromes. PMID- 18428497 TI - Detection of protein phosphorylation in tissues and cells. AB - Phosphorylation is one of the principal regulatory mechanisms in the nervous system. Several different procedures used to characterize the phosphorylation state of neuronal proteins are described in this unit, including analysis of phosphorylation in situ, phosphoamino acid analysis, and phosphopeptide map analysis. In addition, there is a protocol describing in vitro phosphorylation of fusion proteins. These methods are often combined to provide a comprehensive evaluation of the phosphorylation state of a particular protein. PMID- 18428498 TI - Use of the A. victoria green fluorescent protein to study protein dynamics in vivo. AB - Fluorescent molecules serve as valuable tools for the detection of numerous biochemical phenomena and have been employed for protein localization, quantitation of gene expression, detection of nucleic acids, cell sorting and determination of chemical concentrations. However, the use of such techniques generally requires significant nonphysiological perturbations to the biological system being studied; therefore, they are not always appropriate for the observation of dynamic phenomena. Green fluorescent protein (GFP), cloned from jellyfish, has been used to overcome many of these problems. It is a small, extremely stable fluorescent protein that has been successfully expressed and detected in a wide variety of organisms, both in intact form and fused to other proteins. This overview unit describes the use of this proteinaceous fluorophore for in vivo observation of cellular phenomena. PMID- 18428499 TI - Green fluorescent protein in the study of neuronal signaling pathways. AB - In recent years, techniques have been established for transiently co-transfecting cells with cDNA of the jellyfish green fluorescent protein (GFP), a reporter gene that encodes a non-toxic marker. This approach can be applied to primary neurons where it has become especially useful for the study of neuronal second messenger pathways. This unit describes procedures for transfecting neurons in primary culture: transfection with GFP DNA, including co-transfecting with separate GFP and gene-of-interest constructs, transfecting with a single construct containing the gene of interest fused to a GFP gene, and transfecting with a single construct containing separate gene-of-interest and GFP cassettes. Also included is a method for the rapid, large-scale preparation of a nearly homogeneous population of neurons from rat cerebellum. The Commentary provides several examples of how this approach can be applied to specific biological questions on neuronal signaling pathways. PMID- 18428500 TI - Analysis of RNA by northern and slot-blot hybridization. AB - Specific sequences in RNA preparations can be detected by blotting and hybridization analysis using techniques very similar to those originally developed for DNA. Fractionated RNA is transferred from an agarose gel to a membrane support (northern blotting), while unfractionated RNA is immobilized by slot or dot blotting. The resulting blots are studied by hybridization analysis with labeled DNA or RNA probes. Included in this unit are detailed procedures for RNA denaturation, blotting and hybridization. Also described is a method for stripped hybridization probes from blots so the blots can be re-hybridized with a different probe. PMID- 18428501 TI - Differential display of mRNA by PCR. AB - This unit outlines the polymerase chain reaction (PCR)-based technique of mRNA differential display, which identifies genes that are differentially expressed between cells or tissues. The approach described here is a modification of the original method, referred to as single base-anchored differential display. The basic protocol describes the actual differential display PCR reaction along with details of the identification, reamplification, and cloning of candidate differentially expressed genes. The support protocol provides instructions on removing contaminating genomic DNA from the RNA samples and reverse transcribing the purified RNA to produce the cDNA used in the subsequent PCR reactions. PMID- 18428502 TI - Analysis of mRNA populations from single live and fixed cells of the central nervous system. AB - This unit presents a method for the amplification of poly(A)(+) mRNA extracted from the cytoplasm of a single cell. After cDNA is synthesized from the mRNA, it is made double stranded, denatured, and reverse transcribed to yield antisense RNA (aRNA). Another round of amplification results in a relatively large amount of aRNAs in essentially the same proportion as in the starting mRNA population. RNA amplification protocols can be used for many purposes, including generation of disease expression profiles, making of cDNA libraries, and generation of diagnostics and therapeutics for disease. An alternate protocol is used to amplify RNAs from single neurons in fixed tissue specimens. Support protocols gives instructions for reverse northern analysis, which allows analysis of the presence or absence and relative levels of mRNA expression in selected cells, and a convenient method to assess the RNA content in fixed tissue sections using the fluorescent dye acridine orange (which binds single-stranded nucleic acids). PMID- 18428503 TI - Reducing gene expression in the brain via antisense methods. AB - This unit presents protocols that employ antisense oligodeoxynucleotides to reduce expression of target proteins in the brain. These oligonucleotides are generally designed to inhibit synthesis of a specific protein by hybridization to its mRNA. Because oligonucleotides show very poor penetration into the central nervous system (CNS) after systemic administration, they are either injected into the cerebrospinal fluid (CSF) or infused directly into the brain parenchyma. In this unit, the procedure most commonly used for delivering oligonucleotides continuously into CSF is outlined. In addition, a procedure is described for continuous infusion of oligonucleotides into a specific brain region, using the substantia nigra as an example. PMID- 18428504 TI - Production of polyclonal antisera. AB - Antibodies are serum immunoglobulins with binding specificity for particular antigens, and polyclonal antibodies are particularly valuable for immunoprecipitation and immunoblotting. In this unit, the production of polyclonal antisera specific for protein antigens in rabbits, rats, mice, and hamsters is described. A support protocol is included for preparing serum from blood. PMID- 18428505 TI - Production of antipeptide antisera. AB - This unit describes methods used for the chemical coupling of synthetic peptides to carrier proteins, required for the preparation of peptide immunogens. The carrier protein described here is keyhole limpet hemocyanin (KLH) because it is the one most commonly used. However, other proteins may be used in place of KLH, including bovine serum albumin (BSA) and ovalbumin. Coupling may be accomplished as described with MBS, which requires a Cys residue in the peptide, or with glutaraldehyde, EDCI, or BDB. Also included are an assay for detecting free sulfhydryl (SH) groups, a means of calculating coupling efficiency, an immunization schedule, an indirect ELISA, and a method for preparing a peptide affinity column. The final methodology described is the multiple antigen (MAP) system. PMID- 18428506 TI - Epitope tagging of recombinant proteins. AB - Epitope tagging is a method of expressing proteins whereby an epitope for a specific monoclonal antibody is fused to a target protein using recombinant DNA techniques. The fusion gene is cloned into an appropriate expression vector for the experimental cell type and host cells are transfected. The fusion protein can then be detected and/or purified using a monoclonal antibody specific for the epitope tag. This unit presents protocols for detection and purification of proteins tagged with a particular epitope, the FLAG tag, although the same general approach can be applied to other epitope tags. The protocols in this unit employ the anti-FLAG M2 antibody to detect and purify FLAG-tagged proteins. The methods presented are immunoprecipitation of FLAG fusion proteins from cells using an anti-FLAG M2 affinity gel, detection of FLAG fusion proteins by western blotting, and purification of FLAG fusion proteins by anti-FLAG M2 affinity chromatography. PMID- 18428507 TI - Overview of membrane protein solubilization. AB - The solubilization of membrane proteins (i.e., extraction of the protein from the membrane in a disperse state in aqueous solution) in a stable state that retains the native activity of the membrane-bound form is more an empirical art than a protocol-driven science. Although a large number of molecules have been invented that display detergent properties, selection of the proper solubilizing agent is largely a matter of trial and error that depends on the protein of interest and the particular goals of the investigator. The purpose of this overview is to describe the methodology for finding the optimal conditions for detergent solubilization and to relate the selection of a specific detergent to explicit experimental goals. PMID- 18428508 TI - The electrophysiology setup. AB - This unit describes components and considerations that are common to all electrophysiology setups dedicated to measurement of electrical activity in cells. First, the basic concerns of setting up the equipment in a functional and accessible manner are described. Then two typical setups are briefly described: one for in vitro extracellular recording, the other for single-channel patch clamping. An electrophysiological setup has four main requirements: (1) environment: the means of keeping the preparation healthy; (2) optics: the means of visualizing the preparation; (3) mechanics: the means of stably positioning the microelectrode; and (4) electronics: the means of amplifying and recording the signal. This unit addresses primarily the mechanics and electronics of the electrophysiological laboratory setup. PMID- 18428509 TI - Recording and analyzing synaptic currents and synaptic potentials. AB - Intracellular recording of synaptic currents (PSCs) under voltage clamp conditions provides the most accurate and direct means for measuring the earliest effects of neurotransmitters. With this tool, combined with pharmacological or ionic manipulations, one can obtain information about the type of transmitter used at a synapse, the dynamics of transmitter-receptor interactions, the types and numbers of receptors activated, the effects of drugs on transmission, functional neural circuitry, and indications about the mechanisms of synaptic plasticity. Each synaptic current or potential is a reflection of many experimental variables: the ionic composition of the solutions, the temperature, the presence of pharmacological agents, the rate of synaptic stimulation, the history of stimulation, the variables of the recording system, as well as other factors unique to each preparation. Correct analysis of data requires all these parameters be considered. Both stimulus-evoked and spontaneous synaptic events are covered in this unit since conclusions about synaptic and drug mechanisms are strongest when based upon recording of both types of activity. This unit outlines basic considerations for recording PSCs and PSPs in addition to guidelines for data analysis. PMID- 18428510 TI - Preparation and maintenance of organotypic slice cultures of CNS tissue. AB - Organotypic slice cultures are the in vitro method of choice for applications requiring long-term survival of the preparation and a high degree of cellular differentiation and organization resembling that of the original tissue. Long term survival is achieved by culturing slices at the air/liquid interface, either by continuously rotating the preparation (roller-tube cultures) or by culturing them on semiporous membranes (stationary interface cultures). Both culture techniques yield nerve cells which are highly differentiated in terms of their morphological and physiological characteristics. Because most of these cultures are prepared from 1-week-old postnatal animals, in which the cellular and tissue organization is already relatively advanced, the original cytoarchitecture is often remarkably well maintained. Moreover, the presence of a full complement of glial and nerve cells is thought to provide a microenvironment facilitating differentiation of neurons. Slice culture also offers unique advantages for recording from pairs of cells, as a consequence of the high degree of connectivity between nerve cells. Recently, new applications have emerged such as the cultivation of slices from knock-out animals with limited postnatal survival time or alteration of gene expression by viral vectors. PMID- 18428511 TI - Voltage clamp recordings from Xenopus oocytes. AB - Xenopus oocytes serve as a standard heterologous expression system for the study of cloned ion channels. The large size of these cells allows for relatively easy expression and recording of activity of exogenous ion channels (together with neurotransmitter receptors and/or various regulatory proteins) using the whole cell two-electrode voltage clamp (TEVC) technique, as well as standard single channel patch clamp recordings. Although usually advantageous, the cell size also dictates certain limits on the accuracy of recordings and requires specific modifications of recording methods. However, combining the advantages of the system with available recording methods enables the use of Xenopus oocytes for sophisticated multidisciplinary studies of ion channels. PMID- 18428512 TI - Synaptic plasticity in the hippocampal slice preparation. AB - Synaptic plasticity is the process by which the brain alters the strength of its synaptic connections, a fundamental function of the brain that enables individuals to learn from experience. The study of synaptic plasticity often involves the application of standard in vitro electrophysiological techniques to hippocampal slice preparations. This unit discusses many of the special considerations that are applicable for the optimal study of synaptic plasticity in this system. Most of these principles also apply to the study of synaptic plasticity in other brain slice preparations. PMID- 18428513 TI - Principles of signal conditioning. AB - It is rare for biological, physiological, chemical, electrical, or physical signals to be measured in the appropriate format for recording and interpretation. Usually, a signal must be conditioned to optimize it for both of these functions. This overview describes the fundamentals of signal filtering, how to prepare signals for A/D conversion, signal averaging to increase the signal-to-noise ratio, line frequency pickup (hum), peak-to-peak and rms noise measurements, blanking, audio monitoring, testing of electrodes and the common mode rejection ratio. PMID- 18428514 TI - Preparation of hippocampal brain slices. AB - This unit presents a procedure for the preparation of acute mammalian hippocampal slices for electrophysiological recording. Although this protocol should not be taken as the only means of making brain slices, it is a widely-used typical procedure. It is simple and straightforward. It can be used on a variety of mammalian species, though it is discussed here for use in rats. PMID- 18428515 TI - Acute isolation of neurons from the mature mammalian central nervous system. AB - The acute dissociation procedure provides a simple means of isolating neurons from the mature mammalian central nervous system. The method was primarily devised to isolate neurons for patch-clamp electrophysiology. It may also prove useful for single-cell PCR, immunocytochemistry, sorting of fluorescently labeled cells, or long-term tissue culture of mature neurons. Dissociation is brought about by a combination of proteolysis and an ionic environment that encourages breakdown of the tissue. The method allows the isolation of neurons free of glial ensheathments in as little as 45 min after the sacrifice of the animal. Neurons so isolated lose fine dendritic branches, although the structure proximal to the cell body is often maintained, allowing identification of the morphological type of the neuron. The preparation has the following advantages: (1) the neurons are fully differentiated; (2) the cells are electronically compact, which improves the fidelity of the voltage clamp; (3) the cells are removed from the influence of surrounding cells; and (4) neurons can be isolated from small, circumscribed loci within the adult central nervous system. PMID- 18428516 TI - Whole-cell voltage clamp recording. AB - This unit describes the use of whole-cell voltage clamping to study voltage-gated channels. Stepwise changes in voltage produced by this technique cause channels to interconvert between different states, and these transitions are monitored as changes in membrane current. The time course of this redistribution of states contains a great deal of information about the mechanism of channel gating. Furthermore, the voltage clamp can be used to activate different populations of channels selectively. In this way, a specific channel targeted by biological or pharmacological manipulations can often be identified and studied in detail. This technique is also readily adapted to the study of ligand-gated channels, synaptic potentials, and exocytosis. PMID- 18428517 TI - Patch-pipet recording in brain slices. AB - Patch-clamp recording in brain slices provides a powerful approach for investigating the intrinsic electrical properties of neurons and glia and analyzing synaptic interactions between neurons. There are two main methods presently in use to obtain patch-clamp recordings from neuronal or glial cell bodies and their processes in brain slices, both described in this unit. In the blind technique described in this unit, the approach to a cell is performed blindly and recordings are obtained relying purely on electrical cues. Alternatively, the movement of the recording pipet through the brain slice and its placement onto the cell membrane can be performed as described under visual control; this method is usually performed with the aid of differential interference contrast (DIC) optics and is referred to as the DIC technique. Increased resolution of cells and their processes can be obtained by combining DIC optics with infrared illumination and video microscopy techniques. Using either of these methods it is possible to reliably apply the full power of the patch-clamp technique to the study of neurons and glial cells in brain slices. PMID- 18428518 TI - Single-channel recording. AB - This unit provides detailed descriptions for the steps of patch excision, data acquisition, and data analysis, and elaborates upon the relevant issues discussed in other units from Chapter 3. It includes discussions of the instrumentation for single-channel recording and the key concepts necessary for the interpretation of single-channel data. PMID- 18428519 TI - Rapid solution application methods. AB - Solution changes to deliver solutes of different compositions are required in virtually every cellular electrophysiological experiment. Also, in many neurobiology experiments, it is necessary to make rapid step changes in the concentration of a test compound in order to outpace receptor desensitization or to mimic the brief lifetime of a fast synaptic response. The goal of this unit is to aid the investigator in choosing the rapid solution application method that is most appropriate for the experimental situation and to describe the methods for fabricating two relatively simple devices for making rapid changes between different solutions. PMID- 18428521 TI - Measurement of chloride movement in neuronal preparations. AB - In this unit, protocols are described for biochemical and optical techniques that have been used by investigators to measure ligand-gated chloride movement in vesicular structures called synaptoneurosomes (also referred to as microsacs), in cultured neurons, and in the acute brain slice. These techniques can be applied to other ions as well. The measurement of uptake and efflux of radioisotopic chloride in synaptoneurosomes is used to study the responses of gamma aminobutyric acid (GABA) receptors, which are coupled to chloride channels. Similar chloride flux assays for primary neuronal cultures are also presented. Alternatively, the efflux of chloride from synaptoneurosomes and primary neuronal cultures can be studied using fluorescent dyes and photometry. Finally, the measurement of chloride uptake can be studied in individual neurons in brain slices using fluorescent dyes and optical imaging by nonconfocal and confocal microscopy. Several support protocols are provided as well, outlining the preparation of synaptoneurosomes from specific brain regions, and the preparation, loading, and calibration of chloride-sensitive fluorescent dyes. PMID- 18428520 TI - Overview of microdialysis. AB - The technique of microdialysis enables the monitoring of neurotransmitters and other molecules in the extracellular environment. This method has undergone several modifications and is now widely used for sampling and quantitating neurotransmitters, neuropeptides, and hormones in the brain and periphery. This unit describes the principles of conventional and quantitative microdialysis as well as strategies in designing a dialysis experiment. It establishes the groundwork for the basic techniques of preparation, conduct, and analysis of dialysis experiments in rodents and subhuman primates. Although the methods described are those used for monitoring CNS function, they can be easily applied with minor modification to other organ systems. PMID- 18428522 TI - Measurement of cation movement in primary cultures using fluorescent dyes. AB - Ca(2+), Na(+), K(+), and Mg(2+) have a central role in neuronal excitability. The concentration of these cations in the cytoplasm of neurons (generically termed [ion(+)]i) provides a marker of the excitation state of the neurons, and may also illuminate the activity of specific signaling mechanisms that involve Ca(2+)- or Mg(2+)-activated enzymes. The measurement of [ion(+)]i in cultured neurons is achieved with the use of an ion-sensitive fluorescent dye in combination with equipment designed to quantitatively measure fluorescence. Specificity is obtained by choosing dyes with the appropriate selectivity for the ion of interest. Measurements of steady state ion concentrations can be made, as well as measurements of the net difference between ion movement into the cytoplasm (in response to a stimulus) and the physiological buffering of that ion. The procedures in this unit for loading and recording from dyes are broadly similar for each ion when ratiometric dyes are used as described, and can readily be modified for use with single-wavelength dyes. Support protocols are provided for calibration of individual dyes, which can be more problematic. PMID- 18428523 TI - Measurement of second messengers in signal transduction: cAMP and inositol phosphates. AB - cAMP acts as an intracellular mediator of hormone action and the importance of accurate quantitative determination of cAMP levels in cells and tissues is widely recognized. The most utilized procedures for the determination of adenylate cyclase activity in membranes are described here for measuring the conversion of [alpha-(32)P]ATP into [(32)P]cAMP after a two-step chromatographic separation. Also critical in signal transduction is phosphoinositide turnover, which is linked to receptor activation resulting from changes in cytosolic calcium concentrations. Phosphoinositide turnover can be measured as described in this unit by labeling phospholipid pools with [(3)H]-inositol and then analyzing for tritiated inositol phosphates. PMID- 18428524 TI - Measurement of NO and NO synthase. AB - Nitric oxide (NO) is a key biosignaling molecule produced in both peripheral tissues and the central nervous system by a family of enzymes known as nitric oxide synthases (NOSs). NOSs convert L-arginine to stoichiometric quantities of NO and L-citrulline using molecular oxygen and NADPH as cofactors. Techniques for measurement of NO and NOS activity are essential to demonstrate the role of NO and NO-derived species in biological systems. This unit describes two methods for detection of NO: a direct method employing chemiluminescent detection and one based on quantification of the stable oxidation products with detection using the Griess reagent. Additionally, NOS activity can be quantified by measuring the conversion of radiolabeled L-arginine to radiolabeled L-citrulline. PMID- 18428525 TI - Measurement of nitric oxide in single cells and tissue using a porphyrinic microsensor. AB - This unit describes the preparation and applications of porphyrinic sensors for quantitative measurement of nitric oxide (NO) in single cells and in tissues. The determination of NO is based on the electrochemical oxidation of NO on a carbon fiber electrode covered with a thin layer of a conducting polymeric metalloporphyrin catalyst, overlaid with another thin film of Nafion, a cation exchange material. The electric current generated during NO oxidation on the surface of the polymeric porphyrin is linearly proportional to the concentration of NO, so this current is used as an analytical signal which can be measured in either the amperometric or the voltammetric mode. Both methods provide a quantitative signal. This unit describes the electrochemical setup for measurement of NO in single cells and tissue. Support protocols describe porphyrin synthesis, sensor preparation, and sensor calibration. PMID- 18428526 TI - Peripheral type benzodiazepine binding sites as a tool for the detection and quantification of CNS injury. AB - The concentration of peripheral type benzodiazepine binding sites (PTBS) in the brain parenchyma is greatly increased following brain lesions, reflecting the glial reaction and/or presence of hematogenous cells. Thus, PTBS density is a sensitive and reliable marker of brain injury in a large number of experimental models (ischemia, trauma, excitotoxic lesions, brain tumors) and equivalent human neuropathological conditions. PTBS density can be measured using specific radioligands and a conventional binding technique, or by quantitative autoradiography in tissue sections. PMID- 18428527 TI - Measurement of oxygen radicals and lipid peroxidation in neural tissues. AB - An important role for oxygen radical-mediated neuronal damage has been implicated in a number of acute and chronic neurodegenerative disorders. Particular interest has centered upon oxygen radical-induced, iron-catalyzed lipid peroxidation (LP) as the principal mechanism of the neuronal injury associated with oxygen radicals. Thus, there has been a growing interest in methods for monitoring increased oxygen radical levels as an index of oxidative stress as well as markers of LP-associated oxidative injury in a number of in vitro and in vivo model systems. This unit provides a detailed description of the salicylate trapping method for the measurement of the most highly reactive oxygen radical, the hydroxyl radical, as well as several direct or indirect methods for assessment of cellular LP in either cell cultures or in in vivo models. PMID- 18428528 TI - Assessment of cell viability in primary neuronal cultures. AB - Four commonly used methods for the assessment of neuronal (or glial) cell viability are described in this unit. The MTT assay is sensitive to the function of labile mitochondrial enzymes, which typically lose activity early in the progression towards death. The lactate dehydrogenase (LDH) assay measures the appearance of this cytosolic enzyme in the bathing medium, providing a measure of plasma membrane integrity. Loss of plasma membrane integrity is also the basis of the trypan blue dye assay and the propidium iodide assay. Trypan blue staining is assessed by cell counts; propidium iodide labeling can be assessed either by cell counts, typically in conjunction with fluorescein diacetate counterstaining to identify intact cells containing adequate levels of functional esterases, or with a fluorescence plate reader. PMID- 18428529 TI - In vivo measurement of blood-brain barrier permeability. AB - This unit describes various protocols for the in vivo quantitation of drug permeability across the rodent blood - brain barrier. Methods for the measurement of drug influx or efflux are described, and support protocols are provided for determining intravascular capillary volume and cerebral perfusion flow. An in situ perfusion technique is also provided for assessing whether transport of a test compound occurs by carrier-mediated or saturable transport. PMID- 18428530 TI - Microdialysis in rodents. AB - Microdialysis is an established sampling technique for the in vivo measurement of a variety of substances in both blood and tissue. These include neurotransmitters and neuropeptides, enzymes, and electrolytes, as well as various hormones and pharmaceutical agents. More recently, microdialysis has been used to infuse exogenous as well as endogenous substances into the brain and spinal cord. In microdialysis, a semipermeable dialysis membrane is introduced into the fluid or tissue compartment to be sampled and perfused with physiological fluid. As a result of passive diffusion, molecules migrate across the membrane along their concentration gradient. Molecules found in high concentrations within the tissue compartment migrate across the membrane into the dialysis tubing where they can be collected for subsequent quantification, whereas molecules found in high concentrations within the membrane diffuse outward into the surrounding tissue compartment. This unit describes construction of concentric and side-by-side microdialysis probes, which differ with respect to size and inflow/outflow tube compositions, as well as a modification of a horizontal probe. This unit also covers methods for surgically implanting microdialysis probes in rats and mice and for conducting microdialysis in vitro and in vivo in rodents. Finally, procedures are provided for carrying out quantitative microdialysis techniques. PMID- 18428531 TI - Microdialysis in nonhuman primates. AB - This unit presents a technique that allows for routine repeated microdialysis experiments in the monkey using a sedated preparation or, with further minor modification, in the awake behaving animal. Protocols are provided for construction and in vitro calibration of dialysis probes, obtaining magnetic resonance imaging (MRI) scans of the monkey brain, surgically attaching a guide holder to the skull, and performing postoperative MRI scanning, in vivo dialysis sample collection, anatomical verification of the probes, and neurochemical analysis. An alternate protocol describes microdialysis in awake, behaving monkeys; this requires substantial preparatory work in training the monkeys to sit quietly in a restraining chair and/or to perform a series of behavioral tasks. PMID- 18428532 TI - Saturation assays of radioligand binding to receptors and their allosteric modulatory sites. AB - The protocols in this unit describe methods for measuring radioligand binding to sites on the GABA-A receptor complex. This system was chosen because of the fundamental role of GABA-A receptors in central nervous system function and the presence of multiple binding sites on one supramolecular complex. However, these basic techniques can be used to analyze ligand binding to a wide variety of intracellular and extracellular sites associated with ion channels, transporters, or G proteins with relatively simple modifications of the protocols (e.g., tissue preparation, buffers, assay termination techniques, incubation times, and temperatures). A saturation assay is presented to examine ligand binding to the benzodiazepine site on the GABA-A receptor, while a second protocol uses a similar technique to examine the binding site for GABA in this complex. PMID- 18428533 TI - Saturation analysis of ligand binding using a centrifugation procedure. AB - The use of filtration assays is often unsuitable for radioligands with rapid dissociation rates. Rapid dissociation may result in the loss of significant amounts of bound ligand during the separation of bound from free ligand associated with filtration. The primary advantage of the centrifugation assay for characterizing the binding of a radioligand is that the bound ligand is separated from free ligand without initiating significant ligand dissociation from its receptor. Nonetheless, the small volume of supernatant that remains trapped within the membrane pellet (even after a superficial rinsing of the pellet surface) often results in an increased nonspecific binding compared to filtration techniques. The unit presents a protocol for the binding of [(3)H]glycine to the glycine recognition site of the NMDA receptor complex. This assay is commonly employed because of the current lack of high-affinity agonist ligands that interact with this recognition site. The use of other radioligands may require different tissue preparations, buffer systems, and assay conditions, but the basic steps involving sedimentation of the pellet, removal of supernatant, superficial washing of the pellet surface, and solubilization of the pellet will remain the same. PMID- 18428534 TI - Ligand characterization using microphysiometry. AB - This unit describes the use of a Cytosensor microphysiometer for functional characterization of an agonist and antagonist to a G protein-coupled receptor, the muscarinic M1 receptor. Concentration-response profiles are used to calculate values for the EC50 of the response of cells to the agonist and the pA2 value for the antagonist. Support protocols describe optimization of two aspects of this procedure: the duration of ligand exposure at a given concentration and the length of recovery time between the administration of two different concentrations of ligand to minimize the impact of desensitization. The Cytosensor microphysiometer allows the measurement of receptor activation in both adherent cells, such as the M1WT3 cells used here or in suspension cultures. PMID- 18428535 TI - Uptake and release of neurotransmitters. AB - The availability of clonal cell lines for norepinephrine, dopamine, and serotonin transporters allows the characterization of drug interactions with transporter recognition sites using radioligands, as well as the characterization of drug effects on selective transporter-mediated uptake and release of substrate. In addition to clonal cell lines, synaptosomes prepared from specific brain regions can be used to conduct these studies without interference by endogenous transporters or binding proteins that are present in other tissues. This unit presents protocols for uptake and release of tritiated substrates using intact cells (either detached or in suspension) or synaptosomes. An HPLC procedure for electrochemical detection of nonradiolabeled substrates is also provided. Time dependent release can also be measured in assays involving real-time sampling. PMID- 18428536 TI - Rodent models of depression: forced swimming and tail suspension behavioral despair tests in rats and mice. AB - Rodents forced to swim in a narrow space from which there is no escape will, after an initial period of vigorous activity, adopt a characteristic immobile posture, making only those movements necessary to keep their heads above the water. It was hypothesized that immobility reflected the animals' having learned that escape was impossible and their having given up hope. Immobility was therefore given the name "behavioral despair". Immobility was subsequently found to be reduced by a wide range of clinically active antidepressant drugs. This simple behavioral procedure has since become a useful test for screening novel antidepressants in rats and is presented in this unit. An equivalent procedure in the mouse is also described along with a "dry" version of the test where immobility is induced simply by suspending the mouse by the tail. PMID- 18428537 TI - Rodent models of depression: learned helplessness using a triadic design in rats. AB - Certain types of human depression are precipitated by stressful life events, and vulnerable individuals experiencing these stressors may develop clinical depression. Understanding the neurobiology of stress vulnerability (depression) as well as stress resiliency (coping) is critical for guiding the development of novel pharmacotherapeutic agents for stress-related disorders such as depression in humans. The use of a triadic design (escapable shock, yoked-inescapable shock and restrained control) allows the investigator to examine the various sequella of stress exposure, while manipulating and quantifying the impact of psychological dynamics of stress such as active behavioral coping (i.e., stress control). Both escape and yoked subjects are exposed to the identical amount, intensity, pattern and duration of stress. The critical distinction between these two groups is that the escape group has the opportunity to terminate the shock stress by turning a wheel at the front of a chamber, while wheel-turning for the yoked subject is of no consequence. Any difference observed between the escape and yoked subjects is a result of the effects of coping, rather than stress exposure per se. The restrained group is included to control for the effects of handling. Any differences between this group and the escape and yoked subjects reflects the impact of stress per se. PMID- 18428538 TI - Rodent models of depression: learned helplessness induced in mice. AB - Uncontrollable stressors induce a variety of behavioral disturbances that are in many ways reminiscent of the symptoms that characterize clinical depression. These deficits are evident across a range of species, including mice. Given the increasing focus on genetic techniques involving mice to identify the mechanisms subserving these behavioral disturbances (e.g., recombinant, knockout, and transgenic strains), it is of particular interest to provide a detailed description of the method to induce behavioral deficits in response to uncontrollable stressors. This unit describes the procedure used to assess the effects of controllable and uncontrollable shock on subsequent shock escape performance in mice using an escape-delay procedure. PMID- 18428539 TI - Fear-potentiated startle in rats. AB - Fear-potentiated startle measures the increase in the startle reflex elicited by a sudden noise in the presence of a cue that has previously been paired with footshock. It involves an initial training session in which a cue, such as a light, is paired with footshock, and then a later test session in which startle is elicited in the presence or absence of the cue. It thus involves associative learning, memory for that association measured during the retention test, and conditioned fear in the presence of the cue. Drugs that interfere with learning, memory, or fear and anxiety can alter fear-potentiated startle depending on when they are given with respect to training and testing. PMID- 18428540 TI - Motor coordination and balance in rodents. AB - Measurement of motor coordination and balance can be used not only to assess the effect of drugs or other experimental manipulations on mice and rats, but also to characterize the motor phenotype of transgenic or knock-out animals. Three well established and widely used protocols for measuring motor coordination and balance in mice and rats (rotarod, beam walking and footprint analysis) are described in this unit. The tests can be used equally well for rats and mice, and have been used both for the phenotypic characterization of transgenic mice and for evaluating the effects of lesions and aging in rats. The protocols are described in the primary context of testing mice, but modifications of the test apparatus or variations in the test parameters for assessment of rats are noted. PMID- 18428541 TI - Latent inhibition. AB - Latent inhibition (LI) refers to the deleterious effects of nonreinforced stimulus preexposure on its subsequent conditioning. LI is considered to index an organism's ability to ignore inconsequential stimuli and as such provides a useful tool to measure attentional processes as well as their disruption and enhancement by various pharmacological, physiological and behavioral manipulations. During the last decade, LI has received increasing interest as an animal model of schizophrenia and of antipsychotic drug action. This unit describes the measurement of LI in the conditioned emotional response and two-way active avoidance procedures, including parametric variations which allow the demonstration of the disruption and the potentiation of this phenomenon. PMID- 18428542 TI - Sexual and reproductive behaviors. AB - The procedures described in this unit include testing procedures for male and female reproductive behaviors, gonadectomy, and hormonal treatments appropriate for inducing male and female reproductive behaviors. Because reproductive behaviors are social behaviors, and therefore require the presence of stimulus animals, the protocols in this unit also provide information on the preparation of stimulus animals. The protocols are written for use with laboratory rats, although a discussion of issues related to species differences in the study of reproductive behaviors is included. PMID- 18428543 TI - Assessment of spatial memory using the T maze. AB - This unit describes the use of the T maze to assess spatial memory, and takes into account the alternating behavior of rats in searching for food. The task is based on the premise that animals have evolved an optimal strategy to explore their environment and obtain food with a minimum amount of effort. The T maze has been most extensively used to investigate specific aspects of spatial working memory, which is operationally defined as information that is only useful to a rat during the current experience with the task. A modification of the maze allows for the assessment of reference memory, defined as information that is useful across all exposures to the task (i.e., on any day of testing). Finally, in the absence of food-deprivation, a simple T maze can be used as described to assess spontaneous alternation. PMID- 18428544 TI - Overview of methodological approaches to the study of ingestive behavior. AB - Some examples of procedures used by feeding researchers are discussed in this unit and include ablation of neural function; inhibition of behaviors by selective neurotoxins and antisense oligonucleotides; staining of sensory and motor mechanisms; electrical stimulation of the brain; local injection and microdialysis of nutrients, neurotransmitters, and drugs; autoradiography and in situ hybridization of neurotransmitters with their receptors; electrophysiological techniques for multi- and single-unit recording of cells in the hypothalamus; and gene technology using inbred strains of genetically obese mice. PMID- 18428545 TI - Basic measures of food intake. AB - This unit describes a method for measuring the consumption of a pellet diet during a 30-min testing session conducted during the late portion of the day. The procedure generates reliable and stable baseline measures of feeding, and is appropriate for assessing experimental manipulations that either enhance (e.g., injecting a peptide into the brain) or suppress feeding (e.g., systemic injections of amphetamine). PMID- 18428546 TI - Sham feeding in rats with chronic, reversible gastric fistulas. AB - Sham feeding occurs when ingested food is prevented from accumulating in the stomach and small intestine by surgical intervention. Sham feeding permits the investigation of the orosensory controls of meal size in the absence of postingestive controls. This unit details preoperative preparation, surgery, postoperative care and maintenance, and test procedures for the simplest and most widely used method, which involves implantation of a gastric cannula in a rat to provide a chronic, reversible gastric fistula. PMID- 18428547 TI - Conditioned flavor preferences: evaluating postingestive reinforcement by nutrients. AB - The acceptance of food and the expression of preferences for particular foods have both innate and learned components. To determine the mechanism(s) responsible for the acquisition of learned preferences and the enhancement of inborn taste preferences, it is important to separate the component stimuli: the oral flavor cues and the postingestive nutrient cues that are associated with the flavors. Unambiguous analysis of postingestive reinforcement of a nutrient requires post-oral presentation of the nutrient during or after oral intake of the flavor stimulus. This protocol concentrates on controlling stimulus presentation in associative conditioning, pairing a flavor with positive postingestive effects. PMID- 18428548 TI - Measurement of startle response, prepulse inhibition, and habituation. AB - The startle response is comprised of a constellation of reflexes elicited by sudden, relatively intense stimuli. The startle reflex is useful for studying fundamental properties of nervous function ranging from neurophysiological and anatomical relationships within the pons and reticular formation to forebrain regulation of complex behavioral states and cognitive processes. This unit presents protocols for measurement of the startle response in rats to acoustic stimuli, along with modifications encompassing an experimental manipulation such as drug treatment and measures of habituation of startle; a method for measuring prepulse inhibition (PPI) of startle; and specialized rat handling and calming techniques that supplement the startle protocols. PMID- 18428549 TI - Measurement of rodent stereotyped behavior. AB - This unit presents a quantitative, observational method for the assessment of rodent stereotyped behavior which consists of motor responses that are repetitive, invariant, and seemingly without purpose or goal. The most classic behavioral pattern that is characteristic of stereotypy is that elicited by high doses of stimulants, such as cocaine and amphetamine, in rats, although it can also occur in response to other drugs or neurotoxic treatments affecting the basal ganglia. An observational time-sampling procedure is described in which animals are observed and rated by an experimenter, who is blind to treatment, at regular time points over the course of a behavioral testing period. The frequency of different behaviors is measured by scoring the presence or absence of a given behavior during predetermined time bins. The apparatus and test procedures are described, and a comprehensive list of commonly observed behaviors that may appear as stereotyped is provided. In addition to being ideally suited to the measurement of stereotypy, the protocol can be adapted to sampling many forms of spontaneous behaviors, including locomotion, rearing, grooming, eating, and drinking. Samples of behavioral checklists and scoring sheets are also provided. PMID- 18428550 TI - Dural inflammation model of migraine pain. AB - The cause of migraine pain is controversial. One recently proposed theory is that migraine pain may originate from inflammation of the meninges, particularly the dural membranes that surround the brain. This theory proposes that, during a migraine, there is an idiopathic activation of trigeminal sensory afferents, resulting in nociceptive transmission to the CNS as well as the release of pro inflammatory substances in the periphery, particularly the dura. Dural inflammation is thought to lower the nociceptive threshold of dural afferents and facilitate nociceptive transmission to the central nervous system. In the procedure described in this unit, trigeminal sensory afferents are activated by electrically stimulating the trigeminal ganglion. This stimulation causes trigeminal peripheral sensory afferents to depolarize, inflammatory substances to be released from these afferents, and dural inflammation to appear. Dural inflammation is quantified by measuring plasma protein extravasation. The basic protocol describes this model in rats, and the alternate protocol describes the analogous procedure in guinea pigs. PMID- 18428551 TI - Preclinical models of hepatic encephalopathy. AB - Hepatic encephalopathy is a multifactorial neuropsychiatric syndrome accompanying acute or chronic liver failure. Techniques for developing animal models of hepatic encephalopathy associated with acute or chronic liver failure, or vascular shunting are illustrated. In addition, the behavioral and biochemical characteristics of these models are described. PMID- 18428552 TI - Preclinical models of Parkinson's disease. AB - Parkinson's disease is a neurodegenerative disease in which pigmented midbrain neurons progressively die producing a dopamine (DA) deficit in the striatum which manifests as an akinetic movement disorder. Experimentally induced striatal DA depletion in animals is a valid model of parkinsonism. The capacity of certain substances to damage catecholaminergic neurones has been used for a long time to produce DA deficiency in animals. This unit focuses on methods for inducing parkinsonism using the neurotoxins MPTP and 6-hydroxy dopamine and methods for evaluating the animals. Other models are briefly reviewed. PMID- 18428553 TI - Rodent models of global cerebral ischemia. AB - Brain damage after stroke and head injury remains a huge clinical problem. In stroke, the initial cause of the damage is a blockage in a blood vessel (often the middle cerebral artery) and this sets off several pathways that ultimately lead to cell death. Recent studies have demonstrated that several new mechanisms are involved in neuronal death and this has led to an increase in research into novel molecules that might prevent brain damage or improve recuperation post stroke. There are several models of global cerebral ischemia. Two of the most widely-used models are discussed in detail in UNIT 9.5, the gerbil bilateral carotid artery occlusion (BCAO) model and rat 4-vessel occlusion (4-VO) model. Additionally, several models of focal cerebral ischemia have been developed to mimic the effects of human stroke. The rationale behind the use of animal models, the various types of models and advantage and disadvantages of each model are presented. PMID- 18428554 TI - Rodent models of focal cerebral ischemia. AB - This unit presents models that are both used to study ischemic mechanisms and to test for neuroprotective agents or agents that enhance recovery from stroke. The Tamura model is one of the best characterized focal ischemia models in which the middle cerebral artery is occluded by electrocoagulation. Also described is the intraluminal monofilament model, the spontaneously hypertensive rat (SHR), and the newer endothelin-1 model. The rationale behind the use of animal models, the various types of models and advantage and disadvantages of each model are presented. PMID- 18428555 TI - Experimental autoimmune encephalomyelitis (EAE). AB - The procedures described in this unit utilize murine models exclusively since murine EAE often results in a relapsing/remitting disease, similar to the early phase of most MS patients. EAE in the Lewis rat is a monophasic illness in which animals experience a single episode of paralysis from which most recover completely. This unit presents two methods for inducing EAE in mice: active induction and adoptive transfer. PMID- 18428556 TI - Animal health assurance. AB - A program to assure the health of laboratory animals and to provide a stable environment for their maintenance is necessary to reduce unwanted variables or complicating factors in experimentation. To assure animal health throughout a research study, the animals involved should be procured in good health, monitored for continued health status, and protected from pathogenic organisms through testing of any biological materials. This unit addresses all three of these health management practices. Essential in support of these practices is a diagnostic laboratory capable of testing for rodent and rabbit pathogens. PMID- 18428557 TI - Electronic imaging in neuroscience. AB - This unit is intended to aid the neuroscientist in understanding the basics of image detectors and selecting a suitable camera for various neuroscience research applications. A procedure is described for evaluating cameras in the laboratory. PMID- 18428558 TI - Loading neurons with dextran-conjugated calcium indicators in intact nervous tissue. AB - This unit describes methods for filling populations of neurons and their processes, including presynaptic terminals, with dextran-conjugated calcium indicators in central nervous tissue of mammals and lower vertebrates. Techniques for filling neurons in vivo for subsequent analysis either in vivo or in brain slices or en bloc preparations are described. These methods are also suitable for staining neurons in acute and organotypic brain slices. PMID- 18428559 TI - cDNA library construction from single cells. AB - Analysis of gene expression or comparing gene expression patterns between cells and/or tissues facilitates the identification of molecules activated by a cellular program or by a particular physiological or pharmacological treatment. This unit presents a protocol for studying these molecules at the level of a single cell. PMID- 18428560 TI - Gene expression analysis using cDNA microarrays. AB - This unit focuses on cDNA microarrays, which are being increasingly applied to large-scale gene expression studies in the nervous system, concentrating on the steps of probe preparation and microarray hybridization. Four protocols for the attachment of fluorophores to cDNA (labeling) and cDNA hybridization onto microarrays are presented: direct labeling using either Klenow fragment or reverse transcriptase, and two signal amplification techniques represented by the tyramide signal amplification method (TSA), and detection and labeling of cDNA using PCR amplification. Basic issues in experimental design, sources of variability and potential pitfalls in microarray experiments are described. Anticipated results and follow up of microarray experiments, including confirmation of the results and troubleshooting are also discussed. PMID- 18428561 TI - Using phage display in neurobiology. AB - In this unit some of the basic protocols involved in the manipulation of phage display libraries are described, including the rescue and amplification of such libraries, selection and screening from them and testing of derived clones. PMID- 18428562 TI - Amperometry and cyclic voltammetry with carbon fiber microelectrodes at single cells. AB - Amperometry and cyclic voltammetry are two electrochemical techniques that enable the detection of electroactive neurotransmitters that are released from single cells. These techniques have provided the first chemical view of the events that occur during exocytosis. This unit describes the isolation of several cell types known for their exocytotic properties, the fabrication and calibration of carbon fiber microelectrodes, as well as some of the equipment and software requirements for obtaining electrochemically generated data. PMID- 18428563 TI - Preparation and maintenance of organotypic cultures for multi-electrode array recordings. AB - Recording from neuronal cultures with multi-electrode arrays (MEAs) provides a powerful tool for studying neuronal activity with many neurons simultaneously in vitro. This unit describes the detailed steps necessary for growing organotypic cultures on MEAs and the typical neuronal activity that is obtained with this methodology. PMID- 18428564 TI - Methods for sample preparation for direct immunoassay measurement of analytes in tissue homogenates: ELISA assay of amyloid beta-peptides. AB - Use of low abundance analytes in whole tissue homogenates has been realized with the development of assays in which a specific analyte is captured and detected using immunological reagents. One of the many advantages of analyte immunoassay in crude homogenates is its relative simplicity, allowing high throughput analysis of samples. In this unit, some major key determinants in sample and standard preparation and handling are described that have been shown to improve the performance and reliability of these assay systems. The ELISA assay of amyloid peptides from brain tissue is described as an example, since the protocols for this analysis exemplify many of the techniques and problems that are encountered in the development of new assays. PMID- 18428565 TI - Analyzing radioligand binding data. AB - Radioligand binding experiments are easy to perform, and provide useful data in many fields. They can be used to study receptor regulation, discover new drugs by screening for compounds that compete with high affinity for radioligand binding to a particular receptor, investigate receptor localization in different organs or regions using autoradiography, categorize receptor subtypes, and probe mechanisms of receptor signaling, via measurements of agonist binding and its regulation by ions, nucleotides, and other allosteric modulators. This unit reviews the theory of receptor binding and explains how to analyze experimental data. Since binding data are usually best analyzed using nonlinear regression, this unit also explains the principles of curve fitting with nonlinear regression. PMID- 18428566 TI - Fear-potentiated startle in mice. AB - Pavlovian fear conditioning is frequently used to assess the behavioral, physiological, genetic and molecular correlates of learning and memory. In the typical Pavlovian conditioned fear procedure a neutral stimulus, such as a tone, is paired with a mildly aversive stimulus such as a foot shock. The tone conditioned stimulus (CS) comes to elicit a variety of behaviors that are indicative of learned fear. One of the more prominent of these behaviors is a potentiated acoustic startle response. While fear-potentiated startle in mice is qualitatively similar to that in rats, the stimulus parameters and procedures for producing optimum fear-potentiated startle in mice differ considerably from those used in rats. Procedures outlined in this unit include initial assessment of startle, fear conditioning and fear-potentiated startle testing. Special attention is paid to the parameters that affect the magnitude of fear-potentiated startle and procedures designed to systematically examine these parameters are included. PMID- 18428567 TI - Ultrasonic vocalizations in rat and mouse pups. AB - The ultrasonic vocalization (USV), or isolation calling response, of infant rats and mice has been studied as a measure of the intensity of an aversive affective state and as an early communicative behavior between pup and mother. The four protocols described in this unit are for the basic isolation testing procedure, and for elicitation of the contact quieting response to littermates and/or dam, the potentiation of isolation calling response by a prior brief maternal interaction and the predator-induced suppression of USV by the scent of an unfamiliar male. These procedures for the elicitation of USV, and for its regulation by different kinds of social interaction, provides the basis for experimental research on the early development of emotion and communication in an animal model system. PMID- 18428568 TI - Parental behaviors in rats and mice. AB - Parental care in mammals is often critical for survival and development of the offspring. The multiple behaviors that comprise parental care in rats and mice are controlled by a complex array of factors. This unit describes the components of parental care in rats and mice and the frequently used methods to assess the performance of these behaviors, a subject's preference for pup-related cues, as well as a subject's motivation to act parentally. PMID- 18428569 TI - Overview of rodent models for obesity research. AB - Animal obesity models differ widely in type and extent of obesity. They are either based on environmental factors (e.g., high-fat diet-induced obesity), spontaneous mutants (i.e., ob/ob mice), genetically engineered animals (e.g., mice with melanocortin receptor subtype-4 gene disruption (knock-out), or mechanical intervention (e.g., chemical lesion of the ventromedial hypothalamus). This unit reviews available rodent models to study obesity and attempts to highlight the greatest utility for each model. PMID- 18428570 TI - Intravenous self-administration of ethanol in mice. AB - A more complete understanding of alcohols reinforcing actions is obtained when multiple behavioral procedures are used, some of which bypass taste factors. This unit describes a method for assessing the reinforcing effects of alcohol in mice using the most widely accepted procedure for assessing drug reward: intravenous self-administration. PMID- 18428571 TI - Preclinical models to evaluate potential pharmacotherapeutic agents in treating alcoholism and studying the neuropharmacological bases of ethanol-seeking behaviors in rats. AB - The unit outlines four basic protocols designed to systematically evaluate the capacity of potential pharmacotherapeutic agents to effectively treat alcohol addiction and dependence in rats. Also included are procedures designed to study the neural mechanisms regulating alcohol-seeking behaviors. PMID- 18428572 TI - Models of amyotrophic lateral sclerosis. AB - Amyotrophic lateral sclerosis (ALS) is a progressive neurological disorder caused by degeneration of the motor neurons in cortex, brainstem and spinal cord. Two experimental models of ALS are described in this unit: organotypic cultures of spinal cord, and transgenic mice expressing a human mutant superoxide dismutase 1 (SOD1) gene. Appropriate animal and cell culture models of ALS can be used to help unravel the sequence of events in motor neuronal degeneration and test potential therapies. PMID- 18428573 TI - A murine model of retroviral encephalopathy. AB - This unit delineates the steps for production of a murine model of retroviral encephalopathy. The LP-BM5 infected mouse develops a chronic inflammation of the brain secondary to profound immune deficiency. The model is robust, develops rapidly and does not require the use of human pathogens. In addition, the behavioral and neurochemical characteristics of this model is reviewed. PMID- 18428574 TI - Animal models of retroviral encephalopathies: feline model. AB - Human immunodeficiency virus infection in children and adults results in a progressive neurodegenerative disease consistent with a predominant subcortical mediated dementia. Techniques for developing a feline model of the early stages of lentiviral-associated neurodegeneration are presented. The behavioral, neurophysiologic, immunologic, virologic, and neuropathologic aspects of this model are also described. PMID- 18428575 TI - Internet resources for genomic, bioinformatics, and medical genetics information. AB - Many diseases are influenced by multiple genetic and environmental factors. Identifying the genes contributing to the probability of developing such diseases poses an extraordinary challenge because each gene may have a small influence. Further, in the presence of other influential genes and environmental factors, the impact of a single gene may be amplified. Many approaches are being taken to address the challenge presented by complex trait genetics, and data are being amassed at an alarming rate. Access to these data is crucial for coordination of efforts and avoidance of unnecessary duplication of research. This appendix describes some of the resources available on the World Wide Web that provide genetic and genomic data, tools for analyzing genome data, information on funding opportunities, and information about ethical, legal and social issues associated with the genetic analysis of disease traits. PMID- 18428576 TI - Basic neuroanatomical methods. AB - This unit covers some of the basic procedures that are common to a wide range of neuroanatomical protocols. Procedures are provided for the preparation of unfixed, fresh brain tissue as well as for perfusion fixation of animals resulting in fixed neural tissue. A variety of methods for sectioning brains are described, including frozen sectioning in a cryostat, frozen sectioning with a microtome, and sectioning with a vibratome. The choice of sectioning method depends on how the brain has been prepared and what histochemical method is to be used. Three post-sectioning procedures are provided: defatting of slide-mounted sections, thionin staining of the sections, and coating of slides with photographic emulsion for autoradiography. Finally, a procedure is described for subbing slides with gelatin, which is necessary in some protocols in order for the sections to adhere to the slides. PMID- 18428577 TI - Quantitative in situ hybridization for the study of gene expression at the regional and cellular levels. AB - Quantitative in situ hybridization allows measurement of mRNA level modifications in a variety of experimental conditions. This analysis may be performed both at the regional anatomical and cellular levels by densitometry, neuronal counting and silver grain measurements. PMID- 18428578 TI - Counting cells in sectioned material: a suite of techniques, tools, and tips. AB - This unit presents protocols to obtain accurate estimates of cell density and cell number in sectioned material by using a light microscope. The "optical disector" or "3-D counting method" is described, followed by Abercrombie's less commonly used two-section comparison (TSC) method. These basic protocols are accompanied by four support protocols: one for celloidin embedding, which renders superb morphology, one for point counting, which is important for volume measurements and is almost always used in conjunction with the disector or 3-D counting, one for handling the potential problem of z-axis distortion and the consequences that this error can have on density estimates and sampling tactics when using the disector, and finally, one that provides a guide for calibrating and verifying estimates obtained by counting methods. PMID- 18428579 TI - Production of a heterozygous mutant cell line by homologous recombination (single knockout). AB - Gene targeting by homologous recombination allows the introduction of specific mutations into any cloned gene. This unit provides a protocol in which the gene of interest is inactivated by interrupting its coding sequence with a positive selectable marker. A negative selectable marker is included in the construct outside the region of target gene homology in order to enrich for clones in which the target gene has undergone homologous recombination. The altered target gene is then expressed in embryonic stem cells. A support protocol describes a method for transient expression of Cre recombinase to remove sequences between lox sites, which can also be used as a selection method. PMID- 18428580 TI - RNA analysis by nuclease protection. AB - Nuclease protection assays (S1 nuclease protection and RNase protection) are extremely sensitive procedures for detection and quantitation of mRNA species in complex mixtures of total cellular RNA. These assays are well suited for mapping positions of external and internal junctions in RNA, such as transcription initiation and termination sites and intron/exon boundaries, and to discriminate between closely related targets by using probes designed to span the regions where the related genes differ the most. Also, because the size of the probes used in nuclease protection assays is a variable chosen by the investigator, probes may be designed to protect fragments of different sizes. This feature permits the simultaneous analysis of several different mRNAs in the same total RNA sample. In this unit, a method is included for RNase protection of target mRNA sequences, including hybridization of the probe to the target sequence, details of the actual protection assay, and detection of reaction products. An alternative method is provided for performing the RNase protection assay on a microvolume scale, which is useful when there are many samples to be analyzed. Support protocols describe synthesis and gel purification of labeled RNA probes; preparation of RNase-free yeast RNA, which acts as an aid in the quantitative precipitation of newly synthesized probe; and quantitation of target mRNA. A method describing S1 nuclease protection of target mRNA using either RNA or DNA probes is also included. Additional support protocols provide instructions for the preparation of radiolabeled DNA probes by primer-extension of double-stranded plasmid or PCR product using Klenow fragment of E. coli DNA polymerase I or Taq or Tth polymerase in a thermal cycler. Another radiolabeling method details 5' end labeling of oligodeoxynucleotides and oligoribonucleotides using T4 polynucleotide kinase. Additionally, a method is described for mapping transcription start sites using the S1 nuclease protection assay. PMID- 18428581 TI - Chronic recording of extracellular neuronal activity in behaving animals. AB - Two methods for recording extracellular neuronal activity in unanesthetized, unrestrained rats are described in this unit. Both use chronically-implanted bundles of fine microwires to record electrophysiological activity. One method provides recordings of single and/or multiple unit activity from individual wires in a bundle (monotrode). Discrimination of individual neuronal potentials is based on action potential amplitude, or on a combination of action potential amplitude and shape. The second method uses a 2- to 4-microwire array (stereotrode-tetrode) to yield multiple unit recordings. Discrimination of individual neuronal potentials is based on action potential shapes and the relative amplitude of action potentials recorded simultaneously on the different wires in the array. These methods can provide stable, long-term recording of neuronal activity during a variety of behavioral paradigms. PMID- 18428582 TI - Imaging nervous system activity with voltage-sensitive dyes. AB - Optical recording with a voltage-sensitive dye is advantageous where membrane potential must be recorded in many sites at once. This unit describes methods for making voltage-sensitive dye measurements on different preparations to study (1) how a neuron integrates its synaptic input into its action potential output by measuring membrane potential everywhere synaptic input occurs and where spikes are initiated; (2) how a nervous system generates a behavior in Aplysia abdominal ganglion; and (3) responses to sensory stimuli and generation of motor output in the vertebrate brain by simultaneous measurement of population signals from many areas. The approach is three-pronged: (1) find the dye with the largest signal-to noise ratio; (2) reduce extraneous sources of noise; and (3) maximize the number of photons measured to reduce the relative shot noise. A discussion of optical recording methods including the choice of dyes, light sources, optics, cameras, and minimizing noise is also provided. PMID- 18428583 TI - Mouse social recognition and preference. AB - Social recognition in mice is represented by a simple pattern of behavior that can be accurately and reliably quantified by trained observers. The paradigm presented in this unit takes advantage of an ethologically relevant phenomenon marked by a vigorous and species-typical sequence of investigatory behaviors that occurs when conspecifics meet. Recognition is noted by decreased investigation of a previously encountered animal. PMID- 18428584 TI - Assessment of murine startle reactivity, prepulse inhibition, and habituation. AB - The study of the startle response and its plasticity provides a powerful research paradigm for examining the neural control of behavior. The neural substrates underlying phenomena such as information-processing functions, including sensorimotor gating, affective modulation, and habituation, can be studied using startle plasticity as an operational measure. The mouse is rapidly becoming the most powerful model system for the study of human CNS function and is highly amenable to both genetic and pharmacological study. This unit describes procedures for the assessment of the startle response to acoustic stimuli in mice; methods for assessing prepulse inhibition (PPI) of startle, with treatments including drug administration and genetic manipulation; and methods for testing habituation. PMID- 18428585 TI - Social learning of food preferences in rodents: rapid appetitive learning. AB - A procedure is described for quantitating the transfer of information about inducing increased intake of distinctively-flavored foods or fluids among common laboratory rodents. The method provides a simple, efficient, non-invasive way to produce robust, long-lasting changes in appetitive behaviors of laboratory rodents that can be used in studies of the neuroanatomical, neurochemical or genetic substrates of learning and memory. PMID- 18428586 TI - Assessment of sustained and divided attention in rats. AB - Behavioral tasks must be evaluated in terms of the cognitive functions they require in order to be performed. All of the tasks described in this chapter can be used with each of four experimental manipulations: stimulation of a single brain region by drugs or small electrical current, impairment of normal function by production of a lesion or administration of appropriate pharmacological agents, recording of brain activity during the performance of a specific behavioral task, or behavioral phenotyping of transgenic and knockout mice for genes expressed in specific brain regions. This unit describes protocols for the radial arm maze task and the water maze task, both of which require intact spatial memory abilities. PMID- 18428587 TI - Models of neuropathic pain in the rat. AB - There are now three models of neuropathic pain in the rat that are in widespread use: the chronic constriction injury, the partial sciatic ligation model, and the spinal nerve ligation model. The procedures to create these models and the behavioral assays used to quantify the resulting abnormal pain sensations are described in this unit. PMID- 18428588 TI - Place preference test in rodents. AB - Conditioned place preference is a behavioral model currently used to measure the rewarding properties induced by the administration of a drug. In this paradigm, the rewarding properties of a compound are associated with the particular characteristics of a given environment. Due to the recent availability of multiple lines of genetically modified mice, this unit includes procedures for testing place preference in both rats and mice. PMID- 18428589 TI - Inducing photochemical cortical lesions in rat brain. AB - In the rat photochemical cortical lesion model described in this unit, an intravascular photochemical reaction induces endothelial damage resulting in platelet aggregation, thrombosis, thrombotic response (secretion of factors by the platelets) and permanent cerebral vascular occlusion. Because thrombosis is produced in pial vessels, the resulting cortical infarct is generally smaller and more reproducible than in the models involving occlusion of the middle cerebral artery. The surgical procedures involved are limited, making this model generally easier to perform and less invasive than most other models of permanent focal ischemia that involve mechanical occlusion of major cerebral arteries. PMID- 18428590 TI - Measurement of panic-like responses following intravenous infusion of sodium lactate in panic-prone rats. AB - This unit describes a putative animal model for panic disorder. The basic premise is that pharmacological disruption of critical brain regions implicated in the circuitry of anxiety will lead to a condition similar to that of the human disorder. A clinically relevant test, the sodium lactate challenge, is utilized to assess parallels between the human condition and this rat model. PMID- 18428591 TI - Handling and restraint. AB - For the safety of the handler and the animal, proper methods for handling and restraining laboratory animals should be followed. Improper handling can result in increased stress and injury to the animal. In addition, the handler risks injury from bite wounds or scratches inflicted when the animal becomes fearful or anxious. By using sure, direct movements with a determined attitude, the animal can be easily handled and restrained. Animals can be restrained either manually or in a plastic restrainer. The protocols in this unit describe handling and manual restraint of mice, rats, hamsters, and rabbits. Alternate protocols describe restraint using the plastic restrainer. PMID- 18428592 TI - Animal identification. AB - Proper identification of animals is crucial in research, whether the animals are being used as experimental subjects, for breeding purposes, or as a source for tissues, cells, or fluids. This unit provides specific protocols for the ear punch (notch) and ear tag methods for mouse, rat and hamster; tattoos for mouse, rat and rabbit; and the correct use of a subcutaneous transponder system for all of the above species. PMID- 18428593 TI - Databases and Web sites for neurogenetics. AB - The goal of neurogenetics is an understanding of the genetic basis of brain structure and function. Neurogenetics deals with multiple areas of investigation, including the genetic basis of neural induction, patterning, cell fate specification, neuron maturation, axonal and dendritic organization, synapse function, and learning and behavior. This appendix provides links to databases and other Web sites used by neurobiologists for recovery of information about genes involved in neural development and neuron function. Special care has been taken to curate sites involving model organisms, since neurogenetics relies heavily on comparative genomics to establish gene function. PMID- 18428594 TI - Intracellular and juxtacellular staining with biocytin. AB - Many physiological studies require microscopic examination of the recorded neuron for identification. This unit describes how intracellular and extracellular recording can be combined with single-neuron staining to enable sequential physiological and morphological studies. PMID- 18428595 TI - Quantified assessment of terminal density and innervation. AB - Stereological methods allow for the determination of cell numbers, terminal densities, and, subsequently, the estimation of terminal arbor size within a given brain nucleus. This unit provides an explanation for determining the terminal arbor size of dopaminergic neurons of the nigrostriatal pathway in rodents. In contrast to previously used single-axon reconstructions, these stereological methods allow for quick and easy determination of terminal arbor size. PMID- 18428596 TI - Immunohistochemical localization of proteins in the nervous system. AB - The immunohistological methods described in this unit can be used to determine the precise localization of neurochemicals, receptors, and proteins throughout the nervous system. Determining the localization of a protein within defined brain nuclei and neuronal cell populations can provide important clues regarding its potential function. Immunoperoxidase reactions and light microscopy are commonly used to visualize the distribution of a single primary antibody directed to an antigen of interest. Double-labeling immunofluorescence and confocal microscopy techniques detect the localization of one protein relative to another protein and allow analysis of colocalization at a cellular and subcellular level. The colocalization of two proteins can also be quantified, allowing analysis of the extent of overlap between two labeled markers and measurements of changes in the localization of one protein relative to another following drug treatment or in animals that have been genetically modified. The theoretical limit of resolution of confocal microscopy is 0.1 to 0.2 microm. PMID- 18428597 TI - Hybridization histochemistry of neural transcripts. AB - Expression of genes is manifested by the production of RNA transcripts within cells. Hybridization histochemistry (or in situ hybridization) permits localization of these transcripts with cellular resolution or better. Furthermore, the relative amounts of transcripts detected within different tissues or the same tissues in different states (e.g., physiological or developmental) may be quantified. This unit describes hybridization histochemical techniques using either oligonucleotide probes or RNA probes (riboprobes). Also presented is the use of probes labeled with digoxigenin for colorimetric detection of RNA transcripts and a technique to detect the Y chromosome using either mouse or human riboprobes. Finally, a procedure is presented for the autoradiographic detection of radiolabeled probes. Methods are provided for labeling oligodeoxynucleotide and RNA probes and performing northern analyses using these probes. PMID- 18428598 TI - Multidisciplinary approaches for characterizing synaptic vesicle proteins. AB - Investigation of synaptic vesicle membrane proteins using multidisciplinary approaches, particularly to characterize synaptic vesicle proteins in synapses, can greatly advance our knowledge of the molecular mechanisms involved in synaptic vesicle exocytosis and neurotransmission. Three approaches are presented in this unit to identify and characterize synaptic vesicle proteins. The first is a subcellular fractionation method used to isolate synaptic vesicles from rat brain synaptosomal preparations, which can then be used in a variety of biochemical studies on synaptic vesicle proteins. The second is a detailed procedure for pre-embedding immunogold staining and electron microscopic observation, techniques that permit the morphological identification of synaptic vesicle proteins in individual vesicles at the intact synapse. Finally, a protocol for immunocytochemical staining of cultured hippocampal neurons for light microscopic examination is provided, which allows one to stain multisynaptic vesicle proteins and determine their localization in relation to other proteins or subcellular structures in synapses. PMID- 18428599 TI - Long-term culture of hippocampal neurons. AB - In culture, hippocampal cells can develop to express neuronal antigens and acquire mature neuronal morphologies, including axons, complex dendritic trees, and synapses that are electrophysiologically active. This system is suitable for studying neuronal differentiation and other events, such as synaptogenesis. It is also a valuable model for investigating synaptic plasticity and exploring the mechanisms of neuronal degeneration. This unit provides a protocol for culturing neurons prepared from embryonic (E-18) rat or mouse hippocampus, but could also be used to grow neurons from embryonic cortex, olfactory bulb, striatum, or spinal cord. A second method is included for preparing neuronal cultures from embryos with different genotypes, such as those from transgenic mice. Also described is the preparation of polyornithine- and fibronectin-coated coverslips, which are highly adhesive and promote neurite outgrowth, for use in the culture protocols. PMID- 18428600 TI - Protein expression in the Drosophila Schneider 2 cell system. AB - The Schneider-2 (S2) Drosophila cell line is well suited for the stable overexpression of recombinant proteins using plasmid-based protein expression vectors. Following drug selection, a polyclonal S2 cell line can be induced to express on the order of 2 to 100 pmol/mg membrane protein for G-coupled protein receptors, 4000 to 100,000 sites/cell for other membrane receptors and 3 to 35 mg/liter for soluble and secreted proteins. PMID- 18428601 TI - Immunoblotting and immunodetection. AB - Immunoblotting (western blotting) is used to identify specific antigens recognized by polyclonal or monoclonal antibodies. This unit provides numerous protocols for all steps starting with solubilization of the protein samples, usually with SDS and reducing agents. Following solubilization, the material is separated by SDS-PAGE and the antigens are then electrophoretically transferred to a membrane, a process that can be monitored by reversible staining or Ponceau S staining. The transferred proteins are bound to the surface of the membrane, providing access to immunodetection reagents. Any remaining binding sites are blocked by immersing the membrane in a blocking solution. After probing with the primary antibody, the membrane is washed and the antibody-antigen complexes are identified with horseradish peroxidase (HRPO) or alkaline phosphatase enzymes coupled to the secondary anti-IgG antibody (e.g., goat anti-rabbit IgG) and appropriate chromogenic or luminescent substrates. Finally, membranes may be stripped and reprobed. PMID- 18428602 TI - Recording in the cerebellar slice. AB - This unit describes techniques for the preparation of mouse cerebellar slices and electrophysiological recording from neurons in the slice. The cerebellum provides a model of motor learning that can be correlated with alterations in synaptic function. In addition, the architecture and well-defined synaptic pathways in the cerebellar slice make this preparation a useful model for exploring general principles of synaptic transmission. PMID- 18428603 TI - Patch-clamp recording from neuronal dendrites. AB - Pyramidal neurons of the central nervous system have extensively arborized apical dendrites that contribute importantly to the signaling properties of the neuron. Recent advances in electrophysiological techniques have allowed recording from neuronal dendrites. These techniques depend on using infrared optics to visualize dendritic processes in the unstained brain slice preparation, on pipet positioning with high resolution micromanipulators, and on stringent techniques for brain slice preparation that preserved healthy dendritic processes, even in tissue from mature animals. The procedures underlying these techniques are described in this unit. PMID- 18428604 TI - Fabrication of patch pipets. AB - Patch clamping refers to a wide range of electrophysiological measurements, all of which have in common the use of patch pipets and the formation of gigaohm seals. The purpose of this unit is to describe the fabrication of patch pipets. The aspects of the pipet geometry that are important to different applications and the different procedures that have been found to most reliably and simply achieve these results are described. Parameters for glass selection are detailed in the beginning of the unit. Pulling patch and whole-cell pipets, elastomer coating, fire polishing, pipet filling, and pipet testing in an experimental setup are highlighted. Additional support protocols describe alternative ways to optimize pipet geometry and cleaning the glass before pulling. Considerations for noise and dynamic performance are emphasized as these two requirements for single channel and whole-cell current measurements dictate how the pipets must be fabricated. PMID- 18428605 TI - Assessment of developmental milestones in rodents. AB - Developmental tests are used to characterize early markers of behavior for investigation of the neurobiology of these behaviors, and to assess the impact of early prenatal or postnatal insult. These perturbations may include pharmacological, environmental, and genetic manipulations. At birth the rat is capable of some specific activities, but its movements are uncoordinated and seemingly random, its tactile sensitivity is not fully developed, and its ear canals and eyes remain closed until several days after birth. Postnatal development consists mainly of the continuation of processes begun earlier. This unit presents protocols for the most commonly used animal tests of developmental reflexology, including negative geotaxis, cliff avoidance, placing responses, tactile and acoustic startle responses, surface and air righting reflexes, crossed extensor reflex, rooting reflex, grasp reflex, bar holding, and horizontal and vertical screen tests. PMID- 18428606 TI - Animal tests of anxiety. AB - Animal tests of anxiety are used to screen novel compounds for anxiolytic or anxiogenic activity, to investigate the neurobiology of anxiety, and to assess the impact of other occurrences such as exposure to predator odors or early rearing experiences. This unit presents protocols for the most commonly used animal tests of anxiety. The Geller-Seifter conflict test, the social interaction test, light/dark exploration, the elevated plus-maze, defensive burying, and the thirsty rat conflict. The protocols are described in terms of drug screening tests, but can be modified easily for other purposes. PMID- 18428607 TI - Assessment of spatial memory using the radial arm maze and Morris water maze. AB - Behavioral tasks must be evaluated in terms of the cognitive functions they require in order to be performed. All of the tasks described in this chapter can be used with each of four experimental manipulations: stimulation of a single brain region by drugs or small electrical current, impairment of normal function by production of a lesion or administration of appropriate pharmacological agents, recording of brain activity during the performance of a specific behavioral task, or behavioral phenotyping of transgenic and knockout mice for genes expressed in specific brain regions. This unit describes protocols for the radial arm maze task and the water maze task, both of which require intact spatial memory abilities. PMID- 18428608 TI - Cued and contextual fear conditioning in mice. AB - Contextual and cued fear conditioning is a robust form of learning in which an association is made between stimuli and their aversive consequences. Fear conditioning has been used in laboratory rodents in part because it is a highly conserved form of behavior that is exhibited in both laboratory situations and in normal environments. Training requires only a single trial and this makes it adaptable to genetic, pharmacological, and biochemical studies. Clinically, it is has relevance to human behavior in that fear conditioning can be produced in humans, and damage to the amygdala prevents fear conditioning. PMID- 18428609 TI - Assessment of learning and memory using the autoshaping of operant responding in mice. AB - This unit describes the use of an automated procedure for developing an operant response ("autoshaping") in the mouse. The method has applications in the study of the acquisition of behavior (learning) as well as for the assessment of memory or retention of that task. PMID- 18428610 TI - Social transmission of food preference in mice. AB - The social transmission of food preference (STFP) is an ethologically relevant test of olfactory memory that can be used in mice. In this test, "observer" mice interact with a "demonstrator" mouse that has recently eaten a novel food. When observer mice are presented with a choice between the food eaten by the demonstrator and some other novel food, observer mice prefer the food eaten by the demonstrator. This phenomenon depends on the observer mice detecting olfactory cues on the breath of the demonstrator mouse during their interaction. The subsequent food preference serves as a measure of memory for those olfactory cues. This unit describes a method for performing STFP in mutant mice. The method explains how to address potential confounding factors and avoid possible pitfalls. PMID- 18428611 TI - Direct, continuous behavioral analysis of drug action on feeding. AB - Analysis of rodent feeding behavior during the study of anorectic drugs reveals that contextual variables such as type of food, drug, environment, and physiological state influence food intake. Temporal changes in behavior can signal the onset of a feeding modulation process, as is the case with anorectic drugs acting on distinct neurochemical mechanisms that change temporal behavioral profiles. Comparing the effects of anorectic agents in controlled animal assays can identify potential therapeutic drugs that may influence human eating behavior. This unit specifies the materials and procedures required for continuously monitoring the Behavioral Satiety Sequence, as well as a device for continuous, automatic measurement of the mass of food consumption by the animal during behavioral monitoring called the RUEMA (Rat Universal Eating Monitor Apparatus). PMID- 18428612 TI - Conditioned flavor aversions: assessment of drug-induced suppression of food intake. AB - Administration of a drug following ingestion of a novel food or solution often suppresses subsequent intake of the new food or solution. This suppression is associative, in that consumption is not suppressed when there is no temporal relationship between consumption and drug administration. The robust nature of aversion learning has made this procedure a sensitive and widely used behavioral index of drug side effects. The procedures described in this unit are suitable for work with rodents, and may require modifications, e.g., in presentation of the ingesta and drug for other species. Familiar and novel foods may be used instead of solutions, with similar results. PMID- 18428613 TI - Macronutrient selection in experimental animals. AB - To address the question of how animals choose diets from an array of nutritionally different foods, researchers have designed experiments in which animals are provided with separate sources of the three macronutrients, protein, fat and carbohydrate. These experiments are useful for investigating the neuroanatomical and neurochemical mechanisms involved in food choice, as well as for assessing the effects of changes in physiological status (e.g., pregnancy; lactation and increased energy expenditure) or disease states (e.g., diabetes; and obesity) on nutrient intakes. PMID- 18428614 TI - Animal models of painful diabetic neuropathy: the STZ rat model. AB - Painful peripheral neuropathy is a common secondary complication of diabetes. The streptozotocin (STZ)-induced diabetic rat is the most commonly employed animal model used to study mechanisms of painful diabetic neuropathy and to evaluate potential therapies. A low dose STZ protocol is described for inducing experimental diabetes in the rat. Several behavioral assays are described, which are routinely used to assess different aspects of neuropathic pain in this animal model of diabetes mellitus, including mechanical allodynia and heat hyperalgesia. PMID- 18428615 TI - Traumatic brain injury in the rat using the fluid-percussion model. AB - Traumatic brain injury is a leading cause of death and disability, particularly among young adults. During closed head trauma, the injury process is initiated by the impact of the brain against the inner table of the calvarium. Subsequently, there is prompt initiation of a complex biochemical, cellular, and physiological injury cascade that may take days to complete. From a functional standpoint, this culminates in neurologic dysfunction and, if severe, death. This unit describes an impact-induced brain trauma model in rats which replicates nonpenetrating head injury. It does not model either penetrating or ischemic brain injuries. PMID- 18428616 TI - Parenteral injections. AB - This unit describes the techniques for the following routes of injection for mice, rats, hamsters and rabbits: intramuscular, intradermal, subcutaneous, intravenous, intraperitoneal, footpad, and intrathymic. Guidelines are also given regarding injection volumes and temperatures, and the use of proper restraints. PMID- 18428617 TI - Blood collection. AB - Blood is most frequently sampled for evaluation of serum antibodies or analysis of surface markers on peripheral blood cells. The protocols offered in this unit describe collection of blood from the orbital sinus or plexus of the mouse, rat, or hamster. With appropriate techniques, small amounts of blood can be obtained with little ill effect on the animal. Collection from the auricular vein or artery of the rabbit is also relatively unstressful to the animal. Bleeding procedures that should be performed on the anesthetized animal include collection from the mouse axillary plexus, cardiac puncture of the mouse, rat, hamster, or rabbit, and collection from the hamster abdominal aorta or vena cava. PMID- 18428618 TI - Euthanasia. AB - Euthanasia of laboratory animals must be performed by trained personnel using appropriate techniques, equipment, and reagents in order to effect a death that is humane and satisfies research requirements. Acceptable methods of euthanasia are painless or minimize distress, and are quick and easy to perform, safe for those performing the procedure, and efficient and economic. They are aesthetically acceptable and are done in the absence of other animals. In addition, these methods do not result in gross histological or histochemical changes that would adversely affect research results. This unit offers protocols for euthanasia employing carbon dioxide asphyxiation (see Basic Protocol 1), pentobarbital overdose (see Basic Protocol 2), exsanguination, and cervical dislocation for the mouse, rat, hamster, and rabbit. PMID- 18428619 TI - 3-D reconstruction of neurons from multichannel confocal laser scanning image series. AB - A confocal laser scanning microscope (CLSM) collects information from a thin, focal plane and ignores out-of-focus information. The operator configures separate channels (laser, filters, detector settings) for each fluorochrome used in a particular experiment. Then, 3-D reconstructions are made from Z-series of confocal images: one series per channel. Channel signal separation is extremely important and measures to avoid bleaching are vital. Post-acquisition deconvolution of the image series is then performed to increase resolution. In the 3-D reconstruction program described in this unit, reconstructions can be inspected in real time from any viewing angle. By altering viewing angles and by switching channels off and on, the spatial relationship of 3-D-reconstructed structures with respect to structures seen in other channels can be studied. Since each brand of CLSM, computer program, and 3-D reconstruction package has its own proprietary set of procedures, a general approach is provided wherever possible. PMID- 18428620 TI - Two-photon imaging in live rodents. AB - Two-photon imaging is an innovative optical technique that has quickly become state of the art for imaging fluorescent signals in a variety of organisms. With many advantages over conventional confocal microscopy, such as greater image resolution, deeper access (approximately 400 microm), and much less photo damage, two-photon microscopy has already proven to be an extremely useful tool for imaging live cells or tissue. Due to its tremendous versatility, recent efforts have adapted this technique to allow visualization of fluorescent cells directly in living animals. This unit describes a basic procedure for performing two photon imaging in vivo as applied to the dorsal surface of the brain in live anesthetized mice or rats. The protocol outlines a surgical preparation to enable the capture of stable, high-resolution (<1 microm) images of fluorescently labeled neurons in intact brain with very little detrimental effect to either cells or tissue. PMID- 18428621 TI - Fluorescence-based sorting of neural stem cells and progenitors. AB - Neural stem cells (NSCs) are defined as undifferentiated cells originating from the neuroectoderm that have the capacity both to perpetually self-renew without differentiating and to generate multiple types of lineage-restricted progenitors (LRPs). LRPs can themselves undergo limited self-renewal and ultimately differentiate into highly specialized cells that make up the nervous system. However, this physiologically delimited definition of NSCs and LRPs has become increasingly blurred due to lack of protocols for effectively separating these types of cells from primary tissues. This unit discusses recent attempts using fluorescence-activated cell sorting (FACS) strategies to prospectively isolate NSCs from different types of LRPs as they appear in vivo, and details a protocol that optimally attains this goal. Thus, the strategy presented here provides a framework for more precise studies of NSC and LRP cell biology in the future, which can be applied to all vertebrates, including humans. PMID- 18428622 TI - An overview on the generation of BAC transgenic mice for neuroscience research. AB - This unit provides a comprehensive overview on the generation of transgenic mice using bacterial artificial chromosomes (BACs), and the application of BAC transgenic mice in neuroscience research. In the first section, advantages of the BAC transgenic approach compared to the conventional transgenic approach are summarized. In the second section, important considerations in designing BAC transgenic constructs are outlined. Four commonly used BAC transgenic construct designs are also outlined. Concepts of modifying BACs by homologous recombination in E. coli to introduce a variety of mutations into BACs, and important steps to characterize a modified BAC prior to the generation of transgenic mice are also presented. In the final section, some of the important applications of BAC transgenic mice in neuroscience research, including studying gene expression, gene function, mapping neuronal circuitry, and modeling human diseases, are described. PMID- 18428623 TI - Modification of bacterial artificial chromosomes (BACs) and preparation of intact BAC DNA for generation of transgenic mice. AB - BAC transgenesis is a powerful tool for the study of gene expression and gene function in the mouse in vivo. In this unit, detailed protocols are provided for modification (i.e., marker gene insertion, deletion, or point mutation) of BACs by homologous recombination in E. coli. This method utilizes a shuttle vector that allows transient expression of the E. coli RecA gene to support homologous recombination in the BAC host bacteria. In addition, two protocols are provided for purification of BAC DNA for microinjection to generate transgenic mice. Since BAC DNA is prone to degradation, which may introduce positional effects in transgenic mice, two methods are given for purification of intact BAC DNA for subsequent microinjection. PMID- 18428624 TI - Scintillation proximity assay. AB - Scintillation proximity assay technologies provide a rapid non-separation method to measure common biological interactions using radioactively tagged molecules. This unit identifies potential uses of the technology for the measurement of receptor-ligand binding, cAMP accumulation, GTP binding to heterotrimeric G proteins, protease activity and cellular uptake. PMID- 18428625 TI - Defensive responses to predator threat in the rat and mouse. AB - Defensive responses include an array of specific behaviors, including flight, freezing, risk assessment, and defensive threat/attack, that are elicited by unconditioned threat stimuli such as predators or predator odors. Some individual defensive behaviors are selectively responsive to drugs effective against generalized anxiety disorder or panic, providing a rationale for their use in investigation of compounds that may be useful in treating these disorders. In addition, defensive behaviors toward predators and some predator odors show rapid conditioning to contextual stimuli, whereas other predator odors do not, although they too elicit defensiveness. This pattern suggests that the ability of a predator odor to predict danger may be a determinant of the degree to which that odor supports aversive conditioning. Predators and predator odors are also increasingly used in studies of brain systems potentially related to emotionality. These factors indicate the need for selective, reliable, and convenient tests of defensiveness to predators and predator odors using rat and mouse subjects. PMID- 18428626 TI - Olfactometry with mice. AB - A computer-controlled, multiple-channel odor generator, operant test chamber, and discrimination training procedure for mice are described. The odor generator allows controlled presentation of any one or combinations of eight odors to an odor-sampling port that contains a liquid reinforcement delivery tube that also serves to detect responses. A modified discrete trial operant conditioning procedure provides measures of both response accuracy and response rate during anticipation of stimulus delivery and in the presence of two different odor stimuli. Results from numerous experiments demonstrate that, with these methods, mice reliably and rapidly acquire odor detection and a variety of odor discrimination tasks, and that response rate in the presence of the odor reflects the incentive motivation of stimuli that are or are not associated with reward. PMID- 18428627 TI - Assaying aspects of attention and impulse control in mice using the 5-choice serial reaction time task. AB - First developed in the 1980s by Trevor Robbins in Cambridge to investigate attentional function in rats, the 5-choice serial reaction time task involves continuous scanning by the subject across a spatial array of visual stimuli. On detecting a brief stimulus at one of five locations, the subject must make a nose poke in order to collect a reward. The task has also been conducted successfully in several strains of mice, including transgenic models. This unit presents the procedures required to run the task in mice, outlining the expected results at each stage of training and presenting examples of the most common manipulations used to dissociate behavior further. PMID- 18428628 TI - Chemoconvulsant model of chronic spontaneous seizures. AB - Animal models of injury-induced epilepsy may provide insight into the mechanisms of acquired epilepsy. Previous animal models of temporal lobe epilepsy (TLE) were produced by acute treatments that often have high mortality rates and/or are associated with a low proportion of animals developing spontaneous, chronic motor seizures. In this unit, a protocol is provided for inducing chronic epilepsy in rats using multiple, low-dose, intraperitoneal injections of an excitotoxic agent, kainic acid. This protocol reliably induces TLE in nearly all treated rats (97% had at least two observed spontaneous motor seizures) with a relatively low mortality rate (<15%). This modified chemoconvulsant treatment protocol (i.e., multiple low doses) is efficient and relatively simple, and the properties of the chronic epileptic state appear similar to those of severe human TLE. PMID- 18428629 TI - Chronic intravenous drug self-administration in rats and mice. AB - Chronic intravenous drug self-administration in rodents is a useful procedure for predicting the abuse liability of novel drugs in humans, for evaluating candidate treatments for drug abuse and dependence, and for studying the biological basis of addiction. This unit focuses on recent technical innovations for conducting long-term studies of i.v. drug self-administration behavior in healthy, freely moving rats and mice. Included are protocols for construction, implantation, and maintenance of chronic indwelling jugular catheters, commentary on critical parameters, a troubleshooting guide, and anticipated results. PMID- 18428630 TI - Intravenous self-administration techniques in monkeys. AB - Drug self-administration is a procedure in which a subject performs a response, called an operant, that results in the delivery of a drug injection. This procedure is viewed as a relevant model for the study of human drug-taking behavior. Drug self-administration in primates has several characteristics that resemble drug-taking behavior in humans, and drugs that are commonly abused by humans also typically maintain self-administration behavior in monkeys. Drug self administration procedures allow for the study of a variety of drug properties. For instance, they are used to investigate the abuse potential of new compounds and to study the effects of candidate medications for the treatment of drug addiction. These procedures also can be used to study the process of drug reinforcement. This unit describes intravenous drug self-administration in large primates, such as rhesus macaques, and smaller primates, such as squirrel monkeys. PMID- 18428631 TI - Overview on rodent models of Alzheimer's disease. AB - In Alzheimer's disease (AD), characteristic lesions develop in brain regions that subserve cognitive functions, ultimately leading to dementia. There are now several lesioned or transgenic small-animal models of the disease that model select aspects of cognitive deficits and/or recapitulate many, but not all, of the characteristic pathologic lesions observed in AD. This overview describes the most common approaches used to model AD in rodents, highlights their utility, and discusses some of their deficiencies. PMID- 18428632 TI - Anterograde axonal tract tracing. AB - The mammalian brain contains a myriad of interconnected regions. An examination of the complex circuitry of these areas requires sensitive neuroanatomical tract tracing techniques. The anterograde tracers, Phaseolus vulgaris leucoagglutinin (PHA-L) and biotinylated dextran amines (BDA) are powerful tools that can be used to label fiber tracts that project from one particular brain region. When injected iontophoretically, PHA-L and BDA are readily taken up by neurons and transported anterogradely along their axonal tracts. Combined with immunocytochemistry for neurotransmitters, neuropeptides, and receptors, tract tracing methods may be used to elucidate the phenotype of synapses that form the microcircuitry of specific neural systems. PMID- 18428633 TI - Whole-mount immunohistochemistry of the brain. AB - The gross anatomical distribution of an antigen is typically mapped using a combination of serial sectioning, immunocytochemistry, and three-dimensional reconstruction. This is a tedious and time-consuming procedure, which introduces an array of potential alignment and differential shrinkage errors and requires considerable experience and specialized equipment. In particular, it is unsuited for routine screening applications. To circumvent these problems, this unit presents a routine whole-mount immunocytochemistry protocol that can be used to map many antigenic distributions in the developing and adult brain. The technique can also be easily adapted to detect anterograde and retrograde transport tracers. PMID- 18428634 TI - Rodent sensory neuron culture and analysis. AB - Sensory neurons have proven very useful for analysis of neuronal differentiation in vivo and in vitro. Their utility for in vitro work is based on the fact that sensory neurons are relatively easy to isolate in large numbers and are amenable to manipulations in culture. Lumbar ganglia are usually used because their location in the caudal nervous system means they are the least differentiated at any developmental stage, allowing the analysis of relatively undifferentiated cells. Rodent sensory ganglia from embryonic to adult stages can be dissected effectively and maintained in serum-free medium or in coculture with other cells or factors. This unit describes generation of embryonic rat lumbar dorsal root ganglia (DRG) cultures, which form an important model system for investigating the cellular and molecular mechanisms that regulate neuronal differentiation. Adult DRG can also be successfully cultured, with a few modifications of the general protocol. PMID- 18428635 TI - Quantitative analysis of in vivo cell proliferation. AB - Injection and immunohistochemical detection of 5-bromo-2'-deoxyuridine (BrdU) has become the standard method for studying the birth and survival of neurons, glia, and other cell types in the nervous system. BrdU, a thymidine analog, becomes stably incorporated into DNA during the S-phase of mitosis. Because DNA containing BrdU can be specifically recognized by antibodies, this method allows dividing cells to be marked at any given time and then identified at time points from a few minutes to several years later. BrdU immunohistochemistry is suitable for cell counting to examine the regulation of cell proliferation and cell fate. It can be combined with labeling by other antibodies, allowing confocal analysis of cell phenotype or expression of other proteins. The potential for nonspecific labeling and toxicity are discussed. Although BrdU immunohistochemistry has almost completely replaced tritiated thymidine autoradiography for labeling dividing cells, this method and situations in which it is still useful are also described. PMID- 18428636 TI - Production of recombinant adeno-associated viral vectors and use for in vitro and in vivo administration. AB - Adeno-associated virus is a nonpathogenic human virus that has been developed into a gene-delivery vector due to its high efficiency of infection for many different cell types and its ability to persist and lead to long-term gene expression. This unit describes efficient methods to generate high-titer, research-grade, adenovirus-free recombinant single-stranded and self complementary adeno-associated virus in various serotypes, along with methods to quantify the viral vectors. Two detailed methods are provided for viral vector injection into rat brain and for histological detection of transgene expression, including lacZ and GFP. PMID- 18428637 TI - Production and titration of lentiviral vectors. AB - Lentiviral vectors have emerged over the last decade as powerful, reliable and safe tools for stable gene transfer in a wide variety of mammalian cells. Unlike other vectors derived from oncoretroviruses, they allow for stable gene delivery into most nondividing primary cells, including neurons. This is why LVs are becoming the most useful and promising tools in the field of neuroscience, not only for research, but also for future gene and cell therapy approaches. Lentivectors (LVs) derived from HIV-1 have gradually evolved to display many desirable features aimed at increasing both their safety and their versatility. These latest designs are reviewed in this unit. This unit also describes protocols for production and titration of LVs that can be implemented in a research laboratory setting, with an emphasis on standardization to improve transposability of results between laboratories. PMID- 18428638 TI - Imaging protein-protein interactions by Fluorescence Resonance Energy Transfer (FRET) microscopy. AB - Detection of specific protein-protein interactions has long been restricted to bulk biochemical methods such as immunoprecipitation and immunoblotting. Even more sensitive methods using general immunofluorescence are limited, and it is difficult to infer protein-protein interactions from the results of these tests. Fluorescence Resonance Energy Transfer (FRET) is a photophysical process that can be exploited to obtain highly sensitive information about such interactions. It can sense the presence of acceptor fluorophores in the vicinity of a donor fluorophore within a separation distance that is the size of a single protein molecule. This unit details FRET microscopy based on release of quenched donor fluorescence after acceptor photobleaching, microinjection of reagents into the nucleus or cytosol, and labeling of antibodies for these procedures. PMID- 18428639 TI - Monitoring protein-protein interactions in living cells by bioluminescence resonance energy transfer (BRET). AB - Bioluminescence resonance energy transfer (BRET) allows monitoring of protein protein interactions in real time in living cells. One candidate interacting protein is fused to a luminescent energy donor, such as Renilla luciferase, and the other to a fluorescent energy acceptor, such the green fluorescent protein (GFP), and the two are then coexpressed in the same cells. If the two proteins interact, their close proximity allows nonradiative energy transfer (BRET) between the luciferase and the GFP. BRET does not occur if the two proteins are separated by more than 100 A, making the technique ideal for monitoring protein protein interactions in biological systems. This unit describes the use of BRET to study constitutive and agonist-promoted interactions among signaling molecules, as illustrated by the homodimerization of the CXCR4 receptor and the recruitment of beta-arrestin2 to agonist-activated G-protein-coupled receptors. This noninvasive and homogeneous assay provides a robust and sensitive proteomic platform with applications for basic science research and drug discovery. PMID- 18428640 TI - Immunoprecipitation. AB - Immunoprecipitation is a technique in which an antigen is isolated by binding to a specific antibody attached to a sedimentable matrix. It is also used to analyze protein fractions separated by other biochemical techniques such as gel filtration or density gradient sedimentation. The source of antigen for immunoprecipitation can be unlabeled cells or tissues, metabolically or intrinsically labeled cells, or in vitro-translated proteins. This unit describes a wide range of immunoprecipitation techniques, using either suspension or adherent cells lysed by various means (e.g., with and without detergent, using glass beads, etc.). Flow charts and figures give the user a clear-cut explanation of the options for employing the technology. PMID- 18428641 TI - Detection of protein-protein interactions by coprecipitation. AB - Coprecipitation of proteins from whole-cell extracts is a valuable approach to testing for physical interactions between proteins of interest. When a precipitating antibody is used, this method is referred to as co immunoprecipitation. Coprecipitation can be used to study interactions between known proteins and as a means of identifying components of a complex. This unit describes basic approaches to immunoprecipitating tagged proteins from whole-cell extracts. The extract is prepared under nondenaturing conditions, the protein of interest is precipitated, and the precipitate is tested for a second specifically associated protein. The approach can be used for native or epitope-tagged proteins for which antibodies are available, or for recombinant proteins that bind with high affinity to a molecule that can be coupled to a solid-phase matrix. An associated protein is detected by separating the precipitated proteins by SDS-PAGE and immunoblotting with a second antibody that recognizes the putative associated protein. PMID- 18428642 TI - Caged molecules: principles and practical considerations. AB - A caged molecule is an inert but photosensitive molecule that is transformed by photolysis into a biologically active molecule at high speed (typically 1 msec). The process is referred to as photorelease. The spatial resolution of photorelease is limited by the properties of light; submicrometer resolution is potentially achievable. Therefore, focal photorelease of caged molecules enables one to control biological processes with high spatio-temporal precision. The principles underlying caged molecules as well as practical considerations for their use are discussed in this unit. PMID- 18428643 TI - Photolysis of caged neurotransmitters: theory and procedures for light delivery. AB - Photolysis of "caged" compounds is a technique for releasing biologically active compounds in which the timing, rate, and spatial profile of release are controlled by light. Issues relating to the delivery of light for single-photon photolysis are presented. Specific discussions include the theories relating to how light interacts with biological tissue to produce scattering and phototoxicity, as well as the issues involved in choosing the appropriate light source. Several approaches and optical designs are presented for delivering the output of a laser to a microscopic specimen. The criteria for choosing an approach are presented. The commercial sources for the parts needed to build a photolysis system are also provided. This unit will be particularly useful for investigators interested in single-photon photolysis of caged neurotransmitters in brain slices. PMID- 18428644 TI - Overview of methods for assessing the mouse auditory system. AB - The three general approaches for assessing the mouse auditory system presented in UNITS 8.21B-8.21D are complementary, but differ in the time required for testing as well as the goals of the tests: the auditory brainstem response (ABR; UNIT 8.21B), distortion product otoacoustic emissions (DPOAE; UNIT 8.21C), and lick suppression behavioral test (UNIT 8.21D). Various issues that need to be addressed in the testing of mouse hearing are discussed in this overview, including accurate nomenclature of the strain and/or mutations, environmental versus genetic influences, selection of frequencies to be tested, and appropriate animal care and use. PMID- 18428645 TI - Measurement of the auditory brainstem response (ABR) to study auditory sensitivity in mice. AB - The ABR is an electroencephalographic response measured with scalp electrodes. It provides a quick, easy, and reliable method for physiological assessment of auditory sensitivity in mice. A series of brief tone pips or clicks is presented to an anesthetized mouse at a high rate of speed; each click evokes waves of neural activity in the brainstem that are computer-averaged so they are differentiated from non-auditory background voltages. The intensity of the clicks is reduced in steps until an ABR can no longer be discerned, thereby defining the ABR threshold, which is closely related to the hearing threshold. Key procedural issues are: (1) accurate calibration of the acoustics (what sounds arrive at the mouse's ear), (2) anesthetization of the mouse, (3) setting up the recording electrodes, (4) the protocol for presenting acoustic stimuli and obtaining thresholds, and (5) interpretation of ABR data. PMID- 18428646 TI - Assessment of cochlear function in mice: distortion-product otoacoustic emissions. AB - Distortion-product otoacoustic emissions (DPOAEs) can be measured in the ear canal following the presentation of two tones. These emissions are generated by the outer hair cells (OHCs) of the inner ear and they are reduced or absent when the OHCs are damaged by, for example, exposure to excessive noise or ototoxic drugs. Consequently, DPOAEs provide a powerful and noninvasive means to assess the robustness of OHC function. A detailed method is described for measuring DPOAEs to assess cochlear function in mice. Recommendations are given for the required equipment and instructions are presented for setting up a DPOAE system. Also, a protocol is outlined for measuring DPOAEs in mice and troubleshooting tips are provided. Examples of data analysis procedures following noise exposure in mice are included, as well. These methods are not only applicable to mice, but can be performed using essentially all small laboratory animals. PMID- 18428647 TI - Behavioral assessment of hearing in mice--conditioned suppression. AB - The method of conditioned suppression described in this unit involves training a thirsty mouse to make steady contact with a waterspout in order to receive a slow, but steady trickle of water and then pairing a sound with mild electric shock delivered through the spout. The mouse quickly learns to avoid the shock by breaking contact with the spout whenever it detects the sound. This suppression of drinking is then used to indicate that the animal detected the sound. Because it is a cognitively simple procedure, conditioned suppression can be used to determine the ability of brain-damaged and genetically altered as well as normal animals to detect and discriminate sounds. PMID- 18428648 TI - Application of experimental stressors in laboratory rodents. AB - This unit presents eight separate stressor protocols for laboratory rodents. Stress induction is a critical element in the study of neural and neuroendocrine mechanisms involved in establishing and maintaining a state of stress. The first four procedures, immobilization, footshock, swimming, and noise, involve acute exposure to noxious stimuli. The next three procedures, social isolation, resident/intruder aggression, and maternal deprivation, induce social disruption by withdrawal from a group housing condition, attack within the unfamiliar territory of a dominant male, or segregation of a preweanling pup from its mother, respectively. The final procedure, sleep deprivation, involves passive denial of the opportunity to sleep. Support protocols are provided to address the need for environmental acclimation and calming procedures prior to any stress related studies (including, for rats, handling of the animals as a calming measure) and to detail a simple method of quantifying the response to a given stressor by direct measurement of levels of the stress hormones adrenocorticotropic hormone (ACTH) and corticosterone. PMID- 18428649 TI - Binge eating in rats produced by combining dieting with stress. AB - This unit describes a rodent model of binge eating based on cyclic restriction, refeeding, footshock, and intermittent access to palatable food. These conditions mimic dieting, stress, and "junk" food indulgence, respectively, all common etiological and maintenance factors in human binge eating. Four groups of rats are used: one subjected to cyclic food restriction, another to acute footshock stress, another to both of these (R + S), and a control. Neither cyclic restriction nor stress alone produces binge eating, but the R + S rats, despite satiety, double their intake of palatable food in a discrete period of time (i.e., binge) when stressed. This protocol recapitulates critical properties of human binge eating, namely preference for palatable food, dieting- and stress induced vulnerability to binging, and eating for reward versus metabolic need. This protocol permits study of the psychobiological underpinnings of binge eating and possibly also of addiction, impulsivity, and depression, which are co-morbid with binge eating. PMID- 18428650 TI - Binge eating in rats with limited access to vegetable shortening. AB - In this protocol, binge-type eating is induced in non-food-deprived rats by providing limited access to an optional source of dietary fat: vegetable shortening. The protocol is simple and inexpensive, and the binge behavior is robust, reliable, and maintainable across extended periods of time. Two peptides that normally affect fat intake in rats have no effect on fat intake under limited-access conditions. However, recent results with a GABA(B) receptor agonist and with progressive-ratio responding suggest that the behavior induced by the limited-access binge protocol may share similarities with substance abuse. This protocol is designed to model the kind of excessive behavior that characterizes bingeing-related eating disorders and certain addictions. PMID- 18428651 TI - Sugar bingeing in rats. AB - Bingeing behavior is characteristic of many eating disorders. This unit describes an animal model of sugar bingeing. This model has been used successfully to elicit behavioral and neurochemical signs of sugar dependence in rats, e.g., indices of bingeing, withdrawal, increased intake after abstinence (deprivation effect), cross-sensitization with amphetamine, and increases in dopamine release in the nucleus accumbens due to repeated bingeing. PMID- 18428652 TI - Characterization of energy expenditure in rodents by indirect calorimetry. AB - The need for treatment of obesity and obesity-related diseases, such as type 2 diabetes, has been intensified by the epidemic rise of obesity. Recent advances make possible continuous monitoring of metabolically relevant functions in animals to identify novel thermogenic and anorectic compounds. This unit describes non-invasive in vivo calorimetric assessment of energy expenditure using measurements of oxygen consumption and carbon dioxide production, complemented by telemetric monitoring of body core temperature and locomotor activity in mice and rats. Reference compounds are used to illustrate the determination of substance-specific parameters, such as the dose that produces the half-maximal effect (ED(50)), the maximal effect, as well as the time of onset and duration of compound action. Indirect calorimetry performed at different temperatures provides information on several other well-defined parameters, including resting metabolic rate, basal metabolic rate, lower critical temperature, temperature sensitivity, defended body temperature, and respiratory quotient. PMID- 18428653 TI - Techniques for mammalian cell tissue culture. AB - This unit opens with detailed discussions on the latest principles of sterile technique and preparation of culture media. Step-by-step protocols describe trypsinizing and subculturing monolayer cultures, passaging suspension cultures, freezing and thawing cells, counting cells using a hemacytometer, and preparing cells for transport. PMID- 18428654 TI - Histochemical methods for the detection of apoptosis in the nervous system. AB - Neuroscientists often need to detect neuron death at the light microscope level in tissue sections derived from animal models of neurological disease. In many instances there is a need to detect apoptosis, the most common morphology of programmed cell death. This unit provides two protocols for the detection of apoptosis by immunostaining for either activated forms of caspases or their cleavage products. When used in conjunction with nuclear dyes, these protocols permit visualization not only of caspase activation, but also the nuclear chromatin clumps characteristic of apoptosis. The first protocol utilizes peroxidase-mediated chromogen deposition to visualize antibodies by brightfield microscopy. The second protocol utilizes fluorophores to visualize antibodies by epifluorescence. Double immunofluorescence labeling can be performed to identify the phenotype of cells in which caspases are activated. Not all cell death is apoptotic. Therefore, a third protocol is presented for suppressed silver staining, a useful method to screen for all morphologic forms of cell death. PMID- 18428655 TI - Gene delivery using helper virus-free HSV-1 amplicon vectors. AB - Herpes simplex virus type 1 (HSV-1)-based amplicon vectors contain only a very small percentage of the 152-kbp viral genome. Consequently, replication and packaging of amplicons depend on helper functions that are provided either by replication-defective mutants of HSV-1 or by replication-competent, but packaging defective, HSV-1 genomes. Sets of cosmids that overlap and represent the entire HSV-1 genome can form, via homologous recombination, circular replication competent viral genomes, which give rise to infectious virus progeny. However, if the DNA cleavage/packaging signals are deleted, reconstituted virus genomes are not packageable, but still provide all the helper functions required for the packaging of cotransfected amplicon DNA. The resulting stocks of packaged amplicon vectors are essentially free of contaminating helper virus. This unit describes the cotransfection of amplicon and cosmid or bacterial artificial chromosome (BAC) DNA into 2-2 cells by cationic liposome-mediated transfection using and the harvesting of packaged vector particles. Support protocols provide methods for preparing cosmid DNA and determining the titers of amplicon stocks. PMID- 18428656 TI - Alphaviruses: Semliki Forest virus and Sindbis virus vectors for gene transfer into neurons. AB - Alphaviral vectors based on Semliki Forest virus and Sindbis virus infect many host cell types, causing rapid and high-level transgene expression. Compared to other viruses used to infect CNS cell and tissue preparations, Semliki Forest virus and Sindbis virus exhibit an outstanding preference for neurons rather than glial cells. High-titer vector generation typically requires biosafety level 1 or 2 containment and occurs in less than 2 days. Wild-type vectors are cytotoxic, permitting short-term transgene expression. However, mutant vectors with decreased cytotoxicity, to prolong host cell survival, have been developed. They also increase transgene expression and cellular coinfection, permitting heteromeric protein expression in individual cells. Other mutants with temperature-dependent control of transgene expression and altered host cell preference to target interneurons and astrocytes rather than principal neurons are available. Because of these advantages, alphaviral vectors are increasingly used in neurobiological and other studies, including structural biology, vaccine development, and cancer treatment. PMID- 18428657 TI - Overview of gene targeting by homologous recombination. AB - The analysis of mutant organisms and cell lines is important in determining the function of specific proteins. Recent technological advances in gene targeting by homologous recombination in mammalian systems enable the production of mutants in any desired gene, and can be used to produce mutant mouse strains and mutant cell lines. The yeast Flp/FRT recombinase system and bacteriophage recombinases such as Cre and its recognition sequence, loxP, allow spatial and temporal control of knockouts. This unit discusses crucial issues for homologous recombination experiments, including requirements for the source of DNA, criteria for the targeting constructs, methods of enrichment for homologous recombinants, (positive and negative selection, and the use of endogenous promoters), and the types of mutations that can be created. PMID- 18428658 TI - Lentiviral strategies for RNAi knockdown of neuronal genes. AB - RNA interference (RNAi) refers to the process by which 21- to 23-nucleotide short interfering RNAs (siRNAs) mediate post-transcriptional degradation of homologous mRNA transcripts. This process is carried out by an endogenous pathway that centers on the use of endogenously encoded small RNAs, and can be hijacked to knock down the expression of any target protein by introducing a specific siRNA into a cell. Stable knockdown can be obtained by constitutive expression of the siRNA from the host chromosome. Retroviruses, such as lentivirus, provide a convenient vector by which to integrate RNAi expression constructs. Lentiviruses can infect nondividing cells, thereby allowing knockdown in cells such as mature neurons. This unit provides methods to design and clone siRNAs into a lentiviral vector. Additional protocols describe production and titering of the lentivirus, as well as safety testing. Finally, methods are provided for infecting neurons in culture and in vivo with RNAi lentivirus. PMID- 18428659 TI - Split-ubiquitin system for identifying protein-protein interactions in membrane and full-length proteins. AB - Protein-protein interactions play a fundamental role in the regulation of almost all cellular processes. Thus, the identification of interacting proteins can help to elucidate their function. The mating-based split-ubiquitin system (mbSUS) uses yeast as a test organism to identify potential interactions between full-length membrane proteins or between a full-length membrane protein and a soluble protein. The mbSUS can also be used to provide further evidence for protein protein interactions detected with other methods and to map the interaction domains of selected proteins. The mbSUS is optimized for systematic screening approaches employing a mating-based approach, as typically used to determine protein interactions on a genomic scale. Construction of bait and prey fusions is simplified by adapting two different cloning procedures: (i) in vivo cloning in yeast, and (ii) Gateway cloning in E. coli. Protocols for small-scale interaction tests, as well as systematic approaches using sorted bait and prey arrays, are described. PMID- 18428660 TI - Production of antisera using fusion proteins. AB - This unit details the use of bacterially produced fusion proteins for the production of antisera, allowing for the large-scale generation of affinity purified antibodies to specific, targeted epitopes. The use of pET vectors containing a polyhistidine (His) or glutathione-S-transferase (GST) tag to construct bacterial expression plasmids are provided as prototypical examples of fusion protein methodology. The basic protocols provided in this unit describe: (1) transformation of E. coli for high-yield production of soluble fusion protein, (2) purification of soluble fusion proteins for use in immunization using chelated nickel or glutathione affinity chromatography (for His- and GST tagged fusion proteins, respectively), (3) immunization of rabbits with purified fusion protein and collection of antisera, and (4) characterization of antisera for antibody specificity using immunoblotting techniques. Support protocols describe the purification of His-tagged insoluble fusion proteins for animal immunization and the construction and use of affinity columns for purifying antibodies using soluble fusion proteins. PMID- 18428661 TI - Whole-cell recording in vivo. AB - In vivo whole-cell patch-clamp recording provides a means for measuring membrane currents and potentials from individual cells in the intact animal. Patch-clamp methods have largely been developed in vitro. This body of work has contributed enormously to the understanding of many important phenomena in excitable cells- including synaptic plasticity in the mammalian central nervous system, and the behavior of individual protein channels. In recent years, an increasing number of groups have applied whole-cell recording techniques in the intact animal. Such in vivo studies offer the tantalizing possibility of uncovering the underlying principles and mechanisms of neural interactions within the natural context of fully intact biological networks. This unit focuses on strategies for overcoming the specific technical challenges posed by in vivo whole-cell recording. A straightforward procedure is described for obtaining whole-cell records from the cortex of the anesthetized rat; this procedure has also been applied successfully to awake animals and other rodent species with minor modifications. PMID- 18428662 TI - Electrophysiological recordings from neonatal neocortical brain slices. AB - Many brain processes and the expression of many neuronal proteins are developmentally regulated; therefore, studying cellular and molecular processes in tissue from very young animals is vital to understanding the development of the brain. Working on tissue from very young animals (first postnatal week) presents a number of specific problems. This unit describes techniques to overcome these problems to produce in vitro slices from neonatal rodents and to reliably achieve high-quality patch-clamp recordings from neurons in neonatal brain slices. PMID- 18428663 TI - Assays for cyclic nucleotide-specific phosphodiesterases (PDEs) in the central nervous system (PDE1, PDE2, PDE4, and PDE10). AB - Since the identification of phosphodiesterase activity in brain tissue more than 40 years ago, 11 distinct gene families have been identified, differing with respect to localization, regulation, affinity for cAMP and cGMP, and distinct functions within cells. PDEs 1, 2, 4, and 10 are currently of special interest to CNS pharmacology because of their high expression in specific areas of the brain and the behavioral effects of inhibitors of these enzymes in rodents. Efficient high-throughput PDE enzyme assays are essential for PDE-targeted drug discovery, and this unit details two types of assays. The first method is relatively inexpensive and is based on separating radiolabeled cNMPs from degradation products on alumina columns. The second method is fluorescence-based; it is fast and better accommodates high-throughput screening, but is more expensive. Although these methods have successfully been used for PDEs 1, 2, 4 and 10, they could be readily adapted to other PDEs. PMID- 18428664 TI - Locomotor behavior. AB - The study of locomotor activation in rodents does not involve extensive learning or conditioning, and so this dependent measure is often used as the initial screen for pharmacological effects predictive of therapeutic efficacy of a drug class in humans. Methods for evaluating locomotor behavior in rodents fall into two general categories: automated monitoring of behavior and direct observational techniques. This unit presents protocols for measurement of locomotor activity using photocell-based automated monitoring systems, and direct observation using interval and ordinal scales. Other methods used to quantify locomotor activity include video-based systems, rotometers, and running wheels; these are discussed in the commentary. The protocols provided are designed for rats, although they can easily be adapted for use with mice and other rodents. PMID- 18428665 TI - Delay discounting and impulsive choice in the rat. AB - Delay discounting refers to the degree to which immediate outcomes exhibit more influence over behavior than outcomes which are delayed. Impulsive choice, in the context of delay discounting, is generally considered as an increased preference for immediate over delayed outcomes, even where the delayed outcomes are more advantageous. In the past decade, there has been increasing use of delay discounting paradigms to elucidate the physiological, pharmacological, and behavioral aspects of the putative neural circuitry underlying impulsive choice. This unit describes the assessment of impulsive choice in the rat using a delay discounting procedure involving an operant response choice between a small reinforcer delivered immediately and a larger reinforcer delivered after a delay, which is progressively increased within a session. Variations of some of the main task parameters are also discussed, as well as their significance and interpretation. PMID- 18428666 TI - Models of nociception: hot-plate, tail-flick, and formalin tests in rodents. AB - Experimental models of pain include tests of response thesholds to high intensity stimuli (acute pain tests) and changes in spontaneous or evoked behavioral responses in animals with peripheral injury or inflammation (persistent pain models). Acute thermal pain is modeled by the hot-plate and tail-flick test, while persistent pain can be modeled by the formalin test. This unit presents protocols for all three of these tests, including preparation of animals (rats or mice), administration of a compound being tested for its analgesic properties and data collection. PMID- 18428667 TI - Overview of animal models of schizophrenia. AB - Animal models of schizophrenia may increase the understanding of the neurological abnormalities associated with the disorder and aid in the development of rational pharmacological treatments. Rather than attempting to model the entire syndrome of schizophrenia, a more biologically oriented approach to animal models has been to focus on specific symptoms of schizophrenia that are more objectively measured in the clinical population and more directly translatable to animals (e.g., observables or endophenotypes). This overview focuses on behavioral measures that have been investigated in rodent models of schizophrenia with varying degrees of predictive, etiological, and construct validity. Because of the severity of cognitive deficits in schizophrenia and their resistance to current treatments, there is a need to develop animal models specific to the cognitive symptoms of schizophrenia. In light of this need, this overview discusses rodent models of cognition with relevance to the core cognitive deficits observed in schizophrenia. PMID- 18428668 TI - Ratings of L-DOPA-induced dyskinesia in the unilateral 6-OHDA lesion model of Parkinson's disease in rats and mice. AB - This unit provides detailed protocols for establishing rodent models of L-DOPA induced dyskinesia. The 6-hydroxydopamine (6-OHDA) lesion procedure is described in more detail for mice than for rats since the lesioning procedure in rats has been described extensively in previous work and is less difficult to perform. Unlike primate models, rodent models of L-DOPA-induced dyskinesia are relatively simple and fast to set up, thus being affordable to most laboratories. These models allow for studying the dyskinetic complications of L-DOPA treatment on large groups of animals under strictly controlled experimental conditions. Along with information and structured protocols for the practical execution of the test, this unit provides a detailed description of the rating scale and the phenomenology of rodent abnormal involuntary movements, and suggestions for beginners. PMID- 18428669 TI - Mouse and rat anesthesia and analgesia. AB - Many animal models used in neuroscience research must be surgically created and/or anesthetized for imaging studies. The purpose of this unit is to review the advantages and disadvantages of various anesthetic and analgesic agents in rodents; to discuss state-of-the-art methods for monitoring anesthesia; and to provide tips for troubleshooting problems with anesthesia. PMID- 18428670 TI - Biochemical fractionation of brain tissue for studies of receptor distribution and trafficking. AB - An important tool for studying the regulation of synapses is a rapid and reliable means of separating synaptic and intracellular proteins. This unit presents a technique for analysis of brain tissue which relies on differential centrifugation to separate proteins present at synaptic sites from those found in intracellular cytoplasmic and vesicular pools. The method is efficient in that only small amounts of tissue, such as might be obtained from a small region of a rodent brain, are required. It is reproducible and, in conjunction with immunoblot or immunoprecipitation techniques, can produce reliable quantitative data. The protocol will be of interest to those conducting a variety of different studies related to the localization and trafficking of brain receptors and signaling molecules. PMID- 18428671 TI - Neural stem cell transplantation in mouse brain. AB - Neural stem cells (NSCs) are the most primordial, least committed cells of the nervous system, and transplantation of these multipotent cells holds the promise of regenerative therapy for many central nervous system (CNS) diseases. This unit describes methods for NSC transplantation into neonatal mouse pups, embryonic mouse brain, and adult mouse brain. A description of options for detection of labeled donor cells in engrafted mouse brain is provided along with an example protocol for detecting lacZ-expressing cells in situ. Also included is a protocol for preparing NSCs for transplantation. PMID- 18428672 TI - Overview of mouse assays of ethanol intoxication. AB - There are many behavioral assays to assess sensitivity to ethanol intoxication in mice. Most are simple to implement, and are sensitive to a particular dose range of ethanol. Most reflect genetic influences, and each test appears to reflect the contribution of a relatively distinct collection of genes. This genetic heterogeneity implies that no single test can claim to capture the construct "ethanol intoxication" completely. Depending on the test, and when measurements are made, acute functional tolerance to even a single dose of ethanol must be considered as a contributing factor. Whether or not a test is conducted in naive mice or as part of a test battery can influence sensitivity, and do so in a strain-dependent manner. This unit reviews existing tests and recommends several. PMID- 18428673 TI - Retrograde axonal tracing with fluorescent markers. AB - The growth of fluorescence imaging technology and the development of sensitive fluorescent retrograde tracers has provided many new approaches for analyzing neuronal circuits. Fluorescent markers provide unparalleled opportunity for combining axonal tract tracing with techniques such as immunohistochemistry or physiological recording. This unit describes the use of six different fluorescent tracers: Fast Blue, fluorescein dextran, FluoroGold, FluoroRuby, red beads, and green beads. Guidance is provided on how to choose a tracer for a particular experiment, and three methods are described for injecting the tracers, including pressure injection through a microsyringe or a micropipet, and iontophoretic injection through a micropipet. Criteria for selecting the most appropriate method are discussed. The protocols provide the information necessary to take advantage of the numerous fluorescent tracers that are available and to apply them to a wide variety of scientific questions. PMID- 18428674 TI - Ballistic labeling with fluorescent dyes and indicators. AB - Neuronal cell labeling is fundamental to investigations of the nervous system. Labeling of cells in live or fixed tissue with dyes or ion indicators using ballistic approaches has recently been developed for the study of neuronal architecture and function. In this approach, dye-coated particles are propelled into cells by a pulse of pressurized helium. This unit provides step-by-step protocols for coating tungsten particles with fluorescent or indicator dyes and for delivering these particles into cells and tissue. The major advantage of the ballistic method of dye delivery is that large populations of neurons can be rapidly labeled within a piece of live or fixed tissue. Advantages and limitations of the approach are discussed and technical advice is provided. PMID- 18428675 TI - Use of FM1-43 and other derivatives to investigate neuronal function. AB - The fluorescent dye FM1-43 and its derivatives can be used to monitor the physiology of synaptic vesicle turnover in central nerve terminals. They do so by their ability to reversibly partition into membranes, a process that results in a huge increase in fluorescence in comparison to their quantum yield in solution. This unit provides protocols for quantifying total synaptic vesicle turnover, the kinetics and extent of synaptic vesicle exocytosis, and the kinetics and mode of synaptic vesicle endocytosis. Descriptions of other ways these protocols have been used to derive information about the life cycle of the synaptic vesicle are also provided. PMID- 18428676 TI - Dissection and culture of embryonic spinal commissural neurons. AB - Studies of spinal commissural neurons have provided substantial insight into the mechanisms that regulate axon guidance. Explants of embryonic spinal cords and isolated spinal commissural neurons have been important experimental tools for the identification and characterization of several guidance cues, including netrins, semaphorins, slits, sonic hedgehog, BMPs, and wnts. In this unit, protocols are provided for establishing these explant assays to assess the outgrowth and turning capacity of commissural axons. In addition, methods are included for preparing cultures highly enriched with embryonic commissural neurons, which have been used to probe the biochemical signaling mechanisms regulating axon guidance. PMID- 18428677 TI - Schedule-induced polydipsia: a rat model of obsessive-compulsive disorder. AB - Obsessive-compulsive disorder (OCD) is difficult to model in animals due to the involvement of both mental (obsessions) and physical (compulsions) symptoms. Due to limitations of using animals to evaluate obsessions, OCD models are limited to evaluation of the compulsive and repetitive behaviors of animals. Of these, models of adjunctive behaviors offer the most value in regard to predicting efficacy of anti-OCD drugs in the clinic. Adjunctive behaviors are those that are maintained indirectly by the variables that control another behavior, rather than directly by their own typical controlling variables. Schedule-induced polydipsia (SIP) is an adjunctive model in which rats exhibit exaggerated drinking behavior (polydipsia) when presented with food pellets under a fixed-time schedule. The polydipsic response is an excessive manifestation of a normal behavior (drinking), providing face validity to the model. Furthermore, clinically effective drugs for the treatment of OCD decrease SIP. This protocol describes a rat SIP model of OCD and provides preclinical data for drugs that decrease polydipsia and are clinically effective in the treatment of OCD. PMID- 18428678 TI - Using the iHOP information resource to mine the biomedical literature on genes, proteins, and chemical compounds. AB - The iHOP (Information Hyperlinked over Proteins) resource employs sophisticated text-mining methods to assist researchers in their quest for information on specific genes and proteins, their physical interactions and regulatory relationships, their relevance in pathologies, and their interactions with chemical compounds. iHOP parses thousands of PubMed documents every day and collects information specific to thousands of different biological molecules. Rather than providing long lists of entire abstracts upon keyword searches, iHOP selectively retrieves information that is specific to genes and proteins, and summarizes their interactions and functions. In iHOP, with genes and proteins acting as hyperlinks between sentences and abstracts, a large part of the PubMed knowledgebase becomes a giant navigable information network. PMID- 18428679 TI - Using the MetaCyc pathway database and the BioCyc database collection. AB - The MetaCyc database (http://metacyc.org) is a collection of more than a thousand metabolic pathways from a wide variety of organisms, collected from the literature by manual curation. BioCyc contains more than 370 Pathway/Genome Databases, each of which describes the genome and predicted metabolic pathways of a single organism, as computationally predicted from the annotated genomes, using MetaCyc data as a reference. The protocols in this unit introduce the user to the extensive data content available in MetaCyc and BioCyc, and to mechanisms for querying, visualizing, and analyzing these data using the Pathway Tools software. PMID- 18428680 TI - Using the tools and resources of the RCSB protein data bank. AB - The Protein Data Bank (PDB; http://www.pdb.org) is the world-wide repository for three-dimensional structural data determined using various experimental methods. The options and procedures for searching and downloading structural data from the Research Collaboratory for Structural Bioinformatics (RCSB) PDB are described here, along with tools for assessing the quality of structures. Several types of information are associated with each structure deposition, including atomic coordinates of the structure, experimental data used to solve it, sequences of all macromolecules in the structures, details about the structure solution method, images showing different views of the structure, derived geometric data, and a variety of links to other resources. These data and resources may be used for understanding the function and stability of the molecule and for designing biochemical, genetic, or other experiments. They can also be used for molecular modeling and drug design. PMID- 18428681 TI - Using BiblioSpec for creating and searching tandem MS peptide libraries. AB - BiblioSpec is a software package for creating and searching libraries of tandem MS peptide spectra. Library searching provides a quick method for making peptide spectrum matches by comparing a query spectrum to a collection of reference spectra of known peptide sequence. Pre-assembled libraries for several model organisms can be used as the basis of a search or custom libraries can easily be assembled. The protocols in this unit describe installing BiblioSpec, searching libraries, and creating custom libraries. PMID- 18428682 TI - Using N-SCAN or TWINSCAN to predict gene structures in genomic DNA sequences. AB - N-SCAN is a gene-prediction system that combines the methods of ab initio predictors like GENSCAN with information derived from genome comparison. It is the latest in the TWINSCAN series of programs. This unit describes the use of N SCAN to identify gene structures in eukaryotic genomic sequences. Protocols for using N-SCAN through its Web interface and from the command line in a Linux environment are provided. Detailed discussion about the appropriate parameter settings, input-sequence processing, and choice of genome for comparison are included. PMID- 18428683 TI - Using the Proteomics Identifications Database (PRIDE). AB - The Proteomics Identifications Database (PRIDE) is a public data repository designed to store, disseminate, and analyze mass spectrometry based proteomics datasets. The PRIDE database can accommodate any level of detailed metadata about the submitted results, which can be queried, explored, viewed, or downloaded via the PRIDE Web interface. The PRIDE database also provides a simple, yet powerful, access control mechanism that fully supports confidential peer-reviewing of data related to a manuscript, ensuring that these results remain invisible to the general public while allowing referees and journal editors anonymized access to the data. This unit describes in detail the functionality that PRIDE provides with regards to searching, viewing, and comparing the available data, as well as different options for submitting data to PRIDE. PMID- 18428684 TI - Using GFS to identify encoding genomic loci from protein mass spectral data. AB - Genome-based peptide fingerprint scanning (GFS) directly maps several types of protein mass spectral (MS) data to the loci in the genome that may have encoded for the protein. This process can be used either for protein identification or for proteogenomic mapping, which is gene-finding and annotation based on proteomic data. Inputs to the program are one or more mass spectrometry files from peptide mass fingerprinting and/or tandem MS (MS/MS) along with one or more sequences to search them against, and the output is the coordinates of any matches found. This unit describes the use of GFS and subsequent results analysis. PMID- 18428685 TI - Using TESS to predict transcription factor binding sites in DNA sequence. AB - This unit describes how to use the Transcription Element Search System (TESS). This Web site predicts transcription factor binding sites (TFBS) in DNA sequence using two different kinds of models of sites, strings and positional weight matrices. The binding of transcription factors to DNA is a major part of the control of gene expression. Transcription factors exhibit sequence-specific binding; they form stronger bonds to some DNA sequences than to others. Identification of a good binding site in the promoter for a gene suggests the possibility that the corresponding factor may play a role in the regulation of that gene. However, the sequences transcription factors recognize are typically short and allow for some amount of mismatch. Because of this, binding sites for a factor can typically be found at random every few hundred to a thousand base pairs. TESS has features to help sort through and evaluate the significance of predicted sites. PMID- 18428686 TI - The InterPro database and tools for protein domain analysis. AB - InterPro provides a one-stop shop for protein-sequence classification, freeing the user from having to visit multiple databases separately and rationalize the different results in varying formats. This unit describes how to submit a sequence to InterProScan via a Web server. It also provides instructions for installing and running InterProScan locally. In addition, details on browsing InterPro families and domains of interest using the InterPro Web and sequence retrieval system (SRS) are provided to show users how to get the most from the resource. PMID- 18428687 TI - Analysis of gene-expression data using J-Express. AB - The J-Express package has been designed to facilitate the analysis of microarray data with an emphasis on efficiency, usability, and comprehensibility. The J Express system provides a powerful and integrated platform for the analysis of microarray gene expression data. It is platform-independent in that it requires only the availability of a Java virtual machine on the system. The system includes a range of analysis tools and a project management system supporting the organization and documentation of an analysis project. This unit describes the J Express tool, emphasizing central concepts and principles, and gives examples of how it can be used to explore gene expression data sets. PMID- 18428688 TI - Installing a local copy of the Reactome Web site and database. AB - The Reactome project builds, maintains, and publishes a database of biological pathways. The information in the database is gathered from the experts in the field, peer reviewed, and edited by Reactome editorial staff and then published to the Reactome Web site, http://www.reactome.org (see UNIT ). Reactome software is open source and builds on top of other open-source or freely available software. Reactome data and code can be freely downloaded in its entirety and the Web site installed locally. This allows for more flexible interrogation of the data and also makes it possible to add one's own information to the database. PMID- 18428689 TI - Sequence file format conversion with command-line readseq. AB - One of the major challenges in using bioinformatics software is that there are a wide variety of sequence formats, e.g., GenBank, EMBL, and FASTA. It is often the case that a sequence or a set of sequences is in one format but is needed in another. This unit offers a solution to this problem--Readseq. Readseq is a program that can read and write 18 different formats. PMID- 18428690 TI - Using the TIGR gene index databases for biological discovery. AB - The TIGR Gene Index web pages provide access to analyses of ESTs and gene sequences for nearly 60 species, as well as a number of resources derived from these. Each species-specific database is presented using a common format with a homepage. A variety of methods exist that allow users to search each species specific database. Methods implemented currently include nucleotide or protein sequence queries using WU-BLAST, text-based searches using various sequence identifiers, searches by gene, tissue and library name, and searches using functional classes through Gene Ontology assignments. This protocol provides guidance for using the Gene Index Databases to extract information. PMID- 18428691 TI - Introduction to comparing large sequence sets. AB - Comparisons of whole genomes can yield important insights into the evolution of genome structure, such as the role of inversions in bacterial evolution and the identification of large-scale duplications in the human genome. This unit briefly compares two tools for aligning whole genome sequences: MUMmer and PipMaker. These tools differ in both the underlying algorithms used, and in the interface they present to the user. PMID- 18428692 TI - PipMaker: a World Wide Web server for genomic sequence alignments. AB - PipMaker is a World-Wide Web site used to compare two long genomic sequences and identify conserved segments between them. This unit describes the use of the PipMaker server and explains the resulting output files. PipMaker provides an efficient method of aligning genomic sequences and returns a compact, but easy-to interpret form of output, the percent identity plot (pip). For each aligning segment between two sequences the pip shows both the position relative to the first sequence and the degree of similarity. Optional annotations on the pip provide additional information to assist in the interpretation of the alignment. The default parameters of the underlying blastz alignment program are tuned for human-mouse alignments. PMID- 18428693 TI - Using MUMmer to identify similar regions in large sequence sets. AB - The MUMmer sequence alignment package is a suite of computer programs designed to detect regions of homology in long biological sequences. Version 2.1 makes several improvements to the package, including: increased speed and reduced memory requirements; the ability to handle both protein and DNA sequences; the ability to handle multiple sequence fragments; and new algorithms for clustering together basic matches. The system is particularly efficient at comparing highly similar sequences, such as alternative versions of fragment assemblies or closely related strains of the same bacterium. PMID- 18428694 TI - Large scale sequencing. AB - DNA sequencing and assembly is the Philosopher's Stone of bioinformatics, where the raw material of wet lab enzymatic chemistry is transformed into the bits and bytes of sequence information. The sequence information becomes the foundation for extensive downstream analyses. This unit provides a brief overview of the sequencing process. It describes several software programs typically used for base calling, assembling, and finishing sequence data such as STADEN, Phred, Phrap, and Consed. PMID- 18428695 TI - Viewing and editing assembled sequences using Consed. AB - Consed is a program for viewing and editing assemblies prepared by the Phrap assembly program. This unit presents a general introduction to Consed. It also provides instructions for identifying problem regions and searching in Consed. Additional protocols describe creating tags, picking primers, correcting joins and adding and removing reads. Instructions are also given for the use of Assembly View, predicting the results of a restriction digestion, the use of Consed in conjunction with PolyPhred, and using Autofinish, as well as a procedure for running Phred and Phrap. PMID- 18428696 TI - Identifying protein domains with the Pfam database. AB - Pfam is a database of such protein domain families, with each family represented by multiple sequence alignments and profile hidden Markov models (HMMs). In addition, each family has associated annotation, literature references and links to other databases. The entries in Pfam are available via the worldwide web and in flatfile format. This unit contains detailed information on how to access and utilise the information present in the Pfam database, namely the families, multiple alignments and annotation. Details on running Pfam, both remotely and locally are presented. PMID- 18428697 TI - Finding homologs in amino acid sequences using network BLAST searches. AB - BLAST, Basic Local Alignment Search Tool is used more frequently than any other biosequence database search program. The purpose of this unit is not only to show how to run searches on the Web, but also to demonstrate how to fine-tune arguments for a specific research project. It also offers guidance for interpreting results, handling statistical significance and biological relevance issues, and selecting complementary analyses. This unit covers three classes of the BLAST program: standard protein-to-protein searches, translated searches when either the query or the database consists of nucleotide sequences translated into proteins, and finally programs for comparing two sequences (as opposed to searching one sequence against a database of sequences). PMID- 18428698 TI - Constructing and refining multiple sequence alignments with PileUp, SeqLab, and the GCG suite. AB - This unit discusses how the Accelrys GCG Wisconsin Package SeqLab graphical user interface can be used to align, annotate, analyze, and export into alternative formats, multiple biological sequence data. The emphasis is on discovering and recognizing common elements within the dataset. The GCG programs, or implementations of public domain programs thereof, investigated include: LookUp, PileUp, PlotSimilarity, FASTA, Motifs, MEME/MotifSearch, the Profile Package, the HMMER Package, PAUPSearch, and ToFastA. ReadSeq, a non-GCG, public domain program is also used. PMID- 18428699 TI - An overview of multiple sequence alignment. AB - Multiple sequence alignment is perhaps the most commonly applied bioinformatics technique. It often leads to fundamental biological insight into sequence structure-function relationships of nucleotide or protein sequence families. In this unit, an overview of multiple sequence alignment techniques is presented, covering a history of nearly 30 years from the early pioneering methods to the current state-of-the-art techniques. Methodological and biological issues and end user considerations, as well as alignment evaluation issues, are discussed. PMID- 18428700 TI - Prokaryotic gene prediction using GeneMark and GeneMark.hmm. AB - In this unit, the GeneMark and GeneMark.hmm programs are presented as two different methods for the in silico prediction of genes in prokaryotes. GeneMark can be used for whole genome analysis as well as for the local analysis of a particular gene and its surrounding regions. GeneMark.hmm makes use of Hidden Markov models to find the transition points (boundaries) between protein coding states and noncoding states and can be efficiently used for larger genome sequences. These methods can be used in conjunction with each other for a higher sensitivity of gene detection. PMID- 18428701 TI - Eukaryotic gene prediction using GeneMark.hmm. AB - In this unit, eukaryotic GeneMark.hmm is presented as a method for detecting genes in eukaryotic DNA sequences. The eukaryotic GeneMark.hmm uses Markov models of protein coding and noncoding sequences, as well as positional nucleotide frequency matrices for prediction of the translational start, translational termination and splice sites. All these models along with length distributions of exons, introns and intergenic regions are integrated into one Hidden Markov model. The unit describes running the program over the Internet and locally on a Unix machine. It also discusses GeneMarkS EV, which can be used to detect genes in eukaryotic viruses. PMID- 18428702 TI - Application of FirstEF to find promoters and first exons in the human genome. AB - Predicting first exons and promoters is an important part of gene finding in DNA sequence analysis. This unit presents FirstEF as a method for predicting the first exons and promoters. A combines FirstEF predictions with other information such as cDNA/EST matches. PMID- 18428703 TI - Introduction to inferring evolutionary relationships. AB - This unit provides a general introduction to phylogeny. It defines common terms and discusses the issue of rooting trees, in addition to comparing gene and species trees. Methods for inferring phylogenies, such as distance methods, parsimony methods, and maximum likelihood are also presented. The unit concludes with discussion of how to assess tree confidence. PMID- 18428704 TI - Inferring evolutionary trees with PAUP*. AB - This unit provides a general description of reconstructing evolutionary trees using PAUP* 4.0. The protocol takes users through an example analysis of mitochondrial DNA sequence data using the parsimony and the likelihood criteria to infer optimal trees. The protocol also discusses searching options available in PAUP* and demonstrates how to import non-NEXUS formats. Finally, a general discussion is given regarding the pros and cons of the "model-free" and "model based" methods used throughout the protocol. PMID- 18428705 TI - Using MODELTEST and PAUP* to select a model of nucleotide substitution. AB - Models of nucleotide substitution are commonly used in the analysis of DNA sequences. This unit describes the use of the program MODELTEST (coupled with PAUP*) to find the best-fit model of substitution for the sequence alignment at hand. An example data file is analyzed and the interpretation of the results is discussed. Some background theory on model selection and a discussion of the relevance of models is included at the end of the unit. PMID- 18428706 TI - What if I don't have a tree? Split decomposition and related models. AB - A set of aligned character sequences or a matrix of evolutionary distances often contains a number of different and sometimes conflicting phylogenetic signals, and thus does not always support a unique tree. The method of split decomposition addresses this problem. For ideal data, this method gives rise to a phylogenetic tree, whereas less ideal data are represented by a tree-like network that may indicate evidence of different and conflicting phylogenies. The SplitsTree program, described here, implements this approach and can be used to compute and visualize phylogenetic networks called splits graphs. It also implements a number of distance transformations, the computation of parsimony splits, spectral analysis and bootstrapping. PMID- 18428707 TI - DRAGON and DRAGON view: information annotation and visualization tools for large scale expression data. AB - The Database Referencing of Array Genes ONline (DRAGON) database system consists of information derived from publicly available databases including UniGene, SWISS Prot, Pfam, and the Kyoto Encyclopedia of Genes and Genomes (KEGG). Through a Web accessible interface, the DRAGON Annotate tool rapidly supplies information pertaining to a range of biological characteristics of all the genes in any large scale gene expression data set. The subsequent inclusion of this information during data analysis and visualization allows for deeper insight into gene expression patterns. The set of DRAGON View tools provides methods for the analysis and visualization of expression patterns in relation to annotated information. Instead of incorporating the standard set of clustering and graphing tools available in many large-scale expression data analysis software packages, DRAGON View has been specifically designed to allow for the analysis of expression data in relation to the biological characteristics of gene sets. PMID- 18428708 TI - Analyzing molecular interactions. AB - This unit offers an overview of issues that must be considered when analyzing molecular interactions. The importance of bound solvent, binding affinity, and electrostatic interactions is discussed. PMID- 18428709 TI - Prediction of protein-protein interaction networks. AB - This unit offers a general overview of several techniques that have been developed for inferring functional and/or protein-protein interaction networks. The majority of these use whole-genome sequences as their primary input source of data. In addition, a few methods that utilize both protein features and experimental protein-protein interaction data directly in the prediction of new interactions have recently been developed. As a result, a slightly more detailed description of one such method is also provided. While not an exhaustive list of approaches, it is hoped that the reader will gain a sense of how these approaches are implemented, an idea of their relative strengths and weaknesses, as well as some perspective on the type of work being conducted in this highly active area of research. PMID- 18428710 TI - Evaluation of electrostatic interactions. AB - Described here are several computational procedures for the analysis of electrostatic interactions in molecular complexes, all based on a continuum model of solvation. The first section describes how to compute the residual potential, a measure of how electrostatically complementary a ligand is for its receptor. The second procedure describes electrostatic component analysis, a method by which the electrostatic contribution to the binding free energy can be broken up into terms directly attributable to individual chemical groups. Finally, electrostatic affinity optimization is described. This procedure is particularly useful in determining what portions of a ligand are the most suboptimal, and thus provide the greatest opportunity for the design of improvements. PMID- 18428711 TI - Using DelPhi to compute electrostatic potentials and assess their contribution to interactions. AB - There is a general agreement that electrostatic interactions play a significant role in the structure and function of biological molecules. However, obtaining quantitative estimation of the electrostatic energy requires computational models that capture the microscopic nature of the heterogeneous environment of macromolecules. This protocol offers elaboration on one of the common methods to calculate the electrostatic energetic contributions using continuum electrostatics. The method involves solving the Poisson-Boltzmann (PB) equation numerically and regarding the solute as having a homogenous dielectric constant. In order to apply this method, a three dimensional structure of the molecule derived from experimental data (crystallography, NMR) or modeling techniques is required. The protocol will focus on the DelPhi program (Accelrys Inc. San Diego), which is one of the most common programs used for the estimation of electrostatic free energy contribution. A simple procedure of assigning criteria and parameters (charge distribution, solvent and solute dielectric constants, iterations, grid resolution, etc) enables one to illustrate an electrostatic potential map and estimate the electrostatic free energy, although with limited accuracy. PMID- 18428712 TI - Searching the MINT database for protein interaction information. AB - The Molecular Interactions Database (MINT) is a relational database designed to store information about protein interactions. Expert curators extract the relevant information from the scientific literature and deposit it in a computer readable form. Currently (April 2003) MINT contains information on 4300 proteins and >3000 interactions. Although MINT contains interactions among proteins for several different species, the curators have initially focused on protein interactions in mammalian species. Protocols are provided here for searching MINT over the Internet, using the MINT Viewer. PMID- 18428713 TI - Structured Query Language (SQL) fundamentals. AB - Relational databases provide the most common platform for storing data. The Structured Query Language (SQL) is a powerful tool for interacting with relational database systems. SQL enables the user to concoct complex and powerful queries in a straightforward manner, allowing sophisticated data analysis using simple syntax and structure. This unit demonstrates how to use the MySQL package to build and interact with a relational database. PMID- 18428714 TI - Modeling biology using relational databases. AB - There are several different methodologies that can be used for designing a database schema; no one is the best for all occasions. This unit demonstrates two different techniques for designing relational tables and discusses when each should be used. These two techniques presented are (1) traditional Entity Relationship (E-R) modeling and (2) a hybrid method that combines aspects of data warehousing and E-R modeling. The method of choice depends on (1) how well the information and all its inherent relationships are understood, (2) what types of questions will be asked, (3) how many different types of data will be included, and (4) how much data exists. PMID- 18428715 TI - Searching the Mouse Genome Informatics (MGI) resources for information on mouse biology from genotype to phenotype. AB - The Mouse Genome Informatics (MGI, http://www.informatics.jax.org/) resource provides the research community with access to information on the genetics, genomics, and biology of the laboratory mouse. Core data in MGI include gene characterization and function, phenotype and disease model descriptions, DNA and protein sequence data, gene expression data, mammalian orthologies, polymorphisms, mapping data, and links to other bioinformatics databases. Semantic integration is supported through the use of standardized nomenclature and controlled vocabularies such as the mouse Anatomical Dictionary and the Gene Ontologies. MouseBLAST offers a fast, sequence-based method for identifying genes and searching for similarity, and provides links from sequence to MGI biological data. MGI extracts and organizes data from primary literature. MGI data are shared with and widely displayed by other bioinformatics resources. The database is updated nightly with curated annotations and regularly adds new datasets and enhances user interface features. This unit provides a guide to using the MGI bioinformatics resource. PMID- 18428716 TI - RNA secondary structure analysis using the Vienna RNA package. AB - This Current Protocols in Bioinformatics unit documents how to use the Vienna RNA package for RNA secondary structure analysis. Possible tasks include structure prediction for single sequences, prediction of consensus structures, and sequence design. PMID- 18428717 TI - RNAi: design and analysis. AB - RNAi is a novel genetic tool that is poised to revolutionize the study of genetics and gene networks. Use of this tool requires knowledge of the underlying mechanisms and application of this knowledge to the construction of a gene silencing library. This unit discusses approaches to rational design of this library and its use in experiments. It focuses on the design algorithms developed so far and briefly discuss several issues that could lead to improvements in the current design algorithms. PMID- 18428718 TI - Predicting the secondary structure common to two RNA sequences with Dynalign. AB - Dynalign is a dynamic programming algorithm for the simultaneous prediction of the lowest-free-energy secondary structure common to two RNA sequences and the alignment of the two sequences. It has been shown that the average accuracy of secondary structure prediction is improved using Dynalign, as compared to free energy minimization of a single sequence. This unit provides protocols for using Dynalign on a Microsoft Windows platform as part of the RNAstructure package, and for compiling and using Dynalign on Unix/Linux computers. PMID- 18428719 TI - Finding protein sequences using PROWL. AB - PROWL is a collection of tools for the identification of protein sequences, using input data derived from mass spectrometry. Experimental data from various types of mass spectrometers can be input directly into PROWL's component software. This unit presents protocols for several of the individual PROWL tools. Specifically, PepFrag allows for the analysis of a single spectrum derived from tandem mass spectrometry. GPM, on the other hand, provides for the analysis of multiple MS/MS spectra. An additional protocol introduces ProFound for analyzing a single spectrum of peptide mass fingerprinting data. PMID- 18428720 TI - Mathematically complete nucleotide and protein sequence searching using Ssearch. AB - In this unit a protocol is described for predicting the structure of simple transmembrane a-helical bundles. The protocol is based on a global molecular dynamics search (GMDS) of the configuration space of the helical bundle, yielding several candidate structures. The correct structure among these candidates is selected using information from silent amino acid substitutions, employing the premise that only the correct structure must (by definition) accept all of the silent amino acid substitutions. Thus, the correct structure is found by repeating the GMDS for several close homologs and selecting the structure that persists in all of the trials. PMID- 18428721 TI - Installing, maintaining, and using a local copy of BLAST for intranet and workstation use. AB - The Basic Local Alignment Search Tool (BLAST) is one of the widest used and most useful applications in sequence-based bioinformatics analysis. Frequently it is not practical or possible to use remote BLAST services through the Internet due to restrictions of a security or technical nature or the need for high-throughput analysis requiring greater amounts of processing power than are available from remote services. This unit describes the steps involved in obtaining and installing a copy of the BLAST software for use on a local intranet or stand alone workstation. Once installed, the BLAST package can be used to create BLAST searchable nucleotide and protein sequence databanks. Various popular hardware (PPC, Intel) and operating system (MacOSX, FreeBSD and Linux) options for running and maintaining the software are discussed. Finally, steps for indexing proprietary and third party (publicly available) sequence databanks for use with BLAST and managing these resources are discussed. PMID- 18428722 TI - Computing multiple sequence/structure alignments with the T-coffee package. AB - The FASTA package provides a comprehensive set of similarity searching programs, similar to those provided by the BLAST package, and some additional programs for searching with short peptides and oligonucleotides that are not provided by BLAST. The FASTA programs work with a wide variety of database formats, including mySQL sequence databases. FASTA provides very accurate statistical significance estimates, and is more sensitive than BLASTN when comparing DNA sequences. These protocols describe how to use the FASTA programs to characterize protein and DNA sequences, using protein:protein, protein:DNA, and DNA:DNA comparisons. PMID- 18428723 TI - Finding protein and nucleotide similarities with FASTA. AB - The FASTA package provides a comprehensive set of similarity searching programs, similar to those provided by the BLAST package, and some additional programs that are not provided by BLAST for searching with short peptides and oligonucleotides. The FASTA programs work with a wide variety of database formats, including mySQL sequence databases. FASTA provides very accurate statistical significance estimates, and is more sensitive than BLASTN when comparing DNA sequences. These protocols describe how to use the FASTA programs to characterize protein and DNA sequences, using protein:protein, protein:DNA, and DNA:DNA comparisons. PMID- 18428724 TI - An overview of gene identification: approaches, strategies, and considerations. AB - Modern biology is on the verge of officially ushering in a new era in science with the completion of the sequencing of the human genome in April 2003. While often erroneously called the "post-genome era", this will actually truly mark the beginning of the "genome era," a time in which the availability of sequence data for many genomes will have a significant effect on how science is performed in the 21st century. This unit offers an overview of many of the gene prediction methods that are currently available and offers a general assessment of how well the methods work for various problems. PMID- 18428725 TI - Using RepeatMasker to identify repetitive elements in genomic sequences. AB - The RepeatMasker program is used for identifying repetitive elements in nucleotide sequences for further detailed analyses. Users can run RepeatMasker remotely via a Web site, or, for larger input sequences, the program and its dependent programs may be downloaded and run locally on Unix/Linux computers. The protocols in this chapter detail how to use RepeatMasker both remotely and locally to extract repetitive elements data and mask these repetitive elements in nucleotide sequences. PMID- 18428726 TI - GrailEXP and Genome Analysis Pipeline for genome annotation. AB - The Basic Protocol describes the use of GrailEXP, the latest version of the gene finding system from Oak Ridge National Laboratory. GrailEXP provides gene models, by making use of sequence similarity with Expressed Sequence Tags (ESTs) and known genes. GrailEXP also provides alternatively spliced constructs for each gene based on the available EST evidence. The Support Protocol describes the use of the Genome Analysis Pipeline, a web application which allows users to perform comprehensive sequence analysis by offering a selection from a wide choice of supported gene finders, other biological feature finders, and database searches. PMID- 18428727 TI - FAMS and FAMSBASE for protein structure. AB - The computer program FAMS (Full Automatic Modeling System) performs homology modeling of protein structures by means of an algorithm consisting of database searches and simulated annealing. FAMS produces a model in which the torsion angles of the backbone and sidechains are highly accurate. The operations of the FAMS algorithm are fully automated, and, therefore, special knowledge, techniques or experience are not required in order to obtain a biologically worthwhile protein structure. PMID- 18428728 TI - Modeling membrane proteins utilizing information from silent amino acid substitutions. AB - This unit describes predicting the structure of simple transmembrane alpha helical bundles. The protocol is based on a global molecular dynamics search (GMDS) of the configuration space of the helical bundle, yielding several candidates structures. The correct structure amongst these candidates is selected using information from silent amino acid substitutions, employing the following premise: Only the correct structure must (by definition) accept all of the silent amino acid substitutions. Thus, the correct structure is found by repeating the GMDS for several close homologues and selecting the structure that persists in all of the trials. PMID- 18428729 TI - Using PEBBLE for the evolutionary analysis of serially sampled molecular sequences. AB - The PEBBLE (Phylogenetics, Evolutionary Biology, and Bioinformatics in a moduLar Environment) application is a relative newcomer to the field of phylogenetic applications. Although designed as a customizable generalist application, PEBBLE was initially developed to implement procedures for the analysis of sequences associated with different sampling times, e.g., rapidly evolving viral genes sampled over the course of infection, or ancient DNA sequences. The basic protocol describes the use of PEBBLE to infer a phylogenetic tree using the sUPGMA algorithm, and the inference of substitution rate parameters using maximum likelihood. The alternate and support protocols describe the simulation capabilities of PEBBLE, and general use of the PEBBLE application, respectively. PMID- 18428730 TI - Microarray data visualization and analysis with the Longhorn Array Database (LAD). AB - The wide availability and use of DNA microarrays has brought the power of whole genome functional characterization to a large variety of research environments. Microarrays, however, also introduce significant infrastructural and analytical concerns with respect to the long-term warehousing, annotation, and visualization of immense data sets. The Longhorn Array Database (LAD) is a MIAME-compliant microarray database that operates on PostgreSQL and Linux. It is a free and completely open-source software solution, and provides a systematic and proven environment in which vast experiment sets can be safely archived, securely accessed, biologically annotated, quantitatively analyzed, and visually explored. This unit provides the complete set of information needed to successfully deploy, configure, and use LAD for the purposes of two-color DNA microarray analysis and visualization. PMID- 18428731 TI - Using GenMAPP and MAPPFinder to view microarray data on biological pathways and identify global trends in the data. AB - GenMAPP (Gene MicroArray Pathway Profiler) is a free, stand-alone computer program designed for viewing and analyzing gene expression data on MAPPs representing biological pathways or any other functional grouping of genes. A MAPP is a special file format produced with the graphics tools in GenMAPP that depicts the biological relationship between genes or gene products. When a MAPP is linked to an expression dataset, GenMAPP automatically and dynamically color codes the genes on the MAPP according to criteria supplied by the user. MAPPFinder is an accessory program that works with GenMAPP and the annotations from the Gene Ontology (GO) Consortium to identify global biological trends in gene expression data. MAPPFinder relates the microarray dataset to the GO hierarchy and calculates a percentage and statistical score for genes meeting the user's criterion for a meaningful gene expression change for each GO biological process, cellular component, and molecular function term. PMID- 18428732 TI - Integrating whole-genome expression results into metabolic networks with Pathway Processor. AB - Genes never act alone in a biological system, but participate in a cascade of networks. As a result, analyzing microarray data from a pathway perspective leads to a new level of understanding the system. The authors' group has recently developed Pathway Processor (http://cgr.harvard.edu/cavalieri/pp.html), an automatic statistical method to determine which pathways are most affected by transcriptional changes and to map expression data from multiple whole-genome expression experiments on metabolic pathways. This unit presents applications of the Pathway Processor software. PMID- 18428733 TI - An overview of Spotfire for gene-expression studies. AB - Spotfire DecisionSite for Functional Genomics (referred to here as Spotfire) is a powerful data mining and visualization application with use in many disciplines. This unit provides an overview of Spotfire's utility in analyzing gene expression data obtained from DNA microarray experiments. Analysis of microarray data requires software-based solutions able to handle and manipulate the enormous amount of data generated. Spotfire provides a solution for accessing, analyzing and visualizing data generated from microarray experiments. Spotfire is designed to allow biologists with little or no programming or statistical skills to transform, process, and analyze microarray data. PMID- 18428734 TI - Loading and preparing data for analysis in spotfire. AB - This unit strictly focuses on data preparation within Spotfire. Microarray data exist in a variety of formats, which often depend on the particular array technology and detection instruments used. The first protocols in this unit describe loading Affymetrix and GenePix data into Spotfire. Once the data are loaded, it is necessary to filter and preprocess the data prior to analysis. Subsequently, the data transformation and normalization techniques presented here, are critical to correctly performing powerful microarray data mining expeditions. These steps extract or enhance meaningful data characteristics and prepare the data for the application of certain analysis methods such as statistical tests to compute significance and clustering methods-which mostly require data to be normally distributed. The unit outlines several methods for normalizing the data within an experiment and between multiple experiments. PMID- 18428735 TI - Analyzing and visualizing expression data with Spotfire. AB - This unit assumes the reader is familiar with the Spotfire environment, has successfully installed Spotfire, and has uploaded and prepared data for analysis. It presents numerous methods for analyzing microarray data. Specifically, the first two protocols describe methods for identifying differentially expressed genes via the t-test/ANOVA and the distinction calculation respectively. Another protocol discusses how to conduct a profile search. Additional protocols illustrate various clustering methods, such as hierarchical clustering, K-means clustering, and principal components analysis. A protocol explaining coincidence testing allows the reader to compare the results from multiple clustering methods. Additional protocols demonstrate querying the Internet for information based on the microarray data, mathematically transforming data within Spotfire to generate new data columns, and exporting Spotfire visualizations. PMID- 18428736 TI - Identifying functional sites based on prediction of charged group behavior. AB - This protocol describes the implementation and interpretation of THEMATICS, a simple computational predictor of functional information for proteins from the three-dimensional structure. This method is based on the computation of the electrical potential function for the protein and the calculation of the predicted titration curves for each of the titratable groups in the protein. While most of the titratable residues in a protein have predicted titration behavior that fits the Henderson-Hasselbalch equation, the ionizable residues in the active site generally deviate dramatically from the typical behavior. From the calculated titration curves, one identifies those residues that deviate significantly from Henderson-Hasselbalch behavior. A cluster of two or more of such deviant titratable residues in physical proximity is a reliable predictor of active-site location. PMID- 18428737 TI - Using the Reactome database. AB - There is considerable interest in the bioinformatics community in creating pathway databases. The Reactome project (a collaboration between Cold Spring Harbor Laboratory and the European Bioinformatics Institute) is one such pathway database and collects structured information on all the biological pathways in the human. It is a curated collection of well-documented molecular reactions that span the gamut from simple intermediate metabolism to complex cellular events. This information is supplemented with likely orthologous molecular reactions in mouse, rat, zebrafish, and other model organisms. This unit describes how to use the Reactome database to learn the steps of a biological pathway and see how one pathway interacts with another; navigate and browse through the Reactome database; identify the pathways in which a molecule of interest is involved; use the Pathfinder tool to search the database for possible connections within and between pathways. PMID- 18428738 TI - Analyzing networks with VisANT. AB - The VisANT tool, accessible from any recent Java-enabled browser, is a platform independent, flexible, Web-enabled program for quick and simple construction, visualization, and analysis of molecular and higher order networks based on functional (e.g., expression profiles, phylogenetic profiles) and physical (e.g., yeast two-hybrid, chromatin-immunoprecipitation) relations from either the Predictome database or user-defined data sets. Analysis capabilities include identification of feed-forward and -back loops, shortest paths, and node degree distribution. Additionally, network constructs can be saved, accessed, and shared online. VisANT is able to develop and display meta-networks for meta-nodes that are structural complexes or pathways (soon including nodes representing any kind of dense cluster). Further, VisANT supports a growing number of standard exchange formats and database referencing standards, e.g., KEGG/KGML, BioPAX (in progress), GenBank, Gene Ontology. Multiple species are supported to the extent that computed or experimental evidence of interactions or associations are available (i.e., public datasets or Predictome database). PMID- 18428739 TI - Using relational databases for improved sequence similarity searching and large scale genomic analyses. AB - Relational databases are designed to integrate diverse types of information and manage large sets of search results, greatly simplifying genome-scale analyses. Relational databases are essential for management and analysis of large-scale sequence analyses, and can also be used to improve the statistical significance of similarity searches by focusing on subsets of sequence libraries most likely to contain homologs. This unit describes using relational databases to improve the efficiency of sequence similarity searching and to demonstrate various large scale genomic analyses of homology-related data. This unit describes the installation and use of a simple protein sequence database, seqdb_demo, which is used as a basis for the other protocols. These include basic use of the database to generate a novel sequence library subset, how to extend and use seqdb_demo for the storage of sequence similarity search results and making use of various kinds of stored search results to address aspects of comparative genomic analysis. PMID- 18428740 TI - Human mutation databases. AB - The first part of this unit compares general and locus-specific mutation databases. The second section deals with submitting data. The third part provides guidance for accessing mutation data. The final section offers advice on database construction. PMID- 18428741 TI - Using the Arabidopsis Information Resource (TAIR) to find information about Arabidopsis genes. AB - The Arabidopsis Information Resource (TAIR; http://www.arabidopsis.org) is a comprehensive Web resource of Arabidopsis biology for plant scientists. TAIR curates and integrates information about genes, proteins, gene expression, mutant phenotypes, biological materials such as DNA and seed stocks, genetic markers, genetic and physical maps, biochemical pathways, genome organization, images of mutant plants and protein sub-cellular localizations, publications, and the research community Data in TAIR are extensively interconnected and can be accessed through a variety of Web-based search and display tools. This unit primarily focuses on some basic methods for searching, browsing, visualizing, and analyzing information about Arabidopsis genes. Gene expression data from microarrays is a recent addition to the database and methods for accessing these data are also described. Two pattern identification programs are described for mining TAIR's unique Arabidopsis sequence data sets. We also describe how to use AraCyc for mining plant metabolic pathways. PMID- 18428742 TI - Using the KEGG database resource. AB - KEGG (Kyoto Encyclopedia of Genes and Genomes) is a bioinformatics resource for understanding the functions and utilities of cells and organisms from both high level and genomic perspectives. It is a self-sufficient, integrated resource consisting of genomic, chemical, and network information, with cross-references to numerous outside databases, containing a complete set of building blocks (genes and molecules) and wiring diagrams (interaction networks) to represent cellular functions. This unit describes protocols for using KEGG, KEGG PATHWAY, KEGG GENES, KEGG SSDB, KEGG EXPRESSION, and KEGG LIGAND. Protocols are also described for how to color maps, compare chemical compounds and glycan chains, analyze Ortholog Clusters, and visualize and analyze microarray data, among other procedures. All of these many protocols enable the user to take advantage of the full breadth of the functionality provided by KEGG. PMID- 18428743 TI - MultiPipMaker: comparative alignment server for multiple DNA sequences. AB - The MultiPipMaker World Wide Web server (http://www.bx.psu.edu) provides a useful tool for aligning multiple sequences and visualizing regions of conservation between them. This unit describes the use of the MultiPipMaker server and gives an explanation of the resulting output files and supporting tools. Features provided by the server include alignment of up to 20 very long genomic sequences, output choices of a true, nucleotide-level multiple alignment or stacked, pairwise percent identity plots, and user-specified annotations for genomic features and elements of choice, with clickable links to additional information. Alignments can include unordered, unoriented secondary sequences. PMID- 18428744 TI - Generating a genome assembly with PCAP. AB - This unit describes how to use the Parallel Contig Assembly Program (PCAP) to assemble the data produced by a whole-genome shotgun sequencing project. We present a basic protocol for using PCAP on a multiprocessor computer in a 300-Mb genome assembly project. A support protocol to prepare input files for PCAP is also described. Another basic protocol for using PCAP on a distributed cluster of computers in a 3-Gb genome assembly project is presented, in addition to suggestions for understanding results from PCAP. PMID- 18428745 TI - Annotating non-coding RNAs with Rfam. AB - Non-coding RNA (ncRNA) genes produce a functional RNA product, rather than a translated protein. The range and importance of such genes is only recently apparent, with known ncRNAs participating in a wide range of structural, regulatory, and catalytic roles within the cell. Like protein-coding genes, multiple sequence alignments of families of ncRNAs tell us much about their structure and function, and enable the formulation of statistical models for the detection of related sequences. Rfam is a database of families of ncRNAs, represented by structure-annotated multiple sequence alignments and covariance models. PMID- 18428747 TI - Protein identification using TurboSEQUEST. AB - SEQUEST is the most widely used software tool for identifying proteins in complex mixtures. It is a mature, robust program that identifies peptides directly from uninterpreted tandem mass spectra, thus making large-scale proteomic studies possible. Thermo Electron's TurboSEQUEST provides a Windows-based graphical user interface for running SEQUEST and interpreting results. The protocol in this unit describes the basic steps involved in processing mass spectrometric data and analyzing results using TurboSEQUEST. It also provides an overview of the SEQUEST algorithm and a discussion of data filtering methods, critical issues in data interpretation, and available resources that can facilitate proper interpretation of SEQUEST results. PMID- 18428748 TI - Using the Gibbs motif sampler to find conserved domains in DNA and protein sequences. AB - The Gibbs Motif Sampler (Gibbs) is a software package for discovering conserved elements in biopolymer sequences. This unit describes the basic operation of the Web-based interface to Gibbs, along with advanced examples of its use, and the Web interface to dscan, a sequence database search program. PMID- 18428749 TI - Using CorePromoter to find human core promoters. AB - The CorePromoter program is very useful for identification of transcriptional start sites (TSS) and core promoter regions when 5'-upstream genomic DNA sequences of human genes are available. It is very simple to use and can be accessed either through the Web or after downloading to a local computer. The protocols in this unit introduce its basic methodology and discuss how to apply it to a sample problem in conjunction with other gene-finding programs. PMID- 18428746 TI - Proteomics and the analysis of proteomic data: an overview of current protein profiling technologies. AB - In recent years, several proteomic methodologies have been developed that now make it possible to identify, characterize, and comparatively quantify the relative level of expression of hundreds of proteins that are coexpressed in a given cell type or tissue, or that are found in biological fluids such as serum. These advances have resulted from the integration of diverse scientific disciplines including molecular and cellular biology, protein/peptide chemistry, bioinformatics, analytical and bioanalytical chemistry, and the use of instrumental and software tools such as multidimensional electrophoretic and chromatographic separations and mass spectrometry. In this unit, some of the common protein-profiling technologies are reviewed, along with the accompanying data-analysis tools. PMID- 18428750 TI - Representing structural information with RasMol. AB - This unit describes the use of RasMol, a popular interactive molecular graphics program, for the display of biomolecular structures. The unit begins with a general guide to obtaining molecular coordinates from the Protein Data Bank and displaying them with RasMol, and then describes several molecular representations that are useful for different applications, along with methods for ensuring that one is viewing the appropriate biological unit and some ideas for customizing one's molecular graphics session. PMID- 18428751 TI - Phylogenomic inference of protein molecular function. AB - With the explosion in sequence data, accurate prediction of protein function has become a vital task in prioritizing experimental investigation. While computationally efficient methods for homology-based function prediction have been developed to make this approach feasible in high-throughput mode, it is not without its dangers. Biological processes such as gene duplication, domain shuffling, and speciation produce families of related genes whose gene products can have vastly different molecular functions. Standard sequence-comparison approaches may not discriminate effectively among these candidate homologs, leading to errors in database annotations. In this unit, we describe phylogenomic approaches to reduce the error rate in function prediction. Phylogenomic inference of protein molecular function consists of a series of subtasks. Once a cluster of homologs is identified, a multiple sequence alignment and phylogenetic tree are constructed. Finally, the phylogenetic tree is overlaid with experimental data culled for the members of the family, and changes in biochemical function can be traced along the evolutionary tree. PMID- 18428752 TI - Gene expression analysis via multidimensional scaling. AB - Expression profiling of biological samples using microarray technologies has proven to be a powerful tool for molecular classification and biomarker identification. Visualization of similarities between biological samples from their molecular signatures is essential in forming new hypotheses. Multidimensional scaling is one of the methods that converts the structure in the similarity matrix to a simple geometrical picture: the larger the dissimilarity between two samples (evaluated through gene expression profiling), the further apart the points representing the experiments in the picture should be. In this unit, we will discuss the mathematical fundamentals of this method, along with step-by-step procedures that allow users to quickly obtain the results, provided that all necessary resources are ready. Examples of applying the MDS program and the interpretation of these results are also provided in this unit. PMID- 18428753 TI - The importance of biological databases in biological discovery. AB - Biological databases play a central role in bioinformatics. They offer scientists the opportunity to access sequence and structure data for tens of thousands of sequences from a broad range of organisms. This unit provides a brief overview of major sequence databases and portals, such as GenBank, UCSC Genome Browser, and Ensembl. Model organism databases, including the TIGR Gene Indices, WormBase, and the Arabidopsis Information Resource (TAIR) are also covered. Non-sequence centric databases such as Online Mendelian Inheritance in Man (OMIM), the Protein Data Bank (PDB), and the Kyoto Encyclopedia of Genes and Genomes (KEGG) are also discussed. PMID- 18428754 TI - The Human Gene Mutation Database (HGMD) and its exploitation in the study of mutational mechanisms. AB - The Human Gene Mutation Database (HGMD) constitutes a comprehensive core collection of data on germ-line mutations in nuclear genes underlying or associated with human inherited disease (http://www.hgmd.org). Data cataloged include single base-pair substitutions in coding, regulatory, and splicing relevant regions, microdeletions and microinsertions, indels, and triplet repeat expansions, as well as gross gene deletions, insertions, duplications, and complex rearrangements. Each mutation is entered into HGMD only once, in order to avoid confusion between recurrent and identical-by-descent lesions. By June 2005, the database contained in excess of 53,000 different lesions detected in 2029 different nuclear genes, with new entries currently accumulating at a rate in excess of 5000 per annum. HGMD includes cDNA reference sequences, now provided for more than 90% of the listed genes, splice junction data, disease-associated and functional polymorphisms, and links to data present in publicly available online locus-specific mutation databases. PMID- 18428755 TI - Exploring phenotypic data at the rat genome database. AB - The laboratory rat, Rattus norvegicus, is an important model of human health and disease, and experimental findings in the rat have direct relevance to human based research. The Rat Genome Database (RGD, http://rgd.mcw.edu) is a model organism database that provides access to wide variety of curated rat data such as genes and their homologs, quantitative trait loci, phenotypes, comparative mapping, and genome analysis. We present an overview of the database followed by specific examples that can be used to gain experience in employing RGD to explore the wealth of functional data available for the rat. We show how to make associations with the genome and use comparative tools to link the rat with human and mouse in order to integrate results from these three species of critical biomedical importance. PMID- 18428756 TI - Searching the NCBI databases using Entrez. AB - One of the most widely-used interfaces for the retrieval of information from biological databases is the NCBI Entrez system. Entrez capitalizes on the fact that there are pre-existing, logical relationships between the individual entries found in numerous public databases. The existence of such natural connections, mostly biological in nature, argued for the development of a method through which all the information about a particular biological entity could be found without having to sequentially visit and query disparate databases. Two Basic Protocols describe simple, text-based searches, illustrating the types of information that can be retrieved through the Entrez system. An Alternate Protocol builds upon the first Basic Protocol, using additional, built-in features of the Entrez system, and providing alternative ways to issue the initial query. The Support Protocol reviews how to save frequently-issued queries. Finally, Cn3D, a structure visualization tool, is also discussed. PMID- 18428757 TI - Searching WormBase for information about Caenorhabditis elegans. AB - WormBase is the major public biological database for the nematode Caenorhabditis elegans. It is meant to be useful to any biologist who wants to use C. elegans, whatever his or her specialty. WormBase contains information about the genomic sequence of C. elegans, its genes and their products, and its higher-level traits such as gene expression patterns and neuronal connectivity. WormBase also contains genomic sequences and gene structures of C. briggsae and C. remanei, two closely related worms. These data are interconnected, so that a search beginning with one object (such as a gene) can be directed to related objects of a different type (e.g., the DNA sequence of the gene or the cells in which the gene is active). One can also perform searches for complex data sets. The WormBase developers group actively invites suggestions for improvements from the database users. WormBase's source code and underlying database are freely available for local installation and modification. PMID- 18428758 TI - An overview of RNA structure prediction and applications to RNA gene prediction and RNAi design. AB - This unit briefly describes the two fundamentally different methods for predicting RNA structures. The first is to find that structure with the minimum free energy of folding, as predicted by various thermodynamic parameters related to base-pair stacking, loop lengths, and other features. If one has only a single sequence, this thermodynamic approach is the best available method. The second fundamental approach to RNA structure prediction is to use multiple, homologous sequences for which one can infer a common structure, and then try and predict a structure common to all of the sequences. Such an approach is referred to as a comparative method or phylogenetic method of RNA structure prediction. PMID- 18428759 TI - RNA secondary structure analysis using RNAstructure. AB - RNAstructure is a user-friendly program for the prediction and analysis of RNA secondary structure under Microsoft Windows. This unit provides protocols for RNA secondary structure prediction and prediction of high-affinity oligonucleotide binding sites to a structured RNA target. PMID- 18428760 TI - Using Pharmabase to perform pharmacological analyses of cell function. AB - Pharmabase is designed to form a bridge between the molecular dimension of cell transport processes and the functional manipulation of the protein players. It has as its emphasis membrane transport and related pharmacology. Several search and navigation options are available, including membrane transport, disease, and a graphic interface arranged by pathway and cell type. The level of entry to the database can be tailored to the investigator's level of expertise. The final product of Pharmabase is a Compound Record detailing the use and targets of individual compounds. Navigation routes generally fall into hierarchical keys and are cross-referenced. Pharmabase encourages input from its user community. It is maintained by the BioCurrents Research Center, an NIH resource funded through the National Center for Research Resources (NCRR). PMID- 18428761 TI - Using MSDchem to search the PDB ligand dictionary. AB - The PDB ligand dictionary is the chemical reference database of all the small building block molecules (e.g., amino acids, nucleic acids, and bound ligands) in the Protein Data Bank (PDB) referenced by a distinct three-letter code identifier. Since PDB files have only three-dimensional coordinate data, the role of the dictionary that of a reference resource for the actual chemical properties of small molecules, shared consistently across all PDB entries. The ligand dictionary is maintained in all sites of the Worldwide Protein Data Bank (wwPDB), the Research Collaboratory for Structural Bioinformatics (RCSB) in U.S., the Macromolecular Structure Database (MSD) in Europe, and the Protein Data Bank in Japan (PDBj), and it is exchanged on a regular basis. The MSD group at the European BioInformatics Institute (EBI) extends the dictionary into the MSDchem ligand database, which utilizes chemo-informatics packages and incorporates additional curation work. MSDchem is publicly available on the Web through the MSDchem search system, the functionality of which is described in more detail in this unit. PMID- 18428762 TI - An introduction to recognizing functional domains. AB - This unit provides an overview of issues involved in domain recognition in protein and DNA sequences. It opens with a discussion of the two primary methods of domain representation, namely consensus sequences and alignment matrices (e.g., the log-odds matrix). The unit continues with a brief overview of some of the resources available for identifying functional domains in nucleotide sequences (e.g., TRANSFAC). In addition, it reviews databases such as Pfam, InterPro and Blocks, which are available for protein analysis. PMID- 18428763 TI - Using the Structure-function Linkage Database to characterize functional domains in enzymes. AB - The Structure-Function Linkage Database (SFLD; http://sfld.rbvi.ucsf.edu/) is a Web-accessible database designed to link enzyme sequence, structure, and functional information. This unit describes the protocols by which a user may query the database to predict the function of newly sequenced enzymes and to correct misannotated functional assignments for enzymes currently in public databases. It is especially useful in helping a user discriminate functional capabilities of a sequence that is only distantly related to characterized sequences in publicly available databases. PMID- 18428764 TI - Using Weeder for the discovery of conserved transcription factor binding sites. AB - One of the greatest challenges facing modern molecular biology is the understanding of the complex mechanisms regulating gene expression. A fundamental step in this process requires the characterization of motifs involved in the regulation of gene expression at transcriptional and post-transcriptional levels. In particular, transcription is modulated by the interaction of transcription factors with their corresponding binding sites. Weeder is a software package freely available for non commercial users as a stand-alone or Web-based application for the automatic discovery of conserved motifs in a set of related DNA sequences from coregulated genes. The motifs found are likely to represent instances of binding sites for some common transcription factor regulating the genes of the set. The program has been designed to make its usage as simple as possible and to require very little prior knowledge about the length and conservation of the motifs to be found. PMID- 18428765 TI - An introduction to modeling structure from sequence. AB - The underlying premise behind all attempts to determine a large number of diverse protein structures is that the total number of protein domain folds is much smaller, by many orders of magnitude, than the total number of sequences; in other words, many sequences adopt essentially the same fold. If the fold of a protein could be recognized from sequence information alone, then a complete database of all possible folds would allow the structure corresponding to any sequence to be modeled. The growth of structure determination has turned most biochemists and biologists into consumers of structural information. As the demand for such information continues to outstrip the supply, all aspects of structure modeling assume increasing importance. This unit provides an introduction to modeling structure from its sequence and surveys the currently available methods described in the subsequent units of this chapter. PMID- 18428766 TI - Using Dali for structural comparison of proteins. AB - The Dali program is widely used for carrying out automatic comparisons of protein structures determined by X-ray crystallography or NMR. The most familiar version is the Dali server, which performs a database search comparing a query structure supplied by the user against the database of known structures (PDB) and returns the list of structural neighbors by e-mail. The more recently introduced DaliLite server compares two structures against each other and visualizes the result interactively. The Dali database is a structural classification based on precomputed all-against-all structural similarities within the PDB. The resulting hierarchical classification can be browsed on the Web and is linked to protein sequence classification resources. All Dali resources use an identical algorithm for structure comparison. Users may run Dali using the Web, or the program may be downloaded to be run locally on Linux computers. PMID- 18428768 TI - Getting a tree fast: Neighbor Joining, FastME, and distance-based methods. AB - Neighbor Joining (NJ), FastME, and other distance-based programs including BIONJ, WEIGHBOR, and (to some extent) FITCH, are fast methods to build phylogenetic trees. This makes them particularly effective for large-scale studies or for bootstrap analysis, which require runs on multiple data sets. Like maximum likelihood methods, distance methods are based on a sequence evolution model that is used to estimate the matrix of pairwise evolutionary distances. Computer simulations indicate that the topological accuracy of FastME is best, followed by FITCH, WEIGHBOR, and BIONJ, while NJ is worse. Moreover, FastME is even faster than NJ with large data sets. Best-distance methods are equivalent to parsimony in most cases, but become more accurate when the molecular clock is strongly violated or in the presence of long (e.g., outgroup) branches. This unit describes how to use distance-based methods, focusing on NJ (the most popular) and FastME (the most efficient today). It also describes how to estimate evolutionary distances from DNA and proteins, how to perform bootstrap analysis, and how to use CLUSTAL to compute both a sequence alignment and a phylogenetic tree. PMID- 18428769 TI - Active site profiling to identify protein functional sites in sequences and structures using the Deacon Active Site Profiler (DASP). AB - Methods for the annotation and analysis of functional sites in proteins are an area of active research, and those methods that allow detailed characterization of functional site features are much needed. A Web site application, DASP, which implements a previously described method (Cammer, et al., 2003) to allow users to create an active site profile for any protein family, is described. Two protocols for functional site analysis of protein families using DASP are presented: 1) creation of functional site signatures and a profile from proteins of known structure and 2) utilization of the active site profile to search sequences that contain fragments similar to those found in the functional site signatures. The active site profile produced by Basic Protocol 1 allows the user to analyze the features of the functional site, i.e., those characteristics that are common across the family and those that are unique to one or several members of the family. The characteristics that are unique to a subfamily might be described as specificity determinants i.e., features that impart specificity to a particular function. Basic Protocol 2 provides instructions for searching for sequences that might contain a similar functional site. PMID- 18428771 TI - Using Apollo to browse and edit genome annotations. AB - An annotation is any feature that can be tied to genomic sequence, such as an exon, transcript, promoter, or transposable element. As biological knowledge increases, annotations of different types need to be added and modified, and links to other sources of information need to be incorporated, to allow biologists to easily access all of the available sequence analysis data and design appropriate experiments. The Apollo genome browser and editor offers biologists these capabilities. Apollo can display many different types of computational evidence, such as alignments and similarities based on BLAST searches (UNITS 3.3 & 3.4), and enables biologists to utilize computational evidence to create and edit gene models and other genomic features, e.g., using experimental evidence to refine exon-intron structures predicted by gene prediction algorithms. This protocol describes simple ways to browse genome annotation data, as well as techniques for editing annotations and loading data from different sources. PMID- 18428770 TI - Searching, viewing, and visualizing data in the Biomolecular Interaction Network Database (BIND). AB - The Biomolecular Interaction Network Database (BIND) comprises data from peer reviewed literature and direct submissions. BIND's data model was the first of its kind to be peer-reviewed prior to database development, and is now a mature standard data format spanning molecular interactions, small molecule chemical reactions, and interfaces from three-dimensional structures, pathways, and genetic interaction networks. BIND supports additional file formats to achieve compatibility with other database efforts, including the HUPO PSI Level 2. BIND's latest software spans over 2000 metadata fields and is constructed using the Java Enterprise Systems software platform. Protocols are provided for searching BIND via the Internet, as well as for viewing and exporting search results or individual records. Furthermore, a protocol is provided for visualizing biomolecular interactions within BIND or for transferring this information to the visualization tools Cytoscape and Cn3D. PMID- 18428772 TI - Using Chado to store genome annotation data. AB - Chado is a relational database schema that can be used to manage a wide variety of biological information, including genome annotation, genetic, phenotypic, and expression data. Its flexibility comes from its use of "ontologies," which are controlled vocabularies that describe data types and the relationships among them. By changing its ontologies, Chado can be customized to suit many different needs. Another aspect that gives Chado its flexibility is its use of a modular design, which means that users can choose to use only those features of Chado that are suitable for their needs. XORT is the main software tool used to move data in and out of Chado databases. XORT uses an XML-based file format for data import and export; this format is called ChadoXML, The protocols described in this chapter show how to use XORT and related software to import genome annotation data into Chado databases, and how to export data stored in Chado databases into different file formats for report and data mining purposes. PMID- 18428767 TI - Comparative protein structure modeling using Modeller. AB - Functional characterization of a protein sequence is one of the most frequent problems in biology. This task is usually facilitated by accurate three dimensional (3-D) structure of the studied protein. In the absence of an experimentally determined structure, comparative or homology modeling can sometimes provide a useful 3-D model for a protein that is related to at least one known protein structure. Comparative modeling predicts the 3-D structure of a given protein sequence (target) based primarily on its alignment to one or more proteins of known structure (templates). The prediction process consists of fold assignment, target-template alignment, model building, and model evaluation. This unit describes how to calculate comparative models using the program MODELLER and discusses all four steps of comparative modeling, frequently observed errors, and some applications. Modeling lactate dehydrogenase from Trichomonas vaginalis (TvLDH) is described as an example. The download and installation of the MODELLER software is also described. PMID- 18428773 TI - PubSearch and PubFetch: a simple management system for semiautomated retrieval and annotation of biological information from the literature. AB - For most systems in biology, a large body of literature exists that describes the complexity of the system based on experimental results. Manual review of this literature to extract targeted information into biological databases is difficult and time consuming. To address this problem, we developed PubSearch and PubFetch, which store literature, keyword, and gene information in a relational database, index the literature with keywords and gene names, and provide a Web user interface for annotating the genes from experimental data found in the associated literature. A set of protocols is provided in this unit for installing, populating, running, and using PubSearch and PubFetch. In addition, we provide support protocols for performing controlled vocabulary annotations. Intended users of PubSearch and PubFetch are database curators and biology researchers interested in tracking the literature and capturing information about genes of interest in a more effective way than with conventional spreadsheets and lab notebooks. PMID- 18428774 TI - Common file formats. AB - This appendix discusses a few of the file formats frequently encountered in bioinformatics. Specifically, it reviews the rules for generating FASTA files and provides guidance for interpreting NCBI descriptor lines, commonly found in FASTA files. In addition, it reviews the construction of GenBank, Phylip, MSF and Nexus files. PMID- 18428775 TI - Unix survival guide. AB - For a mixture of historical and practical reasons, much of the bioinformatics software discussed in this series runs on Linux, Mac OSX, Solaris, or one of the many other Unix variants. This appendix provides the novice with easy-to understand information needed to survive in the Unix environment. PMID- 18428776 TI - X Window System survival guide. AB - Logging in to a Unix system from a console typically initiates a graphical desktop environment that is similar to the Microsoft Windows and Apple Macintosh desktops. Logging in remotely to a Unix system, however, typically limits the user to a small text-only window, which is unable to launch graphical applications. This appendix describes the two main options for overcoming this obstacle: a Virtual Network Computer (VNC) and the X Window System. PMID- 18428777 TI - Glossary of bioinformatics terms. AB - This collection of terms and definitions commonly encountered in the bioinformatics literature will be updated periodically as Current Protocols in Bioinformatics grows. In addition, an extensive glossary of genetic terms can be found on the Web site of the National Human Genome Research Institute (http://www.genome.gov/glossary.cfm). The entries in that online glossary provide a brief written definition of the term; the user can also listen to an informative explanation of the term using RealAudio or the Windows Media Player. PMID- 18428778 TI - An introduction to hidden Markov models. AB - This unit introduces the concept of hidden Markov models in computational biology. It describes them using simple biological examples, requiring as little mathematical knowledge as possible. The unit also presents a brief history of hidden Markov models and an overview of their current applications before concluding with a discussion of their limitations. PMID- 18428779 TI - Using the Ensembl genome server to browse genomic sequence data. AB - The Ensembl genome Web browser (http://www.ensembl.org) provides a comprehensive source of automatic annotation of the human genome sequence (as well as other species of biomedical interest), with confirmed gene predictions that have been integrated with external data sources. This unit describes how to use the Ensembl browser, how to find your gene or protein of interest and get information and external links about them, and how to use the comparative genomic data. PMID- 18428780 TI - The UCSC Genome Browser. AB - The University of California Santa Cruz (UCSC) Genome Browser (genome.ucsc.edu) is a popular Web-based tool for quickly displaying a requested portion of a genome at any scale, accompanied by a series of aligned annotation "tracks". The annotations-generated by the UCSC Genome Bioinformatics Group and external collaborators-display gene predictions, mRNA and expressed sequence tag alignments, simple nucleotide polymorphisms, expression and regulatory data, and pairwise and multiple-species comparative genomics data. All information relevant to a region is presented in one window, facilitating biological analysis and interpretation. The database tables underlying the Genome Browser tracks can be viewed, downloaded, and manipulated using another Web-based application, the UCSC Table Browser. Users can upload personal data as custom annotation tracks in both browsers for research or educational use. This unit describes how to use the Genome Browser and Table Browser for genome analysis, download the underlying database tables, and create and display custom annotation tracks. PMID- 18428781 TI - Using the NCBI Map Viewer to browse genomic sequence data. AB - This unit includes an introduction to the Map Viewer, which describes how to perform a simple text-based search of genome annotations to view the genomic context of a gene, navigate along a chromosome, zoom in and out, and change the displayed maps to hide and show information. It also describes some of NCBI's sequence-analysis tools, which are provided as links from the Map Viewer. The Alternate Protocols describe different ways to query the genome sequence, and also illustrate additional features of the Map Viewer. Alternate Protocol 1 shows how to perform and interpret the results of a BLAST search against the human genome. Alternate Protocol 2 demonstrates how to retrieve a list of all genes between two STS markers. Finally, Alternate Protocol 3 shows how to find all annotated members of a gene family. PMID- 18428782 TI - Using galaxy to perform large-scale interactive data analyses. AB - While most experimental biologists know where to download genomic data, few have a concrete plan on how to analyze it. This situation can be corrected by: (1) providing unified portals serving genomic data and (2) building Web applications to allow flexible retrieval and on-the-fly analyses of the data. Powerful resources, such as the UCSC Genome Browser already address the first issue. The second issue, however, remains open. For example, how to find human protein coding exons with the highest density of single nucleotide polymorphisms (SNPs) and extract orthologous sequences from all sequenced mammals? Indeed, one can access all relevant data from the UCSC Genome Browser. But once the data is downloaded how would one deal with millions of SNPs and gigabytes of alignments? Galaxy (http://g2.bx.psu.edu) is designed specifically for that purpose. It amplifies the strengths of existing resources (such as UCSC Genome Browser) by allowing the user to access and, most importantly, analyze data within a single interface in an unprecedented number of ways. PMID- 18428783 TI - Assembling genomic DNA sequences with PHRAP. AB - The PHRAP assembly program provides rapid comparison, alignment, and assembly of large sets of DNA sequences. PHRAP compares sequences by searching for pairs of perfectly matching "words" or sequence regions that meet certain criteria. If a match is found, PHRAP then tries to extend the alignment into overlapping sections called contigs. PHRAP uses quality values produced by the PHRED basecaller to strike a balance between tolerance of discrepancies and prevention of stacking repeat sequences. The PHRAP assembly algorithm is generally used as part of the PHRED/PHRAP/Consed software suite for sequence analysis. This unit presents instructions for basic usage of the PHRAP assembler, including preparation of the input files (Support Protocols 1 and 2) and explanation of output files (Basic Protocols 1 and 2). Several command line options for changing the PHRAP assembly parameters are also discussed (Basic Protocol 3). PMID- 18428784 TI - Identifying structural noncoding RNAs using RNAz. AB - The functions of many noncoding RNAs and cis-acting regulatory elements of mRNAs depend on a defined RNA secondary structure. RNAz predicts such functional RNA structures on the basis of thermodynamic stability and evolutionary conservation of homologous sequences. It can be used to efficiently filter multiple alignments for noncoding RNA candidates in genomic screens. PMID- 18428785 TI - Validation of tandem mass spectrometry database search results using DTASelect. AB - DTASelect provides a means by which complex SEQUEST results can be filtered, organized, and viewed. A single sample may produce tens of thousands of tandem mass spectra. Manually perusing and selecting SEQUEST matches among such a mass of data carries a risk of inconsistency. DTASelect allows the user to set complex criteria for acceptance or rejection of individual spectrum results. It also features rules for dealing with multiple, identical peptide matches and for removing proteins that are insufficiently evidenced. It provides its sorted and filtered summary as HTML and text documents for easy review and also offers several auxiliary reports. DTASelect is a powerful tool for automatic analysis of complex mixture tandem mass spectrometry. PMID- 18428786 TI - Installation and use of the Computational Proteomics Analysis System (CPAS). AB - The community of proteomic researchers has been generating openly available software infrastructure for systematic proteomic data analyses and data management for the past several years. The Computational Proteomics Analysis System (CPAS) integrates many of these efforts into a single Web-based platform for mining liquid chromatography-tandem mass spectrometry (LC-MS/MS) proteomic experiments. CPAS incorporates several tools currently used in proteomic analysis, including the X! Tandem search engine and the PeptideProphet and ProteinProphet data mining tools. These tools and others are integrated into a single application built on the open-source LabKey platform, an extensible architecture for developing high-throughput biological applications. The CPAS analysis pipeline acts on data in standardized file formats, so that researchers may use CPAS with other search engines, including Mascot or SEQUEST, that follow a standardized procedure for reporting search engine results. All components of the CPAS toolkit are freely available at http://cpas.fhcrc.org. PMID- 18428787 TI - Using ProSight PTM and related tools for targeted protein identification and characterization with high mass accuracy tandem MS data. AB - ProSight PTM v2.0, neuroProSight, and the Sequence Gazer allow the identification and characterization of proteins from high mass accuracy tandem mass spectrometric data of intact proteins and large peptides. Input data consists of one or more neutral precursor ion masses and a set of neutral b/y or c/z(.) fragment ions masses. This data is compared against "shotgun annotated" proteome databases or known protein sequences. With these tools it is possible to not only identify unknown proteins, but to determine the location of post-translational modifications (PTM) with 100% sequence coverage. Collectively, the tools create a search environment that allows five different search modes, including absolute mass and sequence tag searching, which are conveniently employed via a graphical user interface. Data management and chemical noise reduction tools are also available. These tools provide a complete environment for the identification and characterization of proteins from high resolution tandem mass spectrometry of intact proteins and large peptides. PMID- 18428788 TI - Introduction to cheminformatics. AB - Cheminformatics is a relatively new field of information technology that focuses on the collection, storage, analysis, and manipulation of chemical data. The chemical data of interest typically includes information on small molecule formulas, structures, properties, spectra, and activities (biological or industrial). Cheminformatics originally emerged as a vehicle to help the drug discovery and development process, however cheminformatics now plays an increasingly important role in many areas of biology, chemistry, and biochemistry. The intent of this unit is to give readers some introduction into the field of cheminformatics and to show how cheminformatics not only shares many similarities with the field of bioinformatics, but that it can also enhance much of what is currently done in bioinformatics. PMID- 18428789 TI - In silico drug exploration and discovery using DrugBank. AB - DrugBank is a fully curated drug and drug target database that contains information on nearly 5000 drugs, including > 1200 FDA-approved small molecule and biotech drugs as well as >3200 experimental drugs. Additionally, more than 14,000 protein or drug target sequences are linked to these drug entries. DrugBank is primarily focused on providing both the query/search tools and the biophysical data needed to facilitate drug discovery and drug development. This unit provides readers with a detailed description of how to effectively use the DrugBank database and how to navigate through the DrugBank Web site. It also provides specific examples of how to find chemical homologs of potential drug leads and how to identify potential drug targets from newly sequenced pathogens. The intent of this unit is to give readers some introduction into the field of cheminformatics (the study of chemical information) and to show how cheminformatics can be seamlessly integrated into the field of bioinformatics. PMID- 18428790 TI - Using PhyloCon to identify conserved regulatory motifs. AB - Understanding gene regulation has been and remains one of the major challenges for the molecular biology community. Gene regulation is mediated by a variety of short DNA sequences called regulatory elements, which include transcription factor binding sites. A first step toward understanding gene regulation is the identification of regulatory elements present in the genome. This challenge has been defined as the "motif finding problem" in the field of computational biology. Over the past 20 years, many algorithms have been developed to tackle the motif finding problem computationally. The PhyloCon algorithm, developed in 2003, is one of the first motif finding algorithms that take advantage of two important data resources, i.e., phylogenetic conservation and gene co-regulation, to improve the efficiency of motif identification in biological datasets. This unit presents basic protocols to obtain, install, and apply the PhyloCon program and discusses the underlying algorithm and how to interpret the results. PMID- 18428791 TI - Using geneid to identify genes. AB - This unit describes the usage of geneid, an efficient gene-finding program that allows for the analysis of large genomic sequences, including whole mammalian chromosomes. These sequences can be partially annotated, and geneid can be used to refine this initial annotation. Training geneid is relatively easy, and parameter configurations exist for a number of eukaryotic species. Geneid produces output in a variety of standard formats. The results, thus, can be processed by a variety of software tools, including visualization programs. Geneid software is in the public domain, and it is undergoing constant development. It is easy to install and use. Exhaustive benchmark evaluations show that geneid compares favorably with other existing gene finding tools. PMID- 18428792 TI - Maximum-likelihood analysis using TREE-PUZZLE. AB - TREE-PUZZLE provides a means to analyze and reconstruct evolutionary relationships and trees based on quartets, i.e., groups of four sequences. Basic Protocol 1 explains how to reconstruct trees based on the maximum-likelihood principle and quartet puzzling. Basic Protocol 2 discusses likelihood mapping, a method to visualize phylogenetic content in a multiple sequence alignment. Basic Protocol 3 explains how to compare tree topologies using different tests. PMID- 18428793 TI - Analysis of expression data: an overview. AB - After providing a brief introduction to microarray chips and experimental details, this overview discusses analysis techniques. Data analysis from microarray experiments generally involves two parts: acquiring and normalizing the data, and interpreting it. This unit focuses mostly on the latter, as it is less technology-specific. PMID- 18428794 TI - Structure-based pKa calculations using continuum electrostatics methods. AB - Electrostatic free energy is useful for correlating structure with function in proteins in which ionizable groups play essential functional roles. To this end, the pK(a) values of ionizable groups must be known and their molecular determinants must be understood. Structure-based calculations of electrostatic energies and pK(a) values are necessary for this purpose. This unit describes protocols for pK(a) calculations with continuum electrostatics methods based on the numerical solution of the linearized Poisson-Boltzmann equation by the method of finite differences. Critical discussion of key parameters, approximations, and shortcomings of these methods is included. Two protocols are described for calculations with methods modified empirically to maximize agreement between measured and calculated pK(a) values. Applied judiciously, these methods can contribute useful and novel insight into properties of surface ionizable groups in proteins. PMID- 18428795 TI - Flexible ligand docking with Glide. AB - Glide is a ligand docking program for predicting protein-ligand binding modes and ranking ligands via high-throughput virtual screening. Glide utilizes two different scoring functions, SP and XP GlideScore, to rank-order compounds. Three modes of sampling ligand conformational and positional degrees of freedom are available to determine the optimal ligand orientation relative to a rigid protein receptor geometry. This unit presents protocols for flexible ligand docking with Glide, optionally including ligand constraints or ligand molecular similarities. PMID- 18428796 TI - Installing and configuring CMap. AB - CMap is a Web-based tool for viewing comparisons between biological maps of different types. It allows for correspondences to be displayed between maps as disparate as genetic and physical maps. The protocols in this unit comprise a guide to installing, configuring, and importing data into CMap. PMID- 18428797 TI - Using the Generic Genome Browser (GBrowse). AB - A genome browser is software that allows users to visualize DNA, protein, or other sequence features within the context of a reference sequence, such as a chromosome or contig. The Generic Genome Browser (GBrowse) is an open-source browser developed as part of the Generic Model Organism Database project (Stein et al., 2002). GBrowse can be configured to display genomic sequence features for any organism and is the browser used for the model organisms Drosophila melanogaster (Grumbling and Strelets, 2006) and Caenorhabditis elegans (Schwarz et al., 2006), among others. The software package can be downloaded from the web and run on a Windows, Mac OS X, or Unix-type system. Version 1.64, as described in this protocol, was released in November 2005, but the software is under active development and new versions are released about every six months. PMID- 18428799 TI - Directional conductivity in SWNT-collagen-fibrin composite biomaterials through strain-induced matrix alignment. AB - Composite biomaterials incorporating fibroblast cells, collagen Type I, fibrin, and 2 wt % carboxylated SWNT were created, and their properties were compared with similar control constructs without SWNT. Alignment of the matrix was stimulated by application of 8% cyclic strain for three 12-h periods over three days. All constructs underwent cell-mediated gel compaction to 15-20% of their initial volume, which was not affected by SWNT loading. Mechanical strain increased the rate of compaction, and strained constructs were significantly more compacted than unstrained controls by day 3. Cell viability and morphology were similar in both control and SWNT-loaded constructs, but unstrained samples exhibited a more stellate appearance with more numerous cellular projections. Application of mechanical strain caused clear alignment of both the cells and matrix in the direction of the applied strain. Bioimpedance measurements showed that SWNT loading increased the electrical conductivity of composite constructs, and that mechanically-induced alignment of the matrix/SWNT caused a further increase in conductivity. These results demonstrate that SWNT can be used to augment the electrical properties of 3D protein hydrogels, and that anisotropy in the matrix further enhances these properties. Such electrically conductive biopolymers may have a variety of applications in tissue engineering and biosensor development. PMID- 18428798 TI - Evaluation of the physical stability of the EC5 domain of E-cadherin: effects of pH, temperature, ionic strength, and disulfide bonds. AB - The development of protein drugs has been hampered by difficulties in formulating them due to their inherent chemical and physical stability, which could generate problems during the late stages of development. Thus, a basic understanding of the effect of structural features on the physicochemical stability of proteins can provide fundamental solutions to the formation of proteins. In this work, the physical stability of the EC5 protein under variable pH, temperature, and ionic strength and the role of the disulfide bond on the physical stability of EC5 were evaluated. All spectroscopic measurements were integrated in empirical phase diagrams, and these diagrams showed the stable and unstable regions of EC5. The native EC5 is more stable at high than at low ionic strength in a wide pH range during temperature elevation to 70 degrees C. The empirical phase diagrams also show that the reduced EC5 has lower stability than the parent EC5. The reduced EC5 has secondary structure only at pH 3 and 4 and is unfolded at other pH values. Finally, the reduced EC5 rapidly forms a precipitate at pH 4 and 5 upon heating. In conclusion, this study shows that ionic strength and the presence of the disulfide bonds are critical for the stability of EC5. PMID- 18428800 TI - Fabrication and in vitro degradation of porous fumarate-based polymer/alumoxane nanocomposite scaffolds for bone tissue engineering. AB - In this work, the fabrication and in vitro degradation of porous fumarate based/alumoxane nanocomposites were evaluated for their potential as bone tissue engineering scaffolds. The biodegradable polymer poly (propylene fumarate)/propylene fumarate-diacrylate (PPF/PF-DA), a macrocomposite composed of PPF/PF-DA and boehmite microparticles, and a nanocomposite composed of PPF/PF-DA and surface-modified alumoxane nanoparticles were used to fabricate porous scaffolds by photo-crosslinking and salt-leaching. Scaffolds then underwent 12 weeks of in vitro degradation in phosphate buffered saline at 37 degrees C. The presence of boehmite microparticles and alumoxane nanoparticles in the polymer inhibited scaffold shrinkage during crosslinking. Furthermore, the incorporation of alumoxane nanoparticles into the polymer limited salt-leaching, perhaps due to tighter crosslinking within the nanocomposite. Analysis of crosslinking revealed that the acrylate and overall double bond conversions in the nanocomposite were higher than in the PPF/PF-DA polymer alone, though these differences were not significant. During 12 weeks of in vitro degradation, the nanocomposite lost 5.3% +/- 2.4% of its mass but maintained its compressive mechanical properties and porous architecture. The addition of alumoxane nanoparticles into the fumarate based polymer did not significantly affect the degradation of the nanocomposite compared with the other materials in terms of mass loss, compressive properties, and porous structure. These results demonstrate the feasibility of fabricating degradable nanocomposite scaffolds for bone tissue engineering by photo crosslinking and salt-leaching mixtures of fumarate-based polymers, alumoxane nanoparticles, and salt microparticles. PMID- 18428801 TI - Synthesis of a 4-selenothymidine phosphoramidite and incorporation into oligonucleotides. AB - The detailed synthetic protocol for a 4-selenothymidine phosphoramidite and its use to prepare modified oligonucleotides is described here. The Se phosphoramidite synthesis was achieved by developing a useful protection and deprotection system for the selenium functionality. The coupling reaction of the Se-phosphoramidite during solid-phase oligonucleotide synthesis is quantitative, and the oligonucleotides containing the Se-modification are stable. Based on crystal structure analysis, the selenium-modified oligonucleotides retain base pairing like their native counterparts, and the derivatized DNA structure is virtually identical to the native structure. This achievement will present a novel opportunity for structural studies of nucleic acids and their protein complexes, because selenium can resolve the phase problem in macromolecular X-ray crystallography. In addition, this atom-specific replacement of oxygen with selenium will provide a useful tool for investigating biochemical and biophysical properties of nucleic acids and their protein complexes. PMID- 18428802 TI - Detection of aberrant 2'-5' linkages in RNA by anion exchange. AB - Formation of aberrant 2'-5' linkages can unintentionally occur in chemical synthesis of RNA. These linkages may arise by phosphoryl migration during deprotection, release, or subsequent steps during manufacture of therapeutic RNA. Their presence has been linked to a number of biochemical activities, so their potential for contribution to "off-target" effects is significant. Assaying for their presence, to ensure safe and effective therapeutic activity, is not straightforward. Since these linkages do not alter the RNA mass or the ionic or hydrophobic character of the product, confirmation of their presence or absence is not readily addressable by conventional chromatographic, electrophoretic, or mass spectrometric techniques. Since 2'-5' linkages are known to alter RNA solution conformation, they may also alter stationary phase interactions. A method for identifying the presence of these aberrant linkages by pellicular anion-exchange HPLC is presented in this unit. PMID- 18428803 TI - Chromophoric 5'-O-Silyl protection of N-protected 2'-ACE ribonucleosides for solid-phase RNA synthesis. AB - Recent advances in the understanding of the pivotal roles played by endogenous small RNAs in gene regulation have resulted in a substantial and rapidly growing market for synthetic RNA. 5'-Silyl-2'-ACE chemistry has proven to be a robust and reliable technology for the synthesis of oligoribonucleotides. This unit describes an important improvement to this chemistry, by adding a cycle-to-cycle traceability analogous to that inherent in 5'-dimethoxytrityl-based approaches. This is achieved by first regioselectively introducing a 5'-alkynylsilyl protecting group onto the 2'-ACE-protected nucleosides. The 5'-alkynylsilyl group is then reacted with an azide derivative of the chromophore Disperse Red 1, which enables spectrophotometric interrogation of each coupling step following 5' deprotection. Finally, the protected nucleosides are elaborated into their 3' phosphoramidite derivatives for use in solid-phase RNA synthesis. PMID- 18428804 TI - Synthesis of dimeric 2-amino-1,8-naphthyridine and related DNA-binding molecules. AB - The synthetic protocols for dimeric 2-amino-1,8-naphthyridine and related compounds are described in this unit. These compounds represent a novel class of compounds that bind selectively to mismatched base pairs. The compounds consist of two main components: the heterocycles and a linker. Connecting two heterocycles by a linker was found to modulate the binding selectivity. This unit describes the synthesis of dimeric 2-amino-1,8-naphthyridine (which binds to the G-G mismatch), naphthyridine-azaquinolone (for the G-A mismatch), N-alkyl naphthyridine dimer (for the C-C mismatch), and naphthyridine carbamate dimer (for G-G mismatches in the (CGG)(n) trinucleotide repeat). Protocols for connecting the short linker to these molecules providing the compounds suitable for immobilization on solid surface are also described. PMID- 18428805 TI - Theoretical principles of in vitro selection using combinatorial nucleic acid libraries. AB - A new paradigm for drug discovery and biological research has developed from technologies that integrate combinatorial chemistry with rounds of selection and amplification, a technique called in vitro selection or systematic evolution of ligands by exponential enrichment (SELEX). This overview unit discusses nucleic acid libraries that can be used, affinity probability distributions, an equilibrium model for SELEX, and optimal conditions including concentrations and signal-to-noise ratios. PMID- 18428806 TI - Characteristics of nucleic acids. AB - This appendix is a compilation of information from Current Protocols in Molecular Biology (John Wiley & Sons, Inc.) and provides information about the major nucleoside/nucleotide constituents and structures for A-, B-, and Z-DNA. Tables and figures provide (1) experimentally useful properties of the nucleoside/nucleotide building blocks; (2) chemical structures of the nucleosides; (3) aspects of nucleotide stereochemistry that are important to an understanding of base pairing and secondary structure; (4) alternative (non Watson-Crick) base pairings; and (5) the three best characterized helix structures for DNA: A-, B-, and Z-DNA. PMID- 18428807 TI - IUPAC-IUB joint commission on biochemical nomenclature abbreviations and symbols for the description of conformations of polynucleotide chains. AB - This appendix presents nomenclature rules that are as close as possible to the originally published version and are reproduced here with permission from the European Journal of Biochemistry. A web-based version prepared by G.P. Moss is also available. PMID- 18428808 TI - Nucleoside and nucleotide nomenclature. AB - Current nomenclature in the area of nucleosides, nucleotides, and nucleic acids comprises a mixture of (1) common names that have gained official recognition, (2) guidelines that have been derived and officially recommended by the International Union of Pure and Applied Chemistry (IUPAC)/International Union of Biochemistry and Molecular Biology (IUBMB), and (3) evolving usage that is derived by individual scientists and laboratories and subjected to peer review through publication. A working group was commissioned in 1998 by IUBMB to review guidelines for nucleotide (including oligonucleotide) nomenclature. As those guidelines are developed and made available, they will be referenced in future updates of this appendix. The main purpose of this appendix is to provide pertinent references that will direct the reader to the relevant guidelines or evolving nomenclature as described in the literature. When additional suggestions or guidance are appropriate, those comments are included as well. PMID- 18428809 TI - Common buffers and stock solutions. AB - This appendix describes the preparation of buffers and reagents used in the manipulation of nucleic acids. PMID- 18428810 TI - References to commonly used techniques. AB - Many of the protocols in this manual assume a knowledge of basic (chemical and biochemical) techniques. Although some of these are outside the scope of Current Protocols in Nucleic Acid Chemistry (CPNC), several basic methods can be found in this volume, either as support protocols or as part of longer protocols. While tailored to the particular goals of the units in which they appear, such protocols can be adapted by the trained researcher to suit the needs of a particular laboratory. If additional explanation or details for molecular techniques are required, the reader is advised to consult Current Protocols in Molecular Biology (CPMB). To facilitate this cross-referencing, relevant CPMB units are cited throughout CPNC. This appendix lists some commonly used techniques described in CPNC and indicates where they can be found in CPNC. PMID- 18428811 TI - Denaturing polyacrylamide gel electrophoresis. AB - Thin polyacrylamide gels that contain a high concentration of urea as a denaturant are capable of resolving short (<500 nucleotides) single-stranded fragments of DNA or RNA that differ in length by as little as one nucleotide. Such gels are uniquely suited for nucleic acid sequence analysis, which is required, for instance, for all footprinting protocols. Thicker gels are often used to purify oligonucleotides. This appendix describes the pouring, running, and processing of a typical sequencing gel, which is 40 cm long with a uniform thickness of 0.4 mm, containing 7 M urea and 4% to 8% acrylamide. PMID- 18428812 TI - Introduction to the synthesis and purification of oligonucleotides. AB - Modern nucleic acid synthesizers utilize phosphite triester chemistries that employ stable phosphoramidite monomers to build a growing polymer. These robust reactions allow easy generation of specific oligodeoxyribo- and oligoribonucleotides with a variety of labels, modified linkages, and nonstandard bases. Strategies are given for the maximization of synthetic yield, the generation of sequences containing site-specific modifications, and the isolation of synthetic oligonucleotides. Protocols describe monitoring the progress of synthesis via the trityl assay and methods for deprotection. PMID- 18428813 TI - Thin-layer chromatography. AB - TLC is used extensively in nucleic acid chemistry to monitor the progress of chemical reactions, to assay fractions collected from a larger chromatographic separation (e.g., column chromatography), and to determine optimal conditions prior to column chromatography. This unit describes methods for spotting test compounds onto a TLC plate, developing the plate in a suitable solvent system, visualizing the results, and calculating the retention factor (Rf). Candidate compounds can be co-spotted for identification without relying on Rf values. PMID- 18428814 TI - Column chromatography. AB - Column chromatography is used extensively in nucleic acid chemistry to purify or characterize products from chemical reactions. This appendix focuses on separations using silica gel or alumina. The procedures described include loading the column, assembling the apparatus, separating compounds and collecting fractions, and optimizing chromatography conditions. PMID- 18428815 TI - Useful nucleic acid chemistry web sites. AB - This appendix provides a listing of selected internet resources found to be of particular value by the CPNC editorial board; it is not intended to be a complete listing of all web resources cited in this manual. In addition to the sites listed here, the Internet Resources sections at the end of selected units suggest specialized sites pertaining to the contents of those units. PMID- 18428816 TI - Palladium-mediated C5 substitution of pyrimidine nucleosides. AB - One of the most efficient ways to link a reporter group to oligonucleotides is through the incorporation of a modified nucleoside during automated oligonucleotide synthesis. To be useful, it is important that the reporter group not interfere in hybridization reactions. This unit describes two linkers that can be used for the incorporation of a reporter group at the C5 position of deoxyuridine: a flexible aminoethylthioether linker, and a rigid amidopropynyl linker. The latter is sufficiently long and positioned so that the reporter group lies outside the major groove of the DNA duplex. PMID- 18428817 TI - Enzymatic synthesis of M(1)G-deoxyribose. AB - Adducts formed between electrophiles and nucleic acid bases are believed to play a key role in chemically induced mutations and cancer. M(1)G-dR is an endogenous exocyclic DNA adduct formed by the reaction of the dicarbonyl compound malondialdehyde with a dG residue in DNA. It is an intermediate in the synthesis of a class of modified oligodeoxyribonucleotides that are used to study the mutagenicity and repair of M(1)G. This unit presents methods for synthesizing M(1)G-dR by enzymatic coupling. PMID- 18428818 TI - Synthesis of N2-substituted deoxyguanosine nucleosides from 2-fluoro-6-O (trimethylsilylethyl)-2'-deoxyinosine. AB - Syntheses of N2-substituted nucleosides have been studied for many years, primarily with ribonucleosides. However, the primary route to these compounds requires acidic conditions that are too vigorous for the acid-labile deoxyribonucleosides. The current strategy takes advantage of methods for low temperature, nonaqueous diazotization of ribosides in organic solvents using t butyl nitrate as the diazotizing agent and HF/pyridine as the fluoride source for the preparation of a 2-fluoro-2-deoxyinosine derivative that can be used to synthesize N2-substituted deoxyguanosine. PMID- 18428819 TI - Unnatural nucleosides with unusual base pairing properties. AB - Synthetic modified nucleosides designed to pair in unusual ways with natural nucleobases have many potential applications in nucleic acid biochemistry. This overview lays the foundation for future protocol units on synthesis and application of unnatural bases, with particular emphasis on unnatural base analogs that mimic natural bases in size, shape, and biochemical processing. Topics covered including base pairs with alternative H-bonding schemes, hydrophobic base pairs, degenerate bases, universal nucleosides, and triplex constituents. PMID- 18428820 TI - Analysis of oligonucleotides by matrix-assisted laser desorption/ionization time of-flight mass spectrometry. AB - MALDI-MS is one of the most useful techniques available for determining biomolecule mass. It offers high mass accuracy, good sensitivity, simplicity, and speed. Because singly charged ions of oligonucleotides are typically observed, MALDI-MS spectra are easy to interpret. This unit presents protocols for sample preparation and purification, matrix preparation, and matrix/analyte sample preparation. It provides an introduction to the instrumentation and its calibration, and a discussion of some of the useful applications of MALDI-MS analysis in the study of oligonucleotides. This technique is typically used for 120-mer or smaller oligonucleotides. PMID- 18428821 TI - Analysis of oligonucleotides by electrospray ionization mass spectrometry. AB - Because of the high molecular weights and thermal lability of biomolecules such as nucleic acids and protein, they can be difficult to analyze by mass spectrometry. Such analyses require a "soft" ionization method that is capable of generating intact molecular ions. In addition, most mass analyzers have a limited upper mass range that is not sufficient for studying these large molecules. ESI MS can be used to analyze molecules with a molecular weight that is larger than the mass-to-charge ratio limit of the analyzer. This unit describes how ESI allows for analysis of high-molecular-weight compounds through the generation of multiply charged ions in the gas phase. It discusses analyzer configurations, solvent selection, and gives protocols for sample preparation. For applications of ESI-MS, the unit discusses molecular weight determination and gives protocols for sequencing and for analyzing oligonucleotide modifications. PMID- 18428822 TI - Overview of purification and analysis of synthetic nucleic acids. AB - Synthetic nucleic acids are produced routinely for a wide variety of applications, including biological and chemical research, and diagnostic or therapeutic applications. To ensure an adequate level of quality and purity, rapid and convenient analytical methods are necessary. This unit discusses basic principles to guide in the selection of appropriate purification and analysis protocols. PMID- 18428823 TI - Polyacrylamide gel electrophoresis (PAGE) of synthetic nucleic acids. AB - Protocols are given for analysis of oligonucleotides by PAGE, using either methylene blue staining or radiolabeling to mark the oligonucleotide. In addition, a separate protocol is provided for purification by PAGE. PMID- 18428824 TI - Analysis and purification of synthetic nucleic acids using HPLC. AB - HPLC is a powerful and popular method for analyzing and purifying biomolecules. Reversed-phase HPLC allows a high-capacity method for purification, and uses volatile buffer systems that simplify product recovery. Anion-exchange HPLC provides better resolution and a more predictable elution pattern. This unit presents protocols that are optimized for HPLC of oligonucleotides. Because of the resolution limits of both reversed-phase and anion-exchange HPLC, it can be used for oligonucleotides of up to approximately 50 nt in length. PMID- 18428825 TI - Base composition analysis of nucleosides using HPLC. AB - In this protocol, nuclease digestion of an oligonucleotide is followed by dephosphorylation and HPLC analysis of the monomers on a reversed-phase C18 column. This method can be used to detect and quantitate a wide variety of nucleobase modifications in oligonucleotides. Integrated areas of the nucleoside chromatogram give precise quantitation of nucleoside composition when the relative extinction coefficient cofactors are applied to the sum of the areas of the four bases. The protocol is also useful for analysis of oligonucleotides containing conjugated moieties and carbohydrate modifications. PMID- 18428826 TI - Cartridge methods for oligonucleotide purification. AB - Protocols are given for purification of oligonucleotides by dimethoxytrityl sensitive and affinity desalting methods. The protocols are applicable for many of the convenient disposable products available for rapid oligonucleotide purification, clean-up by selective adsorption, and elution on solid-phase media. Many of these products are prepackaged, single-use cartridges or columns filled with affinity or size-exclusion media. PMID- 18428827 TI - Analysis of oxidized DNA fragments by gel electrophoresis. AB - Polyacrylamide gel electrophoresis is used to define and quantify products of deoxyribose oxidation in DNA, based on the unique electrophoretic mobility of DNA fragments possessing deoxyribose oxidation products on their termini. This approach allows initial estimation of the chemistry. Once the chemical identity of damage products has been confirmed, this technique allows sensitive quantitation of the various damage products. PMID- 18428828 TI - RNA folding pathways. AB - A general overview of the questions and problems in RNA folding is presented. Topics include the differences in the folding problems/questions that apply to RNA versus proteins, methods for determination of final structures, folding versus unfolding, resolution of space and time, and conformational switching. PMID- 18428829 TI - Thermal methods for the analysis of RNA folding pathways. AB - Once a model of the secondary structure of an RNA has been deduced, thermal melting analysis can be used to determine whether the model accounts for all intramolecular interactions of the RNA, or whether noncanonical and tertiary interactions make the structure more stable than predicted, or link parts of the structure in unexpected ways. It is also useful to determine the pH, salt, and temperature ranges under which the RNA adopts a stably folded structure, or to analyze unfolding pathways. This unit discusses sample preparation, instrumentation, and theoretical background. It also provide a sample analysis of tRNA unfolding. PMID- 18428830 TI - Probing RNA folding pathways by RNA fingerprinting. AB - This unit provides protocols for using native polyacrylamide gel electrophoresis to distinguish folding and unfolding conformers of RNA. It is useful for studying conformers that can exchange in a period of minutes or seconds, and that are thus difficult to study by solution-based methods. Conformers that have been separated and immobilized in the gel matrix can be used to studying catalytic activity with or without being eluted from the gel. The method can be applied to a wide varied of catalytic RNAs and RNA-protein complexes. PMID- 18428831 TI - Characterization of tertiary folding of RNA by circular dichroism and urea. AB - CD spectroscopy can be used to monitor RNA tertiary folding transitions that may not be observable by absorbance spectroscopy. With the use of computer-controlled titrators, data can be acquired rapidly, and accurate thermodynamic properties can be obtained over a wide variety of conditions. Thus, CD spectroscopy provides a useful complement to site-resolved or chemical modification methods. PMID- 18428832 TI - Time-resolved hydroxyl radical footprinting of RNA with X-rays. AB - RNA tertiary structure and protein interactions can be detected by protection from hydroxyl radical cleavage. Generation of hydroxyl radicals with a synchrotron X-ray beam provides analysis on a short timescale (50 msec to 100 sec), which enables the structures of folding intermediates or other transient conformational states to be determined. This unit provides detailed instructions on the use of the synchrotron beamline for hydroxyl radical footprinting. PMID- 18428833 TI - Rapid magnesium chelation as a method to study real-time tertiary unfolding of RNA. AB - This unit describes a method to measure the unfolding of RNA tertiary structure on a millisecond time scale. A stopped-flow spectrophotometer is used to measure the rate of unfolding induced by the addition of EDTA to an RNA whose tertiary structure has been stabilized in the presence of magnesium ions. Using this methodology, rate constants for unfolding of tertiary or secondary structure can be obtained over a range of temperatures, and these values can be used to construct Arrhenius and Eyring plots, from which activation energy, Arrhenius pre exponential factor, and enthalpy and entropy of activation can be obtained. These data provide information about the energy of the transition state and the energy barriers between secondary and tertiary structure, which is necessary for predicting RNA tertiary structure from secondary structure. PMID- 18428834 TI - Nucleobase protection of deoxyribo- and ribonucleosides. AB - Protecting groups for the imide/lactam function of thymine/uracil and guanine, respectively, prevent irreversible nucleobase modifications that may occur in the presence of alkylating or condensing reagents that are commonly used in nucleoside protection and oligonucleotide synthesis. This unit reviews these protecting groups, and also identifies protecting groups for the exocyclic amino function of cytosine, adenine, and guanine. The unit also explores recent trends in nucleobase protection that permit reliable oligonucleotide synthesis and removal of N-protecting groups under very mild conditions. PMID- 18428835 TI - Protection of 2'-hydroxy functions of ribonucleosides. AB - The main purpose of this article is to discuss 2'-protection in the context of effective oligoribonucleotide synthesis. Emphasis is placed on the 2'-protecting groups of choice in the synthesis of oligo-and polyribonucleotides, and the requirements that a protective group must satisfy to become the 2'-hydroxyl protecting group of choice. Finally, the unit discusses the issue of 2'-O-acyl and 2'-O-silyl group migration to the 3'-hydroxy function of ribonucleosides during protection, along with the consequences of the conditions used for their removal on the stability of internucleotide linkages. PMID- 18428836 TI - Protection of 5'-hydroxy functions of nucleosides. AB - The 5-hydroxy group is the primary hydroxy group of nucleosides. It is mandatory to protect 5-hydroxyls in all methods of oligonucleotide synthesis that require nucleoside synthons. This unit discusses a wide variety of acid-labile and base labile protecting groups, as well as enzymatic methods for 5-protection and deprotection. PMID- 18428837 TI - A base-labile protecting group (fluorenylmethoxycarbonyl) for the 5'-hydroxy function of nucleosides. AB - Many popular synthesis strategies look for appropriate 2'-O-protection methods to use in conjunction with 5'-O-trityl chemistry. In contrast, this unit describes the use of FMOC as a 5'-protecting group in conjunction with a ketal-type 2'-O protecting group, 4-methoxytetrahydropyran-4-yl (MTHP). The synthesis of all four 2'-O-MTHP-5'-O-FMOC-protected ribonucleosides and 5'-O-FMOC-2'-deoxythymidine is described, as is the preparation of the N-protected, 2'-O-MTHP-protected starting nucleosides. PMID- 18428838 TI - 2'-Hydroxyl-protecting groups that are either photochemically labile or sensitive to fluoride ions. AB - Protected ribonucleotide monomers are more difficult to obtain than their 2' deoxy counterparts because of the need to protect the 2'-hydroxy function. This unit describes the stepwise preparation of suitably 2'-protected ribonucleosides using two protecting groups: 2-nitrobenzyloxymethyl (NBOM) and tert butyldimethylsilyl (TBDMS). In addition, details are given for protecting the 5' hydroxyl and the nucleobase, yielding nucleosides that are easily converted to phosphoramidite or H-phosphonate derivatives for automated oligoribonucleotide synthesis. PMID- 18428839 TI - Deoxyribo- and ribonucleoside H-phosphonates. AB - Most methods for preparing H-phosphonate monoesters suffer from variable yields and are often incompatible with common protecting groups used in oligonucleotide synthesis. This unit describes four procedures that consistently give high yields of the desired products. Taken together, they provide an arsenal of phosphonylation procedures that it compatible with most common protecting groups. PMID- 18428840 TI - Deoxyribonucleoside phosphoramidites. AB - The detailed preparation of deoxyribonucleoside phosphoramidites bearing a 4-[N methyl-N-(2,2,2-trifluoroacetyl)amino]butyl group for P(III) protection is presented. The use of this group circumvents nucleobase alkylation during oligonucleotide deprotection. Two syntheses of phosphoramidites starting from either a phosphordichloridite precursor or a bis-(N,N diisopropylamino)chlorophosphine intermediate are described for the phosphinylation of suitably protected deoxyribonucleosides. PMID- 18428841 TI - Regioselective 2'-silylation of purine ribonucleosides for phosphoramidite RNA synthesis. AB - This unit describes high-yield procedures for protection of purine ribonucleosides based on a reaction that allows concomitant highly regioselective 2'-silylation and 3'-phosphitylation to give, in one step, monomers that are ready for H-phosphonate synthesis. For preparation of phosphoramidites, the H phosphonate monoester is cleaved without silyl migration to give intermediates ready for phosphitylation by standard methods. PMID- 18428842 TI - Solid-phase supports for oligonucleotide synthesis. AB - This unit begins with a discussion of the advantages and disadvantages of oligonucleotide synthesis using solid supports. The physical and chemical properties of solid-phase supports are discussed in terms of their suitability for oligonucleotide synthesis. In addition, the unit outlines the properties of linkers used for transient or permanent attachment of properly protected nucleosides to the derivatized support, as well as strategies for coupling nucleosides to linkers and conditions for the release of synthetic oligonucleotides from specific supports. PMID- 18428843 TI - Attachment of nucleosides to solid-phase supports. AB - Specific step-by-step instructions are given for coupling nucleosides to LCAA-CPG supports (supports consisting of a long-chain alkylamine linked to controlled pore glass). Protocols are given for a succinic acid linker and a hydroquinone O,O'-diacetic acid linker. The former is the most widely used linker arm, and the starting materials are widely available. The latter offers greater compatibility with base-sensitive sequence modifications and great synthetic throughput because it can be cleaved under milder and faster conditions. Additional guidelines are given for selecting a linker arm and coupling protocol. Almost any application requiring synthetic oligonucletodies can be satisfied using one of these linker arms. PMID- 18428844 TI - Synthetic strategies and parameters involved in the synthesis of oligodeoxyribonucleotides according to the phosphoramidite method. AB - The phosphoramidite approach has had a major impact on the synthesis of oligonucleotides. This unit describes parameters that affect the performance of this method for preparing oligodeoxyribonucleotides, as well as a number of compatible strategies. Milestones that led to the discovery of the approach are chronologically reported. Alternate strategies are also described to underscore the versatility by which these synthons can be obtained. Mechanisms of deoxyribonucleoside phosphoramidite activation, factors affecting condensation, and deprotection strategies are discussed. PMID- 18428845 TI - Strategies for oligoribonucleotide synthesis according to the phosphoramidite method. AB - Advances in oligoribonucleotide synthesis have lagged behind those in oligodeoxyribonucleotide synthesis because of the difficulty in identifying orthogonal protecting groups for the 2'- and 5'-hydroxyls. Adaptation of the phosphoramidite method for DNA synthesis to RNA synthesis has greatly improved our understanding of RNA. It allows site-specific introduction of modified nucleosides to any position in an RNA molecule, as well as introduction of variations at multiple sites in the molecule. This overview discusses issues that are relevant to RNA synthesis by the phosphoramidite approach, including supports used, activation of the ribonucleoside phosphoramidites, and protection of the nucleobase, phosphate, and 2'- and 5'-hydroxyls. PMID- 18428846 TI - Oligoribonucleotides with 2'-O-(tert-butyldimethylsilyl) groups. AB - The chemical synthesis of oligoribonucleotides on solid support is routinely performed via the phosphoramidite method. However, the additional 2-OH function of the ribofuranosyl sugar requires suitable protection during oligoribonucleotide synthesis. This unit describes methods for 2-OH protection using the TBDMS group. PMID- 18428847 TI - Synthesis of oligoribonucleotides using the 2-nitrobenzyloxymethyl group for 2' hydroxyl protection. AB - RNA can be rapidly synthesized using protected ribonucleoside phosphoramidite monomers that carry 2-nitrobenzyloxymethyl groups on their 2-hydroxyls. These protecting groups are removed from the oligoribonucleotide products by exposure to long-wave UV light. PMID- 18428848 TI - A brief history, status, and perspective of modified oligonucleotides for chemotherapeutic applications. AB - The advent of rapid and efficient methods of oligonucleotide synthesis has allowed the design of modified oligonucleotides that are complementary to specific nucleotide sequences in mRNA targets. Such modified oligonucleotides can be used to disrupt the flow of genetic information from transcribed mRNAs to proteins. This antisense strategy has been used to develop therapeutic oligonucleotides against cancer and various infectious diseases in humans. This overview reports recent advances in the application of oligonucleotides as drug candidates, describes the relationship between oligonucleotide modifications and their therapeutic profiles, and provides general guidelines for enhancing oligonucleotide drug properties. PMID- 18428849 TI - Modification of the 5' terminus of oligonucleotides for attachment of reporter and conjugate groups. AB - Reporter and conjugate groups can be added directly to the 5' terminus of oligonucleotides by appropriate modification. Conjugate groups can be used to increase the affinity of complementary strands, induce irreversible modification of target sequences, or enable sequences to recognize and permeate target cell membranes. This overview discusses the 5' modifications that can be used and strategies for the covalent attachment of ligands to the modified oligonucleotides. Step-by-step protocols for attachment of conjugate groups are given elsewhere in the series. PMID- 18428850 TI - Direct attachment of conjugate groups to the 5' terminus of oligodeoxyribonucleotides. AB - This unit gives protocols for the attachment of intercalating and photoreactive conjugate groups to oligodeoxyribonucleotides. Protocols are given for acridine- and psoralen-conjugated oligonucleotides, and include attachment of the linker, preparation of the phosphoramidite, coupling to the oligonucleotide, deprotection, purification, and characterization. PMID- 18428851 TI - Synthesis and characterization of chimeric 2-5A-DNA oligonucleotides. AB - This unit provides protocols for the synthesis and characterization of 2-5A antisense nucleic acids. These chimeric oligonucleotides consist of 2',5' phosphodiester-linked oligoadenylates ligated to 3',5'-deoxyribonucleotides and are readily prepared using phosphoramidite chemistry on CPG solid supports. The 3',5'-deoxyribonucleotide functions as the antisense domain to target a given mRNA sequence, while the 2',5'-phosphodiester-linked oligoadenylate serves to locally activate 2-5A-dependent RNase L, causing the targeted sequence to be cleaved. PMID- 18428852 TI - Attachment of reporter and conjugate groups to the 3' termini of oligonucleotides. AB - Conjugation of oligonucleotides at the 3 terminus is less common because this site is used for covalent linkage to solid-phase oligonucleotide synthesis supports. However, 3-oligonucleotide conjugates have several valuable physicochemical properties, including their ability to stabilize nucleic acid hybridization complexes and to retard the activity of exonucleases. This unit discusses methods for preparing oligonucleotides conjugated at the 3 terminus. PMID- 18428853 TI - 3'-modified oligonucleotides and their conjugates. AB - Solid-phase synthesis of 3'-aminoalkyl, 3'-thioalkyl, and 3'-polyethyleneglycol are described. The first two are important as specific points of attachment for a large variety of reporter groups; the latter is important because of its increased cell membrane permeability, which aids in the development of antisense drugs. These protocols cover details of the synthetic organic chemistry needed to make each solid support, as well as DNA synthesis, workup, characterization, and conjugation. PMID- 18428855 TI - Incorporation of halogenoalkyl, 2-pyridyldithioalkyl, or isothiocyanate linkers into ligands. AB - Ligands can be introduced at the 5' terminus of an oligonucleotide by adding a linker to the ligand and modifying the 5' terminus of the oligonucleotide. These are then reacted to give the ligand-oligonucleotide conjugate. The addition of appropriate linkers to ligands is described in this unit. 5'Modification of the oligonucleotide and the final reaction that produces the ligand-conjugated oligonucleotide are described elsewhere in the series. This approach is particularly useful when there is a limited amount of ligand available, when the ligand is sensitive to chemical conditions required for oligonucleotide deprotection, or when the ligand is weakly soluble in solvents required for phosphoramidite- or H-phosphonate-mediated oligonucleotide synthesis. PMID- 18428854 TI - Synthesis and purification of oligonucleotide N3'-->P5' phosphoramidates and their phosphodiester and phosphorothioate chimeras. AB - This unit describes the synthesis and purification of oligonucleotide N3'-->P5' phosphoramidates, wherein each 3'-oxygen is replaced by a 3'-amine in the 2' deoxyribose ring. The synthesis of required monomers and application of the method to preparation of phosphodiester- and phosphorothioate-containing chimera of phosphoramidate is also reported. PMID- 18428856 TI - Modification of the 5' terminus of oligodeoxyribonucleotides for conjugation with ligands. AB - Ligands can be introduced at the 5' terminus of an oligonucleotide by adding a linker to the ligand and modifying the 5' terminus of the oligonucleotide. These are then reacted to give the ligand-oligonucleotide conjugate. This unit describes the addition of carboxylated and aminoalkylated linkers, and phosphorothioate, phosphate, and masked thiol groups to the 5' terminus of an oligonucleotide. The addition of linkers to ligands and the final reaction that produces the ligand-conjugated oligonucleotide are described elsewhere in the series. This approach is particularly useful when there is a limited amount of ligand available, when the ligand is sensitive to chemical conditions required for oligonucleotide deprotection, or when the ligand is weakly soluble in solvents required for phosphoramidite- or H-phosphonate-mediated oligonucleotide synthesis. PMID- 18428857 TI - Engineering disulfide cross-links in RNA using thiol-disulfide interchange chemistry. AB - Protocols for postsynthetic modification of 2-amino-containing oligoribonucleotides with either an alkyl-phenyl disulfide or an alkyl thiol group are described. These groups react under mild conditions to form disulfide cross-links by thiol-disulfide interchange. These reactants do not form a disulfide bond when incorporated on opposite faces of a short continuous RNA helix, but do form disulfide bonds rapidly when they are placed in proximity. In addition, by incorporating these groups at various positions on large RNAs by semisynthesis, the dynamics of thermal motions can be detected. Such motions are believed to be linked to biological function, and the protocols presented in this unit are among the few simple ways to assess such dynamics. PMID- 18428858 TI - Chemical and enzymatic methods for preparing circular single-stranded DNAs. AB - Small circular oligonucleotides can be used for diagnostic, therapeutic, and laboratory purposes. These systems have gained considerable attention in recent years because they form unusually strong and specific complexes with RNA and DNA strands. Synthetic circular DNAs of 20 to 200 nucleotides can also serve as catalysts for amplified DNA and RNA synthesis by a rolling circle mechanism. This unit presents methods for synthesizing small circular oligonucleotides. These simple "one-pot" procedures are carried out using short DNA splints that hold the circle together until it is chemically or enzymatically ligated. PMID- 18428859 TI - Engineering specific cross-links in nucleic acids using glycol linkers. AB - One of the most convenient methods for generating oligonucleotides possessing intra- or interstrand cross-links is through incorporation of oligoethylene glycol bridges by solid-phase synthesis. The reagents are commercially available or can be synthesized in a few easy synthetic steps. Unlike many other DNA and RNA cross-links, aspects of the structural and thermodynamic impact of modifying nucleic acids with oligoethylene glycols have been studied. This unit covers protection, phosphitylation, and preparation of the glycol linker for oligonucleotide synthesis. PMID- 18428860 TI - Engineering disulfide cross-links in RNA via air oxidation. AB - This unit presents protocols for the synthesis of alkylthiol-modified ribonucleosides, their incorporation into synthetic RNA, and the formation of intramolecular disulfide bonds in RNA by air oxidation. The disulfide bonds can be formed in quantitative yields between thiols positioned in close proximity by virtue of either the secondary or tertiary structure of the RNA. Disulfide cross links are useful tools to probe solution structures of RNA, to monitor dynamic motion, to stabilize folded RNAs, and to study the process of tertiary structure folding. PMID- 18428861 TI - Use of electrophilic substitution to form site-specific cross-links in DNA. AB - This unit describes the synthesis, purification, and analysis of modified oligodeoxynucleotides containing 4-thio-2'-deoxyuridine, and the use of these modified oligodeoxynucleotides in conjunction with bis-electrophilic tethers to form site-specific cross-links with a complementary DNA target. These modified oligodeoxynucleotides provide a flexible tool for studies directed at biophysical investigation of oligonucleotide structure, as well as investigation of DNA hybridization, stability, and sequence-dependent base nucleophilicity. PMID- 18428862 TI - Probing RNA structure with chemical reagents and enzymes. AB - This unit provides thorough coverage of the most useful chemical and enzyme probes that can be used to examine RNA secondary and tertiary structure. Footprinting methods are presented using dimethyl sulfate, diethyl pyrocarbonate, ethylnitrosourea, kethoxal, CMCT, and nucleases. For chemical probes, both strand scission and primer extension detection protocols are included. PMID- 18428863 TI - Probing nucleic acid structure with shape-selective rhodium and ruthenium complexes. AB - In this unit, transition metal complexes are used as photochemical probes for the structure of RNA and DNA. The transition metal ion provides a rigid substitutionally inert framework and an octahedral geometry for ligand coordination. The complexes can be constructed to define shapes, symmetries, and functionalities that complement those of the nucleic acid target. Complex formation is easily detected by light-induced nucleic acid cleavage. The modular construction of the complexes makes it possible to generate probes to examine a wide variety of structural characteristics of nucleic acids. PMID- 18428864 TI - Probing RNA structure by lead cleavage. AB - Lead cleavage causes a transesterification reaction that breaks the 5',3' phosphodiester backbone of RNA, leaving a 2',3'-cyclic phosphate and a 5' hydroxyl. Since the efficiency of the reaction at the 2'-hydroxyl is related to steric and chemical constraints on particular 2'-hydroxyls embedded in the RNA, this reaction can be used to examine the structure of individual nucleotides within an RNA molecule. It is a sensitive probe of tertiary RNA structure, provided that Pb(2+)-binding sites are created in the tertiary structure. PMID- 18428865 TI - Probing nucleic acid structure with nickel- and cobalt-based reagents. AB - The use of nickel and cobalt reagents is presented for characterizing the solvent exposure of guanine residues in DNA and RNA. These reagents promote guanine oxidation in the presence of a peracid such as monopersulfate, and the extent of reaction indicates the steric and electronic environment surrounding the N7 and aromatic face of this residue. Since oxidation does not itself perturb target structure or induce strand scission, it is coupled with fragmentation by treatment with piperidine (for smaller polynucleotides) or termination of primer extension (for larger polynucleotides). PMID- 18428866 TI - Probing RNA structures with hydroxyl radicals. AB - Fe(II)-EDTA can be used to conveniently generate hydroxyl radicals to promote cleavage of RNA at nucleotide resolution. Two procedures are described, involving the generation of free radicals from solvated molecular oxygen and from hydrogen peroxide added to the RNA solution. Unlike other footprinting reagents, hydroxyl radicals cleave the sugar-phosphate backbone at every residue and thus provide uniform cleavage in a given RNA secondary structure. Because some positions become protected by tertiary folding, this reagent is useful for monitoring the global folding of RNA at equilibrium. PMID- 18428867 TI - Chemical reagents for investigating the major groove of DNA. AB - Chemical modification provides an inexpensive and rapid method for characterizing the structure of DNA and its association with drugs and proteins. Numerous conformation-specific probes are available, but most investigations rely on only the most common and readily available of these. The major groove of DNA is typically characterized by reaction with dimethyl sulfate, diethyl pyrocarbonate, potassium permanganate, osmium tetroxide, and, quite recently, bromide with monoperoxysulfate. This commentary discusses the specificity of these reagents and their applications in protection, interference, and missing contact experiments. PMID- 18428868 TI - Biophysical analysis of nucleic acids. AB - This overview unit provides a thorough overview of biophysical methods used for structure analysis, including X-ray diffraction, nuclear magnetic resonance, optical spectroscopy, theoretical and computational methods, and single-molecule methods. Advantages and disadvantages of the methods are compared. PMID- 18428870 TI - NMR determination of oligonucleotide structure. AB - This unit provides an overview of the use of NMR to determine oligonucleotide structure. It covers basic NMR spectral properties, acquisition of interproton distance restraints and torsion angle restraints, structure refinement, assessment of the quality of the structure obtained. Software programs used in the process are also described. PMID- 18428869 TI - Molecular modeling of nucleic acid structure: setup and analysis. AB - The last in a set of units by these authors, this unit addresses some important remaining questions about molecular modeling of nucleic acids. It describes how to choose an appropriate molecular mechanics force field; how to set up and equilibrate the system for accurate simulation of a nucleic acid in an explicit solvent by molecular dynamics or Monte Carlo simulation; and how to analyze molecular dynamics trajectories. PMID- 18428871 TI - Optical methods. AB - Equilibrium optical melting curves are used in this unit to determine thermodynamic parameters of nucleic acid complex formation. This section contains all of the relevant equations and a discussion of which are most appropriate to a given situation. Additionally, procedures are given for making preliminary determinations of molar extinction coefficients and for determining the number of oligonucleotides in a complex. The section on extinction coefficients is particularly essential to anyone needing to know solution concentrations. PMID- 18428872 TI - Calorimetry of nucleic acids. AB - This unit describes the application of differential scanning and isothermal titration calorimetry (DSC and ITC) to the study of the thermodynamics of nucleic acid structure. DSC is used to study order-disorder transitions. A single DSC profile provides a wealth of thermodynamic and extrathermodynamic information: transition enthalpy, entropy, free energy, heat capacity, the state of the transition (two-state vs. multistate), and the size of the cooperative unit. ITC is used to study hybridization of nucleic acids at constant temperature. Results can be used to determine the stoichiometry of the association reaction, the enthalpy of association, the equilibrium association constant, and the free energy and entropy of association. A thorough discussion is presented of the details required to obtain meaningful results, as well as relevant methods for analyzing the data produced. PMID- 18428873 TI - Molecular modeling of nucleic acid structure. AB - This unit is the first in a series of four units covering the analysis of nucleic acid structure by molecular modeling. This unit provides an overview of computer simulation of nucleic acids. Topics include the static structure model, computational graphics and energy models, generation of an initial model, and characterization of the overall three-dimensional structure. PMID- 18428874 TI - Methods to crystallize RNA. AB - Preparation of suitably large and well-ordered single crystals is usually the rate-limiting step in the determination of the three-dimensional structure of RNAs and their complexes with proteins by X-ray crystallography. This unit discusses a variety of experimental considerations for obtaining crystals of RNAs and RNA-protein complexes. Topics include design of crystallizable constructs, screening, and optimization of crystallization conditions. PMID- 18428875 TI - Recent advances in RNA structure determination by NMR. AB - Despite recent advances in the solution of NMR structures of RNA and RNA-ligand complexes, the rate limiting step remains the gathering of a large number of NOE and torsion restraints. Additional sources of information for structure determination of larger RNA molecules have recently become available, and it is possible to supplement NOE and J-coupling data with the measurement of dipolar couplings and cross-correlated relaxation rates in high-resolution NMR spectroscopy. PMID- 18428876 TI - Molecular modeling of nucleic acid structure: energy and sampling. AB - An overview of computer simulation techniques as applied to nucleic acid systems is presented. This unit expands an accompanying overview unit (UNIT 7.5) by discussing methods used to treat the energy and sample representative configurations. Emphasis is placed on molecular mechanics and empirical force fields. PMID- 18428878 TI - Determination of binding mode: intercalation. AB - A small molecule can be assumed to bind to DNA by intercalation between base pairs if it causes lengthening and unwinding of the DNA helix and undergoes changes in its spectral properties, such as DNA-induced hypochromism and quenching of its UV absorbance. DNA lengthening and unwinding can be determined from the change in viscosity of a solution of linear or plasmid DNA, respectively. Intercalation of a ligand can also be seen as a reduction in the UV/visible absorbance of the intercalator, as well as a shift in the absorbance maximum. PMID- 18428877 TI - Molecular modeling of nucleic acid structure: electrostatics and solvation. AB - This unit presents an overview of computer simulation techniques as applied to nucleic acid systems, ranging from simple in vacuo molecular modeling techniques to more complete all-atom molecular dynamics treatments that include an explicit representation of the environment. The third in a series of four units, this unit focuses on critical issues in solvation and the treatment of electrostatics. PMID- 18428879 TI - Determination of binding thermodynamics. AB - This unit serves as a starting point for exploring the thermodynamic properties of interactions between small molecules and DNA. It covers the determination of simple, apparent association/dissociation constants. The concentration of DNA bound ligand and free ligand are determined and a binding constant is extracted from these data. Data gathering and curve fitting are discussed. PMID- 18428880 TI - Design, synthesis, and amplification of DNA pools for in vitro selection. AB - Preparation of a random-sequence DNA pool is presented. The degree of randomization and the length of the random sequence are discussed, as is synthesis of the pool using a DNA synthesizer. Purification of a single-stranded pool and conversion to a double-stranded pool are presented as step-by-step protocols. Support protocols describe determination of the complexity and skewing of the pool, and optimization of amplification conditions. PMID- 18428881 TI - In vitro selection of RNA aptamers to a protein target by filter immobilization. AB - This unit describes the selection of aptamers from a pool of single-stranded RNA by binding to a protein target. Aptamers generated from this selection experiment can potentially function as protein inhibitors, and may find applications as therapeutic or diagnostic reagents. A pool of dsDNA is used to generate a ssRNA pool, which is mixed with the protein target. Bound complexes are separated from unbound reagents by filtration, and the RNA:protein complexes are amplified by a combination of reverse transcription, PCR, and in vitro transcription. PMID- 18428882 TI - Selection for catalytic function with nucleic acids. AB - For in vitro selection of catalytic polynucleotides, each new protocol must be designed to harness the desired catalytic activity to help propel the selection process itself. This unit gives guidelines for design of in vitro selection experiments for catalytic function. It outlines several representative protocols as examples of successful selection experiments, providing a conceptual basis for the design and implementation of new selective-amplification protocols for nucleic acids. PMID- 18428883 TI - Development of a universal nucleobase and modified nucleobases for expanding the genetic code. AB - This unit presents protocols for the synthesis and characterization of nucleosides with unnatural bases in order to develop bases for the expansion of the genetic alphabet or for nonselective pairing opposite natural bases. Protocols describe the design, synthesis, and characterization of unnatural base pairs involving 1-beta-D-2-deoxyribosyl-N- and -C-nucleosides. Determination of the thermodynamic and kinetic parameters of unnatural nucleosides is accomplished by incorporation into oligonucleotides and subsequent evaluation as described herein. PMID- 18428884 TI - Syntheses of specifically 15N-labeled adenosine and guanosine. AB - This unit describes the specific incorporation of (15)N into the N7 and amino positions of adenosine, and conversion of the adenosine to guanosine labeled at the N1, N7, and amino positions. Two variations of the procedures are also presented that include either (12)C or (13)C at the C8 position of adenosine, and (13)C at either the C8 or C2 position of guanosine. These (13)C tags permit the incorporation of two (15)N-labeled nucleosides into an RNA strand while ensuring that their nuclear magnetic resonance (NMR) signals can be distinguished from each other by the presence or absence of C-N coupling. While the major application of these specifically (15)N-labeled nucleosides is NMR, the additional mass makes them useful in mass spectrometry (MS) as well. The procedures can also be adapted to synthesize the labeled deoxynucleosides. A support protocol describes the synthesis of 7-methylguanosine. PMID- 18428885 TI - Synthesis of protected 2'-deoxy-2'-fluoro-beta-D-arabinonucleosides. AB - This unit describes in detail the preparation of protected 2'-deoxy-2' fluoroarabinonucleosides. These building blocks are required for the synthesis of 2'-deoxy-2'-fluoroarabinonucleic acid (2'F-ANA), an oligonucleotide analog exhibiting very promising antisense properties. The preparation of phosphoramidites from these building blocks and the synthesis of 2'F-ANA are described elsewhere in the manual. PMID- 18428886 TI - Use of fluorescence spectroscopy to elucidate RNA folding pathways. AB - This overview unit discusses fluorescence spectroscopy as a tool for studying RNA folding. Ribozymes and oligonucleotides can be labeled with a fluorescent probe and analyzed to give information about both slow and fast kinetic processes with real-time data acquisition. The unit discusses the advantages and disadvantages of various pendant probes and nucleotide analogs, the analytical methods that can be used, instrument setup, control experiments, and a variety of kinetic experiments that can be performed, such as determination of rate constants. PMID- 18428887 TI - Use of chemical modification to elucidate RNA folding pathways. AB - As discussed in this overview, chemical modification is sensitive to the accessibility of a nucleotide to the solvent, and many nucleotides become less accessible as an RNA folds into its structured form. Chemical modification reagents are therefore suitable for following RNA folding, and can be used to study the kinetics of structure formation on time scales ranging from minutes to hours. PMID- 18428888 TI - Key experimental approaches in DNA nanotechnology. AB - DNA nanotechnology combines unusual DNA motifs with sticky-ended cohesion to build polyhedral objects, topological targets, nanomechanical devices, and both crystalline and aperiodic arrays. The goal of DNA nanotechnology is control of the structure of macroscopic matter on the finest possible scale. Applications are expected to arise in the areas of X-ray crystallography, nanoelectronics, nanorobotics, and DNA-based computation. DNA and its close molecular relatives appear extremely well suited for these goals. This overview covers the generation of new DNA motifs, construction methods (synthesis, hybridization, phosphorylation, ligation), and a variety of methods for characterization of motifs, devices, and arrays. Finally, the use of DNA nanotechnology as a tool in biochemistry is discussed. PMID- 18428889 TI - Preparation of gold nanoparticle-DNA conjugates. AB - This unit describes the preparation of conjugates between nanometer-scale gold particles and synthetic oligonucleotides. Oligonucleotide-functionalized gold nanoparticles are finding increased use in both the construction of complex, tailored nanostructures and the optimization of DNA sequence analysis. The protocols in this unit outline the synthesis, purification, and characterization of nanoparticle-DNA conjugates for applications in nanotechnology and biotechnology. Separate procedures are presented for nanoparticles functionalized with just one or a few oligonucleotide strands and for nanoparticles functionalized with a dense layer of oligonucleotide strands. The different physical and chemical properties of these two types of conjugates are discussed, as are their stability and utility in different environments. PMID- 18428890 TI - Preparation of 2'-O-[(Triisopropylsilyl)oxy]methyl-protected ribonucleosides. AB - The [(triisopropylsilyl)oxy]methyl (TOM) group is a useful protecting group for the 2'-OH of ribonucleosides to be used for oligoribonucleotide synthesis by the phosphoramidite method. It is completely stable to all reaction conditions applied during assembly and the first deprotection step. It does not interfere with the coupling reaction and leads to very good coupling yields under DNA coupling conditions. The final cleavage occurs quantitatively without concomitant destruction of the RNA product. This unit describes the synthesis and characterization of 2'-O-TOM-5'-O-dimethoxytrityl-N-acetyl ribonucleosides in full detail. The TOM-group is introduced via a dibutyltin dichloride-mediated reaction into N-acetylated, 5'-O-dimethoxytritylated ribonucleosides. Support protocols describe the synthesis of N-acetylated, 5'-O-dimethoxytritylated adenosine and guanosine, as well as synthesis of the starting reagent [(triisopropylsilyl)oxy]methyl chloride (TOM-Cl). Preparation of the phosphoramidites and their use in solid-phase oligonucleotide synthesis are described elsewhere in the series. PMID- 18428891 TI - Chemical synthesis of RNA sequences with 2'-O-[(triisopropylsilyl)oxy]methyl protected ribonucleoside phosphoramidites. AB - The [(triisopropylsilyl)oxy]methyl (TOM) group is a useful protecting group for the 2'-OH of ribonucleosides to be used for oligoribonucleotide synthesis by the phosphoramidite method. It is completely stable to all reaction conditions applied during assembly and the first deprotection step. It does not interfere with the coupling reaction and leads to very good coupling yields under DNA coupling conditions. The final cleavage occurs quantitatively without concomitant destruction of the RNA product. This unit describes the synthesis of TOM phosphoramidites from 2'-O-TOM-5'-O-dimethoxytrityl-N-acetyl ribonucleosides, oligoribonucleotide assembly on an automated DNA synthesizer, and subsequent deprotection. Preparation of the TOM-protected ribonucleosides is presented elsewhere in the series. PMID- 18428892 TI - Conjugation of 5'-functionalized oligodeoxyribonucleotides with properly functionalized ligands. AB - This unit reports the synthesis of oligodeoxyribonucleotides covalently linked via their 5' termini to various ligands such as intercalating agents, reactive groups, or labels. Methods for incorporation of halogenoalkyl, isothiocyanate, and 2-pyridyldisulfide linkers onto ligands, and addition of amino, carboxyl, thiophosphate, phosphate, and masked thiol groups at the 5 terminus of an oligodeoxyribonucleotide are described elsewhere in the series. This unit reports procedures for coupling the ligands and oligonucleotides, as well as details for purification and characterization of the final products. PMID- 18428893 TI - Synthesis and purification of peptide nucleic acids. AB - Peptide nucleic acids (PNAs) are DNA analogs in which the normal phosphodiester backbone is replaced by 2-aminoethyl glycine linkages. Hybridization of PNAs with RNA or DNA follows normal rules for Watson-Crick base pairing and occurs with high affinity. Thus, PNAs are a promising choice for applications that benefit from high-affinity hybridization. They are assembled using techniques adapted from peptide chemistry. Protocols are given for both automated and manual synthesis of PNAs as well as their purification. The advantages of each method are discussed, as are the different monomers and reagents that are required. Additionally, protocols are given for adding peptides to PNAs (which can enhance hybridization or cell uptake of the PNA) and for adding a biotin label. PMID- 18428894 TI - Locked nucleic acids: synthesis and characterization of LNA-T diol. AB - Locked nucleic acids (LNAs) are RNA derivatives that have an O-methylene linkage between the 2 and 4 positions of the ribose. This leads to exceptionally high affinity binding to complementary sequences. LNAs are synthesized from a commercially available sugar, 1,2:5,6-di-O-isopropylidene-a-D-allofuranose. An efficient and simplified procedure is presented for synthesizing a glycol donor that can be used for synthesis of a variety of LNA monomers. Then, as an example, the synthesis of the thymidine analog of LNA from this glycol donor is presented. The protocols give high yields of the desired products and avoid the use of time consuming column chromatography. PMID- 18428895 TI - Cellular delivery of locked nucleic acids (LNAs). AB - Locked nucleic acids (LNAs) are RNA derivatives that have an O-methylene linkage between the 2 and 4 positions of the ribose. This leads to exceptionally high affinity binding to complementary sequences. They are synthesized using standard DNA/RNA synthesis methods, and have a negatively charged backbone that confers good solubility. This unit describes a method for the introduction of LNA oligomers into cells. A support protocol also describes the determination of melting temperatures for LNA oligomers. PMID- 18428896 TI - Solid-phase synthesis of branched oligonucleotides. AB - Branched nucleic acids (bNAs) have been of particular interest since the discovery of RNA forks and lariats as intermediates of nuclear mRNA splicing, as well as multicopy, single-stranded DNA (msDNA). Such molecules contain the inherent trait of vicinal 2',5'- and 3',5'-phosphodiester linkages. bNAs have many potential applications in nucleic acid biochemistry, particularly as tools for studying the substrate specificity of lariat debranching enzymes, and as biological probes for the investigation of branch recognition during pre-mRNA splicing. The protocols described herein allow for the facile solid-phase synthesis of branched DNA and/or RNA oligonucleotides of varying chain length, containing symmetrical or asymmetrical sequences immediate to an RNA branch point. The synthetic methodology utilizes widely adopted phosphoramidite chemistry. Methods for efficient purification of bNAs via anion-exchange HPLC and PAGE are also illustrated. PMID- 18428897 TI - Solid-phase synthesis of 2'-deoxy-2'-fluoro- beta-D-oligoarabinonucleotides (2'F ANA) and their phosphorothioate derivatives. AB - This unit describes the chemical synthesis of 2'-deoxy-2'-fluoro-b-D oligoarabinonucleotides (2'F-ANA), both with phosphodiester and phosphorothioate linkages. The protocols described herein include araF phosphoramidite preparation, assembly on DNA synthesizers, and final deprotection and purification of oligonucleotides. PMID- 18428898 TI - Probing DNA structure with hydroxyl radicals. AB - The hydroxyl radical is a useful probe for studying the shape of the surface of a DNA molecule. Using this technique, fine details of DNA structure can potentially be revealed. This unit describes how to use the hydroxyl radical to generate a random cleavage pattern at the surface of the molecule, separate the broken DNA strands by polyacrylamide gel electrophoresis, and analyze the cleavage pattern to give an image of the surface of the molecule. PMID- 18428899 TI - In vitro selection of RNA aptamers to a small molecule target. AB - This unit describes the selection of aptamers that bind to a small molecule target from a single-stranded RNA pool. Aptamers generated from this type of selection experiment can potentially function as receptors for small molecules for applications ranging from medical diagnostics to environmental monitoring. This unit describes two modes of selection, one by column filtration and one by batch selection. PMID- 18428901 TI - Synthesis, characterization, and application of substituted pyrazolopyrimidine nucleosides. AB - This unit describes, in detail, the preparation of 3-aminopropyl-substituted pyrazolo[3,4-d]pyrimidine analogs of the purines deoxyadenosine (dA) and deoxyguanosine (dG). Phosphoramidite reagents of these so-called aminopropyl-PPA and -PPG nucleosides (AP-PPA and AP-PPG, respectively) allow introduction of amino linkers into internal positions of synthetic DNA strands. Synthesis of suitably protected AP-PPA and AP-PPG phosphoramidites are described. The stepwise alkynylation, hydrogenation, selective protection, and phosphoramidite synthesis is similar for both the PPA and PPG analogs. To demonstrate the application of these reagents, a protocol is given in which a simple DNA strand is synthesized and conjugated to a lipophilic activated ester (dabcyl-SE) to form a stable amide linkage. Utility of this chemistry for preparing internally modified DNA conjugates is discussed. PMID- 18428900 TI - In vitro selection using modified or unnatural nucleotides. AB - The use of modified nucleotides in an RNA or DNA pool to be used for in vitro selection offers many potential advantages, such as the increased stability of the selected nucleic acid against nuclease degradation. This unit provides useful information and protocols for in vitro selection using modified nucleotides. It includes a discussion of when to use modified nucleotides; protocols for preparing a modified RNA pool and verifying its suitability for in vitro selection; and protocols for selecting and amplifying a functionally enriched pool. PMID- 18428902 TI - Synthesis of 1,5-anhydrohexitol building blocks for oligonucleotide synthesis. AB - This unit describes in detail, the optimized preparations of 1,5-anhydrohexitol and the 1,5-anhydrohexitol building blocks for oligonucleotide synthesis (hG, hA, hC, hT). PMID- 18428903 TI - Capillary electrophoresis of DNA. AB - Capillary electrophoresis (CE) is an alternative to conventional slab gel electrophoresis for the separation of DNA fragments. CE offers a number of advantages over slab gel separations in terms of speed, resolution, sensitivity, and data handling. Separation times are generally only a few minutes and the DNA is detected either by UV absorption or by fluorescent labeling. The quantity of DNA required for separation is in the nanogram range. Single-base resolution can be obtained on fragments up to several hundred base pairs. In the presence of appropriate standards, fragments can be accurately sized based on relative electrophoretic mobility. A protocol for the analysis of synthetic oligonucleotides in a flowable matrix is described in this unit. PMID- 18428904 TI - Probing RNA structural dynamics and function by fluorescence resonance energy transfer (FRET). AB - Biological function of RNA is often mediated by cyclic switching between several (meta-)stable arrangements of tertiary structure. Fluorophore labeling of RNA offers a unique view into these folding and conformational switching events, since a fluorescence signal is sensitive to its molecular environment and can be continuously monitored in real time to produce kinetic rate information. This unit focuses on the practical implications of using fluorescence resonance energy transfer (FRET) to probe RNA structural dynamics and function. FRET is a particularly powerful fluorescence technique since, in addition to kinetic data, it provides insights into the structural basis of a conformational rearrangement. Protocols describe how to postsynthetically label RNA for FRET and how to acquire and analyze FRET data. Support protocols describe methods for deprotecting synthetic RNA and for purifying RNA by gel electrophoresis and HPLC. Considerations for selecting appropriate RNA, fluorophores, and labeling strategies are discussed in detail in the commentary. PMID- 18428905 TI - 3-(N-tert-butylcarboxamido)-1-propyl and 4-oxopentyl groups for phosphate/thiophosphate protection in oligodeoxyribonucleotide synthesis. AB - This unit provides procedures for the preparation of deoxyribonucleoside phosphoramidites and appropriate phosphordiamidite precursors with P(III) protecting groups different than the standard 2-cyanoethyl group. Specifically, these phosphoramidites are functionalized with the 3-(N-tert-butylcarboxamido)-1 propyl or 4-oxopentyl groups. The usefulness of these novel deoxyribonucleoside phosphoramidites in the solid-phase synthesis of a 20-mer DNA oligonucleotide and its phosphorothioated analog is demonstrated. It is also shown that removal of the 3-(N-tert-butylcarboxamido)-1-propyl phosphate/thiophosphate-protecting group from these oligonucleotides is rapidly effected under thermolytic conditions at neutral pH, whereas the 4-oxopentyl group is preferably removed by treatment with pressurized ammonia gas or concentrated ammonium hydroxide at ambient temperature. These detailed methods constitute an economical and alkylation-free approach to large-scale preparations of therapeutic oligonucleotides. PMID- 18428906 TI - Chemistry of CpG DNA. AB - The vertebrate immune system can recognize specific pathogen-associated molecular patterns in invading microorganisms, including the unmethylated CpG dinucleotide. This unit discusses the receptors that recognize CpG motifs and important aspects of the sequence context of CpG motifs to the end of understanding and designing CpG DNA for therapeutic purposes. PMID- 18428907 TI - Synthesis of phosphorothioate oligonucleotides with stereodefined phosphorothioate linkages. AB - A method for solid-phase synthesis of stereodefined PS-oligos via an oxathiaphospholane approach using pure P-diastereomers of nucleoside oxathiaphospholane monomers is described. The oxathiaphospholane monomers are synthesized by phosphitylation of 5'-O-DMTr-N-protected deoxyribonucleosides with 2-chloro-spiro-4,4-pentamethylene-1,3,2-oxathiaphospholane followed by sulfurization. The procedure is general and may be applied to other analogs, depending on the aldehyde (or mercaptoalcohol) used. Starting from an 18O-labeled mercaptoalcohol, the corresponding 18O-labeled phosphitylating reagent and nucleoside monomers can be obtained and used for synthesis of labeled stereodefined PS-oligos, which are useful for studying mechanisms of enzymatic reactions. Details are provided for chromatographic separation of the 5'-O-DMTr-N protected-deoxyribonucleoside-3'-O-(2-thio-spiro-4,4-pentamethylene-1,3,2 oxathiaphospholane)s into their P-diastereomers, and for manual solid-phase synthesis of PS-oligos. Oxidation of 5'-O-DMTr-N-protected-deoxyribonucleoside-3' O-(2-thio-spiro-4,4-pentamethylene-1,3,2-oxathiaphospholane)s with selenium dioxide yields their 2-oxo-analogs, which are suitable either for elongation of stereodefined PS-oligos with segments consisting of unmodified nucleotide units possessing phosphate internucleotide linkages, or for generating isotopomeric 18O labeled PO-oligos of predetermined P-chirality. PMID- 18428908 TI - Synthesis of oligonucleotide conjugates via aqueous Diels-Alder cycloaddition. AB - The conjugation of maleimide reporter groups to 5'-diene-modified oligonucleotides via aqueous Diels-Alder cycloaddition is described. Detailed basic protocols are provided for the preparation of a diene-amidite (5'-diene modifier), for the attachment of a diene group to the 5'-terminus of oligonucleotides, and for the conjugation of maleimide reporter groups to diene modified oligonucleotides. PMID- 18428909 TI - 5'-Iodination of solid-phase-linked oligodeoxyribonucleotides. AB - 5'-Iodinated oligodeoxyribonucleotides readily react with 3'-phosphorothioated DNA in the presence of a complementary template to yield a conjugate that is identical to natural DNA in every respect except that one oxygen atom in the phosphodiester backbone is replaced by a sulfur atom. The 5'-iodo group is easily converted to a variety of other functional groups and will quickly react with thiol-containing labels to yield stable thioether conjugates. This unit presents manual and automated procedures for converting the 5'-hydroxyl of protected CPG bound oligodeoxyribonucleotides to an iodo group and for releasing and purifying the products. PMID- 18428910 TI - Reversible biotinylation of the 5'-terminus of oligodeoxyribonucleotides and its application in affinity purification. AB - The preparation of two reversible biotinylation phosphoramidites and their application in labeling and affinity purification of synthetic oligodeoxyribonucleotides will be described. In both cases, the biotin is linked to the 5'-terminus of DNA through a diisopropyl silyl acetal functionality. This linkage is completely stable under certain postsynthetic cleavage/deprotection conditions, but can be readily broken by fluoride ions, releasing unmodified 5' OH and 5'-phosphate DNA, respectively. To demonstrate the use of these reversible biotinylation methods, crude DNA was incubated with NeutrAvidin-coated microspheres, full-length biotinylated DNA was efficiently attached to the solid phase, and nonbiotinylated failure sequences and other impurities were readily removed by washing with buffer. Cleavage of the silyl acetal linkage afforded high-quality, full-length, unmodified 5'-OH and 5'-phosphate DNA, respectively, depending on which of the two phosphoramidites was used. It is anticipated that this method will find applications in areas that require efficient isolation of DNA from a complex mixture. PMID- 18428911 TI - Synthesis of endcap dimethoxytrityl phosphoramidites for endcapped oligonucleotides. AB - Endcaps may be either aromatic or aliphatic molecules that specifically cross link the 5' end of one strand with the 3' end of the complementary strand in a DNA duplex. Endcaps may be viewed as a replacement of the loop region nucleotides of a DNA hairpin, with the added advantage of increased thermal stability. An endcap is incorporated into the sequence during oligonucleotide synthesis. Three endcaps are described in this unit. The naphthalene diimide endcap prefers to base stack with GC base pairs. The terthiophene endcap has higher lipophilicity than the naphthalene diimide endcap and provides higher stability when stacked over an AT base pair. The 2,2'-oxydiacetamide endcap provides lower enhancement in stability but a more rigid and well-defined structure than the oligo(ethylene glycol) endcaps. Synthesis of endcapped oligonucleotides can be carried out using standard automated synthesis protocols with only minor modifications. PMID- 18428912 TI - Engineering terminal disulfide bonds into DNA. AB - This protocol presents a simple and general means of modifying nucleic acids with disulfide cross-links. These cross-links serve as powerful tools to probe the structure, dynamics, thermodynamics, folding, and function of DNA and RNA, much in the way that cystine cross-links have been used to study proteins. The chemistry described has been used to synthesize disulfide-cross-linked hairpins and duplexes, higher-order structures such as triplexes, non-ground-state conformations, and tRNAs. since the cross-links form quantitatively by mild air oxidation and do not purturb either secondary or tertiary structure, this modification should prove quite useful for the study of nucleic acids. PMID- 18428914 TI - Characterization of DNA structures by circular dichroism. AB - Circular dichroism (CD) of nucleic acids is commonly used to provide a signature for a given secondary structure. CD is particularly powerful for monitoring structural changes resulting from changes in environmental conditions such as temperature, ionic strength, and pH. A number of commercially available instruments are available for the measurement of CD spectra. Since the details of instrumental operation differ for each machine, this unit provides a descriptive guide, rather than a strict protocol. It describes considerations that the experimentalist will face when attempting to examine nucleic acid structures by CD spectropolarimetry. Specific details are also given for cell filling and cleaning, and for instrumental calibration. Finally, several examples of CD spectra are given to provide useful reference points for a variety of nucleic acid structures, ranging from single-stranded to triplex and quadruplex forms. PMID- 18428913 TI - Probing RNA structure and metal-binding sites using terbium(III) footprinting. AB - The function of an RNA molecule is determined by its overall secondary and tertiary structure. The tertiary structure is facilitated and stabilized by the interaction with metal ions. The current chapter offers a detailed protocol on the use of the lanthanide metal ion terbium(III) as a powerful probe of RNA structure and metal-binding properties. When incubating RNA with low (micromolar) concentrations of terbium(III), specific backbone scission by partially deprotonated aqueous terbium(III) complexes can be used to detect high-affinity metal-binding sites, while incubation with high (millimolar) terbium(III) concentrations cleaves the RNA backbone preferentially at structurally accessible regions, providing a footprint of the RNA secondary and tertiary structure. PMID- 18428915 TI - A competition dialysis assay for the study of structure-selective ligand binding to nucleic acids. AB - Unique DNA structures represent potential targets for small molecules, and provide a promising new avenue for drug development. However, attempts to rationally design small molecules that bind selectively to a particular DNA structure have been hampered by the lack of a rapid and convenient assay for structural selectivity. Determination of structure-selective ligand binding using competition dialysis is described in this unit. The competition dialysis assay is simple, straightforward, and rapid once stock solutions of the nucleic acid structures of interest have been prepared as described. PMID- 18428917 TI - Selected abbreviations used in this manual. AB - This APPENDIX provides commonly used abbreviations for many terms used in in this manual. PMID- 18428916 TI - Chemistry of minor groove binder-oligonucleotide conjugates. AB - Various types of minor groove binders have been attached to synthetic oligodeoxynucleotides, and the interactions of these conjugates (MB-ODNs) with DNA are reviewed here. MB-ODNs have enhanced DNA affinity and have improved the hybridization properties of sequence-specific DNA probes. Short MB-ODNs hybridize with ssDNA to give more stable DNA duplexes than unmodified ODNs with similar lengths. Mismatch discrimination of short MB-ODNs is enhanced in comparison to longer unmodified ODNs. The stronger binding of MB-ODNs allows for more stringent hybridization conditions to be used in DNA probe-based assays. MB-ODNs are especially useful in quantitative "real-time" PCR assays since they bind efficiently during the high-temperature primer extension cycle. The synthesis and biophysical chemistry of MB-ODN conjugates are reviewed here. Four published structural classes of MB-ODNs and their various dsDNA binding modes are discussed, and the well-characterized DPI3-type MB-ODNs and their interactions with ssDNA target strands are described in detail. PMID- 18428918 TI - Site-specific fluorescent labeling of large RNAs with pyrene. AB - Pyrene is a useful chromophore for monitoring the tertiary structure and folding of large RNAs. This unit describes the general preparation of a large RNA (>80 nucleotides in length) that has been site-specifically modified with pyrene at the 2'-position of an individual internal nucleotide. A protocol is provided for derivatizing a 2'-amino-RNA oligonucleotide with a suitably activated pyrene reagent. This pyrene-labeled oligonucleotide is then assembled with other RNA(s) either by covalent ligation or by noncovalent hybridization to form a full-length structured RNA, which may then be studied by equilibrium and stopped-flow fluorescence spectroscopy. PMID- 18428919 TI - Synthesis of 5'-O-phosphoramidites with a photolabile 3'-O-protecting group. AB - This unit describes the chemical synthesis of phosphoramidite building blocks that carry a protecting group at the 3' position. These inversely oriented synthons expose a 2-(2-nitrophenyl)propoxycarbonyl (NPPOC) group as the photolabile protecting group of choice. Among other applications, the building blocks can be employed for light-controlled in situ synthesis of DNA microarrays, producing arrayed oligonucleotides that are attached to the support via their 5' ends, leaving their 3' termini available to act as substrates for polymerases. PMID- 18428920 TI - Derivatization of glass and polypropylene surfaces. AB - This unit describes the derivatization of solid support media, particularly glass and polypropylene, with a linker system by an iterative process that forms a dendrimeric structure, thereby increasing stepwise the loading capacity of the surface. The procedure permits the production of various types of linkers whose characteristics can be tailored to the requirements of the eventual applications. The specific modifications described here are used either for attaching prefabricated DNA oligonucleotides, PCR products, and peptide nucleic acid (PNA) oligomers, or for in situ synthesis of DNA microarrays. PMID- 18428921 TI - Overview of the synthesis of nucleoside phosphates and polyphosphates. AB - This overview summarizes methodology used for the synthesis of nucleoside mono-, di-, and triphosphates. Selected techniques such as the Mitsunobu reaction, displacement reactions involving nucleoside 5'-tosylates, "anion-exchange" techniques, and phosphoramidite and phosphoramidate methodologies are highlighted. The chemistry of phosphorylation is detailed with respect to advantages and limitations under various conditions. Applicability of the methods toward the synthesis of analogs such as imidophosphates, phosphorothioates, and radiolabeled nucleotides is also addressed. PMID- 18428922 TI - Chemoenzymatic preparation of nucleoside triphosphates. AB - The design and synthesis of alternative nucleoside triphosphate substrates for DNA and RNA polymerases holds continued promise to create biochemical probes and precursors for synthesis of nucleic acid mimics. The azole carboxamide nucleotides are of particular interest, as they display multiple conformations in the context of DNA replication. An efficient chemoenzymatic preparation of azole carboxamide deoxyribo- and ribonucleoside triphosphates is presented. Nucleoside diphosphate is prepared from nucleoside 5'-O-tosylate by displacement with tris(tetra-n-butylammonium) pyrophosphate. Enzymatic phosphorylation of the azole carboxamide deoxyribonucleoside diphosphate to its triphosphate is based on ATP as the phosphate donor and nucleoside diphosphate kinase as the catalyst, coupled with phospho(enol)pyruvate (PEP) and pyruvate kinase as an ATP regeneration system. Enzymatic phosphorylation of the azole carboxamide ribonucleoside diphosphate requires PEP as the phosphate donor and pyruvate kinase as the catalyst. The optimized purification uses boronate affinity gel to yield highly purified nucleoside triphosphate. PMID- 18428923 TI - Synthesis and polymerase incorporation of 5'-amino-2',5'-dideoxy-5'-N triphosphate nucleotides. AB - This unit presents synthetic procedures for the preparation of 5'-amino-2',5' dideoxy analogs of adenosine, cytidine, guanosine, and thymidine, as well as corresponding 5'-N-triphosphate nucleotides, using commercially available reagents. The modified nucleosides are prepared in high yields from naturally occurring 2'-deoxynucleosides using robust chemical reactions including tosylation, azide exchange, and the Staudinger reaction. Efficient conversion of these 5'-amino nucleosides to corresponding 5'-N-triphosphate nucleotides is achieved through a one-step reaction with trimetaphosphate in Tris-buffered aqueous solution. The 5'-amino modification renders these nucleoside and nucleotide analogs markedly increased reactivity, which is useful for a variety of biochemical, pharmaceutical, and genomic applications. Also included in this unit are protocols for polymerase incorporation of the 5'-amino nucleotides, either partially or completely replacing their naturally occurring counterparts, and subsequent sequence-specific cleavage at the modified nucleotides under mildly acidic conditions. PMID- 18428924 TI - Preparation of 5'-silyl-2'-orthoester ribonucleosides for use in oligoribonucleotide synthesis. AB - The recent discovery that small interfering RNAs (siRNAs) induce gene suppression in mammalian cells has sparked tremendous interest in using siRNA-based assays and high-throughput screens to study gene function. As a result, research programs at leading academic and commercial institutions have become a substantial and rapidly growing market for synthetic RNA. Important considerations in synthesizing RNA for biological gene function studies are sequence integrity, purity, scalability, and resistance to nucleases; ease of chemical modification, deprotection, and handling; and cost. Of the well established RNA synthesis methods, 2'-ACE chemistry is the only one that meets all of these criteria. 2'-ACE technology employs a unique class of silyl ethers to protect the 5'-hydroxyl, in combination with an acid-labile orthoester protecting group on the 2'-hydroxyl (2'-ACE). 2'-ACE-protected phosphoramidite monomers are joined using standard solid-phase technology to achieve RNA synthesis at efficiencies rivaling those for DNA. This unit describes the synthesis of standard 5'-silyl-2'-ACE-protected phosphoramidites. PMID- 18428925 TI - DNA synthesis without base protection. AB - DNA synthesis can be achieved by using O-selective methods for internucleotide bond formation. This greatly simplifies the synthesis of oligodeoxyribonucleotides by eliminating the need for nucleobase protection and deprotection steps. This unit describes strategies that can be used for DNA synthesis without base protection. The discussion includes synthesis of phosphoramidite and H-phosphonate monomers, solid-phase assembly by the phosphoramidite and H-phosphonate methods, and future prospects for DNA synthesis using N-unprotected approaches. PMID- 18428926 TI - The 4-methylthio-1-butyl group for phosphate/thiophosphate protection in oligodeoxyribonucleotide synthesis. AB - The detailed preparation of deoxyribonucleoside phosphoramidites functionalized with a 4-methylthio-1-butyl group for P(III) protection is described, along with the incorporation of these phosphoramidites into DNA oligonucleotides via solid phase techniques. The versatility of the thermolabile 4-methylthio-1-butyl phosphate/thiophosphate-protecting group is exemplified through its facile removal from oligonucleotides under neutral conditions or under standard basic conditions. The sulfonium salt that is produced during the thermolytic deprotection of oligonucleotides did not alter DNA nucleobases or desulfurize phosphorothioate diesters to a significant extent. PMID- 18428927 TI - Synthesis of oligodeoxyribo- and oligoribonucleotides according to the H phosphonate method. AB - Oligonucleotides can be synthesized by condensing a protected nucleoside H phosphonate monoester with a second nucleoside in the presence of a coupling agent to produce a dinucleoside H-phosphonate diester. This can then be converted to a dinucleoside phosphate or to a backbone-modified analog such as a phosphorothioate or phosphoramidite. This unit discusses four alternative methods for synthesizing nucleoside H-phosphonate monoesters. The methods are efficient and experimentally simple, and use readily available reagents. The unit describes the activation of the monoesters, as well as competing acylation and other potential side reactions. PMID- 18428928 TI - Uridine 2'-carbamates: facile tools for oligonucleotide 2'-functionalization. AB - A facile method for preparation of uridine 2'-carbamate derivatives based on reaction of 3',5'-disilyl-protected uridine with 1,1'-carbonyldiimidazole followed by treatment with an aliphatic amine is presented. A phosphoramidite monomer suitable for automated oligonucleotide synthesis is obtained in a few steps. The compounds are useful for the introduction of various labels and modifications into an oligonucleotide chain. Although 2'-carbamate modification is somewhat destabilizing for DNA-DNA and DNA-RNA duplexes, it is suitable for the direction of ligands into the minor groove of duplexes or at non-base-paired sites (e.g., loops and bulges) of oligonucleotides. Pyrene-modified oligonucleotide 2'-carbamates show a considerable increase in fluorescence intensity upon hybridization to a complementary RNA (but not DNA). PMID- 18428929 TI - Synthesis of oligoribonucleotides containing N6-alkyladenosine and 2-methylthio N6-alkyladenosine. AB - The N(6)-alkyladenosines and 2-methylthio-N(6)-alkyladenosines are the most common modified adenosine nucleosides, and transfer ribonucleic acids (tRNA) are particularly rich in these modified nucleosides. They are present at position 37 of the anticodon arm, and the contributions of these hypermodified nucleosides to codon-anticodon interactions as well as to translation are significant, although they are not fully understood. This unit describes a new chemical synthesis method for oligoribonucleotides containing N(6)-alkyladenosines and 2-methylthio N(6)-alkyladenosines via postsynthetic modifications of precursor oligoribonucleotides. To obtain oligoribonucleotides containing N(6) alkyladenosines, a precursor oligoribonucleotide carrying 6-methylthiopurine riboside residues was used, whereas for the synthesis of oligoribonucleotides containing 2-methylthio-N(6)-alkyladenosines, a precursor oligoribonucleotide carrying the 2-methylthio-6-chloropurine riboside was applied. This allowed synthesis of modified oligoribonucleotides containing naturally occurring modified nucleosides such as N(6)-isopentenyladenosine (i(6)A), N(6) methyladenosine (m(6)A), 2-methylthio-N(6)-isopentenyladenosine (ms(2)i(6)A), and 2-methylthio-N(6)-methyladenosine (ms(2)m(6)A), as well as several unnaturally modified adenosine derivatives. PMID- 18428930 TI - Oligodeoxyribonucleotide analogs functionalized with phosphonoacetate and thiophosphonoacetate diesters. AB - Oligodeoxyribonucleotides with phosphonoacetate or thiophosphonoacetate internucleotide linkages can be made in high yield by solid-phase synthesis and possess many advantages. They are highly stable to nucleases, water-soluble, and anionic at neutral pH. They form stable duplexes with DNA and RNA, and stimulate RNase H degradation of complementary RNA. The preparation of the N,N (diisopropylamino)phosphinyl acetate monomers from standard protected nucleosides is described here, followed by the synthesis of phosphonoacetate and thiophosphonoate oligodeoxyribonucleotides, as well as chimeric oligomers that have these modified linkages in combination with natural or phosphorothioate linkages. Purification and characterization of these oligomers is also presented. PMID- 18428932 TI - Synthesis and properties of 7-substituted 7-deazapurine (pyrrolo[2,3 d]pyrimidine) 2'-deoxyribonucleosides. AB - 7-Substituted 7-deazapurine (pyrrolo[2,3-d]pyrimidine) 2'-deoxyribonucleosides are synthesized by stereoselective nucleobase anion glycosylation. The introduction of a halogen at C7 is performed regioselectively either on the nucleobase or on the nucleoside. The pK(a) values of a series of 7-deazapurine 2' deoxyribonucleosides are provided, and fluorescence properties are also discussed. PMID- 18428931 TI - Probing RNA structure and function by nucleotide analog interference mapping. AB - Nucleotide analog interference mapping (NAIM) can be used to simultaneously, yet individually, identify structurally or catalytically important functional groups within an RNA molecule. Phosphorothioate-tagged nucleotides and nucleotide analogs are randomly incorporated into an RNA of interest by in vitro transcription. The phosphorothioate tag marks the site of substitution and identifies sites at which the modification affects the structure or function of the RNA molecule. This technique has been expanded to include identification of hydrogen bonding pairs (NAIS), ionizable functional groups, metal ion ligands, and the energetics of protein binding (QNAIM). The analogs, techniques, and data analysis used in NAIM are described here. PMID- 18428933 TI - Reduction of ribonucleosides to 2'-deoxyribonucleosides. AB - Ribonucleosides are converted into 2'-deoxyribonucleosides in good yields by a four-step procedure. Selective protection of the 3'- and 5'-hydroxyl groups with 1,3-dichloro-1,1,3,3-tetraisopropyl-1,3-disiloxane is followed by functionalization of the 2'-hydroxyl group with phenoxythiocarbonyl chloride. Free radical-mediated reductive C2'-O2' bond cleavage of these 3',5'-O-TPDS-2'-O PTC-nucleoside derivatives with tributyltin hydride, followed by removal of the silyl protecting group with tetrabutylammonium fluoride, provides the 2' deoxyribonucleosides. Adenosine, cytidine, guanosine, and uridine are converted into dA, dC, dG, and dU in overall yields of 60% to 80%. Use of tributyltin deuteride in the reductive cleavage step gives 2'-deuterio-2'-deoxyadenosine in 81% yield from adenosine with >85% retention of configuration at C2'. Application of this four-step protocol with nucleoside analogs is straightforward. PMID- 18428934 TI - DNA microarray preparation by light-controlled in situ synthesis. AB - This unit describes the in situ synthesis of DNA microarrays using a light controlled process. In a highly parallel fashion, multiple oligonucleotide probes are created from scratch on a glass support using phosphoramidite building blocks carrying a photolabile protecting group. Spatial resolution is achieved by carefully controlling the illumination of defined spots on the glass substrate, on which the oligonucleotide probes are synthesized using straightforward phosphoramidite chemistry. PMID- 18428935 TI - Preparation of alpha-Oxo semicarbazone oligonucleotide microarrays. AB - This unit describes the preparation of alpha-oxo aldehyde functionalized oligodeoxynucleotides, the preparation and characterization of semicarbazide glass slides, and the fabrication of alpha-oxo semicarbazone microarrays by site specific ligation of alpha-oxo-aldehyde oligodeoxynucleotides to the semicarbazide glass slides. The alpha-oxo aldehyde group COCHO is extensively used in ligation chemistry for the preparation of large molecular constructs. It is stable toward air oxidation and mainly present in aqueous solution in the hydrated form COC(OH)(2). It reacts efficiently with hydrazine derivatives, in particular, with the semicarbazide group. The reaction occurs spontaneously in water at pH 5.5. Site-specific immobilization of glyoxylyl oligodeoxynucleotides on semicarbazide glass slides allows the preparation of high-quality microarrays that can be used directly in hybridization experiments. PMID- 18428936 TI - Synthesis of covalent oligonucleotide-streptavidin conjugates and their application in DNA-directed immobilization (DDI) of proteins. AB - Semisynthetic DNA-streptavidin conjugates are synthesized by covalent coupling of thiol-modified DNA oligonucleotides and streptavidin (STV). The resulting conjugates have binding capacities for four equivalents of biotin and a complementary nucleic acid sequence. The conjugates are purified to homogeneity by ultrafiltration and chromatography, and are characterized by spectrophotometry and gel electrophoresis. Subsequently, the conjugates are applied as molecular connectors in the DNA-directed immobilization (DDI) of biotinylated antibodies using DNA microarrays as immobilization matrices. The results are protein microarrays that can be used for the multiplexed detection of various antigens. PMID- 18428937 TI - Synthesis of acyclic analogs of adenosine. AB - This unit describes preparation of two groups of chiral acyclic nucleosides that are N-alkyl derivatives of adenine with hydroxyl group(s) on the alkyl chain. As an example of the neutral group, the synthesis is presented for (S) (2,3,dihydroxypropyl)adenine (DHPA), a compound with multiple biological effects in proliferating systems. Its synthesis consists of alkylation of adenine with a suitable chiral synthon based on 1,2-O-isopropylideneglycerol. Another group of structurally related biologically active compounds are the 9-alkyladenines bearing both hydroxyl and carboxyl functions. Large-scale cyanohydrin reaction of the adenine-containing acetaldehyde intermediate yields 3-(adenine-9-yl)-2 hydroxypropanoic acid (AHPA). Lastly, one of the alternative syntheses of its homolog, the nucleoside antibiotic eritadenine (with a D-erythro-2,3 dihydroxybutanoic acid side-chain) is described starting from d-ribose. Procedures for preparation of all intermediates as well as the purification of the resulting crude material are described in detail. All these adenine derivatives inhibit SAM-dependent methylation reactions via inhibition of SAH hydrolase. PMID- 18428938 TI - Synthesis of acyclic nucleoside phosphonates. AB - Acyclic nucleoside phosphonates (ANPs) are important biologically active nucleotide analogs. They contain an isopolar phosphonomethyl function linked to the hydroxyl group of an acyclic side-chain via an undegradable ether group. Though their most important activity is antiviral, some exhibit cytostatic or antiprotozoic effects. The three most important groups of ANP are presented here as synthetic procedures for a large laboratory scale. Synthesis follows three principles: (1) introduction of a protected phosphonomethyl group to the hydroxyl on an appropriate alkyl side-chain of an acyclic nucleoside, (2) alkylation of the heterocyclic base by a synthon with all characteristic features of the future phoshonate-bearing side-chain, or (3) transformation of a reactive group at the heterocyclic base. The last step in all these cases is removal of the phosphonate esters. Preparation methods are described in detail for PMEA, PMEG, PMEDAP and its N(6)-cyclopropyl derivative, (R)-PMPA, and (S)-HPMPA, as well as all intermediates and synthons. PMID- 18428939 TI - Enzymatic regioselective levulinylation of 2'-deoxyribonucleosides and 2'-o methylribonucleosides. AB - The levulinyl-protected nucleosides are key building blocks for the solution phase synthesis of oligonucleotides. Short and efficient syntheses of 3'- and 5' O-levulinylated 2'-deoxyribonucleosides and 2'-O-methylribonucleosides have been developed from the corresponding nucleosides by enzyme-catalyzed regioselective acylation in organic solvents or from 3',5'-di-O-levulinyl derivatives by regioselective enzymatic hydrolysis. Lipase-mediated levulinylation of various nucleosides has been accomplished with acetonoxime levulinate as an acyl donor. Use of immobilized Pseudomonas cepacia lipase (PSL-C) has furnished 3'-O levulinylated 2'-deoxyribonucleosides in excellent yields. Similarly, Candida antarctica lipase B (CAL-B) has provided 5'-O-levulinylated nucleosides in high yields. 3'-O-Levulinylated 2'-deoxyribonucleosides and 2'-O-methylribonucleosides were prepared via selective hydrolysis of 3',5'-di-O-levulinate esters using CAL B. PMID- 18428940 TI - Base-modified oligodeoxyribonucleotides: using pyrrolo[2,3-d]pyrimidines to replace purines. AB - Phosphoramidites are first synthesized from suitably protected 7-deazapurine 2' deoxyribonuclosides that have halogen or alkynyl subsituents at the 7 position. Oligonucleotides containing 7-deazapurines (pyrrolo[2,3-d]pyrimidines) in place of the canonical nucleobases are then prepared by solid-phase synthesis. These oligonucleotides are subsequently allowed to hybridize, and the stabilities of the resulting duplexes are determined. Comparison of T(m) values indicates that the presence of 7-halo or 7-alkynyl substituents increases duplex stability relative to duplexes involving oligonucleotides that contain the corresponding parent purines. PMID- 18428942 TI - Large-scale preparation of conjugated oligonucleoside phosphorothioates by the high-efficiency liquid-phase (HELP) method. AB - A new process for the preparation of large amounts of thioated oligonucleotides in a quasi-classical solution condition is described. This method takes advantage of the use of polyethylene glycol as a soluble, inert support during synthesis. The easy purification of intermediates from a moderate excess of reagents allows very high coupling yields and, consequently, efficient production of the long oligonucleotide sequences required for pharmacological applications. The reaction of properly protected and activated oligonucleotides with high-molecular-weight polyethylene glycols allows a convenient procedure for the postsynthetic conjugation of those biopolymers in solution. The oligonucleotides are modified at the 5' terminus using a liquid-phase procedure with a linker carrying a terminal primary amino group to enhance its nucleophilic reactivity. The use of N, N'-disuccinimidyl carbonate for the activation of the terminal OH groups of the PEG was preferred. PMID- 18428941 TI - An aminooxy-functionalized non-nucleosidic phosphoramidite for the construction of multiantennary oligonucleotide glycoconjugates on a solid support. AB - In this unit, a method is described that allows construction of multiantennary oligonucleotide glycoconjugates on a solid support. A bis(hydroxymethyl)malondiamide-based phosphoramidite that contains two phthaloyl protected aminooxy groups compatible with normal chain assembly is prepared. The aminooxy functions can be deblocked with a hydrazinium acetate treatment and subsequently oximated on-support with fully acetylated 4-oxobutyl alpha-D mannopyranoside. The resulting reagent is then used to prepare a conjugate containing two non-nucleosidic building blocks (i.e., four alpha-D-mannopyranosyl units) close to the 5' terminus of the oligonucleotide. PMID- 18428943 TI - A fluorescent intercalator displacement assay for establishing DNA binding selectivity and affinity. AB - A protocol for a fluorescent intercalator displacement (FID) assay useful for establishing DNA binding selectivity, affinity, stoichiometry, and binding site size, and for distinguishing modes of DNA binding is presented. PMID- 18428944 TI - Synthesis of fluorinated nucleosides. AB - Two practical synthetic approaches to the production of lodenosine [FddA, 9-(2,3 dideoxy-2-fluoro-beta-D-threo-pentofuranosyl)adenine] via 6-chloropurine riboside or 6-chloropurine 3'-deoxyriboside are described. The reaction sequence contains new fluorination methods and new applications of radical reduction. The reagents and reaction conditions of each step have been carefully selected to ensure robustness and safety. PMID- 18428945 TI - Sequencing oligonucleotides by enrichment of coupling failures using matrix assisted laser desorption/ionization time-of-flight mass spectrometry. AB - A technique for sequencing oligonucleotides using matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry is described. Coupling failures are extracted from the full-length 5'-O-DMTr-oligonucleotide in a crude synthesis using C18 purification cartridges. Enhanced signal and resolution of the failure ions are demonstrated during MALDI-TOF analysis. Sequencing information is confirmed by the mass difference between coupling failures to identify a particular base or structural modification. This procedure eliminates difficulties associated with other mass spectrometric techniques, such as interpretation of data, oligonucleotide length, and backbone, sugar or terminal alterations. An example of sequencing a 16-mer composed of deoxyribonucleotides, 2'-O-methylribonucleotides, and a non-nucleosidic linker is reported. PMID- 18428946 TI - Mass determination of phosphoramidites. AB - Nucleoside phosphoramidites are the most widely used building blocks in contemporary solid-phase synthesis of oligonucleotides. The accurate molecular weight measurements of such molecules, which are acid-labile compounds, may be easily determined by mass spectrometry using a matrix system, triethanolamine/NaCl, on a liquid secondary ion mass spectrometer (LSIMS) or fast atom bombardment (FAB) MS equipped with a double-focusing mass spectrometer. The present method rapidly and easily measures the accurate molecular weights of various phosphoramidites as adduct ions [M+Na]+ with an average mass error smaller than 0.4 ppm, allowing determination of the formulas of the phosphoramidites in place of elemental analysis. Further, it was found that intensities of molecular-related ions could be enhanced to the highest degree by adjustment of the molar ratio of phosphoramidite and NaCl, fixing the amount of triethanolamine on LSIMS, making the present method a powerful tool for identification of phosphoramidites by mass spectrometry. PMID- 18428947 TI - Nucleoside-5'-phosphoimidazolides: reagents for facile synthesis of dinucleoside pyrophosphates. AB - A facile method is presented for preparation of dinucleoside pyrophosphate derivatives based on reaction of a nucleoside 5'-monophosphate with carbonyldiimidazole followed by treatment of the resulting nucleoside 5' phosphoimidazolide with a nucleoside 5'-phosphate. This method is suitable for preparation of pyrophosphates analogous to NAD, FAD, and related natural pyrophosphates. The resulting compounds are useful for mechanistic studies of enzymes that use natural pyrophosphates as co-factors or substrates, and in development of inhibitors that have potential applications as therapeutic agents. PMID- 18428948 TI - Synthesis of methylenebis(phosphonate) analogs of dinucleotide pyrophosphates. AB - A facile method is described for preparation of nonsymmetrical P1,P2 methylenebis(phosphonate) diesters based on the 31P-NMR-controlled reaction of methylenebis(phosphonate) monoesters with diisopropylcarbodiimide, resulting in the formation of the intermediate P1,P4-disubstituted bicyclic trisanhydride. This intermediate, after treatment with an another nucleoside, carbohydrate, or alcohol followed by hydrolysis, is converted into the corresponding methylenebis(phosphonate) diester. An analog of a natural dinucleotide pyrophosphate can be obtained when a nucleoside 5'-methylenebis(phosphonate) is coupled with another nucleoside. This method is suitable for preparation of metabolically stable (resistant to phosphodiesterase cleavage) analogs of NAD, FAD, and related natural pyrophosphates. The resulting compounds are useful for mechanistic studies of enzymes that use the natural pyrophosphates as cofactors or substrates, and in development of inhibitors that have potential applications as therapeutic agents. PMID- 18428949 TI - Chemical phosphorylation of deoxyribonucleosides and thermolytic DNA oligonucleotides. AB - The phosphorylating reagent bis[S-(4,4'-dimethoxytrityl)-2-mercaptoethyl]-N,N diisopropylphosphoramidite is prepared in three steps from commercial methyl thioglycolate and diisopropylphosphoramidous dichloride. The phosphorylating reagent has been used successfully in the solid-phase synthesis of deoxyribonucleoside 5'-/3'-phosphate or -thiophosphate monoesters and oligonucleotide 5'-phosphate/-thiophosphate monoesters. Bis[S-(4,4' dimethoxytrityl)-2-mercaptoethyl]-N,N-diisopropylphosphoramidite has also been employed in the construction of a thermolytic dinucleotide prodrug model to evaluate the ability of the reagent to produce thermosentive oligonucleotide prodrugs under mild temperature conditions ( approximately 25 degrees C) for potential therapeutic applications. PMID- 18428950 TI - Synthesis of beta-L-2'-deoxythymidine (L-dT). AB - The "unnatural" l-nucleoside beta-l-2'-deoxythymidine (L-dT) is a potent, specific, and selective inhibitor of the replication of hepatitis B virus (HBV), which is currently in Phase III clinical trials. This unit describes, in detail, a semi-large-scale synthesis of l-dT. This convenient methodology produces l-dT in six steps starting with l-ribose and ending with a satisfactory overall yield of l-dT, and may be applied to other 2'-deoxynucleosides, incorporating different heterocyclic bases. PMID- 18428951 TI - Synthesis of carbovir and abacavir from a carbocyclic precursor. AB - A facile method for the synthesis of a carbocyclic analog of 2',3'-didehydro 2',3'-dideoxy-2-amino-6-chloropurine is presented, starting from the versatile lactam, 2-azabicyclo[2.2.1]hept-5-en-3-one. The corresponding anti-HIV nucleoside analogs, carbovir and its derivative abacavir, are prepared directly from the 6 chloro intermediate in one-step reactions. PMID- 18428952 TI - Synthesis of amino acid phosphoramidate monoesters via H-phosphonate intermediates. AB - Diphenyl phosphite and bis(N,N-diisopropylamino)chlorophosphine are used as phosphitylating reagents to generate H-phosphonate monoesters. These H phosphonate intermediates are subsequently oxidized with iodine to generate the 5'-nucleoside amino acid phosphoramidates. PMID- 18428953 TI - Nucleobase protection with allyloxycarbonyl. AB - This unit describes protocols for preparation of N-allyloxycarbonyl-protected 5' O-(4,4'-dimethoxytrityl)-2'-deoxyribonucleosides and 2'-O-(tert butyldimethylsilyl)-5'-O-(4,4'-dimethoxytrityl)ribonucleosides. These provide useful building blocks not only for synthesis of natural oligonucleotides but also for artificial analogs with chemically sensitive (particularly, base-labile) modified nucleoside bases or internucleotide linkages. These protected nucleosides are stable to conditions used for conversion to the corresponding nucleoside phosphoramidites and subsequent oligonucleotide synthesis, and the N allyloxycarbonyl protecting group can be easily and cleanly removed by an organopalladium-catalyzed reaction under mild, nearly neutral conditions. PMID- 18428954 TI - Nucleoside phosphoramidites containing cleavable linkers. AB - Phosphoramidite reagents (linker phosphoramidites) containing a cleavable 3' ester linkage between the nucleoside and the phosphoramidite group can be used to attach the first nucleoside to a solid-phase support. Inexpensive underivatized supports such as LCAA-CPG can then be used as universal supports for oligonucleotide synthesis. No modifications to synthesis coupling conditions and no 3'-dephosphorylation are required. Only oligonucleotides with terminal 3'-OH ends are produced. Phosphoramidites containing both a succinate and a sulfonyldiethanol linkage are particularly useful and create oligonucleotides with both a 3'-OH and 5'-phosphate. In addition, by using these reagents, one oligonucleotide sequence can be added onto the 5'-end of another (tandem synthesis) to produce a string of multiple oligonucleotides linked end-to-end. Deprotection releases the oligonucleotides from each other to yield a mixture of oligonucleotides. This approach is particularly useful for making pairs of PCR primers or both strands of a double-stranded sequence in a single operation. PMID- 18428955 TI - Microwave-assisted functionalization of solid supports for rapid loading of nucleosides. AB - Ultra-fast and efficient functionalization of solid supports such as controlled pore glass (CPG), amino methyl polystyrene, and Tentagel has been achieved using microwave-assisted procedures. Both amino- and carboxy-terminated supports are easily prepared within minutes, in a reproducible manner, using microwave assisted methodologies. The resulting functionalized supports are efficiently coupled to nucleosides using dimethylformamide as a solvent in conjunction with a specially designed reactor and workstation called LOTUS. Using these improved protocols, CPG with loadings of 75 to 85 micromol/g can be prepared on a large scale within 3 to 4 days starting from native CPG, as opposed to traditional methods that require 10 to 15 days to achieve the same objective. In addition, the methods described here can potentially be employed for rapid functionalization of other solid matrices such as beads, slides, and pins for applications in microarrays or combinatorial chemistry. PMID- 18428956 TI - Solution-phase synthesis of di- and trinucleotides using polymer-supported reagents. AB - A solution-phase phosphoramidite synthesis of short oligonucleotides using solid supported reagents is described. Polyvinyl pyridinium tosylate is employed as the activator in the coupling step between nucleoside-3'-O-phosphoramidites and 5'-OH nucleosides. The resulting dinucleotide phosphite triesters are oxidized or sulfurized with polymer-bound periodate or tetrathionate to phosphotriesters or thiono phosphotriesters, respectively. Removal of the 3'-O-levulinyl protection is accomplished with a polymer-supported hydrazine. In this approach, the excess reagents are removed by filtration, avoiding complicated purifications. All the polymer-bound reagents used in this method result from salt bridge interactions with the polymers. Consequently, the resins are regenerable. PMID- 18428957 TI - DNA synthesis without base protection using the phosphoramidite approach. AB - In the strategy of DNA synthesis without base protection, a recent O-selective phosphorylation procedure known as the "activated phosphite method" is used to generate reactive phosphite intermediates from N-unprotected phosphoramidite building blocks. This new strategy proved to be advantageous over previous strategies because the cleavage of the N-P(III) bond, which can be formed by the significant phosphitylation of the nucleobase amino groups, can be omitted. The activated phosphite method provides an attractive route to high-throughput synthesis of DNA and the synthesis of alkali-labile modified DNAs such as oligonucleotides containing DNA lesions. This unit presents two convenient methods for the synthesis of N-unprotected deoxynucleoside 3'-phosphoramidite units and detailed procedures for automated synthesis of oligodeoxyribonucleotides from these phosphoramidite units. PMID- 18428958 TI - Disulfide conjugation of peptides to oligonucleotides and their analogs. AB - Peptide conjugation of oligonucleotides and their analogs is being studied widely towards improving the delivery of oligonucleotides into cells. Amongst the many possible routes of conjugation, the disulfide linkage has proved to be the most popular. This reversible linkage may have advantages for cell delivery, since it is likely to be cleaved within cells, thus releasing the oligonucleotide cargo. It is straightforward to introduce thiol functionalities into both oligonucleotide and peptide components suitable for disulfide conjugation. However, severe difficulties have been encountered in carrying out conjugations between highly cationic peptides and negatively charged oligonucleotides because of aggregation and precipitation. Presented here are reliable protocols for disulfide conjugation that have been verified for both cationic and hydrophobic peptides as well as oligonucleotides containing deoxyribonucleosides, ribonucleosides, 2'-O-methylribonucleosides, locked nucleic acid (LNA) units, as well as phosphorothioate backbones. Also presented are reliable protocols for disulfide conjugation of peptide nucleic acids (PNAs) with peptides. PMID- 18428959 TI - Methoxyoxalamido chemistry in the synthesis of tethered phosphoramidites and functionalized oligonucleotides. AB - A general approach to phosphoramidites tethered with single and multiple linkers through the use of methoxyoxalamido (MOX) chemistry is described. The approach utilizes readily available and inexpensive primary aliphatic amino alcohols and diamines to produce a rich and diverse variety of tethered phosphoramidites. Furthermore, the use of MOX chemistry in a modular fashion enables fairly rapid assembly of compound tethers. All novel phosphoramidites described have been successfully used in automated syntheses of 5'-modified oligonucleotides. PMID- 18428960 TI - A convenient stereochemical notation for p-chiral nucleotide analogs. AB - The D(P)/L(P) convention is a stereochemical notation for P-chiral nucleotide analogs and related compounds. In contrast to the absolute R(P)/S(P) notation, the D(P)/L(P) system is based on a geometrical relationship between substituents in a dinucleoside monophosphate skeleton. The D(P)/L(P) notation is a convenient alternative to the R(P)/S(P) notation for stereochemical correlation analysis of physical, chemical, and biological properties of nucleotide and oligonucleotide analogs bearing any type of tri- or tetra-coordinated phosphorus moiety. PMID- 18428961 TI - Synthesis of ribonucleosides by condensation using trimethylsilyl triflate. AB - This unit describes, in detail, the optimized condition for the synthesis of nucleosides making use of the trimethysilyl triflate-mediated silyl-Hilbert Johnson synthesis. This unit focuses on the mechanistic understanding of this universal and conveniently applicable method. PMID- 18428962 TI - Synthesis of 2'-O-beta-d-Ribofuranosylnucleosides. AB - A simple and efficient method for the preparation of 2-O-beta-D ribofuranosylnucleosides, minor tRNA components, is described in this unit. The method consists of condensation of a small excess of 1-O-acetyl-2,3,5-tri-O benzoyl-beta-D-ribofuranose activated with tin tetrachloride with N-protected 3,5 O-tetra-isopropyldisiloxane-1,3-diyl-ribonucleosides in 1,2-dichloroethane. Subsequent deprotection produces 2-O-beta-D-ribofuranosylnucleosides in an overall yield of 46% to 72%. PMID- 18428963 TI - Preparation of 2'-deoxy-2'-methylseleno-modified phosphoramidites and RNA. AB - The derivatization of nucleic acids with selenium is a useful approach to facilitate phase determination during three-dimensional structural analysis by X ray crystallography. This unit describes (1) the synthesis and characterization of 2'-deoxy-2'-methylseleno (2'-Se-methyl) nucleosides and their corresponding 3' O-(2-cyanoethyl)-N,N-diisopropylphosphoramidite derivatives, (2) the site specific incorporation of 2'-Se-methyl ribonucleosides into oligoribonucleotides by chemical RNA solid-phase synthesis, and (3) the enzymatic ligation of Se containing RNA oligonucleotides to produce a biologically relevant RNA sequence. PMID- 18428964 TI - Palladium-catalyzed cross-coupling reactions in c6 modifications of purine nucleosides. AB - This unit describes the cross-coupling methodology for introduction of diverse C substituents to position 6 of purine nucleosides. Protected 6-chloropurine nucleosides undergo Pd-catalyzed cross-coupling reactions with trialkylaluminium, alkylzinc halides, aryl(tributyl)stannanes, and arylboronic acids to give the corresponding 6-substituted intermediates, which can be deprotected by treatment with NaOMe. (Acetyloxy)methylzinc iodide is used for introduction of the hydroxymethyl group, which can further be transformed to fluoromethyl and difluoromethyl groups. Most of the title 6-substituted purine ribonucleosides possess cytostatic and/or anti-HCV activity. PMID- 18428965 TI - Nucleobase-caged phosphoramidites for oligonucleotide synthesis. AB - Caged compounds are biologically active agents bearing a photolabile group in a strategic position, which makes them temporarily inactive. These compounds can then be delivered to a biological sample without immediately generating an effect. When the sample is then irradiated, e.g., with a laser in a (confocal) microscope, the activity of the substance is released with exact spatiotemporal and dose control. This unit deals with the synthesis of protected nucleoside phosphoramidites bearing a caging group on the nucleobase, which prevents the nucleobases from forming normal Watson-Crick base pairs. These amidites can be used to generate caged oligodeoxynucleotides with a transitory local perturbation that adds an element of spatiotemporal control to oligonucleotide-based applications. PMID- 18428966 TI - Synthesis of altritol nucleoside phosphoramidites for oligonucleotide synthesis. AB - This unit describes in detail the optimized preparations of altritol nucleoside phosphoramidite building blocks for oligonucleotide synthesis (aA, aG, aC, aU). D Altritol nucleosides with adenine and uracil bases are obtained by nucleophilic opening of the epoxide ring of 1,5:2,3-dianhydro-4,6-O-benzylidene-D-allitol using the 1,8-diazabicyclo[5.4.0]undec-7-ene salts of the above-mentioned salts, while phase transfer catalysis (18-crown-6, K2CO3) is optimal for alkylation of 2 amino-6-chloropurine. The cytosine nucleoside is synthesized starting from the uracil congener. The 3'-hydroxyl function of hexitol sugar is protected with the benzoyl group. PMID- 18428967 TI - 3,4-diaminobenzophenone matrix for analysis of oligonucleotides by MALDI-TOF mass spectrometry. AB - This unit contains procedures for analysis of oligonucleotides by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry with 3,4 diaminobenzophenone (DABP) as a matrix. This new matrix has demonstrated advantages in the analysis of oligonucleotides. With DABP as a matrix, intact oligonucleotide ions can be readily produced with lower laser powers, resulting in better detection limits, less fragmentation, and fewer alkali-metal ion adducts compared with results obtained using conventional matrices. Minimal fragmentation and fewer alkali-metal ion adducts were seen even at low oligonucleotide concentrations. It was also found that samples prepared with DABP are highly homogenous, therefore reducing the need to find "sweet spots" in MALDI. Finally, excellent shot-to-shot reproducibility, resolution, and signal-to noise ratio are observed using the DABP matrix. PMID- 18428968 TI - RNA secondary structure prediction. AB - Details for predicting secondary structure of RNA sequences using free energy minimization are given. Protocols present the use of the RNAstructure computer program (for PCs) and the mfold server (for Unix platforms). The minimum free energy structure and a set of suboptimal structures with similar free energies are predicted. Prediction of high-affinity oligonucleotide binding sites to a structured RNA target is also presented. PMID- 18428969 TI - Stereoselective synthesis of sugar nucleotides using neighboring group participation. AB - A straightforward, efficient method for the chemical synthesis of sugar nucleotides derived from D-mannose and L-fucose precursors is described. This synthetic strategy involves the coupling of acylated glycosyl bromides with nucleoside 5'-diphosphates, which enables the exploitation of neighboring group participation to exclusively prepare diastereomerically pure sugar nucleotides of desired 1,2-trans anomeric configuration. This is the first stereoselective direct coupling approach to sugar nucleotide synthesis. Following deprotection using triethylamine and purification via C18 reversed-phase ion-pair chromatography, UDP- and GDP-alpha-D-mannose as well as UDP- and GDP-beta-L fucose were obtained in good yield in only four synthetic steps from D-mannose and L-fucose. PMID- 18428970 TI - Synthesis of 2'- and 3'-C-methylribonucleosides. AB - A simple, efficient method for the synthesis of 2'- and 3'-C methylribonucleosides starting from a common precursor is described. This synthesis achieves conversion of 1,2:5,6-di-O-isopropylidene-3-C-methyl-alpha-D allofuranose into 1,2,3-tri-O-acetyl-5-O-benzoyl-3-C-methyl-alpha,beta-D ribofuranose followed by condensation with nucleic acid bases, with a final ammonolysis leading to 3'-C-methylribonucleosides. Alternatively, the same branched allofuranose converted to l,2,3-tri-O-acetyl-5-O-p-methylbenzoyl-2-C methyl-beta-D-ribofuranose, after analogous Vorbruggen condensation and ammonolysis, provides 2'-C-methylribonucleosides. PMID- 18428971 TI - Solid-phase oligonucleotide labeling with DOTA. AB - This unit describes a method to construct oligonucleotide conjugates labeled with 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA) on a solid support. A nucleosidic phosphoramidite that contains a protected DOTA ligand compatible with normal chain assembly is prepared first. As the chain assembly is completed, the oligonucleotide is deprotected and converted to the corresponding gadolinium(III) chelate, resulting in an oligonucleotide conjugate containing the chelate at the 5'-terminus. PMID- 18428972 TI - Synthesis of cidofovir and (S)-HPMPA ether lipid prodrugs. AB - Cidofovir [(S)-1-(3-hydroxy-2-phosphonomethoxypropyl)cytosine] and (S)-HPMPA [(S) 9-(3-hydroxy-2-phosphonomethoxypropyl)adenine] are potent nucleoside phosphonate antiviral agents that are not orally bioavailable unless one or both of their negative charges are masked. This unit describes the synthesis of hexadecyloxypropyl esters of cidofovir and (S)-HPMPA. These prodrugs are readily absorbed after oral administration and are converted intracellularly to the corresponding diphosphates. The hexadecyloxypropyl esters of cidofovir and (S) HPMPA are orally active in animal models of viral infection. Two synthetic strategies are employed. In the first, cyclic cidofovir is coupled to 3 hexadecyloxy-1-propanol using the Mitsunobu reaction (triphenylphosphine, DIAD), followed by basic hydrolysis of the cyclic ester. In the second, the lipid moiety is incorporated into a phosphonate synthon and a stepwise approach is used to assemble the (S)-HPMPA analog. PMID- 18428973 TI - Chemistry of bisSATE mononucleotide prodrugs. AB - On the basis of AZT as a nucleosidic model, the protocols herein describe the synthesis of various bis(S-acyl-2-thioethyl) phosphotriester derivatives. These compounds, bearing transient phosphate-protecting groups, were designed to liberate the corresponding 5'-mononucleotide inside the cell through an esterase mediated activation process. Two synthetic approaches are presented using either phosphoramidite intermediates or esterification of a nucleoside 5'-monophosphate. PMID- 18428974 TI - Universal 2-(4-nitrophenyl)ethyl and 2-(4-nitrophenyl)ethoxycarbonyl protecting groups for nucleosides and nucleotides. AB - A universal blocking group strategy for nucleobases is described, using the 2-(4 nitrophenyl)ethyl (NPE) group for O4-T-, O4-U-, O6-dG-, and O6-G-protection as well as the 2-(4-nitrophenyl)ethoxycarbonyl (NPEOC) group for amino protection in dC, C, dA, A, dG, and G. Conversion into the corresponding 5'-O-dimethoxytrityl derivatives and subsequent phosphitylation to form the fully protected 3'-O-(2 cyanoethyl-N,N-diisopropylphosphoramidites) and 3'-O-(2-(4-nitrophenyl)ethyl-N,N diisopropylphosphoramidites) produces a new class of interesting building blocks for oligonucleotide synthesis. PMID- 18428975 TI - A universal and recyclable solid support for oligonucleotide synthesis. AB - This unit provides a modified phosphoramidite method to synthesize oligodeoxyribonucleotides onto a universal and reusable hydroxyl solid support thanks to the use of deoxyribonucleoside tert-butyl and cyanoethyl phosphoramidites. The nucleoside tert-butyl phosphoramidite allows the introduction of an H-phosphonate diester linkage using the phosphoramidite method. After elongation, the H-phosphonate diester linker is cleaved by transesterification under mild basic conditions to yield an oligonucleotide with free 3'- and 5'-hydroxyls and the starting solid support. Thus, the solid support is easily recycled and used for a subsequent synthesis. In addition, a nucleoside tert-butyl phosphoramidite could be introduced inside the oligonucleotide chain during the elongation to yield a second H-phosphonate diester linkage. After elongation, the two H-phosphonate diester linkages are cleaved, producing two oligonucleotides with free 3'- and 5'-hydroxyls. PMID- 18428976 TI - Stepwise solid-phase synthesis of nucleopeptides. AB - Phosphodiester-linked peptide-oligonucleotide conjugates (nucleopeptides) are obtained by stepwise solid-phase procedures. The peptide is first assembled on a suitably derivatized solid matrix and the oligonucleotide is subsequently elongated at the free hydroxyl group of the linking amino acid. Temporary acid labile and permanent base-labile protecting groups are combined. Careful choice of the protection scheme is required to prevent and minimize side reactions that may degrade the target molecule. PMID- 18428977 TI - Using morpholinos to control gene expression. AB - Morpholino oligonucleotides are stable, uncharged, water-soluble molecules used to block complementary sequences of RNA, preventing processing, read-through, or protein binding at those sites. Morpholinos are typically used to block translation of mRNA and to block splicing of pre-mRNA, though they can block other interactions between biological macromolecules and RNA. Morpholinos are effective, specific, and lack non-antisense effects. They work in any cell that transcribes and translates RNA, but must be delivered into the nuclear/cytosolic compartment to be effective. Morpholinos form stable base pairs with complementary nucleic acid sequences but apparently do not bind to proteins to a significant extent. They are not recognized by any proteins and do not undergo protein-mediated catalysis; nor do they mediate RNA cleavage by RNase H or the RISC complex. This work focuses on techniques and background for using Morpholinos. PMID- 18428978 TI - Synthesis of Peptide-oligonucleotide conjugates by Diels-Alder cycloaddition in water. AB - Peptide-oligonucleotide conjugates incorporating all the nucleobases and trifunctional amino acids are obtained by Diels-Alder reaction between diene modified oligonucleotides (2'-deoxyribo- or ribo-) and malemide-derivatized peptides. Both reagents are easily synthesized by on-column derivatization of the corresponding peptides and oligonucleotides. The cycloaddition reaction is carried out under mild conditions, in aqueous solution at 37 degrees C, affording the desired peptide-oligonucleotide conjugate with high purity and yield. The speed of the reaction depends on the size and composition of both reagents, but it is accelerated by the presence of positively charged amino acids in the peptide fragment. However, a small excess of maleimide-derivatized peptide may be required in some cases to complete the reaction within 8 to 10 hr. PMID- 18428979 TI - Biophysical analysis of triple-helix formation. AB - Two methods for DNA triple-helix analysis are described in this unit: a gel-shift assay based on the slower electrophoretic migration of a triplex in a polyacrylamide gel under nondenaturing conditions, and an optical method in which the thermal denaturation of the triple helix is followed by UV spectrophotometry. Both methods give valuable information on the characteristics of DNA triple-helix formation and triplex stability under different conditions. PMID- 18428980 TI - Preformulation and pharmacokinetic studies on antalarmin: a novel stress inhibitor. AB - The preformulation, solubilization and pharmacokinetic evaluation of antalarmin, a stress inhibitor, have been conducted. Antalarmin has a poor water solubility of less than 1 microg/mL and is weakly basic with an experimentally determined pK(a) of 5.0. Multiple solubilization approaches including pH-control either alone or in combination with cosolvents, surfactants and complexing agents have been investigated. The applicability of lipid-based systems has also been explored. Four formulations, each with a targeted drug loading capacity of 100 mg/mL, show potential for oral administration. Three of these formulations are aqueous solutions (10% ethanol + 40% propylene glycol; 20% cremophor EL; 20% sulfobutylether-beta-cyclodextrin) each buffered at pH 1. The fourth formulation is a lipid-based formulation comprising of 20% oleic acid, 40% cremophor EL and 40% Labrasol. No precipitation was observed following dilution of the four formulations with water and enzyme free simulated gastric fluid. However, only the lipid-based formulation successfully resisted drug precipitation following dilution with enzyme free simulated intestinal fluid. Pharmacokinetic analysis conducted in rats revealed that the 20% cremophor EL solution formulation has a fivefold higher oral bioavailability compared to a suspension formulation. The lipid-based formulation resulted in over 12-fold higher bioavailability as compared to the suspension formulation, the highest amongst the formulations examined. PMID- 18428981 TI - Toughness decrease of PLA-PHBHHx blend films upon surface-confined photopolymerization. AB - The present research investigates the effect of photoinduced grafting reaction on the bulk properties of melt processed poly(L-lactic acid) (PLA)-poly[(3 hydroxybutyrate)-co-(3-hydroxyhexanoate)] (PHBHHx) blend films. PLA-PHBHHx blend films, comprising 10 wt % PHBHHx showed a remarkable toughness improvement. From dynamic mechanical analysis of melt processed PLA-PHBHHx blend films, the blend appears to be noncompatible. Unfortunately, PLA-PHBHHx blend films underwent rapid physical aging as characterized using differential scanning calorimetry, resulting in a significant toughness loss. Physically aged films regained the original toughness on annealing at 60 degrees C for 30 min. Annealed PLA-PHBHHx blend films also underwent physical aging leading to a significant toughness loss. Hydrophilic monomers like acrylic acid and acrylamide were successfully photopolymerized from the film surface using a sequential, two-step photografting approach. The resultant films were characterized using water contact angle goniometry, ATR-FTIR spectroscopy, and mechanical testing. PLA-PHBHHx blend films lost their toughness significantly on surface modification and this was assigned to UV-assisted solvent induced crystallization as characterized using wide-angle X-ray diffraction analyses. PMID- 18428982 TI - Fabrication of drug-loaded electrospun aligned fibrous threads for suture applications. AB - In this work, drug-loaded fibers and threads were successfully fabricated by combining electrospinning with aligned fibers collection. Two different electrospinning processes, that is, blend and coaxial electrospinning, to incorporate a model drug tetracycline hydrochloride (TCH) into poly(L-lactic acid) (PLLA) fibers have been used and compared with each other. The resulting composite ultrafine fibers and threads were characterized through scanning electron microscopy, transmission electron microscopy, Fourier transform infrared spectroscopy, X-ray diffraction, differential scanning calorimetry, and tensile testing. It has been shown that average diameters of the fibers made from the same polymer concentration depended on the processing method. The blend TCH/PLLA fibers showed the smallest fiber diameter, whereas neat PLLA fibers and core shell TCH-PLLA fibers showed a larger proximal average diameter. Higher rotating speed of a wheel collector is helpful for obtaining better-aligned fibers. Both the polymer and the drug in the electrospun fibers have poor crystalline property. In vitro release study indicated that threads made from the core-shell fibers could suppress the initial burst release and provide a sustained drug release useful for the release of growth factor or other therapeutic drugs. On the other hand, the threads from the blend fibers produced a large initial burst release that may be used to prevent bacteria infection. A combination of these results suggests that electrospinning technique provides a novel way to fabricate medical agents-loaded fibrous threads for tissue suturing and tissue regeneration applications. PMID- 18428983 TI - Fabrication and evaluation of a biodegradable proanthocyanidin-crosslinked gelatin conduit in peripheral nerve repair. AB - This study proposed a novel and biodegradable nerve guide conduit in its applicability to peripheral nerve regeneration. A naturally occurring proanthocyanidin (PA) was selected as a cross-linking reagent in preparing the PA crosslinked gelatin (PCG) conduit. Experimental results indicate that 5 wt % of PA was optimal in the complete cross-linking reaction in the PCG conduit. The PCG conduit was brownish and round with a rough outer surface whereas its inner lumen was smooth. The cross-linked networks of the PCG conduit resisted enzymatic hydrolysis under in vitro degradation studies. PA and gelatin were released from the soaked PCG conduit. During the release phase, the concentrations of PA, gelatin, and PCG-soaking solutions were not only nontoxic but also promoted the viability and growth of Schwann cells. The PCG conduit more effectively supported cell attachment and growth. The effectiveness of the PCG conduit as a guidance channel was studied when it was used to repair a 10 mm gap in the rat sciatic nerve. Throughout the 8-week experimental period, the peak amplitude and area under the muscle action potential curve both increased with the recovery period. Histological observations revealed that various regenerated nerve fibers crossed through and beyond the gap region. These results suggest that the PCG conduit can be a candidate for peripheral nerve repair. PMID- 18428984 TI - High-throughput phase-distribution method to determine drug-cyclodextrin binding constants. AB - A high-throughput method has been developed to measure drug-cyclodextrin binding constants. It measures the distribution ratio of a drug between a polymer film [polyvinyl chloride (PVC) with 67% (w/w) dioctyl sebacate (DOS)] and a cyclodextrin-containing buffer in a 96-well format. Measurements of distribution ratios at several cyclodextrin concentrations lead to binding constants. Binding constants for econazole with six CDs have been determined in one 96-well microplate with four replications of each condition in 10 h. The K(1:1)/10(3) M( 1) values are 3.98 +/- 0.13, 3.90 +/- 0.22, 29.3 +/- 2.2, 0.66 +/- 0.04, 1.78 +/- 0.30, 4.08 +/- 0.50, with (2-hydroxyethyl)-beta-cyclodextrin, (2-hydroxypropyl) beta-cyclodextrin, 2,6-di-O-methyl-beta-cyclodextrin, heptakis(2,3,6-tri-O methyl)-beta-cyclodextrin, alpha-cyclodextrin, beta-cyclodextrin, respectively. It is likely that 1:2 complexes are also formed in some cases. This method has also been applied to study the binding behavior as a function of the drug concentration and pH. Binding weakens at higher drug concentration which may be due to the self-association of the drug. An acidic environment decreases the binding constant of CD with the basic econazole. The formation of the 1:2 complexes is completely suppressed in acid as well. This protocol is faster than the phase-solubility method. Moreover, the material requirement is up to four orders of magnitude lower. PMID- 18428985 TI - Absence of the celiac trunk: case report and review of the literature. AB - The authors report a rare variation of the absence of the celiac trunk in a Japanese cadaver, with the left gastric, splenic, common hepatic, and superior mesenteric arteries arising independently from the abdominal aorta in the routine dissection of a 95-year-old Japanese male cadaver. The incidence and developmental and clinical significance of this variation is discussed with a detailed review of the literature. Knowledge of such case has important clinical significance in an abdominal operation or invasive arterial procedure, that is, Appleby procedure and liver transplantation, laparoscopic surgery, and radiological procedures in the upper abdomen. PMID- 18428986 TI - Clubbed fingers: radiological evaluation of the nail bed thickness. AB - It is established that there is an increase in soft tissue of the clubbed digits as demonstrated on previous histopathological examinations. In the present study, the nail bed thickness was assessed and measured on plain radiographs of index fingers in two groups of patients: one group with lung disease and fingers clubbing and one group of normal controls. A vertical x-ray beam was used with a focus-film distance of 1.0 m, with the index finger placed in lateral view directly over the film, without anti-diffusion grid. Three investigators, blinded to prevent bias measured the thickness of soft tissues between the nail root and the terminal phalanx on the radiographs. This method was used to evaluate a group of 85 clinically clubbed (hyponychial angle > 192.0 degrees) adult patients with lung disease and a control group of a 100 normal adult individuals with no clubbing (hyponychial angle < 188.0 degrees). The mean nail bed thickness in the patients with clubbing (n = 85) was 3.88 +/- 0.55 mm (3.00-5.50 mm). In comparison, in the normal subjects (n = 100), the mean was 2.38 +/- 0.27 mm (1.75 3.10 mm), revealing a significant difference (P < 0.001). Only two normal individuals presented nail bed thickness >or=3.0 mm. A good interobserver agreement on the measurements was found (P > 0.900). The radiographic evaluation of the nail bed thickness was easily performed, with good interobserver concordance. It is possible to distinguish between clubbed from nonclubbed fingers, in vivo, using plain radiograph. PMID- 18428987 TI - Demonstration of an anomalous connection between the left coronary artery and the pulmonary artery using a multislice CT 64. AB - In this article, we describe two cases of anomalous connection of the left coronary artery. The first case is an infant of 4 years with an anomalous origin of the left anterior descending (LAD) artery and the diagonal branches and a circumflex artery which originated from the pulmonary artery. The second case is an adult of 50 years with the coronary artery originating from the pulmonary artery. Anomalous origin of the left coronary artery from the pulmonary artery is an uncommon congenital cardiac anomaly with an incidence of 0.25% among all congenital heart defects (Parale and Pawar [2006], J Assoc Physicians India 54:397-399). The originality of this communication consists in the use of a multislice CT scanner as a support for the angiography. The result is an original image with three dimensional details; in the case of the infant, it was determinant in the choice of the surgical approach. PMID- 18428988 TI - Refining the course of the thoracolumbar nerves: a new understanding of the innervation of the anterior abdominal wall. AB - Previous descriptions of the thoracolumbar spinal nerves innervating the anterior abdominal wall have been inconsistent. With modern surgical and anesthetic techniques that involve or may damage these nerves, an improved understanding of the precise course and variability of this anatomy has become increasingly important. The course of the nerves of the anterior abdominal is described based on a thorough cadaveric study and review of the literature. Twenty human cadaveric hemi-abdominal walls were dissected to map the course of the nerves of the anterior abdominal wall. Dissection included a comprehensive tracing of nerves and their branches from their origins in five specimens. The branching pattern and course of all nerves identified were described. All thoracolumbar nerves that innervate the anterior abdominal wall were found to travel as multiple mixed segmental nerves, which branch and communicate widely within the transversus abdominis plane (TAP). This communication may occur at multiple locations, including large branch communications anterolaterally (intercostal plexus), and in plexuses that run with the deep circumflex iliac artery (DCIA) (TAP plexus) and the deep inferior epigastric artery (DIEA) (rectus sheath plexus). Rectus abdominis muscle is innervated by segments T6-L1, with a constant branch from L1. The umbilicus is always innervated by a branch of T10. As such, identification or damage to individual nerves in the TAP or within rectus sheath is unlikely to involve single segmental nerves. An understanding of this anatomy may contribute to explaining clinical outcomes and preventing complications, following TAP blocks for anesthesia and DIEA perforator flaps for breast reconstruction. PMID- 18428989 TI - An innovative approach to supplement the teaching of the spatial gross anatomy relationships of muscles to undergraduates in health sciences. AB - Satisfactory assessment of the musculoskeletal disorders in patients by health professionals, followed by diagnosis, treatment, and referral require a good understanding of spatial and three-dimensional (S&3D) anatomy of musculature. Meanwhile, the importance and impact of S&3D learning has been underscored. A novel method to promote S&3D skills in the learning of the musculature of the upper and lower limb was developed, and subsequently evaluated among 120 first year Physiotherapy and Occupational Therapy undergraduate students at the University of Cape Town. Outlines of human muscles were mapped onto A4 papers, digitalized into a 22-page electronic document, labeled with anatomical details, and printed out. Each student was given a copy to cut out the diagrams and paste them correctly onto adult skeletons using Prestik, in what was called the muscle cut-out exercise (MCOE), just prior to each dissection practical. A questionnaire evaluating the MCOE was developed and 56 students responded. The students "strongly agreed" that the MCOE was useful in teaching them the spatial perspective of the human musculature. Forty-nine percent of the students felt that the size of the diagrams did not always match the skeletons due to variable sizes of the skeletons and 27% suggested the use of color as a means of enhancing the diagrams. The MCOE is a low-cost, enjoyable, time-efficient, easily reproducible, and hands-on practical learning experience. PMID- 18428990 TI - Two peculiar conditions following a coma: a clinical case of heterotopic ossification concomitant with keloid formation. AB - The etiology and formation pattern of heterotopic ossifications (HO) are still unknown. They occur in soft tissues in which bone does not normally form, near one or more proximal joints. In this article, the authors report a peculiar case of a 31-year-old patient affected by scapulo-humeral ankylosis that occurred about 6 months after a coma, in which two unusual concomitant conditions were observed: HO formation in the scapulo-humeral region and the development of keloids during wound repair. The scapulo-humeral ankylosis was resolved surgically with the removal of the HO, which was then studied morphologically to understand its formation pattern. By light microscopy and transmission electron microscopy, it was observed that heterotopic bone displays the normal microscopic structure of primary bone, in which two types of bone tissue were recognized, i.e., woven-fibered bone, deeply located and produced first, and lamellar bone. This suggests that the pattern of HO formation retraces the ontogenetic steps that normally occur during intramembranous ossification. The authors also discuss the peculiar concomitance of HO formation and keloid development, speculating that, although they are different conditions localized in dissimilar regions, they might be hypothetically triggered by a common event, such as the release of factors likely issued during the coma status. PMID- 18428991 TI - Comments on inositol supplementation in pregnancies at risk of apparently folate resistant NTDs. PMID- 18428992 TI - Solid state mechanochemical simultaneous activation of the constituents of the Silybum marianum phytocomplex with crosslinked polymers. AB - Simultaneous improvement of solubilization kinetics of main flavolignans of Silybum marianum extract was obtained cogrinding with two crosslinked polymers: micronized crospovidone, PVP-CL(R) and sodium carboxymethylcellulose, Ac-Di Sol(R) in the 1:3 active-to-polymer weight ratio. By this process it was assessed that the main extract components lost its crystalline structure, and the powder surface area was increased by 2.1- and 1.7-fold in the coground products with Ac Di-Sol(R) and PVP-CL(R), respectively. This activated status of the dry extract remained stable over a period of 2 years. Solubilization kinetics resulted ameliorated both in terms of entire dry extract and in terms of single components. When the 1/3 coground systems with PVP-CL(R) and Ac-Di-Sol(R) were dissolved in saturated conditions they gave a concentration improvement compared to the native product of 8 and 31 times of silybin A, 7 and 27 times of silybin B, whereas in the case of silychristin a double concentration was obtained only using Ac-Di-Sol(R). The in vivo studies on rats confirmed that this solubilization improvement corresponded to an effective oral bioavailability enhancement. The highest bioavailability improvement was obtained with Ac-Di Sol(R), with a relative bioavailability of 88.6, 17.96, and 16.4 compared to the extract for silybin A, silybine B, and silychristine, respectively. PMID- 18428993 TI - Body symmetry and asymmetry in early Greek anatomical reasoning. AB - This historical note focuses on some of the earliest reports of human anatomy found in Greek medical literature. These passages testify the initial steps taken by Greek scientists in building a theoretical model of the human body. In these excerpts, one finds erroneous anatomical descriptions, which shed light on the epistemological approach used by these intellectual pioneers. Because of the lack of systematic dissection, it appears that early Greek anatomists developed a somewhat stylized idea of the human body that used a certain degree of symmetry. Overcoming the concept of a strict left-right bilateral parallelism in human body architecture was a challenging intellectual task that required prolonged observation of dissected corpses. PMID- 18428994 TI - CT study on morphometry of frontal sinus. AB - The aim of this study was to determine the prevalence and morphological characteristics of the frontal sinus in an adult population. This study was conducted retrospectively on paranasal CT scans in the axial and coronal planes of 300 cases (123 male and 177 female). The mean age was 40.74 +/- 13.34 (range 20-83). Measurements of the width, height and anteroposterior length for each sinus and total width were obtained from CT scans. Measurements were compared statistically with relation to side and sex. The cases were divided into subgroups according to age for each sex and each measurement parameter was also compared among the subgroups. All measurements tended to be larger on the left side and were significantly larger in males than females. There was a significant difference in the anteroposterior lengths of right and left sides in both males and females and height for males and width for females. In both sexes, the highest values of measurements were usually observed at the 31-40 age group and there was a tendency to decrease with aging. The larger diameters of the left frontal sinus imply that it may be more possibly violated during surgical interventions. Morphometric features differed significantly in the two sexes at different ages and comparison with previous studies presented great regional variability. The size of the frontal sinus was seen to be related to age and sex. The knowledge provided in the present study is useful for some surgical procedures and widens the anthropometric knowledge of humanity. PMID- 18428995 TI - Morphological study of sinus flow in the confluence of sinuses. AB - The confluence of sinuses (CS; torcular herophili) is represented by the junction of the superior sagittal (SSS), straight (SS), occipital (OS), and two transverse sinuses (TS). The objective of this study was to interpret sinus flow around the CS by morphological investigation of the sinuses. This study is based on visual examination of dural venous sinuses in the region of the CS in 31 adult cadavers. In the inflow zone, we examined the direction of SSS and SS flow. In the communication zone, we examined the extent to which outflow sinuses communicate with other sinuses. In the outflow zone, we used the diameters of outflow sinuses to determine anatomical dominance. The SSS entered the CS via the right TS in 16 cases (51.6%) and via the center of the CS in 14 cases (45.2%). The SS entered via the center of the CS in 18 cases (58.1%) and via the left TS in 11 cases (35.5%). Outflow sinuses communicated freely in 26 cases (83.8%) and communicated partially in five cases (16.2%). Partial communication was the result of a septate CS. In terms of outflow, the right TS was dominant in 11 cases (35.5%), and in 18 cases (58.1%), outflow was symmetrical. The direction of SSS inflow was different from that of SS inflow, and partial communication was observed in five cases (16.1%). Therefore, the presence of a septum may be considered an anatomical factor, with implications in diagnosis or in the sacrifice of the outflow sinus of the CS. PMID- 18428996 TI - Estimation of the number and size of motor units in intrinsic laryngeal muscles using morphometric methods. AB - The number and size of motor units in the intrinsic laryngeal muscles were estimated by morphometric methods. Laryngeal muscles with their respective nerve branches were obtained from 64 fresh cadavers (32 older than 60 years, mean age 74 +/- 9 years and 32 younger than 60 years, mean age 51 +/- 8 years). Myelinated nerve fibers and the total number of muscle fibers were counted. Motor unit size was estimated by dividing the total number of muscle fibers by the total number of motor units in each case. The mean number of motor units ranged from 268 +/- 1.3 (interarytenoid muscle) to 431 +/- 1.6 (cricothyroid muscle). Thyroarytenoid and cricothyroid muscle presented the smallest (9.8 +/- 0.2) and largest (20.5 +/ 0.9) motor unit size, respectively, suggesting that thyroarytenoid muscle has a greater capacity to fine-tune its total force compared with the other intrinsic laryngeal muscles. No differences in motor unit number or size were observed between the right and left sides or between younger and older subjects. It is suggested that synaptic rearrangements may occur at the level of the neuromuscular junction in the human larynx that may explain the age-related changes in motor units reported by clinical methods. PMID- 18428997 TI - Cubital fossa venipuncture sites based on anatomical variations and relationships of cutaneous veins and nerves. AB - Venipuncture is a routine procedure performed at medical institutions for blood collection and blood donation, as well as for health screening and testing. Venipuncture is invasive and usually mildly painful, but it occasionally causes internal bleeding and chronic persistent pain, which is referred to as complex regional pain syndrome. The most common site of venipuncture is the upper arm. The present study macroscopically and anatomically ascertained positional relationships between cutaneous nerves and veins in the cubital (aka antecubital) fossa in many cadaveric dissections to determine the risk of peripheral nerve injury during venipuncture. We identified the most suitable venipuncture site in the upper arm. The medial cutaneous nerve of the forearm (MCNF) passed above the median cubital vein (MCBV) in 27 of 128 cases (21.1%), and was located inferior to the MCBV in 37 of 128 cases (28.9%). The MCBV also passed above the lateral cutaneous nerve forearm (LCNF) in 8 of 128 cases (6.2%). The LCNF was located deeper than the MCBV in 56 of 128 cases (43.8%). The distribution of cutaneous veins and nerves widely varies, and while no single area suitable for all individuals was identified, puncture of the MCBV near the cephalic vein is the least likely to cause nerve damage. PMID- 18428998 TI - Junior doctors' knowledge of applied clinical anatomy. AB - This study examines the level of knowledge of applied clinical anatomy among junior doctors. A multiple-choice questionnaire was designed, which covered 15 areas of anatomical knowledge essential to clinical practice, for example, important surface landmarks and interpretation of radiographs. The questionnaire was completed by 128 individuals. They comprised anatomy demonstrators, preregistration house officers (PRHOs), senior house officers (SHOs) and specialist registrars (SpRs) across the range of medical and surgical specialities. Answers were scored and analyzed by group, allowing comparison not only between newly qualified PRHOs and more senior doctors, but also with anatomy demonstrators who had undergone more traditional anatomical training. The results reveal a wide variation of knowledge among junior doctors, with PRHOs scoring an average of 72.1%, SHOs 77.1%, SpRs 82.4%, and demonstrators 82.9%. This progression in knowledge up the clinical hierarchy may reflect clinical experience building upon the foundations laid in medical school, although with demonstrators topping the league table, it seems that intensive academic training is the most beneficial. With junior doctors' training in the UK currently in flux, these results highlight the need for training in clinical anatomy to hold an important place in the development of tomorrow's clinicians. PMID- 18429000 TI - Disease control intervals in high-risk neuroblastoma. AB - BACKGROUND: Current salvage therapy for recurrent high-risk neuroblastoma is rarely curative. Assessment of the effectiveness of new, primarily cytostatic agents requires the redefinition of study endpoints to reflect disease stabilization rather than tumor response or regression. The intervals of disease control in the patients in the current study with recurrent neuroblastoma were characterized to provide comparison criteria for exploratory studies of new agents. METHODS: Disease control intervals, disease-free survival, postrecurrence survival, and median time to treatment failure were estimated in 90 patients with high-risk neuroblastoma treated between January 1991 and June 2002 on 3 St. Jude neuroblastoma protocols. RESULTS: The estimated median time to disease recurrence was 18.3 months (95% confidence interval [95% CI], 15.9-22.4 months) for the first recurrence, 8.7 months (95% CI, 5.0-12.2 months) for the second recurrence, and 3.8 months (95% CI, 2.5-5.4 months) for the third recurrence. The 5-year estimate of survival after the first disease recurrence was 11%+/-4%. Patients with longer initial disease control had a postrecurrence survival advantage:the 5 year estimated postrecurrence survival was 15.3%+/-6.3% for patients with initial disease control>or=16 months and 8.1%+/-5.5% for others (P=.006). The median disease control interval was approximately halved after each disease recurrence. CONCLUSIONS: The previous disease control interval should be considered in stratification schemes for future phase 2 testing of new agents for the treatment of neuroblastoma. For the optimal evaluation of new treatment strategies that incorporate cytostatic agents, study design and selection of endpoints must take into account the current patterns of recurrence or progression of neuroblastoma. PMID- 18429001 TI - Long-term outcomes in extremity soft tissue sarcoma after a pathologically negative re-resection and without radiotherapy. AB - BACKGROUND: The purpose was to define the rate of local recurrence (LR) and identify prognostic factors for LR in patients with extremity soft-tissue sarcoma (STS) treated with limb-sparing surgery and a pathologically negative re resection specimen without radiotherapy (RT). METHODS: A review of the prospective sarcoma database identified 200 patients with primary, nonmetastatic, extremity STS treated with limb-sparing surgery between June 1982 and December 2002 who had a pathologically confirmed negative re-resection. None of the patients had adjuvant RT. Univariate and multivariate analyses were performed to determine clinicopathologic factors associated with LR. RESULTS: With a median follow-up of 82 months the 5-year actuarial LR rate was 9%. Factors associated with higher LR rates on univariate and multivariate analysis were older age, stage III presentation, and histology. The 5-year LR rate was 5% for those<50 compared with 15% for those>or=50 (P=.001). For patients with stage III the LR rate was 26% versus 7% for those with stage I/II (P<.001). On multivariate analysis only age>or=50 (relative risk [RR] 3.3; P=.02) and stage III disease (RR 3.4; P=.01) remained significant predictors for LR. When the cohort of patients was divided into 3 groups based on the number of risk factors present, the 5-year LRs were as follows: no risk factors (stage I/II and<50 years old) 4%, 1 risk factor (stage III or>or=50) 12%, and 2 risk factors (stage III and>or=50) 31% (P<.01). CONCLUSIONS: Patients with a pathologically negative re-excision represent a heterogeneous group. Whereas the overall rate of local recurrence at 5 years was 9% for the entire cohort, patients with older age and/or stage III disease had a higher rate of LR. Therefore, treatment decisions especially with regard to adjuvant RT should be individualized and not be based solely on the finding of a negative re-resection. PMID- 18429002 TI - Outcomes after radiotherapy for basaloid squamous cell carcinoma of the head and neck: a case-control study. AB - BACKGROUND: Basaloid squamous cell carcinoma (BSCC) is an uncommon, high-grade variant of squamous cell carcinoma (SCC) of the head and neck. Its poorer prognosis compared with common SCC remains controversial. The authors investigated the outcomes of patients with BSCC who received radiotherapy and compared them with the outcomes of patients with SCC. METHODS: From 1994 to 2004, 1007 patients received radiotherapy for head and neck carcinoma with lymph node involvement. The histologic types consisted of 51 BSCC, 431 poorly differentiated SCC (PSCC), and 525 well or moderately differentiated SCC (WMSCC). A case-control analysis was performed with BSCC matched against both PSCC and WMSCC to compare disease-control and survival rates. RESULTS: Patients with BSCC received treatment modalities similar to those received by patients with SCC: They received induction chemotherapy (12%) or concurrent chemotherapy (33%), and a median radiation dose of 70 Gray. Posttreatment viable tumor was present in 44%, 13%, and 28% of neck dissection specimens from patients with BSCC, PSCC, and WMSCC, respectively. The 5-year disease-free survival rates (63%, 77%, and 76%, respectively) and overall survival rates (85%, 70%, and 71%, respectively) demonstrated no statistically significant differences for BSCC, PSCC, or WMSCC, respectively. CONCLUSIONS: In this study, a poorer prognosis could not be demonstrated for irradiated patients with BSCC compared with either PSCC or WMSCC. All patients in this study had positive lymph node status, and the majority of patients (84%) had oropharyngeal cancer. The BSCC cohort did have a relatively high rate of viable tumor in their posttreatment neck dissections, and they had a relatively high rate of distant disease. On the basis of the high rate of lung metastases and the possibility of efficient salvage, the authors recommend obtaining a chest computed tomography scan during initial staging and follow-up. PMID- 18429003 TI - Should patients consider active surveillance? PMID- 18429004 TI - Beta-carotene in multivitamins and the possible risk of lung cancer among smokers versus former smokers: a meta-analysis and evaluation of national brands. AB - BACKGROUND: Some studies have suggested that beta-carotene supplementation may increase the risk of lung cancer, particularly among smokers or former smokers. Beta-carotene, a provitamin A, is available in multivitamins. In the current study, the authors investigated the risk of lung cancer associated with beta carotene in smokers or former smokers and surveyed the beta-carotene content in national brand multivitamins. METHODS: The authors systemically reviewed the published literature using a search of the MEDLINE database and performed a meta analysis of large randomized trials that reported on the effect of beta-carotene supplementation on the incidence of lung cancer among smokers or former smokers. A sample of multivitamins was evaluated for their beta-carotene content and the suggested daily dosage. RESULTS: Four studies contributing 109,394 subjects were available for analysis. The average daily beta-carotene dosage in these trials ranged from 20 to 30 mg daily. Among current smokers, beta-carotene supplementation was found to be significantly associated with an increased risk of lung cancer (odds ratio [OR], 1.24; 95% confidence interval [95% CI], 1.10 1.39). Among former smokers, there was no significant increase noted (OR, 1.10; 95% CI, 0.84-1.45). In a sample of 47 common multivitamins, beta-carotene was present in 70% of the identified formulas. The median dosage of beta-carotene was 0.3 mg (range, 0-17.2 mg) daily. The beta-carotene content was found to be significantly higher among multivitamins sold to improve visual health than among other multivitamins, with a median daily dosage of 3 mg (range, 0-24 mg). CONCLUSIONS: High-dose beta-carotene supplementation appears to increase the risk of lung cancer among current smokers. Although beta-carotene was prevalent in multivitamins, high-dose beta-carotene was observed among multivitamin formulas sold to promote visual health. PMID- 18429005 TI - Reversible features of cervical cancer in human immunodeficiency virus infection: impaired access, impaired surveillance, impaired immunity, impaired outcomes. PMID- 18429006 TI - Peace, Equanimity, and Acceptance in the Cancer Experience (PEACE): validation of a scale to assess acceptance and struggle with terminal illness. AB - BACKGROUND: The role of emotional acceptance of a terminal illness in end-of-life (EOL) care is not known. The authors developed a measure of peaceful acceptance at the EOL, and evaluated the role of peaceful acceptance in EOL decision-making and care. METHODS: The authors developed the Peace, Equanimity, and Acceptance in the Cancer Experience (PEACE) questionnaire to measure the extent to which patients with advanced cancer have a sense of peaceful acceptance of their terminal illness. The scale was administered to 160 patients with advanced cancer along with measures of other attributes that hypothetically are related to acceptance, including cognitive acceptance of terminal illness. EOL outcomes in 56 patients who died during the study also were examined. RESULTS: The 12-item PEACE questionnaire had 2 subscales: a 7-item Struggle With Illness subscale (Cronbach alpha = .81) and a 5-item Peaceful Acceptance subscale (alpha = .78). Both subscales were associated with patients' self-reported peacefulness (correlation coefficient [r] = 0.66 for acceptance [P <.0001]; r = -0.37 for struggle [P < .0001]). Struggle With Illness scores were associated with cognitive terminal illness acknowledgment (mean scores, 14.9 vs 12.4 for patients who were not aware that their illness was terminal; P = .001) and with some aspects of advance care planning (living will or healthcare proxy: mean scores, 13.9 vs 11.5; P = .02). In addition, among patients who had died, the use of a feeding tube at the EOL was associated inversely with Peaceful Acceptance (P = .015). CONCLUSIONS: The current study indicated that the PEACE questionnaire is a valid and reliable measure of peaceful acceptance and struggle with illness. Scores were associated with some choices for EOL care among patients with advanced cancer. PMID- 18429008 TI - Age-related changes in bone-strength-associated geometry indices in naive human population. AB - The aim of the present study was to evaluate age- and sex-related changes in the geometry parameters (metacarpal cortical index (MCI) and Breaking Bending Resistance Index [BBRI]) of long hand bones in a large Chuvashian cohort using cross-sectional and longitudinal study designs. The data were gathered in 1994 (557 individuals) and 2002 (513 individuals). The latter sample included 260 individuals who were studied only during the second expedition, and 253 individuals who were previously investigated in 1994. Statistical analyses included a maximum likelihood-based model-fitting technique and a t-test comparison. Our study describes age-related MCI and BBRI changes in both sexes from the age of 18 years to 84 years. At any age, the BBRI values were higher in males than in females, but MCI was greater in females than in males before age 50 and lower after that age. The study provides initial evidence of a secular trend in MCI and BBRI. In male hand bones, the cortex became relatively thicker and it better resisted bending and breaking in comparison to individuals born at the beginning of the 20th century. In females, the trend toward higher MCI values can be observed only in those born between 1936 and 1966 and the trend toward higher BBRI values stopped in 1950. PMID- 18429009 TI - Magnetization transfer ratio as a predictor of malignancy in breast lesions: preliminary results. AB - MRI is an important tool for investigating breast cancer. Although recognized as the method of choice for screening high-risk patients, and for other indications the role of MRI for lesion characterization remains controversial. Recently some authors have advocated the use of morphologic and postcontrast features for this purpose. Quantitative breast MRI techniques have not been applied extensively in breast diseases. Magnetization transfer (MT) is a quantitative MR technique commonly used to investigate neurological diseases. In breast diseases the use of MT has been limited to improving visualization of areas of enhancement in postcontrast images. The purpose of this study was to evaluate the feasibility and utility of MT in discriminating benign from malignant breast lesions. Fifty two lesions, BIRADS 4 and 5, from 49 patients, were prospectively evaluated using the MT ratio (MTR). Patients were divided into two groups: benign and malignant lesions. The MTR of fat, pectoralis major muscle, fibroglandular tissue, and breast lesions were calculated. A statistically significant difference was found between MTR from benign and malignant lesions (P < 0.001). Preliminary results suggest that MT can be used to evaluate breast lesions. Further studies are necessary to better define the utility and applicability of this technique. PMID- 18429007 TI - Basic evidence of molecular targeted therapy for oral cancer and salivary gland cancer. AB - BACKGROUND: Recently, attention has been focused on molecular targeted cancer therapy in various tumors. Although there is no single consistent molecular target specific for oral squamous cell carcinoma (OSCC) and salivary gland cancer (SGC), there are a number of promising candidate proteins. The aim of this review is to introduce the basic evidences to support the molecular targeting for OSCC and SGC. METHODS: We focused on the 4 molecules, epidermal growth factor receptor (EGFR), cyclooxygenase-2 (COX-2), peroxisome proliferator-activated receptor gamma (PPARgamma), and progesterone receptor, that are, respectively, associated with the proliferation and the differentiation of OSCC and SGC. RESULTS: Gefitinib ("Iressa," ZD1839), a small molecule EGFR tyrosine kinase inhibitor, can inhibit the proliferation of OSCC cell lines in a dose- and time-dependent manner and lead to cell cycle arrest with accumulation of cells in the G1 phase, and a decrease of cells in S phase. The agent suppressed tumor metastasis in the animal model. Furthermore, a cooperative antiproliferative effect was obtained when cancer cells were treated with radiation followed by gefitinib. While radiation alone did not significantly affect p38 mitogen-activated protein kinase and MAP kinase kinase (MEK)1/2 autophosphorylation, the combination of gefitinib and radiation completely inhibited the downstream signaling of EGFR. Gefitinib enhanced tumor radioresponsiveness by multiple mechanisms, including the growth inhibition and effects on DNA repair after exposure to radiation. Next, the level of COX-2 expression correlated inversely with increased tumor radiation sensitivity. Treatment with celecoxib, a COX-2 selective inhibitor, enhanced the radioresponsiveness of HSC-2 cells, which constitutively expressed COX-2. Another promising molecular target is the PPARgamma, which is a member of the nuclear receptor superfamily of ligand-activated transcription factors. Recent studies have demonstrated that PPARgamma ligands induce cellular differentiation and inhibit cell growth in carcinomas of various types. These data suggest that synthetic PPARgamma ligands may be useful for molecular targeting of oral cancer. Finally, the possibility of using molecular targeted therapy directed at hormone receptors in the treatment of advanced SGCs was described. CONCLUSION: The basic data strongly suggested the possibility of tumor suppression by targeting these molecules. Studies of different targeted agents alone or with more conventional treatment modalities are needed to fully determine what role the targeted therapy will play in the management of patients with OSCC and SGC. PMID- 18429010 TI - Evidence for enhanced functional activity of cervical cord in relapsing multiple sclerosis. AB - Functional MRI (fMRI) was used to assess proprioceptive-associated cervical cord activity in 24 relapsing multiple sclerosis (MS) patients and 10 controls. Cord and brain conventional and diffusion tensor (DT) MRI were also acquired. fMRI was performed using a block design during a proprioceptive stimulation consisting of a passive flexion-extension of the right upper limb. Cord lesion number, cross sectional area, mean diffusivity (MD) and fractional anisotropy (FA), whole brain and left corticospinal tract lesion volume (LV), gray matter (GM) MD, and normal appearing white matter (NAWM) MD and FA were calculated. MS patients had higher average cord fMRI signal changes than controls (3.4% vs. 2.7%, P = 0.03). Compared to controls, MS patients also had a higher average signal change in the anterior section of the right cord at C5 (P = 0.005) and left cord at C5-C6 (P = 0.03), whereas no difference was found in the other cord sections. Cord average signal change correlated significantly with cord FA and brain left corticospinal tract LV, GM-MD, and NAWM-FA. This study shows an abnormal pattern of activations in the cervical cord of MS patients following proprioceptive stimulation. Cord fMRI changes might have a role in limiting the clinical consequences of MS associated with irreversible tissue damage. PMID- 18429011 TI - Adaptive 4D MR imaging using navigator-based respiratory signal for MRI-guided therapy. AB - For real-time 3D visualization of respiratory organ motion for MRI-guided therapy, a new adaptive 4D MR imaging method based on navigator echo and multiple gating windows was developed. This method was designed to acquire a time series of volumetric 3D images of a cyclically moving organ, enabling therapy to be guided by synchronizing the 4D image with the actual organ motion in real time. The proposed method was implemented in an open-configuration 0.5T clinical MR scanner. To evaluate the feasibility and determine optimal imaging conditions, studies were conducted with a phantom, volunteers, and a patient. In the phantom study the root mean square (RMS) position error in the 4D image of the cyclically moving phantom was 1.9 mm and the imaging time was approximately 10 min when the 4D image had six frames. In the patient study, 4D images were successfully acquired under clinical conditions and a liver tumor was discriminated in the series of frames. The image quality was affected by the relations among the encoding direction, the slice orientation, and the direction of motion of the target organ. In conclusion, this study has shown that the proposed method is feasible and capable of providing a real-time dynamic 3D atlas for surgical navigation with sufficient accuracy and image quality. PMID- 18429013 TI - Quantitative J-resolved prostate spectroscopy using two-dimensional prior knowledge fitting. AB - Two-dimensional (2D) prior-knowledge fitting (ProFit) was adapted and applied for the quantification of J-resolved (JPRESS) spectra acquired at a field strength of 3T from the human prostate in vivo. In contrast to methods based on simple line fitting and peak integration, commonly applied for metabolite quantification in the prostate, ProFit yields metabolite concentration ratios that are independent of sequence and field strength, since it is based on the linear combination of 2D basis spectra. It is demonstrated that ProFit benefits from the increased information content and reduced baseline distortion in JPRESS prostate spectra, in particular for the quantification of coupled metabolites like citrate (Cit), spermine (Spm), and myo-inositol (mI). The method is validated with 10 repetitive prostate measurements on the same subject. Furthermore, a study carried out on 10 healthy subjects shows that the six prostate metabolites creatine (Cr), total choline (Cho), Cit, Spm, mI, and scyllo-inositol (sI) can be reliably detected in vivo, some of which--especially total Cho and Cit--have proven to be useful markers for the detection of prostate cancer. PMID- 18429012 TI - Seasonal variation and environmental predictors of exhaled nitric oxide in children with asthma. AB - The fraction of exhaled nitric oxide (FeNO), a measure of airway inflammation, shows promise as a noninvasive tool to guide asthma management, but there is a paucity of longitudinal data about seasonal variation and environmental predictors of FeNO in children. The objective of this project was to evaluate how environmental factors affect FeNO concentrations over a 12-month study period among children with doctor diagnosed asthma. We conducted a prospective cohort study of 225 tobacco-smoke exposed children age 6-12 years with doctor-diagnosed asthma including measures of FeNO, medication use, settled indoor allergens (dust mite, cat, dog, and cockroach), and tobacco smoke exposure. Baseline geometric mean FeNO was 12.4 ppb (range 1.9-60.9 ppb). In multivariable analyses, higher baseline FeNO levels, atopy, and fall season were associated with increased FeNO levels, measured 6 and 12 months after study initiation, whereas inhaled steroid use, summer season, and increasing nicotine exposure were associated with lower FeNO levels. In secondary analyses of allergen sensitization, only sensitization to dust mite and cat were associated with increased FeNO levels. Our data demonstrate that FeNO levels over a year long period reflected baseline FeNO levels, allergen sensitization, season, and inhaled steroid use in children with asthma. These results indicate that FeNO levels are responsive to common environmental triggers as well as therapy for asthma in children. Clinicians and researchers may need to consider an individual's baseline FeNO levels to manage children with asthma. PMID- 18429014 TI - Interleaved narrow-band PRESS sequence with adiabatic spatial-spectral refocusing pulses for 1H MRSI at 7T. AB - Proton magnetic resonance spectroscopic imaging ((1)H MRSI) is a useful technique for measuring metabolite levels in vivo, with Choline (Cho), Creatine (Cre), and N-Acetyl-Aspartate (NAA) being the most prominent MRS-detectable brain biochemicals. (1)H MRSI at very high fields, such as 7T, offers the advantages of higher SNR and improved spectral resolution. However, major technical challenges associated with high-field systems, such as increased B(1) and B(0) inhomogeneity as well as chemical shift localization (CSL) error, degrade the performance of conventional (1)H MRSI sequences. To address these problems, we have developed a Position Resolved Spectroscopy (PRESS) sequence with adiabatic spatial-spectral (SPSP) refocusing pulses, to acquire multiple narrow spectral bands in an interleaved fashion. The adiabatic SPSP pulses provide magnetization profiles that are largely invariant over the 40% B(1) variation measured across the brain at 7T. Additionally, there is negligible CSL error since the transmit frequency is separately adjusted for each spectral band. in vivo (1)H MRSI data were obtained from the brain of a normal volunteer using a standard PRESS sequence and the interleaved narrow-band PRESS sequence with adiabatic refocusing pulses. In comparison with conventional PRESS, this new approach generated high-quality spectra from an appreciably larger region of interest and achieved higher overall SNR. PMID- 18429015 TI - 3D RINEPT {1H}-31P CSI: a feasible approach for the study of membrane turnover in the human brain. AB - A fast, full 3D elliptical k-space encoding phosphorous-31 ((31)P) chemical shift imaging sequence, incorporating heteronuclear polarization transfer editing (RINEPT), was established. RINEPT literally requires simultaneous pulses at the frequencies of both the sensitive and the insensitive nuclei, but only a few MR imaging systems feature a second independent transmit channel. In this study, possible signal degradation of a sequential sequence design on systems featuring a single transmit channel was investigated with spin density matrix calculations and phantom measurements. In addition, metabolite signal intensities were determined in vivo as a function of echo and repetition times. The results enable optimization of the signal-to-noise ratio of one or more metabolites of interest. The results convincingly show that the optimized RINEPT sequence is useful in clinical routine (31)PMRS protocols and provides spectra of excellent quality for the study of cell membrane phospholipid turnover in the human brain even at a low field strength of 1.5T. The 3D sequence design allows covering the whole brain in a single measurement, while scan times are compliant with clinical routine protocols. PMID- 18429016 TI - Automatic passive tracking of an endorectal prostate biopsy device using phase only cross-correlation. AB - MR-guided transrectal prostate biopsy is currently a time-consuming procedure because the imaging slice is often manually realigned with the biopsy needle during lesion targeting. In this work a pulse sequence is presented that automatically follows a passive marker attached to a dedicated MR biopsy device holder, thus providing an alternative to existing active tracking methods. In two orthogonal tracking FLASH images of the marker the position of the needle axis is automatically identified using a phase-only cross-correlation (POCC) algorithm. The position information is then used to realign a trueFISP imaging slice in real time. In phantom experiments the sensitivity of this technique to initial misalignments of the marker and to the signal-to-noise ratio was evaluated. In several puncture experiments the precision of the needle placement was analyzed. The POCC algorithm allowed for a precise identification of the marker in the images even under severe initial misalignments of up to 45 degrees. At a frame rate 1 image/s a precision of the needle placement of 1.5 +/- 1.1 mm could be achieved. PMID- 18429018 TI - Generalized k-space decomposition with chemical shift correction for non Cartesian water-fat imaging. AB - Chemical-shift artifacts associated with non-Cartesian imaging are more complex to model and less clinically acceptable than the bulk fat shift that occurs with conventional spin-warp Cartesian imaging. A novel k-space based iterative decomposition of water and fat with echo asymmetry and least-squares estimation (IDEAL) approach is introduced that decomposes multiple species while simultaneously correcting distortion of off-resonant species. The new signal model accounts for the additional phase accumulated by off-resonant spins at each point in the k-space acquisition trajectory. This phase can then be corrected by adjusting the decomposition matrix for each k-space point during the final IDEAL processing step with little increase in reconstruction time. The technique is demonstrated with water-fat decomposition using projection reconstruction (PR)/radial, spiral, and Cartesian spin-warp imaging of phantoms and human subjects, in each case achieving substantial correction of chemical-shift artifacts. Simulations of the point-spread-function (PSF) for off-resonant spins are examined to show the nature of the chemical-shift distortion for each acquisition. Also introduced is an approach to improve the signal model for species which have multiple resonant peaks. Many chemical species, including fat, have multiple resonant peaks, although such species are often approximated as a single peak. The improved multipeak decomposition is demonstrated with water-fat imaging, showing a substantial improvement in water-fat separation. PMID- 18429017 TI - Slice-selective tunable-flip adiabatic low peak-power excitation pulse. AB - Adiabatic pulses are useful in achieving uniform excitation profiles in the presence of B(1)-inhomogeneity. At higher fields, this inhomogeneity becomes more severe, further amplifying the need for B(1)-insensitive excitation. Although gradient modulation techniques for slice-selective adiabatic excitation have been introduced, a pulse that falls within the gradient and RF amplifier limits for most commercial human scanners is currently unavailable. In this work, we present an alternative gradient modulated approach for pulse design that achieves adiabatic slice selection with significantly lower RF peak power requirements. The resulting Slice-selective Tunable-flip AdiaBatic Low peak-power Excitation (STABLE) pulse consists of an oscillating gradient in conjunction with a BIR-4 like RF envelope that is sampled by many short spatial subpulses to achieve spatial selectivity. Simulations show that the expected spatial profile as well as the off-resonance behavior of the pulse remain invariant for a range of B(1) values. Phantom and in vivo results demonstrate the adiabaticity and slice selectivity of the STABLE pulse. PMID- 18429020 TI - Off-resonance-dependent slice profile effects in balanced steady-state free precession imaging. AB - The purpose of this study was the simulation and measurement of balanced steady state free precession (bSSFP) slice profiles for a detailed analysis of the influence of off-resonance effects on slice profile shape and bSSFP signal intensity. Due to the frequency response function of the bSSFP sequence, measurements that are not on-resonance result in broadened effective slice profiles with different off-resonance-dependent shapes and signal intensities. In this study, bSSFP slice profile effects and their dependence on off-resonance were investigated based on bSSFP signal simulations of phantom data as well as blood and tissue. For a better assessment of the similarity of measured and simulated slice profiles the field map was integrated in the slice profile simulations. The results demonstrate that simulations can accurately predict bSSFP slice profiles. Both measurements and simulations indicate that there is a substantial increase in signal intensity close to the banding artifacts, i.e., at spatial locations with off-resonance frequencies corresponding to a dephasing/TR = +/- pi resulting in signal void (bands). For routine bSSFP imaging, off resonance-dependent slice broadening may thus result in a substantial difference between nominal and true slice thickness and lead to spatially varying slice thickness and signal intensities across the imaging slice. PMID- 18429019 TI - High-contrast 3D neonatal brain imaging with combined T1- and T2-weighted MP RAGE. AB - Optimization of magnetization-prepared rapid gradient-echo (MP-RAGE) sequence variations for maximum white matter (WM) versus gray matter (GM) contrast in neonates at 3T was investigated. Numerical simulations were applied to optimize and compare three contrast preparation modules and to assess the effect of phase encoding (PE) order on contrast between WM and thin cortical GM layers. Simulations predict that a new sequence, which combines both T(1)- and T(2) weighting into the contrast preparation and utilizes an interleaved elliptical spiral PE order, should provide the strongest contrast between neonatal WM and cortical GM. This sequence was compared to a conventional MP-RAGE acquisition (i.e., T(1)-weighted preparation, centric PE order) for in vivo imaging of seven preterm newborn infants. Regional measurements of the contrast-to-noise ratio (CNR) between WM and GM demonstrated an increase of 50-70% (depending on GM region) using the new sequence, in good agreement with theoretical predictions. This improved contrast resulted in superior WM versus GM discrimination in intensity-based brain tissue segmentations. PMID- 18429021 TI - Wireless self-gated multiple-mouse cardiac cine MRI. AB - Despite the excellent image-contrast capability of MRI and the ability to synchronize MRI with the murine cardiac cycle, this technique is underused for assessing mouse models of cardiovascular disease because of its perceived cost and complexity. This perception stems, in part, from complications associated with the placement and adjustment of electrocardiographic leads that may interact with gradient pulses and the relatively long acquisition times required with traditional gating schemes. To improve the efficiency and reduce the cost and complexity of using cardiac MRI in mice, we combined wireless self-gating techniques (with which we derived cardiac synchronization signals from acquired data) with an imaging technique that acquires multislice cardiac cine images from four mice simultaneously. As a result, the wireless self-gated acquisitions minimized animal preparation time and improved image quality. The simultaneous acquisition of cardiac cine data from multiple animals greatly increased throughput and reduced costs associated with instrument access. PMID- 18429022 TI - Simultaneous acquisition of MR angiography and venography (MRAV). AB - A dual-echo pulse sequence for simultaneous acquisition of MR angiography and venography (MRAV) is developed. Data acquisition of the second echo for susceptibility-weighted imaging-based MR venography is added to the conventional three-dimensional (3D) time-of-flight (TOF) MRA pulse sequence. Using this dual echo acquisition approach, the venography data can be acquired without increasing the repetition time, and, therefore, the scan duration of routine TOF MRA scans is maintained. The feasibility of simultaneous acquisition of MRAV is presented in brain scans at different spatial resolutions. The effect of spatial resolution on vein-to-background contrast is also demonstrated. Venous contrast is improved in high-resolution (0.52 x 0.52 x 1.6 mm(3)) images compared to that in standard resolution (0.78 x 0.78 x 1.6 mm(3)) images. This MRAV technique enables the acquisition of MR venography without the need of an extra scan or injection of contrast agent in routine clinical brain exams at 3T. PMID- 18429023 TI - High b-value q-space diffusion-weighted MRI of the human cervical spinal cord in vivo: feasibility and application to multiple sclerosis. AB - Q-space analysis is an alternative analysis technique for diffusion-weighted imaging (DWI) data in which the probability density function (PDF) for molecular diffusion is estimated without the need to assume a Gaussian shape. Although used in the human brain, q-space DWI has not yet been applied to study the human spinal cord in vivo. Here we demonstrate the feasibility of performing q-space imaging in the cervical spinal cord of eight healthy volunteers and four patients with multiple sclerosis. The PDF was computed and water displacement and zero displacement probability maps were calculated from the width and height of the PDF, respectively. In the dorsal column white matter, q-space contrasts showed a significant (P < 0.01) increase in the width and a decrease in the height of the PDF in lesions, the result of increased diffusion. These q-space contrasts, which are sensitive to the slow diffusion component, exhibited improved detection of abnormal diffusion compared to perpendicular apparent diffusion constant measurements. The conspicuity of lesions compared favorably with magnetization transfer (MT)-weighted images and quantitative CSF-normalized MT measurements. Thus, q-space DWI can be used to study water diffusion in the human spinal cord in vivo and is well suited to assess white matter damage. PMID- 18429024 TI - Regional metabolite T2 in the healthy rhesus macaque brain at 7T. AB - Although the rhesus macaque brain is an excellent model system for the study of neurological diseases and their responses to treatment, its small size requires much higher spatial resolution, motivating use of ultra-high-field (B(0)) imagers. Their weaker radio-frequency fields, however, dictate longer pulses; hence longer TE localization sequences. Due to the shorter transverse relaxation time (T(2)) at higher B(0)s, these longer TEs subject metabolites to T(2) weighting, that decrease their quantification accuracy. To address this we measured the T(2)s of N-acetylaspartate (NAA), choline (Cho), and creatine (Cr) in several gray matter (GM) and white matter (WM) regions of four healthy rhesus macaques at 7T using three-dimensional (3D) proton MR spectroscopic imaging at (0.4 cm)(3) = 64 mul spatial resolution. The results show that macaque T(2)s are in good agreement with those reported in humans at 7T: 169 +/- 2.3 ms for NAA (mean +/- SEM), 114 +/- 1.9 ms for Cr, and 128 +/- 2.4 ms for Cho, with no significant differences between GM and WM. The T(2) histograms from 320 voxels in each animal for NAA, Cr, and Cho were similar in position and shape, indicating that they are potentially characteristic of "healthy" in this species. PMID- 18429025 TI - Accelerated proton echo planar spectroscopic imaging (PEPSI) using GRAPPA with a 32-channel phased-array coil. AB - Parallel imaging has been demonstrated to reduce the encoding time of MR spectroscopic imaging (MRSI). Here we investigate up to 5-fold acceleration of 2D proton echo planar spectroscopic imaging (PEPSI) at 3T using generalized autocalibrating partial parallel acquisition (GRAPPA) with a 32-channel coil array, 1.5 cm(3) voxel size, TR/TE of 15/2000 ms, and 2.1 Hz spectral resolution. Compared to an 8-channel array, the smaller RF coil elements in this 32-channel array provided a 3.1-fold and 2.8-fold increase in signal-to-noise ratio (SNR) in the peripheral region and the central region, respectively, and more spatial modulated information. Comparison of sensitivity-encoding (SENSE) and GRAPPA reconstruction using an 8-channel array showed that both methods yielded similar quantitative metabolite measures (P > 0.1). Concentration values of N-acetyl aspartate (NAA), total creatine (tCr), choline (Cho), myo-inositol (mI), and the sum of glutamate and glutamine (Glx) for both methods were consistent with previous studies. Using the 32-channel array coil the mean Cramer-Rao lower bounds (CRLB) were less than 8% for NAA, tCr, and Cho and less than 15% for mI and Glx at 2-fold acceleration. At 4-fold acceleration the mean CRLB for NAA, tCr, and Cho was less than 11%. In conclusion, the use of a 32-channel coil array and GRAPPA reconstruction can significantly reduce the measurement time for mapping brain metabolites. PMID- 18429026 TI - Reconstruction of undersampled non-Cartesian data sets using pseudo-Cartesian GRAPPA in conjunction with GROG. AB - Most k-space-based parallel imaging reconstruction techniques, such as Generalized Autocalibrating Partially Parallel Acquisitions (GRAPPA), necessitate the acquisition of regularly sampled Cartesian k-space data to reconstruct a nonaliased image efficiently. However, non-Cartesian sampling schemes offer some inherent advantages to the user due to their better coverage of the center of k space and faster acquisition times. On the other hand, these sampling schemes have the disadvantage that the points acquired generally do not lie on a grid and have complex k-space sampling patterns. Thus, the extension of Cartesian GRAPPA to non-Cartesian sequences is nontrivial. This study introduces a simple, novel method for performing Cartesian GRAPPA reconstructions on undersampled non Cartesian k-space data gridded using GROG (GRAPPA Operator Gridding) to arrive at a nonaliased image. Because the undersampled non-Cartesian data cannot be reconstructed using a single GRAPPA kernel, several Cartesian patterns are selected for the reconstruction. This flexibility in terms of both the appearance and number of patterns allows this pseudo-Cartesian GRAPPA to be used with undersampled data sets acquired with any non-Cartesian trajectory. The successful implementation of the reconstruction algorithm using several different trajectories, including radial, rosette, spiral, one-dimensional non-Cartesian, and zig-zag trajectories, is demonstrated. PMID- 18429027 TI - Fat suppression for 1H MRSI at 7T using spectrally selective adiabatic inversion recovery. AB - Proton magnetic resonance spectroscopic imaging ((1)H MRSI) at 7T offers many advantages, including increased SNR and spectral resolution. However, technical difficulties associated with operating at high fields, such as increased B(1) and B(0) inhomogeneity, severe chemical shift localization error, and converging T(1) values, make the suppression of the broad lipid peaks which can obscure targeted metabolite signals, particularly challenging. Conventional short tau inversion recovery can successfully suppress fat without restricting the selected volume, but only with significant metabolite signal loss. In this work, we have designed two new pulses for frequency-selective inversion recovery that achieve B(1) insensitive fat suppression without degrading the signal from the major metabolites of interest. The first is a spectrally selective adiabatic pulse to be used in a volumetric (1)H MRSI sequence and the second is a spatial-spectral adiabatic pulse geared toward multi-slice (1)H MRSI. Partial interior volume selection may be used in addition to the pulses, to exclude areas with severe B(0) inhomogeneity. Some differences in the spectral profile as well as degree of suppression make each pulse valuable for different applications. 7T phantom and in vivo data show that both pulses significantly suppress fat, while leaving most of the metabolite signal intact. PMID- 18429029 TI - Evaluation of temporal and spatial characteristics of 2D HYPR processing using simulations. AB - Highly constrained back-projection (HYPR) is a data acquisition and reconstruction method that provides very rapid frame update rates and very high spatial resolution for a time series of images while maintaining a good signal-to noise ratio and high image quality. In this study we used simulations to evaluate the temporal and spatial characteristics of images produced using the HYPR algorithm. The simulations demonstrate that spatial accuracy is well maintained in the images and the temporal changes in signal intensity are represented with high fidelity. The waveforms representing signal intensity as a function of time obtained from regions-of-interest placed in simulated objects track the true curves very well, with variations from the truth occurring only when objects with very different temporal behavior are very close to each other. However, even when objects with different temporal characteristics are touching, their influences on each other are small. PMID- 18429028 TI - Resting-state functional connectivity of the rat brain. AB - Regional-specific average time courses of spontaneous fluctuations in blood oxygen level dependent (BOLD) MRI contrast at 9.4T in lightly anesthetized resting rat brain are formed, and correlation coefficients between time course pairs are interpreted as measures of connectivity. A hierarchy of regional pairwise correlation coefficients (RPCCs) is observed, with the highest values found in the thalamus and cortex, both intra- and interhemisphere, and lower values between the cortex and thalamus. Independent sensory networks are distinguished by two methods: data driven, where task activation defines regions of interest (ROI), and hypothesis driven, where regions are defined by the rat histological atlas. Success in these studies is attributed in part to the use of medetomidine hydrochloride (Domitor) for anesthesia. Consistent results in two different rat-brain systems, the sensorimotor and visual, strongly support the hypothesis that resting-state BOLD fluctuations are conserved across mammalian species and can be used to map brain systems. PMID- 18429030 TI - MRI visualization of rodent liver structure and peritoneal adhesion with dialyzate enhancement. AB - This study investigated the use of peritoneal dialysis fluid (dialyzate) as a MR contrast agent to visualize the liver structure and peritoneal adhesion in rats at 7 T. Intraperitoneal injection of dialyzate (approximately 0.1 ml/g) yielded excellent and consistent intraperitoneal enhancement that delineated the liver lobular structure in all rats studied (N = 8). It also allowed the MR detection of peritoneal adhesions that were surgically induced. MR measurements of adhesion surface areas correlated well with the postmortem estimations (R = 0.99; N = 6). Dialyzate persisted in the intraperitoneal cavity for up to 2 days. T(1) and T(2) values of undiluted dialyzate were found to be 3017.5 +/- 35.3 ms and 108.4 +/- 2.0 ms, respectively. These findings demonstrated dialyzate-enhanced MRI as a potentially valuable technique to localize certain activities within liver (such as local tumor metastasis), and to monitor therapeutic interventions (e.g., against peritoneal adhesion) in preclinical research using small animal models. PMID- 18429031 TI - Relaxation-compensated fast multislice amide proton transfer (APT) imaging of acute ischemic stroke. AB - Amide proton transfer (APT) imaging is a variant form of chemical exchange saturation transfer (CEST) imaging that is based on the magnetization exchange between bulk water and labile endogenous amide protons. Given that chemical exchange is pH-dependent, APT imaging has been shown capable of imaging ischemic tissue acidosis, and as such, may serve as a surrogate metabolic imaging marker complementary to perfusion and diffusion MRI. In order for APT imaging to properly diagnose heterogeneous pathologies such as stroke and cancer, fast volumetric APT imaging has to be developed. In this study the evolution of CEST contrast after RF irradiation was solved showing that although the CEST steady state is reached by the apparent longitudinal relaxation rate, the decreases of CEST contrast after irradiation is governed by the intrinsic relaxation constant. A volumetric APT imaging sequence is proposed that acquires multislice images immediately after a single long continuous wave (CW) RF irradiation, wherein the relaxation-induced loss of CEST contrast is compensated for during postprocessing. The proposed technique was verified by numerical simulation, a tissue-like dual-pH phantom, and demonstrated on an embolic stroke animal model. In summary, our study has established a fast volumetric pH-weighted APT imaging technique, allowing further investigation to fully evaluate its diagnostic power. PMID- 18429032 TI - Feasibility of using limited-population-based arterial input function for pharmacokinetic modeling of osteosarcoma dynamic contrast-enhanced MRI data. AB - For clinical dynamic contrast-enhanced (DCE) MRI studies, it is often not possible to obtain reliable arterial input function (AIF) in each measurement. Thus, it is important to find a representative AIF for pharmacokinetic modeling of DCE-MRI data when individual AIF (Ind-AIF) measurements are not available. A total of 16 patients with osteosarcomas in the lower extremity (knee region) underwent multislice DCE-MRI. Reliable Ind-AIFs were obtained in five patients with a contrast injection rate of 2 cc/s and another five patients with a 1 cc/s injection rate. Average AIF (Avg-AIF) for each injection rate was constructed from the corresponding five Ind-AIFs. For each injection rate there are no statistically significant differences between pharmacokinetic parameters of the five patients derived with Ind-AIFs and Avg-AIF. There are no statistically significant changes in pharmacokinetic parameters of the 16 patients when the two Avg-AIFs were applied in kinetic modeling. The results suggest that it is feasible, as well as practical, to use a limited-population-based Avg-AIF for pharmacokinetic modeling of osteosarcoma DCE-MRI data. Further validation with a larger population and multiple regions is desirable. PMID- 18429033 TI - Semiautomatic off-resonance correction in spiral imaging. AB - Spiral scanning is a promising MRI method, but one limitation is that off resonance effects can cause image blurring. Most current off-resonance correction methods for spiral imaging require an accurate field map, which is difficult to obtain in many applications. Automatic methods can perform off-resonance correction without acquiring a field map. However, these methods are computationally inefficient and relatively prone to estimation error. This study describes a new semiautomatic off-resonance correction method that combines an automatic method with a low resolution field map acquisition for off-resonance correction in spiral scanning. Experiments demonstrate that this method is more robust than conventional automatic off-resonance correction and can provide more accurate off-resonance correction than conventional field map based methods. The proposed method is also computationally efficient and has been implemented for online reconstruction. PMID- 18429034 TI - 3D hyperpolarized He-3 MRI of ventilation using a multi-echo projection acquisition. AB - A method is presented for high-resolution 3D imaging of the whole lung using inhaled hyperpolarized (HP) He-3 MR with multiple half-echo radial trajectories that can accelerate imaging through undersampling. A multiple half-echo radial trajectory can be used to reduce the level of artifact for undersampled 3D projection reconstruction (PR) imaging by increasing the amount of data acquired per unit time for HP He-3 lung imaging. The point spread functions (PSFs) for breath-held He-3 MRI using multiple half-echo trajectories were evaluated using simulations to predict the effects of T(2)* and gas diffusion on image quality. Results from PSF simulations were consistent with imaging results in volunteer studies showing improved image quality with increasing number of echoes using up to 8 half-echoes. The 8-half-echo acquisition is shown to accommodate lost breath holds as short as 6 sec using a retrospective reconstruction at reduced resolution and also to allow reduced breath-hold time compared with an equivalent Cartesian trajectory. Furthermore, preliminary results from a 3D dynamic inhalation-exhalation maneuver are demonstrated using the 8-half-echo trajectory. Results demonstrate the first high-resolution 3D PR imaging of ventilation and respiratory dynamics in humans using HP He-3 MR. PMID- 18429036 TI - Multishot diffusion-weighted SPLICE PROPELLER MRI of the abdomen. AB - Multishot FSE (fast spin echo)-based diffusion-weighted (DW)-PROPELLER (periodically rotated overlapping parallel lines with enhanced reconstruction) MRI offers the potential to reduce susceptibility artifacts associated with single-shot DW-EPI (echo-planar imaging) approaches. However, DW-PROPELLER in the abdomen is challenging due to the large field-of-view and respiratory motion during DW preparation. Incoherent signal phase due to motion will violate the Carr-Purcell-Meiboom-Gill (CPMG) conditions, leading to destructive interference between spin echo and stimulated echo signals and consequent signal cancellation. The SPLICE (split-echo acquisition of FSE signals) technique can mitigate non CPMG artifacts in FSE-based sequences. For SPLICE, spin echo and stimulated echo are separated by using imbalanced readout gradients and extended acquisition window. Two signal families each with coherent phase properties are acquired at different intervals within the readout window. Separate reconstruction of these two signal families can avoid destructive phase interference. Phantom studies were performed to validate signal phase properties with different initial magnetization phases. This study evaluated the feasibility of combining SPLICE and PROPELLER for DW imaging of the abdomen. It is demonstrated that DW-SPLICE PROPELLER can effectively mitigate non-CPMG artifacts and improve DW image quality and apparent diffusion coefficient (ADC) map homogeneity. PMID- 18429035 TI - Single-step nonlinear diffusion tensor estimation in the presence of microscopic and macroscopic motion. AB - Patient motion can cause serious artifacts in diffusion tensor imaging (DTI), diminishing the reliability of the estimated diffusion tensor information. Studies in this field have so far been limited mainly to the correction of miniscule physiological motion. In order to correct for gross patient motion it is not sufficient to correct for misregistration between successive shots; the change in the diffusion-encoding direction must also be accounted for. This becomes particularly important for multishot sequences, whereby-in the presence of motion-each shot is encoded with a different diffusion weighting. In this study a general mathematical framework to correct for gross patient motion present in a multishot and multicoil DTI scan is presented. A signal model is presented that includes the effect of rotational and translational motion in the patient frame of reference. This model was used to create a nonlinear least squares formulation, from which the diffusion tensors were obtained using a nonlinear conjugate gradient algorithm. Applications to both phantom simulations and in vivo studies showed that in the case of gross motion the proposed algorithm performs superiorly compared to conventional methods used for tensor estimation. PMID- 18429037 TI - Toward improved grading of malignancy in oligodendrogliomas using metabolomics. AB - In spite of having been the object of considerable attention, the histopathological grading of oligodendrogliomas is still controversial. The determination of reliable biomarkers capable of improving the malignancy grading remains an essential step in working toward better therapeutic management of patients. Therefore the metabolome of 34 human brain biopsies, histopathologically classified as low-grade (LGO, N = 10) and high-grade (HGO, N = 24) oligodendrogliomas, was studied using high-resolution magic angle spinning nuclear magnetic resonance spectroscopy (HRMAS NMR) and multivariate statistical analysis. The classification model obtained afforded a clear distinction between LGOs and HGOs and provided some useful insights into the different metabolic pathways that underlie malignancy grading. The analysis of the most discriminant metabolites in the model revealed the presence of tumoral hypoxia in HGOs. The statistical model was then used to study biopsy samples that were classified as intermediate oligodendrogliomas (N = 6) and glioblastomas (GBMs) (N = 30) by histopathology. The results revealed a gradient of tumoral hypoxia increasing in the following direction: LGOs, intermediate oligodendrogliomas, HGOs, and GBMs. Moreover upon analysis of the clinical evolution of the patients, the metabolic classification seems to provide a closer correlation with the actual patient evolution than the histopathological analysis. PMID- 18429038 TI - Cardiac metabolism measured noninvasively by hyperpolarized 13C MRI. AB - Pyruvate is included in the energy production of the heart muscle and is metabolized into lactate, alanine, and CO(2) in equilibrium with HCO(3) (-). The aim of this study was to evaluate the feasibility of using (13)C hyperpolarization enhanced MRI to monitor pyruvate metabolism in the heart during an ischemic episode. The left circumflex artery of pigs (4 months, male, 29-34 kg) was occluded for 15 or 45 min followed by 2 hr of reperfusion. Pigs were examined by (13)C chemical shift imaging following intravenous injection of 1 (13)C pyruvate. (13)C chemical shift MR imaging was used in order to visualize the local concentrations of the metabolites. After a 15-min occlusion (no infarct) the bicarbonate signal level in the affected area was reduced (25-44%) compared with the normal myocardium. Alanine signal level was normal. After a 45 min occlusion (infarction) the bicarbonate signal was almost absent (0.2-11%) and the alanine signal was reduced (27-51%). Due to image-folding artifacts the data obtained for lactate were inconclusive. These studies demonstrate that cardiac metabolic imaging with hyperpolarized 1-(13)C-pyruvate is feasible. The changes in concentrations of the metabolites within a minute after injection can be detected and metabolic maps constructed. PMID- 18429039 TI - In vitro labeling of glioma cells with gadofluorine M enhances T1 visibility without affecting glioma cell growth or motility. AB - Gadofluorine is a novel macrocyclic, amphiphilic gadolinium-based contrast agent. We found that malignant glioma cells could be labeled in vitro using Gadofluorine without the need for transfection agents or any other additional means. Labeling with Gadofluorine enhanced the visualization of glioma cells in T(1)-weighted sequences, even if the cells had been cultured in medium without Gadofluorine over several days. The intracellular uptake of Gadofluorine was measured and the loss of relevant amounts of Gadofluorine into the cell culture medium was ruled out by MRI. Confocal laser fluorescence microscopy revealed Cy-5-labeled Gadofluorine in the perinuclear cytoplasmic region, but neither within the nucleus nor bound to the cell membrane. Adverse effects of cellular Gadofluorine uptake were ruled out by proliferation and migration assays. Finally, in vivo analyses provided good visibility of labeled glioma cells in T(1)-weighted sequences after intracerebral injection in mice for more than 2 weeks. We thus conclude that Gadofluorine can easily be used to label glioma cells in vitro without affecting glioma cell biology. Gadofluorine provides an interesting alternative for cellular labeling if iron oxide particles are incorporated insufficiently by target cells or if the vicinity of susceptibility artifacts prohibits the use of signal-decreasing contrast agents. PMID- 18429040 TI - System identification theory in pharmacokinetic modeling of dynamic contrast enhanced MRI: influence of contrast injection. AB - Optimization of experimental settings of dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI), like the contrast administration protocol, is of great importance for reliable quantification of the microcirculatory properties, such as the volume transfer-constant K(trans). Using system identification theory and computer simulations, the confounding effects of volume, rate and multiplicity of a contrast injection on the reliability of K(trans) estimation was assessed. A new tracer-distribution model (TDM), based on in vivo data from rectal cancer patients, served to describe the relationship between the contrast agent injection and the blood time-course. A pharmacokinetic model (PKM) was used to describe the relation between the blood and tumor tissue time-courses. By means of TDM and PKM in series, the tissue-transfer function of the PKM was analyzed. As both the TDM and PKM represented low-frequency-pass filters, the energy-density at low frequencies of the blood and tissue time-courses was larger than at high frequencies. The simulations, based on measurements in humans, predict that the K(trans) is most reliable with a high injection volume administered in a single injection, where high rates only modestly improve K(trans). PMID- 18429042 TI - Molecular and functional characterization of eight novel GAA mutations in Italian infants with Pompe disease. AB - We characterized 29 unrelated patients presenting with the severe form of Pompe disease (Glycogen Storage Disease Type II, acid maltase deficiency) and identified 26 pathogenic mutations divided over 28 different genotypes. Among the eight new mutations, five were exonic point mutations (c.572A>G, c.1124G>T, c.1202A>G, c.1564C>G and c.1796C>A) leading to codon changes (p.Y191C, p.R375L, p.Q401R, p.P522A and p.S599Y); two were intronic point mutations (c.-32-3C>A and c.1636+5G>C) affecting mRNA processing; one was a single base deletion (c.742delC) generating a truncated protein (p.L248PfsX20). A comprehensive evaluation, based on different methodological approaches, confirmed the detrimental effect of the eight mutations on the protein and its function. Structural alterations potentially induced by the five missense mutations were also predicted through visual inspection of the atomic model of the GAA protein, in terms of both function and spatial orientation of specific residues as well as disturbance generated by amino acid substitutions. Although the remarkable heterogeneity of the mutational spectrum in Pompe disease was already known, our data demonstrate and confirm the power of molecular and functional analysis in predicting the natural course of Pompe disease. PMID- 18429041 TI - Ventral specification and perturbed boundary formation in the mouse midbrain in the absence of Hedgehog signaling. AB - Although Hedgehog (HH) signaling plays a critical role in patterning the ventral midbrain, its role in early midbrain specification is not known. We examined the midbrains of sonic hedgehog (Shh) and smoothened (Smo) mutant mice where HH signaling is respectively attenuated and eliminated. We show that some ventral (Evx1+) cell fates are specified in the Shh-/- mouse in a Ptc1- and Gli1 independent manner. HH-independent ventral midbrain induction was further confirmed by the presence of a Pax7-negative ventral midbrain territory in both Shh-/- and Smo-/- mice at and before embryonic day (E) 8.5. Midbrain signaling centers are severely disrupted in the Shh-/- mutant. Interestingly, dorsal markers are up-regulated (Wnt1, Gdf7, Pax7), down-regulated (Lfng), or otherwise altered (Zic1) in the Shh-/- midbrain. Together with the increased cell death seen specifically in Shh-/- dorsal midbrains (E8.5-E9), our results suggest specific regulation of dorsal patterning by SHH, rather than a simple deregulation due to its absence. PMID- 18429043 TI - Mutation profile of the CDH23 gene in 56 probands with Usher syndrome type I. AB - Mutations in the human gene encoding cadherin23 (CDH23) cause Usher syndrome type 1D (USH1D) and nonsyndromic hearing loss. Individuals with Usher syndrome type I have profound congenital deafness, vestibular areflexia and usually begin to exhibit signs of RP in early adolescence. In the present study, we carried out the mutation analysis in all 69 exons of the CDH23 gene in 56 Usher type 1 probands already screened for mutations in MYO7A. A total of 18 of 56 subjects (32.1%) were observed to have one or two CDH23 variants that are presumed to be pathologic. Twenty one different pathologic genome variants were observed of which 15 were novel. Out of a total of 112 alleles, 31 (27.7%) were considered pathologic. Based on our results it is estimated that about 20% of patients with Usher syndrome type I have CDH23 mutations. PMID- 18429047 TI - Reduced half-life of holocarboxylase synthetase from patients with severe multiple carboxylase deficiency. AB - Multiple carboxylase deficiency is a clinical condition caused by defects in the enzymes involved in biotin metabolism, holocarboxylase synthetase (HLCS) or biotinidase. HLCS deficiency is a potentially fatal condition if left untreated, although the majority of patients respond to oral supplementation of 10-20 mg/day of biotin. Patients who display incomplete responsiveness to this therapy have a poor long-term prognosis. Here we investigated cell lines from two such HLCS deficient patients homozygous for the c.647T>G p.L216R allele. Growth of the patients' fibroblasts was compromised compared with normal fibroblasts. Also the patient cells were not sensitive to biotin-depletion from the media, and growth rates could not be restored by re-administration of biotin. The molecular basis for the HLCS deficiency was further investigated by characterisation of the p.L216R protein. The HLCS mRNA was detected in MCD and normal cell lines. However, protein and enzyme activity could not be detected in the patients' cells. In vitro kinetic analysis revealed that enzyme activity was severely compromised for recombinantly expressed p.L216R and could not be increased by additional biotin. Furthermore, the turn-over rate for the mutant protein was double that of wildtype HLCS. These results help provide a molecular explanation for the incomplete biotin-responsiveness of this p.L216R form of HLCS. PMID- 18429048 TI - Haplotype analysis suggests a single Balkan origin for the Gaucher disease [D409H;H255Q] double mutant allele. AB - Gaucher disease is an autosomal recessive lysosomal storage disease that is mainly due to mutations in the GBA gene. Most of the mutant alleles described so far bear a single mutation. However, there are a few alleles bearing two or more DNA changes. It has been reported that patients homozygous for the [D409H;H255Q] double mutant allele (HGVS-approved nomenclature, p.[D448H;H294Q]) present a more severe phenotype than patients homozygous for the relatively common D409H mutation. In this study, we confirmed the detrimental cumulative effect of these two mutations at the enzymatic activity level by the heterologous expression of the single and double mutant alleles. Additionally, we found a high frequency of the [D409H;H255Q] allele in patients from the Balkans and the Adriatic area of Italy. This prompted us to perform a haplotype analysis, using five microsatellite polymorphisms close to the GBA gene, to determine the origin of this allele. The results of the 37 chromosomes analysed showed that most of them share a common haplotype and are consistent with a single origin in the Balkans and the Adriatic area of Italy for the [D409H;H255Q] allele. PMID- 18429049 TI - Recurrent transfusion-related acute lung injury after fresh frozen plasma in a patient with hereditary factor V deficiency. PMID- 18429046 TI - Molecular and developmental biology of the hemangioblast. AB - The hemangioblast hypothesis was proposed a century ago. The existence of hemangioblasts is now demonstrated in mouse and human embryonic stem cell (ESC) derived embryoid bodies (EBs), in the mouse and zebrafish gastrula, and in adults. The hemangioblast is believed to derive from mesodermal cells, and is enriched in the Bry+Flk1+ and Flk1+Scl+ cell populations in EBs and in the posterior primitive streak of the mouse gastrula and in the ventral mesoderm of the zebrafish gastrula. However, recent studies suggest that the hemangioblast does not give rise to all endothelial and hematopoietic lineages in mouse and zebrafish embryos. Although several signaling pathways are known to involve the generation of hemangioblasts, it remains largely unknown how the hemangioblast is formed and what are the master genes controlling hemangioblast development. This review will summarize our current knowledge, challenges, and future directions on molecular and developmental aspects of the hemangioblast. PMID- 18429051 TI - Essential thrombocythemia, polycythemia vera, and myelofibrosis: current management and the prospect of targeted therapy. AB - The recent discovery of JAK2 and/or MPL mutations in polycythemia vera (PV), essential thrombocythemia (ET), and primary myelofibrosis (PMF) has had a major impact on how we diagnose and treat these disorders. For instance, the presence of a JAK2 mutation is now considered conditio sine qua non for the diagnosis of PV and the World Health Organization classification system has recently revised its diagnostic criteria for PV, ET, and PMF to include JAK2 and MPL mutations as clonal markers. From the standpoint of treatment, JAK-STAT is now identified as a legitimate target pathway for drug development in myeloproliferative neoplasms. Herein, I will first outline my views regarding current management in ET, PV, and PMF and then discuss emerging data on preclinical and clinical activity of anti JAK2 small molecule drugs. Am. J. Hematol., 2008. (c) 2008 Wiley-Liss, Inc. PMID- 18429050 TI - Serum ferritin concentrations and body iron stores in a multicenter, multiethnic primary-care population. AB - How often elevated serum ferritin in primary-care patients reflects increased iron stores (normally 0.8 g in men, 0.4 g in women) is not known. The Hereditary Hemochromatosis and Iron Overload Screening (HEIRS) study screened 101,168 primary-care participants (44% Caucasians, 27% African-Americans, 14% Asians/Pacific Islanders, 13% Hispanics, 2% others). Follow-up clinical evaluation was performed in 302 of 333 HFE C282Y homozygotes regardless of iron measures and 1,375 of 1,920 nonhomozygotes with serum ferritin >300 microg/L (men), >200 microg/L (women) and transferrin saturation >50% (men), >45% (women). Quantitative phlebotomy was conducted in 122 of 175 C282Y homozygotes and 122 of 1,102 nonhomozygotes with non-transfusional serum ferritin elevation at evaluation. The estimated prevalence in the Caucasian population of C282Y homozygotes with serum ferritin >900 microg/L at evaluation was 20 per 10,000 men and 4 per 10,000 women; this constellation was predictive of iron stores >4 g in men and >2 g in women. The estimated prevalence per 10,000 of non-C282Y homozygotes with serum ferritin >900 microg/L at evaluation was 7 among Caucasians, 13 among Hispanics, 20 among African Americans, and 38 among Asians and Pacific Islanders, and this constellation was predictive of iron stores >2 g but <4 g. In conclusion, serum ferritin >900 microg/L after initial elevations of both serum ferritin and transferrin saturation is predictive of mildly increased iron stores in multiple ethnic populations regardless of HFE genotype. Serum ferritin >900 microg/L in male C282Y homozygotes is predictive of moderately increased iron stores. PMID- 18429052 TI - Jordans' anomaly in a new neutral lipid storage disease. PMID- 18429053 TI - Inflammation resolution: does the bone marrow have a say? PMID- 18429054 TI - Autologous stem cell transplantation in patients of 70 years and older with multiple myeloma: Results from a matched pair analysis. AB - High-dose therapy and autologous stem cell transplant (HDT) have been shown to prolong survival in multiple myeloma (MM) in randomized trials, but only included patients of 65 years or younger. Given the median age at diagnosis of 66 years, it is important to have a better understanding of the outcome of transplantation in the older patients. We identified 33 patients with MM, who were > or =70 years at the time of their HDT. We matched them to a group of 60 patients, 65 years or younger, (two controls for each patient), based on time to transplant, disease status at transplant, Durie-Salmon stage, labeling index, presence of cytogenetic abnormalities, and presence of circulating plasma cells. The median age of the two groups were 55.6 (range, 37.3-64.9) and 71.7 (range 70-75.8) years at transplant. Although more of the older patients received dose reduced melphalan, the overall response rate was similar (97% vs. 98%) as was the median time to progression (28.5 months vs. 17.8 months, P = 0.7) for the elderly group compared to the younger patients. The median overall survival from transplant was not reached for the elderly patient group compared to 53.2 months for the younger patients, P = 0.7. HDT is feasible in selected patients with multiple myeloma over 70 years. The toxicity of transplant as well as the outcome appears comparable to younger patients. Patients with MM should not be excluded from HDT solely on the basis of their chronological age. PMID- 18429055 TI - Identification of inflammatory and proresolving lipid mediators in bone marrow and their lipidomic profiles with ovariectomy and omega-3 intake. AB - Newly described lipoxygenase (LOX)-generated lipid mediators, that is, resolvins and protectins as well as lipoxins, are both anti-inflammatory and proresolving. We aimed to determine whether these lipid mediators are present in bone marrow and whether their lipidomic profiles are altered following ovariectomy or dietary supplementation with eicosapentaenoic acid (EPA) or docosahexaenoic acid (DHA) ethyl esters. Female rats were ovariectomised or sham-operated. Shams and one ovariectomised group received a diet devoid of omega-3 long-chain polyunsaturated fatty acids. The remaining ovariectomised rats received either 0.5 g EPA or DHA ethyl ester/kg body weight/day for 4 months. Bone marrow was analyzed using both GC to determine fatty acid composition and mediator lipidomics by LC/MS/MS profiling for the presence of LOX-pathway lipid mediators derived from arachidonic acid (AA), EPA, and DHA. LOX-derived products including lipoxins, resolvin D1, resolvin E1, and protectin D1 were identified in bone marrow by the presence of diagnostic ions in their corresponding MS-MS spectra. The proportion of AA relative to DHA and of AA-derived relative to DHA-derived mediators in bone marrow was higher in ovariectomised compared to sham-operated rats. DHA or EPA ethyl ester supplementation increased the percentage of DHA and EPA in bone marrow and increased the proportion of LOX mediators biosynthesized from DHA or EPA, respectively. Given the potent bioactivities of the lipoxins, resolvins, and protectins, the presence and changes in profile postovariectomy and with EPA and DHA ethyl ester supplementation may be of interest in bone marrow function and as a potential source of these mediators in vivo. PMID- 18429056 TI - Analysis of histone modifications by mass spectrometry. AB - Histone N-termini undergo diverse post-translational modifications that significantly extend the information potential of the genetic code. Moreover, they appear to mark specific chromatin regions, modulating epigenetic control, lineage commitment, and overall function of chromosomes. It is widely accepted that histone modifications affect chromatin function, but the exact mechanisms of how modifications on histone tails and specific combinations of modifications are generated, and how they cross-talk with one another, is still enigmatic. Mass spectrometry is ideal for the analysis of histone modifications and is becoming the gold standard for histone post-translational modification analysis since it allows the quantification of modifications and combinations. This unit describes how high-resolution mass spectrometry can be used to study histone post translational modifications. PMID- 18429057 TI - Detection and analysis of protein-protein interactions of organellar and prokaryotic proteomes by blue native and colorless native gel electrophoresis. AB - Native gels enable the analysis of protein complexes on a proteome-wide scale in a single experiment. The protocols described in this unit are based on separation of protein complexes by blue native polyacrylamide electrophoresis (BN-PAGE), the most versatile native gel system, and the closely related milder colorless native PAGE (CN-PAGE). Both BN-PAGE and CN-PAGE are described on analytical to preparative scales. In addition, methods for subsequent analysis of protein complexes are given, including electroelution from native gels as well as denaturing and native two-dimensional PAGE. Finally, the removal of Coomassie dye from electroeluted proteins is detailed along with a discussion of fundamental considerations for the solubilization of membrane protein complexes from various biological samples, which are exemplified for mitochondria, chloroplasts (thylakoids), and cyanobacteria. PMID- 18429058 TI - Utilizing peptide SPOT arrays to identify protein interactions. AB - SPOT arrays consist of synthesized peptides 12- to 18-amino acids long, with overlapping sequences that cover the entire sequence of a protein, covalently linked to a solid support. This unit describes how to construct peptide SPOT arrays, biotinylate recombinant proteins, and conduct overlay assays to identify binding interactions. In addition, directions describing how to analyze results to determine single amino acid binding contributions are included. The two techniques in this unit describe how to scan protein sequences to find binding motifs and how to conduct site-directed mutagenesis studies. PMID- 18429059 TI - Phage display for generating peptide reagents. AB - This unit presents detailed protocols for selection and propagation of landscape phages, which are fusions of filamentous phage fd (or its close relatives M13 and f1) and foreign DNA that result in chimeric phage virions with foreign peptides (8 to 9 amino acids long) covering the entire surface of the phage particles. These landscape phages bind specifically to mammalian and bacterial cells, spores, or discrete molecular targets. PMID- 18429060 TI - MultiBac: multigene baculovirus-based eukaryotic protein complex production. AB - Multiprotein complexes are an emerging focus in current biology, resulting in a demand for advanced heterologous expression systems. This unit provides protocols for the expression of eukaryotic multiprotein complexes using multigene expression vectors. Homologous and site-specific recombinases facilitate their assembly. Thus, modification of individual subunits for revised expression studies is achieved with comparative ease. The strategy outlined here employs the MultiBac baculoviral expression system for multiprotein complexes as an example. Baculoviral expression does not require particular safety precautions due to the replication incompetence of baculovirus in mammalian hosts. The MultiBac system provides for improved protein production due to deletion of specific viral genes (V-cath, chiA). Most of the steps described in this unit are tailored for high throughput approaches. The general strategy of rapidly combining encoding DNAs by recombination into multigene expression vectors for protein complex expression can also be applied to other prokaryotic or mammalian expression systems. PMID- 18429061 TI - Characteristics of amino acids. AB - The physical properties of the amino acids determine the structure and function of the proteins in which they are found. This appendix presents some useful details and relevant physical characteristics of the natural amino acids and modified amino acids (i.e., chemical structures, explanations of the role these structures play in enzymes, relative hydrophobicities, shapes and volumes, and values for some common mass changes due to post-translational modifications). PMID- 18429062 TI - Commonly used detergents. AB - This appendix lists some common detergents along with useful information regarding the type of hydrophilic group (anionic, cationic, amphoteric, or nonionic), the critical micelle concentration (CMC) and the micelle molecular weight. PMID- 18429063 TI - Conversion factors and half-life information for radioactivity. AB - Discussion of radioactivity may use different terms for measurements of radioactivity and dose. Conversion factors for these terms are presented in this appendix. In addition, when investigators are working with a particular radiolabeled compound, it is necessary to calculate the amount of remaining radioactivity based on the amount of time that has elapsed from the reference date and the half life of the radioisotope. Decay factors for the commonly used shorter half life radioisotopes are provided here. PMID- 18429064 TI - Common conversion factors. AB - This appendix lists some of the more common conversion factors for units of measure used throughout Current Protocols manuals, and gives prefixes indicating powers of ten for SI units. PMID- 18429065 TI - Safe use of radioisotopes. AB - In all cases where radioisotopes are used, depending on the quantity and nature of the isotope, certain precautions must be taken to ensure the safety of the scientist. This appendix outlines a few such considerations relevant to the isotopes most frequently used in biological research. PMID- 18429066 TI - Centrifuges and rotors. AB - This appendix provides r(max) values (the maximum value of the rotating radius between the particle being centrifuged and the axis of rotation) for commonly used rotors manufactured by Du Pont (Sorvall), Beckman, Fisher, and IEC. Additionally, nomograms are provided for determining the relative centrifugal force (RCF) where speed and r(max) are known, or the speed where RCF and r(max) are known. PMID- 18429067 TI - Standard laboratory equipment. AB - This appendix lists standard pieces of equipment in the modern biochemistry laboratory, i.e., items used extensively in this manual and thus not included in the individual materials lists. PMID- 18429068 TI - Commonly used reagents. AB - This collection describes the preparation of buffers and reagents used in the manipulation of proteins. PMID- 18429069 TI - Use of protein folding reagents. AB - The reagents and methods for purification of the denaturants guanidine hydrochloride (guanidine-HCl) and urea are described. Sulfhydryl reagents (reducing agents) and "oxido-shuffling" (or oxidative regeneration) systems are also discussed. PMID- 18429070 TI - Dialysis. AB - This appendix describes dialysis of large- and small-volume samples using cellulose membranes with pore sizes designed to exclude molecules above a selected molecular weight. A support protocol describes preparation of membranes for dialysis. PMID- 18429071 TI - Importing biological materials. AB - This overview discusses critical issues regarding importing of restricted biological materials along with criteria for handling these materials. Guidelines for importing non-restricted biological materials are also covered. Recommendations are given for packaging biological materials for export, and finally, the necessary steps for obtaining an import permit application are outlined. PMID- 18429072 TI - Silanizing glassware. AB - Untreated glass contains silicate and silanol groups that can act as ion-exchange and nucleophilic centers. To mask these groups and decrease the hydrophilicity of the surface, various reactive silanes are frequently used to coat the glass surface. This appendix describes a procedure for silanizing glassware. PMID- 18429073 TI - Protein precipitation using ammonium sulfate. AB - The basic theory of protein precipitation by addition of ammonium sulfate is presented and the most common applications are listed. Numerous tables are provided for calculating the appropriate amount of ammonium sulfate to add to a particular protein solution. PMID- 18429074 TI - Statistics: detecting differences among groups. AB - Some of the statistical tests most commonly used (and misused) in biological research are presented here. The tests discussed are those used for comparisons among groups (e.g., t test and ANOVA). The purpose of this appendix is to enable the investigator to determine rapidly the most appropriate way to analyze data, and to point out some of the most common errors to avoid. PMID- 18429075 TI - Analyzing radioligand binding data. AB - A radioligand is a radioactively labeled drug that can associate with a receptor, transporter, enzyme, or any protein of interest. Measuring the rate and extent of binding provides information on the number of binding sites, and their affinity and accessibility for various drugs. Radioligand binding experiments are easy to perform, and provide useful data in many fields. For example, radioligand binding studies are used to study receptor regulation, investigate receptor localization in different organs or regions using autoradiography, categorize receptor subtypes, and probe mechanisms of receptor signaling. This unit reviews the theory of receptor binding and explains how to analyze experimental data. Since binding data are usually best analyzed using nonlinear regression, this unit also explains the principles of curve fitting with nonlinear regression. PMID- 18429076 TI - Media preparation and bacteriological tools. AB - This appendix presents recipes for basic bacteriological media. PMID- 18429077 TI - Growth in liquid or solid media. AB - This appendix presents basic procedures for growing overnight (and larger) cultures, monitoring growth, and titering and isolating bacterial cultures by serial dilution. In addition, protocols are provided for isolating single colonies by streaking and spreading a plate. Replica plating and strain storage/retrieval is also detailed. PMID- 18429078 TI - Preparation of plasmid DNA. AB - This appendix presents basic procedures for alkaline lysis minipreps, both in tubes and in microtiter plates, and also provides an Alternate protocol for boiling lysis. A Support Protocol describes storage of plasmid DNA. PMID- 18429079 TI - Introduction of plasmid DNA into cells. AB - This appendix presents a procedure for transformation using calcium chloride, and also an alternate procedure for one-step preparation and transformation of competent cells. PMID- 18429080 TI - Purification and concentration of DNA from aqueous solutions. AB - This unit presents basic procedures for manipulating solutions of single- or double-stranded DNA through purification and concentration steps. Phenol extraction with ethanol precipitation is described along with an alternate procedure of precipitating DNA with isopropanol. A Support Protocol describes concentration of DNA using butanol. PMID- 18429081 TI - Agarose gel electrophoresis. AB - This appendix presents a protocol for separating and purifying DNA fragments between 0.5 and 25 kb. A Support Protocol describes the use of midigels and minigels. PMID- 18429082 TI - Southern blotting. AB - Southern blotting is the transfer of DNA fragments from an electrophoresis gel to a membrane support (the properties and advantages of the different types of membrane, transfer buffer, and transfer method are discussed in detail), resulting in immobilization of the DNA fragments, so the membrane carries a semipermanent reproduction of the banding pattern of the gel. After immobilization, the DNA can be subjected to hybridization analysis, enabling bands with sequence similarity to a labeled probe to be identified. This appendix describes Southern blotting via upward capillary transfer of DNA from an agarose gel onto a nylon or nitrocellulose membrane, using a high-salt transfer buffer to promote binding of DNA to the membrane. With the high-salt buffer, the DNA becomes bound to the membrane during transfer but not permanently immobilized. Immobilization is achieved by UV irradiation (for nylon) or baking (for nitrocellulose). A Support Protocol describes how to calibrate a UV transilluminator for optimal UV irradiation of a nylon membrane. An alternate protocol details transfer using nylon membranes and an alkaline buffer, and is primarily used with positively charged nylon membranes. The advantage of this combination is that no post-transfer immobilization step is required, as the positively charged membrane binds DNA irreversibly under alkaline transfer conditions. The method can also be used with neutral nylon membranes but less DNA will be retained. A second alternate protocol describes a transfer method based on a different transfer-stack setup. The traditional method of upward capillary transfer of DNA from gel to membrane described in the first basic and alternate protocols has certain disadvantages, notably the fact that the gel can become crushed by the weighted filter papers and paper towels that are laid on top of it. This slows down the blotting process and may reduce the amount of DNA that can be transferred. The downward capillary method described in the second alternate protocol is therefore more rapid than the basic protocol and can result in more complete transfer. Although the ease and reliability of capillary transfer methods makes this far and away the most popular system for Southern blotting with agarose gels, it unfortunately does not work with polyacrylamide gels, whose smaller pore size impedes the transverse movement of the DNA molecules. The third alternate protocol describes an electroblotting procedure that is currently the most reliable method for transfer of DNA from a polyacrylamide gel. Dot and slot blotting are also described. PMID- 18429083 TI - Hybridization analysis of DNA blots. AB - Restriction endonucleases recognize short DNA sequences and cleave double stranded DNA at specific sites within or adjacent to the recognition sequences. Restriction endonuclease cleavage of DNA into discrete fragments is one of the most basic procedures in molecular biology. This appendix describes restriction endonucleases and their properties. PMID- 18429084 TI - Digestion of DNA with restriction endonucleases. AB - Restriction endonucleases recognize short DNA sequences and cleave double stranded DNA at specific sites within or adjacent to the recognition sequences. Restriction endonuclease cleavage of DNA into discrete fragments is one of the most basic procedures in molecular biology. This appendix describes restriction endonucleases and their properties. PMID- 18429085 TI - Quantitation of nucleic acids with absorption spectroscopy. AB - This appendix describes a protocol for measuring absorption of nucleic acid samples at several different wavelengths to assess purity and concentration. A(260) measurements are quantitative for relatively pure nucleic acid preparations in microgram quantities. Absorbance readings cannot discriminate between DNA and RNA; however, the ratio of A at 260 and 280 nm can be used as an indicator of nucleic acid purity. PMID- 18429086 TI - Growth and manipulation of yeast. AB - This unit describes preparation of selected media for growing yeast and also discusses strain storage and revival. Protocols are provided for the assay of beta-galactosidase in liquid culture and for transformation using lithium acetate. PMID- 18429087 TI - Theoretical aspects of the quantitative characterization of ligand binding. AB - Living organisms grow, differentiate, reproduce, and respond to their environment via specific and integrated interactions between biomolecules. The investigation of molecular interactions therefore constitutes a major area of biochemical study, occupying a ubiquitous and central position between molecular physiology on the one hand and structural chemistry on the other. While specificity resides in the details of structural recognition, the dynamic interplay between biomolecules is orchestrated precisely by the thermodynamics of the biomolecular equilibria involved. A common set of physicochemical principles applies to all such phenomena, irrespective of whether the interaction of interest involves an enzyme and its substrate or inhibitor, a hormone or growth factor and its receptor, an antibody and its antigen, or, indeed, the binding of effector molecules that modulate these interactions. The binding affinity, binding specificity, number of binding sites per molecule, as well as the enthalpic and entropic contributions to the binding energy are common parameters that assist an understanding of the biochemical outcome. This unit aims to provide an overview of the design and interpretation of binding experiments. PMID- 18429088 TI - Overview of protein purification and characterization. AB - This insightful overview by one of the most respected names in protein research discusses a broad array of the aspects involved with protein purification, including historical background, determining the purpose for purifying a particular protein, and actual methods and recommendations for purification procedures. Also covered are methods of analysis and detection, and techniques for characterizing a purified protein. Finally, a useful discussion of the equipment needed in a protein purification lab is provided. PMID- 18429089 TI - Strategies for protein purification. AB - This discussion unit presents information on classification of proteins according to shape, oligomeric structure, and function. In addition, strategies for purifying different types of proteins are discussed based on their location and associations within the cell. Specifically, critical issues are detailed for extracellular, cytoplasmic, and membrane-associated proteins, as well as for insoluble proteins. Finally, strategies for the purification of both soluble and insoluble (inclusion bodies) recombinantly expressed proteins are provided. PMID- 18429090 TI - Protein purification flow charts. AB - Protein purification flow charts are presented to give a broad outline of the methods used for different types of proteins. They cannot give any detail, as the process appropriate for each protein will have its own variations at each stage. In most cases, the first stage is to obtain a solution containing the desired protein, after which it can be fractionated by various separation techniques. This unit provides charts tailored for purification of soluble and insoluble recombinant and non-recombinant proteins, as well as membrane-associated proteins. PMID- 18429091 TI - One-dimensional SDS gel electrophoresis of proteins. AB - Electrophoresis is used to separate complex mixtures of proteins, to investigate subunit compositions, and to verify homogeneity of protein samples. It can also serve to purify proteins for use in further applications. In polyacrylamide gel electrophoresis, proteins migrate in response to an electrical field through pores in the gel matrix; pore size decreases with higher acrylamide concentrations. The combination of gel pore size and protein charge, size, and shape determines the migration rate of the protein. This unit contains protocols that gives the standard Laemmli method for discontinuous gel electrophoresis under denaturing conditions, and the standard method for full-size gels is adapted for the minigel format. Minigels provide rapid separation but give lower resolution. Several alternate protocols are provided for specific applications, including electrophoresis of peptides and small proteins, continuous SDS-PAGE, ultrathin gels, multiple single-concentration gels, gradient gels, multiple gradient gels, and multiple gradient minigels. PMID- 18429092 TI - Immunoblot detection. AB - Immunoblotting (often referred to as western blotting) is used to identify specific antigens recognized by polyclonal or monoclonal antibodies. Following electrophoretic separation of proteins and transfer from the gel to an appropriate membrane, the immobilized proteins are probed with specific antibodies to identify and quantitate any antigens present. After being probed with primary antibody, the membrane is washed and the antibody-antigen complexes are identified using horseradish peroxidase (HRPO) or alkaline phosphatase (AP) enzymes coupled to the secondary anti-immunoglobulin-G (anti-IgG) antibody (e.g., goat anti-rabbit IgG). As described in this unit, the detection enzymes are attached directly or via an avidin-biotin bridge to the secondary antibody. Chromogenic or luminescent substrates are also described for visualizing the activity. PMID- 18429093 TI - Autoradiography. AB - This overview provides a thorough discussion of the theory and application of autoradiography for determining the spatial distribution of radioisotope-labeled substances within a membrane or specimen material. The various types of film are discussed along with instructions on their correct handling. Quantitative autoradiography is described as a means of calibrating experimental results. Importantly, specific guidelines are given for pre-flashing film, along with exposure and processing procedures. PMID- 18429094 TI - Capillary electrophoresis of peptides and proteins using isoelectric buffers. AB - Capillary electrophoresis in acidic isoelectric buffers is a novel methodology allowing fast protein and peptide analysis in uncoated capillaries. Due to the low pH adopted and to the use of dynamic coating with cellulose derivatives, silanol ionization is essentially suppressed and no interaction of macromolecules with the untreated wall ensues. In addition, because of the low conductivity of quasi-stationary isoelectric buffers, high voltage gradients can be applied (up to 800 V/cm), permitting fast peptide analysis with a high resolving power and minimal diffusional peak spreading. PMID- 18429095 TI - One-dimensional isoelectric focusing of proteins in slab gels. AB - This unit describes isoelectric focusing (IEF) of polypeptides in urea-containing polyacrylamide slab gels as a result of a pH gradient created by ampholytes (small charged organic molecules) in the gel in response to an electric field. Separation on the basis of isoelectric point produces bands of polypeptides that can be electrotransferred to suitable membranes for further analysis, for example, by staining or immunodetection. If samples have been radiolabeled, they can be visualized by fluorography. This unit contains support protocols detailing special conditions for application of denaturing IEF slab gels for electrotransfer and fluorography in which a scintillator is incorporated into the gel after completion of the run to facilitate detection of radioactively labeled polypeptides. PMID- 18429096 TI - One-dimensional electrophoresis using nondenaturing conditions. AB - Nondenaturing or "native" electrophoresis--i.e., electrophoresis in the absence of denaturants such as detergents and urea--is an often-overlooked technique for determining the native size, subunit structure, and optimal separation of a protein. Two protocols are presented in this unit: continuous PAGE, which is highly flexible, permitting cationic and anionic electrophoresis over a full range of pH, and discontinuous PAGE, which is limited to proteins negatively charged at neutral pH but provides high resolution for accurate size calibration. PMID- 18429097 TI - Two-dimensional gel electrophoresis. AB - Two-dimensional gel electrophoresis combines two different electrophoretic separating techniques in perpendicular directions to provide a much greater separation of complex protein mixtures than either of the individual procedures. Variations of the most common two-dimensional technique are described in this unit, namely isoelectrofocusing (IEF) and SDS-PAGE. This unit also includes support protocols describing pI standards and pH profile measurements, casting Immobiline gels, preparation of tissue culture cells and solid tissues for isoelectricfocusing, preparation of molecular weight standards for two dimensional gels, and two-dimensional protein databases. PMID- 18429098 TI - Electroblotting from polyacrylamide gels. AB - This unit contains procedures for electrophoretically transferring proteins onto a variety of membranes including polyvinylidene difluoride (PVDF) and nitrocellulose, and derivatized membranes. The choice of membrane type for electrotransfer is dependent on the ultimate application for the blot membrane. An alternate protocol is provided for electroblotting in semidry systems. This unit also describes procedures for eluting proteins from membranes using detergents or acidic extraction with organic solvents. PMID- 18429099 TI - Detection of proteins on blot membranes. AB - Staining of blot transfer membranes permits visualization of proteins and allows the extent of transfer to be monitored. In the protocols described in this unit, proteins are stained after electroblotting from one-dimensional or two dimensional polyacrylamide gels to blot membranes such as polyvinylidene difluoride (PVDF), nitrocellulose, or nylon membranes. Protocols are provided for the use of six general protein stains: amido black, Coomassie blue, Ponceau S, colloidal gold, colloidal silver, and India ink. In addition, the fluorescent stains fluorescamine and IAEDANS, which covalently react with bound proteins, are described. Approximate detection limits for each nonfluorescent stain are indicated along with membrane compatibilities. PMID- 18429100 TI - Capillary electrophoresis of proteins and peptides. AB - Capillary electrophoresis (CE) is a high-resolution technique for the separation of a wide variety of molecules of biological interest such as metabolites, drugs, amino acids, nucleic acids, and carbohydrates. This unit focuses on the use of CE to separate proteins and peptides based on charge-to-mass ratios. Separation of proteins based on their isoelectric points is described along with a protocol for optimizing the separation conditions for a given protein. CE is also used for separations of peptides on an analytical scale and as a micropreparative technique--with either multiple separations that are pooled or a single, larger scale separation--for the isolation of peptides from a protease digestion. In most of these examples the same capillary column can be used for all the separations. Only changes in buffer composition, ionic strength, and the presence or absence of additives are required for each specific application. PMID- 18429101 TI - Enzymatic digestion of proteins in solution. AB - Analysis of protein covalent structure is less complex and more accurate when performed on peptides derived from the larger protein. In contrast to acid promoted total hydrolysis, peptides are typically generated by selective proteolysis, i.e., by specifically cleaving peptide bonds with endoproteases that have varying degrees of specificity. This unit presents a protocol that can be used to generate peptide fragments from intact, undenatured proteins. Fragments can be analyzed directly by mass spectrometry (MS) or, more often, are first separated by reversed-phase HPLC (RP-HPLC) and then analyzed by MS or automated sequencing. Most proteins are resistant to enzymatic proteolysis under nondenaturing conditions or are not soluble in aqueous solution. Digestion procedures performed in the presence of chaotropic agents and SDS are described, and support protocols provide instructions for preparing enzyme stocks and reducing and alkylating peptides prior to sequencing or HPLC analysis. PMID- 18429102 TI - N-terminal sequence analysis of proteins and peptides. AB - Amino-terminal (N-terminal) sequence analysis is used to identify the order of amino acids of proteins or peptides, starting at their N-terminal end. This unit describes the sequence analysis of protein or peptide samples in solution or bound to PVDF membranes using a Perkin-Elmer Procise Sequencer. Sequence analysis of protein or peptide samples in solution or bound to PVDF membranes using a Hewlett-Packard Model G1005A sequencer is also described. Methods are provided for optimizing separation of PTH amino acid derivatives on Perkin-Elmer instruments and for increasing the proportion of sample injected onto the PTH analyzer on older Perkin-Elmer instruments by installing a modified sample loop. The amount of data obtained from a single sequencer run is substantial, and careful interpretation of this data by an experienced scientist familiar with the current operation performance of the instrument used for this analysis is critically important. A discussion of data interpretation is therefore provided. Finally, discussion of optimization of sequencer performance as well as possible solutions to frequently encountered problems is included. PMID- 18429103 TI - Enzymatic digestion of proteins on PVDF membranes. AB - Enzymatic digestion of membrane-bound proteins is one of the most widely used procedures for determining the internal amino acid sequence of proteins that either have a blocked amino terminus or require two or more stretches of sequence data for DNA cloning or confirmation of protein identification. Because the final step of protein purification is usually SDS-PAGE, electroblotting to either polyvinylidene difluoride (PVDF) or nitrocellulose is the simplest and most common procedure for recovering protein free of contaminants (e.g., SDS or acrylamide) with a high yield. As described in this unit, PVDF is preferred over nitrocellulose because it can be used for a variety of other structural analysis procedures, such as amino-terminal sequence analysis and amino acid analysis. In addition, peptide recovery from PVDF membranes is higher than from nitrocellulose, particularly from higher-retention PVDF (e.g., ProBlott, Transblot, Westran, or Immobilon P(sp)). Finally, PVDF-bound protein can be stored dry, as opposed to nitrocellulose-bound protein, which must remain wet during handling and storage to prevent loss of peptides during digestion. PMID- 18429104 TI - Chemical cleavage of proteins on membranes. AB - Described in this unit are five basic protocols that are widely used for specific and efficient chemical cleavage of proteins bound to membranes. Cyanogen bromide (CNBr) cleaves at methionine (Met) residues; BNPS-skatole cleaves at tryptophan (Trp) residues; formic acid cleaves at aspartic acid-proline (Asp-Pro) peptide bonds; hydroxylamine cleaves at asparagine-glycine (Asn-Gly) peptide bonds, and 2 nitro-5-thiocyanobenzoic acid (NTCB) cleaves at cysteine (Cys) residues. Because the above loci are at relatively low abundance in most proteins, digestion with these agents will yield relatively long peptides. In addition, Alternate Protocol an describes CNBr cleavage of PVDF-bound protein previously analyzed by Edman degradation. Finally, a Support Protocol discusses preferred methods of separating and analyzing peptide fragments generated by the chemical cleavage reactions described in the basic protocols. PMID- 18429105 TI - Reversed-phase isolation of peptides. AB - Reversed-phase high-performance liquid chromatography (HPLC) is a fundamental tool for the isolation and analysis of peptides. Peptides are separated on a hydrophobic stationary phase and eluted with a gradient of increasing organic solvent concentration. This unit presents protocol for separation of 5- to 500 pmol of peptides on a narrow-bore (2-mm-i.d.) or microbore (1-mm-i.d.) column. Smaller quantities can be separated capillary HPLC columns, as described. Capillary HPLC columns, however, require a gradient flow rate of 3 to 5 ml/min, which most current HPLC pumps cannot attain without modifications. A procedure is therefore provided for constructing a capillary HPLC system using readily available components. HPLC peaks that appear to be symmetrical may actually contain coeluting peptides. Matrix-assisted laser desorption/ionization (MALDI) mass spectrometry and capillary electrophoresis are described for analysis of a small portion of an HPLC fraction to determine the number of components present in a small sample. These methods can be utilized to screen fractions prior to automated sequencing. PMID- 18429106 TI - Removal of N-terminal blocking groups from proteins. AB - Two enzymatic methods commonly used in N-terminal sequence analysis of blocked proteins are presented in this unit; one uses pyroglutamate aminopeptidase for N(alpha)-pyrrolidone carboxyl-proteins in solution or blotted onto a membrane, and the other uses acylaminoacyl-peptide hydrolase for N(alpha)-acyl-proteins blocked with other acyl groups. A Support Protocol describes a colorimetric assay for pyroglutamate aminopeptidase activity. Sequencing with acylaminoacyl-peptide hydrolase must include fragmentation of the protein before unblocking can be carried out, so procedures are provided for chemically blocking newly generated peptides with either succinic anhydride or phenylisothiocyanate/performic acid. The hydrolase is then applied to the total mixture of peptides, only one of which, the acylated N-terminal peptide, should be a substrate for hydrolase. After incubation, the mixture of peptides is subjected to sequence analysis. Protocols are also provided for unblocking N-terminally blocked proteins using acid-catalyzed hydrolysis or methanolysis, hydrazinolysis, and beta-elimination after acid-catalyzed N-O shift. Alternate protocols describe chemical removal of acetyl and longer-chain alkanoyl groups, as well as formyl groups to open the cyclic imide of pyrrolidone carboxylate. PMID- 18429107 TI - Amino acid analysis. AB - Amino acid analysis (AAA) is one of the best methods to quantify peptides and proteins. Two general approaches to quantitative AAA exist, namely, classical postcolumn derivatization following ion-exchange chromatography and precolumn derivatization followed by reversed-phase HPLC (RP-HPLC). Excellent instrumentation and several specific methodologies are available for both approaches, and both have advantages and disadvantages. This unit focuses on picomole-level AAA of peptides and proteins using the most popular precolumn derivatization method, namely, phenylthiocarbamyl amino acid analysis (PTC-AAA). It is directed primarily toward those interested in establishing the technology with a modest budget. PTC derivatization and analysis conditions are described, and support and alternate protocols describe additional techniques necessary or useful for most any AAA method--e.g., sample preparation, hydrolysis, instrument calibration, data interpretation, and analysis of difficult or unusual residues such as cysteine, tryptophan, phosphoamino acids, and hydroxyproline. PMID- 18429108 TI - Overview of glycoconjugate analysis. AB - Whereas DNA, RNA, and proteins are linear polymers that can usually be directly sequenced, oligosaccharides show substantially more complexity,having branching and anomeric configurations (alpha and beta linkages). The biosynthesis of oligosaccharides, termed glycosylation, is extremely complex, is not template driven, varies among different cell types, and cannot be easily predicted from simple rules. This overview discusses the stereochemistry of mono- and oligosaccharides and provides diagrammatic representations of monosaccharides (Fisher projections and Haworth representations) and formulas for representation of oligosaccharide chains. A glossary of terms used in glycobiology is also provided. PMID- 18429109 TI - Metabolic radiolabeling of animal cell glycoconjugates. AB - Useful information about glycoconjugates can be obtained by labeling their aglycone (noncarbohydrate) portions--e.g., labeling proteins with radioactive amino acids-and then using techniques described elsewhere in this chapter to infer the presence, type, and nature of oligosaccharide chains. This unit describes metabolic labeling techniques that provide more specific information about the structure, sequence, and distribution of the sugar chains of glycoconjugates. Following metabolic labeling, the radioactive glycoconjugate of interest is isolated, individual glycosylation sites are identified and separated if necessary, and the labeled oligosaccharides are subjected to structural analysis. PMID- 18429110 TI - Inhibition of N-linked glycosylation. AB - This unit describes the use of inhibitors in cultured cells to prevent N-linked glycosylation of proteins to yield glycoproteins containing missing or altered chains. This approach is useful for examining potential functional role(s) of oligosaccharides on specific proteins or intact cells. First, the optimal concentration of inhibitor for the experiment (i.e., highest nontoxic concentration) is determined by monitoring [(35)S]methionine incorporation as a measure of protein biosynthesis. The inhibitor's ability to inhibit oligosaccharide processing is then determined by analyzing cells labeled with [(3)H]mannose using TCA precipitation or endo H digestion. A support protocol details a method for concentrating proteins by acetone precipitation. PMID- 18429111 TI - Detection of glycophospholipid anchors on proteins. AB - Many eukaryotic proteins are tethered to the plasma membrane by glycosyl phosphatidylinositol (GPI) membrane anchors. This unit provides a general approach for detecting GPI-anchored proteins. First, the detergent-partitioning behavior of a protein of interest is examined for characteristics of GPI-linked species. The partitioning of total cellular and isolated proteins with Triton X 114 is described in this unit, and precondensation of Triton X-114, which is necessary to remove hydrophilic contaminants before partitioning, is outlined in a Support Protocol 1. The protein may also be subjected to specific enzymatic or chemical cleavages to release it from its GPI anchor. Phospholipase cleavage (starting with intact cells or membranes, or with isolated protein) is detailed, and chemical cleavage with nitrous acid is also described. If GPI-anchored proteins are radiolabeled with fatty acids, it facilitates the detection of the GPI protein products following the cleavage reactions. A protocol for separation of lipid moieties released from proteins is provided and base hydrolysis of proteins is also presented. PMID- 18429112 TI - Detection and analysis of proteins modified by O-linked N-acetylglucosamine. AB - First, a protocol for increasing the stoichiometry of O-GlcNAc on proteins is given. This is followed by simple techniques for the detection/screening of O GlcNAc-modified proteins either by immunoblotting or lectin affinity chromatography. Separate protocols verify that the glycan is O-linked GlcNAc. These methods are followed by protocols for more comprehensive analysis of O GlcNAc modified proteins, including labeling of O-GlcNAc residues with [3H]Gal, and subsequent product analysis. The final two protocols assay for O-GlcNAc transferase and O-GlcNAcase activity, respectively. PMID- 18429113 TI - Overview of protein phosphorylation. AB - This overview provides a history of protein phosphorylation research and provides the reader with an understanding of how and why labeling studies are performed. The various sites of protein phosphorylation are described along with the roles of the many kinases and phosphatases that regulate phosphorylation. Methods for detecting unlabeled phosphoamino acids, including high-voltage electrophoresis on thin-layer cellulose acetate plates, gel-shift assays, and the use of anti phosphopeptide antibodies. PMID- 18429114 TI - Labeling cultured cells with 32P(i) and preparing cell lysates for immunoprecipitation. AB - This unit describes (32)P(I) labeling and lysis of cultured cells to be used for subsequent immunoprecipitation of proteins. The procedure is suitable for insect, avian, and mammalian cells and can be used with both adherent and nonadherent cultures. The protocol described is (32)P(I) labeling of adherent or nonadherent (e.g., hematopoietic) cells with subsequent lysis in a detergent buffer. More rigorous lysis conditions to be used for working with proteins that are difficult to solubilize are also described. PMID- 18429115 TI - Phosphoamino acid analysis. AB - It is often valuable to identify the phosphorylated residue in a protein. This unit presents a protocol for partial acid hydrolysis of proteins phosphorylated at serine, threonine, or tyrosine, followed by two-dimensional thin-layer electrophoresis of the labeled phosphoamino acid. Phosphothreonine and phosphotyrosine are more stable to hydrolysis in alkali than are RNA and phosphoserine. Therefore, an alternate procedure using mild alkaline hydrolysis of protein samples to enhance the detection of phosphothreonine and phosphotyrosine is also provided. PMID- 18429116 TI - Detection of phosphorylation by immunological techniques. AB - Phosphorylation of unlabeled proteins can be detected using immunological or enzymatic techniques. Anti-phosphotyrosine antibodies are used with immunoblots to detect tyrosine phosphorylation. This unit presents a protocol employing anti phosphotyrosine antibodies with detection by either (125)I-labeled protein A or enhanced chemiluminescence (ECL). PMID- 18429117 TI - Detection of phosphorylation by enzymatic techniques. AB - This unit presents protocols for examining the functional effects elicited by nonspecific acid or alkaline phosphatases that dephosphorylate many phosphoproteins in vitro. Additional protocols describe digestion of phosphoproteins with a protein serine/threonine phosphatase and protein tyrosine phosphatase. A support protocol has been included to identify the radiolabel as (32)P(I) based on its ability to form a complex with ammonium molybdate. PMID- 18429118 TI - Assays of protein kinases using exogenous substrates. AB - In studies of the regulation of specific biochemical events by reversible phosphorylation, assaying the protein kinases themselves can often lead to significant progress in understanding the mechanistic details of a system under study. This unit describes assays for a variety of protein kinases that require different conditions to detect and measure their activities--cyclic nucleotide dependent kinases, protein kinase C and isoforms, casein kinases, Ca(2+)/calmodulin-dependent kinases, and tyrosine kinase. The unit is not meant to be a catalog of individual protein kinase assays; however, the general principles of these assays should apply to most if not all known protein kinases. In-gel assays for specific kinase activity are also described. Support protocols describe production of crude cell lysates as a source of enzyme activity, and methods of evaluating assays, including trichloroacetic acid (TCA) precipitation to measure incorporation of radioactivity, adsorption of the labeled material onto P81 phosphocellulose paper, or electrophoretic separation and autoradiography. PMID- 18429119 TI - Permeabilization strategies to study protein phosphorylation. AB - This unit deals with the use of nucleotide triphosphates to label proteins in vitro in permeabilized cells and isolated cellular fractions. Both of these assay formats result in lysates from which the protein of interest may be easily immunoprecipitated; however alternative techniques are described for preparing the final lysate for electrophoretic analysis. A related procedure that does not involve permeabilization is outlined for direct analysis of cytosolic or membrane bound kinases. Two different methods for determining the specific radioactivity of (32)P-containing compounds are also included. These experiments generally utilize [gamma-(32)P]ATP as an exogenously added phosphate donor, although [gamma (32)P]GTP can be used in specific cases. The method is very straightforward, although numerous considerations must be made before applying it to each new system. PMID- 18429120 TI - Phosphopeptide mapping and identification of phosphorylation sites. AB - Protein phosphorylation is a common modification for many proteins in the cell. Phosphorylation can affect localization of a protein, its stability, and its ability to dimerize or form stable complexes with other molecules. To understand the underlying mechanisms behind the phosphorylation of a given protein, it is often necessary to precisely identify which amino acid residues are phosphorylated. This unit describes the technique of phosphopeptide mapping. In this procedure, a radiolabeled protein is proteolytically digested, and the resulting phosphopeptides are separated in two dimensions on a TLC plate. The phosphopeptides are also analyzed by HPLC and mass spectrometry or peptide microsequencing. Such mapping gives information about the number of phosphorylation sites present in the protein, and can also be used to find out if sites of phosphorylation on a protein change upon treatment of cells with specific agents. PMID- 18429121 TI - Analysis of disulfide bond formation. AB - In this unit, protocols are provided for detection of disulfide bond formation in cultures of intact cells and in an in vitro translation system containing isolated microsomes. First, the newly synthesized protein of interest is biosynthetically labeled with radioactive amino acids in a short pulse. The labeled protein is then chased with unlabeled amino acids. At different times during the chase, a sample is collected, membranes are lysed with detergent, and the protein is isolated by immunoprecipitation, as described. A support protocol is provided for analysis of disulfide bonds in the immunoprecipitates by SDS-PAGE with and without prior reduction. The difference in mobility observed between the gels with unreduced and reduced samples is due to disulfide bonds in the unreduced protein. PMID- 18429122 TI - Analysis of protein acylation. AB - Protein acylation is the covalent attachment of fatty acids to a protein; the most commonly added fatty acids are myristate (14:0) and palmitate (16:0). In this unit, protocols describe the use of radiolabeled fatty acids to label eukaryotic cells in vitro. The radiolabeled material produced can then be analyzed by the various methods described here: determination of the type of fatty acid linkage, checking for interconversion by determining the nature of the protein-bound label, and identification of the protein-bound fatty acid. PMID- 18429123 TI - Analysis of protein prenylation and carboxyl-methylation. AB - This unit describes methods for analysis of prenylation and the carboxyl methylation that often accompanies it. The two prenoid groups that have been found attached to proteins--farnesyl (C15) and geranylgeranyl (C20)--are both derived from intermediates in the isoprenoid biosynthetic pathway that utilizes mevalonic acid. In the protocols described here, radiolabeled mevalonate is used to label these intermediates in either intact cells or in vitro; the labeled intermediates then become incorporated into proteins. Alternatively, the preformed radioactive prenyl diphosphates can be used for in vitro translations, as described here. Carboxyl-methylation of C-terminal prenylated cysteine residues often occurs subsequent to prenylation. Methods are given for radiolabeling of the methyl group with [(3)H-methyl]methionine, that is converted intracellularly into S-adenosylmethionine, and for radiolabeling with preformed S adenosyl[(3)H]methionine. PMID- 18429124 TI - Analysis of oxidative modification of proteins. AB - Protein oxidation has been implicated in a variety of degenerative diseases as well as in the aging process. This unit describes techniques for the quantification of various protein oxidation products, including protein carbonyls, loss of protein thiol groups, dityrosine and nitrotyrosine, and isoaspartate formation. Such oxidatively modified products may also be used as biomarkers for the assessment of oxidative stress during aging and/or disease. PMID- 18429125 TI - Modification of cysteine. AB - This unit describes a number of methods for modifying cysteine residues of proteins and peptides by reduction and alkylation procedures. A general procedure for alkylation of cysteine residues in a protein of known size and composition with haloacyl reagents or N-ethylmaleimide (NEM) is presented, and alternate protocols describe similar procedures for use when the size and composition are not known and when only very small amounts of protein are available. Alkylations that introduce amino groups using bromopropylamine and N-(iodoethyl) trifluoroacetamide are also presented. Two procedures that are often used for subsequent sequence analysis of the protein, alkylation with 4-vinylpyridine and acrylamide, are described, and a specialized procedure for 4-vinylpyridine alkylation of protein that has been adsorbed onto a sequencing membrane is also presented. Reversible modification of cysteine residues by way of sulfitolysis is described, and a protocol for oxidation with performic acid for amino acid compositional analysis is also provided. Gentle oxidation of cysteine residues to disulfides by exposure to air is detailed. Support protocols are included for recrystallization of iodoacetic acid, colorimetric detection of free sulfhydryls, and desalting of modified samples. PMID- 18429126 TI - Modification of amino groups. AB - This unit describes group-specific modifications of amino groups. These reactions remain valid tools for early-stage evaluation of structure-function relationships, but are now valued even more for their applications in the preparation of bioconjugates, affinity columns, biosensors, and tagged macromolecules. Protocols are provided here for reaction of amino groups with succinimidyl esters and isothiocyanates. These methods are broadly useful for the stable coupling to proteins of groups with useful, non-native functional properties. These include biotin for detection or recovery, fluorescent groups for biophysics or cytochemistry, cross-linking reagents for making bioconjugates, or metal-chelators that permit proteins to be loaded with radioisotopes for medical imaging or antitumor therapy. These applications require accurate product characterization, which preferably is performed by mass spectrometry, as described in this unit as a support procedure. A protocol employing succinic or acetic anhydrides to change the charge state of protein amino groups is provided here, as is a procedure for reductive alkylation that leaves their charge unaltered but converts primary amines to secondary or tertiary amines. PMID- 18429127 TI - Overview of peptide and protein analysis by mass spectrometry. AB - One goal of this introductory overview is to try and dispel the still widely held, but now mistaken, belief that mass spectrometry is beyond the technical resources of the typical laboratory or facility engaged in the structural characterization or synthesis of peptides and proteins. Over the last five years, mass spectrometers and associated data systems have become available that can be operated by anyone capable of running a modern amino acid analyzer or Edman sequencer, yet are powerful enough to handle the most demanding analyses that the peptide or protein investigator might require. The remaining goals of this article are to familiarize peptide and protein chemists with the types of mass spectrometers that are appropriate for the majority of their analytical needs, to describe the kinds of experiments that can be performed with these instruments on a routine basis, and to discuss the kinds of information that these experiments provide. The emphasis here is on established tools and techniques that can realistically be used for problem solving. As a result, many useful instruments and techniques employed by the trained mass spectroscopist are not discussed here, because they do not satisfy these criteria or are not yet widely available; however, a few of these are briefly mentioned at the end of the unit. The overview concludes with a tutorial discussion on the meaning and practical importance of a number of fundamental experimental and performance-related issues that have important implications for interpretation and use of mass spectral data, such as mass accuracy and resolution. PMID- 18429128 TI - Protein identification using a quadrupole ion trap mass spectrometer and SEQUEST database matching. AB - A procedure is described for rapid, sensitive protein identification utilizing liquid chromatography--tandem mass spectrometry. The analysis is performed on mixtures of peptides obtained by enzyme digestion. The SEQUEST computer program is used to match the sequence information in the spectra to a database of known protein sequences. PMID- 18429129 TI - Matrix-assisted laser desorption/ionization time-of-flight mass analysis of peptides. AB - Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) is one of the most useful techniques for determining the mass of biomolecules, with exceptional capabilities for mass analysis of peptides. Relative to other ionization techniques, it provides high sensitivity and excellent tolerance of salt and other common buffer components. Routine detection limits for peptides are in the subpicomole range. The ions commonly observed are the protonated molecules (M+H(+)), which makes data analysis relatively easy. This overview discusses instrument configuration and calibration, sample preparation, along with specific approaches for analyzing peptide mixtures, synthetic peptides, and chemical modifications of peptides. PMID- 18429131 TI - In-gel digestion of proteins for MALDI-MS fingerprint mapping. AB - Mass spectrometry (MS) has emerged as a sensitive, versatile, and rapid method for protein identification, following the advent of electrospray ionization mass spectrometry (ESI-MS) and matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). The advantages of MALDI-MS over ESI-MS include its relatively high tolerance to contamination from biological matrices, its high sensitivity, the relative ease of interpreting spectra from mixtures, and the formation of singly protonated molecular ions for tandem analysis. Peptide fingerprint mass mapping and partial peptide sequencing using post-source decay and ladder sequencing by MALDI-MS in combination with algorithms for sequence database interrogation have the potential for identification and structural investigation of proteins. This unit describes in-gel digestion for peptide mass mapping of picomole to subpicomole quantities of protein derived from Coomassie- or silver-stained polyacrylamide gels. After digestion, the peptides are extracted from the gel and mass analyzed. PMID- 18429130 TI - Sample preparation for MALDI mass analysis of peptides and proteins. AB - Sample preparation may be the most crucial step in mass spectrometric analysis of peptides and proteins by MALDI. The free dipolar ionic peptides or proteins must be incorporated into a crystalline solid solution that is comprised of excess solvent (the matrix), which modulates desorption/ionization by preferentially absorbing the laser energy at the irradiation wavelength and laser pulse intensity being employed. Usually the cocrystalline sample and matrix are prepared by mixing solutions of each component and permitting the mixture to crystallize through evaporation of the solvents. Two methods employing this approach are presented in this unit: the dried drop method and the rapid evaporation method. PMID- 18429132 TI - Searching sequence databases over the internet: protein identification using MS Fit. AB - Peptide fingerprint mass mapping is widely used for initial identification of proteins separated by gel electrophoresis. This method involves the enzymatic in gel digestion of proteins to generate peptides, followed by mass measurement of the cleaved peptides. The experimental mass values are then compared with theoretical values from protein databases, calculated using the cleavage specificity of the enzyme. This unit describes the use of the search program MS Fit to identify the closest match or matches. PMID- 18429133 TI - Searching sequence databases over the internet: protein identification using MS tag. AB - Although peptide mass mapping is fast and simple, its success can be compromised by the purity of the protein in one gel spot, errors in the sequence database, and the detection of too few peptides in the MS map of a given protein to permit reliable database matching. In those cases, partial amino acid sequence that is determined using post-source decay (PSD) analysis or tandem mass spectrometry is required to establish the correct protein identification among possibilities suggested by MS-Fit. Post-source decay (PSD) involves the detection of the fragmentation product ions (metastable ions) of a selected mass value window containing the precursor ion chosen, that occur in the field-free region between the ion source and the reflectron. This unit covers some of the practical issues in obtaining and analyzing a typical PSD spectrum using MS-Tag, a database interrogation program that enables the identification of proteins with a limited number of peptide fragment ions by matching existing database sequences. It is a user-friendly program that simultaneously considers a variety of ion types, but still allows single-mismatch-tolerant searching. Although MALDI-PSD database searching is a widely used tool, an alternative method also described in this unit is analysis by MALDI collision-induced dissociation (MALDI-CID). PMID- 18429134 TI - Enzymatic approaches for obtaining amino acid sequence: on-target ladder sequencing. AB - Peptide sequencing by mass spectrometry (MS) is usually based on detecting mass differences associated with various amino acids in the polymer chain. Post-source decay (PSD) and MS/MS spectra may yield internal peptide sequences. However, determination of the order of the first two, and sometimes the last few, amino acids in the peptide is often problematic without additional experiments. Several enzymatic approaches have proven useful for identifying the N- and C-terminal residues. They involve the use of carboxypeptidases and aminopeptidases to produce peptide ladders for rapid analysis by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). This unit describes the ladder sequence method to generate amino acid sequence information from low- to subpicomole quantities of peptides. It can be performed directly on the sample stage, thus minimizing potential sample losses to vials and through sample transfer. PMID- 18429135 TI - Introducing samples directly into electrospray ionization mass spectrometers using a nanospray interface. AB - Electrospray (ES) is a concentration-dependent phenomenon, and operation at high flow rates results in a corresponding high rate of sample consumption. Sensitivity increases are most readily achieved by decreasing the flow rate of sample solution into the electrospray source. In recent years, a number of methodologies have been developed to achieve ES analyses at sub-ml/min flow rates. There are a large number of microscale ES devices currently in use, but this unit describes in detail a micro-scale interface for direct liquid introduction without separation. PMID- 18429136 TI - Introducing samples directly into electrospray ionization mass spectrometers using microscale capillary liquid chromatography. AB - This unit describes the design and operation of a microscale electrospray (ES) interface suitable for the on-line liquid chromatography (LC) separation and mass spectrometry (MS) analysis of mixtures of peptides and proteins. The interface utilizes an ES needle packed with reversed-phase support. Such a design has the advantage of minimizing any void volume between the end of the column and point of electrospray ionization, thus maintaining the integrity of the LC separation and maximizing sensitivity. Here, protocols are presented for construction of an integrated LC column ES needle in-house, packing the ES needle, and mounting and using the microscale ES LC/MS interface assembly. Various options for low-flow solvent delivery systems are also discussed. PMID- 18429137 TI - Principles of macromolecular X-ray crystallography. AB - X-ray crystallography is one of the major tools available for protein structural analysis. This unit provides an introductory review of the principles of X-ray crystallography that covers how the image is generated and analyzed. The second half of the unit describes strategies for producing crystals of protein, including methods for dealing with proteins that do not easily form crystals. PMID- 18429138 TI - Introduction to NMR of proteins. AB - This unit aims to overview the application of NMR spectroscopy to proteins. It is not intended to provide an exhaustive "how to" guide, but rather to give a flavor of both well-established and emerging NMR techniques used in the elucidation of protein structure. It starts with a brief introduction to the basic principles of NMR and the information provided by this technique, and goes on to discuss the instrumentation involved, spectral assignment methods for small and large proteins, and the utility of other spin active nuclei (e.g., (13)C and (15)N) to aid assignment of the latter. PMID- 18429139 TI - Synthesis of multiple peptides on plastic pins. AB - This unit covers the strategy of the multiple peptide approach to biological scanning, the synthetic protocols, and the handling of peptides after synthesis: cleavage, preliminary purification, storage, and analysis. It is specific for the multipin technique using equipment obtained from Chiron Technologies, although some of the approaches are applicable to other multiple synthesis techniques. This unit also includes protocols for preparing Fmoc-amino acid solutions and for acetylating or biotinylating synthesized peptides. PMID- 18429140 TI - Native chemical ligation of polypeptides. AB - The total synthesis and semisynthesis of proteins allows the site-specific incorporation of unnatural amino acids, post-translational modifications, and biophysical/biochemical probes into the target molecule. Among the various chemical and enzymatic approaches available for the synthesis/semisynthesis of proteins, the native chemical ligation technique has proven especially useful and is the exclusive focus of this unit. This unit first discusses how to choose the ligation site(s) in the target protein and then outlines how to obtain the necessary polypeptide building blocks using either chemical synthesis or recombinant DNA expression. Next, the synthesis of a protein by native chemical ligation of two polypeptide fragments is described. The synthesis of a protein from three polypeptide fragments using a sequential native chemical ligation strategy is also described. Support protocols describe how to obtain the necessary polypeptide fragments using either chemical synthesis or recombinant DNA expression. PMID- 18429141 TI - Synthesis and application of peptide dendrimers as protein mimetics. AB - The use of peptides to mimic a portion of a protein structure is a challenging and powerful tool in the discovery of new drugs. In native proteins, discontinuous bioactive peptide surfaces are held together in a particular conformation by the structural rigidity of the protein. Approaches to mimicking a structural surface center on bringing the potential peptide sequences together by assembling the peptide chains on a template. These templates can be flexible dendrimeric or cyclic peptides as well as more rigid organic molecules. The Multiple Antigen Peptide (MAP) system represents a novel approach to preparing peptide immunogens. The MAP consists of an inner core matrix built up of a large layer of Lys residues and a surface of peptide chains attached to the core matrix. Because of its dendrimeric structure, MAP can be very useful as a template for assembling potential peptide surfaces. A variation of this procedure, the cyclic Multiple Antigen Peptide (cMAP) approach, is also presented here. Having branched multiple closed-chain architectures, the cMAP system is often a superior approach for protein mimetics because the multiple constrained peptides can mimic bioactive conformations. Whether to select this approach over MAP depends on the properties of the peptides, but usually if the peptides are too small to adopt a stable conformation on their own, incorporation of a cyclic structure may be necessary. MAPs have been applied to areas of study such as inhibitors, artificial proteins, affinity purifications, and intracellular transport. PMID- 18429142 TI - Disulfide bond formation in peptides. AB - The formation of disulfide bridges is often a crucial final stage in peptide synthesis. There is compelling evidence that the disulfide pattern can be critical in the folding and structural stabilization of many natural peptide and protein sequences, while the artificial introduction of disulfide bridges into natural or designed peptides may often improve biological activities/specificities and stabilities. This unit provides a highly selective, albeit state-of-the-art, menu of procedures that can be performed to establish intramolecular or intermolecular disulfide bridges in targets of varying complexities. PMID- 18429143 TI - Analysis of protein-protein interactions. AB - This overview defines some of the terminology used in the study of protein protein interactions and also describes the basis of equilibrium kinetic assays (i.e., defining necessary assumptions, terms and equations). PMID- 18429144 TI - Interaction trap/two-hybrid system to identify interacting proteins. AB - The parameters and experimental conditions for this important system are constantly undergoing improvement. This newest version includes expanded tables describing interaction trap components and additional libraries compatible with the interaction trap system. It also features a new protocol on performing a hunt by interaction mating. Some of the commercial vendors selling yeast two-hybrid reagents recommend using interaction mating to perform a hunt, so this procedure should be of great interest. PMID- 18429145 TI - Phage-based expression cloning to identify interacting proteins. AB - Interaction cloning (also known as expression cloning) is a technique to identify and clone genes which encode proteins that interact with a protein of interest, or "bait" protein. The procedure presented in this unit involves a fusion protein consisting of bait protein and glutathione-S-transferase (GST) with a protein kinase A site at the junction between them (the protocol can, however, be adapted to use other PKA-containing recombinant proteins). The labeled protein is subsequently used as a probe to screen a l bacteriophage-derived cDNA expression library, which expresses b -galactosidase fusion proteins that contain in-frame gene fusions. The phages lyse cells, form plaques, and release fusion proteins that are adsorbed onto nitrocellulose membrane filters. The filters are blocked with excess nonspecific protein to eliminate nonspecific binding and probed with the radiolabeled bait protein. This procedure leads directly to the isolation of genes encoding the interacting protein, bypassing the need for purification and microsequencing or for antibody production. PMID- 18429146 TI - Imaging protein-protein interactions by fluorescence resonance energy transfer (FRET) microscopy. AB - Detection of specific protein-protein interactions has long been restricted to bulk biochemical methods such as immunoprecipitation and immunoblotting. Even more sensitive methods using general immunofluorescence are limited, and it is difficult to infer protein-protein interactions from the results of these tests. Fluoresence Resonance Energy Transfer (FRET) is a photophysical process that can be exploited to obtain highly sensitive information about such interactions. It can sense the presence of acceptor fluorophores in the vicinity of a donor fluorophore within a separation distance that is the size of a single protein molecule. This unit details FRET microscopy based on release of quenched donor fluorescence after acceptor photobleaching, microinjection of reagents into the nucleus or cytosol, and labeling of antibodies for these procedures. PMID- 18429147 TI - High-throughput screening for protein-protein interactions using yeast two-hybrid arrays. AB - Arrays are used for parallel assays of large numbers of proteins in a single experiment. They provide an alternative to libraries for protein screening purposes. This unit describes a protein array composed of living yeast (Saccharomyces cerevisiae) cells that can be used in a functional screen for protein interactions, the two-hybrid assay. The protocols described here use the yeast proteome as an example, but they can be applied to any other genome or subset thereof. PMID- 18429148 TI - Identification of protein interactions by far western analysis. AB - This unit describes far western blotting, a method of identifying protein-protein interactions. In a far western blot, one protein of interest is immobilized on a solid support membrane, then probed with a non-antibody protein. As described, far western blots can be used to identify specific interacting proteins in a complex mixture of proteins. They are particularly useful for examining interactions between proteins that are difficult to analyze by other methods due to solubility problems or because they are difficult to express in cells. This method is performed totally in vitro, and the proteins of interest can be prepared in a variety of ways. A protocol is also provided for determining the effects of specific peptide residues or post-translational modifications on protein-protein interactions. Many different detection techniques, either radioactive or nonradioactive, can be used. For example, the protein probe may be detected indirectly with an antibody, rather than being labeled radioactively. PMID- 18429149 TI - Computational methods for protein sequence analysis. AB - This unit is presented as a guide to addressing the issue of what to do with a protein sequence once it is obtained. A theoretical background for protein sequence analysis is provided first, followed by a discussion of matrix methods for sequence comparison (Matrix Methods for Sequence Comparison: Dot Plots). Sequence similarity searching is then presented, including the BLAST and FASTA databases. Other aspects of protein sequence analysis covered here are alignment methods, scoring matrices, multiple alignments, cluster methods and trees, and identification of functional sites. PMID- 18429150 TI - Hydrophobicity profiles for protein sequence analysis. AB - Hydrophobic interactions are a major force in protein folding and numerous hydropathy scales have been developed to quantify the relative hydrophobicity of the amino acids. Hydropathy profiles can be used to examine the surface features of proteins in order to generate hypotheses that can be confirmed experimentally. This unit describes the application of hydrophobicity plots to typical problems and provides suggested uses for a few selected scales. PMID- 18429151 TI - Protein secondary structure prediction. AB - This unit describes procedures developed for predicting protein structure from the amino acid sequence. The first of the four sections is an overview and brief history of structure prediction schemes. The second section describes four distinct prediction schemes, with emphasis on their differences. In the third part each prediction scheme is used to evaluate three proteins that have different folding patterns. The final section is a comparison of the prediction results and suggestions for secondary structure prediction. PMID- 18429152 TI - Internet basics. AB - With the explosion of sequence and structural information available to researchers, the field of bioinformatics is playing an increasingly large role in the study of fundamental biomedical problems. The challenge facing computational biologists will be to aid in gene discovery and in the design of molecular modeling, site-directed mutagenesis, and experiments of other types that can potentially reveal previously unknown relationships with respect to the structure and function of genes and proteins. This challenge becomes particularly daunting in light of the vast amount of data that has been produced by the Human Genome Project and other systematic sequencing efforts to date. This unit begins with a review of the Internet and its terminology, also discussing major classes of Internet protocols, without becoming overly engaged in the engineering minutiae underlying these protocols. Matters of connectivity, ranging from simple modem connections to digital subscriber lines (DSL) are also discussed. Finally, one of the most common problems that has arisen with the proliferation of Web pages throughout the world is addressed--i.e., finding useful information on the World Wide Web. PMID- 18429153 TI - Sequence similarity searching using the BLAST family of programs. AB - The BLAST (Basic Local Alignment Search Tool) family of sequence similarity search programs allows users to input either a nucleotide or amino acid query sequence, and search a nucleotide or amino acid sequence database. The program returns a list of the sequence "hits", alignments to the query sequence, and statistical values. This unit describes how to choose an appropriate BLAST program and database, perform the search, and interpret the results. PMID- 18429154 TI - Protein tertiary structure prediction. AB - This unit addresses how to predict the tertiary structure of a protein from its amino acid sequence using computational methods. Three types of prediction methods--homology modeling, fold recognition, and ab initio prediction--are introduced. PMID- 18429155 TI - Protein tertiary structure modeling. AB - Insights into the 3D-structure of a protein have proven useful during experiment design. Experimentally elucidated structures are often not available, but comparative protein modeling provides a viable alternative in many cases. This unit presents comparative protein modeling and how to use the highly sophisticated but easy-to-use free software available on the Internet. PMID- 18429156 TI - Overview of the quantitation of protein interactions. AB - The biological function of many proteins involves reversible interactions with other proteins, nucleic acids, or other non-protein ligands. Such interactions play many different roles in a wide range of cellular processes. A few examples are: (1) storing or transporting key metabolites (e.g., O(2) storage by myoglobin); (2) forming and maintaining the quaternary structure of multi-subunit enzymes; (3) specific binding and recognition events (antigen-antibody, hormone receptor, transcription factor-promoter); and (4) self-assembly of large structures (microtubules, chromatin). Thus, the quantitative characterization of such interactions represents an important part of understanding the function of such proteins and their role in these cellular events. This unit sets the tone for the rest of the chapter, and gives important information necessary to understand some of the topics that will be covered in future supplements, such as sedimentation equilibrium (analytical and micro-preparative), surface plasmon resonance (SPR), size-exclusion chromatography (SEC) with on-line light scattering, and chemical cross-linking. PMID- 18429157 TI - Measuring protein interactions by optical biosensors. AB - This unit gives an introduction to the basic techniques of optical biosensing for measuring equilibrium and kinetics of reversible protein interactions. Emphasis is given to the description of simple and robust approaches that will provide reliable results. How to avoid the most commonly encountered problems and artifacts is also discussed. PMID- 18429158 TI - Analytical centrifugation: equilibrium approach. AB - The specific interaction of biological molecules with one another is fundamental to the biochemistry of all living things. Equilibrium sedimentation is a classic method of biochemistry that provides first-principle thermodynamic information about the molar mass, association energy, association stoichiometry, and thermodynamic nonideality of molecules in solution. It is one of the most rigorous, powerful, and readily adapted methods for characterizing solution interactions. PMID- 18429159 TI - Titration microcalorimetry. AB - Isothermal titration calorimetry (ITC) is perhaps the most rigorous commercially available method for characterizing protein-ligand interactions. In this method, interactions are detected by the intrinsic heat (binding enthalpy) change of the reaction. The technique is applicable to native, unmodified proteins in solution. This is important for proteins that lose or change their functional behavior when chemically modified or attached to a surface. ITC is also useful for evaluating qualitative questions such whether a proposed binding interaction occurs at all, or for quantitatively measuring the concentration of functionally active protein. Finally, if executed with proper control experiments, ITC can be a rich source of thermodynamic information about the molecular binding mechanism. PMID- 18429160 TI - Reduced-scale large-zone analytical gel-filtration chromatography for measurement of protein association equilibria. AB - The proteasome plays a central role in eukaryotic cells since it is responsible for the degradation of specific proteins involved in a large range of cellular processes. Analysis of proteasome mechanisms of action, or in vitro reconstitution, or dissection of the complex biological pathways in which it partakes, requires a reliable source of pure active proteasome. Although the biologically relevant form of the proteasome is usually considered to be the 26S proteasome, this unit describes different methods for purification and study of both 26S and 20S proteasomes from Saccharomyces cerevisiae cells. PMID- 18429161 TI - Size-exclusion chromatography with on-line light scattering. AB - This unit describes the use of size-exclusion chromatography with on-line light scattering, UV absorbance, and refractive index detectors (SEC-LS/UV/RI) to determine: (a) the molecular weight of simple proteins containing no carbohydrates, (b) the molecular weight of glycoproteins, and (c), most importantly, the molecular weight and stoichiometry of protein-protein complexes or protein-carbohydrate complexes. Multiangle light scattering is also discussed. PMID- 18429162 TI - Proteases. AB - The processes of growth and remodeling of cells and tissues in multicellular organisms require the breakdown of old protein molecules, in concert with the synthesis of new ones. For example, many newly-synthesized molecules require proteolytic processing to convert them to biologically active forms. Proteolysis can terminate the activity of a protein--e.g., capsases mediate apoptosis, which is a vital step in the life cycle of the cell. Proteolysis contributes to defense systems too, as the recognition of peptide fragments of foreign proteins triggers the immune response. Proteases are the class of enzymes involved in these important reactions. This unit discusses the general categories of proteases, and sets the stage for addition of overview units on cysteine proteases, aspartic proteases, and metalloproteases, as well as protocol units featuring techniques for analyzing mammalian and yeast proteasomes and protease inhibitors, among other topics. PMID- 18429163 TI - Papain-like cysteine proteases. AB - The name "cysteine protease" refers to the protease's nucleophilic cysteine residue that forms a covalent bond with the carbonyl group of the scissile peptide bond in substrates. The papain-like cysteine proteases, classified as the "C1 family" are the most predominant cysteine proteases. These proteases are found in viruses, plants, primitive parasites, invertebrates, and vertebrates alike. Mammalian papain-like cysteine proteases are also known as cathepsins. This unit discusses cathepsins, and their subcellular and tissue localization, catalytic mechanism, and substrate specificity. Several tables illustrate the properties of the various cathepsins. PMID- 18429164 TI - Overview of pepsin-like aspartic peptidases. AB - The aspartic peptidase family of enzymes has been implicated in a variety of disease states, from stomach ulcers, to breast cancer, and even Alzheimer's Disease. This unit describes the major characteristics of the aspartic peptidases, including mechanism of action, subcellular and tissue localization, and biological substrate specificity. PMID- 18429165 TI - Metalloproteases. AB - Metalloproteases (metallopeptidases) are composed of a diverse group of endopeptidases and exopeptidases. Because metalloproteases play key roles in many normal biological processes, their aberrant activities have been implicated in diseases such as arthritis, cancer, cardiovascular diseases, nephritis, disorders in the central nervous system, fibrosis, and infection, as well as others. This unit provides an overview of metalloproteases, including their classification, catalytic mechanisms, structures, substrate specificities, and common inhibitors. PMID- 18429167 TI - Purification of the eukaryotic 20S proteasome. AB - The 20S proteasome is the catalytic core of the major extralysosomal proteolytic system of the cell. Combination of the 20S proteasome with a complex of regulatory proteins forms the 26S proteasome, which in turn is responsible for the recognition and degradation of ubiquitin-protein conjugates. As described in this unit, the constitutive form of the 20S proteasome can be conveniently purified as a stable and homogeneous preparation from bovine pituitaries. A support protocol details an enzyme assay used in evaluating proteasomal activity. The 20S proteasome is the catalytic core of the major extralysosomal proteolytic system of the cell. PMID- 18429166 TI - Purification and characterization of proteasomes from Saccharomyces cerevisiae. AB - The proteasome plays a central role in eukaryotic cells since it is responsible for the degradation of specific proteins involved in a large range of cellular processes. Analysis of proteasome mechanisms of action, or in vitro reconstitution, or dissection of the complex biological pathways in which it partakes, requires a reliable source of pure active proteasome. Although the biologically relevant form of the proteasome is usually considered to be the 26S proteasome, this unit describes different methods for purification and study of both 26S and 20S proteasomes from Saccharomyces cerevisiae cells. PMID- 18429168 TI - Quantitative amino acid analysis. AB - Colorimetric dye-binding assays are often used to determine protein concentration, however, many proteins do not bind dyes in the same way as the proteins normally used as standards, making quantitation by those techniques difficult. This unit describes the information that can be derived from quantitative amino acid analysis (including precise determination of protein concentration), presents some details on sample preparation, and gives examples of the calculations that are needed. PMID- 18429169 TI - In vitro radiolabeling of peptides and proteins. AB - Radiolabeling of peptides or proteins is often performed to enhance the sensitivity of detection, to quantitate the binding of peptides to other molecules, or for radioimmunoassays. This unit presents a variety of assays for radiolabeling peptides and proteins with (125)I, (131)I, (14)C, and (3)H. PMID- 18429170 TI - Kinetic assay methods. AB - The purpose of this unit is to provide a brief review of issues important in the design of initial-rate assay methods. General aspects of kinetic assay design are discussed, including enzyme and substrate purity, concentration and stability. Also covered are issues such as continuous versus stop-time assay formats, coupled enzyme assays, binding studies, and presentation of initial-rate data. PMID- 18429171 TI - Biotinylation of proteins in solution and on cell surfaces. AB - This unit contains three protocols for biotinylation of isolated proteins: attaching biotin to primary amines (e.g., amino groups of lysyl residues); attaching biotin to sulfhydryls (i.e., thiol groups of cysteinyl residues); and attaching biotin to carbohydrate residues on proteins. As biotinylation of lysyl and cysteinyl residues may alter protein function, modification of protein carbohydrates, which is usually innocuous, may be preferable if intact protein function is required (e.g., for activity assays or affinity purification). Biotinylation of cysteinyl thiols requires that disulfide bonds in isolated proteins be reduced before labeling. Biotinylation of surface proteins on living cells is also described using mild reaction conditions. Finally, this unit includes a brief description of methods for detecting biotinylated proteins. PMID- 18429172 TI - Metabolic labeling with amino acids. AB - Metabolic labeling techniques are used to study biosynthesis, processing, intracellular transport, secretion, degradation, and physical-chemical properties of proteins. This unit focuses on pulse-labeling and pulse-chase experiments done with [(35)S]-methionine, but gives directions for labeling with other radiolabeled amino acids, and also offers guidance for the safe use and handling of 35S-labeled compounds. PMID- 18429174 TI - Selective precipitation of proteins. AB - Selective precipitation of proteins can be used as a bulk method to recover the majority of proteins from a crude lysate, as a selective method to fractionate a subset of proteins from a protein solution, or as a very specific method to recover a single protein of interest from a purification step. This unit describes a number of methods suitable for selective precipitation. In each of the protocols that are outlined, the physical or chemical basis of the precipitation process, the parameters that can be varied for optimization, and the basic steps for developing an optimized precipitation are described. PMID- 18429173 TI - Analysis of selenocysteine-containing proteins. AB - Representatives of three primary life domains--bacteria, archaea, and eukaryotes- possess specific selenium-containing proteins. The majority of naturally occurring selenoproteins contain an amino acid, selenocysteine, that is incorporated into protein in response to the code word UGA. The presence of selenium in natural selenoproteins and in proteins in which this element is introduced by chemical or biological manipulations provides additional opportunities for characterizing structure, function, and mechanism of action. This unit provides an overview of known selenocysteine-containing proteins, examples of targeted incorporation of selenium into proteins, and methods specific for selenoprotein identification and characterization. PMID- 18429175 TI - Production of recombinant proteins in Escherichia coli. AB - E. coli is the expression system of choice and a substantial body of literature has accumulated on the successful expression of foreign genes in this host. Several problems with protein expression in E. coli have been encountered, and many have been ultimately solved. This unit describes methods that have been developed for production of recombinant proteins in E. coli and potential pitfalls that may be encountered. PMID- 18429176 TI - Production of recombinant proteins in mammalian cells. AB - The best strategy for consistent production of larger quantities of pure protein is stable expression. Popular hosts for stable expression are Chinese hamster ovary (CHO) cells, baby hamster kidney (BHK-21) cells, myeloma cells, and the transformed kidney cell line 293. Protocols for stable production in CHO cells are described in this unit. Typical methods for transfection using commercially available plasmid expression vectors are described, along with methods to select for stable expression and methods for amplifying the expression level in the transfected cell. Following this, procedures are presented for efficient cell growth to obtain significant amounts of protein product. Support protocols describe freezing of cells, determination of growth rates, determination of specific productivity of cells, preparing samples for assay, and setting up a 10 day shaker-flask growth curve. PMID- 18429177 TI - Overview of the vaccinia virus expression system. AB - The vaccinia virus expression system differs from others in that transcription occurs in the cytoplasm of mammalian cells rather than in the nucleus. As a vector, vaccinia virus has a number of useful characteristics, including a capacity that permits cloning large fragments of foreign DNA (20+ kbp) with retention of infectivity, a wide host range, a relatively high level of protein synthesis, and "appropriate" transport, secretion, processing, and posttranslational modifications as dictated by the primary structure of the expressed protein and the cell type used. This overview discusses the life cycle of the vaccinia virus along with effects of vaccinia infection. The vaccinia vector expression system is described along with specific steps for expressing genes using these vectors. Important safety considerations are also presented. PMID- 18429178 TI - Preparation of cell cultures and vaccinia virus stocks. AB - This unit describes the maintenance of cell lines used with vaccinia virus, both in monolayer cultures and in suspension. The suspended cell culture is then used in the preparation of vaccinia virus stocks. The preparation of chick embryo fibroblasts (CEF) is also presented for use in the production of the highly attenuated and host range-restricted modified vaccinia virus Ankara (MVA) strain of vaccinia virus. Additionally, support protocols are presented for the titration of standard and MVA vaccinia virus stocks. PMID- 18429179 TI - Generation of recombinant vaccinia viruses. AB - This unit first describes how to infect cells with vaccinia virus and then transfect them with a plasmid-transfer vector to generate a recombinant virus. Methods are also presented for purifying vaccinia virus and for isolating viral DNA, which can be used during transfection. Also presented are selection and screening methods used to isolate recombinant viruses and a method for the amplification of recombinant viruses. Finally, a method for live immunostaining that has been used primarily for detection of recombinant modified vaccinia virus Ankara (MVA) is presented. PMID- 18429180 TI - Characterization of recombinant vaccinia viruses and their products. AB - After a recombinant vaccinia virus is made, its DNA and protein products can be analyzed in several ways. Protocols are provided in this unit for identification of the recombinant virus by PCR (with verification of correct insertion of the DNA by Southern blotting) and by dot-blot hybridization. Also, when antibodies are available, protein expression can be analyzed by immunological methods detailed here such as dot blotting with an antibody, immunoblotting and/or immunoprecipitation. In addition, immunostaining can be used for identification of recombinant plaques as well as for determination of the purity of a recombinant virus stock. All of the protocols in this unit can be used for characterization of modified vaccinia virus Ankara (MVA) recombinant viruses. PMID- 18429181 TI - Gene expression using the vaccinia virus/ T7 RNA polymerase hybrid system. AB - This unit describes a transient cytoplasmic expression system that relies on the synthesis of the bacteriophage T7 RNA polymerase in the cytoplasm of mammalian cells. A gene of interest is inserted into a plasmid such that it comes under the control of the T7 RNA polymerase promoter (p(T7)). Using liposome-mediated transfection, this recombinant plasmid is introduced into the cytoplasm of cells infected with vTF7-3, a recombinant vaccinia virus encoding bacteriophage T7 RNA polymerase. During incubation, the gene of interest is transcribed with high efficiency by T7 RNA polymerase. For large-scale work, protocols are provided for insertion of the p(T7)-regulated gene into a second recombinant vaccinia virus by homologous recombination and subsequent coinfection with vTF7-3 into cells grown in suspension or for direct transfection into OST7-1 cells (a stable cell line that constitutively expresses the T7 RNA polymerase). Expressed protein is then analyzed by pulse-labeling and purified. One new development to this vaccinia virus/T7 RNA polymerase hybrid expression system described here is the VOTE inducible expression system, which eliminates the need to use two recombinant viruses or a special cell line. PMID- 18429182 TI - Choice of cellular protein expression system. AB - Recombinant protein expression has become a standard laboratory tool, and a wide variety of systems and techniques are now in use. Because there are so many systems to choose from, the investigator has to be careful to use the combination that will give the best results for the protein being studied. This overview unit discusses expression and production choices, including post-translational modifications (e.g., glycosylation, acylation, sulfation, and removal of N terminal methionine), in vivo and in vitro folding, and influence of downstream elements on expression. PMID- 18429183 TI - Selection of Escherichia coli expression systems. AB - This unit lists the most useful expression strains of E. coli for fermentation processes. Standard procedures are provided for several expression systems, namely, temperature induction via the p(L) promoter and chemical induction via the trp promoter, lac or tac promoters, and the T7 promoter. These protocols require that the gene encoding the protein of interest has been identified and cloned into an appropriate expression vector using standard molecular biology techniques. Transformation of a suitable host strain (e.g., by electroporation) is also described and is a prerequisite. Protocols for the analysis of plasmid stability and subsequent storage are provided. Support protocols describe how to prepare samples for electrophoresis, how to analyze the solubility of the expressed proteins, and how to make samples of periplasmic extracts and extracellular media (using TCA precipitation). Many of the support protocols are small-scale analysis procedures that are used to guide subsequent purification strategies and determine the suitability of the expression system for further development and scale-up. PMID- 18429184 TI - Fermentation and growth of Escherichia coli for optimal protein production. AB - Large-scale production of recombinant proteins in Escherichia coli requires growth of cells in fermentors. This unit lists E. coli strains appropriate for use in fermentors, and also discusses important characteristics of fermentation equipment. Production of recombinant proteins in batch fermentations is described, as are variations of fermentation systems that enable continuous growth and protein production in high-cell-density, fed-batch cultures and that permit labeling of recombinant proteins with heavy atom derivatives such as selenomethionine or with stable isotopes such as (2)H, (13)C, and (15)N. Production of labeled proteins facilitates structural studies by X-ray crystallography and nuclear magnetic resonance (NMR) spectroscopy. The protocols in this unit are designed for expression systems directing intracellular or periplasmic localization of recombinant proteins; however, in the case of extracellular secretion of the desired protein, the culture medium itself, rather than pelleted cells, would be saved, concentrated, and subjected to purification processes. Fermentation experiments require careful monitoring of cell growth and assurance of preinoculation sterility, both which are described here. PMID- 18429185 TI - Overview of the baculovirus expression system. AB - Baculoviruses have emerged as a popular system for overproducing recombinant proteins in eukaryotic cells. This overview unit describes the baculovirus life cycle and expression system, and also provides information on vectors and protocols for using the baculovirus expression system. PMID- 18429186 TI - Protein expression in the baculovirus system. AB - Insect cell-recombinant baculovirus co-cultures offer a protein production system that complements microbial systems by providing recombinant proteins in soluble form and with most post-translational modifications. Moreover, the large size of the viral genome enables cloning of large segments of DNA and consequent expression of complex protein aggregates. This unit describes methods associated with the large-scale production of recombinant proteins in the baculovirus expression system. A method for large-scale production of viral stocks is described and methods for titration of virus are provided (a plaque assay and an end-point assay). Once viral stocks have been prepared and titered, a protocol for testing the virus in small-scale cultures is provided to determine the kinetics of expression, which allows evaluation of various cell culture and infection conditions aimed at developing optimal levels of protein production (e.g., comparisons of different host cell lines, media, and environmental parameters). Support protocols provide instructions for preparing culture samples for protein analysis by SDS-PAGE and discuss analytical methods for monitoring nutrient levels in cell culture fluids. Once optimal process parameters are identified, protocols describe production of the target protein on a large scale in fermentors using either regular batch production in bioreactors or a fed-batch procedure of production in perfusion cultures. Techniques for harvesting cultures from bioreactors are also provided. PMID- 18429187 TI - Overview of protein expression in Saccharomyces cerevisiae. AB - This overview presents vectors and host strains that are available to direct gene expression in S. cerevisiae, including information on promoters, vector maintenance and copy number, transcription terminators, and selectable markers. Challenges to the expression of foreign proteins are also covered, including attainment of desired production yield, production of protein with appropriate post-translational modifications, conformation and function, and secretion to the extracellular medium. PMID- 18429188 TI - Overview of protein expression in Pichia pastoris. AB - Pichia pastoris is a methylotrophic yeast and can be used as a heterologous expression system. This microorganism is as easy to manipulate as Escherichia coli, but has many of the advantages of eukaryotic expression (e.g., protein processing, folding, and post-translational modifications), and it is faster, easier, and cheaper to use than other eukaryotic expression systems, such as baculovirus or mammalian tissue culture. It also generally yields higher expression levels. This overview discusses important considerations for the use of Pichia pastoris, including strains for expression, expression plasmids, transformation by integration, and post-translational modifications. Examples of expression are given and finally, legal issues regarding patent rights for heterologous protein expression in Pichia pastoris are described. PMID- 18429189 TI - Culture of yeast for the production of heterologous proteins. AB - This unit describes culture of the yeast strains Saccharomyces cerevisiae and Pichia pastoris for the production of foreign proteins. The protocols listed here for S. cerevisiae are for three widely used types of promoter: galactose regulated (GAL1, GAL7, GAL10), glucose-repressible (e.g., ADH2), and constitutive glycolytic (e.g., PGK or GAPDH). Minor variations to each can be made depending on the selection system used. The P. pastoris expression system uses integrating vectors with the methanol-regulated AOX1 promoter and HIS4 selection marker; although transformants are stable, they are generally grown in minimal selective medium. Methods are described for small-scale S. cerevisiae and P. pastoris cultures and also for high-density fermentations with these yeasts. A simple feeding strategy based on calculated feed rates is provided for S. cerevisiae and yields cell densities of 10 to 30 g/liter. In contrast, with P. pastoris, basic fermenter equipment is used to obtain extremely high-density cultures (e.g., 130 g/liter). Finally, a Support Protocol describes small-scale preparation of protein extracts. PMID- 18429190 TI - Overview of protein expression by mammalian cells. AB - This unit reviews the stages involved in protein production in mammalian cells using a stable-expression approach. Choice of cell type is discussed, as is transfection of the host cells, methods for selection and amplification of transformants, and growth of cells at appropriate scale for protein production. Since post-transcriptional modification and intracellular protein transportation are important features of recombinant-protein production in mammalian cells, some description of these mechanisms is included. PMID- 18429191 TI - Overview of protein folding. AB - This overview discusses aspects of protein folding including aggregation, folding pathways, disulfide bonds, stabilization of the native functional state, and approaches for achieving an acceptable rate of protein folding starting with a denatured polypeptide. PMID- 18429192 TI - Folding and purification of insoluble (inclusion body) proteins from Escherichia coli. AB - Heterologous expression of recombinant proteins in E. coli often results in the formation of insoluble and inactive protein aggregates, commonly referred to as inclusion bodies. To obtain the native (i.e., correctly folded) and hence active form of the protein from such aggregates, four steps are usually followed: (1) the cells are lysed and the are aggregates, (2) the cell wall and outer membrane components of the aggregates are removed, (3) the aggregates are solubilized (or extracted) with strong protein denaturants, and (4) the solubilized, denatured proteins are folded with concomitant oxidation of reduced cysteine residues into the correct disulfide bonds to obtain the native protein. This unit features three different approaches to the final step of protein folding and purification. In the first, guanidine HCl is used as the denaturant, after which the solubilized protein is folded (before purification) in an "oxido-shuffling" buffer system to increase the rate of protein oxidation. In the second, acetic acid is used to solubilize the protein which is then partially purified by gel filtration before folding, and then the protein is folded and oxidized by simple dialyzed against water. A Support Protocol is included for rapidly determining the amount of folded protein that contains the correct disulfide linkage pattern. Finally, folding and purification of a fusion protein is described using metal chelate affinity chromatography. PMID- 18429193 TI - Expression and purification of GST fusion proteins. AB - An increasingly common strategy for expressing proteins and large peptides in prokaryotic systems is to express the protein of interest connected to a "tag" that provides the basis for rapid high-affinity purification. This unit describes the expression and purification of fusion proteins containing the 26-kDa glutathione-S-transferase protein as well as methods for cleaving the affinity tag and repurifying the target protein. Advantages of this popular fusion protein system include high protein yields, high-affinity one-step protein purification of the fusion protein, existence of several alternative protease cleavage sites for removing the affinity tag when required, and ease of removal of the cleaved affinity tag. PMID- 18429194 TI - Expression and purification of thioredoxin fusion proteins. AB - This unit describes a gene fusion expression system that uses thioredoxin, the product of the Escherichia coli trxA gene, as the fusion partner. The inherent thermal stability of thioredoxin and its susceptibility to quantitative release from the E. coli cytoplasm by osmotic shock can also be exploited as useful tools for thioredoxin fusion protein purification. In this protocol, a fusion of trxA to any desired gene is constructed and the resulting fusion protein is expressed in an appropriate host strain. Additional protocols describe E. coli cell lysis and fractionation, osmotic release of thioredoxin fusion proteins from the E. coli cytoplasm, and heat treatment to purify some thioredoxin fusion proteins. PMID- 18429195 TI - Overview of the characterization of recombinant proteins. AB - This overview provides guidelines for the characterization of recombinantly expressed proteins (e.g., verifying primary structure and appropriate post translational modifications), along with methodologies for characterizing the proteins according to size, X-ray structure, absorbance, biological activity, and subunit structure. A flow chart presents a suggested path for fully characterizing recombinant protein and involves equipment for HPLC, mass spectrometry, circular dichroism, NMR and fluorescence spectroscopy. Also covered are sources and consequences of contamination in protein solutions. PMID- 18429196 TI - Characterizing recombinant proteins using HPLC gel filtration and mass spectrometry. AB - Recombinant proteins are subject to many forms of heterogeneity, including aggregation, proteolytic degradation, chemical modification, mutation, and incorrect translation. This unit describes methods for the detection and identification of these problems using analytical HPLC gel filtration and MALDI MS. Preliminary characterization of recombinants is necessary before the structure or function of the protein can be investigated. PMID- 18429197 TI - Determining the identity and purity of recombinant proteins by UV absorption spectroscopy. AB - Because each protein (gene product) has a unique amino acid sequence, the particular aromatic amino acid content of each protein results in a unique spectrum in the near-UV (250 to 350 nm) region. The highly specific microenvironment experienced by each aromatic residue in the three-dimensional protein matrix results in fine shifts in a protein's spectrum. This unit provides protocols for the detection and analysis of UV spectra of recombinant proteins and their peptide fragments. The unique UV spectral properties of proteins can in turn be used to assess their purity. This application is inherent in the use of a diode array detector to monitor the effluent from a high-performance liquid chromatography (HPLC) column. A protocol using this technique to assess the purity of recombinant proteins is presented. PMID- 18429198 TI - Determining the identity and structure of recombinant proteins. AB - In this unit peptide mapping protocols with separation of the constituent peptides by high-performance liquid chromatography (HPLC) analysis and by high resolution SDS-PAGE are presented. Peptide mapping is ideally suited for comparative purposes--for example, combined analysis of the recombinant protein and its natural counterpart (or some other well-characterized standard). This unit also outlines the general strategy used to determine the linkage pattern of a monomeric recombinant protein containing two intramolecular disulfide bonds. The approach is an extension of peptide mapping, where the aim is to isolate and characterize peptides containing only a single disulfide bond. A two-dimensional electrophoretic method is also described in which the protein isoelectric point is displayed as a function of pH to yield an electrophoretic titration curve. This method is especially useful for checking for deamidation (e.g., of Asn to Asp) in which additional negative charge is introduced into the modified protein. PMID- 18429199 TI - Transverse urea-gradient gel electrophoresis. AB - Monitoring the cooperative unfolding transition induced when a protein is exposed to elevated temperature or a chemical denaturant is an important strategy for characterizing the conformational properties of a globular protein. This transition may be analyzed quantitatively by a variety of spectroscopic techniques, but a simpler alternative is described in this unit: urea-gradient gel electrophoresis. The pattern produced in the resulting gel can be used to estimate both the free energy change for unfolding and the rate of the unfolding transition. In addition, the technique can help identify either covalent or conformational heterogeneity in a protein sample. Because urea-gradient gel patterns are sensitive to several parameters, including hydrodynamic volume, net charge, and conformational stability, the technique can be particularly useful for comparing two forms of a protein, e.g., a natural form and the product of recombinant bacteria. PMID- 18429200 TI - Analytical ultracentrifugation. AB - Analytical ultracentrifugation is one of the most powerful, though as yet underexploited, techniques available to molecular biology and biochemistry. This overview describes applications for analytical ultracentrifugation along with important considerations relating to experimental design. PMID- 18429201 TI - Light scattering. AB - Light scattering methods can provide information about the native molecular weight, oligomeric composition, and gross conformation of a protein in solution. These methods are particularly well suited for studying large oligomeric systems or glycoproteins and can be used to characterize much larger structures involving protein such as viruses and even bacterial spores. This overview discusses theory and application of both static and dynamic light scattering analysis of protein solutions. PMID- 18429202 TI - Measuring protein thermostability by differential scanning calorimetry. AB - Differential scanning calorimetry (DSC) the method of choice to study the conformational stability of biological macromolecules and proteins in particular. This unit presents step-by-step protocols for DSC, including sample preparation and interpretation of the results in the simplest cases as well as calibration of the apparatus and maintenance of DSC cells. Various general experimental considerations, including possible pitfalls and errors, are also discussed. PMID- 18429203 TI - Overview of conventional chromatography. AB - The key to success in purification of any protein is developing the right separation strategy. This involves selecting and correctly applying a combination of separation techniques, chosen on the basis of information about the target protein and contaminants. There is no single separation technique and no single purification scheme that will allow successful purification of all types of proteins. However, there is a single approach to developing a successful purification strategy. This overview discusses the basic steps that must be taken, including selection of source material, and progression from initial "capture" stage of separation to the final polishing stage. Factors that can affect chromatographic resolution are also covered. PMID- 18429204 TI - Ion-exchange chromatography. AB - This unit outlines the basic steps in planning and carrying out ion-exchange chromatography to separate proteins. Protocols describe both batch adsorption and column chromatography in conjunction with either step- or linear elution gradients. Support protocols describe (1) pilot experiments to determine initial conditions for batch or column chromatography (i.e., pH required for binding, change in pH or salt concentration required for elution, and available capacity of a medium), (2) calculation of the dynamic capacity of an ion-exchange column, (3) methods for producing continuous gradients of pH and salt concentration to elute proteins from ion-exchange columns, (4) regeneration of used ion-exchange media, and (5) storage of ion-exchange media. PMID- 18429205 TI - Gel-filtration chromatography. AB - Three basic protocols are provided in this unit, corresponding to the major applications of gel filtration. One method describes desalting or group separation, in which the target protein and contaminating solutes differ substantially in molecular size. Another is used for protein fractionation, application is in characterizing the molecular dimensions of proteins. A Support Protocol is also provided for calibrating gel filtration columns to be used in estimating molecular size. PMID- 18429206 TI - Hydrophobic-interaction chromatography. AB - This unit discusses the important parameters in designing and optimizing a separation by hydrophobic interaction chromatography (HIC), including preparing the sample and choosing a matrix, column, and buffer. Protocols are provided for packing and testing a column; determining binding and elution conditions; eluting the sample; and cleaning, regenerating, and storing HIC columns. PMID- 18429207 TI - Chromatofocusing. AB - This unit discusses choice of chromatofocusing medium and buffers, determination of the quantity of medium needed, and preparation of CF buffers. A protocol is provided for a typical chromatofocusing experiment, and support protocols describe regeneration of the chromatofocusing column and several methods for separation of the polybuffer from the purified protein. PMID- 18429208 TI - Hydroxylapatite chromatography. AB - Hydroxylapatite (also called hydroxyapatite), a form of calcium phosphate, can be used as a matrix for the chromatography of both proteins and nucleic acids. Protocols are provided for both standard low-pressure chromatography of a protein mixture using a hydroxylapatite column prepared in the laboratory, and an HPLC method, applicable to proteins and nucleic acids, that uses a commercially available column. Alternate protocols describe column chromatography using a step gradient or batch binding and step-gradient elution. PMID- 18429209 TI - HPLC of peptides and proteins. AB - High-performance liquid chromatography (HPLC) is an essential tool for the purification and characterization of biomacromolecules. This unit presents a thorough discussion of the eight types of HPLC currently used, highlighting equipment and start-up procedures, recommendations for running each type of experiment, and theoretical considerations for the separation of peptides and proteins. This is an excellent primer for HPLC users. PMID- 18429210 TI - Lectin affinity chromatography. AB - This unit describes the use of lectins for preparative glycoprotein purification. Con A-Sepharose and WGA-agarose are used for convenience and availability. Instructions are given for a small-scale pilot procedure to test for lectin binding and to determine elution conditions. There are many variations on the basic procedure in the literature, but all use the same principles: bind the protein to immobilized lectin through its sugar chain, wash away unbound protein, and elute bound protein with a simple sugar that resembles the sugar ligand of the bound protein. PMID- 18429211 TI - Dye affinity chromatography. AB - Dye affinity chromatography is a protein purification procedure based on the high affinity of immobilized dyes for the binding sites on many proteins. It is a rapid, inexpensive, and versatile method that is applicable to the purification of crude cellular extracts. This unit presents protocol for the three types of dye affinity chromatography: negative chromatography, positive chromatography, and tandem chromatography. An initial protocol describes a chromatographic procedure in which a small volume of the protein mixture to be purified is applied to a series of miniature columns, each containing a different immobilized dye. Analysis of the flowthrough and bound material allows determination of the optimum dye material for larger-scale purification. An alternate procedure describes a similar initial selection procedure using centrifugation instead of chromatography. A support protocol describes a simple procedure for immobilization of free dyes. PMID- 18429212 TI - Affinity purification of natural ligands. AB - Immobilization of proteins, nucleic acids, and other "bioligands" requires selection of the proper bioligand for the specific application. This decision influences all others: the matrix is chosen and activated by the method that is most appropriate for this specific application and the ligand is coupled under conditions dictated by the activation method and the nature of the ligand (e.g., is it a labile protein or a sturdy enzyme cofactor?). There are many matrices and activation and coupling methods, and new ones are constantly being developed. This unit provides three protocols for activating and coupling proteins and other nucleophilic ligands to beaded agarose gel: cyanogen bromide, p-nitrophenyl chloroformate, and tresyl chloride. PMID- 18429213 TI - Metal-chelate affinity chromatography. AB - Recombinant proteins engineered to have six consecutive histidine residues on either the amino or carboxyl terminus can be purified using a resin containing nickel ions (Ni(2+)) that have been immobilized by covalently attached nitrilotriacetic acid (NTA). This technique, known as metal-chelate affinity chromatography (MCAC), can readily be performed with either native or denatured protein. This unit discusses techniques for creating a fusion protein consisting of the protein of interest with a histidine tail attached. A procedure for expression of histidine-tail fusion proteins and their purification in native form by MCAC is described, and two alternate protocols describe purification of histidine-tail fusion proteins by MCAC under denaturing conditions and their renaturation by either dialysis or solid-phase renaturation. Support protocols are provided for analysis of the purified product and regeneration of the NTA resin. All of these protocols are easily adaptable to any protein expression system. PMID- 18429214 TI - Immunoaffinity chromatography. AB - This unit describes the isolation of soluble or membrane-bound protein antigens from cells or homogenized tissue by immunoaffinity chromatography. This technique involves the elution of a single protein from an immunoaffinity column after prior elution of nonspecifically adsorbed proteins. Specifically, antibodies are coupled to Sepharose (an insoluble, large-pore-size chromatographic matrix). High molecular-weight antigens pass freely into and out of the pores and bind to antibodies covalently bound to the matrix. To elute the bound antigen from the immunoaffinity matrix, the antibody-antigen interaction is destabilized by brief exposure to high- or low-pH buffer. Batch purification of antigens is provided as an alternate procedure that shortens the column loading time. The detergent octyl beta-D-glucoside can be used instead of Triton X-100 for elution. Because octyl beta-D-glucoside has a high critical micelle concentration (CMC), a protocol is provided for its removal by dialysis. The procedure for covalently linking an antibody to Sepharose using the cyanogen bromide activation method is given in a support protocol. PMID- 18429215 TI - Purification of sequence-specific DNA-binding proteins by affinity chromatography. AB - The affinity chromatography procedure described in this unit uses DNA containing specific recognition sites for the desired protein that has been covalently linked to a solid support. Preparation of a DNA affinity resin, including cyanogen bromide (CNBr) activation of the agarose support, is described, and an alternate protocol provides a method to couple DNA to commercially available CNBr activated Sepharose. A method for purification of crude synthetic oligonucleotides by gel electrophoresis prior to preparation of the affinity resin is also provided. A detailed protocol for the actual affinity chromatography procedure is described and a support protocol allows the investigator to determine the appropriate type and quantity of nonspecific competitor DNA that should be used in the procedure and its preparation. Parameters essential to the success of an affinity chromatography experiment are discussed in detail in the Commentary. PMID- 18429216 TI - Purification of DNA-binding proteins using biotin/streptavidin affinity systems. AB - This unit presents purification protocols that exploit the tight and essentially irreversible complex that biotin forms with streptavidin. A DNA fragment containing a high-affinity binding site for the protein of interest is prepared and a molecule of biotinylated nucleotide is incorporated into one of the ends of the DNA fragment. The protein of interest is allowed to bind to the high-affinity recognition site present in the biotinylated fragment. The tetrameric protein streptavidin is then bound to the biotinylated end of the DNA fragment. Next, the protein/biotinylated fragment/streptavidin ternary complex is efficiently removed by adsorption onto a biotin-containing resin. Since streptavidin is multivalent, it is able to serve as a bridge between the biotinylated DNA fragment and the biotin-containing resin. Proteins remaining in the supernatant are washed away under conditions that maximize the stability of the DNA-protein complex. Finally, the protein of interest is eluted from the resin with a high-salt buffer. Both batch and column formats are presented, as is a protocol for the use of streptavidin-agarose. A support protocol describes a mobility shift assay for detecting sequence-specific DNA-binding proteins. PMID- 18429217 TI - Immunoprecipitation. AB - Immunoprecipitation is a technique in which an antigen is isolated by binding to a specific antibody attached to a sedimentable matrix. It is also used to analyze protein fractions separated by other biochemical techniques such as gel filtration or density gradient sedimentation. The source of antigen for immunoprecipitation can be unlabeled cells or tissues, metabolically or intrinsically labeled cells, or in-vitro-translated proteins. This unit describes a wide range of immunoprecipitation techniques, using either suspension or adherent cells lysed by various means (e.g., with and without detergent, using glass beads, etc.). Flow charts and figures give the user a clear-cut explanation of the options for employing the technology. PMID- 18429218 TI - Laboratory safety. AB - This appendix provides protocols for some commonly used disposal and decontamination procedures along with analytical techniques that are used to verify that reagents have been decontaminated. Some of the specific reagents covered are diaminobenzidine, ethidium bromide, cyanogen bromide and chloromethylsilane. With modification, these assays may also be used to determine the concentration of a particular chemical. Precautions are also detailed for routine handling of viable pathogenic microorganisms, as well as all human derived materials, because they may harbor dangerous pathogens such as human immunodeficiency virus (HIV), hepatitis B virus (HBV), cytomegalovirus (CMV), Epstein-Barr virus (EBV), and a host of bacterial pathogens. PMID- 18429219 TI - The polymerase chain reaction. AB - This appendix describes the methodology behind PCR and gives a method for amplifying DNA enzymatically by the PCR and for optimizing this reaction for the sequence and primer set of interest. PMID- 18429220 TI - Purification of glutamate dehydrogenase from liver and brain. AB - Two alternative procedures are described for the purification of the major form of glutamate dehydrogenase (L-glutamate-NAD(P)+ oxidoreductase (deaminating), EC 1.4.1.3: GDH) from ox liver and brain. The first involves affinity chromatography on a column of the allosteric inhibitor GTP bound to Sepharose, whereas the other uses a bifunctional ligand (bis-NAD+) composed of two NAD+ molecules linked together by a spacer arm to precipitate the enzyme in the presence of the substrate analogue glutarate. In both procedures the affinity steps are preceded by ammonium sulfate precipitation and ion exchange chromatography on DEAE cellulose. Procedures for the synthesis of GTP-Sepharose and bis-NAD+ are described and the ancillary procedures, including the assay of GDH activity and the determination of protein concentration, are also presented. PMID- 18429222 TI - Analysis of protein ubiquitination. AB - Attachment of ubiquitin (Ub) to a protein requires a complex of enzymes that recognize the substrate and promote Ub transfer. Sequence motifs present in these enzymes may indicate that other uncharacterized proteins containing these motifs have a biochemical function of Ub-protein ligation, and several in vitro methods are described in this unit for determining if a protein has Ub-transferring activity. They include psmunoblotting of psmunoprecipitated proteins, affinity purification using His-tagged ubiquitin, assaying for auto-ubiquitination of E3, and assaying ubiquitination of a model substrate protein. These methods are suitable for a variety of eukaryotic cells, but techniques are specifically described for use with yeast and mammalian cells. PMID- 18429221 TI - Protein detection in gels using fixation. AB - This unit has been recently updated to include information on preformulated gel stains as well as new protocols for Sypro Ruby and silver staining, and gel imaging methodology. Other previously published protocols are also provided for both rapid and acid-based Coomassie blue staining, and alternate methods for silver staining (i.e., nonammoniacal silver staining, rapid silver staining, and an enhanced-background, two-stage method). PMID- 18429223 TI - De novo peptide sequencing via manual interpretation of MS/MS spectra. AB - Databases of complete genome sequences are rapidly appearing, and a recent trend in interpretation of peptide MS/MS spectra is database matching. A caveat of database matching is that it can only be successful if the database being searched contains the identical amino acid sequence, or one with high homology, to the peptide being investigated. In instances where high-quality MS/MS spectra are obtained for which there is no corresponding database match (e.g., when the peptide of interest is derived from a novel protein from an organism whose genome is not known, has post-translationally modified amino acids, or has multiple amino acid substitutions as compared to the homologous sequence from a different species), de novo (or manual) interpretation of the MS/MS spectrum, as described here, is required. Methods are also provided for verification of the results of the de novo analysis through chemical modification and limited manual Edman degradation of the peptide. PMID- 18429224 TI - Probing protein structure and dynamics by hydrogen exchange-mass spectrometry. AB - Hydrogen exchange rates at peptide backbone amide linkages of proteins are highly sensitive to changes in protein structure and dynamics. Hydrogen exchange-mass spectrometry (HX-MS) can be used to measure hydrogen/deuterium exchange in protein fragments and allows the study of large proteins with high sensitivity and at very low concentrations as well as determination of the intermolecular distribution of deuterium. This unit provides a brief discussion of the interplay between protein dynamics and hydrogen exchange, a detailed prescription for designing and performing typical HX-MS experiments, and common procedures for processing and interpreting HX-MS data. PMID- 18429225 TI - Introduction to atomic force microscopy (AFM) in biology. AB - The atomic force microscope has the unique capability of imaging biological samples with molecular resolution in buffer solution. In addition to providing topographical images of surfaces with nanometer- to angstrom-scale resolution, forces between single molecules and mechanical properties of biological samples can be investigated. Importantly, the measurements are made in buffer solutions, allowing biological samples to stay alive within a physiological-like environment while temporal changes in structure are measured. This overview provides an introduction to AFM on biological systems and describes specific examples of AFM on proteins. The physical principles of the technique and methodological aspects of its practical use and applications are also described. PMID- 18429226 TI - Introduction to peptide synthesis. AB - A number of synthetic peptides are significant commercial or pharmaceutical products, ranging from the dipeptide sugar-substitute aspartame to clinically used hormones, such as oxytocin, adrenocorticotropic hormone, and calcitonin. This unit provides an overview of the field of synthetic peptides and proteins, including their purification. It discusses selecting the solid support and common coupling reagents. Additional information is provided regarding common side reactions and synthesizing modified residues. PMID- 18429227 TI - Synthetic peptides for production of antibodies that recognize intact proteins. AB - Antibodies that recognize intact proteins can be produced through the use of synthetic peptides based on short stretches of the protein sequence, without first having to isolate the protein. The steps to produce an effective antibody include: (1) designing the peptide sequence based on the sequence of the protein; (2) synthesizing the peptide; (3) preparing the immunogen either by coupling the synthetic peptide to a carrier protein or through the use of a multiple antigenic peptide (MAP); (4) immunizing the host animal; (5) assaying antibody titer in the host animal's serum; and (6) obtaining the antiserum and/or isolating the antibody. This unit covers steps 1 and 3. Once the coupling procedure has been performed, it is possible to determine the approximate degree of coupling by amino acid analysis. Also presented are methods for assaying free sulfhydryl content and for reducing disulfide bonds in synthetic peptides. PMID- 18429228 TI - Scintillation proximity assay (SPA) technology to study biomolecular interactions. AB - Scintillation proximity assay (SPA) is a versatile homogeneous technique for radioactive assays which eliminates the need for separation steps. In SPA, scintillant is incorporated into small fluomicrospheres. These microspheres or "beads" are constructed in such a way as to bind specific molecules. If a radioactive molecule is bound to the bead, it is brought into close enough proximity that it can stimulate the scintillant contained within to emit light. Otherwise, the unbound radioactivity is too distant, the energy released is dissipated before reaching the bead, and these disintegrations are not detected. In this unit, the application of SPA technology to measuring protein-protein interactions, Src Homology 2 (SH2) and 3 (SH3) domain binding to specific peptide sequences, and receptor-ligand interactions are described. Three other protocols discuss the application of SPA technology to cell-adhesion-molecule interactions, protein-DNA interactions, and radioimmunoassays. In addition, protocols are given for preparation of SK-N-MC cells and cell membranes. PMID- 18429229 TI - An overview of serine proteases. AB - This unit summarizes the families of serine proteases and their mechanism of catalysis. Methods for assays and determining substrate specificity are briefly described. The mode of action of commonly available inhibitors is also included. PMID- 18429230 TI - Over-expression and purification of active serine proteases and their variants from Escherichia coli inclusion bodies. AB - This unit describes the over-expression and purification of active serine proteases and their variants from E. coli inclusion bodies. The strategy includes the folding and purification of a stable zymogen precursor protein, and its later activation with the appropriate convertase to the less stable but active protease. A test to follow the presence of activity in the samples, together with an active-site titration protocol to determine the number of active sites per mole of total protein are provided. It should be emphasized that although most of the protocols described are applied to a specific example, they are fairly representative of the methods and approaches generally used for laboratory-scale preparation of other recombinant serine proteases. The critical steps and how this template protocol can be adapted for the purification of other serine proteases are described. PMID- 18429231 TI - Assaying proteases in cellular environments. AB - To better study protease activation, regulation, and inhibition physiologically or pharmacologically, it is often advantageous to use cell-based systems. The introduction of the cellular environment poses additional challenges for the assay of the protease of interest. In this unit, various methods for monitoring protease activity in cell-based systems is discussed. The scope of the unit is restricted to mammalian cell system. It is also stressed that specific considerations or requirements for a particular protease or peptidase are often needed in order to optimize the assay. PMID- 18429232 TI - Serpins (serine protease inhibitors). AB - Serpins are a class of proteins involved in the regulation of serine and other types of proteases. In humans, the majority of serpins regulate the functions of proteases involved in the body's response to injury. This includes roles in coagulation, fibrinolysis, inflammation, wound healing, and tissue repair. Serpins have been implicated in various animal and human pathologies by the loss of a functional serpin gene through deletion or mutation, which results in a defect in functional protein. Examples of sestorically called antithrombin III) are first described. Then, protocols to determine the second-order rate constant of AT inhibition of thrombin in the absence and presence of heparin are presented. Also provided is a partial list of other serpins and their purification methods. PMID- 18429233 TI - Caspases. AB - Caspases are a family of cysteine proteases with a strict specificity for aspartate residues involved in inflammatory process and programmed cell death. This overview unit provides basic information on their structure, enzymatic activity, substrate specificity, activation,inhibition and their implication in pathologies. It is intended to be an overview for investigators that are unfamiliar with this family of enzymes but it is also applicable to scientists pursuing research in this field. PMID- 18429234 TI - Use of GFP as a reporter for the analysis of sequence-specific proteases. AB - This unit describes a rapid fluorescent assay for sequence-specific proteases. A recombinant His-tagged substrate-GFP fusion protein containing the sequence specific protease-recognition sequence is used as substrate. Batch metal-chelate chromatography separates uncleaved substrate-GFP fusion protein from GFP released by proteolysis and proteolytic activity is determined by measuring the fluorescence of GFP remaining in solution. PMID- 18429235 TI - Solid-phase profiling of proteins. AB - Protein array technologies refer to any fabrication and use of any arrays containing multiple proteins captured on solid surfaces. This valuable tool is used for high-throughput protein-protein interaction studies, protein marker discovery and other applications. This unit provides basic protocols for biomarker discovery and protein-protein interactions. Different kinds of recently emerged protein array technologies and related detection methods are reviewed. PMID- 18429236 TI - Long-term storage of proteins. AB - This unit provides a summary of some of the issues that researchers face when attempting to store purified proteins. It briefly explains the stresses that induce protein aggregation the major causes for chemical degradation. It also discusses how to use various storage strategies to increase the long-term stability of proteins. When appropriate it points out critical mistakes to avoid. This unit provides a summary of some of the issues that researchers face when attempting to store purified proteins. PMID- 18429237 TI - Overview of the physical state of proteins within cells. AB - To understand how proteins work it is necessary to understand their physical state within the cell. This unit reviews the classification of proteins, how that is related to the hydropathicity of the protein, other factors that affect the heterogeneity of proteins, protein assemblies, methods for altering the solubility of proteins, and limitations of in vitro manipulations of proteins. PMID- 18429238 TI - C-terminal sequence analysis. AB - Carboxy-terminal (C-terminal) sequence analysis is used for direct confirmation of the C-terminal sequence of native and expressed proteins, for detection and characterization of protein processing at the C-terminus, for identification of post-translational proteolytic cleavages, and for obtaining partial sequence information on N-terminally blocked protein samples in order to facilitate design of oligonucleotide probes for gene cloning. This unit describes an automated chemical method and a manual enzymatic (carboxypeptidase digestion) method for determining C-terminal sequence information. Carboxypeptidase digestion requires only a standard amino acid analysis method. PMID- 18429239 TI - Use of protein phosphatase inhibitors. AB - Reversible protein phosphorylation is recognized as a major mechanism regulating the physiology of plant and animal cells. Virtually every biochemical process within eukaryotic cells is controlled by the covalent modification of key regulatory proteins. This in turn dictates the cellular response to a variety of physiological and environmental stimuli; errors in signals transduced by phosphoproteins contribute to many human diseases. Thus, defining protein phosphorylation events, and specifically, the phosphoproteins involved, is crucial for obtaining a better understanding of the physiological events that distinguish normal and diseased states. Protein phosphatase inhibitors are useful when deciphering physiological events regulated by reversible protein phosphorylation but the hormonal stimuli or signaling pathways involved are not known. They are also useful in analyzing the impact of hormones and other physiological stimuli on the function of a specific phosphoprotein. This unit describes protocols for inhibiting the cellular PP1/PP2A activity with okadaic acid, microcystin-LR, and PP2B/calcineurin and a widely utilized strategy for inhibiting protein tyrosine phosphatases. PMID- 18429240 TI - Analytical ultracentrifugation: sedimentation velocity analysis. AB - The analytical ultracentrifuge is a high speed centrifuge with an optical system allowing observation of the concentration of macromolecules as a function of radius and time. In sedimentation velocity experiments,relatively high speeds are used so that a boundary is formed between the solution of sedimenting macromolecule and the buffer in which it is dissolved. Analysis of the rate boundary movement and evolution of its shape can yield information about the molar masses of species present as well as stoichiometries and equilibrium constants for their interactions. This overview discusses crucial issues pertaining to sedimentation velocity experiments, including non-interacting and interacting systems, ideality and non-ideality, and reversible versus kinetically limited equilibrium scenarios. PMID- 18429241 TI - Expression, purification, and characterization of caspases. AB - This unit describes a protocol to obtain milligram amounts of enzymatically active pure recombinant caspases. Specific details for the expression, purification of caspase-3, -6, -7, -8, -9 and -10 are discussed along with strategies to obtain particular forms (e.g., the zymogen) of some of them. PMID- 18429242 TI - Overview of proteome analysis. AB - This unit reviews the new discipline of proteomics, which includes any large scale protein-based systematic analysis of the proteome or defined sub-proteome from a cell, tissue, or entire organism. Proteomics originated in the mid-1990 s due to two key enabling advances, availability of complete genome sequences, and mass spectrometry advances that allowed high sensitivity identifications of proteins. Proteome analyses can be broadly categorized into three types of studies: quantitative protein profile comparisons, analysis of protein-protein interactions, and compositional analysis of simple proteomes or subproteomes such as organelles or large protein complexes. The complexity of different types of proteomes, the merits of targeted versus global proteome studies, and the advantages of alternative separation and analysis technologies are discussed. PMID- 18429243 TI - Protein profiling using two-dimensional difference gel electrophoresis (2-D DIGE). AB - 2D-DIGE relies on pre-electrophoretic labeling of samples with one of three spectrally distinct fluorescent dyes, followed by electrophoresis of all samples in one gel. The dye-labeled samples are then viewed individually by scanning the gel at different wavelengths, which circumvents problems with spot matching between gels. Image analysis programs can then be used to generate volume ratios for each spot, which essentially describe the intensity of a particular spot in each test sample, and thus enable expression differences to be identified and quantified. This unit describes the DIGE procedure in terms of sample preparation from various types of cells, labeling of proteins, and points to consider in the downstream processing of fluorescently labeled samples. PMID- 18429244 TI - Laser capture microdissection for proteome analysis. AB - Laser capture microdissection is being employed increasingly to isolate specific cell types from tissues, thus overcoming problems of experimental interpretation due to tissue heterogeneity of samples. This unit describes protocols which have been optimized to allow laser capture microdissection of tissues with minimal effect on protein integrity, and their subsequent analysis by techniques including 2D-PAGE, immunoblotting and SELDI, and discusses the relative merits of this approach. PMID- 18429245 TI - Use of the gateway system for protein expression in multiple hosts. AB - The Gateway cloning method allows a gene to be cloned and subsequently transferred into any vector by in vitro site-specific recombination. It does not necessarily follow, however, that a gene can be cloned once and expressed in all the available host-vector combinations with uniformly satisfactory results. This is because different organisms have different mechanisms of translating mRNA into protein, and also because choices always have to be made when designing an expression construct, for example, the presence or absence of a stop codon. This unit reviews Gateway cloning, summarizes aspects of protein expression that limit the universality of the use of one clone in many vectors and hosts, and discusses how conflicts between the structure of a Gateway clone of a gene and the rules of protein expression can be minimized or resolved.The Gateway cloning method allows a gene to be cloned and subsequently transferred into any vector by in vitro site specific. PMID- 18429246 TI - Overview of the purification of recombinant proteins produced in Escherichia coli. AB - The updated version of this unit presents an overview of recombinant protein purification with special emphasis on proteins expressed in E. coli. The first section deals with information pertinent to protein purification that can be derived from translation of the cDNA sequence. This is followed by a discussion of common problems associated with bacterial protein expression. A flow chart summarizes approaches for establishing solubility and localization of bacterially produced proteins. Purification strategies for both soluble and insoluble proteins are also reviewed. A section on glycoproteins produced in bacteria in the nonglycosylated state is included to emphasize that, although they may not be useful for in vivo studies, such proteins are well suited for structural studies. Finally, protein handling, scale and aims of purification, and specialized equipment needed for recombinant protein purification and characterization are discussed. The methodologies and approaches described here are essentially suitable for laboratory-scale operations. PMID- 18429247 TI - Rapid screening of E. coli extracts by heteronuclear NMR. AB - Assessing whether a protein or protein complex is amenable to structural analysis is an important component in the structural genomics effort. In particular, if complete sets of structures for entire genomes are to be obtained within a reasonable time frame, high throughput methodologies for all steps along the way have to be developed. These days, cloning and expression systems are highly optimized and a variety of commercially available vectors can be used. However, heterologous proteins or protein domains expressed in bacteria may not be soluble or correctly folded, necessitating intricate solubilization and refolding schemes prior to structural or functional studies. NMR spectroscopy is an important tool for assessing the solubility, stability, and structural integrity of a gene product. It allows efficient evaluation of many variations of polypeptide length and sequence (without time- and labor-intensive purification/refolding procedures) using 1H-15N-HSQC spectroscopy of samples of 15N-labeled proteins directly from crude E. coli extracts. In addition to screening for particular properties of the expressed protein alone, it is also possible to map intermolecular interactions such as ligand binding. In this unit, the basic methodology for bacterial growth, isotope labeling, and spectroscopic evaluation of the protein structure is provided. PMID- 18429248 TI - Determination of disulfide-bond linkages in proteins. AB - The formation of disulfide bonds in proteins is an important post-translational modification that is critical for stabilizing the native structures of proteins. The disulfide linkages in a protein cannot be predicted from its amino acid sequence; therefore, determination of disulfide bond linkages in the protein will provide insights into its three-dimensional structure and contribute to the understanding of its structural-functional relationship. This unit details a series of protocols that have been applied successfully to locate disulfide bonds in proteins. The general strategy involves chemical or proteolytic cleavage of the protein followed by chromatographic separation of the resultant peptides. Disulfide-containing peptides are identified by the alteration of mobility as a consequence of disulfide bond reduction, and are further characterized by mass spectrometry and/or N-terminal sequencing. A partial reduction and alkylation strategy for mapping disulfide linkages in peptides with multiple disulfide bonds is also presented. PMID- 18429249 TI - Enzymatic digestion of proteins in gels for mass spectrometric identification and structural analysis. AB - Enzymatic digestion of proteins is a key technique used in protein identification. By combining the digestion with mass spectrometric detection, proteins at very low femtomole levels, and in some cases subfemtomole levels, can be identified. Typically, one- or two-dimensional SDS-PAGE is used to isolate the proteins of interest, followed by staining with Coomassie blue, digestion compatible silver stain, or Sypro Ruby for detection. Two-dimensional (2-D) fluorescence difference gel electrophoresis (DIGE), which uses Cy3 and Cy5 dyes for detection, allows comparison of two different sample states in order to locate proteins that are up- or down-regulated. In each case, an in-gel digestion, usually tryptic, is used with mass spectrometry to identify these proteins of interest. For large numbers of gel spots, robotic digestion can save time and money. PMID- 18429250 TI - Preparation and application of polyclonal and monoclonal sequence-specific anti phosphoamino acid antibodies. AB - This unit discusses the issues that must be considered in the design, production, and characterization of polyclonal and monoclonal sequence-specific anti phosphoamino acid antibodies. Protocols are provided for generating and purifying such antibodies, and methods are also provided for producing useful polyclonal antibodies in a non-purified form. Support protocols describe coupling of peptides or phosphotyrosine to a solid support for use in affinity chromatography. An example of the generation, purification, and characterization of two sequence-specific anti-phosphopeptide antibodies specific for different sequences of a single phosphoprotein is described. The cross-reactivity of such antibodies, which is a common problem with anti-peptide antibodies, is also discussed. PMID- 18429251 TI - Overview of protein structural and functional folds. AB - This overview provides an illustrated, comprehensive survey of some commonly observed protein-fold families and structural motifs, chosen for their functional significance. It opens with descriptions and definitions of the various elements of protein structure and associated terminology. Following is an introduction into web-based structural bioinformatics that includes surveys of interactive web servers for protein fold or domain annotation, protein-structure databases, protein-structure-classification databases, structural alignments of proteins, and molecular graphics programs available for personal computers. The rest of the overview describes selected families of protein folds in terms of their secondary, tertiary, and quaternary structural arrangements, including ribbon diagram examples, tables of representative structures with references, and brief explanations pointing out their respective biological and functional significance. PMID- 18429252 TI - Crystallization of macromolecules. AB - X-ray crystallography has evolved into a very powerful tool to determine the three-dimensional structure of macromolecules and macromolecular complexes. The major bottleneck in structure determination by X-ray crystallography is the preparation of suitable crystalline samples. This unit outlines steps for the crystallization of a macromolecule, starting with a purified, homogeneous sample. The first protocols describe preparation of the macromolecular sample (i.e., proteins, nucleic acids, macromolecular complexes, and membrane proteins). The preparation and assessment of crystallization trials is then described, along with a protocol for determining whether the crystals obtained are composed of macromolecule or salt. Next, the optimization of crystallization conditions is presented. Finally, protocols that facilitate the growth of larger crystals through seeding are described. PMID- 18429253 TI - Raman spectroscopy of proteins. AB - A protein Raman spectrum comprises discrete bands representing vibrational modes of the peptide backbone and its side chains. The spectral positions, intensities, and polarizations of the Raman bands are sensitive to protein secondary, tertiary, and quaternary structures and to side -chain orientations and local environments. In favorable cases, the Raman spectrum serves as an empirical signature of protein three-dimensional structure, intramolecular dynamics, and intermolecular interactions. Here, the strengths of Raman spectroscopy are illustrated by considering recent applications that address (1) subunit folding and recognition in assembly of the icosahedral capsid of bacteriophage P22, (2) orientations of subunit main chains and side chains in native filamentous viruses, (3) roles of cysteine hydrogen bonding in the folding, assembly, and function of virus structural proteins, and (4) structural determinants of protein/DNA recognition in gene regulatory complexes. Conventional Raman, UV resonance Raman, and polarized Raman techniques are surveyed. PMID- 18429254 TI - Identifying protein interactions by hydroxyl-radical protein footprinting. AB - Hydroxyl-radical protein footprinting is a straightforward and direct method to map protein sites involved in macromolecular interactions. The first step is to radioactively end-label the protein. Using hydroxyl radicals as a peptide backbone cleavage reagent, the protein is then cleaved in the absence and presence of ligand. Cleavage products are separated by high resolution gel electrophoresis. The digital image of the footprinting gel can be subjected to quantitative analysis to identify changes in the sensitivity of the protein to hydroxyl-radical cleavage. Molecular weight markers are electrophoresed on the same gel and hydroxyl-radical cleavage sites assigned by interpolation between the known cleavage sites of the markers. The results are presented in the form of a difference plot that shows regions of the protein that change their susceptibility to cleavage while bound to a ligand. PMID- 18429255 TI - Protein databases on the internet. AB - Protein databases have become a crucial part of modern biology. Huge amounts of data for protein structures, functions, and particularly sequences are being generated. Searching databases is often the first step in the study of a new protein. Comparison between proteins and between protein families in databases provides information about the relationship between proteins within a genome or across different species, and hence offers much more information than can be obtained by studying only an isolated protein. In addition, secondary databases derived from experimental databases are also widely available. These databases reorganize and annotate the data or provide predictions. The use of multiple databases often helps researchers understand the structure and function of proteins. Although some protein databases are widely known, they are far from being fully utilized in the protein science community. This unit provides a starting point for readers to explore the potential of protein databases on the Internet. PMID- 18429256 TI - Expression, purification, and characterization of aspartic endopeptidases: Plasmodium plasmepsins and "short" recombinant human pseudocathepsin. AB - The unit describes a basic protocols utilized to obtain milligram amounts of enzymatically active, pure recombinant Plasmodium plasmepsins and "short" human pseudocathepsin D. Specific details for the expression and purification of Plasmodium falciparum plasmepsin 2 and "short" human pseudocathepsin D in zymogen form are described in this chapter. The plasmepsin 2 protocols are also applicable to Plasmodium vivax, P. ovale, and P. malariae plasmepsins, as well as P. falciparum plasmepsin 4. PMID- 18429257 TI - Zymography of metalloproteinases. AB - Zymography is an electrophoretic technique enabling visualization of the number and approximate size of peptidases in a sample on the basis of their hydrolysis of a protein substrate within the gel. The technique is particularly useful for analyzing the peptidase composition of complex biological samples because visualization depends directly on proteolytic activity. This unit presents a representative zymography protocol for the study of matrix metallopeptidases (MMPs). PMID- 18429258 TI - Monitoring metalloproteinase activity using synthetic fluorogenic substrates. AB - Fluorogenic synthetic substrates are commonly used to monitor the activity of peptidases in vitro. This unit presents a representative protocol that employs (7 methoxycoumarin-4-yl)acetyl-Pro-Leu-Gly-Leu-(3-[2,4-dinitrophenyl]-L-2,3 diaminopropionyl)-Ala-Arg-NH2 (Mca-Pro-Leu-Gly~Leu-Dpa-Ala-Arg-NH2) as a substrate to assay matrix metallopeptidases (MMPs). This substrate was first described for the assay of MMP-1, -2 and -3 and it is now widely used as a general MMP substrate. Protocols are given for both stopped-time assays (suitable for assaying MMP activity in a large number of samples) and continuous assays (commonly used when establishing an assay protocol or investigating kinetic aspects of enzyme behavior). Other fluorogenic peptides and protein substrates, together with non-fluorogenic alternatives, are also discussed. PMID- 18429259 TI - Applications for chemical probes of proteolytic activity. AB - Recent genome sequencing projects have identified new peptidases in multiple organisms, many with unknown functions, suggesting the need for new tools to study these enzymes. This unit outlines selection and use of small-molecule and protein-based probes to covalently modify peptidases in complex cellular environments. These activity-based probes (ABPs) have been designed based on well characterized peptidase inhibitor scaffolds, but make use of new techniques to greatly enhance their utility for studying families of related peptidases. In particular, ABPs can be used to track activity of peptidases in crude cell extracts, intact cells, and in vivo, allowing rapid purification and identification of labeled targets. They can be used with libraries of small molecules to rapidly assess potency and selectivity of compounds in complex, physiologically relevant samples. Probe selection, probe tagging using reporters, labeling of recombinant targets, crude protein extracts, and peptidase targets in cell culture systems, affinity purification of targets, and inhibitor screening using affinity probes are outlined. PMID- 18429260 TI - Preparing protein extracts for quantitative two-dimensional gel comparison. AB - This unit describes basic protocols for efficient and reproducible protein solubilization from a variety of biological samples, including cultured animal cells and tissues, plant cells and tissues, bacteria, nuclei, other subcellular organelles, plasma, serum, and other biological fluids. The optimized extraction process is strongly sample-dependent and cannot be described for every type of sample. Instead, typical protocols are provided as general guidelines and illustrate good starting points for sample-preparation optimization. These solubilization procedures take into account the constraints brought by two dimensional electrophoresis and are thus well suited for proteomic approaches. PMID- 18429261 TI - Isolation of organelles and prefractionation of protein extracts using free-flow electrophoresis. AB - One of the major obstacles in the analysis of proteomes is the extreme complexity of any particular cell or biological fluid. Free-flow electrophoresis (FFE) is a powerful tool for reduction of this complexity, which is a prerequisite for systematic and comprehensive protein analyses. Protocols are provided in this unit for sample fractionation at two different stages: on the protein level by isoelectric focusing FFE fractionation of crude protein mixtures such as whole cell lysates, and on a subcellular level by zone-electrophoretic FFE purification of organelles. PMID- 18429262 TI - Analysis of protein composition using multidimensional chromatography and mass spectrometry. AB - Multidimensional liquid chromatography of peptides produced by protease digestion of complex protein mixtures followed by tandem mass spectrometry can be coupled with automated database searching to identify large numbers of proteins in complex samples. These methods avoid the limitations of gel electrophoresis and in-gel digestions by directly identifying protein mixtures in solution. PMID- 18429263 TI - Quantitative protein profile comparisons using the isotope-coded affinity tag method. AB - Current methods for measuring pairwise changes in protein expression involve differential stable isotopic labeling of proteins or peptides either in vivo or in vitro followed by identification and quantification using a mass spectrometer. In these methods, the mass spectrometer detects two different masses, which correspond to a single protein from two different samples that have been labeled with either a heavy or normal isotope. Changes in protein expression are observed when the identical peptide from each of two biological conditions is identified and a difference is detected in the measurements comparing the peptide labeled with the heavy isotope to the one with a normal isotopic distribution. This approach allows the simultaneous comparison of the expression of many proteins between two different biological states (e.g., yeast grown on galactose versus glucose, or normal versus cancer cells). This unit describes one of these popular methods for quantitative protein profiling using the isotope-coded affinity tag (ICAT) technique. PMID- 18429264 TI - Proteomic analysis using 2-D liquid separations of intact proteins from whole cell lysates. AB - This unit describes procedures for 2-D liquid separations of proteins from whole cell lysates. Protocols for protein isoelectric point (pI) fractionation in the first dimension include the use of liquid isoelectric focusing (IEF) and chromatofocusing. The liquid IEF provides a pI-based fractionation using a batch phase electrophoretic method, while chromatofocusing uses a column-based chromatographic method to generate the pH gradient. Using either method, a second dimension fractionation is provided in the liquid phase using nonporous silica based reversed-phase HPLC (NPS-RP-HPLC) to generate a 2-D liquid map of the protein content of the cell. The eluate of the 2-D liquid fractionation is directly coupled to a mass spectrometer for on-line detection of the intact molecular weights of proteins. As a result, a multidimensional map of protein expression is obtained that characterizes cellular proteins by pI, hydrophobicity, and intact molecular weight. Such expression maps are useful for differential proteomic comparison between different cell samples. PMID- 18429265 TI - Quantitative protein analysis using proteolytic [18O]water labeling. AB - This unit describes the procedure for proteolytic [(18)O]water labeling of peptides in order to quantitate relative protein levels for a comparative proteomic experiment. PMID- 18429266 TI - Spectrophotometric determination of protein concentration. AB - The concentration of a purified protein in solution is most conveniently and accurately measured using absorbance spectroscopy. The absorbance, A, is a linear function of the molar concentration, C, according to the Beer-Lambert law: A = epsilon x l x c, where e is the molar absorption coefficient and l is the cell path length. This unit provides protocols for calculation of epsilon for a folded or unfolded protein, making use of the average epsilon values for the three contributing chromophores in proteins (the side chains of Trp, Tyr, and Cys). A basic protocol describes how to measure the concentration of a protein using the calculated epsilon and the Beer-Lambert law. A sensitive method is provided for measuring the concentration of proteins that contain few if any tryptophan or tyrosine residues, and a simple method is provided for estimating total protein concentration in crude extracts. PMID- 18429267 TI - Overview of cell fractionation. AB - This discussion unit describes the most common methods for cell fractionation which provides the essential ingredients for the increasing number of cell-free assays now being used in test-tube reconstructions of complex cellular events involving intercompartmental interactions. Gel filtration separates on the basis of size, centrifugation separates on the basis of size and density, and electrophoresis separates on the basis of surface charge density. Centrifugation is the most widely used procedure in cell fractionation and is the only approach commonly used to separate crude tissue homogenates (often having quite large volumes) into subfractions as starting material for more refined purification procedures. Therefore, this overview focuses primarily on fractionation of organelles by centrifugation. PMID- 18429268 TI - Purification of organelles from mammalian cells. AB - The protocols in this unit illustrate a range of different procedures that have been used to fractionate tissue homogenates. They emphasize different fractionation techniques that have been used for rat liver, an abundant tissue that has been a favorite of many investigators and has served as the source of many organelle preparations of excellent purity. For selected procedures, other examples have been given using other tissue sources (e.g., glandular tissues that maintain protein storage granules for regulated secretion) or, where particularly favorable, cultured cells. The basis (or goal) of each separation and the merits and limitations of each procedure are summarized to provide a guide for selecting among the various approaches. The large number of protocols include specific details relevant to a particular sample cell type (and in many cases a particular organelle). PMID- 18429269 TI - Subcellular fractionation of tissue culture cells. AB - Cell fractionation techniques include some of the most important and widely used analytical tools in cell and molecular biology, and are essential for the development of cell-free assays that reconstitute complicated cellular processes. In addition to simple gradient systems, this unit discusses the immuno purification of organelles, in particular endosomes. As antigens, purification can be achieved using endogenous or ectopically expressed proteins, provided that appropriate antibodies are available. Alternatively, tagged proteins can be used, when combined with anti-tag antibodies. Now that sequencing of the genomes of several organisms has been completed, biochemical strategies, and in particular fractionation and in vitro transport assays, are more necessary than ever to study the numerous protein and protein complexes that are being discovered. PMID- 18429270 TI - Extraction of proteins from plant tissues. AB - A basic protocol is described for extracting protein from plants. Suggestions are included for overcoming some of the common obstacles encountered (e.g., acidity, phenolics, cell walls). PMID- 18429271 TI - Preparation and extraction of insoluble (inclusion-body) proteins from Escherichia coli. AB - High-level expression of many recombinant proteins in Escherichia coli leads to the formation of highly aggregated protein commonly referred to as inclusion bodies. Inclusion bodies are normally formed in the cytoplasm; alternatively, if a secretion vector is used, they can form in the periplasmic space. Inclusion bodies can be recovered from cell lysates and this unit describes preparation of washed pellets and solubilization of the protein using guanidine x HCl. The extracted protein, which is unfolded, is either directly folded as described in UNIT or further purified by gel filtration in the presence of guanidine x HCl as idescribed here. A support protocol describes the removal of guanidine x HCl from column fractions so they can be monitored by SDS-PAGE. PMID- 18429272 TI - Overview of affinity tags for protein purification. AB - Addition of an affinity tag is a useful method for differentiating recombinant proteins expressed in bacterial and eukaryotic expression systems from the background of total cellular proteins, and for detecting protein-protein interactions. This overview describes the historical basis for the development of affinity tags, affinity tags that are commonly used today, how to choose an appropriate affinity tag for a particular purpose, and several recently developed affinity tag technologies that may prove useful in the near future. PMID- 18429273 TI - Overview of digital electrophoresis analysis. AB - Gel electrophoresis has become a ubiquitous method in molecular biology for separating biomolecules. This prominence is the result of several factors, including the robustness, speed, and potentially high throughput of the technique. The results of this method are traditionally documented using silver halide-based photography followed by manual interpretation. While this remains an excellent method for qualitative documentation of single-gel results, digital capture offers a number of significant advantages when documentation requires quantitation and sophisticated analysis. Digital images of gel electropherograms can be obtained rapidly using an image-capture device, and the images can be easily manipulated using image analysis software. This overview presents reasons for digital documentation and analysis, defines some important key terms for imaging, explains the capture process and reviews the devices used for image capture, and provides an introduction to the software and methods used for one- and two-dimensional digital image analysis. PMID- 18429274 TI - Chemical cleavage of proteins in solution. AB - Described in this unit are five basic protocols that are widely used for specific and efficient chemical cleavage of proteins in solution. Cyanogen bromide (CNBr) cleaves at methionine (Met) residues; BNPS-skatole cleaves at tryptophan (Trp) residues; formic acid cleaves at aspartic acid-proline (Asp-Pro) peptide bonds; hydroxylamine cleaves at asparagine-glycine (Asn-Gly) peptide bonds, and 2-nitro 5-thiocyanobenzoic acid (NTCB) cleaves at cysteine (Cys) residues. Because the above loci are at relatively low abundance in most proteins, digestion with these agents will yield relatively long peptides. PMID- 18429275 TI - Tyrosine O-sulfation. AB - The O-sulfation of tyrosine residues of plasma membrane and secretory proteins that transit through the secretory pathway of eukaryotic cells is a widespread post-translational modification. This enzymatic reaction is catalyzed by trans Golgi-associated tyrosylprotein sulfotransferases, which recognize tyrosine residues located in a specific acidic amino acid sequence. Tyrosine sulfation promotes extracellular protein-protein interactions involved in diverse biological processes, ranging from the receptor binding of regulatory peptides to the interaction of viral envelope proteins with the cell surface. This unit outlines procedures to determine whether a protein of interest contains sulfated tyrosine residues, using methods based on labeling proteins with inorganic [35S] sulfate, alkaline hydrolysis, and one-dimensional thin-layer electrophoresis. PMID- 18429276 TI - Electron microscopy and image processing: an essential tool for structural analysis of macromolecules. AB - Macromolecular electron microscopy (EM) deals with macromolecular complexes and their placement within the cell-linking the molecular and cellular worlds as a bridge between atomic-resolution X-ray crystallographic or NMR studies and lower resolution light microscopy. The amount of specimen required is typically 10(2) to 10(3) times less than for X-ray crystallography or NMR. Electron micrographs of frozen-hydrated specimens portray native structures. Computer averaging yields enhanced images with reduced noise. Three-dimensional reconstructions may be computed from multiple views. Under favorable circumstances, resolutions of 7 to 10 A are achieved. Fitting atomic-resolution coordinates of components into three dimensional density maps gives pseudo-atomic models of a complex's structure and interactions. Time-resolved experiments describe conformational changes. Electron tomography allows reconstruction of pleiomorphic complexes and sub-cellular structures. Electron crystallography has produced near-atomic resolution models of two-dimensional arrays, notably of membrane proteins. PMID- 18429277 TI - Guide for resin and linker selection in solid-phase peptide synthesis. AB - For both novice and experienced practitioners of solid-phase peptide synthesis (SPPS), the vast selection of commercially available linkers and resins has become something of a babel. The purpose of this unit is to clarify the situation, which is best understood by distillation to first principles, through an appreciation of chemical trends and consequences, as well as practical considerations. The most commonly used linkers and resins are presented and described in detail, along with a description of their development and common applications. Key protocols are provided so that the user may prepare appropriate linker-functionalized resins for the majority of peptide synthesis applications. PMID- 18429278 TI - Visualization of protein interactions in living cells using bimolecular fluorescence complementation (BiFC) analysis. AB - Protein interactions integrate stimuli from different signaling pathways and developmental programs. Bimolecular fluorescence complementation (BiFC) analysis has been developed for visualization of protein interactions in living cells. This approach is based on complementation between two fragments of a fluorescent protein when they are brought together by an interaction between proteins fused to the fragments, and it enables visualization of the subcellular locations of protein interactions in the normal cellular environment. It can be used for the analysis of many protein interactions and does not require information about the structures of the interaction partners. A multicolor BiFC approach has been developed for simultaneous visualization of interactions with multiple alternative partners in the same cell, based on complementation between fragments of engineered fluorescent proteins that produce bimolecular fluorescent complexes with distinct spectral characteristics. This enables comparison of subcellular distributions of different protein complexes in the same cell and allows analysis of competition between mutually exclusive interaction partners. PMID- 18429279 TI - Protein charge determination. AB - The most popular current method of determining protein valence entails the calculation of net charge from amino acid sequence/composition. However, the inaccuracy of that approach was recognized long before the advent of the protein data banks and computer programs to facilitate its adoption. Capillary zone electrophoresis affords the simplest and most economical procedure for obtaining a reliable estimate of the net charge of a protein in the buffer system of interest. This unit explains the major pitfalls in the calculation of net charge from protein sequence data. PMID- 18429280 TI - Basic protein sequence analysis. AB - Prediction of molecular function of proteins has become an important task in the genomics era. A wide variety of sequence analysis tools are available to biologists for this task. We have selected one or two primary protocols for tasks such as domain detection, subcellular localization, and motif detection. We also present a strategy for integration of results from different protocols. All the resources needed for these protocols are accessible via publicly available Web servers and databases and require little or no computational expertise. PMID- 18429281 TI - Spectroscopic methods for the determination of protein interactions. AB - This unit provides guidelines on how to use steady-state fluorescence spectroscopy for the quantification of protein-protein interactions. The fluorescence of a protein is characterized by its excitation and emission spectra, quantum yield, and anisotropy. These parameters can change upon interaction with another protein and can be used to measure the extent of complex formation. The source of fluorescence can be an intrinsic fluorophore, such as tryptophan or tyrosine; a covalently attached fluorescent dye; or a fluorescent binding partner, such as a nucleotide or cofactor, that interacts specifically with the complex. Protocols are provided in this unit for determining affinity constants and stoichiometry values for protein-protein interactions using equilibrium titration experiments. In addition, fluorescent labeling of proteins is discussed, and an introduction to data analysis is provided. Most of the topics addressed in this unit can easily be applied to other spectroscopic methods or to the analysis of protein-ligand interactions. PMID- 18429282 TI - Application of amide proton exchange mass spectrometry for the study of protein protein interactions. AB - This protocol describes amide proton exchange experiments that probe for changes in solvent accessibility at protein-protein interfaces. The simplest version of the protocol, termed the "on-exchange" experiment, detects protein-protein interfaces by taking advantage of the fact that solvent deuterium oxide (D2O) molecules are excluded from the surface of a protein to which another protein is bound. A more complete version of the experiment can also be performed in which the rate of surface deuteration is initially measured separately for each of the proteins involved in the interaction, after which the deuterated proteins are allowed to complex and the rate of "off-exchange" (i.e., replacement of surface deuterons by protons from solvent H2O molecules) at the resulting protein-protein interface is measured. This version of the experiment yields additional kinetic information that can help to define the solvent-inaccessible "core" of the interface. PMID- 18429283 TI - Protein profiling by microscale solution isoelectrofocusing (MicroSol-IEF). AB - Sample prefractionation is essential for more comprehensive coverage and reliable detection of low-abundance proteins in complex proteomes. An efficient and reproducible new method for sample prefractionation is microscale solution isoelectrofocusing (MicroSol-IEF), in which samples are separated into chambers defined by membranes of specific pH, yielding well resolved fractions on the basis of isoelectric point (pI). The output seamlessly interfaces with narrow-pH range 2-D gels, enhancing data obtained from protein profiling studies, including quantitative proteome comparisons. This unit presents the MicroSol-IEF method using the ZOOM IEF Fractionator with either commercially available or custom-made pH partition membranes. Alternative configurations are possible for separating samples into different numbers of fractions with various pH ranges and volumes. A detailed method is provided for preparing custom pH membranes. In addition, methods are provided for evaluating the effectiveness of the prefractionation, using 1-D and 2-D gel electrophoresis. Approaches for quantitative protein profiling that incorporate MicroSol-IEF are also discussed. PMID- 18429284 TI - Protein microarrays. AB - With the introduction of DNA microarrays as novel analytical tools, the determination of thousands of binding events in one reaction became possible. The developed technology platforms are not limited to nucleic acids, and, in principle, every ligand-binding assay that works on solid phase can be miniaturized and brought into an array format. This unit explains how protein microarrays can be generated using equipment originally designed for DNA microarrays and how multiplexed assays for the quantification of proteins are set up. A protocol that describes a parallelized system for detecting autoantibodies in human serum is included as an example, and it is shown how existing sandwich immunoassays can be miniaturized and performed in array format. The unit also provides some theoretical background and commentary on the problems associated with this still-novel technology. PMID- 18429285 TI - Human serum and plasma proteomics. AB - This unit focuses on several related methods for profiling and identifying proteins in human serum or plasma. While the methods are specifically described for human serum and plasma, with the possible exception of the major protein depletion steps, they can be readily adapted to serum and plasma from other species as well as to other biological fluids. A very powerful multidimensional protein-profiling method is described that utilizes major protein depletion coupled to two other protein separations, MicroSol IEF fractionation and SDS PAGE, followed by LC-MS/MS. Other protein-profiling strategies include the depletion of abundant proteins followed by analysis either via 2-D gels or using offline Multidimensional Protein Identification Technology (MudPIT) or LC/LC MS/MS. Additional protein profiling methods and strategies are also discussed. PMID- 18429286 TI - Bioinformatics analysis for interactive proteomics. AB - High-throughput protein-protein interaction data are becoming a foundation for new biological discoveries. A major challenge is to manage, analyze, and model these data. In this unit several databases are described that are used to store, query, and visualize protein-protein interaction data. Comparison between experimental techniques reveals that each high-throughput technique has its limitations in detecting certain types of interactions; however, the techniques are generally complementary. In silico prediction methods for protein-protein interactions can expand the scope of experimental data and increase the confidence of certain interactions. Use of protein-protein interaction networks, preferably integrating them with other types of data, allows assignment of cellular functions to novel proteins and derivation of new biological pathways. As demonstrated in this unit, bioinformatics can be used to transform protein protein interaction data from noisy information into knowledge of cellular mechanisms. PMID- 18429287 TI - Misincorporation proton-alkyl exchange (MPAX): engineering cysteine probes into proteins. AB - This unit describes a rapid and efficient method to screen a polypeptide for amino acid residues that contribute to protein-protein interaction interfaces. Cysteine residues are introduced as positional probes in a protein at random by co-expression in bacteria with specific cysteine misincorporator tRNAs. The protein is then purified as an ensemble of polypeptides containing cysteine at low frequency, at different positions in each molecule. The ability of the native protein structure to protect different cysteine residues from chemical modification by iodoacetamide is determined to obtain a protein surface map that reveals candidate surface residues that are likely to be important for protein protein interaction. Cysteine mutants with altered ligand binding can also be selected simultaneously by affinity chromatography. PMID- 18429288 TI - Desalting, concentration, and buffer exchange by dialysis and ultrafiltration. AB - This unit includes a variety of dialysis and ultrafiltration techniques that can be used for desalting, concentration, or buffer exchange. Standard dialysis by diffusion across cellulose tubing is described as a technique for desalting or buffer exchange. Ultrafiltration under pressure can be used either for concentrating protein or, where the sample volume is replenished with a desired buffer, for desalting/buffer exchange (i.e., diafiltration). A variation is ultrafiltration by tangential flow, which serves the same purposes as ultrafiltration under pressure but is better suited to handling large volumes of solution. Yet another variation is an ultrafiltration technique employing centrifugal microconcentrators, which allows concentration of very small volumes of solution. PMID- 18429289 TI - Preparation of soluble proteins from Escherichia coli. AB - Purification of human IL-1beta is used in this unit as an example of the preparation of soluble proteins from E. coli. Bacteria containing IL-1beta are lysed, and IL-1 beta in the resulting supernatant is purified by anion-exchange chromatography, salt precipitation and cation-exchange chromatography, and then concentrated. Finally, the IL-1 beta protein is applied to a gel-filtration column to separate it from remaining higher- and lower-molecular-weight contaminants, the purified protein is stored frozen or is lyophilized. The purification protocol described is typical for a protein that is expressed in fairly high abundance (i.e., >5% total protein) and accumulates in a soluble state. PMID- 18429290 TI - Determining the CD spectrum of a protein. AB - This unit describes the theory behind circular dichroism (CD) and deals with considerations regarding instrumentation and reagents for CD spectrometry. A protocol is provided that outlines the steps in recording a CD spectrum and two support protocols explain the interpretation of near-UV and far-UV CD spectra. PMID- 18429291 TI - Determining the fluorescence spectrum of a protein. AB - Fluorescence spectra of proteins are determined chiefly by the polarity of the environment of the tryptophan and tyrosine residues and by their specific interactions. A thorough consideration of fluorescence spectrometers and their calibration is provided along with important information regarding spectrometer cells, buffers and clarification of samples. Protocols are provided for recording fluorescence spectra and for measuring fluorescence quenching to probe the accessibility of tryptophan residues to small molecules (to yield information about the structural environment of the tryptophan). The technique involves quantifying the decrease in protein fluorescence intensity in the presence of increasing concentrations of quencher, followed by analysis of the data to give details of the interaction of the quencher with the tryptophan residue. Finally, a gives details on how to interpret fluorescence spectra. PMID- 18429292 TI - Expanded-bed adsorption chromatography. AB - Expanded-bed adsorption (EBA) chromatography is a convenient and effective technique for the capture of proteins directly from unclarified crude sample. In EBA chromatography, the settled bed is first expanded by upward flow of equilibration buffer. The crude feed, a mixture of soluble proteins, contaminants, cells, and cell debris, is then passed upward through the expanded bed. Target proteins are captured on the adsorbent, while particulates and contaminants pass through. A change to elution buffer while maintaining upward flow results in desorption of the target protein in expanded-bed mode. Alternatively, if the flow is reversed, the adsorbed particles will quickly settle and the proteins can be desorbed by an elution buffer. The mode used for elution (expanded-bed versus settled-bed) depends on the characteristics of the feed. After elution, the adsorbent is cleaned with a predefined cleaning-in-place (CIP) solution, with cleaning followed by either column regeneration (for further use) or storage. PMID- 18429293 TI - Techniques for mammalian cell tissue culture. AB - This appendix opens with detailed discussions on the latest principles of sterile technique and preparation of culture media. Step-by-step protocols describe trypsinizing and subculturing monolayer cultures, passaging suspension cultures, freezing and thawing cells, counting cells using a hemacytometer, and preparing cells for transport. PMID- 18429294 TI - Endoglycosidase and glycoamidase release of N-linked oligosaccharides. AB - Nearly all proteins entering the lumen of the endoplasmic reticulum (ER) become glycosylated en route to a cellular organelle, the plasma membrane, or the extracellular space. Many glycans can be attached to proteins, but the most common are the N-linked oligosaccharides. These chains are added very soon after a protein enters the ER, but they undergo extensive remodeling (processing), especially in the Golgi. Processing changes the sensitivity of the N-glycan to enzymes that cleave entire sugar chains or individual monosaccharides, which also changes the migration of the protein on SDS gels. These changes can be used to indicate when a protein has passed a particular subcellular location. This unit details some of the methods used to track a protein as it traffics from the ER to the Golgi toward its final location. PMID- 18429295 TI - Determining the structure of oligosaccharides N- and O-linked to glycoproteins. AB - Many proteins involved in biological events are glycosylated. A glycoprotein consists of a mixture of glycosylation variants of a single polypeptide chain, known as glycoforms. It has become clear that a detailed understanding of the roles which glycosylation plays in the biosynthesis, transport, biological function, and degradation of a glycoprotein can only be achieved when the protein and sugar(s) are viewed as an entity. Many glycoproteins can now be modeled by combining glycan sequencing data and oligosaccharide structural information with protein structural data. Pivotal to this approach is sensitive, state-of-the-art oligosaccharide sequencing technology which can give a rapid insight into the glycosylation of a glycoprotein without the need for sophisticated equipment and expertise. This unit gives a detailed introduction into the analysis of glycans, and the many figures will help the user identify which type of experiment needs to be undertaken. Methods for releasing glycans from glycoproteins are followed by protocols for labeling and purifying (by HPLC) the glycans from the rest of the components. Strategies for N- and O-glycan analysis are also included. PMID- 18429296 TI - Determining the structure of glycan moieties by mass spectrometry. AB - Acquisition of mass spectra using fast-atom bombardment (FAB), matrix-assisted laser desorption/ionization (MALDI), and electrospray ionization (ESI) mass spectrometry is summarized in this unit. Molecular weights of the carbohydrates provide information on their composition in terms of isobaric monosaccharide composition, and tables of residue masses are provided to assist these calculations. More detailed structural analysis can be performed by fragmentation; the main fragmentation modes of carbohydrates and their nomenclature are discussed. Fragmentation of negative ions generated by nano electrospray, in particular, provide very informative spectra and give details such as the position of fucose substitution, branching pattern, and the presence or absence of bisecting GlcNAc residues. Practical details are given for the purification of samples prior to mass spectrometric analysis and for recording FAB, MALDI, and ESI spectra. PMID- 18429297 TI - Analysis of protein S-nitrosylation. AB - S-nitrosylation is the binding of an NO group to a cysteine or other thiol. Like phosphorylation, S-nitrosylation is a precisely targeted and rapidly reversible post-translational modification that serves as an on/off switch for protein function during cell signaling. However, unlike phosphorylation, S-nitrosylation of proteins occurs nonenzymatically and is mediated, at least in part, by redox regulated chemical reactions in cells. Alterations in pH, pO(2), cellular reductants, transition metals, and UV light lead to the loss and/or gain of S-NO bonds. Due to the redox-sensitive nature of the modification, analysis of protein S-nitrosylation is technically difficult, since the S-NO bond is easily disrupted during sample preparation. In addition, the level of S-nitrosylated proteins in cells approaches the limit of detection of currently available technology. Despite these technical challenges, several useful methods have been developed recently to measure protein S-nitrosylation in biological samples, and these are described in this unit. PMID- 18429298 TI - Analysis of protein sumoylation. AB - The covalent attachment of small ubiquitin-like modifier (SUMO) proteins to specific lysine residues of target proteins, a process termed sumoylation, is a recently discovered protein modification that plays an important role in regulating many diverse cellular processes. For this reason there is significant interest in identifying new sumoylated proteins and the lysine residue(s) within these target proteins where SUMO attachment occurs. Such knowledge will allow determination of the functional consequences of sumoylation through mutation of the relevant sequences. This unit describes two different experimental approaches for ascertaining specific protein sumoylation: the first is based on immunoprecipitation of the protein of interest followed by SUMO immunoblotting. The second involves incubation of the protein (either an in vitro translation product or a purified recombinant protein) in a reconstituted in vitro sumoylation enzymatic reaction followed by visualization of sumoylated protein as a higher than normal molecular-weight band in SDS-PAGE. PMID- 18429299 TI - Identifying and quantifying sites of protein methylation by heavy methyl SILAC. AB - A new appreciation of protein methylation comes with the recent discovery of demethylases, now placing methylation in the realm of a transient, reversible modification. Classical approaches to study methylation are laborious and involve radioactive, in vitro, enzyme-substrate labeling experiments with purified proteins. Mass spectrometry-based proteomics allows the unbiased analysis of complex protein mixtures and is increasingly applied to the study of post translational modifications. However, it is particularly challenging to study methylation by proteomics because of the number of residues affected and the degree of methylation that can occur. Heavy methyl SILAC is a metabolic labeling strategy that harnesses the cell's machinery to convert a nonradioactive, stable isotope labeled version of methionine into the 'heavy' biological methyl donor S adenosylmethionine. Cells incorporate this 'heavy' methyl group throughout their methylated substrates. This technique increases confidence in identifying and quantifying of sites of protein methylation. PMID- 18429300 TI - Production and use of trimeric isoleucine zipper fusion proteins to study surface receptor ligand interactions. AB - Soluble recombinant forms of surface receptors are commonly used to study the interaction between cell surface receptors and their ligands. By fusing the extracellular domain of the receptor to an isoleucine zipper (ILZ) sequence, it is possible to generate recombinant receptor fusion proteins that form trimers in solution. These ILZ fusion proteins demonstrate several advantages over the commonly used Fc fusion proteins. This unit describes the production and purification of recombinant ILZ fusion proteins. A protocol is also provided for the use of ILZ fusion proteins to study cell surface receptor-ligand interactions on intact cells by flow cytometry. PMID- 18429301 TI - Fluorescence quenching methods to study lipid-protein interactions. AB - This unit describes how fluorescence quenching methods can be used to determine binding constants for phospholipids binding to intrinsic membrane proteins. Reconstitution of a Trp-containing intrinsic membrane protein with bromine containing phospholipids leads to quenching of the Trp fluorescence of the protein; the extent of quenching depends on the strength of binding of the phospholipid to the protein. Protocols are included for the synthesis of bromine containing phospholipids from phospholipids containing carbon-carbon double bonds in their fatty acyl chains and for the reconstitution of membrane proteins into bilayers containing bromine-containing phospholipids. Details are included on data analysis, including equations and software that can be used for fitting the fluorescence quenching data. PMID- 18429302 TI - Overview of Biacore systems and their applications. AB - Surface plasmon resonance (SPR) allows for the investigation of the functional nature of binding interactions and provides detailed kinetic information across a wide range of molecular weights, including small molecules, all without the use of labels. Here the various Biacore instrument platforms and their primary uses, ranging from semi-automated systems designed for simple, flexible basic research to fully automated, high-throughput systems, and systems designed to function in regulated environments, are all highlighted. The available sensor chip surfaces and immobilization techniques are also discussed. Biacore SPR biosensors can be used for a wide variety of assays, including specificity, active concentration measurement, kinetics, and affinity and thermodynamic parameters. Biacore SPR biosensors, which measure real-time analysis of biospecific interactions without the use of labeled molecules, can be used for a wide variety of protein interaction assays. In this unit, examples and recommendations for studying protein interactions with a variety of molecules are provided. This unit also shows how the technology can be used to determine binding specificity, active concentration measurements, and the determination of kinetic and thermodynamic parameters. PMID- 18429303 TI - Using Biacore to measure the binding kinetics of an antibody-antigen interaction. AB - The optical phenomenon of surface plasmon resonance (SPR) used by Biacore systems enables the detection and measurement of protein-protein interactions in real time, without the use of labels. In this unit, the application of Biacore technology to measure a protein-protein interaction is described using an antibody and its antigen as an example. The affinity of the antibody for its antigen is determined by measuring the binding kinetics of the interaction. The protocols are divided into three major steps that are required for measuring binding kinetics using Biacore: (1) surface preparation, (2) assay development, and (3) kinetic analysis. PMID- 18429304 TI - Identifying small-molecule modulators of protein-protein interactions. AB - This unit outlines methods for identifying cyclic peptides that inhibit protein protein interactions. Proteins of interest are cloned into a two-hybrid system engineered to operate in reverse, allowing the disruption of a protein complex to be coupled to cell growth. Cyclic peptide libraries are generated using an intein based plasmid construct, and the cyclized sequence is randomized using a PCR procedure. By transforming plasmid libraries into host cells containing the two hybrid fusions, cyclic peptide inhibitors can be identified by growing the cells under the appropriate selective conditions. A detailed procedure for performing the genetic selection and identifying false positives is provided. Methods for building the two-hybrid protein fusions and optimizing media conditions, as well as an additional protocol for constructing cyclic peptide libraries are also provided. PMID- 18429305 TI - Circular dichroism to study protein interactions. AB - Circular dichroism (CD) is a powerful technique for studying the structures of proteins in solution, as well as structural changes that may occur when proteins bind to ligands. Changes in CD signals accompanying complex formation can be used to determine the strength of the interaction and to provide information about the nature and extent of the conformational changes involved. This unit outlines the type of information available from CD studies and describes how such experiments should be carried out to ensure that reliable information is obtained. PMID- 18429306 TI - Quantitative determination of protein stability and ligand binding by pulse proteolysis. AB - Pulse proteolysis exploits the difference in proteolytic susceptibility between folded and unfolded proteins for facile but quantitative determination of protein stability. The method requires only common biochemistry and molecular biology lab equipment. Pulse proteolysis also can be used to determine the affinity of a ligand to its protein target by monitoring the change in protein stability upon ligand binding. The Basic Protocol describes the detailed procedure for determining protein stability using pulse proteolysis. For pulse proteolysis to be used for determining a protein's stability, the protein should not be digested significantly by pulse proteolysis when it is in the folded conformation. The Support Protocol describes a procedure for determining whether a protein satisfies this requirement. The principles of protein stability determination using denaturant and pulse proteolysis are also discussed. PMID- 18429307 TI - Organelle proteomics. AB - This unit describes strategies for studying the proteomes of organelles, which is one example of targeted proteomics. It relies heavily on previously published units dealing with organelle preparation, protein solubilization, and proteomics techniques. A specific commentary for organelle proteomics is provided. Specific protocols for the isolation of nuclei from various sources (cell cultures, tissues) are also provided. PMID- 18429308 TI - Combinatorial recombination of gene fragments to construct a library of chimeras. AB - Recombination of distantly related and nonrelated genes is difficult using traditional PCR-based techniques, and truncation-based methods result in a large proportion of nonviable sequences due to frame shifts, deletions, and insertions. This unit describes a method for creating libraries of chimeras through combinatorial assembly of gene fragments. It allows the experimenter to recombine genes of any identity and to select the sites where recombination takes place. Combinatorial recombination is achieved by generating gene fragments with specific overhangs, or sticky ends. The overhangs permit the fragments to be ligated in the correct order while allowing independent assortment of blocks with identical overhangs. Genes of any identity can be recombined so long as they share 3 to 5 base pairs of identity at the desired recombination sites. Simple adaptations of the method allow incorporation of specific gene fragments. PMID- 18429309 TI - Wheat germ cell-free expression system for protein production. AB - The Center for Eukaryotic Structural Genomics, in cooperation with Ehime University and CellFree Sciences, has developed a novel wheat germ cell-free technology for the production of eukaryotic proteins. Protein production and purification are robust and scalable for high-throughput applications. The protocols have been used to express and purify proteins from Arabidopsis thaliana, human, mouse, rat and zebra fish. This unit describes expression and purification protocols for both small-scale testing (microgram) and large-scale production (milligram) of N-His6- and N-GST-tagged proteins. The methods described in this unit can be used to produce both unlabeled and labeled proteins required for structure-based determinations by NMR spectroscopy or X-ray crystallography. PMID- 18429310 TI - Protein detection in gels without fixation. AB - A number of alternative methods are described for detecting proteins in polyacrylamide gels that do not require fixation of the protein either prior to staining or in conjunction with staining. The primary advantage of avoiding fixation is that this makes it easier to remove proteins of interest from the gels for subsequent analysis. In general, the sensitivity of protein detection methods that avoid fixation is lower than for detection methods using fixation. For any given method, sensitivity is dependent on the volume of the protein band within the gel; hence, sensitivity is highest for sharp, narrow bands. Techniques described in this unit include protocols for protein detection in gels by SDS precipitation, preparation of contact blots, staining with imidazole-zinc, and use of the fluorescent labels IAEDANS and fluorescamine. Several additional methods, including the use of tryptophan fluorescence, guide strips, and minimal protein staining, are discussed in the Commentary. PMID- 18429311 TI - Crystallization of integral membrane proteins. AB - Over the last 20 years, the use of X-ray crystallography has become a viable technique for the structure determination of integral membrane proteins. However, standard crystallizaton protocols must be modified to account for difficulties involved in handling membrane proteins, which arise primarily from having detergent present. This unit provides protocols that can be used to crystallize a purified membrane protein, including detergent exchange, sample concentration, initial screening using a crystallization robot, and finally, optimization of crystallization conditions to obtain diffraction-quality crystals. These protocols were established for outer membrane proteins, but can be used for inner membrane proteins as well. Advice on alternative protocols, detergent selection, and optimization of crystallization conditions is provided. PMID- 18429312 TI - Overview of solid phase synthesis of "difficult peptide" sequences. AB - Solid-phase peptide synthesis has contributed immeasurably to the understanding of the chemistry and biology of peptides by enabling their ready preparation in small quantities up to the ton scale. The advantages of the technology, including its simplicity, ease of operation, and general efficiency have far outweighed its limitations. However, despite the general effectiveness of the solid phase synthesis methodology, some peptides are resistant to efficient assembly and are known as "difficult peptides." Such sequences can present serious challenges to the peptide researcher and have been the subject of considerable investigation. This phenomenon is described together with modern approaches designed to minimize or overcome this hitherto long-standing and vexing problem. PMID- 18429313 TI - Determination of protein contacts by chemical cross-linking with EDC and mass spectrometry. AB - This unit describes a method for analyzing protein complexes by chemically cross linking closely positioned amino acids. The cross-linked products are isolated by SDS-PAGE and the bands of interest are excised. Proteins in the excised gel piece are digested with trypsin and the resulting peptides recovered. Chemically bonded peptides are distinct from the multitude of unmodified single peptides by the presence of two carboxy-terminal ends. The protocol also describes incorporation of (18)O at the carboxy-terminal ends and purification and preparation of the peptide mixture for mass spectroscopy. PMID- 18429314 TI - Detection of protein-protein interactions by coprecipitation. AB - Co-precipitation of proteins from whole cell extracts is a valuable approach to test for physical interactions between proteins of interest. It may be the method of choice, or may be used in conjunction with other procedures to detect protein protein interactions, such as two-hybrid analyis and co-purification schemes, and tests of physical associations using purified proteins. PMID- 18429315 TI - Minimotif miner: a computational tool to investigate protein function, disease, and genetic diversity. AB - The Minimotif Miner Web site contains information on several hundred short functional motifs in a single database, and allows the user to search protein queries for the presence of these motifs. Scoring based on evolutionary conservation, protein surface prediction, and motif frequency can be used in conjunction with other motif programs and the known biology of the query to reduce false-positive predictions and select short motifs for experimental pursuit. PMID- 18429316 TI - Protein structure analysis online. AB - Computational biology/chemistry tools are used in most areas of life/health science research. These methods are continually being developed and their use can present difficulties for both experienced and novice investigators. To facilitate the use of these applications, many packages have been implemented online during these last 5 years. This unit focuses on online computational methods with a special emphasis on structural refinement/atomic simulations, protein electrostatic calculations, searches for functional sites, searches for druggable pockets, protein docking and small molecule docking, and prediction of potential impact of amino acid variations on the structure and function of the protein molecules. PMID- 18429317 TI - Comparative protein structure modeling using MODELLER. AB - Functional characterization of a protein sequence is a common goal in biology, and is usually facilitated by having an accurate three-dimensional (3-D) structure of the studied protein. In the absence of an experimentally determined structure, comparative or homology modeling can sometimes provide a useful 3-D model for a protein that is related to at least one known protein structure. Comparative modeling predicts the 3-D structure of a given protein sequence (target) based primarily on its alignment to one or more proteins of known structure (templates). The prediction process consists of fold assignment, target template alignment, model building, and model evaluation. This unit describes how to calculate comparative models using the program MODELLER and discusses all four steps of comparative modeling, frequently observed errors, and some applications. Modeling lactate dehydrogenase from Trichomonas vaginalis (TvLDH) is described as an example. The download and installation of the MODELLER software is also described. PMID- 18429318 TI - Proteomic identification of cellular protease substrates using isobaric tags for relative and absolute quantification (iTRAQ). AB - Identification of protease substrates is essential to identify and understand the functional consequences of normal and dysregulated proteolysis in disease on the proteome. Isobaric tags for relative and absolute quantification (iTRAQ) can be used to identify novel protease substrates in the cellular context. An amine targeted iTRAQ tag labels tryptic peptides generated from the proteins and protease cleavage products of secreted proteins, as well as protein domains shed from the cell membrane or pericellular matrix of protease-transfected cells that have accumulated in conditioned medium; a second iTRAQ tag is used for control cells. MS/MS fragmentation enables sequencing of the pooled pairs of differently labeled but identical peptides and generates a low mass signature ion peak unique for each label. This signature ion peak identifies the peptides originating from the protease-transfected or control cells; comparison of the peak areas enables relative quantitation of the peptide between the samples. PMID- 18429319 TI - The CLIP-CHIP oligonucleotide microarray: dedicated array for analysis of all protease, nonproteolytic homolog, and inhibitor gene transcripts in human and mouse. AB - The CLIP-CHIP oligonucleotide microarray allows the analysis of mRNA transcript levels in a tissue sample for all proteases, nonproteolytic homologs, and protease inhibitors of the human and mouse genome. In the protocol presented in this unit, total RNA is extracted from a tissue, and the resulting mRNA is reverse transcribed into cDNA and dsDNA and then amplified in an in vitro transcription reaction. The amplified antisense RNA is labeled with a fluorescent dye and hybridized to the CLIP-CHIP, which contains unique oligonucleotides that are specifically designed for the protease, nonproteolytic homologs, protease inhibitors, and control samples. After hybridization, the fluorescence intensity of each spot is measured, thus identifying mRNA transcripts that are expressed and allowing basic quantification of expressed transcripts. PMID- 18429320 TI - Purification and characterization of gingipains. AB - Gingipains are cysteine proteases produced in large quantities by Porphyromonas gingivalis which together constitute important virulence factors in the pathogenesis of periodontal disease by that organism. Described is this unit is an efficient procedure for the purification of gingipains from the growth medium of P. gingivalis strain HG66, along with detailed protocols for growth of the organism and basic characterization of the purified proteases. The purification procedure consists of acetone precipitation followed by gel filtration to separate high-molecular-mass gingipains (Kgp and HRgpA) from RgpB. Kgp and HRgpA are further separated on Arg-Sepharose by the virtue of differential elution from the affinity matrix with lysine (Kgp) and arginine (HRgpA) eluant. Obtained from these procedures, the gingipains are stable and can be stored at -80 degrees C for years without loss of activity. PMID- 18429321 TI - Microplate assay for cathepsin detection in viable cells using derivatives of 4 methoxy-beta-naphthylamide. AB - This unit describes an assay for the direct and selective detection of the four cathepsins B, H, K, and L in adherently growing cells. Cells are incubated with substrates that are peptidic derivatives of 4-methoxy-beta-naphthylamine partially selective for each cathepsin, together with 5-nitrosalicylaldehyde. The protease reaction is performed in microtiter plates and the fluorescent hydrolysis products are detected using a plate reader. The selectivity of detection is enhanced by parallel assays containing inhibitors that are also partially selective for each of the cathepsins. Individual cathepsin activities can then be calculated by the difference between the uninhibited and the inhibited assays. Detection of cathepsin H activity differs from the other assays in that other nonlysosomal aminopeptidases are inhibited by bestatin. The most common application of these assays is to compare directly cells treated with different substances, e.g., pharmaceutically interesting cathepsin inhibitors. PMID- 18429322 TI - Glycoproteomics using chemical immobilization. AB - Protein glycosylation is prevalent in proteins destined for extracellular environments, e.g., transmembrane proteins, cell surface proteins, and secreted proteins in tissues and body fluids. These also are the proteins that are most easily accessible for diagnostic and therapeutic purposes. This unit describes methods for solid-phase extraction of glycopeptides and subsequent identification of glycopeptides as well as glycosylation sites. The extraction is based on the conjugation of glycopeptides to a solid support, using hydrazide chemistry, and the specific release of formerly glycosylated peptides. The recovered peptides are then identified by tandem mass spectrometry. The methods are applied to the analysis of proteins from cells, body fluids, and tissues. PMID- 18429323 TI - Phosphoproteomics. AB - Protein phosphorylation is one of the most important mechanisms of regulating protein function in cells, and it plays an important role in controlling diverse biological processes, including cellular proliferation, migration, and metabolism. The term "phosphoproteome" refers to the complement of proteins that undergoes phosphorylation, the extent of their phosphorylation status at the level of individual residues, as well as the dynamics of the phosphorylation events in response to various stimuli. This unit provides methods for enrichment of phosphorylated proteins and peptides using anti-phosphotyrosine antibodies or titanium dioxide, respectively. Support protocols are provided for two detergent free cell lysis methods, fractionation of proteins prior to enrichment, and use of stable isotope labeling by amino acids in cell culture (SILAC) method for studying dynamics of phosphorylation events. PMID- 18429324 TI - Overview of tandem mass spectrometry (MS/MS) database search algorithms. AB - Mass spectrometry-based methods for the identification of proteins are fundamental platform technologies for proteomics. One comprehensive approach is to subject trypsinized peptides to tandem mass spectrometry (MS/MS) to obtain detailed structural information. Different strategies are available for interpreting MS/MS data and hence deducing the amino acid sequence of the peptides. The most common method is to use a search algorithm to identify peptides by correlating experimental and theoretical MS/MS data (the latter being generated from possible peptides in the protein sequence database). Identified peptides are collated and protein entries from the sequence database inferred. This unit focuses on the most widely used tandem MS peptide identification search algorithms (commercial and open source), their availability, ease of use, strengths, speed and scoring, as well as their relative sensitivity and specificity. PMID- 18429325 TI - Incorporation of isotopically enriched amino acids. AB - The incorporation of isotope labels into proteins is extremely useful for the application of nuclear magnetic resonance (NMR), X-ray or neutron-diffraction crystallography, and mass spectrometry (MS) methodologies to investigate the structure and dynamics of proteins. This unit presents methods for incorporating isotopic labels into proteins via expression in E. coli and baculovirus transfected Sf9 insect cells or through cell-free means. The unit also presents methods for introducing isotopic labels by chemical means into synthetic peptides by solid phase peptide synthesis or into isolated proteins by chemical modification of labile protein groups. PMID- 18429326 TI - Assays for determination of protein concentration. AB - Biochemical analysis of proteins relies on accurate quantitation of protein concentration. This unit describes how to perform commonly used protein assays, e.g., Lowry, Bradford, BCA, and UV spectroscopic protein assays. The primary focus of the unit is assay selection, emphasizing sample and buffer compatibility. Protein assay standard curves and data processing fundamentals are discussed in detail. This unit also details high-throughput adaptations of the commonly used protein assays, and also contains a protocol for BCA assay of total protein in SDS-PAGE sample buffer that is used for equal loading of SDS-PAGE gels, which is reliable, inexpensive, and quick. PMID- 18429327 TI - Overview on the expression of toxic gene products in Escherichia coli. AB - Despite the development of various nonbacterial expression systems, Escherichia coli (E. coli) remains the host of choice for recombinant protein expression. Its culture is simple, fast, inexpensive, and highly efficient (tens of milligrams of pure proteins are typically obtained within 48 hours using as little as 1 liter of culture). Unfortunately, many toxic genes (from various organisms) severely interfere with the physiology of E. coli. As a result, expression yields are dramatically diminished, and sometimes abolished. In fact, some genes are so toxic that E. coli cannot maintain their expression vector during the growth phase (the phase during which recombinant gene expression is presumably repressed). Therefore, modified expression vectors, modified E. coli strains, and appropriate cultivation protocols are needed. This overview discusses several special strategies successfully used to express toxic genes in E. coli. PMID- 18429328 TI - Automated large-scale purification of a recombinant g-protein-coupled neurotensin receptor. AB - Structure determination of G-protein-coupled receptors and other applications, such as nuclear magnetic resonance (NMR) studies, require milligram quantities of purified, functional receptor protein on a regular basis. This unit presents a step-by-step procedure for the automated two-column purification at the 10 milligram scale of a G protein-coupled receptor for neurotensin, expressed in functional form in Escherichia coli. PMID- 18429329 TI - Fabrication of 3-D gel microarrays directly with raw polymerase chain reaction products by heat-directed polymerization. AB - In this study, a new method for fabricating 3-D polyacrylamide gel microarrays was presented. In the method, a 3-D gel microarray was successfully fabricated by directly heating the slides with arrayed prepolymer at 80 degrees C for 2 min. The prepolymer on the microarray only contained 40% v/v raw/unpurified PCR products, 1.4% ammonium persulfate (APS), 6% acrylamide, and 20% glycerol and the balance H2O but no TEMED. The heat could induce the free radicals from APS to initiate polymerization of the 3-D gel and allow the incorporation of raw PCR products into the 3-D gel. The immobilized DNA on the gel microarray had the specificity of hybridization and the ability to distinguish base mismatch. It has been demonstrated that SNP genotyping of both the 194072 loci and the 252944 loci in the beta2 subunit gene of gamma-aminobutyric acid A receptor (GABRB2) gene was successfully performed by using the 3-D gel microarray. This method is efficient, simple, rapid, and easy to control in preparing a 3-D gel microarray, which would be widely used for various purposes in laboratory research and clinic application. PMID- 18429330 TI - Exonuclease activity of proofreading DNA polymerases is at the origin of artifacts in molecular profiling studies. AB - CE fingerprint methods are commonly used in microbial ecology. We have previously noticed that the position and number of peaks in CE-SSCP (single-strand conformation polymorphism) profiles depend on the DNA polymerase used in PCR [1]. Here, we studied the fragments produced by Taq polymerase as well as four commercially available proofreading polymerases, using the V3 region of the Escherichia coli rss gene as a marker. PCR products rendered multiple peaks in denaturing CE; Taq polymerase was observed to produce the longest fragments. Incubation of the fragments with T4 DNA polymerase indicated that the 3'-ends of the proofreading polymerase amplicons were recessed, while the Taq amplicon was partially +A tailed. Treatment of the PCR product with proofreading DNA polymerase rendered trimmed fragments. This was due to the 3'-5' exonuclease activity of these enzymes, which is essential for proofreading. The nuclease activity was reduced by increasing the concentration of dNTP. The Platinum Pfx DNA polymerase generated very few artifacts and could produce 85% of blunted PCR products. Nevertheless, despite the higher error rate, we recommend the use of Taq polymerase rather than proofreading in the framework for molecular fingerprint studies. They are more cost-effective and therefore ideally suited for high-throughput analysis; the +A tail artifact rate can be controlled by modifying the PCR primers and the reaction conditions. PMID- 18429331 TI - Waiting for HAPO. AB - The recent global increase in gestational diabetes has paralleled the increased prevalence of obesity and type 2 diabetes. The Hyperglycemia and Adverse Pregnancy Outcome (HAPO) study is an attempt to establish gestational diabetes diagnostic criteria as well as to clarify the accurate glucose threshold for the relationship between maternal hyperglycaemia and adverse perinatal outcomes. Although five international workshops have convened to address the importance of gestational diabetes, a resolution towards a general consensus for diagnosis and clinical management of gestational diabetes has not been achieved. Such a resolution may never appear, much like the outcome in the play Waiting for Godot, a character who in the end never arrives, and may not even exist. The accompanying article by Mathiesen and Vaz highlights the fetal, neonatal and maternal risks that accompany inadequate glycaemic control during pregnancies complicated by diabetes, even in the presence of only mild maternal hyperglycaemia. Diet, exercise and an optimised treatment regimen based on regular pre- and postprandial monitoring of blood glucose are essential throughout pregnancy. Pivotal to this goal is the recognition that insulin requirements in pregnancy are distinct from those of the prepregnancy state, and that these requirements change throughout gestation, labour and lactation. PMID- 18429332 TI - Insulin treatment in diabetic pregnancy. PMID- 18429333 TI - Epidemic. HIV/AIDS cases among inmates fell for 6th consecutive year in 2005. PMID- 18429334 TI - Blind, HIV-positive woman denied SSI benefits. PMID- 18429335 TI - Dismissal denied prison officials in hostile-workplace claim. PMID- 18429337 TI - HIV-related defamation claims dismissed against broadcasters. PMID- 18429336 TI - Claim over disclosure of inmate's serostatus dismissed. PMID- 18429338 TI - Abstracts of the 4th Biennial Conference of the International Society of Affective Disorders (ISAD), 14-17 March 2008, Cape Town, South Africa. PMID- 18429339 TI - Abstracts of the 81st Annual Meeting of the Japanese Pharmacological Society, March 17-19, 2008, Yokohama, Japan. PMID- 18429341 TI - Abstracts of the 13th World Congress of Gynecological Endocrinology, February 28 March 2, 2008, Florence, Italy. PMID- 18429340 TI - [Abstracts of the 32nd Francophone Meeting of Digestive Pathology, 8-12 March 2008, Paris, France]. PMID- 18429342 TI - Abstracts of the ECNP (European College of Neuropsychopharmacology) Workshop on Neuropsychopharmacology for Young Scientists in Europe, 6-9 March 2008, Nice, France. PMID- 18429343 TI - Health crisis amid the Maoist insurgency in India. PMID- 18429344 TI - Can you pay people to be healthy? PMID- 18429345 TI - In Memoriam: Niels L. Low, MD (1916-2007). PMID- 18429346 TI - Literary healing. PMID- 18429347 TI - Pten against premature ovarian failure; the importance of the maternal nucleolus. In the spotlight. PMID- 18429348 TI - Acquisition of anal human papillomavirus (HPV) infection in women: the Hawaii HPV Cohort study. AB - BACKGROUND: The majority of anal cancer is associated with human papillomavirus (HPV) infection, yet little is known about women's risk of acquisition of anal HPV infection. METHODS: Risk factors for the acquisition of anal HPV infection were examined in a longitudinal cohort study of 431 women, via repeated measurement of HPV DNA. RESULTS: Seventy percent of women were positive for anal HPV infection at one or more clinic visits from baseline through a follow-up period that averaged 1.3 years. The incidence of a high-risk (HR) infection was 19.5 (95% confidence interval [CI], 16.0-23.6) per 1000 woman-months. The most common incident HR HPV types were HPV-53, -52 and -16. The presence of an HR anal HPV infection at baseline increased the risk of an incident anal infection by 65%. Baseline HR cervical HPV infection also predicted the acquisition of an HR anal HPV infection (odds ratio, 1.81 [95% CI, 1.09-3.02]). Nonviral risk factors for acquisition of HR HPV infection included younger age, lower socioeconomic status, greater lifetime number of sexual partners, past use of hormones, and condom use. CONCLUSIONS: The results of this study suggest that women's risk of anal HPV infection is as common as their risk of cervical HPV infection. PMID- 18429349 TI - Inoue Stent-Graft Implantation for Thoracoabdominal Aortic Aneurysm Involving the Visceral Arteries. AB - Purpose. To assess the efficacy of the Inoue stent-graft placement for thoracoabdominal aortic aneurysm (TAAA).Methods. Patients with TAAA underwent Inoue stent-graft placement with single branched stent-graft in 4 patients,straight graft in 3 patients and double branched stent-graft in 1 patient. Half the patients required additional open surgical revascularizations of involved visceral arteries (Hybrid procedures).Results. Stent-grafts were deployed successfully in all patients. One patient with Hybrid procedure developed major complications,required haemodialysis and died in hospital. In another patient the post-operative CT scan demonstrated a type I endoleak, but this had resolved by 3 months.Conclusion. Inoue stent-grafting for TAAA with or without adjunctive open surgical revascularization is feasible. PMID- 18429351 TI - Sublingual immunotherapy for hazelnut food allergy: a follow-up study. PMID- 18429352 TI - [The passing of a concepts agitator]. PMID- 18429353 TI - Oncology in Belgium: a global approach where research and clinic are joining. PMID- 18429354 TI - [Cancer research in the French speaking community in Belgium]. AB - The author examines the status of cancer research in the French speaking community in Belgium. PMID- 18429355 TI - E-prescribing. PMID- 18429356 TI - Cancer genomics and proteomics. Methods and Protocols. Preface. PMID- 18429357 TI - Dietary approaches to manage body weight. AB - It has been established in the Canadian Community Health Survey that 23% of Canadians are obese while 36% are over weight. The fact that obesity is associated with an increased risk of developing cardiovascular disease, type 2 diabetes and other chronic diseases explains why many efforts have been put forward to develop tools that can be helpful for achieving body weight loss. Nutrition intervention is at the cornerstone of obesity treatment. In order to achieve body weight loss, energy intake must be lower than energy expenditure. This mathematical equation is rather simple and can falsely suggest that achieving long-term weight loss is simple and easy. However, many studies have demonstrated the poor long-term success of weight loss interventions. Biological as well as behavioural and environmental factors can all contribute to body weight regain after weight loss. This paper will discuss some nutritional strategies that can be helpful to induce a reduction in energy intake. Namely, the impact of reducing energy density, reducing portion size and improving the recognition of hunger and satiety signals and diet modification will be discussed. PMID- 18429358 TI - The time bomb of Aboriginal health: can the fuse get any shorter? PMID- 18429359 TI - RE: S Mahmoud, LM Beauchesne, DR Davis, C Glover. Acute reversible left ventricular dysfunction secondary to alcohol. Can J Cardiol 2007;23(6):475-477. PMID- 18429360 TI - Ethnic masculinities in Australian boys' schools: Scots and Irish secondary schools in late nineteenth-century Australia. PMID- 18429361 TI - Foodborne disease trends and reports. PMID- 18429363 TI - Hope and suffering at the Devon county pauper lunatic asylum at Exminster, 1845 1914. PMID- 18429364 TI - [Guidelines for the health surveillance of subjects exposed to carcinogenic and mutagenic agents at the workplace. First review]. PMID- 18429365 TI - The value of peer review: the hard workers behind the scenes. PMID- 18429366 TI - The role of the nurse planner. AB - Accreditation criteria require that at least one designated nurse planner have responsibility for all aspects of an accredited provider unit's functions. This column explores those responsibilities in relation to each of the accreditation criteria. PMID- 18429367 TI - Crohn's disease and ulcerative colitis. AB - This column discusses the prevalence, symptoms, and treatment of Crohn's disease and ulcerative colitis and the role nurses play for these patients. PMID- 18429368 TI - Increasing competency in the care of homeless patients. AB - Nurses play a critical role in helping homeless patients make the transition from revolving door hospitalizations or emergency department visits to ongoing care through an outpatient clinic. This column focuses on increasing competency in the care of homeless patients. The next column will focus on a different type of transition-preparing hospitalized patients for discharge and referral to home health care. PMID- 18429369 TI - Helping faculty enhance scholarship. AB - Nurse educators face a myriad of challenges (e.g., changing student populations, increased demand for the use of technology, faculty shortages, and facilitating the development of self-confidence and competence in students) as they endeavor to enhance scholarship and quality in nursing education. Scholarship encompasses four separate but integrated elements (i.e., discovery, integration, application, and teaching) that need to be instilled in nursing students to prepare them for diverse roles in the profession of nursing. Implications for nurse educators relate to creating curricula that support scholarship, technological and interprofessional opportunities, and strategies for socializing students into scholarship. PMID- 18429370 TI - Nurses reclaiming ownership of their practice: implementation of an evidence based practice model and process. AB - This article describes a new model and process to implement evidence-based practice. This model builds on concepts from the Iowa Model of Evidence-Based Practice, the Stetler model, and Rosswurm and Larrabee's model. The new model focuses on the centrality and involvement of staff nurses in making evidence based practice clinical changes. Two figures illustrate the model and the implementation process. A detailed case study based on the model is included. Barriers identified in the literature review are addressed in the case study. Implementation of this model creates opportunities for staff nurses to recognize ownership of their practice and their role in changing the practice setting to a culture of evidence-based practice. PMID- 18429371 TI - Nursing grand rounds as a medium for the continuing education of nurses. AB - Grand rounds are an educational technique used by the medical profession, yet only sporadically used in nursing. When grand round are employed, knowledge can be disseminated on a consistent basis. No literature was found detailing the effectiveness and value of such programs. This article describes a small pilot study that assessed the effect on knowledge and the perceived value of a nursing grand rounds (NGR) format. To measure these two concepts, those who attended the inaugural NGR presentation were given a posttest and an assessment survey 1 year after the presentation. The posttest focused on key concepts presented, and the survey rated perceived value and knowledge on a scale from 1 (strongly disagree) to 5 (strongly agree). Results demonstrated positive knowledge acquisition and a high perceived value. This assessment review lends support to the use of institutional NGR as a means of continuing education and professional advancement. PMID- 18429372 TI - Sink or swim: the Titanic medication administration fair. AB - The Peer Review Committee at a Midwest hospital identified a knowledge deficit relative to medication administration. A continuing increase in the number of medication errors helped the committee to address the issue in a creative and educational way that reinforced knowledge of medication administration. Under the guidance of the Director of Education, employees who recently made medication errors developed and implemented a creative medication administration learning opportunity for hospital employees. The employees chose a project theme, developed educational workstations, and used creative approaches to increase awareness of medication administration pitfalls. This article explains the process of implementing and delivering this fun and exciting learning activity. Theme-based educational experiences are effective teaching strategies that can be used to trigger participants' learning in almost any setting. PMID- 18429373 TI - Becoming and being a preceptor: a phenomenological study. AB - Preceptorship is a valuable component of nursing courses today and is seen as vital to the professional preparation of student nurses. Preceptors facilitate the development of knowledge, clinical skills, and professional attitudes in nursing through guidance, supervision, role modeling, and personal development of the student. They also help to orient and socialize the student to the real nursing workplace environment. Being a preceptor in nursing has been identified as time-consuming and requiring clinical teaching skills that many registered nurses (RNs) perceive they do not possess. This article outlines how a small group of RNs from one selected workplace developed their preceptor abilities through undertaking the preceptor program run by Avondale College to prepare them for this important role. PMID- 18429374 TI - The demographic tidal wave. Are we ready? PMID- 18429375 TI - Over-the-counter medication use in older adults. AB - Use of over-the-counter (OTC) medications is becoming more of a problem in the older adult population as the push to deregulate prescription medications grows. This article summarizes the side effects, adverse reactions, and medication interactions older adults face when using some common OTC medications. PMID- 18429376 TI - Care of community-dwelling older adults by faith community nurses. AB - In this state-funded study of the impact of faith community nursing, 1,061 notes regarding 713 older adults were retrieved from 75 faith community nurses. Critical contacts were mapped using the Data, Interpretation, Action, Response, and Yield (DIARY) charting format. DIARY notes revealed that disabilities were seemingly avoided when reluctant older adults were coaxed to access care before their illnesses escalated. The nurses solicited help from blended networks, procured assistive devices, cued family members, made environments safe, reengaged those who were despairing, calmed those who were anxious, facilitated recovery aftercare, advocated for those who were neglected, and helped caregivers navigate end-of-life decisions. PMID- 18429377 TI - Effects of tangible social support and depression on diabetes self-efficacy. AB - Although social support has been found to promote health and decrease depression in individuals with diabetes, little research has examined the kinds and sources of support. This descriptive correlational study examined the effects of diabetes related tangible social support and depression on diabetes self-efficacy in Hispanic older adults. Participants were predominantly Puerto Rican, and many were depressed. Primary support needs included transportation and communication; family was the primary source of support. Variables associated with diabetes self efficacy included tangible support and education. Understanding the relationship between diabetes tangible support, depression, and diabetes self-efficacy will help nurses adapt their care. PMID- 18429378 TI - A revised Index for Social Engagement for long-term care. AB - The objective of this study was to improve validity and reliability estimates of the Index for Social Engagement (ISE) for long-term care. After exploring content validity and internal consistency in Dutch and Canadian data, two ISE items were dropped, and two new items were added. Reliability of this Revised ISE (RISE) was tested in 189 nursing home residents. It appeared that the RISE has enhanced reliability estimates, especially in residents with cognitive impairment. The RISE for long-term care improves the existing index by including additional dimensions of social engagement and by increasing the reliability of results for residents with cognitive impairment. PMID- 18429379 TI - Nurses' evaluation of the Confusion Assessment Method: a pilot study. AB - Delirium is frequently overlooked in hospitalized adult patients of all ages. Because nurses spend significant amounts of time with patients, the nursing assessment is vital to the identification of delirium. This pilot study investigated nurses'evaluation of the Confusion Assessment Method related to ease of use and helpfulness in identifying delirium in adult patients on two hospital units. Delirium rates are reported for patients across all shifts for 1 month. The majority of nurses found the Confusion Assessment Method to be an effective, user-friendly instrument for the assessment of patients' cognitive status over time. PMID- 18429380 TI - Chemical-specific health consultation for chromated copper arsenate chemical mixture: port of Djibouti. AB - The Agency for Toxic Substances and Disease Registry (ATSDR) prepared this health consultation to provide support for assessing the public health implications of hazardous chemical exposure, primarily through drinking water, related to releases of chromated copper arsenate (CCA) in the port of Djibouti. CCA from a shipment, apparently intended for treating electric poles, is leaking into the soil in the port area. CCA is a pesticide used to protect wood against decay causing organisms. This mixture commonly contains chromium(VI) (hexavalent chromium) as chromic acid, arsenic(V) (pentavalent arsenic) as arsenic pentoxide and copper (II) (divalent copper) as cupric oxide, often in an aqueous solution or concentrate. Experimental studies of the fate of CCA in soil and monitoring studies of wood-preserving sites where CCA was spilled on the soil indicate that the chromium(VI), arsenic and copper components of CCA can leach from soil into groundwater and surface water. In addition, at CCA wood-preserving sites, substantial concentrations of chromium(VI), arsenic and copper remained in the soil and were leachable into water four years after the use of CCA was discontinued, suggesting prolonged persistence in soil, with continued potential for leaching. The degree of leaching depended on soil composition and the extent of soil contamination with CCA. In general, leaching was highest for chromium(VI), intermediate for arsenic and lowest for copper. Thus, the potential for contamination of sources of drinking water exists. Although arsenic that is leached from CCA-contaminated soil into surface water may accumulate in the tissues of fish and shellfish, most of the arsenic in these animals will be in a form (often called fish arsenic) that is less harmful. Copper, which leaches less readily than the other components, can accumulate in tissues of mussels and oysters. Chromium is not likely to accumulate in the tissues of fish and shellfish. Limited studies of air concentrations during cleanup of CCA contaminated soil at wood- preserving sites showed that air levels of chromium(VI), arsenic and copper were below the occupational standards. Workers directly involved in the repackaging, containment or cleanup of leaking containers of CCA or of soil saturated with CCA, however, may be exposed to high levels of CCA through direct dermal contact, inhalation of aerosols or particulates and inadvertent ingestion. Few studies have been conducted on the health effects of CCA. CCA as a concentrated solution is corrosive to the skin eyes and digestive tract. Studies of workers exposed to CCA in wood-preserving plants have not found adverse health effects in these workers, but the studies involved small numbers of workers and therefore are not definitive. People exposed to very high levels of CCA, from sawing wood that still had liquid CCA in it or from living in a home contaminated with ash containing high levels of chromium(VI), arsenic and copper, experienced serious health effects including nosebleeds, digestive system pain and bleeding, itching skin, darkened urine, nervous system effects such as tingling or numbness of the hands and feet and confusion, and rashes or thickening and peeling of the skin. These health effects of the mixture are at least qualitatively reflective of the health effects of the individual components of CCA (arsenic, chromium(VI) and copper). For a given mixture, the critical effects of the individual components are of particular concern, as are any effects in common that may become significant due to additivity or interactions among the components. Effects of concern for CCA, based on the known effects of the individual components, include cancer (arsenic by the oral route, arsenic and chromium(VI) by the inhalation route), irritant or corrosive effects (all three mixture components), the unique dermal effects of arsenic, neurologic effects (arsenic and chromium(VI), and hematologic, hepatic and renal effects (all three components). Because arsenic, chromium(VI), and copper components affect some of the same target organs, they may have additive toxicity toward those organs. Few studies have investigated the potential toxic interactions among the components (arsenic, chromium(VI) and copper) of CCA. The available interaction studies and also possible mechanisms of interaction were evaluated using a weight-of-evidence approach. The conclusion is that there is no strong evidence that interactions among the components of CCA will result in a marked increase in toxicity. This conclusion reflects a lack of well designed interaction studies as well as uncertainties regarding potential mechanisms of interaction. Confidence in the conclusion is low. Workers exposed to high levels of CCA during cleanup of leaking containers of CCA or soil heavily contaminated with CCA should wear protective clothing and respirators if air concentrations of arsenic are above 10 microg/m3. In addition, they should not eat, drink or use tobacco products during exposure to CCA, and should thoroughly wash after skin contact with CCA and before eating, drinking, using tobacco products or using restrooms. When protective clothing becomes contaminated with CCA, it should be changed, and the contaminated clothing should be disposed off in a manner approved for pesticide disposal. Workers should leave all protective clothing, including work shoes and boots, at the workplace, so that CCA will not be carried into their cars and homes, which would endanger other people. People not involved in the cleanup of the CCA and who are not wearing protective clothing should be prevented from entering contaminated areas. Leaking containers of CCA must be repackaged and contained to prevent direct exposure of on-site personnel; and contaminated soil needs to be removed to prevent the CCA from leaching into surface water and groundwater, thereby contaminating sources of drinking water. PMID- 18429381 TI - Ebselen protects brain, skin, lung and blood cells from mechlorethamine toxicity. AB - Nitrogen mustards are vesicants capable of burning the skin, eyes and respiratory tract of exposed individuals. While generally less toxic than sulfur mustards, these compounds have the potential for use as chemical warfare agents. Presently, no antidote exists for treatment against nitrogen mustard toxicity. The purpose of this study was to investigate the in vitro toxicity of the nitrogen mustard mechlorethamine (HN2) in four cell models: CEM-SS human T cells, A431 human skin epithelial cells, rat hippocampal astrocytes and rat pleural mesothelial cells. Furthermore, the efficacy of the synthetic seleno-organic compound ebselen (Eb) (2-phenyl-1,2- benzisoselenazol-3(2H)-one) as a cytoprotective agent against such toxicity was evaluated. Significant increases in cell viability, as assessed using an MTT assay for viability, was demonstrated when 30 microM Eb was used as a cotreatment with HN2 in all cell models tested at the following doses of HN2: A431 skin cells,10-40 microM; rat astrocytes, 20 and 40 microM; rat mesothelia, 10-40 microM; and human T cells 4-16 microM. Decreases in cell viability and toxicity to HN2 were confirmed using light and scanning electron microscopy. Membrane damage, observed with HN2 exposure, such as blebbing and loss of cell projections, was ameliorated with Eb cotreatment. Our results demonstrate a generalized protective effect observed with Eb cotreatment that suggests that this agent may have potential as an antidote for HN2 exposure and toxicity. PMID- 18429382 TI - JP-8 jet fuel exposure rapidly induces high levels of IL-10 and PGE2 secretion and is correlated with loss of immune function. AB - The US Air Force has implemented the widespread use of JP-8 jet fuel in its operations, although a thorough understanding of its potential effects upon exposed personnel is unclear. Previous work has demonstrated that JP-8 exposure is immunosuppressive. In the present study, the potential mechanisms for the effects of JP-8 exposure on the immune system were investigated. Exposure of mice to JP-8 for 1 h/day resulted in immediate secretion of two immunosuppressive agents; namely, interleukin-10 (IL-10) and prostaglandin E2 (PGE2). JP-8 exposure rapidly induced a persistently high level of serum IL-10 and PGE2 at an exposure concentration of 1000 mg/m3. IL-10 levels peaked at 2 h post-JP-8 exposure and then stabilized at significantly elevated serum levels, while PGE2 levels peaked after 2-3 days of exposure and then stabilized. Elevated IL-10 and PGE2 levels may at least partially explain the effects of JP-8 exposure on immune function. Elevated IL-10 and PGE2 levels, however, cannot explain all of the effects due to JP-8 exposure (e.g., decreased organ weights and decreased viable immune cells), as treatment with a PGE2 inhibitor did not completely reverse the immunosuppressive effects of jet fuel exposure. Thus, low concentration JP-8 jet fuel exposures have significant effects on the immune system, which can be partially explained by the secretion of immunosuppressive modulators, which are cumulative over time. PMID- 18429383 TI - Abdominal bloating and irritable bowel syndrome like symptoms following microinstillation inhalation exposure to chemical warfare nerve agent VX in guinea pigs. AB - While assessing the methylphosphonothioic acid S-(2-(bis(1 methylethyl)amino)ethyl)O-ethyl ester (VX) induced respiratory toxicity and evaluating therapeutics against lung injury, we observed that the animals were experiencing abnormal swelling in the abdominal area. Nerve agent has been known to increase salivary, nasal and gastrointestinal secretion and cause diarrhea. This study was initiated to investigate the effect of VX on the gastrointestinal tract (GI) since abdominal pathology may affect breathing and contribute to the on going respiratory toxicity. The mid-abdominal diameter and the size of the lower left abdomen was measured before and after 27.3 mg/m3 VX exposure by microinstillation and at 30 min intervals up to 2 h post-VX exposure. Both VX and saline exposed animals exhibited a decrease in circumference of the upper abdomen, although the decrease was slightly higher in VX-exposed animals up to 1 h. The waist diameter increased slightly in VX-exposed animals from 60 to 90 min post-VX exposure but was similar to saline controls. The lower left abdomen near to the cecum, 6 cm below and 2cm to the right of the end of the sternum, showed an increase in size at 30-60 min that was significantly increased at 90-120 min post-VX exposure. In addition, VX-exposed animals showed loose fecal matter compared to controls. Necropsy at 24h showed an increased small intestine twisting motility in VX-exposed animals. Body tissue AChE assay showed high inhibition in the esophagus and intestine in VX-exposed animals indicating that a significant amount of the agent is localized to the GI following microinstillation exposure. These results suggest that microinstillatipn inhalation VX exposure induces gastrointestinal disturbances similar to that of irritable bowel syndrome and bloating. PMID- 18429384 TI - [The comprehensive evaluation of the public health system effectiveness: the methodological approach and the criteria]. AB - The issue deals with the comprehensive evaluation of the public health system effectiveness. The criteria assessing the medical economic and medical social effectiveness are discussed. The resultfulness of health care management is considered. The estimation techniques, the mathematical modeling and the economic analysis of health capital losses are proposed. PMID- 18429386 TI - [The results of the study of the medical social characteristics of women in childbirth]. PMID- 18429385 TI - [The risk of infectious morbidity among population under the pollution of the sanitary protective and recreation areas of water consumption in the city of Krasnodar]. AB - The probability of development of public health derangements among population due to the anthropogenic pollution of the environment in the most cases depends on specific public, social economic and other characteristics of the community on the investigated territory. In the article the data related to the infectious morbidity developed because of water factor and due to the pollution of the soil and water in the sanitary protective and recreation areas of water consumption in the city of Krasnodar is presented. The mentioned recommendations include the organizational and medical preventive measures targeted to the optimization of the industrial and domestic water consumption and to the elimination the causes of the pollution of drinking water with xenobiotics in the centralized water supply system. PMID- 18429387 TI - [The characteristics of the reproductive behavior of women depending on the social hygienic particularities of their families]. PMID- 18429388 TI - [The system of forecasting the health disorders among the adolescents at the age of 15-17 years]. AB - The issues of forecasting of health disorders among adolescents at the age of 15 17 years are discussed. The analysis and assessment of the factors of biological and social anamnesis of the adolescents is carried out. The psychological and functional risk factors are revealed. The tables to make up the forecasting of the development of health disorders among adolescents are developed. It is proved that the forecasting of health disorders among adolescents is to be implemented considering the successfulness of their education. PMID- 18429389 TI - [On the procedures of preventing the health losses of adolescents under the conditions of rural area]. AB - To implement the effective management of the system of prevention of the health losses among adolescents under the conditions of rural area the levels of preventive activities were established. The criteria of assessment of procedures applied to prevent the health losses among the adolescents under the conditions of rural area were developed. The new approaches to the integrated intersectoral indicators as applied to the prevention of pathology among adolescents brought to the light its inadequate quality level and approved the need of the consolidation of the intersectoral structures in this field. PMID- 18429390 TI - [The population morbidity and the issues of medical prevention of the municipal level]. PMID- 18429391 TI - [The dynamics of the indicators of morbidity of the diseases of musculoskeletal system and connective tissue as related to the mode of the population orthopedic care]. PMID- 18429392 TI - [The medical social characteristics of reproductive health and reproductive behavior of military personnel on legal contract]. AB - The medical social characteristics of reproductive health of armed forces personnel on legal contract are established. Among them are higher prevalence of contact with professional health hazards, unsatisfactory welfare standards and poor housing conditions and higher level of propagation of pernicious habits. The reproductive behavior of armed forces personnel is characterized as destructive one. It is established that part of males have negative reproductive attitude and that the attitude to the family with fewer children is prevailing. PMID- 18429393 TI - [The medical psychological monitoring of the conscript contingent]. PMID- 18429394 TI - [The factors affecting the quality of life of patients with stroke aftermaths]. AB - The characteristics of patient caring after the stroke enhancing the effectiveness of the rehabilitation treatment and the social and everyday adaptation as well are discussed. Key words include the stroke, the rehabilitation, the quality of life, the physical therapy, the ergo therapy, the psychotherapy, the neuropsychology, the social rehabilitation. PMID- 18429395 TI - [The transfusion service in the multi-type medical institution]. AB - The importance of the transfusion maintenance of the curative process becomes so crucial that it needs the institution of specialized service. The practical experience and the analysis of the material testify the necessity of the separate transfusion service in the multi-type medical institution. Its structural and functional characteristics are discussed. Such kind of approach allows to optimize the provision of the patients with the high-quality and virus-free blood components. These measures result in the positive economic effect due to the rational use of resources, decreasing of time period of patients stay in the hospital and enhancement of therapy effectiveness. PMID- 18429396 TI - [The actual issues of medical service pricing in allergodermatology]. AB - The article is devoted to the issues of the medical service price formation in the allergodermatology. The competent and proper pricing of provided medical services promotes the development of the institution, the enhancement of its effectiveness and the quality of its services. The information of research study organized in the provincial health institution of the Sverdlovsk oblast is presented. PMID- 18429397 TI - [The maintenance of security of the elderly and gerontic patients in the hospital conditions]. PMID- 18429398 TI - [The state and the perspectives of the secondary medical education in the Orenburg oblast]. AB - The article is dedicated to the issues of the actual state and the perspectives of the secondary medical education in the Orenburg oblast in the framework of the establishment of the primary medical sanitary care. It is emphasized that to enhance the secondary medical education under the new economical conditions it is needed to revise the actual educational programs of training and re-training of paramedical personnel and to adjust them to the up-to-date level. The implementation of the new approaches to the standards in the system of primary medical sanitary care and to the modernization of the educational programs for the paramedical personnel. PMID- 18429399 TI - [The historical aspects of rural public health]. PMID- 18429400 TI - [V. I. Gedreitz as the professor of surgery and the writer]. PMID- 18429401 TI - [On the history of the activities of the Red Cross Court Hospital for lower rank officials (1915-1917)]. PMID- 18429402 TI - [The public administration of medicine in the ancient China]. PMID- 18429403 TI - [Epidemiology of bluetongue virus serotype 8 outbreaks in the Netherlands in 2006]. AB - In August 2006 a major epidemic of Bluetongue (BT) occurred in north-western Europe, affecting The Netherlands, Belgium, Germany, Luxemburg, and the north of France. It was caused by Br virus serotype 8 (BTV-8), a serotype previously unknown to the EU. Although clinical disease is usually restricted to sheep, this virus also caused clinical disease in a small proportion of cattle. The last clinical outbreak of BT in The Netherlands occurred mid-December 2006. The delay between observation of the first clinical signs by the owner and reporting of a clinically suspect BT situation to the veterinary authorities was approximately 2 weeks. BTV-8-associated clinical signs were more prominent in sheep than in cattle, and the relative frequency of specific clinical signs was different in cattle and sheep. Morbidity and mortality rates were significantly higher among sheep than among cattle, and a higher proportion of cattle than sheep recovered from clinical disease. PMID- 18429404 TI - [Phenylbutazone, the black 'horse' of the NSAID family?]. PMID- 18429405 TI - [Sunday's veterinarian]. PMID- 18429406 TI - [KNMVD position on unanesthetized slaughtering of animals]. PMID- 18429407 TI - [Transportation of animals and emergency slaughtering: what are the rules?]. PMID- 18429408 TI - [Coaching in veterinary practice]. PMID- 18429409 TI - Addressing resistance to recovery. PMID- 18429410 TI - 'A true role model'. PMID- 18429411 TI - Changing times but a constant mission. PMID- 18429412 TI - Making your messages 'stickier'. PMID- 18429413 TI - It's the bounce that counts. PMID- 18429415 TI - Giving clients MORE after treatment. PMID- 18429414 TI - Hope for reentering the community. PMID- 18429417 TI - Beyond the blueprints. PMID- 18429416 TI - No library card needed. PMID- 18429418 TI - Creating environments for modern care. PMID- 18429419 TI - Understanding standards development. PMID- 18429420 TI - Some people need help to get help. PMID- 18429421 TI - Making the case for mental healthcare. PMID- 18429422 TI - The use of implants and ovate pontics in the esthetic zone. AB - This article will discuss and evaluate the potential conditions that can present in patients who require or already have had multiple anterior tooth extractions; the proper considerations for the use of ovate pontics in the treatment plan also will be discussed. While the ultimate treatment decisions must be determined on a case-by-case basis, it is important to recognize in advance the various potential outcomes to ensure that realistic decisions are made about the best treatment options for each patient. Accordingly, the four most common presentations a clinician is likely to encounter will be examined, as well as how they may be managed and the most likely compromises that might exist in the final result. PMID- 18429423 TI - Fluoride-releasing materials and their adhesive characteristics. AB - This article discusses fluoride-releasing materials, including their caries inhibiting properties and clinical longevity, as well as provides useful application tips. Based on results from clinical trials, there is growing evidence that fluoride-releasing materials, particularly glass ionomers, reduce the occurrence of recurrent caries. As such, they should be employed-especially in the case of high caries-risk patients-as part of an overall treatment plan for maximum care. PMID- 18429424 TI - Bisphosphonate-related osteonecrosis of the jaws. AB - Bisphosphonates are a class of agents whose efficacy in treating and preventing the skeletal complications associated with osteoporosis and malignant bone metastases has been well established. Despite this benefit, osteonecrosis of the jaws is a significant complication in a subset of patients receiving these drugs. Based on a growing number of case reports and institutional reviews, bisphosphonate therapy may cause bone to become exposed and necrotic. Currently, this phenomenon is isolated to the jaw. This complication usually presents following simple dentoalveolar surgery. The pathogenesis for this complication appears to be related to the profound inhibition of osteoclast function and bone remodeling. This article serves to alert ists and dental specialists about the potential complication of jaw necrosis in patients receiving bisphosphonate therapy, and proposes a guideline for diagnosis, staging and management. PMID- 18429425 TI - Ridge-splitting technique with simultaneous implant placement. AB - Bone augmentation procedures are routinely required before dental implant placement. Several techniques for this procedure may be considered, such as guided bone regeneration, bone block grafting, and ridge splitting for bone expansion. These case reports describe the technique for ridge splitting and gradual expansion in the maxilla and the mandible. Simultaneously, dental implants were placed within the split ridge, surrounded by the particulate bone graft and covered by a resorbable membrane. Six months later, the implants were uncovered followed by impression and final restoration with implant-supported porcelain-fused-to-metal crowns. PMID- 18429426 TI - Bisphosphonate-induced osteonecrosis: dental considerations. PMID- 18429428 TI - It's a privilege. The board's role in physician credentialing and privileging. AB - What are the board's responsibilities for physician credentialing and privileging? With new standards from the Joint Commission and a growing emphasis on improving executive/physician relationships, the board needs a deeper understanding of its role. PMID- 18429427 TI - The healing socket and socket regeneration. AB - Tooth extraction is a common procedure in dentistry. The normal healing response to the procedure results in a significant loss of bone and collapse of the surrounding gingiva. In addition to normal healing, a substantial percentage of extraction sites suffer postoperative complications. This article presents histology that supports the concept that the first response to extraction is bone death and resorption of the socket wall. The stages of extraction socket healing also will be discussed. Additionally, the article will present a regenerative method that skips the resorptive phase, the clotting phase, the granulation of tissue phase, and the collagen-producing phase of normal extraction-socket healing, while avoiding extraction-socket complications. PMID- 18429429 TI - Never land. Medicare declares 'no pay for preventable errors'. AB - Preventing medical errors has become a front-burner issue for hospitals over the last decade, but Medicare has just turned up the heat with its new policy of not paying for preventable injuries and errors. PMID- 18429430 TI - The keys to nurse staffing. PMID- 18429431 TI - Hospitals weigh in on quality. AB - The board's role in quality oversight is growing more important every day, not only because it's the right thing to do, but also because pay for performance and increasing demands for transparency are requiring more accountability. PMID- 18429432 TI - Rural hospitals: a different take on conflict of interest. PMID- 18429433 TI - In troubled waters, hospitals need an able captain. PMID- 18429435 TI - Leaders listen, respond to nurses' woes. PMID- 18429434 TI - Monitoring patient safety and quality: a simple framework. PMID- 18429436 TI - Preferential suppression of yellow head virus (YHV) envelope protein gp116 in shrimp that survive challenge with YHV. AB - The DNA sequence that encodes the first 406 amino acid residues at the N-terminus of yellow head virus (YHV) protein gp116, namely N/2 gp116deltaTM, and the DNA sequence that encodes the next 392 amino acid residues at the C-terminus of gp116 (without the transmembrane region), namely C/2 gp116deltaTM, were cloned into pGEX-6P-1 plasmid and expressed in E. coli. Both recombinant proteins were expressed, purified by SDS-PAGE and used to immunize mice. The mouse anti recombinant N/2 gp116 and C/2 gp116 antisera bound specifically to both the recombinant proteins and to natural gp116 protein in YHV-infected haemolymph as shown by Western blotting and in tissues as shown by immunohistochemistry. Immunohistochemical localization of YHV using anti-gp116 antiserum or monoclonal antibodies specific to gp116 (V3-2B), gp64 (Y18) and p20 (Y19) revealed similar immunoreactivity patterns for all these reagents in muscle and mandibular tissue in shrimp showing gross signs of yellow head disease. However, in gill, hepatopancreas, lymphoid organ and thoracic ganglion tissues from experimental YHV-infected shrimp (Penaeus vannamei and Palaemon serrifer) that did not show signs of disease, immunoreactivity to gp116 was reduced or absent while that for gp64 and p20 remained intense. Thus, some shrimp species were able to selectively inhibit the synthesis of gp116 in a manner that was associated with absence of gross signs of disease. PMID- 18429437 TI - Lactococcus lactis subspecies lactis also causes white muscle disease in farmed giant freshwater prawns Macrobrachium rosenbergii. AB - From May to August 2001 in Taiwan, 27 farms for the giant freshwater prawn Macrobrachium rosenbergii experienced white tail disease outbreaks in animals approximately 3 to 5 mo old, with total lengths from 6 to 8 cm. Examination of the infected prawns revealed not only previously reported Lactococcus garvieae (16 farms) but also the novel L. lactis subsp. lactis (10 farms). One farm had shrimp infected with both bacteria. In the farms with L. lactis infections, the cumulative mortality was approximately 25 to 60%. Gross signs of disease were opaque and whitish muscles, while histopathology included marked edema and necrotic lesions, with inflammation in the muscles and hepatopancreas. Bacteria isolated using brain/heart infusion medium or tryptic soy agar were Gram-positive and ovoid. Eleven isolates from different farms were identified as L. lactis subsp. lactis using API 20 Strep and Rapid ID32 Strep tests and using PCR assays specific for the L. lactis subsp. lactis 16S rDNA gene (650 bp amplicon) and for the 16S to 23S rDNA interspacer region (380 bp amplicon). In addition, sequencing of the full 16S rDNA genes of 2 of the isolates (MR17 and MR26; GenBank Accession Numbers AF493058 and AF493057, respectively) revealed 99.9% identity between the isolates and 98.7% identity to several complete 16S rRNA sequences of L. lactis subsp. lactis at GenBank. Experimental infections with our isolates gave gross signs and histopathological changes similar to those seen in naturally infected prawns. The mean lethal dose of 4 isolates and the reference strain L. lactis subsp. lactis BCRC 10791 ranged from 4.2 x 10(6) to 2.5 x 10(7) colony-forming units prawn(-1), indicating virulence similar to that previously reported for L. garvieae. This is the first report confirming L. lactis subsp. lactis as a pathogen in juvenile and adult prawns from aquaculture. PMID- 18429438 TI - Enhanced propagation of fish nodaviruses in BF-2 cells persistently infected with snakehead retrovirus (SnRV). AB - Fish nodaviruses are causative agents of viral nervous necrosis causing high mortality in cultured marine fishes around the world. The first successful isolation of fish nodavirus was made with SSN-1 cells, which are persistently infected with snakehead retrovirus (SnRV). In the present study, a BF-2 cell line persistently infected with SnRV (PI-BF-2) was established to evaluate the influence of SnRV on the production of fish nodavirus. The PI-BF-2 cells were slightly more slender than BF-2 cells, but no difference was observed in propagation rate between both cell lines. No difference was observed in production of SnRV between PI-BF-2 and SSN-1 cell lines. Although both PI-BF-2 and BF-2 cell lines showed no cytopathic effect (CPE) after inoculation of striped jack nervous necrosis virus (SJNNV) and redspotted grouper nervous necrosis virus (RGNNV), these fish nodaviruses could be amplified in BF-2 cells, and moreover, production of fish nodaviruses in the PI-BF-2 cell line was more than 40 times higher than in BF-2 cells. Thus, it was concluded that BF-2 cell permissiveness to fish nodaviruses was enhanced by persistent infection with SnRV. Furthermore, homologous cDNA to genomic RNA of SJNNV was detected from both PI-BF-2 and SSN-1 cell lines persistently infected with SnRV. The amount of nodavirus cDNA in SJNNV-inoculated PI-BF-2 cells was clearly lower than that in SJNNV-inoculated SSN-1 cells. PMID- 18429439 TI - Antigenic and molecular characterization of Vibrio ordalii strains isolated from Atlantic salmon Salmo salar in Chile. AB - Biochemical, serological and molecular properties of a group of 14 Vibrio ordalii strains isolated from cultured Atlantic salmon Salmo salar in Chile in recent years were studied. The characteristics of isolates were compared with the type strain V. ordalii ATCC 33509T. The Chilean V. ordalii represented a biochemically homogenous group; however, some minor differences with the type strain were observed. The serological relationships among isolates, as well as the study of their antigenic determinant (LPS) revealed a strong reaction with antisera raised against Atlantic salmon strains and the antiserum raised against Listonella anguillarum serotype O2. However, LPS electrophoretic patterns were completely different from the V. ordalii type strain, regardless of the serum employed, suggesting the possibility that the Chilean strains constitute a new serological subgroup within this bacterial species. Genetic analyses by PFGE, RAPD, REP-PCR and ERIC-PCR demonstrated that all V. ordalii strains were genetically homogenous, displaying similar DNA patterns, regardless of the techniques used. Moreover, the analysis of DNA banding patterns generated by ERIC-PCR and REP-PCR also clearly separated the type strain from the Chilean strains. This is the first report of characterization of V. ordalii strains from the Southeastern Pacific area, the results of which should facilitate the development of vaccines for protecting cultured Atlantic salmon against vibriosis in this area. PMID- 18429440 TI - Anguillicola crassus infection in Anguilla rostrata from small tributaries of the Hudson River watershed, New York, USA. AB - We studied the invasion of the exotic nematode parasite Anguillicola crassus in the American eel Anguilla rostrata using tributaries of the Hudson River estuary. Yellow-phase American eels were sampled from 6 tributaries, and their swim bladders were examined for nematode infection. Prevalence averaged 39% with an intensity of 2.4 nematodes per eel. Parasite distribution was not significant along a latitudinal gradient; on the other hand, physical barriers (dams and natural waterfalls) significantly reduced infections upstream. Urbanization may increase the susceptibility of eels to infection; we found significantly elevated infection rates when urbanized lands exceeded 15% of the tributary catchment area. Yellow-phase eel condition was not affected by parasite infection. The invasion of the entire Hudson River watershed is ongoing and therefore will continue to be a management concern. Further analysis of the parasite-host interaction in North America is warranted. PMID- 18429441 TI - Demonstrating freedom from Gyrodactylus salaris (Monogenea: Gyrodactylidae) in farmed rainbow trout Oncorhynchus mykiss. AB - This paper describes an approach to demonstrate freedom of individual rainbow trout farms from Gyrodactylus salaris Malmberg, 1957. The infection status of individual farms is relevant should G. salaris be introduced into a country or zone previously known to be free of the parasite. Trade from farms where G. salaris may have been introduced would be restricted until freedom had been demonstrated. Cage, fish and parasite sample sizes were calculated based on the minimum detectable prevalence (P*), test characteristics, population size, and Type I and II errors. Between 5 and 23 cages per farm would need to be sampled to demonstrate freedom at a cage level P* of 10%. The number of fish sampled per cage depended mainly on the test sensitivity (probability of correctly identifying an infected fish). Assuming a test sensitivity of 99% at the fish level, 59 fish per cage are needed (P* = 5%). Since G. salaris may exist in mixed infection with G. derjavini, testing a sample of gyrodactylid parasites may not result in the parasite being detected when present. Test sensitivity at the fish level depends on the number of gyrodactylids on the fish, the proportion of which are G. salaris and the number examined. Assuming a P* of 5% (i.e. G. salaris are at least 5% of the gyrodactylid population), between 20 and 73 parasites per fish would need to be sampled (depending on abundance) to maintain the Type I error at 0.01 (thus a fish level test sensitivity of 99%). This work identifies the critical information, and further research, needed to assess freedom from G. salaris with a known level of confidence; this is essential to provide a sound scientific basis for decision-making about disease control measures. PMID- 18429442 TI - An epizootic of Caligus chiastos on farmed southern bluefin tuna Thunnus maccoyii off South Australia. AB - In some years, large numbers of Caligus chiastos have been observed on the external surfaces of southern bluefin tuna, particularly on the head and eyes, in some sea cages in Spencer Gulf, Australia. As no epidemiological data were available, we monitored sea lice on tuna (N = 130) in 4 research cages sampled at 6 wk intervals during the 2005 farming season. No lice were observed on a sample of 10 wild-caught tuna when the cohort was transferred to cages in early April. By late May more than half the sampled tuna (22 of 40) were infected, with up to 42 parasites; we also recorded one unidentified Caligus sp. at this time. In early July the number of tuna infected with lice declined to 10%; in the final sample in late August none were detected. Prevalence in May was significantly higher than on other dates (p < or = 0.001), whereas mean abundances did not differ significantly (p > 0.05). The decline in prevalence corresponded with a seasonal fall in temperature, from ca. 17 degrees C in May to 14 degrees C in August. Counts of lice at the peak of infection were associated with the severity of eye damage (Spearman's rank correlation coefficient, r(S,38df) = 0.654, p < 0.001); this may be because lice graze on the cornea or because tuna injure their eyes when flashing (rubbing against objects). Counts at this time were also strongly and inversely correlated with the condition index (r(S,38df) = -0.707, p < 0.001). It appears that tuna become infested with adult sea lice via wild teleosts and elasmobranchs attracted to sea cages. PMID- 18429443 TI - Delta de l'Ebre is a natural bay model for Marteilia spp. (Paramyxea) dynamics and life-cycle studies. AB - Marteilia spp. are paramyxean parasites that affect several bivalve species of economic interest, such as Ostrea edulis and Mytilus galloprovincialis. Certain aspects of Marteilia spp., such as their life cycle and host affinity and infection dynamics, still remain unknown. The 'Delta de l'Ebre' constitutes a natural model for the study of the life cycle of the parasite Marteilia, since uninfected mussels and flat oysters immersed in the bays can become infected. This, along with the geographical and ecological characteristics of the bays, make it a very interesting location to study the Marteilia life cycle. Preliminary results concerning marteiliosis, mainly in mussels, such as prevalence dynamics, infectious periods, host affinity and host intermediate candidates are reported in the present paper. This information will be required for further, more exhaustive, studies in the bays of the Ebre delta. PMID- 18429444 TI - Specific monoclonal antibodies raised against Taura syndrome virus (TSV) capsid protein VP3 detect TSV in single and dual infections with white spot syndrome virus (WSSV). AB - The gene sequence encoding VP3 capsid protein of Taura syndrome virus (TSV) was cloned into pGEX-6P-1 expression vector and transformed into Escherichia coli BL21. After induction, recombinant GST-VP3 (rVP3) fusion protein was obtained and further purified by electro-elution before use in immunizing Swiss mice for production of monoclonal antibodies (MAb). One MAb specific to glutathione-S transferase (GST) and 6 MAb specific to VP3 were selected using dot blotting and Western blotting. MAb specific to VP3 could be used to detect natural TSV infections in farmed whiteleg shrimp Penaeus vannamei by dot blotting and Western blotting, without cross reaction to shrimp tissues or other shrimp viruses, such as white spot syndrome virus (WSSV), yellow head virus (YHV), monodon baculovirus (MBV) and hepatopancreatic parvovirus (HPV). These MAb were also used together with those specific for WSSV to successfully detect TSV and WSSV in dual infections in farmed P. vannamei. PMID- 18429446 TI - Seven procedures for a successful, long-lasting reprocessing program. PMID- 18429445 TI - Virus-like particles in Urastoma cyprinae, a turbellarian parasite of Mytilus galloprovincialis. AB - Urastoma cyprinae is a turbellarian parasite infecting the gills of several marine bivalves. We observed the presence of virus-like particles (24 to 30 nm in diameter; icosahedral symmetry) arranged in paracrystalline arrays in the cytoplasm of subepidermal dorsal cells. These particles appear to be RNA viruses, possibly related to the Picornaviridae. This is the first report of viral particles in the parasite U. cyprinae. PMID- 18429447 TI - Development of anti-infective, specialized devices likely to influence future of Foley catheters. PMID- 18429448 TI - Passing the torch: prepare today for tomorrow. PMID- 18429449 TI - We have come so far. PMID- 18429450 TI - Ten years on, have Blair's 'super nurses' delivered for patients? Interview by Sarah Harrison. PMID- 18429451 TI - Which areas are in the greatest need of mental health services? PMID- 18429452 TI - A healthier harvest. AB - Fresh air and fresh food are all part of a Rethink allotment project to help improve the health and wellbeing of people with mental illness. PMID- 18429453 TI - Listen and learn. AB - A buddy scheme is helping nursing and midwifery students to gain valuable insight into mental illness. PMID- 18429454 TI - Passion for prescribing. PMID- 18429455 TI - Know your patient. PMID- 18429456 TI - Writing for publication: a practical guide. AB - Nurses in clinical practice tend not to view writing for publication as part of their role, and often fail to recognise the value of written communication as a means of sharing valuable knowledge and expertise with others. The tendency to view writing for publication as an arduous and daunting task, reserved only for those in academia, may deter the novice writer. This article aims to encourage nurses in clinical practice to consider writing for publication and actively contribute to professional development through the dissemination of nursing knowledge. The article also provides a practical guide for writing a research and quality improvement article. PMID- 18429457 TI - Care of patients who have died. AB - This article outlines the steps involved in preparing the patient after death- known as last offices. It addresses legal and non-legal issues, religious considerations, hygiene care and aftercare for the family. PMID- 18429458 TI - Understanding schizophrenia. AB - This article provides an introduction to schizophrenia, outlining its signs and symptoms, causes and possible prognosis for patients. It also considers a range of therapeutic approaches to treatment. PMID- 18429459 TI - Prostate cancer. PMID- 18429460 TI - Make a difference. PMID- 18429461 TI - Power of the team. PMID- 18429462 TI - Lean and simple. PMID- 18429463 TI - Bracing for the storm. PMID- 18429464 TI - The case against CON laws. PMID- 18429465 TI - Physicians. Rise in specialists puts general surgery in a bind. PMID- 18429466 TI - Emergency department. Pharmacists in the ED help reduce errors. PMID- 18429467 TI - Regulation. States, hospitals worry that CMS directive will hinder coverage. PMID- 18429468 TI - Other voices. New leapfrog leader says health care lacks common sense. PMID- 18429469 TI - Practical ethics. Can you grow and still preserve? PMID- 18429470 TI - Storyboard. Steps to better board accountability. PMID- 18429471 TI - Finance. Health care may not be immune from the economic downturn. PMID- 18429472 TI - Challenges, not excuses. PMID- 18429473 TI - Your future chief of staff? AB - Jay Parkinson, M.D., is among the new breed of physicians focusing on Generation Y, young consumers whose assumptions about life could dramatically change health care. They're technically savvy, highly social, very independent--and they refuse to accept the status quo. Parkinson has built an officeless practice based on e mails, cell phones and the Web. Some observers say other providers, including hospitals, should take heed. PMID- 18429474 TI - OR of the future: it's here and now. AB - It sounds grandiose, but those involved in the Operating Room of the Future say hospitals with limited resources can adapt its methods to improve productivity. PMID- 18429475 TI - Hospital construction roars right along. AB - The costs of building and renovation keep climbing, but nobody expects the hospital construction boom to end soon. In fact, it's doubled in two years. PMID- 18429476 TI - How hospitals can prepare for the new MS-DRGs. AB - The Centers for Medicare & Medicaid Services is rolling out the most significant payment changes to Medicare since 1983. Here's what it means for hospitals. PMID- 18429477 TI - The 2007 Foster McGaw Prize. Harborview Medical Center's community services address an astounding variety of needs. AB - Harborview Medical Center's community programs address an astounding variety of needs in Seattle. It won this year's $100,000 prize. PMID- 18429478 TI - Data page: Survey: trustees can expect more scrutiny. PMID- 18429479 TI - Health care: getting what you pay for. PMID- 18429480 TI - HIV testing in the ED. PMID- 18429481 TI - Physical activity and psychological well-being among Hong Kong Chinese older adults: exploring the moderating role of self-construal. AB - This study examined the association between physical activity (PA) and psychological well-being--self-esteem and relatedness satisfaction--among 102 Hong Kong Chinese older adults. It also tested whether independent-interdependent self-construal moderated the association. Physical activity, self esteem, relatedness satisfaction, and self-construal were measured by the Yale Physical Activity Scale, the Rosenberg's Self-Esteem Scale, the Relatedness Satisfaction subscale of the Basic Need Satisfaction in General Questionnaire, and Gudykunst, Matsunoto, Ting-Toomey, and Nishida's (1996) Self-Construal Scale respectively. Findings revealed a positive association between PA and relatedness satisfaction, but not self-esteem. Interdependent self-construal moderated the relationship between physical exercise and relatedness satisfaction. These findings suggest that PA may be psychologically beneficial to older adults across cultures, but its benefits may depend on cultural emphasis and individual self-construal. PMID- 18429482 TI - The relationship between grandparent satisfaction, meaning, and generativity. AB - This study investigated predictors of grandparent satisfaction. Participants were 149 non-custodial grandparents (100 grandmothers, 49 grandfathers) aged up to 80 years. They completed grandparent satisfaction, meaning, and generativity scales, and indicated levels of weekly childcare contact with grandchildren. As expected, increasing levels of generativity predicted greater satisfaction, as did valued elder and centrality meanings. Unexpectedly, reinvolvement, immortality, and indulgence meanings were not predictors, nor was childcare contact. There was only partial support for grandparent gender as a predictor: being a grandmother predicted greater satisfaction, but this effect disappeared when the meaning and generativity variables were included in the analysis. In conclusion, the findings highlight the significance the role can have for many grandparents. Grandparenting can be satisfying because it provides a sense of purpose, and feeling of being valued during middle and later life when generative developmental needs are greatest. PMID- 18429483 TI - Normative beliefs about sharing housing with an older family member. AB - The aims of this study were: (a) to examine general perceptions of filial obligations toward sharing housing with older parents and stepparents; and (b) to assess the effects of selected contextual factors on those normative beliefs. A national sample of 579 men and 582 women (mean age = 44.6, SD = 17.2) responded to a multiple segment factorial vignette in which an older parent or stepparent was portrayed as needing help with housing. Respondents thought that parents should be helped more than stepparents, younger adults with greater resources were more obligated to help older parents and stepparents than were those with meager resources, and older parents and stepparents with greater need acuity were expected to be helped more than older parents and stepparents with less serious housing needs. Attitudes about co-residence were based on family obligation norms, beliefs about repaying older adults for past help, perceived relationship quality, other demands on the younger adult's resources, the older person's resources, and moral responsibilities to assist. PMID- 18429484 TI - The link between successful aging and serious leisure. AB - The purpose of this qualitative study is to examine whether engagement in a serious leisure activity provided older adults opportunities for successful aging. Data were collected through in-depth interviews at shag dance festivals in North Myrtle Beach, South Carolina. To provide structure for the interview, a general interview guide consisting of open-ended questions was used. A total of 31 interviews were conducted with 25 individuals and 6 couples, between 60 and 82 years of age. During the analysis stage, the constant comparative method was used to analyze the data. Themes that emerged support the idea that shag dancing is a form of serious leisure for older adults and support the role of serious leisure in successful aging. The narratives provided by the informants illustrate the powerful role serious leisure can play in an individual's life and point toward the value of serious leisure as an approach that holds promise for recreators working to help older individuals experience successful aging. PMID- 18429485 TI - Meet me at the (West Coast) Hotel: the Lochner era and the demise of Roe v. Wade. PMID- 18429486 TI - [Genetic variation and recombination have profound medical implications]. AB - In the family Herpesviridae, the subfamily Alphaherpesvirinae contains numerous pathogenic viruses, i.e. herpes simplex and varicella-zoster viruses. These double-stranded DNA viruses exhibit a complex cycle combining lytic and latent infections. Moreover, both intranuclear replication and a sophisticated DNA replication machinery allow an efficient proof-reading mechanism of correction. A low mutation rate is therefore encountered by these viruses. Recombination can be identified as a key element of the genetic biodiversity of alphaherpesviruses, together with mutations. The experimental data recently obtained in the bovine herpesvirus 1 homologous model support the importance of recombination in alphaherpesvirus evolution and its role in the mechanisms involved by the virus to escape from medical tools of prevention and treatment. PMID- 18429487 TI - [Human resistance to African trypanosoma infections]. AB - African trypanosomes (prototype: Trypanosoma brucei) are protozoan flagellates that infect a wide range of different mammals. In humans, these parasites have to counteract innate immunity because human serum possesses efficient trypanolytic activity. Resistance to this activity has arisen in two T. brucei subspecies, termed T. b. rhodesiense and T. b. gambiense, allowing them to infect humans where they cause sleeping sickness in East and West Africa respectively. The study of the mechanism by which T. b. rhodesiense escapes lysis by human serum led to the identification of the trypanolytic factor, which turned out to be an ionic pore-forming apolipoprotein associated with some HDL particles. PMID- 18429488 TI - [Congenital Chagas disease: from the laboratory to public health]. AB - Trypanosoma cruzi, the protozoan agent of Chagas disease can be transmitted from mother to foetus. The incidence of congenital infection is estimated to be at least 15,000 cases per year in Latin-America. Its incidence in the non endemic countries (U.S.A., Europe, Japan) is not known. Thanks to multidisciplinary studies performed in Bolivia, it has been possible to specify the epidemiologic and clinical features of congenital Chagas disease. The transplacental route, as well as the role of some parasitic (genotype and parasitic charge) or host factors (capacity of maternal and fetal/neonatal immune responses, particularly the generation of CD8T cells with cytotoxic and IFN-gamma-producing capacities) in mother-to-fetus transmission of infection and/or development of congenital Chagas disease, have been also studied. Altogether, these data have allowed the development of a strategy to control T. cruzi congenital infection, which has been validated by WHO and is applied in various Latin American countries. PMID- 18429490 TI - Advances in clinical chemistry. Preface. PMID- 18429489 TI - [Epidermal stem cells]. PMID- 18429491 TI - Inhibins as diagnostic markers in human reproduction. AB - Over the past 75 years, many publications have focused on measurement of inhibin concentration and/or activity in biological samples in order to understand its role in physiology and disease. This chapter highlights the accomplishments within this area of research over the past decade including development of specific inhibin assays. Inhibin A is a marker of dominant follicle and corpus luteum activity and decreases in polycystic ovary syndrome (PCOS). Inhibin A increases in gestational diseases such as pre-eclampsia and fetal Down's syndrome, and this increase in inhibin A improves early diagnosis of both conditions. The measurement of inhibin A in women with threatened abortion provides useful information about the likelihood of pregnancy loss. Inhibin B increases markedly in women with granulosa cell tumor and appears closely related to gametogenesis in men, that is, reflecting Sertoli cell activity. On the contrary, Inhibin B decreases in women with declining ovarian function and correlates with female response to ovulation induction. This review evaluates the biochemical significance ofinhibins including their use in clinical practice. PMID- 18429492 TI - uPA and PAI-1 in breast cancer: review of their clinical utility and current validation in the prospective NNBC-3 trial. AB - The plasminogen activator system is a complex system with multiple interactions and members participating in fibrinolysis, cell migration, angiogenesis, wound healing, embryogenesis, tumor cell dissemination, and metastasis in a variety of solid tumors. Increased levels of uPA and/or PAI-1 in primary tumor tissues of breast cancer patients correlate with tumor aggressiveness and poor clinical outcome. Patients with high tumor tissue antigen content of uPA and/or PAI-1 have a worse probability of disease-free and overall survival than patients with low levels of both of the biomarkers, serving as prognostic markers. The clinical utility of uPA and PAI-1 has been proven on the highest level of evidence (LOE I). Next to being clinically useful prognostic factors allowing estimates of the course of disease in early breast cancer, uPA and PAI-1 may also serve as predictive factors predicting response to systemic therapy. Node-negative primary breast cancer patients with high uPA/PAI-1 levels benefit significantly from adjuvant chemotherapy. The aim of the ongoing NNBC-3 trial is to determine the benefits of a sequential anthracycline-docetaxel regimen in high-risk node negative breast cancer patients compared to the current standard of anthracycline based chemotherapy. At present, uPA and PAI-1 provide the unique opportunity to allow validated and clinically relevant risk assessment of breast cancer patients, over and above that provided by established risk factors. Therefore, in the evidence-based, annually updated AGO guidelines for breast cancer management, the German Working Group for Gynecological Oncology (AGO) has recommended both biomarkers as risk-group-classification markers for routine clinical decision making in node-negative breast cancer, next to established clinical and histomorphological factors. PMID- 18429493 TI - Advances in multiple analyte profiling. AB - The advent of multiparameter technology has been driven by the need to understand the complexity in biological systems. It has spawned two main branches, one in the arena of high-content measurements, primarily in microscopy and flow cytometry where it has become commonplace to analyze multiple fluorescence signatures arising from multiple excitation sources and multiple emission wavelengths. Microscopy is augmented by topographical content that identifies the source location of the signature. The other branch involves multiplex technology. Here, the intent is to measure multiple analytes simultaneously. A key feature of multiplexing is an address system for the individual analytes. In planar arrays the address system is spatial, in which affinity reactions occur at defined locations. In suspension arrays, the address is encoded as a fluorescent signature in the particle assigned to a specific reaction or analyte. Several hybrid systems have also been developed for multiplexing. In the commercial regime, the most widespread applications of multiplexing are currently in the areas of genome and biomarker analysis. Planar chips with fixed arrays are now available to probe the entire genome at the level of message expression and large segments of the genome at the level of single nucleotide polymorphism (SNP). In contrast, suspension arrays provide the potential for probing segments of the genome in a customized way, using capture tags that locate specific oligonucleotide sequences to specific array elements. PMID- 18429494 TI - Immune monitoring of clinical trials with biotherapies. AB - Immune monitoring of biotherapy clinical trials has undergone a considerable change in recent years. Technical advances together with new insights into molecular immunology have ushered a new genre of assays into immune monitoring. Single-cell assays, multiplex profiling, and signaling molecule detection have replaced formerly used bulk assays, such as proliferation or cytotoxicity. The emphasis on immune cell functions and quantitation of antigen-specific T cells has been playing a major role in attempts to establish correlations between therapy-induced alterations in immune responses and clinical endpoints. However, this has been an elusive goal to achieve, and there is a special need for improving the quality of serial monitoring to ensure that it adequately and reliably measures changes induced by administered biotherapy. In this respect, monitoring performed in specialized reference laboratories operating as good laboratory practice (GLP) facilities and strengthening of interactions between the clinical investigator, the clinical immunologist, and the biostatistician are crucial for successful use of immune monitoring in clinical studies. PMID- 18429495 TI - Dietary modification of brain function: effects on neuroendocrine and psychological determinants of mental health- and stress-related disorders. AB - Stress is associated with both psychological and biological adaptation. Chronic stress, however, impairs adaptation, and may finally lead to illness, in part through unhealthy changes in nutritional behavior. This chapter shows how physiological and psychological stress responses are affected by different food ingredients, and how stress affects health behavior, for example food choice. It becomes obvious that nutrition is closely linked to food choice and that food ingredients affect a broad range of neuroendocrine and related psychological processes, which regulate adaptation to chronic stress. Thus, dietary modification may become a valuable tool to modify the susceptibility to stress and stress-related disorders. PMID- 18429496 TI - Menopause, estrogen, and gonadotropins in Alzheimer's disease. AB - For decades, Alzheimer's disease (AD) has been linked to aging, gender, and menopause. Not surprisingly, this led most investigators to focus on the role of estrogen. While undoubtedly important, estrogen is unlikely the key determinant of disease pathogenesis. Rather, it appears that estrogen may work in conjunction with a novel determinant of disease pathogenesis, namely gonadotropins. The fact that gonadotropins, specifically luteinizing hormone, play a pivotal role in disease is apparent from significant etiological, epidemiological, and pathological evidences. Moreover, targeting gonadotropins appears to have beneficial actions as a therapeutic regimen. PMID- 18429497 TI - Immunosuppression routed via the kynurenine pathway: a biochemical and pathophysiologic approach. AB - In the past years, it has been shown that kynurenines pathway is a regulator of both the innate and the adaptive immune responses. Particularly, the initial enzyme of this pathway, indoleamine 2,3-dioxygenase (IDO), is implicated in maintaining tolerance during pregnancy, and also can be expressed in tumors to avoid the immune attack. In this chapter, we will describe how the kynurenine pathway affects the immune system with important implications both in physiology and in pathology. The incorrect activation or blockade suppressive properties of the kynurenine pathway are also implicated in a number of other diseases such as AIDS or autoimmune diseases. PMID- 18429498 TI - Pathophysiology of tumor-associated macrophages. AB - The macrophage is an important component of the human immune defense mechanism. Cancer cells secrete a variety of chemoattractants that attract macrophages and cause them to accumulate in the tumor tissue, wherein the macrophage becomes a tumor-associated macrophage (TAM). Recent evidence has shown that the function of tumor stromal TAMs can be modified by cancer cells and the factors they secrete. TAMs are directed toward stimulating tumor growth and progression and thus have protumorigenesis activity. However, there is also limited evidence that TAMs still play an important role in the killing and destruction of cancer cells, inhibit cancer metastasis, and have antitumor activity. Whether TAMs show protumorigenesis or antitumor activity depends on interaction with cancer cells, other stromal cells, and the tumor microenvironment. Gene expression profiles of TAMs, cancer cells, and other stromal cells are altered by cell-cell interactions. This phenomenon results in positive or negative regulation of angiogenesis, tumor cell proliferation, apoptosis, cancer cell migration and invasion, and the secretion of a variety of cytokines or factors. Whether TAMs have a positive or negative effect also depends on the macrophage activation state, the status of tumor development, and the anatomic locus of macrophage infiltration. Understanding of the mechanisms that regulate TAM function is essential in designing therapies to promote antitumor activity in humans. Although limited evidence from both animal and human studies indicates a potential role for TAMs in cancer treatment, the clinical usefulness of these therapies require further studies. PMID- 18429499 TI - [Veterinary medical drugs is a concern to all of us!]. PMID- 18429500 TI - [The seroprevalence of Actinobacillus pleuropneumoniae in Swiss pig breeding herds--a study with the ApxIV ELISA]. AB - At the end of the national eradication program for Enzootic Pneumonia (EP) and Porcine Actinobacillosis (APP) in Switzerland (2003), A. pleuropneumoniae serotype 2 is considered to have been eradicated. There is no current information about the distribution of the other serotypes available. The ApxIV ELISA detects antibodies against all serotypes of A. pleuropneumoniae, without cross-reaction with other bacterial species. The aim of this study was to achieve actual data concerning the seroprevalence of A. pleuropneumoniae in breeding-herds and to validate the ApxIV ELISA under field conditions, especially for the diagnosis of latently infected breeding-herds without clinical signs, and to achieve more information about the role of herd book farms for the spread of the infectious agent. A total of 2068 serum samples from 96 pig herds in Switzerland were examinated. Over half of the examinated herd book farms showed positive results in this ELISA. 93% of the breeding herds were positive. On single animal level sensitivity was 96% and specifity 100%. Herd sensitivity ranged between 67% and 99%. Herd specifity was 100%. The results show that the ApxIV ELISA is a valuable tool for the detection of latently infected herds. PMID- 18429501 TI - [Do cows drink calcium?]. AB - The objective of this study was to investigate how well cows drink the Propeller calcium drink, and it's effect on blood calcium concentration. Drinking was tested in 120 cows right after calving, before cows drank anything else. 60 cows each were offered 20 liters of Propeller calcium drink or 20 liters of water. Cows drank the Propeller as good as water. 72% of all cows drank all 20 liters, 18% drank on average 8.2 liters and 10% drank less than 1 liter. Blood calcium concentration was studied in 16 cows right after calving. Eight cows each were offered 20 liters of Propeller calcium drink or no calcium drink. Blood calcium significantly increased ten minutes after Propeller intake and stayed significantly elevated for 24 hours. Without calcium drink blood calcium levels decreased significantly. Advantages of the new Propeller calcium drink over calcium gels or boli could be that cows now drink calcium themselves and that the Propeller increases blood calcium concentration rapidly and long lasting. PMID- 18429502 TI - Comparison between an intradermal skin test and allergen-specific IgE-ELISA for canine atopic dermatitis. AB - The aim of this study was to compare the results of an intradermal skin test (IDST) with those of an allergen-specific IgE-ELISA in 210 dogs with atopic dermatitis. All the dogs had a clinical diagnosis of atopic dermatitis and underwent an IDST. The sera of all dogs were analysed for allergen-specific IgE by ELISA using the monoclonal antibody D9 against dog IgE. IDST was used as the standard assay. In both methods, the following antigens provided a positive test result: Dermatophagoides farinae, Acarus siro, Tyrophagus putrescentiae, ragweed, mugwort and Lepidoglyphus destructor. ELISA had an overall sensitivity of 82.4% and an overall specificity of 93.8%. The overall accuracy of the ELISA was 91.3%. The evaluated monoclonal D9 ELISA was found to be a reliable tool for the diagnosis of those allergens that cause clinical atopy, and can be recommended for use in dogs when immunotherapy is a therapeutic option. PMID- 18429504 TI - [The monitoring program of bluetongue disease in Switzerland]. PMID- 18429503 TI - Intracranial meningiomas associated with cervical syringohydromyelia in a cat. AB - A 13-year-old, female neutered, domestic shorthair indoor cat was referred to our hospital for treatment of multiple meningiomas. A slight generalized ataxia was seen, proprioception was severely decreased on all four limbs, and menace reaction was bilaterally reduced. Pre- and postoperatively MRI examination were performed. Three supratentorial extra-axial lesions were imaged. The fourth mass was localized infratentorial extra-axial overlying the left cerebellar hemisphere. The caudoventral cerebellum had herniated caudally, approximately one cm through the foramen magnum. Cervical syringohydromyelia was found as coincidental finding. Multiple craniotomies, centered over the meningiomas were performed. Postoperative outcome two years after the surgery is excellent. The authors also reviewed the veterinary and human literature about intracranial tumors associated syringohydromyelia. Generally, the treatment of syringohydromyelia should be targeted at the pathological process, which causes the obstruction of the cerebrospinal fluid flow, and leads to syringohydromyelia formation. PMID- 18429505 TI - [Legal case: treatment cost in found animals]. PMID- 18429506 TI - Technology: stemming the tide. PMID- 18429507 TI - Marion's message. The Death of queen Jane... PMID- 18429508 TI - Helping women avoid unnecessary interventions. PMID- 18429509 TI - Choose & lose: promoting cesareans and other invasive interventions. PMID- 18429510 TI - Just a plain birth. PMID- 18429511 TI - Technology & fear. PMID- 18429512 TI - Childbirth in the ancient Roman world: the origins of midwifery. PMID- 18429514 TI - Midwives honor the sensuality of pregnancy and birth. PMID- 18429513 TI - The future of obstetric technology. PMID- 18429515 TI - Episiotomy, hospital birth and cesarean section: technology gone haywire--what is the sutured tear rate at first births supposed to be? AB - One hundred percent of woman having a first vaginal birth from 1940-1990 had an episiotomy. It is still used for first births at a rate of 50-60% in many places. Perineal outcomes on first births are critical because the biggest risk factor for needing suturing on subsequent births is a previous episiotomy. No scientific evidence has demonstrated improved outcomes with episiotomy. This paper documents a rate of 99% intact perineums, 1% sutured perineums, in a group of 80 primipara in their early 20s at attended homebirths, average birth weight 3150 gm. Primipara women in their late-20s with 3400 gm babies experienced a 28% sutured tear rate at planned home-births. This suggests that homebirth with a motivated attendant, young age and birth weight of 3150 gm can almost always deliver vaginally without perineal damage. Episiotomy, hospital birth for healthy pregnancies and elective cesarean surgery are commonly practiced, dangerous, out of-date medical routines unsupported by research. PMID- 18429516 TI - Face to face: a midwife's birth story. PMID- 18429517 TI - Cervicometry: what all women need to know. PMID- 18429518 TI - Blogging to boost your birthing business. PMID- 18429519 TI - Listening to survivor moms. PMID- 18429520 TI - Institutionalizing midwifery: US and UK style. PMID- 18429521 TI - Birth rape: another midwife's story. PMID- 18429522 TI - What can you do to make changes with care today? PMID- 18429523 TI - Ecology of birth. PMID- 18429524 TI - Report on Uganda. PMID- 18429525 TI - Midwifery & birthing: women in Peru. PMID- 18429526 TI - Report from the field: midwifery in Hungary--an evolution. PMID- 18429527 TI - Amicus Maternity Center: part II. PMID- 18429528 TI - [How is it going with MS patients in Germany?]. PMID- 18429529 TI - [Laboratory parameters]. AB - Numerous physiologic parameters can be analyzed in blood and urine. In many pharmacies, pharmacy support staff perform diagnostic analytic tests like blood glucose and cholesterol tests on a daily basis. Occasionally they perform standard liver function tests as well. Frequently customers attend the pharmacy and ask for the significance of particular laboratory test results. These are enough reasons to discuss laboratory values, the associated medical consequences and the factors which possibly affect the test results in greater detail. PMID- 18429530 TI - [Complementary and alternative therapies for multiple sclerosis]. PMID- 18429531 TI - [Thujone-attributable effects of absinthe are only an urban legend--toxicology uncovers alcohol as real cause of absinthism]. AB - In the discussion about thujone as possible toxic constituent of the wormwood containing alcoholic beverage absinthe, the dose-response-relationship is frequently ignored. The effects of absinthe are very often attributed only to thujone, an association that is not scientifically proven. Especially the alleged psychotropic effects of thujone are scientifically unproven. However, the question about thujone effects in absinthe is irrelevant, because thujone is contained in both modern commercial absinthes and historic pre-ban products in such low amounts that a pharmacological effect can be excluded per se. The effects of the spirit that are summarized under the term absinthism observed in late 19th century's France, can be explained by chronic alcohol misuse and dependence alone according to today's standards of knowledge. Especially from the perspective of youth and public health protection, an ambiguous and biased reporting about absinthe should be avoided. For example, the alleged antagonistic effects of thujone on the action of ethanol might lead to a trivialization of alcohol-related harms. Scientifically unproven speculations about the influence of certain drinking rituals of absinthe on its toxicity must be rebutted. A return to more evidence and less conjecture in the reporting about absinthe would be desirable. PMID- 18429532 TI - [Prescription of rimonabant in the early stage of pregnancy?]. AB - Rimonabant should not be prescribed in pregnancy because no sufficient data on the teratogenic/ embryotoxic risk are available. A preventive termination of pregnancy is not indicated. Women in childbearing age should take only low-risk drugs. Otherwise dedicated information, in doubt pregnancy test, and strict contraception is required. PMID- 18429533 TI - 'Doctor:' for physicians only? PMID- 18429534 TI - Publication: a mark of professsionalism. PMID- 18429535 TI - Case study: obstructive sleep apnea. AB - Obstructive sleep apnea (OSA) is a major health problem that often goes undiagnosed. A case study is utilized to illustrate recognition, diagnosis, and treatment of individuals with OSA. Prevalence, risk factors, and health and safety issues also are discussed. PMID- 18429536 TI - Evaluation of a device to facilitate female urethral catheterization. AB - Urethral catheterization is a skilled procedure that nurses in hospital settings perform routinely. The opening of the female urethra is located within the vulvar vestibule, making insertion of urinary catheters into females a greater technical challenge than in males. Researchers evaluated whether a new device might decrease the time required for catheter insertion, increase the likelihood of inserting the catheter on the first attempt (improved accuracy), and reduce patient discomfort. Comments about the device from both patients and nurses also are reported. PMID- 18429537 TI - Tackling sundowning in a patient with Alzheimer's disease. AB - The early evening exacerbation of behavioral symptoms in sundowning has been recognized by medical professionals as a challenge. The circadian, hormonal, physiological, and environmental correlations with sundowning behaviors are described. PMID- 18429538 TI - Hypoglycemia in hospitalized adults. AB - Hypoglycemia is a commonly occurring iatrogenic event in hospitalized patients. Nurses play an important role in the detection, treatment, and prevention of hypoglycemia. The literature regarding hypoglycemia in hospitalized adults and implications for nursing practice is reviewed. PMID- 18429539 TI - Nurse-physician collaboration on medical-surgical units. AB - Interdisciplinary collaboration is viewed as a critical factor in delivering quality patient care. The purpose of this study was to describe nurse-physician perceptions of collaboration relationship on general medical-surgical units. PMID- 18429540 TI - Research considerations: power analysis and effect size. PMID- 18429541 TI - Cervical cancer: an overview with suggested practice and policy goals. PMID- 18429542 TI - Herb-drug interactions. PMID- 18429543 TI - Nurse knowledge, skills, and attitudes related to evidence-based practice: before and after organizational supports. PMID- 18429544 TI - Staff engagement: it starts with the leader. AB - * Nursing cannot grow stronger unless we can recruit and sustain a cadre of engaged, spirited, and involved leaders. * The body is designed to work most effectively in a series of 90-minute increments with a recovery time in between. * The science of leadership/management can be seen as a game of chess where the pieces can move in all directions all over the board based on the uniqueness of each player. * Creating an optimistic culture where people feel they have hope and freedom to grow and mature will create a sense of engagement. PMID- 18429545 TI - Saddened by scrofulousness. PMID- 18429546 TI - Labs are vital. PMID- 18429547 TI - Looming threat to QC in physicians' offices. PMID- 18429548 TI - What is on the molecular diagnostics horizon? Part 5b--a brief history of medical diagnosis and the birth of the clinical laboratory. PMID- 18429549 TI - Future of molecular diagnostics and cancer. PMID- 18429550 TI - LEAN deploys at Centrex Clinical Labs. PMID- 18429551 TI - Improve testing turnaround by looking beyond the lab. PMID- 18429552 TI - Endangered species vanishing from medical labs. PMID- 18429553 TI - OSHA enforcement activity on BPS: an update. PMID- 18429554 TI - HIPAA guides answering services. PMID- 18429555 TI - Creating a mutually beneficial approach to CBD project. PMID- 18429556 TI - Patient's page. Meth mouth--a brief overview. PMID- 18429557 TI - ODA/Schools for Healthy Lifestyles (SHL) "Adopt-A-Dentist" program. PMID- 18429558 TI - 2007 costs and coverage of antiretrovirals under Medicare Part D for people with HIV/AIDS living in North Carolina. AB - BACKGROUND: Effective January 1, 2006 Medicare Part D became a new source of prescription drug coverage for people with HIV/AIDS in the United States. The implementation of Part D has affected access to antiretrovirals for people with HIV/AIDS. In North Carolina, access can be difficult because of the state's struggling safety net programs and the growing HIV-infected populations among Blacks and in poor rural counties. This analysis examines Medicare Part D antiretroviral coverage in 2007 for beneficiaries with HIV/AIDS in North Carolina, particularly those who did not qualify as dual eligibles or for a full low-income subsidy. METHODS: Data describing program coverage were obtained from the Web site www.medicare.gov and descriptive analyses were performed to assess changes in antiretroviral coverage in Part D prescription drug plans in North Carolina. RESULTS: Most of the 26 antiretrovirals are covered in some way by 76 North Carolina prescription drug plans. There may be variability in coverage however associated with (a) antiretroviral classification within formularies; (b) drug premiums; (c) whether premiums can be waived; (d) annual deductibles; and (e) whether coverage is provided in the "doughnut hole." LIMITATIONS: The data may not reflect actual patterns of drug use and realized access to the drugs. The findings are limited to antiretroviral coverage in North Carolina's Part D offerings but could be generalized to other states with similar prescription drug plan costs and coverage. CONCLUSION: These concerns continue to pose significant challenges to accessing antiretrovirals for Part D beneficiaries with HIV/AIDS in North Carolina. Variability demonstrated within prescription drug plans will continue, and beneficiaries with HIV/AIDS who do not qualify as dual eligibles or for low-income subsidies will need to evaluate these issues when selecting a prescription drug plan in future enrollment periods. PMID- 18429559 TI - Tuberculosis knowledge, attitudes, and beliefs among North Carolinians at increased risk of infection. AB - OBJECTIVE: The purpose of this study was to capture and describe knowledge, attitudes, and beliefs about tuberculosis (TB) among persons at high risk for TB infection. METHODS: We conducted 11 focus groups in 3 different populations at high risk for TB infection: Spanish-speaking immigrants, homeless shelter residents, and persons attending a drug/alcohol rehabilitation center. A standardized list of open-ended questions was used to guide discussion. Using grounded theory, transcripts of the focus group sessions were reviewed by 4 independent reviewers to identify emergent themes. FINDINGS: Participants (N = 52) generally understood that TB is an infectious disease that frequently affects the lungs and can be fatal if untreated. They also knew that a skin test can be used to diagnose TB. However, participants frequently had incorrect beliefs regarding the cause, transmission, and treatment of TB. Many participants thought that TB is transmitted in the same fashion as other infectious diseases such as human immunodeficiency virus or aquired immune deficiency syndrome (HIV/AIDS). A general sentiment of fear and aversion toward persons ill with TB was expressed. LIMITATIONS: Focus groups were a convenience sample and subjects were not necessarily representative of the underlying populations. CONCLUSIONS: Tuberculosis knowledge among high-risk populations is suboptimal, and false beliefs regarding transmission and treatment were common in this study. Knowledge regarding transmission of other infectious diseases such as HIV/AIDS was frequently translated into incorrect knowledge regarding TB. Stigma continues to be a barrier to TB diagnosis and treatment. PMID- 18429560 TI - Policy forum: taking care of those who serve. PMID- 18429561 TI - Serving the health needs of our military and veterans. PMID- 18429562 TI - The Veterans Affairs health care system in North Carolina. PMID- 18429563 TI - The epidemiology of health problems in returning Operation Iraqi Freedom and Operation Enduring Freedom veterans. A national and North Carolina-based summary. PMID- 18429564 TI - Veterans Affairs Research and Development: using science to improve health care for veterans. AB - VA has a rich tradition in supporting research in areas that span basic science to health system implementation. Its unique success is tied to the fact that researchers are focused on issues that arise from a unique population--our nation's veterans. Moreover, because VA is the largest integrated health system in the country and because the health system must manage an annual budget, there is a keen interest among VA health administrators to apply research that enhances quality and efficiency of care. Furthermore, because these findings overlap with the general population, VA Research & Development programs can be applicable on a much broader scale. PMID- 18429565 TI - Identifying and treating post deployment mental health problems among new combat veterans. PMID- 18429566 TI - Detecting traumatic brain injury among veterans of Operations Enduring and Iraqi Freedom. PMID- 18429567 TI - Veterans Health Administration primary care-mental health integration initiative. PMID- 18429568 TI - Integrating primary care and mental health in the Army. PMID- 18429569 TI - The Citizen Soldier Support Program: building bridges between communities and military families. PMID- 18429570 TI - Military families: opportunities and challenges. PMID- 18429571 TI - Durham Veterans Affairs Medical Center. PMID- 18429572 TI - Mental health professionals in North Carolina. PMID- 18429573 TI - Courage and character, leaders and legends: an interview with Richard Lower, MD by Linda Ohler. PMID- 18429574 TI - Subcapsular hematoma evacuation as a method of evaluating injured kidneys for transplant. AB - BACKGROUND: Approximately one-third of organ donors in the United States are trauma victims. In general, kidneys with large subcapsular hematomas are not used for transplant because of the possibility of significant parenchymal injury. A large subcapsular renal hematoma may cause scarring resulting in renal parenchymal compression and development of the Page syndrome. OBJECTIVE: To elucidate a successful method of evaluating kidneys subject to trauma, while also possibly preventing further damage and improving their function. DESIGN: Data were collected from the donor kidney pool of the New York Organ Donor Network from January 2006 through July 2007. Four kidneys during that period were determined to have significant subcapsular hematomas. Surgical intervention was undertaken and outcomes after transplantation were reviewed. MAIN OUTCOME MEASURES: Four of the kidneys underwent a surgical procedure to drain the subcapsular hematoma allowing assessment of the underlying renal parenchyma. All 4 of these kidneys were deemed transplantable. After transplantation, 3 of the 4 kidneys had immediate function and did not require dialysis. The remaining kidney was removed as a result of primary nonfunction. CONCLUSION: The described surgical intervention allows the transplant surgeon to accurately assess the extent of damage to a traumatized kidney while possibly preventing further damage to the kidney. PMID- 18429575 TI - Improving equity of access to deceased donor kidneys in New Zealand. AB - Before 1999, deceased donor kidneys in New Zealand were allocated by local nephrologists on the basis of local needs and factors thought to be beneficial to graft outcome-in particular, matching tissue type. Waiting times were much longer for patients whose tissue type was different from that of our mainly white population of deceased donors. A National Kidney Allocation System was developed that improved equity of access for our unique population base in New Zealand, taking into account not only tissue-type matching but time spent waiting. PMID- 18429576 TI - When the donor says yes and the family says no. AB - Signed donor cards clearly demonstrate the donor's intention to donate organs after death. In many states, this donation cannot be rescinded by the next of kin, and organs can be recovered from the donor even if the family objects. The family usually does not object if the donor has signed an organ donor card, especially if the donor had discussed the issue with the family. In some situations, however, the family objects to donation despite the signed organ donor card. If the organ procurement organization pursues donation, adverse publicity and even legal action are possible. It can be a challenge for organ procurement personnel to deal with families who object to donation in the face of a signed organ donor card in a manner that will lead to successful organ recovery without adverse consequences. This article describes 4 cases where the donor had a signed organ donor card but the family initially objected to donation. Ultimately organs were recovered from 3 of these donors. PMID- 18429577 TI - Timely referral of potential organ donors. PMID- 18429578 TI - Factitious wound infections in an altruistic living liver donor. AB - We report a case of recurrent factitious wound infections in an altruistic living liver donor. Costs for the infections after donation were billed to the recipient, creating a threat to the recipient's lifetime benefits. This case illustrates the importance of obtaining previous medical records on living donors. PMID- 18429579 TI - Patients' willingness to talk to others about living kidney donation. AB - BACKGROUND: Living donor kidney transplantation has several advantages for patients with end-stage renal disease. However, many patients are reluctant to pursue this treatment option, preferring instead to wait for a deceased donor organ. OBJECTIVE: To examine predictors of patients' willingness to talk to others about living kidney donation. METHODS: One hundred thirty-two adult patients awaiting kidney transplantation who were enrolled in a randomized trial examining the effectiveness of education on rates of live donor kidney transplantation completed a baseline rating of their willingness to talk to others about living kidney donation. Also, patients completed measures of knowledge and concerns about living donation and a rating of perceived health. RESULTS: Slightly more than half the patients (56.1%) had low willingness to talk to others about living donation. The following variables were associated with higher willingness to talk to others: white race (odds ratio, 3.31; confidence interval, 1.7-7.4), college education (odds ratio, 3.43, confidence interval, 2.0 5.6), fewer concerns about living donor kidney transplantation (odds ratio, 0.31; confidence interval, 0.2-0.6), and less favorable perceptions of their current health status (odds ratio, 4.31; confidence interval, 2.6-7.6). CONCLUSION: White race, more education, less concern about living donor kidney transplantation, and poorer perceived health are associated with greater willingness to talk to others about living kidney donation. These findings have important implications for educating patients about living donor kidney transplantation. PMID- 18429580 TI - An international perspective: job satisfaction among transplant nurses. AB - CONTEXT: The high demand for transplant nurses across the world leads us to examine job design and job satisfaction because job satisfaction is linked to better outcomes for patients. OBJECTIVE: To describe international transplant nurses' perspectives of job design and job satisfaction by using Herzberg's theory of motivation. METHODS: Descriptive, correlational design. An electronic version of the Job Design and Job Satisfaction survey was mailed to all members of the International Transplant Nurses Society. RESULTS: A total of 331 members of the International Transplant Nurses Society responded to the survey. The mean age of respondents was 44.12 years, they had worked a mean of 19.12 years in nursing and 10.22 years in transplantation, and 50.6% of respondents were transplant nurse coordinators. Respondents were very satisfied overall with their jobs; they perceived that transplant nursing requires a high level of nonrepetitive, complex skills, autonomy in personal initiative and judgment, cooperation and collaboration with others, and that the job allows for completion of the work. Respondents were satisfied with pay, fringe benefits, and supervision. The feeling that the job could positively and significantly affect others was very strong. CONCLUSIONS: Results of this study provide empirical evidence supporting the perceived benefits and challenges of working in transplantation and support Herzberg's theory that motivators leading to job satisfaction include achievement, recognition, the work itself, responsibility, and advancement. Transplant nursing includes many of these motivators and desirable characteristics, including autonomy and working with a multidisciplinary team on a clear, patient-centered goal. PMID- 18429581 TI - Consideration of psychosocial factors in the evaluation of living donors. AB - Results of donor outcome studies indicate that most living donors report a positive psychosocial response to donation. However, negative psychosocial outcomes have also been reported. Evaluation guidelines have been proposed, although a standardized evaluation specific to living donors is not yet available. In an effort to determine what psychosocial factors should be considered in a comprehensive evaluation of living donors, an extensive literature review was undertaken that was focused on previously proposed guidelines for the psychosocial evaluation of living donors, research on outcomes among living donors, and other relevant psychosocial data. PMID- 18429582 TI - Further exploration: maturity and adherence in adolescent and young adult heart transplant recipients. AB - CONTEXT: Pediatric transplant clinicians note high rates of nonadherence with medications, appointment keeping, and laboratory tests and high rates of engagement in high-risk behaviors among older adolescents and young adult recipients. The caregivers also report symptoms of identity confusion, social immaturity, and failure to appreciate consequences of risky behavior among recipients. OBJECTIVE: To build on an earlier study that identified developmental characteristics that distinguish poor from good adherers. DESIGN: Qualitative methods were used to explore key themes identified in the first study with heart transplant recipients and their significant others. RESULTS: This research confirmed the themes of developmental maturity suggested by the first study: good adherers were able to integrate the transplant into sense of self, become independent adults, and achieve normalcy; poor adherers continued to "push away" the transplant experience and worried that they would never be normal. The good adherers, their parents, and friends had views of relationships that were congruent, acknowledging difficulties, discussing them, and moving toward mutual satisfaction. The poor adherers, their parents, and friends expressed incongruent views of the relationships, avoiding discussion of problems and idealizing relationships. DISCUSSION: Strategies for clinicians, family, and friends to increase maturity and independence among older adolescent and young adult heart transplant recipients are described. PMID- 18429583 TI - Optimal transplant education for recipients to increase pursuit of living donation. AB - CONTEXT: Inadequate transplant education may stop kidney patients from beginning or completing evaluation or limit recipients from considering living donation. OBJECTIVE: To learn about recipients' decision making about living donation and preferred transplant education resources. DESIGN: Retrospective, cross-sectional survey. PATIENTS: 304 kidney recipients. MAIN OUTCOME MEASURES: Living donation comfort, concerns, education preferences. RESULTS: Recipients spent 10 median hours learning about transplant, primarily by speaking to medical staff (2-3 hours) and reading transplant brochures (0-1 hour). Twelve percent had not received any education before coming to the transplant center. At least 75% wanted education discussing the evaluation, surgery, and medical tests required of recipients and donors, as well as common transplant-related fears. Recipients who received living donor transplants were more interested in information about donors' evaluation (P < .001), surgery (P < .001), medical tests (P < .001), and donation concerns (P = .004) than were other recipients. Recipients who had living donors evaluated were more comfortable accepting family members or friends who volunteered rather than asking potential donors because of concerns about pressuring donors (85%), harming their health (83%), or causing them pain or inconvenience (76%). Besides providing accurate medical information, education that addresses recipients' fears about transplantation, explains living donors' donation experiences, and teaches patients how to pursue living donation may increase recipients' pursuit of living donation. PMID- 18429584 TI - Milestones in transplantation. PMID- 18429585 TI - Virginia Tech's implications. PMID- 18429586 TI - Finance must be everyone's business. PMID- 18429587 TI - What's in the pipeline. PMID- 18429588 TI - Protecting ownership of patient information. PMID- 18429589 TI - Making the recovery model real. PMID- 18429590 TI - Balancing access and security. PMID- 18429591 TI - APRNs on the GO. PMID- 18429592 TI - Process versus product. PMID- 18429593 TI - Lassoing collaboration to spur change. PMID- 18429594 TI - Medication is not a 'silver bullet'. PMID- 18429595 TI - New substrates and reversible inhibitors of cholinesterases based on esters and thioesters of noncholine cyclic ammonium alcohols. PMID- 18429596 TI - Bioactive ceramic tantalum-doped films for implants. PMID- 18429597 TI - Catalytic activity of nanodiamond particles in organic reactions. PMID- 18429598 TI - Substrate specificity of the anthrax lethal factor. PMID- 18429599 TI - Transformation of hydrocarbons into components of archaic chemoautotrophic CO2 fixation cycle. PMID- 18429600 TI - The effect of silver ions on copper metabolism and expression of genes encoding copper transport proteins in rat liver. PMID- 18429601 TI - Amyloid sarcomeric proteins of the titin family. PMID- 18429602 TI - Ganglioside GM1-binding sites in interleukin-4: a photoaffinity labeling study. PMID- 18429603 TI - Ecological-genetic organization of plant quantitative traits and mapping of the loci determining agronomically important traits in soft wheat. PMID- 18429604 TI - A targeted transport of 125I-labeled monoclonal antibodies to target proteins in experimental glioma focus. PMID- 18429605 TI - Carnosine protects erythrocytes from the oxidative stress caused by homocysteic acid. PMID- 18429606 TI - Modeling of the relationships between the structure of O-phosphorylated oximes and their anticholinesterase activity and selectivity using molecular field topology analysis (MFTA). PMID- 18429607 TI - Novel fluorescent core-shell nanocontainers for cell membrane transport. AB - The synthesis and characterization of novel core-shell macromolecules consisting of a fluorescent perylene-3,4,9,10-tetracarboxdiimide chromophore in the center surrounded by a hydrophobic polyphenylene shell as a first and a flexible hydrophilic polymer shell as a second layer was presented. Following this strategy, several macromolecules bearing varying polymer chain lengths, different polymer shell densities, and increasing numbers of positive and negative charges were achieved. Because all of these macromolecules reveal a good water solubility, their ability to cross cellular membranes was investigated. In this way, a qualitative relationship between the molecular architecture of these macromolecules and the biological response was established. PMID- 18429608 TI - Free radical formation in supercritical CO2, using muonium as a probe and implication for H atom reaction with ethene. AB - This report presents the first observation of an alkyl radical in supercritical CO(2) by any magnetic resonance technique. Muoniated ethyl radical has been detected in muon-irradiated supercritical CO(2) solutions. In the presence of a low concentration of ethene in supercritical CO(2), it is found that the addition of muonium to ethene is the only reaction channel, and that the yield of this process is enhanced compared to conventional solvents. The temperature dependence of the hyperfine coupling constants of the ethyl radical suggests that at a density of 0.3 g/cm(3) both the rotational motion of the methyl group and the electronic structure of the radical are similar to those in the gas phase, and therefore that the local environment around the ethyl radical is similar to the gas phase under these conditions. At higher densities, however, there is a remarkable and unexpected density dependence of the hyperfine coupling constant of the ethyl radical, which has never been observed in any environment. In this regime, the density dependence suggests that supercritical CO(2) has a significant effect on the electronic structure of the free radical. Thus, changing the density of CO(2) offers a possible means of tuning the radical reactivity. In addition, at a density of close to 0.4 g/cm(3), CO(2) molecules cluster around the ethyl radical, and this increases the local density around the ethyl radical by a factor of ~1.5. PMID- 18429609 TI - From the selective serotonin transporter inhibitor citalopram to the selective norepinephrine transporter inhibitor talopram: synthesis and structure-activity relationship studies. AB - Citalopram and talopram are structurally closely related, but they have very distinct pharmacological profiles as selective inhibitors of the serotonin and norepinephrine transporters, respectively. A systematic structure-activity relationship study was performed, in which each of the four positions distinguishing the two compounds were varied. The inhibitory potencies of the resulting 16 compounds were tested at both serotonin and norepinephrine transporters. This showed that particularly two of the four positions are determinants for the biological activity. PMID- 18429610 TI - Spiro[(dihydropyrazin-2,5-dione)-6,3'-(2',3'-dihydrothieno[2,3-b]naphtho-4',9' dione)]-based cytotoxic agents: structure-activity relationship studies on the substituent at N4-position of the diketopiperazine domain. AB - Analogues of the previously reported potent cytotoxic spiro[(dihydropyrazine-2,5 dione)-6,3'-(2',3'-dihydrothieno[2,3-b]naphtho-4',9'-dione)] derivatives (3, 3') were prepared to explore new structural requirements at the diketopiperazine domain for the cytotoxic activity. The in vitro activity was evaluated against the MCF-7 human breast carcinoma and SW 620 human colon carcinoma cell lines. The 4-[(2-N, N-dimethyl)amino]ethyl (6i), and the 4-(2-pyrrolydin)ethyl (6l) derivatives emerged as the most potent compounds of this series, with a cytotoxic activity comparable to that of doxorubicin. These compounds, in both racemic and pure enantiomeric forms, showed also a high efficacy in cell lines resistant to doxorubicin (MCF-7/Dx) and in cell lines that were highly resistant to treatment with doxorubicin, such as HEK-293 (kidney), M-14 (melanoma), and HeLa (cervical adenocarcinoma) human cell lines. In addition, the effects on growth and cell cycle progression in CaCo-2 cell line (colon adenocarcinoma) and DNA-binding properties were investigated. PMID- 18429611 TI - Direct measurement of tertiary contact cooperativity in RNA folding. AB - All structured biological macromolecules must overcome the thermodynamic folding problem to populate a unique functional state among a vast ensemble of unfolded and alternate conformations. The exploration of cooperativity in protein folding has helped reveal and distinguish the underlying mechanistic solutions to this folding problem. Analogous dissections of RNA tertiary stability remain elusive, however, despite the central biological importance of folded RNA molecules and the potential to reveal fundamental properties of structured macromolecules via comparisons of protein and RNA folding. We report a direct quantitative measure of tertiary contact cooperativity in a folded RNA. We precisely measured the stability of an independently folding P4-P6 domain from the Tetrahymena thermophila group I intron by single molecule fluorescence resonance energy transfer (smFRET). Using wild-type and mutant RNAs, we found that cooperativity between the two tertiary contacts enhances P4-P6 stability by 3.2 +/- 0.2 kcal/mol. PMID- 18429612 TI - Palladium-catalyzed diastereo- and enantioselective Wagner-Meerwein shift: control of absolute stereochemistry in the C-C bond migration event. AB - Inducing absolute stereochemistry in Wagner-Meerwein shifts was examined in a ring expansion protocol. Initiated by generation of a pi-allylpalladium intermediate by hydropalladation of allenes, the ring expansion of allenylcyclobutanol substrates proceeded with excellent diastereo- and enantioselectivities. The results demonstrate that, during the C-C bond migration process, our chiral catalysts can control the stereochemistry of both the pi allylpalladium intermediate and the corresponding migration bond. Moreover, the stereochemical outcome of the reaction can be rationalized very well with the working model of the chiral catalyst. The method provides an efficient way to synthesize highly substituted cyclopentanones with an alpha-chiral O-tertiary center which has various synthetic applications. PMID- 18429614 TI - Fluorophore-labeled beta-lactamase as a biosensor for beta-lactam antibiotics: a study of the biosensing process. AB - The fluorescein-labeled E166C mutant of the PenPC beta-lactamase (E166Cf) represents a successful model in the construction of "switch-on" fluorescent biosensors from nonallosteric proteins (Chan P.-H. et al.; J. Am Chem. Soc., 2004, 126, 4074). This paper focuses on the study of the biosensing mechanism by which the E166Cf biosensor changes its fluorescence upon beta-lactam binding and hydrolysis. Mass spectrometric and stopped-flow fluorescence studies of E166Cf with cefuroxime, penicillin G, and 6-aminopenicillanic acid reveal that the formation of enzyme-substrate complex enhances the fluorescence of E166Cf, and the subsequent regeneration of the free enzyme restores the weak fluorescence of E166Cf. Molecular modeling studies of E166Cf with penicillin G show that the fluorescein label is likely to share a common space with the beta-lactam and thiazolidine rings of the antibiotic in the active site. This spatial clash appears to cause the fluorescein label to move from the active site to the external aqueous environment upon substrate binding and hence experience higher water exposure. Steady-state fluorescence measurements indicate that the fluorescence of E166Cf can be enhanced by 6-aminopenicillanic acid, which consists of the beta-lactam and thiazolidine rings only. Thermal denaturation experiments of the wild-type enzyme, E166C, and E166Cf reveal that the E166C mutation is likely to increase the flexibility of the Omega-loop. This "modified" structural property might compensate for the possible steric effect of the fluorescein label on substrate binding. PMID- 18429615 TI - Cipadonoid A, a novel limonoid with an unprecedented skeleton, from Cipadessa cinerasecns. AB - Cipadonoid A ( 1), a novel limonoid with an unprecedented skeleton, was isolated from the leaves of Cipadessa cinerasecns. Its structure and relative configuration were determined by spectroscopic analysis and computer modeling. 1 represents a new type of limonoid, characterized by a rearranged tetrahydropyranyl ring B incorporating usually exocyclic C-30. A possible biosynthetic pathway of 1 was also proposed. PMID- 18429616 TI - Clar-Kekule structuring in armchair carbon nanotubes. AB - Geometrical patterns on armchair nanotubes and their dependence on length (up to 10 nm) have been studied using first-principles methods. The results indicate that finite nanotubes do not show a uniform bond structure. The previous structural classification of armchair nanotubes in Clar, Kekule, and incomplete Clar types becomes unified with lengthening, not in a bond-uniform structure, as PBC models report, but into an alternated sequence of Clar and Kekule domains in all cases, with possible mechanical and electronic consequences. PMID- 18429617 TI - Gas-phase reactivity of aromatic sigma,sigma-biradicals toward dinucleoside phosphates. AB - In order to improve the understanding of the interactions of aromatic sigma,sigma biradicals with DNA, the reactivity of three isomeric sigma,sigma-biradicals toward four dinucleoside phosphates was studied in a mass spectrometer. The dinucleoside phosphates were evaporated into the mass spectrometer by using laser induced acoustic desorption (LIAD). The results demonstrate that the structure of the sigma,sigma-biradical and the base sequence of the dinucleoside phosphate can have a major influence on these reactions. PMID- 18429618 TI - Synthesis of 5-vinylideneoxazolidin-2-ones by DBU-mediated CO2-fixation reaction of 4-(benzylamino)-2-butynyl carbonates and benzoates. AB - A CO2-fixation reaction of 4-(benzylamino)-2-butynyl carbonates and benzoates, carried out in the presence of DBU, provides substituted 5-vinylideneoxazolidin-2 ones. The reaction has been successfully applied to the CO2-recycling process and fixation of atmospheric CO2. PMID- 18429619 TI - Lewis acid-Lewis base catalyzed enantioselective hetero-Diels-Alder reaction for direct access to delta-lactones. AB - A complex formed in situ from Er(OTf)3 and a simple commercially available norephedrine ligand promotes an unprecedented [4 + 2] cycloaddition of alpha,beta unsaturated acid chlorides with a broad range of aromatic and heteroaromatic aldehydes by a cooperative bifunctional Lewis acid-Lewis base catalytic mode of action providing valuable delta-lactone building blocks with excellent enantioselectivity. PMID- 18429620 TI - Facile synthesis of cis-2-Alkyl-3-trialkylsilyloxycycloalkanones via the non aldol aldol rearrangement of 2,3-epoxycycloalkanols. AB - Silyl triflate-promoted rearrangement of cis-2,3-epoxycycloalkanols A, prepared by epoxidation of the cyclic allylic alcohol and then silylation, afforded good yields ( approximately 70-75%) of the cis-2-alkyl-3-silyloxycycloalkanones B, presumably via the intermediates C and D, even with quite large alpha substituents, e.g., tert-butyl. Finally, it has been shown that the stereochemistry of the epoxy alcohol is crucial as one would expect from the mechanism. PMID- 18429621 TI - First asymmetric total syntheses of cernuane-type Lycopodium alkaloids, cernuine, and cermizine D. AB - The first total syntheses of two cernuane-type Lycopodium alkaloids, (-)-cernuine and (+)-cermizine D, were accomplished starting from (+)-citronellal. The syntheses involved organocatalytic alpha-amination to afford oxazolidinone, which is used for diastereoselective allylation, and asymmetric transfer aminoallylation followed by stereoselective construction of an aminal moiety as key steps. PMID- 18429622 TI - Red antenna states of photosystem I from Synechocystis PCC 6803. AB - Single-molecule spectroscopy at low temperatures was used to elucidate spectral properties, heterogeneities, and dynamics of the red-shifted chlorophyll a (Chl a) molecules responsible for the fluorescence from photosystem I (PSI). Emission spectra of single PSI complexes from the cyanobacterium Synechocystis PCC 6803 show zero-phonon lines (ZPLs) as well as broad intensity distributions without ZPLs. ZPLs are found most frequently on the blue side of the broad intensity distributions. The abundance of ZPLs decreases almost linearly at longer wavelengths. The distribution of ZPLs indicates the existence of at least two pools with maxima at 699 and 710 nm. The pool with the maximum at 710 nm is assigned to chlorophylls absorbing around 706 nm (C706), whereas the pool with the maximum at 699 nm (F699) can be assigned to chlorophylls absorbing at 692, 695, or 699 nm. The broad distributions dominating the red side of the spectra are made up of a low number of emitters assigned to the red-most pool C714. The properties of F699 show close relation to those of F698 in Synechococcus PCC 7002 and C708 in Thermosynechococcus elongatus. Furthermore, a high similarity is found between the C714 pool in Synechocystis PCC 6803 and C708 in Synechococcus PCC 7002 as well as C719 in T. elongatus. PMID- 18429623 TI - Dose-response relationships for cancer incidence reflect susceptibility distributions. PMID- 18429624 TI - Block copolymer micelle shuttles with tunable transfer temperatures between ionic liquids and aqueous solutions. AB - Four poly((1,2-butadiene)- block-ethylene oxide) (PB-PEO) diblock copolymers were shown to self-assemble into micelles with PB cores and PEO coronas (including spheres, cylinders, and vesicles) in the ionic liquid, 1-ethyl-3 methylimidazolium bis(trifluoromethylsulfonyl)imide ([EMIM][TFSI]). All four systems exhibited the "micelle shuttle" (He, Y.; Lodge, T. P. J. Am. Chem. Soc. 2006, 128, 12666-12667), whereby PB-PEO micelles transferred, reversibly and with preservation of micelle structure, from an aqueous phase at room temperature to a hydrophobic ionic liquid at high temperature. The micelle size (both mean and distribution) depends on whether it was initially dissolved in water or in the ionic liquid, but the initial micelle structures in the ionic liquid were shown by dynamic light scattering to be preserved during the transfer and persist essentially unchanged for months in both the ionic liquid and water. The transfer was shown to be driven by the deteriorating solvent quality of water for PEO at high temperature, while the ionic liquid remains a good solvent. The transfer temperature could be tuned by adding ionic or nonionic additives to the aqueous phase to change the solvent quality of water for PEO, and by using ionic liquids with different polarity. PMID- 18429625 TI - Time-resolved in situ small-angle X-ray scattering study of silica particle formation in nonionic water-in-oil microemulsions. AB - The formation of silica particles by the ammonia-catalyzed hydrolysis of tetraethyl orthosilicate (TEOS) in the polyoxyethylene (5) nonylphenyl ether (NP 5)/cyclohexane/water microemulsion system was investigated by time-resolved small angle X-ray scattering (SAXS). The SAXS data could be modeled as a combination of two species where one describes the silica-particle containing microemulsion droplets and the other the reverse droplets. The analysis allowed the determination of the evolution of the system of particles of silica and reverse droplets. A model of nucleation and growth of the silica particles is confirmed and the volume fraction versus time data for the silica particles is in agreement with first order kinetics with respect to TEOS concentration. Moreover to describe the long time evolution of the system, a correlation among the silica particles has been taken into account by introducing a structure factor with a local silica volume fraction eta = 0.1. This high local density is 2 orders of magnitude larger than the global silica fraction and can be explained in terms of depleting interaction. PMID- 18429626 TI - In vitro cytotoxicity, chemotactic effect, and cellular uptake of branched polypeptides with poly[L-lys] backbone by J774 murine macrophage cell line. AB - Branched polypeptides with polylysine backbone are promising candidates for selective delivery of drugs, epitopes. or reporter molecules. We reported earlier that polylysine-based polypeptides with polyanionic character were internalized by murine bone marrow derived macrophages via class A scavenger receptor. In the present studies, our investigations were extended to seven polypeptides with different amino acid composition and charge properties. We report on our findings on the concentration-dependent influence of these compounds on survival and chemotaxis of the murine macrophage-like cell line J774 and internalization properties of the polypeptides by J774 cells. Our observations indicate that the polypeptides regardless of their charge properties were essentially nontoxic and did not alter significantly the chemotaxis of J774 cells; therefore, the polypeptides suit the requirements for nontoxic and "neutral" carrier molecules. We also demonstrated that the polypeptides were internalized efficiently by J774 cells, depending on their chemical structure and charge properties. Using the scavenger receptor-ligand fucoidan as inhibitor, we established that the scavenger receptor played a role-in accordance with findings on murine bone marrow derived macrophages in the internalization only of the polyanionic polypeptides. PMID- 18429627 TI - A novel near-infrared indocyanine dye-polyethylenimine conjugate allows DNA delivery imaging in vivo. AB - Near-infrared (NIR) fluorescence light has been applied to monitor several biological events in vivo since it penetrates tissues more efficiently than visible light. Dyes exhibiting NIR fluorescence and having large Stokes shift are key elements for this promising optical imaging technology. Here, we report the synthesis of a novel conjugate between a near-infrared indocyanine dye and an organic polyamine polymer (polyethylenimine, PEI) (IR820-PEI) with high chemical stability and good optical properties. IR820-PEI absorbs at 665 nm, emits at 780 nm, and displays a large Stokes shift (115 nm). Moreover, the reported conjugate is able to bind DNA, and the delivery process can be monitored in vivo with noninvasive optical imaging techniques. These characteristics make IR820-PEI one of the most effective and versatile indocyanine dye polymeric-conjugate reported so far. PMID- 18429628 TI - Evaluation of cationic assemblies constructed with amino acid based lipids for plasmid DNA delivery. AB - We synthesized cationic lipids bearing lysine, histidine, or arginine as a cationic headgroup for use in gene transfer studies. The cationic assemblies formed from lysine- or arginine-type lipids gave unilamellar vesicles (approximately 100 nm diameter), whereas the morphology of the histidine-type lipids was tube-like. The competences of the cationic assemblies were sufficient to form lipoplexes, and the resulting lipoplexes were evaluated in terms of gene expression efficiencies with COS-7 cells. The lysine- or arginine-type lipids exhibited higher gene expression efficiencies than that of Lipofectamine2000, a conventional transgenic reagent, indicating that stable lipoplexes could be prepared between spherical cationic assemblies and plasmid DNA. The gene expression efficiency in relation to the cationic headgroup of the lipids was as follows: lysine > or = arginine > histidine. In addition, gene expression efficiency was enhanced by decreasing the length of the alkyl chain of the hydrophobic moiety. Unlike Lipofectamine2000, no reduction in transfection efficiency in the presence of fetal bovine serum was observed for the lipoplexes formed using synthetic cationic lipids. Moreover, the synthetic cationic lipids revealed remarkably low cytotoxicity compared with Lipofectamine2000. In conclusion, cationic assemblies formed from 1,5-ditetradecyl-N-lysyl-L-glutamate or 1,5-ditetradecyl-N-arginyl-L-glutamate can be used as an effective plasmid DNA delivery system. PMID- 18429629 TI - Self-assembling chimeric protein for the construction of biodegradable hydrogels capable of interaction with integrins expressed on neural stem/progenitor cells. AB - The poor survival of neural stem/progenitor cells following transplantation into the brain is the major problem limiting the effect of cell-based therapy for Parkinson's disease. To overcome this problem, we are involved in designing keratin-based hydrogels that serve as physical barriers to prevent the infiltration of inflammatory cells. Another feature of the hydrogels is to contain a polypeptide that promotes integrin-mediated cell adhesion. To construct such hydrogels, a chimeric protein consisting of an alpha-helical polypeptide and a globular domain derived from laminin was synthesized by means of recombinant DNA technology and coassembled with extracted keratins that form hydrogels through intermolecular coiled-coil association of alpha-helical segments. It was found that neurosphere-forming cells specifically adhered to the keratin-based composite hydrogel and actively proliferated at a high survival rate. These results suggested that the composite hydrogel provides microenvironments suitable for the survival and proliferation of neural progenitor cells. PMID- 18429630 TI - A novel polymer supported approach to nucleoside modification. AB - Polymer-supported O(6)-(benzotriazol-1-yl)inosine derivatives (Pol-I and Pol-dI) have been synthesized reasonably effectively via reaction of nucleoside phosphonium salts with polymer-linked HOBt (Pol-HOBt). In constast to solution chemistry, use of polymer-supported BOP (Pol-BOP) did not lead to efficient nucleoside loading. Presence of the nucleosides on the support could be readily detected by MAS NMR. Exposure of the polymer-supported nucleosides, Pol-I and Pol dI, to alcohol, phenol, thiol and amine nucleophiles caused cleavage from the support leading directly to the C-6 modified nucleoside analogues. To our knowledge, these are the first examples of the application of such technology for nucleoside modification. Where possible, results of reactions with the polymer supported nucleosides are compared to those from solution chemistry, providing insight into the differences between the two techniques. These new polymer supported nucleosides can be conveniently utilized for diversity-oriented synthesis. PMID- 18429631 TI - Novel pyromellitic diimide-based macrocycle with a linear pi-electronic system and bis(phenylethynyl)pyromellitic diimide: syntheses, structures, photophysical properties, and redox characteristics. AB - We report the syntheses, structures, photophysical properties, and redox characteristics of the [2 + 2] pyromellitic diimide-based macrocycle with a linear pi-electronic system 2 as well as the 3,6-bis(phenylethynyl)pyromellitic diimide derivative 3. The interesting solid state structural properties of the clathrates of 3 with pi-donors are also reported. The macrocycle 2 was synthesized by the direct cyclocondensation followed by the Sonogashira coupling reaction. X-ray crystallographic studies showed that the phenylacetylene moieties in 2 formed the intramolecular benzene dimer structures, and the bis(phenylethynyl)pyromellitic diimide moieties in both 2 and 3 were stacked in a parallel and slanted arrangement. Theoretical calculations for 2' and 3 suggested the existence of electrostatic interactions between the bis(phenylethynyl)pyromellitic diimide moieties. The UV/vis spectral measurements and TDDFT calculations of 2, 2', and/or 3 were performed to understand their electronic transitions. The fluorescence spectral measurements showed that 2 and 3 have visible fluorescence properties and 2 displays an excimer fluorescence at ca. 590 nm. The cyclic voltammetry measurements revealed that the electrostatic repulsion between the diimide moieties in 2 is greater than that in 1 according to the extension of the pi-electronic systems. X-ray crystallography of the clathrates of 3 with various pi-donors demonstrated the formation of the segregated donor-acceptor structures, indicating the strong aggregation ability of the bis(phenylethynyl)pyromellitic diimide moiety. PMID- 18429632 TI - Fluorogenic imines for fluorescent detection of Mannich-type reactions of phenols in water. AB - Fluorogenic imines and their precursor amines that can be used for fluorescent visualization of Mannich-type reactions of phenols in aqueous buffers have been developed. The precursor amines are aniline derivatives that are covalently conjugated to fluorophores. These amines and their imine derivatives were nonfluorescent or very weakly fluorescent. On the other hand, addition products of the imines to phenols showed more than 100-fold higher fluorescence than the imines and the precursor amines. PMID- 18429633 TI - CeCl3.7H2O: an effective additive in Ru-catalyzed enantioselective hydrogenation of aromatic alpha-ketoesters. AB - In the presence of catalytic amounts of CeCl 3.7H 2O, [RuCl(benzene)(S) SunPhos]Cl is a highly effective catalyst for the asymmetric hydrogenation of aromatic alpha-ketoesters. A variety of ethyl alpha-hydroxy-alpha-arylacetates have been prepared in up to 98.3% ee with a TON up to 10,000. Challenging aromatic alpha-ketoesters with ortho substituents are also hydrogenated with high enantioselectivities. The addition of CeCl 3.7H 2O not only improves the enantioselectivity but also enhances the stability of the catalyst. The ratio of CeCl 3.7H 2O to [RuCl(benzene)(S)-SunPhos]Cl plays an important role in the hydrogenation reaction with a large substrate/catalyst ratio. PMID- 18429634 TI - Application of 2-pyridyl-substituted hemithioindigo as a molecular switch in hydrogen-bonded porphyrins. AB - When the photochromism of 2-(3'-pyridylmethylene)-7-ethylbenzo[b]thiophen-3(2H) ones (4) was investigated, high thermal stability of the E isomer of 4, 4(E) and good repeatability of the photoinduced E,Z-isomerization were found. Association constants of the 1:1 complexations of 4(Z) and 4(E) with the ureidoporphyrin 1 and with the pentafluorobenzamidoporphyrin 2 were evaluated. We found that 1 captures 4(E) preferentially to 4(Z) and, reversely, 2 prefers 4(Z) to 4(E). On the basis of these differences in the binding ability, we concluded that the repeatable movement of the hemithioindigo, so-called the hemithioindigo shuttle, between two kinds of porphyrins was controlled by the photoirradiation. These movements were applied to create a molecular switch for changes in the quinone distribution between two kinds of porphyrins. PMID- 18429635 TI - Benzyl 2-cyano-3,3-dimethyl-1-pyrrolidinecarboxylate, a versatile intermediate for the synthesis of 3,3-dimethylproline derivatives. AB - The synthesis of racemic nitrile (+/-)-9 was accomplished in four steps and 58% overall yield from the known pyrrolidinone 5. Nitrile (+/-)-9 was resolved via preparative chiral HPLC to afford optically pure nitriles (+)-9 and (-)-9, from which 3,3-dimethylprolines (+)-1 and (-)-1 and 3,3-dimethylprolinamides (+)-2 and (-)-2 could be accessed in nearly quantitative yield, without loss of optical purity. The absolute configurations of the resolved prolines and prolinamides were determined by correlation with an intermediate of known absolute stereochemistry. PMID- 18429636 TI - Synthesis, structure, and E-Z isomerization of beta-(hetero)aryl-alpha-nitro alpha,beta-enals. AB - The first general methodology for the gram-scale preparation of previously overlooked beta-(hetero)aryl-alpha-nitro-alpha,beta-enals (3) is reported. Condensation of (hetero)aromatic aldehydes with 2-nitroethanol gave the E-isomers of uncommon beta-(hetero)aryl-alpha-hydroxymethyl-alpha,beta-unsatured nitroalkenes (2), as determined by NOE and X-ray studies. alpha-Nitro-alpha,beta enals 3 were subsequently obtained by hypervalent iodine oxidation of 2 as E-Z mixtures in solid form. They showed varied stability and solvent-dependent thermal-promoted and photopromoted E-Z interconversion. Starting with furfural, experimental conditions were developed to prepare the corresponding nitroenal 3a enriched in either the E or the Z isomer: E-3a/Z-3a approximately 90/10 and 20/80, respectively. In contrast with other structurally related compounds, nitroenals 3 have their (hetero)aryl-vinyl unit and their formyl and nitro groups all in a planar arrangement, both in solid form and in solution; accordingly, they are colored compounds with predicted high dipole moments. As deduced from solution-NMR and X-ray data, the C=C and the C=O double bonds in 3 are exclusively s- cis-oriented; this disposition corresponds in fact to the DFT computed most stable conformer. PMID- 18429638 TI - Increased hot-plate ignition probability for nanoparticle-laden diesel fuel. AB - The present study attempts to improve the ignition properties of diesel fuel by investigating the influence of adding aluminum and aluminum oxide nanoparticles to diesel. As part of this study, droplet ignition experiments were carried out atop a heated hot plate. Different types of fuel mixtures were used; both particle size (15 and 50 nm) as well as the volume fraction (0%, 0.1%, and 0.5%) of nanoparticles added to diesel were varied. For each type of fuel mixture, several droplets were dropped on the hot plate from a fixed height and under identical conditions, and the probability of ignition of that fuel was recorded based on the number of droplets that ignited. These experiments were repeated at several temperatures over the range of 688-768 degrees C. It was observed that the ignition probability for the fuel mixtures that contained nanoparticles was significantly higher than that of pure diesel. PMID- 18429639 TI - Electric field quenching of carbon nanotube photoluminescence. AB - The effect of external electric fields on the photoluminescence intensity of single-walled carbon nanotubes was investigated for individual nanotubes and bulk samples in polymeric films. Fields of up to 10(7) V/m caused dramatic, reversible decreases in emission intensity. Quenching efficiency varied as the cosine of the angle between the field and nanotube axis and decreased with increasing optical band gap. Photoluminescence intensity was found to follow a reciprocal hyperbolic cosine dependence on electric field. PMID- 18429640 TI - Enzymic thin film coatings for bioactive materials. AB - Bioactive materials have been explored for a broad range of applications including biocatalysts, biosensors, antifouling membranes and other functional and smart materials. We report herein a unique method for preparation of bioactive materials through a spin coating process. Specifically, we investigated the preparation of protease Subtilisin Carlsberg-coated plastic films and examined their activities for hydrolysis of chicken egg albumin (CEA). The process generated enzymic coatings with a typical loading of 13 microg/cm2, retaining 46% of the enzyme activity for hydrolysis of CEA in aqueous solutions. Interestingly, the surface-coated protease thin film not only catalyzed the hydrolysis of CEA in aqueous solutions, but also showed good activity for solid state CEA that was coated on top of the enzyme thin film. PMID- 18429641 TI - Study of detergent-mediated liberation of hepatitis B virus-like particles from S. cerevisiae homogenate: identifying a framework for the design of future generation lipoprotein vaccine processes. AB - Virus-like particles (VLPs) are expressed intracellularly in Saccharomyces cerevisiae and the recovery process involves the use of a detergent, which facilitates the release of VLP from host cell components. The detergent-mediated liberation of VLPs is a critical step in primary recovery and is responsible for setting the backdrop for subsequent purification in terms of product yield and characteristics of the process stream. In this paper the use of Triton X-100 detergent for the recovery of lipid envelope VLPs, using the hepatitis B surface antigen (HBsAg) as the VLP model, was investigated. To develop a framework that can be adopted in process design for future generation VLP vaccine candidates, the impact of Triton X-100 was characterized via different response factors: (i) recovery and activity of the HBsAg; (ii) level of protein and lipid contamination from the host cell; and (iii) indirect impact on the performance of an ultrafiltration step following primary recovery. Our studies identified that an increase in detergent concentration favors recovery of HBsAg only to a specific threshold, 0.5% v/v Triton X-100. Further increase in detergent results in delipidation of HBsAg leading to loss in antigenic activity. The level of contamination due to host protein and lipid co-liberation is in proportion with the amount of detergent employed. Greater membrane resistance during ultrafiltration was observed for samples generated using higher concentrations of detergent due to the increase in membrane fouling by the contaminants. Based on this study, Triton X-100 concentrations in the range of 0.2-0.5% v/v appears to be most suitable for recovery of native HBsAg. Choosing between 0.2-0.5% v/v would involve identifying a suitable tradeoff between desired product yield and the level of contamination that can be tolerated by downstream operations. PMID- 18429642 TI - Elephantiasis nostras verrucosa: a review. AB - Elephantiasis nostras verrucosa (ENV) is a rare and dramatic sequela of chronic nonfilarial lymphedema. The condition is characterized by papules, verrucous lesions, enlargement, and woody fibrosis of the affected area. ENV is a progressive condition, and, without intervention, ongoing deformity and disability will result. Management of ENV is often challenging, but a variety of successful medical and surgical treatment strategies have been reported. PMID- 18429637 TI - Enzymatic pathways that regulate endocannabinoid signaling in the nervous system. PMID- 18429643 TI - Pemphigus: a treatment update. AB - Pemphigus is an uncommon but potentially life-threatening chronic autoimmune bullous disorder. Antibodies are directed against antigens (desmoglein 1 and 3) in the desmosomes linking keratinocytes and against acetylcholine receptors. Conventional treatment with high-dose corticosteroids, sometimes with adjuvant immunosuppressive agents, may be associated with very serious adverse effects. There is an urgent need to establish the evidence for the safest and most effective form of treatment. A literature review has revealed 11 controlled (9 randomized) trials of treatment for pemphigus. The numbers of participants in the individual trials are small and the data cannot be pooled as they evaluate different forms of treatment. The results of these trials suggest that very high doses of corticosteroids, either as pulse therapy or in daily dosage, are not superior to moderate daily doses. Based on evidence from the available trials, addition of an immunosuppressive agent generally does not appear to offer substantial benefit in terms of clinical response. However, a recent study demonstrated a significant reduction in corticosteroid requirements among patients receiving immunosuppressive agents. Newer therapies, such as biologic agents (in particular rituximab), calcineurin inhibitors, or immunoadsorption appear promising but there are inadequate controlled trials to establish their role clearly. Initial open-label studies suggest that specific peptide immunotherapy may offer a safe and novel approach to the treatment of pemphigus in the future. At present, treatment of an individual patient with pemphigus requires clinical judgment and should not be based purely on guidelines or on the inadequate available evidence alone. There is an urgent need for large randomized, controlled, multicenter trials of treatment in patients with pemphigus. PMID- 18429644 TI - Chronic facial sarcoidosis including lupus pernio: clinical description and proposed scoring systems. AB - BACKGROUND: Facial lesions including lupus pernio are often a form of chronic cutaneous sarcoidosis. OBJECTIVE: To evaluate the intra- and inter-observer consistency of objective measures of chronic facial lesions. METHOD: This was a retrospective study of patients with chronic cutaneous facial lesions including lupus pernio. The lesions were evaluated using two methods. RESULTS: Of the 25 patients studied, 23 were women and 24 were African American. Lungs (24 patients), sinuses (11 patients), and eyes (7 patients) were also affected. The Sarcoidosis Activity and Severity Index (SASI) characterized individual areas of the face, with 95% of the observations being less than 2 points from the median. A facial SASI total gave a score for the entire face and 93.2% of the scores were within 3 points of the median. CONCLUSION: Patients with sarcoidosis and chronic facial lesions often have lung, sinus, and eye involvement. The SASI is a reproducible scoring system for chronic facial lesions. PMID- 18429645 TI - Occult herpes simplex virus colonization of bullous dermatitides. AB - BACKGROUND: Acantholytic disorders, including pemphigus vulgaris, chronic benign familial pemphigus (Hailey-Hailey disease, superficial pemphigus), Darier disease, and Grover transient acantholytic dermatosis, as well as other vesiculo bullous disorders, including bullous pemphigoid, epidermolysis bullosa, and atopic dermatitis, are prone to florid infections by herpes simplex virus (HSV)-I and -II, and, more rarely, by varicella-zoster virus (VZV). As these infections are difficult to recognize clinically and histologically, their frequency remains unknown. A possible occult viral colonization has never been documented in these disorders. The manner in which the primary bullous disorders are contaminated by herpesviridae remains unclear. OBJECTIVE: To retrospectively assess the possible presence of HSV and VZV in a series of biopsies of acantholytic disorders and bullous pemphigoid. METHOD: The typical alpha-herpesviridae-related cytopathic signs were searched for by conventional microscopy in skin biopsies of patients with bullous pemphigoid (n = 20), pemphigus vulgaris (n = 19), Darier disease (n = 18), chronic benign familial pemphigus (n = 3), and Grover transient acantholytic dermatosis (n = 3). Immunohistochemistry (IHC) targeted specific HSV I, HSV-II, and VZV antigens. Polymerase chain reaction (PCR) was used for detecting HSV- and VZV-specific DNA sequences. RESULTS: No cytopathic signs suggestive of HSV or VZV infection were detected. However, IHC revealed HSV antigens in Darier disease (1/18, HSV-I), Grover transient acantholytic dermatosis (1/3, HSV-I), pemphigus vulgaris (1/19, HSV-I), and bullous pemphigoid (2/20, HSV-I and HSV-II). In these IHC-positive cases, PCR amplified specific HSV primers in Darier disease (1/18), pemphigus vulgaris (1/19), and bullous pemphigoid (1/20). VZV antigens and nucleic acids were never identified. The HSV antigens were nearly always restricted to the upper part of the granular layer and thus differed from the usual HSV distribution during cutaneous infection. Negative and positive controls yielded consistently positive and negative results, respectively. CONCLUSION: This report shows for the first time that clinically and histologically occult HSV colonization may occur in Darier disease, Grover transient acantholytic disease, pemphigus vulgaris, and bullous pemphigoid. Given the frequent use of immunosuppressive treatments for primary bullous disorders, greater awareness of HSV colonization and infection is recommended in these patients. PMID- 18429646 TI - Safety and efficacy of low-dose bexarotene and PUVA in the treatment of patients with mycosis fungoides. AB - BACKGROUND: The new rexinoid bexarotene is a retinoid X receptor antagonist and immune response modifier. Although combinations of oral bexarotene and psoralen plus UVA (PUVA) have been tried in patients with all stages of mycosis fungoides (MF), the dosage of bexarotene used in these combination regimens has been variable. OBJECTIVE: To assess the efficacy and safety of low-dose oral bexarotene and PUVA in patients with relapsed or treatment-refractory MF following monotherapy with multiple agents including PUVA, narrow-band UVB, interferon-alpha, oral bexarotene, and topical corticosteroids. METHOD: Combination therapy with PUVA three times weekly and low-dose oral bexarotene (150 or 300 mg/day, depending on physicians' preference) was administered to 14 patients, seven men and seven women (median age 49.5 years, range 30-75 years), with relapsed or refractory MF stages I-III. All responders received maintenance treatment at the same bexarotene dose that induced remission until progression or unacceptable toxicity. RESULTS: Low-dose oral bexarotene combined with PUVA was associated with an overall response rate (complete response or partial response) in 67% of the nine patients with refractory MF who completed the treatment course. Of these nine patients, four had a complete response, two had a partial response, one had stable disease, and two had progressive disease. Five patients withdrew because of hyperlipidemia. Oral bexarotene was continued as maintenance therapy in three of the four complete responders (one refused); two of these patients relapsed 2-10 months after PUVA discontinuation. Patients with partial response or stable disease received the combination for 3-5 months and were switched to another treatment regimen because of lack of further response. Therapy was fairly well tolerated. CONCLUSION: In a select population of patients who had not responded to at least one monotherapy for early-stage MF, a combination of low-dose oral bexarotene and PUVA was successful in achieving a satisfactory overall response rate in 67% of patients who completed the treatment course and was fairly well tolerated. Limitations of the study include the small number of patients evaluated, its retrospective nature, and the fact that patients were commenced on different bexarotene starting doses (150 or 300 mg/day), depending on physicians' preference. PMID- 18429647 TI - Significant improvement in extensive lichen sclerosus with tacrolimus ointment and PUVA. AB - Lichen sclerosus is an uncommon, chronic inflammatory skin disorder of unknown origin. It is clinically characterized by sclerotic, whitish, atrophic-type lesions. The most frequent site of the lesions is generally the genital region, with about 15-20% having additional extragenital involvement. We present the case of a 62-year-old woman with a very extensive extragenital lichen sclerosus who showed great clinical and subjective improvement with 0.1% tacrolimus ointment and psoralen plus UVA (PUVA), without any topical or systemic adverse effects. Combined treatment of 0.1% tacrolimus ointment and PUVA may be a good option in extensive cases of lichen sclerosus or when other treatment options have failed, and has a good tolerability and safety profile. PMID- 18429648 TI - Metastasis of mediastinal epithelioid angiosarcoma to the finger. AB - Epithelioid angiosarcoma (EA) is a rare malignant, vascular tumor that is usually observed in middle-aged and elderly males. Cutaneous metastasis of EA is extremely rare. We report the case of a 41-year-old woman presenting with a painful bluish, bulla-like lesion on the distal extent of the left third finger. The patient had recently been diagnosed with mediastinal EA with disseminated metastases. The skin biopsy specimen revealed metastatic EA. This is thought to be the first reported metastasis of EA to the finger. Whenever a patient has metastatic disease, acrometastases should be considered in the differential diagnosis of inflammatory lesions of the digits and a skin biopsy should be performed. PMID- 18429649 TI - Superficial cutaneous leiomyosarcoma: a rare, misleading tumor. AB - Leiomyosarcomas are rare malignant tumors of smooth muscles. Superficial leiomyosarcoma is generally a disease of middle age, most frequently encountered between 40 and 60 years of age. It is usually diagnosed late or misdiagnosed, since it is a very rare tumor of the head and neck. Awareness of the particularly misleading features of this tumor, especially in elderly patients, is important, as delayed diagnosis is correlated with larger size and invasiveness into contiguous structures, which influence the practicability of radical resection. We present the case of an 81-year-old man with cutaneous leiomyosarcoma on the forehead. PMID- 18429650 TI - Acute developing generalized pustular rash. PMID- 18429651 TI - Clindamycin/benzoyl peroxide gel (BenzaClin): a review of its use in the management of acne. AB - Clindamycin 1%/benzoyl peroxide 5% (BenzaClin) is a combination gel indicated for use twice daily, or as directed by a physician, for the topical treatment of inflammatory and noninflammatory lesions of acne vulgaris. In well designed clinical trials in patients with mild to moderately severe acne, the efficacy of once- or twice-daily clindamycin/benzoyl peroxide in the reduction of inflammatory lesion counts was greater than that of benzoyl peroxide alone, clindamycin alone, or tretinoin plus clindamycin, and not significantly different from that of erythromycin/benzoyl peroxide. In the reduction of noninflammatory lesion counts, the efficacy of once- or twice-daily clindamycin/benzoyl peroxide was greater than that of clindamycin alone, but not significantly different to that observed with benzoyl peroxide, tretinoin plus clindamycin, or erythromycin/benzoyl peroxide. Clindamycin/benzoyl peroxide has a fairly rapid onset of action, with acne improvement usually recorded within 2-4 weeks. Despite widespread use, bacterial resistance is not associated with clindamycin/benzoyl peroxide. The product is generally well tolerated, and the main treatment-related adverse events in clinical trials were application-site dryness, irritation, peeling, and erythema. Thus, clindamycin/benzoyl peroxide is an effective and well tolerated option for the management of mild to moderately severe acne. PMID- 18429652 TI - Autochthonous creeping eruption in an Italian child. PMID- 18429653 TI - Cost-effectiveness analysis: developing nations left behind. PMID- 18429654 TI - Perspectives on patient-reported outcomes : content validity and qualitative research in a changing clinical trial environment. AB - Patient-reported outcomes (PROs) represent the voice of the patient in drug and device evaluation. As such, the outcomes selected for evaluation must be relevant to the patient, and the instruments used to capture them must have sound measurement properties. This paper discusses the role of qualitative research methods in assuring PRO clarity and content validity in a clinical trial environment characterized by evolving regulatory policy, new advances in technology and increasingly diverse samples within global clinical trials. Three important PRO measurement issues influenced by these changes are addressed: (i) endpoint selection and instrument development; (ii) the adaptation of instruments for electronic administration; and (iii) conceptual equivalence of measures across diverse samples. These are viewed as interrelated issues of content validity that can be addressed through the appropriate and rigorous application of qualitative research methods. Focus groups, cognitive debriefing, user acceptance testing and translation methodologies are discussed as ways to address and document the content validity of PRO instruments and ensure the empirical data representing the voice of the patient is sound. PMID- 18429655 TI - Economic evaluation of interventions to treat opiate dependence : a review of the evidence. AB - Opiate dependence imposes a significant economic burden on society in terms of treatment-related costs and prevention services, other healthcare costs, the work absenteeism of patients, productivity loss arising from premature death of patients, costs associated with crime, and social welfare expenditure. The objective of this research is to review the literature on economic evaluation of treatment of opiate dependence (including detoxification, maintenance and psychosocial support).A literature review was performed on several electronic databases, including MEDLINE (Ovid), Cochrane Database of Systematic Reviews, NHS Economic Evaluation Library Database (via Cochrane Library), Web of Science, Social Science Citations Index, EMBASE and PsycINFO. A sensitive approach was used in order to maximize the number of articles retrieved; no language or publication year limitations were applied to the searches. A combination of subject heading term searches and natural word searches were used. The Drummond checklist was applied to assess the quality of economic evaluations.A total of 259 articles were considered relevant, with eight review studies identified. The treatment spectrum ranged from detoxification to maintenance treatments involving the use of agonist and/or antagonist treatments. The evidence suggests that, although the quality of economic evaluations is reasonably good, there is a dearth of knowledge about the cost effectiveness of treatments for opiate dependence. The majority of the literature reporting the results of cost effectiveness analyses used surrogate outcome measures and adopted a narrow treatment provider perspective. Studies that have conducted cost-benefit analyses, in spite of methodological divergences, generally adopted a societal perspective and consistently demonstrated positive economic returns from opiate treatment. A paucity of research examined the extent to which psychosocial or behavioural interventions support or replace conventional pharmacological approaches. Economic evaluation provides a useful framework to assist policy makers in allocating resources across competing needs. Opiate dependence is a considerable burden on society's resources, and treatment provides a cost beneficial solution to address these consequences. However, to better inform the decision-making process, researchers must continue to produce high-quality, methodological, comparable and scientifically credible economic evaluations. PMID- 18429656 TI - Economic evaluations in rheumatoid arthritis: a critical review of measures used to define health States. AB - We reviewed the clinical measures used in rheumatoid arthritis (RA) economic evaluations with respect to their relevance and sensitivity to changes in survival, health-related quality of life (HR-QOL) and costs. We compared the measures from the economic perspective and discussed the validity of methods used to extrapolate beyond the trial data. Cost-effectiveness evaluations of disease modifying antirheumatic drugs in RA were identified by searching MEDLINE, EMBASE, Econlit and NHS EED databases. Studies were retained if they extrapolated beyond randomized controlled trial evidence using relationships between clinical measures, costs and utilities. In the 22 studies identified, clinical severity was measured using the Health Assessment Questionnaire (HAQ) Disability Index, the American College of Rheumatology (ACR) response criteria, the Disease Activity Score (DAS) or a combination of the HAQ and DAS. The HAQ is correlated with mortality, costs and HR-QOL instruments, and several studies used linear relationships to model these associations. However, a polynomial relationship or discrete states may be more appropriate for patients at the extremes of the disease spectrum, and numerous HAQ health states may be required to capture differences in mortality risk. While the ACR response criteria is a more comprehensive measure than the HAQ, it is a relative measure, which creates difficulties when estimating absolute changes in HR-QOL, costs and mortality risk. The evidence base linking DAS scores with HR-QOL instruments, costs and mortality is less robust, possibly due to the comparatively recent development of the measure and the limited number of possible scores (mild/moderate/severe). While there is some evidence of a relationship between DAS scores and costs, the DAS does not capture all aspects of HR-QOL, and no significant relationship has been established with mortality risk. Evidence suggests the HAQ to be the primary clinical measure for use in economic evaluations as it is measured in almost all clinical studies, and is closely correlated to health utilities, mortality and costs. While new developments suggest the sensitivity of health states may be improved by combining the HAQ with measures such as the DAS, further research is required in this area. Further research is also required to explore the advantages in using either continuous or discrete health states. PMID- 18429657 TI - Methodological issues in evaluating cost effectiveness of adjuvant aromatase inhibitors in early breast cancer: a need for improved modelling to aid decision making. AB - The optimal adjuvant hormonal strategy in post-menopausal women with early breast cancer is a subject of ongoing debate. Aromatase inhibitors (AIs) have been successfully evaluated in clinical trials that have compared them with a standard treatment of 5 years of tamoxifen. However, several options are available in terms of treatment schedule and selected drug. Systematic reviews of clinical trials and health economic evaluations attempt to contribute to the debate. The objective of this paper is to provide a critical review of existing health economic evaluations with a focus on those parameters and assumptions with the largest impact on final outcomes.A wide range of different inputs and assumptions exist, which make a comparison of results difficult, if not impossible. In particular, the modelling of recurrence rates over longer time horizons than those observed in clinical trials, a cornerstone of health economic modelling, is subject to quite different approaches. The practice of indirect comparison of different AIs without sufficiently acknowledging population differences is also bothersome. A list of key features (related to time horizon, clinical data input, patient subtypes, budget impact and model calibration) that an ideal model should have in order to better assist decision makers in this field is proposed. PMID- 18429658 TI - A multicentre, prospective study to evaluate costs of septic patients in Brazilian intensive care units. AB - BACKGROUND: Sepsis has a high prevalence within intensive care units, with elevated rates of morbidity and mortality, and high costs. Data on sepsis costs are scarce in the literature, and in developing countries such as Brazil these data are largely unavailable. OBJECTIVES: To assess the standard direct costs of sepsis management in Brazilian intensive care units (ICUs) and to disclose factors that could affect those costs. METHODS: This multicentre observational cohort study was conducted in adult septic patients admitted to 21 mixed ICUs of private and public hospitals in Brazil from 1 October 2003 to 30 March 2004. Complete data for all patients admitted to the ICUs were obtained until their discharge or death. We collected only direct healthcare-related costs, defined as all costs related to the ICU stay. Enrolled patients were assessed daily in terms of cost-related expenditures such as hospital fees, operating room fees, gas therapy, physiotherapy, blood components transfusion, medications, renal replacement therapy, laboratory analysis and imaging. Standard unit costs (year 2006 values) were based on the Brazilian Medical Association (AMB) price index for medical procedures and the BRASINDICE price index for medications, solutions and hospital consumables. Medical resource utilization was also assessed daily using the Therapeutic Intervention Scoring System (TISS-28). Indirect costs were not included. RESULTS: With a mean (standard deviation [SD]) age of 61.1 +/- 19.2 years, 524 septic patients from 21 centres were included in this study. The overall hospital mortality rate was 43.8%, the mean Acute Physiology And Chronic Health Evaluation II (APACHE II) score was 22.3 +/- 5.4, and the mean Sequential Organ Failure Assessment (SOFA) score at ICU admission was 7.5 +/- 3.9. The median total cost of sepsis was $US 9632 (interquartile range [IQR] 4583-18 387; 95% CI 8657, 10 672) per patient, while the median daily ICU cost per patient was $US 934 (IQR 735-1170; 95% CI 897, 963). The median daily ICU cost per patient was significantly higher in non-survivors than in survivors, i.e. $US 1094 (IQR 888-1341; 95% CI 1058, 1157) and $US 826 (IQR 668-982; 95% CI 786, 854), respectively (p < 0.001). For patients admitted to public and private hospitals, we found a median SOFA score at ICU admission of 7.5 and 7.1, respectively (p = 0.02), and the mortality rate was 49.1% and 36.7%, respectively (p = 0.006). Patients admitted to public and private hospitals had a similar length of stay of 10 (IQR 5-19) days versus 9 (IQR 4-16) days (p = 0.091), and the median total direct costs for public ($US 9773; IQR 4643-19 221; 95% CI 8503, 10 818) versus private ($US 9490; IQR 4305-17 034; 95% CI 7610, 11 292) hospitals did not differ significantly (p = 0.37). CONCLUSIONS: The present study provides the first economic analysis of direct costs of sepsis in Brazilian ICUs and reveals that the cost of sepsis treatment is high. Despite similar ICU management, there was a significant difference regarding patient outcome between private and public hospitals. Finally, the median daily costs of non-survivor patients were higher than survivors during ICU stay. PMID- 18429660 TI - Energy metabolism of sea urchin spermatozoa: an approach based on echinoid phylogeny. PMID- 18429659 TI - Updated estimates of survival and cost effectiveness for imatinib versus interferon-alpha plus low-dose cytarabine for newly diagnosed chronic-phase chronic myeloid leukaemia. AB - BACKGROUND: For trials in which participants are followed beyond the main study period to assess long-term outcomes, economic evaluations conducted using short term data should be systematically updated to reflect new information. METHODS: We used 60-month survival data from the IRIS (International Randomized study of Interferon vs STI571) trial to update previously published cost-effectiveness estimates, based on 19 months of follow-up, of imatinib versus interferon (IFN) alpha plus low-dose cytarabine in patients with chronic-phase chronic myeloid leukaemia. For patients treated with imatinib, we used the 60-month data to calibrate the survival curves generated from the original cost-effectiveness model. We used historical data to model survival for patients randomized to IFNalpha. We updated costs for medical resources using 2006 Medicare reimbursement rates and applied average wholesale prices (AWPs) and wholesale acquisition costs (WACs) to study medications. RESULTS: Five-year survival for patients randomized to imatinib was better than predicted in the original model (89.4% vs 83.2%). We estimated remaining life expectancy with first-line imatinib to be 19.1 life-years (3.8 life-years over the original model) and 15.2 QALYs (3.1 QALYs over the original estimate). Estimates for IFNalpha remained at 9.1 life-years and 6.3 QALYs. When we applied AWPs to study medications, incremental cost-effectiveness ratios (ICERs) were $US 51,800-57,500 per QALY. When we applied WACs, ICERs were $US 42,000-46,200 per QALY. CONCLUSION: Although the analysis revealed that the original survival estimates were conservative, the updated cost-effectiveness ratios were consistent with, or slightly higher than, the original estimates, depending on the method for assigning costs to study medications. PMID- 18429661 TI - Ionic dependence of the axotomy-induced long-lasting firing in an identified crayfish motoneuron. AB - I have reported previously that axotomy of an identifiable anal motoneuron of crayfish Procambarus clarkii induces a long-lasting firing and that a prolonged depolarizing pulse to its cut end can induce a similar response. In this study, I confirmed that this stimulus is comparable to axotomy; the frequency of stimulus induced firing increases linearly with the stimulus intensity and its firing pattern is the same as that following axotomy. Then, when the cut end was bathed for more than 1 hr in test solutions, it was examined whether the stimulus to the cut end induces or blocks the response. Na(+)-free saline (Tris(+) replaced Na(+)) or TTX (3 x 10(-7) M) reversibly blocked the response within 30 min. By contrast, Mn(2+) saline (40 mM Mn(2+) replaced Ca(2+)) or Ca(2+)-free salines (Mg(2+) or 1 mM EDTA replaced Ca(2+)) cannot block the response, but instead increased the firing frequency. These results obtained with stimulus were confirmed also by those with axotomy. I concluded that axotomy-induced firing, which occurs locally at its cut region, is primarily responsible for voltage dependent Na(+) conductances, but not for Ca(2+) ones. PMID- 18429662 TI - Production of the oocyte maturation-inducing substance of starfish by heat treatment of s-adenosylmethionine. AB - 1-Methyladenine (1-MeA) has been identified as the oocyte maturation-inducing substance (MIS) in starfish, but little is known about its biosynthesis. This study showed that starfish MIS activity was present in a reactant derived from S adenosylmethionine (SAM) by heat treatment. In vitro MIS production was markedly dependent on the temperature of the SAM solution: it increased as the temperature was raised, and reached a plateau within 5 min upon boiling, although hardly only MIS was observed upon incubation below 20 degrees C. MIS production was also dependent on the solution pH. Analyses by high-performance liquid chromatography and thin-layer chromatography showed that the MIS was 1-MeA, though the maximum amount of 1-MeA obtained from SAM by boiling was only 0.3% of the initial SAM amount. Furthermore, use of S-[(14)C-methyl]SAM showed that a methyl group of 1 MeA was transferred from the SAM. Thus, it is possible that 1-MeA may be produced from SAM in vivo. PMID- 18429663 TI - Sex-Linked Inheritance of the lf Locus in the Medaka Fish (Oryzias latipes). AB - The lf (leucophore free) locus was previously reported autosomal recessive in the medaka fish (Oryzias latipes). However, extensive linkage analyses in this study using various strains revealed that the lf locus was closely sex-linked. The recombination frequency between lf and the male determining factor (y) was 2.2% (10 recombinants out of 464 progeny). Because the lf/lf homozygous fish do not have visible leucophores, they are distinguishable from wild type in early developmental stages. In the Qurt strain with heterozygous sex chromosomes (X(lf)/X(lf) in females and X(lf)/Y(+) in males), we can predict sex of each embryo on second day after fertilization. The strain should be a very useful material for studying sex determination or differentiation mechanisms in the medaka fish. PMID- 18429664 TI - Effects of the Consumption of Male Spermatophylax on the Oviposition Schedule of Females in the Decorated Cricket, Gryllodes sigillatus. AB - The effects of the consumption of the spermatophylax produced by males on female fitness were studied in the decorated cricket, Gryllodes sigillatus. An increase in the number of spermatophylaces presented to females did not increase the total number of eggs made by females, the number of eggs laid, or the hatchability of eggs laid by females, but increased the number of eggs laid in the early stage of adult life of females. The duration of the egg stage decreased with the number of spermatophylaces presented to females. The implication of the results on the sham hypothesis that the spermatophylax does not have nutritional value is discussed. PMID- 18429665 TI - The Behavior of Heterolepidoderma sp. (Gastrotricha). AB - The behavior of Heterolepidoderma sp. was studied with the same approach as those already used for many species of ciliates. The ethogram we drew comprehends both helicoidal swimming (n = 20, r = 52.5 +/-12.2 mum, pitch = 512 +/- 101 mum, v--> = 215 +/- 43 mum/sec), periodically interrupted by irregular patterns changing the direction of the swimming of random angles and creeping on the substrate. The latter behavioral state, very common for the species we studied, occurs along tracks formed by successive elements (circular, C, vs linear segments, S) joined to each other by two kinds of reactions, which change their trajectory. The surprising similarities and the unexpected differences between the behavior of this gastrotrich and those of the ciliates already studied from this point of view are discussed, on the basis of the dimensional ranges and ecological niches shared by these two, definitely unrelated groups of organisms. PMID- 18429666 TI - A new species of corynoppia (acari, oribatida, oppiidae) from biscay (the basque country, northern Spain). AB - A new oribatid mite belonging to the family Oppiidae is described from the province of Biscay in the Basque Country, Northern Spain. We propose the name Corynoppia papillisetigera for it, whose shape and size of setae ta (c2) bearing papillae is the main and distinctive feature to separate it from allied species, such as Corynoppia foliatoides . For the moment it has been found in a coastal meadow and eucaliptus, evergreen-oak, and pine forests. PMID- 18429667 TI - Comparative studies of the genus echinometra from okinawa and mauritius. AB - Okinawan sea urchins, the genus Echinometra, are four independent species. But which species are the same species as E. mathaei and E. oblonga as described by is still open to question. To answer this question, a field survey of genus Echinometra was made in Mauritius (the type locality of E. mathaei) according to the characteristics used to classify Okinawan Echinometra: appearance, pore pairs, spicules in gonads and tubefeet, sperm shape, and distribution on a reef. The results of crossfertilization between Echinometra from Mauritius and Okinawa are also reported. Mauritian Echinometra are classified into three groups which resemble Okinawan Echinometra sp. B, D, and violet spine color Echinometra. The latter has almost the same characteristics as Okinawan Echinometra sp. B but with violet spines, a spine color not found in Okinawa. In cross-fertilization experiments, the sperm of Mauritian Echinometra sp. B-like and violet Echinometra fertilized Okinawan Echinometra sp. B with almost 100% success. However, fertilization was unsuccessful with other Okinawan Echinometra species. Therefore, it could be said that Okinawan Echinometra sp. B is the same as Mauritian Echinometra sp. B-like, and the counterparts of Okinawan Echinometra sp. A and C are not distributed in Mauritius. The descriptions of E. mathaei most match Mauritian Echinometra sp. B-like and the type locality of E. mathaei is Mauritius. Thus, it is probable that Mauritian Echinometra sp. B-like is E. mathaei. Therefore, Okinawan Echinometra sp. B, which most resembles Mauritian Echinometra sp. B-like, would be also E. mathaei. Echinometra sp. A and C would be considered to be new species. Echinometra sp. D is thought to be the same species as Mauritian black Echinometra. Whether Okinawan Echinometra sp. D is the same as E. oblonga remains as a problem for future research. Because the type locality of E. oblonga is not known and, it is suggested that the E. oblonga described in the Indo-West Pacific is a complex species. PMID- 18429668 TI - Cardioacceleratory Neurons of the Isopod Crustacean, Ligia exotica: Visualization of Peripheral Projection onto the Heart Muscle. AB - Innervation of the heart muscle by the cardioacceleratory neurons was morphologically and electrophysiologically examined in the isopod crustacean, Ligia exotica. Intracellular injection of neurobiotin into the first and second cardioacceleratory neurons (CA1 and CA2) revealed their peripheral axonal projections. Inside the heart, the CA1 and CA2 axons ran along the trunk of the cardiac ganglion. Finely arborized branches with many varicosities arose from the axon and projected over the heart muscle. Stimulation of either the CA1 or CA2 axon caused an overall depolarization in the muscle of a quiescent heart. The amplitude of the depolarization increased with increasing stimulus frequency. During stimulation, the membrane resistance of the heart muscle decreased. In a beating heart, the cardioacceleratory nerve stimulation caused multiple effects on the heart muscle activity and the heartbeat. The results suggest that the cardioacceleratory neurons of Ligia exotica regulate the amplitude of the heartbeat (inotropic effect) and the heart tonus (tonotropic effect) via the synaptic contacts on the heart muscle, while the heartbeat frequency (chronotropic effect) is regulated via the synapses on the cardiac ganglion neurons. PMID- 18429669 TI - Evidences for direct involvement of microtubules in cleavage furrow formation in newt eggs. AB - This paper aims at examining the effect of colchicine, a microtubular poison, on the process of furrow formation in whole eggs and egg fragments as well as the process of artificial induction of furrow-like dents, in eggs of the newt, Cynops pyrrhogaster. To apply colchicine locally to eggs, the eggs were slit across or along a furrow in a colchicine solution during first cleavage. When a slit was made across or in front of a growing furrow at the onset of its growth, the furrow quickly ceased growing and often regressed. Cortices containing an entire growing furrow were isolated along with a thin layer of subcortical cytoplasm immediately after the start of the first cleavage. Furrows in the cortices degenerated when the cortices were cultured in a colchicine solution, whereas they continued growing when they were cultured in Holtfreter's saline. Furrow inducing cytoplasm was injected to a site beneath the cortex in the animal half of the egg during first cleavage. When a small slit was made close to the site of the injection in a colchicine solution, no furrow-like dent was induced. These results imply that microtubules are directly involved in the generation and growth of cleavage furrows. PMID- 18429670 TI - The Spawning and Early Development of the Hawaiian Acorn Worm (Hemichordate), Ptychodera flava. AB - The spawning and early embryogenesis of the hemichordate, Ptychodera flava, in Hawaii are described in detail and illustrated with photographs of living material. Natural spawning in the evenings of early December was induced by a shift of seawater temperature from about 22 degrees C to about 26 degrees C. The fertilized egg divides equally and slowly at first, reaching 8 cells at about 5 hr after insemination at room temperature (20-24 degrees C). Divisions then appear to become slightly unequal and by 9 hr the embryo has divided into about 100 cells. The blastocoel forms during cleavage as an irregular space that, when viewed from the side, tends to appear oblate and ultimately appear crescent shaped as the vegetal plate thickens into the blastocoel. The archenteron forms at about 18 hr as a cleft beginning at the vegetal pole and extending into the vegetal plate. As development proceeds, the embryo expands and by 24 hr forms a typical deuterostome gastrula with an outer sphere of ectoderm and a inner tube of endoderm connected at the blastopore. An out-pocketing of the gut appears at the tip of the archenteron over the next 4 hr to form the protocoel which will become the proboscis coelom. Approaching 40 hr the gut becomes asymmetric and over the next few hr contacts the ectoderm to form a mouth. Hatching occurs during this time at about 45 hr of development. Morphogenesis continues to produce an early tornaria larva by about 60 hr. PMID- 18429671 TI - Multilaminar/ Vesicular Bodies Accumulating Glycoconjugates in Primary Spermatocytes of Cricket, Gryllus bimaculatus. AB - Lectin cytochemistry was carried out on thin sections of 6th-instar cricket testis using two GalNAc-specific lectins, Dolichos biflorus agglutinin (DBA) and Helix pomatia agglutinin (HPA), and the binding sites in primary spermatocytes were surveyed. Gold particles showing DBA-binding are observed specifically in dense-body clusters. These bodies are about 100-300 nm in diameter and exhibit multilaminar or multivesicular structure. HPA can bind to the dense-body clusters and another kind of larger multivesicular structures. These bodies seem to contain some heterogeneous substances, and sometimes show an autophagosome-like structure. The ultrastructures of these organelles surveyed in conventional Epon sections confirmed the structures of these multilaminar/vesicular bodies in cricket spermatocytes, which may play certain roles in intracellular circulation and degradation of glycoconjugates. PMID- 18429672 TI - Density-dependent regulation of the sex ratio in an annual plant. AB - Sex ratios are subject to strong frequency-dependent selection regulated by the mating system and the relative male versus female investment. In androdioecious plant populations, where males co-occur with hermaphrodites, the sex ratio depends on the rate of self-fertilization by hermaphrodites and on the relative pollen production of males versus hermaphrodites. Here, we report evolutionary changes in the sex ratio from experimental mating arrays of the androdioecious plant Mercurialis annua. We found that the progeny sex ratio depended strongly on density, with fewer males in the progeny of plants grown under low density. This occurred in part because of a plastic adjustment in pollen production by hermaphrodites, which produced more pollen when grown at low density than at high density. Our results provide support for the prediction that environmental conditions govern sex ratios through their effects on the relative fertility of unisexual versus hermaphrodite individuals. PMID- 18429673 TI - The cancer stem cell-vascular niche complex in brain tumor formation. AB - The cancer stem cell (CSC) theory hypothesizes that a small subpopulation of cells within a tumor mass is responsible for the initiation and maintenance of the tumor. The idea that brain tumors arise from this specific subset of self renewing, multipotent cells that serve as the locus for tumor formation, has gained great support as evidenced by recent advancements in the biology of breast and colon cancer. It is well established that recruitment of bone marrow-derived proangiogenic progenitor cells and angiogenesis are key events in the process of brain tumor formation; however, the orchestration of these events by the CSC population has only recently been unveiled. In this review, we first introduce the CSC theory and examine the functional development of the vascular niche, its purpose, constituents, and contribution to the development of the CSC-vascular niche complex. Through this discussion, we aim to shed light on the events that may be targeted for therapeutic intervention. PMID- 18429674 TI - Different ranking of avian colors predicted by modeling of retinal function in humans and birds. AB - Abstract: Only during the past decade have vision-system-neutral methods become common practice in studies of animal color signals. Consequently, much of the current knowledge on sexual selection is based directly or indirectly on human vision, which may or may not emphasize spectral information in a signal differently from the intended receiver. In an attempt to quantify this discrepancy, we used retinal models to test whether human and bird vision rank plumage colors similarly. Of 67 species, human and bird models disagreed in 26 as to which pair of patches in the plumage provides the strongest color contrast or which male in a random pair is the more colorful. These results were only partly attributable to human UV blindness. Despite confirming a strong correlation between avian and human color discrimination, we conclude that a significant proportion of the information in avian visual signals may be lost in translation. PMID- 18429676 TI - Health care-associated pneumonia (HCAP): a critical appraisal to improve identification, management, and outcomes--proceedings of the HCAP Summit. AB - Increasingly, patients are receiving treatment at facilities other than hospitals, including long-term-health care facilities, assisted-living environments, rehabilitation facilities, and dialysis centers. As with hospital environments, nonhospital settings present their own unique risks of pneumonia. Traditionally, pneumonia in these facilities has been categorized as community acquired pneumonia (CAP). However, the new designation for pneumonias acquired in these settings is health care-associated pneumonia (HCAP), which covers pneumonias acquired in health care environments outside of the traditional hospital setting and excludes hospital-acquired pneumonia (HAP), ventilator associated pneumonia (VAP), and CAP. Although HCAP is currently treated with the same protocols as CAP, recent evidence indicates that HCAP differs from CAP with respect to pathogens and prognosis and, in fact, more closely resembles HAP and VAP. The HCAP Summit convened national infectious disease opinion leaders for the purpose of analyzing current literature, clinical trial data, diagnostic considerations, therapeutic options, and treatment guidelines related to HCAP. After an in-depth analysis of these areas, the infectious disease investigators participating in the summit were surveyed with regard to 10 clinical practice statements. The results were then compared with results of the same survey as completed by 744 Infectious Diseases Society of America members. The similarities and differences between those survey results are the basis of this publication. PMID- 18429680 TI - Global spread and persistence of dengue. AB - Dengue is a spectrum of disease caused by four serotypes of the most prevalent arthropod-borne virus affecting humans today, and its incidence has increased dramatically in the past 50 years. Due in part to population growth and uncontrolled urbanization in tropical and subtropical countries, breeding sites for the mosquitoes that transmit dengue virus have proliferated, and successful vector control has proven problematic. Dengue viruses have evolved rapidly as they have spread worldwide, and genotypes associated with increased virulence have expanded from South and Southeast Asia into the Pacific and the Americas. This review explores the human, mosquito, and viral factors that contribute to the global spread and persistence of dengue, as well as the interaction between the three spheres, in the context of ecological and climate changes. What is known, as well as gaps in knowledge, is emphasized in light of future prospects for control and prevention of this pandemic disease. PMID- 18429681 TI - Balloon dilatation versus Amplatz dilatation for nephrostomy tract dilatation. AB - PURPOSE: In this study, we present our experience using balloon and Amplatz dilatation to establish a percutaneous tract. We also discuss advantages and risk factors of both techniques. MATERIALS AND METHODS: We retrospectively reviewed medical records of 229 patients who had undergone 235 percutaneous nephrolithotomy procedures. The nephrostomy tract had been dilated using a balloon (42 patients) or Amplatz (187 patients) dilator. Total operating time, preoperative and postoperative hemoglobin concentrations, number of tracts required, stone burden, blood transfusion rates, tract dilatation failures, and the cost of the dilatation system were compared between the groups. RESULTS: There were no statistically significant differences in operative time (85.7+/ 43.2 v 86.3+/-41.2 minutes; P=0.42), preoperative hemoglobin concentration (14.1+/-1.1 v 13.8+/-1.4 mg/dL; P= .153), postoperative hemoglobin concentration (11.6+/-1.7 v 11.2+/-1.5 mg/dL; P= .601), or blood transfusion rate (18.6% v 21.3%; P= .687) between the two groups. Also, there were no differences in failure rates between the two groups. CONCLUSIONS: The Amplatz dilator is comparable with the balloon dilator with regard to efficacy, speed, and safety. The Amplatz dilator is more cost-effective than the balloon dilator. However, kidney hypermobility may be a significant problem during Amplatz dilatation. PMID- 18429682 TI - Treatment of large impacted proximal ureteral stones: randomized comparison of percutaneous antegrade ureterolithotripsy versus retrograde ureterolithotripsy. AB - PURPOSE: We compared the safety and efficacy of percutaneous antegrade ureterolithotripsy with retrograde ureterolithotripsy for large impacted proximal ureter stones in a prospective randomized manner. MATERIALS AND METHODS: A total of 91 patients with large impacted proximal ureteral stones, defined as stones >1 cm in size located between the ureteropelvic junction and the lower border of the fourth lumbar vertebra, were prospectively randomized for antegrade (44) or retrograde (47) ureterolithotripsy. Failure of the procedure (conversion to an open procedure), intraoperative and postoperative morbidity, operative time, hospital stay, stone clearance at discharge home, and follow-up were analyzed in each group. RESULTS: The main complications were bleeding (2.3%; 1 of 43) for the antegrade procedure and ureteral injury (2.3%; 1 of 44) for the retrograde procedure. Percutaneous antegrade ureterolithotripsy was associated with longer operative times (75.4+/-11.8 v 30.6+/-7.8 minutes; P<0.001), longer hospital stay (6.3+/-0.5 v 2.1+/-0.4 days; P<0.001), and a longer interval to return to normal activities (7.8+/-0.7 v 2.7+/-0.6 days; P<0.001). Nevertheless, the percutaneous antegrade procedure had a higher stone-free rate both at discharge home (95.3% v 79.5%; P=0.027), and 1 month post-procedure (100% v 86.4%; P=0.026). CONCLUSIONS: Percutaneous antegrade ureterolithotripsy is a valuable treatment modality for impacted proximal ureteral calculi larger than 1 cm, and achieves higher stone free rates than those of retrograde ureteroscopy with holmium:YAG laser lithotripsy. The drawbacks of the antegrade procedure are longer operative time and hospital stay. PMID- 18429683 TI - Hemostatic properties of four devices for partial nephrectomy: a comparative ex vivo study. AB - PURPOSE: Laparoscopic partial nephrectomy (LPN) is a common minimally-invasive treatment modality for renal tumors, and achieving hemostasis during excision is a major challenge. The aim of our study was to investigate the hemostatic potential of four different devices for realizing this under standardized conditions. MATERIALS AND METHODS: LPN was performed on a standardized model of blood-perfused ex-vivo porcine kidneys. Each of the four devices (Greenlight KTP laser, Habib Sealer, LigaSure, and SonoSurg) as well as a scalpel (for comparison) were used to perform 10 excisions with the renal artery and vein clamped, and another 10 were performed with no clamping. Treatment time (TT), blood loss (BL), and the ease of handling of the device were measured and histologic examination of the margins was carried out. RESULTS: In general, TT was faster and there was less BL with clamping than without in all cases. TT was shortest for the KTP laser (6.07+/-1.2 minutes; P<0.0001), followed by the LigaSure (8.78+/-0.42 minutes), the SonoSurg (15.9+/-1.28 minutes), and the Habib (21.7+/-3.4 min). The SonoSurg showed a significantly higher BL without clamping (66+/-6 ml, p<0.0001) but there were no significant differences between the other devices. With clamping, BL four all four devices was comparable (13+/-2 ml) and without statistical significance (p=0.5). TT was shortest for the KTP laser (3.27+/-0.55 min, p<0.0001) followed by the LigaSure (6.47+/-0.38 s), the SonoSurg (8.35+/-3 min) and the Habib (9.71+/-1.18 minutes). The excised surface was completely coagulated for all of the devices except for the SonoSurg. CONCLUSION: Our ex-vivo study suggests that hemostatic potential and the coagulative effect of all four devices is inadequate. Furthermore, none of the devices produced clean and sharp resection margins, which is a prerequisite for negative surgical margins. PMID- 18429684 TI - Retrieval of migrated ureteral stents by coaxial cannulation with a flexible ureteroscope and paired helical basket. AB - PURPOSE: Retrieval of a proximally migrated ureteral stent is a technically challenging endoscopic procedure. We describe the use of a paired wire helical stone retrieval basket through a flexible ureteroscope to remove proximally migrated ureteral stents. MATERIALS AND METHODS: Five ureteral stents were lost in the proximal (1) and distal (4) ureter after lithotripsy or stent exchange. Once the location of the distal aspect of the ureteral stent was identified by either fluoroscopy or ureteroscopy, a safety guidewire was placed alongside the migrated stent. A flexible ureteroscope was brought to the distal end of the stent under direct visualization. Through the working port of the ureteroscope, the paired wire helical basket was deployed to grasp or coaxially cannulate the lumen of the migrated stent. Once inside the stent, the basket was then opened. The outward resistance of the deployed basket is sufficient to provide enough lateral force to reposition the stent into the bladder. RESULTS: All stents were successfully retrieved without complication. No patient had visual evidence of ureteral injury, and all patients were discharged on the same day as their procedure. CONCLUSION: While proximal migration of a ureteral stent is usually caused by technical error, the ideal technique to reposition the stent should require the least time, trauma to the ureter, and expense to the patient. We describe a simple technique to retrieve migrated stents anywhere along the urinary tract. PMID- 18429685 TI - Laparoscopic heminephrectomy in L-shaped crossed fused ectopia. AB - L-shaped fused ectopic kidney is a rare variant of crossed fused renal ectopia. We describe a case of crossed fused L-shaped kidney with congenital ureteropelvic junction obstruction involving an orthotopically located kidney presenting as renal trauma, and its management by laparoscopic heminephrectomy. PMID- 18429686 TI - Laparoscopic visualization and dissection of retroperitoneal lymph nodes after patent blue dye injection: a pilot study. AB - PURPOSE: Retroperitoneal lymph node dissection (RPLND) for testicular cancer is an important treatment modality for patients with stage I or IIA disease. Several urologists have previously reported the feasibility and usefulness of laparoscopic RPLND for such patients. The aim of this experimental pilot feasibility study was to investigate whether visualization of retroperitoneal lymph nodes with patent blue violet (PBV) dye application is a feasible and an effective method during laparoscopic RPLND in a pig model. MATERIALS AND METHODS: Four 12-month-old white male pigs were included in the study. After PBV dye injection into the spermatic funicular and intratesticular parenchyma, the color changes in the retroperitoneal region were examined during transperitoneal laparoscopic visualization of the retroperitoneum. The time interval between the injection and the staining of lymphatic structures was measured for each intervention. Blue-stained retroperitoneal nodal tissues were dissected and removed by the laparoscopic approach and histologic examination was performed. RESULTS: After PBV dye injection, intense staining of the ipsilateral retroperitoneal lymphatic vessels and nodes was seen. Distribution of the PBV and the color changes of the retroperitoneal lymphatic structures were examined under laparoscopic vision in all pigs. All blue-stained retroperitoneal nodular tissues were removed laparoscopically and examined histologically. Histopathologic examination noted all specimens as lymph nodes with no toxic effects of PBV dye. CONCLUSION: We demonstrated that spermatic funicular injection of PVB dye is an effective and accurate method for retroperitoneal lymph node visualization in pigs. The use of this technique in combination with a laparoscopic approach makes RPLND easier and more effective. PMID- 18429687 TI - Case report: the danger of postchemotherapy laparoscopic retroperitoneal lymph node dissection for nonseminomatous testicular carcinoma. AB - Laparoscopic retroperitoneal lymph node dissection (RPLND) is a feasible, minimally invasive procedure for the treatment of testicular cancer patients who require surgery to address the retroperitoneal lymph nodes. We report a case of retroperitoneal recurrent disease including a port-site metastasis secondary to laparoscopic RPLND. PMID- 18429691 TI - Biosynthesis of the iron-molybdenum cofactor of nitrogenase. AB - The iron-molybdenum cofactor (FeMo-co), located at the active site of the molybdenum nitrogenase, is one of the most complex metal cofactors known to date. During the past several years, an intensive effort has been made to purify the proteins involved in FeMo-co synthesis and incorporation into nitrogenase. This effort is starting to provide insights into the structures of the FeMo-co biosynthetic intermediates and into the biochemical details of FeMo-co synthesis. PMID- 18429692 TI - Predicting the emergence of tick-borne infections based on climatic changes in Korea. AB - Granulocytic anaplasmosis (GA) and monocytic ehrlichiosis (ME) are maintained in wild rodent reservoirs and tick vectors in the Republic of Korea. This study investigated the prevalence of 2 tick-borne pathogens, Anaplasma phagocytophilum and Ehrlichia chaffeensis, in wild rodents and ticks in central Korea to identify any significant associations with existing or changing climatic conditions. Specifically, the goal of this study was to develop simple models for the probability of occurrence of an epidemic of GA or ME as a function of climate in an area in a given year. Climatic data from 2 regions, Munsan and Dongducheon, Gyeonggi, in central Korea (between the Demilitarized Zone and Seoul, latitude between 37 degrees N-38 degrees N and longitude between 127 degrees E-128 degrees E), were analyzed with respect to the prevalence of GA and ME in Paju, Yoncheon, Pocheon, and Dongducheon for the period from 2001 to 2005. Rates of A. phagocytophilum and E. chaffeensis decreased as the total yearly precipitation levels and daily humidity increased, and as the daily mean sunshine hours decreased. Rates of A. phagocytophilum and E. chaffeensis from rodent ticks and rodents increased in the fall season. Linear regression analyses evaluating the numbers of positive samples by sample type found that rodent ticks were 6.64 times more likely to be actively infected with A. phagocytophilum than grass ticks or rodents, though the likelihood of any samples testing positive for this pathogen decreased by 0.17 as the annual mean level of precipitation increased by 1 mm. For E. chaffeensis, rodents were 15.67 times more likely to be infected than ticks. Logistic regression analyses evaluating each sample separately found that the odds of infection with A. phagocytophilum were nearly 5 times greater for rodents than ticks. In these analyses, precipitation was one potential factor to account for the prevalence of tickborne diseases. PMID- 18429693 TI - Land cover, landscape structure, and West Nile virus circulation in southern France. AB - The transmission of West Nile virus (WNV) is strongly influenced by environmental factors. In France, two endemic areas for WNV have been identified: Camargue and Var. The objective of our study was to test whether landscape characteristics could be associated with the risk of endemic circulation of WNV in these two ecologically different areas. Equine serological data collected during outbreaks in Var (2003) and Camargue (2004) were used. Both areas were marked out in cells of 5 km(2), and data were aggregated for each cell. Sixteen cells, classified as "high" level viral circulation, and 28 cells, classified as "low" level viral circulation were used for the analysis. The Corine Land Cover database (European Environment Agency) was used to evaluate, for each geographic cell, the area covered by 12 land cover classes, as well as the value of 5 landscape metrics (patch richness and density, edge density, the Shannon's diversity index, and interspersion and juxtaposition index (IJI). Multivariate linear generalized regression showed that IJI as well as the surface covered by heterogeneous agricultural areas were significantly higher in high level WNV circulation cells than in low level ones (p = 0.01 and 0.05 respectively). Both variables are indicators of a complex spatial biotope configuration that may favor the co existence of competent vectors and reservoir hosts: the structure of the landscape thus appeared as a key element in WNV circulation. An internal validation was performed and the model was used to compute a risk map for the French Mediterranean coast. Cells with a probability > 0.8 of having a high level of viral circulation were found near Aix-en-Provence, Beziers, and Perpignan, areas where no serological study has yet been conducted. Equine cases reported in 2006 were all located in the neighborhood of cells having a > 0.8 probability for high WNV circulation status. PMID- 18429694 TI - Eared dove (Zenaida auriculata, Columbidae) as host for St. Louis encephalitis virus (Flaviviridae, Flavivirus). AB - St. Louis encephalitis virus (SLEV) is an emerging Flavivirus in South American countries. Its ecology and biological transmission cycles are scarcely known. Eared doves (Zenaida auriculata) have frequently been found infected by SLEV, and therefore, could be suspected as SLEV hosts. Thirty post-hatch-year eared doves were subcutaneously inoculated with the genotype V SLEV 78V-6507 viral strain and subsequently bled. No deaths or clinical signs of illness were observed in the inoculated doves. The viremia titers ranged from 2 to 5.5 log(10) plaque-forming units (PFU)/mL during 1-7 days postinoculation (dpi), the highest being observed on the 4th dpi. Mosquitoes were collected using can traps baited with chicken and eared doves for comparison. A total of 2792 mosquitoes belonging to 5 species were collected. Ninety percent of the mosquitoes collected in eared dove-baited can traps were Culex quinquefasciatus. Statistical differences were not observed in either Cx. quinquefasciatus (Chi(2) = 0.86; df = 1; p = 0.354) or in Cx. interfor (Chi(2) = 0.63; df = 1; p = 0.426) mosquitoes collected in both chicken- and eared dove-baited can traps. Considering that eared doves were frequently found naturally infected by SLEV, that they developed viremia titers higher than the minimum infection threshold needed to infect Cx. quinquefasciatus, and that these mosquitoes also fed on eared doves, they could be considered competent hosts for SLEV. PMID- 18429695 TI - Naturally infected Lutzomyia sand flies in a Leishmania-endemic area of Brazil. AB - In Brazil, Leishmania transmission involves several species of phlebotomine sand flies that are closely associated with different parasites and reservoirs, giving rise to different transmission cycles. The present study focused on naturally infected phlebotomines originating from Santa Luzia, a municipality near Belo Horizonte, capital of the Brazilian state of Minas Gerais, in which leishmaniasis are endemic. Systematic and non systematic approaches,involving the use of light traps and direct aspiration from resting sites, respectively, were used to collect females and flies. Identification of the captured insects and determination of natural infection by Leishmania spp. were performed using both conventional dissection methods and polymerase chain reaction (PCR). The dissection of 102 sand flies allowed five species of Lutzomyia to be identified, although no flagellate parasite forms were observed.In addition, 211 sand flies were identified, were separated according to species, and were combined into 11 pools of up to 20 individuals each. PCR analyses showed that two of these pools were infected with Leishmania:one pool of Lu. whitmani was infected with Le. (Viannia) spp. and another of Lu. cortelezzii was infected with Le. chagasi. This suggests that Lu. whitmani may be a possible vector of Leishmania in the study area, and more work needs to be performed to assess the role of Lu. cortelezzii as a vector. PMID- 18429696 TI - Experimental and field studies on the suitability of raccoons (Procyon lotor) as hosts for tick-borne pathogens. AB - We investigated the experimental susceptibility and natural exposure of raccoons (Procyon lotor) to five tick-borne pathogens of human and veterinary importance, Ehrlichia canis, E. chaffeensis, E. ewingii, Anaplasma phagocytophilum (ApVariant 1 and Ap-ha HGE-1 strains), and Borrelia lonestari. Infections were assessed by polymerase chain reaction (PCR), indirect fluorescent antibody (IFA) testing, and/or culture isolation methods for at least 30 days postinoculation (DPI). Two E. chaffeensis-inoculated raccoons seroconverted and were transiently PCR positive. One raccoon was culture positive. Laboratory raised Amblyomma americanum nymphs fed on a third infected raccoon failed to become infected. Two A. phagocytophilum (HGE-1)-inoculated raccoons became PCR positive and seroconverted. Both remained positive for at least 74 DPI. In contrast, raccoons inoculated with A. phagocytophilum (Ap-Variant 1) were only transiently PCR positive and only seroconverted with low titers. No evidence of infection was observed for E. ewingii- and B. lonestari-inoculated raccoons. Only one E. canis inoculated raccoon was PCR positive 3 DPI. Serologic testing of wild raccoons from five populations (3 infested with ticks) in Georgia and Florida showed antibodies reactive with E. chaffeensis in the 3 tick-infested populations (range of 30%-46%), E. canis in the same three populations (8%-23%), A. phagocytophilum in a single raccoon from Florida (12%), and Borrelia spp. in all 5 populations (8%-53%). All raccoons were PCR negative for tick-borne pathogens. These data suggest that raccoons are likely not important reservoirs of E. canis, E. ewingii, or B. lonestari. However, raccoons are experimentally susceptible and naturally exposed to E. chaffeensis, and these data support the previous finding that raccoons may be involved in the natural history of A. phagocytophilum. PMID- 18429697 TI - On becoming a plant pathologist: the changing scene. PMID- 18429698 TI - Regulation of food intake through hypothalamic signaling networks involving mTOR. AB - To maintain normal activity, single cells must assure that their energy needs and utilization are continuously matched. Likewise, multicellular organisms must constantly coordinate energy intake and expenditure to maintain energy homeostasis. The brain, and the hypothalamus in particular, plays a critical role in integrating and coordinating several types of signals, including hormones and nutrients, to guarantee such homeostasis. Like single cells, the hypothalamus also profits from intracellular pathways known to work as fuel sensors to maintain energy balance. One such pathway is the mammalian target of rapamycin (mTOR). mTOR integrates different sensory inputs to regulate protein synthesis rates in individual cells, and it has recently been implicated in the central nervous system to regulate food intake and body weight as well. This review provides an overview of the role of hypothalamic intracellular fuel sensors in the overall control of energy balance and discusses the potential contribution of these fuel-sensing mechanisms to the metabolic dysregulation associated with obesity. PMID- 18429699 TI - Nicotinic acid, nicotinamide, and nicotinamide riboside: a molecular evaluation of NAD+ precursor vitamins in human nutrition. AB - Although baseline requirements for nicotinamide adenine dinucleotide (NAD+) synthesis can be met either with dietary tryptophan or with less than 20 mg of daily niacin, which consists of nicotinic acid and/or nicotinamide, there is growing evidence that substantially greater rates of NAD+ synthesis may be beneficial to protect against neurological degeneration, Candida glabrata infection, and possibly to enhance reverse cholesterol transport. The distinct and tissue-specific biosynthetic and/or ligand activities of tryptophan, nicotinic acid, nicotinamide, and the newly identified NAD+ precursor, nicotinamide riboside, reviewed herein, are responsible for vitamin-specific effects and side effects. Because current data suggest that nicotinamide riboside may be the only vitamin precursor that supports neuronal NAD+ synthesis, we present prospects for human nicotinamide riboside supplementation and propose areas for future research. PMID- 18429700 TI - A retrospective managed care claims data analysis of medication adherence to vaginal estrogen therapy: implications for clinical practice. AB - OBJECTIVES: This study sought to assess refill-based treatment duration and adherence to vaginal estrogen therapy (VET) in clinical practice and to compare treatment duration in women prescribed vaginal tablets (VT) or vaginal creams (VC) in clinical trials with that in clinical practice. METHODS: Adults initiating VET between January and June 2004 were identified in 57 commercial health plans in the United States and followed for up to 10 months after treatment initiation. Treatment duration (Kaplan-Meier analysis) and adherence (medication possession ratio [MPR]) were examined for VC and VT cohorts and compared with a weighted average of treatment duration calculated for seven clinical trials identified in the literature. RESULTS: Of 13,074 women (mean age 54 +/- 9.1 years) identified, patients on VT had significantly longer treatment duration compared with patients on VC (149.1 +/- 101.1 days vs. 91.6 +/- 30.0 days, p < 0.01). Among 5,599 patients receiving multiple prescriptions, higher treatment durations were observed for both VT-treated and VC-treated patients (198.5 +/- 82.4 days vs. 177.1 +/- 86.7 days, p < 0.01) compared with 165 days observed in clinical trials. Medication adherence (MPR > or = 80%) was significantly higher among VT users compared with VC users (74% +/- 27% vs. 52% +/- 32%], p < 0.01); OR, 3.24, 95% CI 2.84-3.70). CONCLUSIONS: Duration of VET among patients receiving multiple prescriptions was longer in real-world clinical practice than in clinical trials. Newly treated patients initiating VT were more likely to continue treatment for longer periods and exhibited greater medication adherence than did those on VC. PMID- 18429701 TI - Primary care program directors' perceptions of women's health education: a gap in graduate medical education persists. AB - BACKGROUND: Previous studies found that internal medicine residents are not adequately prepared to provide comprehensive primary care to women. The impact of subsequent national guidelines emphasizing women's health education during residency is unknown. METHODS: We conducted a cross-sectional survey of primary care internal medicine residency program directors (PDs) in the United States. We asked the directors to provide information about themselves and their programs, to rate how strongly they agreed that residents should master each of 13 women's health competencies, to estimate the proportion of their residents who actually master each competency by the end of their residency, and to indicate means by which each competency was taught (articles, lectures, patient care, specialty clinic, other). RESULTS: Of 69 directors contacted, 42 (61%) responded. Most respondents agreed that residents should master all 13 competencies. However, there were significant discrepancies (p < 0.001) between the proportion of respondents who believed their residents should master competencies concerning 10 of 13 women's health topics and the proportion who believed their residents actually did master them. More than one third of PDs estimated that the majority of their residents would not gain the knowledge to diagnose, treat, or counsel women with incontinence, vaginitis, domestic violence, preconception planning, or birth control needs by the end of residency training. Of 18 potential predictors of quality education, only 2 proved significant: number of years the respondent served as program director (p = 0.02) and number of competencies taught by lecture (p = 0.007). CONCLUSIONS: Despite national guidelines endorsing women's health education, a large discrepancy persists between what PDs believe their residents should master and what they estimate their residents actually master. This study suggests a need for substantial improvements in internal medicine residency training to adequately prepare residents to care for women. PMID- 18429702 TI - Fluorescence imaging of membrane dynamics. AB - Imaging membrane dynamics is an important goal, motivated by the abundance of biochemical and biophysical events that are orchestrated at, or by, cellular membranes. The short length scales, fast timescales, and environmental requirements of membrane phenomena present challenges to imaging experiments. Several technical advances offer means to overcome these challenges, and we describe here three powerful techniques applicable to membrane imaging: total internal reflection fluorescence (TIRF) microscopy, fluorescence interference contrast (FLIC) microscopy, and fluorescence correlation spectroscopy (FCS). For each, we discuss the physics underpinning the approach, its practical implementation, and recent examples highlighting its achievements in exploring the membrane environment. PMID- 18429703 TI - Psychophysical evaluation for visual prosthesis. AB - Vision restoration through retinal, optic nerve, and cortical implants is no longer just the stuff of fantasy. The design and development of visual prostheses rapidly move from the engineering phase toward preclinical and clinical trials, yet the benchmarks to determine their efficacy in blind research subjects have received very little attention, and likewise the selection criteria and preparation of early recipients of these devices. This article examines the aspects of vision for which prostheses may be of help, the selection of early prosthesis wearers, and the pre- and postimplant evaluations required to assess safety and efficacy. I concentrate on the functional assessment, and particularly on psychophysical methodology, but also address other measures of safety and efficacy, as well as approaches to vision rehabilitation with visual prostheses. Finally, I speculate what roles the first few generations of visual prostheses may play, and emphasize the importance of using simulations to support the development and rehabilitation process. PMID- 18429704 TI - Neural stimulation and recording electrodes. AB - Electrical stimulation of nerve tissue and recording of neural electrical activity are the basis of emerging prostheses and treatments for spinal cord injury, stroke, sensory deficits, and neurological disorders. An understanding of the electrochemical mechanisms underlying the behavior of neural stimulation and recording electrodes is important for the development of chronically implanted devices, particularly those employing large numbers of microelectrodes. For stimulation, materials that support charge injection by capacitive and faradaic mechanisms are available. These include titanium nitride, platinum, and iridium oxide, each with certain advantages and limitations. The use of charge-balanced waveforms and maximum electrochemical potential excursions as criteria for reversible charge injection with these electrode materials are described and critiqued. Techniques for characterizing electrochemical properties relevant to stimulation and recording are described with examples of differences in the in vitro and in vivo response of electrodes. PMID- 18429705 TI - An end to suppressing public health information. PMID- 18429706 TI - Another inquiry into public hospitals? PMID- 18429707 TI - Ensuring the safety of new medications and devices: are naltrexone implants safe? PMID- 18429708 TI - Unplanned admissions to two Sydney public hospitals after naltrexone implants. AB - OBJECTIVE: To describe hospital presentations related to the use of naltrexone implants, an unlicensed product used in Australia for treating heroin dependence. DESIGN: Retrospective case file audit. SETTING: Two Sydney teaching hospitals. PATIENTS: Identified through referrals to Drug and Alcohol Consultation-Liaison services over a 12-month period, August 2006 to July 2007. MAIN OUTCOME MEASURES: Diagnosis, management and duration of admission. RESULTS: Twelve cases were identified: eight were definitely or probably related to naltrexone implants or the implantation procedure (rapid detoxification). Of these, six patients had severe opiate withdrawal and dehydration, with an average hospital stay of 2.3 days. One patient had an infection at the implant site, and one an underlying anxiety disorder requiring psychiatric admission. Three patients had analgesia complications, and one had unrelated cardiac arrhythmia. CONCLUSIONS: These severe adverse events challenge the notion that naltrexone implants are a safe procedure and suggest a need for careful case selection and clinical management, and for closer regulatory monitoring to protect this marginalised and vulnerable population. PMID- 18429709 TI - Metformin and lactic acidosis in an Australian community setting: the Fremantle Diabetes Study. AB - OBJECTIVE: To determine the incidence of lactic acidosis in community-based patients with type 2 diabetes, with special reference to metformin therapy. DESIGN: Substudy within a longitudinal observational study, the Fremantle Diabetes Study (FDS). PARTICIPANTS AND SETTING: 1279 patients from a postcode defined population of 120 097 people in Western Australia. MAIN OUTCOME MEASURES: Confirmed hospitalisation with lactic acidosis identified through the WA Data Linkage System during two periods: (1) from study entry, between 1993 and 1996, and study close in November 2001; and (2) from study entry to 30 June 2006. RESULTS: At entry, 33.3% of patients were metformin-treated, and 23.1% of these had one or more contraindications to metformin (55.1% and 38.0%, respectively, after 5 years' follow-up). Five confirmed cases of lactic acidosis were identified during 12 466 patient-years of observation; all had at least one other potential cause, such as cardiogenic shock or renal failure. From study entry to close, the incidence was 0/100,000 patient-years in both metformin-treated and non-metformin-treated patients. Between study entry and 30 June 2006, incidence was 57/100,000 patient-years (95% CI, 12-168) in metformin-treated patients and 28/100,000 patient-years (95% CI, 3-100) in the non-metformin-treated group, an incidence rate difference of -30 (-105 to 46) (P=0.4). CONCLUSION: The incidence of lactic acidosis in patients with type 2 diabetes is low but increases with age and duration of diabetes, as cardiovascular and renal causes become more prevalent. Metformin does not increase the risk of lactic acidosis, even when other recognised precipitants are present. PMID- 18429710 TI - Impact of specialty on attitudes of Australian medical practitioners to end-of life decisions. AB - OBJECTIVE: To compare attitudes and practices of Australian medical practitioners, by specialty, to a range of medical decisions at the end of life. DESIGN, SETTING AND PARTICIPANTS: As part of an international study, in 2003, a structured questionnaire was mailed to 2964 medical practitioners drawn from membership registers of Australian and Australasian professional colleges. Data from 1478 questionnaires were statistically analysed using validated instruments. MAIN OUTCOME MEASURES: Practitioners' willingness to comply with requests from patients and/or their relatives for symptom relief which might also hasten death; provision of terminal sedation and euthanasia, or willingness to provide these on their own initiative. RESULTS: Respondents reported being much more willing to comply with a patient's request for increasing symptom relief, even at risk of hastening death, than for terminal sedation. Over a quarter of respondents would provide terminal sedation to competent patients on their own initiative. A small number of respondents would intentionally hasten death. There were significant differences by specialty for all three actions. Oncologists, palliative care physicians and geriatricians were least likely to actively hasten death, and more likely to act unilaterally to relieve symptoms as a medical necessity. CONCLUSIONS: Perceptions about the causation of death and aspects of medical culture appear to influence physicians' attitudes towards medical decisions at the end of life. Our findings have implications for medical education, interprofessional communication and discussion between the medical profession and the community. PMID- 18429711 TI - Concordance between real-time telemedicine assessments and face-to-face consultations in paediatric otolaryngology. AB - OBJECTIVE: To determine agreement between diagnoses and management plans made during an initial videoconference appointment and subsequent face-to-face consultations in paediatric ear, nose and throat (ENT) surgery. DESIGN AND SETTING AND PARTICIPANTS: A paediatric ENT clinic servicing patients from Bundaberg, Queensland, was conducted through the Centre for Online Health at the Royal Children's Hospital (RCH) in Brisbane. Between January 2004 and February 2006, 152 consultations with 97 patients were carried out. We retrospectively audited patients' charts to compare the diagnosis and management plan formulated at the initial videoconference and the eventual diagnosis and surgical management after face-to-face consultation. The clinical outcomes for children who were not recommended for surgery at the RCH were ascertained by telephone survey. MAIN OUTCOME MEASURES: Agreement between videoconference and face-to-face consultation findings. RESULTS: Of the 97 patients, 75 were recommended for surgical management at the RCH. The remaining patients were either referred back to their general practitioner (9), followed up by the regional paediatrician (10) or lost to follow-up (3). At the conclusion of the study, seven patients were still awaiting surgery and were excluded. Among the 68 patients seen via videoconference and in person, the recorded diagnosis was the same in 99% of cases (67). Surgical management decisions were the same in 93% of cases (63). Telephone follow-up with paediatricians and GPs confirmed that there were no missed diagnoses or ongoing ENT-related problems in the 19 patients referred back to their care. CONCLUSIONS: Decisions about ENT surgical interventions for children assessed during videoconference clinics are in close agreement with decisions made by the same surgeon at face-to-face consultation. The way is open to employ telemedicine more widely for pre-admission ENT assessment. However, as in any telemedicine work, widespread application requires care. PMID- 18429712 TI - Changes to the University of Sydney medical curriculum. AB - In 1997, the University of Sydney replaced its undergraduate medical course with a new 4-year graduate medical program. The course was reaccredited by the Australian Medical Council (AMC) in 2005. Suggestions for further development by the AMC led to a comprehensive review of the course, commencing in July 2006. This article describes the review process and outlines key recommendations that came out of the review that have been or are being implemented. PMID- 18429713 TI - Nationally consistent assessment of international medical graduates. AB - The need for consistency in assessing international medical graduates for work in Australia led the Council of Australian Governments, in 2006, to direct health ministers to implement a nationally consistent approach. An Implementation Committee was established in late 2006 to oversee the development of the new assessment process; the first steps were completed by July 2007 and further development will occur over the next 12 months. The pre-existing Australian Medical Council (AMC) examination pathway will continue to be available, and there will be two additional pathways for non-specialists. The pre-existing pathway for specialists is being revised. Elements that are being introduced include: standardised pre-employment assessment, including an off-shore screening examination; assessment of competence against a standardised position description and, if necessary, a structured clinical interview by an AMC-accredited provider before obtaining limited registration; orientation to the job, the Australian health care system and to communication and cultural issues; standardised supervision and supervisory reporting; a requirement for compulsory continuing professional development for reregistration; workplace-based assessment; assessment by an AMC-accredited provider before gaining full registration; and consistency of assessment for specialists by specialist colleges. Some elements have been operating in some jurisdictions from July 2007, and there is a commitment to full implementation in participating jurisdictions by July 2008. PMID- 18429715 TI - The Royal North Shore Hospital inquiry: an analysis of the recommendations and the implications for quality and safety in Australian public hospitals. AB - In October 2007, the New South Wales Parliament appointed a Joint Select Committee to inquire into the quality of patient care at Royal North Shore Hospital (RNSH). The inquiry was initiated in response to the publicity and complaints surrounding a patient who had a miscarriage in the toilets of the RNSH emergency department waiting area. The Committee held four public hearings and received 103 submissions. It handed down 45 recommendations in its report on 20 December 2007. There has been criticism from clinicians and others that the recommendations are too general and will not effect significant change for the severe systemic problems affecting the hospital. This article represents the view of some of the clinicians who work at RNSH, and who gave evidence at the inquiry, on the recommendations and some possible solutions for the health system in general. PMID- 18429717 TI - The way we treat each other. AB - There is a heavy burden of disease associated with family violence, discrimination, bullying and social exclusion. These important causes of suffering and loss of productivity all relate to a very fundamental feature of human existence and civil(ised) societies--the way we treat each other. We can, and do, make each other sick. Reducing the resultant human and economic costs has major implications for the way we distribute opportunity, wealth and amenity. These, in turn, have implications for the way we protect and empower minority groups, and for legislation, education and the availability of, and access to, services. PMID- 18429716 TI - Current use of Australian snake antivenoms and frequency of immediate-type hypersensitivity reactions and anaphylaxis. AB - OBJECTIVE: To investigate current use of Australian snake antivenoms and the frequency and severity of immediate-type hypersensitivity reactions. DESIGN: Nested prospective cohort study as part of the Australian Snakebite Project. PATIENTS AND SETTING: Patients receiving snake antivenom in Australian hospitals between 1 January 2002 and 30 November 2007. MAIN OUTCOME MEASURES: The use of CSL Limited antivenom; frequency and severity of hypersensitivity reactions to antivenom; premedication and treatment of these reactions. RESULTS: Snake antivenom was administered to 195 patients, mostly for venom-induced consumption coagulopathy (145 patients, 74%), followed by non-specific systemic effects (12%), neurotoxicity (5%) and myotoxicity (4%). Antivenom was given to nine patients (5%) without evidence of envenoming or who were bitten by a species of snake for which antivenom is not required. The commonest antivenoms used were brown snake (46%), tiger snake (30%) and polyvalent (11%). The median dose was four vials (interquartile range, 2-5 vials), and 24 patients received two different types of antivenom. Immediate-type hypersensitivity reactions occurred in 48 patients (25%); 21 satisfied our definition of anaphylaxis, with 11 moderate and 10 severe cases, including nine in which patients were hypotensive. The remaining 27 reactions were mild (skin only). Adrenaline was used in 26 cases with good effect. The frequency of reactions to tiger snake (41%) and polyvalent (41%) antivenoms was higher than that to brown snake antivenom (10%). Hypersensitivity reactions occurred in 11 of 40 patients receiving any form of premedication (28%) and in 2 of 11 given adrenaline for premedication (18%) versus 20 of 86 not receiving premedication (23%). CONCLUSIONS: Antivenom was used appropriately, and most commonly for coagulopathy. Hypersensitivity reactions were common, but most were not severe. The discretionary use of premedication was not associated with any reduction in reactions. PMID- 18429720 TI - Bystander basic life support: an important link in the chain of survival for children suffering a drowning or near-drowning episode. AB - Eight children suffered drowning or near-drowning in Sydney pools over an 11-day period in January 2007. Four received basic life support (BLS) within 5 minutes of immersion and survived with good functional neurological outcomes. The other four were not discovered for >or= 5 minutes and all died. This cluster serves as a reminder that timely effective bystander BLS is crucial to survival and good clinical outcomes in near-drowning episodes. PMID- 18429721 TI - Spontaneous intracranial hypotension. PMID- 18429722 TI - Tissue plasminogen activator for acute ischaemic stroke. PMID- 18429725 TI - Tissue plasminogen activator for acute ischaemic stroke. PMID- 18429726 TI - Tissue plasminogen activator for acute ischaemic stroke. PMID- 18429728 TI - Early intervention in youth mental health. PMID- 18429729 TI - Sternal fracture in an Australian Rules footballer. PMID- 18429730 TI - Why are community psychiatric services in Australia doing it so hard? PMID- 18429731 TI - Intervening early to reduce developmentally harmful substance use among youth populations. PMID- 18429732 TI - Barrett's oesophagus and columnar metaplasia: saying what we mean. PMID- 18429733 TI - Health technology assessment in Australia. PMID- 18429734 TI - The Australian Primary Health Care Research Institute: rising to the challenge of applying knowledge from research to Australian policy. PMID- 18429735 TI - Bringing evidence to bear on policy processes: the challenge of the Australian Primary Health Care Research Institute. PMID- 18429736 TI - Models of chronic disease management in primary care for patients with mild-to moderate asthma or COPD: a narrative review. AB - OBJECTIVE: To review the literature for any promising strategies for the primary care management of mild-to-moderate asthma and chronic obstructive pulmonary disease (COPD) in adults. METHODS: Using "MeSH" terms for COPD, asthma and primary health care, we conducted an extensive literature search for relevant meta-analyses, systematic reviews, narrative reviews, reports and individual studies. Grey literature was also included. We chose a narrative review approach because of substantial heterogeneity of study designs in the literature. RESULTS: 1119 articles of potential relevance were retained, of which 246 were included in our review. There was insufficient evidence to determine whether general practitioners with a special interest (GPwSI) in respiratory care improved the diagnosis and management of mild-to-moderate COPD. An asthma service involving GPwSI increased respiratory drug costs but reduced the costs for less specific drugs. No clear benefit has been shown for practice nurse-run asthma clinics in primary care compared with usual care in altering asthma morbidity, quality of life, lung function or medication use. Evidence to determine the effectiveness of practice nurse-run COPD clinics could not be found. Self-management education, GP review and action plans may produce short-term benefits for asthma patients, particularly those with moderate-to-severe disease, but the evidence for a similar approach to patients with mild-to-moderate COPD is equivocal. There has been poor uptake of respiratory clinical guidelines relevant to primary care - partly because most guidelines are based on moderate-to-severe disease. Spirometry programs in primary care are useful for differential diagnosis of asthma and COPD. Spirometry may alter the management of mild asthma, but there is a lack of evidence that it alters the management of COPD in primary care. CONCLUSION: The role of primary health care in management of mild-to-moderate asthma and COPD requires further investigation using randomised controlled trials. PMID- 18429737 TI - Chronic disease management in primary care: from evidence to policy. AB - OBJECTIVES: To review the effectiveness of chronic disease management interventions for physical health problems in the primary care setting, and to identify policy options for implementing successful interventions in Australian primary care. METHODS: We conducted a systematic review with qualitative data synthesis, using the Chronic Care Model as a framework for analysis between January 1990 and February 2006. Interventions were classified according to which elements were addressed: community resources, health care organisation, self management support, delivery system design, decision support and/or clinical information systems. Our major findings were discussed with policymakers and key stakeholders in relation to current and emerging health policy in Australia. RESULTS: The interventions most likely to be effective in the context of Australian primary care were engaging primary care in self-management support through education and training for general practitioners and practice nurses, and including self-management support in care plans linked to multidisciplinary team support. The current Practice Incentives Payment and Service Incentives Payment programs could be improved and simplified to encourage guideline-based chronic disease management, integrating incentives so that individual patients are not managed as if they had a series of separate chronic diseases. The use of chronic disease registers should be extended across a range of chronic illnesses and used to facilitate audit for quality improvement. Training should focus on clear roles and responsibilities of the team members. CONCLUSION: The Chronic Care Model provides a useful framework for understanding the impact of chronic disease management interventions and highlights the gaps in evidence. Consultation with stakeholders and policymakers is valuable in shaping policy options to support the implementation of the National Chronic Disease Strategy in primary care. PMID- 18429738 TI - Seriously working together: integrated governance models to achieve sustainable partnerships between health care organisations. AB - OBJECTIVE: To identify sustainable governance arrangements for health care organisations undertaking integrated health service delivery based on best available evidence. METHOD: Systematic review of the literature (1990-2006), supported by key informant interviews as an integrative process. RESULTS: 16 studies met our selection criteria. All described enablers of and barriers to delivering integrated health services. We identified three models for integrated health care governance with a demonstrated ability to be sustained in the medium term. Common themes that emerged as the logical starting point for more ambitious integrated governance arrangements regionally were: the need for a clear separation between governance and operational management; and the need for local communities with the vision, leadership and commitment to extend health service integration. These themes were reinforced by interviews with key informants. Careful measurement of the process, impact and outcomes of such activities was often overlooked. CONCLUSION: State governments are increasingly attempting to work with non-government organisations and the private sector to maximise scarce resources in the face of increasing health care demand. Ambitious integration agendas must be underpinned by effective governance mechanisms that are appropriate to the undertaking, the stakeholders involved and the scale of delivery. PMID- 18429739 TI - Multidisciplinary care planning and teamwork in primary care. AB - OBJECTIVE: To examine policy and implementation issues around multidisciplinary care planning (MDP) as a means of improving outcomes for patients with chronic disease and/or complex care needs. METHODS: We conducted a series of five systematic reviews of the literature from 1990 to 2006, sampling a spectrum of issues associated with chronic disease and complex health care needs, with a focus on planning and provision of multidisciplinary care. RESULTS: Our review showed that MDP does improve many functional outcomes in the areas studied. Analysis of MDP programs involves examination of two groups of variables - the multidisciplinary components (a range of clinical perspectives and specialist knowledge) and team components (eg, communication and support). Implementing MDP requires changing patterns of interaction between care providers, alignment of roles and work practices, and changes to organisational arrangements. CONCLUSION: While MDP improves many functional outcomes, widespread implementation of MDP in standard practice will require complex and targeted strategies. Devising and testing such strategies is a prerequisite for widespread, routine use of MPD in chronic disease management. PMID- 18429740 TI - Coordinating primary health care: an analysis of the outcomes of a systematic review. AB - OBJECTIVES: To identify the types of strategy used to coordinate care within primary health care (PHC) and between PHC, health services and health-related services in Australia and other countries that have comparable health systems, and to describe what is known about their effectiveness; to review the implications for health policy and practice in Australia. METHODS: We conducted a systematic review of the literature (January 1995 to March 2006) relating to care coordination in Australia, the United States, the United Kingdom, New Zealand, Canada and The Netherlands. Our review was supplemented by consultations with academic experts and policymakers. RESULTS: Six types of strategy were identified at patient/provider level, falling into two groups: (i) communication and support for providers and patients, and (ii) structural arrangements to support coordination. These were broadly consistent with existing typologies. All were associated with improved health and/or patient satisfaction outcomes in more than 50% of studies, and interventions using multiple strategies were more successful than those using single strategies. CONCLUSIONS: The largely incremental approach to improving coordination of care in Australia has involved a broad range of strategy types but has also perpetuated existing structural problems. Reforms in governance, funding and patient registration in primary health care would provide a stronger base for effective care coordination. PMID- 18429741 TI - The implementation and impact of different funding initiatives on access to multidisciplinary primary health care and policy implications. AB - OBJECTIVE: To review the implementation and impact of different funding initiatives across the health systems of three different countries - England, New Zealand and Australia - on the achievement of multidisciplinary primary health care (PHC) and to reflect on policy implications for Australia. METHODS: A systematic review of the literature involving three stages: (i) identification and description of initiatives; (ii) a systematic review of their implementation and impact from 1995 to mid 2006; and (iii) an updated review of published literature from mid 2006 to mid 2007. RESULTS: Few studies employed control groups, and the results should therefore be interpreted with caution. In all three countries, funding has supported general practitioner access to a broad range of providers. In Australia, financial incentives have been the main mechanism for bringing about change, whereas in both England and New Zealand, they are part of a broader range of funding reforms including the introduction of capitation and practice-based commissioning. The lack of patient data makes it difficult to assess the extent to which the Australian financial incentives have generally improved population access to a broader range of PHC providers. CONCLUSION: Individual, patient-level, financial incentives may present significant impediments for population subgroups with complex needs. Alternative funding arrangements, such as capitation and contracting, could be more widely adopted in Australia to enhance access to care for vulnerable population groups without fundamentally changing the overall fee-for-service financing arrangements. PMID- 18429742 TI - Primary care funding and organisational policy options and implications: a narrative review of evidence from five comparator countries. AB - OBJECTIVE: To review innovative models of primary care in different countries in order to identify potential mechanisms for reforming primary care policy in Australia. METHODS: We conducted a narrative review and synthesis of evidence about models of primary care from four English-speaking comparator countries (New Zealand, Canada, the United Kingdom and the United States) and one European country (The Netherlands), with a particular focus on the relevance and applicability of these models to Australia. RESULTS: We identified four key mechanisms for bringing about reform in primary care: flexible funding, quality frameworks, regional-level primary care organisations, and primary care infrastructure. These mechanisms are interdependent. CONCLUSION: There are tensions and tradeoffs involved in balancing professional and bureaucratic control and in linking quality and accountability mechanisms. Enhanced linkage between researchers, policymakers and professional groups could assist in exploring options for effective primary care reform. PMID- 18429743 TI - "Beyond workforce": a systemic solution for health service provision in small rural and remote communities. AB - Successful, "innovative" primary health care (PHC) models exist that have adapted to the specific circumstances of their rural and remote context. A typology of discrete, integrated, comprehensive and outreach rural and remote services exists rather than a "one coat fits all" PHC health service model. Successful models are characterised by macro-scale environmental enablers (supportive health policy, federal-state relations, and community readiness) and five essential service requirements (workforce organisation and supply; funding; governance, management and leadership; linkages; and infrastructure). Service sustainability depends on ensuring that key systemic service requirements are met at the local level in ways that accord with, and are supported by, the broader macro-scale environmental enablers. Based on these principles, these model types are amenable to generalisation and evaluation in other regions. PMID- 18429744 TI - Depression in primary health care: from evidence to policy. AB - OBJECTIVE: To consider the implications for mental health policy of a recent synthesis of the literature on the effectiveness of different service delivery models for depression in primary care. METHODS: A discussion based on the results of several systematic reviews of primary care models for depression management. Primary care was defined broadly within a prevention, early-intervention, treatment and recovery/support framework, and incorporated both community and general practice settings. RESULTS: There were promising effective models for depression interventions both in the broader community and in general practice settings. CONCLUSIONS: There is a need to support evidence-based models for depression care, including innovative new technologies for facilitating consumer self-management of depression. The ability of practitioner training and guideline implementation to improve consumer outcomes for depression is limited. Policies and incentives are required to facilitate the reorganisation of general practice and, in particular, the implementation of care management as well as enhanced care and guided self-help in these settings. PMID- 18429745 TI - Preventing growth faltering among Australian Indigenous children: implications for policy and practice. AB - OBJECTIVE: To determine what preventive models or programs are most likely to improve patterns of growth faltering in children aged under 5 years in remote Australian Indigenous communities. METHODS: Nine electronic databases and the websites of key stakeholder, government and non-government agencies were searched. Two reviewers independently assessed articles for inclusion and for study quality. All types of study design were eligible. RESULTS: 140 studies assessing a diverse range of interventions were identified. Of these, 51 articles referring to 44 individual programs and 7 review articles met the review criteria. The evidence for the effectiveness of many interventions to prevent growth faltering is not strong, and any observed effects are modest. Community based nutrition education/counselling and multifaceted interventions involving carers, community health workers and community representatives, designed to meet program best-practice requirements and address the underlying causes of growth faltering, may be effective in preventing growth faltering. Other interventions, such as food distribution programs, growth monitoring, micronutrient supplementation and deworming should only be considered in the context of broader primary health care programs and/or when there is an identified local need. CONCLUSION: For remote Indigenous communities, development and implementation of programs should involve a consideration of the evidence for potential impact, strength of community support and local feasibility. Given the lack of strong evidence supporting programs, any new or existing programs require ongoing evaluation and refinement. PMID- 18429746 TI - Review of evidence to guide primary health care policy and practice to prevent childhood obesity. AB - OBJECTIVES: To identify key barriers to effective engagement of primary health care (PHC) providers and families in promoting healthy weight among children aged 2-6 years, and to examine promising interventions to identify policy goals to overcome these barriers. METHODS: We conducted a literature review of published and unpublished articles from January 1990 to February 2006 using keywords relating to childhood obesity, risk factors, prevention, populations and primary care provider interventions, constraints and models. We identified barriers to engagement by PHC providers. Appraisal of "promise" was based on best available evidence and consideration of strengths and weaknesses of interventions in specific contexts and settings. RESULTS: Of 982 interventions aimed at the primary prevention of overweight and obesity among children, few related to 2-6 year-olds, with only 45 interventions meeting the inclusion criteria and 11 ranking highly on key criteria. Areas of weakness were low-level engagement by PHC providers, focus on single risk factors rather than a multidimensional approach, and lack of a population focus. A range of administrative, attitudinal, knowledge, skills and training issues were identified as barriers to effective engagement of different PHC providers with parents and other early childhood service providers. CONCLUSIONS: Engagement of PHC providers in prevention of childhood obesity requires a systematic approach involving practice protocols, assessment tools, client support material and referral pathways, as well as adequate training and sufficient staff for implementation. A more comprehensive approach could be promoted by increased collaboration, agreed role delineation, consistent public health messages and better coordination between PHC providers and other service providers, facilitated at service policy and administration level. PMID- 18429747 TI - The effectiveness of competency-based education in equipping primary health care workers to manage chronic disease in Australian general practice settings. AB - OBJECTIVE: To review the literature on the effectiveness of competency-based education (CBE) as a means of equipping the Australian general practice workforce to deliver optimal chronic disease outcomes to articulate policy options for the Australian context. METHODS: Systematic review of the literature (1991-2005) using a narrative approach followed by analysis of the findings using the actors/context/ processes/content framework of Buse et al. RESULTS: Few high quality studies were identified. National policy options include incorporating clear statements about education and training, research and evaluation in any policy document targeting chronic disease; and provision of funding to enhance general practice teaching facilities and/or facilitate the development of supportive coordinating and administrative structures for training practices. Designers of CBE should consider five key questions: Are the educational objectives of the CBE clearly aligned with the chronic disease or workforce related outcomes of interest? Is the design of the CBE sound? Have similar educational programs targeting the same outcomes been identified and every attempt made to maximise synergies between programs? Are the educational designers fully aware of and working within the existing complexity of the training environment? Are all involved in the program actively managing the process of change? CONCLUSIONS: Policy options range from those relatively simple and achievable to more complex and difficult. The full report is available at http://www.anu.edu.au/aphcri/Domain/Workforce/final_25_glasgow.pdf. PMID- 18429749 TI - [Clinical and economical effectiveness of outpatient diagnostic coronary angiography]. AB - Introduction into clinical practice of transradial coronary angiography (CA) made it possible to conduct the procedure in outpatients and thus increase number of CA and diminish its cost. Aim of the study was to assess possibility, safety and economical efficacy of outpatient CA. Between April 2004 and August 2007 CA was carried out in 133 outpatients without overt heart failure, unstable angina, complex disturbances of cardiac rhythm or conduction. Comparison group comprised 187 patients subjected to CA within framework of short term hospitalization program. There were no complications associated with the use of either transradial or transfemoral approach. Total average cost of outpatient CA was 19% less than that of inhospital CA. Thus outpatient CA with transradial approach appears to be safe and effective procedure with low risk of complications in patients with stable ischemic heart disease. PMID- 18429750 TI - [Excimer laser coronary angioplasty in combination interventional treatment of patients with acute coronary syndrome]. AB - The authors present first experience of application of the system for rheolytic therapy AngioJet in a group of patients with acute myocardial infarction. They describe principle of the work of rheolytic systems, possible procedure related complications. They also discuss results of clinical studies analyzing efficacy of antithrombotic therapy, efficacy and safety of rheolytic therapy in patients with acute coronary syndrome and complicated atherosclerotic plaques in coronary arteries. PMID- 18429748 TI - Enzymes of glycerol and glyceraldehyde metabolism in mouse liver: effects of caloric restriction and age on activities. AB - The influence of caloric restriction on hepatic glyceraldehyde- and glycerol metabolizing enzyme activities of young and old mice were studied. Glycerol kinase and cytoplasmic glycerol-3-phosphate dehydrogenase activities were increased in both young and old CR (calorie-restricted) mice when compared with controls, whereas triokinase increased only in old CR mice. Aldehyde dehydrogenase and aldehyde reductase activities in both young and old CR mice were unchanged by caloric restriction. Mitochondrial glycerol-3-phosphate dehydrogenase showed a trend towards an increased activity in old CR mice, whereas a trend towards a decreased activity in alcohol dehydrogenase was observed in both young and old CR mice. Serum glycerol levels decreased in young and old CR mice. Therefore increases in glycerol kinase and glycerol-3-phosphate dehydrogenase were associated with a decrease in fasting blood glycerol levels in CR animals. A prominent role for triokinase in glyceraldehyde metabolism with CR was also observed. The results indicate that long-term caloric restriction induces sustained increases in the capacity for gluconeogenesis from glycerol. PMID- 18429751 TI - [Clinical efficacy of ivabradine in patients with ischemic heart disease in combination with chronic obstructive pulmonary disease]. PMID- 18429752 TI - [Genetic aspects of individual sensitivity to betaxolol in patients with arterial hypertension]. AB - Association of polymorphism of b1-adrenoreceptors gene and cytochrome 2D6 gene with efficacy of b-adrenoblocker betaxolol was studied in 81 patients with I and II degree arterial hypertension. Betaxolol (10-20 mg/day) was given for 4 weeks, its efficacy was assessed by office blood pressure (BP) measurements, 24-hour BP and ECG monitoring and standard exercise test. At the end of the study significant lowering of systolic and diastolic BP was noted by 11,8 +/- 2,47 (p=0,001) and 7,8 +/- 1,68 mm Hg (p=0,001), respectively. Heart rate (HR) at rest lowered by 19,8 +/- 1,96 beats/min (p=0,0001). At analysis of individual reaction of patients to treatment with betaxolol it turned out that decrease of BP and HR was variable, but their distribution in the group did not differ significantly from normal. Hypotensive activity and influence on HR were confirmed by results of all instrumental investigations. No significant differences were revealed in dynamics of systolic and diastolic BP both at rest and at effort between patients with different genotypes of polymorphic marker Gly389Arg of ADRB1 gene. Compared with carriers of genotype Ser/Ser carriers of genotype Ser/Pro of polymorphic marker Pro34Ser of Cyp2D6 gene had significantly more pronounced decrease of HR at the background of treatment with betaxolol: - 32,6 +/- 4,77 and - 18,4 +/- 2,01 beats/min (p=0,023) at rest and - 30,1 +/- 3,05 and - 24,0 +/- 2,59 beats/min (p=0,043) at maximal exercise, respectively. These patients had also more pronounced lowering of diastolic BP at maximal work load and more pronounced increase of exercise duration at the background of treatment. Thus efficacy of betaxolol in patients with hypertension was associated solely with genotype of polymorphic marker Ser34Pro of CYP2D6 gene. In patients having in genotype Pro allele of polymorphic marker Pro34Ser of CYP2D6 gene therapy with betaxolol is more effective, than in homozygote carriers of Ser allele. This can be related to low rate of metabolism of the preparation in these patients. PMID- 18429753 TI - [Influence of polymorphism's of endothelial nitric oxide synthase gene and polymorphism of NADPH oxidase gene on development of complications of arterial hypertension]. AB - The aim of the study was to analyze the prevalence of polymorphism Glu298Asp of endothelial nitric oxide synthase gene and C242T p22 phox polymorphism of NADPH oxidase gene in patients with arterial hypertension (AH) and their influence on AH complications. The study included 272 AH patients, average age 50,7 years. The following analyses were performed: clinical analysis of the blood, general analysis of the urine, lipid spectrum, plasma electrolytes, creatinine, glucose, electrocardiography, echocardioscopy, examination of eye vessels, ultrasound examination of the carotid arteries, determination of microalbuminuria. The polymorphism Glu298Asp of endothelial nitric oxide synthase gene and C242T p22 phox polymorphism of NADPH oxidase gene were detected with two methods: polymerase chain reaction and restrictase reaction. The control group for Glu298Asp polymorphism detection included 102 healthy Russian donors aged 18 to 50 years. Genotypes prevalence in AH patients was as follows: GG 58,8%, GA 32,3%, AA 8,9%, and CC 48,2%, CT 44,9%, TT 6.9%. In the control group: GG 53%, GA 36%, AA 11% and CC 42%, CT 54%, TT 4%. These polymorphisms did not affect the incidence of complications, such as obliterating atherosclerosis of the lower extremity vessels, ischemic heart disease, and acute insufficiency of cerebral circulation, chronic heart failure, left ventricular hypertrophy, microalbuminuria, carotid arteries atherosclerosis. PMID- 18429754 TI - [Clinical assessment of metabolic effects of antihypertensive therapy]. PMID- 18429755 TI - [Left ventricular hypertrophy and thickening of common carotid artery wall in men aged 40 - 54 years with I degree arterial hypertension]. AB - We carried out clinical-instrumental examination of 456 men aged 40-54 years. First degree arterial hypertension was revealed in 165 men. Left ventricular hypertrophy was found in 48 (30%), increased intima-media thickness (IMT) - in 67 (41%) patients. There was significant medium power relationship between IMT and left ventricular myocardial mass (correlation coefficient 0.41). Formation of left ventricular hypertrophy was related to parameters of 24 hour blood pressure monitoring, arterial hypertension in brothers and sisters, body weight, and duration of obesity. Significant medium power relation was obtained between tension of endothelial system of hemostasis (protein C) and severity of left ventricular hypertrophy (correlation coefficient - 0,3). Age, heredity, low density lipoprotein, cholesterol, uric acid level mattered for IMT increase. PMID- 18429756 TI - [Effect of antihypertensive agents from various pharmacological groups on blood pressure reaction during stress-testing. Part I. Comparative characteristics of medications, exerting effect of sympathoadrenal block]. PMID- 18429757 TI - [Cytochrome P4502C9(CYP2C9) gene polymorphism and safety of therapy with warfarin]. AB - Aim of the study was to investigate frequency and influence of alleles CYP2C9*2 and CYP2C9*3 on pharmacokinetics, pharmacodynamics and dosing regimen of warfarin and on development of hemorrhagic complications. We included 84 patients (mean age 62,8 +/- 10,5 years). Duration of follow-up varied between 1 month and 1 year. Carriage of allele variants was determined by polymerase chain reaction, measurement of plasma wafarin concentration was carried out with the help of high performance liquid chromatography. Wild type (CYP2C9*1/*1) was found in 68% of patients; overall frequency of 2C9*1/*1, *l/*3, *2/*2, *3/*3, *2/*3 genotypes was 32%. Average maintenance doses of warfarin for patients with allele variants CYP 2C9 *2 and 2C9 *3 were 3.6 and 3.1 mg/day, respectively, what was significantly lower than in wild type homozygotes (6.1 mg/day). Wild type homozygotes (1) had the highest warfarin clearance (3,51 ml/min). In carriers of 2C9 *2(2) and 2C9 *3(3) warfarin clearance was significantly lower (2.42 and 1.82 ml/min; p1 - 2 = 0,05; p1 - 3 = 0,0008). In carriers of allele variants CYP2C9*2, CYP2C9*3 values of international normalized ratio > 3,0 were met more often, especially in carriers of CYP2C9*3 (in 100% of cases) vs. 28% in wild type homozygotes (p=0,02). Carriers of CYP2C9*3 compared with wild type homozygotes had more hemorrhagic complications (67% and 16%, respectively, p=0,0008). Thus cytochrome P450 2C9 gene polymorphism influences frequency of development of hemorrhagic complications, metabolic clearance, and magnitude of warfarin maintenance dose. PMID- 18429758 TI - [Efficacy and safety of the use of metformin in patients with chronic heart failure and type 2 diabetes mellitus. results of the study "rational effective mulicomponent therapy in the battle against diabetes mellitus in patients with chronic heart failure"]. AB - Aim of the investigation was to study safety of therapy with metformin and its effect on clinical, hemodynamic, functional and neurohumoral status in patients with chronic heart failure and type 2 diabetes mellitus DM). Eighty one patients with light and moderate NYHA functional class (FC) II-III CHF, left ventricular ejection fraction < 45%, and DM were examined. As a result of randomization 2 groups were formed: with active (n=41) and usual (n=40) treatment. In active group with achievement of target levels of glycemia 24 (59%) patients were on oral hypoglicemic drags, 17 (41) patients received. All patients were on basal therapy of CHF. Initially efficacy and safety of metformin was investigated in a cohort of active treatment (jn metformin n=29, control n=12), including patients who were prescribed metformin not for the whole period. In addition in active group analysis was carried out among patients, who continually were treated with metformin for 12 months (n=30) in comparison with patients never treated with metformin (n=8). Total duration of the period of treatment and supervision was 12 months. Control examination was conducted before randomization, after 6 months of treatment, at the end of the study and included assessment of clunico-functional status of patients, renal function (GFR), neurohumoral profile (MNUP, NA, AII). The state of carbohydrate metabolism was assessed with the help of determination of HBA1C level and test with nutritional load given as of common breakfast -- 2-3 in the course of which fasting and postprandial level (in 2 hours after breakfast) of glucose (GLC), and fasting insulin and C-peptide. Overall safety of metformin was confirmed -- throughout whole period of follow up with different variants of comparative analysis no cases of lactic acidosis were revealed. Practical lack of positive influence of metformin on glycemia at its initially not high level was accompanied with improvement of FC CHF, parameters of central hemodynamics, augmentation of functional capacities of patients, improvement of quality of life, lowering of number of decompensations of CHF and diminishment of degree of activation of SAS. It can be suggested that this dynamics is conditioned by the presence of cardioprotective properties in metformin what allows to recommend its application in patients with CHF and type 2 DM. PMID- 18429759 TI - [Peculiarities of cardiovascular syndrome in children and adolescents with obesity in dependence of level of insulin resistance]. AB - We conducted complex study of the state of cardiovascular system in 157 children aged 11-16 years with I-III degree obesity. Dazing on the results it is suggested to include in cardiovascular syndrome in children with obesity together with arterial hypertension syndrome of vegetative (autonomic) dysfunction, manifesting by disturbance of heart rate variability, endothelial dysfunction as well as systolic-diastolic dysfunction of the left ventricle. Degree of severity of impairment of cardiovascular system in children and adolescents with obesity depends on the level of insulin resistance. PMID- 18429760 TI - [Pulmonary hypertension and right ventricular failure. Part XIV. Differentiated therapy of primary (idiopathic) and associated forms of pulmonary arterial hypertension]. AB - In a series of articles the authors discuss literature data concerning epidemiology of pulmonary arterial hypertension (PAH), its modern classification, peculiarities of its pathogenesis and treatment in various diseases and conditions. The last 14-th communication contains consideration of issues of differentiated administration of modern pulmonary vasodilators to patients with PAH taking into account etiology and severity of the disease, availability of evidence based efficacy and safety data, method of application, and contraindications. In patients with PAH of II and III functional class (FC) endothelin receptor blocker bosentan is believed to be the drug of first choice. Bosentan can be administered orally. In patients with significant liver involvement phosphodiesterase inhibitor type 5 sildenafil should be used instead of bosentan for long term treatment of PAH. Sildenafil also can be taken orally. If bosentan is not sufficiently effective it can be combined with sildenafil; inhalations of prostanoid iloprost can be added to this combination when necessary. Is this tiple combination is not effective iloprost inhalations can be replaced by subcutaneous treprostinil or continuous intravenous infusion of epoprostenol. In patients with IV FC PAH therapy is started with subcutaneous administration of treprostinil or infusion of epoprostenol, while bosentan or/and sildenafil is added when necessary. PMID- 18429761 TI - [Controlled release metoprolol succinate in MERIT-HF. Analysis of patients subgroups]. AB - Results of the investigation of metoprolol CR/XL which was conducted in large randomized controlled study MERIT-HF in patients with chronic heart failure (CHF) are presented. In the whole trial this beta1-selective blocker lowered mortality of patients with CHF. Analysis of results in subgroups shows that metoprolol CR/XL was equally effective in middle aged and old patients, in men and women. PMID- 18429762 TI - [Peculiarities of cardiac involvement in patients with dystrophic myotonia]. AB - Dystrophic myotonia is a widespread hereditary disease. Pathological process touches development and functioning of various organs and tissues: smooth and skeletal muscular tissue, the heart, organ of vision, brain. Pathology of the heart serves as one of frequent manyfestations of dystrophic myotonia. Degenerative changes and fatty infiltration of myocardium not infrequently leads to dilated cardiomyopathy. Involvement of the heart in such patients is represented mainly by disturbance of cardiac rhythm and conduction. Course of the disease is progressive. PMID- 18429763 TI - [Alcoholic and dilated cardiomyopathy. Is sign of equality justified]. PMID- 18429764 TI - Antarctic terrestrial life--challenging the history of the frozen continent? AB - Antarctica is a continent locked in ice, with almost 99.7% of current terrain covered by permanent ice and snow, and clear evidence that, as recently as the Last Glacial Maximum (LGM), ice sheets were both thicker and much more extensive than they are now. Ice sheet modelling of both the LGM and estimated previous ice maxima across the continent give broad support to the concept that most if not all currently ice-free ground would have been overridden during previous glaciations. This has given rise to a widely held perception that all Mesozoic (pre-glacial) terrestrial life of Antarctica was wiped out by successive and deepening glacial events. The implicit conclusion of such destruction is that most, possibly all, contemporary terrestrial life has colonised the continent during subsequent periods of glacial retreat. However, several recently emerged and complementary strands of biological and geological research cannot be reconciled comfortably with the current reconstruction of Antarctic glacial history, and therefore provide a fundamental challenge to the existing paradigms. Here, we summarise and synthesise evidence across these lines of research. The emerging fundamental insights corroborate substantial elements of the contemporary Antarctic terrestrial biota being continuously isolated in situ on a multi-million year, even pre-Gondwana break-up timescale. This new and complex terrestrial Antarctic biogeography parallels recent work suggesting greater regionalisation and evolutionary isolation than previously suspected in the circum-Antarctic marine fauna. These findings both require the adoption of a new biological paradigm within Antarctica and challenge current understanding of Antarctic glacial history. This has major implications for our understanding of the key role of Antarctica in the Earth System. PMID- 18429765 TI - Physiological ecology of aquatic overwintering in ranid frogs. AB - In cold-temperate climates, overwintering aquatic ranid frogs must survive prolonged periods of low temperature, often accompanied by low levels of dissolved oxygen. They must do so with the energy stores acquired prior to the onset of winter. Overwintering mortality is a significant factor in their life history, occasionally reaching 100% due to freezing and/or anoxia. Many species of northern ranid frogs overwinter in the tadpole stage, which increases survival during hypoxic episodes relative to adults, as well as allowing for larger sizes at metamorphosis. At temperatures below 5 degrees C, submerged ranid frogs are capable of acquiring adequate oxygen via cutaneous gas exchange over a wide range of ambient oxygen partial pressures (PO(2)), and possess numerous physiological and behavioural mechanisms that allow them to maintain normal rates of oxygen uptake across the skin at a relatively low PO(2). At levels of oxygen near and below the critical PO(2) that allows for aerobic metabolism, frogs must adopt biochemical mechanisms that act to minimise oxygen utilisation and assist in maintaining an aerobic state to survive overwintering. These mechanisms include alterations in mitochondrial metabolism and affinity, changes in membrane permeability, alterations in water balance, and reduction in cellular electrochemical gradients, all of which lead to an overall reduction in whole animal metabolism. Winter energetic requirements are fueled by the energy stores in liver, muscle, and fat depots, which are likely to be sufficient when the water is cold and well oxygenated. However, under hypoxic conditions fat stores cannot be utilised efficiently and glycogen stores are used up rapidly due to recruitment of anaerobiosis. Since ranid frogs have minimal tolerance to anoxia, it is untenable to suggest that they spend a significant portion of the winter buried in anoxic mud, but instead utilise a suite of behavioural and physiological mechanisms geared to optimal survival in cold, hypoxic conditions. PMID- 18429766 TI - A new perspective on Darwin's Pangenesis. AB - In 1868 Charles Darwin proposed Pangenesis, a developmental theory of heredity. He suggested that all cells in an organism are capable of shedding minute particles he called gemmules, which are able to circulate throughout the body and finally congregate in the gonads. These particles are then transmitted to the next generation and are responsible for the transmission of characteristics from parent to offspring. If any cells of the parent undergo changes as a result of environmental change, they will consequently transmit modified gemmules to their offspring. Soon after Darwin's pangenetic theory was published, Francis Galton designed a series of blood transfusion experiments on differently pigmented rabbits to test its validity. He found no evidence in support of the existence of Darwin's gemmules and the concept of Pangenesis was largely abandoned. In this article, recent reports of successful induction of heritable changes by blood transfusion are reviewed. Detection of circulating nucleic acids and prions in plant sap and animal blood is considered as fresh evidence for the existence of gemmules. It is now apparent that a considerable revision of views on Darwin's Pangenesis must occur before a new comprehensive genetic theory can be achieved. PMID- 18429767 TI - An integrative view of sexual selection in Tribolium flour beetles. AB - Sexual selection is a major force driving the evolution of diverse reproductive traits. This evolutionary process is based on individual reproductive advantages that arise either through intrasexual competition or through intersexual choice and conflict. While classical studies of sexual selection focused mainly on differences in male mating success, more recent work has focused on the differences in paternity share that may arise through sperm competition or cryptic female choice whenever females mate with multiple males. Thus, an integrative view of sexual selection needs to encompass processes that occur not only before copulation (pre-mating), but also during copulation (peri-mating), as well as after copulation (post-mating), all of which can generate differences in reproductive success. By encompassing mechanisms of sexual selection across all of these sequential reproductive stages this review takes an integrative approach to sexual selection in Tribolium flour beetles (Coleoptera: Tenebrionidae), a particularly well-studied and economically important model organism. Tribolium flour beetles colonize patchily distributed grain stores, and juvenile and adult stages share the same food resources. Adults are highly promiscuous and female reproduction is distributed across an adult lifespan lasting approximately 1 year. While Tribolium males produce an aggregation pheromone that attracts both sexes, there appears to be little pre-mating discrimination among potential mates by either sex. However, recent work has revealed several peri-mating and post mating mechanisms that determine how offspring paternity is apportioned among a female's mates. During mating, Tribolium females reject spermatophore transfer and limit sperm numbers transferred by males with low phenotypic quality. Although there is some conflicting evidence, male copulatory leg-rubbing appears to be associated with overcoming female resistance to insemination and does not influence a male's subsequent paternity share. Evidence suggests that Tribolium beetles have several possible post-mating mechanisms that they may use to bias paternity. Male sperm precedence has been extensively studied in Tribolium spp. and the related Tenebrio molitor, and several factors influencing male paternity share among a female's progeny have been identified. These include oviposition time, inter-mating interval, male strain/genotype, the mating regimen of a male's mother, male starvation, and tapeworm infection. Females exert muscular control over sperm storage, although there is no evidence to date that females use this to differentiate among mates. Females could also influence offspring paternity by re-mating with additional males, and T. castaneum females more readily accept spermatophores when they are re-mating with more attractive males. Additional work is needed to examine the possible roles played by both male and female accessory gland products in determining male paternity share. Sexual selection during pre-mating episodes may be reinforced or counteracted by peri- and post copulatory selection, and antagonistic coevolution between the sexes may be played out across reproductive stages. In Tribolium, males' olfactory attractiveness is positively correlated with both insemination success and paternity share, suggesting consistent selection across different reproductive stages. Similar studies across sequential reproductive stages are needed in other taxa to provide a more integrative view of sexual selection. PMID- 18429770 TI - Pediatric teledermatology--a survey of current use. AB - Demand for pediatric dermatologic care far exceeds the supply of pediatric dermatologists. Teledermatology has been proposed as a solution to improve access to care, however, data regarding teledermatology use for pediatric patients are lacking. Surveys assessing use and experience with teledermatology were administered to pediatric dermatologists attending the 2006 Society for Pediatric Dermatology Annual Meeting and to programs offering teledermatology identified in the American Telemedicine Association's 2003 Teledermatology Survey. Over half of the 76 pediatric dermatologists responding to the survey had previous experience with teledermatology and nearly half currently use some form of teledermatology. For most, this represents nonreimbursed, store-and-forward consultations. Respondents noted that teledermatology allows more accurate triage of dermatology patients, decreases travel and outpatient clinic visits, and provides an avenue for ongoing support and education for primary care physicians. While difficulties exist, overcoming obstacles to the use of pediatric teledermatology and reimbursement for such services could improve access to expert pediatric dermatologic care. PMID- 18429769 TI - Cutaneous manifestations of cystic fibrosis. AB - Cystic fibrosis is an autosomal recessive disease reported in 1 in 2500 live births in Northern American and Northern European Caucasian populations. Classic disease findings include chronic bacterial infection of airways and sinuses, malabsorption of fat, infertility in men, and elevated concentrations of chloride in sweat. Less well-recognized findings associated with cystic fibrosis include cutaneous findings, which can be primary or secondary manifestations of the disease process. Patients demonstrate more atopic and drug hypersensitivity reactions than the general population, but have similar rates of urticaria compared with the general population. In atypical presentations of cystic fibrosis, the nutrient deficiency dermatitis of the disease may aid with diagnosis, and notably can be the presenting sign. Other dermatologic manifestations of cystic fibrosis include early aquagenic skin wrinkling and cutaneous vasculitis, which can be associated with arthralgias. Familiarity with the nutrient deficiency dermatitis of this entity may play a role in the timely diagnosis of the disease, and the other cutaneous findings add to our understanding of the protean nature of its manifestations. PMID- 18429771 TI - Discoid lupus erythematosus in children--a retrospective study of 34 patients. AB - Discoid lupus erythematosus is much less frequent and studied in children. We undertook a retrospective study of 34 children less than 16 years of age with this disease, seen over a period of 9 years. A female predominance of 2:1 was found. An association between discoid lupus erythematosus and systemic lupus erythematosus was observed in 23.5% of patients, a higher proportion compared to adult discoid lupus erythematosus. Disseminated lesions were much more frequent in patients with criteria for systemic lupus erythematosus (87.5% vs 34%), suggesting that it could be associated with a worse prognosis. Histologic findings were similar to those observed in adult discoid lupus erythematosus. PMID- 18429772 TI - Infantile hemangiomas: how common are they? A systematic review of the medical literature. AB - No published prospective studies have been published for several decades examining the incidence of hemangiomas. Older studies were performed before the delineation of "hemangiomas" from other vascular birthmarks was well-established. The objective of our study is to critically re-examine the literature reporting the incidence of infantile hemangiomas to determine if the true incidence is actually known. We performed both an electronic database search and hand search of the medical literature on the natural history of hemangiomas in full-term newborns and infants. A total of seven articles were found comprising two study populations: newborns <2 weeks of age and infants over the age of 2 weeks. All studies included samples sizes >500 patients including both hospital-based and primary care settings. Study designs ranged from retrospective chart reviews to cross-sectional cohort studies. Descriptive nomenclature was not uniform between studies, and all had methodologic limitations including problems of definition and study design. Studies estimating the true incidence of infantile hemangiomas are all many decades old and have significant methodologic issues limiting their ability to determine hemangioma incidence. Future studies in primary care settings using the currently accepted classification schema of vascular birthmarks may more accurately define the incidence and potential impact of this common vascular tumor of infancy. PMID- 18429773 TI - The effect of daily treatment with an olive oil/lanolin emollient on skin integrity in preterm infants: a randomized controlled trial. AB - To date, appropriate skin therapy for premature infants has not been clearly defined. Emollient creams are often used without solid evidence for a benefit to the neonate. The aim of the current study was to investigate the cutaneous effects of two different topical ointment therapies. Between October 2004 and November 2006 we prospectively enrolled 173 infants between 25 and 36 weeks of gestation admitted to a neonatal intensive care unit. Infants were randomly assigned to daily topical treatment with water-in-oil emollient cream (Bepanthen), olive oil cream (70% lanolin, 30% olive oil), or to a control group. Each neonate was continuously treated for a maximum of 4 weeks. Skin condition (skin score reflecting degree of dermatitis) in these groups was compared at weeks 1, 2, 3, and 4. Neonates treated with olive oil cream showed statistically less dermatitis than did neonates treated with emollient cream, and both had a better outcome than those in the control group (p < 0.001 in weeks 2-4). Treatment effects persisted throughout the study period and applied to infants of all gestational ages. This study demonstrates that topical skin therapy lowers the risk of dermatitis. Olive oil cream was superior to water-in-oil emollient cream. PMID- 18429774 TI - Oral cephalexin for acne vulgaris: clinical experience with 93 patients. AB - For patients who fail traditional acne therapy or experience side effects, other treatments are needed. Cephalexin has been noted to be efficacious in some instances. This study aimed to assess the therapeutic efficacy of cephalexin for acne. A retrospective chart review of 93 acne patients treated with cephalexin was performed. Data collection included patient demographics, other acne therapies, clinical response, and side effects. Of the 98 courses of cephalexin, 4% of patients cleared, 45% were much improved, 29% were somewhat improved, 16% experienced no change, and 6% worsened at first follow-up visit. The median treatment length was 6 months. Eighty-four percent of patients had previously used a systemic antibiotic(s) for acne. Seven percent experienced adverse effects. This is a small retrospective analysis without a comparison group and is limited by the number of patients and nonstandardized treatment lengths and charting procedures. Cephalexin is a promising alternative treatment for acne, with 78% of patients exhibiting at least some clinical improvement. PMID- 18429775 TI - Systemic methotrexate treatment in childhood psoriasis: further experience in 24 children from India. AB - Well-designed studies on systemic therapeutic modalities for severe psoriasis in children are rare. Children with severe disease are treated with the support of data extrapolated from that in adult, although management in them differs from adults in several important aspects. Like other systemic modalities, data regarding the use of methotrexate in the treatment of childhood psoriasis is meager. This study aims to analyze the efficacy and safety of methotrexate in severe or disabling childhood psoriasis. The records of all the patients <18 years of age treated with systemic methotrexate at the psoriasis clinic of our institute from January 1993 to December 2006 were retrieved. Information regarding demographic profile, disease characteristics, response to treatment, side effects, etc. was noted from predesigned clinic proforma. Indications of methotrexate use were baseline psoriasis area and severity index (PASI) >10, disease refractory to conventional therapies and disabling psoriasis even though the psoriasis area and severity index was <10. Clinical status of patients was assessed at weekly intervals for the first 2 weeks, fortnightly during next month and then monthly. Response to therapy was graded as good (50-75% decrease in PASI) and excellent (>75% decrease in PASI). Laboratory investigations to detect methotrexate induced toxicity were performed at regular intervals. Of the 29 patients treated with methotrexate, 24 were eligible for the final data analysis. Indication for the institution of methotrexate therapy was severe disease, viz., extensive recalcitrant plaque type psoriasis in 17 patients, erythroderma and generalized pustular psoriasis of von-Zumbusch type in three patients each and severe disabling palmo-plantar involvement along with chronic plaque lesions in one patient. Response to therapy was excellent (>75% decrease in PASI) in all but two patients. The mean time to control the disease, i.e., 50% reduction in PASI was 5.1 weeks. Mean total cumulative dose of methotrexate in the first episode was 215 mg. The duration of remission could be calculated in nine patients only, varying from 1.5 months to 3 years. Side effects were mild, observed in nine children, which included nausea, vomiting, and loss of appetite. Methotrexate is an effective, cheap, easily available, and reasonably safe drug to be used in severe childhood psoriasis under an expert supervision and laboratory monitoring. PMID- 18429776 TI - One-year experience with candida antigen immunotherapy for warts and molluscum. AB - Molluscum contagiosum and warts are two very common viral-induced diseases of childhood. We report our 1-year experience with intralesional candida antigen therapy for both warts and molluscum contagiosum. A retrospective chart review was performed and a total of 217 patients were identified. Follow-up was obtained either through clinic visits or telephone interviews for 25/47 molluscum patients and 55/170 wart patients. Of the molluscum patients with follow-up, 14/25 (56%) had complete resolution, 7/25 (28%) had partial clearing, and 4/25 (16%) had no improvement. For the wart patients with follow-up, 48/55 (87%) had complete resolution, 3/55 (6%) had partial clearing, and 4/55 (7%) had no improvement. It is important to note that our clearance rate may be overestimated, as many patients were also treated concurrently with liquid nitrogen or other therapies. All wart and molluscum patients experienced some discomfort at the time of injection, but no serious side effects were reported. Our experience suggests intralesional candida antigen may represent a treatment option for warts and molluscum contagiosum that is safe and effective. PMID- 18429777 TI - Cutaneous and paravertebral infantile hemangioma: report of two cases. AB - We report on two cases of segmental cutaneous infantile hemangioma associated with mediastinal hemangioma and paravertebral extension. Mediastinal involvement is rare in hemangioma and paravertebral and intraspinal extension of hemangioma have seldom reported and may have been overlooked in the past. PMID- 18429778 TI - Treatment of granuloma annulare with the 595 nm pulsed dye laser in a pediatric patient. PMID- 18429779 TI - Phacomatosis pigmentovascularis type Va in a 3-month old. AB - Phacomatosis pigmentovascularis is a rare genodermatosis characterized by a combination of melanocytic nevi and vascular malformations. A new type of phacomatosis pigmentovascularis was recently described which included cutis marmorata telangiectatica congenita and aberrant Mongolian spots and was named type V. We report a 3-month-old girl with diagnosis of phacomatosis pigmentovascularis type V. PMID- 18429780 TI - Vesicular erythema multiforme-like reaction to para-phenylenediamine in a henna tattoo. AB - Allergic contact dermatitis reaction to topical "black henna" tattoo is usually described secondary to the organic dye para-phenylenediamine, a derivative of analine. Allergic contact dermatitis reactions to para-phenylenediamine are well recognized and most commonly involve an eczematous reaction that may become generalized and an acute angio-edema. Only four previous instances have been reported of an erythema multiforme-like reaction to para-phenylenediamine and its derivatives, including only one mild reaction to a tattoo. A vesicular erythema multiforme-like reaction has not been reported. An erythema multiforme-like reaction to contact allergens is usually caused by potent allergens including plant quinolones in Compositae and sesquiterpene lactones in exotic woods, and it is also reported to topical drugs, epoxy resin, metals (particularly nickel), and various chemicals. A generalized vesicular erythema multiforme-like reaction is unusual, and rarely reported. We describe a 6-year-old boy who developed a localized, eczematous and severe generalized vesicular erythema multiforme-like contact allergy to para-phenylenediamine secondary to a henna tattoo. As henna tattoos are becoming increasingly popular, one should be aware of the possibility of such a reaction. This presentation also highlights the call to ban the use of para-phenylenediamine and its derivatives in dyes. PMID- 18429781 TI - Unilateral mosaic cutaneous vascular lesions, enchondroma, multiple soft tissue chondromas and congenital fibrosarcoma--a variant of Maffucci syndrome? AB - The coexistence of enchondromas and vascular lesions characterizes the principal feature of a rare congenital condition known as Maffucci syndrome. We present a 20-year-old male with a mosaic distribution of vascular malformations and atrophic overlying dermis and subcutis. Enchondroma and multiple periosteal chondromas of the ipsilateral limb led to the working diagnosis of Maffucci syndrome. Of interest, this patient also has a history of congenital fibrosarcoma with concomitant thrombocytopenia. To our knowledge this is the first report of this constellation of findings, which may represent a form of mesenchymal mosaicism analogous to the Blaschkoid distribution of other genodermatoses. PMID- 18429782 TI - Dystrophic epidermolysis bullosa with one dominant and one recessive mutation of the COL7A1 gene in a child with deafness. AB - Dystrophic epidermolysis bullosa can be inherited in autosomal dominant and recessive forms, the former usually expressed as a milder phenotype, although mild forms of recessive dystrophic epidermolysis bullosa can occur. We present a patient who was found to be a compound heterozygote, inheriting a dominant mutation from his father and a recessive mutation from his mother, resulting in a clinically severe case of dystrophic epidermolysis bullosa. Mutations in the gene for collagen VII (COL7A1) have been documented in both types of dystrophic epidermolysis bullosa. Our patient has also been diagnosed with bilateral auditory neuropathy, a disorder coincidentally also mapped to a nearby gene on chromosome 3p21 (the transmembrane inner ear expressed gene, TMIE). PMID- 18429783 TI - Addison's disease, diffuse skin, and mucosal hyperpigmenation with subtle "flu like" symptoms--a report of two cases. AB - Addison's disease, or chronic adrenocortical insufficiency, is the overproduction of adrenocorticotropic hormone by the pituitary gland as a compensatory mechanism for decreased cortisol production by the adrenal glands. Classically, patients affected with Addison's disease develop weakness, anorexia, electrolyte imbalances: decreased sodium and chloride with increased serum potassium resulting in hypotension, and hyperpigmentation of the skin and mucous membranes. Herein this case report, we focus on the subtle findings of diffuse hyperpigmentation and intermittent but repetitive "flu-like" symptoms in two patients to correctly identify the diagnosis of Addison's disease effectively and efficiently. PMID- 18429784 TI - Body piercing complicated by atypical mycobacterial infections. AB - Body piercing is a growing trend, especially in young people, but the literature on complications of piercing consists mostly of case reports involving ear piercing. Previous reported complications of piercing include contact dermatitis, keloids, traumatic tearing, viral transmission, and bacterial infections. We report two patients who presented with atypical mycobacterial infections of body piercing sites. It is important to recognize the association of piercing and mycobacterial infections so that tissue can be obtained for histopathologic examination and appropriate culture. PMID- 18429785 TI - Palmoplantar keratoderma and skin grafting: postsurgical long-term follow-up of two cases with Olmsted syndrome. AB - Olmsted syndrome is a rare congenital mutilating palmoplantar keratoderma associated with periorificial keratotic plaques. Treatment options include topical keratolytics, systemic retinoids, and debulking procedures. Full thickness excision of hyperkeratotic plaques followed by skin grafting has been reported in the medical literature, although long-term results have not been evaluated. We present two cases of Olmsted syndrome with severe palmoplantar keratoderma treated with excision and skin grafting, along with long-term clinical results 11 years (patient 1) and 6 years (patient 2) following the initial surgery. PMID- 18429786 TI - Zoonotic deep cutaneous filariasis--three pediatric cases from Quebec, Canada. AB - Three rare cases of pediatric Quebec-based zoonotic filarial nematode deep skin infections were reviewed. These rare cases were processed at our pediatric hospital within the last 6-year period. Patient age, travel information, lesional characteristics, systemic findings, serology, histopathology, treatment, and follow-up were gathered from the submitting specimen and the treating physicians. Species identification was performed by the Parasitic Disease Branch, Division of Infectious and Tropical Diseases Pathology, AFIP, Washington, DC. PMID- 18429787 TI - Superimposed Mongolian spots. AB - We report four Chinese infants with a darker pigmented Mongolian spot superimposed on another Mongolian spot. The presence of a superimposed Mongolian spot has not been previously reported. Mongolian spots can look similar to a bruise and awareness of these lesions is important to avoid unjustified consideration of injury or child abuse. PMID- 18429788 TI - Generalized congenital smooth muscle hamartoma presenting with hypertrichosis, excess skin folds, and follicular dimpling. AB - We describe an interesting patient with a diffuse smooth muscle hamartoma who presented with hypertrichosis, increased skin folds, and follicular dimpling. While smooth muscle hamartoma classically presents as a small hairy, skin-colored to hyperpigmented patch or plaque, rare presentations with diffuse involvement or follicular papules have been described. We present our patient and review the literature to highlight the diverse and under recognized manifestations of this benign neoplasm. PMID- 18429789 TI - Sclerodermatous chronic graft-versus-host disease--a report of four pediatric cases. AB - The sclerodermatous variant of chronic graft-versus-host disease postallogeneic bone marrow transplantation is rare. We present four pediatric cases of sclerodermatous variant of chronic graft-versus-host disease describing their clinical appearance, management, and outcomes. We compare the pharmacologic and supportive therapies administered to these patients with the management suggested in the current literature. Several key findings were noted. There was a significantly higher mortality rate observed in this series compared with previous reports, with three of the four patients dying ultimately as a result of sclerodermatous variant of chronic graft-versus-host disease. The development of widespread ulceration, in two of the four patients, appeared to be associated with an overall deterioration in the clinical condition. In two patients high dose thalidomide at 12 mg/kg/day seemed to halt the progression of cutaneous disease. Optimal care of sclerodermatous variant of chronic graft-versus-host disease patients required a multidisciplinary team. A lack of community services observed in this case series led to the need for unnecessarily prolonged inpatient admissions. PMID- 18429790 TI - Two cases of spontaneous stellate scars. AB - Scar formation is a physiologic reaction to dermal injury. We describe a 11-year old girl and her 15-year-old cousin, who both presented with stellate scarlike lesions primarily located on the hands. Histopathologic examination confirmed scar tissue formation. There was no history of initiating trauma or other possible triggering factors. To our knowledge, this is the first reported case of spontaneous stellate scars in two young, second-degree family members, the exact pathogenesis of which remains to be elucidated. PMID- 18429791 TI - Nail changes in Langerhans cell histiocytosis: a possible marker of multisystem disease. AB - We describe a child with a 6-month history of onycholysis, subungual hyperkeratosis, and hemorrhages in most of her fingernails and toenails. Nail involvement preceded the identification of osteolytic lesions at the mastoid on a cranial computed tomography scan, which was performed because of repeated episodes of acute otitis media. Some weeks later, a small number of erythematous papules developed over the trunk and face. The diagnosis of Langerhans cell histiocytosis was made by histopathologic examination of bone, skin, and bed and matrix nail biopsies. Response to treatment with vinblastine and prednisone was excellent. Nail changes in Langerhans cell histiocytosis are extremely uncommon, particularly as the presenting manifestation of the disease. The role of nail involvement as an unfavorable prognostic sign is unclear. PMID- 18429792 TI - Periorbital, facial, and scalp plaques since birth. PMID- 18429794 TI - Two cases of asymptomatic exophytic papules on head. PMID- 18429793 TI - An unusual dermatitis with annular lesions. PMID- 18429795 TI - Neutrophilic figurate erythema of infancy. AB - We report here a new case of neutrophilic figurate erythema of infancy in a 1 year and 9-month-old white boy. Neutrophilic figurate erythema of infancy is a rare inflammatory dermatosis which is part of the figurate inflammatory dermatoses of infancy and is considered a variant of annular erythema of infancy. The disease is clinically characterized by annular erythematous lesions, sometimes with a polycyclic configuration, and histologically by a dermal neutrophilic infiltration with leukocytoclasia. Differential diagnosis mainly includes clear-cut severe diseases with a well-known etiology, such as neonatal lupus erythematosus and its variant erythema gyratum atrophicans transiens neonatale, erythema chronicum migrans and erythema marginatum rheumaticum, and diseases of unknown origin and with less clear limits, such as erythema annulare centrifugum and its variant familial annular erythema. Anamnesis, laboratory findings, clinical features, and histology allow the correct diagnosis and therefore, having excluded severe diseases, parents should be reassured as neutrophilic figurate erythema of infancy, as the annular erythema of infancy, is a benign disease, in most cases unassociated with other conditions and usually self-limiting over a few months, even though a chronic course may rarely occur. Our case was characterized by a chronic persistent course and by a complete resolution of the lesions only during febrile episodes. PMID- 18429796 TI - Penile granuloma annulare of an adolescent male--case report and review of the literature. PMID- 18429797 TI - First case report of topical tacalcitol for vitiligo repigmentation. AB - We present a 11-year-old girl with an 8-month history of vitiligo vulgaris on her postcervical, periocular, and periauricular regions. The region topically treated with 0.0002% tacalcitol ointment (vitamin D3 ointment) and sunlight exposure healed completely, while the region treated with the ointment alone did not improve. A combination therapy with tacalcitol ointment and sunlight exposure instead of UV irradiation can be recommended as an alternative therapy for vitiligo vulgaris because the treatment is safe and increases compliance in children. PMID- 18429798 TI - A rare presentation of squamous cell carcinoma in a patient with PIBIDS-type trichothiodystrophy. AB - The clinical presentation of trichothiodystrophy type F includes photosensitivity, ichthyosis, brittle hair, intellectual impairment, decreased fertility, and short stature, often referred to as the PIBIDS syndrome. While many of these patients demonstrate features also found in xeroderma pigmentosum patients, including similar nucleotide excision repair gene defects and photosensitivity, PIBIDS patients rarely demonstrate cutaneous malignancies. This case report demonstrates the rare presentation of squamous cell carcinoma developing in a PIBIDS patient. PMID- 18429799 TI - Clinical and histopathologic study of pilomatricoma in Iran between 1992 and 2005. AB - Pilomatricoma (calcifying epithelioma of Malherb) is a tumor of hair follicle matrix cells and is almost always benign. In this study we reviewed the clinical and histopathological features of 45 cases of pilomatricoma during 13 years (1992 2005). Although few malignant transformations have been reported in other studies, no evidence of malignancy or aggressiveness is observed in this study. PMID- 18429800 TI - Suppressive effects of topical mometasone furoate and tacrolimus on skin prick testing in children. AB - Skin prick tests were performed in 12 children with atopic eczema before and after 2 weeks of treatment with topical mometasone furoate and tacrolimus. Both treatments significantly suppressed the allergen wheal size. Mometasone furoate reduced the histamine wheal size as well. Skin prick testing in children treated with topical glucocorticoids or tacrolimus is associated with a risk of false negative test results. PMID- 18429801 TI - Dermoscopic features in Bean (blue rubber bleb nevus) syndrome. AB - Dermoscopy has become an important tool for evaluating skin lesions in children. We report the dermoscopic findings in a case of Bean syndrome and emphasize the utility of this technique in vascular lesions. PMID- 18429803 TI - Temporary localized hypertrichosis after henna pseudotattoo. AB - We report the case of a 5-year-old boy who had a temporary dragon-shaped henna pseudotattoo, reinforced 4 days later. Two weeks later, as the pseudotattoo began to disappear, hypertrichosis developed in the area corresponding to the previous psuedotattoo. Skin biopsy showed an increase in vellus hair follicles, with slight peripheral fibrosis. After 4 months, the hypertrichosis resolved spontaneously. PMID- 18429802 TI - Lentigines within nevus depigmentosus: a rare collateral effect of UVB therapy? AB - Nevus depigmentosus is an uncommon hypopigmented macule or patch that is congenital and stable in its relative size and distribution throughout life; it occurs sporadically and may be localized, segmental or, less often, systematized. We report the case of a 17-year-old girl with a segmental achromic nevus of the left leg and a patch on the lower back with late age of onset who developed lentigines after prolonged intense ultraviolet B exposure as a consequence of an incorrect diagnosis of segmental vitiligo. PMID- 18429804 TI - Successful treatment of recalcitrant warts with topical squaric acid in immunosuppressed child. AB - Although spontaneous resolution of verrucous vulgaris is the norm, many treatment modalities are available including local destruction, chemotherapy, immunotherapy, and sensitizing agents. The choice of treatment will vary with patient's age and desire for treatment, prior treatments, potential side effects, location of lesions, and patient's immune status. Treatment of verruca vulgaris in immunosuppressed patients presents therapeutic challenges as lesions in this population are often refractory to conventional modalities. Here we present a case of successful treatment of recalcitrant warts with topical squaric acid in an immunosuppressed child. PMID- 18429805 TI - Pediatric skin disorders encountered in a dermatology outpatient clinic in Turkey. AB - This study documents the spectrum and frequency of skin disorders in a total of 2370 children encountered in a dermatology outpatient clinic in Turkey. Infectious skin diseases were most frequently observed (27.6%) followed by, eczemas (17.9%), acne (14,5%), papulosquamous diseases (6.9%), hair diseases (4.1%), pigmentation disorders (3.8%), generalized pruritus (2.8%), urticaria (2.7%), and insect bite (2.3%). Epidemiologic data are useful in planning of the health care and taking preventive measures to decrease the prevalence of skin disorders in children. PMID- 18429806 TI - Beau's lines and multiple periungueal pyogenic granulomas after long stay in an intensive care unit. AB - A child developed multiple Beau's lines and periungueal pyogenic granulomas after admission to the intensive care unit. Immobilization, hypoxia, and drugs might have acted as potential causative factors. PMID- 18429807 TI - Bullous congenital ichthyosiform erythroderma associated with hypocalcemic vitamin D-resistant rickets. AB - There are several reports of an association between ichthyosis and rickets based mainly on a causative relationship through the impaired ability of ichthyotic skin to synthesize vitamin D. Hypocalcemic vitamin D-resistant rickets represents a specific type of rickets that is attributed to vitamin D receptor defect rather than to vitamin D deficiency. Nevertheless, an extremely rare association between bullous congenital ichthyosiform erythroderma and hypocalcemic vitamin D resistant rickets is presented. PMID- 18429808 TI - Commentary--Complications of body piercing. PMID- 18429809 TI - Iatrogenic neonatal burns--more causes. PMID- 18429810 TI - Mongolian spots in Sjogren-Larsson syndrome. PMID- 18429811 TI - Onychomatricome: first description in a child. PMID- 18429812 TI - Netherton syndrome and trichorrhexis invaginata--a novel diagnostic approach. PMID- 18429813 TI - Familial mucinous nevus. PMID- 18429814 TI - ERK5 is involved in TCR-induced apoptosis through the modification of Nur77. AB - Nur77 is a nuclear orphan steroid receptor that has been implicated in negative selection when immature T cells are strongly activated through interaction with self peptide-MHC complexes. The expression of Nur77 in thymocytes and T cell lines leads to apoptosis in a manner dependent on its transcriptional activity. It is well established that Nur77 function is negatively regulated by post translational modification. Here we demonstrate that the MAPK-induced phosphorylation of Nur77 during T cell activation plays a critical role in the induction of apoptosis. Upon T cell receptor (TCR) stimulation, ERK5 (also known as big MAP kinase 1, BMK1), a member of the MAPK family, phosphorylates Nur77, leading to its transcriptional activation. In contrast, the activation of the ERK2 signaling pathway failed to activate Nur77 although ERK2 is also able to phosphorylate Nur77. Furthermore, the blockade of ERK5 signaling pathway suppressed TCR-induced cell death. These results indicate that ERK5 regulates Nur77 function through its phosphorylation. PMID- 18429815 TI - Lyonization pattern of normal human nails. AB - To examine the X-inactivation patterns of normal human nails, we performed the human androgen receptor gene assay of DNA samples extracted separately from each finger and toe nail plates of nine female volunteers. The X-inactivation pattern of each nail was unique and constant for at least 2 years. The frequency of nails with one of the two X-chromosomes exclusively inactivated was 25.9%. In the nails composed of two types of cells with either one X-chromosome inactivated, the two cell types were distributed in patchy mosaics. These findings suggest that the composition of precursor cells of each nail is maintained at each site at least through several cycles of regeneration time, and that the nail plate has a longitudinal band pattern, each band consisting of cells with only one of the two X-chromosomes inactivated. Using the frequency of nails with one of two X chromosomes exclusively inactivated, we estimated the number of progenitor cells that gave rise to the nail plate during development to be about 3, under the assumption that the process follows the binominal distribution model. A strong correlation observed among the big, index and little fingers, and among the corresponding toes suggests an interesting interpretation concerning their morphogenetic process. PMID- 18429816 TI - HMRF1L is a human mitochondrial translation release factor involved in the decoding of the termination codons UAA and UAG. AB - While all essential mammalian mitochondrial factors involved in the initiation and elongation phases of translation have been cloned and well characterized, little is known about the factors involved in the termination process. In the present work, we report the functional analysis of human mitochondrial translation release factors (RF). Here, we show that HMRF1, which had been previously denoted as a human mitochondrial RF, was inactive in in vitro translation system, although it is a mitochondrial protein. Instead, we identified another human mitochondrial RF candidate, HMRF1L, and demonstrated that HMRF1L is indeed a mitochondrial protein that functions specifically as an RF for the decoding of mitochondrial UAA and UAG termination codons in vitro. The identification of the functional mitochondrial RF brings us much closer to a detailed understanding of the translational termination process in mammalian mitochondria as well as to the unraveling of the molecular mechanism of diseases caused by the dys-regulation of translational termination in human mitochondria. PMID- 18429817 TI - Bood POZ containing gene type 2 is a human counterpart of yeast Btb3p and promotes the degradation of terminal deoxynucleotidyltransferase. AB - Bood POZ containing gene type 2 (BPOZ-2) is involved in the growth suppressive effect of the phosphatase and tensin homologue (PTEN). We showed that BPOZ-2 is a human counterpart of yeast Btb3p, which is a putative adaptor for Pcu3p-based ubiquitin ligase. BPOZ-2 bound to E3 ligase CUL3 in vitro and in vivo. BPOZ-2 itself was ubiquitinated through the CUL3-based E3 ligase mainly within the nucleus and degraded by the 26S proteasome. Although BPOZ-2 was mainly expressed within the cytoplasm, it accumulated within the nucleus in the presence of the specific 26S proteasome inhibitor MG132. BPOZ-2 may be recruited to the nucleus from the cytoplasm. Terminal deoxynucleotidyltransferase (TdT) was detected as a BPOZ-2-binding protein using a yeast two-hybrid system by screening a human thymus cDNA library. TdT, BPOZ-2, and CUL3 formed a ternary complex in vivo. TdT was ubiquitinated only within the nucleus and degraded by the 26S proteasome. The ubiqutination or degradation of TdT was markedly promoted by co-expression of BPOZ-2 and CUL3 or BPOZ-2 in 293T cells, respectively. PMID- 18429818 TI - Spontaneous mutagenesis associated with nucleotide excision repair in Escherichia coli. AB - The vast majority of spontaneous mutations occurring in Escherichia coli are thought to be derived from spontaneous DNA lesions, which include oxidative base damage. Systems for removing intrinsic mutagens and repairing DNA lesions contribute to the suppression of spontaneous mutations. Nucleotide excision repair (NER) is a general DNA repair system that eliminates various kinds of lesions from DNA. We therefore predicted that NER might be involved in suppression of spontaneous mutations, and analyzed base substitutions occurring spontaneously within the rpoB gene in NER-proficient (wild-type), -deficient and overproducing E. coli strains. Surprisingly, the mutation frequency was lower in NER-deficient strains, and higher in NER-overproducing strains, than in the NER proficient strain. These results suggest, paradoxically, that NER contributes to the generation of spontaneous mutation rather than to its suppression under normal growth conditions, and that transcription-coupled repair also participates in this process. Using E. coli strains that carried an editing exonuclease deficient polA mutation, we further obtained data suggesting that unnecessary NER might account for these findings, so that errors introduced during repair DNA synthesis by DNA polymerase I would result in unwanted base substitutions. The repair system itself may thus be an important generator of spontaneous mutation. PMID- 18429819 TI - Inhibition of filament formation of human Rad51 protein by a small peptide derived from the BRC-motif of the BRCA2 protein. AB - Human Rad51 is a key element of recombinational DNA repair and is related to the resistance of cancer cells to chemo- and radiotherapies. The protein is thus a potential target of anti-cancer treatment. The crystallographic analysis shows that the BRC-motif of the BRCA2 tumor suppressor is in contact with the subunit subunit interface of Rad51 and could thus prevent filament formation of Rad51. However, biochemical analysis indicates that a BRC-motif peptide of 69 amino acids preferentially binds to the N-terminal part of Rad51. We show experimentally that a short peptide of 28 amino acids derived from the BRC4 motif binds to the subunit-subunit interface and dissociates its filament, both in the presence and absence of DNA, certainly by binding to dissociated monomers. The inhibition is efficient and specific for Rad51: the peptide does not even interact with Rad51 homologs or prevent their interaction with DNA. Neither the N terminal nor the C-terminal half of the peptide interacts with human Rad51, indicating that both parts are involved in the interaction, as expected from the crystal structure. These results suggest the possibility of developing inhibitors of human Rad51 based on this peptide. PMID- 18429820 TI - BREK/LMTK2 is a myosin VI-binding protein involved in endosomal membrane trafficking. AB - Myosin VI is involved in a wide range of endocytic and exocytic membrane trafficking pathways; clathrin-mediated endocytosis, intracellular transport of clathrin-coated and -uncoated vesicles, AP-1B-dependent basolateral sorting in polarized epithelial cells and secretion from the Golgi complex to the cell surface. In this study, using a yeast two-hybrid screen, we identified brain enriched kinase/lemur tyrosine kinase 2 (BREK/LMTK2), a transmembrane serine/threonine kinase with previously unknown cellular functions, as a myosin VI-interacting protein. Several binding experiments confirmed the interaction of myosin VI with BREK in vivo and in vitro. Immunocytochemical analyses revealed that BREK localizes to cytoplasmic membrane vesicles and to perinuclear recycling endosomes. Notably, cells in which BREK was depleted by siRNA were still able to internalize transferrin molecules and to transport them to early endosomes, but were unable to transport them to perinuclear recycling endosomes. Our results show that BREK is critical for the transition of endocytosed membrane vesicles from early endosomes to recycling endosomes and also suggest an involvement of myosin VI in this pathway. PMID- 18429821 TI - Scheduled exposures to a novel environment with a running-wheel differentially accelerate re-entrainment of mice peripheral clocks to new light-dark cycles. AB - Effects of scheduled exposures to novel environment with a running-wheel were examined on re-entrainment to 8 h shifted light-dark (LD) cycles of mouse circadian rhythms in locomotor activity and clock gene, Per1, expression in the suprachiasmatic nucleus (SCN) and peripheral tissues. Per1 expression was monitored by a bioluminescence reporter introduced into mice. The animals were exposed to the novel environment for 3 h from the shifted dark onset for four cycles and released into constant darkness. In the phase-advance shift, the circadian rhythm in locomotor activity fully re-entrained in the exposed group, whereas it was in transients in the control. On the other hand, the circadian rhythm of Per1 expression in the SCN almost completely re-entrained in both the control and exposed groups. In the skeletal muscle and lung, the circadian rhythm fully re-entrained in the exposed group, whereas the rhythms in the control did not. In the phase-delay shift, the circadian rhythms in locomotor activity and Per1 expression almost completely re-entrained in both groups. These findings indicate that the scheduled exposures to novel environment with a running-wheel differentially accelerate the re-entrainment of the mouse peripheral clocks to 8 h phase-advanced LD cycles. PMID- 18429822 TI - Negative regulation of constitutive NF-kappaB and JNK signaling by PKN1-mediated phosphorylation of TRAF1. AB - Inhibitor of NF-kappaB (IkappaB) kinase (IKK) and c-Jun NH(2)-terminal kinase (JNK) are stress inducible kinases that critically regulate numerous physiological and pathological processes. Transient activation of the downstream transcription factors NF-kappaB and AP-1, allows for stress inducible, inflammatory and innate immune gene expression programs. However, elevated chronic activity is associated with cancer and chronic inflammatory disease. Despite its relevance to human health, little is known about the molecular mechanisms that control constitutive activity of IKK and JNK. Here, we demonstrate that the serine/threonine kinase PKN1 plays a critical role in regulating constitutive IKK/JNK activity in unstimulated cells and report on the molecular mechanism. We identify TRAF1 as a substrate of PKN1 kinase activity in vitro and in vivo, and show that this phosphorylation event is required for attenuating downstream kinase activities. Furthermore, this silencing was dependent on TNFR2. Mutagenesis of the phospho-acceptor residue in TRAF1 abrogated PKN1-dependent recruitment to TNFR2. Our results suggest a model by which the stoichiometric ratio of TRAF1 and TRAF2 heteromeric complexes associated with TNFR2 control the tonic activity of JNK and IKK. TRAF1 phosphorylation by the ubiquitously expressed kinase PKN1 thereby plays a critical role in the negative regulation of tonic activity of the two central inflammatory signaling pathways. PMID- 18429823 TI - Roles of neural stem progenitor cells in cytomegalovirus infection of the brain in mouse models. AB - Cytomegalovirus (CMV) is the most significant infectious cause of brain disorders in humans. Although the brain is the principal target organ for CMV infection in infants with congenital infection and in immunocompromised patients, little has been known about cellular events in pathogenesis of the brain disorders. Mouse models have been developed by the authors for studying the cell tropism, infectious dynamics of CMV infection and the effects of CMV infection on proliferation, regeneration and differentiation of neural cells. It has been shown, using brain slice cultures and neurospheres, that neural stem progenitor (NSP) cells are the most susceptible to CMV infection in developing brains. The NSP cells are also susceptible to CMV infection in adult and aged brains. The susceptibility can be enhanced by stimulation of neurogenesis. It was shown that latent murine CMV infection occurs in NSP cells by demonstrating the reactivation in brain slice culture or neurospheres. It is hypothesized that CMV brain disorder such as microcephaly is caused by disturbance of cellular events in the ventricular regions, including proliferation and differentiation of the neural stem cells, whereas neurons are also targets in persistent CMV infection, presumably resulting in functional disorders such as mental retardation. PMID- 18429824 TI - Pathological evaluation of uterine leiomyomas treated with gonadotropin-releasing hormone agonist (GnRH-a) therapy: role of mast cells and a possible mechanism of GnRH-a resistance in leiomyomas. AB - Gonadotropin-releasing hormone agonist (GnRH-a) therapy is frequently applied to reduce the volume of uterine leiomyomas (UL). In addition, the possible relationship between mast cells (MC) within UL and the development of UL has been suggested, but the role of MC in UL remains to be determined. UL with or without GnRH-a therapy in 121 premenopausal patients were reviewed. The number of MC was evaluated between the two groups, immunohistochemistry was done for insulin-like growth factor-I (IGF-I), and the association between the IGF-I immunoreactivity in UL and the GnRH-a therapy was analyzed. The number of MC significantly increased in UL in GnRH-a therapy, while IGF-I immunoreactivity was significantly reduced in smooth muscle cells of these UL. Furthermore, IGF-I immunoreactivity in MC was inversely correlated with the size reduction rate of UL in GnRH-a therapy. Although GnRH-a therapy is considered to reduce the size of UL transiently, the regression of UL was in part hampered by the increased IGF-I secretion from the increased MC after GnRH-a therapy. Therefore, the more the IGF I secretion from MC in UL increases, the less effective the GnRH-a therapy is on the size reduction of UL. Thus, the present study may provide an explanation of the possible mechanism of GnRH-a resistance in UL. PMID- 18429825 TI - Dinucleotide repeat polymorphisms of RAD51, BRCA1, BRCA2 gene regions in breast cancer. AB - Recent studies suggest that genetic polymorphisms of the DNA repair genes have been implicated in breast cancer risk. BRCA1 and BRCA2, two breast cancer susceptibility genes, are essential to maintain chromosomal integrity. This is mediated via regulation of RAD51 during homologous recombination. Dinucleotide polymorphism repeats in the 15q14-21, 17q21 and 13q12-13 regions, where the RAD51, BRCA1 and BRCA2 genes are located, respectively, have been evaluated. The polymorphism was determined using the following microsatellite markers: D15S118, D15S214, D15S1006, D17S855, D17S1323, D13S260 and D13S290. Genotypes containing the (CA)(17) or (CA)(19) alleles in the RAD51 region were found to be associated with a decreased breast cancer risk. Genotype containing the (CA)(17) allele in the 13q12-13 region was found to be associated with an increased breast cancer risk. The results indicate that dinucleotide CA repeat polymorphism at RAD51 and BRCA2 gene regions might be associated with genetic susceptibility to breast cancer. PMID- 18429826 TI - Lymphatic invasion according to D2-40 immunostaining is a strong predictor of nodal metastasis in superficial squamous cell carcinoma of the esophagus: algorithm for risk of nodal metastasis based on lymphatic invasion. AB - In squamous cell carcinoma (SCC) of the esophagus, D2-40 immunostaining has recently been used to detect lymphatic invasion, but invasion detected using D2 40 immunostaining for a predictor of nodal metastasis was controversial. Therefore, the usefulness of detecting lymphatic invasion by D2-40 immunostaining as a predictor of nodal metastasis was examined in superficial (mucosal and submucosal) SCC of the esophagus. A total of 115 superficial SCC of the esophagus were examined on immunohistochemistry using D2-40. It was found that lymphatic invasion demonstrated on D2-40 immunostaining was mainly detected in the lamina propria mucosa. Lymphatic invasion was found in 37 cases and the invasion detected in the entire tumor tissue was statistically correlated with nodal metastasis. Based on the lymphatic invasion according to D2-40 immunostaining, an algorithm was devised for the risk (low, intermediate and high) of nodal metastases in superficial SCC in the esophagus. In conclusion, the detection of lymphatic invasion on D2-40 immunostaining in tumor tissue is a strong predictor for nodal metastasis in superficial SCC of the esophagus. Lymphatic invasion was found mainly in the lamia propria mucosa, thus the devised algorithm is useful for determining the optimal treatment strategy after endoscopic mucosal resection for esophageal SCC. PMID- 18429827 TI - CD109 expression in basal-like breast carcinoma. AB - Breast cancer can be classified into several subtypes based on gene expression profiling. Basal-like breast carcinoma (BLC) has a triple negative phenotype, that is, the subtype lacks the estrogen receptor (ER), progesterone receptor (PgR) and human epidermal growth factor receptor 2 (HER2). It has been recently reported that CD109, a glycosylphosphatidylinositol (GPI)-anchored cell surface protein, is a new breast myoepithelial marker. In the present study CD109 expression was investigated in invasive ductal carcinomas (IDC) of the breast on immunohistochemistry. Eighty-eight formalin-fixed, paraffin-embedded breast carcinoma sections were immunostained with anti-CD109, anti-cytokeratin 5/6 (CK5/6), anti-calponin, anti-vimentin and anti-p63 antibodies. CD109 expression was detected in 18 of 30 basal-like breast carcinomas (BLC) but not in other types of 53 IDC (non-BLC) that were positive for ER, PgR and/or HER2. The percentage of CD109-positive tissues (60%) in BLC was similar to that of CK5/6 (63%) and higher than that of other myoepithelial markers including p63 (23%), calponin (33%) and vimentin (33%). Statistical analysis indicated that the CD109 positive group in BLC, but not the CK5/6-positive group in BLC, was associated with reduced fat invasion (P < 0.05). These findings indicate that CD109 is a useful diagnostic marker for BLC and that CD109 expression may affect biological properties of cancer cells. PMID- 18429828 TI - Galactosialidosis associated with IgA nephropathy: morphological study of renal biopsy. AB - Galactosialidosis is an autosomal recessive lysosomal disease associated with a deficiency of beta-galactosidase and neuraminidase. Described herein is the case of a young adult who had been diagnosed with galactosialidosis at 8 years of age. At the age of 30 years, proteinuria and hematuria appeared and the patient underwent a renal biopsy 1 year later. Light microscopy of the kidney sections indicated fine granular contents in the cytoplasm of glomerular endothelial and epithelial cells, arteriolar smooth muscles and proximal tubular epithelial cells on periodic acid silver-methenamin (PAM) stain. Electron microscopy of these cells indicated enlarged, smooth endoplasmic reticulum and lysosomes containing 150 nm-wide rods with a fine lattice structure at 66 A periodicity. Moreover, electron-dense deposits were located in the paramesangial area. Immunofluorescence staining indicated diffuse and global anti-human IgA and C3 positive staining as a mesangial pattern. Given these findings this patient was therefore diagnosed with both galactosialidosis and IgA nephropathy. This is the first report to describe light and electron microscopy observations of storage materials in the kidneys in young/adult galactosialidosis. PMID- 18429829 TI - Oncocytic renal cell carcinoma having papillotubular growth: rare morphological variant of papillary renal cell carcinoma. AB - Herein is described a unique renal cell tumor with previously unreported morphological and immunohistochemical features. The patient was a 78-year-old Japanese man. A huge left renal tumor was found on ultrasound during evaluation of left abdominal distention. The tumor was macroscopically characterized by a non-infiltrative border, pale yellow to grayish color, foci of hemorrhage and partial edematous change. Histologically the tumor had an extensive small tubular growth pattern often with papillary fronds mainly composed of oncocytic cells with deeply eosinophilic granular cytoplasm. Clear vacuolated cells were scattered among the oncocytic cells. The present case had an unusual immunohistochemical profile for all known types of renal cell tumors, but both the oncocytic cells and the clear vacuolated cells were strongly immunoreactive for alpha-methylacyl-coenzyme A racemase. It is concluded that the tumor may be a candidate for a rare variant of papillary renal cell carcinoma. Further cases having similar features are awaited for a definitive classification of this tumor as a previously undescribed tumor type. PMID- 18429830 TI - Sclerosing variant of epithelioid angiomyolipoma. AB - Presented herein are two unusual epithelioid angiomyolipomas (AML) displaying prominent stromal sclerosis. Both patients were middle-aged women without a clinical history of tuberous sclerosis. One patient (case 1) had a 2 cm lesion arising in the renal cortex, and another (case 2) had a pararenal retroperitoneal tumor measuring 13 cm. Both tumors were composed of sheets or nests of polygonal epithelioid or short spindle cells having uniform round to oval nuclei and eosinophilic cytoplasm with cords of hyalinized sclerotic stroma between them. The tumor in case 2 had small areas of mature-looking fat cells. Immunohistochemically, epithelioid tumor cells were diffusely positive for actins and desmin in both cases, and melanoma antigen recognized by T cells (MART)-1 was positive in patient 2. Scattered HMB-45-immunoreactive cells were identified in the sclerotic cords of both tumors, but epithelioid tumor cells were essentially negative for HMB-45. The characteristic clinicopathological and immunohistochemical features of the present cases are analogous to a subset of epithelioid AML or sclerosing perivascular epithelioid cell tumors previously reported. PMID- 18429831 TI - Serous borderline tumor of the paratestis. AB - Reported herein is a case of serous borderline tumor (SBT, ovarian epithelial type tumor) of the paratestis, involving the tunica vaginalis, in a 64-year-old man. The patient complained of right hydrocele; puncture cytology of the turbid fluid pointed to an adenocarcinoma. Right orchiectomy was performed and multiple micronodules were grossly observed in the paratestis. On microscopy small papillary epithelial lesions were found with psammoma bodies and intraglandular papillary lesions were irregularly recognized in the stroma of the paratestis, similar to SBT of the ovary. The tumor cells had often short microvilli. Mucin production was evident on PAS and colloid iron staining. Both papillary and glandular epithelial cells were positive on immunohistochemistry for Ber EP4/epithelial antigen, low-molecular-weight cytokeratin (CAM5.2), cytokeratin 7 and estrogen and progesterone hormone receptors, but negative for CEA, cytokeratin 20 and calretinin. The average proliferative index was approximately 10.5% as assessed on Ki-67 (MIB-1) staining. Ultrastructurally, the cells did not demonstrate any well-developed microvilli or secretory granules and immunohistochemical findings supported SBT of Mullerian type (ovarian epithelial type tumor), while excluding a papillary type of malignant mesothelioma. The lesion in the present case was concluded to be a testicular serous tumor of Mullerian type, similar to SBT of the ovary. PMID- 18429832 TI - Metastatic breast carcinoma identified in a uterine leiomyosarcoma. AB - A rare example is described of an advanced invasive ductal carcinoma of the breast with concomitant metastatic foci in a uterine leiomyosarcoma. Both tumors arose in a 48-year-old woman, without any evidence of distant metastases as assessed on imaging at the time of diagnosis. The double diagnosis in this particular case was established on a combination of microscopic appearance of the uterine tumors in the hysterectomy specimen, immunophenotype assessment, and clinical information on the breast carcinoma. But the association between the two malignancies can be challenging and potentially difficult to prove. PMID- 18429833 TI - Aggressive invasive micropapillary salivary duct carcinoma of the parotid gland. AB - The presence of invasive micropapillary component has been reported to be associated with salivary duct carcinoma and poor outcomes. Herein is described a rare case of invasive micropapillary salivary duct carcinoma of the parotid gland in a 60-year-old man. The micropapillary component was approximately 70% of the area of the tumor. Squamous differentiation was focally seen adjacent to the micropapillary component. On immunohistochemistry the ordinary salivary duct carcinoma component was positive for gross cystic disease fluid protein-15 (GCDFP 15), androgen receptor (AR), and HER2/neu, whereas both micropapillary and squamous components were negative for GCDFP-15 and AR. Immunohistochemical staining for D2-40 highlighted the lymph vessel invasion of tumor cells. This patient developed metastases in the lymph nodes of the neck, and also in the liver, lung, and brain. The lymph nodes and liver metastases had both ordinary salivary duct carcinoma and micropapillary components. The patient died of tumor 11 months after the initial surgical operation. The results support that the presence of micropapillary component is associated with more aggressive behavior of salivary duct carcinoma. It is also important for pathologists to recognize that GCDFP-15 and AR expression can be reduced in micropapillary carcinoma in the differential diagnosis of metastatic tumor. PMID- 18429834 TI - Looking forward to the next 20 years of JCAPN. PMID- 18429835 TI - Pediatric bipolar disorder: evidence-based psychopharmacological treatments. PMID- 18429836 TI - A model for caregiving of adopted children after institutionalization. PMID- 18429837 TI - Children in foster care: a vulnerable population at risk. AB - TOPIC: Nationally, 542,000 children are in foster care. Many of these children have prior histories of maltreatment such as abuse and neglect, with neglect being the most common form of maltreatment and the reason for many children requiring foster care services. Painful experiences associated with maltreatment and the trauma of being removed from one's parents (foster care) may affect the developmental and mental health of children. PURPOSE: This paper synthesizes the experiences associated with foster care and reveals foster care outcomes obtained through a literature search of published research. Specifically, the notions of oppression and domination defined by Young (1990) experienced by children in foster are explored. SOURCES: Review of the literature and clinical practice. CONCLUSIONS: Most children in foster care, if not all, experience feelings of confusion, fear, apprehension of the unknown, loss, sadness, anxiety, and stress. Such feelings and experiences must be addressed and treated early to prevent or decrease poor developmental and mental health outcomes that ultimately affect a child's educational experience and the quality of adulthood. Systemic orientation for all children entering foster care is proposed as a preventative intervention that addresses associated experiences of children in foster care. PMID- 18429838 TI - Exploring parent participation in a parent training program for children's aggression: understanding and illuminating mechanisms of change. AB - PROBLEM: Controlled trials have demonstrated that parents of children experiencing high levels of aggression benefit greatly from parent training programs. Several programs have shown a decrease in parental stress, an increase in parental confidence, and higher levels of prosocial behavior in children as shown by outcomes based on quantitative measures. However, less attention has been paid to the views and experiences of parents themselves about the impact of such programs on themselves, their children, and their parent-child relationships. METHODS: The purpose of this qualitative study was to elicit and explore parents' perceptions of the effectiveness of the Incredible Years Parent Training Program. Following their participation in the Incredible Years Program, 37 parents completed a semistructured interview and completed demographic questionnaires. Data were analyzed employing a content analysis of the transcripts and descriptive statistics of the demographic data. FINDINGS: Parents strongly valued the support offered within the group therapy process, reporting a decrease in their stress levels, an increase in their confidence, as well as observing positive changes in their children and in the parent-child relationship. CONCLUSIONS: The findings of this research illuminate possible underlying mechanisms for these observed changes. Specifically, when parents feel accepted, supported, and not blamed by healthcare professionals, they seem to be able to engage in self-reflection specifically related to their parenting styles. In turn, their ability to reflect in the group and make sense of their own thoughts, feelings, and behaviors seems to have a positive influence on the process of change in themselves, their children, and in their relationships with their children and other family members. PMID- 18429839 TI - What can we learn? Adult outcomes in children of seriously mentally ill mothers. AB - PROBLEM: Information is lacking about the experiences, needs of, and interventions for children of seriously mentally ill mothers. METHODS: Quantitative and qualitative methods of inquiry were used to retrospectively explore the characteristics and needs of adult children of seriously mentally ill mothers. The sample (N = 40) was recruited by referral and media advertisements. Childhood variables related to attachment, family environment, and parenting were compared to adult well-being outcomes of depression, quality of life, sense of coherence, and self-esteem. Participants also responded to the question "What other question should have been included in this study about your experience as the child of a seriously mentally ill mother?" and, additionally, spontaneously added their own clarifications of their answers to the survey questions. FINDINGS: It was apparent that the childhoods of participants were disruptive and often painful. Over half of the sample reported having their own diagnosis of depression in adulthood. Despite these factors, most members of the study sample were functioning well in adulthood, most often as a result of their own initiative. CONCLUSIONS: A high rate of depression in adulthood and participants' own descriptions of their painful memories and experiences of childhood identifies that more can and should be done to assist children of mentally ill mothers to cope with their environments. Interventions at various times in childhood are described. PMID- 18429841 TI - Role of nursing in juvenile justice. PMID- 18429840 TI - Korean American adolescent depression and parenting. AB - PROBLEM: Korean American adolescents tend to experience more mental health problems than adolescents in other ethnic groups. METHODS: The goal of this study was to examine the association between Korean American parent-adolescent relationships and adolescents' depressive symptoms in 56 families. FINDINGS: Thirty-nine percent of adolescents reported elevated depressive symptoms. Adolescents' perceived low maternal warmth and higher intergenerational acculturation conflicts with fathers were significant predictors for adolescent depressive symptoms. CONCLUSIONS: The findings can be used to develop a family intervention program, the aim of which would be to decrease adolescent depressive symptoms by promoting parental warmth and decreasing parent-adolescent acculturation conflicts. PMID- 18429842 TI - Kinship care and the situation for grandparents. PMID- 18429843 TI - A call for advocacy and action: national forum for mental health promotion and mental illness prevention. PMID- 18429844 TI - Making monsters: heterosexuality, crime and race in recent Western media coverage of HIV. AB - In the early HIV epidemic, Western media coverage encouraged the idea that infection was linked to 'other' identities located outside the 'mainstream'; outside 'proper' heterosexuality. Today, however, HIV has become repositioned as a global heterosexual epidemic. Analyses show that since the 1990s Western media have shifted away from blame and hysteria to an increasingly routinised reporting of HIV as a health story and social justice issue. But recent years have seen the emergence of a new media story in many Western countries; the criminal prosecution for HIV-related offences, and with it a reframing of old discourses of 'innocence' and 'guilt', but now with heterosexuals in focus. We examine this story in recent domestic media coverage in Australia, a country where heterosexual HIV transmission is rare by global comparison. Echoing similar stories in other Western media, in Australian coverage the idea of criminal intent converges with the symbolic weight of black sexuality and African origins to produce a 'monstrous' masculinity, which at the local level taps into contemporary racial tensions and, in so doing, conjures an imagined Anglo heterosexuality at once vulnerable to and safe from HIV in a globalised epidemic and world. PMID- 18429852 TI - A molecular genetic examination of the mating system of pumpkinseed sunfish reveals high pay-offs for specialized sneakers. AB - Intrasexual variation in reproductive behaviour and morphology are common in nature. Often, such variation appears to result from conditional strategies in which some individuals (e.g. younger males or those in poor condition) adopt a low pay-off phenotype as a 'best of a bad job'. Alternatively, reproductive polymorphisms can be maintained by balancing selection, with male phenotypes having equal fitnesses at equilibrium, but examples from nature are rare. Many species of sunfish (genus Lepomis) are thought to have alternative male reproductive behaviours, but most empirical work has focused on the bluegill sunfish and the mating systems of other sunfish remain poorly understood. We studied a population of pumpkinseed sunfish (Lepomis gibbosus) in upstate New York. Field observations confirm the existence of two male reproductive strategies: 'parentals' were relatively old and large males that maintained nests, and 'sneakers' were relatively young and small males that fertilize eggs by darting into nests of parentals during spawning. The sneaker and parental male strategies appear to be distinct life-history trajectories. Sneaker males represented 39% of the males observed spawning, and sneakers intruded on 43% of all mating attempts. Microsatellite analyses revealed that sneaker males fertilized an average of 15% of the eggs within a nest. This level of paternity by sneaker males appears to be higher than seen in most other fishes, and preliminary analyses suggest that the two male reproductive strategies are maintained as a balanced polymorphism. PMID- 18429853 TI - Metal exchange in metallothioneins: a novel structurally significant Cd(5) species in the alpha domain of human metallothionein 1a. AB - Metallothioneins (MTs) are cysteine-rich, metal-binding proteins known to provide protection against cadmium toxicity in mammals. Metal exchange of Zn(2+) ions for Cd(2+) ions in metallothioneins is a critical process for which no mechanistic or structural information is currently available. The recombinant human alpha domain of metallothionein isoform 1a, which encompasses the metal-binding cysteines between Cys33 and Cys60 of the alpha domain of native human metallothionein 1a, was studied. Characteristically this fragment coordinates four Cd(2+) ions to the 11 cysteinyl sulfurs, and is shown to bind an additional Cd(2+) ion to form a novel Cd(5)alpha-MT species. This species is proposed here to represent an intermediate in the metal-exchange mechanism. The ESI mass spectrum shows the appearance of charge state peaks corresponding to a Cd(5)alpha species following addition of 5.0 molar equivalents of Cd(2+) to a solution of Cd(4)alpha-MT. Significantly, the structurally sensitive CD spectrum shows a sharp monophasic peak at 254 nm for the Cd(5)alpha species in contrast to the derivative-shaped spectrum of the Cd(4)alpha-MT species, with peak maxima at 260 nm (+) and 240 nm (-), indicating Cd-induced disruption of the exciton coupling between the original four Cd(2+) ions in the Cd(4)alpha species. The (113)Cd chemical shift of the fifth Cd(2+) is significantly shielded (approximately 400 p.p.m.) when compared with the data for the Cd(2+) ions in Cd(4)alpha-MT by both direct and indirect (113)Cd NMR spectroscopy. Three of the four original NMR peaks move significantly upon binding the fifth cadmium. Evidence from indirect (1)H-(113)Cd HSQC NMR spectra suggests that the coordination environment of the additional Cd(2+) is not tetrahedral to four thiolates, as is the case with the four Cd(2+) ions in the Cd(4)alpha-MT, but has two thiolate ligands as part of its ligand environment, with additional coordination to either water or anions in solution. PMID- 18429854 TI - The association of psycho-social factors and survival in head and neck cancer. AB - OBJECTIVE: Update a previous review examining associations between psycho-social factors and survival in head and neck cancer patients. DATA SOURCES: Searched Cochrane, Psych info and Embase for the period from 1 January 1995 to 1 June 2007, as well as personal and article reference lists and article archives. STUDY SELECTION: Identified articles assessed by consensus for eligibility using following criteria: survival as outcome measure; psycho-social factors as prognostic indicators; results specifically for head and neck cancer patients, not including oesophageal or thyroid cancer. Seven of 64 articles fulfilled criteria. DATA EXTRACTION: Data abstracted independently by two reviewers using pre-determined proformas. Quality also rated using Scottish Intercollegiate Guidelines Network 50 tool. DATA SYNTHESIS: At baseline, expression of intense psycho-social complaints, higher self-perceived physical ability and self reported high physical functioning were significantly associated with increased survival. Uncertainty about the diagnosis and treatment was found to be a negative prognostic indicator, as was being single, poor cognitive function, baseline fatigue and alcoholism. Overall quality of life and head and neck pain 12 months after date of diagnosis were found to be significantly associated with survival in one study. However, overall quality of life and depression at the time of diagnosis were not. CONCLUSIONS: There appears to be some association between selected psycho-social factors and long-term survival from head and neck cancer. However this relationship is currently neither strong nor proven, requiring examination by multi centred trials with standardisation of research definitions and methodologies, and examination of post-treatment psycho-social factors. PMID- 18429855 TI - The evidence for different management strategies for laryngeal cancer: summaries of systematic reviews commissioned for an educational meeting. PMID- 18429856 TI - Chronic cough. PMID- 18429857 TI - A randomised controlled trial comparing fresh, dried, and dried-then-rehydrated temporalis fascia in myringoplasty. AB - OBJECTIVES: To determine whether the way in which temporalis fascia is treated during myringoplasty affects the rate of successful perforation closure. DESIGN: A randomised controlled trial. SETTING: A tertiary ENT Department within the Tygerberg Academic Hospital. PARTICIPANTS: One hundred and fifty patients with perforations between 30% and 70% of the surface area of the eardrum undergoing elective myringoplasty were randomised into three groups by the way in which the temporalis fascia used was treated prior to insertion: fresh fascia; dried fascia; and fascia which was dried and then rehydrated. MAIN OUTCOME MEASURES: The outcome measure was intactness of the tympanic membrane 6 weeks after surgery. Preoperative and follow-up audiometry was obtained, but was not evaluated as an outcome measure. RESULTS: Successful closure of the perforation was achieved in 89% in the dried fascia group (42 of 47 patients), 84% in the fresh fascia group (37 of 44 patients), and 85% in the dried and rehydrated group (39 of 46 patients). The study showed no statistically significant difference between the three groups (P = 0.728). The study would have to had more than 800 patients in each group to have the power to show a 5% difference in take rates. CONCLUSIONS: While the dessication of temporalis fascia prior to use in myringoplasy results in degeneration of cellular and stromal elements histologically, this does not affect the successful closure of perforations. Clinicians should feel free to prepare the fascia in whichever way they find easiest to work with. PMID- 18429858 TI - Identification of predictors and development of a screening protocol for cerebello-pontine lesions in patients presenting with audio-vestibular dysfunction. AB - OBJECTIVES: Magnetic resonance imaging (MRI) scanning in the investigation of cerebellopontine angle lesions represents a finite resource, the use of which needs to be carefully rationalised. Our aim was to identify predictive factors that can distinguish between patients with and without cerebellopontine angle lesions, and develop a screening protocol which could be useful in the clinical setting as an aid to clinical judgment. DESIGN: Case-control study. SETTING: Secondary care. PARTICIPANTS: Audio-vestibular features were collated on 136 patients (M : F 1.39 : 1) and 288 controls (M : F 1 : 1.1). INTERVENTION: Diagnostic by analysis of symptoms and audiometric data using logistic regression, receiver-operator characteristic curves and backwards elimination. MAIN OUTCOME MEASURES: Development of a predictive algorithm comprising those factors which are most strongly predictive of the presence of a cerebellopontine angle lesion. RESULTS: Positive predictors of a cerebellopontine angle lesion include the interaural threshold difference at 1 (P = 0.044) and 4 kHz (P = 0.034). The threshold in the better hearing ear at 0.25 kHz exerts a negative predictive (i.e. protective) effect (P = 0.005). The presence of tinnitus does not appear to influence the outcome on logistic regression. Although vertigo does exert an influence on the overall model, its impact is highly equivocal. CONCLUSIONS: We have identified audiometric factors which exert a positive and negative predictive influence on the presence of a cerebellopontine angle lesion, and audiovestibular symptoms which appear to exert little effect on the model. Our predictive equation represents a user-friendly standardised method of risk stratification of patients within a general otolaryngology clinic. PMID- 18429859 TI - Infant hearing screening: effects of timeline. AB - OBJECTIVES: Universal infant hearing screening using otoacoustic emission and auditory brain-stem response audiometry is widely administered to attain the goals of early identification of, and intervention for hearing impairment. Concerns regarding screening specificity have, however, been raised. False positives may result from vernix occlusion in the ear canal or transient middle ear effusion, and can result in substantial costs to health care systems. The current study investigates the effects of age and time interval between tests on hearing assessment results. SETTING & PARTICIPANTS: Three hundred and seventeen positive screens from a 2-stage distortion product otoacoustic emission (DPOAE) screening programme in Hong Kong, who subsequently received diagnostic auditory brainstem response (ABR) assessment and monitoring, were investigated. MAIN OUTCOME MEASURES: Differences in diagnostic ABR results were compared among infants of different ages at tests, and with different time lapses after DPOAE screening. The proportion of those having persistent hearing impairment, conductive loss and impairment of moderate degree or above, were also compared. RESULTS: A significantly higher rate of normal ABR thresholds (60%versus 24%) was noted in infants assessed after age 50 days, and in infants diagnostically assessed with a time lapse of over 20 days post-DPOAE screening (65%versus 42%). CONCLUSIONS: Delaying diagnostic ABR assessment may reveal a higher percentage of normal thresholds, and hence probably higher specificity. Time delay may allow for spontaneous resolution of transient outer and middle ear conditions. However, the goals of early identification and intervention, as well as possible parental anxiety with delayed assessment, should also be considered when reviewing infant hearing screening schedules. PMID- 18429863 TI - Ionomer cement for reconstruction of the long process of the incus: the Pecs experience. AB - Experience gained with the repair of the defects of the long process of the incus with ionomer cement in 35 patients is reported. The integrity of the reconstructed tympanic membrane and the four-frequency average of the air-bone gaps of the postoperative audiograms (best, most recent and at around 1 year) were evaluated. This 'physiological' reconstruction of the ossicular chain fulfilled our expectations in 40% of the patients (air-bone gap N670L671 mutations when compared with mock-transfected cells. Studies with fluorogenic substrates indicated that ABCA7 had no impact on alpha-, beta-, or gamma-secretase activities. Live cell imaging of Chinese hamster ovary cells expressing APP-GFP indicated an apparent retention of APP in a perinuclear location in ABCA7 co-transfected cells. These studies indicate that ABCA7 has the capacity to stimulate cellular cholesterol efflux to apoE discs and regulate APP processing resulting in an inhibition of Abeta production. PMID- 18429933 TI - VEGF concentration from plasma-activated platelets rich correlates with microvascular density and grading in canine mast cell tumour spontaneous model. AB - Canine cutaneous mast cell tumour (CMCT) is a common cutaneous tumour in dog, with a higher incidence than in human. CMCT is classified in three subgroups, well and intermediately differentiated (G1 and G2), corresponding to a benign disease, and poorly differentiated (G3), corresponding to a malignant disease, which metastasize to lymph nodes, liver, spleen and bone marrow. In this study, we have evaluated serum (S), platelet-poor plasma (P-PP), plasma-activated platelet rich (P-APR) and cytosol vascular endothelial growth factor (VEGF) concentrations, microvascular density (MVD) and mast cell density (MCD) in a series of 86 CMCTs and we have correlated these parameters with each other, by means of ELISA detection of VEGF and immunohistochemistry. Results show that VEGF level from cytosol P-APR and MVD were significantly higher in G3 CMCTs as compared to G1 or G2 subgroups. Moreover, a significantly strong correlation among VEGF levels from P-PAR and cytosol, MVD and MCD was found in G3 subgroup. Because VEGF levels from P-APR well correlated with MVD and malignancy grade in CMCT, we suggest that VEGF might be secreted from MCs and it may be a suitable surrogate inter-species angiogenetic markers of tumour progression in CMCT. Finally, CMCT seems to be a useful model to study the role of MCs in tumour angiogenesis and inhibition of MCs degranulation or activation might be a new anti-angiogenic strategy worthy to further investigations. PMID- 18429934 TI - Angiogenic transforming capacity of IgG purified from plasma of type 1 diabetic patients. AB - We previously demonstrated that plasma of type 1 diabetic patients contains antibodies complexed irreversibly with Grp94 that also display proteolytic activity. In this work, we wanted to test whether antibodies obtained from diabetic plasma may convey an inflammatory risk on vascular cells. To this aim, IgG were purified on the Protein-G column from individual plasma of eight type 1 diabetic patients, and then tested on HUVECs to measure effects on cell growth and morphologic changes at different incubation times. The purified fractions of IgG contained a significant amount of Fab/(Fab)(2), both free and in big aggregates, and anti-Grp94 antibodies, mostly irreversibly linked with, but also free of Grp94. The purified fractions of both Fab/(Fab)(2) and whole IgG stimulated the proliferation and sustained the angiogenic differentiation of human umbilical vein endothelial cells (HUVECs) at sub-nanomolar concentrations. IgG from normal plasma neither stimulated the cell growth nor induced any differentiation of HUVECs. The maximum cell growth stimulation occurred at 6-9 hrs and associated with the strong activation of the ERK1/2 pathway, whereas angiogenic transformation was completed later when the ERK1/2 activation was silenced and cell growth stimulation significantly reduced. Neither proteolytic activity of MMP-9 nor VEGF were apparently involved in mediating the angiogenic differentiation of HUVECs that mostly correlated with an increased expression of HSP70 closely coupled with cell membrane-bound inactive species of MMP-9. Results indicate that effects displayed on HUVECs by antibodies purified from diabetic plasma are likely sustained by immune complexes with Grp94 that may thus predict an increased risk of angiogenic transformation in vivo. PMID- 18429935 TI - Adelmidrol, a palmitoylethanolamide analogue, reduces chronic inflammation in a carrageenin-granuloma model in rats. AB - Palmitoylethanolamide (PEA) and some of its analogues have shown great efficacy in the treatment of pain and inflammation. Adelmidrol - the International Nonproprietary Name (INN) of the di-amide derivative of azelaic acid - is one of these analogues. The anti-inflammatory and analgesic effects of PEA and adelmidrol are hypothesized to be mediated, at least in part, by mast cell down modulation. Mast cell mediators released at early stage of the inflammatory process drive the inflammatory reaction to chronicity as it happens in X carrageenin-induced granulomatous tissue formation. In the present study, the choice of testing adelmidrol depends upon the physicochemical properties of the compound, i.e. the amphipatic feature, that make it more easily soluble than PEA. In this study, we investigated the effect of adelmidrol on granuloma formation induced by lambda-carrageenin-soaked sponge implant in rats. Our results show that the local administration of the compound under study significantly decreases weight and neo-angiogenesis in granulomatous tissue. The anti-inflammatory effect was due to the modulation of mast cells degranulation, as shown by histological analysis and by the inhibition of the release of several pro-inflammatory and pro angiogenic enzymes (e.g. iNOS, chymase and metalloproteinase MMP-9), and mediators (e.g. nitric oxide and TNF-alpha). The results indicate that adelmidrol, given locally, may represent a potential therapeutic tool in controlling chronic inflammation. PMID- 18429937 TI - Deletion of the chloroplast-localized AtTerC gene product in Arabidopsis thaliana leads to loss of the thylakoid membrane and to seedling lethality. AB - Early seedling development in plants depends on the biogenesis of chloroplasts from proplastids, accompanied by the formation of thylakoid membranes. An Arabidopsis thaliana gene, AtTerC, whose gene product shares sequence similarity with bacterial tellurite resistance C (TerC), is shown to be involved in a critical step required for the normal organization of prothylakoids and transition into mature thylakoid stacks. The AtTerC gene encodes an integral membrane protein, which contains eight putative transmembrane helices, localized in the thylakoid of the chloroplast, as shown by localization of an AtTerC-GFP fusion product in protoplasts and by immunoblot analysis of subfractions of chloroplasts. T-DNA insertional mutation of AtTerC resulted in a pigment deficient and seedling-lethal phenotype under normal light conditions. Transmission electron microscopic analysis revealed that mutant etioplasts had normal prolamellar bodies (PLBs), although the prothylakoids had ring-like shapes surrounding the PLBs. In addition, the ultrastructures of mutant chloroplasts lacked thylakoids, did not have grana stacks, and showed numerous globular structures of varying sizes. Also, the accumulation of thylakoid membrane proteins was severely defective in this mutant. These results suggest that the AtTerC protein plays a crucial role in prothylakoid membrane biogenesis and thylakoid formation in early chloroplast development. PMID- 18429938 TI - Inter-dependence of dimerization and organelle binding in myosin XI. AB - Cytoplasmic streaming is a ubiquitous process in plant cells that is thought to be driven by the active movement of myosin XI motor proteins along actin filaments. These myosin motors bind to organelles through their C-terminal globular tail domain, although recent studies have also suggested a role for the central coiled-coil region during organelle binding. Here we have investigated the relationship between these two protein domains of MYA1, an Arabidopsis myosin XI, in a series of in vivo experiments demonstrating that dimerization of the coiled-coil region stabilizes organelle binding of the globular tail. Surprisingly, yeast two-hybrid assays, bimolecular fluorescence complementation, Forster resonance energy transfer and in vitro pull-down experiments all demonstrated that dimerization of the 174-residue MYA1 coiled coils by themselves was unstable. Furthermore, only the first of the two major coiled-coil segments in MYA1 contributed significantly to dimer formation. Interestingly, dimerization of myosin tail constructs that included the organelle-binding globular tail was stable, although the globular tails by themselves did not interact. This suggests an inter-dependent relationship between dimerization and organelle binding in myosin XI, whereby each process synergistically stimulates the other. PMID- 18429936 TI - Roles of interstitial cells of Cajal in regulating gastrointestinal motility: in vitro versus in vivo studies. AB - The aim of this article is to provide a better understanding of the roles of interstitial cells of Cajal (ICC) in regulating gastrointestinal motility by reviewing in vitro and in vivo physiological motility studies. Based on the in vitro studies, ICC are proposed to have the following functions: to generate slow waves, to mediate neurotransmission between the enteric nerves and the gastrointestinal muscles and to act as mechanoreceptors. However, there is limited evidence available for these hypotheses from the in vivo motility studies. In this review, we first introduce the major subtypes of ICC and their established functions. Three Kit mutant mouse and rodent models are presented and the loss of ICC subtypes in these mutants is reviewed. The physiological motility findings from various in vitro and in vivo experiments are discussed to give a critical review on the roles of ICC in generating slow waves, regulating gastrointestinal motility, mediating neural transmission and serving as mechanoreceptors. It is concluded that the role of ICC as pacemakers may be well established, but other cells may also be involved in the generation of slow waves; the theory that ICC are mediators of neurotransmission is challenged by the majority of the in vivo motility studies; the hypothesis that ICC are mechanoreceptors has not found supportive evidence from the in vivo studies yet. More studies are needed to explain discrepancies in motility findings between the in vitro and in vivo experiments. PMID- 18429939 TI - The eer5 mutation, which affects a novel proteasome-related subunit, indicates a prominent role for the COP9 signalosome in resetting the ethylene-signaling pathway in Arabidopsis. AB - An Arabidopsis mutant, eer5-1, which has an enhanced ethylene response in etiolated seedlings, including hypersensitivity and extreme exaggeration of response to ethylene, was isolated and characterized. As with other identified eer mutants, the enhanced response phenotype of eer5-1 was correlated with failure to induce appropriately a subset of ethylene-regulated genes, suggesting that proper ethylene-responsive gene expression is necessary for resetting the ethylene response pathway. eer5-1 represents a mutation that causes an amino acid substitution in a previously uncharacterized gene, which encodes a protein with a PAM [proteasome COP9 initiation factor (PCI/PINT)-associated module] domain similar to those found in components of the COP9 signalosome (CSN). Genetic analysis shows that manifestation of the eer5 mutant phenotype is solely dependent on ethylene signaling, as the ein2-5 eer5-1 double mutant was indistinguishable from ein2-5 in the presence of saturating ethylene concentrations. In contrast, the ein3-1 eer5-1 double mutant displayed characteristics of an enhanced ethylene response, and this suggests that EER5 regulates ethylene signaling independently of EIN3. Analysis of the EER5 protein indicates that it interacts with the C-terminus of EIN2 and with the CSN, suggesting that EER5 serves as a bridge between EIN2 and the modification or degradation of target proteins, including a proposed group of transcriptional repressors, as part of a resetting mechanism during or following ethylene signaling. PMID- 18429940 TI - Enzymatic characterization of peroxisomal and cytosolic betaine aldehyde dehydrogenases in barley. AB - Betaine aldehyde dehydrogenase (BADH; EC 1.2.1.8) is an important enzyme that catalyzes the last step in the synthesis of glycine betaine, a compatible solute accumulated by many plants under various abiotic stresses. In barley (Hordeum vulgare L.), we reported previously the existence of two BADH genes (BBD1 and BBD2) and their corresponding proteins, peroxisomal BADH (BBD1) and cytosolic BADH (BBD2). To investigate their enzymatic properties, we expressed them in Escherichia coli and purified both proteins. Enzymatic analysis indicated that the affinity of BBD2 for betaine aldehyde was reasonable as other plant BADHs, but BBD1 showed extremely low affinity for betaine aldehyde with apparent K(m) of 18.9 microM and 19.9 mM, respectively. In addition, V(max)/K(m) with betaine aldehyde of BBD2 was about 2000-fold higher than that of BBD1, suggesting that BBD2 plays a main role in glycine betaine synthesis in barley plants. However, BBD1 catalyzed the oxidation of omega-aminoaldehydes such as 4-aminobutyraldehyde and 3-aminopropionaldehyde as efficiently as BBD2. We also found that both BBDs oxidized 4-N-trimethylaminobutyraldehyde and 3-N-trimethylaminopropionaldehyde. PMID- 18429941 TI - Gravisensitivity and automorphogenesis of lentil seedling roots grown on board the International Space Station. AB - The GRAVI-1 experiment was brought on board the International Space Station by Discovery (December 2006) and carried out in January 2007 in the European Modular Cultivation System facility. For the first run of this experiment, lentil seedlings were hydrated and grown in microgravity for 15 h and then subjected for 13 h 40 min to centrifugal accelerations ranging from 0.29 x 10(-2) g to 0.99 x 10(-2) g. During the second run, seedlings were grown either for 30 h 30 min in microgravity (this sample was the control) or for 21 h 30 min and then subjected to centrifugal accelerations ranging from 1.2 x 10(-2) g to 2.0 x 10(-2) g for 9 h. In both cases, root orientation and root curvature were followed by time-lapse photography. Still images were downlinked in near real time to ground Norwegian User Support and Operations Center during the experiment. The position of the root tip and the root curvature were analyzed as a function of time. It has been shown that in microgravity, the embryonic root curved strongly away from the cotyledons (automorphogenesis) and then straightened out slowly from 17 to 30 h following hydration (autotropism). Because of the autotropic straightening of roots in microgravity, their tip was oriented at an angle close to the optimal angle of curvature (120 degrees -135 degrees ) for a period of 2 h during centrifugation. Moreover, it has been demonstrated that lentil roots grown in microgravity before stimulation were more sensitive than roots grown in 1 g. In these conditions, the threshold acceleration perceived by these organs was found to be between 0 and 2.0 x 10(-3) g and estimated punctually at 1.4 x 10(-5) g by using the hyperbolic model for fitting the experimental data and by assuming that autotropism had no or little impact on the gravitropic response. Gravisensing by statoliths should be possible at such a low level of acceleration because the actomyosin system could provide the necessary work to overcome the activation energy for gravisensing. PMID- 18429942 TI - Redox signal integration: from stimulus to networks and genes. AB - Recent research has established redox-dependent thiol modification of proteins as a major regulatory layer superimposed on most cell functional categories in plants. Modern proteomics and forward as well as reverse genetics approaches have enabled the identification of a high number of novel targets of redox regulation. Redox-controlled processes range from metabolism to transport, transcription and translation. Gene activity regulation by transcription factors such as TGA, Athb 9 and RAP2 directly or indirectly is controlled by the redox state. Knowledge on putative redox sensors such as the peroxiredoxins, on redox transmitters including thioredoxins and glutaredoxins and biochemical mechanisms of their linkage to the metabolic redox environment has emerged as the framework of a functional redox regulatory network. Its basic principle is similar in eukaryotic cells and particularly complex in the photosynthesizing chloroplast. Methods and knowledge are now at hand to develop a quantitative understanding of redox signalling and the redox regulatory network in the eukaryotic cell. PMID- 18429943 TI - Circulating endothelial cells in health and disease: how do we best quantify them? PMID- 18429944 TI - Prevention of venous thromboembolism in patients with immobilization of the lower extremities: a meta-analysis of randomized controlled trials. AB - BACKGROUND: It is controversial whether the prevention of venous thromboembolism (VTE) in patients with lower-leg immobilization is necessary. OBJECTIVES: To assess the benefits and complications of pharmacological thromboprophylaxis, we performed a meta-analysis of all available randomized controlled trials on this subject. METHODS: We searched the MEDLINE and EMBASE electronic databases. We included English- and non-English language studies, we hand-searched journals, and we contacted manufacturers. We analyzed data from six randomized trials that investigated pharmacological prophylaxis for the prevention of VTE in 1456 patients with lower-leg immobilization in plaster cast or orthosis. Two reviewers independently assessed the trials for inclusion, extracted data, and assessed trial quality. Differences were resolved by consensus or arbitrage. RESULTS: The pooled estimate from all trials revealed a highly significant and clinically relevant reduction in asymptomatic events with low-molecular-weight heparin (LMWH) prophylaxis, compared to placebo or untreated control [risk ratio (RR) 0.58; 95% confidence interval (CI) 0.39-0.86; P = 0.006). The mean rate of VTE decreased from 17.1% to 9.6% with the use of LMWH. Subgroup analysis of methodologically superior trials (RR 0.68; CI 0.50-0.92; P = 0.01), proximal deep vein thrombosis (RR 0.28; CI 0.11-0.72; P = 0.008), tendon ruptures (RR 0.60; CI 0.38-0.97; P = 0.04), and fractures (RR 0.62; CI 0.45-0.86; P = 0.004) confirmed the robustness of the overall result. Frequency of bleeding did not differ between LMWH prophylaxis and control groups (RR 1.22; CI 0.61-2.46; P = 0.57). CONCLUSIONS: Our findings indicate that thromboprophylaxis with LMWH for immobilization of the lower extremities reduces the risk of VTE. This benefit is achieved with no excess bleeding. PMID- 18429947 TI - Endocrine and metabolic disorders in young adult survivors of childhood acute lymphoblastic leukaemia (ALL) or non-Hodgkin lymphoma (NHL). AB - BACKGROUND: Treatments of acute lymphoblastic leukaemia (ALL) and non-Hodgkin lymphoma (NHL), involving various combinations of chemotherapy (chemo), cranial irradiation (CI) and/or bone marrow transplantation after total body irradiation (BMT/TBI), are often successful but may have several long-term harmful effects. OBJECTIVE: To evaluate late endocrine and metabolic complications in adult survivors of childhood ALL and NHL, in relation with the different therapeutic schemes received. DESIGN: Endocrine and metabolic parameters were determined in 94 patients (48 men, mean age: 24 +/- 5 years) with a former childhood ALL (n = 78) or NHL (n = 16) and subgrouped according to their previous treatment: chemo only (group I; n = 44), chemo + CI (group II; n = 32) and chemo + BMT/TBI (group III; n = 18). RESULTS: Severe GH deficiency (peak < 3.0 ng/ml after glucagon) was observed in 22% and 50% of patients of groups II and III, respectively, while hypothyroidism was mainly observed in group III (56%). Moreover, 83% of men developed hypogonadism after BMT/TBI, compared to 17% and 8% in groups I and II, respectively (P < 0.05), and all grafted women had ovarian failure, in contrast with other female patients in whom menarche had occurred spontaneously. Patients with BMT/TBI had also an adverse metabolic profile, with insulin resistance in 83% and dyslipidaemia in 61%. CONCLUSIONS: This study reveals a high prevalence of endocrine and metabolic disorders in young adult survivors of childhood ALL or NHL, this frequency mainly depending on the treatment received. Treatment with BMT/TBI is the most detrimental and many of these patients will develop GHD, hypothyroidism, hypogonadism, insulin resistance and dyslipidaemia. PMID- 18429948 TI - Effects of T4 replacement therapy on glucose metabolism in subjects with subclinical (SH) and overt hypothyroidism (OH). AB - OBJECTIVE: To evaluate beta-cell function and insulin sensitivity in subjects with overt (OH) and subclinical hypothyroidism (SH) before and after T4 replacement therapy. BACKGROUND: Disturbances in glucose metabolism have been observed in hypothyroid states. However, the clinical significance and potential reversibility of these alterations by T4 replacement therapy remain to be elucidated especially in patients with SH. DESIGN AND PATIENTS: Parameters of glucose metabolism have been investigated in subjects with OH (n = 12) and SH (n = 11). Insulin sensitivity has been assessed by the euglycaemic-hyperinsulinaemic clamp technique and beta-cell function by mathematical modelling of data derived from an oral glucose tolerance test. RESULTS: Fasting and dynamic glycaemia as assessed by the AUC(Glucose) remained unaltered following substitution therapy (P > 0.05). Insulin sensitivity significantly improved only in subjects with OH (P < 0.05). Fasting insulin and proinsulin concentrations increased proportionally in both groups (P < 0.05) with the proinsulin : insulin ratio remaining unchanged (P > 0.05). Total insulin secretion was higher in OH before initiation of therapy (P < 0.05). In both groups, dynamic parameters including total insulin secretion, hepatic insulin extraction and the adaptation index were significantly attenuated (P < 0.05) after restoration of thyroid function, whereas the disposition index and the basal insulin secretion rate remained unaltered (P > 0.05). CONCLUSION: In summary, SH and OH are characterized by attenuated basal plasma insulin levels and increased glucose-induced insulin secretion. T4 replacement therapy partially ameliorates the insulin secretion profile and reduces the demand on pancreatic beta-cells after glucose challenge to an extent that exceeds any effect attributable to the improvement in insulin sensitivity. PMID- 18429949 TI - Radioiodine therapy (RAI) for Graves' disease (GD) and the effect on ophthalmopathy: a systematic review. AB - BACKGROUND: An association between radioiodine therapy (RAI) for Graves' disease (GD) and the development or worsening of Graves' ophthalmopathy (GO) is widely quoted but there has been no systematic review of the evidence. AIMS: We undertook a systematic review of randomized controlled trials (RCTs) to assess whether RAI for GD is associated with increased risk of ophthalmopathy compared with antithyroid drugs (ATDs) or surgery. We also assessed the efficacy of glucocorticoid prophylaxis in the prevention of occurrence or progression of ophthalmopathy, when used with RAI. METHODS: We identified RCTs regardless of language or publication status by searching six databases and trial registries. Dual, blinded data abstraction and quality assessment were undertaken. Random effects meta-analyses were used to combine the study data. Ten RCTs involving 1136 patients permitted 13 comparisons. Two RCTs compared RAI with ATD. Two RCTs compared RAI with thyroidectomy. Four RCTs compared the use of adjunctive ATD with RAI vs. RAI. Five RCTs examined the use of glucocorticoid prophylaxis with RAI. RESULTS: RAI was associated with an increased risk of ophthalmopathy compared with ATD [relative risk (RR) 4.23; 95% confidence interval (CI): 2.04 8.77] but compared with thyroidectomy, there was no statistically significant increased risk (RR 1.59, 95% CI 0.89-2.81). The risk of severe GO was also increased with RAI compared with ATD (RR 4.35; 95% CI 1.28-14.73). Prednisolone prophylaxis for RAI was highly effective in preventing the progression of GO in patients with pre-existing GO (RR 0.03; 95% CI 0.00-0.24). The use of adjunctive ATD with RAI was not associated with any significant benefit on the course of GO. CONCLUSION: RAI for GD is associated with a small but definite increased risk of development or worsening of Graves' ophthalmopathy compared with ATDs. Steroid prophylaxis is beneficial for patients with pre-existing GO. PMID- 18429950 TI - Expression of mRNA for specific fibroblast growth factors associates with that of the myogenic markers MyoD and proliferating cell nuclear antigen in regenerating and overloaded rat plantaris muscle. AB - AIM: To examine the relations between specific fibroblast growth factors (FGFs) and satellite cell activation during muscle regeneration and hypertrophy in vivo, we measured mRNA expression of FGFs and myogenic markers in rat plantaris muscle after bupivacaine administration and synergist ablation. METHODS: mRNA levels for MyoD, myogenin, proliferating cell nuclear antigen (PCNA), p21, M-cadherin, Pax7, FGF-1, FGF-2, FGF-3, FGF-4, FGF-5, FGF-6, FGF-7, FGF-8 and hepatocyte growth factor (HGF) were measured continually for up to 72 h after bupivacaine administration and synergist ablation. FGF-5, FGF-7 and HGF proteins were immunostained at 72 h after bupivacaine administration. RESULTS: MyoD and PCNA mRNAs started increasing 24 h after bupivacaine administration. Myogenin, p21, M cadherin and Pax7 mRNAs started to increase after 48 and 72 h. After synergist ablation, MyoD, PCNA, M-cadherin and Pax7 mRNAs had increased at 24 and 48 h, and myogenin and p21 mRNAs at 12 and 24 h. FGF-1, FGF-7 and HGF mRNAs after the treatments started to increase at the same time as MyoD and PCNA mRNAs. FGF-5 was expressed at the same time as MyoD and PCNA mRNAs after bupivacaine administration but did not after the ablation. FGF-2, FGF-3, FGF-4, FGF-6 and FGF 8 mRNAs were not associated with the expression of the myogenic markers. FGF-7 and HGF proteins were expressed in immature muscle fibre nuclei and the extracellular matrix, but FGF-5 protein was preferentially expressed in extracellular matrix. CONCLUSION: These results indicate that FGF-1, FGF-7 and HGF are associated with specific myogenic marker expression during muscle regeneration and hypertrophy. PMID- 18429951 TI - Endosomal sorting complex required for transport proteins in cancer pathogenesis, vesicular transport, and non-endosomal functions. AB - Endosomal sorting complex required for transport (ESCRT) proteins form a multicomplex sorting machinery that controls multivesicular body (MVB) formation and the sorting of ubiquitinated membrane proteins to the endosomes. Being sorted to the MVB generally results in the lysosome-dependent degradation of cell surface receptors, and defects in this machinery induce dysregulated receptor traffic and turnover. Recent lessons from gene targeting and silencing methodologies have implicated the ESCRT in normal development, cell differentiation, and growth, as well as in the budding of certain enveloped viruses. Furthermore, it is becoming apparent that the dysregulation of ESCRT proteins is involved in the development of various human diseases, including many types of cancers and neurodegenerative disorders. Here, we summarize the roles of ESCRT proteins in MVB sorting processes and the regulation of tumor cells, and we discuss some of their other functions that are unrelated to vesicular transport. PMID- 18429953 TI - Age-related Epstein-Barr virus-associated B-cell lymphoproliferative disorders: special references to lymphomas surrounding this newly recognized clinicopathologic disease. AB - Epstein-Barr virus (EBV) is associated with some disease entities of malignant lymphomas, including Burkitt lymphoma, Hodgkin lymphoma, immunodeficiency associated lymphoproliferative disorders (LPD), and a part of diffuse large B cell lymphoma. We have recently identified a series of elderly patients with EBV associated (or EBV(+)) B-cell LPD (B-LPD) showing similarities in many respects to immunodeficiency-associated LPD, although no evidence of underlying immunodeficiency was found. Therefore, the nosological category of senile or age related EBV(+) B-LPD has been proposed for those patients. A larger series of patients with this disease revealed that the relative ratios of such EBV(+) B-LPD to all diffuse large B-cell lymphoma cases were higher with increasing with age, reaching a peak (20-30%) at > or =90 years of age, with a median of 71 years, providing additional evidence for our assertion that this disease may be related to immunological deterioration as a result of the aging process. This new disease entity is characterized pathologically by centroblasts, immunoblasts, and Hodgkin and Reed-Sternberg-like giant cells with a varying degree of reactive components, often posing therapeutic and diagnostic problems for hematologists and pathologists, respectively. The aim of the present review is to briefly summarize the overall clinicopathological profile of this newly recognized age-related (also called 'senile') EBV(+) B-LPD and EBV(+) Hodgkin lymphoma for a practical diagnostic approach. PMID- 18429952 TI - Human homolog of NOTUM, overexpressed in hepatocellular carcinoma, is regulated transcriptionally by beta-catenin/TCF. AB - The Drosophila Notum gene, which is regulated by the Wingless pathway, encodes a secreted hydrolase that modifies heparan sulfate proteoglycans. In comparative analysis of the gene expression profiles in primary human hepatocellular carcinomas (HCC) and normal organs, we observed that the human ortholog of Drosophila Notum was overexpressed markedly in a subset of HCC, but expressed rarely in adult normal tissues. Immunoblotting confirmed the overexpression of NOTUM protein in 12 of 40 primary HCC cases (30%). High levels of NOTUM protein were significantly associated with intracellular (nuclear or cytoplasmic) accumulation of beta-catenin protein: all 10 HCC with high intracellular beta catenin also had high NOTUM expression, whereas only 2 of 30 cases (6.7%) without intracellular beta-catenin had high NOTUM expression (P < 0.00001). NOTUM expression in HepG2 cells was downregulated significantly by induction of a dominant-negative mutant of TCF4, a beta-catenin partner. In vivo binding of the beta-catenin/TCF complex to the NOTUM promoter was demonstrated by chromatin immunoprecipitation in HepG2 and SW480 cells, where canonical Wnt signaling is activated constitutively. These findings provide evidence that NOTUM is a novel target of beta-catenin/TCF4 and is upregulated in Wnt/beta-catenin signaling activated HCC. PMID- 18429954 TI - Soy consumption reduces the risk of non-small-cell lung cancers with epidermal growth factor receptor mutations among Japanese. AB - Epidermal growth factor receptor (EGFR) mutations play substantial roles in genesis and proliferation of non-small-cell lung cancers (NSCLCs). We recently found that reproductive factors have a substantial impact on risk of development of NSCLCs featuring such EGFR mutations. Therefore, we explored the influence of dietary habits on NSCLC risk with reference to the EGFR mutational status. We conducted a case-control study using 353 patients with NSCLCs (122 EGFR mutated and 231 EGFR wild-type) and 1765 age-sex matched non-cancer control subjects. Dietary exposure was based on a semiquantitative food frequency questionnaire and impact of major food items, like meats, seafoods, vegetables and soybean products was assessed by multivariate logistic regression. Soybean products demonstrated a protective association with EGFR mutated, but not EGFR wild-type NSCLCs, with multivariate-adjusted odds ratios and 95% confidence intervals for the 2nd and 3rd tertile of soybean product consumption of 0.79 (0.50-1.27) and 0.56 (0.34 0.93) relative to those in the lowest tertile (trend P = 0.023). In conclusion, soy consumption may exert a protective association against the development of NSCLCs with EGFR mutations, providing possible insights into mechanisms of their genesis. PMID- 18429955 TI - Enhanced radiation response of a solid tumor with the artificial oxygen carrier 'albumin-heme'. AB - Tumor-cell hypoxia is one of the main factors inducing radioresistance. Enhanced tumor oxygenation has previously been achieved in an animal model using the synthetic heme-based oxygen carrier 'albumin-heme' (recombinant human serum albumin-Fe cyclohexanoil heme; rHSA-FeP). The present study was done to determine whether rHSA-FeP enhances the radiation response in an experimental tumor model. Male Donryu rats and LY80, a variant of the syngenic liver ascites tumor, were used. A total of 1 x 10(6) cells were injected into the subfascial tissue of the right thigh. The rats were divided randomly into five groups: sham (tumor implantation and sham operation); rHSA-FeP; irradiation; rHSA + irradiation; and rHSA-FeP + irradiation. Six days after, under general anesthesia, intra-arterial administration of 10 mL/kg of either 5% rHSA solution or oxygenated rHSA-FeP solution at 2.5 mL/min was done and a dose of 20 Gy was given. There were significant differences in tumor growth between the sham and irradiation groups, and between the sham and rHSA-FeP + irradiation groups. Tumor growth delay was observed and differences were significant between the sham and irradiation groups, and between the irradiation and rHSA-FeP + irradiation groups. In the present study, rHSA-FeP itself had a slight effect on tumor growth without irradiation. Enhancing the effect of rHSA-FeP on the radiation response is responsible in part for the oxygen-carrying property of rHSA-FeP. In conclusion, rHSA-FeP is a candidate radiation-enhancing drug. Arterial infusion of rHSA-FeP may serve as a local oxygenation method that enhances the radiation effect. PMID- 18429956 TI - Activity of a novel Aurora kinase inhibitor against the T315I mutant form of BCR ABL: in vitro and in vivo studies. AB - Despite promising results from clinical studies of ABL kinase inhibitors, a challenging problem that remains is the T315I mutation against which neither nilotinib nor dasatinib show significant activity. In the present study, we investigated the activity of a novel Aurora kinase inhibitor, VE-465, against leukemia cells expressing wild-type BCR-ABL or the T315I mutant form of BCR-ABL. We observed a dose-dependent reduction in the level of BCR-ABL autophosphorylation in VE-465-treated cells. Exposure to the combination of VE 465 and imatinib exerted an enhanced apoptotic effect in K562 cells. Combined treatment with VE-465 and imatinib caused more attenuation of the levels of phospho-AKT and c-Myc in K562 cells. Further, the isobologram indicated the synergistic effect of simultaneous exposure to VE-465 and imatinib in K562 cells. To assess the in vivo efficacy of VE-465, athymic nude mice were injected intravenously with BaF3 cells expressing wild-type BCR-ABL or the T315I mutant form. The vehicle-treated mice died of a condition resembling acute leukemia by 28 days; however, nearly all mice treated with VE-465 (75 mg/kg, twice daily; intraperitoneally for 14 days) survived for more than 56 days. Histopathological analysis of vehicle-treated mice revealed infiltration of the spleen. In contrast, histopathological analysis of organs from VE-465-treated mice demonstrated normal tissue architecture. Taken together, the present study shows that VE-465 exhibits a desirable therapeutic index that can reduce the in vivo growth of T315I mutant form and wild-type BCR-ABL-expressing cells in an efficacious manner. PMID- 18429957 TI - Resveratrol induces apoptosis in K562 (chronic myelogenous leukemia) cells by targeting a key survival protein, heat shock protein 70. AB - Chronic myelogenous leukemia (CML) is a myeloproliferative disease associated with a characteristic chromosomal translocation called the Philadelphia chromosome. This results in the expression of the Bcr-Abl fusion protein, a constitutively active protein tyrosine kinase. Although there are a few treatment options with Bcr-Abl kinase inhibitors, drug resistance is often encountered. One of the major obstacles in overcoming drug resistance in CML is the high endogenous levels of heat shock protein 70 (Hsp70). Resveratrol is a phytoalexin produced by several plants. We studied the chemotherapeutic effects and mode of action of resveratrol on K562 (CML) cells. Resveratrol induced apoptosis in K562 cells in a time-dependent manner. This was established by increased annexin V binding, corroborated with an enhanced caspase-3 activity and a rise in the sub G(0)/G(1) population. Resveratrol treatment also caused suppression of Hsp70 both in mRNA and protein levels. The downregulation of Hsp70 by resveratrol exposure was correlated with a diminished presence of heat shock factor 1 (HSF1) in the nucleus, and the downregulation of transcriptional activity of HSF1. High endogenous levels of Hsp70 have been found to be a deterrent for sensitivity to chemotherapy. We show here that resveratrol could considerably enhance the apoptosis induction in K562 cells by 17-allylamino-17-demethoxygeldanamycin, an anticancer agent that inhibits Hsp90 but augments Hsp70 levels. We conclude that resveratrol significantly downregulated Hsp70 levels through inhibition of HSF1 transcriptional activity and appreciably augmented the pro-apoptotic effects of 17-allylamino-17-demethoxygeldanamycin. PMID- 18429958 TI - Suicide gene/prodrug therapy using salmonella-mediated delivery of Escherichia coli purine nucleoside phosphorylase gene and 6-methoxypurine 2'-deoxyriboside in murine mammary carcinoma 4T1 model. AB - Attenuated salmonella have been reported to be capable of both selectively growing in tumors and expressing exogenous genes for tumor-targeted therapy. As 6 methoxypurine 2'-deoxyriboside (MoPdR) is similar to 6-methylpurine 2' deoxyriboside in structure, we aimed to evaluate the antitumoral effect of the Escherichia coli purine nucleoside phosphorylase (ePNP) gene, using an attenuated salmonella-mediated delivery system, in combination with MoPdR. A novel mutant serovar Typhimurium (SC36) was used to carry the pEGFP-C1-ePNP vector that contains an enhanced green fluorescent protein and an ePNP gene under the control of the cytomegalovirus promoter. The function of the ePNP expression vector was confirmed in vitro using the enzymic conversion of MoPdR into methoxypurine. We also observed a high bystander effect induced by the ePNP/MoPdR system with a very low proportion (1%) of ePNP-positive cells and 5 microg/mL MoPdR, although the growth of parental cells was affected appreciably by MoPdR. The killing effect and increased apoptosis induced by SC36 carrying the ePNP expression vector (SC/ePNP) were detected by cytotoxicity assay and propidium iodide staining flow cytometry analysis, in combination with MoPdR. SC/ePNP was given orally to mice bearing mammary carcinomas, and its antitumor effect was evaluated. SC/ePNP plus MoPdR significantly inhibited tumor growth by approximately 86.6-88.7% and prolonged the survival of tumor-hosting mice. Our data support the view that MoPdR combined with the ePNP gene could be used in gene-directed enzyme prodrug therapy. Attenuated salmonella could be a promising strategy to improve ePNP/MoPdR bystander killing due to its preferential accumulation and anticancer activity in tumors. PMID- 18429959 TI - Risk of metachronous squamous cell carcinoma in the upper aerodigestive tract of Japanese alcoholic men with esophageal squamous cell carcinoma: a long-term endoscopic follow-up study. AB - East Asian case-control studies have shown a strong relationship between alcohol consumption combined with inactive heterozygous aldehyde dehydrogenase-2 (ALDH2*1/*2) and the development of squamous cell carcinoma (SCC), especially multiple SCC, of the upper aerodigestive tract (UADT). This study aimed to identify determinants of the development of metachronous SCC in the UADT in alcoholics with esophageal SCC. Follow-up endoscopic examinations were carried out 4-160 months (median, 41 months) after initial diagnosis in 110 Japanese alcoholic men with esophageal SCC diagnosed by screening using endoscopy combined with oropharyngolaryngeal inspection and esophageal iodine staining. ALDH2*1/*2 was significantly associated with the presence of multiple primary intraesophageal SCC at the time of initial diagnosis. Metachronous primary SCC of the esophagus was diagnosed in 29 of the 81 patients whose initial esophageal SCC was treated by endoscopic mucosal resection alone, and metachronous primary SCC of the oropharyngolarynx was diagnosed in 23 of the 99 patients without synchronous primary SCC of the oropharyngolarynx at the time of initial diagnosis. The risks of metachronous esophageal SCC and oropharyngolaryngeal SCC were significantly higher in ALDH2*1/*2 heterozygotes than in ALDH2*1/*1 homozygotes (age-adjusted and alcohol-adjusted hazard ratio = 3.38 [95% confidence interval: 1.45-7.85] and 4.27 [1.42-12.89], respectively), and in patients with multiple intraesophageal SCC at the time of initial diagnosis than in patients with a solitary intraesophageal SCC (3.09 [1.41-6.78] and 3.25 [1.41 7.47], respectively). ALDH2*1/*2 and multiple synchronous intraesophageal SCC were found to be predictors of metachronous SCC in the UADT in this population. PMID- 18429960 TI - Enhanced distribution of NK012, a polymeric micelle-encapsulated SN-38, and sustained release of SN-38 within tumors can beat a hypovascular tumor. AB - Human pancreatic cancer is generally hypovascular in nature and rich in interstitium. These pathological barriers may contribute to the intractable nature of pancreatic cancer by binding the penetration of anticancer agents throughout the tumor tissue. The aim of the present study was to determine whether NK012 is an appropriate formulation for the treatment of hypovascular tumors. Among pancreatic tumor xenografts, PSN1 appeared to have the richest tumor vasculature and the least number of stromal cells and matrix. In contrast, Capan1 had the poorest tumor vasculature and most abundant stromal tissue. Fluorescence microscopy and high-performance liquid chromatography analysis demonstrated that although NK012 accumulated and continued to be distributed for more than 48 h throughout the entire body of both tumors, CPT-11 disappeared almost entirely from both tumors within 6 h. In addition, efficient sustained release of SN-38 was maintained for more than 96 h in both tumors following administration of NK012. Following the administration of CPT-11, SN-38 was no longer detectable after 24 h in the Capan1 tumor or after 48 h in the PSN1 tumor. All tumors were eradicated in the mice treated with NK012 but not in those treated with CPT-11. Because the antitumor activity of SN-38 is time dependent, NK012, which combines enhanced distribution with sustained release of SN-38 within tumors, may be ideal for the treatment of hypovascular tumors, such as pancreatic cancer. PMID- 18429961 TI - Phase 1 trial of denosumab safety, pharmacokinetics, and pharmacodynamics in Japanese women with breast cancer-related bone metastases. AB - Denosumab, a fully human monoclonal antibody to receptor activator of nuclear factor-kappa B ligand (RANKL), suppresses bone resorption. This open-label, multicenter, phase 1 study evaluated the safety, pharmacodynamics, and pharmacokinetics of denosumab in Japanese women with breast cancer-related bone metastases. Patients (n = 18; median age, 57 years) received a single subcutaneous injection of denosumab 60 mg or 180 mg or three doses of denosumab 180 mg on days 1, 29, and 57 (every 4 weeks) and were followed for > or = 141 days. No major safety concerns related to denosumab were noted in any cohort. All patients experienced at least 1 adverse event (AE); most were mild (grade < or = 2). One patient reported grade 4 myositis and grade 3 anemia, malaise, and dysphagia that the investigator deemed treatment-related; other treatment-related AE were grade < or = 2. No antidenosumab antibodies or clinically significant changes in laboratory findings, vital signs, or electrocardiograms were observed. Pharmacokinetics were approximately dose-linear. Denosumab caused rapid, substantial, and sustained suppression of urinary N-telopeptide corrected for creatinine (uNTx/Cr) across all doses; at day 85, the median change from baseline uNTx/Cr ranged from -61.9% to -90.8%. No dose-limiting toxicity was observed at any dosage. Coupled with pharmacokinetic and pharmacodynamic data, these results were consistent with those observed in non-Japanese populations. PMID- 18429962 TI - Oncogenic role of miR-17-92 cluster in anaplastic thyroid cancer cells. AB - Micro RNAs (miRNAs) are non-coding small RNAs and constitute a novel class of negative gene regulators that are found in both plants and animals. Several miRNAs play crucial roles in cancer cell growth. To identify miRNAs specifically deregulated in anaplastic thyroid cancer (ATC) cells, we performed a comprehensive analysis of miRNA expressions in ARO cells and primary thyrocytes using miRNA microarrays. MiRNAs in a miR-17-92 cluster were overexpressed in ARO cells. We confirmed the overexpression of those miRNAs by Northern blot analysis in ARO and FRO cells. In 3 of 6 clinical ATC samples, miR-17-3p and miR-17-5p were robustly overexpressed in cancer lesions compared to adjacent normal tissue. To investigate the functional role of these miRNAs in ATC cells, ARO and FRO cells were transfected with miRNA inhibitors, antisense oligonucleotides containing locked nucleic acids. Suppression of miR-17-3p caused complete growth arrest, presumably due to caspase activation resulting in apoptosis. MiR-17-5p or miR-19a inhibitor also induced strong growth reduction, but only miR-17-5p inhibitor led to cellular senescence. On the other hand, miR-18a inhibitor only moderately attenuated the cell growth. Thus, we have clarified functional differences among the members of the cluster in ATC cells. In conclusion, these findings suggest that the miR-17-92 cluster plays an important role in certain types of ATCs and could be a novel target for ATC treatment. PMID- 18429963 TI - Valproic acid resensitizes cisplatin-resistant ovarian cancer cells. AB - Although certain inhibitors of histone deacetylases have been shown to induce cytotoxicity alone or in combination with chemotherapeutic agents in cancer cells, the molecular mechanism is not clear. The goal of the present study was to determine whether the antiseizure drug valproic acid (2-propylpentanoic acid; VPA), which is also able to inhibit histone deacetylase, exhibits synergistic cytotoxicity with cisplatin, and the possible pathways for this. Our results clearly show that VPA not only exhibits synergistic cytotoxicity with cisplatin in all of the ovarian carcinoma cells tested, but also can resensitize the cells that have acquired resistance to cisplatin. Consistent with the increased cytotoxicity, cotreatment with VPA was shown to upregulate the cisplatin-mediated DNA damage revealed by phosphorylation of ataxia telangiectasia mutation and histone H2AX. Reactive oxygen species accumulation and tumor suppressor phosphatase and tensin homolog (PTEN) overexpression, which could contribute to the enhanced cytotoxicity, were also observed to be upregulated by VPA. Because PTEN knockdown by small interference RNA or antioxidant treatment can reduce cisplatin-mediated cytotoxicity, it is suggested that upregulation of PTEN and reactive oxygen species by VPA contributes to the enhancement of cisplatin mediated cytotoxicity. These results with resensitization of cisplatin-resistant cells particularly may provide benefits in the treatment of ovarian cancer patients. PMID- 18429964 TI - Communally breeding Bechstein's bats have a stable social system that is independent from the postglacial history and location of the populations. AB - Investigating macro-geographical genetic structures of animal populations is crucial to reconstruct population histories and to identify significant units for conservation. This approach may also provide information about the intraspecific flexibility of social systems. We investigated the history and current structure of a large number of populations in the communally breeding Bechstein's bat (Myotis bechsteinii). Our aim was to understand which factors shape the species' social system over a large ecological and geographical range. Using sequence data from one coding and one noncoding mitochondrial DNA region, we identified the Balkan Peninsula as the main and probably only glacial refugium of the species in Europe. Sequence data also suggest the presence of a cryptic taxon in the Caucasus and Anatolia. In a second step, we used seven autosomal and two mitochondrial microsatellite loci to compare population structures inside and outside of the Balkan glacial refugium. Central European and Balkan populations both were more strongly differentiated for mitochondrial DNA than for nuclear DNA, had higher genetic diversities and lower levels of relatedness at swarming (mating) sites than in maternity (breeding) colonies, and showed more differentiation between colonies than between swarming sites. All these suggest that populations are shaped by strong female philopatry, male dispersal, and outbreeding throughout their European range. We conclude that Bechstein's bats have a stable social system that is independent from the postglacial history and location of the populations. Our findings have implications for the understanding of the benefits of sociality in female Bechstein's bats and for the conservation of this endangered species. PMID- 18429965 TI - High population differentiation and unusual haplotype structure in a shade intolerant pioneer tree species, Zanthoxylum ailanthoides (Rutaceae) revealed by analysis of DNA polymorphism at four nuclear loci. AB - Differences in demographic history, life-history traits, and breeding systems affect nucleotide variation patterns. It is expected that shade-intolerant pioneer tree species have different patterns of genetic polymorphism and population structure than climax species. We studied patterns of nucleotide polymorphism at four putative starch pathway loci (agpSA, agpSB, agpL, and GBSSI) in Zanthoxylum ailanthoides, a shade-intolerant pioneer tree species that occupies forest gaps in warm-temperate forests of East Asia. Genetic diversity was lower within each population than among populations, and differentiation among populations was high across the loci (F(ST) = 0.32-0.64), as expected from the insect-pollinated breeding system and the metapopulation structure of this pioneer species. Numbers of haplotypes were smaller than those expected from the observed numbers of segregating sites. Single haplotypes accounted for more than 47% of all the sampled genes at the respective loci. These variation patterns were incompatible with neutral predictions for populations of a finite island model. Complex population dynamics, such as bottleneck and/or admixture, in the history of this pioneer tree species might have resulted in the observed patterns of genetic variation and population structure, which are different from those of climax wind-pollinated tree species, such as conifers. In contrast to the other loci investigated in this study, agpL showed nearly no variation in Z. ailanthoides (one singleton only), but there was some extent of variation in a closely related species, Zanthoxylum schinifolium. This suggests possibly a recent selective sweep at or near the locus in Z. ailanthoides. PMID- 18429966 TI - Transthyretin accelerates vascular Abeta deposition in a mouse model of Alzheimer's disease. AB - Transthyretin (TTR) binds amyloid-beta (Abeta) and prevents Abeta fibril formation in vitro. It was reported that the lack of neurodegeneration in a transgenic mouse model of Alzheimer's disease (AD) (Tg2576 mouse) was associated with increased TTR level in the hippocampus, and that chronic infusion of anti TTR antibody into the hippocampus of Tg2576 mice led to increased local Abeta deposits, tau hyperphosphorylation and apoptosis. TTR is, therefore, speculated to prevent Abeta pathology in AD. However, a role for TTR in Abeta deposition is not yet known. To investigate the relationship between TTR and Abeta deposition, we generated a mouse line carrying a null mutation at the endogenous TTR locus and the human mutant amyloid precursor protein cDNA responsible for familial AD (Tg2576/TTR(-/-) mouse) by crossing Tg2576 mice with TTR-deficient mice. We asked whether Abeta deposition was accelerated in Tg2576/TTR(-/-) mice relative to the heterozygous mutant Tg2576 (Tg2576/TTR(+/-)) mice. Contrary to our expectations, the degree of total and vascular Abeta burdens in the aged Tg2576/TTR(-/-) mice was significantly reduced relative to the age-matched Tg2576/TTR(+/-) mice. Our experiments present, for the first time, compelling evidence that TTR does not suppress but rather accelerates vascular Abeta deposition in the mouse model of AD. PMID- 18429967 TI - Lack of tacrolimus circadian pharmacokinetics and CYP3A5 pharmacogenetics in the early and maintenance stages in Japanese renal transplant recipients. AB - AIMS: We investigated whether tacrolimus pharmacokinetics shows circadian variation and the influence of the CYP3A5 A6986G polymorphism on the pharmacokinetics in both the early and maintenance stages after renal transplantation. METHODS: Tacrolimus was administered twice daily at specified times (09.00 and 21.00 h) throughout the pre- and post-transplant period according to the trough-targeting strategy. Fifty recipients with stable graft function were studied on day 28 and beyond 1-year post transplantation. Whole blood samples were collected prior to and 1, 2, 3, 4, 6, 9 and 12 h after both the morning and evening doses during hospitalization. RESULTS: Tacrolimus pharmacokinetics did not show circadian variation in either the early or maintenance stage [AUC(0-12) 197.1 (95% confidence interval 182.9, 212.3) in daytime vs. 203.6 ng h ml(-1) (189.8, 217.4) in the night-time at day 28, 102.0 (92.1, 111.9) vs. 107.7 (97.9, 117.5) at 1 year, respectively]. In CYP3A5 *1 allele carriers (CYP3A5 expressers), body weight-adjusted oral clearance was markedly decreased from the early stage to the maintenance stage [0.622 (0.534, 0.709) to 0.369 l h(-1) kg(-1) (0.314, 0425)] compared with a smaller decrease [0.368 (0.305, 0.430) to 0.305 (0.217, 0.393)] in CYP3A5 non-expressers; however, the CYP3A5 genetic variation did not influence tacrolimus chronopharmacokinetics. CONCLUSION: Equivalent daytime and night-time tacrolimus pharmacokinetics were achieved during both the early and maintenance stages with our specified-time administration strategy. The CYP3A5 polymorphism may be associated with the time dependent changes in the oral clearance of tacrolimus, suggesting that genotyping of this polymorphism is useful for determining the appropriate dose of tacrolimus in both the early and maintenance stages after renal transplantation. PMID- 18429968 TI - The ABCG2 C421A polymorphism does not affect oral nitrofurantoin pharmacokinetics in healthy Chinese male subjects. AB - AIMS: A number of drugs are substrates or inhibitors of the efflux transporter breast cancer resistance protein (BCRP; ABCG2), which can limit systemic exposure by reducing absorption and/or increasing biliary elimination. The identification of a BCRP-selective clinical probe drug would provide a useful tool to understand the effect of genetic polymorphisms and transporter-based drug interactions on drug pharmacokinetics. The aim of this study was to assess the utility of nitrofurantoin as a clinical probe substrate for BCRP activity by evaluating the impact of genetic variation on nitrofurantoin pharmacokinetics. METHODS: Nitrofurantoin pharmacokinetics were studied in an open-label, single-oral dose (100 mg) study in 36 male Chinese subjects who were pre-screened for ABCG2 421 CC, CA and AA genotypes (n = 12 each). Plasma and urine concentrations of nitrofurantoin were determined by LC/MS/MS and LC/UV respectively. anova was used to compare pharmacokinetic parameters among genotypes. RESULTS: There were no significant differences in nitrofurantoin pharmacokinetics among the genotypic cohorts. The geometric mean nitrofurantoin plasma AUC((0-infinity)) (95% confidence interval) values were 2.21 (2.00, 2.45), 2.42 (2.11, 2.78) and 2.32 (1.99, 2.70) microg h ml(-1) and half-life values were 0.79 (0.59, 1.0), 0.76 (0.64, 0.89) and 0.72 (0.62, 0.84) h for ABCG2 421 genotypes CC, CA and AA, respectively. The percentage of dose excreted unchanged in the urine was 43, 44 and 39%, respectively. CONCLUSIONS: The ABCG2 C421A polymorphism had no effect on nitrofurantoin plasma and urine pharmacokinetic parameters in healthy Chinese subjects. These results indicate that nitrofurantoin is not a suitable clinical probe substrate for assessing BCRP activity. PMID- 18429969 TI - Pharmacogenetics of aspirin resistance: a comprehensive systematic review. AB - AIMS: The aim was to perform a systematic review of all candidate gene association studies in aspirin resistance. METHODS: Electronic databases were searched up until 1 December 2007 for all studies investigating any candidate gene for aspirin resistance in humans. Aspirin resistance was required to have been measured by a standardized laboratory technique to be included in the analysis. RESULTS: Within 31 studies, 50 polymorphisms in 11 genes were investigated in 2834 subjects. The PlA1/A2 polymorphism in the GPIIIa platelet receptor was the most frequently investigated, with 19 studies in 1389 subjects. The PlA1/A2 variant was significantly associated with aspirin resistance when measured in healthy subjects [odds ratio (OR) 2.36, 95% confidence interval (CI) 1.24, 4.49; P = 0.009]. Combining genetic data from all studies (comprising both healthy subjects and those with cardiovascular disease) reduced the observed effect size (OR 1.14, 95% CI 0.84, 1.54; P = 0.40). Moreover, the observed effect of PlA1/A2 genotype varied depending on the methodology used for determining aspirin sensitivity/resistance. No significant association was found with aspirin resistance in four other investigated polymorphisms in the COX-1, GPla, P2Y1 or P2Y12 genes. CONCLUSIONS: Our data support a genetic association between the PlA1/A2 molecular variant and aspirin resistance in healthy subjects, with the effect diminishing in the presence of cardiovascular disease. The laboratory methodology used influences the detection of aspirin resistance. However, as heterogeneity was significant and our results are based on a limited number of studies, further studies are required to confirm our findings. PMID- 18429970 TI - The association of adiponectin allele 45T/G and -11377C/G polymorphisms with Type 2 diabetes and rosiglitazone response in Chinese patients. AB - WHAT IS ALREADY KNOWN ABOUT THIS SUBJECT: Rosiglitazone is able to increase serum adiponectin levels significantly in Type 2 diabetic patients. :The role of genetic factors that determine the marked interindividual variability in glucose lowering efficacy of rosiglitazone in Chinese patients is not known. The current study was designed to evaluate the impact of the adiponectin common allele 45T/G and -11377C/G polymorphisms on the response to rosiglitazone monotherapy in Chinese patients with Type 2 diabetes (T2D). WHAT THIS STUDY ADDS: The genetic polymorphisms of adiponectin alleles 45T/G and -11377C/G as well as their common diplotypes are significantly associated with an attenuated fasting plasma glucose, postprandial plasma glucose and homeostasis model assessment for insulin resistance as well as an enhanced adiponectin concentration in Chinese patients with T2D after rosiglitazone treatment. AIMS The aim of the present study was to evaluate the impact of adiponectin allele T45G and C-11377G genetic polymorphisms on efficacy of rosiglitazone in Chinese patients with type 2 diabetes (T2D). METHODS: Patients with T2D (n = 255) and 120 healthy volunteers were enrolled to identify 45T/G and -11377C/G genotypes by polymerase chain reaction-restriction fragment length polymorphism assay. Forty-two T2D patients with different 45T/G or -11377C/G genotypes received orally rosiglitazone as a single-dose therapy (4 mg day-1 p.o.) for 12 weeks. Serum triglyceride, fasting plasma glucose (FPG), postprandial plasma glucose (PPG), glycated hemoglobin, fasting serum insulin, postprandial serum insulin, total cholesterol, homeostasis model assessment for insulin resistance (HOMA-IR), low-density lipoprotein-cholesterol, high-density lipoprotein-cholesterol (HDL-c) and adiponectin concentration were determined before and after rosiglitazone treatment. RESULTS: We showed an attenuated rosiglitazone effect in patients with -11377CG+GG heterozygote genotype on FPG, PPG, HOMA-IR compared with -11377CC homozygote genotype. However, we found an enhanced rosiglitazone effect on serum adiponectin concentration in patients with -11377CC homozygote genotype compared with -11377CG+GG heterozygote genotype (P = 0.000) and in patients with 45TG + GG heterozygote genotype compared with 45TT homozygote genotype (P = 0.018). Finally, our results showed that there was an enhanced effect in patients with -11377/45 CGTT diplotype compared with other discovered diplotypes on FPG (P = 0.001) and PPG (P = 0.003) after rosiglitazone treatment. CONCLUSIONS: These data suggest that the adiponectin allele 45T/G and 11377C/G polymorphisms are significantly associated with the therapeutic efficacy of multiple-dose rosiglitazone in Chinese patients with T2D. PMID- 18429971 TI - Matrix metalloproteinase-9 and metalloproteinase-2 activity and expression is reduced by melatonin during experimental colitis. AB - Matrix metalloproteinases (MMPs) are associated with matrix turnover in both physiological and pathological conditions. Several data indicate that MMPs play an important role in the pathogenesis of colitis. Various evidence has documented that the pineal secretory product melatonin exerts an important anti-inflammatory effect in different experimental models including colitis. However, no reports are available on the relationship between the activity and expression of MMPs and anti-inflammatory effect of melatonin. The aim of the present study was to evaluate whether melatonin prevents the experimental colitis in rats by regulating MMP-9 and MMP-2 activity and expression. Colitis was induced by intracolonic instillation of dinitrobenzene sulphonic acid (DNBS). Four days after DNBS administration, colon TNF-alpha production was associated with colon damage. Biochemical methods and zymography were used to analyse MMP-9 and MMP-2 activities in colon tissues from DNBS-injured rats. Our studies reveal that melatonin prevented colon injury and lipid peroxidation in rats at 4 days after DNBS-induced colitis. Melatonin also reduced proMMP-9 and MMP-2 activities that were induced in the colon tissues by DNBS administration. Reduced MMP-9 and MMP-2 activities were associated with reduced expression of TNF-alpha. We conclude that melatonin's ability to reduce DNBS-induced colon injury in rats is related to a reduction in proMMP-9 and MMP-2 activities and expression. PMID- 18429972 TI - The effects of a maximal jaw clench on an inhibitory jaw reflex. AB - The purpose of this study was to determine whether mimicking symptoms of temporomandibular disorders by experimentally activating deep nociceptors in the oro-facial region, can modulate an inhibitory jaw reflex. In human subjects, electromyograms were recorded from one (eight subjects) or both (16 subjects) active masseter muscles and electrical stimuli were applied to the upper lip. This procedure was performed before and after a 30-s conditioning period in which the subjects maximally clenched the jaw. In all subjects, the electrical stimuli produced an inhibition of masseter activity. Following conditioning, there was a small but not statistically significant decrease in the mean size of this inhibition (anova: P = 0.066 and P = 0.077, for responses recorded ipsi- and contralaterally to the stimulus). There was no relationship between changes in the reflex and the levels of pain induced by the conditioning procedure and recorded on 100 mm visual analogue scales (range = 0-64 mm, median = 11 mm) (Spearman's correlation test: P = 0.412). These findings suggest that inhibitory jaw reflexes are little if at all affected by this conditioning procedure. The hypothesis that temporomandibular disorders both cause and are sustained by a decrease in protective jaw reflexes is not supported by these findings. PMID- 18429973 TI - The use of CT scan based planning for oral rehabilitation by means of implants and its transfer to the surgical field: a critical review on accuracy. AB - The purpose of the present paper is to review the literature on the use of CT scan based planning for oral rehabilitation and its transfer to the surgical field by means of a surgical guide. The first part will deal with surgical guides based on tomographies or CT scan data often using dedicated software, but manually fabricated. In the second part, CT-derived drill guides are discussed, which are fabricated by means of CAD/CAM technology or other computer-controlled technology. The deviations between the position of the implants at the planning stage and after the surgery are of utmost importance, especially when flapless procedures are applied. The maximal deviations are often not stated in the literature. This should be taken into account when a system is applied clinically. PMID- 18429974 TI - Collagenous microstructure of the glenoid labrum and biceps anchor. AB - The glenoid labrum is a significant passive stabilizer of the shoulder joint. However, its microstructural form remains largely unappreciated, particularly in the context of its variety of functions. The focus of labral microscopy has often been histology and, as such, there is very little appreciation of collagen composition and arrangement of the labrum, and hence the micromechanics of the structure. On transmission electron microscopy, significant differences in diameter, area and perimeter were noted in the two gross histological groups of collagen fibril visualized; this suggests a heterogeneous collagenous composition with potentially distinct mechanical function. Scanning electron microscopy demonstrated three distinct zones of interest: a superficial mesh, a dense circumferential braided core potentially able to accommodate hoop stresses, and a loosely packed peri-core zone. Confocal microscopy revealed an articular surface fine fibrillar mesh potentially able to reduce surface friction, bundles of circumferential encapsulated fibres in the bulk of the tissue, and bone anchoring fibres at the osseous interface. Varying microstructure throughout the depth of the labrum suggests a role in accommodating different types of loading. An understanding of the labral microstructure can lead to development of hypotheses based upon an appreciation of this component of material property. This may aid an educated approach to surgical timing and repair. PMID- 18429975 TI - Definition of the capsular insertion plane on the proximal humerus. AB - The aim of this work was quantitatively to establish the relationship between the plane that hosts the humeral head lateral margin (anatomical neck) and that of the capsular insertion. Eight cadaveric shoulders were used. These were dissected, exposing the humeral head margin and the root of the capsular humeral insertion to extract digitally their outlines using a mechanical 3-d digitizer. The datasets of the digitized outlines were applied and the geometric planes they best fitted mathematically calculated. Vector analysis techniques were finally applied to the two planes to quantify the relationship between them. The humeral head margin is circular (+/- 2.2% of radius), having each of its outlining points on the same plane (within +/- 1.5 mm.) The capsular attachment outlining points also insert on a plane (+/- 1.4 mm). The two planes are related to one another by an inclination of 14.5 +/- 3.6 degrees. The relationship described here would allow for in vivo prediction of humeral attachment of capsular structures by using radiological datasets of the anatomical neck. This would be useful in patient-specific modelling to study and understand the glenohumeral ligament kinematics during clinical examinations and to plan surgical reconstructive procedures. PMID- 18429976 TI - Pulsed electric field treatment as a potential method for microbial inactivation in scaffold materials for tissue engineering: the inactivation of bacteria in collagen gel. AB - AIMS: To investigate the effectiveness of pulsed electric field (PEF) treatment as a new method for inactivation of micro-organisms in complex biomatrices and to assess this by quantifying the inactivation of Escherichia coli seeded in collagen gels. METHODS AND RESULTS: PEF was applied to E. coli seeded collagen gels in static (nonflowing) chambers. The influence of electric field strength, pulse number and seeded cell densities were investigated. The highest level of inactivation was obtained at the maximum field strength of 45 kV cm(-1). For low levels of E. coli contamination (10(3) CFU ml(-1)), PEF treatment resulted in no viable E. coli being recovered from the gels. However, PEF treatment of gels containing higher cell densities (>or=10(4) CFU ml(-1)) did not achieve complete inactivation of E. coli. CONCLUSIONS: PEF treatment successfully inactivated E. coli seeded in collagen gels by 3 log(10) CFU ml(-1). Complete inactivation was hindered at high cell densities by the tailing effect observed. SIGNIFICANCE AND IMPACT OF THE STUDY: PEF shows potential as a novel, nondestructive method for decontamination of collagen-based matrices. Further investigation is required to ensure its compatibility with other proteins and therapeutic drugs for tissue engineering and drug delivery applications. PMID- 18429977 TI - Water potential changes in faecal matter and Escherichia coli survival. AB - AIMS: This study investigated the influence of a range of evaporation rates (2.0, 5.3 and 7.4 mm day(-1)) on degradation of E. coli (ATCC Strain 25922) inoculated in canine faeces. METHODS AND RESULTS: Experiments were carried out in an environmental chamber and a first order exponential decay function (Chick's Law) was used to estimate degradation rates. We estimated die-off coefficients using linear regression. Die-off rates were -0.07, -0.22 and -0.23 h(-1), respectively, for evaporation rates of 2.0, 5.3 and 7.4 mm day(-1) (P = 0.000+, for each model). Nearly complete die-off was found within 15-60 h (7.4-2.0 mm day(-1) evaporation rates), which corresponds with a water potential of approximately 22.4 MPa. CONCLUSIONS: This study indicates that canine faeces need not be desiccated to achieve complete loss of indicator organisms. Water potential, which is a combination of osmotic and matric potential, is a key stress that increases as evaporation removes water from the faecal matrix and increases concentration of the remaining faecal solution. Evaporation may remove populations of indicator organisms in faeces relatively quickly, even though faeces are not completely dehydrated. SIGNIFICANCE AND IMPACT OF THE STUDY: This research may be used as the foundation for studies more closely resembling real world evaporation conditions including diurnal fluctuations, rewetting and freezing. PMID- 18429978 TI - Microbial DNA profiling by multiplex terminal restriction fragment length polymorphism for forensic comparison of soil and the influence of sample condition. AB - AIMS: To evaluate: (i) the impact of air-drying on bacterial, archaeal and fungal soil DNA profiles and (ii) the potential use of multiplex-terminal restriction fragment length polymorphism (M-TRFLP) as a tool for forensic comparison of soil. METHODS AND RESULTS: An M-TRFLP approach was used to profile bacterial, archaeal and fungal DNA profiles from five different soil sites. Air-drying soil significantly reduced the quantity of DNA but the number of operational taxanomic units (OTU) was unaffected. The impact of air-drying on soil DNA profiles was dependent on soil site and microbial primers. Fungal profiles were altered the least by air-drying. For prokaryotic profiles, air-drying altered the relative similarity/dissimilarity between soil sites. The M-TRFLP approach was more discriminatory compared with soil colour and single-taxa profiling, but did not significantly improve resolution between two similar soils. CONCLUSIONS: Of those tested, soil fungi were potentially the more robust target for application to soil forensic studies as they were altered less by air-drying and provided clear discrimination of soils from different sites. The M-TRFLP method demonstrated potential to achieve greater resolution, discriminating the soil sites based on both bacterial and fungal components. SIGNIFICANCE AND IMPACT OF THE STUDY: Soil DNA profiling has potential as a forensic tool, but sample condition and the appropriate selection of microbial target taxa must be considered. PMID- 18429979 TI - Healing of sickle cell ulcers during pregnancy: a favourable effect of foetal cell transfer? PMID- 18429980 TI - Occupational contact dermatitis due to polyvinylamine. PMID- 18429981 TI - Chronic nail biting and irreversible shortening of the fingernails. PMID- 18429982 TI - Epidermal pseudocarcinomatous hyperplasia with underlying epidermal growth factor producing cutaneous CD30-positive lymphoproliferative disorder. PMID- 18429983 TI - Disseminated mucosa-associated lymphoid tissue lymphoma involving the skin and multiple mucosal sites. PMID- 18429984 TI - Photodynamic therapy in disseminated superficial actinic porokeratosis. PMID- 18429985 TI - Identification of two recurrent mutations in keratin genes in three cases with pachyonychia congenita. PMID- 18429986 TI - Case report on Malassezia infection of palms and fingernails--speculations on cause for therapeutic failure in pityriasis versicolor. PMID- 18429987 TI - Molluscum contagiosum of the sole. PMID- 18429988 TI - Adult-onset Still's disease: an uncommon differential diagnosis of urticaria and treatment with anakinra. PMID- 18429989 TI - Pulsed itraconazole therapy in eruptive lichen planus. PMID- 18429990 TI - Endothelial nitric oxide synthase is a critical factor in experimental liver fibrosis. AB - Reduced expression of endothelial nitric oxide synthase (eNOS) in chronic liver disease can reduce hepatic perfusion and accelerate fibrosis. The relationship between eNOS expression and liver fibrogenesis remains unclear. We investigated whether L-arginine attenuated chronic liver fibrosis through eNOS expression. Chronic liver injury was induced by administration of carbon tetrachloride (CCl(4)) to mice for 8 weeks. 5-Methylisothiourea hemisulphate (SMT), an iNOS inhibitor, or L-arginine, a NOS substrate were injected subcutaneously. CCl(4) induced hepatotoxicity, oxidative stress and accumulation of collagen were detected in the liver. The expression levels of inducible NOS (iNOS) and nuclear factor kappa-B (NF-kappaB) activity in the liver after CCl(4) treatment were increased but eNOS expression and activator protein-1 (AP-1) activity were decreased. Both SMT and L-arginine effectively reduced CCl(4) induced oxidative stress and collagen formation, but L-arginine showed a significantly greater suppression of collagen formation, iNOS expression and NF-kappaB activity. L arginine also restored the level of eNOS and AP-1 activity. L-arginine was more effective than SMT in suppressing liver fibrosis. L-arginine might improve NO production which facilitates hepatic blood flow and thus retards liver fibrogenesis. Our results showed that the reduced eNOS expression in CCl(4) treated mice was reversed by L-arginine. Furthermore, L-arginine also reversed the reduced AP-1 activity, an eNOS promoter. PMID- 18429992 TI - Listening to children: gaining a perspective of the experiences of poverty and social exclusion from children and young people of single-parent families. AB - This paper reports on the experiences and views of children and young people of single-parent families, as findings from a European Union-funded research project undertaken in England, Greece and Cyprus. The objectives of the research project were to investigate how children and young people of single-parent families understand and experience their worlds as members of these families: whether and to what extent they experience poverty and social exclusion and how they cope with the challenges that this confronts them with. Methodology was replicated in each of the countries; however, this paper draws on the English experiences. Semistructured interviews (40) and focus groups (four) were undertaken with children of single parents. In addition, focus groups were undertaken with children of two-parent families (four), focus groups with single parents, focus groups with two-parent families (four) and individual interviews with key professionals. Detailed discussion guides were followed, with open-ended questions to allow participants to express their feelings and ideas in their own words. The research sample included children from single-parent and two-parent families, aged 6 years to 16 years, balanced in terms of age, gender and geographical location. Findings demonstrate the children's and young people's understanding of the impact of poverty and social exclusion on their family life and their everyday experiences. The positive benefits of being in a single-parent family are highlighted, with 'time poverty' raised as a significant issue. Children and young people are aware of their poverty and how it influences exclusion from friendships, play, leisure and community activities. Policy needs to take account of the systematic reality of children's experiences; alliances with adults that support meaningful involvement and participation by children and young people will make a significant contribution to this. PMID- 18429991 TI - Increased pancreatic islet mass is accompanied by activation of the insulin receptor substrate-2/serine-threonine kinase pathway and augmented cyclin D2 protein levels in insulin-resistant rats. AB - It is well known that glucocorticoids induce peripheral insulin resistance in rodents and humans. Here, we investigated the structural and ultrastructural modifications, as well as the proteins involved in beta-cell function and proliferation, in islets from insulin-resistant rats. Adult male Wistar rats were made insulin resistant by daily administration of dexamethasone (DEX; 1mg/kg, i.p.) for five consecutive days, whilst control (CTL) rats received saline alone. Structure analyses showed a marked hypertrophy of DEX islets with an increase of 1.7-fold in islet mass and of 1.6-fold in islet density compared with CTL islets (P < 0.05). Ultrastructural evaluation of islets revealed an increased amount of secreting organelles, such as endoplasmic reticulum and Golgi apparatus in DEX islets. Mitotic figures were observed in DEX islets at structural and ultrastructural levels. Beta-cell proliferation, evaluated at the immunohistochemical level using anti-PCNA (proliferating cell nuclear antigen), showed an increase in pancreatic beta-cell proliferation of 6.4-fold in DEX islets compared with CTL islets (P < 0.0001). Increases in insulin receptor substrate-2 (IRS-2), phosphorylated-serine-threonine kinase AKT (p-AKT), cyclin D(2) and a decrease in retinoblastoma protein (pRb) levels were observed in DEX islets compared with CTL islets (P < 0.05). Therefore, during the development of insulin resistance, the endocrine pancreas adapts itself increasing beta-cell mass and proliferation, resulting in an amelioration of the functions. The potential mechanisms that underlie these events involve the activation of the IRS 2/AKT pathway and activation of the cell cycle, mediated by cyclin D(2). These adaptations permit the maintenance of glycaemia at near-physiological ranges. PMID- 18429993 TI - Growing up with a lesbian or gay parent: young people's perspectives. AB - Drawing on life story approaches, this paper reports on a qualitative content analysis of 67 published accounts in the UK, USA and New Zealand of young people and adults reflecting on their experiences of growing up with one or more lesbian or gay parent(s). Responses of the young people to their parent's sexuality were categorised as predominantly positive, neutral, ambivalent and somewhat negative. Young people reported that they had experienced homophobic attitudes and behaviours in one or more of three domains: general and institutional, within the family, and from peers/friends and other parents. The implications of the analysis for social care and social work practice in the UK context with this group of children and their families are discussed. PMID- 18429994 TI - Italy and Austria before and after study: second-hand smoke exposure in hospitality premises before and after 2 years from the introduction of the Italian smoking ban. AB - The aim of this paper was to compare nicotine concentration in 28 hospitality premises (HPs) in Florence and Belluno, Italy, where a smoking ban was introduced in 2005, and in 19 HPs in Vienna, Austria, where no anti-smoking law entered into force up to now. Airborne nicotine concentrations were measured in the same HPs in winter 2002 or 2004 (pre-ban measurements) and winter 2007 (post-ban measurements). In Florence and Belluno, medians decreased significantly (P < 0.001) from 8.86 [interquartile range (IQR): 2.41-45.07)] before the ban to 0.01 microg/m3 (IQR: 0.01-0.41) afterwards. In Austria (no smoking ban) the medians collected in winters 2004 and 2007 were, respectively, 11.00 (IQR: 2.53-30.38) and 15.76 microg/m3 (IQR: 2.22-31.93), with no significant differences. Measurements collected in winter 2007 in 28 HPs located in Naples, Turin, Milan (0.01 microg/m3; IQR: 0.01-0.16) confirmed post-ban results in Florence and Belluno. The medians of nicotine concentrations in Italy and Austria before the Italian ban translates, using the risk model of Repace and Lowery, into a lifetime excess lung cancer mortality risk for hospitality workers of 11.81 and 14.67 per 10,000, respectively. Lifetime excess lung cancer mortality risks for bar and disco-pub workers were 10-20 times higher than that calculated for restaurant workers, both in Italy and Austria. In winter 2007, it dropped to 0.01 per 10,000 in Italy, whereas in Austria it remained at the same levels. The drop of second-hand smoke exposure indicates a substantial improvement in air quality in Italian HPs even after 2 years from the ban. PRACTICAL IMPLICATIONS: The nation-wide smoking ban introduced in Italy on January 10, 2005, resulted in a drop in second-hand smoke exposure in hospitality premises, whereas in Austria, where there is no similar nation-wide smoking ban, the exposure to second-hand smoke in hospitality premises remains high. Given that second-hand smoke is considered a group 1 carcinogen according to the International Agency for Research on Cancer classification, the World Health Organization Framework Convention on Tobacco Control strongly recommends the implementation of nation wide smoke-free policies in order to improve the indoor air quality of hospitality premises and workplaces. Results from our study strongly supports this recommendation. PMID- 18429995 TI - Monitoring of volatile organic compounds in non-residential indoor environments. AB - A weekly monitoring campaign of volatile organic compounds (VOC), with single sampling of 24 h, was carried out in non-residential indoor environments such as libraries, pharmacies, offices, gymnasiums, etc., in order to evaluate the VOC concentrations to which people are exposed. Moreover, an outdoor sample was coupled to each indoor site to point out the influence of indoor sources. They were sampled with Radiello diffusive samplers for thermal desorption and analyzed by GC-MS. As already described in other papers, the VOC levels of most of the indoor sites were higher than that observed in the corresponding outdoor sites. For example, some sites showed a level of pollution that is ten times higher than their corresponding outdoor site. The monitored environments that had higher concentrations of the investigated VOC were the pharmacies, a newspaper stand, a copy center, and the coffee shops. Analysis of the weekly average concentrations of each pollutant and the use of literature allowed pointing out some site specific characteristics that singled out possible sources of VOC. These results were verified analyzing the indoor-outdoor ratio (I/O) too. Newspaper stands were characterized by very high concentrations of toluene and pharmacies were characterized by high concentrations of aromatic compounds. PRACTICAL IMPLICATIONS: Indoor air pollution caused by volatile organic compounds (VOC) might affect human health at home as well as in public and commercial buildings. The main VOC sources in indoor environments are human activities, personal care products, smoking, house cleaning products, building products, and outside pollution. To preserve human health it is necessary to evaluate the average concentrations of VOC to which people are exposed and to identify the main sources of indoor pollution by means of suitable indoor monitoring campaigns in several environments. These investigations allow pointing out the characteristic critical situations of some indoor environments or some other types of environments. PMID- 18429996 TI - Use of (1-3)-beta-d-glucan concentrations in dust as a surrogate method for estimating specific fungal exposures. AB - Indoor exposure to fungi has been associated with respiratory symptoms,often attributed to their cell wall component, (1-3)-beta-D-glucan. Performing(1-3) beta-D-glucan analysis is less time consuming and labor intensive than cultivation or microscopic counting of fungal spores. This has prompted many to use(1-3)-beta-D-glucan as a surrogate for fungal exposure. The aim of this study was to examine which indoor fungal species are major contributors to the (1-3) beta-D-glucan concentration in field dust samples. We used the quantitative polymerase chain reaction (QPCR) method to analyze 36 indoor fungal species in 297 indoor dust samples. These samples were also simultaneously analyzed for (1 3)-beta-D-glucan concentration using the endpoint chromogenic Limulus Amebocyte lysate assay. Linear regression analysis, followed by factor analysis and structural equation modeling, were utilized in order to identify fungal species that mostly contribute to the (1-3)-beta-D-glucan concentration in field dust samples. The study revealed that Cladosporium and Aspergillus genera, as well as Epicoccum nigrum, Penicillium brevicompactum and Wallemia sebi were the most important contributors to the (1-3)-beta-D-glucan content of these home dust samples. The species that contributed most to the (1-3)-beta-D-glucan concentration were also the most prevalent in indoor environments. However, Alternaria alternata, a common fungal species in indoor dust, did not seem to be a significant source of (1-3)-beta-D-glucan. PRACTICAL IMPLICATIONS: This study revealed that the (1-3)-beta-D-glucan content of different fungal species varies widely. (1-3)-beta-D-glucan inhouse dust from the Greater Cincinnati area may be a good marker for some fungal species of the Cladosporium and Aspergillus genera. In contrast, Alternaria alternata did not contribute much to the (1-3)-beta-D glucan load. Therefore, (1-3)-beta-D-glucan concentration in field samples as a surrogate for total fungal exposure should be used with caution. PMID- 18429997 TI - What should be given a priority - costly medications for relatively few people or inexpensive ones for many? The Health Parliament public consultation initiative in Israel. AB - BACKGROUND: In the past two decades, government and civic organizations have been implementing a wide range of deliberative public consultations on health care related policy. Drawing on these experiences, a public consultation initiative in Israel called the Health Parliament was established. GOALS: To implement a public consultation initiative that will engage members of the public in the discussion of four healthcare policy questions associated with equity in health services and on priorities for determining which medications and treatments should be included in the basket of national health services. METHOD: One hundred thirty-two participants from the general population recruited through a random sample were provided with background materials and met over several months in six regional sites. Dilemma activities were used and consultants were available for questions and clarifications. Participants presented their recommendations in a national assembly to the Minister of Health. OUTCOMES: Across the regional groups the recommendations were mostly compatible, in particular regarding considering the healthcare system's monetary state, even at the expense of equity, but for each policy question minority views were also expressed. A strong emphasis in the recommendations was pragmatism. CONCLUSION: Participants felt the experience was worthwhile; though the actual impact of their recommendations on policy making was indirect, they were willing to participate in future consultations. However, despite enthusiasm the initiative was not continued. Issues raised are whether consultation initiatives must have a direct impact on healthcare policy decisions or can be mainly a venue to involve citizens in the deliberation of healthcare policy issues. PMID- 18429998 TI - Scratching the surface of the rare biosphere with ribosomal sequence tag primers. AB - Increasingly large datasets of 16S rRNA gene sequences reveal new information about the extent of microbial diversity and the surprising extent of the rare biosphere. Currently, many of the largest datasets are represented by short and variable ribosomal sequence tags (RSTs) that are limited in their ability to accurately assign sequences to broad-scale phylogenetic trees. In this study, we selected 30 rare RSTs from existing sequence datasets and designed primers to amplify c. 1400 bases of the 16S rRNA gene to determine whether these sequences were represented by existing databases or if they might reveal new lineages within the Bacteria. Approximately one-third of the RST primers successfully amplified longer portions of these low-abundance 16S rRNA genes in a specific manner. Subsequent phylogenetic analysis demonstrated that most of these sequences were (1) distantly related to existing cultivated microorganisms and (2) closely related to uncultivated clone sequences that were recently deposited in GenBank. The presence of so many recently collected 16S rRNA gene reference sequences in existing databases suggests that progress is being made quickly towards a microbial census, one which has begun scratching the surface of the 'rare biosphere'. PMID- 18429999 TI - Site-directed mutagenesis of the temperature-sensing histidine protein kinase CorS from Pseudomonas syringae. AB - Several plant pathogenic bacteria belonging to the species Pseudomonas syringae produce the phytotoxin coronatine to enhance their virulence. Pseudomonas syringae pv. glycinea PG4180 synthesizes coronatine at the virulence-promoting temperature of 18 degrees C, but not at 28 degrees C, its optimal growth temperature. In contrast, temperature has virtually no effect on coronatine synthesis in P. syringae pv. tomato strain DC3000. A modified two-component system controlling coronatine synthesis and consisting of the histidine protein kinase (HPK), CorS, the response regulator, CorR, and a third essential component, CorP, had been identified previously in both strains. CorS had been identified previously as a potential thermo-sensor. Comparison of the amino acid sequences of the HPKs from the two organisms revealed distinct differences. Site directed mutagenesis of CorS from PG4180 was used to identify amino acyl residues potentially important for temperature signal perception. Point mutations and combinations of these were introduced into corS of PG4180 to generate corS variants with increased similarities to the respective allele from strain DC3000. These mutations resulted in either loss of activity, increase of thermoresponsiveness, or had no effect on CorS activity. Although none of the introduced mutations resulted in a clear conversion of CorS activity from thermo responsive to temperature-independent, amino acyl residues important for temperature-dependent CorS activity and coronatine biosynthesis were identified. PMID- 18430000 TI - Predicting sigma28 promoters in eleven Shewanella genomes. AB - An iterative position-specific score matrix (PSSM)-based approach was used to predict sigma(28) promoters in 11 Shewanella genomes. The Shewanella Correlation Browser was used to distinguish true-positive predictions from false-positive predictions in Shewanella oneidensis MR-1 by generating a sigma(28)-regulated transcriptional network from transcriptional profiling data. This dual-pronged approach identified several genes that have sigma(28) promoters and that may be involved with motility or chemotaxis in Shewanella. PMID- 18430001 TI - Localization and function of ADP ribosylation factor A in Aspergillus nidulans. AB - Filamentous fungi undergo polarized hyphal growth throughout the majority of their life cycle. The Spitzenkorper is a structure unique to filamentous fungi that participates in hyphal growth and is composed largely of vesicles. An important class of proteins involved in vesicle assembly and trafficking are the ADP-ribosylation factors (Arfs). In Saccharomyces cerevisiae, Arf1p and Arf2p are involved in secretion. Aspergillus nidulans ArfA is a homolog of ScArf1p and ScArf2p with 75% of amino acid sequence similarity to each. ArfA::GFP localizes to cellular compartments consistent with Golgi equivalents. An N-terminal myristoylation motif is critical for localization of ArfA. Treatment with Brefeldin A, an inhibitor of Golgi transport, leads to ArfA::GFP diffusing through the cytosol and accumulating into a subcellular compartment further suggesting the ArfA localizes to and functions in the Golgi network. Costaining with FM4-64 revealed that ArfA::GFP likely localized to subcellular compartments participating in exocytosis. We were unable to recover arfA gene disruption strains indicating that the gene is essential in A. nidulans. The overexpression of ArfA protein partially suppresses the polarity defect phenotype of an N myristoyltransferase mutant. Taken together, these results suggest that ArfA participates in hyphal growth through the secretory system. PMID- 18430002 TI - Effect of the HP0159 ORF mutation on the lipopolysaccharide structure and colonizing ability of Helicobacter pylori. AB - The outer core region of Helicobacter pylori lipopolysaccharide of the majority of isolates contains an alpha-1,6-glucan polymer synthesized by the product of the HP0159 ORF. Structural studies carried out on HP0159 lipopolysaccharide mutants by a combination of chemical methods, mass spectrometry and nuclear magnetic resonance spectroscopy confirmed that insertional inactivation of HP0159 gene in H. pylori strains 26695 and SS1 resulted in formation of a truncated lipopolysaccharide molecule characterized by the presence of a terminal dd heptose residue in the side-chain outer core fragment and maintaining an inner core backbone structure compared with the wild-type Lewis antigen-expressing strains. Colonization studies with HP0159 mutants of two mouse-colonizing strains, SS1 and M6, confirmed their inability to successfully colonize the murine stomach. PMID- 18430003 TI - Immunostimulatory activity of Bacillus spores. AB - Bacillus species, typically Bacillus subtilis, are being used as probiotics and mounting evidence indicates that Bacillus species are important for development of a robust gut-associated lymphoid system (GALT). We used a number of gut isolates of Bacillus incorporating three species, B. subtilis, Bacillus licheniformis and Bacillus flexus to evaluate the nature of interaction between spores and the GALT. In mice orally administered with spores, evidence of cell proliferation was determined in the germinal centers of Peyer's patches. Stimulation of antigen-presenting cells and T lymphocytes was also markedly enhanced. Cytokines were shown to be induced in spleens and mesenteric lymph nodes of mice including the proinflammatory cytokines, tumour necrosis factor alpha and IL-6. We also demonstrated that vegetative cells of B. subtilis can stimulate expression of the toll-like receptor (TLR) genes for TLR2 and TLR4. However, we were able to show that spores could not stimulate either and must, by default, interact with another TLR and by this mechanism help activate innate immunity. PMID- 18430004 TI - Vegetation cover of forest, shrub and pasture strongly influences soil bacterial community structure as revealed by 16S rRNA gene T-RFLP analysis. AB - Bacterial community structure is influenced by vegetation, climate and soil chemical properties. To evaluate these influences, terminal restriction fragment length polymorphism (T-RFLP) and cloning of the 16S rRNA gene were used to analyze the soil bacterial communities in different ecosystems in southwestern China. We compared (1) broad-leaved forest, shrub and pastures in a high-plateau region, (2) three broad-leaved forests representing a climate gradient from high plateau temperate to subtropical and tropical regions and (3) the humus and mineral soil layers of forests, shrub lands and pastures with open and restricted grazing activities, having varied soil carbon and nutrient contents. Principal component analysis of the T-RFLP patterns revealed that soil bacterial communities of the three vegetation types were distinct. The broad-leaved forests in different climates clustered together, and relatively minor differences were observed between the soil layers or the grazing regimes. Acidobacteria dominated the broad-leaved forests (comprising 62% of the total clone sequences), but exhibited lower relative abundances in the soils of shrub (31%) and pasture (23%). Betaproteobacteria was another dominant taxa of shrub land (31%), whereas Alpha- (19%) and Gammaproteobacteria (13%) and Bacteriodetes (16%) were major components of pasture. Vegetation exerted more pronounced influences than climate and soil chemical properties. PMID- 18430005 TI - Interactions among fungal community structure, litter decomposition and depth of water table in a cutover peatland. AB - Peatlands are important reservoirs of carbon (C) but our understanding of C cycling on cutover peatlands is limited. We investigated the decomposition over 18 months of five types of plant litter (Calluna vulgaris, Eriophorum angustifolium, Eriophorum vaginatum, Picea sitchensis and Sphagnum auriculatum) at a cutover peatland in Scotland, at three water tables. We measured changes in C, nitrogen (N) and phosphorus (P) in the litter and used denaturing gradient gel electrophoresis to investigate changes in fungal community composition. The C content of S. auriculatum litter did not change throughout the incubation period whereas vascular plant litters lost 30-40% of their initial C. There were no differences in C losses between low and medium water tables, but losses were always significantly less at the high water table. Most litters accumulated N and E. angustifolium accumulated significant quantities of P. C, N and P were significant explanatory variables in determining changes in fungal community composition but explained <25% of the variation. Litter type was always a stronger factor than water table in determining either fungal community composition or turnover of C, N and P in litter. The results have implications for the ways restoration programmes and global climate change may impact upon nutrient cycling in cutover peatlands. PMID- 18430006 TI - Cell-specific beta-N-acetylglucosaminidase activity in cultures and field populations of eukaryotic marine phytoplankton. AB - It is widely appreciated that eukaryotic marine phytoplankton can hydrolyze a variety of compounds within the dissolved organic matter (DOM) pool in marine environments. Herein, cultures and field populations of marine phytoplankton were assayed for beta-N-acetylglucosaminidase activity, a terminal enzyme of chitin degradation. A traditional bulk assay, which can assess hydrolytic rate, but is not cell-specific, was complemented with a cell-specific assay that images the activity associated with single cells using an enzyme labeled fluorescence (ELF) substrate. beta-N-acetylglucosaminidase activity was widespread across various taxa of marine phytoplankton, and activity was observed both under controlled culture conditions and in field populations. The number of cells with enzyme activity varied with the nutritional physiology of the test species in three of the 17 cultures tested. In these three cases the number of cells with activity in the low nutrient medium was higher than in nutrient replete medium. Taken together, these data suggest that a broad group of marine phytoplankton may be a relevant part of chitin-like DOM degradation and should be incorporated into conceptual models of chitin cycling in marine systems. PMID- 18430007 TI - In vitro fermentation of oat and barley derived beta-glucans by human faecal microbiota. AB - Fermentation of beta-glucan fractions from barley [average molecular mass (MM), of 243, 172, and 137 kDa] and oats (average MM of 230 and 150 kDa) by the human faecal microbiota was investigated. Fractions were supplemented to pH-controlled anaerobic batch culture fermenters inoculated with human faecal samples from three donors, in triplicate, for each substrate. Microbiota changes were monitored by fluorescent in situ hybridization; groups enumerated were: Bifidobacterium genus, Bacteroides and Prevotella group, Clostridium histolyticum subgroup, Ruminococcus-Eubacterium-Clostridium (REC) cluster, Lactobacillus Enterococcus group, Atopobium cluster, and clostridial cluster IX. Short-chain fatty acids and lactic acid were measured by HPLC. The C. histolyticum subgroup increased significantly in all vessels and clostridial cluster IX maintained high populations with all fractions. The Bacteroides-Prevotella group increased with all but the 243-kDa barley and 230-kDa oat substrates. In general beta-glucans displayed no apparent prebiotic potential. The SCFA profile (51 : 32 : 17; acetate : propionate : butyrate) was considered propionate-rich. In a further study a beta-glucan oligosaccharide fraction was produced with a degree of polymerization of 3-4. This fraction was supplemented to small-scale faecal batch cultures and gave significant increases in the Lactobacillus-Enterococcus group; however, the prebiotic potential of this fraction was marginal compared with that of inulin. PMID- 18430008 TI - Microbial primary production on an Arctic glacier is insignificant in comparison with allochthonous organic carbon input. AB - Cryoconite holes are unique freshwater environments on glacier surfaces, formed when solar-heated dark debris melts down into the ice. Active photoautotrophic microorganisms are abundant within the holes and fix inorganic carbon due to the availability of liquid water and solar radiation. Cryoconite holes are potentially important sources of organic carbon to the glacial ecosystem, but the relative magnitudes of autochthonous microbial primary production and wind-borne allochthonous organic matter brought are unknown. Here, we compare an estimate of annual microbial primary production in 2006 on Werenskioldbreen, a Svalbard glacier, with the organic carbon content of cryoconite debris. There is a great disparity between annual primary production (4.3 mug C g(-1) year(-1)) and the high content of organic carbon within the debris (1.7-4.5%, equivalent to 8500-22 000 mug C g(-1) debris). Long-term accumulation of autochthonous organic matter is considered unlikely due to ablation dynamics and the surface hydrology of the glacier. Rather, it is more likely that the majority of the organic matter on Werenskioldbreen is allochthonous. Hence, although glacier surfaces can be a significant source of organic carbon for glacial environments on Svalbard, they may be reservoirs rather than oases of high productivity. PMID- 18430009 TI - Phytophthora parasitica biofilm formation: installation and organization of microcolonies on the surface of a host plant. AB - Zoospores of the oomycete Phytophthora parasitica establish microbial spheroid microcolonies and biofilms on the surface of wounded leaves of their host, Nicotiana tabacum. The formation of microcolonies involves the movement of some zoospores towards attractants from wound sites, followed by their irreversible adsorption and the formation of a cluster of cells. These cells drive the migration of a second wave of zoospores (several hundreds cells) by setting up an external chemotactic gradient leading to massive zoospore encystment and cyst orientated germination. Zoospores that are still swimming at this stage circulate within the nascent biofilm by opening channels. Concomitantly, the cell population secretes various substances to elaborate an extracellular mucilage. Embedded within the extracellular matrix, biofilm cells are organized into a structured community as coacervates. The granular surface is composed of individual cysts, located on the outside of the microcolony. Hyphae from these cysts plunge downwards towards the dense core formed by the founder cells. This report is the first to show the installation and organization of a biofilm formed by eukaryotic cells on plant surfaces. The P. parasitica microcolonies constitute heterogeneous microenvironments for the embedded and circulating cells. They may affect plant-pathogen interactions by serving as reservoirs for pathogenic microorganisms, as protecting niche against host defences or as structures for infecting populations. PMID- 18430010 TI - Distribution of aerobic anoxygenic phototrophs in temperate freshwater systems. AB - The presence of aerobic anoxygenic phototrophs (AAPs) was recently reported from various marine environments; however, there is little information regarding their distribution in fresh waters. We surveyed a number of freshwater systems in central Europe, by infra-red fluorometry, infra-red epifluorescence microscopy, fluorescence emission spectroscopy and pigment analyses. AAPs were found to be abundant in several oligotrophic and mesotrophic lakes (50-400 ng of bacteriochlorophyll a l(-1), 10-80% of bacterial biomass), while in more eutrophized water bodies they represented a negligible part of the total microbial community (< 1%). The observed freshwater AAPs were morphologically diverse and different from previously observed marine species. Under temperate European climatic conditions, AAP populations undergo strong seasonal changes in terms of both abundance and species composition, with the maximum biomass in summer and the minimum in winter. In the mountain lakes Certovo and Plesne, AAPs contributed more than one half of total bacterial biomass during their summer maximum. These results show that photoheterotrophic bacteria represent an important part of the microbial community in many freshwater systems. PMID- 18430011 TI - Ammonia-oxidizing archaea: important players in paddy rhizosphere soil? AB - The diversity (richness and community composition) of ammonia-oxidizing archaea (AOA) and bacteria (AOB) in paddy soil with different nitrogen (N) fertilizer amendments for 5 weeks were investigated using quantitative real-time polymerase chain reaction, denaturing gradient gel electrophoresis (DGGE) jand clone library analysis based on the ammonia monooxygenase alpha-subunit (amoA) gene. Ammonia oxidizing archaea predominated among ammonia-oxidizing prokaryotes in the paddy soil, and the AOA:AOB DNA-targeted amoA gene ratios ranged from 1.2 to 69.3. Ammonia-oxidizing archaea were more abundant in the rhizosphere than in bulk soil. Rice cultivation led to greater abundance of AOA than AOB amoA gene copies and to differences in AOA and AOB community composition. These results show that AOA is dominant in the rhizosphere paddy soil in this study, and we assume that AOA were influenced more by exudation from rice root (e.g. oxygen, carbon dioxide) than AOB. PMID- 18430012 TI - Field-based and laboratory stable isotope probing surveys of the identities of both aerobic and anaerobic benzene-metabolizing microorganisms in freshwater sediment. AB - Laboratory incubations of coal-tar waste-contaminated sediment microbial communities under relatively controlled physiological conditions were used to interpret results of a field-based stable isotope probing (SIP) assay. Biodegradation activity of 13C-benzene was examined by GC/MS determination of net 13CO2 production and by GC headspace analysis of benzene loss. Key experimental variables were: the site of the assays (laboratory serum-bottle incubations and in situ field sediments), benzene concentration (10, 36 or 200 p.p.m. in laboratory assays), and physiological conditions (anaerobic with or without sulfate or nitrate additions versus aerobic headspace or the uncontrolled field). In anaerobic laboratory incubations of benzene at 10 p.p.m., greater than 60% of the substrate was eliminated within 15 days. During anaerobic incubations of 200 p.p.m. benzene (70 days), 0.9% benzene mineralization occurred. When benzene (36 p.p.m.) was added to sediment with air in the serum-bottle headspace, 14% of the initial 13C was mineralized to 13CO2 in 2.5 days. In the field experiment (178 microg 13C-benzene dosed to undisturbed sediments), net 13CO2 production reached 0.3% within 8.5 h. After isopycnic separation of 13C (heavy)-labelled DNA from the above biodegradation assays, sequencing of 13C-DNA clone libraries revealed a broad diversity of taxa involved in benzene metabolism and distinctive libraries for each biodegradation treatment. Perhaps most importantly, in the field SIP experiment the clone libraries produced were dominated by Pelomonas (betaproteobacteria) sequences similar to those found in the anaerobic 10 p.p.m. benzene laboratory experiment. These data indicate that the physiological conditions that prevail and govern in situ biodegradation of pollutants in the field may be interpreted by knowing the physiological preferences of potentially active populations. PMID- 18430013 TI - A pyrene-degrading consortium from deep-sea sediment of the West Pacific and its key member Cycloclasticus sp. P1. AB - A pyrene-degrading bacterial consortium was obtained from deep-sea sediments of the Pacific Ocean. The consortium degraded many kinds of polycyclic aromatic hydrocarbons (PAHs), including naphthalene, phenanthrene, pyrene, acenaphthene, fluorene, anthracene, fluoranthene, 2-methylnaphthalene and 2,6 dimethylnaphthalene, but it did not grow with chrysene and benzo[alpha]pyrene. With methods of plate cultivation and polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE), 72 bacteria belonging to 22 genera were detected from this consortium. Among the detected bacteria, the following genera frequently occurred: Flavobacterium, Cycloclasticus, Novosphingobium, Halomonas, Achromobacter, Roseovarius and Alcanivorax. The first two genera showed the strongest bands in denaturing gradient gel electrophoresis (DGGE) profiles and appeared in all PAH treatments. By now, only one isolate designated P1 was confirmed to be a pyrene degrader. It was identified to be Cycloclasticus spirillensus (100%). Although P1 can degrade pyrene independently, other bacteria, such as Novosphingobium sp. (Band 14), Halomonas sp. (Band 16) and an unidentified bacterium (Band 35), were involved in pyrene degradation in some way; they persist in the consortium in the test of dilution to extinction if only the consortium was motivated with pyrene. However, the secondary most important member Flavobacterium sp. evaded from the community at high dilutions. As a key member of the consortium, P1 distinguished itself by both cell morphology and carbon source range among the isolates of this genus. Based on intermediate analyses of pyrene degradation, P1 was supposed to take an upper pathway different from that previously reported. Together with the results of obtained genes from P1 homology with those responsible for naphthalene degradation, its degradation to pyrene is supposed to adopt another set of genes unique to presently detected. Summarily, an efficient pyrene-degrading consortium was obtained from the Pacific Ocean sediment, in which Cycloclasticus bacterium played a key role. This is the first report to exploit the diversity of pyrene degrading bacteria in oceanic environments. PMID- 18430014 TI - A novel, multi-layered methanotrophic microbial mat system growing on the sediment of the Black Sea. AB - A novel microbially diverse type of 1- to 5-cm-thick mat performing anaerobic oxidation of methane (AOM) and covering several square metres of the seafloor was discovered in the Black Sea at 180 m water depth. Contrary to other AOM-mat systems of the Black Sea these floating mats are not associated to free gas and are not stabilized by authigenic carbonates. However, supply of methane is ensured by the horizontal orientation of the mats acting as a cover of methane enriched fluids ascending from the underlying sediments. Thorough investigation of their community composition by molecular microbiology and lipid biomarkers, metabolic activities and elemental composition showed that the mats provide a clearly structured system with extracellular polymeric substances (EPS) building the framework of the mats. The top black zone, showing high rates of AOM (15 mumol g(dw) (-1) day(-1)), was dominated by ANME-2, while the following equally active pink layer was dominated by ANME-1 Archaea. The lowest AOM activity (2 mumol g(dw) (-1) day(-1)) and cell numbers were found in the greyish middle part delimited towards the sediment by a second pink, ANME-1-dominated and sometimes a black outer layer (ANME-2). Our work clearly shows that the different microbial populations are established along defined chemical gradients such as methane, sulfate or sulfide. PMID- 18430015 TI - Wide genetic diversity of picoplanktonic green algae (Chloroplastida) in the Mediterranean Sea uncovered by a phylum-biased PCR approach. AB - The genetic diversity of picoplanktonic (i.e. cells that can pass through a 3 mum pore-size filter) green algae was investigated in the Mediterranean Sea in late summer by a culture-independent approach. Genetic libraries of the 18S rRNA gene were constructed using two different primer sets. The first set is commonly used to amplify the majority of eukaryotic lineages, while the second was composed of a general eukaryotic forward primer and a reverse primer biased towards the phylum Chloroplastida. A total of 3980 partial environmental sequences were obtained: 1668 using the general eukaryotic primer set and 2312 using the Chloroplastida-biased primer set. Of these sequences, 65 (4%) and 594 (26%) belonged to the Chloroplastida respectively. A 99.5% sequence similarity cut-off value allowed classification of these 659 Chloroplastida sequences into 74 different operational taxonomic units. A majority of the Chloroplastida sequences (99%) belonged to the prasinophytes. In addition to the seven independent prasinophyte lineages previously described, we discovered two new clades (clades VIII and IX), as well as a significant genetic diversity at the species and subspecies levels, notably among the genera Crustomastix, Dolichomastix and Mamiella (Mamiellales), but also within Pyramimonas and Halosphaera (Pyramimonadales). Such diversity within prasinophytes has not previously been observed by cloning approaches, illustrating the power of using targeted primers for clone library construction. Prasinophyte assemblages differed especially in relation to nutrient levels. Micromonas and Ostreococcus were mainly recovered from mesotrophic areas, whereas Mamiella, Crustomastix and Dolichomastix were mostly detected in oligotrophic surface waters. Within genera such as Ostreococcus or Crustomastix for which several clades were observed, depth seemed to be the main factor controlling differential distribution of genotypes. PMID- 18430016 TI - Spatio-temporal niche separation of planktonic Betaproteobacteria in an oligo mesotrophic lake. AB - We investigated the diversity of planktonic Betaproteobacteria and the seasonal population changes of betaproteobacterial taxa in an oligo-mesotrophic lake (Piburger See, Austria). Focus was put on the vertical distribution of the investigated populations and on differences between their respective cell fractions with apparent amino acid incorporation. On average, 66% of betaproteobacterial cells and 73% of their diversity could be attributed to four clades within three lineages that were further analysed by fluorescence in situ hybridization. The numbers of bacteria from the R-BT subclade of the beta I lineage and from the PnecB subgroup of the beta II lineage were rather constant throughout the water column. In contrast, members of another subgroup of beta II (PnecC) and bacteria related to Methylophilus (beta IV) were particularly numerous in the oxygen-depleted zone. In general, only moderate seasonal changes in abundance were observed in the upper water layers, whereas there was a clear relationship between decreasing oxygen levels and the rise of bacteria from the PnecC and beta IV clades in deeper strata. On average, almost 80% of beta I bacteria, but < 15% of cells from the beta IV clade, showed amino acid incorporation. Our results suggest that the studied populations occupy distinct vertical and ecophysiological niches in Piburger See. PMID- 18430017 TI - Light affects motility and infectivity of Agrobacterium tumefaciens. AB - Response to changes in light conditions involves a variety of receptors that can modulate gene expression, enzyme activity and/or motility. For the study of light regulated effects of Agrobacterium tumefaciens, we used a global analysis approach - proteomics - and compared the protein patterns of dark- and light grown bacteria. These analyses revealed a significant reduction of FlaA and FlaB proteins of the flagellum - when the cells were grown in light. The light effect was confirmed by SDS-PAGE with isolated flagella. Quantitative PCR experiments showed a 10-fold increase of the transcription level of flaA, flaB and flaC within 20 min after the transfer from light to darkness. Electron microscopy revealed that these molecular events result in a light-induced reduction of the number of flagella per cell. These changes have major physiological consequences regarding motility, which is considerably reduced with exposure to light. The inhibitory effect of light on the motility is not unique to A. tumefaciens and was also seen in other species of the Rhizobiaceae. Previous studies suggested that the flagella function is significant for bacteria-plant interactions and bacterial virulence. In our studies, light reduced the attachment of A. tumefaciens to tomato roots and the virulence of the bacteria in a cucumber infection assay. PMID- 18430018 TI - Diversity of microbes associated with the marine sponge, Haliclona simulans, isolated from Irish waters and identification of polyketide synthase genes from the sponge metagenome. AB - Samples of the sponge Haliclona simulans were collected from Irish waters and subjected to a culture-independent analysis to determine the microbial, polyketide synthase (PKS) and non-ribosomal peptide synthase (NRPS) diversity. 16S rRNA gene libraries were prepared from total sponge, bacterial enriched sponge and seawater samples. Eight phyla from the Bacteria were detected in the sponge by phylogenetic analyses of the 16S rRNA gene libraries. The most abundant phylum in the total sponge library was the Proteobacteria (86%), with the majority of these clones being from the gamma-Proteobacteria (77%); two groups of clones were dominant and together made up 69% of the total. Both of these groups were related to other sponge-derived microbes and comprised novel genera. Within the other bacterial phyla groups of clones representing novel candidate genera within the phyla Verrucomicrobia and Lentisphaerae were also found. Selective enrichment of the bacterial component of the sponge prior to 16S rRNA gene analysis resulted in a 16S rRNA gene library dominated by a novel genus of delta Proteobacteria, most closely related to the Bdellovibrio. The potential for the sponge microbiota to produce secondary metabolites was also analysed by polymerase chain reaction amplification of PKS and NRPS genes. While no NRPS sequences were isolated seven ketosynthase (KS) sequences were obtained from the sponge metagenome. Analyses of these clones revealed a diverse collection of PKS sequences which were most closely affiliated with PKS from members of the Cyanobacteria, Myxobacteria and Dinoflagellata. PMID- 18430019 TI - Diel rhythmicity in amino acid uptake by Prochlorococcus. AB - The marine cyanobacterium Prochlorococcus, the most abundant phototrophic organism on Earth, numerically dominates the phytoplankton in nitrogen (N) depleted oceanic gyres. Alongside inorganic N sources such as nitrite and ammonium, natural populations of this genus also acquire organic N, specifically amino acids. Here, we investigated using isotopic tracer and flow cytometric cell sorting techniques whether amino acid uptake by Prochlorococcus is subject to a diel rhythmicity, and if so, whether this was linked to a specific cell cycle stage. We observed, in contrast to diurnally similar methionine uptake rates by Synechococcus cells, obvious diurnal rhythms in methionine uptake by Prochlorococcus cells in the tropical Atlantic. These rhythms were confirmed using reproducible cyclostat experiments with a light-synchronized axenic Prochlorococcus (PCC9511 strain) culture and (35)S-methionine and (3)H-leucine tracers. Cells acquired the tracers at lower rates around dawn and higher rates around dusk despite >10(4) times higher concentration of ammonium in the medium, presumably because amino acids can be directly incorporated into protein. Leucine uptake rates by cells in the S+G(2) cell cycle stage were consistently 2.2 times higher than those of cells at the G(1) stage. Furthermore, S+G(2) cells upregulated amino acid uptake 3.5 times from dawn to dusk to boost protein synthesis prior to cell division. Because Prochlorococcus populations can account from 13% at midday to 42% at dusk of total microbial uptake of methionine and probably of other amino acids in N-depleted oceanic waters, this genus exerts diurnally variable, strong competitive pressure on other bacterioplankton populations. PMID- 18430020 TI - Use of the rotating wall vessel technology to study the effect of shear stress on growth behaviour of Pseudomonas aeruginosa PA01. AB - The biofilm phenotype of Pseudomonas aeruginosa enables this opportunistic pathogen to develop resistance to the immune system and antimicrobial agents. Pseudomonas aeruginosa biofilms are generated under varying levels of shear stress, depending on the infection site. In the lung mucus of cystic fibrosis (CF) patients, P. aeruginosa forms matrix-enclosed microcolonies which cause chronic infections representing the major cause of mortality in CF patients. The lung mucus of CF patients is probably characterized by low fluid shear as the main shear-causing factor, i.e. mucociliary clearance, is absent. In this study, the influence of fluid shear on the growth behaviour of P. aeruginosa PA01 was investigated using a low-shear suspension culture device, the rotating wall vessel (RWV). Cultivation in low shear induced a self-aggregating phenotype of P. aeruginosa PA01, resulting in the formation of biofilms in suspension similar to what has been described in CF mucus. The addition of a ceramic bead to the culture medium in the RWV created a higher-shear condition which led to the formation of surface-attached rather than suspension biofilms. In low-shear culture conditions, a significant increase of the rhl N-butanoyl-l-homoserine lactone (C(4)-HSL) directed quorum sensing (QS) system, and the psl polysaccharide synthetic locus was demonstrated using gene expression analysis. Accordingly, the low-shear condition induced a higher production of rhamnolipids, which is controlled by the C(4)-HSL QS-system and is known to play a role in CF lung pathology. These results indicate that fluid shear has an impact on the growth phenotype of P. aeruginosa which might play a role in CF lung infections caused by this bacterium. PMID- 18430022 TI - Genome-wide identification, classification, evolutionary expansion and expression analyses of homeobox genes in rice. AB - Homeobox genes play a critical role in regulating various aspects of plant growth and development. In the present study, we identified a total of 107 homeobox genes in the rice genome and grouped them into ten distinct subfamilies based upon their domain composition and phylogenetic analysis. A significantly large number of homeobox genes are located in the duplicated segments of the rice genome, which suggests that the expansion of homeobox gene family, in large part, might have occurred due to segmental duplications in rice. Furthermore, microarray analysis was performed to elucidate the expression profiles of these genes in different tissues and during various stages of vegetative and reproductive development. Several genes with predominant expression during various stages of panicle and seed development were identified. At least 37 homeobox genes were found to be differentially expressed significantly (more than two-fold; P < 0.05) under various abiotic stress conditions. The results of the study suggest a critical role of homeobox genes in reproductive development and abiotic stress signaling in rice, and will facilitate the selection of candidate genes of agronomic importance for functional validation. PMID- 18430021 TI - Microbial community analysis of two field-scale sulfate-reducing bioreactors treating mine drainage. AB - The microbial communities of two field-scale pilot sulfate-reducing bioreactors treating acid mine drainage (AMD), Luttrell and Peerless Jenny King (PJK), were compared using biomolecular tools and multivariate statistical analyses. The two bioreactors were well suited for this study because their geographic locations and substrate compositions were similar while the characteristics of influent AMD, configuration and degree of exposure to oxygen were distinct. The two bioreactor communities were found to be functionally similar, including cellulose degraders, fermenters and sulfate-reducing bacteria (SRB). Significant differences were found between the two bioreactors in phylogenetic comparisons of cloned 16S rRNA genes and adenosine 5'-phosphosulfate reductase (apsA) genes. The apsA gene clones from the Luttrell bioreactor were dominated by uncultured SRB most closely related to Desulfovibrio spp., while those of the PJK bioreactor were dominated by Thiobacillus spp. The fraction of the SRB genus Desulfovibrio was also higher at Luttrell than at PJK as determined by quantitative real-time polymerase chain reaction analysis. Oxygen exposure at PJK is hypothesized to be the primary cause of these differences. This study is the first rigorous phylogenetic investigation of field-scale bioreactors treating AMD and the first reported application of multivariate statistical analysis of remediation system microbial communities applying UniFrac software. PMID- 18430023 TI - Hatching enzyme of the ovoviviparous black rockfish Sebastes schlegelii- environmental adaptation of the hatching enzyme and evolutionary aspects of formation of the pseudogene. AB - The hatching enzyme of oviparous euteleostean fishes consists of two metalloproteases: high choriolytic enzyme (HCE) and low choriolytic enzyme (LCE). They cooperatively digest the egg envelope (chorion) at the time of embryo hatching. In the present study, we investigated the hatching of embryos of the ovoviviparous black rockfish Sebastes schlegelii. The chorion-swelling activity, HCE-like activity, was found in the ovarian fluid carrying the embryos immediately before the hatching stage. Two kinds of HCE were partially purified from the fluid, and the relative molecular masses of them matched well with those deduced from two HCE cDNAs, respectively, by MALDI-TOF MS analysis. On the other hand, LCE cDNAs were cloned; however, the ORF was not complete. These results suggest that the hatching enzyme is also present in ovoviviparous fish, but is composed of only HCE, which is different from the situation in other oviparous euteleostean fishes. The expression of the HCE gene was quite weak when compared with that of the other teleostean fishes. Considering that the black rockfish chorion is thin and fragile, such a small amount of enzyme would be enough to digest the chorion. The black rockfish hatching enzyme is considered to be well adapted to the natural hatching environment of black rockfish embryos. In addition, five aberrant spliced LCE cDNAs were cloned. Several nucleotide substitutions were found in the splice site consensus sequences of the LCE gene, suggesting that the products alternatively spliced from the LCE gene are generated by the mutations in intronic regions responsible for splicing. PMID- 18430024 TI - Purification and characterization of organellar DNA polymerases in the red alga Cyanidioschyzon merolae. AB - DNA polymerase gamma, a mitochondrial replication enzyme of yeasts and animals, is not present in photosynthetic eukaryotes. Recently, DNA polymerases with distant homology to bacterial DNA polymerase I were reported in rice, Arabidopsis, and tobacco, and they were localized to both plastids and mitochondria. We call them plant organellar DNA polymerases (POPs). However, POPs have never been purified in the native form from plant tissues. The unicellular thermotrophic red alga Cyanidioschyzon merolae contains two genes encoding proteins related to Escherichia coli DNA polymerase I (PolA and PolB). Phylogenetic analysis revealed that PolB is an ortholog of POPs. Nonphotosynthetic eukaryotes also have POPs, which suggested that POPs have an ancient origin before eukaryotic photosynthesis. PolA is a homolog of bacterial DNA polymerase I and is distinct from POPs. PolB was purified from the C. merolae cells by a series of column chromatography steps. Recombinant protein of PolA was also purified. Sensitivity to inhibitors of DNA synthesis was different in PolA, PolB, and E. coli DNA polymerase I. Immunoblot analysis and targeting studies with green fluorescent protein fusion proteins demonstrated that PolA was localized in the plastids, whereas PolB was present in both plastids and mitochondria. The expression of PolB was regulated by the cell cycle. The available results suggest that PolB is involved in the replication of plastids and mitochondria. PMID- 18430025 TI - Role of disulfide bonds in goose-type lysozyme. AB - The role of the two disulfide bonds (Cys4-Cys60 and Cys18-Cys29) in the activity and stability of goose-type (G-type) lysozyme was investigated using ostrich egg white lysozyme as a model. Each of the two disulfide bonds was deleted separately or simultaneously by substituting both Cys residues with either Ser or Ala. No remarkable differences in secondary structure or catalytic activity were observed between the wild-type and mutant proteins. However, thermal and guanidine hydrochloride unfolding experiments revealed that the stabilities of mutants lacking one or both of the disulfide bonds were significantly decreased relative to those of the wild-type. The destabilization energies of mutant proteins agreed well with those predicted from entropic effects in the denatured state. The effects of deleting each disulfide bond on protein stability were found to be approximately additive, indicating that the individual disulfide bonds contribute to the stability of G-type lysozyme in an independent manner. Under reducing conditions, the thermal stability of the wild-type was decreased to a level nearly equivalent to that of a Cys-free mutant (C4S/C18S/C29S/C60S) in which all Cys residues were replaced by Ser. Moreover, the optimum temperature of the catalytic activity for the Cys-free mutant was downshifted by about 20 degrees C as compared with that of the wild-type. These results indicate that the formation of the two disulfide bonds is not essential for the correct folding into the catalytically active conformation, but is crucial for the structural stability of G-type lysozyme. PMID- 18430027 TI - Application of heat shock promoter in transgenic zebrafish. AB - The heat shock promoter is useful for regulating transgene expression in small water-living organisms. In zebrafish embryos, downstream gene expression can be greatly induced throughout the body by raising the temperature from 28.5 degrees C to 38.0 degrees C. By manipulating the local temperature within an embryo, spatial control of transgene expression is also possible. One such way for inducing heat shock response in targeted cells is by using a laser microbeam under the microscope. In addition, random mosaic expression by transient gene expression and transplantation of the transgenic embryo into a wild type host can be considered a powerful tool for studying gene functions using this promoter. In this paper, we review the applications of the zebrafish heat shock protein promoter as a gene expression tool and for lineage labeling and transcription enhancer screening. PMID- 18430028 TI - Bone development in the jaw of Nile tilapia Oreochromis niloticus (Pisces: Cichlidae). AB - East African cichlids have evolved feeding apparatus morphologies adapted to their diverse feeding behaviors. The evolution of the oral jaw morphologies is accomplished by the diversity of bone formation during development. To further understand this evolutionary process, we examined the skeletal elements of the jaw and their temporal and sequential emergence, categorized by developmental stages, using the Nile tilapia Oreochromis niloticus as a model cichlid. We found that chondrogenesis started in Stage 17. The deposition of osteoid for the dermal bones commenced in Stage 18. The uptake of calcium dramatically shifted from the surface of larvae to the gills in Stage 20. The bone mineralization of the skeleton began in Stage 25. These data provide important information regarding the sequential events of craniofacial development in East African cichlids and lay the groundwork for studying the molecular mechanisms underlying adaptation of jaw structure to feeding behavior. PMID- 18430026 TI - Proper Gcn5 histone acetyltransferase expression is required for normal anteroposterior patterning of the mouse skeleton. AB - Histone acetylation plays important roles in gene regulation. However, the functions of individual histone acetyltransferases (HATs) in specific developmental transcription programs are not well defined. To define the functions of Gcn5, a prototypical HAT, during mouse development, we have created a series of mutant Gcn5 alleles. Our previous work revealed that deletion of Gcn5 leads to embryonic death soon after gastrulation. Embryos homozygous for point mutations in the catalytic center of Gcn5 survive longer, but die soon after E16.0 and exhibit defects in cranial neural tube closure. Embryos bearing a hypomorphic Gcn5(flox(neo)) allele also exhibit neural closure defects and die at or soon after birth. We report here that Gcn5(flox(neo)/flox(neo)) and Gcn5(flox(neo)/Delta) embryos exhibit anterior homeotic transformations in lower thoracic and lumbar vertebrae. These defects are accompanied by a shift in the anterior expression boundary of Hoxc8 and Hoxc9. These data provide the first evidence that Gcn5 contributes to Hox gene regulation and is required for normal anteroposterior patterning of the mouse skeleton. PMID- 18430029 TI - Gene transduction by sonoporation. AB - Gene transduction technologies are essential tools for understanding of gene functions or gene cascades underlying embryogenesis. In this review, we introduce a gene transduction method using microbubble and ultrasound (hereafter referred to as sonoporation). Sonoporation is carried out with relatively simple procedures and easily transduces genes into mesenchymal cells without significant damage to target tissues. Therefore, sonoporation is effective for gene transduction to study the molecular mechanisms of morphogenesis. PMID- 18430030 TI - Differential effects of pro-BDNF on sensory neurons after sciatic nerve transection in neonatal rats. AB - Brain-derived neurotrophic factor (BDNF) plays a critical role in the development of the central and peripheral nervous systems, and also in neuronal survival after injury. The actions of BDNF are mediated by its high-affinity receptors TrkB and p75NTR. Recent studies have shown that proneurotrophins bind p75NTR and sortilin with high affinity, and trigger apoptosis of neurons in vitro. As proneurotrophins are a dominant form of gene products in developing and adult animals, it is imperative to understand their physiological functions in animals. Here, we showed differential roles of proBDNF in injured and uninjured sensory neurons. proBDNF, p75NTR and sortilin are highly expressed in dorsal root ganglia (DRG) neurons. Recombinant proBDNF induced a dose-dependent death of PC12 cells and the death activity was completely abolished in the presence of antibodies against the prodomain of BDNF. The exogenous proBDNF enhanced the death of axotomized sensory neurons and the neutralizing antibodies to the prodomain or exogenous sortilin-extracellular domain-Fc fusion molecule reduced the death of axotomized sensory neurons. Interestingly, the treatment of neutralizing antibody in vivo increased the number of sensory neurons in the contralateral DRG. We conclude that proBDNF may induce the death of axotomized sensory neurons and suppress neuronal addition in the intact DRG in neonatal rats, and the suppression of endogenous proBDNF may protect neurons after neurotrauma. PMID- 18430031 TI - Movement-related synchronization of gamma activity is lateralized in patients with dystonia. AB - There is evidence for synchronization at frequencies both under 30 Hz and over 60 80 Hz in the so-called gamma frequency band in patients with Parkinson's disease (PD). Gamma activity increases after dopaminergic therapy and during voluntary movement, suggesting that it might be physiological and relate to motor processing in the basal ganglia (BG). We recorded local field potential (LFP) activity during a choice reaction time task in 11 patients with dystonia undergoing implantation of the internal globus pallidus for therapeutic stimulation. The spectral content of the LFP was averaged with respect to movement onset over 6-11 Hz, 18-25 Hz and 60-80 Hz, separately for responses ipsilateral and contralateral to movement. There was a perimovement increase in 60-80 Hz activity in the LFP, but only contralateral to movement. In contrast, low-frequency LFP activity decreased symmetrically during movement. This occurred earlier in the 18-25 Hz band than in the 6-11 Hz band, and was followed by a postmovement increase in oscillatory activity in the 18-25 Hz band that was contralateral to movement. The presence of a lateralized movement-related increase in gamma activity in the BG of patients with dystonia, similar to that recorded in patients with treated PD, suggests that this may be a residual feature of normal BG function. Moreover, the results provide further support for functional distinctions between BG oscillatory activities of different frequency. PMID- 18430032 TI - Role of GluR1 expression in nucleus accumbens neurons in cocaine sensitization and cocaine-seeking behavior. AB - Chronic cocaine use reduces glutamate levels in the nucleus accumbens (NAc), and is associated with experience-dependent changes in (+/-)-alpha-amino-3-hydroxy-5 methylisoxazole-4-propionic acid (AMPA) glutamate receptor membrane expression in NAc neurons. These changes accompany behavioral sensitization to cocaine and increased susceptibility to cocaine relapse. The functional relationship between neuroplasticity in AMPA receptors and the behavioral manifestation of cocaine addiction remains unclear. Thus, we examined the behavioral effects of up- and downregulating basal AMPA receptor function in the NAc core and shell using viral mediated gene transfer of wild-type glutamate receptor 1 (wt-GluR1) or a dominant negative pore-dead GluR1 (pd-GluR1), respectively. Transient increases in wt GluR1 during or after cocaine treatments diminished the development of cocaine sensitization, while pd-GluR1 expression exacerbated cocaine sensitization. Parallel changes were found in D2, but not D1, receptor-mediated behavioral responses. As a correlate of the sensitization experiments, we overexpressed wt- or pd-GluR1 in the NAc core during cocaine self-administration, and tested the effects on subsequent drug-seeking behavior 3 weeks after overexpression declined. wt-GluR1 overexpression during self-administration had no effect on cocaine intake, but subsequently reduced cocaine seeking in extinction and cocaine-induced reinstatement, whereas pd-GluR1 facilitated cocaine-induced reinstatement. When overexpressed during reinstatement tests, wt-GluR1 directly attenuated cocaine- and D2 agonist-induced reinstatement, while pd-GluR1 enhanced reinstatement. In both experimental procedures, neither wt- nor pd-GluR1 expression affected cue-induced reinstatement. Together, these results suggest that degrading basal AMPA receptor function in NAc neurons is sufficient to facilitate relapse via sensitization in D2 receptor responses, whereas elevating basal AMPA receptor function attenuates these behaviors. PMID- 18430033 TI - Pain influences hedonic assessment of visual inputs. AB - It is acknowledged that the emotional state created by visual inputs can modulate the way we feel pain; however, little is known about how acute pain influences the emotional assessment of what we see. In this study healthy subjects scored affective images while receiving painful or innocuous electrical shocks. Painful stimuli did not make unpleasant images more unpleasant, but rendered pleasant pictures significantly less pleasant. Brain responses to visual inputs (64 channels electroencephalogram) mirrored behavioural results, showing pain-induced effects in the orbitofrontal cortex, the subgenual portion of the cingulate gyrus, the anterior prefrontal and the temporal cortices, exclusively during presentation of pleasant images. In addition to this specific effect on pleasant pictures, pain also produced non-specific effects upon all categories of images, engaging cerebral areas associated with attention, alertness and motor preparation (middle-cingulate, supplemental motor, prefrontal cortex). Thus, pain appears to have a dual influence on visual processing: a non-specific effect related to orienting phenomena; and a more specific action exerted on supra-modal limbic areas involved in the production of affective states. The latter correlated with changes in the subjective appraisal of visual stimuli, and may underlie not only the change in their subjective assessment but also reactive processes aimed at coping with unpleasant contexts. PMID- 18430034 TI - Coincident pre- and postsynaptic activity downregulates NKCC1 to hyperpolarize E(Cl) during development. AB - In the mature CNS, coincident pre- and postsynaptic activity decreases the strength of gamma-aminobutyric acid (GABA)(A)-mediated inhibition through a Ca2+ dependent decrease in the activity of the neuron-specific K+-Cl- cotransporter KCC2. In the present study we examined whether coincident pre- and postsynaptic activity can also modulate immature GABAergic synapses, where the Na+-K+-2Cl- (NKCC1) cotransporter maintains a relatively high level of intracellular chloride ([Cl-](i)). Dual perforated patch-clamp recordings were made from cultured hippocampal neurons prepared from embryonic Sprague-Dawley rats. These recordings were used to identify GABAergic synapses where the reversal potential for Cl- (ECl) was hyperpolarized with respect to the action potential threshold but depolarized with respect to the resting membrane potential. At these synapses, repetitive postsynaptic spiking within +/- 5 ms of GABAergic synaptic transmission resulted in a hyperpolarizing shift of ECl by 10.03 +/- 1.64 mV, increasing the strength of synaptic inhibition. Blocking the inward transport of Cl- by NKCC1 with bumetanide (10 microm) hyperpolarized ECl by 16.14 +/- 4.8 mV, and prevented this coincident activity-induced shift of ECl. The bumetanide induced hyperpolarization of ECl occluded furosemide, a K+-Cl- cotransporter antagonist, from producing further shifts in ECl. Together, this indicates that brief coincident pre- and postsynaptic activity strengthens inhibition through a regulation of NKCC1. This study further demonstrates ionic plasticity as a mechanism underlying inhibitory synaptic plasticity. PMID- 18430035 TI - Fishing Down Under: case report and review of management of a fishhook injury of the eyelid. AB - A 21-year-old man presented to the emergency department of the Royal Victorian Eye and Ear Hospital with a fishhook embedded in his unprotected left upper eyelid. The fishhook was removed after exploration of the left eye by vertical eyelid incision. Subsequent eyelid reconstruction by lid margin adaptation was performed. Management of these injuries depends on type of hook, the involved ocular structure and location of the hook. This is an update on management options, where triage and surgical approaches are discussed. This case illustrates the risk to the eyes while fishing. Persons with an interest in fishing should be advised to wear eye protection. PMID- 18430036 TI - Dyslexia: a review of two theories. AB - Optometrists will frequently see patients, who may have a diagnosis or a suspected diagnosis of dyslexia (specific reading disorder) and will need to manage and counsel such patients. There are many propounded theories on the cause(s) of dyslexia. Although most professionals in this area consider that dyslexia is chiefly a linguistic disorder, the possibility of a visual component is contentious. This article is a selective review of two commonly discussed theories that suggest a visual component in dyslexia; the magnocellular deficit theory and Meares-Irlen syndrome. PMID- 18430037 TI - Aspects of wet cell measurement of back vertex power of contact lenses. AB - BACKGROUND: The back vertex power of soft contact lenses can be measured when they are immersed in saline within a wet cell. Six equations have been published for the purpose of converting the power measured in saline to its corresponding power in air. Results provided by these equations have not been compared and the consequences of inaccuracies in the variables used in them have not been quantified. METHODS: Sample data were used to solve these equations and inaccuracies were simulated in each of the variables required for their solution to determine the resultant magnitude of error in the calculated power in air. RESULTS: Three equations gave identical results. Inaccuracies in the lens specification, distance of the back vertex of the lens from the cell wall, refractive index of the cell and its wall thickness were of very little consequence in the calculation of power in air. In the case of a high water content lens, immersed power must be measured to 0.02 D and its refractive index and that of saline must be known to +/-0.001 to prevent errors in the calculated power in air exceeding tolerance limits. CONCLUSION: Measured power in saline can be converted to power in air using the original equation published by Wichterle in 1965. While the nominal specification of the lens can be used in this equation, it is essential to know its actual refractive index and that of the saline. Reproducibility of wet cell measurement of power using a suitable focimeter is unknown and should be established to allow this method to be used by practitioners and research laboratories. PMID- 18430038 TI - Bilateral macular sub-retinal fluid and retinal pigment epithelial detachment associated with type 2 membrano-proliferative glomerulonephritis. AB - Choroidal neovascularisation (CNV) and idiopathic central serous chorioretinopathy (ICSC) are recognised ocular complications related to type 2 membrano-proliferative glomerulonephritis. We report a 38-year-old white male who presented with a 10-day history of blurring of vision, micropsia and metamorphopsia. He had been diagnosed recently to have type 2 membrano proliferative glomerulonephritis. On examination, there was bilateral retinal pigment epithelial (RPE) detachment with overlying sub-retinal fluid without any drusen. Fundus fluorescein angiography (FFA) and optical coherence tomography (OCT) confirmed the diagnosis of atypical ICSC. Three months later, sub-retinal fluid and RPE detachment resolved and VA had recovered to 6/6. The case highlights the importance of ophthalmological assessment in these patients to recognise sight-threatening complications. PMID- 18430039 TI - Automated determination of distance visual acuity: towards teleophthalmology services. AB - PURPOSE: The aim of this study was to evaluate the visual acuity scores of a computer-based visual function testing (CVFT) system. The automated chart uses logMAR notation (logarithm of the minimum angle of resolution) and randomised tilting E as optotype. The test was conducted using a laptop computer. METHODS: This prospective, clinic-based, test validation study was based on a model of repeatability in two observations. The right eyes of a total of 104 participants were randomly tested with both CVFT and the manual, gold standard Early Treatment of Diabetic Retinopathy Study (ETDRS) chart. RESULTS: The mean difference between CVFT and ETDRS was -0.01+/-0.11 (SD) logMAR units. The Spearman Rank correlation coefficient (r) between the absolute difference and the average was 0.35. The average time for testing one eye was 54.7+/-11.89 seconds. The mean difference between two consecutive, repeated CVFT tests in the same eye was 0.02+/-0.07 logMAR units (paired t-test p=0.046). CONCLUSIONS: A novel, automated visual acuity testing application is reported. This automated system should be a useful tool for mass screening of visual impairment in rural and remote regions with its ease of use and shorter testing time. PMID- 18430040 TI - Task division between dentists and dental hygienists in Norway. AB - OBJECTIVE: The aim of the study was to investigate the attitudes among dentists and dental hygienists to the policy objective in Norway of delegating more dental work from dentists to dental hygienists. METHOD: A questionnaire was mailed to a random sample of 1111 dentists and 268 dental hygienists in 2005. The response rates were 45% (504) among the dentists and 42% (112) among the dental hygienists. The survey sought to explore any discrepancies between current and preferred mix of different work tasks, as well as attitudes to the idea of substituting dentists with dental hygienists for certain work tasks. Logistic regression was used to analyse how answers differed by respondent characteristics. RESULTS: Dentists spent only half of their total working hours on complex dental services, i.e. tasks that only dentists are skilled to undertake. Nearly 40% of their time was spent on tasks that dental hygienists are qualified to perform; examinations, screening and basic treatments. Still, the mix of work tasks that dentists preferred would involve slight changes: on average only 2% points more complex treatment and 3-4% points less of those tasks that dental hygienists are permitted to provide. Seemingly contrary, as many as 60% of dentists answered that it was 'desirable to delegate' more tasks to dental hygienist. However, only 21% of the dentists agreed that dental hygienists should be the entry point for dental services. Dental hygienists would prefer to do relatively more basic treatments and fewer examinations and screening, and the vast majority among them supported the idea that they could be the entry point for dental services. CONCLUSION: The results suggest that there will not be major changes in the division of labour between dentists and dental hygienists in Norway, if dentists are to be held responsible for taking such initiatives. Although dentists agree that more of their current work could- in principle - be delegated to dental hygienists, they do not prefer to reduce much of their own current activity of those work tasks that dental hygienists are qualified to perform. PMID- 18430041 TI - Nursing caries and buying time: an emerging theory of prolonged bottle feeding. AB - BACKGROUND: Nursing caries is considered to be problematic by dental health professionals. In their zealousness to solve the nursing caries problem dental health professionals forgot to ask the question: 'Why do mothers persist in prolonged bottle feeding?' AIM: To use grounded theory procedures and techniques to analyse the qualitative data obtained from mothers and to generate an emerging theory of prolonged bottle feeding. METHOD: A series of individual in-depth interviews were conducted with 34 mothers of children with nursing caries. The children were aged between 3 and 4 years. Data were analysed using the grounded theory procedures and techniques of open and selective coding. RESULTS: The core category or the mothers' main concern that emerged from the data was conceptualized as 'buying time'. The feeding bottle bought time away from crying children by silencing them but also bought extra time with the child. Therefore, two different behavioural styles or categories of buying time emerged. These were: (i) to buy time away from the demands of their crying toddlers (instant solutions) and (ii) to buy extra time with their 'toddler-babies' (double binding). The feeding bottle acted as an 'instant solution' as on seeing the bottle the child would instantly 'stop crying' and peace would reign. Double binding described how the feeding bottle was used to buy extra time for 'babyhood closeness' between mother and child while exposing the mothers' harsh rejecting behaviours. DISCUSSION: An understanding of the time concerns that the mothers experienced when caring for their young children and how they resolve them provides an important insight into the reasons for prolonged bottle feeding. PMID- 18430042 TI - Evaluation of a generic quality of life instrument for early childhood caries related pain. AB - OBJECTIVE: This prospective study evaluated the reliability and validity of the Infant and Toddler Child Quality of Life Questionnaire (ITQOL) by comparing quality of life (QOL) of two groups of children afflicted with early childhood dental caries with a caries-free group. METHODS: A convenience sample of 150 children divided into three groups (mean age [months] 51 +/- 15) was studied. Group I (OR) had extensive dental caries and were waiting for several months for treatment under general anesthesia. Group II were children who sought emergency services for acute odontogenic pain/abscess, and group III were children who were caries-free new patients. Parents/caretakers were administered the ITQOL, which assesses a child's physical, emotional and social well being from the parent/guardian's perspective. RESULTS: The ITQOL had high success in item validity with good reliability. Significant differences in QOL were noted for scales of pain and discomfort, behavior and moods, global behavior, and impact on parental time (P 0.05). After exercise, the decrease in diastolic BP was greater with hexamethonium (-26 +/- 1 mmHg; P < 0.05) compared with the decrease caused by exercise alone. 5. The results suggest that acute resistance exercise has an important hypotensive effect on SHR and that NO plays a crucial role in this response. PMID- 18430049 TI - Post-transfusional variation in urinary oxygen tension in surgical patients. AB - Prior studies have suggested urinary oxygen tension (P(u)O2) as a putative index of renal blood flow (RBF) and tissue oxygenation. In the present study, we collected intraoperative data from eight anaemic, bladder-catheterized patients who received erythrocyte transfusions during various surgical procedures under general anaesthesia. Urinary and arterial blood gas analysis and co-oximetry were performed before and after transfusion, during an interval in which RBF was assumed to be constant. Pre- and post-transfusion haematocrit levels were 23 +/- 4.8 and 27 +/- 6.4%, respectively, and arterial oxygen content (C(a)O2) was 10.5 +/- 2.5 and 12.4 +/- 3.1 mL/dL, respectively (P = 0.012). Pre- and post transfusion P(u)O2 was 90 +/- 14 and 108 +/- 20 mmHg, respectively (P = 0.036). 4. These results, although limited, suggest that under conditions of stable haemodynamics, systemic oxygenation and renal function, intraoperative blood transfusion may increase P(u)O2 in anaemic anaesthetized patients. If confirmed by subsequent clinical and laboratory studies, P(u)O2 measurement could become a simple, non-invasive way to monitor renal medullary oxygenation and tissue oxygen availability and help determine whether red blood cells should be transfused. PMID- 18430050 TI - The CGP7930 analogue 2,6-di-tert-butyl-4-(3-hydroxy-2-spiropentylpropyl)-phenol (BSPP) potentiates baclofen action at GABA(B) autoreceptors. AB - The pharmacological actions of 2,6-di-tert-butyl-4-(3-hydroxy-2 spiropentylpropyl)-phenol (BSPP), a putative presynaptic GABA(B) receptor modulator, were examined in electrically stimulated rat neocortical brain slices preloaded with [3H]-GABA or [3H]-glutamic acid. At 10 mmol/L, BSPP inhibited the release of [3H]-GABA in the presence of baclofen, but not that of [3H]-glutamic acid. This effect was sensitive to the GABA(B) receptor antagonist (+)-(S)-5,5 dimethylmorpholinyl-2-acetic acid (Sch 50911). Alone, BSPP had no effect on the release of [3H]-GABA or [3H]-glutamic acid. It is concluded that BSPP selectively potentiates the action of baclofen at GABA(B) autoreceptors, but not heteroreceptors and may be a useful ligand to discriminate between presynaptic GABA(B) receptor subtypes. PMID- 18430051 TI - N-Acetylcysteine prevents but does not reverse dexamethasone-induced hypertension. AB - 1. We have shown previously that N-acetylcysteine (NAC) prevents the increase in blood pressure induced by adrenocorticotropin treatment. The present study investigated the effect of NAC on dexamethasone (Dex)-induced hypertension. 2. Male Sprague-Dawley rats were randomly divided into six groups (n = 10 in each). In a prevention study, NAC (10 g/L in the drinking water) was given for 4 days prior to and 11 days during concurrent treatment with saline (0.1 mL/rat per day) or with Dex (10 mg/rat per day). In a reversal study, daily injections of Dex or saline began 8 days before NAC and cotreatment continued for 5 days. Systolic blood pressure (SBP) was measured on alternate days using a tail-cuff system. 3. Dexamethasone significantly increased SBP from 113 +/- 4 to 139 +/- 6 mmHg (n = 10; P < 0.01). N-Acetylcysteine alone had no effect on SBP. In NAC + Dex-treated rats, SBP was significantly lower than that of Dex-treated rats (P cent < 0.01). In fully established Dex-hypertension NAC was ineffective and SBP remained high. 4. Both Dex and NAC treatments decreased bodyweight gain. N-Acetylcysteine reduced food and water consumption. Dexamethasone reduced thymus weight (P cent < 0.01) but NAC treatment did not alter this marker of glucocorticoid activity. 5. Dexamethasone tended to decrease plasma NO(x), whereas NAC restored plasma NO(x) concentrations to control levels. N-Acetylcysteine had no effect on Dex-induced increased plasma F(2)-isoprostane concentrations. 6. In conclusion, NAC partially prevented, but did not reverse, Dex-induced hypertension. PMID- 18430052 TI - A comparison of the pharmacological properties of recombinant human and rat alpha(1)beta(2)gamma(2L) GABA(A) receptors in Xenopus oocytes. AB - In the present study, we compared the pharmacology, particularly neurosteroid modulation of the GABA(A) receptor, between human and rat alpha(1)beta(2)gamma(2)(L) GABA(A) receptors and between human receptors containing the long (L) and short (S) forms of the gamma(2)-subunit. We observed that maximum responses to GABA were significantly higher with the human alpha(1)beta(2)gamma(2)(L) receptor compared with the rat receptor. In terms of neurosteroid modulation, increases in the EC(15) response to GABA induced by 3alpha-OH-5beta-pregnan-20-one (3alpha5betaP), 5alpha-androstane-3alpha,17beta diol (3alpha5alphaADL) and 5alpha-pregnane-3alpha,20beta-diol (3alpha5alpha-diol) were significantly greater for the rat compared with the human receptor. Responses to 30 micromol/L GABA were inhibited by 3beta-OH-5alpha-pregnan-20-one (UC1010) and 5beta-pregnan-3beta,20(R)-diol (UC1020) to a greater degree for human and rat receptors, respectively. Responses to GABA + 3alpha5alphaTHDOC were inhibited by 5alpha-pregnan-3beta,20(S)-diol (UC1019) and pregnenolone sulphate to a greater degree for human and rat receptors, respectively. The GABA dose response curves for human alpha(1)beta(2)gamma(2)(S) and alpha(1)beta(2)gamma(2)(L) receptors were identical. However, the maximum GABA evoked current, the direct gating effect of pentobarbital and the allosteric potentiation of the GABA EC(15) response by 3alpha5alphaTHDOC and 3alpha5betaP were significantly higher with alpha(1)beta(2)gamma(2)(S) than alpha(1)beta(2)gamma(2)(L) receptors. Inhibition of the response to 30 micromol/L GABA by UC1010 and UC1020 was greater for a(1)beta(2)gamma(2)(L) and alpha(1)beta(2)gamma(2)(S) receptors, respectively. Inhibition of responses to 3alpha5alphaTHDOC + GABA by UC1019 and UC1010 was significantly higher for alpha(1)beta(2)gamma(2)(L) receptors. In conclusion, the site of activation by GABA and neurosteroid modulation differ between human and rat alpha(1)beta(2)gamma(2)(L) receptors, as well as between human receptors containing the L and S splice variants of the gamma(2)-subunit. PMID- 18430053 TI - Effects of atorvastatin and pravastatin on glucose tolerance, adipokine levels and inflammatory markers in hypercholesterolaemic patients. AB - Several randomized clinical trials have suggested that atorvastatin and pravastatin may differ in terms of their pleiotropic effects. To verify this, we compared the effects of both statins on glucose tolerance, adipokine concentrations and inflammatory markers. A total of 36 hypercholesterolaemic patients without known coronary heart disease (CHD) were enrolled in an open label, randomized, crossover study. The patients received pravastatin or atorvastatin (10 mg/day) for 4 months and then switched to the other statin for an additional 4 months. At the end of both treatment periods, atorvastatin significantly reduced the concentration of serum lipids (total and low-density lipoprotein-cholesterol and triglycerides) and inflammatory markers (high sensitivity C-reactive protein and tumour necrosis factor-a) and increased serum adiponectin levels compared with pravastatin treatment. Although these effects would be expected to improve insulin sensitivity, atorvastatin did not affect glucose tolerance, which was assessed by fasting glucose and insulin concentrations, the homeostasis model assessment index and glycosylated haemoglobin (HbA(1c)) levels. Only obese patients showed increased HbA(1c) levels after atorvastatin treatment. Our results suggest that atorvastatin has both advantages and disadvantages compared with pravastatin treatment. Further studies are required to compare the relative clinical value of atorvastatin and pravastatin, especially in obese patients without CHD. PMID- 18430054 TI - Tissue- and fibre-specific modifications of insulin-signalling molecules in cardiac and skeletal muscle of diabetic rats. AB - 1. Levels of insulin-signalling molecules are altered in streptozotocin (STZ) induced diabetes, a model of Type 1 diabetes. However, the tissue-specific regulation of these changes and the effect of insulin supplementation on signalling molecule protein levels have not been well characterized. 2. In the present study, we evaluated the level of proximal insulin-signalling intermediates in the heart and in red and white gastrocnemius muscles of 2 week diabetic rats and diabetic rats supplemented with insulin. 3. Diabetes augmented levels of the insulin receptor and the p85 regulatory subunit of phosphatidylinositol 3-kinase in the red gastrocnemius, but not in the white gastrocnemius or the heart. Furthermore, diabetes reduced insulin receptor substrate-1 levels in both the red and white gastrocnemius, but not in the heart. Examination of the levels and basal activities of distal insulin-signalling intermediates (protein kinase B (PKB)/Akt, extracellular signal-regulated kinase (ERK) 1/2, p38 mitogen-activated protein kinase (MAPK)) also failed to reveal a specific pattern in these changes. Thus, diabetes reduced basal ERK1/2 and PKB/Akt phosphorylation in the heart and white gastrocnemius, respectively, whereas it augmented basal p38 MAPK activity in the red gastrocnemius. Insulin supplementation normalized the levels and activities of some but not all proteins. 4. In conclusion, the results of the present study demonstrate that adaptation to STZ-induced diabetes varies among skeletal muscle fibre types and the heart, emphasizing the complex tissue-specific responses to diabetes. PMID- 18430055 TI - Identification of natural coumarin compounds that rescue defective DeltaF508-CFTR chloride channel gating. AB - 1. Deletion of phenylalanine at position 508 (DeltaF508) of the cystic fibrosis transmembrane conductance regulator (CFTR) chloride channel is the most common mutation causing cystic fibrosis (CF). Effective pharmacological therapy of CF caused by the DeltaF508-CFTR mutation requires the rescue of both intracellular processing and channel gating defects. 2. We identified a class of natural coumarin compounds that can correct the defective DeltaF508-CFTR chloride channel gating by screening a collection of 386 single natural compounds from Chinese medicinal herbs. Screening was performed with an iodide influx assay in Fischer rat thyroid epithelial cells coexpressing DeltaF508-CFTR and an iodide-sensitive fluorescent indicator (YFP-H148Q/I152L). 3. Dose-dependent potentiation of defective DeltaF508-CFTR chloride channel gating by five coumarin compounds was demonstrated by the fluorescent iodide influx assay and confirmed by an Ussing chamber short-circuit current assay. Activation was fully abolished by the specific CFTR inhibitor CFTR(inh)-172. Two potent compounds, namely imperatorin and osthole, have activation K(d) values of approximately 10 micromol/L, as determined by the short-circuit current assay. The active coumarin compounds do not elevate intracellular cAMP levels. Activation of DeltaF508-CFTR by the coumarin compounds requires cAMP agonist, suggesting direct interaction with the mutant CFTR molecule. Kinetics analysis indicated rapid activation of DeltaF508 CFTR by the coumarin compounds, with half-maximal activation of < 5 min. The activating effect was fully reversed for all five active compounds 45 min after washout. 4. In conclusion, the natural coumarin DeltaF508-CFTR activators may represent a new class of natural lead compounds for the development of pharmacological therapies for CF caused by the DeltaF508 mutation. PMID- 18430056 TI - Intraperitoneal glycerol induces oxidative stress in rat kidney. AB - 1. Glycerol has been used for the treatment of intracranial hypertension, cerebral oedema and glaucoma. Experimentally, intramuscular administration of hypertonic glycerol solution is used to produce acute renal failure. In this model, glycerol causes rhabdomyolysis and myoglobinuria, resulting in the development of renal injury. The pathogenesis is thought to involve vascular congestion, the formation of casts and oxidative stress. However, the effect of glycerol itself independent of rhabdomyolysis has not been investigated. Therefore, the aim of the present study was to investigate the effects of i.p. glycerol on some biochemical and oxidative stress parameters in the kidney of young rats. 2. Rats received 10 mL/kg, i.p., hypertonic glycerol solution (50% v/v) or saline (NaCl 0.85 g%) followed by 24 h water deprivation. Twenty-four hours after the administration of glycerol, rats were killed. Creatinine levels and the activity of creatine kinase (CK) and lactate dehydrogenase (LDH) were determined in the plasma. In addition, CK, pyruvate kinase and LDH activity and oxidative stress parameters (free radical formation, lipid peroxidation and protein carbonylation) were measured in renal tissue. 3. Glycerol did not alter plasma CK activity and increased plasma creatinine levels, suggesting renal insufficiency and the absence of rhabdomyolysis. Renal CK and pyruvate kinase activity was decreased, suggesting diminution of energy homeostasis in the kidney. Plasma and renal LDH activity was decreased, whereas the formation of free radicals, lipid peroxidation and protein carbonylation were increased, suggesting oxidative stress. 4. These results are similar to those described after the intramuscular administration of glycerol. Therefore, it is possible that glycerol may provoke renal lesions by mechanisms other than those induced by rhabdomyolysis. PMID- 18430057 TI - Effect of statin therapy on plasma high-density lipoprotein-cholesterol levels is modified by paraoxonase 1 in Chinese patients with coronary heart disease. AB - 1. We sought to determine the effects of Q192R polymorphism of paraoxonase 1 (PON1) gene on plasma high-density lipoprotein-cholesterol (HDL-C) levels and the response to statin therapy in Chinese patients with coronary heart disease (CHD). 2. Two hundred and thirty-six patients with CHD were treated with simvastatin 20 mg/day. Fasting serum lipids were determined before and after 12 weeks of treatment. 3. No significant differences were detected among the PON1 Q192R polymorphism with respect to plasma lipids. In addition, the effects of simvastatin to increase HDL-C levels were significantly greater in patients with the RR genotype compared with patients with the QR or RR genotypes (P < 0.05). 4. We conclude that the Q192R polymorphism of PON1 significantly modulates the HDL-C response to simvastatin in Chinese patients with CHD. PMID- 18430058 TI - Hyperhomocysteinaemia-induced cardiovascular changes in rats. AB - (1) Increased plasma homocysteine content and increased blood pressure are independently associated with higher cardiovascular risks. The present study was designed to determine the effects of hyperhomocysteinaemia (HHcys) on the activity of the cardiovascular system in rats. (2) Using male Wistar rats, the effect of moderate HHcys, induced by treating rats with dl-homocysteine thiolactone (DL-HT; 1 g/kg per day) for 15 days, on arterial blood pressure, heart rate, baroreflex and vascular reactivity was determined. (3) Hyperhomocysteinaemia was observed after 15 days of treatment. Baseline arterial blood pressure and heart rate values of HHcys animals were significantly increased after 15 days of treatment. Plasma homocysteine and cardiovascular parameters returned to control values after termination of treatment. Baroreflex gain was significantly enhanced in HHcys rats. The pressor effect of an i.v. infusion of phenylephrine (50 mg/kg per mL) was decreased in HHcys rats and returned to control values after washout of DL-HT. Hypotensive responses to i.v. infusions of sodium nitroprusside (70 mg/kg per mL) or acetylcholine (10 mg/kg per mL) were increased in HHcys animals and returned to control values after washout of DL-HT. The increase in resting arterial blood pressure associated with the moderate HHcys was reversed by treatment with the b1-adrenoceptor antagonist atenolol, suggesting that HHcys-related hypertension is related to increase in cardiac sympathetic activity. (4) The present study showed significantly increased arterial blood pressure, heart rate and baroreflex activity in the early phase of moderate HHcys. In addition, HHcys was associated with alterations of vascular responsiveness to pressor and depressor agents, as well as increased cardiac sympathetic activity. The fact that cardiovascular changes observed in HHcys were reversed after DL-HT washout indicate that moderate HHcys evokes cardiovascular changes. PMID- 18430059 TI - Effects of hesperidin on cyclic strain-induced endothelin-1 release in human umbilical vein endothelial cells. AB - 1. Hesperidin, a member of the flavanone group of flavonoids, can be isolated in large amounts from the rinds of some citrus species and has been reported to have antihypotensive and vasodilator properties. However, the mechanism of action of hesperidin in the prevention and treatment of vascular diseases remains unclear. 2. The vascular endothelium can produce potent contracting factors, such as endothelin (ET)-1, and endothelium-derived relaxing factors, such as nitric oxide (NO). The aims of the present study were to test the hypothesis that hesperidin may alter strain-induced ET-1 secretion and NO production and to identify the putative underlying signalling pathways in human umbilical vein endothelial cells (HUVEC). 3. Hesperidin (10 and 100 micromol/L) inhibited strain-induced ET-1 secretion. Hesperidin also inhibited strain-induced increases in the formation of reactive oxygen species and extracellular signal-regulated kinase (ERK) phosphorylation. 4. Hesperidin treatment of HUVEC enhanced NO production, endothelial NO synthase (eNOS) activity and the phosphorylation of eNOS and Akt. Furthermore, hesperidin modulated strain-induced ET-1 release and suppressed ERK phosphorylation in part via the NO/protein kinase G pathway. 5. In summary, we have demonstrated that hesperidin inhibits strain-induced ET-1 secretion and enhances NO production in HUVEC. PMID- 18430061 TI - Elevated catalase and heme oxygenase-1 may contribute to improved postischaemic cardiac function in long-term type 1 diabetes. AB - 1. Although increased oxidative stress has been shown repeatedly to be implicated in diabetes, the cardiovascular anti-oxidant state and heart response to ischaemia in long-term Type 1 diabetes remain largely unknown. The present study was designed to observe heart tolerance to ischaemia-reperfusion and endogenous anti-oxidants in the cardiovascular system in long-term hyperglycaemic rats. 2. Hearts from Sprague-Dawley rats surviving up to 6 months with streptozocin induced severe hyperglycaemia (blood glucose > 20 mmol/L) were isolated and subjected to global ischaemia and reperfusion. Cardiac function, electrocardiogram and anti-oxidants in the myocardium and aorta were examined. In addition, the morphology of the myocardial mitochondria and the in vitro function of aortic vessels were assessed. 3. Hearts from diabetic rats demonstrated lower baseline heart function but had higher postischaemic coronary flow and left ventricular developed pressure compared with their respective controls (P < 0.05). In addition, hearts from diabetic animals had fewer arrhythmias (P < 0.01) and lower left ventricular end-diastolic pressure during reperfusion (P < 0.05). Higher catalase and heme oxygenase-1 content was found in the aorta and myocardium from diabetic rats (P < 0.01). In aortas from diabetic animals, acetylcholine-induced vasodilatation was enhanced and was approximately 15% after inhibition of nitric oxide synthase, compared with 0% in controls. The 15% relaxation was abrogated by heme oxygenase blockade. Mitochondria from the myocardium of diabetic rats showed significant increases in both size and number (P < 0.05). 4. Hearts of long-term Type 1 diabetic rats demonstrated improved recovery of postischaemic cardiac function and reduced reperfusion arrhythmia. Hyperglycaemia may enhance cardiovascular anti-oxidant capacity and mitochondrial neogenesis, which renders the heart resistant to ischaemia and oxidative injury. PMID- 18430060 TI - Sequential activation of the reactive oxygen species/angiotensinogen/renin angiotensin system axis in renal injury of type 2 diabetic rats. AB - 1. The present study was performed to test the hypothesis that the reactive oxygen species (ROS)-angiotensinogen (AGT)-renin angiotensin system (RAS) axis is sequentially activated in the development of diabetic nephropathy in Zucker diabetic fatty (ZDF) obese rats. 2. Genetic pairs of male ZDF obese and control ZDF lean rats (n = 12 of each species) were killed every 3 weeks from 12 to 21 weeks of age (n = 6 at each time point). 3. The ZDF obese rats developed diabetes mellitus at 12 weeks. At that time, urinary excretion rates of 8-isoprostane were similar between the groups; however, urinary 8-isoprostane levels were significantly increased at 15 weeks in ZDF obese rats compared with controls (36 +/- 6 vs 15 +/- 2 ng/day, respectively). At 15 weeks, protein levels of cortical angiotensinogen were similar between groups; however, cortical angiotensinogen levels were significantly increased at 18 weeks in ZDF obese rats compared with controls (relative ratio of 2.32 +/- 0.21 vs 1.00 +/- 0.20, respectively). At 12 weeks, angiotensin (Ang) II-like immunoreactivity was similar between groups in both the glomeruli and tubules; however, AngII-like immunoreactivity was increased significantly at 21 weeks in ZDF obese rats compared with controls (relative ratios of 1.98 +/- 0.55 vs 1.00 +/- 0.03, respectively, for glomeruli and 1.58 +/- 0.16 vs 1.00 +/- 0.13, respectively, for tubules). Moreover, at 21 weeks, the desmin-positive area in the glomeruli (0.63 +/- 0.08 vs 0.22 +/- 0.05%) and Masson's trichrome stain-positive area in the interstitium (4.97 +/- 0.05 vs 3.18 +/- 0.41%) were significantly increased in ZDF obese rats compared with controls, even though these differences had not been observed earlier. 4. These data suggest that the sequential activation of the ROS-AGT-RAS axis plays an important role in the development of diabetic nephropathy in ZDF obese rats. PMID- 18430062 TI - Effects of a chronic exercise training protocol on oxidative stress and right ventricular hypertrophy in monocrotaline-treated rats. AB - 1. In the present study, we investigated the effects of exercise training on cardiac hypertrophy and oxidative stress in a monocrotaline (MCT)-induced cor pulmonale model. Male Wistar rats were assigned to one of three groups: sedentary control (SC); sedentary MCT (SM); or trained MCT (TM). 2. Right ventricular hypertrophy (RVH) was induced by a single injection of MCT (60 mg/kg, i.p.). Exercise training consisted of running on a treadmill (five times a week, during Weeks 3, 4 and 5). Systemic oxidative stress was evaluated in erythrocytes by chemiluminescence (CL) and the activity of the anti-oxidant enzymes superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and glutathione S transferase (GST) was determined. 3. At Week 3, MCT-treated animals exhibited RVH, lung congestion, decreased SOD activity and increased CAT activity. Exercise training reduced MCT-induced RVH and increased GST activity. At Week 4, MCT induced RVH was accompanied by an increase in CL and GST activity. However, in TM animals there was a decrease in CL and augmented SOD activity. At Week 5, there were no survivors in the SM group, whereas GST activity was elevated in TM rats compared with SC rats. There was no difference in GPx activity throughout the experimental protocol between the groups. 4. Taken together, our results indicate that exercise training is able to ameliorate RVH and improve survival, which is associated with a reduction in oxidative stress in MCT-treated rats. PMID- 18430063 TI - Antiproliferative activity of aucubin is through cell cycle arrest and apoptosis in human non-small cell lung cancer A549 cells. AB - Aucubin, an iridoid glycoside isolated from the leaves of Aucuba japonica, inhibits human non-small cell lung cancer A549 cells by blocking cell cycle progression in the G(0)/G(1) phase and inducing apoptosis. An ELISA showed that the G(0)/G(1) phase arrest is due to p53-mediated induction of p21. Enhancement of Fas and its two ligands, membrane-bound and soluble Fas ligand, may be responsible for the apoptotic effect induced by aucubin. The present study shows, for the first time, that the induction of p53 and activity of the Fas/Fas ligand apoptotic system may participate in the antiproliferative activity of aucubin in A549 cells. PMID- 18430066 TI - Indicators of dehydration and haemoconcentration are associated with the prevalence and severity of coronary artery disease. AB - 1. The vascular endothelium is injured by blood flow abnormalities exacerbated by different risk factors, including markers of haemoconcentration. The aim of the present study was to assess the association between markers of haemoconcentration and dehydration and the prevalence and severity of coronary artery disease (CAD). 2. Subjects in the present study (189 men and 126 women) were classified as either CAD cases or controls according to the results of coronary angiography. The severity of CAD was scored on the basis of the number and the extent of lesions on coronary arteries. Serum electrolytes, osmolality and haematological parameters were measured. 3. Compared with control subjects, patient with CAD had increased levels of serum osmolality, calculated osmolality, tonicity, sodium, glucose and blood urea nitrogen (BUN). Significant differences were also observed in the haematocrit and haemoglobin concentration, but not in erythrocyte counts and total serum protein. On multiple logistic regression analysis adjusting for major risk factors, serum osmolality, glucose and BUN exhibited significant associations with CAD, but the correlations were lessened by diabetes. Analysis using anova showed a significant correlation between serum osmolality, sodium, glucose and BUN and the severity of CAD. The area under the receiver operating characteristic curves, as a relative measure of the test's efficiency, was the highest and significant for serum osmolality, BUN and glucose. 4. The results indicate that some of the markers of dehydration and haemoconcentration are associated significantly with the prevalence and severity of CAD, but the independence of these correlations is questioned. These markers may play a role in the pathogenesis of atherosclerosis. PMID- 18430065 TI - Both 5-hydroxytryptamine 5-HT2A and 5-HT1B receptors are involved in the vasoconstrictor response to 5-HT in the human isolated internal thoracic artery. AB - 1. The 5-hydroxytryptamine (5-HT, serotonin) receptor subtypes that mediate vasoconstriction in the human internal thoracic artery (ITA), which is frequently used as an arterial graft, remain unclear. The aim of the present study was to elucidate the 5-HT receptor subtypes responsible for 5-HT-induced contraction of the human ITA. 2. The contractile responses to 5-HT of endothelium-denuded human ITA obtained from patients undergoing coronary bypass surgery were examined. In addition, we investigated the effects of sarpogrelate and SB224289, antagonists of 5-HT(2A) and 5-HT(1B) receptors, respectively, on the 5-HT-induced vasoconstriction. Finally, 5-HT(2A) and 5-HT(1B) receptors in the human ITA were immunolabelled. 3. 5-Hydroxytryptamine (1 nmol/L-10 micromol/L) caused vasoconstriction in a concentration-dependent manner. Both sarpogrelate (1 micromol/L) and SB224289 (1 micromol/L) significantly, but not completely, inhibited 5-HT-induced vasoconstriction. 4. Conversely, simultaneous pretreatment with supramaximum concentrations (1 micromol/L for both) of sarpogrelate and SB224289 almost completely inhibited the 5-HT-induced vasoconstriction. 5. Immunopositive staining for 5-HT(2A) and 5-HT(1B) receptors was detected in smooth muscle cells of the human ITA. 6. These results demonstrate that, in human ITA, 5-HT-induced vasoconstriction is mediated by activation of both 5-HT(2A) and 5-HT(1B) receptors. Thus, when the human ITA is used as an arterial graft, a combination of 5-HT(2A) and 5-HT(1B) receptor antagonists would appear to be most useful to prevent 5-HT-induced vasospasm. PMID- 18430064 TI - Topical oxygen therapy induces vascular endothelial growth factor expression and improves closure of clinically presented chronic wounds. AB - 1. Chronic wounds, especially in diabetics, represent a serious threat to human health. 2. Correcting a compromised state of tissue oxygenation by the administration of supplemental O(2) is known to benefit wound healing. Beyond its role as a nutrient and antibiotic, O(2) supports wound healing by driving redox signaling. 3. Hyperbaric oxygen (HBO) therapy is widely used and approved by Center for Medicare and Medicaid Services to treat specific ulcerations. The current literature supports the notion that approaches to topically oxygenate wounds may be productive. 4. Here, we present the results of two simultaneous studies testing the effects of HBO and portable topical oxygen (TO) therapies. These two therapeutic approaches have several contrasting features. 5. In total, 1854 patients were screened in outpatient wound clinics for non-randomized enrolments into the HBO (n = 32; 31% diabetic) and TO (n = 25; 52% diabetic) studies. 6. Under the conditions of the present study, HBO treatment seemed to benefit some wounds while not benefiting others. Overall, HBO did not result in statistically significant improvements in wound size in the given population over the time monitored in the present study. 7. However, TO significantly improved wound size. Among the three O(2)-sensitive genes (VEGF, TGFbeta1 and COL1A1) studied in wound edge tissue biopsies, TO treatment was associated with higher VEGF165 expression in healing wounds. Expression of the other genes mentioned was not affected by TO. There was no significant change in the expression levels of any of genes studied in patients in the HBO study. This establishes a link between VEGF gene expression and healing outcome for TO therapy. 8. Taken together, the present study provides evidence demonstrating that TO treatment benefits wound healing in patients suffering from chronic wounds. Treatment with TO is associated with an induction of VEGF expression in wound edge tissue and an improvement in wound size. PMID- 18430068 TI - Guidelines for diagnosing and treating pulmonary infiltrates in children with acute leukaemia: impact of timely decisions. AB - AIM: Children with leukaemia are at increased risk of pulmonary complications, often with unspecific clinical data, delayed diagnosis and a high mortality rate. We evaluated the usefulness of diagnostic-therapeutic guidelines (DTG) in which specific times for decision making were incorporated. METHODS: Clinical charts of children with acute leukaemia and suspicion of pulmonary involvement were reviewed. Patients were allocated to group I if their diagnostic and therapeutic decisions were in accordance with the DTG, and to group II if not. RESULTS: Children from group I (n=32) and group II (n=28) did not differ with respect to age (9.3+/-0.5 years old, mean+/-SEM), gender, type, risk and stage of leukaemia, anaemia and neutropenia. Total length of hospital stay and hospitalization due to the pulmonary disease were shorter in group I than in group II (14.8+/-2.1 vs. 28.5+/-3.7 days, p=0.0016; and 10.8+/-1.0 vs. 18.4+/-1.8 days, p=0.0003, respectively). Two patients (6.3%) died due to the pulmonary pathology in group I, and nine (32.1%, p=0.016) in group II. CONCLUSIONS: Diagnostic-therapeutic guidelines that incorporate timely decisions constitute a useful algorithm to reduce the length of hospital stay and mortality in children with acute leukaemia and pulmonary infiltrates. A prospective study is needed to validate these results. PMID- 18430067 TI - The epithelial brush border Na+/H+ exchanger NHE3 associates with the actin cytoskeleton by binding to ezrin directly and via PDZ domain-containing Na+/H+ exchanger regulatory factor (NHERF) proteins. AB - 1. The Na(+)/H(+) exchanger NHE3 associates with the actin cytoskeleton by binding ezrin both directly and indirectly. Both types of interaction are necessary for acute regulation of NHE3. Most acute regulation of NHE3 occurs by changes in trafficking via effects on exocytosis and/or endocytosis. However, NHE3 activity can also be regulated without changing the surface expression of NHE3 (change in turnover number). 2. A positive amino acid cluster in the a helical juxtamembrane region in the COOH-terminus of NHE3 (amino acids K516, R520 and R527) is necessary for binding to the protein 4.1, ezrin, radixin, moesin (FERM) domain III of ezrin. Direct binding of NHE3 to ezrin is necessary for many aspects of basal trafficking, including basal exocytosis, delivery from the synthetic pathway and movement of NHE3 in the brush border (BB), which probably contributes to endocytosis over a prolonged period of time. 3. In addition, NHE3 binds indirectly to ezrin. The PDZ domain-containing proteins Na(+)/H(+) exchanger regulatory factor (NHERF) 1 and NHERF2, as intermediates in linking NHE3 to ezrin, are necessary for many aspects of NHE3 regulation. The binding of NHERF-ezrin/radixin/moesin to NHE3 occurs in the cytosolic domain of NHE3 between amino acids 475 and 689. This NHERF binding is involved in the formation of the NHE3 complex and restricts NHE3 mobility in the BB. However, it is dynamic; for example, changing in some cases of signalling. Furthermore, NHERF binding is necessary for lysophosphatidic acid stimulation of NHE3 and inhibition of NHE3 by Ca(2+), cAMP and cGMP. PMID- 18430069 TI - Endothelial function in children with a history of premature prolonged rupture of membranes and bronchopulmonary dysplasia--a pilot study. AB - AIM: To investigate vascular function in children with a neonatal history of generalised inflammation indicated by premature prolonged rupture of membranes (PPROM) and bronchopulmonary dysplasia (BPD). METHODS: Children born at 2 SD above the norm. Bioelectrical impedance analysis (BIA), measured with a multiple frequency analyzer, was used to estimate body composition. DNA extraction from white blood cells was used to estimate the telomere length by detection of terminal restriction fragments (TRF). RESULTS: No difference was found between the TRF lengths of obese and normal children. Obese adults had shorter TRF lengths than adults who were not obese (mean TRF length difference, 884.5; 95% confidence intervals -1727 to -41.8; t=2.183; df=17; p<0.041). CONCLUSIONS: Obese adults have shorter telomeres than their normal-weight counterparts, while this phenomenon is not present in childhood. PMID- 18430075 TI - Paediatric obesity: a neurodevelopmental perspective. PMID- 18430076 TI - Professionalism and psychiatry: the profession speaks. AB - OBJECTIVE: Recently in the USA and the UK, the medical profession as a whole and psychiatry in particular have felt 'deprofessionalised'. This study aimed to explore ideas about professionalism and the threat of deprofessionalisation from psychiatrists themselves. METHOD: Attendees at three meetings related to the activities of the Royal College of Psychiatrists were invited to respond to a short questionnaire. These were meetings on different days and different people attended these. The questionnaire had only five simple questions. In two meetings, all attendees answered and in the third meeting only two of 20 attendees did so. RESULTS: Seventy-three completed questionnaires were received. Of these, a vast majority, 89% of 66, believed that professionalism is important in modern day clinical practice; key threats to professionalism are the government, the media and other specialties and disciplines. There was broad agreement that the Royal College of Psychiatrists should lead and define professionalism. CONCLUSION: The ownership of professionalism by the professionals themselves is the most important factor; ethics, standards, attitudes and skills are seen as significant components of professionalism and should be taught early to trainees. PMID- 18430077 TI - Relationship between body mass index and weight perception in schizophrenia. PMID- 18430078 TI - Regulation of mycothiol metabolism by sigma(R) and the thiol redox sensor anti sigma factor RsrA. AB - Mycothiol (MSH) is the major thiol in Actinobacteria and plays a role analogous to that of glutathione. The biosynthetic pathway has been established in mycobacteria and is initiated by the glycosyltransferase MshA. A key mycothiol dependent detoxification pathway utilizes the amidase (Mca) to cleave mycothiol S conjugates to produce GlcN-Ins and a mercapturic acid excreted from the cell. How expression of mycothiol genes is regulated in mycobacteria has been unclear so the report in this issue by Park and Roe showing that in Streptomyces coelicolor the redox controlled anti-sigma factor RsrA that binds the regulator sigma(R) controls key elements of mycothiol metabolism is a major advance. Conditions that deplete thiols are shown to induce directly expression of sigR, rsrA, mshA and mca, as well as the thioredoxin reductase-thioredoxin system, generating an autoregulatory cycle that persists until the thiol-depleting condition is alleviated. Evidence for indirect induction of mshB-D to support mycothiol biosynthesis is also presented. It was shown in vitro that mycothiol, like reduced thioredoxin and dithiothreitol, can reduce oxidized RsrA to activate its binding to sigma(R). These studies establish for the first time how mycothiol metabolism is regulated to cope with stress from thiol reactive toxins. PMID- 18430079 TI - The 'core' and 'accessory' regulons of Pseudomonas-specific extracytoplasmic sigma factors. AB - Pyoverdine is a fluorescent, high-affinity peptide siderophore produced by different Pseudomonas species. The genes for pyoverdine biosynthesis depend on PvdS, an extracytoplasmic sigma factor. In this issue of Molecular Microbiology, Swingle et al. demonstrate that in the plant pathogen Pseudomonas syringae PvdS not only regulates the production of pyoverdine (core regulon), but also controls expression of other genes likely to be involved in the adaptation to the environment (accessory regulon). This accessory regulon is variable, as different sets of genes seem to be recruited according to the Pseudomonas species and its specific ecological niche. PMID- 18430080 TI - LysM, a widely distributed protein motif for binding to (peptido)glycans. AB - Bacteria retain certain proteins at their cell envelopes by attaching them in a non-covalent manner to peptidoglycan, using specific protein domains, such as the prominent LysM (Lysin Motif) domain. More than 4000 (Pfam PF01476) proteins of both prokaryotes and eukaryotes have been found to contain one or more Lysin Motifs. Notably, this collection contains not only truly secreted proteins, but also (outer-)membrane proteins, lipoproteins or proteins bound to the cell wall in a (non-)covalent manner. The motif typically ranges in length from 44 to 65 amino acid residues and binds to various types of peptidoglycan and chitin, most likely recognizing the N-acetylglucosamine moiety. Most bacterial LysM-containing proteins are peptidoglycan hydrolases with various cleavage specificities. Binding of certain LysM proteins to cells of Gram-positive bacteria has been shown to occur at specific sites, as binding elsewhere is hindered by the presence of other cell wall components such as lipoteichoic acids. Interestingly, LysM domains of certain plant kinases enable the plant to recognize its symbiotic bacteria or sense and induce resistance against fungi. This interaction is triggered by chitin-like compounds that are secreted by the symbiotic bacteria or released from fungi, demonstrating an important sensing function of LysMs. PMID- 18430081 TI - PerR acts as a switch for oxygen tolerance in the strict anaerobe Clostridium acetobutylicum. AB - Clostridia belong to those bacteria which are considered as obligate anaerobe, e.g. oxygen is harmful or lethal to these bacteria. Nevertheless, it is known that they can survive limited exposure to air, and often eliminate oxygen or reactive derivatives via NAD(P)H-dependent reduction. This system does apparently contribute to survival after oxidative stress, but is insufficient to establish long-term tolerance of aerobic conditions. Here we show that manipulation of the regulatory mechanism of this defence mechanism can trigger aerotolerance in the obligate anaerobe Clostridium acetobutylicum. Deletion of a peroxide repressor (PerR)-homologous protein resulted in prolonged aerotolerance, limited growth under aerobic conditions and rapid consumption of oxygen from an aerobic environment. The mutant strain also revealed higher resistance to H2O2 and activities of NADH-dependent scavenging of H2O2 and organic peroxides in cell free extracts increased by at least one order of magnitude. Several genes encoding the putative enzymes were upregulated and identified as members of the clostridial PerR regulon, including the heat shock protein Hsp21, a reverse rubrerythrin which was massively produced and became the most abundant protein in the absence of PerR. This multifunctional protein is proposed to play the crucial role in the oxidative stress defence. PMID- 18430082 TI - Mycothiol regulates and is regulated by a thiol-specific antisigma factor RsrA and sigma(R) in Streptomyces coelicolor. AB - Mycothiol (MSH) is a small thiol molecule with a cysteine-ligated disaccharide structure found in actinomycetes that include streptomycetes and mycobacteria. In Streptomyces coelicolor, a model organism for antibiotic production and differentiation, the amount of MSH is under the control of a sigma factor sigma(R), which is regulated by an antisigma factor RsrA with a thiol-disulphide redox switch. We found that the first gene (mshA) in the biosynthetic pathway for MSH and the gene for amidase (mca) that participates in detoxifying mycothiol reactive drugs are under direct control of sigma(R). The sigma(R) target genes are induced not only by a thiol oxidant diamide, but also by alkylating agents that cause a rapid decrease in MSH. Expression of the sigma(R) regulon was also elevated in MSH-deficient mutants, suggesting that a decrease in the level of MSH is a natural intracellular trigger for sigma(R) activation. We found that MSH was capable of reducing RsrA to bind sigma(R), whereas glutathione was not. These results support a proposal that the RsrA-sigma(R) system senses the intracellular level of reduced MSH, and that MSH serves as a natural modulator of the transcription system for its own replenishment in addition to being a redox buffer and drug detoxifier. PMID- 18430083 TI - The LAMP-like protein p67 plays an essential role in the lysosome of African trypanosomes. AB - RNAi knockdown was employed to study the function of p67, a lysosome-associated membrane protein (LAMP)-like type I transmembrane lysosomal glycoprotein in African trypanosomes. Conditional induction of p67 dsRNA resulted in specific approximately 90% reductions in de novo p67 synthesis in both mammalian bloodstream and procyclic insect-stage parasites. Bloodstream cell growth was severely retarded with extensive death after > 24 h of induction. Biosynthetic trafficking of residual p67, and of the soluble lysosomal protease trypanopain, were unimpaired. Endocytosis of tomato lectin, a surrogate receptor-mediated cargo, was only mildly impaired (approximately 20%), but proper lysosomal targeting was unaffected. p67 ablation had dramatic effects on lysosomal morphology with gross enlargement (four- to fivefold) and internal membrane profiles reminiscent of autophagic vacuoles. Ablation of p67 expression rendered bloodstream trypanosomes refractory to lysis by human trypanolytic factor (TLF), a lysosomally activated host innate immune mediator. Similar effects on lysosomal morphology and TLF sensitivity were also obtained by two pharmacological agents that neutralize lysosomal pH--chloroquine and bafilomycin A1. Surprisingly, however, lysosomal pH was not affected in ablated cells suggesting that other physiological alterations must account for increased resistance to TLF. These results indicate p67 plays an essential role in maintenance of normal lysosomal structure and physiology in bloodstream-stage African trypanosomes. PMID- 18430084 TI - A novel autotransporter of uropathogenic Proteus mirabilis is both a cytotoxin and an agglutinin. AB - One of the six predicted Proteus mirabilis autotransporters (ATs), ORF c2341, is predicted to contain a serine protease motif and was earlier identified as an immunogenic outer membrane protein in P. mirabilis. The 3.2 kb gene encodes a 117 kDa protein with a 58-amino-acid-long signal peptide, a 75-kDa-long N-terminal passenger domain and a 30-kDa-long C-terminal translocator. Affinity-purified 110 kDa AT exhibited chymotrypsin-like activity and hydrolysed N-Suc-Ala-Ala-Pro-Phe pNa and N-Suc-Ala-Ala-Pro-Leu-pNa with a K(M) of 22 muM and 31 muM, respectively, under optimal pH of 8.5-9.0 in a Ca(2+)-dependent manner. Activity was inhibited by subtilase-specific inhibitors leupeptin and chymostatin. Both the cell associated and purified form elicited cytopathic effects on cultured kidney and bladder epithelial cells. Substrate hydrolysis as well as cytotoxicity was associated with the passenger domain and was compromised upon mutation of any of the catalytic residues (Ser366, His147 and Asp533). At alkaline pH and optimal cell density, the AT also promoted autoaggregation of P. mirabilis and this function was independent of its protease activity. Cytotoxicity, autoaggregation and virulence were significantly reduced in an isogenic pta mutant of P. mirabilis. Proteus toxic agglutinin (Pta) represents a novel autotransported cytotoxin with no bacterial homologues that works optimally in the alkalinized urinary tract, a characteristic of urease-mediated urea hydrolysis during P. mirabilis infection. PMID- 18430085 TI - Loss of tafazzin in yeast leads to increased oxidative stress during respiratory growth. AB - The tafazzin (TAZ) gene is highly conserved from yeast to humans, and the yeast taz1 null mutant shows alterations in cardiolipin (CL) metabolism, mitochondrial dysfunction and stabilization of supercomplexes similar to those found in Barth syndrome, a human disorder resulting from loss of tafazzin. We have previously shown that the yeast tafazzin mutant taz1Delta, which cannot remodel CL, is ethanol-sensitive at elevated temperature. In the current report, we show that in response to ethanol, CL mutants taz1Delta as well as crd1Delta, which cannot synthesize CL, exhibited increased protein carbonylation, an indicator of reactive oxygen species (ROS). The increase in ROS is most likely not due to defective oxidant defence systems, as the CL mutants do not display sensitivity to paraquat, menadione or hydrogen peroxide (H2O2). Ethanol sensitivity and increased protein carbonylation in the taz1Delta mutant but not in crd1Delta can be rescued by supplementation with oleic acid, suggesting that oleoyl-CL and/or oleoyl-monolyso-CL enables growth of taz1Delta in ethanol by decreasing oxidative stress. Our findings of increased oxidative stress in the taz1Delta mutant during respiratory growth may have important implications for understanding the pathogenesis of Barth syndrome. PMID- 18430086 TI - Neurogenic development of the visual areas in the Chinese softshell turtle (Pelodiscus sinensis) and evolutionary implications. AB - To characterize the neurogenic development of the visual areas of the turtle (Pelodiscus sinensis) during embryogenesis, a single dose of [(3)H]-thymidine (10 microCi) was injected into egg yolks from stages S11-12 to S21. At hatching, localization of [(3)H]-thymidine incorporation was examined, and led to three main observations. (1) Neurogenesis occurred in the stratum griseum centrale of the tectum opticum from S11-12 to S16 with a peak at S12. No obvious gradients of neurogenesis were observed. (2) Neurogenesis in the nucleus rotundus (Rot) and in the dorsal lateral geniculate nucleus (GLd) occurred from S11-12 to S15. Gradients of neurogenesis were detected along ventral-dorsal and lateral-medial axes in the Rot, but only the latter neurogenic gradient occurred in the GLd. (3) In the visual region of the dorsal ventricular ridge, neurogenesis lasted from S11-12 to S16. Similarly, neurogenesis occurred from S11-12 to S16-17 in the dorsal cortex, with a peak at S12 for both telencephalic visual regions. Neurogenesis followed a ventrolateral to dorsomedial gradient in the visual region of the dorsal ventricular ridge, and a superficial to deep gradient in the caudal dorsal cortex. A significant number of neurons in the rostral dorsal cortex followed a deep (earlier arising) to superficial (later arising) pattern of neurogenesis, similar to that in the avian Wulst or in the mammalian isocortex. Finally, we compared the timing and development of neurogenesis in the turtle with birds and mammals to understand the evolutionary implications of these processes. PMID- 18430087 TI - Visualizing the lateral somitic frontier in the Prx1Cre transgenic mouse. AB - Changes in the organization of the musculoskeletal system have accounted for many evolutionary adaptations in the vertebrate body plan. The musculoskeletal system develops from two mesodermal populations: somitic mesoderm gives rise to the axial skeleton and all of the skeletal muscle of the body, and lateral plate mesoderm gives rise to the appendicular skeleton. The recognition of embryonic domains resulting from the dynamics of morphogenesis has inspired new terminology based on developmental criteria. Two mesodermal domains are defined, primaxial and abaxial. The primaxial domain includes musculoskeletal structures comprising just somitic cells. The abaxial domain contains somitic myoblasts in connective tissue derived from lateral plate mesoderm, as well as lateral plate-derived skeletal structures. The boundary between these two domains is the lateral somitic frontier. Recent studies have described the developmental relationship between these two domains in the chick. In the present study, we describe the labelling pattern in the body of the Prx1/Cre/Z/AP compound transgenic mouse. The enhancer employed in this transgenic leads to reporter expression in the postcranial, somatic lateral plate mesoderm. The boundary between labelled and unlabelled cell populations is described at embryonic day (E)13.5 and E15.5. We argue that the distribution of labelled cells is consistent with the somatic lateral plate lineage, and therefore provides an estimate of the position of the lateral somitic frontier. The role of the frontier in both development and evolution is discussed. PMID- 18430088 TI - The lens-regenerating competence in the outer cornea and epidermis of larval Xenopus laevis is related to pax6 expression. AB - After lentectomy, larval Xenopus laevis can regenerate a new lens by transdifferentiation of the outer cornea and pericorneal epidermis (lentogenic area). This process is promoted by retinal factor(s) accumulated into the vitreous chamber. To understand the molecular basis of the lens-regenerating competence (i.e. the capacity to respond to the retinal factor forming a new lens) in the outer cornea and epidermis, we analysed the expression of otx2, pax6, sox3, pitx3, prox1, betaB1-cry (genes all involved in lens development) by Real-time RT-PCR in the cornea and epidermis fragments dissected from donor larvae. The same fragments were also implanted into the vitreous chamber of host larvae to ascertain their lens-regenerating competence using specific anti-lens antibodies. The results demonstrate that there is a tight correlation between lens-regenerating competence and pax6 expression. In fact, (1) pax6 is the only one of the aforesaid genes to be expressed in the lentogenic area; (2) pax6 expression is absent in head epidermis outside the lentogenic area and in flank epidermis, both incapable of transdifferentiating into lens after implantation into the vitreous chamber; (3) in larvae that have undergone eye transplantation under the head or flank epidermis, pax6 re-expression was observed only in the head epidermis covering the transplanted eye. This is consistent with the fact that only the head epidermis reacquires the lens-regenerating competence after eye transplantation, forming a lens following implantation into the vitreous chamber; and (4) in larvae that have undergone removal of the eye, the epidermis covering the orbit maintained pax6 expression. This is consistent with the fact that after the eye enucleation the lentogenic area maintains the lens regenerating competence, giving rise to a lens after implantation into the vitreous chamber. Moreover, we observed that misexpression of pax6 is sufficient to promote the acquisition of the lens-regenerating competence in flank epidermis. In fact, flank epidermis fragments dissected from pax6 RNA injected embryos could form lenses when implanted into the vitreous chamber. The data indicate for the first time that pax6 is a pivotal factor of lens-regenerating competence in the outer cornea and epidermis of larval X. laevis. PMID- 18430089 TI - The optic vesicle promotes cornea to lens transdifferentiation in larval Xenopus laevis. AB - The outer cornea and pericorneal epidermis (lentogenic area) of larval Xenopus laevis are the only epidermal regions competent to regenerate a lens under the influence of the retinal inducer. However, the head epidermis of the lentogenic area can acquire the lens-regenerating competence following transplantation of an eye beneath it. In this paper we demonstrate that both the outer cornea and the head epidermis covering a transplanted eye are capable of responding not only to the retinal inducer of the larval eye but also to the inductive action of the embryonic optic vesicle by synthesizing crystallins. As the optic vesicle is a very weak lens inductor, which promotes crystallin synthesis only on the lens biased ectoderm of the embryo, these results indicate that the lens-forming competence in the outer cornea and epidermis of larval X. laevis corresponds to the persistence and acquisition of a condition similar to that of the embryonic biased ectoderm. PMID- 18430090 TI - Three-dimensional ontogenetic shape changes in the human cranium during the fetal period. AB - Knowledge of the pattern of human craniofacial development in the fetal period is important for understanding the mechanisms underlying the emergence of variations in human craniofacial morphology. However, the precise character of the prenatal ontogenetic development of the human cranium has yet to be fully established. This study investigates ontogenetic changes in cranial shape in the fetal period, as exhibited in Japanese fetal specimens housed at Kyoto University. A total of 31 human fetal specimens aged from approximately 8 to 42 weeks of gestation underwent helical computed tomographic scanning, and 68 landmarks were digitized on the internal and external surfaces of the extracted crania. Ontogenetic shape change was then analyzed cross-sectionally and three-dimensionally using a geometric morphometric technique. The results of the present study are generally consistent with previously reported findings. It was found that during the prenatal ontogenetic process, the growth rate of the length of the cranium is greater than that of the width and height, and the growth rate of the length of the posterior cranial base is smaller than that of the anterior cranial base. Furthermore, it was observed that the change in shape of the human viscerocranium is smaller than that of the neurocranium during the fetal period, and that concurrently the basicranium extends by approximately 8 degrees due to the relative elevation of the basilar and lateral parts of occipital bone. These specific growth-related changes are the opposite of those reported for the postnatal period. Our findings therefore indicate that the allometric pattern of the human cranium is not a simple continuous transformation, but changes drastically from before to after birth. PMID- 18430091 TI - Anatomical, immnunohistochemical and physiological characteristics of the vomeronasal vessels in cows and their possible role in vomeronasal reception. AB - The general morphology of the vomeronasal vessels in adult cows was studied following a classic protocol, including optical, confocal and ultrastructural approaches. This anatomical work was completed immunohistochemically. The vomeronasal organ in cows is well developed, and its vessels are considerable in size. This fact allowed some functional properties of the vomeronasal arteries to be evaluated and, for the first time, their isometric tension to be recorded. Our functional studies were in agreement with the immunohistochemistry, and both corroborated the morphological data on the similarity between the vomeronasal vessels and those of the typical erectile tissue. In consequence, the vasoconstriction and vasodilation of the vomeronasal vessels would facilitate an influx and outflow of fluids in the vomeronasal organ, that is to say, this organ in cows would be able to work as a pump mechanism to send chemical signals to the vomeronasal receptor neurones. PMID- 18430094 TI - Erratum. PMID- 18430093 TI - Phase II study of hepatic arterial infusion of a fine-powder formulation of cisplatin for advanced hepatocellular carcinoma. AB - AIM: Intra-arterial cisplatin appears to have high therapeutic efficacy, but this has not been studied in detail. Accordingly, we developed a fine-powder cisplatin formulation and tested it in advanced hepatocellular carcinoma (HCC) patients in an open-label, uncontrolled study in which 27 institutions participated. METHODS: The patients in this study had inoperable advanced HCC without extrahepatic metastases. All received two infusions of high-concentration cisplatin (1.43 mg/mL) via the hepatic artery at a dose of 65 mg/m(2), with an intervening 4-6 week interval. RESULTS: Cisplatin efficacy and safety were assessed in 80 patients. We found partial responses in 27 cases, no change in 37, and progressive disease in 11 (five were not evaluated). The overall response rate was 33.8%. The 1-year survival rate was 67.5% and the 2-year survival rate was 50.8%. Severe adverse effects (>/=grade 3) included anorexia in 22.5%, vomiting in 6.3%, abdominal pain in 1.3%, thrombocytopenia in 25%, neutropenia in 13%, leukopenia in 1.3%, hypochromia in 1.3%, elevated serum aspartate aminotransferase in 32.5%, elevated serum alanine aminotransferase in 11.3%, elevated serum bilirubin in 3.8%, decreased serum albumin in 1.3%, elevated serum alkaline phosphatase in 1.3%, elevated gamma-glutamyltranspeptidase in 3.8%, and elevated serum creatinine in 2.5%. Death from myocardial infarction occurred as an incidental event in one case, and no other life-threatening, adverse events were observed. CONCLUSION: Although intra-arterial cisplatin has substantial local and systemic toxicity, high therapeutic efficacy suggests the potential usefulness of this agent in the treatment of advanced HCC. PMID- 18430095 TI - Surgical management of colonic redundancy following esophageal replacement. AB - Colonic redundancy is the most common late complication following esophageal replacement by colonic interposition. Redundancy in the colonic graft leads to mechanical dysfunction of the neo-conduit, causing disabling symptoms that may develop decades after the original surgery. When symptoms caused by food retention in the colonic loop occur, surgical correction may be necessary to improve quality of life and to prevent complications such as aspiration if lifestyle modifications fail. We describe two cases where remedial surgery was performed for redundancy in interposed colonic grafts. Particularly attention is given to preoperative work-up and surgical technique. The literature is reviewed for the etiology, clinical features and management options of this condition. These cases illustrate a successful surgical technique for correcting this complication. PMID- 18430096 TI - Adenocarcinoma developing in de novo Barrett's mucosa in the remnant esophagus after esophagectomy: clinical and molecular assessment. AB - For many patients after subtotal esophagectomy and gastric pull-up, reflux of gastric contents to the esophageal stump is the leading clinical problem. Besides symptoms such as heartburn and regurgitation, de novo formation of columnar mucosa in the esophageal remnant is a well-known and frequent phenomenon. In this context, the remnant supra-anastomotic esophagus serves as an in vivo model for the study of Barrett's carcinogenesis. We present a retrospective case analysis of a patient who developed de novo Barrett's metaplasia followed by de novo invasive carcinoma 28 months after gastric pull-up by assessing clinical and molecular parameters. PMID- 18430097 TI - In memoriam. PMID- 18430098 TI - Impact of hemoglobin level on survival in definitive chemoradiotherapy for T4/M1 lymph node esophageal cancer. AB - We retrospectively investigated the impact of the pre-chemoradiotherapy hemoglobin level (pre-CRT Hb level) for T4 and/or M1 lymph node (LYM) squamous cell carcinoma of the esophagus. Chemotherapy consisted of protracted infusion with 5-fluorouracil (5-FU) at 400 mg/m(2)/day on days 1-5 and 8-12, combined with cisplatin at 40 mg/m(2)/day on days 1 and 8, repeated twice at a 5-week interval. Concurrent radiation therapy was started on day 1 and delivered at 2 Gy/day for five days a week for a total radiation dose of 60 Gy, with a two-week break after a cumulative dose of 30 Gy. Several factors considered to be related with treatment outcome were evaluated by univariate and multivariate analysis. A total of 48 patients with T4/M1 LYM (lymphocyte) esophageal cancer treated with chemoradiotherapy (CRT) between September 2002 and April 2005 were enrolled. The complete response rate to this regimen was 44% and median survival time was 13.6 months, with a median follow-up period of 26.8 months. Median pre-CRT Hb level was 13.5 (10.4-15.3) g/dL. The CR rate in patients with a pre-CRT Hb level of 13 g/dL or less was only 24% but it was 60% in those with a level that was more than 13 g/dL (P=0.01). As for survival, anovarevealed that a pre-CRT Hb of 13 g/dL or less was a significant prognostic factor with a hazard ratio of 0.45 (95% confidence interval [CI]); 0.21-0.97, P=0.04), while on manova, including performance status, tumor size, TNM stage and pre-CRT Hb level, a pre-CRT Hb level of 13 g/dL or less was the only significant prognostic factor, with a hazard ratio of 0.35 (95% CI; 0.13-0.90, P=0.03). In conclusion, the pre-CRT Hb level may be an important determinant of outcome in patients with T4/M1 LYM squamous cell carcinoma of the esophagus. PMID- 18430099 TI - Oxford experience with neoadjuvant chemotherapy and surgical resection for esophageal adenocarcinomas and squamous cell tumors. AB - The Medical Research Council trial for oesophageal cancer (OEO2) trial demonstrated a clear survival benefit from neoadjuvant chemotherapy in resectable esophageal carcinoma. Since February 2000 it has been our practice to offer this chemotherapy regime to patients with T2 and T3 or T1N1 tumors. We analyzed prospectively collected data of patients who received neoadjuvant chemotherapy prior to esophageal resection under the care of a single surgeon. Complications of treatment and overall outcomes were evaluated. A total of 194 patients had cisplatin and 5-fluorouracil prior to esophageal resection. Six patients (5.7%) had progressive disease and were inoperable (discovered in four at surgery). During chemotherapy one patient died and one perforated (operated immediately). Complications including severe neutropenia, coronary artery spasm, renal impairment and pulmonary edema led to the premature cessation of chemotherapy in 12 patients (6.2%). A total of 182 patients with a median age of 63 (range 30 80), 41 squamous and 141 adenocarcinomas underwent surgery. Operations were 91 left thoracoabdominal (50%), 45 radical transhiatal (25%), 40 Ivor-Lewis (22%) and six stage three (3%), and 78.6% had microscopically complete (R0) resections. Median survival was 28 months with 77.3% surviving for 1 year and 57.7% for 2 year. In hospital mortality was 5.5% and anastomotic leak rate 7.7%. A radical surgical approach to the primary tumor in combination with OEO2 neoadjuvant chemotherapy has led to a high R0 resection rate and good survival with acceptable morbidity and mortality. PMID- 18430100 TI - Adjuvant chemotherapy of cisplatin, 5-fluorouracil and leucovorin for complete resectable esophageal cancer: a case-matched cohort study in east China. AB - It is still controversial whether adjuvant chemotherapy of cisplatin, 5 fluorouracil and leucovorin can increase the overall survival of esophageal cancer patients, and which subgroup of patients get most benefits from it. Between 1998 and 2004, 66 esophageal cancer patients with adjuvant chemotherapy and 160 well-matched patients without chemotherapy were included in this study. Nine markers were measured in the protein level to analyze prognostic significance. In the whole group, adjuvant chemotherapy did not improve the survival of esophageal cancer patients. There was also no significant difference for survival in stage I (P=0.59 and P=0.59), stage II (P=0.28 and P=0.28) and stage III patients (P=0.144 and P=0.06) between the observation and the chemotherapy group. Chemotherapy was most effective for the patients who had metastases in cervical and/or celiac lymph nodes (IV subgroup). One and 3-year disease-free survival and overall survival were significantly better than for those who did not receive the chemotherapy(P=0.038, and 0.016, respectively). Bcl 2 expression was a bad prognostic factor, and was more predictive in the adjuvant chemotherapy group than in the no-chemotherapy group. Adjuvant chemotherapy significantly improved the treatment result of stage IV patients compared with the observation group. Bcl-2 could be used to analyze prognosis and guide the adjuvant treatment. PMID- 18430101 TI - Prognostic role of immunosuppressive acidic protein in patients with esophageal cancer. AB - Immunosuppressive acidic protein (IAP) suppresses several immune responses in vivo and in vitro , and high preoperative IAP levels could predict the impairment of the host's immunity. In this study prognostic significance of preoperative IAP levels was investigated in 68 esophageal cancer patients with curative resection and eight with non-curative resection. The curative group had significantly lower levels than the non-curative group (432 +/- 183 mg/mL vs. 739 +/- 235 mg/mL, P < 0.0001). The IAP levels were associated with T-status (P < 0.0001), lymphatic invasion (P < 0.05), and p-stages (P < 0.0001). When 5-year survival rate of patients with curative resection was compared by setting various cutoff values of IAP between high and low IAP groups, several cutoff points (400-580 mg/mL) were revealed to be significantly associated with survival. Setting cutoff value of IAP to 560 mg/mL resulted in a most significant difference of 5-year survival rate of patients between the high and low IAP groups (13.9% and 61.5%, P < 0.0001). These data indicate that pre-operative IAP level is a useful parameter to predict the prognosis of esophageal cancer patients after curative resection. PMID- 18430102 TI - Minimally invasive esophagectomy: a teaching hospital's first year experience. AB - Surgical resection of the esophagus remains the mainstay of treatment for esophageal cancer. However, esophagectomy is associated with significant morbidity and mortality in the postoperative period. We have recently altered our practice pattern to include minimally invasive esophagectomy (MIE) as the approach of choice in the hope of minimizing morbidity associated with this procedure. In this retrospective analysis, we compare outcomes of our first year performing MIE to the previous 3 years of open esophagectomy (OE) at a single teaching hospital. Sixty-five patients underwent esophagectomy between June 2002 and July 2006. Among these, 22 patients underwent MIE between June 2005 and July 2006 and 43 patients underwent OE. The two groups were comparable with regards to age, comorbidities and pathologic stage. The MIE group had less operative blood loss (178 mL vs. 356 mL), decreased respiratory complications requiring mechanical ventila-tion (5% vs. 23%), increased number of lymph nodes procured per procedure (15 vs. 8), and increased number of patients discharged within 10 days (72% vs. 28%) when compared to the OE group. No difference was identified in mortality, complications, or length of stay. PMID- 18430103 TI - The effect of azygos vein preservation on mediastinal lymph node harvesting in thoracic esophagolymphadenectomy. AB - The standard surgical procedure for esophageal cancer is transthoracic esophagectomy with en bloc resection of the azygos vein, thoracic duct and mediastinal lymph nodes. To reduce morbidity of esophago-lymphadenectomy, minimally invasive techniques are increasingly being applied. In (robot-assisted) thoracoscopic esophagolymphadenectomy, the azygos vein is generally left in place, as the scopic ligation of the numerous intercostal veins is technically difficult and time-consuming. This could affect the extent of mediastinal lymph node dissection. Therefore, in this study, the effect of azygos vein preservation during thoracic esophagectomy on mediastinal lymph node harvesting was assessed. In 15 human cadavers, a right-sided thoracotomy was performed, followed by esophagectomy with mediastinal lymph node dissection after ligation of the azygos arch (representing the situation in robot-assisted thoracoscopic esophagolymphadenectomy). Subsequently, the remaining azygos vein with surrounding tissue was resected. The number of lymph nodes in both specimens was determined. A mean of 17.3 (95% Poisson CI 15.3-19.6) lymph nodes was dissected en bloc with the esophagus, and 0.67 (95% Poisson CI 0.32-1.23) around the separately resected azygos vein. The additional azygos vein resection did not add to the number of lymph nodes dissected in 60% (9/15) of cadavers. In conclusion, the extent of mediastinal lymph node dissection was not substantially affected by leaving the azygos vein in situ . Time-sparing azygos vein preservation in (robot assisted) thoracoscopic esophagolymphadenectomy may therefore be considered justified. PMID- 18430104 TI - Gene expression profiling of the response of esophageal carcinoma cells to cisplatin. AB - Cisplatin is the most common chemotherapeutic agent used in esophageal cancer. However, sensitivity to cisplatin varies greatly between patients. It is important to identify the gene(s) that are related to the sensitivity to cisplatin in esophageal cancer patients. The IC50 for cisplatin was measured for 15 esophageal cancer cell lines (TE1-5, TE8-15, KYSE140, and KYSE150). RNA was extracted from each of these cell lines and a normal esophageal epithelial cell line, namely, Het1A, and gene expression profiles were analyzed using an oligonucleotide microarray consisting of 34 594 genes. TE4 was highly resistant and TE12, 14, and 15 were sensitive to cisplatin. Thirty-seven genes were differentially expressed in the cisplatin-resistant esophageal cancer cell line. Our investigation provides a list of candidate genes that may be associated with resistance to cisplatin in esophageal cancer cells, which may serve as a basis for additional functional studies. PMID- 18430105 TI - The prognostic impact of occult lymph node metastasis in cancer of the esophagus or esophago-gastric junction: systematic review and meta-analysis. AB - Attempts to define the clinical significance of occult lymph node metastasis have yielded mixed results. We set out to quantify the influence on disease-free survival of occult lymph node metastasis in cases of esophageal or gastro esophageal cancer previously staged as lymph node-negative by conventional H&E staining. We performed a systematic review and meta-analysis of observational studies published between 1966 and 2006 (identified through Medline and Embase). Twelve suitable cohort studies were identified. These studies suggest there is a significant (P < 0.001) association between occult lymph node metastasis and prognosis in cancer of the esophagus or esophago-gastric junction (pooled hazard ratio 3.16 with 95% confidence intervals of 2.25-4.42). We did not demonstrate study quality, number of nodes examined or number of lymph node sections examined to be significant sources of intertrial heterogeneity. Data from observational studies suggest that occult lymph node metastasis is an important prognostic factor in cancer of the esophagus or gastro-esophagus. Meta-analysis using individual patient data can now be justified. PMID- 18430107 TI - Endoscopic findings in a cohort of newly diagnosed gastroesophageal reflux disease patients registered in a UK primary care database. AB - Gastroesophageal reflux disease (GERD) may be accompanied by erosive complications that are diagnosed by endoscopy. This study aimed to describe the characteristics of patients newly diagnosed with GERD who are referred for endoscopy, and the factors associated with esophageal endoscopic findings. This study included patients aged 2-79 years with a first recorded diagnosis of GERD in 1996, as identified in a previous cohort study in the UK General Practice Research Database. The rate and results of endoscopy were recorded. Unconditional logistic regression analysis was used to estimate the odds ratios and 95% confidence intervals for the relationship between a range of factors and endoscopy and its findings. Of the 7159 patients with a new GERD diagnosis, 805 (11%) underwent endoscopy close to the time of first consultation for GERD. Endoscopic findings indicative of esophageal damage were recorded in 73% of these patients. Esophageal endoscopic findings were significantly more likely in males, older patients, and individuals with a history of peptic ulcer disease or gastrointestinal bleeding. Use of acid-suppressive drugs, particularly proton pump inhibitors, was inversely associated with erosive endoscopic findings. Patients with erosive endoscopic findings were more likely to start a new course of treatment with a proton pump inhibitor. In conclusion, relatively few patients are referred for endoscopy close to the first consultation for GERD, and the majority of these individuals have esophageal findings. Male gender, increasing age and a history of bleeding were risk factors for esophageal complications. PMID- 18430106 TI - Impact of endoscopic ultrasonography and physician specialty on the management of patients with esophagus cancer. AB - While endoscopic ultrasonography (EUS) and EUS-guided fine-needle aspiration (EUS FNA) are the most accurate techniques for locoregional staging of esophageal cancer, little evidence exists that these innovations impact on clinical care. The objective on this study was to determine the frequency with which EUS and EUS FNA alter the management of patients with localized esophageal cancer, and assess practice variation among specialists at a tertiary care center. Three gastroenterologists, three medical oncologists, three radiation oncologists and four thoracic surgeons were asked to independently report their management recommendations as the anonymized staging information of 50 prospectively enrolled patients from another study were sequentially disclosed on-line. Compared to initial management recommendations, that were based upon history, physical examination, upper endoscopy and CT scan results, EUS prompted a change in management 24% (95% CI: 12-36%) of the time; usually to a more resource intensive approach (71%), for example from recommending palliation to recommending neoadjuvant chemoradiation therapy. EUS-FNA plus cytology results altered management an additional 8% (95% CI: 6-15%) of the time. Agreement between specialists ranged from fair (intraclass correlation [ICC=0.32) to substantial (ICC=0.65); improving with additional information. Among specialists, agreement was greatest for patients with stage I disease. EUS and EUS-FNA changed patient management the most for patients with stages IIA, IIB or III disease. EUS, with or without FNA, significantly impacts the management of patients with localized esophageal cancer. With respect to the optimal treatment for each patient, agreement among physicians incrementally increases with endoscopic ultrasound results. Specialty training appears to influence therapeutic decision making behavior. PMID- 18430108 TI - Persistent dysphagia after laparoscopic fundoplication for gastro-esophageal reflux disease. AB - Persistent postoperative dysphagia is a potentially severe complication of fundoplication for gastroesophageal reflux disease (GERD). The aim of this retrospective study was to analyze our experience of laparoscopic fundoplication for GERD in 276 consecutive patients, to determine the frequency of postoperative dysphagia and assess treatments and outcomes. There was no relation between preoperative dysphagia, present in 24 patients (8.7%), and postoperative DeMeester grade 2 or 3 dysphagia, present in 25 patients (9.1%). Ten (3.6%) patients had clinically significant postoperative dysphagia, eight (2.9%) underwent esophageal dilation, with symptom improvement in five. Four (1.4%) of our patients (two with failed dilation) and 11 patients receiving antireflux surgery elsewhere, underwent re-operation for persistent dysphagia 12 months (median) after the first operation. DeMeester grade 0 or 1 dysphagia was obtained in 10/13 evaluable patients. Our experience is fully consistent with that of the recent literature. Redo surgery is necessary in only a small fraction of operated patients with GERD with good probability of resolving the dysphagia. Best outcomes are obtained when an anatomical cause of the dysphagia is documented preoperatively. PMID- 18430109 TI - A new endoscopic technique for suspension of esophageal prosthesis for refractory caustic esophageal strictures. AB - There is no clear consensus concerning the best endoscopic treatment of benign refractory esophageal strictures due to caustic ingestion. Different procedures are currently used: frequent multiple dilations, retrievable self-expanding stent, nasogastric intubation and surgery. We describe a new technique to fix a suspended esophageal silicone prosthesis to the neck in benign esophageal strictures; this permits us to avoid the frequent risk of migration of the expandable metallic or plastic stents. Under general anesthesia a rigid esophagoscope was placed in the patient's hypopharynx. Using transillumination from the optical device, the patient's neck was pierced with a needle. A n.0 monofilament surgical wire was pushed into the needle, grasped by a standard foreign body forceps through the esophagoscope and pulled out of the mouth (as in percutaneous endoscopic gastrostomy procedure). After tying the proximal end of the silicone prosthesis with the wire, it was placed through the strictures under endoscopic view. This procedure was successfully utilized in four patients suffering from benign refractory esophageal strictures due to caustic ingestion. The prosthesis and its suspension from the neck were well-tolerated until removal (mean duration 4 months). A postoperative transitory myositis was diagnosed in only one patient. One of the most frequent complications of esophageal prostheses in refractory esophageal strictures due to caustic ingestion is distal migration. Different solutions were proposed. For example the suspension of a wire coming from the nose and then fixed behind the ear. This solution is not considered optimal because of patient complaints and moreover the aesthetic aspect is compromised. The procedure we utilized in four patients utilized the setting of a silicone tube hanging from the neck in a way similar to that of endoscopic pharyngostomy. This solution is a valid alternative both for quality of life and for functional results. PMID- 18430110 TI - Risk factors for surgical site infection after surgery for esophageal perforation. AB - Esophageal perforations carry a high potential for morbidity and mortality. The prognosis depends on rapid and precise diagnosis and management. Surgical site infections (SSIs) are very common following the surgical treatment of esophageal lesions. We aimed identify significant risk factors for SSI after surgery for esophageal perforation via an historical cohort study including patients who underwent surgical management of esophageal perforation. The predictive variables were analyzed by bivariate analysis and multiple logistic regression. Eighty-one patients were studied during a 10-year period ending in 2004. The mean age was 42.6 years. In 44% of the patients the time interval between the perforation and surgery was up to 6 h and in 30% it was > 24 h. Associated lesions occurred in other cavities; 17% in the chest, 5% in the abdomen, 5% in the extremities, 4% in the spinal column and bone marrow and 2% in the face. There were grade I lesions in eight cases (10%), grade II in 64 cases (79%) and grade III in nine cases (11%). The mean time of surgery procedure was 117.2 min. The mean SSI was 7.99. SSIs occurred in 33 patients (41%). The risk factors for SSI following surgical management of esophageal perforation were: age > or = 50 years, time delay to treatment > 24 h, associated lesion in another cavity and Injury Severity Score > or = 15. PMID- 18430111 TI - Vascular anatomy of the stomach related to gastric tube construction. AB - In view of constructing a gastric tube after esophagus resection, the vascular anatomy of the greater curvature of the stomach, especially the connection between the left and right gastro-epiploic arteries, was investigated. The vascular anatomy was studied in 20 embalmed human specimens. After dissection a gastric tube of 4 cm wide was constructed, using the greater gastric curvature. Various lengths of the arterial arcades were measured. In 70% an anastomosis between the right and left gastro-epiploic arteries was present. With the construction of an isoperistaltic gastric tube, in which the left gastro-epiploic artery is left in situ (ligating it at the splenic hilus), there is an 18.7% increase of length of arterial arcade along the gastric tube. Leaving the left gastro-epiploic artery in situ increases the feeding arterial arcaded-length along the gastric tube with 5.0 cm (19%). PMID- 18430112 TI - A new operative technique for the resection of gastric tube cancer by means of lifting the anterior chest wall and videoscope-assisted surgery. AB - The prolonged survival of patients receiving surgery for esophageal cancer has led to an increased incidence of adenocarcinoma arising in the gastric tube used for reconstruction (gastric tube cancer). In patients with advanced gastric tube cancer, resection of the gastric tube should be considered, but currently available procedures are very invasive. In patients undergoing curative surgery for gastric tube cancer that has developed after reconstruction through the retrosternal route, the gastric tube is usually resected through a median sternotomy, followed by reconstruction with the colon. However, postoperative complications often occur and treatment outcomes remain poor. We developed a new surgical technique for gastric tube resection without performing a sternotomy in patients with gastric tube cancer who had previously undergone reconstruction through the retrosternal route. Our technique was used to treat two patients. Two Kirschner wires were passed subcutaneously through the anterior chest; the chest was lifted to extend the retrosternal space and secure an adequate surgical field. The stomach was separated from the surrounding tissue under videoscopic guidance. Total resection of the gastric tube was done. The retrosternal space was used to lift the jejunum. Roux-en-Y reconstruction was performed. Neither patient had suture line failure or surgical site infection. Their recovery was uneventful. Our surgical technique has several potential advantages including (i) reduced surgical stress; (ii) the ability to use the retrosternal space for reconstruction after gastric tube resection; and (iii) a reduced risk of serious infections such as osteomyelitis in patients with suture line failure. Our findings require confirmation by additional studies. PMID- 18430114 TI - Carbonic anhydrase IX: role in diagnosis prognosis and cancer therapy. Introduction. PMID- 18430115 TI - Carbonic anhydrase IX: historical and future perspectives. PMID- 18430116 TI - Molecular mechanisms of carbonic anhydrase IX-mediated pH regulation under hypoxia. PMID- 18430117 TI - Significance of pH regulation and carbonic anhydrases in tumour progression and implications for diagnostic and therapeutic approaches. PMID- 18430118 TI - Cancer-associated, hypoxia-inducible carbonic anhydrase IX facilitates CO2 diffusion. PMID- 18430119 TI - Carbonic anhydrase IX and renal cell carcinoma: prognosis, response to systemic therapy, and future vaccine strategies. PMID- 18430120 TI - Carbonic anhydrase IX as a predictive biomarker of response to kidney cancer therapy. PMID- 18430121 TI - The use of positron-emission tomography in the diagnosis of tumour phenotype. PMID- 18430122 TI - Development of small molecule carbonic anhydrase IX inhibitors. PMID- 18430123 TI - Immunohistochemical expression of carbonic anhydrase IX assessed over time and during treatment in renal cell carcinoma. PMID- 18430124 TI - The role of carbonic anhydrase IX as a molecular marker for transitional cell carcinoma of the bladder. PMID- 18430125 TI - A perspective on liposomal amphotericin B (AmBisome). PMID- 18430126 TI - Changing epidemiology of systemic fungal infections. AB - Species of Candida and Aspergillus remain the most common causes of invasive fungal infections, but other yeasts and filamentous fungi are emerging as significant pathogens. Opportunistic yeast-like fungi and moulds such as Zygomycetes, Fusarium spp. and Scedosporium spp. are increasingly being recognised in patient groups such as those with leukaemia and in bone marrow transplant recipients. Recognition of these epidemiological changes is critical to patient care. The key elements in selecting an appropriate antifungal agent are the type of patient (solid-organ or stem-cell transplant), severity of immunosuppression, history of prolonged exposure to antifungal drugs, and knowledge of the genera and species of the infecting pathogen and its typical susceptibility pattern. PMID- 18430127 TI - Amphotericin B lipid preparations: what are the differences? AB - To reduce the in-vivo toxicity of the broad-spectrum antifungal drug amphotericin B, various lipid formulations of amphotericin B, ranging from lipid complexes to small unilamellar liposomes, have been developed and subsequently commercialized. These structurally diverse formulations differ in their serum pharmacokinetics as well as their tissue localisation, tissue retention and toxicity. These differences can affect the choice of formulation for a given infection, the time of initiation of treatment, and the dosing regimen. Although preclinical studies have shown similarities in the in-vitro and in-vivo antifungal activity of the formulations with comparable dosing, their acute and chronic toxicity profiles are not the same, and this has a significant impact on their therapeutic indices, especially in high-risk, immunosuppressed patients. With the recent introduction of new antifungal drugs to treat the increasing numbers of infected patients, the amphotericin B lipid formulations are now being studied to evaluate their potential in combination drug regimens. With proven efficacy demonstrated during the past decade, it is expected that amphotericin B lipid formulations will remain an important part of antifungal drug therapy. PMID- 18430128 TI - Contribution of new diagnostic approaches to antifungal treatment plans in high risk haematology patients. AB - In high-risk patient cohorts, such as patients after solid-organ or allogeneic stem-cell transplantation, or patients with acute leukaemia, early diagnosis of invasive fungal infections (IFIs) is essential, as delayed or missing diagnosis of IFI results in increasing rates of mortality. However, diagnosis of most IFIs, especially of invasive aspergillosis, is difficult because classic tests have low sensitivity and specificity, and radiology often provides non-specific and transient results. The limited sensitivity and specificity of conventional assays for the detection of IFI and the growing number of immunocompromised patients who are at risk for opportunistic fungal infections have led to the development of new assays. These methods include antigen detection systems, such as ELISAs, and different molecular methods (PCR assays). Serological tests, such as the detection of the carbohydrate galactomannan, are standardised and commercially available. However, they still need to be evaluated in large patient cohorts, especially children. The benefit of antibody detection remains unclear if patients are under immune suppression or are heavily colonised but not infected. A range of different PCR assays (conventional, nested, real-time) have been developed, targeting different gene regions (cytochrome P450, heat-shock proteins, 18S, 5.8S, 28S, internal transcribed spacer), including a variety of amplicon detection methods, such as gel electrophoresis, hybridisation with specific probes, ELISA and restriction fragment length polymorphism. These molecular assays provide high potential in terms of sensitivity and specificity, but vary widely in their feasibility and up to now have not been standardised. Taken together, new non-culture-based diagnostic assays are appropriate as simple and rapid screening tests with high sensitivities and quick turnaround times. Thus, they might help to reduce empirical antifungal therapy and might be valuable tools to allow early initiation and monitoring of pre-emptive antifungal therapy. In this review, we assess the performance of a variety of non-culture based tests for the detection of IFI in high-risk haematological patients, with emphasis on the impact of the assays on different management strategies. PMID- 18430129 TI - New options for treatment of candidaemia in critically ill patients. AB - Bloodstream infections caused by Candida spp. are increasingly recognised in critically ill adult and paediatric individuals, with significant associated morbidity and mortality. Candida albicans is the single most common fungal species to cause nosocomial infections. However, non-C. albicans spp., including Candida glabrata and Candida krusei, which are less susceptible to fluconazole, have become more common. Until the 1980s, the therapeutic possibilities for invasive candidosis were limited to amphotericin B, but with the advent of new antifungal agents, such as azoles and echinocandins, less toxic therapeutic options have become available and there are now possibilities for prevention and optimised therapy for documented Candida infections. In this review, the currently available options for the treatment of candidaemia and invasive candidosis are discussed with regard to the role of liposomal amphotericin B in comparison with the echinocandins and azoles. PMID- 18430130 TI - New dosing strategies for liposomal amphotericin B in high-risk patients. AB - In recent years, the focus of attention in our understanding of pharmacokinetic antifungal drug efficacy has expanded from the vascular to the tissue compartment, since for moulds in particular, this is the primary point of encounter between the invading fungus and the host. Liposomal amphotericin B (LAB) accumulates in the reticulo-endothelial system and other tissues for several weeks after systemic administration at concentrations exceeding the MICs for many pathogenic fungi. Animal models demonstrate that such tissue depots provide effective prophylaxis and even therapeutic opportunities when LAB is given in high intermittent doses. Efficacy has been shown for even a single high dose of LAB. Human studies have also confirmed retention of amphotericin B in tissues well beyond the last administered dose. Clinical evidence has begun to accrue that suggests prophylactic efficacy in high-risk patients with haematological malignancies who have received intermittent LAB. In an exploratory study of patients with persistent and protracted neutropenic fever, one dose of 10 mg/kg, followed by two doses of 5 mg/kg given on days 1, 3 and 6, respectively, appeared to be as effective as the standard regimen of 3 mg/kg/day given for a longer period. Serum kinetics suggest a large-volume deep tissue compartment for LAB. The drug also appears to accumulate in the tissue, as reflected by bone marrow concentrations. These early observations suggest the potential for intermittent high dosing of LAB for prophylaxis and management of invasive fungal infections, thus providing an alternative option to more frequent and expensive administration of LAB, and daily administration of azoles or candins. This might offer the benefits of lower treatment costs, improved patient compliance and reduced toxicity. Further clinical studies are required to confirm the feasibility of such an approach. PMID- 18430131 TI - Combination antifungal therapy: a critical review of the evidence. AB - Invasive fungal infections have extremely high rates of morbidity and mortality, particularly in immunocompromised hosts. Combination antifungal therapy is conceptually attractive as a life-saving measure. However, in-vitro and in-vivo evidence is often conflicting and clinical trials in this area are limited. Most clinical studies show similar outcomes for combination antifungal therapy when compared to monotherapy, although secondary endpoints and sub-analyses often show advantages for the combinations in endpoints such as culture sterilisation. The logistics of large clinical trials of combination therapy are highly complex. Combination of antifungals with immune modulators is an exciting new research area. Until more data are available, clinicians should approach combination antifungal therapy with caution. PMID- 18430132 TI - Liposomal amphotericin B: what is its role in 2008? AB - Although our antifungal armamentarium has been enlarged recently with new azoles (voriconazole and posaconazole) and echinocandins (caspofungin, micafungin, anidulafungin), the polyenes still have an important role in antifungal strategies because of their extended antifungal spectrum and rarity of mycological resistance. The use of conventional amphotericin B deoxycholate is limited by substantial toxicity that is either infusion-related or associated with renal failure. Its lipid derivatives, particularly liposomal amphotericin B (LAmB), are less nephrotoxic while maintaining a broad antifungal spectrum. LAmB is active against most Candida spp., including Candida glabrata and Candida parapsilosis, and against more resistant, emerging yeasts species such as Rhodotorula spp., Geotrichum spp. and Trichosporon spp.. LAmB is also active against Cryptococcus spp. and all dimorphic fungi such as Histoplasma, Blastomyces, Coccidioidomyces, and Paracoccidiodomyces. The antifungal spectrum of LAmB is particularly interesting with regard to filamentous fungi, with marked activity against Aspergillus spp. and agents of zygomycosis. The latter might emerge during long-term treatment with voriconazole or an echinocandin, as these organisms are resistant to these drugs. We review here the role of LAmB in the current antifungal management strategy, which is based on results obtained in prospective trials. LAmB can be retained as first-line treatment for human immunodeficiency virus (HIV)-positive patients with disseminated histoplasmosis and cryptococcosis, even in the setting of renal impairment or concomitant administration of potentially nephrotoxic drugs. In addition, there is sufficient evidence that the drug should be a major consideration for the empirical treatment of persistent febrile neutropenia or as an alternative to for patients with invasive aspergillosis, for those at risk of renal impairment, major drug drug interaction or liver insufficiency, particularly in the situation of an established azole intolerance. The primary licensed indication for LAmB is empirical treatment. When zygomycosis is suspected or has been documented, high doses of LAmB should be prescribed. Finally, LAmB may also be considered as a therapeutic option for the management of candidaemia and remains a cornerstone for the treatment of some visceral localisations during systemic candidosis. PMID- 18430133 TI - A novel regulatory protein governing biofilm formation in Bacillus subtilis. AB - Production of an extracellular matrix is a hallmark of biofilm formation. In the spore-forming bacterium Bacillus subtilis, the matrix consists of an exopolysaccharide, which is specified by the epsA-O operon, and a secreted protein TasA, which is encoded by the yqxM-sipW-tasA operon. Past and present evidence establish that the epsA-O and yqxM-sipW-tasA operons are controlled by the repressor proteins SinR and AbrB. Here, we report the identification of a novel regulatory protein Slr that promotes transcription of the yqxM-sipW-tasA operon but is not needed for expression of the epsA-O operon. We further show that the gene for Slr is itself under the negative control of SinR and AbrB. These findings reveal that matrix production is governed by an intricate network involving the interplay of negatively and positively acting regulatory proteins. PMID- 18430134 TI - The receiver domain of FrzE, a CheA-CheY fusion protein, regulates the CheA histidine kinase activity and downstream signalling to the A- and S-motility systems of Myxococcus xanthus. AB - The Frz chemosensory system is a two-component signal transduction pathway that controls cell reversals and directional movements for the two motility systems in Myxococcus xanthus. To trigger cell reversals, FrzE, a hybrid CheA-CheY fusion protein, autophosphorylates the kinase domain at His-49, and phosphoryl groups are transferred to aspartate residues (Asp-52 and Asp-220) in the two receiver domains of FrzZ, a dual CheY-like protein that serves as the pathway output. The role of the receiver domain of FrzE was unknown. In this paper, we characterize the FrzE protein in vitro and show that the receiver domain of FrzE negatively regulates the autophosphorylation activity of the kinase domain of FrzE. Unexpectedly, it does not appear to play a direct role in phospho-relay as in most other histidine kinase receiver domain hybrid systems. The regulatory role of the FrzE receiver domain suggests that it may interact with or be phosphorylated by an unknown protein. We also show the dynamics of motility system-specific marker proteins in FrzE mutants as cells move forward and reverse. Our studies indicate that the two motility systems are functionally co ordinated and that any system-specific branching of the pathway most likely occurs downstream of FrzE. PMID- 18430136 TI - Fold and function of polypeptide transport-associated domains responsible for delivering unfolded proteins to membranes. AB - Membranes of Gram-negative bacteria, mitochondria and chloroplasts receive and fold beta-barrel transmembrane proteins through the action of polypeptide transport-associated (POTRA) domains. In Escherichia coli, folding substrates are inserted into the outer membrane by the essential protein YaeT, a prototypic Omp85 protein. Here, the articulation between tandem POTRA domains in solution is defined by nuclear magnetic resonance (NMR) spectroscopy, indicating an unprecedented juxtaposition. The novel solution orientations of all five POTRA domains are revealed by small-angle X-ray scattering of the entire 46 kDa periplasmic region. NMR titration studies show that strands from YaeT's canonical folding substrate, PhoE, bind non-specifically along alternating sides of its mixed beta sheets, thus providing an ideal platform for helping to fold nascent outer-membrane proteins. Together, this provides the first structural model of how multiple POTRA domains recruit substrates from the periplasmic solution into the outer membrane. PMID- 18430135 TI - The global, ppGpp-mediated stringent response to amino acid starvation in Escherichia coli. AB - The stringent response to amino acid starvation, whereby stable RNA synthesis is curtailed in favour of transcription of amino acid biosynthetic genes, is controlled by the alarmone ppGpp. To elucidate the extent of gene expression effected by ppGpp, we designed an experimental system based on starvation for isoleucine, which could be applied to both wild-type Escherichia coli and the multiauxotrophic relA spoT mutant (ppGpp(0)). We used microarrays to profile the response to amino acid starvation in both strains. The wild-type response included induction of the general stress response, downregulation of genes involved in production of macromolecular structures and comprehensive restructuring of metabolic gene expression, but not induction of amino acid biosynthesis genes en masse. This restructuring of metabolism was confirmed using kinetic Biolog assays. These responses were profoundly altered in the ppGpp(0) strain. Furthermore, upon isoleucine starvation, the ppGpp(0) strain exhibited a larger cell size and continued growth, ultimately producing 50% more biomass than the wild-type, despite producing a similar amount of protein. This mutant phenotype correlated with aberrant gene expression in diverse processes, including DNA replication, cell division, and fatty acid and membrane biosynthesis. We present a model that expands and functionally integrates the ppGpp-mediated stringent response to include control of virtually all macromolecular synthesis and intermediary metabolism. PMID- 18430137 TI - Mini-III, a fourth class of RNase III catalyses maturation of the Bacillus subtilis 23S ribosomal RNA. AB - Ribonuclease III (RNase III) type of enzymes are double-stranded RNA (dsRNA) specific endoribonucleases that have important roles in RNA maturation and mRNA decay. They are involved in processing precursors of ribosomal RNA (rRNA) in bacteria as well as precursors of short interfering RNAs (siRNAs) and microRNAs (miRNAs) in eukaryotes. RNase III proteins have been grouped in three major classes according to their domain organization. In this issue of Molecular Microbiology, Redko et al. identified a novel class of bacterial RNase III, named Mini-III, consisting only of the RNase III catalytic domain and functioning in the maturation of the 23S rRNA in Bacillus subtilis. Its absence from proteobacteria reveals that this step is mechanistically different from the corresponding step in Escherichia coli. The fact that Mini-III orthologues are present in unicellular photosynthetic eukaryotes and in plants opens new opportunities for functional studies of this type of RNases. PMID- 18430138 TI - The type III secretion system tip complex and translocon. AB - The type III secretion machinery of Gram-negative bacteria, also known as the injectisome or needle complex, is composed of a basal body spanning both bacterial membranes and the periplasm, and an external needle protruding from the bacterial surface. A set of three proteins, two hydrophobic and one hydrophilic, are required to allow translocation of proteins from the bacterium to the host cell cytoplasm. These proteins are involved in the formation of a translocation pore, the translocon, in the host cell membrane. Exciting progress has recently been made on the interaction between the translocators and the injectisome needle and the assembly of the translocon in the host cell membrane. As expected, the two hydrophobic translocators insert into the target cell membrane. Unexpectedly, the third, hydrophilic translocator, forms a complex on the distal end of the injectisome needle, the tip complex, and serves as an assembly platform for the two hydrophobic translocators. PMID- 18430139 TI - Lipid spirals in Bacillus subtilis and their role in cell division. AB - The fluid mosaic model of membrane structure has been revised in recent years as it has become evident that domains of different lipid composition are present in eukaryotic and prokaryotic cells. Using membrane binding fluorescent dyes, we demonstrate the presence of lipid spirals extending along the long axis of cells of the rod-shaped bacterium Bacillus subtilis. These spiral structures are absent from cells in which the synthesis of phosphatidylglycerol is disrupted, suggesting an enrichment in anionic phospholipids. Green fluorescent protein fusions of the cell division protein MinD also form spiral structures and these were shown by fluorescence resonance energy transfer to be coincident with the lipid spirals. These data indicate a higher level of membrane lipid organization than previously observed and a primary role for lipid spirals in determining the site of cell division in bacterial cells. PMID- 18430141 TI - A Csr-type regulatory system, including small non-coding RNAs, regulates the global virulence regulator RovA of Yersinia pseudotuberculosis through RovM. AB - The MarR-type regulator RovA controls expression of virulence genes of Yersinia pseudotuberculosis in response to environmental signals. Using a genetic strategy to discover components that influence rovA expression, we identified new regulatory factors with homology to components of the carbon storage regulator system (Csr). We showed that overexpression of a CsrB- or a CsrC-type RNA activates rovA, whereas a CsrA-like protein represses RovA synthesis. We further demonstrate that influence of the Csr system on rovA is indirect and occurs through control of the LysR regulator RovM, which inhibits rovA transcription. The CsrA protein had also a major influence on the motility of Yersinia, which was independent of RovM. The CsrB and CsrC RNAs are differentially expressed in Yersinia. CsrC is highly induced in complex but not in minimal media, indicating that medium-dependent rovM expression is mediated through CsrC. CsrB synthesis is generally very low. However, overexpression of the response regulator UvrY was found to activate CsrB production, which in turn represses CsrC synthesis independent of the growth medium. In summary, the post-transcriptional Csr-type components were shown to be key regulators in the co-ordinated environmental control of physiological processes and virulence factors, which are crucial for the initiation of Yersinia infections. PMID- 18430142 TI - Identification and characterization of KpsS, a novel polysaccharide sulphotransferase in Mesorhizobium loti. AB - Plants enter into symbiotic relationships with bacteria that allow survival in nutrient-limiting environments. The bacterium Mesorhizobium loti enters into a symbiosis with the legume host, Lotus japonicus, which results in the formation of novel plant structures called root nodules. The bacteria colonize the nodules, and are internalized into the cytoplasm of the plant cells, where they reduce molecular dinitrogen for the plant. Symbiosis between M. loti and L. japonicus requires bacterial synthesis of secreted and cell-surface polysaccharides. We previously reported the identification of an unusual sulphate-modified form of capsular polysaccharide (KPS) in M. loti. To better understand the physiological function of sulphated KPS, we isolated the sulphotransferase responsible for KPS sulphation from M. loti extracts, determined its amino acid sequence and identified the corresponding M. loti open reading frame, mll7563 (which we have named kpsS). We demonstrated that partially purified KpsS functions as a fucosyl sulphotransferase in vitro. Furthermore, mutants deficient for this gene exhibit a lack of KPS sulphation and a decreased rate of nodule formation on L. japonicus. Interestingly, the kpsS gene product shares no significant amino acid similarity with previously identified sulphotransferases, but exhibited sequence identity to open reading frames of unknown function in diverse bacteria that interact with eukaryotes. PMID- 18430140 TI - Trypanosoma brucei BRCA2 acts in antigenic variation and has undergone a recent expansion in BRC repeat number that is important during homologous recombination. AB - Antigenic variation in Trypanosoma brucei has selected for the evolution of a massive archive of silent Variant Surface Glycoprotein (VSG) genes, which are activated by recombination into specialized expression sites. Such VSG switching can occur at rates substantially higher than background mutation and is dependent on homologous recombination, a core DNA repair reaction. A key regulator of homologous recombination is BRCA2, a protein that binds RAD51, the enzyme responsible for DNA strand exchange. Here, we show that T. brucei BRCA2 has undergone a recent, striking expansion in the number of BRC repeats, a sequence element that mediates interaction with RAD51. T. brucei BRCA2 mutants are shown to be significantly impaired in antigenic variation and display genome instability. By generating BRCA2 variants with reduced BRC repeat numbers, we show that the BRC expansion is crucial in determining the efficiency of T. brucei homologous recombination and RAD51 localization. Remarkably, however, this appears not to be a major determinant of the activation of at least some VSG genes. PMID- 18430143 TI - Speciation in the highlands of Mexico: genetic and phenotypic divergence in the Mexican jay (Aphelocoma ultramarina). AB - The pine-oak woodlands of the Mexican highlands harbour significant biological diversity, yet little is known about the evolutionary history of organisms inhabiting this region. We assessed genetic and phenotypic differentiation in 482 individuals representing 27 populations of the Mexican jay (Aphelocoma ultramarina) - a widespread bird species of the Mexican highlands - to test whether populations in the central and northern Mexican sierras display discrete breaks between groups, which would be consistent with a role for the different mountain chains in divergence and speciation. We found abrupt breaks in mitochondrial DNA (mtDNA; ND2 and control region) delineating four major genetic groups found in the Sierra Madre Occidental, Sierra Madre Oriental, southern Central Plateau (Bajio), and Transvolcanic Belt. These mtDNA groups were largely corroborated by data from nuclear microsatellites and phenotypic data, except that clades from the Central Plateau and Sierra Madre Oriental showed clinal change in these data sets. Uncertainty about the mutation rate for our mitochondrial markers warrants considerable caution with regard to estimating divergence times, but the major genetic groups appear to have split before the most extreme period of glacial cycling that marked the last 0.7 million years and after Mexico's period of major mountain formation. The fact that some genetic breaks do not coincide with well-known geographic barriers suggests a role for ecology in divergence and speciation, and we discuss implications for taxonomy and conservation. PMID- 18430145 TI - Landscape genetics and hierarchical genetic structure in Atlantic salmon: the interaction of gene flow and local adaptation. AB - Disentangling evolutionary forces that may interact to determine the patterns of genetic differentiation within and among wild populations is a major challenge in evolutionary biology. The objective of this study was to assess the genetic structure and the potential influence of several ecological variables on the extent of genetic differentiation at multiple spatial scales in a widely distributed species, the Atlantic salmon, Salmo salar. A total of 2775 anadromous fish were sampled from 51 rivers along the North American Atlantic coast and were genotyped using 13 microsatellites. A Bayesian analysis clustered these populations into seven genetically and geographically distinct groups, characterized by different environmental and ecological factors, mainly temperature. These groups were also characterized by different extent of genetic differentiation among populations. Dispersal was relatively high and of the same magnitude within compared to among regional groups, which contrasted with the maintenance of a regional genetic structure. However, genetic differentiation was lower among populations exchanging similar rates of local as opposed to inter regional migrants, over the same geographical scale. This raised the hypothesis that gene flow could be constrained by local adaptation at the regional scale. Both coastal distance and temperature regime were found to influence the observed genetic structure according to landscape genetic analyses. The influence of other factors such as latitude, river length and altitude, migration tactic, and stocking was not significant at any spatial scale. Overall, these results suggested that the interaction between gene flow and thermal regime adaptation mainly explained the hierarchical genetic structure observed among Atlantic salmon populations. PMID- 18430144 TI - Differential gene expression in incipient species of Anopheles gambiae. AB - A speciation process is ongoing in the primary vector of malaria in Africa, Anopheles gambiae. Assortatively mating incipient species known as the M and S forms differentially exploit larval breeding sites associated with different ecological settings. However, some ongoing gene flow between M and S limits significant genomic differentiation mainly to small centromere-proximal regions on chromosomes X and 2L, termed 'speciation islands' with the expectation that they contain the genes responsible for reproductive isolation. As the speciation islands exhibit reduced recombination and low polymorphism, more detailed genetic analysis using fine-scale mapping is impractical. We measured global gene expression differences between M and S using oligonucleotide microarrays, with the goal of identifying candidate genes that could be involved in this ongoing speciation process. Gene expression profiles were examined in two independent colonies of both forms at each of three developmental periods of interest: fourth instar larvae, virgin females, and gravid females. Patterns were validated on a subset of genes using quantitative real-time reverse transcription polymerase chain reaction of RNA samples from laboratory colonies and wild mosquitoes collected from Cameroon and Burkina Faso. Considered across all three developmental periods, differentially expressed genes represented approximately 1 2% of all expressed genes. Although disproportionately represented in the X speciation island, the vast majority of genes were located outside any speciation island. Compared to samples from the other developmental periods, virgin females were characterized by more than twice as many differentially expressed genes, most notably those implicated in olfaction and potentially, mate recognition. PMID- 18430146 TI - High frequency of multiple paternity in broods of a socially monogamous cichlid fish with biparental nest defence. AB - In several animal taxa, genetic analyses have demonstrated that social monogamy and biparental brood care do not preclude polygamous reproduction. Few studies have been conducted in fish, but in fish species without alternative reproductive phenotypes, social monogamy was largely congruent with genetic parentage. In contrast to these findings, we report an exceptionally high level of multiple paternity in a socially monogamous cichlid fish with biparental nest defence (Variabilichromis moorii), inferred from microsatellite and mitochondrial data of 10 broods. Whereas all offspring in a nest shared a common mother, each brood was sired by 2 to > 10 males. None of the inferred sires was assigned a large proportion of the brood. Paternity was estimated as the minimum number of sires required to explain multilocus offspring genotypes, and as the maximum-likelihood number of sires given population allele frequencies. Analysis of simulated brood genotypes suggested that, although these two methods tend to under- and overestimate, respectively, the true number of sires, primary sires with many offspring in a brood would have been detected. Hence, the genetic data indicate that the nest tending males suffer substantial cuckoldry and provide alloparental care for a large number of unrelated fry. We have no data on the social status of the cuckolding males, but due to synchronous spawning of pairs and commitment to brood care of paired males, it is possible that most of the parasitic spawners are solitary males. PMID- 18430147 TI - Glacial vicariance in the Pacific Northwest: evidence from a lodgepole pine mitochondrial DNA minisatellite for multiple genetically distinct and widely separated refugia. AB - The Canadian side of the Pacific Northwest was almost entirely covered by ice during the last glacial maximum, which has induced vicariance and genetic population structure for several plant and animal taxa. Lodgepole pine (Pinus contorta Dougl. ex. Loud.) has a wide latitudinal and longitudinal distribution in the Pacific Northwest. Our main objective was to identify relictual signatures of glacial vicariance in the population structure of the species and search for evidence of distinct glacial refugia in the Pacific Northwest. A maternally inherited mitochondrial DNA minisatellite-like marker was used to decipher haplotype diversity in 91 populations of lodgepole pine located across the natural range. Overall population differentiation was sizeable (G(ST) = 0.365 and R(ST) = 0.568). Four relatively homogeneous groups of populations, possibly representative of as many genetically distinct glacial populations, were identified for the two main subspecies, ssp. latifolia and ssp. contorta. For ssp. contorta, one glacial lineage is suggested to have been located at high latitudes and possibly off the coast of mainland British Columbia (BC), while the other is considered to have been located south of the ice sheet along the Pacific coast. For ssp. latifolia, two genetically distinct glacial populations probably occurred south of the ice sheet: in the area bounded by the Cascades and Rocky Mountains ranges, and on the eastern side of the Rockies. A possible fifth refugium located in the Yukon may have also been present for ssp. latifolia. Zones of contact between these ancestral lineages were also apparent in interior and northern BC. These results indicate the role of the Queen Charlotte Islands and the Alexander Archipelago as a refugial zone for some Pacific Northwest species and the vicariant role played by the Cascades and the American Rocky Mountains during glaciation. PMID- 18430149 TI - Temporal changes in kin structure through a population cycle in a territorial bird, the red grouse Lagopus lagopus scoticus. AB - Populations of red grouse (Lagopus lagopus scoticus) undergo regular multiannual cycles in abundance. The 'kinship hypothesis' posits that such cycles are caused by changes in kin structure among territorial males producing delayed density dependent changes in aggressiveness, which in turn influence recruitment and regulate density. The kinship hypothesis makes several specific predictions about the levels of kinship, aggressiveness and recruitment through a population cycle: (i) kin structure will build up during the increase phase of a cycle, but break down prior to peak density; (ii) kin structure influences aggressiveness, such that there will be a negative relationship between kinship and aggressiveness over the years; (iii) as aggressiveness regulates recruitment and density, there will be a negative relationship between aggressiveness in one year and both recruitment and density in the next; (iv) as kin structure influences recruitment via an affect on aggressiveness, there will be a positive relationship between kinship in one year and recruitment the next. Here we test these predictions through the course of an 8-year cycle in a natural population of red grouse in northeast Scotland, using microsatellite DNA markers to resolve changing patterns of kin structure, and supra-orbital comb height of grouse as an index of aggressiveness. Both kin structure and aggressiveness were dynamic through the course of the cycle, and changing patterns were entirely consistent with the expectations of the kinship hypothesis. Results are discussed in relation to potential drivers of population regulation and implications of dynamic kin structure for population genetics. PMID- 18430150 TI - Catheterization: possible complications and their prevention and treatment. AB - Intermittent catheterization (IC) is a more preferable method for bladder drainage than indwelling urethral or suprapubic catheterization. Several complications with IC have been described, however, including urinary tract infection, genital infection, urethral bleeding, urethritis, urethral stricture, and bladder stones. To prevent these complications, patients should be well instructed on the technique and the risks of IC. Indwelling catheterization should be used only exceptionally, under close control and the catheter should be changed with adequate frequency. PMID- 18430148 TI - Comparative phylogeography of codistributed species of Chilean Liolaemus (Squamata: Tropiduridae) from the central-southern Andean range. AB - In this study, we used a recently developed supertrees method to test for shared phylogeographical signal in partially overlapping geographical ranges of lizards of the genus Liolaemus from the Andean Range in south-central Chile. We reconstruct mtDNA gene trees for three partially codistributed species (Liolaemus tenuis, L. lemniscatus and L. pictus), and our sampling effort is sufficient to allow statistical tests of shared signal between the combinations L. tenuis-L. pictus, and L. tenuis-L. lemniscatus. For both combinations, standardized maximum agreement subtrees scores showed statistically significant signal for shared pattern in regions of overlap, as evaluated by randomization tests (P < 0.001 and < 0.05, respectively). The matrix representation with parsimony tree obtained from the combination of the three different gene trees revealed concordant phylogeographical associations of all species, and was consistent with the geographical association of intraspecific haploclades with three Chilean bioclimatic zones. A multidimensional scaling analysis of several climate variables showed highly significant differences among these zones, which further suggests that they may have contributed to similar patterns of intraspecific divergence across all three species. In the mesomorphic zone in Central Chile, the species L. tenuis and L. lemniscatus may have codiverged in response to shared orogenic vicariant events, which likely predominated over climatic events associated with cycles of glacial advance and retreat. In the hygromorphic zone in southern Chile, however, glacial cycles likely predominated in structuring the phylogeographical histories of L. tenuis and L. pictus, although important ecological differences between these two caution against broad generalizations at this point. PMID- 18430151 TI - The role of testosterone in the pathogenesis of prostate cancer. AB - Relationships between androgenic hormones and prostatic tissue growth are complex. It is certainly true that the prostate will not develop without androgens and the gland will atrophy if androgen support is withdrawn. The hormonal hypothesis remains one of the most important hypotheses in the etiology of prostate cancer (PCa), and efforts are continuing to improve the understanding of androgen actions in PCa. Although evidence from epidemiological studies of associations between circulating levels of androgens and PCa risk has been inconsistent, the traditional view that higher testosterone (T) levels represent a risk factor for PCa appears to have little evidentiary support. Reinvestigation of the relationship between T and PCa seems important and necessary if a new, clinically and scientifically rewarding concept is to be constructed. The present review considers the metabolism and intraprostatic action of T, epidemiological evidence, and the association between T and PCa risk. PMID- 18430152 TI - Relationship between testicular size by ultrasonography and testicular function: measurement of testicular length, width, and depth in patients with infertility. AB - OBJECTIVES: The relationships between simple testicular sizes, including the testicular length, width, and depth and testicular function were evaluated to determine the usefulness of measuring these simple parameters. METHODS: The mean of right and left testicular length, width and depth were measured in 408 men with infertility (mean age, 35.9 +/- 5.3 years) using ultrasonography. The ultrasonographic testicular volumes were calculated as the length x width x depth x 0.71. To evaluate the relationship between the testicular size and function, the patients were divided into seven to ten groups according to each testicular size parameter. RESULTS: The mean testicular volume, length, depth, and width were 13.4 ml, 3.86 cm, 1.80 cm, and 2.59 cm, respectively. These three testicular dimensions significantly correlated with sperm density, total sperm count, total motile sperm count, serum follicle-stimulating hormone and luteinizing hormone equal to ultrasonographic testicular volume. The mean sperm density was in the oligozoospermic range in patients with a mean testicular volume below 10 ml, a mean length below 3.5 cm, a mean depth below 1.75 cm, and a mean width below 2.5 cm. The mean total sperm count was subnormal in patients with a mean testicular volume below 10 ml, a mean length below 3.5 cm, a mean depth below 1.5 cm, and a mean width below 2.25 cm. CONCLUSIONS: The testicular length, width, and depth measured by ultrasonography significantly correlated with the testicular function as well as the ultrasonographic testicular volume. A simple measurement of the testicular length, width, and depth is useful for evaluating the testicular function. PMID- 18430153 TI - Virtual community consultation? Using the literature and weblogs to link community perspectives and health technology assessment. AB - BACKGROUND: Community views, expressed in social impact assessments and collected through community consultation, should play an important role in health technology assessment (HTA). Yet HTA methodologists have been slow to include outcomes of these forms of inquiry in analyses, in part because collecting community views is time-consuming and resource intensive. OBJECTIVE: To explore how community views sourced from published studies, grey literature and informal internet web pages can inform HTA. METHODS: A technology reviewed by Adelaide HTA in 2004 was selected: retinal photography for detection of diabetic retinopathy. Published literature, 'grey' literature and informal web pages were searched to examine the availability of evidence about service community and user community views with respect to this technology. Particular efforts were made to source evidence relating to rural, remote and Aboriginal populations. RESULTS: We found that journal articles, reports from the grey literature and informal internet web pages (including blogs and discussion forums) can provide valuable insight into community views. Although there was little empirical evidence relating to the experience of diabetes and diabetes management in rural, remote and Aboriginal communities, there were indications that some evidence may be transferable from other populations. CONCLUSIONS: Community perspectives on selected health technologies can be gauged from available resources in published and grey literature and perspectives collected in this way can provide insight into whether the introduction of the technology would be acceptable to the community. The limitations of this approach are discussed. PMID- 18430154 TI - Bacterial biomineralization: where to from here? PMID- 18430155 TI - The pedagogical genealogy of Terrance J. Beveridge--a legacy of bonds. AB - Terrance J. Beveridge (TJB) was a professor at the University of Guelph for nearly three decades. He died on September 10, 2007. TJB was a pioneer who pushed the frontier of microbiology and bacteriology. His legacy includes 22 postdoctoral scholars and 24 graduate students. His two dozen graduate student progeny have, in turn, mentored at least 52 additional graduate students. This paper presents TJB's pedagogical tree and touches on many of the topics that he studied during his life including: bacteria cell surfaces, biomineralization, biofilms, and geomicrobiology. PMID- 18430156 TI - Biocontrol strain Pseudomonas fluorescens WCS365 inhibits germination of Fusarium oxysporum spores in tomato root exudate as well as subsequent formation of new spores. AB - Fusarium oxysporum f.sp.radicis-licopersici (Forl) is a soilborne pathogenic fungus which can cause tomato foot and root rot (TFRR). Tomato root exudate is a good source of nutrients for both Forl and the TFRR-suppressing biocontrol bacterium Pseudomonas fluorescens strain WCS365. Incubation of Forl microconidia in tomato root exudate stimulates their germination. This phenomenon is observed, to a lesser extent, upon incubation in plant nutrient solution supplemented with citrate or glucose, the major organic acid and sugar components, respectively, of tomato root exudate. Here we show that induction of germination of microconidia is significantly reduced in the presence of P. fluorescens WCS365 in all tested media. Scanning electron microscopy revealed that P. fluorescens WCS365 colonizes developing hyphae. Efficient colonization correlates with low nutrient availability. Eventually, new microconidia are formed. The presence of P. fluorescens WCS365 reduces the number of newly formed microconidia. This reduction does not depend on physical contact between bacteria and hyphae. We discuss that the ability of P. fluorescens WCS365 to slow down the processes of microconidia germination and development of new microconidia of the phytopathogen, and therefore the ability to reduce fungal dissemination, is likely to contribute to the biocontrol efficacy of this strain. PMID- 18430157 TI - Ciprofloxacin induces mutagenesis to antibiotic resistance independent of UmuC in Streptococcus uberis. AB - Streptococcus uberis is an environmental bovine mastitis pathogen capable of UV inducible SOS mutagenesis. Bacterial SOS systems can be induced by several chemicals including also antibiotics used in clinical practice. Here, we have studied the effect of ciprofloxacin, a fluoroquinolone antibiotic and known inducer of SOS, on mutations leading to antibiotic resistance in S. uberis. Mutation frequencies and spectra were compared in a wild-type S. uberis strain and its DeltaumuC derivative. The results revealed that concentrations of ciprofloxacin corresponding to 0.3-0.5x minimum inhibitory concentration (MIC) induce mutagenesis independent of UmuC. Partial sequencing of the rpoB gene of individual rifampin-resistant clones from wild-type and DeltaumuC strains revealed a similar but complex pattern of point mutations including transitions, transversions and deletions/insertions. It was previously shown that UV induces mainly transition-type mutations and UmuC is essential for the process. Thus, the results presented here demonstrate that S. uberis employs distinct mechanisms for ciprofloxacin and UV-induced mutagenesis, which is a striking difference to Escherichia coli SOS model. PMID- 18430158 TI - Current methodological issues in the study of children with inherited neuromuscular disorders. AB - Several clinical trials assessing children with hereditary neuromuscular disorders have been performed over the last decade. These studies highlighted issues related to design and performance of clinical studies assessing children with this group of disorders. This article reviews recent literature and clinical experience in this area, highlighting methodological shortcomings and disease specific clinical confounding factors that should be considered for future study design. The state of the art of outcome measures in neuromuscular disorders will be discussed, and suggestions for future clinical trials are presented. PMID- 18430159 TI - Roles of Hes genes in neural development. AB - Hes genes are mammalian homologues of Drosophila hairy and Enhancer of split, which encode basic helix-loop-helix (bHLH) transcriptional repressors. In the developing central nervous system, Hes1, Hes3 and Hes5 are highly expressed by neural stem cells. Inactivation of these Hes genes leads to upregulation of proneural genes, acceleration of neurogenesis and premature depletion of neural stem cells. Conversely, overexpression of Hes genes leads to inhibition of neurogenesis and maintenance of neural stem cells. At later stages of development, Hes genes promote gliogenesis. Furthermore, Hes genes regulate maintenance of boundaries, which partition the nervous system into many compartments and endow the neighboring compartments with regional identities by secreting morphogens. Boundary cells usually proliferate slowly and do not give rise to neurons, unlike neural stem cells in compartments. Interestingly, these different characteristics between boundary cells and compartmental neural stem cells are regulated by different modes of Hes1 expression, which is variable in neural stem cells in compartments and persistent and high in boundary cells. Thus, Hes genes play an essential role in neural development by regulating proliferation, differentiation and specification of neural stem cells. PMID- 18430160 TI - Draft genome of the medaka fish: a comprehensive resource for medaka developmental genetics and vertebrate evolutionary biology. AB - The medaka Oryzias latipes is a small egg-laying freshwater teleost, and has become an excellent model system for developmental genetics and evolutionary biology. The medaka genome is relatively small in size, approximately 800 Mb, and the genome sequencing project was recently completed by Japanese research groups, providing a high-quality draft genome sequence of the inbred Hd-rR strain of medaka. In this review, I present an overview of the medaka genome project including genome resources, followed by specific findings obtained with the medaka draft genome. In particular, I focus on the analysis that was done by taking advantage of the medaka system, such as the sex chromosome differentiation and the regional history of medaka species using single nucleotide polymorphisms as genomic markers. PMID- 18430161 TI - Clustered protocadherin family. AB - The brain is a complex system composed of enormous numbers of differentiated neurons, and brain structure and function differs among vertebrates. To examine the molecular mechanisms underlying brain structure and function, it is important to identify the molecules involved in generating neural diversity and organization. The clustered protocadherin (Pcdh) family is the largest subgroup of the diverse cadherin superfamily. The clustered Pcdh proteins are predominantly expressed in the brain and their gene structures in vertebrates are diversified. In mammals, the clustered Pcdh family consists of three gene clusters: Pcdh-alpha, Pcdh-beta, and Pcdh-gamma. During brain development, this family is upregulated by neuronal differentiation, and Pcdh-alpha is then dramatically downregulated by myelination. Clustered Pcdh expression continues in the olfactory bulb, hippocampus, and cerebellum until adulthood. Structural analysis of the first cadherin domain of the Pcdh-alpha protein revealed it lacks the features that classical cadherins require for homophilic adhesiveness, but it contains Pcdh-specific loop structures. In Pcdh-alpha, an RGD motif on a specific loop structure binds beta1-integrin. For gene expression, the gene clusters are regulated by multiple promoters and alternative cis splicing. At the single-cell level, several dozen Pcdh-alpha and -gamma mRNA are regulated monoallelically, resulting in the combinatorial expression of distinct variable exons. The Pcdh alpha and Pcdh-gamma proteins also form oligomers, further increasing the molecular diversity at the cell surface. Thus, the unique features of the clustered Pcdh family may provide the molecular basis for generating individual cellular diversity and the complex neural circuitry of the brain. PMID- 18430162 TI - Elucidation of the role of activin in organogenesis using a multiple organ induction system with amphibian and mouse undifferentiated cells in vitro. AB - Studies performed over the last century have clarified the mechanisms of organ and tissue formation. Mesoderm formation is one of the most important events in early body pattern determination during embryogenesis. In 1988, we found that activin A has mesoderm-inducing activity. As activin A could induce dorsal mesoderm formation, unlike fibroblast growth factor and bone morphogenetic protein, this factor was thought to be the molecular entity of the Spemann Mangold organizer. Subsequently, the mechanisms of early embryogenesis have been clarified using molecular biological techniques, resulting in the identification of many genes that are involved in organ and tissue development. This finding that activin A could induce dorsal mesoderm formation spurred research into the application of agents that induce organs and tissues in vitro. In this regard, we have shown that many organ types can be induced by activin A in vitro. Moreover, we have found that other types of organs can be induced by changing the conditions of treatment. To date, more than 20 different types of tissues and organs have been successfully induced from Xenopus undifferentiated cells in vitro. In recent years, we have applied these protocols to mouse embryonic stem cells, and we have successfully induced several tissues, such as the pancreas and cardiomyocytes. We are also investigating how the pluripotency of undifferentiated stem cells is regulated. In this review, we summarize the current knowledge regarding activin as a mesoderm-inducing factor and its application for the induction of tissues and organs from undifferentiated cells. Moreover, we provide some examples of in vitro tissue differentiation from mouse embryonic stem cells, which may prove useful in regenerative medicine. PMID- 18430163 TI - Developmental patterning deciphered in avian chimeras. AB - I started my scientific carer by investigating the development of the digestive tract in the laboratory of a well-known embryologist, Etienne Wolff, then professor at the College de France. My animal model was the chick embryo. The investigations that I pursued on liver development together with serendipity, led me to devise a cell-marking technique based on the construction of chimeric embryos between two closely related species of birds, the Japanese quail (Coturnix coturnix japonica) and the chick (Gallus gallus). The possibility to follow the migration and fate of the cells throughout development from early embryonic stages up to hatching and even after birth, was a breakthrough in developmental biology of higher vertebrates. This article describes some of scientific achievements based on the use of this technique in my laboratory during the last 38 years. PMID- 18430164 TI - Evolutionary developmental studies of cyclostomes and the origin of the vertebrate neck. AB - Because they lack some gnathostome-specific traits, cyclostomes have often been regarded as representing an intermediate state linking non-vertebrate chordates and gnathostomes. To understand the evolutionary origins of the jaw and paired fins, lamprey embryos and larvae have been used as comparative models. The lack of the jaw-neck region is a conspicuous feature specific to cyclostomes; however, the absence of these features has been largely neglected both in evolutionary developmental studies and in the field of classical comparative embryology. This review seeks to develop a possible evolutionary scenario of the vertebrate neck muscles by taking the cucullaris (trapezius) muscle as the focus. By combining the comparative embryology of lampreys and gnathostomes, and considering the molecular-level developmental mechanism of skeletal muscle differentiation, this review argues that the establishment of the vertebrate neck deserves to be called an evolutionary novelty based on the remodeling of mesenchymal components between the cranium and the shoulder girdle, which involves both mesodermal and neural crest cell lineages. PMID- 18430165 TI - Molecular analysis of endoderm regionalization. AB - We have engaged in a number of studies in our laboratory that have focused on the molecular mechanisms underlying gut formation, with particular attention being paid to the establishment of regional differences found in the entire gut and within each digestive organ. We have found from our analyses that the presumptive fate of the endoderm in the embryos of vertebrates is determined quite early during development, but the realization of this fate often requires molecular cues from the neighboring tissues such as the lateral plate mesoderm and the mesenchyme derived from it. The mesenchyme seems often to exert instructive or supportive induction effects and, in some cases, a completely inhibitory role during the differentiation of the endodermal epithelium. In addition, many reports on the formation of the stomach, intestine, liver and salivary gland in vertebrates, and of Drosophila gut, all indicate that the morphogenesis and cytodifferentiation of these organs are regulated by the regulated expression of genes encoding growth factors and transcription factors. We have further shown that the epithelium can regulate the differentiation of the mesenchyme into the connective tissue and the smooth muscle layers, thus demonstrating the occurrence of literally interactive processes in the development of the digestive organs. PMID- 18430166 TI - Sexual development of mouse germ cells: Nanos2 promotes the male germ cell fate by suppressing the female pathway. AB - Much research has been conducted in recent years to elucidate the mechanisms underlying the crucial developmental process of sex determination. It has now been shown that somatic sex is principally determined by the chromosomal sex and the molecular mechanisms involved in this process have become relatively well understood in both human and mouse. However, the pathways involved in the sex determination of the germ cells remain largely unknown except for the fact that the somatic cues surrounding these cells play a significant role. Moreover, which sexual pathway of the germ cells is induced or suppressed has long been a subject of some dispute. Recent findings indicate that the key molecule that influences this choice is retinoic acid. In addition, the Nanos protein has been shown to play a critical role in promoting male germ cell differentiation. In this review, the possible mechanisms underlying these events, which have been brought to light by recent findings, are summarized to provide a better and more precise understanding of our current knowledge of the sex determination and subsequent differentiation of germ cells. PMID- 18430167 TI - Shedding light on developmental gene regulation through the lens. AB - My group has long studied transcriptional gene regulation involved in cell differentiation, employing lens cell differentiation as a model. In this article, our progress over the last quarter of a century in deciphering the principles involved in developmental gene regulation is described, outlining concurrent advancement in relevant branches of developmental biology. PMID- 18430168 TI - Retrospective on reverse genetics in mice around the world and in Japan. AB - The 2007 Nobel Prize for Physiology or Medicine was awarded to Mario R. Capecchi, Martin J. Evans and Oliver Smithies for their contribution in generating mutant mice by gene targeting in embryonic stem (ES) cells. Although there are many experimental animals, it is yet only in mouse that one can genetically examine functions of genes at will. It was merely a dream in the early 1980s that genetic studies with mutants would one day become a reality in mammals. The story began with tetratocarcinoma/embryonal carcinoma cells. Now, through the successes of cloning in mammals, somatic cells such as our skin cells will shortly be transformed into ES-like (induced pluripotent stem) cells by the proper activation of endogenous genes such as Oct4 and Sox2 with chemicals. How have times changed? PMID- 18430169 TI - Transgenic technology for visualization and manipulation of the neural circuits controlling behavior in zebrafish. AB - The vertebrate brain is innately equipped with neural circuits that make quick behavioral decisions possible. Elucidating these neural circuits, determining how their master plans are encoded in the genome, and revealing how they can be modified by postnatal experiences will facilitate our understanding of how nature and nurture interact to establish an animal's behavior. In this review, we explain how transgenic zebrafish can cast insights into the developmental mechanisms and functional roles of the neural circuits that directly and indirectly control visuomotor behavior, by taking as an example a transgenic line Tg(brn3a-hsp70:GFP) enabling visualization of the tectobulbar and habenulo interpeduncular tracts. These insights emphasize the benefits of applying advanced transgenic technology in zebrafish to future research into this area. PMID- 18430170 TI - Cadherins in neuronal morphogenesis and function. AB - Classic cadherins represent a family of calcium-dependent homophilic cell-cell adhesion molecules. They confer strong adhesiveness to animal cells when they are anchored to the actin cytoskeleton via their cytoplasmic binding partners, catenins. The cadherin/catenin adhesion system plays key roles in the morphogenesis and function of the vertebrate and invertebrate nervous systems. In early vertebrate development, cadherins are involved in multiple events of brain morphogenesis including the formation and maintenance of the neuroepithelium, neurite extension and migration of neuronal cells. In the invertebrate nervous system, classic cadherin-mediated cell-cell interaction plays important roles in wiring among neurons. For synaptogenesis, the cadherin/catenin system not only stabilizes cell-cell contacts at excitatory synapses but also assembles synaptic molecules at synaptic sites. Furthermore, this system is involved in synaptic plasticity. Recent studies on the role of individual cadherin subtypes at synapses indicate that individual cadherin subtypes play their own unique role to regulate synaptic activities. PMID- 18430171 TI - Interview with Dr Tokindo S. Okada in commemoration of the 50th volume of Development, Growth and Differentiation. Interview by Yumi Momiki, Elizabeth Nakajima, Kiyokazu Agata. PMID- 18430172 TI - Comparative evaluation of beneficiary effects of priming agents (2% hydroquinone and 0.025% retinoic acid) in the treatment of melasma with glycolic acid peels. AB - BACKGROUND: Chemical peels have become a popular modality in the treatment of melasma. The most disturbing side effect of this procedure is postinflammatory hyperpigmentation. This may be minimized with the help of priming agents. Because there is a paucity of such studies, this study was taken up to evaluate the beneficial effects of hydroquinone versus tretinoin as priming agents in treatment of melasma with glycolic acid peels. METHODS: Sixty patients of melasma were randomly assigned in three groups of 20 patients each in a single-blind study. Group I received only glycolic acid peels while Groups II and III were primed with 0.025% tretinoin and 2% hydroquinone, respectively, for 2 weeks before peeling. The patients received serial glycolic acid peels fortnightly for the first 3 months and then monthly for the next 3 months and were then followed up for the next 3 months when peeling was stopped. Clinical and photographic evaluation was done at 3, 6, and 9 months, and subjective improvement was noted. RESULTS: There was an overall decrease in MASI from baseline to 6 months in all three groups but it was highly significant between Groups I and III (p<.001) at 6 and 9 months and significant between Groups II and III (p<.01) at 9 months. CONCLUSIONS: Results are better with hydroquinone as priming agent compared to tretinoin in enhancing the results with glycolic acid peels in melasma and in decreasing postpeel postinflammatory hyperpigmentation. PMID- 18430173 TI - A double-blind, comparative study of nonanimal-stabilized hyaluronic acid versus human collagen for tissue augmentation of the dorsal hands. AB - BACKGROUND AND OBJECTIVE: Cosmetic surgery to counteract the aging process is an evolving field. Most procedures have concentrated on the face; however, the hands are an often-neglected area. Current methods of hand rejuvenation include autologous fat injection, sclerotherapy, intense pulsed light, chemical peel, and microdermabrasion. Only autologous fat injection restores dermal thinning. We compare the use of hyaluronic acid (Restylane, Medicis Aesthetics Inc.) versus collagen (Cosmoplast, INAMED Aesthetics) for soft tissue augmentation of the dorsal hands. MATERIALS AND METHODS: Ten female patients who demonstrated dermal thinning of the dorsal hands were randomized to receive 1.4 mL of hyaluronic acid or 2.0 cm(3) collagen to alternate interphalangeal spaces of dorsal hands. Patients returned at 1 week, 1 month, 3 months, and 6 months for digital photography and completion of a patient/physician questionnaire. RESULTS: Hands were scored by two separate blinded physicians on scales of 1 to 5 for clearance of veins. Patients scored both tolerability and satisfaction on a scale of 1 to 5. Analysis showed a mean difference of 0.95 (0.004), median difference of 0.9 (0.008) for clearance, and a mean difference of 0.90 (0.010) with a median difference of 1.0 (0.031). The satisfaction difference was not significant with a mean difference of 0.80 (0.070) and median difference of 1.0 (0.117). CONCLUSION: Aging of the hands is a common problem that is often overlooked. The use of soft tissue fillers is a viable tool in hand rejuvenation. In this study hyaluronic acid proved to be superior in efficacy to collagen. PMID- 18430174 TI - Effect of quilting sutures on hematoma formation after liposuction with dermal curettage for treatment of axillary hyperhidrosis: a randomized clinical trial. AB - BACKGROUND: Liposuction provides further reduction of axillary sweating and malodor when combined with dermal curettage with sharp rasping cannulas. This aggressive approach is associated with relatively higher rate of hematoma formation when compared to the conventional simple liposuction. OBJECTIVE: The aim of this prospective, randomized, controlled trial was to evaluate the effect of quilting sutures on the incidence of hematoma formation after liposuction curettage for treatment of axillary hyperhidrosis (AH). MATERIALS AND METHODS: The trial randomized 59 male patients (118 axillae) undergoing liposuction curettage for AH to quilting procedures (61 axillae) or control group (57 axillae) for intention-to-treat analysis. Outcome measures included the incidence of hematoma formation, operative time, degree of postoperative pain, and amount of analgesics consumption. RESULTS: Quilting sutures significantly reduced the incidence of axillary hematoma from 28.1% to 4.9%. Quilting was associated with the lengthening of operative time but did not affect the postoperative pain. CONCLUSION: Considering its efficacy in reducing postoperative hematoma, quilting is recommended in combination with aggressive liposuction-curettage procedure for treating AH and osmidrosis. PMID- 18430175 TI - Histological and clinical findings in different surgical strategies for focal axillary hyperhidrosis. AB - INTRODUCTION: Although a variety of different surgical strategies for focal axillary hyperhidrosis (FAH) have proven effective, little is known of intraoperative and postoperative histologies of different surgical methods. OBJECTIVE: The objective was to use pre-, intra-, and postoperative histologic findings to evaluate different surgical procedures for FAH in establishing a possible correlation between the interventions and clinical outcome. MATERIAL AND METHODS: A total of 40 patients underwent surgery with 15 undergoing liposuction curettage (LC), 14 radical skin excision (RSE) with Y-plasty closure, and 11 a skin-sparing technique (SST). Before surgery, density and ratio of eccrine and apocrine sweat glands were evaluated with routine histology. Further biopsies were taken directly after surgery in the RSE and SST groups and 1 year postoperatively in all patients. Additionally, gravimetry was performed, side effects were documented, and patients were asked to evaluate the aesthetic outcome of the surgical method by using an analogue scale. RESULTS: Preoperatively, the mean density of eccrine glands was 11.1/cm(2) compared to 16.9/cm(2) apocrine glands (apocrine/eccrine ratio, 1.6). Biopsy specimen directly after surgery showed remaining sweat glands in 7/15 (46.7%) LC patients and in 4/11 (36.4%) of the SST patients. One year after surgery, sweat gland density was significantly reduced in the LC (79.1%) and the SST (74.9%) groups. In the RSE group, only scar formation was present. Gravimetry showed significantly reduced sweat rates 12 months after surgery in all groups (LC, 66.4%; SST, 62.9%; RSE, 65.3% [p<.05]). Most frequent side effects were hematoma (LC, n=3; SST, n=2; RSE, n=3), subcutaneous fibrotic bridles (LC, n=8; SST, n=3; RSE, n=0), skin erosion (LC, n=3; SST, n=4; RSE, n=0), focal hair loss (LC, n=9; SST, n=11; RSE, n=14), and paresthesia (LC, n=4; SST, n=3; RSE, n=5). CONCLUSION: Histologic distribution and density of sweat glands were comparable to previous studies. All three surgical procedures evaluated are effective in the treatment of FAH. RSE and SST techniques are associated with a higher risk of side effects and cause more extensive scarring. However, one LC patient (n=1; 6.7%) did not respond to treatment. PMID- 18430176 TI - Efficacy of low-level laser therapy for chronic cutaneous ulceration in humans: a review and discussion. AB - Chronic wounds, particularly venous ulcerations, are notoriously difficult to heal. Because current therapies are variable in their ability to induce complete healing, there remains a need to develop adjunctive treatments that can improve or accelerate the healing process. The use of low-energy lasers to stimulate wound healing has been pursued over many decades in studies of varying quality. This form of treatment has had high appeal due to its novelty, relative ease, and low morbidity profile. The authors reviewed the available published literature on low-level laser technology in an attempt to provide cumulative insight on the effect of this treatment for wound healing. PMID- 18430177 TI - Selective dose escalation of chemoradiotherapy for locally advanced esophageal cancer. AB - This phase II study assessed the use of concurrent continuous infusion of 5 fluorouracil and weekly carboplatin plus paclitaxel with selective radiation dose escalation for patients with localized esophageal cancer. Patients with esophageal carcinoma were staged by thoracic and abdominal computed tomography, endoscopic ultrasound, and positron emission tomography scans. Patients received a continuous infusion of 5-fluorouracil 225 mg/m(2) on days 1 to 38 and intravenous paclitaxel 45 mg/m(2) and carboplatin AUC 2 on days 1, 8, 15, 22, 29, and 36. Radiotherapy was delivered in 1.8-Gy fractions, 5 d/wk for 5.5 weeks. Six to 8 weeks after initial therapy, patients without metastatic progression but with a positive biopsy, or less than partial response received a 9-Gy boost with the same concurrent chemotherapy. Twenty-four patients were enrolled: 18 patients were enrolled initially; 6 additional patients were enrolled following a protocol amendment designed to reduce the esophagitis by adding the radioprotectant amifostine. Median follow-up was 30 months. Twenty (83%) patients had adenocarcinomas of the lower esophagus/gastroesophageal junction. Seventeen patients (81%) attained at least a partial response. Six patients received boost treatment. At 4 years, overall survival was 28%, cause-specific survival was 38%, locoregional control was 61%, and distant metastasis-free survival was 52%. Radiation delays ranged from 0 to 62 days (median, 8 d), primarily owing to esophagitis. In total, 28% of patients developed esophageal strictures requiring dilatations. There were no differences in esophageal strictures, local control, or survival with the addition of amifostine. PMID- 18430178 TI - Chemosensitivity of patients with recurrent esophageal cancer receiving perioperative chemotherapy. AB - Perioperative chemotherapy (CT) and chemoradiotherapy are widely used for advanced esophageal cancer. We evaluated the chemosensitivity of patients displaying recurrent esophageal cancer after esophagectomy with perioperative CT. From the database at National Cancer Center Hospital in Tokyo, we extracted recurrent esophageal cancer cases after perioperative CT and evaluated the effectiveness of the first CT against the recurrent disease according to the duration between termination of the original perioperative CT and recurrence with treatment-free intervals (TFIs) 6 months. Systemic CT for their recurrent disease was performed for 30 esophageal cancer patients after perioperative CT. All patients received 5-fluorouracil and cisplatin as perioperative CT, with relapses occurring at TFIs 6 months in 19 patients (all received platinum containing regimens). The response rate of patients experiencing a recurrence at TFIs 6 months was 0 and 37% (P = 0.029), the median progression-free survival was 2.8 and 4.8 months (log-rank P = 0.001) and the median overall survival was 6.1 and 10.2 months (log-rank P = 0.012), respectively. Recurrence at the TFI 5.5 mm. There was no difference in median number of bougienage procedures in these subgroups (4.5 [2-9] vs. 4 [2 20]). Patients who presented with BAS earlier than 6 weeks postoperative had more procedures (median 8 [2-20] vs. 4 [2-9]) than those presenting later. Patients in whom first bougienage was possible to only 16 mm diameter needed more procedures than patients in whom first dilation was possible to more than 16 mm (median 5.5 [3-20] vs. 3 [2-9]). In conclusion, both early BAS development and the diameter of bougienage at first endoscopy, but not the extent of stricture, seem to be predictive factors for longer bougienage therapy. In order to influence the BAS formation early, we now routinely examine every patient after esophageal resection endoscopically in the 6th postoperative week. PMID- 18430181 TI - Utility of esophageal impedance in identifying dysmotility in patients with erosive esophagitis. AB - SUMMARY: This study was intended to assess the utility of combined multiple intraluminal impedance and esophageal manometry (MII-EM) in evaluating reflux patients and in identifying those with esophageal dysmotility. Thirteen controls and 20 patients with gastroesophageal reflux disease (GERD) underwent combined MII-EM with a catheter containing four impedance-measuring segments and four solid-state pressure transducers. Each subject received 10 liquid and 10 viscous boluses to be swallowed. Distal esophageal contraction amplitude was significantly lower in GERD patients than in controls for viscous swallows (58.3 +/- 7.3 mmHg versus 82.4 +/- 4.1 mmHg, P = 0.005). Total bolus transit time was significantly slower in GERD patients than in controls for liquid swallows (P = 0.035). The percentages of complete bolus transit were significantly lower in GERD patients compared with controls (all P = 0.005). Half of GERD patients with normal EM still had abnormal bolus transit while three-quarters of those with abnormal EM had abnormal bolus transit. MII helps identify bolus transit abnormalities not detected by conventional manometry. Combined MII-EM is clinically useful for detecting esophageal dysmotility in patients with erosive esophagitis. PMID- 18430182 TI - Clinicopathologic and prognostic factors of young and elderly patients with esophageal adenocarcinoma: is there really a difference? AB - Evidence suggests a significant difference in the incidence, presentation, and outcome of young and elderly patients with esophageal adenocarcinoma. We aimed to compare clinicopathologic and prognostic factors of young and elderly patients with esophageal adenocarcinoma at a surgical department in Europe. From 1996 to 2006, 223 patients with a resectable esophageal adenocarcinoma were analyzed and divided in three groups: (i) patients 70 years (n = 52). Clinicopathological and prognostic factors were compared between these groups. The total number of patients with esophageal adenocarcinoma increased significantly. Although the total number of patients 0.20) and the adjusted aROC was 0.920 for 5 years of follow-up. CONCLUSION: The risk score had adequate performance. Further validations in other cohorts of patients with T2DM are needed before clinical use. PMID- 18430206 TI - Reasons for not using smoking cessation aids. AB - BACKGROUND: Few smokers use effective smoking cessation aids (SCA) when trying to stop smoking. Little is known why available SCA are used insufficiently. We therefore investigated the reasons for not using SCA and examined related demographic, smoking behaviour, and motivational variables. METHODS: Data were collected in two population-based studies testing smoking cessation interventions in north-eastern Germany. A total of 636 current smokers who had never used SCA and had attempted to quit or reduce smoking within the last 12 months were given a questionnaire to assess reasons for non-use. The questionnaire comprised two subscales: "Social and environmental barriers" and "SCA unnecessary." RESULTS: The most endorsed reasons for non-use of SCA were the belief to be able to quit on one's own (55.2%), the belief that help is not necessary (40.1%), and the belief that smoking does not constitute a big problem in one's life (36.5%). One quarter of all smokers reported that smoking cessation aids are not helpful in quitting and that the aids cost too much. Smokers intending to quit agreed stronger to both subscales and smokers with lower education agreed stronger to the subscale "Social and environmental barriers". CONCLUSION: Main reasons for non-use of SCA are being overly self-confident and the perception that SCA are not helpful. Future interventions to increase the use of SCA should address these reasons in all smokers. PMID- 18430207 TI - Self-reported work ability of Norwegian women in relation to physical and mental health, and to the work environment. AB - OBJECTIVES: To examine the self-reported level of work ability among female employees and the relationship between work ability and demographic characteristics, physical health, mental health, and various psychosocial and organizational work environment factors. METHODS: Participants were 597 female employees with an average age of 43 years from urban and rural areas in Norway. Trained personnel performed a structured interview to measure demographic variables, physical health, and characteristics of the working environment. Mental health was assessed using the 25-item version of the Hopkins Symptoms Checklist (HSCL-25). Work ability was assessed using a question from the Graded Reduced Work Ability Scale. RESULTS: Of the 597 female employees, 8.9% reported an extremely or very reduced ability to work. Twenty-four percent reported poor physical health and 21.9% reported mental distress (>/= 1.55 HSCL-25 cut-off). Women, who reported moderately and severely reduced work ability, did not differ a lot. Moderately reduced work ability increased with age and was associated with physical and mental health. Severely reduced work ability was strongly associated only with physical health and with unskilled occupation. Of eight work environment variables, only three yielded significant associations with work ability, and these associations disappeared after adjustment in the multivariate analysis. CONCLUSION: Results indicate that ageing, in addition to poor self reported physical health and unskilled work, were the strongest factors associated with reduced work ability among female employees. Impact of work environment in general was visible only in univariate analysis. PMID- 18430208 TI - Quantum dots in axillary lymph node mapping: biodistribution study in healthy mice. AB - BACKGROUND: Breast cancer is the first cause of cancer death among women and its incidence doubled in the last two decades. Several approaches for the treatment of these cancers have been developed. The axillary lymph node dissection (ALND) leads to numerous morbidity complications and is now advantageously replaced by the dissection and the biopsy of the sentinel lymph node. Although this approach has strong advantages, it has its own limitations which are manipulation of radioactive products and possible anaphylactic reactions to the dye. As recently proposed, these limitations could in principle be by-passed if semiconductor nanoparticles (quantum dots or QDs) were used as fluorescent contrast agents for the in vivo imaging of SLN. QDs are fluorescent nanoparticles with unique optical properties like strong resistance to photobleaching, size dependent emission wavelength, large molar extinction coefficient, and good quantum yield. METHODS: CdSe/ZnS core/shell QDs emitting around 655 nm were used in our studies. 20 microL of 1 microM (20 pmol) QDs solution were injected subcutaneously in the anterior paw of healthy nude mice and the axillary lymph node (ALN) was identified visually after injection of a blue dye. In vivo fluorescence spectroscopy was performed on ALN before the mice were sacrificed at 5, 15, 30, 60 min and 24 h after QDs injection. ALN and all other organs were removed, cryosectioned and observed in fluorescence microscopy. The organs were then chemically made soluble to extract QDs. Plasmatic, urinary and fecal fluorescence levels were measured. RESULTS: QDs were detected in ALN as soon as 5 min and up to 24 h after the injection. The maximum amount of QDs in the ALN was detected 60 min after the injection and corresponds to 2.42% of the injected dose. Most of the injected QDs remained at the injection site. No QDs were detected in other tissues, plasma, urine and feces. CONCLUSION: Effective and rapid (few minutes) detection of sentinel lymph node using fluorescent imaging of quantum dots was demonstrated. This work was done using very low doses of injected QDs and the detection was done using a minimally invasive method. PMID- 18430209 TI - Development of a triage protocol for patients presenting with gastrointestinal hemorrhage: a prospective cohort study. AB - INTRODUCTION: Many patients presenting with acute gastrointestinal hemorrhage (GIH) are admitted to the intensive care unit (ICU) for monitoring. A simple triage protocol based upon validated risk factors could decrease ICU utilization. METHODS: Records of 188 patients admitted with GIH from the emergency department (ED) were reviewed for BLEED criteria (visualized red blood, systolic blood pressure below 100 mm Hg, elevated prothrombin time [PT], erratic mental status, and unstable comorbid disease) and complication within the first 24 hours of admission. Variables associated with early complication were reassessed in 132 patients prospectively enrolled as a validation cohort. A triage model was developed using significant predictors. RESULTS: We studied 188 patients in the development set and 132 in the validation set. Red blood (relative risk [RR] 4.53, 95% confidence interval [CI] 2.04, 10.07) and elevated PT (RR 3.27, 95% CI 1.53, 7.01) were significantly associated with complication in the development set. In the validation cohort, the combination of red blood or unstable comorbidity had a sensitivity of 0.73, a specificity of 0.55, a positive predictive value of 0.24, and a negative predictive value of 0.91 for complication within 24 hours. In simulation studies, a triage model using these variables could reduce ICU admissions without increasing the number of complications. CONCLUSION: Patients presenting to the ED with GIH who have no evidence of ongoing bleeding or unstable comorbidities are at low risk for complication during hospital admission. A triage model based on these variables should be tested prospectively to optimize critical care resource utilization in this common condition. PMID- 18430210 TI - Efficacy and safety of a subacromial continuous ropivacaine infusion for post operative pain management following arthroscopic rotator cuff surgery: a protocol for a randomised double-blind placebo-controlled trial. AB - BACKGROUND: Major shoulder surgery often results in severe post-operative pain and a variety of interventions have been developed in an attempt to address this. The continuous slow infusion of a local anaesthetic directly into the operative site has recently gained popularity but it is expensive and as yet there is little conclusive evidence that it provides additional benefits over other methods of post-operative pain management. METHODS/DESIGN: This will be a randomised, placebo-controlled trial involving 158 participants. Following diagnostic arthroscopy, all participants will undergo arthroscopic subacromial decompression with or without rotator cuff repair, all operations performed by a single surgeon. Participants, the surgeon, nurses caring for the patients and outcome assessors will be blinded to treatment allocation. All participants will receive a pre-incision bolus injection of 20 mls of ropivacaine 1% into the shoulder and an intra-operative intravenous bolus of parecoxib 40 mg. Using concealed allocation participants will be randomly assigned to active treatment (local anaesthetic ropivacaine 0.75%) or placebo (normal saline) administered continuously into the subacromial space by an elastomeric pump at 5 mls per hour post-operatively. Patient controlled opioid analgesia and oral analgesics will be available for breakthrough pain. Outcome assessment will be at 15, 30 and 60 minutes, 2, 4, 8, 12, 18 and 24 hours, and 2 or 4 months for decompression or decompression plus repair respectively. The primary end point will be average pain at rest over the first 12-hour post-operative period on a verbal analogue pain score. Secondary end points will be average pain at rest over the second 12 hour post-operative period, maximal pain at rest over the first and second 12 hour periods, amount of rescue medication used, length of inpatient stay and incidence of post-operative adhesive capsulitis. DISCUSSION: The results of this trial will contribute to evidence-based recommendations for the effectiveness of pain management modalities following arthroscopic rotator cuff surgery. If the local anaesthetic pain-buster provides no additional benefits over placebo then valuable resources can be put to better use in other ways. TRIAL REGISTRATION: Australian Clinical Trials Register Number ACTR12606000195550. PMID- 18430211 TI - Regulation of 5'-adenosine monophosphate deaminase in the freeze tolerant wood frog, Rana sylvatica. AB - BACKGROUND: The wood frog, Rana sylvatica, is one of a few vertebrate species that have developed natural freeze tolerance, surviving days or weeks with 65-70% of its total body water frozen in extracellular ice masses. Frozen frogs exhibit no vital signs and their organs must endure multiple stresses, particularly long term anoxia and ischemia. Maintenance of cellular energy supply is critical to viability in the frozen state and in skeletal muscle, AMP deaminase (AMPD) plays a key role in stabilizing cellular energetics. The present study investigated AMPD control in wood frog muscle. RESULTS: Wood frog AMPD was subject to multiple regulatory controls: binding to subcellular structures, protein phosphorylation, and effects of allosteric effectors, cryoprotectants and temperature. The percentage of bound AMPD activity increased from 20 to 35% with the transition to the frozen state. Bound AMPD showed altered kinetic parameters compared with the free enzyme (S0.5 AMP was reduced, Hill coefficient fell to approximately 1.0) and the transition to the frozen state led to a 3-fold increase in S0.5 AMP of the bound enzyme. AMPD was a target of protein phosphorylation. Bound AMPD from control frogs proved to be a low phosphate form with a low S0.5 AMP and was phosphorylated in incubations that stimulated PKA, PKC, CaMK, or AMPK. Bound AMPD from frozen frogs was a high phosphate form with a high S0.5 AMP that was reduced under incubation conditions that stimulated protein phosphatases. Frog muscle AMPD was activated by Mg.ATP and Mg.ADP and inhibited by Mg.GTP, KCl, NaCl and NH4Cl. The enzyme product, IMP, uniquely inhibited only the bound (phosphorylated) enzyme from muscle of frozen frogs. Activators and inhibitors differentially affected the free versus bound enzyme. S0.5 AMP of bound AMPD was also differentially affected by high versus low assay temperature (25 vs 5 degrees C) and by the presence/absence of the natural cryoprotectant (250 mM glucose) that accumulates during freezing. CONCLUSION: Maintenance of long term viability under the ischemic conditions in frozen muscle requires attention to the control of cellular energetics. Differential regulatory controls on AMPD by mechanisms including binding to muscle proteins, actions allosteric effectors, glucose and temperature effects and reversible phosphorylation adjust enzyme function for an optimal role in controlling cellular adenylate levels in ischemic frozen muscle. Stable modification of AMPD properties via freeze-responsive phosphorylation may contribute both to AMPD control and to coordinating AMPD function with other enzymes of energy metabolism in cold ischemic muscle. PMID- 18430212 TI - Heritability of cardiovascular risk factors in a Brazilian population: Baependi Heart Study. AB - BACKGROUND: The heritability of cardiovascular risk factors is expected to differ between populations because of the different distribution of environmental risk factors, as well as the genetic make-up of different human populations. METHODS: The purpose of this analysis was to evaluate genetic and environmental influences on cardiovascular risk factor traits, using a variance component approach, by estimating the heritability of these traits in a sample of 1,666 individuals in 81 families ascertained randomly from a highly admixed population of a city in a rural area in Brazil. RESULTS: Before adjustment for sex, age, age2, and age x sex interaction, polygenic heritability of systolic (SBP) and diastolic (DBP) blood pressure were 15.0% and 16.4%, waist circumference 26.1%, triglycerides 25.7%, fasting glucose 32.8%, HDL-c 31.2%, total cholesterol 28.6%, LDL-c 26.3%, BMI 39.1%. Adjustment for covariates increased polygenic heritability estimates for all traits mainly systolic and diastolic blood pressure (25.9 and 26.2%, respectively), waist circumference (40.1%), and BMI (51.0%). CONCLUSION: Heritability estimates for cardiovascular traits in the Brazilian population are high and not significantly different from other studied worldwide populations. Mapping efforts to identify genetic loci associated with variability of these traits are warranted. PMID- 18430213 TI - Characterisation of the tryptophan synthase alpha subunit in maize. AB - BACKGROUND: In bacteria, such as Salmonella typhimurium, tryptophan is synthesized from indole-3-glycerole phosphate (IGP) by a tryptophan synthase alphabetabetaalpha heterotetramer. Plants have evolved multiple alpha (TSA) and beta (TSB) homologs, which have probably diverged in biological function and their ability of subunit interaction. There is some evidence for a tryptophan synthase (TS) complex in Arabidopsis. On the other hand maize (Zea mays) expresses the TSA-homologs BX1 and IGL that efficiently cleave IGP, independent of interaction with TSB. RESULTS: In order to clarify, how tryptophan is synthesized in maize, two TSA homologs, hitherto uncharacterized ZmTSA and ZmTSAlike, were functionally analyzed. ZmTSA is localized in plastids, the major site of tryptophan biosynthesis in plants. It catalyzes the tryptophan synthase alpha-reaction (cleavage of IGP), and forms a tryptophan synthase complex with ZmTSB1 in vitro. The catalytic efficiency of the alpha-reaction is strongly enhanced upon complex formation. A 160 kD tryptophan synthase complex was partially purified from maize leaves and ZmTSA was identified as native alpha subunit of this complex by mass spectrometry. ZmTSAlike, for which no in vitro activity was detected, is localized in the cytosol. ZmTSAlike, BX1, and IGL were not detectable in the native tryptophan synthase complex in leaves. CONCLUSION: It was demonstrated in vivo and in vitro that maize forms a tryptophan synthase complex and ZmTSA functions as alpha-subunit in this complex. PMID- 18430214 TI - A Markov model to evaluate hospital readmission. AB - BACKGROUND: The analysis of non-fatal recurring events is frequently found in studies on chronic-degenerative diseases. The aim of this paper is to estimate the probability of readmission of patients with Chronic Obstructive Pulmonary Disease (COPD) or with Respiratory Failure (RF). METHODS: The Repeated hospital admissions of a patient are considered as a Markov Chain. The transitions between the states are estimated using the Nelson-Aalen estimator. The analysis was carried out using the Puglia Region hospital patient discharge database for the years 1998-2005. Patients were selected on the basis of first admission between 01/01/2001 and 31/12/2005 with ICD-9-CM code of COPD or RF as principal and/or secondary diagnosis. For those selected two possible transitions were considered in the case they had the second and third admission with an ICD-9-CM code of COPD or RF as principal diagnosis. RESULTS: The probability of readmission is increased in patients with a diagnosis of RF (OR = 1.618 in the first transition and 1.279 in the second) and also in those with a diagnosis of COPD or RF as the principal diagnosis at first admission (OR = 1.615 in the first transition and 1.193 in the second). The clinical gravity and the ward from which they were discharged did not significantly influence the probability of readmission. CONCLUSION: The time to readmission depends on the gravity of the pathology at onset. In patients with a grave clinical picture, either COPD or Respiratory Failure, when treated and controlled after the first admission, they become minor problems and they are indicated among secondary diagnoses in any further admission. PMID- 18430215 TI - Drotrecogin alfa (activated): real-life use and outcomes for the UK. AB - INTRODUCTION: In March 2001, the results of the Recombinant Human Activated Protein C Worldwide Evaluation in Severe Sepsis (PROWESS) study were published, which indicated a 6.1% absolute reduction in 28-day mortality. Drotrecogin alfa (activated; DrotAA) was subsequently approved for use in patients with severe sepsis. METHODS: In December 2002, critical care units in England, Wales and Northern Ireland were invited to participate in an audit of DrotAA. Data for each infusion of DrotAA were linked to case mix and outcome data from a national audit. Use of DrotAA was described and a nonrandomized comparison of effectiveness was conducted. RESULTS: 1,292 infusions of DrotAA were recorded in 112 units; 61% commenced during the first 24 hours in the unit. The majority (77%) of patients had three or more organs failing; lung (42%) and abdomen (40%) were the most common primary sites of infection. Crude hospital mortality was high (45%); at 28 days, only 18% had left acute hospital and 19% were still in the unit. For 30%, the full 96-hour infusion was not completed; 24% of infusions were interrupted; 8.1% experienced one or more serious adverse events, of which 77% were serious bleeding events. Of eight relative risks estimated from individually-matched (0.75 to 0.85) and propensity-matched (0.82 to 0.90) controls, seven were consistent with the results of PROWESS. Restricting the analysis to patients receiving DrotAA during the first 24 hours resulted in larger treatment effects (relative risks 0.62 to 0.81). For all matches, similar patterns were seen across subgroups. No effect of DrotAA was seen for two organs failing or lower severity scores, compared with a significant mortality reduction for three or more organs failing or higher severity scores. CONCLUSION: Use of DrotAA was approximately one in 16 for admissions meeting the definition for severe sepsis and with two or more organs failing. Patients receiving DrotAA were younger and more severely ill but were less likely to have serious conditions in their past medical history. Nonrandomized estimates for the effectiveness of DrotAA were consistent with the findings of PROWESS. DrotAA appeared not to be effective in patients with less severe disease. PMID- 18430216 TI - Distribution of transferrin binding protein B gene (tbpB) variants among Neisseria species. AB - BACKGROUND: Transferrin binding protein B (tbpB), an outer membrane lipoprotein, is required for the acquisition of iron from human transferrin. Two tbpB families have been documented in Neisseria meningitidis: an isotype I tbpB gene of 1.8 kb and an isotype II tbpB gene of 2.1 kb, the former expressed by meningococci in the disease-associated ST-11 clonal complex and the latter found among meningococci belonging to the hyper-invasive clonal complexes including ST-8, ST 18, ST-32, ST-41/44 as well as N. gonorrhoeae isolates. The origin of the isotype I tbpB gene is unknown, however several features in common with non-pathogenic Neisseria and the ST-11 clonal complex N. meningitidis isolate FAM18 have been documented leading to the hypothesis that the isotype I tbpB gene may also be shared between non-pathogenic Neisseria and ST-11 meningococci. As a result, the diversity of the tbpB gene was investigated in a defined collection of Neisseria species. RESULTS: Two families of isotype I tbpB were identified: family A containing conserved genes belonging to ST-11 meningococci, N. polysaccharea and N. lactamica isolates and family B including more diverse isotype I tbpB genes from N. sicca, N. mucosa, N. flava, N. subflava as well as N. cinerea, N. flavescens and N. polysaccharea isolates. Three isotype II tbpB families were identified with: family C containing diverse tbpB genes belonging to N. polysaccharea, N. lactamica, N. gonorrhoeae and N. meningitidis isolates, family D including another subset of isotype II tbpB genes from N. lactamica isolates and family E solely composed of N. gonorrhoeae tbpB genes. CONCLUSION: This study reveals another instance of similarity between meningococci of the ST-11 clonal complex and non-pathogenic Neisseria with the origin of the isotype I tbpB gene resulting from a horizontal genetic transfer event occurring between these two populations. PMID- 18430217 TI - Bracing patients with idiopathic scoliosis: design of the Dutch randomized controlled treatment trial. AB - BACKGROUND: The effectiveness of bracing patients with IS has not yet been convincingly established due to a lack of RCTs. Some authors suggest that their results confirm that bracing is effective; others conclude that the effectiveness of bracing is doubtful or recommend a RCT. The aim of this study was to establish whether bracing patients with idiopathic scoliosis (IS) in an early stage will result in at least 5 degrees less mean progression of the curvature compared to the control group after two years of follow-up. METHODS: A randomized controlled trial was designed. Eligible patients are girls and boys in the age group 8-15 years whose diagnosis of IS has been established by an orthopedic surgeon, who have not yet been treated by bracing or surgery, and for whom further growth of physical height is still expected based on medical examination and maturation characteristics (Risser < or = 2). The Cobb angle of the eligible patient should either be minimally 22 and maximally 29 degrees with established progression of more than 5 degrees, or should be minimally 30 and maximally 35 degrees; established progression for the latter is not necessary. A total of 100 patients will be included in this trial. The intervention group will be treated with full time Boston brace wear; the control group will not be braced. Every four months, each patient will have a physical and an X-ray examination. The main outcomes will be the Cobb angle two years after inclusion and health-related quality of life. DISCUSSION: The results of this trial will be of great importance for the discussion on early treatment for scoliosis. Furthermore, the result will also be important for screening for scoliosis policies. TRIAL REGISTRATION: Nederlands Trialregister ISRCTN36964733. PMID- 18430218 TI - Promoting prevention with economic arguments - the case of Finnish occupational health services. AB - BACKGROUND: Both social and ethical arguments have been used to support preventive occupational health services (OHS). During the 1990s it became more common to support political argumentation for occupational health and safety by converting the consequences of ill health at work into monetary units. In addition, OHS has been promoted as a profitable investment for companies, and this aspect has been used by OHS providers in their marketing. Our intention was to study whether preventive occupational health services positively influence a company's economic performance. METHODS: We combined the financial statements provided by Statistics Finland and employers' reimbursement applications for occupational health services (OHS) costs to the Social Insurance Institution. The data covered the years 1997, 1999 and 2001 and over 6000 companies. We applied linear regression analysis to assess whether preventive OHS had a positive influence on the companies' economic performance after two or four years. RESULTS: Resources invested in preventive OHS were not positively related to a company's economic performance. In fact, the total cost of preventive OHS per turnover was negatively correlated to economic performance. CONCLUSION: Even if OHS has no effect on the economic performance of companies, it may have other effects more specific to OHS. Therefore, we recommend that the evaluation of prevention in OHS should move towards outcome measures, such as sickness absence, disability pension and productivity, when applicable, both in occupational health service research and in practice at workplaces. PMID- 18430219 TI - Incorporating health care quality into health antitrust law. AB - BACKGROUND: Antitrust authorities treat price as a proxy for hospital quality since health care quality is difficult to observe. As the ability to measure quality improved, more research became necessary to investigate the relationship between hospital market power and patient outcomes. This paper examines the impact of hospital competition on the quality of care as measured by the risk adjusted mortality rates with the hospital as the unit of analysis. The study separately examines the effect of competition on non-profit hospitals. METHODS: We use California Office of Statewide Health Planning and Development (OSHPD) data from 1997 through 2002. Empirical model is a cross-sectional study of 373 hospitals. Regression analysis is used to estimate the relationship between Coronary Artery Bypass Graft (CABG) risk-adjusted mortality rates and hospital competition. RESULTS: Regression results show lower risk-adjusted mortality rates in the presence of a more competitive environment. This result holds for all alternative hospital market definitions. Non-profit hospitals do not have better patient outcomes than investor-owned hospitals. However, they tend to provide better quality in less competitive environments. CABG volume did not have a significant effect on patient outcomes. CONCLUSION: Quality should be incorporated into the antitrust analysis. When mergers lead to higher prices and lower quality, thus lower social welfare, the antitrust challenge of hospital mergers is warranted. The impact of lower hospital competition on quality of care delivered by non-profit hospitals is ambiguous. PMID- 18430220 TI - Application of reinforcement learning for segmentation of transrectal ultrasound images. AB - BACKGROUND: Among different medical image modalities, ultrasound imaging has a very widespread clinical use. But, due to some factors, such as poor image contrast, noise and missing or diffuse boundaries, the ultrasound images are inherently difficult to segment. An important application is estimation of the location and volume of the prostate in transrectal ultrasound (TRUS) images. For this purpose, manual segmentation is a tedious and time consuming procedure. METHODS: We introduce a new method for the segmentation of the prostate in transrectal ultrasound images, using a reinforcement learning scheme. This algorithm is used to find the appropriate local values for sub-images and to extract the prostate. It contains an offline stage, where the reinforcement learning agent uses some images and manually segmented versions of these images to learn from. The reinforcement agent is provided with reward/punishment, determined objectively to explore/exploit the solution space. After this stage, the agent has acquired knowledge stored in the Q-matrix. The agent can then use this knowledge for new input images to extract a coarse version of the prostate. RESULTS: We have carried out experiments to segment TRUS images. The results demonstrate the potential of this approach in the field of medical image segmentation. CONCLUSION: By using the proposed method, we can find the appropriate local values and segment the prostate. This approach can be used for segmentation tasks containing one object of interest. To improve this prototype, more investigations are needed. PMID- 18430221 TI - Gene expression profiling in primary breast cancer distinguishes patients developing local recurrence after breast-conservation surgery, with or without postoperative radiotherapy. AB - INTRODUCTION: Some patients with breast cancer develop local recurrence after breast-conservation surgery despite postoperative radiotherapy, whereas others remain free of local recurrence even in the absence of radiotherapy. As clinical parameters are insufficient for identifying these two groups of patients, we investigated whether gene expression profiling would add further information. METHODS: We performed gene expression analysis (oligonucleotide arrays, 26,824 reporters) on 143 patients with lymph node-negative disease and tumor-free margins. A support vector machine was employed to build classifiers using leave one-out cross-validation. RESULTS: Within the estrogen receptor-positive (ER+) subgroup, the gene expression profile clearly distinguished patients with local recurrence after radiotherapy (n = 20) from those without local recurrence (n = 80 with or without radiotherapy). The receiver operating characteristic (ROC) area was 0.91, and 5,237 of 26,824 reporters had a P value of less than 0.001 (false discovery rate = 0.005). This gene expression profile provides substantially added value to conventional clinical markers (for example, age, histological grade, and tumor size) in predicting local recurrence despite radiotherapy. Within the ER- subgroup, a weaker, but still significant, signal was found (ROC area = 0.74). The ROC area for distinguishing patients who develop local recurrence from those who remain local recurrence-free in the absence of radiotherapy was 0.66 (combined ER+/ER-). CONCLUSION: A highly distinct gene expression profile for patients developing local recurrence after breast conservation surgery despite radiotherapy has been identified. If verified in further studies, this profile might be a most important tool in the decision making for surgery and adjuvant therapy. PMID- 18430222 TI - GAPscreener: an automatic tool for screening human genetic association literature in PubMed using the support vector machine technique. AB - BACKGROUND: Synthesis of data from published human genetic association studies is a critical step in the translation of human genome discoveries into health applications. Although genetic association studies account for a substantial proportion of the abstracts in PubMed, identifying them with standard queries is not always accurate or efficient. Further automating the literature-screening process can reduce the burden of a labor-intensive and time-consuming traditional literature search. The Support Vector Machine (SVM), a well-established machine learning technique, has been successful in classifying text, including biomedical literature. The GAPscreener, a free SVM-based software tool, can be used to assist in screening PubMed abstracts for human genetic association studies. RESULTS: The data source for this research was the HuGE Navigator, formerly known as the HuGE Pub Lit database. Weighted SVM feature selection based on a keyword list obtained by the two-way z score method demonstrated the best screening performance, achieving 97.5% recall, 98.3% specificity and 31.9% precision in performance testing. Compared with the traditional screening process based on a complex PubMed query, the SVM tool reduced by about 90% the number of abstracts requiring individual review by the database curator. The tool also ascertained 47 articles that were missed by the traditional literature screening process during the 4-week test period. We examined the literature on genetic associations with preterm birth as an example. Compared with the traditional, manual process, the GAPscreener both reduced effort and improved accuracy. CONCLUSION: GAPscreener is the first free SVM-based application available for screening the human genetic association literature in PubMed with high recall and specificity. The user friendly graphical user interface makes this a practical, stand-alone application. The software can be downloaded at no charge. PMID- 18430223 TI - The origin and evolution of the ribosome. AB - BACKGROUND: The origin and early evolution of the active site of the ribosome can be elucidated through an analysis of the ribosomal proteins' taxonomic block structures and their RNA interactions. Comparison between the two subunits, exploiting the detailed three-dimensional structures of the bacterial and archaeal ribosomes, is especially informative. RESULTS: The analysis of the differences between these two sites can be summarized as follows: 1) There is no self-folding RNA segment that defines the decoding site of the small subunit; 2) there is one self-folding RNA segment encompassing the entire peptidyl transfer center of the large subunit; 3) the protein contacts with the decoding site are made by a set of universal alignable sequence blocks of the ribosomal proteins; 4) the majority of those peptides contacting the peptidyl transfer center are made by bacterial or archaeal-specific sequence blocks. CONCLUSION: These clear distinctions between the two subunit active sites support an earlier origin for the large subunit's peptidyl transferase center (PTC) with the decoding site of the small subunit being a later addition to the ribosome. The main implications are that a single self-folding RNA, in conjunction with a few short stabilizing peptides, formed the precursor of the modern ribosomal large subunit in association with a membrane. PMID- 18430224 TI - Unexpected complexity of the aquaporin gene family in the moss Physcomitrella patens. AB - BACKGROUND: Aquaporins, also called major intrinsic proteins (MIPs), constitute an ancient superfamily of channel proteins that facilitate the transport of water and small solutes across cell membranes. MIPs are found in almost all living organisms and are particularly abundant in plants where they form a divergent group of proteins able to transport a wide selection of substrates. RESULTS: Analyses of the whole genome of Physcomitrella patens resulted in the identification of 23 MIPs, belonging to seven different subfamilies, of which only five have been previously described. Of the newly discovered subfamilies one was only identified in P. patens (Hybrid Intrinsic Protein, HIP) whereas the other was found to be present in a wide variety of dicotyledonous plants and forms a major previously unrecognized MIP subfamily (X Intrinsic Proteins, XIPs). Surprisingly also some specific groups within subfamilies present in Arabidopsis thaliana and Zea mays could be identified in P. patens. CONCLUSION: Our results suggest an early diversification of MIPs resulting in a large number of subfamilies already in primitive terrestrial plants. During the evolution of higher plants some of these subfamilies were subsequently lost while the remaining subfamilies expanded and in some cases diversified, resulting in the formation of more specialized groups within these subfamilies. PMID- 18430225 TI - Widespread duplications in the genomes of laboratory stocks of Dictyostelium discoideum. AB - BACKGROUND: Duplications of stretches of the genome are an important source of individual genetic variation, but their unrecognized presence in laboratory organisms would be a confounding variable for genetic analysis. RESULTS: We report here that duplications of 15 kb or more are common in the genome of the social amoeba Dictyostelium discoideum. Most stocks of the axenic 'workhorse' strains Ax2 and Ax3/4 obtained from different laboratories can be expected to carry different duplications. The auxotrophic strains DH1 and JH10 also bear previously unreported duplications. Strain Ax3/4 is known to carry a large duplication on chromosome 2 and this structure shows evidence of continuing instability; we find a further variable duplication on chromosome 5. These duplications are lacking in Ax2, which has instead a small duplication on chromosome 1. Stocks of the type isolate NC4 are similarly variable, though we have identified some approximating the assumed ancestral genotype. More recent wild-type isolates are almost without large duplications, but we can identify small deletions or regions of high divergence, possibly reflecting responses to local selective pressures. Duplications are scattered through most of the genome, and can be stable enough to reconstruct genealogies spanning decades of the history of the NC4 lineage. The expression level of many duplicated genes is increased with dosage, but for others it appears that some form of dosage compensation occurs. CONCLUSION: The genetic variation described here must underlie some of the phenotypic variation observed between strains from different laboratories. We suggest courses of action to alleviate the problem. PMID- 18430226 TI - Interaction of circadian clock proteins PER2 and CRY with BMAL1 and CLOCK. AB - BACKGROUND: Circadian oscillation of clock-controlled gene expression is mainly regulated at the transcriptional level. Heterodimers of CLOCK and BMAL1 act as activators of target gene transcription; however, interactions of PER and CRY proteins with the heterodimer abolish its transcriptional activation capacity. PER and CRY are therefore referred to as negative regulators of the circadian clock. To further elucidate the mechanism how positive and negative components of the clock interplay, we characterized the interactions of PER2, CRY1 and CRY2 with BMAL1 and CLOCK using a mammalian two-hybrid system and co immunoprecipitation assays. RESULTS: Both PER2 and the CRY proteins were found to interact with BMAL1 whereas only PER2 interacts with CLOCK. CRY proteins seem to have a higher affinity to BMAL1 than PER2. Moreover, we provide evidence that PER2, CRY1 and CRY2 bind to different domains in the BMAL1 protein. CONCLUSION: The regulators of clock-controlled transcription PER2, CRY1 and CRY2 differ in their capacity to interact with each single component of the BMAL1-CLOCK heterodimer and, in the case of BMAL1, also in their interaction sites. Our data supports the hypothesis that CRY proteins, especially CRY1, are stronger repressors than PER proteins. PMID- 18430227 TI - Framingham Stroke Risk Profile and poor cognitive function: a population-based study. AB - BACKGROUND: The relationship between stroke risk and cognitive function has not previously been examined in a large community living sample other than the Framingham cohort. The objective of this study was to examine the relationship between 10-year risk for incident stroke and cognitive function in a large population-based sample. METHODS: Participants were 7377 adults aged 50 years and over of the 2002 wave of the English Longitudinal Study of Ageing, a prospective cohort study. A modified version of the Framingham Stroke Risk Profile (incorporating age, sex, systolic blood pressure, antihypertensive medication, diabetes, smoking status, cardiovascular disease, and atrial fibrillation) was used to assess 10-year risk of stroke. Linear regression models were used to determine the cross-sectional relationship of stroke risk to global cognitive function and performance in multiple cognitive domains. RESULTS: In unadjusted models 10 percentage point increments of 10-year stroke risk were associated with poor global cognitive function (-0.40 SD units, 95% CI -0.43 - -0.38), and lowered performance in all cognitive domains. After statistical adjustment for age, sex, testing interval and other correlates of cognitive function the association with stroke risk was attenuated though remained significant for global cognitive function (-0.06 SD units, 95% CI -0.09 - -0.03), immediate and delayed verbal memory, semantic verbal fluency and processing speed. CONCLUSION: In individuals free from a history of stroke or dementia, high subclinical cerebrovascular disease burden was associated with worse cognitive function in multiple domains. PMID- 18430228 TI - Accidental carbon monoxide poisoning presenting without a history of exposure: a case report. AB - INTRODUCTION: Carbon monoxide poisoning is easy to diagnose when there is a history of exposure. When the exposure history is absent, or delayed, the diagnosis is more difficult and relies on recognising the importance of multi system disease. We present a case of accidental carbon monoxide poisoning. CASE PRESENTATION: A middle-aged man, who lived alone in his mobile home was found by friends in a confused, incontinent state. Initial signs included respiratory failure, cardiac ischaemia, hypotension, encephalopathy and a rash, whilst subsequent features included rhabdomyolysis, renal failure, amnesia, dysarthria, parkinsonism, peripheral neuropathy, supranuclear gaze palsy and cerebral haemorrhage. Despite numerous investigations including magnetic resonance cerebral imaging, lumbar puncture, skin biopsy, muscle biopsy and electroencephalogram a diagnosis remained elusive. Several weeks after admission, diagnostic breakthrough was achieved when the gradual resolution of the patient's amnesia, encephalopathy and dysarthria allowed an accurate history to be taken for the first time. The patient's last recollection was turning on his gas heating for the first time since the spring. A gas heating engineer found the patient's gas boiler to be in a dangerous state of disrepair and it was immediately decommissioned. CONCLUSION: This case highlights several important issues: the bewildering myriad of clinical features of carbon monoxide poisoning, the importance of making the diagnosis even at a late stage and preventing the patient's return to a potentially fatal toxic environment, and the paramount importance of the history in the diagnostic method. PMID- 18430229 TI - A systematic review of the clinical effectiveness of acupuncture for allergic rhinitis. AB - BACKGROUND: Allergies cause a considerable burden to both sufferers and the National Health Service. There is growing interest in acupuncture as a treatment for a range of conditions. Since acupuncture may modulate the immune system it could be a useful treatment for allergic rhinitis (AR) sufferers. We therefore assessed the evidence for the clinical effectiveness of acupuncture in patients with AR by performing a systematic review of the literature. METHODS: Searches (to 2007) were conducted in all major databases for randomised controlled trials (RCTs) evaluating the clinical effectiveness of acupuncture in the treatment of AR. No limits were placed on language. Studies were included if they compared acupuncture to a sham or inactive acupuncture treatment (placebo) with or without standard care. Meta-analysis was performed where feasible. RESULTS: Seven relevant RCTs were included after screening and application of inclusion and exclusion criteria. The trials were generally of poor quality as assessed by a modified Jadad scale, with the exception of two studies which scored highly. A wide variety of outcomes was measured but most assessed symptom severity on a visual analogue scale. A meta-analysis failed to show any summary benefits of acupuncture treatment for symptom severity scores or serum IgE measures which could not have been accounted for by chance alone. Acupuncture was not associated with any additional adverse events in the trials. CONCLUSION: There is currently insufficient evidence to support or refute the use of acupuncture in patients with AR. A large well conducted RCT, which overcomes identified methodological problems in the existing RCTs, would be required to resolve this question. PMID- 18430230 TI - Proliferating mesodermal cells in murine embryos exhibiting macrophage and lymphendothelial characteristics. AB - BACKGROUND: The data on the embryonic origin of lymphatic endothelial cells (LECs) from either deep embryonic veins or mesenchymal (or circulating) lymphangioblasts presently available remain inconsistent. In various vertebrates, markers for LECs are first expressed in specific segments of embryonic veins arguing for a venous origin of lymph vessels. Very recently, studies on the mouse have strongly supported this view. However, in the chick, we have observed a dual origin of LECs from veins and from mesodermal lymphangioblasts. Additionally, in murine embryos we have detected mesenchymal cells that co-express LEC markers and the pan-leukocyte marker CD45. Here, we have characterized the mesoderm of murine embryos with LEC markers Prox1, Lyve-1 and LA102 in combination with macrophage markers CD11b and F4/80. RESULTS: We observed cells co-expressing both types of markers (e.g. Prox1 - Lyve-1 - F4/80 triple-positive) located in the mesoderm, immediately adjacent to, and within lymph vessels. Our proliferation studies with Ki-67 antibodies showed high proliferative capacities of both the Lyve-1-positive LECs of lymph sacs/lymphatic sprouts and the Lyve-1-positive mesenchymal cells. CONCLUSION: Our data argue for a dual origin of LECs in the mouse, although the primary source of embryonic LECs may reside in specific embryonic veins and mesenchymal lymphangioblasts integrated secondarily into lymph vessels. The impact of a dual source of LECs for ontogenetic, phylogenetic and pathological lymphangiogenesis is discussed. PMID- 18430231 TI - A five-year retrospective study of the epidemiological characteristics and visual outcomes of patients hospitalized for ocular trauma in a Mediterranean area. AB - BACKGROUND: To determine the epidemiological characteristics and visual outcome of ocular trauma in southern Italy. METHODS: All cases of ocular trauma admitted to Department of Ophthalmology of Palermo University, Italy, from January 2001 December 2005 were retrospectively reviewed for open- or closed-globe injury (OGI or CGI). Data extracted included age, sex, residence, initial and final visual acuity (VA), cause and treatment of injury, hospitalization. The injuries were classified by Ocular Trauma Classification System (OTCS) and Birmingham Eye Trauma Terminology (BETT). We also referred to the Ocular Trauma Score (OTS) in evaluating the final visual outcome. RESULTS: Of the 298 eyes, there were 146 OGI and 152 CGI. Fifty eyes (16.8%) had an intraocular foreign body (IOFB). The annual incidence of eye injuries was 4.9 per 100,000. Most injuries occurred in men (84.6%, p < 0.0005), with an average age of 33.0 vs. 49.9 for women (p = 0.005). Cause of injury differed significantly by gender (p = 0.001) and urban vs. rural location (p = 0.009). The most frequent causes in men were outdoor activities related injuries (30.9%), work-related (25.4%), and sport-related (17.5%), and in women were home-related (52.2%) and outdoor activities related injuries (30.4%). In urban areas, road accidents were more frequent; in rural areas, work-related injuries were more frequent with a greater rate of IOFBs than in urban areas (p = 0.002). The incidence of OGI and CGI differed in work-related injuries (p < 0.0005), sport-related injuries (p < 0.0005), and assaults (p = 0.033). The final visual acuity was 20/40 (6/12) or better in 144 eyes (48.3%), 20/40-20/200 (6/12-6/60) in 90 eyes (30.2%), and <20/200 (6/60) or less in 46 eyes (15.5%). Eighteen eyes (6%) had a final acuity of no light perception. Of those eyes that presented with hand motion vision or better, 220 (86.6%) had a final vision of better than 20/200 (6/60). Initial visual acuity was found to be correlated with final visual acuity (Spearman's correlation coefficient = 0.658; p < 0.001). The likelihood of the final visual acuities in the OTS categories was correlated to that of the OTS study group in 12 of 14 cases (85.7%). CONCLUSION: This analysis provides insight into the epidemiology of patients hospitalized for ocular trauma. The findings indicate that ocular trauma is a significant cause of visual loss in this population. PMID- 18430233 TI - Pregnancy following homologous prepubertal ovarian transplantation in the dog. AB - In several canine models of hereditary human disease the homozygote dogs die prior to puberty, or have substantially reduced fertility. To create a clinically healthy animal that can be bred, but can also transmit the gene of interest, a model of homologous ovarian transplantation in prepubertal dogs was developed. Six dog leukocyte antigen (DLA) identical littermates underwent transplantation of ovarian cortical strips (n = 2) or the entire ovary (n = 4). Immunosuppression was maintained with cyclosporine and MMF in the immediate post-operative period and cyclosporine alone thereafter. All 6 dogs entered puberty and normal semiannual estrus cycles as demonstrated by both physical changes and increasing serum progesterone. Four dogs were bred to a proven stud male, and one became pregnant. Three viable fetuses with observable heart-beats were detected on ultrasound examination. Although the dog eventually aborted the litter, this work represents the first pregnancy achieved following a prepubertal ovarian transplant in the dog. PMID- 18430232 TI - Roots of angiosperm formins: the evolutionary history of plant FH2 domain containing proteins. AB - BACKGROUND: Shuffling of modular protein domains is an important source of evolutionary innovation. Formins are a family of actin-organizing proteins that share a conserved FH2 domain but their overall domain architecture differs dramatically between opisthokonts (metazoans and fungi) and plants. We performed a phylogenomic analysis of formins in most eukaryotic kingdoms, aiming to reconstruct an evolutionary scenario that may have produced the current diversity of domain combinations with focus on the origin of the angiosperm formin architectures. RESULTS: The Rho GTPase-binding domain (GBD/FH3) reported from opisthokont and Dictyostelium formins was found in all lineages except plants, suggesting its ancestral character. Instead, mosses and vascular plants possess the two formin classes known from angiosperms: membrane-anchored Class I formins and Class II formins carrying a PTEN-like domain. PTEN-related domains were found also in stramenopile formins, where they have been probably acquired independently rather than by horizontal transfer, following a burst of domain rearrangements in the chromalveolate lineage. A novel RhoGAP-related domain was identified in some algal, moss and lycophyte (but not angiosperm) formins that define a specific branch (Class III) of the formin family. CONCLUSION: We propose a scenario where formins underwent multiple domain rearrangements in several eukaryotic lineages, especially plants and chromalveolates. In plants this replaced GBD/FH3 by a probably inactive RhoGAP-like domain, preserving a formin mediated association between (membrane-anchored) Rho GTPases and the actin cytoskeleton. Subsequent amplification of formin genes, possibly coincident with the expansion of plants to dry land, was followed by acquisition of alternative membrane attachment mechanisms present in extant Class I and Class II formins, allowing later loss of the RhoGAP-like domain-containing formins in angiosperms. PMID- 18430234 TI - Postnatal stem/progenitor cells derived from the dental pulp of adult chimpanzee. AB - BACKGROUND: Chimpanzee dental pulp stem/stromal cells (ChDPSCs) are very similar to human bone marrow derived mesenchymal stem/stromal cells (hBMSCs) as demonstrated by the expression pattern of cell surface markers and their multipotent differentiation capability. RESULTS: ChDPSCs were isolated from an incisor and a canine of a forty-seven year old female chimpanzee. A homogenous population of ChDPSCs was established in early culture at a high proliferation rate and verified by the expression pattern of thirteen cell surface markers. The ChDPSCs are multipotent and were capable of differentiating into osteogenic, adipogenic and chondrogenic lineages under appropriate in vitro culture conditions. ChDPSCs also express stem cell (Sox-2, Nanog, Rex-1, Oct-4) and osteogenic (Osteonectin, osteocalcin, osteopontin) markers, which is comparable to reported results of rhesus monkey BMSCs (rBMSCs), hBMSCs and hDPSCs. Although ChDPSCs vigorously proliferated during the initial phase and gradually decreased in subsequent passages, the telomere length indicated that telomerase activity was not significantly reduced. CONCLUSION: These results demonstrate that ChDPSCs can be efficiently isolated from post-mortem teeth of adult chimpanzees and are multipotent. Due to the almost identical genome composition of humans and chimpanzees, there is an emergent need for defining the new role of chimpanzee modeling in comparative medicine. Teeth are easy to recover at necropsy and easy to preserve prior to the retrieval of dental pulp for stem/stromal cells isolation. Therefore, the establishment of ChDPSCs would preserve and maximize the applications of such a unique and invaluable animal model, and could advance the understanding of cellular functions and differentiation control of adult stem cells in higher primates. PMID- 18430235 TI - The many faces of ubiquitinated histone H2A: insights from the DUBs. AB - Monoubiquitination of H2A is a major histone modification in mammalian cells. Understanding how monoubiquitinated H2A (uH2A) regulates DNA-based processes in the context of chromatin is a challenging question. Work in the past years linked uH2A to transcriptional repression by the Polycomb group proteins of developmental regulators. Recently, a number of mammalian deubiquitinating enzymes (DUBs) that catalyze the removal of ubiquitin from H2A have been discovered. These studies provide convincing evidence that H2A deubiquitination is connected with gene activation. In addition, uH2A regulatory enzymes have crucial roles in the cellular response to DNA damage and in cell cycle progression. In this review we will discuss new insights into uH2A biology, with emphasis on the H2A DUBs. PMID- 18430237 TI - Assessment of an undergraduate psychiatry course in an African setting. AB - BACKGROUND: International reports recommend the improvement in the amount and quality of training for mental health workers in low and middle income countries. The Scotland-Malawi Mental Health Education Project (SMMHEP) has been established to support the teaching of psychiatry to medical students in the University of Malawi. While anecdotally supportive medical educational initiatives appear of value, little quantitative evidence exists to demonstrate whether such initiatives can deliver comparable educational standards. This study aimed to assess the effectiveness of an undergraduate psychiatry course given by UK psychiatrists in Malawi by studying University of Malawi and Edinburgh University medical students' performance on an MCQ examination paper. METHODS: An undergraduate psychiatry course followed by an MCQ exam was delivered by the SMMHEP to 57 Malawi medical students. This same MCQ exam was given to 71 Edinburgh University medical students who subsequently sat their own Edinburgh University examination. RESULTS: There were no significant differences between Edinburgh students' performance on the Malawi exam and their own Edinburgh University exam. (p = 0.65). This would suggest that the Malawi exam is a comparable standard to the Edinburgh exam. Malawi students marks ranged from 52.4%-84.6%. Importantly 84.4% of Malawi students scored above 60% on their exam which would equate to a hypothetical pass by UK university standards. CONCLUSION: The support of an undergraduate course in an African setting by high income country specialists can attain a high percentage pass rate by UK standards. Although didactic teaching has been surpassed by more novel educational methods, in resource poor countries it remains an effective and cost effective method of gaining an important educational standard. PMID- 18430236 TI - Genetic and functional association of FAM5C with myocardial infarction. AB - BACKGROUND: We previously identified a 40 Mb region of linkage on chromosome 1q in our early onset coronary artery disease (CAD) genome-wide linkage scan (GENECARD) with modest evidence for linkage (n = 420, LOD 0.95). When the data are stratified by acute coronary syndrome (ACS), this modest maximum in the overall group became a well-defined LOD peak (maximum LOD of 2.17, D1S1589/D1S518). This peak overlaps a recently identified inflammatory biomarker (MCP-1) linkage region from the Framingham Heart Study (maximum LOD of 4.27, D1S1589) and a region of linkage to metabolic syndrome from the IRAS study (maximum LOD of 2.59, D1S1589/D1S518). The overlap of genetic screens in independent data sets provides evidence for the existence of a gene or genes for CAD in this region. METHODS: A peak-wide association screen (457 SNPs) was conducted of a region 1 LOD score down from the peak marker (168-198 Mb) in a linkage peak for acute coronary syndrome (ACS) on chromosome 1, within a family based early onset coronary artery disease (CAD) sample (GENECARD). RESULTS: Polymorphisms were identified within the 'family with sequence similarity 5, member C' gene (FAM5C) that show genetic linkage to and are associated with myocardial infarction (MI) in GENECARD. The association was confirmed in an independent CAD case-control sample (CATHGEN) and strong association with MI was identified with single nucleotide polymorphisms (SNPs) in the 3' end of FAM5C. FAM5C genotypes were also correlated with expression of the gene in human aorta. Expression levels of FAM5C decreased with increasing passage of proliferating aortic smooth muscle cells (SMC) suggesting a role for this molecule in smooth muscle cell proliferation and senescence. CONCLUSION: These data implicate FAM5C alleles in the risk of myocardial infarction and suggest further functional studies of FAM5C are required to identify the gene's contribution to atherosclerosis. PMID- 18430238 TI - Differential protein expression following low temperature culture of suspension CHO-K1 cells. AB - BACKGROUND: To ensure maximal productivity of recombinant proteins (rP) during production culture it is typical to encourage an initial phase of rapid cell proliferation to achieve high biomass followed by a stationary phase where cellular energies are directed towards production of rP. During many such biphasic cultures, the initial phase of rapid cell growth at 37 degrees C is followed by a growth arrest phase induced through reduction of the culture temperature. Low temperature induced growth arrest is associated with many positive phenotypes including increased productivity, sustained viability and an extended production phase, although the mechanisms regulating these phenotypes during mild hypothermia are poorly understood. RESULTS: In this study differential protein expression in suspension CHO-K1 cells was investigated following a reduction of the culture temperature from 37 degrees C to 31 degrees C in comparison to standard batch culture maintained at 37 degrees C using 2D DIGE (Fluorescence 2-D Difference Gel Electrophoresis) and mass spectrometry (MS). There is only limited proteomic analysis of suspension-grown CHO cells describing a direct comparison of temperature shifted versus non-temperature shifted cultures using 2D-DIGE. This investigation has enabled the identification of temperature-dependent as well as temperature-independent proteomic changes. 201 proteins were observed as differentially expressed following temperature shift, of which 118 were up regulated. Of the 53 proteins identified by MALDI-ToF MS, 23 were specifically differentially expressed upon reduction of the culture temperature and were found related to a variety of cellular functions such as regulation of growth (HNRPC), cap-independent translation (EIF4A), apoptosis (importin-alpha), the cytoskeleton (vimentin) and glycoprotein quality control (alpha glucosidase 2). CONCLUSION: These results indicate the extent of the temperature response in CHO-K1 cells and suggest a number of key regulatory proteins and pathways that are involved in modulating the response of cells to mild hypothermia. Regulation of these identified proteins and pathways could be useful for future approaches to engineer CHO cells for improved recombinant protein production. PMID- 18430239 TI - Equity and health policy in Africa: using concept mapping in Moore (Burkina Faso). AB - BACKGROUND: This methodological article is based on a health policy research project conducted in Burkina Faso (West Africa). Concept mapping (CM) was used as a research method to understand the local views of equity among stakeholders, who were concerned by the health policy under consideration. While this technique has been used in North America and elsewhere, to our knowledge it has not yet been applied in Africa in any vernacular language. Its application raises many issues and certain methodological limitations. Our objective in this article is to present its use in this particular context, and to share a number of methodological observations on the subject. METHODS: Two CMs were done among two different groups of local stakeholders following four steps: generating ideas, structuring the ideas, computing maps using multidimensional scaling and cluster analysis methods, and interpreting maps. Fifteen nurses were invited to take part in the study, all of whom had undergone training on health policies. Of these, nine nurses (60%) ultimately attended the two-day meeting, conducted in French. Of 45 members of village health committees who attended training on health policies, only eight were literate in the local language (Moore). Seven of these (88%) came to the meeting. RESULTS: The local perception of equity seems close to the egalitarian model. The actors are not ready to compromise social stability and peace for the benefit of the worst-off. The discussion on the methodological limitations of CM raises the limitations of asking a single question in Moore and the challenge of translating a concept as complex as equity. While the translation of equity into Moore undoubtedly oriented the discussions toward social relations, we believe that, in the context of this study, the open-ended question concerning social justice has a threefold relevance. At the same time, those limitations were transformed into strengths. We understand that it was essential to resort to the focus group approach to explore deeply a complex subject such as equity, which became, after the two CMs, one of the important topics of the research. CONCLUSION: Using this technique in a new context was not the easiest thing to do. Nevertheless, contrary to what local organizers thought when we explained to them this "crazy" idea of applying the technique in Moore with peasants, we believe we have shown that it was feasible, even with persons not literate in French. PMID- 18430240 TI - Chemical and physical properties of some saline lakes in Alberta and Saskatchewan. AB - BACKGROUND: The Northern Great Plains of Canada are home to numerous permanent and ephemeral athalassohaline lakes. These lakes display a wide range of ion compositions, salinities, stratification patterns, and ecosystems. Many of these lakes are ecologically and economically significant to the Great Plains Region. A survey of the physical characteristics and chemistry of 19 lakes was carried out to assess their suitability for testing new tools for determining past salinity from the sediment record. RESULTS: Data on total dissolved solids (TDS), specific conductivity, temperature, dissolved oxygen (DO), and pH were measured in June, 2007. A comparison of these data with past measurements indicates that salinity is declining at Little Manitou and Big Quill Lakes in the province of Saskatchewan. However salinity is rising at other lakes in the region, including Redberry and Manito Lakes. CONCLUSION: The wide range of salinities found across a small geographic area makes the Canadian saline lakes region ideal for testing salinity proxies. A nonlinear increase in salinity at Redberry Lake is likely influenced by its morphometry. This acceleration has ecological implications for the migratory bird species found within the Redberry Important Bird Area. PMID- 18430241 TI - Requirements for effective academic leadership in Iran: a nominal group technique exercise. AB - BACKGROUND: During the last two decades, medical education in Iran has shifted from elite to mass education, with a considerable increase in number of schools, faculties, and programs. Because of this transformation, it is a good case now to explore academic leadership in a non-western country. The objective of this study was to explore the views on effective academic leadership requirements held by key informants in Iran's medical education system. METHODS: A nominal group study was conducted by strategic sampling in which participants were requested to discuss and report on requirements for academic leadership, suggestions and barriers. Written notes from the discussions were transcribed and subjected to content analysis. RESULTS: Six themes of effective academic leadership emerged: 1)shared vision, goal, and strategy, 2) teaching and research leadership, 3) fair and efficient management, 4) mutual trust and respect, 5) development and recognition, and 6) transformational leadership. Current Iranian academic leadership suffers from lack of meritocracy, conservative leaders, politicization, bureaucracy, and belief in misconceptions. CONCLUSION: The structure of the Iranian medical university system is not supportive of effective academic leadership. However, participants' views on effective academic leadership are in line with what is also found in the western literature, that is, if the managers could create the premises for a supportive and transformational leadership, they could generate mutual trust and respect in academia and increase scientific production. PMID- 18430242 TI - A randomised clinical trial of subgrouping and targeted treatment for low back pain compared with best current care. The STarT Back Trial Study Protocol. AB - BACKGROUND: Back pain is a major health problem and many sufferers develop persistent symptoms. Detecting relevant subgroups of patients with non-specific low back pain has been highlighted as a priority area for research, as this could enable better secondary prevention through the targeting of prognostic indicators for persistent, disabling symptoms. We plan to conduct a randomised controlled trial to establish whether subgrouping using a novel tool, combined with targeted treatment, is better than best current care at reducing long-term disability from low back pain. METHODS/DESIGN: We will recruit 800 participants aged 18 years and over with non-specific low back pain from 8-10 GP practices within two Primary Care Trusts in Staffordshire, England. Our primary outcome measures are low back pain disability and catastrophising. Secondary outcomes include back pain intensity, global change, leg pain, fear avoidance, anxiety, depression, illness perceptions, patient satisfaction, overall health status and cost-effectiveness. Data will be collected before randomisation, and 4 and 12 months later. Participants are randomised to receive either newly developed interventions, delivered by trained physiotherapists and targeted according to subgroups defined by tool scores, or best current care. DISCUSSION: This paper presents detail on the rationale, design, methods and operational aspects of the trial. TRIAL REGISTRATION: Current Controlled Trials ISRCTN37113406. PMID- 18430243 TI - Disruption of four kinesin genes in dictyostelium. AB - BACKGROUND: Kinesin and dynein are the two families of microtubule-based motors that drive much of the intracellular movements in eukaryotic cells. Using a gene knockout strategy, we address here the individual function(s) of four of the 13 kinesin proteins in Dictyostelium. The goal of our ongoing project is to establish a minimal motility proteome for this basal eukaryote, enabling us to contrast motor functions here with the often far more elaborate motor families in the metazoans. RESULTS: We performed individual disruptions of the kinesin genes, kif4, kif8, kif10, and kif11. None of the motors encoded by these genes are essential for development or viability of Dictyostelium. Removal of Kif4 (kinesin 7; CENP-E family) significantly impairs the rate of cell growth and, when combined with a previously characterized dynein inhibition, results in dramatic defects in mitotic spindle assembly. Kif8 (kinesin-4; chromokinesin family) and Kif10 (kinesin-8; Kip3 family) appear to cooperate with dynein to organize the interphase radial microtubule array. CONCLUSION: The results reported here extend the number of kinesin gene disruptions in Dictyostelium, to now total 10, among the 13 isoforms. None of these motors, individually, are required for short-term viability. In contrast, homologs of at least six of the 10 kinesins are considered essential in humans. Our work underscores the functional redundancy of motor isoforms in basal organisms while highlighting motor specificity in more complex metazoans. Since motor disruption in Dictyostelium can readily be combined with other motility insults and stresses, this organism offers an excellent system to investigate functional interactions among the kinesin motor family. PMID- 18430244 TI - Comparison of two dependent within subject coefficients of variation to evaluate the reproducibility of measurement devices. AB - BACKGROUND: The within-subject coefficient of variation and intra-class correlation coefficient are commonly used to assess the reliability or reproducibility of interval-scale measurements. Comparison of reproducibility or reliability of measurement devices or methods on the same set of subjects comes down to comparison of dependent reliability or reproducibility parameters. METHODS: In this paper, we develop several procedures for testing the equality of two dependent within-subject coefficients of variation computed from the same sample of subjects, which is, to the best of our knowledge, has not yet been dealt with in the statistical literature. The Wald test, the likelihood ratio, and the score tests are developed. A simple regression procedure based on results due to Pitman and Morgan is constructed. Furthermore we evaluate the statistical properties of these methods via extensive Monte Carlo simulations. The methodologies are illustrated on two data sets; the first are the microarray gene expressions measured by two plat- forms; the Affymetrix and the Amersham. Because microarray experiments produce expressions for a large number of genes, one would expect that the statistical tests to be asymptotically equivalent. To explore the behaviour of the tests in small or moderate sample sizes, we illustrated the methodologies on data from computer-aided tomographic scans of 50 patients. RESULTS: It is shown that the relatively simple Wald's test (WT) is as powerful as the likelihood ratio test (LRT) and that both have consistently greater power than the score test. The regression test holds its empirical levels, and in some occasions is as powerful as the WT and the LRT. CONCLUSION: A comparison between the reproducibility of two measuring instruments using the same set of subjects leads naturally to a comparison of two correlated indices. The presented methodology overcomes the difficulty noted by data analysts that dependence between datasets would confound any inferences one could make about the differences in measures of reliability and reproducibility. The statistical tests presented in this paper have good properties in terms of statistical power. PMID- 18430245 TI - Deep sequencing of chicken microRNAs. AB - BACKGROUND: The use of new, deep sequencing technologies has greatly accelerated microRNA discovery. We have applied this approach to the identification of chicken microRNAs and to the comparison of microRNAs in chicken embryo fibroblasts (CEF) infected with Marek's disease virus (MDV) to those present in uninfected CEF. RESULTS: We obtained 125,463 high quality reads that showed an exact match to the chicken genome. The majority of the reads corresponded to previously annotated chicken microRNAs; however, the sequences of many potential novel microsRNAs were obtained. A comparison of the reads obtained in MDV infected and uninfected CEF indicates that infection does not significantly perturb the expression profile of microRNAs. Frequently sequenced microRNAs include miR-221/222, which are thought to play a role in growth and proliferation. A number of microRNAs (e.g., let-7, miR-199a-1, 26a) are expressed at lower levels in MDV-induced tumors, highlighting the potential importance of this class of molecules in tumorigenesis. CONCLUSION: Deep sequencing technology is highly suited for small RNA discovery. This approach is independent of comparative sequence analysis, which has been the primary method used to identify chicken microRNAs. Our results have confirmed the expression of many microRNAs identified by sequence similarity and identified a pool of candidate novel microRNAs. PMID- 18430246 TI - Prediction and analysis of nucleosome exclusion regions in the human genome. AB - BACKGROUND: Nucleosomes are the basic structural units of eukaryotic chromatin, and they play a significant role in regulating gene expression. Specific DNA sequence patterns are known, from empirical and theoretical studies, to influence DNA bending and flexibility, and have been shown to exclude nucleosomes. A whole genome localization of these patterns, and their analysis, can add important insights on the gene regulation mechanisms that depend upon the structure of chromatin in and around a gene. RESULTS: A whole genome annotation for nucleosome exclusion regions (NXRegions) was carried out on the human genome. Nucleosome exclusion scores (NXScores) were calculated individually for each nucleotide, giving a measure of how likely a specific nucleotide and its immediate neighborhood would impair DNA bending and, consequently, exclude nucleosomes. The resulting annotations were correlated with 19055 gene expression profiles. We developed a new method based on Grubbs' outliers test for ranking genes based on their tissue specificity, and correlated this ranking with NXScores. The results show a strong correlation between tissue specificity of a gene and the propensity of its promoter to exclude nucleosomes (the promoter region was taken as -1500 to +500 bp from the RefSeq-annotated transcription start site). In addition, NXScores correlated well with gene density, gene expression levels, and DNaseI hypersensitive sites. CONCLUSION: We present, for the first time, a whole genome prediction of nucleosome exclusion regions for the human genome (the data are available for download from Additional Materials). Nucleosome exclusion patterns are correlated with various factors that regulate gene expression, which emphasizes the need to include chromatin structural parameters in experimental analysis of gene expression. PMID- 18430247 TI - A novel function for the presenilin family member spe-4: inhibition of spermatid activation in Caenorhabditis elegans. AB - BACKGROUND: Sperm cells must regulate the timing and location of activation to maximize the likelihood of fertilization. Sperm from most species, including the nematode Caenorhabditis elegans, activate upon encountering an external signal. Activation for C. elegans sperm occurs as spermatids undergo spermiogenesis, a profound cellular reorganization that produces a pseudopod. Spermiogenesis is initiated by an activation signal that is transduced through a series of gene products. It is now clear that an inhibitory pathway also operates in spermatids, preventing their premature progression to spermatozoa and resulting in fine-scale control over the timing of activation. Here, we describe the involvement of a newly assigned member of the inhibitory pathway: spe-4, a homolog of the human presenilin gene PS1. The spe-4(hc196) allele investigated here was isolated as a suppressor of sterility of mutations in the spermiogenesis signal transduction gene spe-27. RESULTS: Through mapping, complementation tests, DNA sequencing, and transformation rescue, we determined that allele hc196 is a mutation in the spe-4 gene. Our data show that spe-4(hc196) is a bypass suppressor that eliminates the need for the spermiogenesis signal transduction. On its own, spe-4(hc196) has a recessive, temperature sensitive spermatogenesis-defective phenotype, with mutants exhibiting (i) defective spermatocytes, (ii) defective spermatids, (iii) premature spermatid activation, and (iv) spermatozoa defective in fertilization, in addition to a small number of functional sperm which appear normal microscopically. CONCLUSION: A fraction of the sperm from spe-4(hc196) mutant males progress directly to functional spermatozoa without the need for an activation signal, suggesting that spe-4 plays a role in preventing spermatid activation. Another fraction of spermatozoa from spe-4(hc196) mutants are defective in fertilization. Therefore, prematurely activated spermatozoa may have several defects: we show that they may be defective in fertilization, and earlier work showed that they obstruct sperm transfer from males at mating. hc196 is a hypomorphic allele of spe-4, and its newly-discovered role inhibiting spermiogenesis may involve known proteolytic and/or calcium regulatory aspects of presenilin function, or it may involve yet-to-be discovered functions. PMID- 18430248 TI - Surgical treatment of a rare primary renal carcinoid tumor with liver metastasis. AB - BACKGROUND: Carcinoid tumors are characteristically low grade malignant neoplasms with neuroendocrine differentiation that arise in various body sites, most commonly the lung and gastrointestinal tract, but less frequently the kidneys, breasts, ovaries, testes, prostate and other locations. We report a case of a carcinoid of renal origin with synchronous single liver metastases on radiological studies. CASE PRESENTATION: A 45 year-old patient who presented with abdominal pain was found on CT scan to have lesions in the right ovary, right kidney, and left hepatic lobe. CA-125, CEA, and CA 19-9 were within normal limits, as were preoperative liver function tests and renal function. Biopsy of the liver mass demonstrated metastatic neuroendocrine tumor. At laparotomy, the patient underwent total abdominal hysterectomy with bilateral salpingo oophorectomy, radical right nephrectomy with lymphadenectomy, and left hepatectomy. Pathology evaluation reported a right ovarian borderline serous tumor, well-differentiated neuroendocrine carcinoma of the kidney (carcinoid) with 2 positive retroperitoneal lymph nodes, and a single liver metastasis. Immunohistochemistry revealed that this lesion was positive for synaptophysin and CD56, but negative for chromogranin as well as CD10, CD7, and CD20, consistent with a well-differentiated neuroendocrine tumor. She is doing well one year after her initial surgery, with no evidence of tumor recurrence. CONCLUSION: Early surgical intervention, together with careful surveillance and follow-up, can achieve successful long-term outcomes in patients with this rare malignancy. PMID- 18430250 TI - Genome classification by gene distribution: an overlapping subspace clustering approach. AB - BACKGROUND: Genomes of lower organisms have been observed with a large amount of horizontal gene transfers, which cause difficulties in their evolutionary study. Bacteriophage genomes are a typical example. One recent approach that addresses this problem is the unsupervised clustering of genomes based on gene order and genome position, which helps to reveal species relationships that may not be apparent from traditional phylogenetic methods. RESULTS: We propose the use of an overlapping subspace clustering algorithm for such genome classification problems. The advantage of subspace clustering over traditional clustering is that it can associate clusters with gene arrangement patterns, preserving genomic information in the clusters produced. Additionally, overlapping capability is desirable for the discovery of multiple conserved patterns within a single genome, such as those acquired from different species via horizontal gene transfers. The proposed method involves a novel strategy to vectorize genomes based on their gene distribution. A number of existing subspace clustering and biclustering algorithms were evaluated to identify the best framework upon which to develop our algorithm; we extended a generic subspace clustering algorithm called HARP to incorporate overlapping capability. The proposed algorithm was assessed and applied on bacteriophage genomes. The phage grouping results are consistent overall with the Phage Proteomic Tree and showed common genomic characteristics among the TP901-like, Sfi21-like and sk1-like phage groups. Among 441 phage genomes, we identified four significantly conserved distribution patterns structured by the terminase, portal, integrase, holin and lysin genes. We also observed a subgroup of Sfi21-like phages comprising a distinctive divergent genome organization and identified nine new phage members to the Sfi21 like genus: Staphylococcus 71, phiPVL108, Listeria A118, 2389, Lactobacillus phi AT3, A2, Clostridium phi3626, Geobacillus GBSV1, and Listeria monocytogenes PSA. CONCLUSION: The method described in this paper can assist evolutionary study through objectively classifying genomes based on their resemblance in gene order, gene content and gene positions. The method is suitable for application to genomes with high genetic exchange and various conserved gene arrangement, as demonstrated through our application on phages. PMID- 18430251 TI - VariVis: a visualisation toolkit for variation databases. AB - BACKGROUND: With the completion of the Human Genome Project and recent advancements in mutation detection technologies, the volume of data available on genetic variations has risen considerably. These data are stored in online variation databases and provide important clues to the cause of diseases and potential side effects or resistance to drugs. However, the data presentation techniques employed by most of these databases make them difficult to use and understand. RESULTS: Here we present a visualisation toolkit that can be employed by online variation databases to generate graphical models of gene sequence with corresponding variations and their consequences. The VariVis software package can run on any web server capable of executing Perl CGI scripts and can interface with numerous Database Management Systems and "flat-file" data files. VariVis produces two easily understandable graphical depictions of any gene sequence and matches these with variant data. While developed with the goal of improving the utility of human variation databases, the VariVis package can be used in any variation database to enhance utilisation of, and access to, critical information. PMID- 18430249 TI - Expression patterns and prognostic value of Bag-1 and Bcl-2 in breast cancer. AB - INTRODUCTION: Bcl-2 antanogene-1 (Bag-1) binds the anti-apoptotic mediator Bcl-2, and enhances its activity. Bcl-2 and Bag-1 are associated with chemotherapy resistance in cancer cells. Drugs that target Bcl-2 are currently in clinical development. The purpose of the present study was to examine expression patterns of Bag-1 in a large cohort of breast tumors and to assess the association with Bcl-2, estrogen receptor, progesterone receptor and Her2/neu, and other clinical/pathological variables. METHODS: Tissue microarrays containing primary specimens from 638 patients with 10-year follow-up were employed, and the expression of Bag-1, Bcl-2, estrogen receptor, progesterone receptor and Her2/neu was assessed using our automated quantitative analysis method. We used cytokeratin to define pixels as breast cancer (tumor mask) within the array spot, and we measured biomarker expression within the mask using Cy5 conjugated antibodies. RESULTS: High Bcl-2 expression was associated with improved survival in the entire cohort and in the node-positive subset (P = 0.008 and P = 0.002, respectively). High Bag-1 expression was associated with improved survival in the node-positive subset (P = 0.006). On multivariable analysis, neither Bcl-2 nor Bag-1 retained their independence as prognostic markers. Strong associations were found between Bag-1, Bcl-2, estrogen receptor and progesterone receptor. CONCLUSION: Bag-1 and Bcl-2 expression in breast tumors is associated with improved outcome and steroid receptor positivity. Evaluation of Bcl-2 and Bag-1 expression in breast cancer may identify a subset of patients with a favorable prognosis, who might not benefit from chemotherapy or who might benefit from Bcl 2 targeting agents in addition to antihormonal therapy. PMID- 18430252 TI - Intramuscular recurrence in a hepatocellular carcinoma patient with indolent disease course. AB - BACKGROUND: Hepatocellular carcinoma (HCC) is a common malignancy worldwide and treatment options are depended on the stage of the tumour. In general, the prognosis of HCC patients with extra-hepatic metastasis is poor. The most common sites of extra-hepatic metastasis are the lung, abdominal lymph nodes and bone. CASE PRESENTATION: Here, we reported a 54-year-old man with an indolent clinical course of HCC. He had multiple extra-hepatic recurrences after initial hepatectomy for HCC and was benefited from repeated resections with prolonged survival. Eventually, he developed intramuscular recurrence in the thigh, which was initially mistaken as deep vein thrombosis. CONCLUSION: Selected patients with indolent disease course of HCC may benefit from repeated resections of extra hepatic metastases with prolonged survival. PMID- 18430253 TI - Rebleeding rate after interventional therapy directed by capsule endoscopy in patients with obscure gastrointestinal bleeding. AB - BACKGROUND: The precise role of capsule endoscopy in the diagnostic algorithm of obscure gastrointestinal bleeding has yet to be determined. Despite the higher diagnostic yield of capsule endoscopy, the actual impact on clinical outcome remains poorly defined. The aim of this study was to evaluate the follow-up results of patients with obscure gastrointestinal bleeding to determine which management strategies after capsule endoscopy reduced rebleeding. METHODS: All patients in whom the cause of obscure gastrointestinal bleeding was investigated between May 2004 and March 2007 were studied retrospectively. We evaluated the clinical outcome of patients with obscure gastrointestinal bleeding after capsule endoscopy using the rebleeding rate as the primary outcome. RESULTS: Seventy seven patients with obscure gastrointestinal bleeding underwent capsule endoscopy. Capsule endoscopy identified clinically significant findings that were thought to be the sources of obscure gastrointestinal bleeding in 58.4% of the patients. The overall rebleeding rate was 36.4%. The rebleeding rate was significantly higher among patients with insignificant findings than among those with significant findings (p = 0.036). Among the patients in whom capsule endoscopy produced significant findings, the rebleeding rate of the patients who underwent therapeutic interventions was significantly lower than that in those who did not undergo intervention (9.5% vs 40.0%, p = 0.046). CONCLUSION: Follow up and further aggressive interventions are necessary for patients with obscure gastrointestinal bleeding and significant capsule endoscopy findings to reduce the chance of rebleeding. PMID- 18430254 TI - Identification and molecular characterisation of a fibrinogen binding protein from Streptococcus iniae. AB - BACKGROUND: Binding of serum components by surface M-related proteins, encoded by the emm genes, in streptococci constitutes a major virulence factor in this important group of organisms. The present study demonstrates fibrinogen binding by S. iniae, a Lancefield non-typeable pathogen causing devastating fish losses in the aquaculture industry and an opportunistic pathogen of humans, and identifies the proteins involved and their encoding genes. RESULTS: Fibrinogen binding by S. iniae significantly reduced respiratory burst activity of barramundi peritoneal macrophages in primary cultures compared to BSA-treated or untreated controls, indicating a potentially important role for fibrinogen binding cell-surface proteins in avoiding phagocytic attack in fish. We describe a novel emm-like gene, simA, encoding a 57 kDa fibrinogen binding M-like protein in S. iniae. These SiM proteins and their corresponding tetrameric structures from some sequevar types (approximately 230 kDa) bound fibrinogen in Western blots. simA was most closely related (32% identity) to the demA gene of S. dysgalactiae. Genome walking and sequencing determined the genetic organization of the simA region had similarities to the mgrC regulon in GCS and to S. uberis. Moreover, a putative multigene regulator, mgx was orientated in the opposite direction to the simA gene in common with S. uberis, but contrary to findings in GAS and GCS. In GAS, diversity among emm-genes and consequent diversity of their M-related proteins results in substantial antigenic variation. However, an extensive survey of S. iniae isolates from diverse geographic regions and hosts revealed only three variants of the gene, with one sequevar accounting for all but two of the 50 isolates analysed. CONCLUSION: These proteins play a role in avoiding oxidative attack by phagocytic cells during infection of fish by S. iniae, but genetic diversity amongst these key surface proteins has not yet arisen. This lack of diversity coupled with a functional role in macrophage resistance suggests that these proteins may constitute important targets for future vaccines against S. iniae in fish. PMID- 18430256 TI - Evaluation of a web-based ECG-interpretation programme for undergraduate medical students. AB - BACKGROUND: Most clinicians and teachers agree that knowledge about ECG is of importance in the medical curriculum. Students at Karolinska Institute have asked for more training in ECG-interpretation during their undergraduate studies. Clinical tutors, however, have difficulties in meeting these demands due to shortage of time. Thus, alternative ways to learn and practice ECG-interpretation are needed. Education offered via the Internet is readily available, geographically independent and flexible. Furthermore, the quality of education may increase and become more effective through a superior educational approach, improved visualization and interactivity. METHODS: A Web-based comprehensive ECG interpretation programme has been evaluated. Medical students from the sixth semester were given an optional opportunity to access the programme from the start of their course. Usage logs and an initial evaluation survey were obtained from each student. A diagnostic test was performed in order to assess the effect on skills in ECG interpretation. Students from the corresponding course, at another teaching hospital and without access to the ECG-programme but with conventional teaching of ECG served as a control group. RESULTS: 20 of the 32 students in the intervention group had tested the programme after 2 months. On a five-graded scale (1- bad to 5 - very good) they ranked the utility of a web based programme for this purpose as 4.1 and the quality of the programme software as 3.9. At the diagnostic test (maximal points 16) by the end of the 5-month course at the 6th semester the mean result for the students in the intervention group was 9.7 compared with 8.1 for the control group (p = 0.03). CONCLUSION: Students ranked the Web-based ECG-interpretation programme as a useful instrument to learn ECG. Furthermore, Internet-delivered education may be more effective than traditional teaching methods due to greater immediacy, improved visualisation and interactivity. PMID- 18430255 TI - Cell division in Escherichia coli cultures monitored at single cell resolution. AB - BACKGROUND: A fundamental characteristic of cells is the ability to divide. To date, most parameters of bacterial cultures, including cell division, have been measured as cell population averages, assuming that all bacteria divide at a uniform rate. RESULTS: We monitored the division of individual cells in Escherichia coli cultures during different growth phases. Our experiments are based on the dilution of green fluorescent protein (GFP) upon cell division, monitored by flow cytometry. The results show that the vast majority of E. coli cells in exponentially growing cultures divided uniformly. In cultures that had been in stationary phase up to four days, no cell division was observed. However, upon dilution of stationary phase culture into fresh medium, two subpopulations of cells emerged: one that started dividing and another that did not. These populations were detectable by GFP dilution and displayed different side scatter parameters in flow cytometry. Further analysis showed that bacteria in the non growing subpopulation were not dead, neither was the difference in growth capacity reducible to differences in stationary phase-specific gene expression since we observed uniform expression of several stress-related promoters. The presence of non-growing persisters, temporarily dormant bacteria that are tolerant to antibiotics, has previously been described within growing bacterial populations. Using the GFP dilution method combined with cell sorting, we showed that ampicillin lyses growing bacteria while non-growing bacteria retain viability and that some of them restart growth after the ampicillin is removed. Thus, our method enables persisters to be monitored even in liquid cultures of wild type strains in which persister formation has low frequency. CONCLUSION: In principle, the approaches developed here could be used to detect differences in cell division in response to different environmental conditions and in cultures of unicellular organisms other than E. coli. PMID- 18430257 TI - ADHD and Disruptive Behavior scores - associations with MAO-A and 5-HTT genes and with platelet MAO-B activity in adolescents. AB - BACKGROUND: Pharmacological and genetic studies suggest the importance of the dopaminergic, serotonergic, and noradrenergic systems in the pathogenesis of Attention Deficit Hyperactivity Disorder (ADHD) and Disruptive Behavior Disorder (DBD). We have, in a population-based sample, studied associations between dimensions of the ADHD/DBD phenotype and Monoamine Oxidase B (MAO-B) activity in platelets and polymorphisms in two serotonergic genes: the Monoamine Oxidase A Variable Number of Tandem Repeats (MAO-A VNTR) and the 5-Hydroxytryptamine Transporter gene-Linked Polymorphic Region (5-HTT LPR). METHODS: A population based sample of twins, with an average age of 16 years, was assessed for ADHD/DBD with a clinical interview; Kiddie Schedule for Affective Disorders and Schizophrenia for School-Age Children-Present and Lifetime Version (K-SADS-PL). Blood was drawn from 247 subjects and analyzed for platelet MAO-B activity and polymorphisms in the MAO-A and 5-HTT genes. RESULTS: We found an association in girls between low platelet MAO-B activity and symptoms of Oppositional Defiant Disorder (ODD). In girls, there was also an association between the heterozygote long/short 5-HTT LPR genotype and symptoms of conduct disorder. Furthermore the heterozygote 5-HTT LPR genotype in boys was found to be associated with symptoms of Conduct Disorder (CD). In boys, hemizygosity for the short MAO-A VNTR allele was associated with disruptive behavior. CONCLUSION: Our study suggests that the serotonin system, in addition to the dopamine system, should be further investigated when studying genetic influences on the development of Disruptive Behavior Disorders. PMID- 18430258 TI - The area-based social patterning of injuries among 10 to 19 year olds. Changes over time in the Stockholm County. AB - BACKGROUND: Area-based studies of childhood injuries strongly suggest that neighborhood socio-demographic and economic circumstances impact on various - though not all - types of injuries. The primary aim of this study was to investigate the stability over time of the association between area characteristics and childhood injuries of various causes. METHODS: Register-based and ecological, the study encompassed Stockholm County's 138 parishes, and considered two time periods (1993-95; 2003-05). Two indices were measured: economic deprivation and social fragmentation, and parishes were allocated to their respective quintile on each index. Data on both unintentional and intentional injuries for children (boys and girls) aged 10-14 and 15-19 respectively were gathered from the County Council's hospital inpatient register. For each period and index, gender, age and cause-specific comparisons were made to assess the rate ratios (with 95% confidence intervals) of being injured using parishes belonging to the best index level as a comparison group. A series of simple and partial Pearson correlations were also calculated to assess the independent contribution of each index. RESULTS: Regardless of time period, there were rather few significant rate ratios and, when they occurred, there were both under and excess risks. For instance, in each period, boys from both age groups living in parishes with the highest levels of economic deprivation had lower rate of injury as a motor vehicle rider. Most strikingly, intentional injuries were more frequent during the second time period and in considerable excess among girls aged 15-19 from more economically deprived areas. Also, during that last period, none of the injury causes correlated significantly with the index of social fragmentation after adjustment for economic deprivation (partial correlation). CONCLUSION: Over a ten-year period, differential economic deprivation among parishes has widened more than social fragmentation in Stockholm County. The correlation between those indices is high in both periods of time whilst the association between the levels of each index and injury rates varies depending on group of injuries or time period considered. It is of concern that intentional injuries have increased numerically and are significantly and positively correlated with economic deprivation (net of social fragmentation), in particular among girls. PMID- 18430260 TI - NHERF1: molecular brake on the PI3K pathway in breast cancer. AB - The adaptor protein NHERF1/EBP50 (Na/H exchanger regulatory factor 1/ezrin radixin-moesin-binding phosphoprotein 50) emerged recently as an important player in breast cancer progression. Consisting of two tandem PDZ domains linked to a carboxyl-terminal ezrin-binding region, NHERF1 assembles macromolecular complexes at the apical membrane of epithelial cells in many epithelial tissues, including the mammary gland. Involved initially in trafficking and regulation of transmembrane ion transporters and G protein-coupled receptors, NHERF1 also couples molecules involved in cell growth, such as the platelet-derived growth factor receptor (PDGFR) and PTEN (phosphatase and tensin homolog deleted on chromosome 10). In the previous issue of Breast Cancer Research, Pan and colleagues show an inhibitory action of NHERF1 on the phosphoinositide-3 kinase (PI3K)/Akt pathway in breast cancer cells via interaction of NHERF1 with PTEN, the physiological antagonist of the PI3K. Additionally, they show that NHERF1 expression confers susceptibility to PDGFR pharmacological inhibition depending on the presence of PTEN tumor suppressor. PMID- 18430261 TI - Adult cystic hygroma: successful use of OK-432 (Picibanil). AB - OBJECTIVE: We report an adult case of cystic lymphangioma treated with OK-432 (Picibanil). METHOD: A case report and review of the literature concerning the use of OK-432 to treat cystic lymphangioma is presented. RESULTS: A 31-year-old woman developed a cystic lymphangioma four weeks post-partum. This was treated initially by aspiration, for diagnostic purposes. Investigation suggested that surgery would be challenging. A review of the literature demonstrated success with OK-432 in the treatment of this condition, although primarily in the paediatric population. This patient was successfully treated thus, and at the time of writing remained symptom free. A suggested management plan is outlined. CONCLUSION: Treatment with OK-432 is useful in the management of cystic lymphangiomas in adults and should be considered as first line treatment. PMID- 18430262 TI - Effect of the form of the sesame-based diet on the absorption of lignans. AB - The effect of different forms of sesame-based diets on the concentration of plasma lignans was assayed by estimating the levels of certain lignans (sesame lignans and enterolignans) in the plasma of experimental animals. In a series of experiments, male Wistar rats were fed either a raw sesame-enriched diet or a tahini-enriched diet. The plasma concentration of the lignans (sesame lignans and enterolignans) was determined at various time intervals over a 24 h period after a single administration. Enterodiol and enterolactone concentration in the tahini treated group was significantly higher than in the raw sesame-treated group. In another series of experiments, male Wistar rats were fed, for 15 d, diets enriched in raw dehulled sesame, sesame perisperm, sesame oil, tahini and a polyphenolic extract derived from the seed perisperm. Enterodiol and enterolactone plasma concentration was high in the case of the sesame perisperm in spite of its low concentration in the assessed sesame lignans. Overall, the levels of the sesame lignans and enterolignans present in plasma seem to be influenced not only by the amount of lignan intake but also by other factors such as the form of the sesame-based diet. PMID- 18430259 TI - Systemic 5-fluorouracil treatment causes a syndrome of delayed myelin destruction in the central nervous system. AB - BACKGROUND: Cancer treatment with a variety of chemotherapeutic agents often is associated with delayed adverse neurological consequences. Despite their clinical importance, almost nothing is known about the basis for such effects. It is not even known whether the occurrence of delayed adverse effects requires exposure to multiple chemotherapeutic agents, the presence of both chemotherapeutic agents and the body's own response to cancer, prolonged damage to the blood-brain barrier, inflammation or other such changes. Nor are there any animal models that could enable the study of this important problem. RESULTS: We found that clinically relevant concentrations of 5-fluorouracil (5-FU; a widely used chemotherapeutic agent) were toxic for both central nervous system (CNS) progenitor cells and non-dividing oligodendrocytes in vitro and in vivo. Short term systemic administration of 5-FU caused both acute CNS damage and a syndrome of progressively worsening delayed damage to myelinated tracts of the CNS associated with altered transcriptional regulation in oligodendrocytes and extensive myelin pathology. Functional analysis also provided the first demonstration of delayed effects of chemotherapy on the latency of impulse conduction in the auditory system, offering the possibility of non-invasive analysis of myelin damage associated with cancer treatment. CONCLUSIONS: Our studies demonstrate that systemic treatment with a single chemotherapeutic agent, 5-FU, is sufficient to cause a syndrome of delayed CNS damage and provide the first animal model of delayed damage to white-matter tracts of individuals treated with systemic chemotherapy. Unlike that caused by local irradiation, the degeneration caused by 5-FU treatment did not correlate with either chronic inflammation or extensive vascular damage and appears to represent a new class of delayed degenerative damage in the CNS. PMID- 18430264 TI - Outbreaks of noroviral gastroenteritis in Florida, 2006-2007. AB - Noroviruses are an important cause of sporadic cases and outbreaks of acute gastroenteritis. During 2006-2007, widespread increases in acute gastroenteritis outbreaks consistent with norovirus were observed in the United States. We conducted a statewide survey to characterize norovirus outbreak activity in Florida during a 1-year period. From July 2006 to June 2007, 257 outbreaks of norovirus gastroenteritis were identified in 39 of Florida's 67 counties. About 44% of outbreaks were laboratory confirmed as norovirus and 93% of these were due to genogroup GII. About 63% of outbreaks occurred in long-term care facilities and 10% of outbreaks were classified as foodborne. The median number of ill persons per outbreak was 24, with an estimated total of 7880 ill persons. During the study period, norovirus outbreak activity in Florida was widespread, persistent, and consistent with increased activity observed in other parts of the country. PMID- 18430263 TI - Racial and seasonal differences in 25-hydroxyvitamin D detected in maternal sera frozen for over 40 years. AB - Serum banks from large, decades-old epidemiological studies provide a valuable opportunity to explore the contributions of in utero vitamin D exposure to fetal origins of adult diseases. We compared 25-hydroxyvitamin D (25(OH)D) by race and season (two powerful predictors of vitamin D status) in sera frozen for >or= 40 years with sera frozen for 150 ng/mL. Three specimens that screened negative, but produced absorbance values that were intermediate between negative and positive controls, were submitted for GC-MS analysis; BE was detected in all three specimens at concentrations of 54, 94, and 119 ng/mL. Twenty-eight specimens produced screening results indistinguishable from negative controls. Within the limitations of the study design, we conclude that amoxicillin is unlikely to produce false-positive urine screens for cocaine metabolites. PMID- 18430301 TI - Evidence that morphine is metabolized to hydromorphone but not to oxymorphone. AB - A minor pathway for the biotransformation of morphine to hydromorphone has been identified in humans. Recently, an unsubstantiated claim that morphine is metabolized to hydromorphone and then to oxymorphone was published. The goal of this study was to determine if credible evidence that oxymorphone is a metabolite of either morphine or hydromorphone exists. Urine specimens from pain patients who were treated exclusively with high daily doses of morphine (N = 34) or hydromorphone (N = 26) were analyzed by liquid chromatography-tandem mass spectrometry for oxymorphone, hydromorphone, and morphine (LOD = 25 ng/mL). Specimens were also tested for a variety of other medications. Criteria for inclusion of patients' specimens were as follows: 1. patients were undergoing exclusive dosing with either morphine or hydromorphone; 2. non-prescribed medications were not detected; and 3. urine concentrations of morphine were > 100,000 ng/mL for the high-dose morphine group and > 1,000 ng/mL of hydromorphone for the high-dose hydromorphone group. Consistent with earlier reports, hydromorphone was detected in patients treated with high-dose morphine. The ratio of hydromorphone to morphine ranged from 0.2 to 2.2%. Oxymorphone was not detected in any specimen from high-dose morphine or high-dose hydromorphone patients. The authors conclude, based on these data, that oxymorphone is not a metabolite of morphine or hydromorphone. PMID- 18430302 TI - A fatality involving 1,3-propanediol and its implications in measurement of other glycols. AB - The decomposed body of a 45-year-old female was found, face down, in a mobile home, along with a suicide note and two antifreeze containers. Analysis of the body fluid collected from the decedent showed the presence of 58 mg/dL ethanol, but suspected ethylene glycol was not found in the sample. However, an unusually large peak of internal standard, 1,3-propanediol, was found in the sample. Gas chromatography-mass spectrometry analysis confirmed the presence of 1,3 propanediol in the sample. Using gas chromatography-flame-ionization detection, the concentration of 1,3-propanediol was determined to be 445 mg/dL. To our knowledge, this is the first report involving 1,3-propanediol as the cause of death. The study also highlights the importance for the close scrutiny of data, as 1,3-propanediol is a frequently used internal standard for the assay of glycols. PMID- 18430303 TI - Hirsch-index for winners of TIAFT's mid-career achievement award. PMID- 18430304 TI - In Memoriam: Donald C. Fournier, M.D. (1949 - 2008). PMID- 18430305 TI - Is asthma in childhood different from asthma in adults? Why do we need special approaches to asthma in children? AB - Childhood asthma poses a number of challenges. In many asthmatic patients, the disease begins in infancy or early childhood and aeroallergen sensitization in the first 3 years of life appears to dictate later outcomes. The airway manifestations of asthma in childhood differ from those in adults and require different therapeutic approaches. Importantly, corticosteroid treatment of childhood asthma appears to have little impact on the progressive loss of lung function in a subset of patients. PMID- 18430306 TI - The future of the long-acting beta-adrenergic bronchodilators in the treatment of asthma. AB - The combination of long-acting beta-agonist (LABA) bronchodilators with inhaled corticosteroids (ICSs) has been shown to be an extremely effective treatment for asthma. Use of LABA as monotherapy for asthma is associated with increased adverse events including exacerbations and asthma deaths. However, intensive evaluation of the combined LABA-ICS therapy provided no signals of increased risk. LABA appears to potentiate the effects of ICS. This provides the opportunity for use a of lower ICS dose for asthma control with less risk of steroid side effects. The combination of formoterol and budesonide used as both maintenance and relief medications may offer superior asthma control with less medication use. The recent introduction of 24-hour LABA, which are in clinical trials, makes possible the concept of very effective once-daily combinations of LABA and ICS, which would be expected to increase patient adherence and improve asthma outcomes. The 24-hour LABA will likely be combined with a 24-hour anticholinergic to treat chronic obstructive pulmonary disease. Whether this dual combination with ICS will enhance our treatment of more severe asthma remains an exciting hypothesis to be tested. PMID- 18430307 TI - Pharmacoeconomic review of medical management of persistent asthma. AB - Asthma affects 20 million Americans and causes a substantial loss of productivity. Medications help to increase symptom-free days and improve quality of life. Examining the cost-effectiveness of different treatments, in addition to their clinical efficacy, allows us to choose the optimal strategy in managing patients. This study reviews published pharmacoeconomic analyses of different medications used for asthma management, with a focus on medications available in the United States. English language, peer-reviewed articles, or abstracts were identified from MEDLINE and Current Contents databases (both 1966 to March 1, 2006) using the search terms asthma, pharmacoeconomics, cost-effectiveness, steroids, beta(2)-agonists, cromolyn, methylxanthines, leukotriene receptor antagonists, and omalizumab. Citations from available articles were reviewed also for additional references. Pharmacoeconomic analysis from a payer's perspective has shown that salmeterol/fluticasone is a cost-effective treatment option for moderate persistent asthma management, when compared with fluticasone with or without the addition of leukotriene modifiers. Leukotriene modifiers are less cost-effective than inhaled corticosteroids or combined inhaled steroids and long acting beta(2)-agonists for mild or moderate persistent asthma. Anti-IgE antibody has been shown inconsistently, to be cost-effective in patients with moderate to severe allergic asthma. Although the acquisition cost of levalbuterol is higher, one study showed that it may be more cost-effective than albuterol after taking into account reduction in hospitalizations. Cost-effectiveness analyses and clinical efficacy of medications, together with other patient-specific factors, are important information to be considered when selecting treatment regimens for asthma. Future economic analysis should focus on finding better ways to evaluate productivity lost due to asthma, in addition to hospitalization. PMID- 18430308 TI - Harmonia axyridis ladybug invasion and allergy. AB - Beginning in 1916 Harmonia axyridis, an orange/red lady beetle with variable black spotting, was imported into the United States from Asia. This agricultural pest-control predator established independent feral populations in North America by 1988. Subsequently, Harmonia axyridis has become a pest to homeowners and various horticultural enterprises. Seeking winter hibernation sites, ladybug swarms invade human homes/habitats primarily in the fall. With increased Harmonia axyridis exposures, human ladybug allergy was first reported in 1998. Ladybug specific IgE hypersensitivity has been reported in all ages (1-78 years old) and both sexes. Clinical ladybug allergy manifests variously as rhinoconjunctivitis, asthma, urticaria, and angioedema. A majority, but not all, allergic individuals are primarily exposed at home. Large fall swarms and smaller spring dispersions produce corresponding peaks in ladybug allergy. Ladybug hemolymph is a primary source of allergen. Har a 1 and Har a 2 major ladybug allergens have been characterized. Ladybug allergy prevalence in one endemic area was reported as 10%. Self-report of ladybug pests at home did not predict ladybug allergy, suggesting other exposures are important also. Some individuals have no history of atopy before manifestation of ladybug allergy. Ladybug, cat, cockroach, and house-dust mites are the most likely allergens to present as isolated single positive skin tests in an allergist's office. Ladybug should be a standard skin test allergen for all allergy patients tested in endemic areas. Avoidance of ladybug exposure is paramount to treatment. PMID- 18430309 TI - Sublingual immunotherapy: dosing in relation to clinical and immunological efficacy. AB - An increasing number of clinical immunotherapy trials have been published in the United States and Europe. In the allergy community on both sides of the Atlantic, there is a growing interest in obtaining a better understanding of the allergen doses used in these studies. The question of effective allergen dose is particularly important in sublingual immunotherapy, but the effective dose range still is to be determined. PMID- 18430310 TI - The role of montelukast on perennial allergic rhinitis and associated sleep disturbances and daytime somnolence. AB - Perennial allergic rhinitis (PAR) often causes sleep disturbances and associated daytime somnolence, thus resulting in a poor quality of life. Various clinical interventions in patients suffering from the disorder seek to improve symptoms and quality of life. Additional studies are needed to establish whether the alleviation of PAR symptoms, particularly the reduction of congestion, will improve sleep quality and reduce daytime somnolence. This study seeks to determine whether treatment with montelukast is more effective than placebo in reducing nasal congestion and sleep disturbances, resulting in reduced daytime somnolence and fatigue in patients with PAR. Thirty-one subjects were enrolled in a double-blinded, placebo-controlled study using Balaam's design. Patients were treated with montelukast or placebo. Collected subjective data included a daily diary recording nasal symptoms, sleep issues, and daytime fatigue, the Functional Outcomes of Sleep Questionnaire, the Epworth Sleepiness Scale, Juniper's Rhinoconjunctivitis Quality of Life Questionnaire, the Rhinitis Severity Scale, the Calgary Sleep Apnea Quality of Life Index, and Trail Making tests. Subjects treated with montelukast, compared with placebo, showed a statistically significant improvement in daytime somnolence (p = 0.0089) and daytime fatigue (p = 0.0087), with both factors improving with montelukast and worsening with placebo. In a small cohort of subjects, montelukast, when compared with placebo, improved the symptoms of PAR and reduced the fatigue and daytime somnolence associated with the disorder. PMID- 18430311 TI - Mediator measurements after daily instillation of allergen: Increased IL-5 and decreased IFN-gamma. AB - This study was designed to measure symptoms, IL-4, IL-5, IFN-gamma, and eosinophilic cationic protein (ECP) in nasal secretions from subjects experiencing an artificial allergy season and to look for evidence of priming. Clinically relevant allergen was administered intranasally out of season to 12 asymptomatic individuals with seasonal allergic rhinitis. These individuals were then randomized to receive allergen or saline daily for the next 7 days. Nasal secretions and scrapings of nasal epithelium were obtained at baseline (day 1), 24 hours after the initial allergen administration (day 2), and 24 hours after the last instillation of allergen or saline (day 9). Nasal symptom scores (p < 0.0002), IL-5 mRNA (p = 0.03), and ECP (p < 0.02) increased after receiving the first challenge (day 2 compared with day 1). In the six subjects randomized to receive seven sequential daily challenges with allergen, symptom scores remained elevated (p < 0.02), IL-5 protein increased (p = 0.02), and IFN-gamma (p = 0.02) levels decreased (day 9 compared with day 1). In the six subjects randomized to receive seven sequential daily challenges with placebo, symptom scores, IL-5, and IFN-gamma levels were not significantly different (day 9 compared with day 1). Compared with the findings at day 2 (n = 12), the treated subjects (n = 6) had no further increase in symptoms but did show a further increase in IL-5 (p = 0.01) and a decrease in IFN-gamma (p = 0.02) at day 9. Daily instillation of moderate doses of allergen intranasally is characterized by persistent symptoms, elevation of IL-5, and reduced levels of IFN-gamma. PMID- 18430312 TI - Correlation between specific IgA and eosinophil numbers in the lavage fluid of patients with perennial allergic rhinitis. AB - The mechanisms of eosinophil activation in perennial allergic rhinitis (AR) are not fully understood but may be mediated by a series of inflammatory mediators, including locally produced specific antibodies. The aim of this study was to evaluate the role of locally produced allergen-specific antibodies in the nasal lavage fluid during the recruitment of inflammatory cells in AR. Thirteen patients with AR and a positive nasal provocation test (NPT) with Dermatophagoides pteronyssinus and 10 asymptomatic subjects, sensitized to D. pteronyssinus but who had a negative NPT were enrolled. Nasal lavage fluids were collected before and at 10, 30, and 60 minutes and 3, 6, and 24 hours after the NPT. Inflammatory cells were counted, and eosinophil cationic protein (ECP) and specific immunoglobulin E (IgE), IgG, and IgA antibodies were measured by enzyme linked immunosorbent assay. After NPT, six subjects showed early responses and seven subjects showed dual responses. The eosinophil count and ECP level in lavage fluid were significantly increased after NPT in patients with positive NPTs, and these values were significantly higher than in the negative group. The specific IgA level in lavage fluid was increased after NPT in the positive group, and significant differences were noted for both the early and the late positive responses. Significant correlations were found between specific IgA and ECP levels in lavage fluid during early and late responses. Eosinophil recruitment occurred during both early and late responses after allergen exposure in perennial AR. Locally produced allergen-specific IgA (but not IgE or IgG) may be involved in eosinophil activation in both early and late responses. PMID- 18430313 TI - Risk factors associated with transient wheezing in young children. AB - Transient wheezing in young children has been reported to be independent of atopy. Although persistence of early wheezing has been associated with factors related to allergy in multiple studies, transient wheezing has not been similarly studied. The Childhood Allergy Study birth cohort was the source of these data. Transient wheezing was defined as history of wheezing in the past 12 months at ages 1, 2, and/or 4 years, but not at 6 years, and evaluated in relationship to aeroallergen-specific circulating IgE and positive skin testing as markers of an atopic profile. Testing for IgE and skin-prick testing to dust mites, dogs, cats, ragweed, and timothy were performed at the age of 6 years. Other variables in logistic regression analyses were sex; breast-feeding; birth order; parental allergy and smoking history; and household pets, daycare, fever, and antibiotic use in the 1st year of life. Of 372 children, 128 (34.4%) experienced transient wheezing and 175 (47.0%) never wheezed. Atopy was not associated with transient wheezing (adjusted odds ratio for a positive allergen-specific IgE test = 1.2, p = 0.66; skin-prick test = 0.8, p = 0.47). Boys were more likely to be transient wheezers (adjusted relative risk [RR] = 1.7; 95% confidence interval [CI], 1.1 2.8; p = 0.018). Transient wheeze was associated with antibiotic treatment in the first 6 months of life (adjusted RR = 1.6; 95% CI, 1.0-2.6; p = 0.048). We confirm previous observations that transient wheezing in young children is not associated with an atopic predisposition. PMID- 18430314 TI - Prevalence of atopy in children with type 1 diabetes mellitus, hepatitis B virus carriers, and healthy children: role of T helper 1 (Th1)-type immune response. AB - The prevalence of allergic diseases such as asthma, hay fever, and atopic dermatitis has increased over the past few decades, especially in developed countries. They are characterized by a chronic inflammatory reaction mediated by T helper 2 (Th2) cells. Two common chronic diseases of childhood-an autoimmune disease, type 1 diabetes mellitus (DM), and a chronic viral infection, hepatitis B virus (HBV) carriers-are associated with a Th1-dominant and Th1-insufficient cytokine profile, respectively. The purpose of this study was to analyze the frequency of allergic disease in patients with type 1 DM and, in HBV carriers, to evaluate the role of Th1-type immune response in atopy and allergic disease. The study included patients with type 1 DM (group I, n = 52), HBV carriers (group III, n = 47), and a healthy control group (group III, n = 209). Participants were screened for allergic disease and atopic sensitization. Symptoms of asthma, eczema, and atopy were found more commonly in HBV carrier children compared with those with DM and healthy controls. This study supports the Th1/Th2 model. The prevalence of allergic disease and atopy is decreased in Th1-mediated autoimmune disease, type 1 DM, and, conversely, is increased in insufficient Th1 response, chronic HBV carriers. Additional studies are needed to evaluate the effect of atopy and allergic diseases in glycemic control and long-term complications in patients with type 1 DM and the effect of atopy on progression of chronic HBV infection. PMID- 18430315 TI - Exhaled nitric oxide levels during treatment in patients hospitalized with asthma. AB - The time required for fractional excretion of nitric oxide (FE(NO)) measurements to acutely change after systemic corticosteroids is unknown, limiting the usefulness of this biomarker in hospital treatment and discharge decisions. The purpose of this study was to follow FE(NO) measurements of hospitalized adult patients with asthma receiving therapy and to correlate FE(NO) with forced expiratory volume in 1 second percent predicted (FEV(1)%). Ten acute asthmatic patients who required hospitalization were recruited and treated with standard therapy. FE(NO) measurements were performed at presentation to the emergency department (baseline), as well as 1, 4, 6, 8, 12, and 24 hours after the initiation of therapy. FEV(1)% was measured at baseline, 1, 6, 12, and 24 hours. Subjects also were called 3 days after discharge to assess if symptoms had improved. The baseline FE(NO) was 57.5 parts per billion (ppb). There was no significant change over the first 8 hours. At 12 hours, there was an increase to 96.5 ppb (p = 0.01). Compared with baseline, all 10 subjects showed an increase at the 12-hour time point, with an average increase of 52%. The correlation between change in FE(NO) and change in FEV(1)% approached significance (p = 0.089). Subjects who improved after discharge had a greater percent increase in FE(NO) than those who did not (p = 0.043). FE(NO) measurements increase in hospitalized asthmatic patients receiving therapy. This augmentation appears to be associated with improvements in FEV(1). Asthmatic patients who show a greater increase in FE(NO) may have better outcomes after discharge. PMID- 18430316 TI - Health care utilization and costs of adult asthma in Taiwan. AB - The burden of asthma is sufficient to warrant its recognition as a high-priority disorder in governmental health strategies in many countries. However, the components of the total health care costs for patients with asthma have not been well studied, and an overall understanding of health care utilization patterns in this population is lacking in Taiwan. We evaluated 95,110 patients aged 18-55 years who were enrolled in the National Health Insurance Research Database from January 1 to December 31, 2002. Health care utilization and costs, including those related to office, outpatient hospital, emergency department, and inpatient hospital visits were compared between patients with and patients without asthma. In 2002, the period prevalence of treated asthma was 1.8%. Patients with asthma used substantially more services than did those without asthma in all categories. The mean costs of hospitalizations for patients with asthma were 2.7-fold higher than those of patients without asthma. Asthma care represented 26% of all health care services that patients with asthma received, while the remaining 74% were for nonasthma care. Almost one-half of all asthma-related costs were attributable to hospitalizations. These findings may serve as baseline data for future evaluations of changes in health care utilization and expenditures among asthmatic patients. PMID- 18430317 TI - The effect of enantiomers of beta-agonists on myofibroblast-derived vascular endothelial growth factor and other matrix components in the presence of dust mite extract. AB - Beta(2)-adrenergic receptor agonists have been shown to modulate airway epithelial cell and smooth muscle release of cytokines and growth factors transforming growth factor (TGF) beta, associated with remodeling, is known to up regulate the synthesis of vascular endothelial growth factor (VEGF) and stimulate differentiation of fibroblasts to the myofibroblast phenotype. VEGF and fibronectin can promote angiogenesis and (S)-albuterol can induce VEGF secretion from normal human lung fibroblasts (NHLF). We hypothesize that (S)-albuterol could stimulate myofibroblast secretion and expression of VEGF and fibronectin in the presence of Dermatophagoides pteronyssinus extract. Cultured NHLFs were stimulated with IL-1beta, TGF-beta, D. pteronyssinus, and treated with (R)- and (S)-enantiomers of albuterol. VEGF and fibronectin and basic fibroblast growth factor (bFGF) were measured by ELISA and mRNA. VEGF secretion by fibroblasts was twofold higher with 10(-7) M of (R) relative to (S) (p < 0.05). Myofibroblast secretion of VEGF was increased twofold over fibroblasts, but there was no difference between enantiomers. (S)-albuterol at 10(-8)-10(-4) M caused an increase in VEGF mRNA that paralleled VEGF secretion relative to 10(-8)-10(-4) M. Fibronection secretion by myofibroblasts but not fibroblasts was increased by 10( 5) M of (S) relative to (R) in the presence of recombinant interleukin 1 (rhIL 1)beta and D. pteronyssinus (S)-albuterol at 10(-6) M increased bFGF. The 10(-6) M of (S)-albuterol, but not (R)-albuterol, may promote angiogenesis. Increased fibronectin or bFGF by (S)-albuterol could enhance matrix deposition and remodeling in a subset of asthmatic patients. PMID- 18430318 TI - Role of regular treatment with inhaled corticosteroid or leukotriene receptor antagonist in mild intermittent asthma. AB - Current guidelines for asthma treatment do not recommend daily maintenance therapy in patients with mild intermittent (step 1) asthma. However, because there is increasing evidence that airway inflammation is present even in this patient group, maintenance anti-inflammatory therapy may be considered. We investigated the clinical impact of regular treatment with the inhaled corticosteroid beclomethasone dipropionate and the leukotriene receptor antagonist pranlukast in the patients concerned. The study was a randomized, controlled, parallel-group, multicenter trial. Eighty-five symptomatic patients with newly diagnosed mild intermittent asthma having normal pulmonary function were assigned beclomethasone or pranlukast for 8 weeks. Then, these medications were stopped for the next 16 weeks. Main outcome measures were asthma symptoms, pulmonary function, and airway inflammation. Treatment with beclomethasone and pranlukast significantly increased forced expiratory volume in 1 second and peak expiratory flow from baseline and decreased asthma symptom scores and sputum eosinophil counts and eosinophil cationic protein contents. After discontinuation of the treatment, symptom scores remained unchanged, but pulmonary function and airway inflammation were aggravated and then returned to the baseline levels. Therefore, maintenance therapy with inhaled corticosteroid or leukotriene receptor antagonist can provide further improvements in asthma symptoms, pulmonary function, and airway inflammation, and discontinuation of the therapy causes worsening of asthma, indicating that stopping or interrupting anti inflammatory therapy may not be advisable in patients with symptomatic mild intermittent asthma. PMID- 18430319 TI - Respiratory syncytial virus infection in a Sicilian pediatric population: risk factors, epidemiology, and severity. AB - Respiratory syncytial virus (RSV) is the leading cause of hospitalization for lower respiratory tract infections (LRTIs) in young children worldwide. This study evaluated the epidemiological and clinical patterns of RSV infection in infants hospitalized for LRTIs in Sicily. Over a 7-month period (October 1, 2005 to April 30, 2006), all children <2 years of age hospitalized for LRTIs were evaluated and tested for respiratory viruses. Logistic regression was used to identify the risk factors associated with RSV infection and with more severe disease. One hundred sixty-four children were enrolled and 40.9% were found to be RSV(+). The epidemic peak of RSV occurred in April, and no cases were observed in October, November, and December. RSV Infections had the highest incidence in children <3 months of age (54.7%). The likelihood to be RSV(+) rather than RSV(-) was lower for female gender and children >6 months old, with a gestational age (GA) of >36 weeks, with a birth weight of >2.50 g, with previous hospitalizations due to LRTI, with smokers in the household, and with a history of breast-feeding (p < 0.05 for each). RSV infection was associated with a higher likelihood to be admitted to neonatal intensive care units and to longer hospitalizations (p = 0.061). The collected data show that, in Sicily, RSV is an important cause of LRTIs in infants and a variety of factors, such as gender, chronological age at hospitalization, GA, birth weight, and exposure to tobacco smoke and breast feeding may affect the prevalence of RSV-related lower respiratory tract disease and, possibly, the risk of developing asthma-like symptoms during the school years. PMID- 18430320 TI - Epinephrine auto-injector prescriptions as a reflection of the pattern of anaphylaxis in an Asian population. AB - This study surveyed the prescription patterns of adrenaline auto-injectors (AAs) in Singapore to examine the frequency, triggers, and demographic pattern of anaphylaxis requiring such prescriptions. A 6-year retrospective review of 417 consecutive patients prescribed AAs in Singapore from January 1999 to December 2004, as identified from hospital pharmacy records. There were 417 patients identified, consisting of 295 (70.7%) Singaporeans with the remaining being non Singaporean residents. Based on population census, the frequency of AA prescriptions was estimated at 1 per 10,000 Singaporeans. Demographic factors associated with AA prescriptions were male gender (OR = 1.361; p = 0.002); minority ethnic groups, which included Eurasians, Caucasians, Koreans, and Japanese (OR = 15.873; p < 0.001); and children <15 years of age (OR = 2.593; p < 0.001). The most common food allergens resulting in AA prescriptions were peanut (41.9%) and shellfish allergy (28.5%). Multiple logistic regression analysis showed that peanut allergy was independently associated with Eurasian ethnicity (OR = 5.045; p = 0.021); and shellfish allergy with Indian ethnicity (OR = 2.757; p = 0.034). The estimated frequency of AA prescriptions in Singapore is relatively low at 0.01%. The incidence of peanut and shellfish allergy in the Asian population appears to differ from that seen in Western populations. PMID- 18430321 TI - A boy with fever, lymphadenopathy, hepatosplenomegaly, and lymphocytosis. AB - Proliferation of the lymphoid system should arouse suspicion of a potentially serious illness. We present a 4.5-year-old boy who developed fever, vomiting, diarrhea, lymphadenopathy, hepatosplenomegaly, lymphocytosis, anemia, thrombocytopenia, and increased liver enzymes. Lymph node and bone marrow biopsies showed lymphoproliferation, Epstein-Barr virus (EBV) infection, and hemophagocytosis leading to the diagnosis of hemophagocytic lymphohistiocytosis (HLH). Chemotherapy was initiated for HLH with dexamethasone, etoposide, and cyclosporine. Because of a high level of EBV viremia, rituximab was added a few days later and resulted in a remarkable drop in the EBV in the circulation but not in the cerebrospinal fluid. However, the patient succumbed to encephalitis, pneumonia, and cardiopulmonary failure. Autopsy revealed the presence of EBV in the brain, indicating the ineffectiveness of rituximab therapy in treating central nervous system infection with EBV. PMID- 18430322 TI - Abstracts. ART Paving the Way for New Frontiers Celebrating 3 Decades of ART, 11 13 April 2008. 2nd Congress of the Asia Pacific Initiative on Reproduction and 6th Biennial Meeting of the Pacific Rim Society for Fertility & Sterility. PMID- 18430325 TI - Late presentation of a type III axis fracture with spondyloptosis. AB - A 58-year-old man presented with an undiagnosed Effendi type III classification fracture and spondyloptosis of the axis with remarkably normal neurology. We discuss his surgery 4 years since the initial injury, and the presentation, features and management of fractures of the axis. PMID- 18430326 TI - Spontaneous bilateral olecranon fractures in a rheumatoid patient. AB - A case is described of a patient sustaining bilateral spontaneous olecranon fractures while undergoing rehabilitation following surgery for a hip fracture. The patient's underlying rheumatoid arthritis disease process most likely caused erosions and subchondral cysts formation in the mid-trochlear notch region. This area, in turn, acts as a pivot point in elbow extension/flexion movements. During rehabilitation, the patient became dependent on her upper limbs for mobilisation. This led to significant forces being put through this already weakened region, causing spontaneous bilateral olecranon fractures. Rheumatoid patients after lower limb surgery should be careful during rehabilitation, and avoid undue strains through their upper limbs. PMID- 18430327 TI - Isolated calcaneal metastasis from prostate cancer. AB - We report the first case of an isolated calcaneal metastasis from prostate cancer. This case highlights the importance of considering metastatic disease in patients with persistent bone pain in the presence of known cancer, however unusual the site. PMID- 18430328 TI - A hard object in the right iliac fossa. AB - An unusual case of a Harrington rod migrating out of the abdominal cavity. PMID- 18430329 TI - Choriocarcinoma of the spleen--a rare cause of atraumatic rupture. AB - This case report identifies a very rare disease, choriocarcinoma in a male patient, causing another rare entity, an atraumatic rupture of the spleen. PMID- 18430330 TI - Emergency management of malignant acute left-sided colonic obstruction. AB - INTRODUCTION: The management of acute left-sided colonic obstruction still remains a challenging problem despite significant progress. METHODS: A literature search was undertaken using PubMed and the Cochrane Library regarding the options in emergency management of left-sided colonic obstruction focusing on outcomes such as mortality, morbidity, long-term prognosis and cost effectiveness. DISCUSSION: Colonic stenting is the best option either for palliation or as a bridge to surgery. It reduces morbidity and mortality rate and the need for colostomy formation. Stenting is likely to be cost effective, but data are variable depending on the individual healthcare system. Nevertheless, surgical management remains relevant as colonic stenting has a small rate of failure, and it is not always available. There are various surgical options. One-stage primary resection and anastomosis is the preferred choice for low-risk patients. Intra operative colonic irrigation has no proven benefit. Subtotal colectomy is useful in cases of proximal bowel damage or synchronous tumours. Hartmann's procedure should be reserved for high-risk patients. Simple colostomy has no role other than for use in very ill patients who are not fit for any other procedure. PMID- 18430331 TI - Who should run breast clinics, surgeons or radiologists? PMID- 18430332 TI - Survey of general paediatric surgery provision in England, Wales and Northern Ireland. AB - INTRODUCTION: A survey was carried out to ascertain the current provision of general paediatric surgery (GPS) in all hospitals in England, Wales and Northern Ireland with 100% return rate. The provision of GPS is at a crossroads with a drift of these cases to the overstretched, tertiary referral hospitals. METHODS: The regional representatives on the council of the Association of Surgeons of Great Britain and Ireland (ASGBI) obtained data from their regions. Any gaps in the data were completed by the author telephoning the remaining hospitals to ascertain their current provision. RESULTS: A total of 325 acute hospitals are potentially available to admit elective and/or emergency paediatric patients, of which 25 hospitals provide a tertiary paediatric surgical service. Of the remaining 'non-tertiary' hospitals, 138 provide elective GPS and 147 provide emergency GPS. The ages at which GPS is carried out varies considerably, but 76% of non-tertiary hospitals provide elective GPS to those over the age of 2 years. The ages of emergency cases are 24% over the age of 2 years and 51.5% over the age of 5 years. The age at which surgery is carried out is dependent on the anaesthetic provision. Subspecialisation within each hospital has taken place with a limited number of surgeons providing the elective surgery. 'Hub-and-spoke' provision of GPS to a district general hospital (DGH) from a tertiary centre is embryonic with only 11 surgeons currently in post. An estimate of the annual elective case load of GPS based on the average number of cases done on an operation list works out at 23,000 cases done out with the tertiary centres. DISCUSSION: Almost 10 years ago, a change in the training of young surgeons took place. An increase in training posts in Tertiary centres was made available following advice from the British Association of Paediatric Surgeons (BAPS) but these posts were often not taken up. Many DGH surgeons became uncertain whether they should continue GPS training. A subtle change in the wording of the general guidance by the Royal College of Anaesthetists altered the emphasis on the age at which it was appropriate to anaesthetise children. Change in clinical practice, reducing need, and a drift towards tertiary centres has reduced DGH operations by 30% over a decade. Young surgeons are now seldom exposed to this surgery, and are not being trained in it. The large volume of these low-risk operations in well children cannot be absorbed into the current tertiary centres due to pressure on beds. The future provision of this surgery is at risk unless action is taken now. This survey was carried out to inform the debate, and to make recommendations for the future. The principal recommendations are that: (i) GPS should continue to be provided as at present in those DGHs equipped to do so; (ii) GPS training should be carried out in the DGHs where a high volume of cases is carried out; (iii) management of these cases should use a network approach in each region; (iv) hospital trusts should actively advertise for an interest in GPS as a second subspecialty; and (v) the SAC in general surgery develop a strategy to make GPS relevant to trainee surgeons. PMID- 18430333 TI - Patient decision aids in joint replacement surgery: a literature review and an opinion survey of consultant orthopaedic surgeons. AB - INTRODUCTION: Patient decision aids could facilitate shared decision-making in joint replacement surgery. However, patient decision aids are not routinely used in this setting. METHODS: With a view to developing a patient decision aid for UK hip/knee joint replacement practice, we undertook a systematic search of the literature for evidence on the use of shared decision-making and patient decision aids in orthopaedics, and a national survey of consultant orthopaedic surgeons on the potential acceptability and feasibility of patient decision aids. RESULTS: We found little published evidence regarding shared decision-making or patient decision aids in orthopaedics. In the survey, 362 of 639 (57%) randomly selected consultant orthopaedic surgeons responded. Respondents appear representative of consultant orthopaedic surgeons in the UK. Of 272 valid responses, 79% (95% CI, 73-85%) thought patient decision aids a good or excellent idea. There was consensus on the potential helpfulness of patient decision aids and core content. A booklet to take home was the preferred medium/practice model. CONCLUSIONS: Despite the increased emphasis on patient involvement in decision-making, there is little evidence in the medical literature relating to shared decision-making or the use of patient decision aids in orthopaedic surgery. Further research in this area of clinical practice is required. Our survey shows that consultant orthopaedic surgeons in the UK are generally positive about the use of patient decision aids for joint replacement surgery. Survey results could inform future development of patient decision aids for joint replacement practice in the UK. PMID- 18430334 TI - Fast-track assessment clinic: selection of patients for a one-stop hip assessment clinic. AB - INTRODUCTION: The fast-track assessment clinic (FTAC) is a process to select patients who are very likely to require primary total hip replacement. Selected patients can then be seen in a one-off clinic reducing the number of hospital visits, cost to primary care trusts and delay between referral and treatment. PATIENTS AND METHODS: Fifty patients on the waiting list for hip replacement were analysed to see if there were common parameters that led to their inclusion. From these data, fast-track selection criteria (FTSCs) were generated. These FTSCs were used to make a dual comparison of outcomes between 52 patients seen in a traditional clinic. Finally, a pilot study was conducted in which patients fulfilling FTSCs were seen in a designated clinic. RESULTS: An Oxford hip score (OHS) of 34 and above combined with severe loss of joint space, severe marginal osteophytes, or both was common to most patients on the waiting list (84%). FTSCs correctly predicted the outcome of the orthopaedic clinic in 38 patients out of a total of 52. During the pilot stage, positive FTSCs were shown to have a positive predictive value of 92% for joint replacement being carried out and a negative predictive value of 46%. CONCLUSIONS: An OHS of 34 or above combined with complete loss of joint space and/or severe marginal osteophyte formation can be used to select patients who are very likely to need total hip replacement. These patients can be seen in a clinic that combines assessment of surgical indication with medical fitness for surgery. PMID- 18430335 TI - Implementation of and compliance with NICE guidelines in the secondary prevention of osteoporotic fractures in postmenopausal women. AB - Osteoporosis affects more than 1.2 million women in England and Wales. Hip fractures are associated with increased mortality. The total cost of treating osteoporotic fractures in postmenopausal women is expected to rise to pound 2.1 billion by 2010. The National Institute for Health and Clinical Excellence (NICE) has issued guidelines for the secondary prevention of osteoporotic fractures in postmenopausal women. The objective of this study was to review the NICE guidelines and to assess the implementation of and compliance with these guidelines in a district general hospital. PMID- 18430336 TI - Injury to the lateral femoral cutaneous nerve during minimally invasive hip surgery: a cadaver study. AB - INTRODUCTION: A smaller skin incision as part of minimally invasive surgery (MIS) for total hip arthroplasty (THA) is thought to reduce local tissue trauma. Preservation of cutaneous nerves may reduce postoperative pain and improve rehabilitation. The standard lateral approach (SLA) and two MIS approaches (anterolateral [ALA] and posterior [PA]) were compared to determine which incision preserved cutaneous nerves. PATIENTS AND METHODS: Fifteen cadaveric hip specimens were dissected to establish the course of branches of the lateral femoral cutaneous nerve. The number of branches divided by each of the three incisions was recorded. RESULTS: The MIS incisions resulted in significantly less nerve division compared with the SLA. The mean difference between the SLA and the ALA was 4.4 (P < 0.0001), and between the SLA and the PA was 1.4 (P = 0.0005). The ALA divided significantly fewer nerves compared with the PA (mean difference = 3; P = 0.0001). DISCUSSION: There is little evidence of the benefit of one MIS approach over another in the literature. The ALA preserved significantly more cutaneous nerves in this study. These results need corroboration with clinical outcomes to determine their significance. PMID- 18430337 TI - Diagnostic biopsy of lymph nodes of the neck, axilla and groin: rhyme, reason or chance? AB - INTRODUCTION: Improving patient pathways of care is becoming increasingly important in the delivery of timely, appropriate surgical care. With this aim, we analysed the referral and management pathway of patients undergoing diagnostic superficial lymph node biopsy. PATIENTS AND METHODS: A retrospective review of case notes of patients undergoing diagnostic superficial lymph node biopsy over 3 years, 1998-2000 at the Bradford Hospitals NHS Trust. Indication for surgical biopsy was based on clinical suspicion following assessment in the out-patient clinic for the majority, and arrangement of investigations as deemed appropriate. There were no clinical algorithms in use during the study period. RESULTS: There was no evidence for the use of explicit protocols for referral or management. Biopsy was often delayed. Of 268 patients referred from primary care, referral was made to any of 14 hospital Department with 39% (105 of 268) attending more than one outpatient appointment, and 155 (41 of 268) attending more than one department. Eighteen percent (47 of 268) of patients were informed of their diagnosis within 6 weeks of referral and 42% (113 of 268) within 3 months of referral. Nine percent (24 of 268) underwent pre-operative fine needle aspiration cytology. Of patients with enlarged neck nodes, 29% (52/180) had examination of the upper aero-digestive tract. CONCLUSIONS: The study supports the introduction of co-ordinated problem-based referral and management pathways for the management of patients with enlarged superficial lymph nodes supported by regular audits of practice. PMID- 18430338 TI - A significant reduction in paediatric post-tonsillectomy vomiting through audit. AB - INTRODUCTION: Postoperative vomiting occurs more frequently after tonsillectomy than any other commonly performed paediatric operation. Postoperative vomiting is also the commonest cause of morbidity and re-admission following tonsillectomy. We present a successful completed audit cycle and literature review on the subject. PATIENTS AND METHODS: Data on the risk factors for postoperative vomiting, whether the patient vomited and details of the patient's vomitus were collected prospectively on consecutive patients and compared with a gold standard. Changes in practice were agreed and a second cycle performed. RESULTS: Two cycles and a total of 107 patients were included in the audit. A significant reduction in vomiting from 27% to 11% was achieved following the introduction of routine use of intravenous dexamethasone during surgery. CONCLUSIONS: This simple prospective audit of paediatric post-tonsillectomy vomiting has resulted in a statistically significant reduction in vomiting which would appear to be due to use of intra-operative steroids. PMID- 18430339 TI - The ANKLe score: an audit of otolaryngology emergency clinic record keeping. AB - INTRODUCTION: Accurate and legible medical records are essential to good quality patient care. Guidelines from The Royal College of Surgeons of England (RCSE) state the content required to form a complete medical record, but do not address legibility. An audit of otolaryngology emergency clinic record keeping was performed using a new scoring system. PATIENTS AND METHODS: The Adjusted Note Keeping and Legibility (ANKLe) score was developed as an objective and quantitative method to assess both the content and legibility of case notes, incorporating the RCSE guidelines. Twenty consecutive otolaryngology emergency clinic case notes from each of 7 senior house officers were audited against standards for legibility and content using the ANKLe score. A proforma was introduced to improve documentation and handwriting advice was given. A further set of 140 notes (20 notes for each of the 7 doctors) was audited in the same way to provide feedback. RESULTS: The introduction of a proforma and advice on handwriting significantly increased the quality of case note entries in terms of content, legibility and overall ANKLe score. CONCLUSIONS: Accurate note keeping can be improved by the use of a proforma. The legibility of handwriting can be improved using simple advice. The ANKLe score is an objective assessment tool of the overall quality of medical note documentation which can be adapted for use in other specialties. PMID- 18430340 TI - Gastric cancer surgery--a balance of risk and radicality. AB - INTRODUCTION: The aim of this study was to determine whether tailoring the extent of resection would allow radical gastric cancer surgery to be performed safely in a UK population. PATIENTS AND METHODS: A total of 180 consecutive patients (median age 70 years; male:female ratio 2:1) undergoing resection for gastric adenocarcinoma with curative intent were studied. Extent of lymphadenectomy was based upon pre-operative and intra-operative staging, and balanced against the patient's age and fitness. RESULTS: In the study group, 83 patients underwent subtotal or distal partial gastrectomy and 97 patients underwent total or proximal partial gastrectomy. Operative procedures were: D1 lymphadenectomy (n = 62); modified (spleen and pancreas pre-serving) D2 lymphadenectomy (n = 73); D2 lymphadenectomy (n = 42); and extended resection (n = 3). TNM classification was: stage 1 (n = 45); stage 2 (n = 37); stage 3 (n = 61); and stage 4 (n = 37). Of the patients, 48 developed postoperative complications including 17 patients with a major surgical complication. The in-hospital mortality was 1.7% (3 of 180). Predicted mortality according to POSSUM and P-POSSUM was 21.4% and 7.8%, respectively. Disease-specific 5-year survival according to stage was 85.4%, 64.2%, 33.3%, and 6.9%. CONCLUSIONS: By tailoring the extent of resection and balancing risk and radicality, gastric cancer surgery can be performed with low mortality in Western patients. PMID- 18430341 TI - Specialist outreach service for on-table repair of iatrogenic bile duct injuries- a new kind of 'travelling surgeon'. AB - INTRODUCTION: The objective of this study was to determine the outcome of on table repair of iatrogenic bile duct injuries (IBDIs) following laparoscopic cholecystectomy, by specialist hepatobiliary surgeons as an outreach service. PATIENTS AND METHODS: Prospectively collected data on IBDI managed as an outreach service, was analysed retrospectively. The Strasberg classification was used to define types of injury. RESULTS: There were 22 patients. Twenty (91%) had type E 'classical' excision injuries, two had types B and D. Two type E cases had co existent vascular injury both with right hepatic artery injuries; one also had a co-existent portal vein injury. A Roux-en-Y hepaticojejunostomy was used to repair the IBDI in 21 (95%) patients. One type D injury had duct repair over a T tube. No attempt was made to reconstruct the injured hepatic arteries, while the portal vein injury was primarily repaired. The median follow-up was 210 days (range, 47-1088 days). Two patients developed bile leak and cholangitis while another developed transient jaundice. There were no postoperative mortalities. All patients were followed up at our centre. CONCLUSIONS: Repair of IBDI as an outreach service by specialist surgeons is feasible and safe, with minimal disruption to the patient pathway. Prompt recognition and definitive management may help reduce complaints and medicolegal litigation. PMID- 18430342 TI - Living-unrelated donor renal transplantation: an alternative to living-related donor transplantation? AB - INTRODUCTION: An increasing number of living-unrelated, kidney donor transplants are being performed in our unit. We present a comparison of living-unrelated (LURD) and living-related donor (LRD) renal transplant outcomes and analyse influencing factors. PATIENTS AND METHODS: We retrospectively analysed the outcome of all living-donor renal transplants performed at our centre from 1993 to 2004. The parameters studied included patient and graft survival, functioning status of grafts (determined by estimated GFR) at last follow-up and any rejection episodes. Multivariate analysis was performed for recipient and donor age, ethnicity, HLA matching and re-transplants. RESULTS: A total of 322 live donor kidney transplants (LRD, n = 261; LURD, n = 61) were carried out over this period. Mean recipient age was 28 +/- 16 years in the LRD group and 48 +/- 12 years in LURD, while mean age of the donors was 43 +/- 11 years and 48 +/- 10 years, respectively. Caucasians constituted 80% of all the living donors. Amongst LRD, parents were the commonest (58%) donors followed by siblings (35%). In LURD, 80% were spouses. A total of 33 grafts failed, 30 in LRD (11%) and 3 in LURD (5%). Thirteen patients died, 11 (4.2%) in LRD (7 with functioning graft) and 2 (3.3%) in LURD (1 with functioning graft). Acute rejections occurred in 41% recipients in LRD and 35% in LURD (P = 0.37). Estimated GFR was lower in LURD than in LRD (49 +/- 14 versus 59 +/- 29 ml/min/1.73 m(2); P = 0.032). One- and 3 year patient survival for LRD and LURD was 98.7% and 96.3% and 97.7% and 95%, respectively (P = 0.75). One- and 3-year graft survival was equivalent at 94.8% and 92.3% for LRD, and 98.4% and 93.7% for LURD, respectively (P = 0.18). CONCLUSIONS: Outcome of LRD and LURD is comparable in terms of patient and graft survival, acute rejection rate and estimated GFR despite differences in demographics, HLA matching and re-transplants of recipients. PMID- 18430343 TI - Home-made, adapted and modified devices in surgical practice. PMID- 18430348 TI - Children's attitudes toward behavior management techniques used by dentists. AB - PURPOSE: Many behavior management techniques (BMTs) are used in dental offices. The objective of this study was to evaluate how children felt towards the BMT used in the dental office by using the newly invented "attitude meter." METHODS: Two hundred forty children 6 to 17 years old were selected randomly to participate in the study. Each student was asked to watch 8 video scenes of live BMTs. The BMTs used consisted of: (1) tell-show-do (TSD); (2) rewards; (3) general anesthesia; (4) papoose board; (5) hand-holding; (6) mouthprop; (7) voice control; and (8) hand-over-mouth exercise (HOME). After watching each BMT scene, the children were instructed to express their attitude towards the BMT by drawing a "line of favor"--the newly invented attitude meter. RESULTS: It was found that TSD and HOME were the most and least favorite BTM, respectively. Those who had dental experience appeared to have worse attitudes. Older and younger children had different opinions towards some BMTs. The older children preferred the papoose board and hand-holding to the mouthprop. All children preferred the use of the papoose board and hand-holding to voice control. CONCLUSIONS: Children appeared to judge a behavior management technique according to the way it looked. The "line of favor" is a reliable tool to measure attitudes of children over 6 years old. PMID- 18430349 TI - Dental students attitudes regarding online education in pediatric dentistry. AB - PURPOSE: The Atlas of Pediatric Dentistry is a Web-based (Atlas of Pediatric Dentistry Atlas of Pediatric Dentistry (http://depts.washington.edu/peddent/) and disk-deployed comprehensive textbook of pediatric dentistry that features approximately 3,500 frames and 2,500 images, including color clinical photographs, illustrations, and radiographs; There are also end-of-chapter quizzes, laboratory manuals, photo banks, and a scrollable index incorporated as features in the book. The purpose of this study--which is among a series of studies being conducted at the University of Washington--was to evaluate dental student perceptions regarding online/computer-deployed education. METHODS: 55 third year dental students were surveyed regarding their use of the Atlas of Pediatric Dentistry as the textbook resource during a comprehensive, introductory course in pediatric dentistry. The response rate was 84% of 55 dental students. RESULTS: Most students (80%) preferred the online textbook to traditional textbooks. Ninety-six percent rated the educational content of the Atlas as good, very good, or excellent, and all respondents indicated they would recommend the textbook highly to a colleague. Positive perceptions were associated with the: (1) scope of content; (2) large number of images (especially color images); and (3) ease of use. The most negative perceptions were due to technical problems associated with online use of computers running obsolete (Internet) browser software. The majority of respondents (87%) had access to a computer at home. Fifteen percent of the total 46 respondents used printouts of the text material. CONCLUSIONS: Students strongly preferred the online textbook to a traditional text. There continues to be a small number of students who do not have access to a computer at home and whose computers are equipped with obsolete software. To optimize student access to the online program, it is important to plan for students to be able to use computers at the university and to program any online resource so that it is compatible with older browser software. PMID- 18430350 TI - Temperature changes in the deciduous pulp chamber during cavity preparation with the Er:YAG laser. AB - PURPOSE: The purpose of this in vitro study was to evaluate the temperature changes in the pulp chambers of deciduous molars during cavity preparation with an Er:YAG (erbium substituted: yttrium aluminium garnet) laser. METHODS: Two cavities were prepared in 14 maxillary deciduous molars: one with the Er:YAG laser and other with a diamond bur using a high-speed handpiece (control), providing 4 groups with 7 cavities each. The laser treatment was applied for 30 seconds (group 1) and 60 seconds (group 3). The internal pulp temperature was recorded during preparation procedures with a thermocouple, and temperature changes were calculated by the difference between the maximum and initial temperature. RESULTS: There were significant differences (P<.01) between Er:YAG laser irradiation for 60 seconds and 30 seconds and the 2 high-speed drilling groups, which were similar. The Er:YAG laser used for 60 seconds provided the highest temperature alteration (4.01 degrees C). CONCLUSION: Er:YAG laser irradiation increased the temperature of the deciduous teeth compared to the high speed drilling treatment. The temperature increments, however, remained below the critical value (5.5 degrees C). PMID- 18430351 TI - Shear bond strength to primary enamel: influence of Er:YAG laser irradiation distance. AB - PURPOSE: The purpose of this study was to evaluate in vitro the influence of Er:YAG laser irradiation distance on the shear bond strength of an adhesive restorative system to primary enamel. METHODS: Fifty buccal surfaces of extracted human canines were ground and divided into 5 groups (N=10). The control group was etched with 35% phosphoric acid (CA). In the lased groups, the enamel surface treatment was performed with the Er:YAG laser (80mJ/2Hz) by varying the irradiation distance (12, 14, 16, and 17 mm), followed by acid etching. An adhesive agent (Single Bond) was applied on the bonding sites, and resinous cylinders (Filtek Z250) were prepared. Shear bond strength tests were performed in a universal testing machine (0.5 mm/minute). Failure mode was assessed using a X40 magnification stereomicroscope. Data were submitted to statistical analysis by analysis of variance. RESULTS: The means in MPa were: (1) CA=18.76 (+/-6.68); (2) 12 mm=12.73 (+/-5.46); (3) 14 mm=15.9 (+/-6.81); (4) 16 mm=20.1 (+/-6.94); and (5) 17 mm=15.15 (+/-6.81). There was no statistically significant difference (P<.05) among the tested groups. CONCLUSION: The different Er:YAG laser distance irradiations did not influence the adhesive resistance of the resinous system to enamel, even when compared with the control group (acid etching solely). PMID- 18430352 TI - Effect of cleansers and irrigation methods on primary root dentin permeability. AB - PURPOSE: The purpose of this study was to evaluate the effects of cleansers and irrigation methods on the permeability index (PI) in primary root dentin. METHODS: One hundred twelve teeth were extracted and sectioned transversely, discarding their crowns. Then, the roots were distributed according to the cleansers: (1) Dakin (D); (2) Dakin+hydrogen peroxide (DHP); (3) 2% chlorhexidine gel (C); and (4) saline (S). The canals were manually instrumented. Irrigation was performed manually (MI) or ultrasonically (UI). The roots were: (1) made impermeable; (2) filled with 2% methylene blue solution; and (3) longitudinally sectioned. The halves were marked in thirds (cervical, middle, and apical), and the areas of dye penetration were measured. PI data were submitted to factorial analysis of variance and Tukey tests (P<.05). RESULTS: A significant effect of MI/UI, in association with cleansers, was seen on PI. For all thirds, the PI of MI was superior to UI. In the cervical third, MI achieved a high PI when associated with D, DHP, and S. UI associated with S and C produced the highest PI. In the middle third, MI associated with D and DHP produced the highest PI. In the apical third, MI associated with D demonstrated the highest PI. None of the UI and cleanser associations induced differences in PI for the middle and apical thirds. CONCLUSION: MI produced better results than the UI. The D, DHP, and S cleansers demonstrated the highest PIs, suggesting that the combination of MI and these cleansers is recommended for primary teeth to maintain dentin permeability. PMID- 18430353 TI - Colonization of mutans streptococci in Costa Rican children from a high-risk population. AB - PURPOSE: The aim of the study was to determine the colonization of mutans streptococci (MS) in dental biofilm and saliva of children from a high-risk population with low socioeconomic status from San Jose, Costa Rica. METHODS: A total of 68 healthy babies from 8 to 20 months of age (mean age=13 months), with caries-free status and cariogenic feeding habits (eg, nursing bottle- or breast feeding on demand) were examined. Children divided in two groups (<13 months and 14-20 months). Samples from dental biofilm and saliva from each subject were processed and plated in blood agar (BA) and Mitis-Salivarius-Bacitracin agar (MSB) to determine MS percentage. Statistical analysis included the analysis of variance test, and stratification by age of MS levels. RESULTS: Detectable MS levels in dental biofilm and saliva were found in 75% and 72% of children, respectively. Counts higher than 20000 colony forming units (CFU) per ml were found in 4% of saliva and in 35% of biofilm samples. The two groups showed significant differences in MS levels for dental biofilm (P=.036) and saliva (P=.009). Children older than 17 months presented higher MS levels (P<.05). Analysis by MS levels (<0.1%; 0.1-1.0%;>1.0% of MS) The two groups showed an insignificant association with increasing mean age. MS density was associated with mean number of erupted primary molars. CONCLUSIONS: Mutans streptococci colonization of dental biofilm and saliva from high caries risk infants is associated with age and dental development. PMID- 18430354 TI - Occlusal characteristics in cerebral palsy patients. AB - PURPOSE: The purpose of this study was to evaluate the occlusal characteristics of 104 randomly selected, noninstitutionalized patients with a medical diagnosis of cerebral palsy (CP). METHODS: Patients were grouped according to the CP motor disorder and clinical patterns. Occlusal characteristics were collected by one examiner. Nonparametric statistical tests, including Fisher's exact test (P) and chi-square test were used. RESULTS: The results showed that Class II molars were statistically higher (P<.05) for spastic patients. Anterior open bite and an increased overjet were significant (P<.01) in spastic patients with clinical patterns of quadriplegia, double-hemiplegia, and diplegia. Double hemiplegia patients presented a higher incidence of open bite (64%) with an edge-to-edge overjet (45%). A deep overbite was presented in athetoid (50%) and ataxic (67%) patients. A high incidence of tooth wear was observed in the athetoid group (54%). CONCLUSION: Spastic patients presented a high incidence of open bite and Class II malocclusion. A low level of malocclusion was observed in the ataxic group compared to the other groups studied. Further studies would be useful to determine if neurological injuries can be the cause of changes of the craniofacial growth and development patterns resulting in malocclusion in cerebral palsy patients. PMID- 18430355 TI - Apical microleakage of primary teeth root canal filling materials. AB - OBJECTIVE: The objective of this study was to evaluate in vitro apical microleakage of root canal filling materials for primary teeth. METHODS: Fifty extracted primary anterior teeth were distributed into 6 groups: 4 experimental groups; 1 negative control group; and 1 positive control group. Zinc oxide and eugenol (ZOE), Kri paste, calcium hydroxide (Ca(OH)(2)), and Vitapex were inserted into the canal using a lentulo spiral mounted in a slow-speed handpiece. All teeth were then subjected to a dye leakage test using 2% methylene blue as the tracer. RESULTS: The highest dye penetration was recorded for teeth filled with ZOE, with mean values of 9.01+/-1.23 mm. Vitapex showed the lowest dye penetration, with mean values of 4.02+/-1.79 mm. Comparison between groups showed significant difference when the Vitapex and Kri paste groups were compared to the ZOE group (P<.0001). Also, a significant difference between the Vitapex and Ca(OH)2 group (P=.014) was detected. The result showed no difference between Vitapex and Kri paste (P=.658) nor between Kri paste and Ca(OH)(2) (P=.185). CONCLUSIONS: All the resorbable root canal filling materials used in primary teeth showed apical leakage. The highest apical leakage was recorded for ZOE, while Vitapex showed minimum dye leakage. PMID- 18430356 TI - Caries prior to age 3 and breastfeeding: a survey of La Leche League members. AB - PURPOSE: The purpose of this study was to conduct a secondary data analysis of results from a 1985 survey of La Leche League International (LLLI) members to further investigate the relationship between breast-feeding and caries prior to age 3 (CPA3). METHODS: Subjects were 576 LLLI mothers who responded to a 23-item questionnaire concerning the following factors for their oldest child: (1) breast feeding habits; (2) fluoride status; (3) use of antibiotics; (4) carbohydrate ingestion; (5) history of caries; and (6) oral hygiene practices. RESULTS: CPA3 was reported in 10% of all children breast-fed for more than 3 years. Later weaning was found to be significantly associated with CPA3 (odds ratio [OR]=2.03; P=.0001). Late initiation of oral hygiene was marginally associated with CPA3 (OR=0.77; P=.08). Among children who developed CPA3, bivariate analyses found a greater frequency of breast-feeding (P=.012) and presence of night-time breast feeding (P=.049) to be associated with caries detected at an earlier age. Children with caries on their maxillary incisors were more likely to have been breast-fed at night (P=.027) and more frequently during the night (P=.032). CONCLUSION: This retrospective study, based on a report of La Leche League International members, found later weaning to be significantly associated with an increased likelihood of developing CPA3. PMID- 18430357 TI - Perceptions of Hispanic American schoolchildren to the risk factors for oral cancer. AB - PURPOSE: The purpose of this study was to assess the knowledge of high schoolchildren on oral cancer risk factors in southern Texas. METHODS: A 39-item, self-administered questionnaire was completed by 1,667 students (14-18 years) in 4 high schools chosen by stratified cluster sampling. RESULTS: The effective response rate was 67%. Ninety-five percent of the students were Hispanic American, 55% had not heard about oral cancer, and 60% failed to identify the 2 most common early signs of oral cancer. CONCLUSIONS: Lack of awareness of oral cancer risk factors indicate the need for improved health education directed towards minority population, especially in underprivileged areas. PMID- 18430358 TI - Orofacial findings and dental treatment in an 8-year-old patient with trisomy 18: a case report. AB - Trisomy 18 is characterized by: psychomotor disabilities, dysmorphic features, organ malformations, including mental retardation, growth deficiency, poor motor ability, micrognathia, microcephaly, congenital heart defects, and kidney abnormalities. The oral findings typically observed in these patients are: cleft lip and a high, narrow, and sometimes cleft palate. The degree of severity of the malformations is directly related to life expectancy. Only 5% to 10% of affected infants survive beyond the first year of life. Although trisomy 18 has been widely investigated from a medical standpoint, there is a lack of reports addressing the oral manifestations and dental treatment approach in affected children, presumably due to their shortened life expectancy. The purpose of this article was to present the case of an 8-year-old child diagnosed with trisomy 18 and address the clinical features observed--emphasizing the disease-specific oral, craniofacial, and dental findings. Dental care management of the patient is described. PMID- 18430359 TI - Endodontic treatment of a multirooted permanent maxillary canine. AB - The purpose of this paper was to report an unusual case of multirooted permanent maxillary canine. A 16-year-old female patient presented with pain and swelling related to the upper right permanent canine. Radiographic examination revealed a multirooted permanent maxillary canine--an unusual finding. Endodontic treatment was performed after amputation of 2 extra roots, and then the tooth was intentionally reimplanted. The prevalence of birooted permanent mandibular canines in the Japanese population has been reported, but the prevalence of this 3-rooted maxillary canine is still unknown. This report also states the potential etiological factors, effects on the developing dentition, and various treatment options for the multirooted maxillary permanent canine. PMID- 18430360 TI - 5-HT1A/7 receptor agonist excites cardiac vagal neurons via inhibition of both GABAergic and glycinergic inputs. AB - AIM: To study the synaptic mechanisms involved in the 5-hydroxytryptamine1A/7 (5 HT1A/7) receptor-mediated reflex control of cardiac vagal preganglionic neurons (CVPN). METHODS: CVPN were retrogradely labeled and identified in brain stem slices of newborn rats, and their synaptic activity was examined using whole-cell patch-clamp. RESULTS: 8-Hydroxy-2-(di-N-propylamino) tetralin (8-OH-DPAT), an agonist of 5-HT1A/7 receptors, had no effect on the glutamatergic inputs of CVPN. In contrast, it significantly decreased the frequency and the amplitude of both the GABAergic and the glycinergic spontaneous inhibitory postsynaptic currents (sIPSC). 8-OH-DPAT also caused significant amplitude decrease of the GABAergic currents evoked by stimulation of the nucleus tractus solitarius. Both the frequency inhibition and the amplitude inhibition of the GABAergic and the glycinergic sIPSC by 8-OH-DPAT had dose-dependent tendencies and could be reversed by WAY-100635, an antagonist of 5-HT1A/7 receptors. In the pre-existence of tetrodotoxin, 8-OH-DPAT had no effect on the GABAergic or the glycinergic miniature inhibitory postsynaptic currents, and had no effect on the GABAergic or the glycinergic currents evoked by exogenous GABA or glycine. CONCLUSION: The 5 HT1A/7 receptor agonist excites CVPN indirectly via the inhibition of both the GABAergic and glycinergic inputs. These findings have at least in part revealed the synaptic mechanisms involved in the 5-HT1A/7 receptor-mediated reflex control of cardiac vagal nerves in intact animals. PMID- 18430361 TI - Bilobalide inhibits 6-OHDA-induced activation of NF-kappaB and loss of dopaminergic neurons in rat substantia nigra. AB - AIM: To investigate the effects of bilobalide on the activation of NF-kappaB, and apoptosis of dopaminergic neurons induced by 6-hydroxydopamine (6-OHDA). METHODS: A rat model of Parkinson's disease was produced with a unilateral infusion of 6 OHDA (8 mug) into the substantia nigra par compact. Bilobalide was administered 5, 10, and 20 mg/kg (ip) once a day for 7 d, starting 6 d prior to the 6- OHDA infusion. The rats were subjected to locomotor activity and rotational behavior testing 2 or 3 weeks after the 6-OHDA infusion. The expressions of tyrosine hydroxylase (TH) and NF-kappaB p65 were examined by immunofluorescence. The loss of dopaminergic neurons was detected by Nissl's staining. Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling was used to identify apoptosis. RESULTS: The behavioral changes due to 6-OHDA were significantly restored by bilobalide pretreatment. Bilobalide inhibited the 6-OHDA-induced loss of TH-positive neurons, decreased the activation of NF-kappaB, and protected dopaminergic neurons from apoptosis remarkably. CONCLUSION: NF-kappaB activation contributes to the 6-OHDA-induced loss of dopaminergic neurons, and the inhibition of the NF-kappaB pathway is likely to be involved in the neuroprotective effect of bilobalide. PMID- 18430362 TI - Biological characters of [18F]O-FEt-PIB in a rat model of Alzheimer's disease using micro-PET imaging. AB - AIM: To evaluate whether the newly-synthesized positron emission tomography (PET) tracer, [18F]2-(4'-(methylamino)phenyl)-6-fluoroethoxy- benzothiazole ([18F] O FEt-PIB), could bind to beta-amyloid aggregates in a rat model of Alzheimer's disease (AD) using micro-PET. METHOD: [18F]O-FEt-PIB was synthesized and purified by radio HPLC. PET imaging was performed with a R4 rodent model scanner in 3 model and 3 control rats. Dynamic PET scans were performed for 40 min in each rat following an injection of approximately 37 MBq of [18F]O-FEt-PIB. Static scans were also performed for 15 min in each rat. PET data were reconstructed by a maximum posteriori probability algorithm. On the coronal PET images, regions of interest were respectively placed on the cortex, hemicerebrum [including the hippocampus and thalamus (HT)], and were guided by a 3-D digital map of the rat brain or the brain images of [18F]2-Deoxy-2-fluoro-D-glucose ([18F]FDG) in normal rats. Time-activity curves (TAC) were obtained for the cerebrum and cerebellum. The activity difference value (ADV) between 2 hemicerebrums was also calculated. RESULTS: The TAC for [18F]O-FEt-PIB in the cerebrum or cerebellum peaked early (at approximately 2 min), but washed out a little slowly. In the dynamic and static micro-PET images, increased radioactivity was found in the area of the right HT in the model rats where infused with beta-amyloid (1-40). No distinct difference of radioactivity was found between the right and left HT areas in the control rats. The ADV(HT) was approximately 14.6% in the AD model rats and approximately 4 times greater than that of the control rats (3.9%). CONCLUSION: To our knowledge, this study is the first to evaluate a small molecular PET probe for the beta-amyloid deposits in vivo using micro-PET imaging in an AD-injected rat model. The suitable biological characters showed that the tracer had potential to be developed as a probe for detecting beta-amyloid plaques in AD. PMID- 18430363 TI - Curcumin inhibits cellular cholesterol accumulation by regulating SREBP 1/caveolin-1 signaling pathway in vascular smooth muscle cells. AB - AIM: To investigate the protective effect and the possible mechanism of curcumin on anti-atherosclerosis. METHODS: Morphological changes of atherosclerotic lesions taken from apoE knockout (apoE-/-) mice were determined by hematoxylin- eosin staining. Intracellular lipid droplets and lipid levels were assayed by oil red O staining and HPLC. The protein expression of caveolin-1 was quantified by Western blotting. Translocation and the expression of sterol response element binding protein-1 (SREBP-1) were indirectly detected by an immunofluorescence analysis. RESULTS: The administration of 20 mg. kg(-1 ). d(-1 )curcumin to apoE-/ mice for 4 months induced a 50% reduction of atherosclerotic lesions and yielded a 5- fold increase in the caveolin-1 expression level as compared to the model group. Rat vascular smooth muscle cells (VSMC) pretreated with 50 mg/L ox-lipid density lipoprotein(ox-LDL) for 48 h increased cellular lipid contents, and stimulated SREBP-1 translocation, but decreased the caveolin-1 expression level. Lipid-loaded cells exposed to curcumin at various concentrations (12.5, 25, and 50 micromol/L) for different durations (0, 6, 12, 24, and 48 h) significantly diminished the number and area of cellular lipid droplets, total cholesterol, cholesterol ester, and free cholesterol accompanying the elevation of the caveolin-1 expression level (approximately 3-fold); the translocation of SREBP-1 from the cytoplasm to the nucleus was inhibited compared with the models. Lipid loaded VSMC exposed to N-acetyl- Leu-Leu-norleucinal, a SREBP-1 protease inhibitor, showed increased nuclear translocation of SREBP-1, reduced caveolin-1 expression level, and upregulated cellular lipid levels. CONCLUSION: Curcumin inhibits ox-LDL-induced cholesterol accumulation in cultured VSMC through increasing the caveolin-1 expression via the inhibition of nuclear translocation of SREBP-1. PMID- 18430364 TI - Functional subtypes of renal alpha1-adrenoceptor in diabetic and non-diabetic 2K1C Goldblatt renovascular hypertension. AB - AIM: This study investigates the subtypes of the alpha1-adrenoceptor mediating the adrenergically-induced renal vasoconstrictor responses in streptozotocin induced diabetic and non-diabetic 2-kidney one clip (2K1C) Goldblatt hypertensive rats. METHODS: The renal blood flow responses to renal nerve stimulation, noradrenaline, phenylephrine, and methoxamine were measured in the absence and presence of nitrendipine, 5-methylurapidil, chloroethylclonidine and BMY 7378. RESULTS: The renal vasoconstrictor responses were markedly attenuated by nitrendipine and 5- methylurapidil in the diabetic rats (all P< 0.05). In the non diabetic rats, these responses were markedly attenuated by nitrendipine, 5 methylurapidil, and BMY 7378 (all P< 0.05). In both experimental groups, chloroethylclonidine markedly accentuated the renal vasoconstrictions caused by all the adrenergic stimuli (all P< 0.05). CONCLUSION: These observations indicate that alpha 1A-adrenoceptor subtypes play a major role in mediating adrenergically induced renal vasoconstriction in the diabetic 2K1C Goldblatt hypertensive rats. In the non-diabetic 2K1C Goldblatt hypertensive rats, contributions of alpha 1A and alpha 1D-adrenoceptor subtypes were proposed. Apart from post-synaptic alpha 1-adrenoceptors, both in the diabetic and non-diabetic 2K1C Goldblatt hypertensive rats, the potential involvement of presynaptic alpha 1- adrenoceptors is also suggested. PMID- 18430365 TI - Increased secretion and expression of amylin in spontaneously diabetic Goto Kakizaki rats treated with rhGLP-1 (7-36). AB - AIM: To investigate the effect of recombined human glucagon-like peptide 1 (rhGLP 1 [7-36]) on the secretion and expression of amylin in Goto-Kakizaki (GK) rats. METHODS: The GK rats were treated with rhGLP-1 (7-36) 56 and 133 mug/kg subcutaneously for 12 weeks. The fasting and post-prandial blood glucose levels were measured. The plasma amylin concentration was measured by ELISA. The transcription levels of amylin and insulin mRNA were evaluated by fluorescent quantitative- PCR. Immunohistochemistry was utilized to detect the amylin protein. Histological examination was assayed by light microscopy. RESULTS: Treatment with rhGLP-1 (7-36) caused a significant reduction of post-prandial blood glucose levels in the GK rats (P<0.05). The plasma amylin levels of the GK rats were lower than those of Wistar rats after the glucose administration (P<0.01). Treatment with rhGLP-1 (7-36) exhibited a marked elevation of the glucose-stimulated plasma amylin level (P<0.05) and slight histological amelioration. The amylin expression was augmented in the rhGLP-1 (7-36)-treated GK rat pancreas. Amylin and insulin mRNA were also highly expressed in the treated GK rats (P<0.05). However, the ratio of amylin to insulin mRNA was significantly decreased by treatment with rhGLP-1 (7-36). CONCLUSION: RhGLP-1 (7 36) stimulates the secretion and expression of amylin, and exerts a beneficial effect on the ratio of amylin to insulin mRNA. These findings suggest that GLP-1 and GLP-1 analogs are ideal candidates for the treatment of type 2 diabetes. PMID- 18430366 TI - Nitroglycerin enhances proliferation and osteoblastic differentiation in human mesenchymal stem cells via nitric oxide pathway. AB - AIM: To investigate the effect of nitroglycerin (NTG) on cell proliferation and osteoblastic differentiation of human bone marrow-derived mesenchymal stem cells (HBMSC) and its mechanisms. METHODS: Primary HBMSC were cultured in osteogenic differentiation medium consisting of phenol red-free alpha-minimum essential media plus 10% fetal bovine serum (dextran-coated charcoal stripped) supplemented with 10 nmol/L dexamethasone, 50 mg/L ascorbic acid, and 10 mmol/L beta glycerophosphate for inducing osteoblastic differentiation. The cells were treated with NTG (0.1-10 micromol/L) alone or concurrent incubation with different nitric oxide synthase (NOS) inhibitors. Nitric oxide (NO) production was measured by using a commercial NO kit. Cell proliferation was measured by 5 bromodeoxyuridine (BrdU) incorporation. The osteoblastic differentiation of HBMSC culture was evaluated by measuring cellular alkaline phosphatase (ALP) activity and calcium deposition, as well as osteoblastic markers by real-time RT-PCR. RESULTS: The treatment of HBMSC with NTG (0.1-10 micromol/L) led to a dose dependent increase of NO production in the conditional medium. The release of NO by NTG resulted in increased cell proliferation and osteoblastic differentiation of HBMSC, as evidenced by the increment of the BrdU incorporation, the induction of ALP activity in the early stage, and the calcium deposition in the latter stage. The increment of NO production was also correlated with the upregulation of osteoblastic markers in HBMSC cultures. However, the stimulatory effect of NTG (10 micromol/L) could not be abolished by either N(G ) -nitro-L-arginine methyl ester, an antagonist of endothelial NOS, or 1400W, a selective blocker of inducible NOS activity. CONCLUSION: NTG stimulates cell proliferation and osteoblastic differentiation of HBMSC through a direct release of NO, which is independent on intracellular NOS activity. PMID- 18430367 TI - Downregulation of STEAP4, a highly-expressed TNF-alpha-inducible gene in adipose tissue, is associated with obesity in humans. AB - AIM: To determine the relationship between six-transmembrane epithelial antigen of the prostate 4 (STEAP4) expression and obesity. METHODS: RT-PCR and immunoblot analyses were performed to determine the differential expressions of STEAP4 mRNA and protein, respectively, in human omental adipose tissue from obese patients and normal weight controls. The expression pattern of STEAP4 mRNA in various human tissues was determined by RT-PCR. The subcellular localization of the STEAP4 protein in human adipose tissue was confirmed by immunohistochemistry. Finally, we confirmed that cultured human omental adipose tissue undergoes TNF alpha-mediated regulation of the STEAP4 expression. RESULTS: STEAP4 mRNA and protein levels were downregulated in omental adipose tissue from obese patients relative to normal controls. The STEAP4 expression was most abundant in human adipose tissue. An immunohistochemical analysis confirmed that STEAP4 was associated with the plasma membrane of adipocytes. The STEAP4 expression was induced by TNF-alpha in a dose-dependent manner in human adipose tissue. CONCLUSION: STEAP4 was abundantly expressed in human adipose tissue, and the STEAP4 expression was significantly downregulated in obese patients. STEAP4 localized to the plasma membrane of adipocytes, and the STEAP4 expression was induced by TNF-alpha in adipose tissue. These data suggest that STEAP4 may play a significant role in the development of human obesity. PMID- 18430368 TI - Chronic activation of neutral ceramidase protects beta-cells against cytokine induced apoptosis. AB - AIM: To investigate the activity and expression of neutral ceramidase (N-CDase) in the insulin-secreting cell line INS-1 and its role in the cellular response to cytokines. METHODS: HPLC, Western blotting, and quantitative real-time PCR were performed to detect the activity and expression of N-CDase in INS-1 cells treated with a cytokine mixture (5 ng/mL interleukin-1beta, 10 ng/mL TNF-alpha, and 50 ng/mL interferon-gamma). The expression and activity of N-CDase in the INS-1 cells were specifically inhibited using N-CDase-siRNA transfection. Annexin V fluorescein- isothiocyanate/propidium iodide flow cytometry was used to assess apoptosis in the INS-1 cells. RESULTS: The INS-1 cells exhibited some basal N CDase activity, and cytokines induced a time-dependent delay in the activation of NCDase. As a result, the activation of N-CDase was first detectable at 8 h after stimulation. It peaked at 16 h and remained elevated at 24 h. Cytokines also upregulated the mRNA and protein expression of N-CDase in the INS-1 cells. Furthermore, when N-CDase activity was inhibited by RNA interference, cytokine induced apoptosis in the INS-1 cells was markedly increased. CONCLUSION: The N CDase pathway is active in INS-1 cells, and the chronic activation of N-CDase is involved in the pathological response of beta-cells to cytokines, potentially providing protection against cytokine toxicity. PMID- 18430369 TI - Effect of long-term partial bladder outlet obstruction on caldesmon isoforms and their correlation with contractile function. AB - AIM: In the present study, we investigate the expression of caldesmon (CAD) isoforms in rabbit detrusor smooth muscles (DSM) during the progression of partial bladder outlet obstruction and relate them with the time course of obstruction. METHODS: Detrusor samples were obtained from the bladders of rabbits with partial bladder outlet obstruction and sham-operated control rabbits after 1, 2, 4, and 8 weeks of obstruction. Contractile responses to field stimulation and carbachol were determined in the isolated bladder strips. Western blotting was used to determine the relative levels of CaD isoform expression at the protein levels. RESULTS: The contractile responses decreased progressively over the course of obstruction. The expression of l-CaD increased significantly to approximately the same extent as the 1-4-week obstructed groups and further in the 8-week obstructed group. The expression of h-CaD increased in all of the obstructed bladders, but at significantly higher levels in the 1-2-week obstructed bladders compared to the control and 4-8-week obstructed bladders. CONCLUSIONS: The changes in the isoforms of CaD may be part of the molecular mechanism for bladder compensation following partial bladder outlet obstruction. The overexpression of l-CaD and the h-CaD/l-CaD ratio could be markers for the status of DSM remodeling and dysfunction. PMID- 18430370 TI - Adenovirus-mediated expression of SSAT inhibits colorectal cancer cell growth in vitro. AB - AIM: To construct a recombinant adenovirus that can express human spermidine/ spermine N1-acetyltransferase (SSAT) and detect its inhibitory effect on colorectal cancer cell growth in vitro. METHODS: A 516 bp cDNA of SSAT was amplified and cloned into a pGL3-hTERT plasmid. The pGL3-hTERT-SSAT recombinant was digested, and the small fragment was cloned into the shuttle vector pAdTrack. The pAdTrack-hTERT-SSAT plasmids were recombined with pAdEasy-1 vectors in AdEasy 1 cells. Positive clones were selected and transfected into the HEK293 packaging cells (transformed human embryonic kidney cells) after they were linearized by PacI. The process of adenovirus packaging and amplification was monitored by green fluorescent protein (GFP) expression. The SSAT protein levels were determined by Western blotting, and the intracellular polyamine content was detected by reverse-phase high performance liquid chromatography. The MTS (3-(4, 5-dimethylthiaol-2-yl)-5-(3-carboxy-methoxyphenyl)-2-(-4- sulfophenyl)-2H tetrazolium, inner salt) and colony-forming assays were used to analyze the gene transduction efficiency and effect on the growth of HT-29 and LoVo cells. A viable cell count was used to determine the cell growth with or without exogenous polyamines. RESULTS: The GFP expression in 293 cells during virus packing and amplification was observed by fluorescence microscopy. Western blotting results demonstrated that Ad-hTERT-SSAT could increase the expression of SSAT, and consequently, spermidine and spermine were reduced to low levels. The MTS and colony-forming assay results showed that HT-29 and LoVo cell growth were significantly inhibited, and the inhibitory effect could be partially reversed by exogenous spermidine and spermine. CONCLUSION: The successfully constructed recombinant adenovirus Ad-hTERT-SSAT could accelerate polyamine catabolism and inhibit the colorectal cell growth in vitro. It also has therapeutic potential in the treatment of colorectal cancer. PMID- 18430371 TI - Reversal effect of Ganoderma lucidum polysaccharide on multidrug resistance in K562/ADM cell line. AB - AIM: To investigate the reversal effect of Ganoderma lucidum polysaccharides (Gl PS) on multidrug resistance (MDR) in the adriamycin (ADM)-resistant leukemic cell line K562/ADM. METHODS: Cytotoxicity was assayed by 3-(4,5-dimethylthiazol- 2-yl) 2,5-diphenyltetrazolium bromide method; the ADM concentration in cells was determined by flow cytometry and confocal laser scanning microscopy techniques; the expression of P-glycoprotein was assayed by flow cytometry; and the mRNA expression levels of MDR-1 and MDR-associated protein (MRP)1 were determined by RT-PCR. RESULTS: Gl-PS reversed MDR in K562/ADM cells. Gl-PS obviously reversed the resistance of K562/ADM to doxorubicin. The reversing factors of Gl-PS at 10 and 20 mg/L were 6.46 and 6.80, respectively. MDR-1 and MRP1 transcription were downregulated by 10 and 50 mg/L Gl-PS. CONCLUSION: Gl-PS can reverse the MDR by downregulating the expression of MDR-1 and MRP1 in K562/ADM cells. PMID- 18430372 TI - Berberine inhibits cyclin D1 expression via suppressed binding of AP-1 transcription factors to CCND1 AP-1 motif. AB - AIM: To verify the suppressive effect of berberine on the proliferation of the human pulmonary giant cell carcinoma cell line PG and to demonstrate the mechanisms behind the antitumoral effects of berberine. METHODS: The proliferative effects of PG cells were detected by 3-(4,5-dimethylthiazol-2-yl) 2,5- diphenyltetrazolium bromide colorimetry. The cell cycle was examined by flow cytometry. The expression level of cyclin D1 was detected by RT-PCR. The activities of the activating protein-1 (AP-1) and NF-kappaB signaling pathways related to cyclin D1 were examined by luciferase assay. The cytoplasmic level of c-Jun was detected by Western blot analysis. An electrophoretic mobility shift assay was used to examine the binding of transcription factors to the cyclin D1 gene (CCND1) AP-1 motif. RESULTS: The results showed that the proliferation of PG cells treated with different concentrations (10, 20, and 40 microg/mL) of berberine for 24 and 48 h was suppressed significantly compared to the control group. After treatment with berberine, the proportion of PG cells at the G0/G1 phase increased, while cells at the S and G2/M phases decreased. Berberine could inhibit the expression of cyclin D1 in PG cells. Berberine inhibited the activity of the AP-1 signaling pathway, but had no significant effect on the NF-kappaB signaling pathway. Berberine suppressed the expression of c-Jun and decreased the binding of transcription factors to the CCND1 AP-1 motif. CONCLUSION: Berberine suppresses the activity of the AP-1 signaling pathway and decreases the binding of transcription factors to the CCND1 AP-1 motif. This is one of the important mechanisms behind the antitumoral effects of berberine as a regulator of cyclin D1. PMID- 18430373 TI - Identification of antiviral mimetic peptides with interferon alpha-2b-like activity from a random peptide library using a novel functional biopanning method. AB - AIM: To screen for interferon (IFN) alpha-2b mimetic peptides with antiviral activity. METHODS: Selecting IFN receptor-binding peptides from a phage-display heptapeptide library using a novel functional biopanning method. This method was developed to identify peptides with activity against vesicular stomatitis virus (VSV) inducing cytopathic effects on WISH cells. RESULTS: Sixteen positive clones were obtained after 3 rounds of functional selection. Ten clones were picked from these positive clones according to the results of phage ELISA and were sequenced. The amino acid sequences homologous to IFN alpha-2b were defined by residues AB loop 31-37, BC loop 68-74, C helix 93-99, CD loop 106-112, D helix 115-121, DE loop 132-138, and E helix 143-161. Two of the peptides, designated clones T3 and T9, aligned with the IFNAR2-binding domains (AB loop and E helix), were synthesized and designated as IR-7 and KP-7, respectively. Both KP-7 and IR-7 were found to compete with GFP/IFN alpha-2b for receptor binding and mimicked the antiviral activity of IFN alpha -2b cooperatively. CONCLUSION: Two IFN alpha-2b mimetic peptides with antiviral activity were derived from a phage-display heptapeptide library using a novel functional selection method. PMID- 18430375 TI - Uterine asymmetry and dysmenorrhea in a young woman. PMID- 18430374 TI - Synergy of gemcitabine and lidamycin associated with NF-kappaB downregulation in pancreatic carcinoma cells. AB - AIM: To investigate the effects on human pancreatic cancer PANC-1 and SW1990 cells using a combination of lidamycin (LDM) and gemcitabine. METHODS: A 3- (4,5 dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was used to determine the growth inhibition of drugs in PANC-1 and SW1990 cells. The effects on apoptosis were measured by terminal uridine deoxynucleotidyl transferase dUTP nick end labeling assay and flow cytometry combined with fluorescein- isothiocyanate-Annexin V/propidium iodide staining. The activity of caspase-3 was measured with a special assay kit. The mitochondrial membrane potential was determined by confocal microscopy analyses. The level of mRNA encoding K-ras in the cells was determined by RT-PCR analysis. The expression of K-ras, NF-kappaB, and Bcl-2 was detected by Western blotting analysis. RESULTS: There was a significant reduction in proliferation in the pancreatic cancer cell lines treated with a combination of gemcitabine and LDM. The overall growth inhibition directly correlated with apoptotic cell death. LDM potentiated the gemcitabine induced cell killing by reducing mitochondrial membrane potential and increasing the caspase-3 activity. Notably, the K-ras mRNA level was significantly reduced with the combination of gemcitabine and LDM. The results for K-ras, NF-kappaB, and Bcl-2 proteins also showed downregulation in the combination group relative to the single-agent treatment and the untreated control. CONCLUSION: LDM can potentiate the growth inhibition induced by gemcitabine in human pancreatic cancer cells, and the synergy may be associated with NF-kappaB downregulation. PMID- 18430376 TI - Oral contraception without prescription? Help yourself. PMID- 18430377 TI - Sonographic measurement of the lower uterine segment thickness: is it truly predictive of uterine rupture? PMID- 18430378 TI - Vaginal hysterectomy: dispelling the myths. PMID- 18430380 TI - Fetal heart rate response to maternal hypocapnia and hypercapnia in late gestation. AB - OBJECTIVE: To examine the effects of acute maternal hypocapnia and hypercapnia on electronic fetal heart rate (FHR) patterns in late gestation. METHODS: Thirty five women with healthy singleton pregnancies performed a modified carbon dioxide (CO2) rebreathing procedure between 34 and 38 weeks of pregnancy. Prior to rebreathing, subjects hyperventilated for five minutes to reduce end-tidal CO2 tensions (PETco2) below 23 Torr (hypocapnia). During rebreathing, PETco2 progressively increased from hypocapnia to hypercapnia (PETco2 = 40-60 Torr) at a constant hyperoxic end-tidal O2 tension of 150 Torr. FHR responses were classified using standardized guidelines over four periods: 20 minutes before rebreathing (pretest), during hypocapnia and hypercapnia, and 20 minutes after rebreathing (post-test). RESULTS: Mean baseline FHR measures (SD) over the four test periods were 138(8), 144(10), 132(11), and 137(9) beats per minute (bpm). All pairwise comparisons were statistically significant except the pretest versus post-test comparison (P < 0.05, Tukey-Kramer multiple comparisons test). A single tachycardia episode of 170 bpm was recorded in the post-test period. In 20 subjects FHR variability changed from moderate in the pretest period to minimal during hypocapnia and/or hypercapnia. All but two returned to moderate FHR variability in the post-test period. One other fetus with minimal post-test variability had moderate values in the three preceding test periods. CONCLUSION: Electronic FHR parameters remained within normal limits for third-trimester fetuses with the exception of one fetus that experienced tachycardia. Acute maternal hypocapnia and hypercapnia over the range studied had no adverse effects on fetal well-being. These results support the safety of the modified CO2 rebreathing procedure for research in healthy, low-risk pregnancy. PMID- 18430381 TI - Bed rest and activity restriction for women at risk for preterm birth: a survey of Canadian prenatal care providers. AB - OBJECTIVE: To explore the practices of Canadian obstetricians, family physicians, and midwives in recommending bed rest or activity restriction for women at risk for preterm birth (PTB) and to assess the decisional conflict experienced by care providers when they recommend these therapies. METHODS: A self-administered mail survey of prenatal care providers was carried out using Dillman's Tailored Design Method. Analysis included descriptive statistics and analysis of variance. RESULTS: The survey was distributed to 1441 potential participants; of these, 1172 were eligible participants, and 516 (44.2%) completed the survey. For women at risk of PTB, 60 of 170 obstetricians (35%), 88 of 206 family practitioners (42.7%), and 30 of 140 midwives (21.4%) recommended bed rest in hospital; 110 of 170 obstetricians (64.7%), 144 of 206 family practitioners (69.9%), and 73 of 140 midwives (52.1%) recommended bed rest at home. These recommendations occurred despite the response from about two thirds of each professional group that the effectiveness of bed rest was in the fair-to-poor range in helping to prevent PTB. The mean score on the Provider Decision Process Assessment Instrument, measuring decisional conflict for all care provider groups, was 30 (SD 7.4) (possible score range 12-60). There were no significant differences in decisional conflict scores among provider groups (F [2,347] = 2.24; P = 0.11). CONCLUSION: Care providers have been discouraged from routinely recommending bed rest for women at risk of PTB because of potential adverse side effects. This study demonstrates that most Canadian prenatal care providers have not been persuaded to incorporate these recommendations into practice. Except for women with multiple gestation, there is inconsistent practice in recommending bed rest and activity restriction. Additionally, Canadian prenatal care providers have some decisional conflict about using this therapy. These results provide some of the first Canadian perspectives on the practice of prescribing therapeutic bed rest for PTB. PMID- 18430382 TI - Access to abortion: what women want from abortion services. AB - OBJECTIVE: Whether Canadian physicians can refuse to refer women for abortion and whether private clinics can charge for abortions are matters of controversy. We sought to identify barriers to access for women seeking therapeutic abortion and to have them identify what they considered to be most important about access to abortion services. METHODS: Women presenting for abortion over a two-month period at two free-standing abortion clinics, one publicly funded and the other private, were invited to participate in the study. Phase I of the study involved administration of a questionnaire seeking information about demographics, perceived barriers to access to abortion, and what the women wanted from abortion services. Phase II involved semi-structured interviews of a convenience sample of women to record their responses to questions about access. Responses from Phase I questionnaires were compared between the two clinics, and qualitative analysis was performed on the interview responses. RESULTS: Of 423 eligible women, 402 completed questionnaires, and of 45 women approached, 39 completed interviews satisfactorily. Women received information about abortion services from their physicians (60.0%), the Internet (14.8%), a telephone directory (7.8%), friends or family (5.3%), or other sources (12.3%). Many had negative experiences in gaining access. The most important issue regarding access was the long wait time; the second most important issue was difficulty in making appointments. In the private clinic, 85% of the women said they were willing to pay for shorter wait times, compared with 43.5% in the public clinic. CONCLUSION: Physicians who failed to refer patients for abortion or provide information about obtaining an abortion caused distress and impeded access for a significant minority of women requesting an abortion. Management of abortion services should be prioritized to reflect what women want: particularly decreased wait times for abortion and greater ease and convenience in booking appointments. Since many women are willing to pay for services in order to have an abortion within one week, this option should be considered by policy makers. PMID- 18430384 TI - Two cases of umbilical vein thrombosis, one with associated portal vein thrombosis. AB - BACKGROUND: Thrombosis of the umbilical vessels is extremely rare. It is unpredictable and is associated with high perinatal mortality. Few cases with neonatal survival have been reported. CASES: We identified two instances of umbilical vein thrombosis occurring two weeks apart in the same unit. Each case was identified at the time of delivery and each had a good outcome. In the first case, a 22-year-old primigravid woman had severe fetal bradycardia develop in the second stage of labour. Delivery was expedited with vacuum extraction, and a large thrombus in the umbilical vein was noted. In the second case, a 23-year-old multigravid woman underwent emergency Caesarean section because of a non reassuring fetal heart tracing before the onset of labour. An umbilical cord thrombus was noted at delivery. The infant was later found to have a portal vein thrombus but subsequently did well. CONCLUSION: When a neonate is found to have a thrombosis in the umbilical cord, it is prudent to rule out portal vein thrombosis. End-organ damage in the infant should be considered, and a thrombophilia screen is warranted. PMID- 18430383 TI - Barriers and motivations for women to participate in cardiovascular trials. AB - BACKGROUND: Cardiovascular disease is one of the major causes of mortality in Canadian women. However, the number of women recruited into cardiovascular clinical trials continues to be low relative to the prevalence of cardiovascular disease. To determine why the recruitment of women into cardiovascular trials continues to be problematic, we prospectively examined the barriers and motivations of women contacted for the Raloxifene Use for The Heart (RUTH) study. METHODS: Two hundred seventy postmenopausal women were surveyed. Regardless of study eligibility, women were asked to comment on their reasons for participating or not participating in cardiovascular trials in an open-ended single question format. All answers were recorded and grouped accordingly. RESULTS: Fifty-four percent of women surveyed indicated they would not participate in clinical research, and 46% indicated that they would. Motivations reported for participating in a cardiovascular clinical trial included personal health benefits (82.2%), interest in research (44.1%), and the possibility of benefiting society (29.1%). Reasons for declining participation included personal illness (24.8%), transportation issues (17.9%), reluctance to increase medication (15.2%), and concern about adverse health effects (13.1%). CONCLUSION: As reported by our cohort of postmenopausal women, the major barrier to entering a randomized controlled trial related to cardiovascular disease was the perception that participation would present an increased burden on health and time. Our experience suggests that researchers need to educate women on the importance of clinical trials and that they also need to provide practical solutions to barriers such as transportation. PMID- 18430385 TI - Septic uterus after uterine artery embolization for uterine myomas triggered by endometrial biopsy. AB - BACKGROUND: Women who undergo uterine artery embolization (UAE) and subsequently have heavy vaginal bleeding require assessment to establish the cause. Endometrial sampling in such women should not necessarily carry more than the usual risk. CASES: Two women who had undergone UAE presented with recurrence of heavy vaginal bleeding. In order to rule out possible endometrial malignancy, we performed an endometrial biopsy. Both patients had large and necrotic intramural myomas adjacent to the endometrium. They developed septic uterus shortly after endometrial biopsy and each required a hysterectomy. The postoperative course in the first case was complicated by deep vein thrombosis and enterovaginal fistula. CONCLUSION: Because of the high risk of infection, women with a history of UAE and necrotic myoma adjacent to the endometrium should not undergo endometrial biopsy. We recommend evaluation of the relation of myomas to the endometrium. PMID- 18430386 TI - Genital herpes: gynaecological aspects. AB - OBJECTIVE: The purpose of this guideline is to provide recommendations to gynaecology health care providers on optimal management of genital herpes. OUTCOMES: More effective prevention of complications and transmission of genital herpes. EVIDENCE: Medline was searched for articles published in French and English related to genital herpes and gynaecology. Additional articles were identified through the references of these articles. All study types and recommendation reports were reviewed. RECOMMENDATIONS: 1. Up to 70% of all genital HSV-2 infections are transmitted during asymptomatic shedding; therefore, the use of condoms is recommended to lessen the likelihood of disease transmission. (II-A) 2. A laboratory-based diagnosis of genital herpes is essential for its effective management. (II-A) 3. Suppressive treatment is suggested for patients who have * at least 6 recurrences per year * significant complications, but fewer than 6 recurrences per year * their quality of life significantly affected * social and sexual dysfunction * to lower the risk of transmission to a sexual partner or fetus/neonate. (II-B) 4. The use of the anti viral valacyclovir, coupled with condoms and safer sex counselling, is recommended for individuals with proven genital herpes. (I-B) 5. Routine or targeted HSV screening is not indicated. PMID- 18430387 TI - Medication-induced osteoporosis. AB - Osteoporosis, a condition of low bone mass and microarchitectural deterioration, results in fractures with minimal trauma. Secondary osteoporosis is defined as bone loss resulting from either specific clinical disorders or medications. Some medications that can induce osteoporosis are discussed. Specifically, this article reviews the pathogenesis of glucocorticoid-induced bone loss and demonstrates the means to successfully manage the condition with a combination of calcium and vitamin D supplementation and, depending on the severity of the bone loss, bisphosphonates or parathyroid hormone. In addition, the pathophysiology of bone loss from aromatase inhibitors in women, gonadotropin-releasing hormone agonists in men, anticonvulsant medications, and proton pump inhibitors is outlined. Finally, this review offers suggestions on evaluation and management of bone health in individuals treated with these medications for prolonged times. PMID- 18430388 TI - Bone biopsy in patients with osteoporosis. AB - Although rarely used to diagnose and manage patients with osteoporosis, bone biopsies are performed to establish bone quality, including degree of mineralization and microarchitecture; to assess bone turnover and bone loss mechanisms; and to analyze treatment effects on bone structure and bone turnover. Bone biopsies are also the only method to diagnose mineralization defect or frank osteomalacia. Due to the availability of antiresorptive agents and anabolic drugs, determining bone turnover and bone-loss mechanisms is critical to appropriate treatment regimen selection. Bone biopsies establish the safety and efficacy of new therapeutic modalities. Further, new techniques such as molecular morphometry (in situ hybridization and immunohistochemistry) and analysis of bone content and crystal perfection have been applied to undecalcified bone and elucidated pathogenetic mechanisms or abnormalities in bone microstructure. PMID- 18430389 TI - The role of hormone therapy and calcium plus vitamin D for reduction of bone loss and risk for fractures: lessons learned from the Women's Health Initiative. AB - Osteoporosis, a major public health problem, is characterized by increased risk for fracture. To reduce the morbidity and excess loss of life associated with this common disease, we need to understand the efficacy of treatment strategies for fracture reduction. The Women's Health Initiative Clinical Trials have extended our understanding of the effect of hormone therapy and calcium plus vitamin D supplements on risk for hip and total fractures. Although estrogen, with or without progestin, significantly decreases fracture risk at all skeletal sites-almost irrespective of underlying risk for osteoporosis-its risks outweigh its benefits, negating its general use for fracture reduction. For calcium replete women, calcium plus vitamin D supplementation has a non-significant effect, hence the case for universal supplementation loses merit. But, that argument gains credibility for women over age 60-as a 21% reduction in hip fractures attests-showing that calcium plus vitamin D has a positive effect on bone health in older postmenopausal women. PMID- 18430390 TI - Anorexia, bulimia, and the athletic triad: evaluation and management. AB - Caloric restriction caused by undernutrition or over-exercise is increasingly common and has significant health consequences such as hypothalamic amenorrhea, infertility, attainment of low peak bone mass, and bone loss leading to fracture. In these patients, the pathophysiology of amenorrhea and bone loss is multifactorial, involving hormones that integrate the nutritional state with the hypothalamic-pituitary-ovarian axis, including leptin and possibly ghrelin. The pathophysiology of bone loss includes nutritional deficiencies, possibly estrogen deficiency, and direct and indirect effects of leptin on bone. Identifying patients at risk for low bone mineral density and fracture is important, as is screening with dual energy radiograph absorptiometry. Treatment has focused on oral contraceptive use, yet improved bone mineral density is marked by nutritional recovery and anovulation reversal. Therefore, resolving the nutrition deficiency should be the cornerstone of treatment. Cognitive-behavioral therapy aims for weight recovery, which can lead to reversal of amenorrhea and improvement in other associated metabolic abnormalities. During treatment, estradiol levels can be followed to assess hypothalamic-pituitary-ovarian recovery because estradiol secretion may increase well before ovulation occurs. In patients failing the above interventions, hormone replacement should be considered, but bone mineral density should be followed because patients may continue to lose bone despite treatment with oral contraceptives if nutrition is not improved. PMID- 18430391 TI - Etidronate: what is its place in treatment of primary osteoporosis and other demineralizing diseases today? AB - Bisphosphonate drugs are the major treatment options for primary and secondary osteoporosis and other demineralizing bone diseases. This class of drugs was presaged over a decade ago when etidronate disodium, the "mother compound" for modern-day bisphosphonates, was first used in the treatment of osteoporosis. The cyclic use of etidronate in therapy, which is known mainly to specialists in the field, is not approved in the United States. The drug does, however, have a worldwide reputation as a relatively inexpensive, efficacious, and highly tolerable treatment for osteoporosis. Many studies still describe its use for primary osteoporosis and some have described use in immobilization bone loss, periprosthetic bone loss, and even glucocorticoid-induced osteoporosis. This review highlights some of these uses. PMID- 18430392 TI - Bone loss in patients with breast or prostate cancer. AB - Cancers of the breast and prostate are very common in the general population, with breast cancer accounting worldwide for 23% of cancer cases in women and prostate cancer accounting for 12% of cases in men. During the past decade, the survival rates of patients with estrogen-dependent breast cancer and testosterone dependent prostate cancer have improved. This improvement has been possible thanks to the introduction of hormone treatments that suppress the synthesis or antagonize the actions of gonadal steroids. However, estrogen and testosterone deficiencies are associated with excessive bone resorption that translates into damage of the bone microarchitecture, loss of bone mineral density, and predisposition to osteoporosis and fractures. Herein, we review the mechanisms of bone loss in breast and prostate cancer survivors, their clinical implications, and different available therapeutic modalities that may help to correct the damage of bone and prevent the development of fractures. PMID- 18430393 TI - Over-suppression of bone turnover: does it exist? AB - Bone-turnover suppression is central to the therapeutic benefit of many interventions used to prevent osteoporotic fractures. There are theoretical concerns that long-term suppression may have adverse effects on bone strength, although at present no direct evidence exists that this happens. Nevertheless, further research is required to establish the optimal duration of treatment with antiresorptive agents. PMID- 18430394 TI - Vitamin D therapy. AB - The fat-soluble vitamin D prohormones, ergocalciferol (vitamin D2) and cholecalciferol (vitamin D3), are essential for the efficient intestinal absorption of calcium and phosphate and the subsequent mineralization of bone. Inadequate vitamin D leads to chronic secondary hyperparathyroidism and osteoporosis. The increasing prevalence of osteoporosis has paralleled a pandemic of vitamin D insufficiency. Based on observational and prospective trials with clinical end points, the standards for vitamin D sufficiency have been recently revised. All patients with osteopenia or osteoporosis should be monitored with a reliable assay to maintain serum 25-hydroxyvitamin D levels more than 32 ng/dL. Patients who are taking bisphosphonates and those with coexisting primary hyperparathyroidism are not exempt from taking supplemental vitamin D. PMID- 18430395 TI - Safety of parathyroid hormone for the treatment of osteoporosis. AB - Teriparatide (recombinant human 1-34 parathyroid hormone) has been registered for the treatment of postmenopausal osteoporosis and osteoporosis in men for more than 5 years, whereas 1-84 parathyroid hormone has just recently been registered in Europe for osteoporosis management. Therefore, more data are available regarding the long-term safety of teriparatide. The issues to be considered are the effects of the registered dose of teriparatide (20 microg/day) on the incidence of hypercalcemia, hypercalciuria, and hyperuricemia, and the US Food and Drug Administration's "black-box" warning regarding osteogenic sarcoma in the rat model. This review discusses these issues and provides the author's extensive clinical experience and advice on the use of teriparatide in clinical practice. PMID- 18430396 TI - Intravenous zoledronic acid for the treatment of osteoporosis. AB - Osteoporosis is a preventable disease characterized by loss of bone mineral density, progression to diminished skeletal integrity, extensive bone fragility, and an increased risk of fracture. Morbidity and mortality from osteoporosis are significant, and the public health costs are substantial. Although safe and effective therapy is available, the disease remains underdiagnosed and undertreated. Oral bisphosphonates such as alendronate, risedronate, and ibandronate are the current first-line therapy; however, adherence to treatment is suboptimal, largely because the treatment regimen is difficult to follow. The bisphosphonate zoledronic acid (zoledronate), 5 mg, is administered as an annual 15-minute intravenous infusion for the treatment of postmenopausal osteoporosis, assuring treatment compliance and persistence over the 12-month dosing interval. The drug is safe and generally well tolerated, and provides sustained anti fracture efficacy at all relevant skeletal sites. Zoledronic acid has the potential to improve clinical outcomes by reducing the risk of fracture in patients with osteoporosis. PMID- 18430397 TI - Combination anabolic and antiresorptive therapy for osteoporosis: opening the anabolic window. AB - Antiresorptive agents for osteoporosis are a cornerstone of therapy, but anabolic drugs have recently increased our options. By stimulating bone formation, anabolic agents reduce fracture incidence by improving bone qualities in addition to increasing bone mass. The only anabolic agent currently approved for osteoporosis by the US Food and Drug Administration, teriparatide (recombinant human parathyroid hormone ), has emerged as a major approach to selected patients with osteoporosis. Recombinant human parathyroid hormone (1-84) is also available in Europe. Teriparatide increases bone density and bone turnover, improves microarchitecture, and changes bone size. The incidence of vertebral and nonvertebral fractures is reduced. A current concept in the mechanism of teriparatide action is related to its effect to stimulate processes associated with bone formation before it stimulates processes associated with bone resorption. This sequence of events has led to the concept of the anabolic window, the period of time when teriparatide is maximally anabolic. Newer approaches to the use of teriparatide alone and in combination with antiresorptive agents have led to ways in which the anabolic window can be expanded. PMID- 18430398 TI - Osteonecrosis of the jaws and bisphosphonates. AB - Osteonecrosis of the jaws associated with bisphosphonate therapy was first identified in 2003 as a condition typified by exposed bone that does not heal after 8 weeks. Other signs and symptoms, such as pain and infection, may or may not be present. There is a strong need for consensus on a case definition for this condition. This condition has occurred primarily among cancer patients treated with the aminobisphosphonates zoledronic acid and pamidronate. The etiology of this condition remains unknown; however, oral disease and trauma appear to be important risk factors. This condition appears to be rare in metabolic bone disease and Paget's disease, with an estimated prevalence of approximately 1 per 100,000 person-years. Thus, the benefits of bisphosphonate therapy appear to outweigh the risks. Recommendations have been established for the oral health management of patients with a history of bisphosphonate therapy. PMID- 18430399 TI - Treatment of premenopausal women with low bone mineral density. AB - Interpretation of bone mineral density (BMD) results in premenopausal women is particularly challenging, because the relationship between BMD and fracture risk is not the same as for postmenopausal women. Z scores rather than T scores should be used to define "low BMD" in premenopausal women. The finding of low BMD in a premenopausal woman should prompt an evaluation for secondary causes of bone loss. If a secondary cause is found, management should focus on treatment of this condition. In some cases in which the secondary cause cannot be addressed, such as glucocorticoid therapy or cancer chemotherapy, treatment with a bone-active agent to prevent bone loss should be considered. In women with no fractures and no known secondary cause, low BMD may not signify compromised bone strength. BMD is likely to remain stable in these women, and pharmacologic therapy is rarely justified. Assessment of markers of bone turnover and follow-up bone density measurements can help to identify those with an ongoing process of bone loss that may indicate a higher risk for fracture, and possible need for pharmacologic intervention. PMID- 18430400 TI - The use of proton NMR as an alternative for the amino acid analysis as identity test for peptides. AB - Proton NMR was evaluated as an alternative to amino acid analysis as an identity test for peptides. Proton NMR can readily distinguish and identify all peptides currently described in the European Pharmacopoeia (Ph. Eur.). A comparison with amino acid analysis as an identity test is presented. PMID- 18430401 TI - Glycan mapping of glycotherapeutics. AB - Glycosylation of proteins may, or may not, affect biological activity, either directly or indirectly. In addition, variability in glycosylation arises within the manufacturing procedure. Therefore, glycosylation would be an important parameter when assessing product quality. Regulatory authorities require biotechnological and biological products to be characterised, including determination of their biological activity, physicochemical and immunochemical properties, their purity and their impurity profiles. Here we outline how we can decide if and which types of glycan analysis and methodologies to use for glycoprotein therapeutics to answer the scientific questions as well as meeting regulatory requirements that already exist or are going to be developed/implemented. PMID- 18430402 TI - Trace determination of ethyl methanesulfonate in Viracept 250 mg tablets by gas chromatography-mass spectrometry. AB - Ethyl methanesulfonate (EMS) is a potential human mutagenic and carcinogenic compound which has been found by Roche laboratories in nelfinavir mesylate, the active pharmaceutical ingredient of Viracept. In order to verify the quality of the medicinal product, a gas chromatographic method using mass spectrometry detection was developed for the trace analysis of EMS in Viracept 250 mg tablets from Roche laboratories. Combined with suitable sample preparation including a liquid/liquid extraction this method allows the EMS quantification with a reporting limit of 5 ppm. The extract is injected on a gas chromatographic system with a CP624-CB capillary column. Selected Ion Monitoring mode was used for the EMS quantification. Some validation elements of the method are reported. The validation study was performed over a range from 5 ppm to 100 ppm. PMID- 18430403 TI - Assessment of the need to coat particle collection cups of the NGI to mitigate droplet bounce when evaluating nebuliser-produced droplets. AB - The European Pharmaceutical Aerosol Group (EPAG) has been undertaking a series of investigations in support of developing robust methods for the droplet size analysis of nebuliser-produced aerosols for the proposed European Pharmacopoeia general chapter 2.9.44 covering preparations for nebulisation. The present two centre study, which was designed to confirm whether or not coating the collection cups of the Next Generation pharmaceutical Impactor (NGI, the recommended impactor for the proposed monograph) is required, forms part of these investigations. The study evaluated NGI-measured aerodynamic droplet-size distributions of aerosols of a generic salbutamol solution for nebulisation generated by two different jet nebulisers. The results demonstrated that there was no indication of biased stage deposition when silicone oil was used to coat the impaction surfaces, as compared to uncoated surfaces. The conclusion was that there is no need to coat the NGI cups when nebulised products are investigated. PMID- 18430404 TI - Cooling the NGI - an approach to size a nebulised aerosol more accurately. AB - The European Pharmaceutical Aerosol Group (EPAG) has undertaken investigations with the aim of developing robust methods for the droplet size analysis of nebuliser-produced aerosols in support of the proposed European Pharmacopeia general chapter 2.9.44 covering preparations for nebulisation. A multi-centre study was designed to investigate the effects of cooling the Next Generation pharmaceutical Impactor (NGI) before sample collection, as a means of reducing bias and variability caused by heat transfer-related evaporation. Droplets containing salbutamol were sized from 3 different nebulisers chosen to offer fundamentally different modes of aerosol generation: AeroNeb Go, a vibrating mesh nebuliser; PARI LC Plus, a breath-enhanced jet nebuliser; and MistyMax, a constant-output jet nebuliser. Each laboratory undertook determinations at ambient temperature, using an NGI pre-cooled in a refrigerator (5 degrees C for at least 90 min). The corresponding measurements were made using an ambient NGI as a benchmark. Salbutamol solution 5 mg/2 ml (Teva, Runcorn, UK) was used throughout the study. Analysis of individual and pooled results from 5 of the participants showed a similar trend insofar as the cooled NGI yielded a coarser nebulised aerosol than that obtained by the ambient NGI. Mass Median Aerodynamic Diameter (MMAD) was on average reduced by 9.5-21.9 % and the Fine Droplet Fraction < 5 microm (FDF) increased on average by 5.5-17.4 % for all the nebuliser designs when comparing ambient to cooled NGI. Despite the more laborious procedure of cooling the NGI, variability in data was generally similar to that obtained with the ambient NGI. We conclude that it is beneficial to cool the NGI when sizing nebulised aerosol. Furthermore, occasional findings during this study revealed a build-up of solute deposits within the interior of the NGI, and a more rigorous impactor cleaning/drying procedure is therefore recommended. PMID- 18430405 TI - Position paper: recommendation on the adoption of breathing patterns for infants and small children in general chapter 2.9.44. Preparations for nebulisation. AB - The treatment of infants and small children with nebuliser-based therapy is commonplace. However, the breathing pattern proposed for general chapter 2.9.44 covering preparations for nebulisation, in development for the European Pharmacopoeia, is that of an adult at rest. This position paper is a concise summary of recommendations for 3 breathing patterns that would be applicable to a neonate, a 12-month-old infant, and a child approximately 4 years old. The information presented herein is intended to inform the development of the proposed general chapter with the intention that information be provided to assist those involved with the evaluation of preparations for paediatric use in the choice of more appropriate breathing patterns for the assessment of active substance delivery rate and total active substance delivered. PMID- 18430406 TI - Nuclear receptors and human disease: resistance to thyroid hormone and lipodystrophic insulin resistance. PMID- 18430407 TI - [Intestinal metabolism and control of the appetite: from genes to diseases]. PMID- 18430408 TI - Thyroid hormone analogues: useful biological probes and potential therapeutic agents. PMID- 18430409 TI - Inhibitory effect of acetyl-11-keto-beta-boswellic acid on androgen receptor by interference of Sp1 binding activity in prostate cancer cells. AB - Androgen receptor (AR)-mediated signaling is crucial for the development and progression of prostate cancer (PCa). Naturally occurring phytochemicals that target the AR signaling offer significant protection against this disease. Acetyl 11-keto-beta-boswellic acid (AKBA), a compound isolated from the gum-resin of Boswellia carterii, caused G1-phase cell cycle arrest with an induction of p21(WAF1/CIP1), and a reduction of cyclin D1 as well in prostate cancer cells. AKBA-mediated cellular proliferation inhibition was associated with a decrease of AR expression at mRNA and protein levels. Furthermore, the functional biomarkers used in evaluation of AR transactivity showed suppressions of prostate-specific antigen promoter-dependent and androgen responsive element-dependent luciferase activities. Additionally, down-regulation of an AR short promoter mainly containing a Sp1 binding site suggested the essential role of Sp1 for the reduction of AR expression in cells exposed to AKBA. Interruption effect of AKBA on Sp1 binding activity but not Sp1 protein levels was further confirmed by EMSA and transient transfection with a luciferase reporter driven by three copies of the Sp1 binding site of the AR promoter. Therefore, anti-AR properties ascribed to AKBA suggested that AKBA-containing drugs could be used for the development of novel therapeutic chemicals. PMID- 18430410 TI - Deregulated expression of pro-survival and pro-apoptotic p53-dependent genes upon Elongator deficiency in colon cancer cells. AB - Elongator, a multi-subunit complex assembled by the IkappaB kinase-associated protein (IKAP)/hELP1 scaffold protein is involved in transcriptional elongation in the nucleus as well as in tRNA modifications in the cytoplasm. However, the biological processes regulated by Elongator in human cells only start to be elucidated. Here we demonstrate that IKAP/hELP1 depleted colon cancer-derived cells show enhanced basal expression of some but not all pro-apoptotic p53 dependent genes such as BAX. Moreover, Elongator deficiency causes increased basal and daunomycin-induced expression of the pro-survival serum- and glucocorticoid-induced protein kinase (SGK) gene through a p53-dependent pathway. Thus, our data collectively demonstrate that Elongator deficiency triggers the activation of p53-dependent genes harbouring opposite functions with respect to apoptosis. PMID- 18430411 TI - Representational flexibility and specificity following spatial descriptions of real-world environments. AB - Current theories are mixed with regard to the nature of mental representations following spatial description reading. Whereas some findings argue that individuals' representations are invariant following text-based, map-based, or first-person experience, other studies have suggested that representations can also exhibit considerable flexibility. In the current project we investigated the influences of spatial description perspectives and depictions on the nature of mental representations. In Experiment 1, participants exhibited more flexibility following survey, compared to route, spatial descriptions. With extended study time, though, flexibility following route descriptions increased. In Experiment 2, complementary maps further enhanced flexibility for route-based descriptions. Interestingly, increased exposure to these maps actually reduced flexibility following survey descriptions. These results demonstrate that the nature of our spatial mental representations depends upon a variety of factors; delineating these factors is critical for resolving debates concerning the malleable and invariant characteristics of spatial memory. PMID- 18430412 TI - Tissue-engineered bone formation with cryopreserved human bone marrow mesenchymal stem cells. AB - Bone marrow mesenchymal stem cells (MSCs) have become the main cell source for bone tissue engineering. It has been reported that cryopreserved human MSCs can maintain their potential for proliferation and osteogenic differentiation in vitro. There are, however, no reports on osteogenesis with cryopreserved human MSCs in vivo. The aim of this study was to determine whether cryopreservation had an effect on the proliferation capability and osteogenic differentiation of human MSCs on scaffolds in vitro and in vivo. MSCs were isolated from human bone marrow, cultured in vitro until passage 2, and then frozen and stored at -196 degrees C in liquid nitrogen with 10% Me(2)SO as cryoprotectant for 24h. The cryopreserved MSCs were then thawed rapidly, seeded onto partially demineralized bone matrix (pDBM) scaffolds and cultured in osteogenic media containing 10mM sodium beta-glycerophosphate, 50muM l-ascorbic acid, and 10nM dexamethasone. Non cryopreserved MSCs seeded onto the pDBM scaffolds were used as control groups. Scanning electronic microscopy (SEM) observation, DNA content assays, and measurements of alkaline phosphatase (ALP) activity and osteocalcin (OCN) content were applied, and the results showed that the proliferation potential and osteogenic differentiation of MSCs on pDBM in vitro were not affected by cryopreservation. After 2 weeks of subculture, the MSCs/pDBM composites were subcutaneously implanted into the athymic mice. The constructs were harvested at 4 and 8 weeks postimplantation, and histological examination showed tissue engineered bone formation in the pDBM pores in both groups. Based on these results, it can be concluded that cryopreservation allows human MSCs to be available for potential therapeutic use to tissue-engineer bone. PMID- 18430413 TI - Investigations on the heat transport capability of a cryogenic oscillating heat pipe and its application in achieving ultra-fast cooling rates for cell vitrification cryopreservation. AB - Theoretically, direct vitrification of cell suspensions with relatively low concentrations ( approximately 1 M) of permeating cryoprotective agents (CPA) is suitable for cryopreservation of almost all cell types and can be accomplished by ultra-fast cooling rates that are on the order of 10(6-7) K/min. However, the methods and devices currently available for cell cryopreservation cannot achieve such high cooling rates. In this study, we constructed a novel cryogenic oscillating heat pipe (COHP) using liquid nitrogen as its working fluid and investigated its heat transport capability to assess its application for achieving ultra-fast cooling rates for cell cryopreservation. The experimental results showed that the apparent heat transfer coefficient of the COHP can reach 2 x 10(5) W/m(2).K, which is two orders of the magnitude higher than traditional heat pipes. Theoretical analyzes showed that the average local heat transfer coefficient in the thin film evaporation region of the COHP can reach 1.2 x 10(6) W/m(2).K, which is approximately 10(3) times higher than that achievable with standard pool-boiling approaches. Based on these results, a novel device design applying the COHP and microfabrication techniques is proposed and its efficiency for cell vitrification is demonstrated through numerical simulation. The estimated average cooling rates achieved through this approach is 10(6-7)K/min, which is much faster than the currently available methods and sufficient for achieving vitrification with relatively low concentrations of CPA. PMID- 18430414 TI - The Drosophila wing hearts originate from pericardial cells and are essential for wing maturation. AB - In addition to the heart proper, insects possess wing hearts in the thorax to ensure regular hemolymph flow through the narrow wings. In Drosophila, the wing hearts consist of two bilateral muscular pumps of unknown origin. Here, we present the first developmental study on these organs and report that the wing hearts originate from eight embryonic progenitor cells arising in two pairs in parasegments 4 and 5. These progenitors represent a so far undescribed subset of the Even-skipped positive pericardial cells (EPC) and are characterized by the early loss of tinman expression in contrast to the continuously Tinman positive classical EPCs. Ectopic expression of Tinman in the wing heart progenitors omits organ formation, indicating a crucial role for Tinman during progenitor specification. The subsequent postembryonic development is a highly dynamic process, which includes proliferation and two relocation events. Adults lacking wing hearts display a severe wing phenotype and are unable to fly. The phenotype is caused by omitted clearance of the epidermal cells from the wings during maturation, which inhibits the formation of a flexible wing blade. This indicates that wing hearts are required for proper wing morphogenesis and functionality. PMID- 18430415 TI - Control of cell cycle timing during C. elegans embryogenesis. AB - As a fundamental process of development, cell proliferation must be coordinated with other processes such as fate differentiation. Through statistical analysis of individual cell cycle lengths of the first 8 out of 10 rounds of embryonic cell division in Caenorhabditis elegans, we identified synchronous and invariantly ordered divisions that are tightly associated with fate differentiation. Our results suggest a three-tier model for fate control of cell cycle pace: the primary control of cell cycle pace is established by lineage and the founder cell fate, then fine-tuned by tissue and organ differentiation within each lineage, then further modified by individualization of cells as they acquire unique morphological and physiological roles in the variant body plan. We then set out to identify the pace-setting mechanisms in different fates. Our results suggest that ubiquitin-mediated degradation of CDC-25.1 is a rate-determining step for the E (gut) and P(3) (muscle and germline) lineages but not others, even though CDC-25.1 and its apparent decay have been detected in all lineages. Our results demonstrate the power of C. elegans embryogenesis as a model to dissect the interaction between differentiation and proliferation, and an effective approach combining genetic and statistical analysis at single-cell resolution. PMID- 18430416 TI - Involvement of HMG-12 and CAR-1 in the cdc-48.1 expression of Caenorhabditis elegans. AB - Caenorhabditis elegans possesses two p97/VCP/Cdc48p homologues, named CDC-48.1 (C06A1.1) and CDC-48.2 (C41C4.8), and their expression patterns and levels are differently regulated. To clarify the regulatory mechanisms of differential expression of two p97 proteins of C. elegans, we performed detailed deletion analysis of their promoter regions. We found that the promoter of cdc-48.1 contains two regions necessary for embryonic and for post-embryonic expression, while the promoter of cdc-48.2 contains the single region necessary for embryonic expression. In particular, two elements (Element A and Element B) and three conserved boxes (Box a, Box b and Box c) were essential for cdc-48.1 expression in embryos and at post-embryonic stages, respectively. By using South-Western blotting and MALDI-TOF MS analysis, we identified HMG-12 and CAR-1 as proteins that bind to Element A and Element B, respectively, from the embryonic nuclear extract. Importantly, we found the decreased expression of p97 in embryos prepared from hmg-12(RNAi) or car-1(RNAi) worms. These results indicate that both HMG-12 and CAR-1 play important roles in embryonic expression of cdc-48.1. PMID- 18430417 TI - Two deltaC splice-variants have distinct signaling abilities during somitogenesis and midline patterning. AB - Notch signaling is required for many developmental processes, yet differences in the signaling abilities of various Notch ligands are poorly understood. Here, we have isolated a splice variant of the zebrafish Notch ligand deltaC in which the inclusion of the last intron leads to a truncation of the C-terminal 39 amino acids (deltaC(tv2)). We show that, unlike deltaC(tv1), deltaC(tv2) cannot function effectively in somitogenesis but has an enhanced ability to signal during midline development. Additionally, over-expression of deltaC(tv2) preferentially affects anterior midline development, while another Notch ligand, deltaD, shows a posterior bias. Using chimeric Deltas we show that the intracellular domain is responsible for the strength of signal in midline development, while the extracellular domain influences the anterior-posterior bias of the effect. Together our data show that different deltas can signal in biologically distinct ways in both midline formation and somitogenesis. Moreover, it illustrates the importance of cell-type-dependent modifiers of Notch signaling in providing ligand specificity. PMID- 18430418 TI - Fluvastatin decreases cardiac fibrosis possibly through regulation of TGF beta(1)/Smad 7 expression in the spontaneously hypertensive rats. AB - Statins ameliorate myocardial fibrosis after myocardial infarction. We designed this study to determine whether fluvastatin reduced hypertension-induced myocardial hypertrophy and fibrosis and whether these fluvastatin effects involved transforming growth factor beta1 (TGF-beta1) and Smad 7, factors known to play a role in the myocardial hypertrophy and fibrosis. We randomized 14 week old spontaneously hypertensive rats (SHRs) to receiving vehicle or 5-20 mg/kg/day fluvastatin for 8 weeks. Wistar Kyoto (WKY) rats receiving vehicle or 10 mg/kg/day fluvastatin were also studied. SHRs had an increased blood pressure, left ventricular hypertrophy and fibrosis compared with WKY rats. SHRs also had an elevated TGF-beta1 expression and a decreased Smad 7 expression. These changes in SHRs were dose-dependently attenuated by fluvastatin. For example, the hydroxyproline content was 3.2+/-0.1, 4.0+/-0.1 and 3.5+/-0.1 microg/mg heart and the Smad 7 protein expression was 5.1+/-0.6, 1.0+/-0.1 and 4.1+/-0.7 arbitrary units for WKY rats, SHRs and SHRs receiving 20 mg/kg/day fluvastatin, respectively. The hydroxyproline content in the SHRs treated with or without fluvastatin was positively correlated with the left ventricular mass index, systolic blood pressure and the amount of TGF-beta1 proteins and negatively correlated with the Smad 7 expression level. The left ventricular mass index was positively correlated with the systolic blood pressure. Fluvastatin did not alter the blood pressure, left ventricular mass index and collagen content of WKY rats. These results suggest that fluvastatin reduces hypertension-induced myocardial hypertrophy and fibrosis. These effects may involve an increased expression of Smad 7 and a decreased expression of TFG-beta1. Our results call for clinical studies to evaluate these fluvastatin effects in hypertensive patients. PMID- 18430419 TI - Ginsenoside Rg1 protects neurons from hypoxic-ischemic injury possibly by inhibiting Ca2+ influx through NMDA receptors and L-type voltage-dependent Ca2+ channels. AB - The purpose of this study is to assess the neuroprotective effect of Rg1, a ginsenoside. We measured cell viability and lactate dehydrogenase (LDH) release from primary culture of rat hippocampal neurons and electrical activities in hippocampal slices of rats, before and after the neurons were deprived of oxygen and glucose. In addition, cerebral damage was evaluated with magnetic resonance imaging after middle cerebral artery was occluded transiently. Nissl staining was used for histological observation and immunohistochemistry analysis for activated caspase-3 expression of the brain. Furthermore, calcium influx was measured with laser confocal microscopy in neurons perfused with KCl (50 mM) or N-methyl-d aspartate (NMDA, 1 mM), or deprived of oxygen and glucose. The influences of ginsenoside Rg1 on these parameters were determined simultaneously. We found that treatment of Rg1: 1) increased the neuronal viability; 2) promoted the recovery of electrical activity in hippocampal slices; 3) reduced the release of LDH, cerebral damage area, neuronal loss and expression of caspase-3; and 4) inhibited calcium influx induced by NMDA, KCl or oxygen/glucose deprivation. However, the protective effect of Rg1 was blocked by mifepristone, an antagonist of glucocorticoid receptors. Taken together, these results suggest that ginsenoside Rg1 can reduce neuronal death, including apoptotic cell death, induced by hypoxic ischemic insults. This neuroprotective effect is probably mediated by the activation of glucocorticoid receptors, and by the inhibition of calcium influx through NMDA receptors and L-type voltage-dependent Ca2+ channels and the resultant reduction of intracellular free Ca2+. PMID- 18430420 TI - Subacute hypoxia suppresses Kv3.4 channel expression and whole-cell K+ currents through endogenous 15-hydroxyeicosatetraenoic acid in pulmonary arterial smooth muscle cells. AB - We have previously reported that subacute hypoxia activates lung 15-lipoxygenase (15-LOX), which catalyzes arachidonic acid to produce 15-HETE, leading to constriction of neonatal rabbit pulmonary arteries. Subacute hypoxia suppresses Kv3.4 channel expression and results in an inhibition of whole-cell K(+) currents (I(K)). Although the Kv channel inhibition is likely to be mediated through 15 HETE, direct evidence is still lacking. To reveal the role of the 15-LOX/15-HETE pathway in the hypoxia-induced down-regulation of Kv3.4 channel expression and inhibition of I(K), we performed studies using 15-LOX blockers, whole-cell patch clamp, semi-quantitative PCR, ELISA and Western blot analysis. We found that Kv3.4 channel expression at the mRNA and protein levels was greatly up-regulated in pulmonary arterial smooth muscle cells after blockade of 15-LOX by CDC or NDGA. The 15-LOX blockade also partially restored I(K). In comparison, 15-HETE had a stronger effect than 12-HETE on the expression of Kv3.4 channels. 5-HETE had no noticeable effect on Kv3.4 channel expression. These data indicate that the 15-LOX pathway via its metabolite, 15-HETE, seems to play a role in the down regulation of Kv3.4 expression and I(K) inhibition after subacute hypoxia. PMID- 18430421 TI - Schistosoma mansoni: SmE78, a nuclear receptor orthologue of Drosophila ecdysone induced protein 78. AB - Drosophila ecdysone-induced protein 78 (E78) belongs to the nuclear receptor (NR) superfamily, it plays a role directly related to ecdysone signaling. We isolated a cDNA of Drosophila E78 orthologue from the Platyhelminth Schistosoma mansoni (SmE78). It is the first E78 orthologue known outside of the molting animals--the Ecdysozoa. The SmE78 cDNA is 3471 base pairs long and contains an entire open reading frame (ORF) encoding a 1087 amino acid protein. Phylogenetic analysis of the ligand-binding domain (LBD) demonstrates that the LBD of SmE78 is phylogenetically related to the Drosophila E78. Gene structure of SmE78 was determined showing it to consist of six exons spanning more than 32 kbp. Quantitative real-time reverse-transcriptase polymerase chain reaction (RT-PCR) demonstrated that SmE78 was expressed throughout schistosome development but with the highest levels of expression in miracidia and egg stage. The result is consistent with the previous studies that Ecdysterone was effective in stimulating host location activities in miracidia. The data suggest that SmE78 may be involved in transduction of an ecdysone signal in S. mansoni. PMID- 18430422 TI - The roles of the alpha1-adrenergic receptor subtypes in rat embryonic implantation. AB - OBJECTIVE: To focus on the possible roles of alpha(1)-adrenergic receptors (alpha(1)-ARs) in rat embryonic implantation. DESIGN: Laboratory study. SETTING: Animal and pharmacology laboratory at Department of Pharmacodynamics and Biopharmacy, University of Szeged, Hungary. ANIMAL(S): Pregnant and nonpregnant Sprague-Dawley rats. INTERVENTION(S): Uterus tissues were collected during the peri-implantation period. MAIN OUTCOME MEASURE(S): We used a reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting to demonstrate the expressions of mRNAs and the protein expressions of the alpha(1) AR subtypes in the early-pregnant uterus. Electric field stimulation was applied to test the pharmacologic reactivity of the alpha(1A)-AR, and the physiologic role of this receptor was tested in a knock-down transformed animal model using an antisense oligonucleotide that elicits sequence-selective inhibition of the alpha(1A)-AR gene expression. RESULT(S): The presence of all alpha(1)-AR subtypes (alpha(1A), alpha(1B), and alpha(1D)) was proved, with a predominance of alpha(1A)-AR. The maximal expression of the alpha(1A)-AR was attained on the day of implantation. The selective alpha(1A) antagonist 5-methylurapidil inhibited the contraction in a dose-dependent manner. The number of implantation sites was decreased ( approximately 75%) in the alpha(1A)-AR knock-down transformed rats. CONCLUSION(S): We assume that the alpha(1A)-AR predominance plays a crucial role in embryonic implantation in the rat. PMID- 18430423 TI - Estrogenic exposure affects metamorphosis and alters sex ratios in the northern leopard frog (Rana pipiens): identifying critically vulnerable periods of development. AB - During the transformation from larval tadpole to juvenile frog, there are critical periods of metamorphic development and sex differentiation that may be particularly sensitive to endocrine disruption. The aim of the present study was to identify sensitive developmental periods for estrogenic endocrine disruption in the northern leopard frog (Rana pipiens) using short, targeted exposures to the synthetic estrogen, ethinylestradiol (EE2). Post-hatch tadpoles (Gosner stage 27) were exposed over five distinct periods of metamorphosis: early (stage 27 30), mid (stage 30-36), early and mid (stage 27-36), late (stage 36-42), and the entire metamorphic period (chronic; stage 27-42). For each period, animals were sampled immediately following the EE2 exposure and at metamorphic climax (stage 42). The effects of EE2 on metamorphic development and sex differentiation were assessed through measures of length, weight, developmental stage, days to metamorphosis, sex ratios and incidence of gonadal intersex. Our results show that tadpoles exposed to EE2 during mid-metamorphosis were developmentally delayed immediately following exposure and took 2 weeks longer to reach metamorphic climax. In the unexposed groups, there was low proportion (0.15) of intersex tadpoles at stage 30 and gonads appeared to be morphologically distinct (male and female) in all individuals by stage 36. Tadpoles exposed early in development displayed a strong female-biased sex ratio compared to the controls. Moreover, these effects were also seen at metamorphic climax, approximately 2-3 months after the exposure period, demonstrating that transient early life-stage exposure to estrogen can induce effects on the reproductive organs that persist into the beginning of adult life-stages. PMID- 18430424 TI - Distinct cytochrome P450 aromatase isoforms in the common carp (Cyprinus carpio): sexual dimorphism and onset of ontogenic expression. AB - Cytochrome P450 aromatase (CYP19) is a key enzyme in the steroidogenic pathway that catalyses the conversion of testosterone to estrogen, and therefore is thought to influence gonadal sex differentiation. In an effort to understand the role of this enzyme in ovarian differentiation, we isolated cDNA encoding the two distinct isoforms, ovarian and brain (termed cyp19a and cyp19b, respectively) of adult common carp, Cyprinus carpio. The cloned cDNA for cyp19a had an open reading frame (ORF) of 518 amino acid residues, in contrast to cyp19b with an ORF of 511 amino acids. Sequence and phylogenetic analysis showed that these CYP19 isoforms were orthologous with previously described cyp19a and cyp19b from other teleosts. Quantitative real-time PCR indicated that both isoforms are expressed in adult ovary and brain, with predominant expression of cyp19a in the ovary and cyp19b in the brain. The major aromatase expressing tissue was found to be the brain, with greatest cyp19b expression in the anterior quarter (telencephalon) in both sexes. The gonad showed sexually dimorphic expression of both genes and dimorphic expression of cyp19a was observed in the cerebellum and the liver. Ontogenic expression showed that only the ovarian aromatase transcript is inherited maternally, with lower expression observed through early larval development under warmer rearing conditions. The differential and overlapping expression suggests these two aromatase genes have different roles in reproductive physiology. PMID- 18430425 TI - Non-invasive measurement of adrenocortical and gonadal activity in male and female guinea pigs (Cavia aperea f. porcellus). AB - Taking blood samples is a common method in biomedical and biological research using guinea pigs. However, most blood sampling techniques are complicated and highly invasive and may therefore not be appropriate for certain research topics concerning stress and reproduction. Thus, a non-invasive method to measure steroid hormones is critically needed. The aim of this study was the biological validation of corresponding enzyme immunoassays for the measurement of fecal cortisol, progesterone, estrogen, and testosterone metabolites in guinea pigs. We examined the effect of subcutaneous injections of ACTH or saline on fecal cortisol metabolites to investigate the suitability of fecal samples to monitor adrenocortical activity. Furthermore, we investigated whether fecal sex steroid metabolites accurately reflected endocrine changes observed in plasma samples during female estrous cycles and male puberty, respectively. In addition, we compared fecal testosterone metabolites of intact males, castrated males, and females to investigate the reliability of fecal samples in discriminating gonadal status of males. Concentrations of fecal cortisol metabolites were significantly increased following ACTH challenge, indicating that adrenocortical activity can be monitored via fecal samples. Secondly, in females, plasma and fecal gonadal steroids were significantly correlated in most subjects. The assay for testosterone metabolites, on the other hand, could not clearly discriminate between test groups. From these findings we conclude that fecal samples can be used for the non-invasive assessment of adrenocortical and female reproductive status in guinea pigs. Testosterone metabolism seems to be more complex and further investigations are needed to establish a more suitable assay. PMID- 18430426 TI - Incomplete nonsense-mediated mRNA decay in Giardia lamblia. AB - Messenger RNAs containing premature translation stop codons are degraded by a nonsense-mediated mRNA decay (NMD) system. The NMD pathway is present in yeast, plants and mammals and is thought to protect cells from production of nonfunctional proteins by rapidly degrading mutant mRNAs. There is little understanding of the biology of the origins of eukaryotes, particularly of the NMD pathway. Searches using the BLAST program revealed that the protozoan Giardialamblia has only some of the components of the NMD pathway. We developed a luciferase reporter system with a nonsense mutation to monitor NMD in Giardia. The nonsense mutation triggered a decrease in luciferase mRNA levels and stability, suggesting that the NMD phenomenon could be present in Giardia. We also found a significant reduction of the mRNA levels of another system containing Giardia its own cyst wall protein 3 gene with a nonsense mutation. However, the reduction levels observed in these two systems are lower than that in late-branching eukaryotes, suggesting that the NMD system in Giardia may be less functional. Interestingly, the effect of G418 in promoting read-through of the nonsense mutation and inhibiting NMD in Giardia is similar to that in late branching eukaryotes. We also characterised the giardial homologue of a conserved NMD factor, UPF1. Immunofluorescence assays revealed that giardial UPF1, like yeast UPF1, is expressed in the cytoplasm, but not in the nucleus. In addition, overexpression of UPF1 resulted in a reduction of the levels of nonsense containing transcripts and enhanced translation termination at a nonsense codon. These results suggest that Giardia may have an incomplete NMD pathway and giardial UPF1 may be functionally conserved, involved in NMD and in preventing nonsense suppression. PMID- 18430427 TI - Fluctuation of lipoprotein metabolism linked with bile acid-activated liver nuclear receptors in Alagille syndrome. AB - Alagille syndrome (AGS) is a rare hereditary disorder exhibiting fluctuating cholestasis and dyslipidemia. Farnesoid X receptor (FXR) and liver X receptor (LXR) are hepatic nuclear receptors that regulate bile acid and lipoprotein metabolism. To investigate whether cholestasis is related to dyslipidemia and hepatic nuclear receptor expression in AGS patients, we determined the blood levels of total bile acid (TBA) and lipoprotein parameters, and examined hepatic nuclear receptor expression in three AGS children and their three incomplete AGS parents repeatedly over several years. In the AGS children, TBA level showed significant positive correlations with low-density lipoprotein-cholesterol, apolipoprotein E (apoE)-rich high-density lipoprotein-cholesterol (HDL-C), apoA I, apoE, and cholesteryl ester transfer protein (CETP) concentrations, but negative correlation with apoE-poor HDL-C concentration. Western blot analysis of liver biopsy specimens revealed that FXR and LXR expression increased in parallel with TBA level. CETP- and ATP-binding cassette transporter A1 expression also increased with TBA level, while scavenger receptor class B type-I expression showed the opposite response. However, apoA-I expression was similar to the control level at any TBA level. In the incomplete AGS parents, TBA and lipoprotein parameters showed little fluctuation. In summary, cholestasis is closely related to dyslipidemia and hepatic nuclear receptor expression in AGS patients. PMID- 18430428 TI - Co-administration of berberine and plant stanols synergistically reduces plasma cholesterol in rats. AB - The objective of the present study was to determine the beneficial effects and the safety of oral administration of the combination of berberine (BBR) and plant stanols (PS) on plasma lipid profiles in male Sprague-Dawley rats. Four groups of animals were fed a cornstarch-casein-sucrose-based high-cholesterol (2%, w:w) and high-fat (27.5%) diet. Three treatment groups were supplemented with either BBR (100mgkg(-1)bodyweightd(-1)), PS (1% in diet, w:w), or the combination of both (BBRPS). After 6 wk, animals were sacrificed and followed immediately with the collection of blood and organ samples. Lipid analysis revealed that PS lowered plasma total cholesterol (T-C) by 18% (p=0.067) and non-HDL-cholesterol (non-HDL C) by 29% (p=0.013) as compared with the control, while BBR had no effect on both T-C and non-HDL-C. The combination treatment of BBRPS reduced plasma T-C by 41% (p=0.0002) and non-HDL-C by 59% (p<0.0001) compared to the control group. BBR reduced plasma TG levels by 31% at a marginal significance relative to the control (p=0.054), whereas PS had no effect. BBRPS showed an additive effect of BBR and PS on plasma TAG. PS and BBRPS both decreased liver cholesterol (p=0.0027 and 0.0002, respectively). BBR and PS, either alone or in combination, did not show any toxic effects as assessed by plasma concentration of hepatic biochemical parameters. These results demonstrate that BBR and PS, when combined, synergistically lower plasma cholesterol levels and significantly reduce liver cholesterol, without the observation of any toxic effects. PMID- 18430429 TI - On the problems of describing joint axis alignment. AB - Each three-dimensional joint possesses at least one potentially oblique axis of rotation. Several systems are used to express joint axis alignment. One system, designated the plane projection (PP) method, describes angles based on orthogonal projections onto two, of the three, anatomical planes. Alternatively, a joint axis may be described in two different ways using two sequential Cardan angle rotations. These expression systems all lay claim to similar descriptive labels, such as deviation and elevation. Difficulties arise as researchers use these various methods to compare their own data to the results of others. A joint axis alignment, described as 27 degrees deviation and 41 degrees elevation in PP, differs by as much as 6 degrees when expressed as Cardan angles. Differences among expression systems increase as the joint axis alignment becomes more oblique -- eventually differing by as much as 75 degrees . This paper explores implications for this lack of congruence among the joint axis expression systems. Effective steps in dealing with these issues begin with recognizing the existence and extent of the problem. The paper provides a common set of algorithms to illustrate and alleviate the possible problems associated with the exchange of joint axis alignment data. PMID- 18430430 TI - Rinse and evaporation coating of poly(methyl methacrylate) microchip for separation of sodium dodecyl sulfate-protein complex. AB - We developed a novel channel wall coating on a poly(methyl methacrylate) (PMMA) microchip using methylcellulose (MC) as a coating reagent to suppress electroosmotic flow (EOF) following the strong analytes adsorption via hydrophobic interaction with channel walls of PMMA. Our coating was obtained by first rinsing channel walls with MC-containing aqueous solution followed by evaporation. The coating made the hydrophilic channel wall lowering EOF by two orders of magnitude (1.2 x 10(-5)cm(2)V(-1)s(-1)) as well as reducing the hydrophobic adsorption. On the coated channel walls, we successfully separated sodium dodecyl sulfate-protein complexes with high reproducibility and efficiency using dextran as a lower viscosity protein separation medium. PMID- 18430431 TI - Facile fabrication of multi-colors high fluorescent/superparamagnetic nanoparticles. AB - We developed a novel method to prepare multi-colors high fluorescent/superparamagnetic nanoparticles (FMNPs) employing hydrophobic multi color quantum dots (QDs) and hydrophobic Fe(3)O(4) (MNPs) via ultrasonic emulsification method. This structural procedure was simple, one-off, and timesaving. Different-sizes FMNPs with encoding single/multi-color QDs and MNPs were achieved. Analysis with transmission electron microscopy (TEM) and particle size analyzer demonstrated that the as-prepared samples were spherical, uniform in size distribution; Ultraviolet-visible (UV-vis) absorption spectroscopy and photoluminescence (PL) measurement showed the FMNPs had good optical properties, lacking of fluorescence resonance energy transfer (FRET) inside FMNPs; vibrating sample magnetometer (VSM) indicated that FMNPs were superparamagnetic. These results indicate that the as-prepared FMNPs have potential of serving as a hybrid of QDs and MNPs in bioanalysis communities. PMID- 18430432 TI - Interaction of mineral surfaces with simple organic molecules by diffuse reflectance IR spectroscopy (DRIFT). AB - The adsorption of simple organic molecules to mineral surfaces was investigated by Diffuse Reflectance Infrared Fourier Transform Spectroscopy (DRIFTS) and by solution chemical analysis. Salicylic acid, myristic acid or octacosane were deposited from water or hexane onto gamma-alumina and kaolinite, and examined without evacuation or blending with KBr. The DRIFTS results replicated those from ATR-IR, where available, supporting the validity of DRIFTS as a versatile and convenient alternative to ATR. Results were obtained for coverage as low as 0.02 molecules/nm(2). Monolayer coverage on gamma-alumina was determined from spectra as 0.7 molecules/nm(2) for salicylic acid, confirmed by solution analysis, and 2 molecules/nm(2) for myristic acid. Spectral features of salicylic acid deposited on gamma-alumina from hexane were not distinguishable from those deposited from water. In almost all cases, spectral features associated with the carboxyl moiety were substantially altered, indicating a significant role in the adsorption mechanism. Adsorption of salicylic acid from hexane onto kaolinite was also as carboxylate, but myristic acid showed both carboxylate and carbonyl. The results from using hexane as solvent compared to water suggest that surface-adsorbed hydroxyl and molecular water, present on mineral oxides under ambient conditions, may be a key determinant of the adsorbate architecture. PMID- 18430433 TI - Developmental trends in everyday conditional reasoning: the retrieval and inhibition interplay. AB - Two experiments examined developmental patterns in children's conditional reasoning with everyday causal conditionals. In Experiment 1, a group of pre-, early, young, and late adolescents generated counterexamples for a set of conditionals to validate developmental claims about the counterexample retrieval capacity. In Experiment 2, participants in the same age range were presented with a conditional reasoning task with similar conditionals. Experiment 1 established that counterexample retrieval increased from preadolescence to late adolescence. Experiment 2 showed that acceptance rates of the invalid affirmation of the consequent inference gradually decreased in the same age range. Acceptance rates of the valid modus ponens inference showed a U-shaped pattern. After an initial drop from preadolescence to early adolescence, modus ponens acceptance ratings increased again after the onset of adolescence. Findings support the claim that the development of everyday conditional reasoning can be characterized as an interplay between the development of a counterexample retrieval and inhibition process. PMID- 18430434 TI - Development of a cell death-based method for the screening of nuclear factor kappaB inhibitors. AB - Nuclear factor kappa B (NF-kappaB) plays a significant role in immunity and inflammation and represents a first choice as pharmacological target for anti inflammatory therapy. However, research in this field has been hampered by the fact that no convenient assay suitable for large-scale screening procedures is available. The present study provides a cell death-based assay method for screening of nuclear factor-kappaB inhibitors. In this study, we observed that four distinct pharmacologic inhibitors of NF-kappaB, pyrrolidine dithiocarbamate (PDTC), N-tosyl-L-lysyl chloromethyl ketone (TPCK), genistein and BAY11-7082, resulted in the cell death of murine macrophages, J774A.1. DNA-binding experiments showed that lethal doses were consistent with those required for NF kappaB inhibition. DNA fragmentation analysis showed that cell death is apoptotic in nature. Further studies suggested that NF-kappaB inhibitors induced apoptosis is independent of the involvement of other markers of cell death such as caspases and p38 MAP (Mitogen activated protein) kinase. From this study, we conclude that NF-kappaB activation may represent an important survival mechanism in macrophages. This study also provides a new cell-based screening method, as any compound that will inhibit NF-kappaB activity will result in the death of macrophages. PMID- 18430435 TI - Immune response affects ant trophallactic behaviour. AB - Sociality is associated with many benefits that have favoured its evolution in social insects. However, sociability also presents disadvantages like crowding of large numbers of individuals, which may favour the spread of infections within colonies. Adaptations allowing social insects to prevent and/or control pathogen infections range from behavioural responses to physiological ones including their immune systems. In a state of infection, social interactions with nestmates should be altered in a way which might prevent its spreading. We simulated a microbial infection in workers of the ant Camponotus fellah by the administration of peptidoglycan (PGN) and then quantified their immune response and social interactions. PGN injections as well as control injections of Ringer solution elicited similar production of antibacterial compounds, during 1-4 days after. However, injections of PGN reduced the ability of encapsulation of a nylon implant compared to Ringer controls. The immune challenged workers did not decrease the level of interactions with their nestmates. On the contrary, they devoted more time to trophallaxis. These results are discussed in relation to ant life history traits. PMID- 18430436 TI - Studies on adaptations of Metarhizium anisopliae to life in the soil. AB - Metarhizium anisopliae is an important fungal model for elucidating the basis of entomopathogenicity. A field trial conducted in 2000 with a strain expressing the gfp gene as a marker unexpectedly identified the rhizosphere (the root-soil interface) as the site where insects and pathogen most likely interact. However, the implications of rhizosphere events in maintaining large populations of M. anisopliae remain unclear. The extent to which plant ecology is impacted by the rhizosphere competence of M. anisopliae is also uncertain, but it could be considerable with implicit co-evolutionary implications. Rhizosphere competence may need to be considered as a feature for selecting fungal strains for biocontrol and this raises the possibility of managing the rhizosphere microflora to achieve insect control. Rhizosphere competence also raises some concerns, particularly if it resulted in introduced or hypervirulent transgenic strains persisting in the environment. It is hoped that field trials and other studies currently underway to extend our knowledge of host-pathogen-plant interactions will help identify containment strategies as well as lead to new and important ways of exploiting insect pathogenic fungi for crop protection. PMID- 18430438 TI - ErbB receptors, their ligands, and the consequences of their activation and inhibition in the myocardium. AB - The epidermal growth factor (EGF) receptor (or ErbB1) and the related ErbB4 are transmembrane receptor protein tyrosine kinases which bind extracellular ligands of the EGF family. ErbB2 and ErbB3 are "co-receptors" structurally related to ErbB1/ErbB4, but ErbB2 is an "orphan" receptor and ErbB3 lacks tyrosine kinase activity. However, both are important in transmembrane signalling. All ErbB receptors/ligands are intimately involved in the regulation of cell growth, differentiation and survival, and their dysregulation contributes to some human malignancies. After extracellular ligand binding, receptor dimerisation and transautophosphorylation of intracellular C-terminal tyrosine residues, they bind signalling proteins which recognise specific tyrosine-phosphorylated motifs. This leads to activation of multiple signalling pathways, notably the extracellular signal-regulated kinase 1/2 (ERK1/2) cascade and the phosphoinositide 3-kinase (PI3K)/protein kinase B [PKB/(Akt)] pathway. In heart, targeted deletion of ErbB2, ErbB3, ErbB4 and some ErbB receptor extracellular ligands leads to embryonic lethality resulting from cardiovascular defects. ErbB receptor ligands improve cardiac myocyte viability and are hypertrophic, partly because of activation of ERK1/2 and/or PI3K/PKB(Akt). Furthermore, ErbB transactivation by Gq protein-coupled receptor (GqPCR) signalling may mediate the hypertrophic effects of GqPCR agonists. The utility of anthracyclines in cancer chemotherapy can be limited by their cardiotoxic side effects and these may be counteracted by ErbB receptor ligands. ErbB2 is the target of anti-cancer monoclonal antibody trastuzumab (Herceptin), and its myocardial downregulation may account for the occasional cardiotoxicity of this therapy. Here, we review the basic biochemistry of ErbB receptors/ligands, and emphasise their particular roles in the myocardium. PMID- 18430437 TI - Bone marrow-derived pluripotent very small embryonic-like stem cells (VSELs) are mobilized after acute myocardial infarction. AB - The adult bone marrow (BM) harbors Sca-1+/Lin-/CD45- pluripotent very small embryonic-like stem cells (VSELs), which can differentiate in vitro into several lineages, including cardiac and vascular lineages. Since mobilization of stem/progenitors from the BM is a prerequisite for their participation in organ repair, we investigated whether VSELs are mobilized into the peripheral blood (PB) after acute myocardial infarction (MI). Wild-type mice (C57BL/6 strain, 6- or 15-wk-old) underwent a 30-min coronary occlusion followed by reperfusion (groups III-V, VIII-X, n=6-12/group) or a 1-hour open-chest state (sham controls, groups II and VII, n=8-12/group); mice were sacrificed 24 h, 48 h, or 7 days later and PB samples were harvested. Controls (groups I and VI, n=6/group) were sacrificed without any intervention. By flow cytometry, VSELs were barely detectable in PB under baseline conditions but their levels increased significantly at 48 h after MI, both in younger (6-wk-old) and older (15-wk-old) mice (3.33+/-0.37 and 7.10+/-0.89 cells/microl of blood, respectively). At 48 h after MI, qRT-PCR analysis revealed significantly increased levels of mRNA of markers of pluripotency (Oct-4, Nanog, Rex-1, Rif1, and Dppa1) in PB cells of 6 wk-old (but not 15-wk-old) infarcted mice compared with either controls or sham controls. Confocal microscopy and ImageStream analysis confirmed that mobilized VSELs expressed Oct-4 protein, while Sca-1+/Lin-/CD45+ hematopoietic stem cells did not. This is the first demonstration that Oct-4+ pluripotent stem cells (VSELs) are mobilized from the BM into the PB after acute MI. This phenomenon may have pathophysiological and therapeutic implications for repair of infarcted myocardium. PMID- 18430439 TI - Data compression and genomes: a two-dimensional life domain map. AB - We define the complexity of DNA sequences as the information content per nucleotide, calculated by means of some Lempel-Ziv data compression algorithm. It is possible to use the statistics of the complexity values of the functional regions of different complete genomes to distinguish among genomes of different domains of life (Archaea, Bacteria and Eukarya). We shall focus on the distribution function of the complexity of non-coding regions. We show that the three domains may be plotted in separate regions within the two-dimensional space where the axes are the skewness coefficient and the curtosis coefficient of the aforementioned distribution. Preliminary results on 15 genomes are introduced. PMID- 18430440 TI - Expiration rate drives human airway design. AB - Analyses of human airway architecture based on calculations of airflow resistance or energy dissipation suggest that the branching pattern is not optimized for minimizing energy loss by flow dissipation during respiration. Airway flow dissipates only a few percent of the total body work during normal breathing, so branching patterns deviate from minimum energy loss to also optimize other physiological needs. Studies of airway performance often record some measure of expiration, such as FEV1 (Forced Expiratory Volume in 1s), because airway constriction during expiration limits the rate of rapid respiration. We posit that lung structure is optimized for the rate of expiration as well as minimum energy loss. By increasing the daughter-to-parent airway diameter ratio (h) from 0.794 (corresponding to the energy minimum for symmetrically branching airways) to 0.85 (the observed value in humans) luminal pressures at airway generations 4 15 were substantially increased during exercise (a 4.5 and 15 cmH2O increase during moderate and heavy exercise, respectively). Values of h somewhat larger than 0.794 help airways remain open during expiration by increasing both viscous pressure drop and convective acceleration pressure drop. Asymmetric bifurcations also exhibit higher proximal airway pressures than symmetric ones, but the improvement was not large. PMID- 18430441 TI - [Positron emission tomography: which indications, which benefits?]. AB - Positron emission tomography (PET) is currently used in the presurgical workup for drug-resistant partial epilepsies in addition to MRI. Interictal metabolism is studied in clinical practice using (18)fluoro-desoxy-glucose ((18)FDG). In medial temporal lobe epilepsy (MTLE) associated with hippocampal sclerosis, hypometabolism ipsilateral to the epileptogenic focus is found in 70-90% of cases. However, hypometabolism is larger than the structural lesion observed on MRI and includes the epileptogenic zone and ictal discharge spread areas. Hypometabolism is related to surgical outcome and cognitive disturbances in MTLE. Although the usefulness of PET appears less well-established in extratemporal lobe epilepsy and in children, its sensitivity may be improved by coregistration and superimposition of PET on MRI at any age. Focal hypometabolism can be easily detected by visual analysis, allowing detection of minor gyral abnormalities that may correspond to focal cortical dysplasias. Moreover, in cases of negative MRI, focal hypometabolism findings may help invasive monitoring planning and deep electrode placement for SEEG, and finally improve surgical outcome. PMID- 18430442 TI - Effects of isolation-rearing on the development of social behaviors in male Mongolian gerbils (Meriones unguiculatus). AB - Social isolation has been shown to modify behavioral patterns and neuroendocrine function in many species. In this study, we examined the influence of post weaning social isolation on the behavioral development and fecal testosterone and corticosterone levels of male Mongolian gerbils. In Experiment 1, isolation reared (IR) males spent more time engaged in social-sniffing. IR males also showed more aggressive and anxiety-related behaviors than group-reared males (GR; three animals per cage). In Experiment 2, a screen-divided-reared condition (SDR) was examined in addition to GR (two males per cage) and IR males. In the SDR group, two subjects were separated by a wire mesh screen allowing for sensory communication and limited physical contact but not direct social interactions. SDR males showed similar patterns in sniffing and anxiety-related behaviors to IR males; however, an increase in aggression was not observed. No differences in fecal testosterone and corticosterone levels were found among males in different rearing conditions. These results suggest that post-weaning social isolation affects behavioral development without affecting testosterone and corticosterone levels. The behavioral changes observed may therefore be regulated by different mechanisms; increases in social-sniffing and anxiety-related behaviors may be caused by the deprivation of direct social interactions, whereas the enhancement of aggressive behaviors may be caused by the lack of sensory stimulation and limited physical contact with other individuals. PMID- 18430443 TI - Epidemiological evaluation of Chagas disease in a rural area of southern Bolivia. AB - We evaluated the prevalence of Chagas disease using a rapid screening test (Chagas Stat-Pak), confirmed by ELISA, in Carapari, a village of 9000 inhabitants in southern Bolivian Chaco. The prevalence of Trypanosoma cruzi was estimated in a sample of 995 people. The prevalence adjusted on age was 51.2% and was proportionally related to age. We also observed a very significant cline from the south to the north of the locality, where the prevalence ranged from 40 to 80%. In children younger than 11 years, the prevalence was 21.5%, which confirmed the importance of residual vector transmission despite several years of vector control. Among women of procreation age, the prevalence was 63.9%, resulting in a high risk of congenital transmission. The control of the disease requires an increase in vector control and improvement of dwellings before considering children's treatment with trypanocide. PMID- 18430444 TI - Poverty and musculoskeletal impairment in Rwanda. AB - The recently adopted UN Convention on the Rights of Persons with Disabilities acknowledges the need to address social exclusion and poverty of persons with disabilities. However, policy makers, especially in low-income countries, often lack information about the socioeconomic situation of this vulnerable group of society. This study aimed to assess the association between poverty and musculoskeletal impairment (MSI) in Rwanda. A nationwide population-based matched case-control study was undertaken in Rwanda. Data were collected on education, literacy, employment, household expenditure and assets for 345 cases and 532 matched controls. Conditional logistic regression was performed, and the results indicated that adults with MSI in Rwanda are more likely to have no employment (odds ratio (OR)=3.3, 95% CI 2.1-5.2) while children with MSI are less likely to attend school (OR=0.4, 95% CI 0.2-0.9). Cases with MSI are disadvantaged vis-a vis housing conditions and household size, potentially indicating crowding. However, cases with MSI were not poorer than controls in terms of assets or expenditure. These data suggest that increased efforts should be undertaken in Rwanda in order to ensure that children with disabilities are included in schools and that adults with disabilities can find appropriate employment opportunities. PMID- 18430445 TI - Induction of Rad51 protein levels by p38 MAPK decreases cytotoxicity and mutagenicity in benzo[a]pyrene-exposed human lung cancer cells. AB - Rad51 is an essential component of the homologous recombination repair pathway. Abnormal expression of Rad51 has been reported in various carcinomas. Benzo[a]pyrene (B[a]P), a polycyclic hydrocarbon carcinogen found in the environment, induces cancer in multiple organs. B[a]P has been shown to activate the p38 MAPK signaling pathway in mammalian cells. The prime purpose of this study was to determine how B[a]P activates the p38 MAPK signaling pathway, and how this then regulates Rad51 expression in human cancer cells. Exposure of human lung cancer cells with B[a]P increased Rad51 protein levels in a time- and dose dependent fashion. B[a]P also induced Rad51 mRNA and protein synthesis. Blockage of p38 MAPK activation by SB202190 or small interfering RNA (si-p38) decreased B[a]P-elicited Rad51 protein levels by increasing Rad51 protein instability, but did not affect Rad51 mRNA transcription. Furthermore, enhancement of p38 MAPK signaling by constitutively active MKK6 (MKK6E) increased Rad51 protein levels and protein stability. Moreover, B[a]P-induced cytotoxicity and mutagenicity were significantly increased in cells depleted of endogenous Rad51. Taken together, these results indicate that Rad51 protein provides a critical role in inhibiting the cytotoxicity and mutagenicity of B[a]P in B[a]P-treated human lung cancer cells. Furthermore, the work points to an unexpected role of p38 MAPK signaling in the control of Rad51 protein stability in response to B[a]P exposure. PMID- 18430446 TI - The effects of subchronic acrylamide exposure on gene expression, neurochemistry, hormones, and histopathology in the hypothalamus-pituitary-thyroid axis of male Fischer 344 rats. AB - Acrylamide (AA) is an important industrial chemical that is neurotoxic in rodents and humans and carcinogenic in rodents. The observation of cancer in endocrine responsive tissues in Fischer 344 rats has prompted hypotheses of hormonal dysregulation, as opposed to DNA damage, as the mechanism for tumor induction by AA. The current investigation examines possible evidence for disruption of the hypothalamic-pituitary-thyroid axis from 14 days of repeated exposure of male Fischer 344 rats to doses of AA that range from one that is carcinogenic after lifetime exposure (2.5 mg/kg/d), an intermediate dose (10 mg/kg/d), and a high dose (50 mg/kg/d) that is neurotoxic for this exposure time. The endpoints selected include: serum levels of thyroid and pituitary hormones; target tissue expression of genes involved in hormone synthesis, release, and receptors; neurotransmitters in the CNS that affect hormone homeostasis; and histopathological evaluation of target tissues. These studies showed virtually no evidence for systematic alteration of the hypothalamic-pituitary-thyroid axis and do not support hormone dysregulation as a plausible mechanism for AA-induced thyroid cancer in the Fischer 344 rat. Specifically, there were no significant changes in: 1) mRNA levels in hypothalamus or pituitary for TRH, TSH, thyroid hormone receptor alpha and beta, as well 10 other hormones or releasing factors; 2) mRNA levels in thyroid for thyroglobulin, thyroid peroxidase, sodium iodide symporter, or type I deiodinases; 3) serum TSH or T3 levels (T4 was decreased at high dose only); 4) dopaminergic tone in the hypothalamus and pituitary or importantly 5) increased cell proliferation (Mki67 mRNA and Ki-67 protein levels were not increased) in thyroid or pituitary. These negative findings are consistent with a genotoxic mechanism of AA carcinogenicity based on metabolism to glycidamide and DNA adduct formation. Clarification of this mechanistic dichotomy may be useful in human cancer risk assessments for AA. PMID- 18430447 TI - PON1Q192R genetic polymorphism modifies organophosphorous pesticide effects on semen quality and DNA integrity in agricultural workers from southern Mexico. AB - Pesticide exposure, including organophosphorous (OP) insecticides, has been associated with poor semen quality, and paraoxonase (PON1), an enzyme involved in OP deactivation, may have a role on their susceptibility, due to PON1 polymorphisms. Our objective was to evaluate the role of PON1Q192R polymorphism on the susceptibility to OP toxicity on semen quality and DNA integrity in agricultural workers. A cross-sectional study was conducted in farmers with Mayan ascendancy from southeastern Mexico chronically exposed to pesticides; mostly OP. Fifty four agricultural workers (18-55 years old) were included, who provided semen and blood samples. Semen quality was evaluated according to WHO, sperm DNA damage by in situ-nick translation (NT-positive cells), PON1Q192R polymorphism by real-time PCR and serum PON1 activity by using phenylacetate and paraoxon. Two OP exposure indexes were created: at the month of sampling and during 3 months before sampling, representing the exposure to spermatids-spermatozoa and to cells at one spermatogenic cycle, respectively. PON1 192R and 192Q allele frequencies were 0.54 and 0.46, respectively. Significant associations were found between OP exposure at the month of sampling and NT-positive cells and sperm viability in homozygote 192RR subjects, and dose-effect relationships were observed between OP exposure during 3 months before sampling and sperm quality parameters and NT positive cells in homozygote 192RR farmers. This suggests that cells at all stages of spermatogenesis are target of OP, and that there exists an interaction between OP exposure and PON1Q192R polymorphism on these effects; farmers featuring the 192RR genotype were more susceptible to develop reproductive toxic effects by OP exposure. PMID- 18430448 TI - A protein phosphatase 2A (PP2A) inhibition assay using a recombinant enzyme for rapid detection of microcystins. AB - Worldwide blooms of toxic cyanobacteria (blue-green algae) commonly occur in freshwater, often in drinking water sources, necessitating routine monitoring of water quality. Microcystin-LR and related cyanobacterial toxins strongly inhibit protein phosphatase 2A (PP2A) and are therefore assayable by measuring the extent of PP2A inhibition. In this study, we evaluated the suitability of the catalytic subunit of recombinant PP2A (rPP2Ac) expressed with a baculovirus system for use in a microplate microcystin assay. Five microcystin analogs, microcystin-LR, -RR, -YR, -LF, and -LW, and nodularin strongly inhibited rPP2Ac activity with IC(50) values of 0.048, 0.072, 0.147, 0.096, 0.114, and 0.54 nM, respectively. Microcystin-LR in a water sample could be assayed from 0.005 to 5 ng/ml. The assay could detect the toxin at a far lower level than required by the World Health Organization for regulation of microcystin-LR or its equivalent (1 microg/L). Pretreatment or concentration of water samples with low toxin concentrations was not necessary. The microplate assay using rPP2Ac was more sensitive than an enzyme-linked immunosorbent assay (ELISA) method and a cytotoxicity assay. The genetically engineered rPP2Ac was more stable than a commercially available dimeric enzyme, producing accurate and reproducible results. Our results confirm that the rPP2Ac we prepared is an excellent tool for detecting and quantifying microcystins in water. PMID- 18430449 TI - High-speed digital scan converter for high-frequency ultrasound sector scanners. AB - This paper presents a high-speed digital scan converter (DSC) capable of providing more than 400 images per second, which is necessary to examine the activities of the mouse heart whose rate is 5-10 beats per second. To achieve the desired high-speed performance in cost-effective manner, the DSC developed adopts a linear interpolation algorithm in which two nearest samples to each object pixel of a monitor are selected and only angular interpolation is performed. Through computer simulation with the Field II program, its accuracy was investigated by comparing it to that of bilinear interpolation known as the best algorithm in terms of accuracy and processing speed. The simulation results show that the linear interpolation algorithm is capable of providing an acceptable image quality, which means that the difference of the root mean square error (RMSE) values of the linear and bilinear interpolation algorithms is below 1%, if the sample rate of the envelope samples is at least four times higher than the Nyquist rate for the baseband component of echo signals. The designed DSC was implemented with a single FPGA (Stratix EP1S60F1020C6, Altera Corporation, San Jose, CA) on a DSC board that is a part of a high-speed ultrasound imaging system developed. The temporal and spatial resolutions of the implemented DSC were evaluated by examining its maximum processing time with a time stamp indicating when an image is completely formed and wire phantom testing, respectively. The experimental results show that the implemented DSC is capable of providing images at the rate of 400 images per second with negligible processing error. PMID- 18430450 TI - Functional analysis of bipartite begomovirus coat protein promoter sequences. AB - We demonstrate that the AL2 gene of Cabbage leaf curl virus (CaLCuV) activates the CP promoter in mesophyll and acts to derepress the promoter in vascular tissue, similar to that observed for Tomato golden mosaic virus (TGMV). Binding studies indicate that sequences mediating repression and activation of the TGMV and CaLCuV CP promoter specifically bind different nuclear factors common to Nicotiana benthamiana, spinach and tomato. However, chromatin immunoprecipitation demonstrates that TGMV AL2 can interact with both sequences independently. Binding of nuclear protein(s) from different crop species to viral sequences conserved in both bipartite and monopartite begomoviruses, including TGMV, CaLCuV, Pepper golden mosaic virus and Tomato yellow leaf curl virus suggests that bipartite begomoviruses bind common host factors to regulate the CP promoter. This is consistent with a model in which AL2 interacts with different components of the cellular transcription machinery that bind viral sequences important for repression and activation of begomovirus CP promoters. PMID- 18430451 TI - Global shape versus local feature: an angle illusion. AB - We have shown previously that the precision of angle judgments depends strongly on the global stimulus configuration: discrimination thresholds for angles that form part of isosceles triangles are up to 3 times lower than for those that form part of scalene triangles [Kennedy, G. J., Orbach, H. S., & Loffler, G. (2006). Effects of global shape on angle discrimination. VisionResearch, 46(8-9), 1530 1539]. Here, we investigated whether or not the perceived size of an angle (accuracy) is also affected by the overall shape of which it forms a part. Observers compared the relative sizes of angles contained in isosceles triangles with those of angles in scalene triangles and points of subjective equality were determined. For a reference angle of 60 degrees , angles embedded in isosceles triangles were judged to be on average 14 degrees larger than angles embedded in scalene triangles. This result is largely independent of the reference angle, triangle orientation and triangle size. Moreover, the effect is present whether or not triangles of different shapes enclose the same area, whether or not the side of the triangle opposite the angle is present and whether the triangle is outlined or defined by dots at its vertexes. In sum, our results provide evidence for a novel illusion where an angle embedded in an isosceles triangle is judged substantially larger than the same angle embedded in a scalene triangle. This finding demonstrates that mechanisms for computing angles are sensitive to the context within which angles are presented. PMID- 18430452 TI - Cloning, expression, purification, cofactor requirements, and steady state kinetics of phosphoketolase-2 from Lactobacillus plantarum. AB - The genes xpk1 and xpk2(Delta1-21) encoding phosphoketolase-1 and (Delta1-7) truncated phosphoketolase-2 have been cloned from Lactobacillus plantarum and expressed in Escherichia coli. Both gene-products display phosphoketolase activity on fructose-6-phosphate in extracts. A N-terminal His-tag construct of xpk2(Delta1-21) was also expressed in E. coli and produced active His-tagged (Delta1-7)-truncated phosphoketolase-2 (hereafter phosphoketolase-2). Phosphoketolase-2 is activated by thiamine pyrophosphate (TPP) and the divalent metal ions Mg(2+), Mn(2+), or Ca(2+). Kinetic analysis and data from the literature indicate the activators are MgTPP, MnTPP, or CaTPP, and these species activate by an ordered equilibrium binding pathway, with Me(2+)TPP binding first and then fructose-6-phosphate. Phosphoketolase-2 accepts either fructose-6 phosphate or xylulose-5-phosphate as substrates, together with inorganic phosphate, to produce acetyl phosphate and either erythrose-4-phosphate or glyceraldehyde-3-phosphate, respectively. Steady state kinetic analysis of acetyl phosphate formation with either substrate indicates a ping pong kinetic mechanism. Product inhibition patterns with erythrose-4-phosphate indicate that an intermediate in the ping pong mechanism is formed irreversibly. Background mechanistic information indicates that this intermediate is 2-acetyl-TPP. The irreversibility of 2-acetyl-TPP formation might explain the overall irreversibility of the reaction of phosphoketolase-2. PMID- 18430453 TI - Distribution and availability of arsenic in soils from the industrialized urban area of Beijing, China. AB - Concentrations of arsenic (As) were determined in soils of 5 industrial sites in an urban area of Beijing, China. Fifty seven typical surface soils were sampled to determine total concentrations of metals, pH and dissolved organic carbon (DOC). One hundred and eight deep soils were submitted to a four-step, sequential extraction to assess the relative mobility and bioavailability of As in the soil profiles. Total concentrations of As in surface soils ranged from 5.7 to 2.3 x 10(1) mg kg(-1), dw with greater concentrations inside the perimeter of the chemical plant which had greater concentrations than did other plants. 75.4% of surface soil samples in the industrial area contained concentrations of As that were greater than was considered to be the background concentration of 7.8 mg kg( 1), dw for the region. The mean concentration (9.9 mg kg(-1), dw) in the industrial soils was greater than that soils from other type of land use. Concentrations of As were significantly and negatively correlated with soil pH and DOC in industrial soils. Although mean concentration of total As in the soils from all sites were less at greater depths, the entire range from 0 to 180 cm (especially 0-80 cm) contained concentrations of As that were greater than background. Sequential extractions of soil indicated that only some surface soils had relatively great amount of extractable fraction of As. Most soils had relatively great amount of residual As. This result suggests that most arsenic in Beijing industrial soils should be immobile and of limited bioavailability. PMID- 18430454 TI - Evaluation of site-specific and seasonal variation in colonic mucosal eosinophils. AB - Several systemic disorders and gastrointestinal diseases may be associated with increased colonic mucosal eosinophils, which may vary in number throughout the normal colon. Some investigators have proposed that colonic eosinophilia reflects allergen exposure, although this hypothesis has never been validated, and values quantifying the number of mucosal eosinophils that can be regarded as a normal finding are lacking. The aims of this study were to determine the number of intramucosal eosinophils normally present throughout the colon and evaluate the relationship between colonic eosinophilia and seasonal allergen exposure. Eosinophils in the crypt epithelium and lamina propria were evaluated in 198 mucosal biopsy specimens obtained from the ascending (n = 98) and descending (n = 100) colon of patients with normal colonoscopic examinations. The cases were stratified into 12 groups, reflecting the months during which the samples were obtained, and the mean number of mucosal eosinophils was determined for each group. Daily air pollen counts were recorded, and the mean determined for each month. Fifty-five percent of mucosal biopsy specimens from the ascending colon contained eosinophils in the crypt epithelium, compared with only 5% of biopsy specimens from the descending colon (P < .001). Lamina propria eosinophils were, on average, 3 times more numerous in the ascending compared with the descending colon (P < .001). Mucosal eosinophils were slightly more numerous in samples obtained in April and May, corresponding to periods of highest pollen counts, but this relationship was not significant (P > .05). We conclude that intramucosal eosinophils are commonly present in the proximal colon but show only mild fluctuations with ambient allergen exposure. PMID- 18430455 TI - Biliary neoplasia with extensive intraductal spread associated with liver cirrhosis: a hitherto unreported variant of biliary intraepithelial neoplasia. AB - We describe the histopathologic features of 2 cases of biliary neoplasia with extensive intraductal spread arising in liver cirrhosis. The prevalence of this type of biliary neoplasia may be 0.4% from the review of 468 cases of cirrhotic liver. Histologic analysis revealed that the micropapillary proliferation of the atypical biliary epithelium composed of columnar cells with enlarged nuclei diffusely extended superficially from the septal intrahepatic bile duct to the reactive ductules associated with liver cirrhosis. Both cases exhibited prominent fibrous or sclerotic stroma near the biliary lesion. Immunohistochemical analysis revealed a characteristic cytokeratin and mucin expression pattern (CK7++, CK19++, CK20+, MUC1+/-, MUC2-, MUC5AC+, MUC6-). The tumor cytoplasm was focally positive for laminin gamma2 together with linear staining of the basement membrane. Proliferative activity confirmed by Ki67 staining was relatively high. Both patients were disease-free for 3 years after the operation. We believe that the possibility of biliary neoplasia with extensive intraductal spread should be considered to be a variant of biliary intraepithelial neoplasia. PMID- 18430456 TI - Placental histologic criteria for umbilical blood flow restriction in unexplained stillbirth. AB - Approximately 50% of stillbirths are unexplained after fetopsy and placental examination. Fatal hypoxic injury due to restriction of umbilical blood flow ("cord accident") may be causal in a subset of these stillbirths. We reviewed placental slides of 62 cases of third-trimester stillbirth from our autopsy files over a 5-year period to define criteria and estimate the frequency of cord accident as a cause of stillbirth. By correlating clinical and autopsy information-with placental gross and histologic findings-from a series of index cases with a strong presumptive evidence of cord accident, histologic criteria for cord accident were established. "Minimal histologic criteria," suggestive of cord accident, were defined as vascular ectasia and thrombosis within the umbilical cord, chorionic plate, and/or stem villi. A definitive diagnosis of cord accident required in addition regional distribution of avascular villi or villi showing stromal karyorrhexis. Of 27 stillbirth cases with a cause of death determined to be other than cord accident, only 3 (11%) met all histologic criteria for cord accident (specificity of 89%). In contrast, of 25 stillbirth cases with an unknown cause of death, a significantly larger subset (13 cases or 52%) met the criteria for cord accident (P = .0038). Thus, we find nonacute cord compression implicated in over half of "unexplained" fetal deaths. This is the first report to establish histologic criteria in the diagnosis of cord accident, the application of which could significantly reduce the proportion of unexplained third-trimester stillbirth. PMID- 18430457 TI - A case of multiple immunoglobulin G4-related periarteritis: a tumorous lesion of the coronary artery and abdominal aortic aneurysm. AB - Immunoglobulin G4 (IgG4)-related disease can occur in various organs, most of which are glandular or ductal tissues. Here, we report a case of multiple IgG4 related vascular lesions. A 63-year-old patient was found to have an abdominal aortic aneurysm and a tumorous lesion around the right coronary artery. The surgically resected aneurysmal wall and a tumorous lesion of the right coronary artery showed similar histologic features including diffuse lymphoplasmacytic infiltration, occasional eosinophils, and obliterative phlebitis. Immunohistochemically, numerous IgG4-positive plasma cells were evident within the lesions. The serum concentrations of IgG4 in the preoperative period was 456 mg/dL (reference range, <135), which decreased to 242 mg/dL 2 weeks after surgery. We made a diagnosis of multiple IgG4-related periarteritis manifesting as an abdominal aortic aneurysm and a tumorous nodule of the coronary artery. This case report suggested that IgG4-related disease can occur in the vascular system and manifest as an aneurysm or a periarterial mass lesion. PMID- 18430458 TI - Mutational analysis of caspase 1, 4, and 5 genes in common human cancers. AB - Mounting evidence indicates that deregulation of apoptosis is involved in the mechanisms of cancer development. Mutations of genes encoding caspases, the executioners of apoptosis, have been detected in human cancers, indicating inactivation of apoptosis by the mutations of caspase is an important mechanism in cancer development. The aim of this study was to see whether genes encoding human caspases 1, 4, and 5 are mutated in human cancers. We analyzed the entire coding region and all splice sites of human caspase 1, 4, and 5 genes for the detection of somatic mutations in 337 human cancers, including 103 colorectal, 54 gastric, 60 breast, 60 hepatocellular, and 60 lung carcinomas by a single-strand conformation polymorphism assay. We detected 2 (0.6%) caspase-1, 2 (0.6%) caspase 4, and 15 (4.4%) caspase-5 mutations in the 343 cancers. The mutations were detected in 11 gastric carcinomas (2 caspase-1 and 9 caspase-5 mutations), 6 colorectal carcinomas (2 caspase-4 and 4 caspase-5 mutations), 1 breast carcinoma (1 caspase-5 mutation), and 1 lung carcinoma (1 caspase-5 mutation). The mutations consisted of 11 mutations in exons and 8 mutations in noncoding sequences. The 11 mutations in the exons consisted of 3 missense, 1 silent, and 7 frameshift mutation(s). Of note, most (6/9) of the caspase-5 mutations in the coding sequences were detected in microsatellite instability (MSI)-positive cancers. These data indicate that somatic mutations of caspase-1 and caspase-4 genes are rare in common solid cancers. In addition, the data indicate that caspase-5 gene is commonly mutated in the MSI-positive cancers, and suggest that inactivation of caspase-5 may play a role in the tumorigenesis of MSI-positive cancers. PMID- 18430459 TI - Homologous recombination and maintenance of genome integrity: cancer and aging through the prism of human RecQ helicases. AB - Homologous recombination (HR) is a genetic mechanism in somatic cells that repairs DNA double-strand breaks and restores productive DNA synthesis following disruption of replication forks. Although HR is indispensable for maintaining genome integrity, it must be tightly regulated to avoid harmful outcomes. HR associated genomic instabilities arise in three human genetic disorders, Bloom syndrome (BS), Werner syndrome (WS), and Rothmund-Thomson syndrome (RTS), which are caused by defects in three individual proteins of the RecQ family of helicases, BLM, WRN, and RECQL4, respectively. Cells derived from persons with these syndromes display varying types of genomic instability as evidenced by the presence of different kinds of chromosomal abnormalities and different sensitivities to DNA damaging agents. Persons with these syndromes exhibit a variety of developmental defects and are predisposed to a wide range of cancers. WS and RTS are further characterized by premature aging. Recent research has shown many connections between all three proteins and the regulation of excess HR. Here, we illustrate the elaborate networks of BLM, WRN, and RECQL4 in regulating HR, and the potential mechanistic linkages to cancer and aging. PMID- 18430460 TI - Atmospheric inputs of Ag and Tl to the Arctic: comparison of a high resolution snow pit (AD 1994-2004) with a firn (AD 1860-1996) and an ice core (previous 16,000 yr). AB - Applying strict clean room techniques and sector field ICP-MS, a total of 567 ice and snow samples from Devon Ice Cap, Arctic Canada, have been analysed for silver (Ag) and thallium (Tl), providing a continuous record of atmospheric deposition of both elements for the past approximately 16,000 yr. Concentrations of Ag and Tl ranged from 0.02 to 5.84 pg/g and from 0.03 to 3.40 pg/g, respectively. The natural Tl/Sc background ratio established with ice samples dating between 10,590 and 15,010 yr BP, amounted to 0.11+/-0.03 which is in good agreement with the corresponding value of 0.107 reported for the Upper Continental Crust (UCC). The Ag/Sc background ratio in ice (0.09+/-0.06), in turn, is much more variable and approximately 10-times higher than UCC values. The high temporal resolution provided by 45 samples from the 5 m snow pit representing the period AD 1994 to 2004, revealed the greatest elemental concentrations as well as accumulation rates during the winter months when air masses reach the Arctic predominantly from Eurasia. The greatest Sc concentrations, however, are also found during winter months which clearly reflect strong seasonal variations in atmospheric dust inputs. Enrichment factors calculated for both elements (Ag, Tl) using Sc show contrasting behaviours, with the maximum EF for Tl (up to 48) during summer when air masses arriving to the Arctic are predominantly from North America, probably reflecting coal burning, base metal mining and smelting. The greatest enrichments of Ag (up to 17), on the other hand, tend generally to be found in winter, although some enrichment maxima corresponding to summer months are also seen. While modern Ag EF are comparable to those corresponding to ancient (back to ca. 3 k yr) anthropogenic activities, modern Tl EF clearly exceed the impact of Medieval, Roman and Greek/Phoenician civilization. Accumulation rates during the past decade have been variable (mean 36 and 130 ng/m(2)/yr for Ag and Tl, respectively), but the trend is clearly in decline. Based on the best estimate of natural inputs, more than 80% of the Ag and 90% of the Tl supplied to the Arctic today (AD 1994-2004) are derived from anthropogenic sources. PMID- 18430461 TI - Bioavailability as an issue in risk assessment and management of food cadmium: a review. AB - The bioavailability of cadmium (Cd) from food is an important determinant of the potential risk of this toxic element. This review summarizes the effects of marginal deficiencies of the essential nutrients zinc (Zn), iron (Fe), and calcium (Ca) on the enhancement of absorption and organ accumulation and retention of dietary Cd in laboratory animals. These marginal deficiencies enhanced Cd absorption as much as ten-fold from diets containing low Cd concentrations similar to that consumed by some human populations, indicating that people who are nutritionally marginal with respect to Zn, Fe, and Ca are at higher risk of Cd disease than those who are nutritionally adequate. Results from these studies also suggest that the bioavailability of Cd is different for different food sources. This has implications for the design of food safety rules for Cd in that if the dietary source plays such a significant role in the risk of Cd, then different foods would require different Cd limits. Lastly, the importance of food-level exposures of Cd and other potentially toxic elements in the study of risk assessment are emphasized. Most foods contain low concentrations of Cd that are poorly absorbed, and it is neither relevant nor practical to use toxic doses of Cd in experimental diets to study food Cd risks. A more comprehensive understanding of the biochemistry involved in the bioavailability of Cd from foods would help resolve food safety questions and provide the support for a badly needed advance in international policies regarding Cd in crops and foods. PMID- 18430462 TI - Monitoring aspirin treatment in patients with thrombocytosis: comparison of the platelet function analyzer (PFA)-100 with optical aggregometry. AB - Aspirin provides satisfactory protection against thrombotic episodes in essential thrombocythemia (ET), but at higher platelet counts has been less effective. Our aim was to compare the platelet function analyzer (PFA)-100 with optical aggregometry in order to determine a reliable method in monitoring aspirin's influence on platelet function in patients with thrombocytosis. We studied 36 patients with thrombocytosis. Sixteen of them, receiving aspirin, composed group A, while group B consisted of 20 patients not taking aspirin. In all patients, we compared the platelet function measured by classic optical aggregation tests with closure times (CT) obtained by the PFA-100. The definition of platelet responses as normal or pathological showed that PFA-100 collagen and/or epinephrine (CEPI) CTs and epinephrine-induced aggregometry is the pair of methods with the higher agreement in monitoring of platelet dysfunction due to ASA treatment (a=94%). Satisfactory results were also obtained for group B (a=81%). The comparison between PFA-100 CEPI CTs and arachidonic acid-induced aggregometry exhibited moderate agreement both in the total number of patients and in group A (a=79% and 94%, respectively). PFA-100 collagen and/or ADP (CADP) CTs and ADP-induced aggregometry were not concordant. The PFA-100 system appears to be a reliable and rapid method in the assessment of aspirin's antiplatelet effect in patients with thrombocytosis. Regarding aggregometry, the selection of the inducer, its concentration and cut-off points is crucial in defining the response to antiaggregating agents. It still remains to determine whether there is any relevance between the measurements obtained by these methods and clinical outcome in thrombocythemic patients. PMID- 18430465 TI - Microsurgery for recurrent lumbar disk herniation at the same level and side: do patients fare worse? Experience with 95 consecutive cases. AB - BACKGROUND: In reviewing our experience with reoperation of RLDH, our aim was mainly to determine whether patients fared worse than after primary surgery. We found no uniform answers to this question in the literature. METHODS: The data of 95 patients (29 women and 66 men) who underwent reoperation for RLDH at the same level and side were analyzed retrospectively. Forty-two patients underwent the first operation in our clinic (recurrence rate, 2.6% of 1586 cases). Gadolinium enhanced MRI was performed in all patients. Main clinical data of patients, pain free interval, operation time, surgical complications, duration of hospital stay, and clinical improvement rate were recorded. RESULTS: The mean pain-free interval was 55 months (range, 3-120 months). Levels of recurrent herniation were L4 through L5 and L5 through S1 (65% and 35% of cases, respectively). Revision surgery lasted longer on average than the previous diskectomy (P < .01) and was complicated by dural tear in 4 cases (4.2% vs 0.9% during primary diskectomy, P < .05). There were no significant differences between revision and previous surgery in terms of hospital stay. However, rates of excellent/good outcomes were significantly less for RLDH (89% vs 95%, P < .05); and the percentage of poor results was higher (2% vs 0.5%, P < .05). Age, sex, smoking, profession, trauma, level and degree of herniation, and pain-free interval were not correlated with clinical outcome. CONCLUSION: Conventional microsurgery for RLDH showed lightly but significantly worse results than those of primary microdiskectomy. Patients contemplating reoperation should be informed of this fact and of the risk of dural tear and prolonged operation time. PMID- 18430467 TI - Re: The challenge for neurosurgery in the 21st century (Ausman J. Surg Neurol 2008;69:102). PMID- 18430468 TI - Validation of FACT/GOG-AD subscale for ovarian cancer-related abdominal discomfort: a Gynecologic Oncology Group study. AB - OBJECTIVE: To evaluate the Functional Assessment of Cancer Therapy/Gynecologic Oncology Group-Abdominal Discomfort (FACT/GOG-AD) subscale for assessing abdominal discomfort in patients undergoing treatment for ovarian cancer. METHODS: A four-item questionnaire was developed for use in Gynecologic Oncology Group (GOG) protocol 172. It was administered to patients with advanced ovarian cancer treated on this protocol with intravenous (IV) cisplatin/paclitaxel or a combination of intravenous and intraperitoneal (IP) cisplatin/paclitaxel (IV/IP) prior to randomization, before cycle 4, 3-6 weeks after cycle 6, and 12 months after cycle 6. The subscale was evaluated in patients receiving IV/IP therapy for internal consistency, concurrent validity, sensitivity to treatment differences, and responsiveness to abdominal discomfort grading. RESULTS: Internal consistency coefficients were 0.83 and 0.87 at baseline and pre-cycle 4 assessments, respectively; the average inter-item correlation was 0.61 at the pre-cycle 4 assessment. Item correlation with other scales ranged from 0.00-0.44. This subscale was able to distinguish those graded to have physician-rated abdominal pain compared to those without clinician-reported discomfort. CONCLUSION: The four-item AD subscale reliably and validly assesses ovarian cancer-specific abdominal discomfort, and captures abdominal symptom responses to IV and IV/IP cisplatin/paclitaxel treatments. PMID- 18430469 TI - Epigenetic mechanisms mediating the long-term effects of maternal care on development. AB - The long-term consequences of early environmental experiences for development have been explored extensively in animal models to better understand the mechanisms mediating risk of psychopathology in individuals exposed to childhood adversity. One common feature of these models is disruption of the mother-infant relationship which is associated with impairments in stress responsivity and maternal behavior in adult offspring. These behavioral and physiological characteristics are associated with stable changes in gene expression which emerge in infancy and are sustained into adulthood. Recent evidence suggests that these long-term effects may be mediated by epigenetic modification to the promoter regions of steroid receptor genes. In particular, DNA methylation may be critical to maternal effects on gene expression and thus generate phenotypic differentiation of offspring and, through effects on maternal behavior of offspring, mediate the transmission of these effects across generations. In this review we explore evidence for the influence of mother-infant interactions on the epigenome and consider evidence for and the implications of such epigenetic effects for human mental health. PMID- 18430471 TI - Can fine particulate matter explain the paradoxical ozone associations? AB - Our previous paper entitled "Paradoxical ozone associations could be due to methyl nitrite from combustion of methyl ethers or esters in engine fuels" (Env. Int.. 2007;33;1090) reviewed 11 studies of the impact of ozone on human health that, paradoxically, found a negative coefficient for ozone-morbidity associations. We argued that the most likely explanation for this effect would be methyl nitrite (MN) as an unsuspected exhaust component of engines with methyl ether in the fuel. The basis of the argument was the fact that MN is rapidly destroyed by sunlight, so that MN would be negatively correlated with ozone. All (but one) of the reviewed studies concluded that criterion pollutants could not explain the negative slope. The argument was strengthened by the observation that such paradoxical ozone associations have not been found in regions without significant methyl ether in gasoline. Left unaddressed in the previous paper was the possibility that fine particulate matter (FPM) might explain the POA. If this were true, then it would be necessary that the FPM be negatively correlated with ozone in those regions that found a POA. The current paper reviews data on FPM ozone correlations in those regions where a POA was identified. The results show that FPM was, in most cases, positively correlated with ozone and so could not explain the POA. PMID- 18430472 TI - EBV transformation overrides gene expression patterns of B cell differentiation stages. AB - EBV-associated Hodgkin lymphoma (HL) and some post-transplant lymphoproliferative disease (PTLD) cases originate from pro-apoptotic germinal center (GC) B cells that have acquired destructive somatic Ig V gene mutations and were presumably rescued from apoptosis by EBV. To find out whether B cell receptor-crippled GC B cells acquire features of HL and/or PTLD cells upon EBV-infection and to reveal the impact of EBV on expression of B cell differentiation markers, we compared lymphoblastoid cell lines (LCLs) from GC B cells (including BCR-crippled GC-LCLs) to monoclonal LCLs from naive B cells (N-LCLs). In addition, we analyzed the controversially discussed effect of EBV-infection on the GC B-cell-specific process of somatic hypermutation in vitro. Irrespective of their cellular origin, LCLs expressed CD20, CD30, CD38, AID, Pu.1, and with one exception Syk, but lacked expression of the GC B cell marker BCL-6. Interestingly, the T cell transcription factor GATA-3 that is aberrantly expressed in HL was induced in most GC-LCLs and the memory B cell marker CD27 was activated in N-LCLs. Remarkably, only 4 of 24 GC-LCLs showed significant somatic hypermutation activity, demonstrating that EBV usually silences hypermutation upon infection of GC B cells. Notably, one of three N-LCL showed a low level of intraclonal diversification. Thus, EBV-infection deregulates multiple differentiation factors and processes in B cells, leading to a largely homogenous phenotype of EBV infected B cells in latency III. PMID- 18430473 TI - The effects of cutting solutions on the viability of GABAergic interneurons in cerebral cortical slices of adult mice. AB - To obtain viable GABAergic interneurons in cerebral cortical slices of adult mice, we investigated the effects of slice cutting solutions on the viability of green fluorescent protein (GFP)-expressing cortical neurons in GAD67-GFP knock-in mice. Almost no nuclei of GFP-positive neurons were labeled with propidium iodide in incubated slices, suggesting that GFP fluorescence was a useful indicator for the viability of GABAergic interneurons. When several cutting solutions were compared with saline-based solution, N-methyl-d-glucamine-based sodium-free solution was most effective to keep the number of GFP-positive neurons near the level of perfusion-fixed brain. GFP-positive neurons in slices cut with sodium free solution were more numerous in cortical layers V-VI, at 30-60 microm depth from the cut surface and 1-6h after cutting than those with saline-based solution. Furthermore, the number of GFP-positive neurons decreased in the cutting condition of high calcium concentration (5mM) or high temperature (37 degrees C), and GFP fluorescence decreased when cut at 0 degrees C. The present results indicate that cutting the brain at 20 degrees C in sodium-free solution is a method for preparing cortical slices with GABAergic interneurons viable. This method would thus be useful for electrophysiological and morphological studies of cortical interneurons in slice preparations of the adult brain. PMID- 18430474 TI - Serum markers support disease-specific glial pathology in major depression. AB - BACKGROUND: Recently, it was shown by histopathological studies that mood disorders are characterized by disease-specific glial pathology. METHODS: To validate this hypothesis in vivo we measured weekly and simultaneously serum levels of the neuronal marker neuron-specific enolase and S100B, a protein expressed in astro- and oligodendroglia in the human brain, in 10 patients with major depressive disorder and 10 age- and gender-matched control subjects. Furthermore, we conducted a systematic, quantitative meta-analysis of all published studies on S100B involving 193 patients suffering from mood disorders and 132 healthy control subjects by calculating effect sizes. RESULTS: S100B was elevated at admission and discharge in our patients with major depression compared with control subjects, whereas there were no significant differences for neuron-specific enolase. During treatment S100B decreased slightly, although this effect was not significant. It had no significant impact on neuron-specific enolase. The meta-analysis revealed that serum levels of S100B are consistently elevated in mood disorders during acute major depressive or manic episodes. Additionally, it demonstrated that serum S100B decreases during antidepressive treatment reliably if clinical improvement is sufficient. LIMITATIONS: As the study measured only serum S100B, future (cell culture) studies have to elucidate molecular mechanisms of this protein in mood disorders. Moreover, results have to be replicated in a larger patient group. CONCLUSIONS: S100B may represent a biomarker for mood disorders, particularly major depression, and their treatment. Together with unaltered levels of neuron-specific enolase, our results support in vivo the histopathologically generated hypothesis of disease-specific glial pathology in mood disorders. PMID- 18430470 TI - Neurotoxic profiles of HIV, psychostimulant drugs of abuse, and their concerted effect on the brain: current status of dopamine system vulnerability in NeuroAIDS. AB - There are roughly 30-40 million HIV-infected individuals in the world as of December 2007, and drug abuse directly contributes to one-third of all HIV infections in the United States. Antiretroviral therapy has increased the lifespan of HIV-seropositives, but CNS function often remains diminished, effectively decreasing quality of life. A modest proportion may develop HIV associated dementia, the severity and progression of which is increased with drug abuse. HIV and drugs of abuse in the CNS target subcortical brain structures and DA systems in particular. This toxicity is mediated by a number of neurotoxic mechanisms, including but not limited to, aberrant immune response and oxidative stress. Therefore, novel therapeutic strategies must be developed that can address a wide variety of disparate neurotoxic mechanisms and apoptotic cascades. This paper reviews the research pertaining to the where, what, and how of HIV and cocaine/methamphetamine toxicity in the CNS. Specifically, where these toxins most affect the brain, what aspects of the virus are neurotoxic, and how these toxins mediate neurotoxicity. PMID- 18430475 TI - An epidemiological survey of alcohol use disorders in a Tibetan population. AB - We performed an epidemiological survey in order to detect the prevalence of alcohol use disorders in a sub-group of the population of Tibet. The Alcohol Use Disorders Identification Test (AUDIT) questionnaire, the Severity of Alcohol Dependence Questionnaire (SADQ), and a 12-item version of the General Health Questionnaire (GHQ12) were used to obtain epidemiological data on alcohol use disorders and to assess the severity of 'problem drinking' and general mental health status. The AUDIT is a reliable and valid screening tool for both alcohol abuse and dependence in the Tibetan population to identify individuals with alcohol use problems. The cut-off points were set to be 10 and 13 of the AUDIT scores as a diagnostic discriminator of alcohol abuse and alcohol dependence, respectively, with both sensitivity and specificity>0.84. The prevalence of alcohol abuse, was 2.7% (female: 2.0%; male: 6.2%), alcohol dependence 13.5% (female: 7.6%; male: 25.4%) and alcohol use disorders 16.2% (female: 9.6%; male: 31.6%). Age and sex were the main factors affecting an individual's alcohol use and general mental health status. The epidemiological data on alcohol use disorders documented in this project may be helpful in future work seeking more valid causal inferences or interpretations related to this prevalent health problem in Tibet. PMID- 18430476 TI - Impaired instrumental choice in crib-biting horses (Equus caballus). AB - Horses displaying an oral stereotypy were tested on an instrumental choice paradigm to examine differences in learning from non-stereotypic counterparts. Stereotypic horses are known to have dysfunction of the dorsomedial striatum, and lesion studies have shown that this region may mediate response-outcome learning. The paradigm was specifically applied in order to examine learning that requires maintenance of response-outcome judgements. The non-stereotypic horses learned, over three sessions, to choose a more immediate reinforcer, whereas the stereotypic horses failed to do so. This suggests an initial behavioural correlate for dorsomedial striatum dysregulation in the stereotypy phenotype. PMID- 18430477 TI - Expression of metallothionein mRNAs by in situ hybridization in the gills of Mytilus galloprovincialis, from natural polluted environments. AB - Metallothioneins (MTs), metal-inducible proteins, are crucial proteins for the regulation of essential metals, and are transcriptionally induced in all organisms by certain heavy metals, oxidative stress and inflammation. The gills represent an organ of uptake and loss of metals in which different mechanisms are present controlling the functions directly involved in the maintenance of homeostasis. In this study, the morphological and histomorphological aspects of branchial epithelium in Mytilus galloprovincialis from polluted environment (Faro swamp, Messina, Italy) have been investigated. The reverse transcriptase polymerase chain reaction (PCR) has been used to isolate complementary DNA of both MT isoforms present from RNA extracted from mussel gills. The respective mRNAs on histological sections have been visualized by in situ hybridization. These methods showed that MT-10 mRNA is expressed at the basal level. In contrast, the MT-20 expression level was very low under basal conditions, while its mRNA increased dramatically in individuals collected in Faro. The presence of acid mucocytes and MTs in the gills may be considered a further defensive mechanism also related to the significantly higher concentration of Cd, Pb and Cr found in gills of M. galloprovincialis from Faro than specimens from the reference site (Goro). The results obtained show that, in stressed mussels, the defensive processes increase to maintain the normal functions of the organs more exposed to the action of polluted substances. PMID- 18430478 TI - Quantitative risk assessment of foot-and-mouth disease introduction into Spain via importation of live animals. AB - Spain has been a foot-and-mouth disease (FMD)-free country since 1986. However, the FMD epidemics that recently affected several European Union (EU) member countries demonstrated that the continent is still at high risk for FMD virus (FMDV) introduction, and that the potential consequences of those epidemics are socially and financially devastating. This paper presents a quantitative assessment of the risk of FMDV introduction into Spain. Results suggest that provinces in north-eastern Spain are at higher risk for FMDV introduction, that an FMD epidemic in Spain is more likely to occur via the import of pigs than through the import of cattle, sheep, or goats, and that a sixfold increase in the proportion of premises that quarantine pigs prior to their introduction into the operation will reduce the probability of FMDV introduction via import of live pigs into Spain by 50%. Allocation of resources towards surveillance activities in regions and types of operations at high risk for FMDV introduction and into the development of policies to promote quarantine and other biosecurity activities in susceptible operations will decrease the probability of FMD introduction into the country and will strengthen the chances of success of the Spanish FMD prevention program. PMID- 18430479 TI - Enzyme catalysis on solid surfaces. AB - Enzyme-catalysed reactions in which substrates are bound (immobilised) to solid surfaces are becoming increasingly important in biotechnology. There is a general drive for miniaturisation and automation in chemistry and biology, and immobilisation of the reaction intermediates and substrates, for example on microarrays or nanoparticles, helps to address technical challenges in this area. In bionanotechnology, enzyme catalysis can provide highly selective and biocompatible tools for the modification of surfaces on the nano-scale. Here, we review the range of enzyme-catalysed reactions that have been successfully performed on the solid phase and discuss their application in biotechnology. PMID- 18430480 TI - Artificial molecular sieves and filters: a new paradigm for biomolecule separation. AB - Patterned regular sieves and filters with comparable molecular dimensions hold great promise as an alternative to conventional polymeric gels and fibrous membranes to improve biomolecule separation. Recent developments of microfabricated nanofluidic sieves and filters have demonstrated superior performance for both analytical and preparative separation of various physiologically relevant macromolecules, including proteins. The insights gained from designing these artificial molecular sieves and filters, along with the promising results gathered from their first applications, serve to illustrate the impact that they can have on improving future separation of complex biological samples. Further development of artificial sieves and filters with more elaborate geometrical constraints and tailored surface functionality is believed to provide more promising ideals and results for biomolecule separation, which has great implications for proteomic research and biomarker discovery. PMID- 18430481 TI - Alpha power and coherence primarily reflect neural activity related to stages of motor response during a continuous monitoring task. AB - Previously, EEG theta (4-6 Hz) was related to goal conflict resolution [Moore, R.A., Gale, A., Morris, P.H., Forrester, D., 2006. Theta phase locking across the neocortex reflects cortico-hippocampal recursive communication during goal conflict resolution. Int. J. Psychophysiol. 60, 260-273] in the context of theory linked with animal hippocampal theta [Gray, J.A., McNaughton, N., 2000. The Neuropsychology of Anxiety: An Enquiry into the Functions of the Septo Hippocampal system, 2nd ed, Oxford University Press, Oxford]. Here, the hypothesis that human EEG alpha (8-12 Hz) may also be a natural analogue to animal hippocampal theta is tested. Participants engaged in a monitoring task where the object was to press a response key immediately after presentation of 4 individual, non-repeating, single integer odd digits. These were presented amongst a continuous stream of single integer digits and Xs. EEG recorded in the earlier study were reanalysed; this time extracting alpha power and coherence from the same 34 participants. Alpha had a different profile to theta and was not primarily related to goal conflict. Low alpha (8-10 Hz) coherence consistently increased at electrodes close to primary sensorimotor cortex; particularly during response execution and response inhibition. The coherence analysis revealed that high alpha (10-12 Hz) related to response execution. Supplementary analyses demonstrated widespread high alpha coherence increase during response execution, inhibition and preparation. These data were discussed within the context of motor driven 'classic alpha' and Rolandic mu. A coherence profile which differentiated response execution and response inhibition was proposed to reflect a working memory network which was activated during response execution. Also, alpha power (8-12 Hz) reduced at several central electrodes during response execution. This reflected classic Rolandic mu response. Participants displaying a predicted low alpha power trend had the fastest response times; this was linked with traditional views of low alpha's functional significance. PMID- 18430482 TI - Ultrafast pyrosequencing of Corynebacterium kroppenstedtii DSM44385 revealed insights into the physiology of a lipophilic corynebacterium that lacks mycolic acids. AB - Corynebacterium kroppenstedtii is a lipophilic corynebacterial species that lacks in the cell envelope the characteristic alpha-alkyl-beta-hydroxy long-chain fatty acids, designated mycolic acids. We report here the bioinformatic analysis of genome data obtained by pyrosequencing of the type strain C. kroppenstedtii DSM44385 that was initially isolated from human sputum. A single run with the Genome Sequencer FLX system revealed 560,248 shotgun reads with 110,018,974 detected bases that were assembled into a contiguous genomic sequence with a total size of 2,446,804bp. Automatic annotation of the complete genome sequence resulted in the prediction of 2122 coding sequences, of which 29% were considered as specific for C. kroppenstedtii when compared with predicted proteins from hitherto sequenced pathogenic corynebacteria. This comparative content analysis of the genome data revealed a large repertoire of genes involved in sugar uptake and central carbohydrate metabolism and the presence of the mevalonate route for isoprenoid biosynthesis. The lack of mycolic acids and the lipophilic lifestyle of C. kroppenstedtii are apparently caused by gene loss, including a condensase gene cluster, a mycolate reductase gene, and a microbial type I fatty acid synthase gene. A complete beta-oxidation pathway involved in the degradation of fatty acids is present in the genome. Evaluation of the genomic data indicated that lipophilism is the dominant feature involved in pathogenicity of C. kroppenstedtii. PMID- 18430483 TI - Combinatorial biosynthesis of 5-O-desosaminyl erythronolide A as a potent precursor of ketolide antibiotics. AB - Ketolides, characterized by possessing a 3-keto group in place of the l-cladinose moiety of erythromycin A, are the recent generation of antimicrobials derived semi-synthetically from the 14-membered ring macrolide erythromycin A. The multi step synthetic route to ketolides can be shortened by using 5-O-desosaminyl erythronolide A as a precursor, which reduces the steps for the removal of l cladinose attached at the C-3 position in erythromycin A. Deletion of an eryBV gene encoding mycarosyl glycosyltransferase in the erythromycin-producer Saccharopolyspora erythraea resulted in the accumulation of 5-O-desosaminyl erythronolide B. In vivo expression of the cytochrome P450 gene pikC, which encodes the substrate-flexible hydroxylase from the pikromycin biosynthetic pathway of Streptomyces venezuelae, in the eryBV deletion mutant strain of Sac. erythraea led to 5-O-desosaminyl erythronolide A production. PMID- 18430484 TI - Recombinant L and P protein complex of Rinderpest virus catalyses mRNA synthesis in vitro. AB - Rinderpest virus belongs to the family of Paramyxoviridae, consisting of non segmented negative sense RNA viruses. Viral transcription and replication are carried out by the RNA dependent RNA polymerase L protein which functions together with P protein as L-P complex. The exact events triggering the polymerase complex from transcription to replication function is poorly understood. In the present work, an in vitro transcription system has been described with partially purified L-P complex expressed in insect cells and viral genomic RNA. The relative abundance of each species of mRNA synthesized in vitro decreased from the 3' end of the genome to the 5' end similar to their abundance in virus infected cells. Recombinant L-P complex was unable to synthesize leader RNA suggesting the initiation of transcription from gene start site and not at the 3' end of the genome. PMID- 18430485 TI - Building a liver disease policy in the European Union. PMID- 18430486 TI - Influence of blood transfusions and preoperative anemia on long-term survival in patients operated for non-small cell lung cancer. AB - It has been postulated that transfusions have immunosuppressive effects that promote tumor growth and metastasis. Moreover perioperative anemia is considered an independent prognostic factor on outcome in patients operated for malignancy. We evaluated the influence of red blood cell (RBC) transfusions and perioperative anemia on survival in non-small cell lung carcinoma (NSCLC) patients. From 1999 through 2005, 331 consecutive patients, male/female=295/36 (mean age 64+/-9 years), who underwent radical surgery for NSCLC were prospectively enrolled in this cohort and followed up for a mean of 27.2 months. The overall survival of patients was analyzed in relation to RBC transfusions and perioperative anemia. These parameters were analyzed in the whole cohort of patients and separately for stage I patients. Patients were divided according to perioperative transfusion, into Group A (transfused) and Group B (non-transfused) and according to the preoperative haemoglobin (Hb) level into Group 1(Hb<12g/dl) and Group 2(Hb> or =12g/dl), respectively. The overall transfusion rate was 25.7%. Univariate analysis showed that in the whole cohort of patients overall survival was significantly shorter in Group A (mean 33.6 months, 5-year survival 25.1%) compared to Group B (mean 48.0 months, 5-year survival 37.3%) (p=0.001). It also showed that patients with preoperative Hb level <12g/dl (Group 1), (mean of 33.0 months, 5-year survival 21.3%) had shorter survival compared to Group 2 patients (mean 49.3 months and 5-year survival 40.0%), respectively (p=0.002). Multivariate analysis in the whole cohort of patients showed that preoperative anemia was an independent risk factor for survival while RBC transfusion was not. In particular for stage I patients, it was shown that RBC transfusion was an independent prognostic factor for long-term survival as detected by multivariate analysis (p=0.043), while anemia was not. RBC transfusions affect adversely the survival of stage I NSCLC patients, while do not exert any effect on survival of patients with surgically resectable more advanced disease, where preoperative anemia is an independent negative prognostic factor. These findings indicate that RBC transfusion might exert an immunomodulatory effect on patients with early disease while in more advanced stages this effect is not apparent. PMID- 18430487 TI - Regulation of photosynthesis by sugars in sugarcane leaves. AB - In sugarcane, increased sink demand has previously been shown to result in increased photosynthetic rates that are correlated with a reduction in leaf hexose concentrations. To establish whether sink limitation of photosynthesis is a result of sugar accumulation in the leaf, excision and cold-girdling techniques were used to modify leaf sugar concentrations in pot-grown sugarcane. In excised leaves that were preincubated in darkness for 3h, sucrose accumulation was reduced but accumulated again upon transfer to the light, while hexose concentrations remained lower than in controls (7.7 micromol mg(-1)FW versus 18.6 micromol mg(-1)FW hexose in controls). These results were associated with a 66% and 59% increase in photosynthetic assimilation (A) and electron transport rate (ETR), respectively, compared to controls maintained in the light. Similar increases in photosynthesis were observed when dark-treated leaves were supplied with 5mM sorbitol, but not when supplied with 5mM sucrose. Further analyses of (14)C-labeled sugars indicated rapid turnover between sucrose and hexose. Cold girdling (5 degrees C) increased sucrose and hexose levels and resulted in a decline of photosynthetic rates over 5d (48% and 35% decline in assimilation rate and ETR, respectively). These sugar-induced changes in photosynthesis were independent of changes in stomatal conductance. This study demonstrates that the down-regulation of photosynthesis in response to culm sugar accumulation reported previously could be due to the knock-on effect of accumulation of sugar in leaf tissue, and supports the contention that hexose, rather than sucrose, is responsible for the modulation of photosynthetic activity. PMID- 18430488 TI - The proliferation of amyloplasts in meristematic cells of developing stolons of potato and apple callus: progenitors of proplastids. AB - To monitor the events in the proliferation of amyloplasts, the ultrastructure of relevant structures in the cytosol has to be studied. For this investigation, photographs of cellular ultrastructures in developing potato stolons and apple callus were taken and examined. The images indicated that the contribution to proliferation of the division of mature amyloplasts was extremely low and that the major pathway involved the generation of the proplastids from "mother" amyloplasts. The generation of proplastids was followed either by division into small bodies of 1microm or less in diameter or by growth to slender proplastids of 5microm in length. The elongated proplastids multiplied by splitting at random sites, with subsequent enlargement to mature sizes. The latter process contributed to the massive accumulation of amyloplasts in cells but has not previously been adequately emphasized. With respect to the putative "mother" amyloplasts, numerous divergent amyloplasts were observed with a considerably different ultrastructure compared to the normal types, and with a characteristically extended and constricted stroma. Various lines of evidence indicated that the divergent amyloplasts were the "mother" amyloplasts of the proplastids. No other plastidic organelles with features that suggest the generation of proplastids were detected in the cytosol. PMID- 18430489 TI - Investigation into the mechanism of stimulation by low-concentration stressors in barley seedlings. AB - Beneficial effects of low-concentration chemical stressors have been investigated previously in different model systems. The symptoms of stimulation are known from earlier studies, but information about the mechanism is at an initial stage. In the present work, the mechanism of stimulation of low-concentration Cd (5 x 10( 8)M) and 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU, 10(-7)M) was investigated in barley seedlings. In treated plants, the amount of cytokinins increased in roots and, after being transported to the leaves, they caused stimulation there. To identify the signal transduction pathway(s) involved in the primary stimulation of cytokinin synthesis (and/or activation) in roots, specific phosphatidylinositol-4,5-bisphosphate-inositol-1,4,5-triphosphate/diacylglycerol (PIP(2)-IP(3)/DAG) and mitogen activated protein kinase (MAPK) signaling pathway inhibitors were added to the nutrient solution, and all proved to be effective, eliminating the stimulation by the stressors. Measurements of superoxide dismutase (SOD, EC 1.15.1.1) activity and the amount of malonyldialdehyde (MDA) showed that the increased amount of Cd did not cause oxidative stress in the roots, and no oxidative stress was found in the leaves, where Cd did not even accumulate. DCMU slightly increased the activity of SOD after 1 week in roots, but did not cause lipid peroxidation. In leaves, there was no oxidative stress upon treatment with DCMU. Thus, oxidative stress cannot be responsible for the stimulation with low-concentration stressors, as they changed the activity of SOD differently, while being equally stimulative for the plants. PMID- 18430490 TI - Beliefs about fast food in Australia: a qualitative analysis. AB - The consumption of energy-dense fast foods has been implicated as a causal factor in the development of obesity. The development of strategies to modify food choice behaviour requires an understanding of the behaviour and the driving factors. This study examined the rationale behind decisions to either choose or avoid fast foods. Drawing partly on the Theory of Planned Behaviour (Ajzen, (1988)), a qualitative design was employed to examine the beliefs and perceptions associated with fast-food consumption within an Australian sample. Findings provided an indication that positive affective reactions to fast food, convenience, and self-serving cognitions may override cognitive analyses of the longer-term health risks associated with frequent fast-food consumption. PMID- 18430491 TI - BMAP-28 improves the efficacy of vancomycin in rat models of gram-positive cocci ureteral stent infection. AB - An experimental study was performed to evaluate the efficacy of BMAP-28 alone and in combination with vancomycin in animal models ureteral stent infection due to Enterococcus faecalis and Staphylococcus aureus. Study included a control group without bacterial challenge to evaluate the sterility of surgical procedure, a challenged control group that did not receive any antibiotic prophylaxis and for each bacterial strain three challenged groups that received (a) 10 mg/kg vancomycin intraperitoneally, immediately after stent implantation, (b) BMAP-28 coated ureteral stents where 0.2-cm(2) sterile ureteral stents were incubated in 1mg/l BMAP-28 solution for 30 min immediately before implantation and (c) intraperitoneal vancomycin plus BMAP-28-coated ureteral stent at the above concentrations. Experiments were performed in duplicate. Ureteral stents were explanted at day 5 following implantation and biofilm bacteria enumerated. Our data showed that rats that received intraperitoneal vancomycin showed the lowest bacterial numbers. BMAP-28 combined with vancomycin showed efficacies higher than that of each single compound. These results highlight the potential usefulness of this combination in preventing ureteral stent-associated in gram-positive infections. PMID- 18430492 TI - Penetratin and derivatives acting as antifungal agents. AB - The synthesis, in vitro evaluation, and conformational study of RQIKIWFQNRRMKWKK NH(2) (penetratin) and related derivatives acting as antifungal agents are reported. Penetratin and some of its derivatives displayed antifungal activity against the human opportunistic pathogenic standardized ATCC strains Candida albicans and Cryptococcus neoformans as well as clinical isolates of C. neoformans. Among the compounds tested, penetratin along with the nonapeptide RKWRRKWKK-NH(2) and the tetrapeptide RQKK-NH(2) exhibited significant antifungal activities against the Cryptococcus species. A comprehensive conformational analysis on the peptides reported here using three different approaches, molecular mechanics, simulated annealing and molecular dynamics simulations, was carried out. The experimental and theoretical results allow us to identify a topographical template which may provide a guide for the design of new compounds with antifungal characteristics against C. neoformans. PMID- 18430493 TI - Co-administration of polyphosphazenes with CpG oligodeoxynucleotides strongly enhances immune responses in mice immunized with Hepatitis B virus surface antigen. AB - An emerging paradigm in vaccinology is that multiple adjuvant combinations may be more effective than individual adjuvants in enhancing immune responses to vaccine antigens. We investigated whether the polyphosphazenes used in combination with CpG oligodeoxynucleotides (ODN) were potent adjuvant formulations. BALB/c mice were immunized subcutaneously with Hepatitis B surface antigen (HBsAg) alone, or in various combinations with poly[di(sodium carboxylatophenoxy)phosphazene] (PCPP), poly[di(sodium carboxylatoethylphenoxy)phosphazene] (PCEP) or CpG ODN. All three adjuvants enhanced HBsAg-specific IgG1 antibody responses with PCEP inducing the highest responses. PCEP and CpG ODN significantly enhanced the Th1 associated antibody isotype IgG2a. As expected CpG ODN induced predominantly Th1 type immune responses while PCEP was associated with mixed Th1/Th2 immune responses. Interestingly, PCEP and PCPP synergized with CpG ODN to further enhance antibody responses. Since the mechanisms which mediate the adjuvant activity of polyphosphazenes are not fully understood, we investigated whether PCEP and PCPP could stimulate innate immune responses. Incubation of mouse splenocytes with PCEP or PCPP (in the absence of antigen) stimulated production of IL-4 and IL-12, but only PCEP induced significant IFNgamma production. Additionally, IL-12 was not required for PCEP induced IFNgamma response. We conclude that the polyphosphazene-CpG ODN combination is a potent adjuvant formulation that is more effective in enhancing immune responses than either of the individual adjuvants. In addition, we provide evidence that PCEP and PCPP can stimulate innate cytokine production, suggesting a potential mechanism by which polyphosphazenes achieve their potent adjuvant effects. PMID- 18430494 TI - Evaluation of ISCOM-adjuvanted subunit vaccines containing recombinant feline immunodeficiency virus Rev, OrfA and envelope protein in cats. AB - For the development of feline immunodeficiency virus (FIV) vaccines mostly structural proteins have been evaluated for their capacity to induce protective immunity. In the present study, subunit vaccines containing recombinant FIV accessory proteins Rev and OrfA were evaluated in cats. Cats were vaccinated repeatedly with these proteins, adjuvanted with immune stimulating complexes (ISCOMs). In addition, cats were vaccinated with bacterially expressed fragments spanning the entire FIV envelope protein, either alone or in combination with the regulatory proteins. Subsequently, the cats were challenged with a homologous FIV strain to assess the level of protective immunity achieved with the respective vaccination regimens. Although the vaccines proved to be immunogenic, vaccinated cats were not protected from infection with FIV. PMID- 18430495 TI - Combined effects of IL-12 and electroporation enhances the potency of DNA vaccination in macaques. AB - DNA vaccines are a promising technology. Historically, however, the ability of DNA vaccines to induce high response rates and strong immune responses, especially antibody responses, in non-human primates and human clinical trials has proven suboptimal. Here, we performed a pilot study in rhesus macaques to evaluate whether we could improve the immunogenicity of DNA vaccines through the use of adjuvant technology and improved delivery systems. The study consisted of four groups of animals that received: DNA by intramuscular (IM) injection, DNA with plasmid-encoded IL-12 by IM injection, DNA by IM injection with in vivo electroporation (EP), and DNA with IL-12 by IM EP. Each group was immunized three times with optimized HIV gag and env constructs. Vaccine immunogenicity was assessed by IFNgamma ELISpot, CFSE proliferation, polyfunctional flow cytometry, and antibody ELISA. Similar to previous studies, use of IL-12 as an adjuvant increased the gag and env-specific cellular responses. The use of EP to enhance plasmid delivery resulted in dramatically higher cellular as well as humoral responses. Interestingly, the use of EP to administer the DNA and IL-12 adjuvant combination resulted in the induction of higher, more efficient responses such that a 10-fold increase in antigen-specific IFNgamma(+) cells compared to IM DNA immunization was observed after a single immunization. In addition to increases in the magnitude of IFNgamma production in the initial and memory responses, the combined approach resulted in enhancements in the proliferative capacity of antigen-specific CD8(+) T cells and the amount of polyfunctional cells capable of producing IL-2 and TNFalpha in addition to IFNgamma. These data suggest that adjuvant and improved delivery methods may be able to overcome previous immunogenicity limitations in DNA vaccine technology. PMID- 18430496 TI - The Intensive Vaccines Monitoring Programme (IVMP): an electronic system to monitor vaccine safety in New Zealand. AB - New Zealand introduced a tailor-made vaccine (MeNZB) for epidemic control of Group B meningococcal disease. The Intensives Vaccine Monitoring Programme (IVMP), which prospectively collected data electronically on a cohort of children receiving vaccinations in sentinel practices across NZ, was developed as part of a national multi-faceted safety strategy. The main aim of the IVMP was to identify the presence of unexpected adverse events occurring with MeNZB vaccination. We describe the methodology and success factors plus consider the limitations encountered in this system which shows potential as a means for post marketing vaccine and medicine surveillance in the future. PMID- 18430497 TI - Radium removal from mine waters in underground treatment installations. AB - The underground mining of hard coal is widespread in the Upper Silesian Coal Basin (southern Poland). In deep mines, inflows of highly mineralised waters containing radium isotopes are numerous. These waters cause severe damage to the natural environment due to the salinity, but additionally radioactive pollution occurs. The region is densely populated, therefore mitigation methods are very important. The method of radium removal has been applied in full technical scale in two coal mines with very good results--in one of the mines radium-bearing waters are treated at the rate of approximately 0.1 m3 s(-1), while in another mine salty waters are purified at the rate of 0.1 m3 s(-1). The purification takes place in special underground galleries without any contact of the mining crew with the radioactive deposits produced during the process. As a result, release of radium is significantly lower, more than 200 MBq of 226Ra and 228Ra remains underground each day. PMID- 18430498 TI - Current approach to hemochromatosis. AB - Iron overload diseases of genetic origin are an ever changing world, due to major advances in genetics and molecular biology. Five major categories are now established: HFE-related or type1 hemochromatosis, frequently found in Caucasians, and four rarer diseases which are type 2 (A and B) hemochromatosis (juvenile hemochromatosis), type 3 hemochromatosis (transferrin receptor 2 hemochromatosis), type 4 (A and B) hemochromatosis (ferroportin disease), and a(hypo)ceruloplasminemia. Increased duodenal iron absorption and enhanced macrophagic iron recycling, both due to an impairment of hepcidin synthesis, account for the development of cellular excess in types 1, 2, 3, and 4B hemochromatosis whereas decreased cellular iron egress is involved in the main form of type 4A) hemochromatosis and in aceruloplasminemia. Non-transferrin bound iron plays an important role in cellular iron excess and damage. The combination of magnetic resonance imaging (for diagnosing visceral iron overload) and of genetic testing has drastically reduced the need for liver biopsy. Phlebotomies remain an essential therapeutic tool but the improved understanding of the intimate mechanisms underlying these diseases paves the road for innovative therapeutic approaches. PMID- 18430499 TI - Reduced mercury deposition in New Hampshire from 1996 to 2002 due to changes in local sources. AB - Changes in deposition of gaseous divalent mercury (Hg(II)) and particulate mercury (Hg(p)) in New Hampshire due to changes in local sources from 1996 to 2002 were assessed using the Industrial Source Complex Short Term (ISCST3) model (regional and global sources and Hg atmospheric reactions were not considered). Mercury (Hg) emissions in New Hampshire and adjacent areas decreased significantly (from 1540 to 880 kg yr(-1)) during this period, and the average annual modeled deposition of total Hg also declined from 17 to 7.0 microg m(-2) yr(-1) for the same period. In 2002, the maximum amount of Hg deposition was modeled to be in southern New Hampshire, while for 1996 the maximum deposition occurred farther north and east. The ISCST3 was also used to evaluate two future scenarios. The average percent difference in deposition across all cells was 5% for the 50% reduction scenario and 9% for the 90% reduction scenario. PMID- 18430500 TI - Variation in serum and plasma PTH levels in second-generation assays in hemodialysis patients: a cross-sectional study. AB - BACKGROUND: Previous reports show that parathyroid hormone (PTH) concentrations may vary widely depending on the assay used to assess PTH. In this cross sectional study, we aim to determine the usefulness of standardizing blood handling for optimal interpretation of PTH in patients with chronic kidney disease. STUDY DESIGN: Diagnostic test study. SETTING & PARTICIPANTS: Predialysis blood was sampled in 34 long-term hemodialysis patients at a single academic medical center. INDEX TEST: PTH was measured by using 6 different automated second-generation assays (Elecsys, Advia Centaur, LIAISON, Immulite, Architect, and Access assays), 3 blood specimen types (serum, EDTA plasma, and citrate plasma), and 2 consecutive days of measurement (after thawing and 18 hours later with samples having been let at room temperature). REFERENCE TEST: None. RESULTS: A mixed statistical analysis model showed that the nature of the assay (P < 0.001) and nature of the blood sample (P < 0.001) significantly influenced variability in PTH concentrations, whereas day of measurement (day 1 or 2) did not (P = 0.5). Most PTH variability was caused by observations (96.8%), then manufacturer's kit (2.5%), and last, specimen type (0.7%). PTH concentrations measured in citrate plasma were lower with every assay method used than those observed in serum or EDTA plasma. The interaction between manufacturer and specimen type was of moderate statistical significance (P = 0.04). To evaluate the potential clinical consequence of PTH measure variability, we classified patients according to Kidney Disease Outcomes Quality Initiative cutoff values (PTH < 150 pg/mL; PTH, 150 to 300 pg/mL; and PTH > 300 pg/mL). Overall, statistical classification agreement was moderate to high for comparison between assays and high to very high between different blood samples and between days of measurement. However, we found that up to 11 of 34 patients were classified in different categories with some assays (LIAISON versus Architect) and up to 7 of 34 in different categories with different blood specimen type (citrate plasma versus serum [corrected] in LIAISON assay). LIMITATIONS: This is a cross sectional study that used single lots of reagents. There currently is no reference method for the measurement of PTH and no recombinant PTH standard for PTH assay. CONCLUSION: PTH variability caused by the nature of the assay and/or blood specimen type is large enough to potentially influence clinical decision making. A specified collection method therefore should be used for PTH measurements. In routine practice, we recommend serum PTH over EDTA or citrate plasma. PMID- 18430501 TI - Intimate partner violence theoretical considerations: moving towards a contextual framework. AB - Several theories have been developed to provide a conceptual understanding of intimate partner violence (IPV) episodes. Although each of these theories has found some degree of empirical support, they are limited in their explanatory power of IPV episodes and their ability to significantly impact the efficacy of IPV prevention and treatment programs. The current paper provides a review and critique of current IPV theories and highlights strategies for improving upon these theories. An alternative theoretical conceptualization is introduced that incorporates existing IPV and functional analytic literature into a contextual framework for conceptualizing IPV episodes. Components of the IPV contextual framework include distal, static and proximal antecedents; motivating factors; behavioral repertoire; discriminative stimuli (i.e. environmental cues/signals); verbal rules; and IPV consequences. The proposed theoretical framework offers two primary advantages over former IPV theories. First, it provides a comprehensive conceptualization of IPV by integrating components of previous IPV theories and their related empirical findings into one, cohesive conceptual framework. Additionally, it allows for a more fine-grained analysis of more proximal variables potentially related to discrete IPV episodes. A discussion of how the proposed theoretical framework may influence future IPV research and clinical practice is provided. PMID- 18430502 TI - State-dependent changes in the expression levels of NCAM-140 and L1 in the peripheral blood cells of bipolar disorders, but not in the major depressive disorders. AB - Recent postmortem brain and imaging studies provide evidence for disturbances of structural and synaptic plasticity in patients with mood disorders. Several lines of evidence suggest that the cell adhesion molecules (CAMs), neural cell adhesion molecules (NCAM) and L1, play important roles in both structural and synaptic plasticity. Although postmortem brain studies have indicated altered expression levels of NCAM and L1, it is still unclear whether these changes are state- or trait-dependent. In this study, the mRNA levels for various CAMs, including NCAM and L1, were measured using quantitative real-time PCR in peripheral blood cells of major depressive disorder patients, bipolar disorder patients and normal healthy subjects. Reduced expression levels of NCAM-140 mRNA were observed in bipolar disorder patients in a current depressive state. In contrast, L1 mRNA levels were increased in bipolar disorder patients in a current depressive state. NCAM-140 and L1 mRNA levels were not changed in bipolar disorder patients in a remissive state, or in major depressive disorder patients. In addition, there were no significant changes in the expression levels of intercellular adhesion molecule -1, vascular cell adhesion molecule -1, E-cadherin, or integrin alphaD among healthy controls, major depressive or bipolar disorder patients. Our results suggest that the reciprocal alteration in the expression of NCAM-140 and L1 mRNAs could be state-dependent and associated with the pathophysiology of bipolar disorder. PMID- 18430503 TI - Family-based association studies of CAPON and schizophrenia in the Chinese Han population. AB - Although there is evidence pointing to CAPON as a susceptible gene for schizophrenia, the results of independent association studies have so far been inconsistent. A recent case-control study by Zheng et al. supported CAPON as a susceptible site for the disease in the Chinese Han population. In their study both the single polymorphism (rs348624) and individual haplotypes showed significant association with schizophrenia. Our study further investigates this relationship this time using a family-based association. We selected 5 SNPs including rs348624 and performed a Transmission Disequilibrium Test (TDT) in 319 Chinese Han trios. Our results identified no single marker nor haplotype associated with schizophrenia, which did not suggest that CAPON was a susceptible site in the Chinese Han population, or it appeared unlikely that the CAPON played a major role in the aetiology of schizophrenia. Since there is consistent evidence pointing to 1q21-22 as a positional candidate region for schizophrenia, we suggest that further research should focus on other genes located in this region. PMID- 18430504 TI - Fungal contamination and Aflatoxin B1 and Ochratoxin A in Lebanese wine-grapes and musts. AB - Five hundred and ten strains of filamentous fungi were isolated from Lebanese grapes during 2005 at veraison and harvesting periods. Four hundred eighty-seven isolates belonged to the Aspergillus spp. (95.5%) and 23 belonged to the Penicillium spp. (4.5%). Black aspergilli constituted 56.9% (52.2% Aspergillus niger aggregates, 2.9% Aspergillus japonicus and 1.8% Aspergillus carbonarius) while the isolation rate of Aspergillus flavus the none habitual member of grape mycobiota was 43.1% of the total Aspergillus spp. isolated. All isolates were tested for the ability to produce the Ochratoxin A (OTA) and the Aflatoxin B1 (AFB1). A. carbonarius showed that it is the only species able to produce the OTA with a production ability of 100% and a maximum concentration reaching 8.38microg/g CYA. As for the aflatoxigenic ability, 43.4% of A. flavus isolates produced this mycotoxin with a maximum production reaching 22.6microg/g CYA while none of the other isolates showed a production capacity of this mycotoxin. Forty seven samples of must produced from the collected grapes were also analyzed. None of these samples was contaminated by OTA at a detectable limit while 40% of these same samples were found to contain AFB1 with concentrations ranging from 0.01 to 0.46microgl(-1). PMID- 18430505 TI - Development and validation of a microemulsion electrokinetic chromatography method for patulin quantification in commercial apple juice. AB - A microemulsion electrokinetic chromatography (MEECK) method for patulin (PAT) quantification in apple juice samples has been developed. The effects of several important factors such as co-surfactant type, concentration of surfactant, acetonitrile percentage in the microemulsion, and running voltage and temperature were investigated to determine the optimum conditions. They resulted to be: a background electrolyte (BGE) composed of 25mM of sodium tetraborate, SDS (2.16%w/w), ethanol (6.49%w/w), n-octanol (0.82%w/w) and 2%v/v acetonitrile; applied voltage of +15kV; and a capillary temperature of 35 degrees C. PAT was detected at 276nm. Quantification and detection limits (LOQ and LOD) in apple juice samples were 8.0microgL(-1) and 3.2microgL(-1), respectively. Patulin was extracted from apple juice using ethyl acetate with a mean recovery value of 75.3% (RSD=4.5). This method was applied to the measurement of patulin in twenty commercial apple juice samples obtained from different Danish supermarkets. The PAT apple juice mean and median levels obtained were 35.9 and 10.9microgL(-1), respectively. The comparison with a previously validated micellar electrokinetic chromatography (MEKC) method for PAT analysis showed the suitability of using MEEKC for this mycotoxin analysis. However, the expectations of obtaining a higher efficiency and thus lower limits of detection and quantitation when using MEEKC were not met. PMID- 18430506 TI - The fracture behavior of premolar teeth with class II cavities restored by both direct composite restorations and endodontic post systems. AB - PURPOSE: The aim of the study was to evaluate the influence of different posts on the fracture load and fracture mode of endodontically treated premolars with class II cavities and direct composite restorations in an ex vivo setting. METHODS: Forty-eight single-rooted human premolars were endodontically treated and prepared with standardized MO (mesio-occlusal) cavities. Eight teeth each received either no posts or were restored with titanium screws (BKS), glass fiber posts (DentinPost), or quartz fiber posts (DT Light SL). Sixteen teeth were restored with zirconium dioxide posts (CeraPost). BKS-screws and eight zirconium dioxide posts were cemented conventionally with glass ionomer cement; Panavia F resin cement was used for all others. The specimens were restored with direct composite restorations. Eight sound premolars served as the controls. After thermomechanical fatigue testing, the samples were loaded until fracture occurred at an angle of 45 degrees . All specimens were evaluated for fracture lines. RESULTS: The sound teeth showed the significant highest fracture load (792.50+/ 210.01 N). The group restored with quartz fiber posts differed not significantly from the control. In the groups with fiber posts and titanium screws significant higher fracture load values occurred as in the group with direct composite restorations without posts. The groups with fiber posts did not show a more favorable fracture mode than the other groups. CLINICAL SIGNIFICANCE: The use of an intraradicular post in premolars with class II cavities can significantly increase the resistance towards extra-axial forces. PMID- 18430507 TI - Adsorption characteristics of Cu and Ni on Irish peat moss. AB - Peat has been widely used as a low cost adsorbent to remove a variety of materials including organic compounds and heavy metals from water. Various functional groups in lignin allow such compounds to bind on active sites of peat. The adsorption of Cu(2+) and Ni(2+) from aqueous solutions on Irish peat moss was studied both as a pure ion and from their binary mixtures under both equilibrium and dynamic conditions in the concentration range of 5-100mg/L. The pH of the solutions containing either Cu(2+) or Ni(2+) was varied over a range of 2-8. The adsorption of Cu(2+) and Ni(+2) on peat was found to be pH dependent. The adsorption data could be fitted to a two-site Langmuir adsorption isotherm and the maximum adsorption capacity of peat was determined to be 17.6 mg/g for Cu(2+) and 14.5mg/g for Ni(2+) at 298 K when the initial concentration for both Cu(2+) and Ni(2+) was 100mg/L, and the pH of the solution was 4.0 and 4.5, respectively. Column studies were conducted to generate breakthrough data for both pure component and binary mixtures of copper and nickel. Desorption experiments showed that 2mM EDTA solution could be used to remove all of the adsorbed copper and nickel from the bed. PMID- 18430508 TI - Agenesis of the internal carotid artery with transcavernous anastomosis presenting with an anterior communicating artery aneurysm--a case report and review of the literature. AB - Arterial communication between cavernous segments of the carotid arteries associated with unilateral agenesis of the internal carotid artery (ICA) is extremely rare. We herein present a case of unilateral ICA agenesis with transcavernous anastomosis that presents with an anterior communicating artery (ACoA) aneurysm, and discuss its embryogeny and clinical implications. A 55-year old woman with no significant medical history was admitted to our hospital for further examination of agenesis of the left ICA, which was detected in a routine medical examination. She was free of clinical symptoms and had no neurological deficits. Radiological evaluation revealed agenesis of the left ICA with transcavernous anastomosis. Moreover, an incidental multilobulated aneurysm was detected at the ACoA. She underwent microsurgical clipping for this aneurysm, and the post-operative course was uneventful. Investigation of collateral pathways helps our understanding of the segmental nature of carotid artery development. Recognition of this anomaly also has important implications in the surveillance and detection of associated cerebral aneurysms. PMID- 18430509 TI - Protoapigenone, a novel flavonoid, inhibits ovarian cancer cell growth in vitro and in vivo. AB - Flavonoids are polyphenolic compounds and capable of inhibiting the growth of human cancer cells. Protoapigenone, a novel flavonoid, was isolated from the whole plant Thelypteris torresiana (Gaud), a native fern in Taiwan. In the present study, we explored the cytotoxic effects of protoapigenone on ovarian cancer cells and the immortalized ovarian epithelial cells by XTT assay. The effects of protoapigenone on cell cycle progression and apoptosis were also analyzed by FACS analysis, immunofluorescence study and immunoblotting analysis. The anti-ovarian cancer effect of protoapigenone was further examined using nude mice xenograft assay and immunohistochemistry. Our results showed that protoapigenone had a significant cytotoxicity on human ovarian cancer cells MDAH 2774 and SKOV3 but not on the immortalized non-cancer ovarian epithelial cells HOSE 6-3 and HOSE 11-12. Protoapigenone arrested MDAH-2774 and SKOV3 cells at S and G2/M phases via decreasing the expression of p-Cdk2, Cdk2, p-Cyclin B1 and Cyclin B1, as well as increasing the expression of inactive p-Cdc25C. Besides, protoapigenone had an enhanced cytotoxicity on SKOV3 cells enriched at S and G2/M phases, and ability to induce apoptosis through decreasing the protein levels of Bcl-xL and Bcl-2 and increasing the cleaved PARP by activating caspase-3. In nude mice study, protoapigenone treatment significantly suppressed the tumor growth, without major side effects. Taken together, protoapigenone showed a significant anti-ovarian cancer activity with low toxicity, suggesting its potential to be developed as a chemotherapeutic agent. PMID- 18430510 TI - Magnetic removal of dyes from aqueous solution using multi-walled carbon nanotubes filled with Fe2O3 particles. AB - The Fe2O3 nanoparticles have been introduced into the multi-walled carbon nanotubes (MWCNTs) via wet chemical method. The resulting products are characterized by TEM, EDX, XRD and VSM. The magnetic MWCNTs have been employed as adsorbent for the magnetic separation of dye contaminants from water. The adsorption test of dyes (Methylene Blue and Neutral Red) demonstrates that it only takes 60min to attain equilibrium and the adsorption capacities for Methylene Blue and Neutral Red in the concentration range studied are 42.3 and 77.5mg/g, respectively. The magnetic MWCNTs can be easily manipulated in magnetic field for desired separation, leading to the removal of dyes from polluted water. The integration of MWCNTs with Fe2O3 nanoparticles has great potential application to remove organic dyes from polluted water. PMID- 18430511 TI - Occurrence of anionic surfactants in treated sewage: risk assessment to aquatic environment. AB - A comparative evaluation of occurrence of and risk to aquatic environment due to anionic surfactants (AS) in treated effluents from three main treatment processes, i.e. activated sludge process (ASP), oxidation pond (OP), and upflow anaerobic sludge blanket reactor (UASBR) is presented. UASBR effluents contained substantial concentrations of AS (4.25-5.91mg/L as average AS removal was not found to exceed 18%). Post-treatment of UASBR effluent using 1-1.6 days detention, anaerobic polishing ponds (PP) was also found quite ineffective. In UASBR-PP combine, AS reduced only up to 30%. Effluents from OP based sewage treatment plants (STPs) also contained significant concentrations of AS. On the contrary, effluent AS or linear alkylbenzene sulfonate (LAS) concentrations recorded in ASP effluents were quite low (less than 0.2mg/L). Unlike UASBR, LAS or AS removals greater than 99% are achieved in ASP. Treated effluents from UASBR and OP based STPs when discharged to aquatic ecosystems are likely to cause substantial risk to aquatic environment due to the presence of AS while effluents from ASP are not supposed to pose risk. Need to find an effective aerobic post treatment unit to UASBR for desired removal of AS is emphasized. PMID- 18430512 TI - Arsenic uptake by aquatic macrophyte Spirodela polyrhiza L.: interactions with phosphate and iron. AB - The uptake of arsenate (As(V)) and dimethylarsinic acid (DMAA) by aquatic macrophyte Spirodela polyrhiza L. was investigated to determine the influence of arsenic interaction with PO4(3-) and Fe ions. Plants were grown hydroponically on standard Murashige and Skoog (MS) culture solutions. Arsenic concentrations in Fe oxide (Fe-plaque) on plant surfaces were determined by citrate-bicarbonate ethylenediaminetetraacetic acid (CBE) technique. S. polyrhiza L. accumulated 51 fold arsenic from arsenate solution compared to that from DMAA solution with initial concentrations of 4.0 and 0.02microM of arsenic and phosphate, respectively. The arsenate uptake was negatively (p<0.001) correlated with phosphate uptake and positively (p<0.05) correlated with iron uptake. About 56% of the total arsenic was accumulated into the plant tissues while 44% was adsorbed on Fe-plaque (CBE-extract), when the plants were grown on arsenate solution. The DMAA uptake into the plant was neither affected by the phosphate concentrations nor correlated (p>0.05) with iron accumulation. The results suggest that adsorption of arsenate on Fe-plaque of the surface of S. polyrhiza L. contributes to the arsenic uptake significantly. Thus, arsenate uptake in S. polyrhiza L. occurred through the phosphate uptake pathway and by physico chemical adsorption on Fe-plaques of plant surfaces as well. The S. polyrhiza L. uses different mechanisms for DMAA uptake. PMID- 18430513 TI - Studies on efficiency of guava (Psidium guajava) bark as bioadsorbent for removal of Hg (II) from aqueous solutions. AB - Biosorption of Hg (II) was investigated in this study by using guava bark powder (GBP). In the batch system, effects of various parameters like contact time, initial concentration, pH and temperature were investigated. Removal of Hg (II) was pH dependent and was found maximum at pH 9.0. Based on this study, the thermodynamic parameters like change in standard Gibb's free energy (DeltaG(0)), standard enthalpy (DeltaH(0)) and standard entropy (DeltaS(0)) were evaluated. The rate kinetic study was found to follow second-order. The applicability of Freundlich adsorption isotherm model was tested. The value of regression coefficient was greater than 0.99. This indicated that the isotherm model adequately described the experimental data of the biosorption of Hg (II). Maximum adsorption of 3.364 mgg(-1) was reached at 80 min. The results of the study showed that guava bark powder can be efficiently used as a low-cost alternative for the removal of divalent mercury from aqueous solutions. PMID- 18430514 TI - Preparation and lead ion removal property of hydroxyapatite/polyacrylamide composite hydrogels. AB - We report the synthesis of hydroxyapatite/polyacrylamide (HAp/PAAm) composite hydrogels with various HAp contents by free radical polymerization and their removal capability of Pb(2+) ions in aqueous solutions with controlled initial Pb(2+) ion concentrations and pH values of 2-5. The swelling ratio of the composite gels in aqueous solutions decreases with increasing the HAp content in the gels. The composite gel with higher HAp content exhibits the higher removal capacity of Pb(2+) ions owing to the higher adsorption sites for Pb(2+) ions, but shows the slower removal rate of Pb(2+) ions due to the lower degree of swelling. The removal mechanism of Pb(2+) ion is very sensitive to the pH value in aqueous solution, although the removed amount of Pb(2+) ion is nearly same, regardless of pH values of 2-5. The removal mechanism, the dissolution of HAp in the composite gel and subsequent precipitation of hydroxypyromorphite (HPy), is dominant at lower pH 2-3, whereas the mechanism, the adsorption of Pb(2+) ions on the composite gel and following cation exchange reaction between Pb(2+) ions adsorbed and Ca(2+) of HAp, is dominant at higher pH 4-5. The equilibrium removal process of Pb(2+) ions by the composite gels at pH 5 is described well with the Langmuir isotherm model. The equilibrium removal capacities of the composite gels with 30, 50, and 70 wt.% HAp contents are evaluated to be 123, 178, and 209 mg/g, respectively. PMID- 18430515 TI - Bioleaching of spent Ni-Cd batteries by continuous flow system: effect of hydraulic retention time and process load. AB - Spent Ni-Cd batteries bring a severe environmental problem that needs to be solved urgently. A novel continuous flow two-step leaching system based on bioleaching was introduced to dissolve heavy metals in batteries. It consists of an acidifying reactor which was used to culture indigenous thiobacilli and a leaching reactor which was used to leach metals from spent batteries. The indigenous acidophilic thiobacilli in sewage sludge was used as the microorganisms and the sludge itself as culture medium. Bioleaching tests at different hydraulic retention time (HRT) and process load in the leaching reactor were performed. The results showed that the longer the HRT (1, 3, 6, 9 and 15 days) was, the more time required to achieve the complete leaching of Ni, Cd and Co. The maximum dissolution of cadmium and cobalt was achieved at higher pH values (3.0-4.5) while the leaching of nickel hydroxide and nickel in metallic form (Ni0) were obtained separately in different acidity (pH 2.5-3.5). It cost about 25, 30 and more than 40 days to remove all of the three heavy metals with the process load of two, four and eight Ni-Cd batteries under the conditions that the ingoing bio-sulphuric acid was 1Ld(-1) and HRT was 3 days. PMID- 18430516 TI - Alternatively spliced human catechol-O-methyltransferase (COMT) rationalizes lack of Val158Met effects on pain tolerance. PMID- 18430517 TI - Stretching it for pain. PMID- 18430518 TI - Psychological predictors of pain expression and activity intolerance in chronic pain patients. AB - Recent research suggests that communicative and protective pain behaviors represent functionally distinct subsystems of behavior associated with pain. The present research examined whether components of pain experience such as pain severity, catastrophizing and fear of pain were differentially associated with communicative and protective pain behaviors. It was predicted that pain severity would be associated with decreased physical tolerance and heightened expression of pain behavior. It was also predicted that pain catastrophizing would be preferentially associated with communicative pain behaviors, and fear of pain would be preferentially associated with protective pain behaviors and decreased physical tolerance. To test these predictions, work-disabled patients with musculoskeletal pain conditions (N=72) were filmed as they participated in a simulated occupational lifting task. Multiple regressions revealed that pain severity was uniquely associated with decreased physical tolerance and increased expression of protective pain behaviors. Pain catastrophizing was uniquely associated with the expression of both communicative and protective pain behaviors. Fear of pain was associated with physical tolerance and protective pain behaviors but not when controlling for pain severity. This study provides additional evidence for the functional distinctiveness of different types of pain expression and provides preliminary evidence for the functional distinctiveness of pain expression and activity intolerance. Discussion addresses the processes by which psychological factors might influence the display of different types of pain behaviors. Discussion also addresses how different types of interventions might be required to specifically target the sensory and behavioral dimensions of the pain system. PMID- 18430519 TI - Sarcoptic mange in red deer from Spain: improved surveillance or disease emergence? AB - Concern about emerging diseases has risen in recent years, and multihost situations have become increasingly relevant for wildlife management and conservation. We present data on Asturias, northern Spain, where 80 mangy red deer (Cervus elaphus) have been found since the beginning of the epizootic in chamois (Rupicapra pyrenaica parva) in 1993. We combine field and necropsy data with the results of a serosurvey using an in-house ELISA test to evaluate if deer mange due to Sarcoptes scabiei is an emerging disease in this area. The mean number of deer mange cases per year was 5, with a maximum of 16. No significant relationship was detected between monthly temperatures, rainfall or number of days with snow cover and the annual number of sarcoptic mange cases in red deer. Only 4 mangy red deer (5%) were detected outside the limits of scabietic chamois distribution during the same year, and all were less than 2500 m away from that limit. The longest distance reported between two consecutive mangy deer locations was 18 km. Mange cases were significantly more frequent in stags than in hinds and in adults than in juvenile deer. The time of the first mange detection in chamois in each sector, year with minimum number of chamois recorded, year with maximum chamois population decline rate and chamois density offered no significant correlation with red deer mange cases appearance moment and frequency. In the mange affected area, ELISA testing of 327 blood samples from hunter-harvested deer without obvious mange-compatible lesions revealed only 4 seropositive animals. All 83 sera from hunting preserves without clinical cases yielded negative ELISA results. According to these epidemiological data mange does not seem to threaten red deer populations in Asturias. However, continued monitoring of deer health and ELISA testing for sarcoptic mange is advisable. PMID- 18430520 TI - Prevalence of recent illicit substance use and reporting bias among MSM and other urban males. AB - This paper explores whether elevated rates of self-reported substance use among MSM compared to other males may be an artifact of reporting bias. Past month prevalence rates of marijuana, cocaine, heroin, methamphetamine, Ecstasy, and Ketamine use were compared between a sample of men who have sex with men (MSM), and a general household sample of men, all residing in Chicago. We compared rates of self-reported use, and corrected rates based on the results of drug testing (urine and oral fluid tests). While MSM over 30 years old were significantly more likely than other men in this age group to report past month use of cocaine, test corrected rates of use were equivalent. On the other hand, test-corrected estimates confirmed elevated rates of Ketamine and Ecstasy use in the MSM sample. Differential disclosure of substance use between MSM and other males may in some cases lead to distorted conclusions about differences in substance use between these groups. The use of biological testing in epidemiological studies of substance use can reduce the uncertainty of such comparisons. PMID- 18430521 TI - Accuracy of the local effect model for the prediction of biologic effects of carbon ion beams in vitro and in vivo. AB - PURPOSE: To analyze the accuracy of relative biologic effectiveness (RBE) values for treatment planning in carbon ion radiotherapy based on the local effect model (LEM) and to discuss the implications on the clinically relevant depth dose profiles. METHODS AND MATERIALS: Predictions of the LEM are compared with a broad panel of experimental data in vitro and to the tolerance of the rat spinal cord in vivo. To improve the accuracy of the LEM, the description of track structure is modified by taking into account a velocity-dependent extension of the inner part of the track. RESULTS: The original version of the LEM (LEM I) underestimates the therapeutic ratio of carbon ions (i.e., the ratio of RBE in the Bragg peak region as compared with the RBE in the entrance channel). Although significantly reduced, the cluster extension of the LEM (LEM II) still shows the same tendency. Implementation of the modified track structure (LEM III) almost completely compensates these systematic deviations, and predictions of RBE by LEM III for high and low energetic carbon ions show good agreement for a wide panel of different cell lines, as well as for the tolerance of the rat spinal cord. As a consequence, the expected RBE in the normal tissue surrounding the tumor becomes significantly lower than estimated with the LEM in its original version (LEM I). CONCLUSIONS: The modified track structure description represents an empiric approach to improve the accuracy of the LEM for treatment planning. This will be particularly useful for further optimization of carbon ion therapy in general and with respect to comparison with other treatment modalities, such as protons or intensity-modulated radiotherapy. PMID- 18430522 TI - Social reward-conditioned place preference: a model revealing an interaction between cocaine and social context rewards in rats. AB - A recent thrust in drug abuse research is the influence of social interactions on drug effects. Therefore, the present study examined conditioned place preference (CPP) as a model for assessing interactions between drug and social rewards in adolescent rats. Parameters for establishing social reward-CPP were examined, including the number of conditioning sessions/day (1 or 2), the total number of sessions (2, 8, or 16), and the duration of sessions (10 or 30 min). Subsequently, the effects of cocaine or dextromethorphan on social reward-CPP and play behavior were examined. The results demonstrate that social reward-CPP (i.e., preference shift for an environment paired previously with a rat) was similar using either 1 or 2 conditioning sessions/day and either 10 or 30 min sessions; however, social reward-CPP increased as the number of social pairings increased. Additionally, a low dose of cocaine (2 mg/kg, IP) and a low number of social pairings (2 pairings) failed to produce CPP when examined alone, but together produced a robust CPP, demonstrating an interaction between these rewards. The non-rewarding drug, dextromethorphan (30 mg/kg, IP), failed to enhance social reward-CPP, suggesting that drug-enhanced social reward-CPP is specific to rewarding drugs. Surprisingly, there was no relationship between play behaviors and preference shift in drug-naive animals. Furthermore, cocaine inhibited play behavior despite enhancing social reward-CPP, suggesting that aspects of social interaction other than play behavior likely contribute to social reward. The findings have important implications for understanding the influence of social context on cocaine reward during adolescence. PMID- 18430523 TI - Drinkers and bettors: investigating the complementarity of alcohol consumption and problem gambling. AB - Regulated gambling is a multi-billion dollar industry in the United States with greater than 100% increases in revenue over the past decade. Along with this rise in gambling popularity and gaming options comes an increased risk of addiction and the associated social costs. This paper focuses on the effect of alcohol use on gambling-related problems. Variables correlated with both alcohol use and gambling may be difficult to observe, and the inability to include these items in empirical models may bias coefficient estimates. After addressing the endogeneity of alcohol use when appropriate, we find strong evidence that problematic gambling and alcohol consumption are complementary activities. PMID- 18430524 TI - Diagnostic orphans for alcohol use disorders in a treatment-seeking psychiatric sample. AB - Individuals who endorse one or two of the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition (DSM-IV) criterion items for alcohol dependence but do not meet criteria for either alcohol abuse or dependence have been referred to in the literature as "diagnostic orphans." The goal of the present study is to compare diagnostic orphans for alcohol use disorders (AUD) to patients with lifetime DSM-IV alcohol abuse, alcohol dependence, and those with no-AUD symptoms on a series of demographic, diagnostic, and clinical measures. Participants were treatment-seeking psychiatric outpatients (n=1793; 61.5% women) who completed an in-depth, face-to-face diagnostic evaluation for DSM-IV axis I and axis II disorders. Results revealed that diagnostic orphans were younger, had a higher frequency of family history positive for alcoholism, and higher rates of cannabis dependence, as compared to the no-AUD symptoms group. Diagnostic orphans differed significantly from patients with alcohol abuse and dependence on a number of demographic, diagnostic, and clinical measures. Most notably, on a lifetime basis, diagnostic orphans were less likely to meet diagnostic criteria for various substance use disorders, as compared to individuals with alcohol abuse and dependence. Taken together, these results generally do not support combining diagnostic orphans to individuals with alcohol abuse. PMID- 18430525 TI - Effect of porcine circovirus type 2 (PCV2) vaccination on porcine reproductive and respiratory syndrome virus (PRRSV) and PCV2 coinfection. AB - The objectives were to determine if PCV2 vaccination is effective in reducing disease and lesions associated with PRRSV and PCV2 coinfection and if there is a difference between intradermal (ID) and intramuscular (IM) route of PCV2 vaccination. Seventy-four, 21-day-old pigs were randomly allocated into one of six groups. On day 0, pigs were vaccinated with 2ml Suvaxyn PCV2 One Dose (Fort Dodge Animal Health, Inc.) by intramuscular (VAC-M-COINF) or intradermal (VAC-D COINF) routes. On day 28, pigs were either singularly (PRRSV-only, PCV2-only) or coinfected (COINF) with PRRSV and PCV2. All pigs in all groups were necropsied on day 42. All vaccinated pigs seroconverted (IgM, IgG, and neutralizing antibodies) to PCV2 between 14 and 28 days post-vaccination. After challenge, all groups inoculated with PRRSV had reduced average daily gain compared to CONTROLS and PCV2-only (P<0.001). COINF pigs had significantly (P<0.05) reduced anti-PCV2-IgG antibody levels and neutralizing antibody levels compared to both vaccinated groups. COINF pigs had more severe lung lesions compared to VAC-M-COINF (P<0.05). COINF pigs had higher amounts of PCV2 DNA in serum samples and feces (P<0.05) and increased amounts of PCV2 in lymphoid tissues (P<0.05) compared to both vaccinated groups. In summary, PCV2 vaccination was effective at inducing a neutralizing antibody response and significantly reducing PCV2-associated lesions and PCV2 viremia in pigs coinfected with PCV2 and PRRSV. Differences between intradermal and intramuscular routes of vaccine administration were not observed. PMID- 18430526 TI - Testing the efficacy of medium chain fatty acids against rabbit colibacillosis. AB - Enteropathogenic Escherichia coli (EPEC) represents a major cause of lethal diarrhea in young mammals. Although the pathogenicity mechanisms of EPEC are now well understood, the intrinsic and environmental factors that control the expression of EPEC virulence remain largely unknown. In the rabbit, suckling reduces pups' sensitivity to EPEC infection. Hence, we have hypothesized that uncharacterized factors present in doe milk may mediate this protection. Medium chain fatty acids (MCFA), known to possess antimicrobial properties, are highly abundant in doe milk. We demonstrate that caprylic acid exhibits a clear bacteriostatic effect in vitro against the rabbit EPEC strain E22 (O103:H2:K-), in a dose-dependent manner. In vivo, the dietary inclusion of triglycerides of MCFA did not however reduce the sensitivity of young rabbits challenged with this EPEC strain. The mortality and fecal excretion of EPEC were not reduced, and the bacterial adhesion to ileum was not inhibited. Amount of MCFA reaching the ileal level might have been too low and/or their association to other milk antimicrobials may have been required to observe a positive effect on disease evolution in a context of a highly virulent challenge. PMID- 18430527 TI - Glutathione S-transferase A1 polymorphism as a risk factor for smoking-related type 2 diabetes among Japanese. AB - Glutathione S-transferases protect cells against exogenous and endogenous oxidative stress. Type 2 diabetes is associated with an increased production of reactive oxygen species and a reduction in antioxidant defenses. This study investigated whether GSTA1*A/*B and GSTP1Ile105Val polymorphisms could affect the risk for type 2 diabetes. A cross-sectional case-control analysis included 468 (326 men and 142 women) Japanese participants in a health screening program. The prevalence of type 2 diabetes was 11.3% (63 subjects: 52 male and 11 female). The frequency of GSTA1*B allele carriers was higher in diabetes than in non-diabetes, though the difference was not statistically significant (adjusted OR, 1.8; 95% CI, 0.9-3.4). The risk among the GSTA1*B allele carriers was significantly increased by current-smoking status (adjusted OR, 3.7; 95% CI, 1.1-12.7; vs. never-smoking non-carriers), whereas the smoking status was not an independent risk factor. The GSTP1 genotype alone or in combination with the smoking status did not affect the risk for diabetes. This is the first report to show that the GSTA1*B allele is a potential risk factor for smoking-related type 2 diabetes. PMID- 18430528 TI - Inhalation and epidermal exposure of volunteers to ethylene glycol: kinetics of absorption, urinary excretion, and metabolism to glycolate and oxalate. AB - Ethylene glycol (EG) is a widely used liquid. Limited data are published regarding inhaled EG and no data regarding transdermal EG uptake in humans. In order to gain information on the quantitative fate of EG, four male volunteers inhaled between 1340 and 1610 micromol vaporous 13C-labeled EG (13C2-EG) for 4h. Separately, three of these subjects were epidermally exposed for up to 6h to liquid 13C2-EG (skin area 66 cm2). Plasma concentrations and urinary amounts of 13C2-EG were determined by gas chromatography with mass selective detection. Additionally, plasma was assayed for 13C-labeled glycolic acid 13C2-GA) and urine for 13C2-GA and 13C-labeled oxalic acid (13C2-OA). Both EG metabolites were nephrotoxic in animals and humans and embryotoxic in rodents. 13C-labels enabled to differentiate from also determined endogenous EG, glycolic acid (GA), and oxalic acid (OA). Of 13C2-EG inhaled, 5.5+/-3.0%, 0.77+/-0.15%, and 0.10+/-0.12% were detected in urine as 13C2-EG, 13C2-GA, and 13C2-OA, respectively. The skin permeability constant of liquid EG was 2.7 x 10(-5)+/-0.5 x 10(-5)cm/h. Of the dose taken up transdermally, 8.1+/-3.2% and up to 0.4% were excreted in urine as 13C2-EG and 13C2-GA, respectively. It is calculated that equally long-lasting exposure to 10 ppm vaporous EG or wetting of both hands by liquid EG leads to about the same body burden by EG and metabolites. The amounts of GA and OA excreted daily in urine as a result of exposure (8h/day) to 10 ppm EG are about 15% and 2%, respectively, of those excreted from naturally occurring endogenous GA and OA. PMID- 18430529 TI - Formulation of monolayered films with drug and polymers of opposing solubilities. AB - The aim of this study was to prepare and characterise monolayered multipolymeric films (MMFs) comprising of a hydrophilic drug (Propranolol HCl) (PHCl) and polymers of opposing solubilities. Films were prepared by emulsification and casted by a new approach using a silicone-molded tray with individual wells. MMFs comprising of PHCl with Eudragit 100 (EUD100) and Chitosan (CHT), i.e. films with drug and polymers of opposing solubilities were successfully prepared (PHCl:EUD100:CHT; 1:10:0.5) and demonstrated uniform and reproducible drug content (100.71+/-2.66%), thickness (0.442+/-0.030 mm), mucoadhesivity (401.40+/ 30.73 mN) and a controlled drug release profile. Drug release followed Higuchi's square-root model. Maximum swelling of the films occurred after 1h and 28.26% of the films eroded during the 8-h test period. Mechanical testing revealed that the MMFs displayed a greater abrasion resistance, were more elastic and also required more energy to break, rendering them tougher and more suitable for buccal delivery than the monopolymeric PHCl:EUD100 film. The inclusion of CHT to the film led to a more porous surface morphology. The surface pH of the films remained constant at neutral pH. This study confirmed the potential of the above MMFs as a promising candidate for buccal delivery of PHCl. PMID- 18430530 TI - Response of the flat cochlear epithelium to forced expression of Atoh1. AB - Following hair cell elimination in severely traumatized cochleae, differentiated supporting cells are often replaced by a simple epithelium with cuboidal or flat appearance. Atoh1 (previously Math1) is a basic helix-loop-helix transcription factor critical to hair cell differentiation during mammalian embryogenesis. Forced expression of Atoh1 in the differentiated supporting cell population can induce transdifferentiation leading to hair cell regeneration. Here, we examined the outcome of adenovirus mediated over-expression of Atoh1 in the non-sensory cells of the flat epithelium. We determined that seven days after unilateral elimination of hair cells with neomycin, differentiated supporting cells are absent, replaced by a flat epithelium. Nerve processes were also missing from the auditory epithelium, with the exception of infrequent looping nerve processes above the habenula perforata. We then inoculated an adenovirus vector with Atoh1 insert into the scala media of the deafened cochlea. The inoculation resulted in upregulation of Atoh1 in the flat epithelium. However, two months after the inoculation, Atoh1-treated ears did not exhibit clear signs of hair cell regeneration. Combined with previous data on induction of supporting cell to hair cell transdifferentiation by forced expression of Atoh1, these results suggest that the presence of differentiated supporting cells in the organ of Corti is necessary for transdifferentiation to occur. PMID- 18430531 TI - Dissociation of procedural and semantic memory in absolute-pitch processing. AB - We describe two memory-retrieval systems in absolute-pitch (AP) processing and propose existence of a universal internal pitch template to which subpopulations of musicians selectively gain access through the two systems. In Experiment I, AP and control musicians adjusted the frequency of a pure tone to match the pitch of a visually displayed randomly selected musical note. In Experiment II the same subjects vocally produced within 2s the pitch associated with a randomly selected musical note label. AP musicians, but not controls, were highly accurate in frequency matching. Surprisingly, both AP and non-AP groups were extremely accurate in voicing the target pitch as determined from an FFT of the recorded voiced notes (i.e., sigma=0.97, 0.90 semitones, respectively). Spectrogram analysis showed that notes voiced by non-AP musicians are accurate from onset of voicing suggesting that pitch accuracy does not result from an auditory-motor feedback loop. Findings support existence of two memory-retrieval systems for musical pitch: a semantic associative form of memory used by AP musicians, and a more widespread form of procedural memory which allows precise access to internal pitch representations through the vocal-motor system. PMID- 18430532 TI - Gene expression analysis of human otosclerotic stapedial footplates. AB - Otosclerosis is a complex disease that results in a common form of conductive hearing loss due to impaired mobility of the stapes. Stapedial motion becomes compromised secondary to invasion of otosclerotic foci into the stapedio vestibular joint. Although environmental factors and genetic causes have been implicated in this process, the pathogenesis of otosclerosis remains poorly understood. To identify molecular contributors to otosclerosis we completed a microarray study of otosclerotic stapedial footplates. Stapes footplate samples from otosclerosis and control patients were used in the analysis. One-hundred-and ten genes were found to be differentially expressed in otosclerosis samples. Ontological analysis of differentially expressed genes in otosclerosis provides evidence for the involvement of a number of pathways in the disease process that include interleukin signaling, inflammation and signal transduction, suggesting that aberrant regulation of these pathways leads to abnormal bone remodeling. Functional analyses of genes from this study will enhance our understanding of the pathogenesis of this disease. PMID- 18430533 TI - Parents of childhood X-linked adrenoleukodystrophy: high risk for depression and neurosis. AB - The purpose of this study was to assess mental health in parents of patients with the childhood cerebral form of X-linked adrenoleukodystrophy (CCALD) and to investigate factors relating to psychological problems in order to improve clinical management and quality of life. Sixteen fathers and 21 mothers of patients with CCALD completed a battery of psychological examinations including the Beck Depression Inventory second edition (BDI-II), the General Health Questionnaire 60 (GHQ60), and the State-Trait Anxiety Inventory (STAI). Three fathers and 11 mothers showed high scores on the BDI-II, suggesting that they were in a depressive state. Depression in the mothers was serious as compared with previous reports. Six fathers and 11 mothers were considered to be in a state of neurosis, according to the results of the GHQ60. Four fathers and 8 mothers showed high levels of anxiety on the STAI. Health and social status of the mothers correlated with their mental health, and younger mothers with young patients tended to be more depressed. Thus, parents of patients with CCALD have a high risk of depression and neurosis. Understanding the mental state of these parents and improvements in the social support system including mental counseling, home nursing care, supports in workplace and community are necessary to prevent and treat psychological problems. Especially, early intervention for mental health problems should be provided for younger mothers with few years since the child's diagnosis. PMID- 18430534 TI - A 2-bp deletion in exon 74 of the dystrophin gene does not clearly induce muscle weakness. AB - Duchenne muscular dystrophy (DMD) is caused by mutation of the dystrophin gene. Cases of dystrophinopathy with a 2-bp deletion in the dystrophin gene commonly result in DMD. We report here a case of dystrophinopathy in a 9-years-old boy with a 2-bp deletion in exon 74 of the dystrophin gene; however, the boy had no clear clinical signs of muscle weakness. Immunohistochemical studies with N terminal (DYS3) and rod-domain anti-dystrophin (DYS1) antibodies revealed that the dystrophin signals were weaker than in the control sample (non dystrophinopathy) at the sarcolemma of myofibers, and the studies with C-terminus anti-dystrophin antibody (DYS2) were negative. Our patient's mutation is located between the binding sites of alpha-syntrophin and alpha-dystrobrevin. These results suggest that this mutation does not clearly induce muscle weakness at least through the age of 9 years. PMID- 18430537 TI - Shoulder ultrasound imaging-integrating anatomy, biomechanics and disease processes. AB - This article brings together the anatomy, biomechanics and the imaging of shoulder disease using ultrasound to enable a better understanding of the strengths and weaknesses of ultrasound when imaging the shoulder. PMID- 18430535 TI - Pituitary growth hormone suppression reduces resistin expression and enhances insulin effectiveness: relationship with caloric restriction. AB - Caloric restriction (CR) retards various age-dependent disorders, increases lifespan, and improves insulin activity in laboratory animals. Recently, adipocytes were found to act together as an active endocrine organ that produces various hormones called adipocytokines. The peripheral and central activities of these adipocytokines have been suggested to mediate the anti-aging effects of CR. Here, we tested this notion by analyzing the effect of CR and suppression of growth hormone/insulin-like growth factor-I (GH/IGF-I) axis on the expression of resistin, adiponectin, and adipsin genes by rat white adipose tissue (WAT). We found that CR and GH/IGF-I suppression markedly downregulated resistin gene expression. We also found plasma resistin levels correlated positively with pituitary GH mRNA expression levels. Our observations suggest that CR reduces resistin expression and increases insulin effectiveness in a GH/IGF-I-dependent manner. The beneficial effects of CR and GH/IGF-I suppression appear to be mediated, at least in part, by changes in glucose metabolism that result from reductions in plasma resistin levels. PMID- 18430538 TI - Diffusion-weighted whole-body MR screening. AB - Diffusion-weighted sequence (DWI) of the entire body is a new promising technique feasible to evaluate multifocal disease. DWI has revealed great potential in the evaluation of patients with cancer or benign disease, as it supplies both quantitative and qualitative information of the whole body. The technical aspects of the diffusion-weighted whole body (DWWB) MR sequence are described with special emphasis on the processing and analysis of the imaging. DWWB MR sequence should be used combined with the other standard sequences such as FSE T1-weighted and STIR images. A complete whole-body MR imaging protocol including the DWI can be performed in less than 40 min. The possibilities, limitations and the preliminary clinical results of the whole-body MR imaging using a DWI of the entire body are reviewed. PMID- 18430539 TI - Cryptosporidium--biotechnological advances in the detection, diagnosis and analysis of genetic variation. AB - Cryptosporidiosis is predominantly a gastrointestinal disease of humans and other animals, caused by various species of protozoan parasites representing the genus Cryptosporidium. This disease, transmitted mainly via the faecal-oral route (in water or food), is of major socioeconomic importance worldwide. The diagnosis and genetic characterization of the different species and population variants (usually recognised as "genotypes" or "subgenotypes") of Cryptosporidium is central to the prevention, surveillance and control of cryptosporidiosis, particularly given that there is presently no broadly applicable treatment regimen for this disease. Although traditional phenotypic techniques have had major limitations in the specific diagnosis of cryptosporidiosis, there have been major advances in the development of molecular analytical and diagnostic tools. This article provides a concise account of Cryptosporidium and cryptosporidiosis, and focuses mainly on recent advances in nucleic acid-based approaches for the diagnosis of cryptosporidiosis and analysis of genetic variation within and among species of Cryptosporidium. These advances represent a significant step toward an improved understanding of the epidemiology as well as the prevention and control of cryptosporidiosis. PMID- 18430540 TI - Venous complications following petrosal vein sectioning in surgery of petrous apex meningiomas. AB - OBJECTIVE: Meningiomas involving the petrous apex regularly show a close relationship with the superior petrosal vein which is sometimes obliterated during surgery due to its proximity to the tumour. However, there is no study available so far focusing on the frequency of postoperative venous congestion related complications following petrosal vein obliteration as well as on pre- and intraoperative findings related to them. METHODS: Fifty-nine patients with meningiomas involving the petrous apex were analyzed concerning the intraoperative preservation or sacrifice of the petrosal vein and postoperative complications related to venous occlusion. RESULTS: When a petrosal vein was occluded, in 9 of 30 cases venous-related complications occurred with a minor venous-congestion phenomenon in seven cases and major complications in two cases. When the petrosal vein complex was preserved, there were no similar complications. CONCLUSION: Preservation of the petrosal venous complex, especially of large caliber veins, should be attempted whenever possible to increase the safety of surgery. In cases of petrosal vein obliteration, effective brainstem decompression following tumour removal is essential to minimizing the risk of cerebellar congestion. PMID- 18430541 TI - [Paraspecific antivenins and exotic bites of snakes: about two case reports]. AB - Envenomation from bite of exotic snakes is rare in France and represent a serious therapeutic problem: only an adapted antivenom is effective and obtaining such a treatment is a real difficulty. The authors report two clinical cases of envenomation with defibrination after bite by pit vipers from South America. The two patients were treated with nonspecific antivenom therapy with divergent results. The use of the paraspecific effects of antivenom can allow an effective treatment in the absence of specific antivenom, but requires to be known better. PMID- 18430542 TI - [A first-degree heart block existed before anaesthesia and was only discovered after...]. PMID- 18430543 TI - WITHDRAWN: Treatment of combining antiangiogenic therapy and cytotoxic chemotherapy reduce the cancer stem-like cell fraction in hepatocarcinoma xenografts. AB - This article has been withdrawn consistent with Elsevier Policy on Article Withdrawal (http://www.elsevier.com/locate/withdrawalpolicy). The Publisher apologizes for any inconvenience this may cause. PMID- 18430544 TI - Roles of calcitonin gene-related peptide in facilitation of wound healing and angiogenesis. AB - Calcitonin gene-related peptide (CGRP) is a 37-amino acid neuropeptide produced by tissue-specific alternative splicing of the primary transcript of the calcitonin/CGRP gene. CGRP is widely distributed in the central and peripheral neuronal systems and exhibits numerous biological activities in mammals. We examined in the present study whether or not endogenous CGRP released from neuronal systems facilitates neovascularization indispensable to wound healing. In CGRP knockout mice (CGRP-/-), wound-induced angiogenesis and wound closure were significantly suppressed compared with those in wild-type mice. The suppressed healing in CGRP-/- was accompanied by reduction in expressions of vascular endothelial growth factor (VEGF) in the wound granulation tissues. A CGRP antagonist, CGRP8-37 when infused with mini-osmotic pumps subcutaneously blocked the wound healing processes and reduced the expressions of CD31 and VEGF expression in the wound granulation tissues. Wound healing process was significantly delayed in neuropeptide-depleted mice pretreated with capsaicin, compared with vehicle-treated mice. These results indicate that CGRP derived from neuronal systems may facilitate wound healing and angiogenesis. Targeting of CGRP may be promising in controlling angiogenesis related to pathophysiological conditions. PMID- 18430545 TI - Chronic obstructive pulmonary disease is not an independent marker for adverse outcomes in endograft repair of abdominal aortic aneurysms. AB - A significant proportion of patients undergoing endograft repair of abdominal aortic aneurysms (AAAs) suffer from chronic obstructive pulmonary disease (COPD). We report here our experience and analysis of 342 consecutive AAA endograft repairs in patients with and without COPD (137, or 39%, of patients with COPD and 55, or 16%, with moderate to severe COPD). Patient outcomes such as perioperative mortality, length of admission, intensive care unit admission, congestive heart failure, myocardial infarction, conversion to open surgery, duration of surgery, postoperative endoleaks, and combined respiratory complications were analyzed; differences were not statistically significant compared to patients without COPD (p > 0.05). Endograft repair of AAA demonstrated equivalent outcomes in patients with and without COPD. PMID- 18430547 TI - Enhanced automatic artifact detection based on independent component analysis and Renyi's entropy. AB - Artifacts are disturbances that may occur during signal acquisition and may affect their processing. The aim of this paper is to propose a technique for automatically detecting artifacts from the electroencephalographic (EEG) recordings. In particular, a technique based on both Independent Component Analysis (ICA) to extract artifactual signals and on Renyi's entropy to automatically detect them is presented. This technique is compared to the widely known approach based on ICA and the joint use of kurtosis and Shannon's entropy. The novel processing technique is shown to detect on average 92.6% of the artifactual signals against the average 68.7% of the previous technique on the studied available database. Moreover, Renyi's entropy is shown to be able to detect muscle and very low frequency activity as well as to discriminate them from other kinds of artifacts. In order to achieve an efficient rejection of the artifacts while minimizing the information loss, future efforts will be devoted to the improvement of blind artifact separation from EEG in order to ensure a very efficient isolation of the artifactual activity from any signals deriving from other brain tasks. PMID- 18430546 TI - Cortical dopaminergic neurotransmission in rats intoxicated with lead during pregnancy. Nitric oxide and hydroxyl radicals formation involvement. AB - It is well established that low level Pb-exposure is associated with a wide range of cognitive and neurobehavioral dysfunctions in children. In fact, Pb-induced damage occurs preferentially in the prefrontal cerebral cortex, hippocampus and cerebellum - the anatomical sites which are crucial in modulating emotional response, memory and learning. Previously it was also shown that nitric oxide (NO) signaling pathway as well as glutamatergic neurotransmission are both involved in brain development, neurotoxicity and neurodegeneration processes whereas Pb(2+) interfere with both. For this reason we investigated the effect of ontogenetic Pb(2+) exposure on dopaminergic neurotransmission in the medial prefrontal cortex (mPFC) of rats after amphetamine (AMPH) and/or 7-nitroindazole (7-NI) administration. Furthermore, the possible role of oxidative stress in Pb(2+)-induced neurotoxicity in prenatally Pb(2+)-treated rats was explored in the content of hydroxyl radical (HO) species in mPFC after AMPH and/or 7-NI injection, assessed by HPLC analysis of 2.3-dihydroxybenzoic acid (2.3-DHBA) - spin trap product of salicylate. As shown, the results of this study suggest that Pb(2+) exposure during intrauterine life did not substantially affect cortical dopaminergic neurotransmission in adult offspring rats evaluated by means of microdialysis of mPFC and the content of the cortical HO. It is likely that striatum, nucleus accumbens or other dopamine rich brain areas are more intricately associated with Pb(2+) precipitated behavioral, dopamine - dependent impairments observed in mammalians. PMID- 18430548 TI - Formation of shellac succinate having improved enteric film properties through dry media reaction. AB - The aim of this study was to improve enteric properties of shellac by the formation of succinate derivative through dry media reaction. Shellac and succinic anhydride were mixed and then co-ground by planetary ball mill. The ground mixture was then activated by heating for various times and washed for removal of excess succinic anhydride. The ground mixtures and the heat-activated mixtures were characterized by physical and chemical tests, including acid value, FTIR spectroscopy, (1)H NMR and (13)C NMR spectroscopy, thermal analysis and film properties. The results demonstrated that acid values of heat-activated shellac mixtures increased with the increase of annealing time, suggesting the presence of carboxylic acid moieties of succinate at shellac molecules. The results were in good agreement with the DSC thermograms. The melting peak of shellac disappeared after heating, while melting peak of succinic anhydride gradually decreased, suggesting the utilization of succinic anhydride for the esterification. The shellac succinate formation was also confirmed by (1)H NMR and (13)C NMR spectroscopies. Film prepared from shellac succinate showed improved solubility, especially at the pH of small intestine (5.8-6.7), as compared to native shellac. The shellac succinate film also demonstrated better mechanical property, in terms of increased flexibility. In conclusion, solid state formation of shellac succinate ester, which had improved enteric properties, was easily accomplished under the concept of "green approach". PMID- 18430549 TI - Formation mechanism and release behavior of poly(epsilon-caprolactone) microspheres containing disodium norcantharidate. AB - Disodium norcantharidate (DSNC) loaded poly(epsilon-caprolactone) (PCL) microspheres were prepared by s/o/w solvent evaporation technique, and the formation mechanism and release behavior of the microspheres were investigated. The particle formation of the microspheres was influenced by the osmotic effect of DSNC. During the microsphere preparation, water diffused into the emulsion droplets and dissolved the particles of DSNC. Thereafter, DSNC generated osmotic effect and drove the water to flow in the emulsion droplets more quickly, thus forming an inner water phase. As the water influx proceeded, the state of the emulsion was transferred from s/o/w to w/o/w, thus resulting in the porosity of the microspheres. The release tests were carried out in the release media of different osmotic pressures achieved by adding different amounts of dextrose. The results indicated that the initial release of DSNC from the microspheres was controlled by a combination of osmotic effect and diffusion, but the release after the initial was mainly controlled by diffusion. This study demonstrated that the osmotic effect of DSNC not only was responsible for the particle formation but also contributed to the release from the microspheres. PMID- 18430550 TI - Development of novel polymeric micellar drug conjugates and nano-containers with hydrolyzable core structure for doxorubicin delivery. AB - Novel micelle-forming poly(ethylene oxide)-block-poly(epsilon-caprolactone) (PEO b-PCL) block copolymers bearing doxorubicin (DOX) side groups (PEO-b-P(CL-DOX)) on the PCL block were synthesized. Prepared block copolymers were characterized, assembled to polymeric micellar drug conjugates and assessed for the level of DOX release at pH 7.4 and pH 5.0 using a dialysis membrane to separate released and conjugated drug. The possibility for the degradation of PCL backbone for PEO-b P(CL-DOX) micelles was investigated using gel permeation chromatography. Micelle forming DOX conjugate did not show any signs of DOX release at 37 degrees C within 72h of incubation at both pHs, but revealed signs of poly(ester) core degradation at pH 5.0. In further studies, PEO-b-PCL micelles bearing benzyl, carboxyl or DOX groups in the core were also used as micellar nano-containers for the physical encapsulation of DOX, where maximum level of drug-loading and control over the rate of DOX release was achieved by polymeric micelles containing benzyl groups in their core, i.e., PEO-b-poly(alpha-benzylcarboxylate epsilon-caprolactone) (PEO-b-PBCL) micelles. The in vitro cytotoxicity of chemically conjugated DOX as part of PEO-b-P(CL-DOX) and physically encapsulated DOX in PEO-b-PBCL against B16F10 murine melanoma cells was assessed and compared to that of free DOX. Consistent with the results of in vitro release study, cytotoxicity of micellar PEO-b-P(CL-DOX) conjugate (IC50 of 3.65 microg/mL) was lower than that of free and physically encapsulated DOX in PEO-b-PBCL (IC50 of 0.09 and 3.07 microg/mL, respectively) after 24 h of incubation. After 48 h of incubation, the cytotoxicity of conjugated DOX (IC50 of 0.50 microg/mL) was still lower than the cytotoxicity of free DOX (IC50 of 0.03 microg/mL), but surpassed that of physically encapsulated DOX in PEO-b-PBCL (IC50 of 1.54 microg/mL). The results point to a potential for PEO-b-P(CL-DOX) and PEO-b-PBCL as novel polymeric micellar drug conjugates and nano-containers bearing hydrolyzable cores for DOX delivery. PMID- 18430551 TI - Cytotoxicity of the beta-carboline alkaloids harmine and harmaline in human cell assays in vitro. AB - beta-Carboline alkaloids are natural products widely distributed in plants and also found in alcoholic beverages, well-cooked foods and tobacco smoke. Various authors have reported genotoxic activities of several carboline in prokaryotic and eukaryotic cells that have been attributed to their abilities to intercalate into DNA. But studies on the genotoxic and on the cytotoxic potencies in human cells in vitro are not found in the literature. In the present study the toxicities of one full aromatic beta-carboline alkaloid (harmine) and one dihydro beta-carboline alkaloid (harmaline) were evaluated by means of two in vitro human cell assays: the cytochalasin-B blocked micronucleus (CBMN) assay and the viability/colony formation assay with four different human cultured non transformed (CCD18Lu) and transformed (HeLa, C33A and SW480) cells. Neither alkaloid was able to induce micronuclei levels above that of control levels in a wide range of doses tested; although, harmine at the highest concentrations assayed induced apoptotic as well as necrotic cells. Harmine produced a good viability of all cell lines assayed (control and tumor) while harmaline significantly reduced the viability of transformed and non-transformed cell lines in a dose-dependent manner. Harmine displayed a dose-dependent inhibitory effect on cell proliferation against all human carcinoma cells, but the SW480 transformed cell line showed a higher sensitivity. These results suggested that harmine was identified as a useful inhibitor of tumor development. PMID- 18430552 TI - Regulative effects of essential oil from Atractylodes lancea on delayed gastric emptying in stress-induced rats. AB - Gastric motor dysfunction induced by psychological stress results in many symptoms of functional dyspepsia (FD). There are a number of herbal medicines that are reported to improve gastrointestinal motor. However, the mechanisms of considerable herbal medicines are not explicit. In the present study, the effects of an essential oil (EO) extracted from Atractylodes lancea on delayed gastric emptying, gastrointestinal hormone and hypothalamic corticotropin-releasing factor (CRF) abnormalities induced by restraint stress in rats were investigated and the mechanism of the EO is also explored. Oral administration of EO for 7 days did not affect normal gastric emptying, but accelerated delayed gastric emptying induced by restraint stress in rats. The EO significantly increased the levels of motilin (MTL) and gastrin (GAS) and decreased the levels of somatostatin (SS) and CRF. The EO did not modify gastric emptying in vagotomized rats that underwent restraint stress, but antagonized delayed gastric emptying induced by intracisternal injection of CRF. These results suggest that the regulative effects of the EO on delayed gastric emptying are preformed mainly via inhibition of the release of central CRF and activation of vagal pathway, which are also involved in the release of gastrointestinal hormones such as MTL, GAS and SS. PMID- 18430554 TI - Soybean diet improves insulin secretion through activation of cAMP/PKA pathway in rats. AB - Maternal malnutrition leads to permanent alterations in insulin secretion of offspring and the soybean diet contributes to improve insulin release. At least a soy component, genistein, seems to increase the insulin secretion by activating the cAMP/PKA and PLC/PKC pathways. Here, we investigated the effect of the soybean diet on the expression of PKAalpha and PKCalpha, and insulin secretion in response to glucose and activators of adenylate cyclase and PKC in adult pancreatic rat islets. Rats from mothers fed with 17% or 6% protein (casein) during pregnancy and lactation were maintained with 17% casein (CC and CR groups) or soybean (SC and SR groups) diet until 90 days of life. The soybean diet improved the insulin response to a physiological concentration of glucose in control islets, but only in the presence of supra-physiological concentrations of glucose in islets from CR and SR groups. PMA also improved the insulin response in islets of SC and SR groups. The expression of PKCalpha was similar in all groups. Forskolin increased the insulin secretion; however, the magnitude of the increment was lower in islets from CR and SR groups than in control animals and in those from rats maintained with soybean diet than in rats fed with casein diet. The PKAalpha expression was similar between SR and CR groups and lower in SC than in CC islets. Thus, soybean diet improved the secretory pattern of beta cells, at least in part, by activating the cAMP/PKA-signaling cascade. PMID- 18430553 TI - Sparc (Osteonectin) functions in morphogenesis of the pharyngeal skeleton and inner ear. AB - Sparc (Osteonectin), a matricellular glycoprotein expressed by many differentiated cells, is a major non-collagenous constituent of vertebrate bones. Recent studies indicate that Sparc expression appears early in development, although its function and regulation during embryogenesis are largely unknown. We cloned zebrafish sparc and investigated its role during development, using a mo rpholino antisense oligonucleotide-based knockdown approach. Consistent with its strong expression in the otic vesicle and developing pharyngeal cartilages, knockdown of Sparc function resulted in specific inner ear and cartilage defects that are highlighted by changes in gene expression, morphology and behavior. We rescued the knockdown phenotypes by co-injecting sparc mRNA, providing evidence that the knockdown phenotype is due specifically to impairment of Sparc function. A comparison of the phenotypes of Sparc knockdown and known zebrafish mutants with similar defects places Sparc downstream of sox9 in the genetic network that regulates development of the pharyngeal skeleton and inner ear of vertebrates. PMID- 18430556 TI - Synergistic induction of ER stress by homocysteine and beta-amyloid in SH-SY5Y cells. AB - Clinical studies have raised the possibility that elevated plasma levels of homocysteine increase the risk of atherosclerosis, stroke and possibly neurodegenerative diseases such as Alzheimer's disease (AD); however, the direct impact of homocysteine on neuron cells and the mechanism by which it could induce neurodegeneration have yet to be clearly demonstrated. Here, we investigated the effect of homocysteine on endoplasmic reticulum (ER) stress, the suggested mechanism of neurotoxicity, in human neuroblastoma SH-SY5Y cells. The effect of homocysteine on amyloid-beta (Abeta)-induced neurotoxicity and the protective activity of folate were also investigated. Homocysteine led to increased expressions of the binding protein (BiP) and the spliced form of X-box-protein (XBP)-1 mRNAs, suggesting activation of the unfolded-protein response and an increase in apoptosis. When cells were cotreated with homocysteine and Abeta, caspase-3 activity was significantly increased, and expressions of BiP and the spliced form of XBP-1 mRNAs were significantly induced. The neurotoxicity of homocysteine was attenuated by the treatment of cells with folate, as determined by caspase-3 activity and apoptotic body staining. These findings indicate that homocysteine induces ER stress and, ultimately, apoptosis and sensitizes neurons to amyloid toxicity via the synergistic induction of ER stress. Furthermore, a neuroprotective effect of folate against homocysteine-induced toxicity was also observed. Therefore, the findings of our study suggest that ER stress-induced homocysteine toxicity may play an important physiological role in enhancing the pathogenesis of Abeta-induced neuronal degeneration. PMID- 18430555 TI - Protein-energy malnutrition increases activation of the transcription factor, nuclear factor kappaB, in the gerbil hippocampus following global ischemia. AB - Protein-energy malnutrition (PEM) exacerbates functional impairment caused by brain ischemia. This is correlated with reactive gliosis, which suggests an increased inflammatory response. The objective of the current study was to investigate if PEM increases hippocampal activation of nuclear factor kappaB (NFkappaB), a transcription factor that amplifies the inflammatory response involved in ischemic brain injury. Mongolian gerbils (11-12 weeks old) were randomly assigned to control diet (12.5% protein) or protein-deficient diet (2%) for 4 weeks. The 2% protein group had a 15% decrease in voluntary food intake (P<.001; unpaired t test), resulting in PEM. Body weight after 4 weeks was 20% lower in the PEM group (P<.001). Gerbils were then exposed to sham surgery or global ischemia induced by 5-min bilateral common carotid artery occlusion. PEM independently increased hippocampal NFkappaB activation detected by electrophoretic mobility shift assay at 6 h after surgery (P=.014; 2-factor ANOVA). Ischemia did not significantly affect NFkappaB activation nor was there interaction between diet and ischemia. Serum glucose and cortisol concentrations at 6 h postischemia were unaltered by diet or ischemia. A second experiment using gerbils of the same age and feeding paradigm demonstrated that PEM also increases hippocampal NFkappaB activation in the absence of surgery. These findings suggest that PEM, which exists in 16% of elderly patients at admission for stroke, may worsen outcome by increasing activation of NFkappaB. Since PEM increased NFkappaB activation independent of ischemia or surgery, the data also have implications for the inflammatory response of the many individuals affected globally by PEM. PMID- 18430558 TI - Dietary conjugated linoleic acid in the cis-9, trans-11 isoform reduces parathyroid hormone in male, but not female, rats. AB - Previously, a mixture of conjugated linoleic acid (CLA) isoforms reduced parathyroid hormone (PTH) in male rats over 8 weeks. The objective herein was to determine which isoform caused the reduction in PTH; whether the effect was sex specific; and whether CLA-induced reductions in PTH were sustained. Male and female weanling rats (n=48) were randomized to a control diet or one made with 0.5% of the diet as cis-9, trans-11 (c9,t11) CLA, 0.5% of the diet as trans-10, cis-12 (t10,c12) CLA or these CLA in a mixture. Measurements made after 4, 8 and 16 weeks were body weight, bioactive PTH, ionized Ca, whole-body and regional bone mineral density (BMD) using dual-energy X-ray absorptiometry. With the use of a factorial design, a sexxc9,t11 CLA interaction was observed that reduced PTH (139.5+/-63.9 vs. 95.8+/-42.4 pg/ml, P=.02) in male rats only. No other effects of c9,t11 CLA were observed. Regarding t10,c12 CLA, no interaction effects were observed, but a main effect was observed to reduce lumbar spine BMD (0.265+/ 0.044 vs. 0.255+/-0.044 g/cm(2), P<.01) along with reduced retention of Ca and P at Week 4. No other dietary effects were observed. In summary, the c9,t11 CLA isoform is responsible for reduced PTH and this effect is sex specific; this was true whether fed as a pure isomer or mixed with an equal amount of t10,c12 CLA. Whether such reductions in PTH might be observed in females lacking sex hormones such as ovariectomized rats and also in humans is required to expand health implications of dietary CLA. PMID- 18430559 TI - Glucuronidation of the soyabean isoflavones genistein and daidzein by human liver is related to levels of UGT1A1 and UGT1A9 activity and alters isoflavone response in the MCF-7 human breast cancer cell line. AB - The soyabean isoflavones genistein and daidzein, which may protect against some cancers, cardiovascular disease and bone mineral loss, undergo substantial Phase 2 metabolism, predominantly glucuronidation. We observed a correlation between rates of metabolism of marker substrates of specific UGTs and rates of glucuronidation of genistein and daidzein in vitro by a panel of human liver microsomes, demonstrating that UGT1A1 and UGT1A9, but not UGT1A4, make a major contribution to the metabolism of these isoflavones by human liver. These findings were substantiated by observations that recombinant human UGT1A1 and UGT1A9, but not UGT1A4, catalysed the production of the major glucuronides of both genistein and daidzein in vitro. Recombinant human UGT1A8 also metabolised both genistein and daidzein, whereas UGT1A6 was specific to genistein and UGTs 2B7 and 2B15 were inactive, or only marginally active, with either isoflavone as substrate. The intestinal isoform UGT1A10 metabolised either both isoflavones or genistein only, depending on the commercial supplier of the recombinant enzyme, possibly as a result of a difference in amino acid sequence, which we were unable to confirm. Daidzein (16 microM) increased cell death in the MCF-7 human breast cancer cell line and this effect was reversed by glucuronidation. In view of a well-characterised functional polymorphism in UGT1A1, these observations may have implications for inter-individual variability in the potential health-beneficial effects of isoflavone consumption. PMID- 18430557 TI - The role of fatty acids in the development and progression of nonalcoholic fatty liver disease. AB - Nonalcoholic fatty liver disease (NAFLD) has emerged as a serious obesity-related disorder. NAFLD encompasses a wide spectrum of hepatic derangements ranging from a surfeit of fat in the liver (steatosis) to lipid surplus accompanied by fibrosis and cellular death (nonalcoholic steatohepatitis or NASH). The most widely accepted model to explain the progression from simple NAFLD to NASH is the "two-hit hypothesis," wherein fat over accumulation per se is not sufficient to induce the progression to statohepatitis, but renders the liver more susceptible to "second hits" that, once imposed upon the steatotic liver, cause further aberrations that culminate in the development of NASH. However, in light of recent data from our laboratory and elsewhere, we propose that an increased ratio of saturated-to-unsaturated fatty acids delivered to or stored within the liver may, in part, mediate the progression from simple steatosis to NASH. The molecular mechanisms that mediate the effect of saturated fatty acids are unclear, although proinflammatory cytokines, reactive oxygen species, and endoplasmic reticulum stress may all play a role. Collectively, these data suggest that saturated fatty acids may represent an intrinsic second hit to the liver that hastens the development of NASH. PMID- 18430560 TI - Aging and compressibility of municipal solid wastes. AB - The expansion of a municipal solid waste (MSW) landfill requires the ability to predict settlement behavior of the existing landfill. The practice of using a single compressibility value when performing a settlement analysis may lead to inaccurate predictions. This paper gives consideration to changes in the mechanical compressibility of MSW as a function of the fill age of MSW as well as the embedding depth of MSW. Borehole samples representative of various fill ages were obtained from five boreholes drilled to the bottom of the Qizhishan landfill in Suzhou, China. Thirty-one borehole samples were used to perform confined compression tests. Waste composition and volume-mass properties (i.e., unit weight, void ratio, and water content) were measured on all the samples. The test results showed that the compressible components of the MSW (i.e., organics, plastics, paper, wood and textiles) decreased with an increase in the fill age. The in situ void ratio of the MSW was shown to decrease with depth into the landfill. The compression index, Cc, was observed to decrease from 1.0 to 0.3 with depth into the landfill. Settlement analyses were performed on the existing landfill, demonstrating that the variation of MSW compressibility with fill age or depth should be taken into account in the settlement prediction. PMID- 18430561 TI - A novel biosensor based on activation effect of thiamine on the activity of pyruvate oxidase. AB - A biosensor based on pyruvate oxidase (POX) enzyme was developed for the investigation of the effect of thiamine (vitamin B(1)) molecule on the activity of the enzyme. The biosensor was prepared with a chemical covalent immobilization method on the dissolved oxygen (DO) probe by using gelatin and cross-linking agent, glutaraldehyde. POX catalyzes the degradation of pyruvate to acetylphosphate, CO(2) and H(2)O(2) in the presence of phosphate and oxygen. Thiamine is an activator for POX enzyme and determination method of the biosensor was based on this effect of thiamine on the activity of the enzyme. The biosensor responses showed increases in the presence of thiamine. Increases in the biosensor responses were related to thiamine concentration. Thiamine determination is based on the assay of the differences on the biosensor responses on the oxygenmeter in the absence and the presence of thiamine. The biosensor response depend linearly on thiamine concentration between 0.025 and 0.5 microM with 2 min response time. In the optimization studies of the biosensor the most suitable enzyme amount was found as 2.5 U cm(-2) and also phosphate buffer (pH 7.0; 50 mM) and 35 degrees C were obtained as the optimum working conditions. In the characterization studies of the biosensor some parameters such as activator and interference effects of some substances on the biosensor response and reproducibility were carried out. PMID- 18430562 TI - A chemiluminescent immunosensor based on antibody immobilized carboxylic resin beads coupled with micro-bubble accelerated immunoreaction for fast flow injection immunoassay. AB - A novel immunoaffinity column used as an immunosensor for flow-injection chemiluminescent (CL) immunoassay was prepared by immobilizing antibody on carboxylic resin beads. The immunosensor could fast recognize and trap the immunocomplex of horseradish peroxidase (HRP)-labeled antibody and antigen, which was firstly formed with a micro-bubble accelerated pre-incubation process, to produce a sandwich immunocomplex. The HRP introduced in the immunoaffinity column could catalyze the CL reaction to produce enzyme-enhanced emission. With alpha fetoprotein (AFP) as a mode, a flow-injection CL immunoassay was proposed. The whole assay for one sample, including the pre-incubation and the regeneration of immunoaffinity column, could be performed within 16 min. The linear range was 1.0 80 ng/ml with a correlation coefficient of 0.998 and a detection limit of 0.1 ng/ml at a signal/noise ratio of 3. The intra- and inter-assay coefficients of variation at 20 ng/ml AFP were 1.2% and 8.5%, respectively. The storage stability of the immunoaffinity column and the accuracy for sample detection were acceptable. This flexible, sensitive, low-cost, and rapid method is valuable for clinical immunoassay. PMID- 18430563 TI - Hospital differences in patient satisfaction with care for breast, colorectal, lung and prostate cancers. AB - BACKGROUND: We have investigated cancer patient satisfaction with care and the extent to which it varies between and within hospitals. DESIGN AND METHODS: A national survey of cancer patients in England with questions in 10 different dimensions for four common cancers: breast, colorectal, lung and prostate (55,674 patients). We compared hospitals across tumour types, and against the national average. RESULTS: Dissatisfaction was greater (p<0.001) in younger, female patients. Breast cancer patients expressed least, and prostate cancer patients expressed greatest dissatisfaction. Breast, colorectal and prostate cancers showed significant (p<0.001) pair-wise correlations for standardised satisfaction scores, particularly for in-hospital care. Summed hospital satisfaction scores showed significant associations across different dimensions of care. CONCLUSIONS: Cancer patient satisfaction is measurably different between hospitals, as well as by tumour type. For many aspects of care there is evidence of systemic hospital level factors that influence satisfaction as well as factors common to the care pathways experienced by individual patients. Factors amenable to clinical or managerial intervention deserve further investigation. PMID- 18430564 TI - Survival of children with cancer in Italy, 1989-98. A report from the hospital based registry of the Italian Association of Paediatric Haematology and Oncology (AIEOP). AB - We describe the survival patterns of 10,791 Italian children (age 0-14) diagnosed with cancer during 1989-1998 and who were included in the hospital-based registry of the Italian Association of Paediatric Haematology and Oncology. Five-year cumulative survival percentages were 76% for lymphoproliferative disorders and 68% for solid tumours. Survival rates in 1994-1998 significantly improved for acute lymphocytic leukaemia (ALL), acute non-lymphocytic leukaemia, Hodgkin's lymphoma and Wilms' tumour. Gender and age were determinants of survival for some specific types of cancer. Girls with ALL and neuroblastoma exhibited a significant advantage (hazard ratio HR 0.72, 0.62-0.83) and disadvantage (HR 0.73, 0.59-0.90) over boys, respectively. Children with a Wilms' tumour diagnosed above age 3 had a worse prognosis than younger children (HR 2.3, 1.4-4.1). The persisting gender-related difference in survival rate for ALL requires understanding as to whether it is attributable to delays in the adoption of more recent therapeutic protocols, while the corresponding findings for Wilms' tumour and neuroblastoma deserve further biological interpretation. PMID- 18430565 TI - Thyroid hormone and myocardial ischaemia. AB - Thyroid hormone has various effects on the cardiovascular system and its effects on cardiac contractility, heart rhythm and vascular function has long been recognized. However, new evidence is emerged on the importance of thyroid hormone in the response of the myocardium to ischaemic stress and cardiac remodelling following myocardial infarction. Based on this new information, this review highlights the role of thyroid hormone in myocardial ischaemia and cardiac remodelling, the possible underlying mechanisms and the potential therapeutic implications. Thyroid hormone or analogs may prove new therapeutic agents for treating ischaemic heart disease. PMID- 18430567 TI - Steroid binding sites in liver membranes: interplay between glucocorticoids, sex steroids, and pituitary hormones. AB - Steroid hormones activate target cells through specific receptors that discriminate among ligands based upon recognition of distinct structural features. For most known steroids, membrane and nuclear receptors co-exist in many target cells. However, while the structure of the nuclear receptors and their function as transcriptional activators of specific target genes is generally well understood, the identity of the membrane receptors remains elusive. Using pharmacological and biochemical approaches, we are beginning to characterize receptors for glucocorticoids and anabolic-androgenic steroids in male rat liver membranes. Male rat liver endoplasmic reticulum contains two steroid binding sites which are functionally related and associated with a 90-134 kDa oligomeric protein: (1) the low-affinity glucocorticoid binding site (LAGS), composed at least in part of two peptides (37 and 53 kDa) that bind glucocorticoids and (2) the stanozolol binding protein (STBP), composed at least in part of three peptides (22, 31, and 55 kDa) that bind the synthetic androgen stanozolol. These steroid binding proteins have many properties different from those of classical nuclear receptors, with the salient differences being a failure to recognize "classical" ligands for nuclear receptors together with marked differences in biochemical properties and physiological regulation. The mechanism of interaction of glucocorticoids with the LAGS can be clearly distinguished from that with STBP. Moreover, STBP shows an extremely narrow pharmacological profile, being selective for ST and its analog, danazol, among more than 100 steroids and non-steroidal compounds that were assayed, including those that are able to displace glucocorticoids from the LAGS. The level of LAGS activity undergoes dramatic variations following changes from the physiological serum levels of thyroid hormones, glucocorticoids, GH, vitamin A, and E2. However, neither thyroid hormones nor GH have a critical role on STBP activity. The STBP is functionally related to LAGS. We have suggested a novel mechanism for STBP whereby membrane-associated glucocorticoid binding activity is targeted by stanozolol (and 16beta-hydroxylated stanozolol): stanozolol modulates glucocorticoid activity in the liver through negative allosteric modulation of the LAGS resulting in an effective increase in classical GR-signaling by increasing glucocorticoid availability to the cytosolic GR. PMID- 18430566 TI - Cellular mechanisms of estradiol-mediated masculinization of the brain. AB - The sexual differentiation of reproductive physiology and behavior in the rodent brain is largely determined by estradiol aromatized from testicular androgens. The cellular mechanisms by which estradiol masculinizes the brain are beginning to emerge and revealing novel features of brain development that are highly region-specific. In the preoptic area, the major site controlling male sexual behavior, estradiol increases the level of the COX-2 enzyme and its product, prostaglandin E2 which promotes dendritic spine synaptogenesis. In the ventromedial nucleus of the hypothalamus, the major site controlling female reproductive behavior, estradiol promotes glutamate release from synaptic terminals, activating NMDA receptors and the MAP kinase pathway. In the arcuate nucleus, a major regulator of anterior pituitary function, estradiol increases GABA synthesis, altering the morphology of neighboring astrocytes and reducing formation of dendritic spines synapses. Glutamate, GABA and the importance of neuronal-astrocytic cross-talk are emerging as common aspects of masculinization. Advances are also being made in the mechanistic basis of female brain development, although the challenges are far greater. PMID- 18430568 TI - Optimization of media composition for Nattokinase production by Bacillus subtilis using response surface methodology. AB - Response surface methodology and central composite rotary design (CCRD) was employed to optimize a fermentation medium for the production of Nattokinase by Bacillus subtilis at pH 7.5. The four variables involved in this study were Glucose, Peptone, CaCl2, and MgSO4. The statistical analysis of the results showed that, in the range studied; only peptone had a significant effect on Nattokinase production. The optimized medium containing (%) Glucose: 1, Peptone: 5.5, MgSO4: 0.2 and CaCl2: 0.5 resulted in 2-fold increased level of Nattokinase (3194.25U/ml) production compared to initial level (1599.09U/ml) after 10h of fermentation. Nattokinase production was checked with fibrinolytic activity. PMID- 18430570 TI - Sulfamoyl benzamides as novel CB2 cannabinoid receptor ligands. AB - Sulfamoyl benzamides were identified as a novel series of cannabinoid receptor ligands. Starting from a screening hit 8 that had modest affinity for the cannabinoid CB(2) receptor, a parallel synthesis approach and initial SAR are described, leading to compound 27 with 120-fold functional selectivity for the CB(2) receptor. This compound produced robust antiallodynic activity in rodent models of postoperative pain and neuropathic pain without traditional cannabinergic side effects. PMID- 18430569 TI - Development of a fluorescence polarization based assay for histone deacetylase ligand discovery. AB - Histone deacetylases (HDACs) regulate many important physiological processes and the discovery of small molecules that modulate HDAC activity has both academic and clinical relevance. HDAC inhibitors, most notably SAHA, have been pursued as cancer chemotherapeutics but may be useful in treating psychiatric disorders, malaria, and other diseases. Herein, we describe an inexpensive and robust assay, based on fluorescence polarization, for HDAC ligand discovery. The assay is well suited for high-throughput screening and enzyme kinetic studies. PMID- 18430571 TI - Myopathy associated with chronic orlistat consumption: a case report. AB - The pathogenesis of myopathy due to HMG-CoA reductase inhibitors is not entirely clear but the existence of myopathy due to other lipid-lowering medications suggests it is mediated through effects on lipid metabolism. We describe a patient who developed a myopathy after 27 months of ingestion of orlistat that slowly improved after discontinuation of the drug. Orlistat may have caused this myopathy through depletion of a critical nutrient or through some as of yet undefined mechanism. PMID- 18430572 TI - No carnitine palmitoyltransferase deficiency in skeletal muscle in 18 malignant hyperthermia susceptible individuals. AB - Malignant hyperthermia is a rare, potentially life threatening pharmacogenetic disorder triggered by volatile anaesthetics and depolarizing muscle relaxants. The clinical picture comprises rhabdomyolysis, metabolic and respiratory acidosis, and hyperthermia. Carnitine palmitoyltransferase II deficiency is a metabolic myopathy affecting the transport of fatty acids into the mitochondria, leading to impaired energy supply under stressful conditions resulting in muscle weakness and rhabdomyolysis. It was postulated in a previous study that some patients with the MH phenotype have a carnitine palmitoyltransferase deficiency. To investigate a potential association, we tested 18 individuals with proven MH susceptibility for impairment of carnitine palmitoyltransferase enzyme activity in muscle. Enzyme activity was normal in all individuals tested indicating no impairment of the CPT system in this sample of malignant hyperthermia susceptible individuals. Thus our data do not support the hypothesis that susceptibility to malignant hyperthermia has an effect on the carnitine palmitoyltransferase enzyme system. PMID- 18430573 TI - Resistance to infection and the muscular dystrophies--is there a molecular link? PMID- 18430574 TI - Efficacy of Hylan G-F 20 and Sodium Hyaluronate in the treatment of osteoarthritis of the knee -- a prospective randomized clinical trial. AB - In this independent prospective randomized trial, we compared the clinical effectiveness, functional outcome and patient satisfaction following intra articular injection with two viscosupplementation agents - Hylan G-F-20 (n=199) and Sodium Hyaluronate (n=193) in patients with osteoarthritis (OA) of the knee. All patients were prospectively reviewed by blinded independent assessors at pre injection, 6 weeks, 3, 6, 12 months. Knee pain and patient satisfaction were measured on a visual analogue scale. Functional outcome was assessed using WOMAC, Oxford knee score and EuroQol EQ-5D scores. Knee pain on VAS improved from 6.7 to 3.2 by 6 weeks (p=0.02) and was sustained until 12 months (3.7, p=0.04) with Hylan G-F 20. In the Sodium Hyaluronate group, pain improved from 6.6 to 5.7 at 6 weeks (p>0.05) and to 4.1 at 3 months (p=0.04) but was sustained only until 6 months (5.9, p>0.05). Improvement in the WOMAC pain subscale was significantly superior in the Hylan G-F 20 group at 3 months (p=0.02), 6 months (p=0.01) and 12 months (p=0.007). There was no significant difference in the EQ-5D scores at 6 weeks and 3 months between the two groups. The numbers of treatment related adverse events were higher (39 vs. 30) in the Hylan G-F 20 group. One patient in the Hylan G-F 20 group who had a serious adverse event was also included in the final analysis. Although both treatments offered significant pain reduction, it was achieved earlier and sustained for a longer period with Hylan G-F 20. From this study, it appeared that the clinical effectiveness and general patient satisfaction are better amongst patients who received Hylan G-F 20. PMID- 18430575 TI - Petiolins A-C, phloroglucinol derivatives from Hypericum pseudopetiolatum var. kiusianum. AB - Two new phloroglucinol derivatives possessing chromane skeleton, petiolins A (1) and B (2), and a new phloroglucinol derivative containing a dihydrofuran ring, petiolin C (3), were isolated from aerial parts of Hypericum pseudopetiolatum var. kiusianum. The gross structures of 1-3 were elucidated by spectroscopic data, and the relative stereochemistry of 3 was elucidated by NOESY data. Petiolins A-C (1-3) showed modest cytotoxicity, while petiolin C (3) exhibited antifungal activity. PMID- 18430576 TI - Novel anti-inflammatory agents based on pyridazinone scaffold; design, synthesis and in vivo activity. AB - Herein, we report the design, synthesis, and pharmacological properties of a series of arylethenylpyridazinones 3a-h and arylethylpyridazinone derivatives 3k i from the corresponding aryloxohexenoic 1a-e and aryloxohexanoic acids 2a,d, respectively. The synthesized compounds were tested for their anti-inflammatory activity in carrageenan-induced rat paw edema model. Compound 3j demonstrated the greatest in vivo activity with ED(50) equal to 17mumol compared with celecoxib with no ulceration on the gastric mucosa. Docking study of the synthesized compounds into the active site of COX-2 revealed a similar binding mode to RS 57067, a COX-2 inhibitor. PMID- 18430577 TI - Micro-column solid phase extraction to determine uranium and thorium in environmental samples. AB - Extraction chromatographic separation techniques based on U/TEVA and TEVA resins were utilized to separate uranium and thorium isotopes in complex matrices from environmental samples. This approach has the advantages of ease of quantitative analysis, small sample size, an absence of mixed waste solvents, complete separation of U/Th isotopes, acceptable chemical yields and good energy resolution in the alpha spectrum. The procedure for analyzing alpha-emitting isotopes of uranium and thorium in geothermal water from Peito, Taiwan, is illustrated in detail. It involves sample pre-concentration, filtration and separation by highly selective extraction chromatographic resins, followed by electroplating and alpha-spectroscopy. The analytical results show a chemical recovery exceeding 55% for U and 65% for Th, respectively, under optimized conditions. The efficient and cost-effective use of recyclable columns makes the analytical methods simple, accurate, rapid, reliable and robust. PMID- 18430578 TI - Coincidence summing effects in gamma-ray spectrometry using a Marinelli beaker. AB - The coincidence summing effects on a HPGe spectrometer have been studied by using Marinelli beakers containing (133)Ba and (152)Eu solutions. The coincidence summing effects were calculated from the ratio of full-energy peak efficiency and total efficiency. The peak efficiency values for (133)Ba and (152)Eu corrected by using the peak-to-total ratio were found to be in good agreement (within 3%) as compared with the efficiency values from Marinelli beakers containing single gamma-ray emitters. PMID- 18430579 TI - A new method to quantify subtle morphological deformities in nasal profile curvatures and its application for analysis of unilateral cleft lip noses. AB - OBJECTIVE: The authors conducted the present study to elucidate what elements characterize the nasal profiles of patients with unilateral cleft lips (CLs). MATERIALS AND METHODS: A total of 40 Japanese unilateral CL patients were studied. For each patient, the nasal profile curve was traced on three dimensional computer tomography image. Then four points were marked on the contour. The points were NAS (Nasion), MAP (the Most Anterior Point on the nasal profile curve), GPRN (the Genuine Pronasale: the point on the nasal curve at which the curve protrudes most), and SBN (Subnasale: the point at the columellar base). Using specially designed software, the distances between these marking points were measured along the nasal profile curve. RESULTS: In CL patients, the distance between the MAP and GPRN is longer, and the GPRN is located more inferiorly than in non-cleft persons. CONCLUSION: The nasal tip tends to become round and to droop in unilateral CL patients. In order to avoid this deformity pattern, the nasal tip should be reshaped to present a sharper curvature and corrected superiorly. PMID- 18430580 TI - Intraoperative device closure of large secundum atrial septal defects; a safe alternative to transcatheter closure. AB - OBJECTIVE: The aim of this study is to report our short and mid-term results of intraoperative device closure (IODC) in large secundum atrial septal defects (ASD), to evaluate its safety and to determine the impact of 'short' rim on the results. METHODS: Sixty-eight patients with an ASD underwent IODC through a right minithoracotomy. Patients were divided into two groups: 37 patients in group I with one short rim (< or =5 mm) and 31 in group II with sufficient rims. A 2.5-3 cm parasternal incision was made in the right third or fourth intercostal space. A specially designed plastic sheath loaded with the device was inserted through the purse-string sutures placed on the right atrium. Under transesophageal echocardiographic guidance, it was advanced through the ASD into the left atrium and the device was deployed in place. RESULTS: The procedure was successful in all patients. The maximum diameter of the ASD ranged from 20 to 37 mm (mean 25+/ 5 mm). There were 16 patients with the diameter of ASD more than 30 mm. The mean size of implanted devices was 29+/-4mm. Redeployment with larger device occurred in seven patients in group I and three in group II (p>0.05). Intracardiac manipulation time was 22+/-10 min in group I and 16+/-11 min in group II (p<0.01). The total occlusion rate was 84% immediately after operation, 97% at 3 months, 98% at 1 year, and 100% at 2-, 3-, 4-year follow-up. There were no other late complications during the follow-up period of 3-63 months (mean 27+/-18 months). CONCLUSIONS: IODC is a safe and feasible technique in closing large ASDs. It has the advantages of cost savings, cosmetic results, and less trauma. Early and mid-term results are encouraging. In patients with ASD of a short rim, a larger device is recommended which does not influence the success rate of IODC. PMID- 18430581 TI - Does endoesophageal ultrasound-guided fine-needle aspiration replace mediastinoscopy in mediastinal staging of thoracic malignancies? AB - OBJECTIVE: To determine the impact of endoesophageal ultrasound-guided fine needle aspiration (EUS-FNA) on management of thoracic malignancies. METHODS: One hundred and twenty patients referred for invasive diagnostic and resection of thoracic malignancies were studied prospectively. Negative and inconclusive EUS FNA findings were assessed by video-assisted mediastinoscopic lymphadenectomy (VAMLA) or open lymphadenectomy. RESULTS: One hundred and twenty patients, aged 64.1 years (range 38-85) underwent 120 EUS-FNA, 53 video-assisted mediastinoscopic and 48 open lymphadenectomies for diagnosis and treatment of 99 lung carcinoma, six lung metastases, five mesothelioma, three lymphoma, and eight other conditions. EUS-FNA showed T4 in 15/120 and adrenal or hepatic metastases in 9/120 cases. Prevalence of mediastinal lymph node metastases was 51.7%. EUS FNA false-negative rate was 25.3%. EUS-FNA sensitivity was 91.7%, 78.1% and 43.8% for bulky disease, enlarged mediastinal nodes or normal nodes on CT scan, 50% and 96.6% for right- and left-sided tumours, and 80.6%, 78.9%, 23.8% and 25.0% for the lymph node stations 7, 5/6, 4R, and 4L. A 38.3% respectively 100% cut-down of mediastinoscopies leads in 7.5% respectively 20.8% to incorrect treatment decisions. CONCLUSIONS: EUS-FNA sensitivity depends on the localisation of the primary tumour, and extent and location of mediastinal disease. For left-sided tumours, EUS-FNA improves mediastinal staging by assessing stations 5 and 6 inaccessible to conventional mediastinoscopy. For extended mediastinal disease, mediastinoscopy can be avoided or spared for restaging after neoadjuvant therapy. Exclusion of mediastinal involvement requires mediastinoscopy or open lymphadenectomy. Beyond mediastinal nodal staging, EUS-FNA may detect T4 and M1 situations. Thus, EUS-FNA is a useful supplement to and not the replacement of mediastinoscopy. PMID- 18430582 TI - Spectroscopic analysis of the interaction between gallium(III) and apoovotransferrin. AB - Ovotransferrin is a main member of transferrin family and has a dual role in both the transport of iron and antibacterial function. Gallium-67 is widely used as an imaging agent for tumors. It has been reported that Ga(3+) can bind to apoovotransferrin at two sites, one in the N-terminal lobe and another in the C terminal lobe. However, several details of the interaction between Ga(3+) and apoOTf remain unclear. Here, we report detailed investigations into the interactions of Ga(3+) with apoovotransferrin at the molecular level. First, the characteristics of Ga(3+) binding to apoovotransferrin were analyzed using UV difference spectra. The results show that Ga(3+) prefers to bind to the N terminal site rather than the C-terminal site under the experimental conditions. Effective stability constants of logK(N)=18.88+/-0.24 and logK(C)=17.65+/-0.12 were determined. Second, conformational changes in apoovotransferrin during Ga(3+) binding were studied using 2-p-toluidinylnaphthalene-6-sulfonate (TNS) as a fluorescence probe. Apoovotransferrin undergoes a large conformational change when Ga(3+) binds to the N-terminal site, and a smaller conformational change when the ion binds to the C-terminal site. UV difference spectra were also used to measure the rate at which EDTA removes Ga(3+) from ovotransferrin carrying one Ga(3+) at the N-terminal site. Ga(3+) removal from the N-terminal binding site follows simple saturation kinetics. PMID- 18430583 TI - Epigenetic biomarkers for human cancer: the time is now. AB - The importance of epigenetic processes in the development of cancer is clear. The study of epigenetics is therefore bound to contribute to the improvement of human health. Aberrations in DNA methylation, post-translational modifications of histones, chromatin remodeling and microRNAs patterns are the main epigenetic alterations, and these are associated with tumorigenesis. Epigenetic technologies in cancer studies are helping increase the number of cancer candidate genes and allow us to examine changes in 5-methylcytosine DNA and histone modifications at a genome-wide level. In fact, all the various cellular pathways contributing to the neoplastic phenotype are affected by epigenetic genes in cancer. They are being explored as biomarkers in clinical use for early detection of disease, tumor classification and response to treatment with classical chemotherapy agents, target compounds and epigenetic drugs. Encouraging results have been obtained with histone deacetylase and DNA methyltransferase inhibitors, leading the US Food and Drug Administration to approve several of them for the treatment of hematological malignancies and lymphoproliferative disorders, such as myelodysplastic syndrome and cutaneous lymphoma. However, many tasks remains to be done, such as the clinical validation of epigenetic biomarkers to allow the accurate prediction of the outcome of cancer patients and their potential chemosensitivity to current pharmacological treatments. PMID- 18430584 TI - Study on the photocatalytic degradation of methyl orange in water using Ag/ZnO as catalyst by liquid chromatography electrospray ionization ion-trap mass spectrometry. AB - A nanocrystal catalyst Ag/ZnO was successfully synthesized using a simple solvothermal method in this study. This catalyst was characterized by X-ray diffraction (XRD) and transmission electron microscopy (TEM). The results showed that this catalyst was composed of metallic Ag and ZnO. The photodegradation of methyl orange (MO) was investigated in aqueous suspension containing Ag/ZnO catalyst under UV irradiation. Liquid chromatography coupled with electrospray ionization ion-trap mass spectrometry was applied to the analysis of the samples coming from the photocatalytic degradation of MO. The experimental results showed that there were four intermediates existing in the photocatalytic reaction. MO could be mineralized in the Ag/ZnO suspension after 60 min illumination. PMID- 18430585 TI - Recombinant human epidermal growth factor inclusion body solubilization and refolding at large scale using expanded-bed adsorption chromatography from Escherichia coli. AB - Amongst the various endogenous growth factors, epidermal growth factor (EGF) plays an important role in normal wound healing of tissue such as skin, cornea and gastrointestinal tract. Various studies have proved that supplementing recombinant human EGF (rhEGF) results in significant augmentation of wound healing. In the present work, a high level expression system with poly-arginine sequences was used for the production of recombinant human EGF (rhEGF) as inclusion bodies. The inclusion bodies were solubilized and the protein was refolded by using expanded-bed adsorption chromatography. The renatured protein was digested with appropriate concentration of trypsin and subsequently the digested rhEGF is purified by passing through ion-exchange chromatography (Toyopearl-SP) to obtain a biologically active protein. This process is the shortest process with reduced number of steps of purification, eliminates the usage of preparative reversed phase HPLC (RP-HPLC) for final purification, which is an expensive technique. The purified protein was analyzed by RP-HPLC, showing a purity > 99% and size exclusion chromatography profile shows that there are minimal aggregates, with 99% renatured active protein. The purified rhEGF showed a specific activity of 5 x 10(5) IU/mg protein, in comparison with NIBSC standard (1st International Standard of rDNA-derived EGF, Code 91/530). The process has been successfully adopted at 100 L fermentation scale and the rhEGF based formulation has been commercialized with brand name REGEN D, with excellent clinical results. PMID- 18430586 TI - High yield purification of soluble guanylate cyclase from bovine lung. AB - Soluble guanylate cyclase (sGC), the main target of nitric oxide (NO), is a cytosolic, heme-containing, heterodimeric enzyme that catalyzes the conversion of guanosine 5'-triphosphate (GTP) to 3,5'-cyclic guanosine monophosphate (cGMP) and pyrophosphate (PPi) in the presence of Mg2+. Cyclic GMP is then involved in transmitting the NO activating signals to a variety of downstream effectors such as cyclic-nucleotide-gated channels, protein kinases, and phosphodiesterases. In this work, sGC has been purified from bovine lung. The lungs were subjected to grinding and extraction with buffer at physiological pH followed by centrifugation. The resulting solution was subjected to successive column chromatography on DEAE- and Q-Sepharose, Ceramic Hydroxyapatite, Resource Q, and GTP-agarose. The purified enzyme migrated as a two-band protein on SDS-PAGE corresponding to sGC subunits alpha (M(r)=77,532) and beta (M(r)=70,500) and had an A(280 nm)/A(430 nm) of approximately 1 indicating one heme per heterodimer. The yield of enzyme was 8-10mg from 4 to 5 kg bovine lungs. V(max) and K(m) of non-stimulated sGC were 22 nmol/mg/min and 180 microM, respectively. Upon stimulation with the NO donor 3-ethyl-3-(ethylaminoethyl)-1-hydroxy-2-oxo-1 triazene, the V(max) increased to 1330 nmol/mg/min while the K(m) dropped to 43 microM. The quality and quantity of enzyme make it suitable for studies to probe the structure and catalytic mechanism of this enzyme and for research related to drug discovery. PMID- 18430587 TI - Biophysical and biochemical characterization of recombinant human Pop2 deadenylase. AB - Pop2, a component of the Ccr4-Not complex, functions as a deadenylase both in vitro and in vivo. In this research, we found that the recombinant human Pop2 (hPop2) mainly existed in a compact monomeric state with a a + b tertiary structure type. The percentages of the secondary structures evaluated from the CD spectrum were about 37% a-helix, 14% b-sheet, and 19% b-turns. The optimal condition for hPop2 catalysis was pH 7-8 at 37 C. Mg2+, Mn2+, and Co2+ had similar effects on the deadenylation activity of hPop2, and the optimal concentration was 0.3-0.5 mM. The deadenylase activity of hPop2 was, at least partially, specific when coordinated with divalent metal ions. The enzyme was not inhibited much by the nucleotide analogs, and the product 50-AMP was the most efficient inhibitor. The dissimilarity in the metal ion dependence and inhibitory effects of the nucleotide analogs suggested that various deadenylases might have differential regulation mechanisms. PMID- 18430588 TI - Application of PCR-RF-SSCP to study major histocompatibility class II B polymorphism in common carp (Cyprinus carpio L.). AB - A variety of methods have been applied for the characterization of major histocompatibility (MH) polymorphism in fish. We optimized a technique designated polymerase chain reaction-restriction fragments-single strand conformation polymorphism (PCR-RF-SSCP) for screening a large number of individuals for the Cyca-DAB1 and Cyca-DAB2 genes polymorphism in common carp. The advantages of this technique are simplicity, high sensitivity and low costs. PCR-RF-SSCP analysis revealed different genotypes consisting of unique combinations of the Cyca-DAB1 and Cyca-DAB2 sequences with the number of SSCP bands clearly correlating with the degree of heterozygosity of the Cyca-DAB1 and Cyca-DAB2 genes. We found four alleles for Cyca-DAB1 (*02-*05) gene but only one allele for Cyca-DAB2 (*02) and noted that the Cyca-DAB2 gene was either homozygous or absent. PCR-RF-SSCP analysis of n=79 carp individuals challenged with Aeromonas hydrophila indicated that individuals bearing no Cyca-DAB2 gene showed higher cumulative mortality and lower bacterial agglutination titers during the experiment. We suggest that our PCR-RF-SSCP method can be used to study correlations of different MH class II B genotypes/alleles with resistance of common carp to specific pathogens. PMID- 18430589 TI - Activation-induced force enhancement in human adductor pollicis. AB - It has been known for a long time that the steady-state isometric force after muscle stretch is bigger than the corresponding force obtained in a purely isometric contraction for electrically stimulated and maximal voluntary contractions (MVC). Recent studies using sub-maximal voluntary contractions showed that force enhancement only occurred in a sub-group of subjects suggesting that force enhancement for sub-maximal voluntary contractions has properties different from those of electrically-induced and maximal voluntary contractions. Specifically, force enhancement for sub-maximal voluntary contractions may contain an activation-dependent component that is independent of muscle stretching. To address this hypothesis, we tested for force enhancement using (i) sub-maximal electrically-induced contractions and stretch and (ii) using various activation levels preceding an isometric reference contraction at 30% of MVC (no stretch). All tests were performed on human adductor pollicis muscles. Force enhancement following stretching was found for all subjects (n=10) and all activation levels (10%, 30%, and 60% of MVC) for electrically-induced contractions. In contrast, force enhancement at 30% of MVC, preceded by 6s of 10%, 60%, and 100% of MVC was only found in a sub-set of the subjects and only for the 60% and 100% conditions. This result suggests that there is an activation dependent force enhancement for some subjects for sub-maximal voluntary contractions. This activation-dependent force enhancement was always smaller than the stretch-induced force enhancement obtained at the corresponding activation levels. Active muscle stretching increased the force enhancement in all subjects, independent whether they showed activation dependence or not. It appears that post-activation potentiation, and the associated phosphorylation of the myosin light chains, might account for the stretch-independent force enhancement observed here. PMID- 18430590 TI - Accelerated cerebral white matter development in preterm infants: a voxel-based morphometry study with diffusion tensor MR imaging. AB - Twenty-seven preterm infants were compared to 10 full-term infants at term equivalent age using a voxel-based analysis of diffusion tensor imaging of the brain. Preterm infants exhibited higher fractional anisotropy values, which may suggest accelerated maturation, in the location of the sagittal stratum. While some earlier findings in preterm infants have suggested developmental delays, the results of this study are more consistent with accelerated white matter development, possibly as a result of increased sensorimotor stimulation in the extrauterine environment. These results are the first to suggest that the increased intensity of stimulation associated with preterm birth may advance the process of white matter maturation in the human brain. Questions remain about whether these findings reflect acceleration of the process of white matter maturation generally, or localized alterations induced specifically by activity in affected pathways. PMID- 18430591 TI - Molecular systematics of Volvocales (Chlorophyceae, Chlorophyta) based on exhaustive 18S rRNA phylogenetic analyses. AB - The taxonomy of Volvocales (Chlorophyceae, Chlorophyta) was traditionally based solely on morphological characteristics. However, because recent molecular phylogeny largely contradicts the traditional subordinal and familial classifications, no classification system has yet been established that describes the subdivision of Volvocales in a manner consistent with the phylogenetic relationships. Towards development of a natural classification system at and above the generic level, identification and sorting of hundreds of sequences based on subjective phylogenetic definitions is a significant step. We constructed an 18S rRNA gene phylogeny based on 449 volvocalean sequences collected using exhaustive BLAST searches of the GenBank database. Many chimeric sequences, which can cause fallacious phylogenetic trees, were detected and excluded during data collection. The results revealed 21 strongly supported primary clades within phylogenetically redefined Volvocales. Phylogenetic classification following PhyloCode was proposed based on the presented 18S rRNA gene phylogeny along with the results of previous combined 18S and 26S rRNA and chloroplast multigene analyses. PMID- 18430592 TI - Unique size and location of a glomus tumor with a review of the literature. PMID- 18430593 TI - Latissimus dorsi transfer to restore external rotation with reverse shoulder arthroplasty: a biomechanical study. AB - In patients with pseudoparesis of the shoulder resulting from irreparable rotator cuff tears, reverse shoulder arthroplasty (RSA) can restore active elevation, but external rotation remains less predictable. Latissimus dorsi transfer (LDT) has been shown to be effective in restoring external rotation in patients with posterosuperior tears of the rotator cuff. The aim of this study is to determine the capacity of the LDT to restore external rotation in combination with RSA and to investigate the mechanical advantage produced by 3 different insertion sites. A biomechanical model was created using a reverse total shoulder prosthesis with 3 different transfer insertions. Moment arms were measured for 2 static positions and 1 motion of the humerus. The moment arm analysis showed that LDT can improve active external rotation in the setting of a reverse prosthesis. An insertion site on the posterior side of the greater tuberosity (adjacent to the teres minor insertion) produced a greater external rotation moment arm. PMID- 18430594 TI - Latissimus dorsi transfer for irreparable rotator cuff tears: a longitudinal study. AB - Latissimus dorsi transfer is indicated for isolated posterior superior defects of the rotator cuff. Additional lesions limit the success of the outcome, but they are relatively frequent in revision surgery. We analyzed their influence on the postoperative function in 52 patients with an irreparable tear of the rotator cuff (35 primary operations, 17 revision surgeries). We observed a continuous improvement in the Constant score from 36 to 69 points, also in ROM, strength, relief of pain and of different subjective parameters for the entire group in consecutive examinations at 11.1, 35.7 and 50.2 months. We found increased osteoarthritis (from 1.0 to 1.5 mm), as well as a decrease in the acromiohumeral distance (from 5.6 to 4.7 mm). In contrast, we detected a slight decrease in the values in the revision group and in the presence of an additional subscapularis lesion. PMID- 18430596 TI - Do visual impairments affect risk of motor problems in preterm and term low birth weight adolescents? AB - BACKGROUND: Increased prevalence of motor and visual problems has been reported in low birth weight populations, but the association between them is less studied. AIM: To examine how visual impairments may be associated with the increased risk of motor problems in low birth weight adolescents. METHODS: Fifty one very low birth weight adolescents (VLBW), 56 term small for gestational age (SGA) and 75 term control adolescents, without cerebral palsy, were examined at the age of 14. Motor skills were examined by the Movement Assessment Battery for Children. Visual functions included visual acuity, contrast sensitivity, nystagmus, strabismus, stereoacuity, accommodation, convergence and visual perception (Visual-Motor Integration test). An abnormality score was calculated as the sum of visual impairments. We used odds ratio as an estimate of the relative risk of having motor problems. RESULTS: The odds of having motor problems were 10.4 (95% CI: 2.2-49.4) in the VLBW group and 5.1 (95% CI: 1.0 25.8) in the SGA group compared with the control group. The odds of having motor problems in the VLBW group were influenced by all visual variables, and most by visual acuity, when we adjusted for these separately. The greatest reduction in OR was found when adjusting for the abnormality score (adjusted OR: 6.8; 95% CI: 1.3-34.5). In the SGA group the odds of having motor problems were relatively unaffected by the visual variables and the abnormality score. CONCLUSIONS: Visual impairments influence motor problems in VLBW adolescents, whereas motor problems in SGA adolescents seem to be unaffected by visual impairments. PMID- 18430597 TI - The rabbit as a model to study asthma and other lung diseases. AB - No single animal model is able to reproduce all the features of human asthma. However, the similarities between neonatally immunised rabbits and human asthma highlight the value of this model in the investigation of asthma pathophysiology and in the development of therapeutic agents. Airway inflammation and airway responses to various stimuli including histamine, adenosine 5'monophosphte and antigen in allergic rabbits have shown similarities with the responses observed in asthmatics. Furthermore, functional studies in rabbit airways show they are poorly responsive to capsaicin as are human airways. Chronic pre-treatment with capsaicin desensitises the TRPV(1) receptor enabling studies into the effect of this drug in both rabbits and man. The allergic rabbit model has been used extensively in assessing the various classes of anti-asthma drugs and is sensitive to similar drugs as patients with asthma, including beta-adrenoceptor agonists, corticosteroids, phosphodiesterase inhibitors and theophylline. This article highlights the usefulness of the rabbit as a species to study lung biology. PMID- 18430598 TI - Isolation, characterization and cDNA sequencing of a Kazal family proteinase inhibitor from seminal plasma of turkey (Meleagris gallopavo). AB - The turkey reproductive tract and seminal plasma contain a serine proteinase inhibitor that seems to be unique for the reproductive tract. Our experimental objective was to isolate, characterize and cDNA sequence the Kazal family proteinase inhibitor from turkey seminal plasma and testis. Seminal plasma contains two forms of a Kazal family inhibitor: virgin (Ia) represented by an inhibitor of moderate electrophoretic migration rate (present also in the testis) and modified (Ib, a split peptide bond) represented by an inhibitor with a fast migration rate. The inhibitor from the seminal plasma was purified by affinity, ion-exchange and reverse phase chromatography. The testis inhibitor was purified by affinity and ion-exchange chromatography. N-terminal Edman sequencing of the two seminal plasma inhibitors and testis inhibitor were identical. This sequence was used to construct primers and obtain a cDNA sequence from the testis. Analysis of a cDNA sequence indicated that turkey proteinase inhibitor belongs to Kazal family inhibitors (pancreatic secretory trypsin inhibitors, mammalian acrosin inhibitors) and caltrin. The turkey seminal plasma Kazal inhibitor belongs to low molecular mass inhibitors and is characterized by a high value of the equilibrium association constant for inhibitor/trypsin complexes. PMID- 18430599 TI - Nucleoside composition of Heloderma venoms. AB - Venoms of Heloderma horridum and Heloderma suspectum were analyzed for the possible presence of purine and pyrimidine nucleosides. Adenosine, cytidine, guanosine, hypoxanthine, inosine, and uridine were found in mug quantities. These amounts are much smaller than those seen in many elapid or viperine venoms, but greater and more varied than those found in crotaline venoms. While their contribution to the hypotension induced by Heloderma venoms may be minor, venom nucleosides nonetheless act in concert with kallikreins/hemorrhagins, alkaline phosphomonoesterase, 5'-nucleotidase, helodermin, helospectins, helothermine, and serotonin. The use of nucleosides as toxins is therefore a generalized squamate strategy, rather than the exclusive province of snakes. Both Heloderma venoms were found to be devoid of NADase and phosphodiesterase activities. Enzymes to release endogenous purines in the prey, are not significant components of Heloderma venoms. PMID- 18430600 TI - A compact linac for intensity modulated proton therapy based on a dielectric wall accelerator. AB - A novel compact CT-guided intensity modulated proton radiotherapy (IMPT) system is described. The system is being designed to deliver fast IMPT so that larger target volumes and motion management can be accomplished. The system will be ideal for large and complex target volumes in young patients. The basis of the design is the dielectric wall accelerator (DWA) system being developed at the Lawrence Livermore National Laboratory (LLNL). The DWA uses fast switched high voltage transmission lines to generate pulsed electric fields on the inside of a high gradient insulating (HGI) acceleration tube. High electric field gradients are achieved by the use of alternating insulators and conductors and short pulse times. The system will produce individual pulses that can be varied in intensity, energy and spot width. The IMPT planning system will optimize delivery characteristics. The system will be capable of being sited in a conventional linac vault and provide intensity modulated rotational therapy. Feasibility tests of an optimization system for selecting the position, energy, intensity and spot size for a collection of spots comprising the treatment are underway. A prototype is being designed and concept designs of the envelope and environmental needs of the unit are beginning. The status of the developmental new technologies that make the compact system possible will be reviewed. These include, high gradient vacuum insulators, solid dielectric materials, SiC photoconductive switches and compact proton sources. PMID- 18430601 TI - The EU optimal blood use project. AB - The EU optimal blood use project (EUOBUP) is co-funded by the European Commission and led by the Scottish National Blood Transfusion Service (SNBTS). Its purpose is to develop, evaluate and disseminate a manual that provides practical guidance and support for those seeking to improve the safety of the clinical transfusion process and the effectiveness of the prescribing of blood components. We define the optimal use of blood components as the safe, clinically effective and efficient use of the scarce resource of donated human blood. The project will build on the experience of a pilot project in optimal use of blood in the national health service in Scotland. This pilot developed training resources in the safe and effective use of blood and delivered training to a large number of practitioners. It has also developed systems to provide hospitals with comparative information on their use of blood components for specific clinical groups of patients to assist them in reviewing practice against that of their peers. PMID- 18430602 TI - Evidence for indications of fresh frozen plasma. AB - There continues to be a general but unfounded enthusiasm for fresh frozen plasma (FFP) usage across a range of clinical specialties in hospital practice. Clinical use of plasma has grown steadily over the last two decades in many countries. In England and Wales, there has not been a significant reduction in the use of FFP over the last few years, unlike red cells. There is also evidence of variation in usage among countries--use in England and Wales may be proportionately less per patient than current levels of usage in other European countries and the United States. Plasma for transfusion is most often used where there is abnormal coagulation screening tests, either therapeutically in the face of bleeding, or prophylactically in non-bleeding subjects prior to invasive procedures or surgery. Little evidence exists to inform best therapeutic plasma transfusion practice. Most studies have described plasma use in a prophylactic setting, in which laboratory abnormalities of coagulation tests are considered a predictive risk factor for bleeding prior to invasive procedures. The strongest randomised controlled trial (RCT) evidence indicates that prophylactic plasma for transfusion is not effective across a range of different clinical settings and this is supported by data from non-randomised studies in patients with mild to moderate abnormalities in coagulation tests. There are also uncertainties whether plasma consistently improves the laboratory results for patients with mild to moderate abnormalities in coagulation tests. There is a need to undertake new trials evaluating the efficacy and adverse effects of plasma, both in bleeding and non-bleeding patients, to understand whether the "presumed" benefits outweigh the "real risks". In addition, new haemostatic tests should be validated which better define risk of bleeding. PMID- 18430603 TI - How the walls come crumbling down: recent structural biochemistry of plant polysaccharide degradation. AB - The recent years have witnessed considerable developments in the interpretation of the three-dimensional structures of plant polysaccharide-degrading enzymes in the context of their functional specificity. A plethora of new structures of catalytic, carbohydrate-binding and protein-scaffolding modules involved in (hemi)cellulose catabolism has emerged in harness with sophisticated biochemical analysis. Despite significant advances, a full understanding of the intricacies of substrate recognition and catalysis by these diverse and specialised enzymes remains an important goal, especially if the application potential of these biocatalysts is to be fully realised. PMID- 18430604 TI - Evaluation of DNA damage and mutagenicity induced by lead in tobacco plants. AB - Tobacco (Nicotiana tabacum L. var. xanthi) seedlings were treated with aqueous solutions of lead nitrate (Pb2+) at concentrations ranging from 0.4 mM to 2.4 mM for 24 h and from 25 microM to 200 microM for 7 days. The DNA damage measured by the comet assay was high in the root nuclei, but in the leaf nuclei a slight but significant increase in DNA damage could be demonstrated only after a 7-day treatment with 200 microM Pb2+. In tobacco plants growing for 6 weeks in soil polluted with Pb2+ severe toxic effects, expressed by the decrease in leaf area, and a slight but significant increase in DNA damage were observed. The tobacco plants with increased levels of DNA damage were severely injured and showed stunted growth, distorted leaves and brown root tips. The frequency of somatic mutations in tobacco plants growing in the Pb2+-polluted soil did not significantly increase. Analytical studies by inductively coupled plasma optical emission spectrometry demonstrate that after a 24-h treatment of tobacco with 2.4 mM Pb2+, the accumulation of the heavy metal is 40-fold higher in the roots than in the above-ground biomass. Low Pb2+ accumulation in the above-ground parts may explain the lower levels or the absence of Pb2+-induced DNA damage in leaves. PMID- 18430605 TI - Mixed complexes of alkaline earth uranyl carbonates: a laser-induced time resolved fluorescence spectroscopic study. AB - The interaction of the alkaline earth ions Mg(2+), Sr(2+) and Ba(2+) with the uranyl tricarbonato complex has been studied by time-resolved laser-induced fluorescence spectroscopy. In contrast to the non-luminescent uranyl tricarbonato complex at ambient temperature the formed products show luminescence properties. These have been used to determine the stoichiometry and complex stabilities of the formed compounds. As the alkaline earth elements are located in an outer shell of the complex the influence of the type of the alkaline earth element on the stability constant is not very drastic. The stability constants range from log beta113 degrees = 26.07+/-0.13 to log beta113 degrees = 26.93+/-0.25 for the first reaction step and from log beta213 degrees = 29.73+/-0.47 to log beta213 degrees = 30.79+/-0.29 for the overall complex formation with two alkaline earth ions. PMID- 18430606 TI - Effect of atorvastatin on plasma levels of asymmetric dimethylarginine in patients with non-ischaemic heart failure. AB - BACKGROUND: Elevated plasma levels of asymmetric dimethylarginine (ADMA), an endothelial nitric oxide synthase (eNOS) inhibitor, may contribute to endothelial dysfunction in chronic heart failure (CHF). Since statins upregulate eNOS and ameliorate endothelial dysfunction in non-ischaemic CHF, we hypothesized that this may be in part through modification of ADMA. AIM: To evaluate the effect of atorvastatin on the relationship between ADMA and endothelial function in non ischaemic CHF. METHODS: Twenty-four patients with CHF (ejection fraction <40%, New York Heart Association Functional Classes II and III) were randomised to atorvastatin treatment (40 mg) or placebo once daily for 6 weeks in a double blinded, placebo-controlled crossover study. Plasma ADMA and l-arginine levels were measured by HPLC. Endothelial function was assessed by flow-mediated dilatation and invasive forearm plethysmography. RESULTS: Post-statin therapy, endothelial function was improved (p<0.05) independent of LDL-cholesterol reductions, but no changes were observed in ADMA levels or the l-arginine to ADMA ratio. There was a trend for ADMA to inversely correlate with endothelial function at baseline. CONCLUSIONS: Short-term atorvastatin treatment in non ischaemic CHF improves endothelial function but has no effect on ADMA or the l arginine to ADMA ratio. Our finding suggests that the observed statin-induced improvements in endothelial function are likely mediated via alternative pathways. PMID- 18430608 TI - Homology modeling, agonist binding site identification, and docking in octopamine receptor of Periplaneta americana. AB - AY333178 (from Periplaneta americana, 628 AAs) was selected as a target octopamine receptor (OAR) class OAR2 for this study using Discovery Studio (DS Modeling1.1/1.2, Accelrys Inc.). Blast similarity search was performed and identified that AY333178 contains N-terminal domain of GPCR. Based upon Blast and Pfam results, Rhodopsin 1U19 (protein data bank) was considered as an ideal homologue and used as a template for homology modeling due to its higher X-ray resolution at 2.2A. Sequence alignment between AY333178 and 1U19 was done using Align123 followed by a manual modification. The final alignment was carefully evaluated and evidenced to be matching the conserved residue data for class A GPCR fairly well. The 3D model of AY333178 was generated with MODELER, and further refined using CHARMm. Superimposition of the model was done over the template 1U19. Two fairly consistent profiles were observed demonstrating AY333178 model was reasonable and could be employed for the further docking study. Agonist docking into OAR2 model was done using LigandFit. The superimposition of two top poses of representative agonists was performed with a soft surface generated. Those models are considered to be used in designing new leads for hopefully more active compounds. Further research on the comparison of models for the agonists may elucidate the mechanisms of OAR2-ligand interactions. PMID- 18430609 TI - Automatic perceptual color map generation for realistic volume visualization. AB - Advances in computed tomography imaging technology and inexpensive high performance computer graphics hardware are making high-resolution, full color (24 bit) volume visualizations commonplace. However, many of the color maps used in volume rendering provide questionable value in knowledge representation and are non-perceptual thus biasing data analysis or even obscuring information. These drawbacks, coupled with our need for realistic anatomical volume rendering for teaching and surgical planning, has motivated us to explore the auto-generation of color maps that combine natural colorization with the perceptual discriminating capacity of grayscale. As evidenced by the examples shown that have been created by the algorithm described, the merging of perceptually accurate and realistically colorized virtual anatomy appears to insightfully interpret and impartially enhance volume rendered patient data. PMID- 18430610 TI - Pseudoachalasia caused by retroperitoneal B-cell lymphoma. PMID- 18430611 TI - Liposomal delivery improves the growth-inhibitory and apoptotic activity of low doses of gemcitabine in multiple myeloma cancer cells. AB - Gemcitabine-loaded pegylated unilamellar liposomes (200 nm) were proposed for the treatment of multiple myeloma cancer disease. Physicochemical and technological parameters of liposomes were evaluated by using laser light scattering and gel permeation chromatography. The growth-inhibitory activity of gemcitabine-loaded liposomes compared to the free drug was assayed in vitro on U266 (autocrine, interleukin-6-independent) and INA-6 (IL-6-dependent) multiple myeloma cell lines. Liposomes noticeably improved the growth-inhibitory activity of gemcitabine in terms of both dose-dependent and incubation-time effects. Liposomal delivery of gemcitabine consistently and significantly increased induction of apoptosis and caused a complete inhibition of proliferation. Liposomes were able to interact with multiple myeloma cells as demonstrated by confocal laser scanning microscopy and hence to improve the intracellular gemcitabine delivery. Gemcitabine-loaded liposomes were much more effective in vitro than the free drug. This formulation may offer even more in vivo advantages both in terms of drug pharmacokinetic and biodistribution. PMID- 18430613 TI - Anatomical and physiological properties of pelvic ganglion neurons in female mice. AB - Most neurons that regulate motility and blood flow in female pelvic organs are located within pelvic (paracervical) ganglia. In this study we investigated the anatomical and physiological properties of neurons within mouse (C57/Bl/6) paracervical ganglia. Most neurons showed immunoreactivity for choline acetyl transferase (CHAT) and were presumably cholinergic. Few neurons (approximately 5%) were tyrosine hydroxylase (TH) positive. Immunohistochemical labelling for microtubule associated protein 2 showed most neurons had small somata (cross sectional area approximately 300 microm(2)) and lacked dendrites. Action potential (AP) discharge characteristics, determined by depolarising current step injection, revealed most neurons (70%) adapted rapidly to depolarising current injection and were classified as "phasic". The remaining neurons discharged APs throughout the current step and were classified as "tonic". Membrane properties and current-voltage relationships were similar in phasic and tonic neurons, however the afterhyperpolarisation was significantly smaller in tonic neurons. Stimulation of preganglionic axons usually evoked a single strong preganglionic input (21/27 and 9/10 for pelvic and hypogastric nerves, respectively). In 19 preparations where we tested for inputs from both nerves pelvic inputs predominated (23/45 neurons) and inputs via the hypogastric nerve were rarely observed (3/45 neurons). Together, our data indicate that most neurons within mouse paracervical ganglia are cholinergic and parasympathetic. As there is little anatomical or functional evidence for integration of preganglionic inputs we propose that the role of paracervical neurons is restricted to one of spatial amplification or filtering of preganglionic inputs. PMID- 18430612 TI - Regulation of cardiac innervation and function via the p75 neurotrophin receptor. AB - Homeostatic regulation of cardiac function is dependent on the balance of inputs from the sympathetic and parasympathetic nervous systems. We investigated whether the p75 neurotrophin receptor plays a developmental role in cardiac innervation by analyzing sympathetic and parasympathetic fibers in the atria of p75 knockout and wildtype mice at several stages of postnatal development, and examining the effect on control of heart rate. We found that parasympathetic innervation of the atria in p75-/- mice was similar to wildtype at all time points, but that the density of sympathetic innervation was dynamically regulated. Compared to wildtype mice, the p75-/- mice had less innervation at postnatal day 4, an increase at day 28, and decreased innervation in adult mice. These changes reflect defects in initial fiber in-growth and the timing of the normal developmental decrease in sympathetic innervation density in the atria. Thus, p75 regulates both the growth and stability of cardiac sympathetic fibers. The distribution of sympathetic fibers was also altered, so that many regions lacked innervation. Basal heart rate was depressed in adult p75-/- mice, and these mice exhibited a diminished heart rate response to restraint stress. This resulted from the lack of sympathetic innervation rather than increased parasympathetic transmission or a direct effect of p75 in cardiac cells. Norepinephrine was elevated in p75-/- atria, but stimulating norepinephrine release with tyramine produced less tachycardia in p75-/- mice than wild type mice. This suggests that altered density and distribution of sympathetic fibers in p75-/- atria impairs the control of heart rate. PMID- 18430614 TI - Tracheal compression delays alveolar collapse during deep diving in marine mammals. AB - Marine mammals have very compliant alveoli and stiff upper airways; an adaptation that allows air to move from the alveoli into the upper airways, during breath hold diving. Alveolar collapse is thought occur between 30 and 100 m and studies that have attempted to estimate gas exchange at depth have used the simplifying assumption that gas exchange ceases abruptly at the alveolar collapse depth. Here we develop a mathematical model that uses compliance values for the alveoli and upper airspaces, estimated from the literature, to predict volumes of the respiratory system at depth. Any compressibility of the upper airways decreases the volume to contain alveolar air yielding lung collapse pressures 2x that calculated assuming an incompressible upper airway. A simple relationship with alveolar volume was used to predict relative pulmonary shunt at depth. The results from our model agree with empirical data on gas absorption at depth as well as the degree of tracheal compression in forced and free diving mammals. PMID- 18430615 TI - Effect of erythromycin on bronchial hyperreactivity and inflammation in ovalbumin sensitized brown Norway rats. AB - To evaluate the effect of erythromycin on bronchial hyperreactivity, inflammation, and T-cell related cytokine mRNA expression in rats sensitized to ovalbumin, three experimental groups of 10 brown Norway rats each were sensitized by breathing aerosolized ovalbumin. From day 1 to day 15, one group was given oral erythromycin 80 mg/kg/day, another group oral erythromycin 20mg/kg/day, and the third group oral saline only. A fourth control group of 10 rats breathed aerosolized saline. After sensitization, the three experimental groups were provoked by breathing ovalbumin, with the controls again breathing saline. The rats were then anesthetized and paralyzed, and pulmonary function tests were performed at baseline and after varying doses of acetylcholine. Bronchoalveolar lavage (BAL) fluid and lung tissues were examined for expression of mRNA for T cell cytokines. Our results showed that erythromycin had no beneficial effects on pulmonary function and lung inflammation in the two erythromycin-treated experimental groups compared with the saline experimental group. Th2-related cytokines and their mRNA expression in the three experimental groups were higher than in controls but did not differ among the experimental groups. In conclusion, erythromycin does not prevent bronchial hyperreactivity or an inflammatory response in ovalbumin-sensitized rats. PMID- 18430616 TI - A sensitive HPLC-MS-MS method for the determination of raltegravir in human plasma. AB - This work describes an assay system that has been developed to quantify raltegravir concentrations in human plasma using a liquid-liquid extraction technique paired with HPLC separation and MS-MS detection. The dynamic range of this assay extends from 1 to 3000 ng/mL, with a coefficient of determination (r(2), mean+/-SD) of 0.9992+/-0.0002. The mean precision values for calibration standards ranged from 0.6% to 3.0%, while accuracy values were 96.5-104.3%. This procedure is an accurate, precise, and sensitive method for raltegravir quantitation and was successfully validated using external proficiency testing. PMID- 18430617 TI - Recent advances in the management of Crohn's disease. AB - In the last 10 years many advances have been achieved in the treatment of patients with Crohn's disease, particularly in the field of biological agents. Infliximab, a tumour necrosis factor alpha antagonist, has been recently added to the therapeutic armamentarium for Crohn's disease and has greatly improved our treatment options. Infliximab has demonstrated efficacy in the induction and maintenance of remission in luminal and fistulizing Crohn's disease both in adults and children. However, the potential development of autoantibodies and the risk of serious adverse events limit the possibility of a wider use of infliximab. Searching for less immunogenicity and higher effectiveness in the last years a number of biological agents have been developed. Adalimumab, a fully human monoclonal antibody anti tumour necrosis factor alpha, has resulted effective and safe in patients with Crohn's disease, both naive and refractory to infliximab, presenting also the advantage of subcutaneous way of administration. Natalizumab also showed promising results in terms of efficacy but its safety is still under investigation. To date no particular advances have been recently appeared in the literature concerning conventional immunosuppressive drugs. Surgery remains a valid resort for refractory patients. Autologous stem cell transplantation represents a new hope as rescue treatment for patients with severe refractory Crohn's disease. PMID- 18430618 TI - Video capsule endoscopy and intestinal involvement in systemic vasculitis. PMID- 18430620 TI - UV-enhanced bioactivity and cell response of micro-arc oxidized titania coatings. AB - Using ultraviolet (UV) irradiation of micro-arc oxidized (MAO) titania coating in distilled water for 0.5 and 2h, we have achieved an enhanced bioactivity and cell response to titania surface. The MAO coating appears porous and predominantly consists of nanocrystallized anatase TiO(2). Compared with the MAO coating, the UV-irradiated coatings do not exhibit any obvious change in surface roughness, morphology, grain size and phase component; however, they have more abundant basic Ti-OH groups and become more hydrophilic because the water contact angle decreases significantly from 17.9+/-0.8 degrees to 0 degrees . In simulated body fluid (SBF), bonelike apatite-forming ability is significantly stronger on the UV irradiated coatings than the MAO coating. SaOS-2 human osteoblast-like cell attachment, proliferation and alkaline phosphatase of the cell are greater on the UV-irradiated coatings relative to the MAO coating. UV irradiation of titania results in the conversion of Ti(4+) to Ti(3+) and the generation of oxygen vacancies, which could react with the absorbed water to form basic Ti-OH groups. The enhanced bioactivity and cell response of the UV-irradiated coatings are proven to result from abundant Ti-OH groups on the coating surfaces. After storing the UV-irradiated coatings in the dark for two weeks, the basic Ti-OH groups on the coatings slightly decrease in amount and can induce apatite formation after a short period of SBF immersion, and show relative long-term stability. PMID- 18430621 TI - Does preoperative pancreatic duct stenting prevent pancreatic fistula after surgery? A cohort study. AB - BACKGROUND/OBJECTIVE: Postoperative pancreatic fistula remains a major complication after pancreatic surgeries. To prevent pancreatic fistula, one of the employed management strategies is pancreatic duct stenting. The purpose of this study was to evaluate the efficacy and safety of preoperative pancreatic stenting to prevent pancreatic fistula after surgery. METHODS: Subjects comprised 18 consecutive patients who underwent pancreatic surgeries. Patients were divided into 2 groups: stenting group (n=7); and non-stenting group (n=11). Complications after stent placement were analyzed. Compared parameters between groups included background, incidence and grading of pancreatic fistula as judged by international study group of pancreatic fistula (ISGPF) criteria, duration until drain removal, and mean maximum level of drain amylase. RESULTS: Two patients displayed mild pancreatitis with high serum amylase levels after stenting. No significant differences in background or any other compared parameters to assess drainage effect were identified between stenting and non-stenting groups. Complications related to placement of the stent tube occurred in 4 patients with tube occlusion or cholestasis. CONCLUSIONS: Although drainage effect in the stenting group was compared with that in the non-stenting group, no obvious effect was obtained. This procedure seems to require further investigation on indications for postoperative drainage to decrease the incidence of pancreatic fistula. PMID- 18430622 TI - The use of blood products in perinatal medicine. AB - Fetal and neonatal medicine is a field with many new procedures and techniques. An increasing number of centres worldwide give intrauterine transfusions, which are considered to be standard-of-care treatment for severe fetal anaemia. The survival of very prematurely born neonates, in particular of a gestational age of <28 weeks, has greatly improved over the last decade but almost all these children need transfusions. Although in many cases such blood transfusions are life saving, randomized, controlled studies investigating appropriate indications, transfusion volume and type of blood product have not been performed. Most of the protocols used are expert based. PMID- 18430623 TI - Reconstruction of skin defects after resection of severe gluteal hidradenitis suppurativa with fasciocutaneous flaps. AB - BACKGROUND: Although skin grafting after resection of the hidradenitis suppurativa is frequently used, the contour after skin grafting is unsatisfactory. We studied four patients who suffered from severe gluteal hidradenitis suppurativa and needed total skin resection of the affected lesion. METHODS: Reconstruction after total skin resection was performed with fasciocutaneous (FC) flaps of three different designs based on an arterial anatomical study: an island V-Y advancement flap based on the descending branch of the inferior gluteal artery; a rotation V-Y advancement flap based on the first perforator of the deep femoral artery and the descending branch of the inferior gluteal artery; and, a bilobed flap at each lobe containing either the descending branch or medial branch of the inferior gluteal artery. The flap designs were decided from the size and the location of the skin defect. RESULTS: The contours after reconstruction were satisfactory and did not experience any recurrence of hidradenitis suppurativa for at least 1 year after operation. CONCLUSION: We showed three different designs of FC flaps to cover the gluteal skin defects based on the anatomical study. FC flaps may be chosen for reconstruction after resection of perianal hidradenitis suppurativa. PMID- 18430625 TI - Vision and the organization of behaviour. PMID- 18430624 TI - Fracture prevalence and relationship to endocrinopathy in iron overloaded patients with sickle cell disease and thalassemia. AB - Transfusional iron overload leads to gonadal failure and low bone mass in patients with thalassemia (Thal). However, gonadal failure is rarely reported in transfused patients with sickle cell disease (SCD) and the literature regarding fracture prevalence in SCD is limited. The objective of this study was to assess self-reported fracture prevalence and its relationship to endocrinopathy in transfused Thal or SCD subjects and compare to non-transfused subjects with SCD (NonTxSCD). Eligibility was based on age> or =12 years and liver iron concentration> or =10 mg/g dry wt or serum ferritin> or =2000 ng/mL (Thal or TxSCD) or for NonTxSCD, ferritin<500 ng/mL. Data were collected by patient interview and chart review at 31 clinical centers in the U.S., Canada and the U.K. 152 subjects with Thal (52% Male; 25.6+/-0.7 years), 203 subjects with TxSCD (44% Male, 24.7+/-0.9 years: Mean+/-SE), and 65 NonTxSCD (50% Male, 22.2+/-1.3 years) were enrolled. Overall, male subjects with Thal were more likely to have sustained a fracture in their lifetime (51%) compared to TxSCD (28%) or NonTxSCD (32%) (p=0.005). There was no difference in fracture prevalence among women (Thal: 26%, TxSCD 17%, NonTxSCD: 16%). Fracture was most frequently reported in the upper extremities (53.3% of all fractures) while spine and pelvic fractures were relatively common for such a young cohort: 10.6%. Though overall fracture prevalence was not distinctly different from published healthy cohorts, fewer fractures occurred during the adolescent years. In multivariate analysis, the significant predictors of fracture prevalence were Thal diagnosis (Odds Ratio: 2.3; 1.2-4.6; 95%CI), male gender (OR: 2.6; 1.5-4.5), hypothyroidism (OR: 3.3; 1.1-9.8) and age (OR: 1.1; 1.03-1.08). These data suggest that despite similar iron burden, transfused patients with Thal are at greater risk for fracture than subjects with SCD. Male subjects with Thal and hypothyroidism are at particular risk for fracture, in contrast, transfused subjects with SCD had no greater risk of fracture compared to non-transfused SCD. Though ethnic differences in fracture risk cannot be ignored, endocrinopathy is rare in TxSCD which may also provide some protection from fracture. PMID- 18430626 TI - Plasmodesmata. PMID- 18430627 TI - Orchids. PMID- 18430628 TI - Lateral asymmetry of bodily emotion expression. PMID- 18430629 TI - Action-perception mismatch in tone-deafness. PMID- 18430630 TI - FGF induces oscillations of Hes1 expression and Ras/ERK activation. PMID- 18430631 TI - Motor learning: the FoxP2 puzzle piece. AB - Mutation of the DNA-binding region of the FOXP2 protein causes an inherited language disorder. A recent study provides the first data on mice with this mutation, which exhibit deficits in motor-skill learning and abnormal properties of neural circuits that contribute to these skills. PMID- 18430632 TI - Programmed cell death: role for MazF and MrpC in Myxococcus multicellular development. AB - Programmed cell death is of ultimate importance in embryonic development of animals. Now, programmed cell death has been shown to be an integral part of a multicellular developmental program in the bacterium Myxococcus xanthus. PMID- 18430633 TI - Fly vision: neural mechanisms of motion computation. AB - A wide range of novel approaches are being used to dissect the visual system of the fly, both the neural networks of motion detection and the performance of these networks under complex natural stimulus conditions. PMID- 18430634 TI - Photosynthetic pigments: perplexing persistent prevalence of 'superfluous' pigment production. AB - Phycobilins function as light-harvesting pigments in most cyanobacteria and red algae. Although green cyanobacteria of the genus Prochlorococcus express genes encoding enzymes that direct the synthesis of phycobilins, these pigments do not appear to play a role in light harvesting in Prochlorococcus. Now, it is shown that cyanophages infecting Prochlorococcus also contain genes for phycobilin synthesizing enzymes, and these are expressed in Prochlorococcus, raising further questions as to the role of phycobilins in the host and the virus. PMID- 18430635 TI - Social cognition: LIP activity follows the leader. AB - A new study has found that, when a macaque monkey chooses where to look, activity in parietal cortex reflects anticipated reward value, whether from fluid to drink or from social information such as dominance. PMID- 18430636 TI - Organelle evolution: what's in a name? AB - Plastids are organelles derived from cyanobacterial endosymbionts and the evolutionary process that gave rise to them is well understood. Or is it? The complete genome sequence of a recently evolved photosynthetic body in Paulinella chromatophora is cause for reflection on the distinction between 'endosymbiont' and 'organelle', and how the boundaries between these terms can blur. PMID- 18430637 TI - Biological optics: circularly polarised crustaceans. AB - A novel visual capability, the detection of circularly polarised light, has been demonstrated in a mantis shrimp, a colourful crustacean from tropical reefs. PMID- 18430638 TI - Phase coding: spikes get a boost from local fields. AB - The brain exhibits coherent, long-range oscillations, and it now appears that these oscillations play a substantial role in neural coding: they can boost the information contained in action potentials by as much as 50%. PMID- 18430639 TI - Metabolic gradients: a new system for old questions. AB - Metabolic gradients are likely to be crucial to normal and abnormal development of cells and tissues. As shown by a new study, a Xenopus egg model system has great promise to illuminate quantitative measures of metabolic gradients in living cytoplasm. PMID- 18430640 TI - Rapid pivot feeding in pipefish: flow effects on prey and evaluation of simple dynamic modelling via computational fluid dynamics. AB - Most theoretical models of unsteady aquatic movement in organisms assume that including steady-state drag force and added mass approximates the hydrodynamic force exerted on an organism's body. However, animals often perform explosively quick movements where high accelerations are realized in a few milliseconds and are followed closely by rapid decelerations. For such highly unsteady movements, the accuracy of this modelling approach may be limited. This type of movement can be found during pivot feeding in pipefish that abruptly rotate their head and snout towards prey. We used computational fluid dynamics (CFD) to validate a simple analytical model of cranial rotation in pipefish. CFD simulations also allowed us to assess prey displacement by head rotation. CFD showed that the analytical model accurately calculates the forces exerted on the pipefish. Although the initial phase of acceleration changes the flow patterns during the subsequent deceleration phase, the accuracy of the analytical model was not reduced during this deceleration phase. Our analysis also showed that prey are left approximately stationary despite the quickly approaching pipefish snout. This suggests that pivot-feeding fish need little or no suction to compensate for the effects of the flow induced by cranial rotation. PMID- 18430641 TI - Is it necessary to assume an apartheid-like social structure in Early Anglo-Saxon England? AB - It has recently been argued that there was an apartheid-like social structure operating in Early Anglo-Saxon England. This was proposed in order to explain the relatively high degree of similarity between Germanic-speaking areas of northwest Europe and England. Opinions vary as to whether there was a substantial Germanic invasion or only a relatively small number arrived in Britain during this period. Contrary to the assumption of limited intermarriage made in the apartheid simulation, there is evidence that significant mixing of the British and Germanic peoples occurred, and that the early law codes, such as that of King Ine of Wessex, could have deliberately encouraged such mixing. More importantly, the simulation did not take into account any northwest European immigration that arrived both before and after the Early Anglo-Saxon period. In view of the uncertainty of the places of origin of the various Germanic peoples, and their numbers and dates of arrival, the present study adopts an alternative approach to estimate the percentage of indigenous Britons in the current British population. It was found unnecessary to introduce any special social structure among the diverse Anglo-Saxon people in order to account for the estimates of northwest European intrusion into the British population. PMID- 18430642 TI - Social transmission of nectar-robbing behaviour in bumble-bees. AB - Social transmission of acquired foraging techniques is rarely considered outside of a vertebrate context. Here, however, we show that nectar robbing by bumble bees (Bombus terrestris)-an invertebrate behaviour of considerable ecological significance-has the potential to spread through a population at the accelerated rates typical of social transmission. Nectar robbing occurs when individuals either bite through the base of a flower to 'steal' nectar (primary robbing) or use robbing holes that others have made (secondary robbing). We found that experience of foraging from robbed flowers significantly promoted the development of primary robbing in previously legitimate foragers, thus implying that the acquisition of nectar robbing by one individual will facilitate its adoption in others. Our findings suggest that the positive feedback effects of social transmission may potentially play an ecologically important role in the relationship between plants and pollinators. PMID- 18430643 TI - Availability of prey resources drives evolution of predator-prey interaction. AB - Productivity is predicted to drive the ecological and evolutionary dynamics of predator-prey interaction through changes in resource allocation between different traits. Here we report results of an evolutionary experiment where prey bacteria Serratia marcescens was exposed to predatory protozoa Tetrahymena thermophila in low- and high-resource environments for approximately 2400 prey generations. Predation generally increased prey allocation to defence and caused prey selection lines to become more diverse. On average, prey became most defensive in the high-resource environment and suffered from reduced resource use ability more in the low-resource environment. As a result, the evolution of stronger prey defence in the high-resource environment led to a strong decrease in predator-to-prey ratio. Predation increased temporal variability of populations and traits of prey. However, this destabilizing effect was less pronounced in the high-resource environment. Our results demonstrate that prey resource availability can shape the trade-off allocation of prey traits, which in turn affects multiple properties of the evolving predator-prey system. PMID- 18430644 TI - Host manipulation by parasites in the world of dead-end predators: adaptation to enhance transmission? AB - Trophically transmitted parasites often alter their intermediate host's phenotype, thereby predisposing the hosts to increased predation. This is generally considered a parasite strategy evolved to enhance transmission to the next hosts. However, the adaptive value of host manipulation is not clear as it may be associated with costs, such as increased susceptibility to predators that are unsuitable next hosts for the parasites. We examined the ratio between the benefits and costs of host manipulation for transmission success of Acanthocephalus lucii (Acanthocephala), a parasite that alters the hiding behaviour and pigmentation of its isopod hosts. We experimentally compared the susceptibility of infected and uninfected isopods to predation by perch (Perca fluvialis; definitive host of the parasite) and dragonfly larvae (dead end). We found that the parasite predisposed the isopods to predation by both predators. However, the increased predation vulnerability of the infected isopods was higher towards perch. This suggests that, despite the costs due to non-host predation, host manipulation may still be advantageous for the parasite. PMID- 18430645 TI - Heart rate modulation in bystanding geese watching social and non-social events. AB - Simply observing other individuals interacting has been shown to affect subsequent behaviour and also hormones in 'bystander' individuals. However, immediate physiological responses of an observer have been hardly investigated. Here we present results on individuals' heart rate (HR) responses during various situations, which occur regularly in a flock of greylag geese (Anser anser, e.g. agonistic encounters, vehicles passing by). We recorded simultaneously HR and behaviour of 21 semi-tame free-roaming geese, equipped with fully implanted transmitters. We considered 304 social and 81 non-social events during which the focal individuals did not respond behaviourally. Independent of the spatial distance to the event, these HR responses were significantly greater in social contexts (e.g. departing or landing geese, agonistic interactions) than in non social situations (e.g. vehicles passing by, thunder). Focal individuals showed a significantly higher maximum HR as well as a greater HR increase in response to agonistic interactions, in which the pair partner or a family member was involved, as compared with a non-affiliated goose. Also, HR was significantly higher when the bystander watched non-affiliated geese interacting, which were higher ranking than the focal. We conclude that these differences are due to different relevance of the recorded events for the focal individual, depending on the individuals involved in the observed interaction. PMID- 18430646 TI - Feeding, fecundity and lifespan in female Drosophila melanogaster. AB - Male seminal fluid proteins induce a profound remodelling of behavioural, physiological and gene signalling pathways in females of many taxa, and typically cause elevated egg production and decreased sexual receptivity. In Drosophila melanogaster, these effects can be mediated by an ejaculate 'sex peptide' (SP), which, in addition, contributes significantly to the cost of mating in females. Recent research has revealed that SP can stimulate female post-copulatory feeding, raising the possibility that the widespread female cost of mating could be due to over-feeding. In this study, we used D. melanogaster as a model to test this hypothesis. We first show that elevated post-mating feeding is dependent upon egg production and does not occur in sterile ovoD1 mutant females. This conclusion was also supported by the increase in feeding of virgin females whose egg production was experimentally elevated. We then demonstrated that sterile ovoD1 and fertile females experienced identical survival costs of mating, related to their frequency of mating and not to female feeding rate or to egg production. We conclude that female mating costs are not the result of over-feeding, but may be due to other, potentially more direct, effects of ejaculate molecules. PMID- 18430647 TI - The mechanism of sex ratio adjustment in a pollinating fig wasp. AB - Sex ratio strategies in species subject to local mate competition (LMC), and in particular their fit to quantitative theoretical predictions, provide insight into constraints upon adaptation. Pollinating fig wasps are widely used in such studies because their ecology resembles theory assumptions, but the cues used by foundresses to assess potential LMC have not previously been determined. We show that Liporrhopalum tentacularis females (foundresses) use their clutch size as a cue. First, we make use of species ecology (foundresses lay multiple clutches, with second clutches smaller than first) to show that increases in sex ratio in multi-foundress figs occur only when foundresses are oviposition site limited, i.e. that there is no direct response to foundress density. Second, we introduce a novel technique to quantify foundress oviposition sequences and show, consistent with the theoretical predictions concerning clutch size-only strategies, that they produce mainly male offspring at the start of bouts, followed by mostly females interspersed by a few males. We then discuss the implications of our findings for our understanding of the limits of the ability of natural selection to produce 'perfect' organisms, and for our understanding of when different cue use patterns evolve. PMID- 18430648 TI - You are what your mother eats: evidence for maternal preconception diet influencing foetal sex in humans. AB - Facultative adjustment of sex ratios by mothers occurs in some animals, and has been linked to resource availability. In mammals, the search for consistent patterns is complicated by variations in mating systems, social hierarchies and litter sizes. Humans have low fecundity, high maternal investment and a potentially high differential between the numbers of offspring produced by sons and daughters: these conditions should favour the evolution of facultative sex ratio variation. Yet little is known of natural mechanisms of sex allocation in humans. Here, using data from 740 British women who were unaware of their foetus's gender, we show that foetal sex is associated with maternal diet at conception. Fifty six per cent of women in the highest third of preconceptional energy intake bore boys, compared with 45% in the lowest third. Intakes during pregnancy were not associated with sex, suggesting that the foetus does not manipulate maternal diet. Our results support hypotheses predicting investment in costly male offspring when resources are plentiful. Dietary changes may therefore explain the falling proportion of male births in industrialized countries. The results are relevant to the current debate about the artificial selection of offspring sex in fertility treatment and commercial 'gender clinics'. PMID- 18430649 TI - Protein robustness promotes evolutionary innovations on large evolutionary time scales. AB - Recent laboratory experiments suggest that a molecule's ability to evolve neutrally is important for its ability to generate evolutionary innovations. In contrast to laboratory experiments, life unfolds on time-scales of billions of years. Here, we ask whether a molecule's ability to evolve neutrally-a measure of its robustness-facilitates evolutionary innovation also on these large time scales. To this end, we use protein designability, the number of sequences that can adopt a given protein structure, as an estimate of the structure's ability to evolve neutrally. Based on two complementary measures of functional diversity catalytic diversity and molecular functional diversity in gene ontology-we show that more robust proteins have a greater capacity to produce functional innovations. Significant associations among structural designability, folding rate and intrinsic disorder also exist, underlining the complex relationship of the structural factors that affect protein evolution. PMID- 18430650 TI - Inflammatory aneurysm of arcus aorta with massive hemoptysis. AB - A 72-year-old woman was admitted to the intensive care unit with a diagnosis of acute coronary syndrome. An aneurysm in the aortic arch was detected in the radiologic investigations. The aortic arch was replaced with a Dacron graft with the patient under total circulatory arrest. A pathology evaluation revealed an inflammatory aneurysm. To date, a few cases of inflammatory aneurysms of the aortic arch have been reported. Unlike the previously reported cases, the aneurysm in our case unexpectedly presented with massive hemoptysis. PMID- 18430651 TI - Septic necrosis of the odontoid apophysis and cervical spondylodiscitis from Enterococcus faecalis endocarditis: a first report. AB - We describe a 75-year-old male patient who developed a general syndrome, with a fever of 39 degrees C, weight loss, and cervical pain, during the month following a urological procedure. The presence of positive blood cultures for Enterococcus faecalis, aortic vegetations, and severe aortic regurgitation observed with echocardiogram confirmed the diagnosis of infective endocarditis (IE). Magnetic resonance imaging of the spinal cord showed significant erosion and irregularities of the odontoid apophysis, with hyperintensity of bone marrow in T2-weighted images because of edema and inflammation. These findings suggested an infective necrosis of the odontoid apophysis. Despite the common occurrence of rheumatologic manifestations in IE, with prevalence rates of 25% to 44%, spondylodiscitis is rarely observed (5%-13%). The lumbar region is the most commonly involved. We found only one other reported case of cervical spondylodiscitis. The case we describe is the first report of septic necrosis of the odontoid apophysis associated with IE. PMID- 18430653 TI - Primary cardiac rhabdomyosarcoma of the right atrium: case report. AB - Rhabdomyosarcoma is a rare malignant tumor of the heart that accounts for 20% of all primary malignant neoplasms of the heart. Symptoms vary in accordance with the location of the mass; unfortunately, by the time the patient becomes symptomatic, the tumor has already metastasized to other organs. Diagnosis is frequently obtained via transthoracic or transesophageal echocardiography and nuclear magnetic resonance imaging. Surgery is indicated for malignant cardiac neoplasms to relieve cardiac symptoms and to prolong patient survival. Subsequent postoperative chemotherapy or radiotherapy is necessary, and the long-term prognosis is poor. We present a case of a primary cardiac rhabdomyosarcoma that arose from the lateral wall of the right atrium and required implantation of a permanent cardiac pacemaker after surgery. PMID- 18430652 TI - Long-term outcomes of surgical radiofrequency ablation for atrial fibrillation in 3 groups of patients. AB - BACKGROUND: Left atrial surgical radiofrequency ablation represents an applicable and technically less demanding method for treating paroxysmal or permanent atrial fibrillation (AF) as a concomitant procedure. The aim of this study was to review the long-term outcomes of radiofrequency linear ablation for the treatment of AF limited to the left atrium in 3 groups of patients undergoing cardiac surgery. METHODS: The study population consisted of 357 consecutive patients, who were divided into 3 groups on the basis of the underlying disease: group I, 126 patients with nonischemic mitral valve disease; group II, 164 patients with coronary artery disease and aortic and/or ischemic mitral valve disease or who underwent other concomitant procedures; and group III, 67 patients with coronary artery disease only. RESULTS: Follow-up times were between 6 and 48 months (mean, 28.3 + 9.4 months). The 30-day hospital mortality rate was 2.80% (10 patients). Total mortality during the follow-up period reached 4.48% (16 patients). At discharge, 66% of group I patients, 64% of group II patients, and 69% of group in patients were in sinus rhythm. After 24 months, 60% of group I patients, 75% of group II patients, and 67% of group III patients were in sinus rhythm. A subgroup analysis of the patients with permanent AF showed that only 54% of these patients in group I, 52% in group II, and 67% in group III had a restored sinus rhythm at 24 months. Subgroup analysis also revealed that only 6 (27%) of 22 patients with a left atrium diameter >60 mm maintained a sinus rhythm during long-term follow up. Biatrial contraction was restored in 75% of the patients with a stable sinus rhythm after 6 months of follow-up. Seven patients (2%) with symptomatic postoperative recurrent atrial arrhythmias underwent subsequent catheter ablation. CONCLUSION: Left atrial surgical radiofrequency ablation represents an applicable and technically less demanding method for treating paroxysmal or permanent AF as a concomitant procedure. Our results demonstrate the feasibility of this procedure for paroxysmal and persistent AF, with minimal risks to the patient. For permanent AF, further investigation and extensive intervention are essential. PMID- 18430654 TI - Conservative surgical treatment for active infective tricuspid valve endocarditis according to the "clover technique". AB - AIMS: This prospective study was undertaken to analyze the outcomes of conservative surgery with the "clover technique" for active infective tricuspid valve endocarditis. METHODS: Five consecutive patients underwent surgery for active infective tricuspid valve endocarditis. The mean age was 36.6 years. Four of the patients were men. In all patients, the tricuspid valve had become mutilated and infected. One patient had associated mitral endocarditis, and one had aortic endocarditis. Staphylococcus aureus was the most common bacterial species. Conservative surgery was indicated in all patients with infection limited to the leaflets and/or subvalvular apparatus of the tricuspid valve. Total resection of infected tissues was achieved in all cases. The tricuspid valve was then reconstructed according to the clover technique. A tricuspid annular ring was used in 2 patients. RESULTS: All 5 patients survived surgery. Intraoperative transesophageal and predischarge transthoracic echocardiographic evaluations showed good results in all patients. The mean follow-up time was 26.4 +/- 12.5 months. No recurrent bacterial tricuspid endocarditis occurred during follow-up. All patients were in New York Heart Association functional class I. A transthoracic echocardiography evaluation at the latest control examination showed trivial leakage (3 patients) or no residual regurgitation (2 patients); no transvalvular gradient was found in any of the patients. No tricuspid valve calcification has been detected to date. Cardiac magnetic resonance imaging analyses showed no postoperative void flow and confirmed the preservation of right ventricular function and thus the reliability of this technique. CONCLUSIONS: This novel technique is indicated for tricuspid valve endocarditis and should be considered as an adequate approach in cases of uncontrollable infection involving the tricuspid valve that is responsible for extended valve destruction. PMID- 18430655 TI - An air gun pellet retained in the heart: a case report. AB - We report the rare case of an air gun pellet retained within the myocardium. The pellet passed through the right ventricle and the interventricular septum and was retained in the posterior left ventricular wall. The patient presented with cardiac tamponade requiring urgent surgical treatment. The case report is followed by a review of the pertinent literature. PMID- 18430656 TI - Delayed cardiac tamponade after the Nuss procedure for pectus excavatum: a case report and simple management. AB - A 22-year-old man presented at our emergency department with clinical signs of cardiac tamponade that required emergency surgery. We treated this patient by using emergency echocardiographically guided pericardiocentesis with insertion of a central venous catheter for drainage without removal of the steel bar. The causes of his pericardial effusion appeared to have been displacement of the bar from a pectus excavatum repair 6 months previously and his ingestion of ginseng extract. This case illustrates a possible cause of cardiac tamponade after the Nuss procedure and a simple way to manage it. PMID- 18430657 TI - Results of adjunctive coronary endarterectomy in 548 patients. AB - Coronary endarterectomy is a controversial procedure that plays a particular role in the treatment of coronary artery disease. We retrospectively investigated the results for 548 patients who underwent coronary endarterectomy as an adjunctive therapy for coronary artery bypass graft surgery during the period between 1996 and 2004. We assessed short-term outcomes and identified risk factors for adverse outcomes. Mean patient age was 67.9 + 9.3 years and mean angina class was 2.7 + 0.3. The mean number of distal anastomoses was 3.8 + 1.1 patients (73.4%) had single and 151 (27.6%) multiple coronary artery endarterectomies. Of the 151 patients who underwent multiple endarterectomies, 97 (17.7%) had endarterectomies in 2 coronary arteries, 40 (7.2%) in 3 coronary arteries, 11 (2%) in 4 coronary arteries, 2 (0.36%) in 5 coronary arteries, and 1 (0.18%) in 6 coronary arteries. Postoperative mortality was 6.2% (34 patients). The predictors for early mortality were recent myocardial infarction and left ventricular dysfunction. Our results suggest that adjunctive coronary endarterectomy can be accomplished with acceptable results but with higher mortality rates than ordinary coronary artery bypass grafting. Adjunctive coronary endarterectomy should be considered as a last option for the surgical treatment of diffuse coronary disease. PMID- 18430658 TI - Vettath's technique of long mammary patch reconstruction of a diffusely diseased left anterior descending coronary artery without endarterectomy on the beating heart. AB - Surgery for diffuse coronary artery disease requires coronary reconstruction with or without endarterectomy. Considering the immediate and late postoperative problems of coronary endarterectomy, many surgeons now prefer coronary reconstruction without endarterectomy. Patch reconstruction of the diseased coronary artery with either the internal mammary artery or another conduit is an attractive option. This article describes the technique of long mammary patch reconstruction of the left anterior descending coronary artery without endarterectomy as an off-pump procedure. PMID- 18430659 TI - Diverticulum of the left ventricle: etiology and surgical treatment. AB - Congenital left ventricular (LV) diverticula are rare findings, particularly when first diagnosed in adulthood. We describe successful surgical repair of an isolated congenital apical LV diverticulum associated with an abnormal submitral apparatus in a young adult who received his diagnosis following a peripheral embolism. We intraoperatively identified an abnormally originating belly of the posteromedial papillary muscle, which arose from the interventricular septum, causing partition of the LV cavity and forming an apical aneurysmal communicating chamber. This anatomy of abnormal papillary muscle bifurcation was possibly the mechanism for the development of the apical diverticulum and to the best of our knowledge has not previously been described. Reattachment of the abnormal belly to its normal position preserved mitral valve competence, and an endoventricular patch repair restored LV volume and geometry. PMID- 18430660 TI - Is left ventriculotomy feasible for muscular ventricular septal defects in infants? AB - Optimal management of muscular ventricular septal defects (MVSDs) remains controversial. Left ventriculotomy is the cornerstone of surgical repair but is frequently complicated by residual shunts, left ventricular dysfunction, apical aneurysm, or arrhythmias. In this study, we evaluated the long-term outcomes of surgical repairs in infants with isolated MVSDs. We retrospectively analyzed clinical data from 56 children with MVSDs (31 males, 25 females). Follow-up by questionnaire and Doppler echocardiography was performed at discharge and between 2 and 124 months after surgery. Patient age was 2 to 40 months (median, 21 months) and weight was 3.0 to 15.3 kg (median, 5.3 kg). Two patients died after surgery (hospital mortality, 3.57%). One patient with MVSDs died of low cardiac output caused by the long duration of cardiopulmonary bypass. Another patient with Swiss cheese MVSD received a single patch closure but died of low cardiac output immediately after cardiopulmonary bypass. Immediate complications such as a third-degree atrial-ventriclar block occurred in 2 patients, but they recovered before discharge and showed no residual shunt. No deaths occurred during follow up, but a residual shunt was found in 1 patient. Delayed complete heart block requiring a pacemaker occurred in 1 patient. One patient showed paroxysmal supraventricular tachycardia that was treated with amiodarone. The left ventricular ejection fraction was 0.45-0.55 in 8 patients and 0.55-0.73 in 46 patients. No apical aneurysm was found. All the surviving patients returned to normal school life. Our results indicate that surgery is a suitable treatment option in infants and children with isolated MVSDs and that preoperative diagnosis is crucial to a successful outcome. Infants can tolerate a left ventriculotomy incision for MVSDs in the lower or apical ventricular septum. PMID- 18430661 TI - How I do it: high-quality intraoperative fluorescence imaging in off-pump coronary artery bypass grafting. AB - We have developed a simple technique for establishing high-quality intraoperative fluorescence imaging in off-pump coronary artery bypass grafting. The technique of transaortic injection of indocyanine green is an effective method of achieving clear fluorescence imaging and for evaluating the quality of graft anastomoses. We consider the images obtained with this technique to be equivalent to those obtained by conventional coronary angiography with selective enhancement of the graft. PMID- 18430662 TI - Plasma brain natriuretic Peptide after isolated on-pump coronary artery bypass grafting: prediction of postoperative adverse outcomes. AB - BACKGROUND: The heart is an endocrine organ that synthesizes 2 different natriuretic peptides: atrial natriuretic peptide and brain natriuretic peptide (BNP). We assessed the relationship between preoperative BNP levels and postoperative complications and outcomes in patients who undergo isolated coronary artery bypass grafting (CABG). METHOD AND RESULTS: The study consisted of 85 patients undergoing first-time elective CABG. Preoperative BNP levels were significantly correlated with the preoperative ejection fraction (P = .004), the number of vessels grafted (P = .016), cross-clamp time (P = .041), and perfusion time (P = .032). Preoperative BNP levels were higher in patients who developed postoperative new-onset atrial fibrillation (AF) (median BNP, 197 pg/mL versus 65 pg/mL; P = .006), in patients requiring inotropic support (189 pg/mL versus 65 pg/mL; P = .004), and in patients who required an intra-aortic balloon pump (IABP) (325 pg/mL versus 68 pg/mL; P= .021). Analysis of receiver operating characteristic curves demonstrated the preoperative BNP level to be a predictor of new-onset AF, a need for inotropic support, and a requirement for an IABP (areas under the curve, 0.70, 0.70, and 0.79, respectively). BNP cutoff values of 100 pg/mL for AF, 185 pg/mL for inotropic support, and 235 pg/mL for requiring an IABP predicted these postoperative adverse outcomes with 65%, 73%, and 84% accuracy, respectively. CONCLUSION: This study suggests that a higher baseline plasma BNP concentration is associated with postoperative new-onset AF, a need for inotropic support, and an IABP requirement in patients who undergo first-time isolated CABG. PMID- 18430663 TI - Retrospective analysis of surgically treated infective endocarditis cases. AB - BACKGROUND: Prosthetic valve endocarditis (PVE) and native valve endocarditis (NVE) both cause high rates of morbidity and mortality and are significant health problems in our community. Optimal timing of the surgical intervention depends on the hemodynamic stability of the patient. In the present study, we retrospectively evaluated the clinical status, bacteriology, morbidity, and mortality parameters of infective endocarditis cases that were treated surgically. METHODS: Thirty patients (20 male and 10 female) who underwent cardiac valve surgery between April 2001 and December 2006 were included in the study. The mean (SD) age of the patients was 36.5 +/- 5.42 years. Thirty-five surgical operations were conducted on 30 patients. We evaluated the patient demographic, etiologic, and surgical data retrospectively with respect to mortality and morbidity. RESULTS: The mean time to develop PVE was 13 months. We recorded a mortality rate of 16.6% (2 deaths in NVE operations and 3 deaths in PVE operations). Repeat surgeries were performed in 2 aortic valve cases and 3 mitral valve cases in which paravalvular leakage was noticed in the prosthetic valves. CONCLUSION: Despite significant medical and surgical advances, both NVE and PVE still continue to be causes of high mortality and morbidity rates in cardiac surgery. PMID- 18430664 TI - Bartonella quintana and Bartonella pediococcus Infection after Aortic Valve Replacement. AB - Bartonella quintana and Bartonella pediococcus infections are very rare causes of endocarditis. Urban trench fever with relapsing febrile illness, headache, leg pain, and endocarditis has now begun to be a more important cause of disease in socially disadvantaged persons. The diagnosis is difficult because the growth of B. quintana in blood culture takes 20-40 days. B. pediococcus may be an opportunistic pathogen in severely compromised hosts, although it has been described as a harmless bacterium. We describe a patient who developed bioprosthetic valve infection with B. quintana and B. pediococcus after valve replacement. PMID- 18430665 TI - Aortic valve replacement in a patient with morquio syndrome. AB - Cardiac involvement in Morquio syndrome, mucopolysaccharidosis IV, is characterized by aortic and mitral valve thickening and infiltration of the coronary arteries. There are few reports concerning surgical interventions in patients with mucopolysaccharidoses. We report a case of a patient affected by Morquio syndrome who underwent aortic valve replacement surgery for severe aortic valve stenosis, with an uneventful postoperative course. Cardiac surgery appears safe and feasible in these patients and improves the quality of life, even if the future prognosis related to the underlying disorder should be considered. PMID- 18430666 TI - Selective retrograde venous revascularization of the myocardium when PCI or CABG is impossible: investigation in a porcine model. AB - We investigated the possibility of nourishing the myocardium through selective retrograde coronary venous bypass grafting (CVBG) with an off-pump technique and evaluated various methods of monitoring the physiological effects of this procedure. In a porcine model, the left internal mammary artery (LIMA) was anastomosed to the left anterior descending coronary vein (LAD vein) in an off pump procedure. The LAD vein was ligated proximal to the anastomosis. The LAD artery was ligated proximally. The physiological effects were monitored using microdialysis, tissue oxygen tension, blood flow in LIMA, blood samples, and hemodynamic and histological analyses. As controls, 5 pigs underwent surgery involving only LAD artery ligation without CVBG. CVBG with LAD ligation was performed in 16 pigs; 12 survived CVBG and were monitored for 2-2.5 hours while in sinus rhythm, a 75% salvage rate after an otherwise lethal LAD artery occlusion. Immediately after LAD artery ligation, the anterior wall of the left ventricle became cyanotic and hypokinetic. Over time it regained color and contractility as flow in the LIMA increased. Microdialysis showed a significant increase in lactate. Initially tissue oxygen tension decreased, but with time some recovery was seen. Cardiac troponin T was elevated. Histological analysis showed ischemic changes. In control pigs, microdialysis was performed for 1.5 hours up to LAD artery ligation, after which all pigs died in ventricular fibrillation arrest. No increase in lactate was observed. These results indicate that after LAD artery occlusion, CVBG can nourish the myocardium to a certain extent and prevent death in the majority of cases, although varying degrees of ischemia remain. PMID- 18430667 TI - Mating system and avpr1a promoter variation in primates. AB - It has been suggested that primate mating and social behaviours may be influenced by variation in promoter region repetitive DNA of the vasopressin receptor 1a gene (avpr1a). We show that male mating behaviour does not covary in a simple way with promoter repetitive DNA in 12 Old World primates. We found that one microsatellite (-553 bp upstream) was present in all species, irrespective of their behaviour. By contrast, two microsatellites (-3956 and -3625 bp upstream) were present only in some species, yet this variation did not correlate with behaviour. These findings agree with a recent comparative analysis of voles and show that the variation in repetitive DNA in the avpr1a promoter region does not generally explain variation in male mating behaviour. Phylogenetic analysis revealed a GAGTA motif that has been independently deleted three times and involved in another larger deletion. Importantly, the presence/absence of this GAGTA motif leads to changes in predicted transcription factor-binding sites. Given the repeated loss of this motif, we speculate that it might be of functional relevance. We suggest that such non-repetitive variation, either in indels or in sequence variation, are likely to be important in explaining interspecific variation in avpr1a expression. PMID- 18430668 TI - A quick guide to video-tracking birds. AB - Video tracking is a powerful new tool for studying natural undisturbed behaviour in a wide range of birds, mammals and reptiles. Using integrated animal-borne video tags, video footage and positional data are recorded simultaneously from wild free-ranging animals. At the analysis stage, video scenes are linked to radio fixes, yielding an animal's eye view of resource use and social interactions along a known movement trajectory. Here, we provide a brief description of our basic equipment and field techniques to enable other researchers to start their own video-tracking studies. PMID- 18430669 TI - Stroke and design: background and context. PMID- 18430670 TI - The Chicago perspective on design for the disabled. AB - This article describes the efforts made by the city of Chicago to promote total access and full participation for people with disabilities. The process that the Chicago Mayor's Office for People with Disabilities (MOPD) undertook to ensure that other city departments and sister agencies incorporated accessible programs and physical accessibility considerations into their own buildings, programs, and processes is described. Some of the major accomplishments made by the department include the addition of an accessibility review added to the building permit process and a rewrite of the Chicago Building Code that requires more accessibility features in single-family homes and smaller apartment buildings. Examples are given of how thinking beyond the minimum accessibility requirements can better meet the needs of people with a disability. PMID- 18430671 TI - Creating engaging experiences for rehabilitation. AB - The traditional model of rehabilitation center design based on usability and function falls short of addressing the aspirations of those who use them. To better serve the motivational needs of both patients and therapists, we need to reconsider the gymnasium-inspired designs of current rehabilitation centers. Designers Patricia Moore and David Guynes have drawn inspiration from the everyday to create more engaging rehabilitation experiences with their Easy Street, Independence Square, Rehab 1-2-3, Our Town, and WorkSyms rehabilitation environments. Their designs simulate real-life situations to motivate patients by helping them connect their therapy to the life in which they aspire to return. Utilizing an empathic research process, Moore and Guynes build a deeper understanding of both patients' and therapists' values and apply that understanding to designs that are more directly connected to patients' aspirational goals while still meeting their functional rehabilitation needs. This same research-based design approach is utilized in all of their design work that has included, most recently, the design of the Phoenix Valley Transit Authority's Metro Light Rail Train. The train and stations have won awards for accessibility and will begin public operation in late 2008. PMID- 18430672 TI - Role of persons with a disability in the design process. AB - People with disabilities are profoundly affected by what is built around and for them, yet far too often they are not consulted during the various phases of a product's life cycle. People with disabilities need to be strong self-advocates and demand relevance to ensure that researchers ask the right questions and that engineers and designers develop products that will meet their needs. This article discusses the need for consumer involvement in all aspects of the design process, from initial concept to research and development and to influencing technology transfer and marketing. PMID- 18430673 TI - Accessibility for all: a case study of the access living headquarters. AB - Access Living of Metropolitan Chicago is a nonresidential Center for Independent Living for people with all types of disabilities. This article describes the process of creating an accessible environment for their new headquarters. A charrette involving members of the Chicago disability community, as well as experts on accessibility and Universal Design, was conducted to investigate how architecture and technology could be integrated into the design that would help improve the function of the building. Design solutions for the building entrance, bathrooms, lighting, work stations, and elevators are described in this article. By understanding what the client needed and by doing careful research to find the right products, conventional products were used almost 100% of the time. With the exception of the TTYs in the elevator, there were no customized products. This meant that there was no need to pay more for the Universal Design when it came to building products. PMID- 18430674 TI - The live well collaborative: a new model for universities and companies to work together to meet the needs of 50+ consumers. AB - A new opportunity has emerged for universities and corporations to align in response to the global trend of countries with significant percentages of their population over the age of 50. This trend is driven by the dramatic increase in the birth rate starting in 1945 after World War II and ending in 1965 (a population commonly referred to in the United States as "baby boomers"). As they reach retirement, this age cohort has a unique expectation of a continued high quality of life in spite of the emerging health challenges they face. The boomer's lifestyle expectation has tremendous impact on the design of new products and services in every sector of the economy. To achieve effective translational research (from laboratory to marketable products), universities and companies must integrate the qualitative innovation processes used by design fields like industrial design into the quantitative research techniques embraced by medicine and engineering. The University of Cincinnati is one of the few universities in the United States with all the necessary components in place to respond to this opportunity. P&G with global headquarters in Cincinnati is the right corporate partner to define this new university-corporate relationship. PMID- 18430675 TI - Developing more desirable products for stroke survivors. AB - This article examines the user-centered design process of developing a desirable assistive technology: an egg cracker to be used by stroke survivors. The case study explores the differences involved in the processes used to develop assistive technologies and consumer products. It demonstrates that combining elements from both processes can lead to the development of innovative product ideas that would not be uncovered through either process alone. Specifically, the article suggests that developing an assistive technology as a consumer product should result in a more appealing product for stroke survivors and potentially a more functional product for a wider variety of users. PMID- 18430676 TI - Success stories in design: the Versluis Park. AB - This project's initiative was to go above and beyond the Americans with Disability Act (ADA) to provide universal access to a waterfront township park in Plainfield Township, Michigan. The challenges, defined by topography, fluctuating lake levels, and a lake/beach environment, had to be overcome by nontraditional methods. Creative solutions were used to provide access for individuals with a disability to engage Versluis Lake. A unique water access ramp and transfer station provide opportunities to enter the water. A viewing deck with companion seating areas allows friends and families to sit together side by side in a social setting. A floating pier with accessible fishing stations and seating creates another recreation opportunity. Nontraditional, creative designs can empower people with disabilities to recreate along a park's waterfront. PMID- 18430677 TI - Designing the experience of health care. AB - New ideas and information services will be best served when they are examined from both the users' and the corporate perspective. It is imperative to identify what best serves the needs of the user while understanding the corporate context that would allow these products/services to be sustainable. The desired result is for users to feel included and become more active and involved in their lives and to be able to do so at a reasonable, sustainable price point. This collaboration is illustrated by several examples of health care product development including the development of a medication management system for patients and a diet assistance program for cardiac patients. In these examples, success is dependent on a solid relationship between people who have the methods to understand users and to develop products and people who have knowledge of the health care field and understand it in a business context. PMID- 18430678 TI - KineAssist: design and development of a robotic overground gait and balance therapy device. AB - BACKGROUND AND PURPOSE: Balance and mobility training consists of activities that carry a high risk for falling. The purpose of this article is to describe a novel robotic system for allowing challenging, yet safe, balance and mobility training in persons at high risk for falls. METHOD: With no initial preconceptions of what device we would build, a user-needs analysis led us to focus on increasing the level of challenge to a patient's ability to maintain balance during gait training and also on maintaining direct involvement of a physical therapist (rather than attempting robotic replacement). The KineAssist is a robotic device for gait and balance training that has emerged from a unique design process of a start-up product of a small company and a team of therapists, engineers, mechanical design experts, and rehabilitation scientists. RESULTS: The KineAssist provides partial body weight support and postural control on the torso; allows many axes of motion of the trunk and pelvis; leaves the patient's legs accessible to a physical therapist's manipulation during walking; follows a patient's walking motions overground in forward, rotation, and sidestepping directions; and catches an individual who loses balance and begins to fall. DISCUSSION AND CONCLUSION: Design and development of the KineAssist proceeded more rapidly in the context of a small company than would have been possible in most institutional research contexts. A prototype KineAssist has been constructed and has received US Food and Drug Administration (FDA) classification and institutional review board clearance for initial human studies. The acceptance of KineAssist will ultimately depend on improved patient outcomes, the use of this new tool by therapists, the ease of use of the system, and the recognition of the unique value it brings to therapeutic recovery. PMID- 18430679 TI - Reflections of a family member. PMID- 18430680 TI - Executive summary. PMID- 18430681 TI - An innovative design program for engineering students. PMID- 18430682 TI - The Knee Nook. AB - Although the effects of a stroke vary, survivors often have limited use of one side of their body. Stroke survivors may wear an ankle-foot orthosis (AFO) to prevent their weak foot from dragging and hindering ambulation. Because of the added bulk of an AFO, donning a shoe becomes difficult. The design team, composed of freshman engineering students in the Engineering Design and Communication course at Northwestern University, interviewed stroke survivors to understand the problem and then constructed several prototypes as possible solutions. After stroke survivors tested and critiqued each prototype, the Knee Nook emerged as the most promising. Stroke survivors often place their weak foot on top of their strong knee, similar to the position of crossing one's legs, to allow them to easily reach their foot. Keeping the weak leg in place on the strong thigh while donning the AFO is often difficult. The Knee Nook is a hands-free device that holds the user's leg in this position. The device is placed on top of the user's strong knee and employs a neoprene pad to easily hold the weak leg over the strong knee. This design allows stroke survivors to independently don an AFO and shoe. PMID- 18430684 TI - Development of a one-handed nail clipper for stroke survivors. AB - Due to the physical effects of stroke, many stroke survivors experience difficulty clipping their nails. Much of this difficulty is due to weakness in the upper extremities, loss of dexterity, spasticity, and vision problems. Our goal was to design a device that would help stroke survivors shorten their finger nails, providing increased independence, convenience, comfort, and safety compared to their current method of cutting nails. From user interviews and observations, we developed several prototypes to solve this problem. The final design, the Step 'n Snip, consists of a finger nail clipper mounted on an inclined, rotatable metal rod. The clipper is attached to a foot pedal with a long steel bike brake cable, which the pedal then uses to actuate the clipper. The pedal is mounted on a base that is shaped such that it provides a nesting space for the clipper frame during storage; the two parts are kept together by a swiveling clip. The clipper itself is at a convenient height for users and is fitted with a nail clippings collection system. PMID- 18430683 TI - Design of a cafeteria tray for use by stroke survivors. AB - Survivors of a stroke present varying types and degrees of neurological impairments and functional deficits. They often have difficulty using one side of their body and may require assistive devices such as canes and walkers. As a result, carrying a standard cafeteria tray can pose a challenge to these individuals. Our goal was to design a cafeteria tray that could be easily used with one hand. User interviews of stroke survivors offered client needs that guided the entire design process. It was important to the stroke survivors that the device required intuitive understanding of use, effectively transported food using only one hand, supported the weight of a meal, and had a pleasing design to promote discreteness by avoiding a "disabled" appearance. PMID- 18430685 TI - Cortical stimulation for upper limb recovery following ischemic stroke: a small phase II pilot study of a fully implanted stimulator. AB - OBJECTIVE: To evaluate the feasibility of a fully implanted cortical stimulator for improving hand and arm function in patients following ischemic stroke. METHOD: Twenty-four chronic stroke patients with hemiplegia were randomized to targeted implanted cortical electrical stimulation of the motor cortex with upper limb rehabilitation therapy or rehabilitation therapy alone. RESULTS: Using repeated measures regression models, we estimated and compared treatment effects between groups over the study follow-up period. The investigational group had significantly greater mean improvements in Upper Extremity Fugl-Meyer (UEFM) scores during the 6-month follow-up period (weeks 1-24 following therapy), as compared to the control group (difference in estimated means = 3.8, p = .042). Box and Block (B & B) test improvement from baseline scores were also significantly better in the investigational group across the 6-month follow-up assessments (difference in estimated means = 3.8, p = .046). There was one report of seizure after device implant but prior to cortical stimulation and rehabilitation therapy, but no reports of neurologic decline. There were no improvements seen in the other measures assessed. CONCLUSION: Evidence suggests that cortical stimulation with rehabilitation therapy produces a lasting treatment effect in upper extremity motor control and is not associated with serious neurological complications. A larger multicenter study is underway. PMID- 18430686 TI - New adaptations after a brain attack. PMID- 18430687 TI - Defying boundaries: globalization, bureaucracy and academic exchange. PMID- 18430688 TI - SAGE publications moves Promotion & Education into the 21st century. PMID- 18430689 TI - The nutrition transition in Jordan: the political, economic and food consumption contexts. AB - This paper describes and analyzes the nutrition transition in Jordan. The nutrition transition is shifting from undernutrition of a population to a diet high in fat, sugar, and refined carbohydrates (overnutrition) and corresponding premature morbidity and mortality from ensuing chronic diseases. The paper reviews and examines secondary data on demographic, epidemiological, economic, social, and dietary changes in the region. Jordan is moving through the nutrition transition at a rapid pace. Demographic, epidemiological, economic, and social changes have been accompanied by changes in diet, food expenditure, and general health. Although mortality rates have dropped significantly, fertility rates still remain high at 3.6 children per woman, especially compared to other countries in the region. Urbanization has increased rapidly from 46% of the population residing in urban areas in 1965 to 75% currently. Declines in real per capita income have created a state in which 27% of families live under the poverty line. Dietary changes in the region, due in part to changes in food availability over the last 40 years, include a general rise in caloric intake (from 2,165 Kcals in 1965 to 3,161 Kcals in 1997) with an increasing percentage of energy supply coming from fats and cereals. Undernutrition and chronic rates of malnutrition are low compared to other developing countries; however, various sources reveal that overweight and obesity are on the rise. Obesity in Jordan tends to be higher among women (16%) than among men (10%), with a much higher prevalence of obesity among uneducated women (50%). In conclusion, overweight and obesity are an emerging problem in Jordan, especially for women. There is a high prevalence of risk factors associated with diet-related non-communicable diseases among both men and women. PMID- 18430690 TI - Health education's role in framing pornography as a public health issue: local and national strategies with international implications. AB - Pornography is a public health issue. However, since the US Surgeon General's workshop on pornography and public health reached a consensus statement about the impacts of pornography in 1986, few policy actions have been taken to deal with this public health problem, and intense discussion surrounding the regulation of pornography has continued. This debate spans a continuum between individual rights on one extreme and complete restriction of such material for society's good on the other extreme. However, during this same period much research has been done on the impacts of pornography on children and adults. This paper focuses on reviewing the effects of pornography on society including women, children, and consumers, and includes discussion on the current and failed policies aimed at regulating pornography. The growing phenomenon of Internet pornography use is discussed in depth, and specific policy ideas pertaining to Internet pornography are presented from a public health perspective. PMID- 18430691 TI - How have Global Health Initiatives impacted on health equity? AB - This review examines the impact of Global Health Initiatives (GHIs) on health equity, focusing on low- and middle-income countries. It is a summary of a literature review commissioned by the WHO Commission on the Social Determinants of Health. GHIs have emerged during the past decade as a mechanism in development assistance for health. The review focuses on three GHIs, the US President's Emergency Plan For AIDS Relief (PEPFAR), the World Bank's Multi-country AIDS Programme (MAP) and the Global Fund to Fight AIDS, TB and Malaria. All three have leveraged significant amounts of funding for their focal diseases - together these three GHIs provide an estimated two-thirds of external resources going to HIV/AIDS. This paper examines their impact on gender equity. An analysis of these Initiatives finds that they have a significant impact on health equity, including gender equity, through their processes of programme formulation and implementation, and through the activities they fund and implement, including through their impact on health systems and human resources. However, GHIs have so far paid insufficient attention to health inequities. While increasingly acknowledging equity, including gender equity, as a concern, Initiatives have so far failed to adequately translate this into programmes that address drivers of health inequity, including gender inequities. The review highlights the comparative advantage of individual GHIs, which point to an increased need for, and continued difficulties in, harmonisation of activities at country level. On the basis of this comparative analysis, key recommendations are made. They include a call for equity-sensitive targets, the collection of gender disaggregated data, the use of policy-making processes for empowerment, programmes that explicitly address causes of health inequity and impact assessments of interventions' effect on social inequities. PMID- 18430692 TI - Strategies for implementing Health-Promoting Schools in a province in China. AB - After successful pilot projects in 10 schools (four schools with tobacco control and six schools with nutrition interventions, plus 10 control schools), Health and Education officials in Zhejiang Province, China, decided to scale up Health Promoting Schools (HPS) systematically over the entire province, starting with an initial cohort of 51 additional schools, reaching from primary to vocational schools. Interviews with school personnel during the first phase of scaling up illuminated the key pre-implementation, implementation, and monitoring and evaluation activities. Pre-implementation activities included choosing an entry point, setting up a special HPS committee, and establishing a work plan. Implementation activities included conducting mobilization meetings, prioritizing health, popularizing the HPS concept, ensuring community cooperation and participation, acting as role models, offering training, and using new teaching and learning methods. Monitoring and evaluation activities included process, baseline, and final evaluations and changing standards of evaluation to a more holistic evaluation that schools go through to become Health-Promoting Schools. Schools also reported that they faced - and overcame - a number of challenges including understanding and integrating the HPS concept and lack of professional development and support. Results revealed that schools transitioned from a passive model of education to interactive pedagogy put priority on health and viewed it as a co-responsibility, reshaped assessment to a more holistic approach and called for more training and technical support. Participants mentioned that they gained knowledge and skills and developed a deeper understanding about health. Health impact was also demonstrated, for instance in reduced injuries and reduced smoking, and educational impact was demonstrated, for instance in improved relationships of children to parents and teachers, improved social qualities, and improved teacher satisfaction. PMID- 18430693 TI - Higher education in health promotion in Europe: a comparative analysis of master's level training programmes using HP-Source.net. AB - This report summarises opportunities in Europe for master's degree level training in health promotion. Using data available at www.HP-Source.net, 105 study programmes at 71 institutions, spread over 20 European countries, were identified that include health promotion as a main subject. The programmes were analysed along a number of dimensions, including title, learning objectives, curricula, learning and teaching methods, entry requirements, duration, accreditation, language(s) of instruction and participation in European educational structures. The present analysis reveals great diversity along all these dimensions, but also several clusters of programmes that offer quite similar education in health promotion. Of special interest is the range of options available in Europe for length of study, ranging from one to two years, with part-time as well as full time options. PMID- 18430696 TI - ['One risk is ok. Are three a sure damage?" Health education's obstacles in the prevention of multiple risks]. AB - Health education can have unexpected effects, sometimes even harmful, in particular when dealing with, on the one hand, multiple risk pathologies and, on the other, one risk factor that is connected to various different pathologies. Certain unexpected effects can be the result of too much information on prevention or when prevention is too focused on one target. In the first scenario, relativity of the information could arise faced with certain health risks or be used against public health objectives. When information is too focused on one target group, it can create discriminatory reactions for the groups at risk, which can translate into denial of the potential risk by those outside of the target group. Other prevention actions can have an unexpected effect, or none at all, because the target behavior is considered functional for the individual. In order to prevent a risk behavior, it is necessary to take into account the threats it is meant to avoid (concurrent risks) and other risk factors meant to attain the same objectives (substitutable risks). In order to understand and avoid different obstacles, people's beliefs and knowledge, which are often rooted in daily behavior and hence difficult to change, must be taken into account. It is also important to situate behavior in context, identifying its function and the concurrent and substitutable risks, which means researching as well the social sciences. In the future, the need for a global health approach will be evident because prevention of risk behaviors increasingly refers to multiple risks in one same pathology as well as concomitant prevention of multiple pathologies. PMID- 18430700 TI - A genuinely developmental theory of sexual enjoyment and its implications for psychoanalytic technique. AB - A small computer-assisted word frequency analysis, indicating the extent of explicit concern with sexuality in the psychoanalytic literature, has revealed an apparent decline of psychoanalytic interest in psychosexuality. The apparent decline may be related to the limitations of drive theory and object relations approaches in offering persuasive and comprehensive accounts of the psychosexual. A new model of human sexual experience is proposed, rooted in an integration of French psychoanalytic ideas with recent developmental observational research, that once again places sexuality at the center of psychoanalytic clinical inquiry. Because emotion regulation arises out of the mirroring of affect by a primary caregiver and sexual feelings are unique in that they are systematically ignored and left unmirrored by caregivers, sexual feelings remain fundamentally dysregulated in all of us. Adult sexual experience serves as a way of coming to organize the psychosexual. The model accounts for some aspects of the phenomenology of sexual arousal and suggests ways of understanding pathological distortions of sexual behavior. The nature of the psychosexual is explored in the analytic treatment of an adolescent boy. PMID- 18430701 TI - Six essays on sexuality: an introduction. PMID- 18430702 TI - The otherness of sexuality: excess. AB - The present essay, the second of a series of three, aims at developing an experience-near account of sexuality by rehabilitating the idea of excess and its place in sexual experience. It is suggested that various types of excess, such as excess of excitation (Freud), the excess of the other (Laplanche), excess beyond symbolization and the excess of the forbidden object of desire (Leviticus; Lacan) work synergistically to constitute the compelling power of sexuality. In addition to these notions, further notions of excess touch on its transformative potential. Such notions address excess that shatters psychic structures and that is actively sought so as to enable new ones to evolve (Bersani). Work is quoted that regards excess as a way of dealing with our lonely, discontinuous being by using the "excessive" cosmic energy circulating through us to achieve continuity against death (Bataille). Two contemporary analytic thinkers are engaged who deal with the object-relational and intersubjective vicissitudes of excess. PMID- 18430703 TI - Sex and shame: the inhibition of female desires. AB - A developmental narrative is presented that centers on bodily based narcissistic injury and sense of shame in response to unrequited oedipal longings. Through an experience of oedipal defeat in relation to both mother and father, a female sense of inadequacy and shame may be internalized and accepted as one's identity, in contrast to the male phallic-omnipotent trajectory. The demise of genital narcissism in females can underlie various expressions of pervasive inhibition and failure to actualize desire. The thesis offered goes beyond separation individuation theory in suggesting that girls may inhibit sexuality and aggression, and themselves more generally, due to a representation of self as "not having what it takes" genitally, and then bodily and psychically. Mental representations of the self, based on positive imagery of the female body, are needed to give voice to a woman's bodily experience and sexual desire and agency in various realms. Two clinical vignettes illustrate female inhibitions in sexuality and in professional ambition as understood within the framework presented. PMID- 18430704 TI - Women showing off: notes on female exhibitionism. AB - The limitations of the phallocentric cast of earlier psychoanalytic formulations of "female exhibitionism" linger into the present. In part this connects to certain historical expectations for women's social behavior, and to the vicissitudes of Freud's insufficient knowledge of women in his libidinal psychosexual phasing used as a basis for analytic understanding. The contemporary fade of libido theory contributes to the neglect of such topics as they relate to the biological body. Yet ease and conflict regarding conscious and unconscious female body image representations related to that stepchild of theory-pregnancy and childbirth in particular-play a major role in female body display. Recognition of such body fantasies and female body meanings from early childhood into maturity tends to be marginalized within all of the psychoanalytic theories current today. The focus here on female exhibitionism suggests a normative spectrum for pleasurably active sex seeking and pleasurable procreative desire and fantasy that is present in a female's use of her body and which (of course, but secondarily) can become caught up in conflict. Two cases accenting analyses of female "showing off" behavior are included. PMID- 18430705 TI - Masturbation, sexuality, and adaptation: normalization in adolescence. AB - During adolescence the central masturbation fantasy that is formulated during childhood takes its final form and paradoxically must now be directed outward for appropriate object finding and pair matching in the service of procreative aims. This is a step in adaptation that requires a further developmental landmark that I have called normalization. The path toward airing these private fantasies is facilitated by chumship relationships as a step toward further exposure to the social surround. Hartmann's structuring application of adaptation within psychoanalysis is used as a framework for understanding the process that simultaneously serves intrapsychic and social demands and permits goals that follow evolutionary principles. Variations in the normalization process from masturbatory isolation to a variety of forms of sexual socialization are examined in sociological data concerning current adolescent sexual behavior and in case examples that indicate some routes to normalized experience and practice. PMID- 18430706 TI - Sexual differentiation of behavior: the foundation of a developmental model of psychosexuality. AB - The sexual differentiation of the brain and behavior occurs as the result of prenatal hormonal influences. Knowledge of this area is helpful for the construction of an appropriately modern psychoanalytically informed developmental paradigm of psychosexuality. PMID- 18430707 TI - Primary mental expression: freud, klein, and beyond. AB - Freud formulated the primary process model, describing mental activity that creates the illusion of an actual experience in lieu of reflective thought, at the very start of his career. In this, his initial formulation of unconscious mental activity, he was attempting to account for the nature of dreaming, by inference for the mind of infancy and, more speculatively, for adult psychosis. He never revised the model in light of his later formulations of the structural model and the death instinct, nor did he elaborate on his speculation that it could serve as a model for psychosis, and there has been little subsequent effort to employ the model outside the context of dreaming. A small number of analysts, including Klein, Bion, and Matte-Blanco, have constructed theories of psychosis in idiosyncratic conceptual languages that seem to be describing phenomena similar to those from which Freud constructed his model. Although Klein's model of positions, which has become the most widely accepted theory of psychosis, is generally considered a fundamental departure from Freud, both accounts have remarkable similarity and both tend to confuse primary mental expression with mature thought and normal infancy with psychosis. Contributions by cognitive developmental psychologists including Werner and Piaget suggest ways to clarify some of the confusion and to supplement and amplify Freud's and Klein's description of some of the salient features of primary mental expression. Findings from neuroimaging studies of dreaming and of schizophrenia support the proposition that primary mental activity is a qualitatively distinctive form of mental expression. PMID- 18430708 TI - Toward a general theory of unconscious processes in psychoanalysis and anesthesiology. AB - Psychoanalysis and anesthesiology appear radically different in their clinical practice, yet they share a focus of inquiry: unconscious processes. Despite this common domain, there has been no exploration of the relationship between "the unconscious" as conceived by psychoanalysts and "surgical unconsciousness" as conceived by anesthesiologists. This is likely due to the fact that general anesthesia has been assumed to be a state in which the brain is simply "turned off." More recent neuroscientific data invalidate this assumption by demonstrating that the anesthetized brain is both cognitively dynamic and capable of implicit learning. Current perspectives on anesthetic mechanisms suggest that general anesthesia is characterized not simply by the absence of cognitive activity, but by the disintegration of cognitive activity. The cognitive unbinding paradigm of general anesthesia is discussed and its application to Wilfred Bion's theory of thinking, as well as his concept of attacks on linking, is elucidated. Based on the common structure and function of unconscious processes in psychoanalysis and anesthesiology, the foundation of a general theory is established. PMID- 18430709 TI - Affect integration in dreams and dreaming. AB - The processes by which dreaming aids in the ongoing integration of affects into the mind are approached here from complementary psychoanalytic and nonpsychoanalytic perspectives. One relevant notion is that the dream provides a psychological space wherein overwhelming, contradictory, or highly complex affects that under waking conditions are subject to dissociation, splitting, or disavowal may be brought together for observation by the dreaming ego. This process serves the need for psychological balance and equilibrium. A brief discussion of how the mind processes information during dreaming is followed by a consideration of four component aspects of the integrative process: the nature and use of the dream-space, the oscillating "me / not me" quality of the dream, the apparent reality of the dream, and the use of nonpathological projective identification in dreaming. Three clinical illustrations are offered and discussed. PMID- 18430710 TI - Pluralism in action. Panel report. PMID- 18430711 TI - Current views of the oedipal complex. Panel report. PMID- 18430712 TI - What use is consciousness? A clinical neuroscience roundtable. Panel report. PMID- 18430715 TI - DNA instability in low-risk myelodysplastic syndromes: refractory anemia with or without ring sideroblasts. AB - We tested genomic instability in patients with myelodysplastic syndrome (MDS) by the comet assay and verified the suitability of this approach as a tool for analysis of ineffective hematopoiesis in refractory anemia (RA) and RA with ring sideroblasts (RARS). Erythroid and myeloid cell populations from bone marrow aspirates of 20 RA, 14 RARS and 15 control subjects were separated by differential expression of glycophorin A and subjected to comet assay. The extent of DNA migration was measured in single cells (200 cells/bone marrow fraction/subject). The results were in agreement with the concept of increased apoptosis in low-risk MDS subtypes. The RA samples had a significantly higher DNA instability than controls in glycophorin A positive cells, and the extent of DNA breakage correlated with the degree of cytopenia. Although RARS had an even higher rate of genomic instability in bone marrow cells than RA, there was no clear relationship to peripheral cytopenia. This suggests an additional DNA instability of non-apoptotic origin. Whether this increase is associated with an increased repair of oxidative damage in DNA arising due to iron deposits in ring sideroblasts remains to be formally proven. Comet assay provides a promising tool for the investigation of difference between RA and RARS pathobiology. PMID- 18430717 TI - Evaluation of exposure parameters in plain radiography: a comparative study with European guidelines. AB - Typical distribution of exposure parameters in plain radiography is unknown in Portugal. This study aims to identify exposure parameters that are being used in plain radiography in the Lisbon area and to compare the collected data with European references [Commission of European Communities (CEC) guidelines]. The results show that in four examinations (skull, chest, lumbar spine and pelvis), there is a strong tendency of using exposure times above the European recommendation. The X-ray tube potential values (in kV) are below the recommended values from CEC guidelines. This study shows that at a local level (Lisbon region), radiographic practice does not comply with CEC guidelines concerning exposure techniques. Further national/local studies are recommended with the objective to improve exposure optimisation and technical procedures in plain radiography. This study also suggests the need to establish national/local diagnostic reference levels and to proceed to effective measurements for exposure optimisation. PMID- 18430716 TI - Major basic protein-1 promotes fibrosis of dystrophic muscle and attenuates the cellular immune response in muscular dystrophy. AB - The immune response to dystrophin-deficient muscle promotes the pathology of Duchenne muscular dystrophy (DMD) and the mdx mouse model of DMD. In this investigation, we find that the release of major basic protein (MBP) by eosinophils is a prominent feature of DMD and mdx dystrophy and that eosinophils lyse muscle cells in vitro by the release of MBP-1. We also show that eosinophil depletions of mdx mice by injections of anti-chemokine receptor-3 reduce muscle cell lysis, although lysis of mdx muscle membranes is not reduced by null mutation of MBP-1 in vivo. However, ablation of MBP-1 expression in mdx mice produces other effects on muscular dystrophy. First, fibrosis of muscle and hearts, a major cause of mortality in DMD, is greatly reduced by null mutation of MBP-1 in mdx mice. Furthermore, either ablation of MBP-1 or eosinophil depletion causes large increases in cytotoxic T-lymphocytes (CTLs) in mdx muscles. The increase in CTLs in MBP-1-null mice does not reflect a general shift toward a Th1 inflammatory response, because the mutation had no significant effect on the expression of interferon-gamma, inducible nitric oxide synthase or tumor necrosis factor. Rather, MBP-1 reduces the activation and proliferation of splenocytes in vitro, indicating that MBP-1 acts in a more specific immunomodulatory role to affect the inflammatory response in muscular dystrophy. Together, these findings show that eosinophil-derived MBP-1 plays a significant role in regulating muscular dystrophy by attenuating the cellular immune response and promoting tissue fibrosis that can eventually contribute to increased mortality. PMID- 18430718 TI - Results of a European dose survey for mammography. AB - For the dose study, a semi-automated method of data collection is used in this study. The participating centres were asked to fill out a spreadsheet with all necessary data and return it. For direct digital (DR) systems, the relevant data available in the DICOM header were used. All data is automatically added to a database and processed. The data were used to calculate the mean glandular dose for every image and for different thicknesses of polymethyl methacrylate phantoms using available conversion factors. Second-degree polynomials were fitted to the patient dose data and a reference dose curve was constructed for a range of thicknesses instead of a dose reference level at a single point. The dose reference curve rises from 1.57 mGy for a thickness of 30 mm to 2.50 mGy for 55 mm and 3.83 mGy for 75 mm. The results show centres that exceed this curve lie only in the lower or higher range of thicknesses and would remain undetected using a dose reference value in a single point. This gives better information to radiographers on where there is room for improvement of the dose levels in their system. PMID- 18430719 TI - Evaluation of exposure index (IgM) in orthopaedic radiography. AB - The exposure index (lgM) obtained from a radiographic image may be a useful feedback indicator to the radiographer about the appropriate exposure level in routine clinical practice. This study aims to evaluate lgM in orthopaedic radiography performed in the standard clinical environment. We analysed the lgM of 267 exposures performed with an AGFA CR system. The mean value of lgM in our sample is 2.14. A significant difference (P = 0.000 < or =0.05) from 1.96 lgM reference is shown. Data show that 72% of exposures are above the 1.96 lgM and 42% are above the limit of 2.26. Median values of lgM are above 1.96 and below 2.26 for Speed class (SC) 200 (2.16) and SC400 (2.13). The interquartile range is lower in SC400 than in SC200. Data seem to indicate that lgM values are above the manufacturer's reference of 1.96. Departmental exposure charts should be optimised to reduce the dose given to patients. PMID- 18430720 TI - Plant phenolic compounds as ethidium bromide efflux inhibitors in Mycobacterium smegmatis. AB - BACKGROUND: One-third of the world's population is infected with the dormant tuberculosis bacillus, and there have been no new antimycobacterial compounds with new modes of action for over 30 years. Extensively drug-resistant tuberculosis is resistant to first- and second-line drugs, which can have severe side effects, and requires the breakthrough of new antituberculotics and resistance-modifying agents. Efflux pumps can cause multidrug resistance and have recently evoked much interest as promising new targets in antimicrobial therapy. OBJECTIVES: The study was performed to set up an ethidium bromide (EtBr) efflux assay in Mycobacterium smegmatis mc(2)155 for testing plant natural compounds as mycobacterial efflux pump inhibitors (EPIs). METHODS: After determining the MICs of the putative EPIs, they were tested for synergistic effects with EtBr prior to the efflux assay. RESULTS: We established an EtBr efflux assay in M. smegmatis mc(2)155. The isoflavone biochanin A exhibited efflux pump inhibiting activity comparable to that of verapamil. The flavone luteolin and the stilbene resveratrol were less active. CONCLUSIONS: A new assay was established to observe the EtBr efflux in M. smegmatis and was applied to evaluate plant phenolic compounds. Our results highlighted that the isoflavonoid biochanin A exhibited better EPI activities than other flavonoids in mycobacteria. PMID- 18430721 TI - Crystal structure of HAb18G/CD147: implications for immunoglobulin superfamily homophilic adhesion. AB - CD147, a member of the immunoglobulin superfamily (IgSF), plays fundamental roles in intercellular interactions in numerous pathological and physiological processes. Importantly, our previous studies have demonstrated that HAb18G/CD147 is a novel hepatocellular carcinoma (HCC)-associated antigen, and HAb18G/CD147 stimulates adjacent fibroblasts and HCC cells to produce elevated levels of several matrix metalloproteinases, facilitating invasion and metastasis of HCC cells. In addition, HAb18G/CD147 has also been shown to be a novel universal cancer biomarker for diagnosis and prognostic assessment of a wide range of cancers. However, the structural basis underlying the multifunctional character of CD147 remains unresolved. We report here the crystal structure of the extracellular portion of HAb18G/CD147 at 2.8A resolution. The structure comprises an N-terminal IgC2 domain and a C-terminal IgI domain, which are connected by a 5 residue flexible linker. This unique C2-I domain organization is distinct from those of other IgSF members. Four homophilic dimers exist in the crystal and adopt C2-C2 and C2-I dimerization rather than V-V dimerization commonly found in other IgSF members. This type of homophilic association thus presents a novel model for homophilic interaction between C2 domains of IgSF members. Moreover, the crystal structure of HAb18G/CD147 provides a good structural explanation for the established multifunction of CD147 mediated by homo/hetero-oligomerizations and should represent a general architecture of other CD147 family members. PMID- 18430722 TI - PDK1 regulates cell proliferation and cell cycle progression through control of cyclin D1 and p27Kip1 expression. AB - PDK1 (3-phosphoinositide-dependent protein kinase 1) is a key mediator of signaling by phosphoinositide 3-kinase. To gain insight into the physiological importance of PDK1 in cell proliferation and cell cycle control, we established immortalized mouse embryonic fibroblasts (MEFs) from mice homozygous for a "floxed" allele of Pdk1 and from wild-type mice. Introduction of Cre recombinase by retrovirus-mediated gene transfer resulted in the depletion of PDK1 in Pdk1(lox/lox) MEFs but not in Pdk1(+/+) MEFs. The insulin-like growth factor-1 induced phosphorylation of various downstream effectors of PDK1, including Akt, glycogen synthase kinase 3, ribosomal protein S6, and p70 S6 kinase, was markedly inhibited in the PDK1-depleted (Pdk1-KO) MEFs. The rate of serum-induced cell proliferation was reduced; progression of the cell cycle from the G(0)-G(1) phase to the S phase was delayed, and cell cycle progression at G(2)-M phase was impaired in Pdk1-KO MEFs. These cells also manifested an increased level of p27(Kip1) expression and a reduced level of cyclin D1 expression during cell cycle progression. The defect in cell cycle progression from the G(0)-G(1) to the S phase in Pdk1-KO MEFs was rescued by forced expression of cyclin D1, whereas rescue of the defect in G(2)-M progression in these cells required both overexpression of cyclin D1 and depletion of p27(Kip1) by RNA interference. These data indicate that PDK1 plays an important role in cell proliferation and cell cycle progression by controlling the expression of both cyclin D1 and p27(Kip1). PMID- 18430723 TI - Systematic deletion of the adenovirus-associated RNAI terminal stem reveals a surprisingly active RNA inhibitor of double-stranded RNA-activated protein kinase. AB - Adenoviruses use the short noncoding RNA transcript virus-associated (VA) RNA(I) to counteract two critical elements of the host cell defense system, innate cellular immunity and RNA interference, mediated by the double-stranded RNA activated protein kinase (PKR) and Dicer/RNA-induced silencing complex, respectively. We progressively shortened the VA RNA(I) terminal stem to examine its necessity for inhibition of PKR. Each deletion, up to 15 bp into the terminal stem, resulted in a cumulative decrease in PKR inhibitory activity. Remarkably, however, despite significant apparent destabilization of the RNA structure, the final RNA mutant that lacked the entire terminal stem (TSDelta21 RNA) efficiently bound PKR and exhibited wild-type inhibitory activity. TSDelta21 RNA stability was strongly influenced by solution pH, indicating the involvement of a protonated base within the VA RNA(I) central domain tertiary structure. Gel filtration chromatography and isothermal titration calorimetry analysis indicated that wild-type VA RNA(I) and TSDelta21 RNA form similar 1:1 complexes with PKR but that the latter lacks secondary binding site(s) that might be provided by the terminal stem. Although TSDelta21 RNA bound PKR with wild-type K(d), and overall change in free energy (DeltaG), the thermodynamics of binding (DeltaH and DeltaS) were significantly altered. These results demonstrate that the VA RNA(I) terminal stem is entirely dispensable for inhibition of PKR. Potentially, VA RNA(I) is therefore a truly bi-functional RNA; Dicer processing of the VA RNA(I) terminal stem saturates the RNA interference system while generating a "mini-VA RNA(I)" molecule that remains fully active against PKR. PMID- 18430724 TI - Lipid-independent secretion of a Drosophila Wnt protein. AB - Wnt proteins comprise a large class of secreted signaling molecules with key roles during embryonic development and throughout adult life. Recently, much effort has been focused on understanding the factors that regulate Wnt signal production. For example, Porcupine and Wntless/Evi/Sprinter have been identified as being required in Wnt-producing cells for the processing and secretion of many Wnt proteins. Interestingly, in this study we find that WntD, a recently characterized Drosophila Wnt family member, does not require Porcupine or Wntless/Evi/Sprinter for its secretion or signaling activity. Because Porcupine is involved in post-translational lipid modification of Wnt proteins, we used a novel labeling method and mass spectrometry to ask whether WntD undergoes lipid modification and found that it does not. Although lipid modification is also hypothesized to be required for Wnt secretion, we find that WntD is secreted very efficiently. WntD secretion does, however, maintain a requirement for the secretory pathway component Rab1. Our results show that not all Wnt family members require lipid modification, Porcupine, or Wntless/Evi/Sprinter for secretion and suggest that different modes of secretion may exist for different Wnt proteins. PMID- 18430725 TI - Structural and biochemical properties of lipid particles from the yeast Saccharomyces cerevisiae. AB - The two most prominent neutral lipids of the yeast Saccharomyces cerevisiae, triacylglycerols (TAG) and steryl esters (SE), are synthesized by the two TAG synthases Dga1p and Lro1p and the two SE synthases Are1p and Are2p. In this study, we made use of a set of triple mutants with only one of these acyltransferases active to elucidate the contribution of each single enzyme to lipid particle (LP)/droplet formation. Depending on the remaining acyltransferases, LP from triple mutants contained only TAG or SE, respectively, with specific patterns of fatty acids and sterols. Biophysical investigations, however, revealed that individual neutral lipids strongly affected the internal structure of LP. SE form several ordered shells below the surface phospholipid monolayer of LP, whereas TAG are more or less randomly packed in the center of the LP. We propose that this structural arrangement of neutral lipids in LP may be important for their physiological role especially with respect to mobilization of TAG and SE reserves. PMID- 18430726 TI - Lipid rafts determine clustering of STIM1 in endoplasmic reticulum-plasma membrane junctions and regulation of store-operated Ca2+ entry (SOCE). AB - Store depletion induces STIM1 to aggregate and relocate into clusters at ER plasma membrane junctions where it functionally interacts with and activates plasma membrane channels that mediate store-operated Ca(2+) entry (SOCE). Thus, the site of peripheral STIM1 clusters is critical for the regulation of SOCE. However, what determines the location of the STIM1 clusters in the ER-PM junctional regions, and whether these represent specific sites in the cell is not yet known. Here we report that clustering of STIM1 in the subplasma membrane region of the cell and activation of TRPC1-dependent SOCE are determined by lipid raft domains (LRD). We show that store depletion increased partitioning of TRPC1 and STIM1 into plasma membrane LRD. TRPC1 and STIM1 associated with each other within the LRD, and this association was dynamically regulated by the status of the ER Ca(2+) store. Peripheral STIM1 clustering was independent of TRPC1. However, sequestration of membrane cholesterol attenuated thapsigargin-induced clustering of STIM1 as well as SOCE in HSG and HEK293 cells. Recruitment and association of STIM1 and TRPC1 in LRD was also decreased. Additionally STIM1(D76A), which is peripherally localized and constitutively activates SOCE in unstimulated cells, displayed a relatively higher partitioning into LRD and interaction with TRPC1, as compared with STIM1. Disruption of membrane rafts decreased peripheral STIM1(D76A) puncta, its association with TRPC1 and the constitutive SOCE. Together, these data demonstrate that intact LRD determine targeting of STIM1 clusters to ER-plasma membrane junctions following store depletion. This facilitates the functional interaction of STIM1 with TRPC1 and activation of SOCE. PMID- 18430727 TI - A three-dimensional homology model of lipid-free apolipoprotein A-IV using cross linking and mass spectrometry. AB - Human apolipoprotein A-IV (apoA-IV) is a 46-kDa exchangeable plasma protein with many proposed functions. It is involved in chylomicron assembly and secretion, protection from atherosclerosis through a variety of mechanisms, and inhibition of food intake. There is little structural basis for these proposed functions due to the lack of a solved three-dimensional structure of the protein by x-ray crystallography or NMR. Based on previous studies, we hypothesized that lipid free apoA-IV exists in a helical bundle, like other apolipoprotein family members and that regions near the N and C termini may interact. Utilizing a homobifunctional lysine cross-linking agent, we identified 21 intramolecular cross-links by mass spectrometry. These cross-links were used to constrain the building of a sequence threaded homology model using the I-TASSER server. Our results indicate that lipid-free apoA-IV does indeed exist as a complex helical bundle with the N and C termini in close proximity. This first structural model of lipid-free apoA-IV should prove useful for designing studies aimed at understanding how apoA-IV interacts with lipids and possibly with unknown protein partners. PMID- 18430728 TI - Box 2 region of the oncostatin M receptor determines specificity for recruitment of Janus kinases and STAT5 activation. AB - Human and murine oncostatin M (OSM) induce their bioactivities through a heterodimeric receptor complex consisting of gp130 and the OSM receptor (OSMR), which initiates a signaling pathway involving Janus kinases (JAKs) and transcription factors of the signal transducers and activators of transcription (STAT) family. In contrast to the signal transducing receptor subunit gp130, the OSMR allows strong activation of STAT5B. The underlying molecular mechanism, however, remained unclear. Here we demonstrate that the human and murine OSM receptors use distinct mechanisms for STAT5B activation. The human receptor contains a STAT5B recruiting tyrosine motif (Tyr837/Tyr839) C-terminal to the box 1/2 region, which is absent in the mouse receptor. In contrast, the murine receptor initiates STAT5 activation directly via the receptor bound Janus kinases. Intriguingly, the murine receptor preferentially recruits JAK2, whereas the human receptor seems to have a higher affinity for JAK1. We identify a single amino acid (Phe820) in the human receptor that is responsible for this preference. Exchange by the murine counterpart (Cys815) allows recruitment of JAK2 by the human receptor and consequently activation of STAT5B independently of receptor tyrosine motifs. STAT5B interacts directly with JAK2 only in response to activation of the murine OSMR or the mutated human OSMR. Additionally, we show that OSM-induced STAT1 phosphorylation occurs independently of receptor tyrosine motifs and is mediated directly by Janus kinases, whereas the two C-terminally located tyrosine residues Tyr917/Tyr945 of the OSMR are crucial for STAT3 activation. PMID- 18430729 TI - N-type inactivation of the potassium channel KcsA by the Shaker B "ball" peptide: mapping the inactivating peptide-binding epitope. AB - The effects of the inactivating peptide from the eukaryotic Shaker BK(+) channel (the ShB peptide) on the prokaryotic KcsA channel have been studied using patch clamp methods. The data show that the peptide induces rapid, N-type inactivation in KcsA through a process that includes functional uncoupling of channel gating. We have also employed saturation transfer difference (STD) NMR methods to map the molecular interactions between the inactivating peptide and its channel target. The results indicate that binding of the ShB peptide to KcsA involves the ortho and meta protons of Tyr(8), which exhibit the strongest STD effects; the C4H in the imidazole ring of His(16); the methyl protons of Val(4), Leu(7), and Leu(10) and the side chain amine protons of one, if not both, the Lys(18) and Lys(19) residues. When a noninactivating ShB-L7E mutant is used in the studies, binding to KcsA is still observed but involves different amino acids. Thus, the strongest STD effects are now seen on the methyl protons of Val(4) and Leu(10), whereas His(16) seems similarly affected as before. Conversely, STD effects on Tyr(8) are strongly diminished, and those on Lys(18) and/or Lys(19) are abolished. Additionally, Fourier transform infrared spectroscopy of KcsA in presence of (13)C-labeled peptide derivatives suggests that the ShB peptide, but not the ShB L7E mutant, adopts a beta-hairpin structure when bound to the KcsA channel. Indeed, docking such a beta-hairpin structure into an open pore model for K(+) channels to simulate the inactivating peptide/channel complex predicts interactions well in agreement with the experimental observations. PMID- 18430731 TI - Determination of the number of active GroES subunits in the fused heptamer GroES required for interactions with GroEL. AB - A double-heptamer ring chaperonin GroEL binds denatured substrate protein, ATP, and GroES to the same heptamer ring and encapsulates substrate into the central cavity underneath GroES where productive folding occurs. GroES is a disk-shaped heptamer, and each subunit has a GroEL-binding loop. The residues of the GroEL subunit responsible for GroES binding largely overlap those involved in substrate binding, and the mechanism by which GroES can replace the substrate when GroES binds to GroEL/substrate complex remains to be clarified. To address this question, we generated single polypeptide GroES by fusing seven subunits with various combinations of active and GroEL binding-defective subunits. Functional tests of the fused GroES variants indicated that four active GroES subunits were required for efficient formation of the stable GroEL/GroES complex and five subunits were required for the productive GroEL/substrate/GroES complex. An increase in the number of defective GroES subunits resulted in a slowing of encapsulation and folding. These results indicate the presence of an intermediate GroEL/substrate/GroES complex in which the substrate and GroES bind to GroEL by sharing seven common binding sites. PMID- 18430730 TI - Hypoxia-mediated selective mRNA translation by an internal ribosome entry site independent mechanism. AB - Although it is advantageous for hypoxic cells to inhibit protein synthesis and conserve energy, it is also important to translate mRNAs critical for adaptive responses to hypoxic stress. Because internal ribosome entry sites (IRES) have been postulated to mediate this preferential synthesis, we analyzed the 5 ' untranslated regions from a panel of stress-regulated mRNAs for m(7)GTP cap independent translation and identified putative IRES elements in encephalomyocarditis virus, vascular endothelial growth factor, hypoxia-inducible factors (HIFs) 1alpha and 2alpha, glucose transporter-like protein 1, p57(Kip2), La, BiP, and triose phosphate isomerase transcripts. However, when capped and polyadenylated dicistronic RNAs were synthesized in vitro and transfected into cells, cellular IRES-mediated translation accounted for less than 1% that of the level of cap-dependent translation. Moreover, hypoxic stress failed to activate cap-independent synthesis, indicating that it is unlikely that this is the primary mechanism for the maintenance of the translation of these mRNAs under low O(2). Furthermore, although HIF-1alpha is frequently cited as an example of an mRNA that is preferentially translated, we demonstrate that under different levels and durations of hypoxic stress, changes in newly synthesized HIF-1alpha and beta-actin protein levels mirror alterations in corresponding mRNA abundance. In addition, our data suggest that cyclin-dependent kinase inhibitor p57(Kip2) and vascular endothelial growth factor mRNAs are selectively translated by an IRES-independent mechanism under hypoxic stress. PMID- 18430732 TI - Glucose-induced remodeling of intermediary and energy metabolism in procyclic Trypanosoma brucei. AB - The procyclic form of Trypanosoma brucei is a parasitic protozoan that normally dwells in the midgut of its insect vector. In vitro, this parasite prefers d glucose to l -proline as a carbon source, although this amino acid is the main carbon source available in its natural habitat. Here, we investigated how l proline is metabolized in glucose-rich and glucose-depleted conditions. Analysis of the excreted end products of (13)C-enriched l -proline metabolism showed that the amino acid is converted into succinate or l -alanine depending on the presence or absence of d-glucose, respectively. The fact that the pathway of l proline metabolism was truncated in glucose-rich conditions was confirmed by the analysis of 13 separate RNA interference-harboring or knock-out cell lines affecting different steps of this pathway. For instance, RNA interference studies revealed the loss of succinate dehydrogenase activity to be conditionally lethal only in the absence of d-glucose, confirming that in glucose-depleted conditions, l -proline needs to be converted beyond succinate. In addition, depletion of the F(0)/F(1)-ATP synthase activity by RNA interference led to cell death in glucose depleted medium, but not in glucose-rich medium. This implies that, in the presence of d-glucose, the importance of the F(0)/F(1)-ATP synthase is diminished and ATP is produced by substrate level phosphorylation. We conclude that trypanosomes develop an elaborate adaptation of their energy production pathways in response to carbon source availability. PMID- 18430733 TI - Gonadotropin-regulated testicular helicase (DDX25), an essential regulator of spermatogenesis, prevents testicular germ cell apoptosis. AB - Gonadotropin-regulated testicular helicase (GRTH)/DDX25 is an essential post transcriptional regulator of spermatogenesis. In GRTH null mice severe apoptosis was observed in spermatocytes entering the metaphase of meiosis. Pro- and anti apoptotic factors were found to be under GRTH regulation in comparative studies of spermatocytes from wild type and GRTH(-/-) knock-out (KO) mice. KO mice displayed decreased levels of Bcl-2 and Bcl-xL (anti-apoptotic factors), an increase in Bid, Bak, and Bad (pro-apoptotic), reduced phospho-Bad, and release of cytochrome c. Also, an increase on Smac, a competitor of inhibitor apoptotic proteins that release caspases, was observed. These changes caused an increase in cleavage of caspases 9 and 3, activation of caspase 3 and increases in cleavage products of PARP. The half-life of caspase 3 transcripts was markedly increased in KO, indicating that GRTH had a negative role on its mRNA stability. IkappaBalpha, which sequesters NF-kappaB from its transcriptional activation of pro-apoptotic genes, was highly elevated in KO, and its phospho-form, which promotes its dissociation, was reduced. The increase of HDAC1 and abolition of p300 expression in KO indicated a nuclear action of GRTH on the NF-kappaB mediated transcription of anti-apoptotic genes. It also regulates the associated death domain pathway and caspase 8-mediated events. GRTH-mediated apoptotic regulation was further indicated by its selective binding to pro- and anti apoptotic mRNAs. These studies have demonstrated that GRTH, as a component of mRNP particles, acts as a negative regulator of the tumor necrosis factor receptor 1 and caspase pathways and promotes NF-kappaB function to control apoptosis in spermatocytes of adult mice. PMID- 18430734 TI - N-wasp and the arp2/3 complex are critical regulators of actin in the development of dendritic spines and synapses. AB - Changes in the number, size, and shape of dendritic spines are associated with synaptic plasticity, which underlies cognitive functions such as learning and memory. This plasticity is attributed to reorganization of actin, but the molecular signals that regulate this process are poorly understood. In this study, we show neural Wiskott-Aldrich syndrome protein (N-WASP) regulates the formation of dendritic spines and synapses in hippocampal neurons. N-WASP localized to spines and active, functional synapses as shown by loading with FM4 64 dye. Knock down of endogenous N-WASP expression by RNA interference or inhibition of its activity by treatment with a specific inhibitor, wiskostatin, caused a significant decrease in the number of spines and excitatory synapses. Deletion of the C-terminal VCA region of N-WASP, which binds and activates the actin-related protein 2/3 (Arp2/3) complex, dramatically decreased the number of spines and synapses, suggesting activation of the Arp2/3 complex is critical for spine and synapse formation. Consistent with this, Arp3, like N-WASP, was enriched in spines and excitatory synapses and knock down of Arp3 expression impaired spine and synapse formation. A similar defect in spine and synapse formation was observed when expression of an N-WASP activator, Cdc42, was knocked down. Thus, activation of N-WASP and, subsequently, the Arp2/3 complex appears to be an important molecular signal for regulating spines and synapses. Arp2/3 mediated branching of actin could be a mechanism by which dendritic spine heads enlarge and subsequently mature. Collectively, our results point to a critical role for N-WASP and the Arp2/3 complex in spine and synapse formation. PMID- 18430735 TI - Enzymatic characteristics of I213T mutant presenilin-1/gamma-secretase in cell models and knock-in mouse brains: familial Alzheimer disease-linked mutation impairs gamma-site cleavage of amyloid precursor protein C-terminal fragment beta. AB - Presenilin (PS)/gamma-secretase-mediated intramembranous proteolysis of amyloid precursor protein produces amyloid beta (Abeta) peptides in which Abeta species of different lengths are generated through multiple cleavages at the gamma-, zeta , and epsilon-sites. An increased Abeta42/Abeta40 ratio is a common characteristic of most cases of familial Alzheimer disease (FAD)-linked PS mutations. However, the molecular mechanisms underlying amyloid precursor protein proteolysis leading to increased Abeta42/Abeta40 ratios still remain unclear. Here, we report our findings on the enzymatic analysis of gamma-secretase derived from I213T mutant PS1-expressing PS1/PS2-deficient (PS(-/-)) cells and from the brains of I213T mutant PS1 knock-in mice. Kinetics analyses revealed that the FAD mutation reduced de novo Abeta generation, suggesting that mutation impairs the total catalytic rate of gamma-secretase. Analysis of each Abeta species revealed that the FAD mutation specifically reduced Abeta40 levels more drastically than Abeta42 levels, leading to an increased Abeta42/Abeta40 ratio. By contrast, the FAD mutation increased the generation of longer Abeta species such as Abeta43, Abeta45, and >Abeta46. These results were confirmed by analyses of gamma secretase derived from I213T knock-in mouse brains, in which the reduction of de novo Abeta generation was mutant allele dose-dependent. Our findings clearly indicate that the mechanism underlying the increased Abeta42/Abeta40 ratio observed in cases of FAD mutations is related to the differential inhibition of gamma-site cleavage reactions, in which the reaction producing Abeta40 is subject to more inhibition than that producing Abeta42. Our results also provide novel insight into how enhancing the generation of longer Abetas may contribute to Alzheimer disease onset. PMID- 18430736 TI - Molecular features governing the stability and specificity of functional complex formation by Mycobacterium tuberculosis CFP-10/ESAT-6 family proteins. AB - The Mycobacterium tuberculosis complex CFP-10/ESAT-6 family proteins play essential but poorly defined roles in tuberculosis pathogenesis. In this article we report the results of detailed spectroscopic studies of several members of the CFP-10/ESAT-6 family. This work shows that the CFP-10/ESAT-6 related proteins, Rv0287 and Rv0288, form a tight 1:1 complex, which is predominantly helical in structure and is predicted to closely resemble the complex formed by CFP-10 and ESAT-6. In addition, the Rv0287.Rv0288 complex was found to be significantly more stable to both chemical and temperature induced denaturation than CFP-10.ESAT-6. This approach demonstrated that neither Rv0287.Rv0288 nor the CFP-10.ESAT-6 complexes are destabilized at low pH (4.5), indicating that even in low pH environments, such as the mature phagosome, both Rv0287.Rv0288 and CFP-10.ESAT-6 undoubtedly function as complexes rather than individual proteins. Analysis of the structure of the CFP-10.ESAT-6 complex and optimized amino acid sequence alignments of M. tuberculosis CFP-10/ESAT-6 family proteins revealed that residues involved in the intramolecular contacts between helices are conserved across the CFP-10/ESAT-6 family, but not those involved in primarily intermolecular contacts. This analysis identified the molecular basis for the specificity and stability of complex formation between CFP-10/ESAT-6 family proteins, and indicates that the formation of functional complexes with key roles in pathogenesis will be limited to genome partners, or very closely related family members, such as Rv0287/Rv0288 and Rv3019c/Rv3020c. PMID- 18430737 TI - T-cell development and function are modulated by dual specificity phosphatase DUSP5. AB - Interleukin-2 (IL-2) is a pleiotropic cytokine that regulates lymphocyte proliferation and peripheral tolerance. IL-2 activates mitogen-activated protein kinase (MAPK), phosphatidylinositol 3-kinase, and signal transducer and activator of transcription (STAT) pathways and modulates expression of target genes. Systematic analysis of IL-2 target genes has revealed regulation of potential feedback inhibitors of IL-2 signaling, including several suppressor of cytokine signaling (SOCS) family members as well as MAPK pathway-regulating dual specificity phosphatases (DUSPs). Here we have evaluated the in vivo actions of DUSP5, an extracellular signal-regulated kinase 1/2 (ERK1/2)-specific phosphatase, by generating transgenic mice overexpressing DUSP5 within the lymphoid compartment. We show that transgenic DUSP5 expression results in a block in thymocyte development at the double positive stage. We also demonstrate that DUSP5-expressing mature T cells exhibit decreased IL-2-dependent proliferation and defective IL-2-mediated induction of genes. Finally, DUSP5 transgenic mice develop autoimmune symptoms, suggesting a role for the MAPK pathway in the regulation of tolerance. Thus, proper regulation of DUSP5 activity is critical for normal immune system development, IL-2 actions, and tolerance. PMID- 18430738 TI - Functional consequences of the human cardiac troponin I hypertrophic cardiomyopathy mutation R145G in transgenic mice. AB - In this study, we addressed the functional consequences of the human cardiac troponin I (hcTnI) hypertrophic cardiomyopathy R145G mutation in transgenic mice. Simultaneous measurements of ATPase activity and force in skinned papillary fibers from hcTnI R145G transgenic mice (Tg-R145G) versus hcTnI wild type transgenic mice (Tg-WT) showed a significant decrease in the maximal Ca(2+) activated force without changes in the maximal ATPase activity and an increase in the Ca(2+) sensitivity of both ATPase and force development. No difference in the cross-bridge turnover rate was observed at the same level of cross-bridge attachment (activation state), showing that changes in Ca(2+) sensitivity were not due to changes in cross-bridge kinetics. Energy cost calculations demonstrated higher energy consumption in Tg-R145G fibers compared with Tg-WT fibers. The addition of 3 mm 2,3-butanedione monoxime at pCa 9.0 showed that there was approximately 2-4% of force generating cross-bridges attached in Tg R145G fibers compared with less than 1.0% in Tg-WT fibers, suggesting that the mutation impairs the ability of the cardiac troponin complex to fully inhibit cross-bridge attachment under relaxing conditions. Prolonged force and intracellular [Ca(2+)] transients in electrically stimulated intact papillary muscles were observed in Tg-R145G compared with Tg-WT. These results suggest that the phenotype of hypertrophic cardiomyopathy is most likely caused by the compensatory mechanisms in the cardiovascular system that are activated by 1) higher energy cost in the heart resulting from a significant decrease in average force per cross-bridge, 2) slowed relaxation (diastolic dysfunction) caused by prolonged [Ca(2+)] and force transients, and 3) an inability of the cardiac TnI to completely inhibit activation in the absence of Ca(2+) in Tg-R145G mice. PMID- 18430739 TI - Enhancer of zeste homologue 2 (EZH2) down-regulates RUNX3 by increasing histone H3 methylation. AB - Overexpression of enhancer of zeste homologue 2 (EZH2) occurs in various malignancies and is associated with a poor prognosis, especially because of increased cancer cell proliferation. In this study we found an inverse correlation between EZH2 and RUNX3 gene expression in five cancer cell lines, i.e. gastric, breast, prostate, colon, and pancreatic cancer cell lines. Chromatin immunoprecipitation assay showed an association between EZH2 bound to the RUNX3 gene promoter, and trimethylated histone H3 at lysine 27, and HDAC1 (histone deacetylase 1) bound to the RUNX3 gene promoter in cancer cells. RNA interference-mediated knockdown of EZH2 resulted in a decrease in H3K27 trimethylation and unbound HDAC1 and an increase in expression of the RUNX3 gene. Restoration of RUNX3 expression was not associated with any change in DNA methylation status in the RUNX3 promoter region. RUNX3 was repressed by histone deacetylation and hypermethylation of a CpG island in the promoter region and restored by trichostatin A or/and 5-aza-2'-deoxycytidine. Immunofluorescence staining confirmed restoration of expression of the RUNX3 protein after knockdown of EZH2 and its restoration resulted in decreased cell proliferation. In vivo, an inverse relationship between expression of the EZH2 and RUNX3 proteins was observed at the individual cell level in gastric cancer patients in the absence of DNA methylation in the RUNX3 promoter region. The results showed that RUNX3 is a target for repression by EZH2 and indicated an underlying mechanism of the functional role of EZH2 overexpression on cancer cell proliferation. PMID- 18430740 TI - The crystal structure of pectate lyase peli from soft rot pathogen Erwinia chrysanthemi in complex with its substrate. AB - The crystallographic structure of the family 3 polysaccharide lyase (PL-3) PelI from Erwinia chrysanthemi has been solved to 1.45 A resolution. It consists of an N-terminal domain harboring a fibronectin type III fold linked to a catalytic domain displaying a parallel beta-helix topology. The N-terminal domain is located away from the active site and is not involved in the catalytic process. After secretion in planta, the two domains are separated by E. chrysanthemi proteases. This event turns on the hypersensitive response of the host. The structure of the single catalytic domain determined to 2.1 A resolution shows that the domain separation unveils a "Velcro"-like motif of asparagines, which might be recognized by a plant receptor. The structure of PelI in complex with its substrate, a tetragalacturonate, has been solved to 2.3 A resolution. The sugar binds from subsites -2 to +2 in one monomer of the asymmetric unit, although it lies on subsites -1 to +3 in the other. These two "Michaelis complexes" have never been observed simultaneously before and are consistent with the dual mode of bond cleavage in this substrate. The bound sugar adopts a mixed 2(1) and 3(1) helical conformation similar to that reported in inactive mutants from families PL-1 and PL-10. However, our study suggests that the catalytic base in PelI is not a conventional arginine but a lysine as proposed in family PL-9. PMID- 18430741 TI - A physiologically based pharmacokinetic model for intravenous and ingested dimethylarsinic acid in mice. AB - A physiologically based pharmacokinetic (PBPK) model for the organoarsenical dimethylarsinic acid (DMA(V)) was developed in mice. The model was calibrated using tissue time course data from multiple tissues in mice administered DMA(V) intravenously. The final model structure was based on diffusion limitation kinetics. In general, PBPK models use the assumption of blood flow-limited transport into tissues. This assumption has historically worked for small lipophilic organic solvents. However, the conditions under which flow-limited kinetics occurs and how to distinguish when flow-limited versus diffusion-limited transport is more appropriate, have been rarely evaluated. One important goal of this modeling effort was to systematically evaluate descriptions of flow-limited compared with diffusion-limited tissue distribution for DMA(V), using the relatively extensive pharmacokinetic data available in mice. The diffusion limited model consistently provided an improved fit over flow-limited simulations when compared with tissue time course iv experimental data. After model calibration, an independent data set obtained by oral gavage of DMA(V), was used to further test model structure. Sensitivity analysis of the two PBPK model structures showed the importance of early time course data collection, and the impact of diffusion for kidney time course data description. In summary, this modeling effort suggests the importance of availability of organ specific time course data sets necessary for the discernment of PBPK modeling structure, motivated by knowledge of biology, and providing necessary feedback between experimental design and biological modelers. PMID- 18430742 TI - OnD-CRF: predicting order and disorder in proteins using [corrected] conditional random fields. AB - MOTIVATION: Order and Disorder prediction using Conditional Random Fields (OnD CRF) is a new method for accurately predicting the transition between structured and mobile or disordered regions in proteins. OnD-CRF applies CRFs relying on features which are generated from the amino acids sequence and from secondary structure prediction. Benchmarking results based on CASP7 targets, and evaluation with respect to several CASP criteria, rank the OnD-CRF model highest among the fully automatic server group. AVAILABILITY: http://babel.ucmp.umu.se/ond-crf/ PMID- 18430743 TI - Pre- and post-harvest influences on seed dormancy status of an Australian Goodeniaceae species, Goodenia fascicularis. AB - BACKGROUND AND AIMS: The period during which seeds develop on the parent plant has been found to affect many seed characteristics, including dormancy, through interactions with the environment. Goodenia fascicularis (Goodeniaceae) seeds were used to investigate whether seeds of an Australian native forb, harvested from different environments and produced at different stages of the reproductive period, differ in dormancy status. METHODS: During the reproductive phase, plants were grown ex situ in warm (39/21 degrees C) or cool (26/13 degrees C) conditions, with adequate or limited water availability. The physiological dormancy of resulting seeds was measured in terms of the germination response to warm stratification (34/20 degrees C, 100 % RH, darkness). KEY RESULTS: Plants in the cool environment were tall and had high above-ground biomass, yet yielded fewer seeds over a shorter, later harvest period when compared with plants in the warm environment. Seeds from the cool environment also had higher viability and greater mass, despite a significant proportion (7 % from the cool-wet environment) containing no obvious embryo. In the warm environment, the reproductive phase was accelerated and plants produced more seeds despite being shorter and having lower above-ground biomass than those in the cool environment. Ten weeks of warm stratification alleviated physiological dormancy in seeds from all treatments resulting in 80-100 % germination. Seeds that developed at warm temperatures were less dormant (i.e. germination percentages were higher) than seeds from the cool environment. Water availability had less effect on plant and seed traits than air temperature, although plants with reduced soil moisture were shorter, had lower biomass and produced fewer, less dormant seeds than plants watered regularly. CONCLUSIONS: Goodenia fascicularis seeds are likely to exhibit physiological dormancy regardless of the maternal environment. However, seeds collected from warm, dry environments are likely to be more responsive to warm stratification than seeds from cooler, wetter environments. PMID- 18430744 TI - Lornoxicam characteristically modulates cerebral pain-processing in human volunteers: a functional magnetic resonance imaging study. AB - BACKGROUND: Lornoxicam like other non-steroidal anti-inflammatory drugs (NSAIDs) is widely used for postoperative pain therapy. Evaluation of the effect of lornoxicam on cerebral processing of surgical pain was thus the aim of the present functional magnetic resonance imaging (fMRI) study. METHODS: An fMRI compatible pain model that mimics surgical pain was used to induce pain rated 4-5 on a visual analogue scale (VAS) at the anterior margin of the right tibia in volunteers (n=22) after i.v. administration of saline (n=11) or lornoxicam (0.1 mg kg(-1)) (n=11). RESULTS: Lornoxicam, which significantly reduced pain sensation [VAS: mean (sd) 4.6 (0.7) vs 1.2 (1.5)], completely suppressed pain induced activation in the SII/operculum, anterior cingulate cortex, insula, parietal (inferior), prefrontal (inferior, medial), temporal (inferior, medial/superior) lobe, cerebellum, and contralateral (e.g. left-sided) postcentral gyrus (SI). Only the hippocampus and the contralateral superior parietal lobe (BA 7) were activated. CONCLUSIONS: As compared with saline, lornoxicam typically suppressed pain-induced brain activation in all regions except the hippocampus. Furthermore, de novo activation was found in the contralateral, superior parietal lobe (BA 7). PMID- 18430745 TI - Pain in children: recent advances and ongoing challenges. AB - Significant advances in the assessment and management of acute pain in children have been made, and are supported by an increase in the availability and accessibility of evidence-based data. However, methodological and practical issues in the design and performance of clinical paediatric trials limit the quantity, and may influence the quality, of current data, which lags behind that available for adult practice. Collaborations within research networks, which incorporate both preclinical and clinical studies, may increase the feasibility and specificity of future trials. In early life, the developing nervous system responds differently to pain, analgesia, and injury, resulting in effects not seen in later life and which may have long-term consequences. Translational laboratory studies further our understanding of developmental changes in nociceptor pathway structure and function, analgesic pharmacodynamics, and the impact of different forms of injury. Chronic pain in children has a negative impact on quality of life, resulting in social and emotional consequences for both the child and the family. Despite age-related differences in many chronic pain conditions, such as neuropathic pain, management in children is often empirically based on data from studies in adults. There is a major need for further clinical research, training of health-care providers, and increased resources, to improve management and outcomes for children with chronic pain. PMID- 18430746 TI - Nociceptin and urotensin-II concentrations in critically ill patients with sepsis. AB - BACKGROUND: The systemic inflammatory response to infection (sepsis) involves widespread organ dysfunction, including changes in immune modulation, cardiovascular derangements, and neural activation. Two neuropeptide/receptor systems, nociceptin/orphanin FQ (N/OFQ) which acts at the non-classical opioid receptor NOP and urotensin-II (U-II) which acts at the urotensin receptor (UT), have been implicated in neural, immune, and cardiovascular system function. In this study, we make measurements of these peptides in critically ill patients. METHODS: Plasma samples from 21 critically ill patients with sepsis were collected over four consecutive days. Plasma N/OFQ and U-II concentrations were determined by radioimmunoassay and compared with biochemical and clinical markers of illness severity, including serum creatinine, bilirubin, platelet and white cell counts, admission APACHE II and serial SOFA scores. RESULTS: Median (inter quartile range) admission plasma N/OFQ concentrations in sepsis were higher in patients who died within 30 days (n=4) compared with survivors (n=17); 3.0 (2.5 5.0) vs 1.0 (1.0-2.5) pg ml(-1) (P=0.028). Plasma N/OFQ concentrations were increased in a subgroup of five patients who had undergone major gastrointestinal surgery. There were no significant changes in plasma U-II concentrations. There were no correlations between plasma U-II and N/OFQ concentrations and markers of illness severity and organ system dysfunction. CONCLUSIONS: Plasma N/OFQ concentrations were increased in critically ill patients with sepsis who had undergone major gastrointestinal surgery and in patients who subsequently died. Further work is required to clarify the significance of plasma N/OFQ concentrations in sepsis. PMID- 18430747 TI - Do primary care physicians treating minority patients report problems delivering high-quality care? AB - Racial and ethnic disparities in primary health care likely reflect the aggregate socioeconomic composition of a physician's patient panels as well as differences in individual patients' characteristics. National physician survey data indicate that physicians in high-minority practices depend more on low-paying Medicaid, receive lower private insurance reimbursements, and have lower incomes. These constrained resources help explain the greater quality-related difficulties delivering care reported by these physicians--such as coordination of care, ability to spend adequate time with patients during office visits, and obtaining specialty care--that relate directly to physicians' ability to function as their patients' medical home. PMID- 18430748 TI - Using nurses and office staff to report prescribing errors in primary care. AB - OBJECTIVE: To implement a prescribing-error reporting system in primary care offices and analyze the reports. DESIGN: Descriptive analysis of a voluntary prescribing-error-reporting system SETTING: Seven primary care offices in Vermont, USA. PARTICIPANTS: One hundred and three prescribers, managers, nurses and office staff. INTERVENTION: Nurses and office staff were asked to report all communications with community pharmacists regarding prescription problems. MAIN OUTCOME MEASURES: All reports were classified by severity category, setting, error mode, prescription domain and error-producing conditions. RESULTS: All practices submitted reports, although reporting decreased by 3.6 reports per month (95% CI, -2.7 to -4.4, P<0.001, by linear regression analysis). Two hundred and sixteen reports were submitted. Nearly 90% (142/165) of errors were severity Category B (errors that did not reach the patient) according to the National Coordinating Council for Medication Error Reporting and Prevention Index for Categorizing Medication Errors. Nineteen errors reached the patient without causing harm (Category C); and 4 errors caused temporary harm requiring intervention (Category E). Errors involving strength were found in 30% of reports, including 23 prescriptions written for strengths not commercially available. Antidepressants, narcotics and antihypertensives were the most frequent drug classes reported. Participants completed an exit survey with a response rate of 84.5% (87/103). Nearly 90% (77/87) of respondents were willing to continue reporting after the study ended, however none of the participants currently submit reports. CONCLUSIONS: Nurses and office staff are a valuable resource for reporting prescribing errors. However, without ongoing reminders, the reporting system is not sustainable. PMID- 18430749 TI - Enhancement of ventricular gap-junction coupling by rotigaptide. AB - AIMS: Rotigaptide is proposed to exert its anti-arrhythmic effects by improving myocardial gap-junction communication. To directly investigate the mechanisms of rotigaptide action, we treated cultured neonatal murine ventricular cardiomyocytes with clinical pharmacological doses of rotigaptide and directly determined its effects on gap-junctional currents. METHODS AND RESULTS: Neonatal murine ventricular cardiomyocytes were enzymatically isolated and cultured for 1 4 days. Primary culture cell pairs were subjected to dual whole cell patch-clamp procedures to directly measure gap-junctional currents (I(j)) and voltage (V(j)). Rotigaptide (0-350 nM) was applied overnight or acutely perfused into 35 mm culture dishes. Rotigaptide (35-100 nM) acutely and chronically increased the resting gap-junction conductance (g(j)), and normalized steady-state minimum g(j) (G(min)) by 5-20%. Higher concentrations produced a diminishing response, which mimics the observed therapeutic efficacy of the drug. The inactivation kinetics was similarly slowed in a therapeutic concentration-dependent manner without affecting the V(j) dependence of inactivation or recovery. The effects of 0-100 nM rotigaptide on ventricular g(j) during cardiac action potential propagation were accurately modelled by computer simulations which demonstrate that clinically effective concentrations of rotigaptide can partially reverse conduction slowing due to decreases in g(j) and inactivation. CONCLUSION: These results demonstrate that therapeutic concentrations of rotigaptide increase the resting gap-junction conductance and reduce the magnitude and kinetics of steady state inactivation in a concentration-dependent manner. Rotigaptide may be effective in treating re-entrant forms of cardiac arrhythmias by improving conduction and preventing the formation of re-entrant circuits in partially uncoupled myocardium. PMID- 18430750 TI - Early atherosclerosis in humans: role of diffuse intimal thickening and extracellular matrix proteoglycans. AB - This review attempts to define the early events that lead to lesions of human atherosclerosis based on careful morphological studies in human autopsy specimens. In contrast to most small laboratory animals, diffuse intimal thickening (DIT) is present in human arteries before atherosclerosis develops, particularly in the atherosclerosis-prone arteries such as coronary arteries and abdominal aorta. In the earliest stage of atherosclerosis, lipids deposit eccentrically in the deep layer of DIT to form Type I lesions. These layers are enriched in extracellular matrix (ECM) proteoglycans such as biglycan. Following lipid deposition, macrophages appear in these regions and foam cells are observed (Type II lesions). Such observations support the 'response-to-retention' hypothesis that states that a principle early event in the pathogenesis of human atherosclerosis is the trapping and retention of lipoproteins by ECM proteoglycans followed by infiltration and accumulation of macrophages. PMID- 18430751 TI - Transcriptional control of mitochondrial biogenesis: the central role of PGC 1alpha. AB - Although the concept of energy starvation in the failing heart was proposed decades ago, still very little is known about the origin of energetic failure. Recent advances in molecular biology have started to elucidate the transcriptional events governing mitochondrial biogenesis. In particular, a great step was taken with the discovery that peroxisome proliferator-activated receptor gamma co-activator (PGC-1alpha) is the master regulator of mitochondrial biogenesis. The molecular mechanisms underlying the downregulation of PGC-1alpha and the consequent decrease in mitochondrial function in heart failure are, however, still poorly understood. Indeed, the main pathways involved in mitochondrial biogenesis are thought to be up- rather than down-regulated in pathological hypertrophy and heart failure. The current review summarizes recent advances in this field and is restricted to the heart when cardiac data are available. PMID- 18430752 TI - Protein structure databases with new web services for structural biology and biomedical research. AB - The Protein Data Bank Japan (PDBj) curates, edits and distributes protein structural data as a member of the worldwide Protein Data Bank (wwPDB) and currently processes approximately 25-30% of all deposited data in the world. Structural information is enhanced by the addition of biological and biochemical functional data as well as experimental details extracted from the literature and other databases. Several applications have been developed at PDBj for structural biology and biomedical studies: (i) a Java-based molecular graphics viewer, jV; (ii) display of electron density maps for the evaluation of structure quality; (iii) an extensive database of molecular surfaces for functional sites, eF-site, as well as a search service for similar molecular surfaces, eF-seek; (iv) identification of sequence and structural neighbors; (v) a graphical user interface to all known protein folds with links to the above applications, Protein Globe. Recent examples are shown that highlight the utility of these tools in recognizing remote homologies between pairs of protein structures and in assigning putative biochemical functions to newly determined targets from structural genomics projects. PMID- 18430753 TI - Cohort Profile: the STRIP Study (Special Turku Coronary Risk Factor Intervention Project), an Infancy-onset Dietary and Life-style Intervention Trial. PMID- 18430754 TI - Pathological and molecular features of adrenocortical carcinoma: an update. AB - The pathological diagnosis of adrenocortical carcinoma (ACC), which is based on gross and microscopic criteria, is subjective. None of the features are absolutely indicative of malignancy, although their combination in a scoring system may correctly identify ACC. The Weiss system, which is currently the most popular, combines nine morphological parameters, of which three are structural ("dark" cytoplasm, diffuse architecture, necrosis), three are cytological (atypia, mitotic count, atypical mitotic figures) and three are related to invasion (of sinusoids, veins and tumour capsule). Although there are strictly defined criteria for each feature, some are straightforward and objective, while others are potentially more problematic (diffuse architecture, necrosis, sinusoidal, venous and capsular invasions). The classification of oncocytic and paediatric adrenocortical tumours is even more challenging, as not all of the above morphological parameters are predictors of malignancy in these tumour types. As an alternative to the morphological approach, a wide array of chromosomal, genetic, molecular and immunohistochemical markers have been tested in ACC to identify reliable diagnostic and prognostic factors. Genetic and epigenetic alterations of p53, IGF-2 and molecules involved in cancer cell invasive properties seem the most promising. These molecular markers may not only play a role in the biology of these tumours and have prognostic implications, but may also be used as potential targets for treatment. However, these markers are not sufficiently sensitive and specific to replace conventional morphological criteria. PMID- 18430755 TI - Testing for JAK2V617F mutation across specimen types yields concordant results. PMID- 18430756 TI - Fine needle aspiration of the thyroid: a pathologist's perspective. PMID- 18430757 TI - Prognostic value of mitotic counts and Ki-67 immunoreactivity in adult-type granulosa cell tumour of the ovary. AB - AIMS: The identification of reliable prognostic factors in patients with ovarian stage 1 adult-type granulosa cell tumour (GCT) has proved problematic. Some reports have suggested that proliferation indices may be of value, but the data are conflicting and the methods of assessment often poorly defined. In this study the mitotic activity and Ki-67 immunohistochemistry was assessed in a series of GCT using carefully specified methodology, and the findings were correlated with clinicopathological findings. METHODS: Tumour proliferation was assessed in 38 primary GCT by counting mitotic figures in 50 high-power fields (x500 magnification) with results expressed as a mean count per 2 mm(2) standardised area. The number of mitotic figures and Ki-67 immunoreactive cells per 10,000 tumour cells was also assessed using an ocular cell counting graticule. The results were correlated with tumour stage at presentation and with the development of tumour recurrence. RESULTS: Twenty-nine patients were stage 1 at presentation, and nine patients had high-stage disease (extra-ovarian spread). Nine patients with initial stage 1 disease developed metastases, and 20 patients had no evidence of recurrence over a mean follow-up period of 11.1 years. There was no significant correlation between any of the proliferation indices or with clinical outcomes. CONCLUSIONS: These results suggest that proliferation assessment is of limited value in the pathological assessment of GCT. Future studies should carefully specify the methods of assessing cell proliferation to ensure a reliable comparison of results. PMID- 18430758 TI - Antiangiogenic and vascular-disrupting agents in endometriosis: pitfalls and promises. AB - It is widely known that angiogenesis plays a key role in endometriotic lesion formation and development. Antiangiogenic treatments aimed at inhibiting new vessel formation have proven efficient in experimental models. However, as antiangiogenic strategies do not target pre-existing pericyte-protected vessels, they require chronic administration and are likely to be beneficial for early stage disease only or to prevent recurrence after surgery. Moreover, they may have detrimental effects on reproductive function. Vascular-disrupting agents (VDAs) have emerged as a promising new tool for the treatment of tumors. VDAs target established blood vessels, resulting in tumor ischemia and necrosis. These agents may therefore be more efficient against advanced disease. Two major types of VDAs are being developed for cancer: ligand-directed VDAs using antibodies, peptides and growth factors to deliver toxic effectors to tumor endothelium; and small-molecule VDAs exploiting physiological differences between tumor and normal endothelium to induce acute vascular shutdown. The ongoing evolution in genomics and proteomics is revolutionizing the discovery of novel endothelial markers. Several studies suggest that the vasculature of endometriotic lesions may have particular pathophysiological properties, which could be exploited for the development of selective VDAs. The aim of this review is to explore the merits and limitations of vascular therapy for the treatment of endometriosis. PMID- 18430759 TI - IFN-gamma-mediated extravillous trophoblast outgrowth inhibition in first trimester explant culture: a role for insulin-like growth factors. AB - Pre-eclampsia is often associated with inadequate cytotrophoblast invasion and remodelling of the uterine spiral arteries. Examining a first trimester, 2D in vitro explant culture model which mimics in vivo placentation, including trophoblast column formation and extravillous cytotrophoblast (EVT) migration, we previously suggested that excessive maternal decidual natural killer cell interferon (IFN-gamma) limits EVT migration. Types-1 and -2 insulin-like growth factor (IGF-1, IGF-2) are trophic for EVT, act through their surface receptors, IGFR-1 and IGFR-2, and are regulated by the IGF-binding proteins (IGFBPs). Could the observed IFN-gamma-mediated inhibition of EVT outgrowth and migration be related to either expression changes of IGF-1 or IGF-2, their receptors, their binding proteins, or apoptosis? Using the 2D explant culture model, we examined the effect of IFN-gamma exposure on IGF-1 and -2, IGFR-1 and -2, IGFBPs and apoptosis. IFN-gamma relatively increased IGF-1 and -2 secretion. In EVT, IFN gamma decreased IGFR-2, but not IGFR-1 expression. IGBP-2, -3 and -4 production were not influenced by IFN-gamma. IFN-gamma induced trophoblast apoptosis measured by the highly sensitive M30 neo-epitope, but not caspase 3 activity, in conditioned medium and EVT cell lysates. The observed IFN-gamma-mediated EVT migration inhibition may occur through the down-regulation of IGFR-2 and subtle induction of EVT apoptosis. PMID- 18430760 TI - Blunted increase of digital skin vasomotion following acetylcholine and sodium nitroprusside iontophoresis in systemic sclerosis patients. AB - OBJECTIVES: To test the hypothesis that finger skin vasomotion (FSV), a known factor influencing microvascular blood flow motion, is impaired in SSc patients. Possible relationships between FSV abnormalities and the severity and/or activity of SSc were also investigated. METHODS: FSV was investigated by means of spectral Fourier analysis of finger skin laser Doppler flowmetry (LDF) tracing, recorded before and following acetylcholine (ACh) or sodium nitroprusside (SNP) iontophoresis in 26 SSc patients and in 20 age-matched healthy controls. The power spectral density (PSD) of the 0.01-0.02, 0.02-0.06 and 0.06-0.2 Hz LDF oscillations (related to endothelial-, sympathetic- and myogenic-dependent FSV, respectively) was measured in PU(2) (perfusion units)/Hz. RESULTS: Compared with controls, SSc patients exhibited a significantly lower post-ACh and/or post-SNP percentage increase in PSD of 0.01-0.02 Hz (492 +/- 297% vs 283 +/- 167%; P < 0.005), of 0.02-0.06 Hz (336 +/- 205% vs 239 +/- 170%; P < 0.05) and of 0.06-0.2 Hz (223 +/- 91% vs 194 +/- 227%; P < 0.01) skin LDF oscillations. The post-SNP normalized PSD value of the 0.01-0.02 Hz and of the 0.02-0.06 Hz LDF oscillations was negatively related to SSc severity index (r = -0.407, P < 0.05 and r = -459, P < 0.05, respectively). CONCLUSIONS: This study showed a selective abnormality of the endothelial, sympathetic and myogenic-dependent FSV in SSc patients, consistent with a parallel endothelial, sympathetic and myogenic macrovascular dysfunction. This study also suggests a possible role of endothelial and sympathetic dysfunction in the progression of SSc. PMID- 18430761 TI - Predictors of psychological morbidity in parents of children with intellectual disabilities. AB - OBJECTIVE: This study examined predictors of excess psychological morbidity in parents of children with intellectual disabilities. METHODS: Thirty-two parents of children with intellectual disabilities and 29 parents of typically developing children completed the Hospital Depression and Anxiety Scale, and measures of social support, child problem behaviors, sleep quality, and perceived caregiver burden. RESULTS: Parents of children with intellectual disabilities registered high depression and anxiety scores, and the majority met the criteria for possible clinical depression and/or anxiety. The strongest predictor of psychological morbidity was caregiver burden. Analyses of its component dimensions indicated that feelings of guilt held the greatest consequence for depression and anxiety. CONCLUSIONS: Caregiver burden, in general, and its guilt component, in particular, predicted symptoms of depression and anxiety in parents of children with intellectual disabilities. Assisting such parents to resolve their feelings of guilt should benefit their psychological status. PMID- 18430762 TI - Evidence-based assessment of health-related quality of life and functional impairment in pediatric psychology. AB - OBJECTIVE: To provide an evidence-based review of measures of health-related quality of life (HRQOL) and functional impairment, with a specific focus on their use in the field of pediatric psychology. METHODS: As part of a larger survey of pediatric psychologists from the Society of Pediatric Psychology e-mail listserv (American Psychological Association, APA, Division 54), 16 measures were selected for this psychometric review. Measures that qualified for the review fell into one of the following three categories: (a) generic HRQOL scales, (b) disease specific quality of life scales, and (c) functional impairment rating scales. RESULTS: Psychometric characteristics (i.e., three types of reliability, two types of validity) were strong for the majority of measures reviewed, with 12 of the 16 measures meeting "well-established" evidence-based assessment criteria. Strengths and weaknesses of existing measures were noted. CONCLUSIONS: Recommendations for future work in this area of assessment are presented, including suggestions that further validation and exploration of measure properties such as factor analysis and changes in HRQOL over time be conducted. PMID- 18430763 TI - The use of statins and fate of small abdominal aortic aneurysms. AB - The aim of this study was to evaluate the value of statins in reducing abdominal aortic aneurysm (AAA) growth rate and improving freedom from aneurysm repair or rupture. One hundred and twenty-one patients with AAA undergoing ultrasonographic surveillance for at least one year were included in this retrospective study. Patients treated with statins had a decreased linear aneurysm growth rate than those not receiving statins (1.9+/-1.8 mm/year vs. 2.6+/-2.4 mm/year, P=0.27), but this difference did not reach statistical significance. Statin users had a better survival freedom from aneurysm repair or rupture (at 5 years: 72.3% vs. 52.5%, P=0.048). The impact of treatment with statins was even more evident in patients with a baseline aneurysm diameter<40 mm (at 5 years: 84.0% vs. 58.8%, P=0.022). When adjusted for age, coronary artery disease and baseline aneurysm diameter, treatment with statins had significantly better survival freedom from aneurysm repair or rupture (P=0.012, RR 0.34, 95% CI 0.14-0.78). The use of statins seems to slightly decrease the AAA growth rate and to significantly improve freedom from aneurysm repair and rupture. PMID- 18430764 TI - Hypereosinophilic syndrome and myocardial fibrosis. AB - A 62-year-old man was accepted to our institution because of hypereosinophilia, severe tricuspid regurgitation and isolated right restrictive myocardiopathy, with thrombi inside the right atrium and ventricle. Based on the diagnosis of hypereosinophilic syndrome plus eosinophilic myocarditis, the patient underwent a tricuspid valve repair and endomyocardiectomy. We briefly discuss hypereosinophilic syndrome myocardiopathy, and its management. PMID- 18430765 TI - Low replacement doses of thyroxine during food restriction restores type 1 deiodinase activity in rats and promotes body protein loss. AB - During food restriction, decreased basal metabolic rate secondary to reduced serum thyroid hormones levels contributes to weight loss resistance. Thyroxine (T(4)) and 3,3',5-tri-iodothyronine (T(3)) administration during caloric restriction produce deleterious side effects; however, the administration of physiological doses of T(4) during food restriction has never been evaluated. The aim of this study was to analyze the effects of low replacement doses of T(4) in Wistar rats subjected to 40% food restriction. Food restriction for 30 days led to significantly reduced liver type 1 deiodinase activity, serum TSH, leptin, T(4), T(3), metabolic rate, and body mass. The significant reduction in hepatic deiodinase activity found during food restriction was normalized in a dose dependent manner by T(4) replacement, showing that decreased type 1 deiodinase (D1) activity is secondary to decreased serum thyroid hormone levels during caloric restriction. The lowest replacement dose of T(4) did not normalize resting metabolic rate, but was able to potentiate the effects of food restriction on carcass fat loss and did not spare body protein. The highest dose of T(4) produced a normalization of daily oxygen consumption and determined a significant reduction in both carcass fat and protein content. Our results show that serum T(4) normalization during food restriction restores serum T(3) and liver D1 activity, while body protein is not spared. Thus, decreased serum T(4) during caloric restriction corresponds to a protective mechanism to avoid body protein loss, highlighting the importance of other strategies to reduce body mass without lean mass loss. PMID- 18430766 TI - Effects of early gestation GH administration on placental and fetal development in sheep. AB - Ovine GH (oGH) is synthesized in placental tissue during maximal placental growth and development. Our objectives were to localize oGH mRNA in the placenta, and study the impact of exogenous GH on twin pregnancies during the normal window (35 55 days of gestational age; dGA) of placental expression. In situ hybridization localized oGH mRNA in uterine luminal epithelium but not in tissues of fetal origin. While maternal GH and IGF-I concentrations were increased (P<0.001) approximately tenfold, uterine, uterine fluid, placental, and fetal weights were unaffected by treatment at either 55 or 135 dGA. Fetal length, liver weight, and liver weight per kg of body weight were unaffected by maternal GH treatment. However, in the cotyledon, IGF-binding protein (BP)-1 and IGFBP-4 mRNA concentrations were increased (P<0.05), while IGFBP-2 mRNA was decreased (P<0.05). The concentration of mRNA for IGFBP-3 was unaffected by treatment. Within the caruncle, IGFBP-1 mRNA was decreased (P<0.05), while IGFBP-3 and IGFBP 4 mRNA were increased (P<0.05), and IGFBP-2 mRNA was unchanged due to GH treatment. While our data indicate that elevated maternal GH and IGF-I concentrations during early and mid-gestation do not enhance placental and fetal growth in twin pregnancies, localization of GH mRNA in uterine luminal epithelium could explain GHs transitory expression from 35 to 55 dGA, since by the end of this period the majority of the uterine luminal epithelium has fused with chorionic binucleate cells forming the placental syncytium. PMID- 18430767 TI - Granule-bound starch synthase I is responsible for biosynthesis of extra-long unit chains of amylopectin in rice. AB - A rice Wx gene encoding a granule-bound starch synthase I (GBSSI) was introduced into the null-mutant waxy (wx) rice, and its effect on endosperm starches was examined. The apparent amylose content was increased from undetectable amounts for the non-transgenic wx cultivars to 21.6-22.2% of starch weight for the transgenic lines. The increase was in part due to a significant amount of extra long unit chains (ELCs) of amylopectin (7.5-8.4% of amylopectin weight), that were absent in the non-transgenic wx cultivars. Thus, actual amylose content was calculated to be 14.9-16.0% for the transgenic lines. Only slight differences were found in chain-length distribution for the chains other than ELCs, indicating that the major effect of the Wx transgene on amylopectin structure was ELC formation. ELCs isolated from debranched amylopectin exhibited structures distinct from amylose. Structures of amylose from the transgenic lines were slightly different from those of cv. Labelle (Wx(a)) in terms of a higher degree of branching and size distribution. The amylose and ELC content of starches of the transgenic lines resulted in the elevation of pasting temperature, a 50% decrease in peak viscosity, a large decrease in breakdown and an increase in setback. As yet undetermined factors other than the GBSSI activity are thought to be involved in the control of formation and/or the amount of ELCs. Structural analysis of the Wx gene suggested that the presence of a tyrosine residue at position 224 of GBSSI correlates with the formation of large amounts of ELCs in cultivars carrying Wx(a). PMID- 18430768 TI - Low-plasma insulin-like growth factor-I levels are associated with impaired endothelium-dependent vasodilatation in a cohort of untreated, hypertensive Caucasian subjects. AB - CONTEXT: Accumulating evidence suggests that IGF-I has protective vascular effects, supporting the possibility that IGF-I deficiency may contribute to atherosclerosis. However, the relationship between plasma IGF-I levels and endothelium-dependent vasodilatation is still unsettled. OBJECTIVE: We designed this present study to test the hypothesis that low-plasma IGF-I levels are associated with reduced endothelial function independently classical cardiovascular risk factors. SETTING: Outpatients were included in the study. PATIENTS: A total of 100 never-treated hypertensive Caucasian subjects participating in the CAtanzaro MEtabolic RIsk factors Study was recruited. INTERVENTIONS: Subjects underwent forearm blood flow (FBF) evaluation by strain gauge plethysmography in response to increasing doses of acetylcholine (ACh) (Sigma, Milan, Italy) and sodium nitroprusside (Malesci, Florence, Italy). Insulin sensitivity was estimated by the homeostasis model assessment index. RESULTS: Plasma IGF-I levels were significantly correlated with age (r = -0.300; P = 0.001), high-density lipoprotein serum cholesterol (r = 0.211; P = 0.017), homeostasis model assessment index (r = -0.355; P <0.0001), systolic blood pressure (r = -0.174; P = 0.042), glomerular filtration rate (r = 0.228; P = 0.011), and ACh-stimulated FBF (r = 0.565; P <0.0001). In a stepwise forward multivariate regression analysis, the strongest predictors of ACh-stimulated FBF response were plasma IGF-I levels, accounting for 31.9% of its variation. CONCLUSIONS: These results demonstrate, for the first time, that low-plasma IGF-I levels are highly associated with reduced endothelial function, an early step in atherogenesis process. PMID- 18430769 TI - High risk of hypopituitarism in patients who recovered from hemorrhagic fever with renal syndrome. AB - CONTEXT: Hemorrhagic fever with renal syndrome (HFRS) caused by hantaviruses, is a severe systemic infection, with acute shock, vascular leakage, hypotension, and acute renal failure. Pituitary ischemia/infarction and necrosis are known causes of hypopituitarism, often remaining unrecognized due to subtle clinical manifestations. Cases of hypopituitarism after HFRS were previously only sporadically reported. OBJECTIVE: The aim of this study was to determine, for the first time, the prevalence of hypopituitarism among HFRS survivors. SUBJECTS AND METHODS: In 60 adults (aged 35.8+/-1.3 yr) who recovered from HFRS 3.7 +/- 0.5 yr ago (median 2 yr), assessment of serum T(4), free T(4), TSH, IGF-I, prolactin, cortisol, and testosterone (in males) was followed by insulin tolerance test and/or GHRH+GH-releasing peptide-6 stimulation tests. RESULTS: Severe GH deficiency was confirmed in eight of 60 patients (13.3%): in five with multiple pituitary hormone deficiencies (MPHDs) and isolated in three. Thyroid axis deficiency was confirmed in five of 60 patients (8.3%), all with MPHD. Hypothalamus-pituitary-adrenal axis deficiency was observed in six of 60 (10.0%); in five with MPHD and isolated in one. Gonadal axis deficiency was confirmed in seven of 56 male subjects (12.5%): five with MPHD and isolated in two. Overall six patients (10.0%) had a single pituitary deficit (three GH, two gonadal, and one adrenal), and five (8.3%) had MPHD. The prevalence of patients having any endocrine deficiency was 18% (11 of 60). CONCLUSION: A high prevalence of hypopituitarism after recovery from HFRS is identified, with magnetic resonance imaging revealing atrophic pituitary and empty sella. Awareness is raised to neuroendocrine consequences of HFRS because unrecognized hypopituitarism significantly affects the physical and psychological well-being. PMID- 18430770 TI - Serum retinol binding protein 4 is related to insulin resistance and nonoxidative glucose metabolism in lean and obese women with normal glucose tolerance. AB - CONTEXT: Retinol-binding protein (RBP) 4 is secreted by adipose tissue and is postulated to be a determinant of insulin sensitivity. The mechanisms of RBP4 insulin desensitizing action remain unclear. OBJECTIVE: The aim of the present study was to estimate the relationships between serum RBP4 concentration with insulin sensitivity and oxidative and nonoxidative glucose metabolism in lean and obese women. DESIGN AND PARTICIPANTS: The study group consisted of 67 women with normal glucose tolerance, 27 lean and 40 overweight or obese. Insulin sensitivity was estimated with the euglycemic hyperinsulinemic clamp. Glucose and lipid oxidation was measured with indirect calorimetry in the basal state and during the last 30 min of the clamp. Nonoxidative glucose metabolism was calculated in insulin-stimulated conditions by subtracting glucose oxidation from total glucose metabolism. RESULTS: There was no difference in serum RBP4 concentration between lean and obese women. Serum RBP4 was inversely related to insulin sensitivity and nonoxidative glucose metabolism in the entire group (r = -0.36, P =0.003 in both cases) and within the subgroups of lean (r = -0.41, P =0.034 and r = -0.41, P =0.031) and obese women (r = -0.41, P =0.009 and r = -0.40, P =0.01, respectively). These relationships were independent of potential confounding factors. RBP4 levels were not associated with oxidative metabolism of glucose or lipid. CONCLUSIONS: Our data indicate that serum RBP4 is related to decreased insulin sensitivity, mostly through its association with nonoxidative glucose metabolism. PMID- 18430771 TI - Thyroid autoantibodies and thyroid function in subjects exposed to Chernobyl fallout during childhood: evidence for a transient radiation-induced elevation of serum thyroid antibodies without an increase in thyroid autoimmune disease. AB - CONTEXT: An increase in the prevalence of thyroid autoantibodies (ATAs) was reported 6-8 yr after the Chernobyl accident in radiation-exposed children and adolescents. OBJECTIVE: Our objective was to reassess the effects of childhood radiation exposure on ATAs and thyroid function 13-15 yr after the accident. DESIGN AND SETTING: We measured the antithyroglobulin (TgAbs) and antithyroperoxidase (TPOAbs) antibodies and TSH in 1433 sera collected between 1999 and 2001 from 13- to 17-yr-old adolescents born between January 1982 and October 1986 in paired contaminated and noncontaminated villages of Belarus, Ukraine, and Russia. A total of 1441 sera was collected from age- and sex-matched controls living in Denmark and Sardinia (Italy). Free T(4) and free T(3) were measured when TSH was abnormal. RESULTS: TPOAb prevalence was higher in contaminated than in noncontaminated Belarusian children (6.4 vs. 2.4%; P = 0.02) but lower than previously reported (11%) in a different contaminated Belarus village. No difference in TPOAb prevalence was found in Ukrainian and Russian villages. TgAbs showed no difference between contaminated and noncontaminated Belarus and Ukraine, whereas in Russia they showed a relative increase in the exposed subjects with respect to the unexposed, who showed an unexpectedly lower prevalence of TgAbs. Besides radiation exposure, female gender was the only variable significantly correlated with ATAs in all groups. ATA prevalence in nonexposed villages of Belarus, Ukraine, and Russian Federation did not differ from that found in Sardinia and Denmark. With few exceptions, thyroid function was normal in all study groups. CONCLUSIONS: TPOAb prevalence in adolescents exposed to radioactive fallout was still increased in Belarus 13-15 yr after the Chernobyl accident. This increase was less evident than previously reported and was not accompanied by thyroid dysfunction. Our data suggest that radioactive fallout elicited a transient autoimmune reaction, without triggering full-blown thyroid autoimmune disease. Longer observation periods are needed to exclude later effects. PMID- 18430772 TI - Influence of age at menarche on forearm bone microstructure in healthy young women. AB - BACKGROUND: Shorter estrogen exposure from puberty onset to peak bone mass attainment may explain how late menarche is a risk factor for osteoporosis. The influence of menarcheal age (MENA) on peak bone mass, cortical, and trabecular microstructure was studied in 124 healthy women aged 20.4 +/- 0.6 (sd) yr. METHODS: At distal radius, areal bone mineral density (aBMD) was measured by dual energy x-ray absorptiometry, and volumetric bone mineral density (BMD) and microstructure were measured by high-resolution peripheral computerized tomography, including: total, cortical, and trabecular volumetric BMD and fraction; trabecular number, thickness, and spacing; cortical thickness (CTh); and cross-sectional area (CSA). RESULTS: Median MENA was 12.9 yr. Mean aBMD T score of the whole cohort was slightly positive. aBMD was inversely correlated to MENA for total radius (R = -0.21; P = 0.018), diaphysis (R = -0.18; P = 0.043), and metaphysis (R = -0.19; P = 0.031). Subjects with MENA more than the median [LATER: 14.0 +/- 0.7 (+/-sd) yr] had lower aBMD than those with MENA less than the median (EARLIER: 12.1 +/- 0.7 yr) in total radius (P = 0.026), diaphysis (P = 0.042), and metaphysis (P = 0.046). LATER vs. EARLIER displayed lower total volumetric BMD (315 +/- 54 vs. 341 +/- 56 mg HA/cm(3); P = 0.010), cortical volumetric BMD (874 +/- 49 vs. 901 +/- 44 mg HA/cm(3); P = 0.003), and CTh (774 +/- 170 vs. 849 +/- 191 microm; P = 0.023). CTh was inversely related to CSA (R = -0.46; P < 0.001). In LATER reduced CTh was associated with 5% increased CSA. CONCLUSIONS: In healthy young adult women, a 1.9-yr difference in mean MENA was associated with lower radial aBMD T score, lower CTh without reduced CSA, a finding compatible with less endocortical accrual. It may explain how late menarche is a risk factor for forearm osteoporosis. PMID- 18430773 TI - Effects of short-term high-fat, high-energy diet on hepatic and myocardial triglyceride content in healthy men. AB - CONTEXT: An association has been suggested between elevated plasma nonesterified fatty acid (NEFA) levels, myocardial triglyceride (TG) accumulation, and myocardial function. OBJECTIVE: Our objective was to investigate the effects of an elevation of plasma NEFA by a high-fat, high-energy (HFHE) diet on hepatic and myocardial TG accumulation, and on myocardial function. DESIGN: There were 15 healthy males (mean +/- sd age: 25.0 +/- 6.6 yr) subjected to a 3-d HFHE diet consisting of their regular diet, supplemented with 800 ml cream (280 g fat) every day. METHODS: (1)H-magnetic resonance spectroscopy was performed for assessing hepatic and myocardial TGs. Furthermore, left ventricular function was assessed using magnetic resonance imaging. RESULTS: The HFHE diet increased hepatic TGs compared with baseline (from 2.01 +/- 1.79 to 4.26 +/- 2.78%; P = 0.001) in parallel to plasma TGs and NEFA. Myocardial TGs did not change (0.38 +/ 0.18 vs. 0.40 +/- 0.12%; P = 0.7). The HFHE diet did not change myocardial systolic function. Diastolic function, assessed by dividing the maximum flow across the mitral valve of the early diastolic filling phase by the maximum flow of the atrial contraction (E/A ratio), decreased compared with baseline (from 2.11 +/- 0.39 to 1.89 +/- 0.33; P = 0.031). This difference was no longer significant after adjustment for heart rate (P = 0.12). CONCLUSIONS: Short-term HFHE diet in healthy males results in major increases in plasma TG and NEFA concentrations and hepatic TGs, whereas it does not influence myocardial TGs or myocardial function. These observations indicate differential, tissue-specific partitioning of TGs and/or fatty acids among nonadipose organs during HFHE diet. PMID- 18430774 TI - Hyperinsulinemia induces insulin resistance on glucose and lipid metabolism in a human adipocytic cell line: paracrine interaction with myocytes. AB - CONTEXT: Adipocytes release a variety of factors which deregulation could provide the basis for complications such as insulin resistance, an early defect on the onset of type 2 diabetes. Such insulin resistance can initially be overcome by compensatory hyperinsulinemia, but the prolonged presence of the hormone can be detrimental for insulin sensitivity. OBJECTIVE: The objective of the study was to dissect the molecular mechanisms that may regulate hyperinsulinemia-induced insulin resistance in a human liposarcoma cell line and its paracrine interactions with a human rhabdomyosarcoma cell line. DESIGNS: We studied glucose uptake, lipolysis, insulin signaling, and secretion pattern at different days of adipocyte differentiation in the presence of insulin. RESULTS: Adipocytes differentiated for 14 d gain insulin sensitivity on glucose uptake and inhibition of lipolysis, but prolonged cultures develop an insulin-resistant state characterized by an increase in phosphatase and tensin homolog-deleted on chromosome 10 expression and defects in insulin signaling at the insulin receptor substrate-1/AKT level. The secretion pattern of nonesterified fatty acids, IL-6, adiponectin, leptin, and monocyte chemotactic protein-1 was in keeping with the changes in insulin sensitivity during differentiation. An inverse biphasic response was also observed in human myocytes when they were cultured with various adipocyte-conditioned media, although insulin resistance was detected earlier than in adipocytes. This behavior mimics hyperinsulinemia because insulin action was restored when adipocytes were cultured in the absence of the hormone. Pharmacological treatment of adipocytes with a liver X receptor agonist reestablishes insulin-stimulated glucose uptake, whereas treatment with a peroxisome proliferator-activated receptor-gamma agonist restored the antilipolytic action of insulin. CONCLUSIONS: Hyperinsulinemia deregulates adipocyte secretion pattern, producing insulin resistance in adipocytes and myocytes, a situation that can be ameliorated with nuclear receptor agonists. PMID- 18430775 TI - Cardiac status after childhood growth hormone treatment of Turner syndrome. AB - CONTEXT: In Turner syndrome (TS), GH treatment is well established. Data on cardiac status after discontinuation of treatment are scarce. This study aimed to assess biventricular size and function in TS at least 6 months after discontinuation of GH treatment. METHODS: TS patients and healthy women prospectively underwent cardiac magnetic resonance imaging. Ventricular two dimensional tomographic cine data were acquired to obtain biventricular volume, mass, and ejection fraction. Atrioventricular valve flow measurements were performed using a two-dimensional flow-sensitized sequence. Flow velocity curves were calculated and indices of biventricular diastolic filling were derived. RESULTS: Thirty-one patients [mean (sd) age 20 (2) yr, body surface area 1.75 (0.15) m(2), 5 (2) yr after GH discontinuation] and 23 normal control women [age 21 (2) yr, body surface area 1.80 (0.13) m(2)] were included. Compared with controls, patients had smaller mean end-diastolic volumes [right ventricle (RV), 84 (11) ml/m(2) vs. 79 (10), P = 0.02; left ventricle (LV), 81 (10) vs. 72 (9), P < 0.001], end-systolic volumes [RV 38 (7) ml/m(2) vs. 36 (6), P = 0.04; LV 34 (5) vs. 29 (4), P < 0.001], and stroke volumes [RV 46 (6) ml/m(2) vs. 43 (6), P = 0.03; LV, 47 (7) vs. 44 (4), P = 0.02]. Patients had a higher mean heart rate [79 (13) beats/min vs. 71 (10), P < 0.05]. Biventricular ejection fraction, mass, cardiac output, and diastolic filling pattern were comparable. CONCLUSION: After discontinuation of GH treatment TS patients showed no myocardial hypertrophy and well-preserved biventricular function. Ventricular volumes were smaller in Turner patients, compared with controls, whereas mean heart rate was higher. These last observations may be part of the natural development in TS and not linked to GH treatment, which at this point we consider safe. PMID- 18430776 TI - Interleukin-6 markedly decreases skeletal muscle protein turnover and increases nonmuscle amino acid utilization in healthy individuals. AB - CONTEXT: IL-6 is a key modulator of immune function and suggested to be involved in skeletal muscle wasting as seen in sepsis. OBJECTIVE: Our objective was to determine the role of IL-6 in human in vivo systemic and skeletal muscle amino acid metabolism and protein turnover. SUBJECTS AND METHODS: There were 12 healthy men infused for 3 h with saline (saline, n = 6) or recombinant human IL (rhIL)-6 (n = 6). Systemic and muscle protein turnover was determined with a combination of tracer dilution methodology, primed constant infusion of L-[ring (2)H(5)]phenylalanine, and femoral arterial-venous blood differences and m. vastus lateralis biopsies after 2-h basal, 3-h infusion, and 3 h after infusion. RESULTS: The IL-6 concentration after 30-min infusion was approximately 4 (saline) and 140 pg/ml (rhIL-6). Three-hour rhIL-6 infusion caused an approximate 50% decrease in muscle protein turnover, albeit synthesis was more suppressed than breakdown, causing a small increase in net muscle protein breakdown. Furthermore, rhIL-6 decreased arterial amino acid concentration with 20-40%, despite the increase net release from muscle. CONCLUSIONS: We demonstrated that IL-6 profoundly alters amino acid turnover. A substantial decrease in plasma amino acids was observed with a concomitant 50% decrease in muscle protein turnover, however, modest increase in net muscle degradation. We hypothesize that the profound reduction in muscle protein turnover and modest increase in net degradation are primarily caused by the reduced plasma amino acid availability and not directly mediated by IL-6. PMID- 18430777 TI - A novel variant of familial glucocorticoid deficiency prevalent among the Irish Traveler population. AB - CONTEXT: Familial glucocorticoid deficiency (FGD) is an autosomal recessive disorder characterized by distinct clinical, biochemical, and genetic abnormalities. The prevalence of FGD is unknown, with the likelihood that cases remain undiagnosed. We noted a significant proportion of our FGD cases are Irish Travelers. Irish Travelers are an endogamous nomadic group ethnically and genetically distinct from Roma gypsies. AIMS: The objective of the study was to describe the clinical features and assess the prevalence of FGD amongst Irish Travelers in the Republic of Ireland and describe their phenotype. METHODS: Diagnosis of FGD was based on clinical features, high ACTH, and low cortisol concentrations with normal renin and aldosterone concentrations and exclusion of other causes of adrenal failure. Data from the Republic of Ireland Census 2006 were used. RESULTS: We identified 21 cases of FGD, generating an overall prevalence of one in 201,898. We report nine Irish Travelers (five females) with FGD related to a new gene negative for melanocortin-2 receptor and melanocortin-2 receptor accessory protein mutations. Of a total population of 22,557 Travelers, this yields a disease prevalence of one in 2506 with a carrier frequency of one in 25 in this group and represents a prevalence of one in 665 and a carrier frequency of one in 13 in the 4- to 15-yr Traveler age group. All nine children had a later onset of FGD due to the fact that their initial investigations revealed normal cortisol (422-575 nmol/liter) and ACTH (<34 ng/liter) concentrations. CONCLUSION: We report a high prevalence of FGD among Irish Travelers. Their subtle phenotype and initial normal biochemistry may delay the early diagnosis of FGD. PMID- 18430778 TI - Effect of weight loss by gastric bypass surgery versus hypocaloric diet on glucose and incretin levels in patients with type 2 diabetes. AB - CONTEXT: Gastric bypass surgery (GBP) results in rapid weight loss, improvement of type 2 diabetes (T2DM), and increase in incretins levels. Diet-induced weight loss also improves T2DM and may increase incretin levels. OBJECTIVE: Our objective was to determine whether the magnitude of the change of the incretin levels and effect is greater after GBP compared with a low caloric diet, after equivalent weight loss. DESIGN AND METHODS: Obese women with T2DM studied before and 1 month after GBP (n = 9), or after a diet-induced equivalent weight loss (n = 10), were included in the study. Patients from both groups were matched for age, body weight, body mass index, diabetes duration and control, and amount of weight loss. SETTING: This outpatient study was conducted at the General Clinical Research Center. MAIN OUTCOME MEASURES: Glucose, insulin, proinsulin, glucagon, gastric inhibitory peptide (GIP), and glucagon-like peptide (GLP)-1 levels were measured after 50-g oral glucose. The incretin effect was measured as the difference in insulin levels in response to oral and to an isoglycemic iv glucose load. RESULTS: At baseline, none of the outcome variables (fasting and stimulated values) were different between the GBP and diet groups. Total GLP-1 levels after oral glucose markedly increased six times (peak:17 +/- 6 to 112 +/- 54 pmol/liter; P < 0.001), and the incretin effect increased five times (9.4 +/- 27.5 to 44.8 +/- 12.7%; P < 0.001) after GBP, but not after diet. Postprandial glucose levels (P = 0.001) decreased more after GBP. CONCLUSIONS: These data suggest that early after GBP, the greater GLP-1 and GIP release and improvement of incretin effect are related not to weight loss but rather to the surgical procedure. This could be responsible for better diabetes outcome after GBP. PMID- 18430779 TI - A single point mutation in the LN domain of LAMA2 causes muscular dystrophy and peripheral amyelination. AB - Mutations in the gene encoding the basal lamina (BL) component laminin alpha2 (LAMA2) cause merosin-deficient congenital muscular dystrophy 1A (MDC1A), a complex disorder that includes hypomyelination and myodegeneration. In dystrophia muscularis (dy) mice bearing Lama2 mutations, myofibers and Schwann cells fail to assemble stable BLs, which are thought to be crucial for myofiber survival and Schwann cell differentiation. Here, we describe defects in a new allele of Lama2 in mice, nmf417, in which a point mutation substitutes Arg for Cys79 at a universally conserved CxxC motif in the laminin N-terminal (LN) domain; this domain mediates laminin-laminin interactions. nmf417 homozygosity caused progressive myodegeneration and severe peripheral amyelination in nerve roots, similar to previous Lama2 mutations, but without the pervasive BL thinning previously associated with the disorder. In direct contrast to the previously characterized dy and dy2J alleles, nmf417 homozygous myofibers frequently had thickened BLs. Severe amyelination in nmf417-mutant nerve roots suggested complete laminin 2 inactivation for Schwann cells, although myelinated fibers had normal BLs. The results reveal crucial roles for the LN domain CxxC motif in both nerve and muscle, but challenge expected relationships between LN-domain function, Ln2 activity and BL stability. The nmf417 mutation provides a defined animal model in which to investigate mechanisms and treatments for moderate forms of MDC1A. PMID- 18430780 TI - Analysis of Fyn function in hemostasis and alphaIIbbeta3-integrin signaling. AB - Recent studies have shown that Src-family kinases (SFKs) play an important role in mediating integrin signalling, and the beta3 subunit of alphaIIbbeta3 integrin has been shown to interact with multiple SFK members. Here, we analyzed the interactions and functional consequences of Fyn and Src binding to alphaIIbbeta3. Fyn associated with the beta3 subunit in resting and thrombin-aggregated platelets, whereas interaction between Src and alphaIIbbeta3 was seen predominantly in resting but not in thrombin-aggregated platelets. We have also observed that Fyn but not Src localized to focal adhesions in CHO cells adherent to fibrinogen through alphaIIbbeta3. On the basis of these differences, we wanted to determine the sequence requirements for the interaction of Fyn and Src within the beta3-cytoplasmic domain. Whereas Src association required the C-terminal region of beta3, Fyn continued to interact with mutants that could no longer associate with Src and that contained as few as 13 membrane-proximal amino acids of the beta3-cytoplasmic tail. Using deletion mutants of beta3-cytoplasmic tails expressed as GST-fusion proteins, we narrowed down the Fyn-binding site even further to the amino acid residues 721-725 (IHDRK) of the beta3-cytoplasmic domain. On the basis of these observations, we explored whether Fyn-/- mice exhibited any abnormalities in hemostasis and platelet function. We found that Fyn-/- mice significantly differed in their second bleeding times compared with wild-type mice, and platelets from Fyn-/- mice exhibited delayed spreading on fibrinogen-coated surfaces. Using mutant forms of Fyn, it appears that its kinase activity is required for its localization to focal adhesions and to mediate alphaIIbbeta3-dependent cell spreading. Our results suggest that Fyn and Src have distinct requirements for interaction with alphaIIbbeta3; and, consequently, the two SFK can mediate different functional responses. PMID- 18430781 TI - Rab5 modulates aggregation and toxicity of mutant huntingtin through macroautophagy in cell and fly models of Huntington disease. AB - Huntington disease (HD) is caused by a polyglutamine-expansion mutation in huntingtin (HTT) that makes the protein toxic and aggregate-prone. The subcellular localisation of huntingtin and many of its interactors suggest a role in endocytosis, and recently it has been shown that huntingtin interacts indirectly with the early endosomal protein Rab5 through HAP40. Here we show that Rab5 inhibition enhanced polyglutamine toxicity, whereas Rab5 overexpression attenuated toxicity in our cell and fly models of HD. We tried to identify a mechanism for the Rab5 effects in our HD model systems, and our data suggest that Rab5 acts at an early stage of autophagosome formation in a macromolecular complex that contains beclin 1 (BECN1) and Vps34. Interestingly chemical or genetic inhibition of endocytosis also impeded macroautophagy, and enhanced aggregation and toxicity of mutant huntingtin. However, in contrast to Rab5, inhibition of endocytosis by various means suppressed autophagosome-lysosome fusion (the final step in the macroautophagy pathway) similar to bafilomycin A1. Thus, Rab5, which has previously been thought to be exclusively involved in endocytosis, has a new role in macroautophagy. We have previously shown that macroautophagy is an important clearance route for several aggregate-prone proteins including mutant huntingtin. Thus, better understanding of Rab5 regulated autophagy might lead to rational therapeutic targets for HD and other protein-conformation diseases. PMID- 18430782 TI - Drosophila melanogaster kl-3 and kl-5 Y-loops harbor triple-stranded nucleic acids. AB - Primary spermatocyte nuclei of Drosophila melanogaster contain three prominent lampbrush-like loops. The development of these structures has been associated with the transcription of three fertility factors located on the Y chromosome, named kl-5, kl-3 and ks-1. These loci have huge physical dimensions and contain extremely long introns. In addition, kl-3 and kl-5 were shown to encode two putative dynein subunits required for the correct assembly of the sperm axoneme. Here, we show that both the kl-5 and kl-3 loops are intensely decorated by monoclonal antibodies recognizing triple-stranded nucleic acids, and that each loop presents a peculiar molecular organization of triplex structures. Moreover, immunostaining of Drosophila hydei primary spermatocytes revealed that also in this species - which diverged from D. melanogaster 58 million years ago - Y-loops are decorated by anti-triplex antibodies, strongly suggesting a conserved role of loop-associated triplexes. Finally, we showed that in D. melanogaster wild-type lines that are raised at the non-permissive temperature of 31+/-0.5 degrees C (which is known to induce male sterility in flies) both the triplex immunostaining and the axonemal dynein heavy chains encoded by kl-3 and kl-5 are no longer detectable, which suggests a functional correlation between loop associated triplexes, the presence of axonemal proteins and male fertility in fly. PMID- 18430783 TI - Zfp64 participates in Notch signaling and regulates differentiation in mesenchymal cells. AB - Notch signaling is required for multiple aspects of tissue and cell differentiation. In this study, we identified zinc finger protein 64 (Zfp64) as a novel coactivator of Notch1. Zfp64 is associated with the intracellular domain of Notch1, recruited to the promoters of the Notch target genes Hes1 and Hey1, and transactivates them. Zfp64 expression is under the control of Runx2, and is upregulated by direct transactivation of its promoter. Zfp64 suppresses the myogenic differentiation of C2C12 cells and promotes their osteoblastic differentiation. Our data demonstrate two functions of Zfp64: (1) it is a downstream target of Runx2 and, (2) its cognate protein acts as a coactivator of Notch1, which suggests that Zfp64 mediates mesenchymal cell differentiation by modulating Notch signaling. PMID- 18430784 TI - In vivo role of lipid adducts on Wingless. AB - Two lipids (palmitate and palmitoleic acid) are appended onto Wnt proteins. It has been suggested that palmitate is required for signalling, whereas palmitoleic acid is necessary for progression through the secretory pathway. By mutating the relevant amino acids, we have investigated how these adducts contribute to the secretion and signalling activity of Wingless, the main Drosophila member of the Wnt family. Analysis of Wingless with a Cysteine 93 to Alanine mutation ([C93A]Wingless) shows that palmitoylation is essential for signalling activity in vivo (as well as in cultured cells). Moreover, without palmitate, Wingless fails to reach the surface of imaginal disc cells and, as electron microscopy (EM) analysis suggests, appears to accumulate in the endoplasmic reticulum (ER). Artificial targeting of palmitate-deficient Wingless to the plasma membrane does not rescue signalling activity. Therefore, palmitate at C93 has a dual role: in secretion and signalling. From our analysis of [S239A]Wingless, which lacks a conserved residue shown to be acylated in Wnt3a, we infer that palmitoleic acid is not, as previously suggested, absolutely required for secretion. Nevertheless, this mutant has poor signalling activity, suggesting that palmitoleic acid contributes significantly to signalling. We suggest that the overall level of lipidation affects signalling activity. PMID- 18430785 TI - Differential regulation of naive and memory CD4+ T cells by alternatively activated dendritic cells. AB - Promising immunotherapeutic tools for T cell-mediated pathologies are alternatively activated dendritic cells (aaDC), which exert their effect through the regulation and tolerization of T cells. As naive and memory T cells have different susceptibilities to tolerogenic signals, it is important to understand the modulatory effects of aaDC on these T cell subsets. We have examined regulation of naive and memory CD4+ T cells by human aaDC generated with dexamethasone, the active form of vitamin D3, 1alpha,25-dihydroxyvitamin D3, and LPS. Although aaDC induced low, primary, allogeneic responses by naive and memory T cells, aaDC regulated the differentiation of these T cell subsets in a distinct manner. Naive T cells primed by aaDC retained a strong, proliferative capacity upon restimulation but were skewed toward a low IFN-gamma/high IL-10 cytokine profile. In contrast, memory T cells primed by aaDC became hyporesponsive in terms of proliferation and cytokine production. Induction of anergy in memory T cells by aaDC was not a result of the presence of CD25hi regulatory T cells and could be partially reversed by IL-2. Both T cell subsets acquired regulatory activity and inhibited primary CD4 and CD8 responses. Addition of exogenous IL 12p70 during T cell priming by aaDC prevented anergy induction in memory T cells and cytokine polarization in naive T cells, indicating that the lack of IL-12p70 is a key feature of aaDC. Our finding that aaDC differentially regulate naive and memory T cells is important for understanding and maximizing the therapeutic potential of aaDC. PMID- 18430786 TI - Subversion of complement activation at the bacterial surface promotes serum resistance and opsonophagocytosis of Francisella tularensis. AB - Francisella tularensis (Ft) is resistant to serum but requires complement factor C3-derived opsonins for uptake by phagocytic cells and subsequent intracellular growth. In this study, we show that C3 fragments, deposited on Ft, are detected by anti-C3d and -iC3b mAb and that the classical and the alternative pathways are involved in this event. This was demonstrated using C2-depleted sera and specific inhibitors of the classical-versus-alternative pathways of complement activation. Further, we demonstrate that factor C4b, which is crucial for the classical pathway, is deposited on the surface of Ft. In contrast, the C5b-C9 membrane attack complex (MAC) is not assembled on the surface of Ft, which may explain its resistance to complement killing. Deposition of C3 opsonins leads to enhanced phagocytosis by human immature dendritic cells (DC), which leads to intracellular survival, growth, and DC death. Finally, we show that factor H (fH) can bind to the surface of Ft. We believe our data suggest that important virulence factors for Ft are its ability to bind fH and inactivate C3b to iC3b, which culminates in opsonin-induced uptake for subsequent intracellular growth. C3b inactivation also leads to inefficient MAC assembly, which contributes to the ability of this bacterium to resist complement lysis. PMID- 18430788 TI - Activation of 9-[(R)-2-[[(S)-[[(S)-1-(Isopropoxycarbonyl)ethyl]amino] phenoxyphosphinyl]-methoxy]propyl]adenine (GS-7340) and other tenofovir phosphonoamidate prodrugs by human proteases. AB - 9-[(R)-2-[[(S)-[[(S)-1-(Isopropoxycarbonyl)ethyl]amino] phenoxyphosphinyl] methoxy]propyl]adenine (GS-7340) is an isopropylalaninyl phenyl ester prodrug of the nucleotide HIV reverse transcriptase inhibitor tenofovir (TFV; 9-[(2 phosphonomethoxy)propyl]adenine) exhibiting potent anti-HIV activity and enhanced ability to deliver parent TFV into peripheral blood mononuclear cells (PBMCs) and other lymphatic tissues in vivo. The present study focuses on the intracellular metabolism of GS-7340 and its activation by a variety of cellular hydrolytic enzymes. Incubation of human PBMCs in the presence of GS-7340 indicates that the prodrug is hydrolyzed slightly faster to an intermediate TFV-alanine conjugate (TFV-Ala) in quiescent PBMCs compared with activated cells (0.21 versus 0.16 pmol/min/10(6) cells). In contrast, the conversion of TFV-Ala to TFV and subsequent phosphorylation to TFV-diphosphate occur more rapidly in activated PBMCs. The activity of GS-7340 hydrolase producing TFV-Ala in PBMCs is primarily localized in lysosomes and is sensitive to inhibitors of serine hydrolases. Cathepsin A, a lysosomal serine protease has recently been identified as the primary enzyme activating GS-7340 in human PBMCs. Results from the present study indicate that in addition to cathepsin A, a variety of serine and cysteine proteases cleave GS-7340 and other phosphonoamidate prodrugs of TFV. The substrate preferences displayed by these enzymes toward TFV amidate prodrugs are nearly identical to their preferences displayed against oligopeptide substrates, indicating that GS-7340 and other phosphonoamidate derivatives of TFV should be considered peptidomimetic prodrugs of TFV. PMID- 18430787 TI - Synthetic oligonucleotides as modulators of inflammation. AB - Synthetic oligodeoxynucleotides (ODN) containing unmethylated CpG motifs mimic the immunostimulatory activity of bacterial DNA. CpG ODN directly stimulate human B cells and plasmacytoid dendritic cells, promote the production of Th1 and proinflammatory cytokines, and trigger the maturation/activation of professional APC. CpG ODN are finding use in the treatment of cancer, allergy, and infection. In contrast, ODN containing multiple TTAGGG motifs mimic the immunosuppressive activity of self-DNA, down-regulating the production of proinflammatory and Th1 cytokines. Preclinical studies suggest that "suppressive" ODN may slow or prevent diseases characterized by pathologic immune stimulation, including autoimmunity and septic shock. Extensive studies in animal models suggest that the therapeutic value of CpG and TTAGGG ODN may be optimized by early administration. PMID- 18430789 TI - Carmustine-resistant cancer cells are sensitized to temozolomide as a result of enhanced mismatch repair during the development of carmustine resistance. AB - The cytotoxicity of the monofunctional alkylator temozolomide (TMZ) is mediated by mismatch repair (MMR) triggered by O(6)-alkylguanine, whereas MMR protects cells against bifunctional alkylators, including carmustine (BCNU). Therefore, TMZ may be cytotoxic to BCNU-resistant cancer cells because MMR affects sensitivity to TMZ and BCNU in a converse way. We evaluated TMZ cytotoxicity on BCNU-resistant variant (CEM-R) compared with the parental CCRF-CEM cell line (CEM S). The mechanisms of its BCNU-resistance involved DNA repairs including nucleotide excision repair, base excision repair, alkylguanine alkyltransferase, MMR, and apoptotic and survival pathways. In particular, transcript levels of MMR related hMLH1 and hMSH2 were enhanced in CEM-R cells. CEM-R cells were 8-fold more BCNU-resistant but surprisingly 9-fold more TMZ-sensitive than were CEM-S cells. Although TMZ-induced adducts include N-alkylated purines and O(6) alkylguaine, DNA excision repair was enhanced in CEM-R cells, suggesting the efficient repair of N-alkylation adducts. Cotreatment with methoxyamine, a base excision repair inhibitor, did not sensitize CEM-R cells to TMZ, suggesting little or no contribution of N-alkylation to TMZ-induced cytotoxicity. Cotreatment with O(6)-benzylguanine, an alkylguanine alkyltransferase inhibitor, further sensitized CEM-R cells to TMZ, confirming the cytotoxic impact of O(6) alkylguanine. Cotreatment with cadmium chloride, an MMR inhibitor, disrupted the sensitivity of CEM-R cells to TMZ. The sensitivity to TMZ was reversed in the CEM R variant clone that had been established by transfecting CEM-R cells with short hairpin hRNA against hMLH1, suggesting the critical role of MMR on sensitization to TMZ. In conclusion, BCNU-resistant CEM-R cells were sensitized to TMZ as a result of enhanced MMR during the development of BCNU resistance. PMID- 18430790 TI - Insulinoma cell calcium-sensing receptor influences insulin secretion in a case with concurrent familial hypocalciuric hypercalcemia and malignant metastatic insulinoma. AB - CONTEXT AND OBJECTIVE: Arterial stimulation and venous sampling (ASVS) is an important technique for localizing insulinoma. The principle behind ASVS is that insulin secretion is promoted from insulinoma cells by the injection of calcium into the insulinoma-feeding artery. However, the mechanism for ASVS-induced insulin secretion remains unclear. Both insulinoma and familial hypocalciuric hypercalcemia (FHH) are rare diseases. This study reports on a case in which both of these diseases occur concurrently. DESIGN AND PATIENT: The patient with FHH also suffered from insulinoma. We reasoned that insulin secretion for ASVS is dependent on the calcium-sensing receptor (CaSR). ASVS was performed on this patient. The expression of the CaSR protein and corresponding mRNA were confirmed. RESULTS: No significant changes in the plasma levels of insulin and C peptide were observed during ASVS. The patient was clinically diagnosed as having FHH. We confirmed that a mutation in the CaSR gene was present in the genomic DNA of this patient and that there were no mutations in the multiple endocrine neoplasia type 1 gene. In addition, expression of both CaSR mRNA and CaSR protein was confirmed in the insulinoma samples. CONCLUSION: These results suggest that the CaSR gene is involved in ASVS-induced insulin secretion. PMID- 18430791 TI - Confirmed beta16 Arg/Arg polymorphism in a patient with uncontrolled asthma. AB - OBJECTIVE: To report a case of confirmed beta(16) Arg/Arg polymorphism (Arg/Arg) in a patient with uncontrolled asthma. CASE SUMMARY: A 49-year-old black female presented to the emergency department with acute shortness of breath with subsequent intubation. After extubation, she reported multiple hospitalizations for asthma with one prior intubation, adherence to asthma medications, and very frequent use of her short-acting beta(2)-agonist (SABA). Because of her asthma history, self-reported adherence, and race, she was tested for beta(2) adrenoreceptor genotype, which revealed Arg/Arg. Based on these findings, beta(2) agonists were discontinued and tiotropium (maintenance) and ipratropium (primary rescue) were initiated as part of her asthma regimen. Application of the Naranjo probability scale revealed probable causality between uncontrolled asthma in our patient and SABA use. The patient is followed in our outpatient pulmonary clinic and, at time of writing, had not been admitted to our hospital for asthma-related events. DISCUSSION: Approximately 15% of Americans with asthma are Arg/Arg, with an increased prevalence in black and Asian populations. It is hypothesized that changes in the degree of sensitivity or desensitization to the bronchodilator effect of beta(2)-agonists may occur in these individuals. Data exist, although they are conflicting, suggesting that inhaled beta(2)-agonists may worsen clinical outcomes. Trials have reported declines in peak expiratory flow rates plus increases in asthma symptoms and exacerbations when SABAs have been used regularly in patients with Arg/Arg. Studies evaluating long-acting beta(2) agonists (LABAs) have inconsistent results. Preliminary data suggest that anticholinergics may serve as a beneficial primary rescue medication instead of beta(2)-agonists in patients with Arg/Arg. CONCLUSIONS: Clinicians should be aware of factors (eg, race and polymorphisms) that may predict unfavorable outcomes with regular SABA and possibly LABA use. Patients with poor asthma control despite adherence to asthma therapy may benefit from beta(2) adrenoreceptor genotyping and, possibly, from anticholinergics. PMID- 18430792 TI - Rifamycin antibiotics for treatment of Clostridium difficile-associated diarrhea. AB - OBJECTIVE: To review the existing data on use of the rifamycin class of antibiotics as therapy for Clostridium difficile-associated diarrhea (CDAD). DATA SOURCES: A literature search was performed using PubMed (1996-January 2008), abstracts from the International Conference on Antimicrobial Agents and Chemotherapy (September 2007), the Infectious Diseases Society of America (October 2007), Salix Pharmaceuticals Web site (January 2008), ActivBiotics Web site (January 2008), Google Scholar, and searches of selected bibliographies using the terms rifamycin, ansamycins, rifampin, rifabutin, rifampicin, rifaximin, rifalazil, Clostridium difficile, C. difficile, and CDAD. STUDY SELECTION AND DATA EXTRACTION: In vivo and in vitro studies investigating the use of rifamycins for CDAD were selected, along with all clinical trials using rifamycins in patients with CDAD. DATA SYNTHESIS: Nine studies totaling 890 isolates were identified that investigated the in vitro susceptibility of rifampin (6 studies), rifaximin (3 studies), and rifalazil (2 studies). Rifamycins consistently displayed potent activity against tested strains, although strains with decreased susceptibility have been identified. Six published clinical studies involving 81 patients have investigated the use of rifamycins for the treatment of CDAD. These have generally been small studies, although initial positive clinical results have been reported on the use of rifamycins for recurrent CDAD. CONCLUSIONS: Preliminary data support the use of rifamycins for treatment of CDAD. With the increased incidence and severity of CDAD, further investigation into this drug class as a treatment regimen for CDAD is warranted. PMID- 18430793 TI - Effect of an academic detailing intervention on the utilization rate of cyclooxygenase-2 inhibitors in the elderly. AB - BACKGROUND: Osteoarthritis is prevalent in the elderly. Nova Scotia general practitioners (GPs) identified the need for an academic detailing (AD) intervention aimed at optimizing the management of osteoarthritis. AD was provided by Dalhousie University Continuing Medical Education in a face-to-face encounter employing evidence-based information. GP participation was voluntary. OBJECTIVE: To evaluate the effect of a GP-targeted osteoarthritis AD intervention on a reduction in the prescribing of cyclooxygenase-2 (COX-2) inhibitors, as well as examine the intervention effect on the utilization rates of gastroprotective agents and medical services. METHODS: A retrospective cohort study design employing administrative data was used. Differences in utilization rates between intervention and control groups were evaluated using generalized estimating equations analysis for longitudinal data over four 90-day postintervention periods. Confounding was addressed using propensity scores to adjust for between group bias on the measured covariates. RESULTS: The between-group difference for change in COX-2 utilization rates was 0.76 defined daily doses/patient (p = 0.040; 95% CI 0.037 to 1.48) for the 3-month period following the intervention, with lower COX-2 utilization in the AD intervention group than in the control group. The intervention group showed a significant decrease in the within-group utilization rate between the pre- and postintervention periods (z =-2.34; p = 0.019). The between-group difference for change in GP office visit rates was 0.40 visits/patient (p = 0.028; 95% CI 0.046 to 0.79) with the intervention group, showing higher visit rates compared with the control group. CONCLUSIONS: The osteoarthritis AD intervention was associated with a significant decrease (23%) in COX-2 utilization rates in the 3-month period immediately following the intervention. The only secondary outcome to show a significant between-group effect was the GP office visit rate, which was higher for the intervention group in the second 3-month postintervention period. PMID- 18430794 TI - Mesalamine-associated hypersensitivity myocarditis in ulcerative colitis. PMID- 18430795 TI - Identification of interleukin-13 receptor alpha2 chain overexpression in situ in high-grade diffusely infiltrative pediatric brainstem glioma. AB - Human malignant glioma cell lines and adult brain tumors overexpress high levels of interleukin-13 receptor alpha2 chain (IL-13Ralpha2). Because the IL-13Ralpha2 chain is an important target for cancer therapy and prognosis for patients with brainstem glioma (BSG) remains dismal, we investigated the expression of this receptor in specimens of diffusely infiltrative pediatric BSG relative to normal brain tissue. Twenty-eight BSG specimens and 15 normal brain specimens were investigated for IL-13Ralpha2 protein expression by immunohistochemical analysis (IHC) using two different antibodies in two different laboratories. Highly sensitive Q-dot-based IHC and in situ hybridization (ISH) assays were also developed to identify IL-13Ralpha2 protein and RNA in these specimens. The results were evaluated independently in two laboratories in a blinded fashion. By Q-dot IHC or a standard IHC assay, 17 of 28 (61%) tumor specimens showed modest to strong staining for IL-13Ralpha2, while 15 normal brain tissue samples showed weak expression for IL-13Ralpha2 protein. Significant interrater agreement between the two laboratories was seen in the assessment of IL-13Ralpha2 intensity. High-level IL-13Ralpha2 RNA expression was detected in tumor samples by Q-dot ISH, but only weak RNA expression was observed in normal brain. Significant agreement between ISH and IHC assays was observed (simple kappa [kappa] estimate=0.358, weighted kappa=0.89, p=0.001). IL-13Ralpha2 protein and mRNA are expressed to significantly higher levels in BSG than in normal brain tissue. Both IHC and ISH represent robust methods to detect expression of the IL 13Ralpha2 receptor in BSG that could represent an important new drug target for treatment of this disease. PMID- 18430796 TI - FDG-PET imaging for the evaluation of antiglioma agents in a rat model. AB - The increasing development of novel anticancer agents demands parallel advances in the methods used to rapidly assess their therapeutic efficacy (TE) in the preclinical phase. We evaluated the ability of small-animal PET, using the (18)F fluoro-deoxy-D-glucose (FDG) radiotracer, to predict the TE of a number of anticancer agents in the rat C6 glioma model following 3 days of treatment. Semi quantitative measurements of changes in FDG uptake during the course of treatment (standardized uptake value response [SUV(r)]) were found to be significantly lower in tumors treated with the hypoxia-inducible factor-1alpha inhibitor YC-1 (15 mg/kg) than in tumors in the control group. No significant SUV(r) change was observed following a similar 3-day regimen with the proapoptotic agent NS1619 (20 microg/kg), the combination of YC-1 and NS1619, or the alkylating agent temozolomide (7.5 mg/kg). Quantitative immunohistochemical studies demonstrated significantly lower levels of glucose transporter-1 (GLUT-1) expression in the YC 1-treated tumors, thereby correlating with the low SUV(r) observed in this group. The ability of SUV(r) to predict gold-standard outcomes of TE was further validated as YC-1-treated tumors had decreased volumes compared to control tumors. As such, we successfully demonstrated the ability of FDG-PET to rapidly determine the TE of novel agents for the treatment of glioma in the preclinical phase of evaluation. PMID- 18430797 TI - Selective delivery of beta cell antigen to dendritic cells in vivo leads to deletion and tolerance of autoreactive CD8+ T cells in NOD mice. AB - Type 1 diabetes (T1D) is an autoimmune disease resulting from defects in central and peripheral tolerance and characterized by T cell-mediated destruction of islet beta cells. Cytotoxic CD8(+) T cells, reactive to beta cell antigens, are required for T1D development in the NOD mouse model of the disease, and CD8(+) T cells specific for beta cell antigens can be detected in the peripheral blood of T1D patients. It has been evident that in nonautoimmune-prone mice, dendritic cells (DCs) present model antigens in a tolerogenic manner in the steady state, e.g., in the absence of infection, and cause T cells to proliferate initially but then to be deleted or rendered unresponsive. However, this fundamental concept has not been evaluated in the setting of a spontaneous autoimmune disease. To do so, we delivered a mimotope peptide, recognized by the diabetogenic CD8(+) T cell clone AI4, to DCs in NOD mice via the endocytic receptor DEC-205. Proliferation of transferred antigen-specific T cells was initially observed, but this was followed by deletion. Tolerance was achieved because rechallenge of mice with the mimotope peptide in adjuvant did not induce an immune response. Thus, targeting of DCs with beta cell antigens leads to deletion of autoreactive CD8(+) T cells even in the context of ongoing autoimmunity in NOD mice with known tolerance defects. Our results provide support for the development of DC targeting of self antigens for treatment of chronic T cell-mediated autoimmune diseases. PMID- 18430798 TI - Loss of Hoxb8 alters spinal dorsal laminae and sensory responses in mice. AB - Although Hox gene expression has been linked to motoneuron identity, a role of these genes in development of the spinal sensory system remained undocumented. Hoxb genes are expressed at high levels in the dorsal horn of the spinal cord. Hoxb8 null mutants manifest a striking phenotype of excessive grooming and hairless lesions on the lower back. Applying local anesthesia underneath the hairless skin suppressed excessive grooming, indicating that this behavior depends on peripheral nerve activity. Functional ablation of mouse Hoxb8 also leads to attenuated response to nociceptive and thermal stimuli. Although spinal ganglia were normal, a lower postmitotic neural count was found in the dorsalmost laminae at lumbar levels around birth, leading to a smaller dorsal horn and a correspondingly narrowed projection field of nociceptive and thermoceptive afferents. The distribution of the dorsal neuronal cell types that we assayed, including neurons expressing the itch-specific gastrin-releasing peptide receptor, was disorganized in the lumbar region of the mutant. BrdU labeling experiments and gene-expression studies at stages around the birth of these neurons suggest that loss of Hoxb8 starts impairing development of the upper laminae of the lumbar spinal cord at approximately embryonic day (E)15.5. Because none of the neuronal markers used was unexpressed in the adult dorsal horn, absence of Hoxb8 does not impair neuronal differentiation. The data therefore suggest that a lower number of neurons in the upper spinal laminae and neuronal disorganization in the dorsal horn underlie the sensory defects including the excessive grooming of the Hoxb8 mutant. PMID- 18430799 TI - Glutamic acid 242 is a valve in the proton pump of cytochrome c oxidase. AB - Aerobic life is based on a molecular machinery that utilizes oxygen as a terminal electron sink. The membrane-bound cytochrome c oxidase (CcO) catalyzes the reduction of oxygen to water in mitochondria and many bacteria. The energy released in this reaction is conserved by pumping protons across the mitochondrial or bacterial membrane, creating an electrochemical proton gradient that drives production of ATP. A crucial question is how the protons pumped by CcO are prevented from flowing backwards during the process. Here, we show by molecular dynamics simulations that the conserved glutamic acid 242 near the active site of CcO undergoes a protonation state-dependent conformational change, which provides a valve in the pumping mechanism. The valve ensures that at any point in time, the proton pathway across the membrane is effectively discontinuous, thereby preventing thermodynamically favorable proton back-leakage while maintaining an overall high efficiency of proton translocation. Suppression of proton leakage is particularly important in mitochondria under physiological conditions, where production of ATP takes place in the presence of a high electrochemical proton gradient. PMID- 18430800 TI - Complex genetic interactions underlying expression differences between Drosophila races: analysis of chromosome substitutions. AB - Regulation of gene expression is usually separated into cis and trans components. The separation may become artificial if much of the variation in expression is under multigenic and epistatic (e.g., cis-by-trans) control. There is hence a need to quantify the relative contribution of cis, trans, and cis-by-trans effects on expression divergence at different levels of evolution. To do so across the whole genome, we analyzed the full set of chromosome-substitution lines between the two behavioral races of Drosophila melanogaster. Our observations: (i) Only approximately 3% of the genes with an expression difference are purely cis regulated. In fact, relatively few genes are governed by simple genetics because nearly 80% of expression differences are controlled by at least two chromosomes. (ii) For 14% of the genes, cis regulation does play a role but usually in conjunction with trans regulation. This joint action of cis and trans effects, either additive or epistatic, is referred to as inclusive cis effect. (iii) The percentage of genes with inclusive cis effect increases to 32% among genes that are strongly differentiated between the two races. (iv) We observed a nonrandom distribution of trans-acting factors, with a substantial deficit on the second chromosome. Between Drosophila racial groups, trans regulation of expression difference is extensive, and cis regulation often evolves in conjunction with trans effects. PMID- 18430801 TI - Role of epsin 1 in synaptic vesicle endocytosis. AB - Epsin has been suggested to act as an alternate adaptor in several endocytic pathways. Its role in synaptic vesicle recycling remains, however, unclear. Here, we examined the role of epsin in this process by using the lamprey reticulospinal synapse as a model system. We characterized a lamprey ortholog of epsin 1 and showed that it is accumulated at release sites at rest and also at clathrin coated pits in the periactive zone during synaptic activity. Disruption of epsin interactions, by presynaptic microinjection of antibodies to either the epsin-N terminal homology domain (ENTH) or the clathrin/AP2 binding region (CLAP), caused profound loss of vesicles in stimulated synapses. CLAP antibody-injected synapses displayed a massive accumulation of distorted coated structures, including coated vacuoles, whereas in synapses perturbed with ENTH antibodies, very few coated structures were found. In both cases coated pits on the plasma membrane showed a shift to early intermediates (shallow coated pits) and an increase in size. Moreover, in CLAP antibody-injected synapses flat clathrin-coated patches occurred on the plasma membrane. We conclude that epsin is involved in clathrin mediated synaptic vesicle endocytosis. Our results support a model, based on in vitro studies, suggesting that epsin coordinates curvature generation with coat assembly and further indicating that epsin limits clathrin coat assembly to the size of newly formed vesicles. We propose that these functions of epsin 1 provide an additional mechanism for generation of uniformly sized synaptic vesicles. PMID- 18430802 TI - Renin inhibition and atherosclerosis. PMID- 18430803 TI - Analysis of cortical arrays from Tradescantia virginiana at high resolution reveals discrete microtubule subpopulations and demonstrates that confocal images of arrays can be misleading. AB - Cortical microtubule arrays are highly organized networks involved in directing cellulose microfibril deposition within the cell wall. Their organization results from complex interactions between individual microtubules and microtubule associated proteins. The precise details of these interactions are often not evident using optical microscopy. Using high-resolution scanning electron microscopy, we analyzed extensive regions of cortical arrays and identified two spatially discrete microtubule subpopulations that exhibited different stabilities. Microtubules that lay adjacent to the plasma membrane were often bundled and more stable than the randomly aligned, discordant microtubules that lay deeper in the cytoplasm. Immunolabeling revealed katanin at microtubule ends, on curves, or at sites along microtubules in line with neighboring microtubule ends. End binding 1 protein also localized along microtubules, at microtubule ends or junctions between microtubules, and on the plasma membrane in direct line with microtubule ends. We show fine bands in vivo that traverse and may encircle microtubules. Comparing confocal and electron microscope images of fluorescently tagged arrays, we demonstrate that optical images are misleading, highlighting the fundamental importance of studying cortical microtubule arrays at high resolution. PMID- 18430804 TI - The coi1-16 mutant harbors a second site mutation rendering PEN2 nonfunctional. PMID- 18430805 TI - Genomic analysis reveals poor separation of human cardiomyopathies of ischemic and nonischemic etiologies. AB - Clinically, the differentiation between ischemic (ICM) and nonischemic (NICM) human cardiomyopathies is highly relevant, because ICM and NICM differ with respect to prognosis and certain aspects of pharmacological therapy, despite a common final phenotype characterized by ventricular dilatation and reduced contractility. So far, it is unclear whether microarray-based signatures can be used to infer the etiology of heart failure. Using three different classification algorithms, we independently analyzed one cDNA and two publicly available high density oligonucleotide microarray studies comprising a total of 279 end-stage human heart failure samples. When classifiers identified in a single study were applied to the remaining studies, misclassification rates >25% for ICM and NICM specimens were noted, indicating poor separation of both etiologies. However, data mining of 458 classifier genes that were concordantly identified in at least two of the three data sets points to different biological processes in ICM vs. NICM. Consistent with the underlying ischemia, cytokine signaling pathways and immediate-early response genes were overrepresented in ICM samples, whereas NICM samples displayed a deregulation of cytoskeletal transcripts, genes encoding for the major histocompatibility complex, and antigen processing and presentation pathways, potentially pointing to immunologic processes in NICM. Overall, our results suggest that ICM and NICM exhibit substantial heterogeneity at the transcriptomic level. Prospective studies are required to test whether etiology specific gene expression patterns are present at earlier disease stages or in subsets of both etiologies. PMID- 18430806 TI - Transcriptional profile of right ventricular tissue during acute pulmonary embolism in rats. AB - Acute pulmonary embolism (PE) is the third leading cause of cardiovascular death in the United States. Moderate to severe PE can cause pulmonary arterial hypertension (PH) with resultant right ventricular (RV) heart damage. The mechanisms leading to RV failure after PE are not well defined, although it is becoming clear that PH-induced inflammatory responses are involved. We previously demonstrated profound neutrophil-mediated inflammation and RV dysfunction during PE that was associated with increased expression of several chemokine genes. However, a complete assessment of transcriptional changes in RVs during PE is still lacking. We have now used DNA microarrays to assess the alterations in gene expression in RV tissue during acute PE/PH in rats. Key results were confirmed with real-time RT-PCR. Nine CC-chemokine genes (CCL-2, -3, -4, -6, -7, -9, -17, 20, -27), five CXC-chemokine genes (CXCL-1, -2, -9, -10, -16), and the receptors CCR1 and CXCR4 were upregulated after 18 h of moderate PE, while one C-chemokine (XCL-1) and one CXC-chemokine (CXCL-12) were downregulated. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses indicated increased expression of many inflammatory genes. There was also a major shift in the expression of components of metabolic pathways, including downregulation of fatty acid transporters and oxidative enzymes, a change in glucose transporters, and upregulation of stretch-sensing and hypoxia-inducible transcription factors. This pattern suggests an extensive shift in cardiac physiology favoring the expression of the "fetal gene program." PMID- 18430807 TI - Physiological and molecular evidence of heat acclimation memory: a lesson from thermal responses and ischemic cross-tolerance in the heart. AB - Sporadic findings in humans suggest that reinduction of heat acclimation (AC) after its loss occurs markedly faster than that during the initial AC session. Animal studies substantiated that the underlying acclimatory processes are molecular. Here we test the hypothesis that faster reinduction of AC (ReAC) implicates "molecular memory." In vivo measurements of colonic temperature profiles during heat stress and ex vivo assessment of cross-tolerance to ischemia reperfusion or anoxia insults in the heart demonstrated that ReAC only needs 2 days vs. the 30 days required for the initial development of AC. Stress gene profiling in the experimental groups highlighted clusters of transcriptionally activated genes (37%), which included heat shock protein (HSP) genes, antiapoptotic genes, and chromatin remodeling genes. Despite a return of the physiological phenotype to its preacclimation state, after a 1 mo deacclimation (DeAC) period, the gene transcripts did not resume their preacclimation levels, suggesting a dichotomy between genotype and phenotype in this system. Individual detection of hsp70 and hsf1 transcripts agreed with these findings. HSP72, HSF1/P HSF1, and Bcl-xL protein profiles followed the observed dichotomized genomic response. In contrast, HSP90, an essential cytoprotective component mismatched transcriptional activation upon DeAC. The uniform activation of the similarly responding gene clusters upon De-/ReAC implies that reacclimatory phenotypic plasticity is associated with upstream denominators. During AC, DeAC, and ReAC, the maintenance of elevated/phosphorylated HSF1 protein levels and transcriptionally active chromatin remodeling genes implies that chromatin remodeling plays a pivotal role in the transcriptome profile and in preconditioning to rapid cytoprotective acclimatory memory. PMID- 18430808 TI - Alternative splicing and exon duplication generates 10 unique porcine 5-HT 4 receptor splice variants including a functional homofusion variant. AB - 5-HT(4) receptors are present in human and porcine atrial myocytes while they are absent from the hearts of small laboratory animals. The pig is therefore the only available nonprimate animal model in which to study cardiac 5-HT(4) receptor function under physiological conditions. While several human splice variants of the 5-HT(4) receptor have been described, the splicing behavior of this receptor in porcine tissue is currently unknown. Here we report on the identification of nine novel COOH-terminal splice variants of the porcine 5-HT(4) receptor, which were named 5-HT(4(b2, j, k, l, m, o, p, q, r)). The internal h-variant was found in combination with several COOH-terminal exons. In addition, splice variants were found that comprised duplicated exons fused to the common region of the 5 HT(4) receptor, thereby providing evidence for a duplication of the porcine HTR4 gene. One of these variants putatively encoded a nine transmembrane-spanning domain homofusion receptor, 5-HT(4(9TM)); also the other variants with a duplicated region might translate into functional, transcriptionally fused dimeric 5-HT(4) receptor variants. The elucidation of the genomic context confirmed that the variants were not genomic artefacts but originated from alternative splicing. This was further corroborated by a functional analysis of the variants 5-HT(4(a)), 5-HT(4(r)), and 5-HT(4(9TM)). To our knowledge, our data are the first to report on a functional GPCR with more than seven predicted transmembrane domains. These findings urge for caution when interpreting data on 5-HT(4) receptor-related pharmacology obtained in the pig; validation at the molecular level might be needed before extrapolating results to human. PMID- 18430809 TI - Molecular networks in Dahl salt-sensitive hypertension based on transcriptome analysis of a panel of consomic rats. AB - The Dahl salt-sensitive (SS) rat is a widely used model of human salt-sensitive hypertension and renal injury. We studied the molecular networks that underlie the complex disease phenotypes in the SS model, using a design that involved two consomic rat strains that were protected from salt-induced hypertension and one that was not protected. Substitution of Brown Norway (BN) chromosome 13 or 18, but not 20, into the SS genome was found to significantly attenuate salt-induced hypertension and albuminuria. Gene expression profiles were examined in the kidneys of SS and consomic SS-13(BN), SS-18(BN), and SS-20(BN) rats with a total of 240 cDNA microarrays. The substituted chromosome was overrepresented in genes differentially expressed between a consomic strain and SS rats on a 0.4% salt diet. F5, Serpinc1, Slc19a2, and genes represented by three other expressed sequence tags (ESTs), which are located on chromosome 13, were found to be differentially expressed between SS-13(BN) and all other strains examined. Likewise, Acaa2, B4galt6, Colec12, Hsd17b4, and five other ESTs located on chromosome 18 exhibited expression patterns unique to SS-18(BN). On exposure to a 4% salt diet, there were 184 ESTs in the renal cortex and 346 in the renal medulla for which SS-13(BN) and SS-18(BN) shared one expression pattern, while SS and SS-20(BN) shared another, mirroring the phenotypic segregation among the four strains. Molecular networks that might contribute to the development of Dahl salt sensitive hypertension and albuminuria were constructed with an approach that merged biological knowledge-driven analysis and data-driven Bayesian probabilistic analysis. PMID- 18430810 TI - Functional meta-analysis of double connectivity in gene coexpression networks in mammals. AB - In functional genomics, the high-throughput methods such as microarrays 1) allow analysis of the relationships between genes considering them as elements of a network and 2) lead to biological interpretations thanks to Gene Ontology. But up to now it has not been possible to find relationships between the functions and the connectivity of the genes in coexpression networks. To achieve this aim, we have defined a double connectivity for each gene by the numbers of its significant negative and positive correlations with the other genes within a given biological condition, or group. Here, based on the analysis of 1,260 DNA microarrays, we show that this double connectivity clearly separates two types of genes, those with a predominantly strong negative connectivity, hub- genes, and those with a predominantly strong positive connectivity, hub+ genes. Interestingly, the hub+ genes concerned transcription factors more often than by chance and, similarly, for the hub- genes concerning miRNA predicted targets. Furthermore, a meta-analysis of GO annotations carried out on 67 groups in humans and rats shows that these two types of genes correspond to a functional biological duality. The hub- genes were mainly involved in basic functions common to all eukaryote cells, whereas the hub+ genes were mainly involved in specialized functions related to cell differentiation and communication. The separation and the biological role of these hub- and hub+ genes provide a powerful new tool for a better understanding of the control and regulation of the key genes involved in cellular differentiation and physiopathological conditions. PMID- 18430811 TI - Abstracts of Posters Presented at the 40th Annual Meeting of the Society for Obstetric Anesthesia and Perinatology. Chicago, Illinois, USA. April 30-May 4, 2008. PMID- 18430812 TI - Bilateral hydrosalpinx in adolescent girls with Hirschsprung's disease: association of two rare conditions. AB - OBJECTIVE: The purpose of our study was to illustrate three cases of bilateral hydrosalpinx in postpubertal girls operated on for Hirschsprung's disease and to discuss the possible cause: iatrogenic or congenital. We identified bilateral hydrosalpinx in three postpubertal sexually inactive girls with Hirschsprung's disease treated, respectively by Duhamel, Soave pull-through, and Martin procedures. No history of surgical complications or pelvic inflammation had been reported. CONCLUSION: Hirschsprung's disease is rare in girls and bilateral hydrosalpinx is also extremely uncommon in sexually inactive adolescents. We think there may be a possible common cause: either a postsurgical complication or a congenital defect of the autonomous innervation in the context of a neurocristopathy. Because of the rarity of both conditions, the association is unlikely to be coincidental. The cause of this association is unclear and further studies are required to find its prevalence and to estimate the possible impact on fertility. PMID- 18430813 TI - CT-guided biopsy of bone: a radiologist's perspective. AB - OBJECTIVE: We present an overview of approaches for bone biopsy used to minimize potential tumor seeding of adjacent soft-tissue structures and compartments. We discuss a variety of approaches related to specific anatomic parts and review pertinent anatomy. CONCLUSION: We provide important guidelines and key examples that will help readers perform percutaneous needle bone biopsy safely. PMID- 18430814 TI - MRI of hepatic adenomatosis: initial observations with gadoxetic acid contrast agent in three patients. AB - OBJECTIVE: The purpose of our study was to describe the MR signal characteristics of histologically proven liver adenomatosis in three patients using gadoxetic acid, a newly developed liver-specific MR contrast agent. CONCLUSION: In all three patients, more than 100 liver adenomas revealed no metabolism of the new liver-specific contrast agent in the delayed phase. Because of absent or strongly reduced intracellular uptake of gadoxetic acid in all adenomas during delayed contrast-enhanced series, differentiation of adenomas from dysplastic or malignant lesions was not possible. PMID- 18430815 TI - Cardiac valve disease: spectrum of findings on cardiac 64-MDCT. AB - OBJECTIVE: Recent studies have established that cardiac MDCT generates high quality images of the cardiac valves. Images are acquired during a single breath hold (inspiration) after the injection of iodinated contrast material (5 mL/s) followed by a saline bolus chaser. Incremental data sets are then reconstructed throughout the R-R interval, and after transfer to a workstation, specialized software combines data sets sequentially to generate cine loops of the heart throughout the cardiac cycle. The purpose of this article is to describe the cardiac MDCT techniques allowing optimal cardiac valve depiction and to illustrate the MDCT appearances of the most important valve diseases. CONCLUSION: Cardiac MDCT provides an excellent imaging method for illustrating cardiac valve disease. Radiologists should be aware of the various appearances of the common and most important cardiac valve diseases on cardiac MDCT. PMID- 18430816 TI - Neuroimaging of tuberous sclerosis: spectrum of pathologic findings and frontiers in imaging. AB - OBJECTIVE: The purpose of this article is to present neuroimaging findings in tuberous sclerosis complex, including recently developed imaging techniques that have demonstrated clinical benefit to this patient population. CONCLUSION: Neuroimaging advances have improved the diagnosis of tuberous sclerosis complex and the treatment of children with this condition. Superimposition of functional information from PET onto MRI allows accurate and noninvasive identification of epileptogenic tubers, improving surgical cure rates. Magnetic source imaging can also be used to localize epileptiform activity arising from tubers. PMID- 18430817 TI - Midterm results of percutaneous CT-guided aspiration of symptomatic lumbar discal cysts. AB - OBJECTIVE: Lumbar intervertebral discal cysts are rarely reported. To our knowledge, only 12 cases have been reported in the literature. The symptoms are indistinguishable from those of typical discal herniation. Our aim is to report the imaging characteristics, method of percutaneous CT-guided aspiration, and clinical outcome after management of lumbar intervertebral discal cysts. CONCLUSION: Percutaneous CT-guided aspiration is an effective method for the management of lumbar intervertebral discal cysts. PMID- 18430818 TI - Bronchopleural cutaneous fistula. PMID- 18430819 TI - Adverse effects of intratendinous injection. PMID- 18430820 TI - Superficial thinking. PMID- 18430821 TI - The diagnostic radiology exam of the future: the law of unintended consequences meets the law of supply and demand. PMID- 18430822 TI - My old Kentucky home, goodnight: potential impact of planned changes in the radiology board certification process. PMID- 18430823 TI - "CT of the solitary pulmonary nodule"--a commentary. PMID- 18430824 TI - Gestalt theory: implications for radiology education. AB - OBJECTIVE: The Gestalt theory of modern psychology is grounded in the ideas that holistic rather than atomistic approaches are necessary to understand the mind, and that the mental whole is greater than the sum of its component parts. Although the Gestalt school fell out of favor due to its descriptive rather than explanatory nature, it permanently changed our understanding of perception. For the radiologist, such fundamental Gestalt concepts as figure-ground relationships and a variety of "grouping principles" (the laws of closure, proximity, similarity, common region, continuity, and symmetry) are ubiquitous in daily work, not to mention in art and personal life. CONCLUSION: By considering the applications of these principles and the stereotypical ways in which humans perceive visual stimuli, a radiology learner may incur fewer errors of diagnosis. This article serves to introduce several important principles of Gestalt theory, identify examples of these principles in widely recognizable fine art, and highlight their implications for radiology education. PMID- 18430825 TI - Teleradiology coming of age: winners and losers. PMID- 18430826 TI - The incidental adrenal mass on CT: prevalence of adrenal disease in 1,049 consecutive adrenal masses in patients with no known malignancy. AB - OBJECTIVE: The purpose of our study was to determine the nature and prevalence of adrenal lesions identified on CT in patients with no known malignancy. MATERIALS AND METHODS: A computer search of abdominal CT reports using the term "adrenal" was performed in 65,231 consecutive patients with examinations performed from January 2000 to December 2003. An adrenal mass was identified in 3,307 (5%) patients. Patients with no known malignancy and no suspicion for a hyperfunctioning adrenal mass were further isolated. Nine hundred seventy-three patients with 1,049 adrenal masses fulfilled the study criteria. The nature of each lesion was determined by histopathology; imaging characterization with CT, MRI, or washout; a minimum of 1 year of stability on follow-up imaging; or clinical follow-up of at least 2 years. RESULTS: One thousand forty-nine adrenal masses were characterized with the following methods: histopathology (n = 12), imaging characterization (n = 909), imaging follow-up (n = 87), and clinical follow-up (n = 41). There were 788 adenomas constituting 75% of all lesions. There were 68 myelolipomas (6%), 47 hematomas (4%), and 13 cysts (1%). Three pheochromocytomas (0.3%) and one cortisol-producing adenoma (0.1%) were found incidentally. One hundred twenty-eight lesions (12%) were presumed to be benign by imaging or clinical stability. No malignant adrenal masses were found, even among the 14 patients who later developed malignancy elsewhere. CONCLUSION: In 973 consecutive patients with an incidental adrenal mass and no history of cancer, no malignant lesions were identified. Adenomas (75%) and myelolipomas (6%) were the most common lesions. PMID- 18430827 TI - Dual-energy CT iodine-subtraction virtual unenhanced technique to detect urinary stones in an iodine-filled collecting system: a phantom study. AB - OBJECTIVE: The objective of our study was to evaluate the feasibility of virtual unenhanced images reconstructed from a dual-energy CT scan to depict urinary stones in an iodine solution in a phantom study. MATERIALS AND METHODS: Twenty urinary stones of different sizes (1.4-4.2 mm in short-axis diameter) were placed in plastic containers. The containers were consecutively filled with different concentrations of iodine solution (21, 43, 64, 85, and 107 mg/dL; CT attenuation value range, 510-2,310 H at 120 kVp). Dual-energy CT was repeated with 80-140 and 100-140 kVp pairs, two collimation-slice thickness combinations, and the presence or absence of a 4-cm-thick oil gel around the phantom. The iodine-subtraction virtual unenhanced images were reconstructed using commercial software. The images were evaluated by three radiologists in consensus for the visibility of the stones and the presence of residual nonsubtracted iodine. Stone visibility rates were compared between the 80-140 and 100-140 kVp pairs and the five different iodine concentrations. RESULTS: Stone visibility rates with the 80-140 kVp pair were 99%, 93%, 96%, 94%, and 3% and those with the 100-140 kVp pair were 98%, 95%, 99%, 94%, and 99% for an iodine concentration of 21, 43, 64, 85, and 107 mg/dL, respectively. The poor visibility rate with 80-140 kVp and 107 mg/dL iodine concentration was due to the failure of iodine subtraction. CONCLUSION: Dual-energy CT iodine-subtraction virtual unenhanced technique is capable of depicting urinary stones in iodine solutions of a diverse range of concentrations in a phantom study. PMID- 18430828 TI - Development of renal scars on CT after abdominal trauma: does grade of injury matter? AB - OBJECTIVE: The objective of our study was to determine whether there is an association between the grade of a traumatic renal injury and the subsequent development of renal parenchymal scars on CT. MATERIALS AND METHODS: We performed a retrospective study encompassing all acute trauma patients admitted to our institution over a 42-month period found to have renal parenchyma injuries on initial MDCT and also to have undergone a follow-up CT performed at least 1 month after trauma. We identified 54 patients who sustained blunt (n = 44) or penetrating (n = 10) abdominal trauma. The renal injuries were graded by two radiologists according to the Organ Injury Scaling Committee of the American Association for the Surgery of Trauma (AAST), grades I through V. Follow-up CT was reviewed for the presence of parenchymal distortion, scarring, or perfusion defects. RESULTS: Of the 54 patients, 12 had grade I injury, eight had grade II injury, 22 had grade III injury, 10 had grade IV injury, and two had grade V injury. Grades I and II traumatic renal injuries were undetectable on follow-up CT. Grade III injuries resulted in the development of renal scars in 14 of 22 (64%) patients. Scarring resulted in all patients with grades IV and V injuries. CONCLUSION: Grades I and II renal injuries heal completely, whereas higher grades of renal trauma result in permanent parenchymal scarring. Hence, incidentally discovered renal scars in patients with a history of minor renal trauma should be attributed tentatively to other causes that may or may not require additional investigation. PMID- 18430829 TI - MRI techniques for prediction of local tumor progression after high-intensity focused ultrasonic ablation of prostate cancer. AB - OBJECTIVE: The purpose of this study was to evaluate the diagnostic performance of dynamic contrast-enhanced MRI (DCE-MRI) and of T2-weighted MRI with diffusion weighted imaging (DWI) for predicting local tumor progression after high intensity focused ultrasonic ablation of localized prostate cancer. MATERIALS AND METHODS: Twenty-seven patients who had increased levels of prostate-specific antigen after high-intensity focused ultrasonic ablation underwent MRI and endorectal biopsy. The MR images and biopsy results were correlated for six prostate sectors. Residual or recurrent prostate cancer after treatment was defined as local tumor progression if the biopsy results showed cancer foci. Two readers blinded to the clinical findings and biopsy results used a 5-point scale to independently assess DCE-MR images and T2-weighted and diffusion-weighted MR images. The results were compared by use of the McNemar test with Bonferroni correction, generalized estimating equations, and receiver operating characteristic analysis. RESULTS: After high-intensity focused ultrasonic ablation, local tumor progression was pathologically detected in 54 (33%) of 162 sectors in 18 patients. The sensitivities of DCE-MRI and T2-weighted MRI with DWI were 80% and 63% for reader 1 (p = 0.004) and 87% and 70% for reader 2 (p = 0.004). The specificities of DCE-MRI and T2-weighted MRI with DWI were 68% and 78% for reader 1 (p = 0.002) and 63% and 74% for reader 2 (p < 0.001). The accuracy rates of DCE-MRI and T2-weighted MRI with DWI were 72% and 73% for reader 1 (p > 0.05) and 71% and 73% for reader 2 (p > 0.05). The areas under the receiver operating characteristic curve for DCE-MRI and T2-weighted MRI with DWI were 0.77 and 0.77 for reader 1 and 0.85 and 0.81 for reader 2. CONCLUSION: For prediction of local tumor progression of prostate cancer after high-intensity focused ultrasonic ablation, DCE-MRI was more sensitive than T2-weighted MRI with DWI, but T2-weighted MRI with DWI was more specific than DCE-MRI. PMID- 18430830 TI - Endorectal and dynamic contrast-enhanced MRI for detection of local recurrence after radical prostatectomy. AB - OBJECTIVE: The objective of our study was to evaluate the sensitivity and specificity of endorectal MRI combined with dynamic contrast-enhanced MRI to detect local recurrence after radical prostatectomy. MATERIALS AND METHODS: A total of 51 patients who had undergone radical prostatectomy for prostatic adenocarcinoma 10 months to 6 years before underwent a combined endorectal coil MRI and dynamic gadolinium-enhanced MRI before endorectal sonographically guided biopsy of the prostatic fossa. The MRI combined with MR dynamic imaging results were correlated with the presence of recurrence defined as a positive biopsy result or reduction in prostate-specific antigen level after radiation therapy. RESULTS: Overall data of 46 (25 recurred, 21 nonrecurred) out of 51 evaluated patients were analyzed. All recurrences showed signal enhancement after gadolinium administration and, in particular, 22 of 24 patients (91%) showed rapid and early signal enhancement. The overall sensitivity and specificity of MR dynamic imaging was higher compared with MRI alone (88%, [95% CI] 69-98% and 100%, 84-100% compared with 48%, 28-69% and 52%, 30-74%). MRI combined with dynamic imaging allowed better identification of recurrences compared with MRI alone (McNemar test: chi-square(1) = 16.67; p = < 0.0001). CONCLUSION: MRI combined with dynamic contrast-enhanced MRI showed a higher sensitivity and specificity compared with MRI alone in detecting local recurrences after radical prostatectomy. PMID- 18430831 TI - Symptomatic simple renal cyst: comparison of continuous negative-pressure catheter drainage and single-session alcohol sclerotherapy. AB - OBJECTIVE: The purpose of this study was to evaluate whether continuous percutaneous catheter drainage with negative pressure yields better results than single-session alcohol sclerotherapy in the management of symptomatic simple renal cysts. SUBJECTS AND METHODS: Eighty-five patients with 92 simple renal cysts were randomly assigned to two groups in a prospective controlled trial. One group was treated with sonographically guided continuous catheter drainage with negative pressure and the other group with single-session alcohol sclerotherapy. Patient demographics, clinical characteristics, treatment outcome, and complications were analyzed. RESULTS: The initial volume of the cysts did not differ significantly between the groups, but the final volume was significantly smaller in the continuous drainage group (p = 0.026). During the 24-month follow up period, 37 (40%) of the cysts disappeared completely: 24 (52%) of the 46 cysts in the drainage group and 13 (28%) of the 46 cysts in the sclerotherapy group (p = 0.033). In the sclerotherapy group, the probability of disappearance of the cysts was highly dependent on cyst size, being less for giant cysts (p = 0.01). Cyst size was not a significant factor in probability of disappearance in the drainage group (p = 0.15). The probability of disappearance of giant cysts (volume > 500 mL) differed significantly between the groups (p = 0.009), but there was no difference in probability of disappearance of moderately large cysts (p = 0.16). Three of 14 patients with giant cysts in the drainage group and 10 of 13 such patients in the sclerotherapy group had recurrences that necessitated additional treatment (p = 0.007). They were successfully treated with continuous catheter drainage. CONCLUSION: Continuous catheter drainage with negative pressure is more efficient than single-session alcohol sclerotherapy in the management of giant cysts. For moderately large cysts, the two methods have similar results. PMID- 18430833 TI - Clinical application of the BI-RADS final assessment to breast sonography in conjunction with mammography. AB - OBJECTIVE: The objective of our study was to report the results of classification of sonographic findings according to BI-RADS and to calculate the positive predictive value (PPV) for each BI-RADS assessment category. SUBJECTS AND METHODS: We prospectively classified 4,668 breast sonograms according to BI-RADS final assessment category. Suspicious sonographic findings were divided into major and minor suspicious findings. Category 1 was normal and category 2 was a benign finding such as cyst or nodule with uniform and intense hyperechogenicity. A nodule neither category 2 nor category 4 or 5 was defined as category 3. A nodule with one or more suspicious findings, not category 5, was defined as category 4. A nodule with two or more major suspicious findings was defined as category 5. RESULTS: Of the 4,668 cases, 321 cases failed to undergo follow-up of at least 1 year. The PPV was 0.1% in category 1 (3/2,191), 0% in category 2 (0/773), 0.8% in category 3 (6/737), 31.1% in category 4 (161/519), and 96.9% in category 5 (123/127). In palpable lesions (n = 751), the PPV was 2.2% in category 1 (2/93), 0.9% in category 3 (2/217), 54% in category 4 (107/198), and 98% in category 5 (98/100). In nonpalpable lesions (n = 3,596), the PPV was 0.05% in category 1 (1/2,098), 0.8% in category 3 (4/520), 16.8% in category 4 (54/321), and 92.6% in category 5 (25/27). CONCLUSION: As with mammography, placing sonographic lesions into BI-RADS categories is useful for predicting the presence of malignancy. PMID- 18430832 TI - Accuracy of short-interval follow-up mammograms by patient and radiologist characteristics. AB - OBJECTIVE: The purpose of our study was to examine the accuracy of short-interval follow-up mammograms and evaluate patient and radiologist characteristics associated with accuracy. MATERIALS AND METHODS: We evaluated 45,007 initial short-interval follow-up mammograms from the Breast Cancer Surveillance Consortium interpreted 3-9 months after a probably benign assessment on a screening or diagnostic examination between 1994 and 2004. We linked these mammograms with patient characteristics and breast cancer diagnoses within 12 months. A subset of short-interval follow-up examinations (n = 13,907) was merged with radiologist characteristics collected from survey data from 130 interpreting radiologists. Using logistic regression, we fit generalized estimating equations to model sensitivity and specificity of short-interval follow-up mammograms by patient and radiologist characteristics. RESULTS: For every 1,000 women, 8.0 women (0.8%) were diagnosed with breast cancer within 6 months and 11.3 (1.1%) within 12 months. Sensitivity was 83.3% (95% CI, 79.4-87.3%) for cancers diagnosed within 6 months and 60.5% (56.2-64.7%) for those diagnosed within 12 months. Specificity was 97.2% (96.9-97.6%) at 6 months and 97.3% (96.9-97.6%) at 12 months. Sensitivity at 12 months increased among women with unilateral short interval follow-up mammograms (odds ratio, 1.56 [95% CI, 1.06-2.29]) and when the interpreting radiologist spent more than 10 hours a week in breast imaging (odds ratio, 3.25 [1.00-10.52]). CONCLUSION: Initial short-interval follow-up mammography examinations had a lower sensitivity for detecting breast cancer within 12 months than other diagnostic mammograms (61% for short-interval follow up vs 80% for diagnostic mammograms reported in the literature). However, sensitivity within the 6-month interval that is usually recommended for subsequent follow-up was 83%. Accuracy of short-interval follow-up mammograms was influenced by few patient and radiologist characteristics. PMID- 18430834 TI - Predicting neonatal deaths and pulmonary hypoplasia in isolated congenital diaphragmatic hernia using the sonographic fetal lung volume-body weight ratio. AB - OBJECTIVE: The objective of our study was to evaluate the potential of the sonographic fetal lung volume-body weight ratio to predict neonatal deaths and pulmonary hypoplasia in fetuses with isolated congenital diaphragmatic hernia (CDH). SUBJECTS AND METHODS: Between January 2002 and December 2004, 40 fetuses with isolated CDH and 450 control subjects were prospectively evaluated in two centers. Fetal lung volumes were estimated on 3D sonography using the rotational technique and fetal weight on 2D sonography using the Hadlock equation. The ratio of sonographic fetal lung volume to body weight was calculated in each case and was correlated with neonatal deaths using the Mann-Whitney U test. Accuracies of the ratio in predicting neonatal deaths and pathologic diagnosis of pulmonary hypoplasia were also evaluated. RESULTS: The ratio of sonographic fetal lung volume to body weight is constant throughout gestation, with a mean value of 0.025. The ratio was significantly lower in neonates that died (median, 0.009; range, 0.004-0.021) than in those that survived (median, 0.011; range, 0.008 0.020) (p = 0.018). Pulmonary hypoplasia was suspected prenatally in 34 of 40 (85.0%) fetuses with CDH, in all cases of death (100%), and in seven of nine (77.8%) neonates that survived. At autopsy, pulmonary hypoplasia was diagnosed in 19 cases (86.4%). Accuracies of the ratio in predicting neonatal deaths and pulmonary hypoplasia were 64.5% (20/31) and 86.4% (19/22), respectively. CONCLUSION: The sonographic fetal lung volume-body weight ratio can be used more accurately to diagnose pulmonary hypoplasia than to predict neonatal deaths in fetuses with isolated CDH. Further studies are necessary to show the prevalence of pulmonary hypoplasia in fetuses with isolated CDH and its importance for predicting neonatal deaths. PMID- 18430835 TI - Submucosal fibroids becoming endocavitary following uterine artery embolization: risk assessment by MRI. AB - OBJECTIVE: The purpose of our study was to assess the relationship between the endometrium and submucosal fibroids before and after uterine artery embolization (UAE). MATERIALS AND METHODS: Contrast-enhanced pelvic 1.5-T MRI was performed in 49 women before and after UAE over a 2-year period. Dominant (largest diameter) fibroids in intramural, submucosal, subserosal, pedunculated subserosal, and endocavitary locations were assessed on pre- (baseline) and postembolization MRI. Size, locations of dominant fibroids relative to endometrium and serosa before and after embolization were compared. The ratio between the largest endometrial interface and the maximum dimension of the dominant submucosal fibroid (interface dimension ratio) was determined on baseline MRI. The infarction rate for dominant fibroids was estimated after UAE. RESULTS: One hundred forty dominant fibroids were identified on baseline MRI. Forty-nine (35%) were intramural, 39 (28%) were submucosal, 34 (24%) were subserosal, eight (6%) were pedunculated subserosal, and 10 (6%) were endocavitary in location on preembolization MRI. After UAE, of 39 dominant submucosal fibroids, 13 (33%) became endocavitary: complete (n = 4), partial (n = 9) on the basis of European Society of Gynaecological Endoscopy (ESGE) classification. The preembolization mean interface-dimension ratio and mean diameters for dominant fibroids that became endocavitary were significantly greater than for those that did not become endocavitary after embolization (0.65 vs 0.32, p < 0.005; 8 vs 5.4 cm, p < 0.05, respectively). All dominant submucosal fibroids showed 100% infarction after UAE. CONCLUSION: Submucosal fibroids with an interface-dimension ratio of greater than 0.55 are more likely to migrate into the endometrial cavity after UAE. The majority of these are expelled spontaneously without significant symptoms. Rarely, submucosal fibroids greater than 6 cm in size that become endocavitary may cause postprocedural complications requiring further intervention and medical treatment. PMID- 18430836 TI - Are fibroids that become endocavitary after uterine artery embolization necessarily a complication? AB - OBJECTIVE: This commentary deals with the study by Verma et al. discussing submucosal and endocavitary fibroids after uterine artery embolization (UAE). CONCLUSION: UAE can infarct fibroids. Fibroids spontaneously infarct after childbirth. Because the postpartum cervix is patulous, infarcted fibroids that fall into the uterine cavity easily exit the uterus. Each patient contemplating UAE should anticipate that infarcted fibroids bordering on or inside the uterine cavity may require cervical dilatation or hysteroscopic resection for removal. The addition of either of these two gynecology procedures should not necessarily be regarded as a UAE complication or treatment failure. PMID- 18430837 TI - Pediatric cardiovascular CT angiography: radiation dose reduction using automatic anatomic tube current modulation. AB - OBJECTIVE: The purpose of this study was to assess the effect of weight-based scanning protocols and automatic tube current modulation on the tube current-time product and image quality at pediatric cardiovascular 64-MDCT angiography. MATERIALS AND METHODS: Our pediatric cardiovascular 64-MDCT protocols use a weight-based algorithm to determine nominal tube voltage settings with 80, 100, and 120 kV. Automatic tube current modulation was used for each case. The mAs, volume CT dose index (CTDI(vol)), and dose-length product (DLP) values were recorded and the effective dose calculated. On the basis of the selected nominal tube current, the dose values that would have been delivered without tube current modulation were also calculated. Scans were compared with 16-MDCT using 120 kVp and 120 mAs. Two radiologists independently rated image quality on a 5-point scale. Image noise was objectively measured within four different regions of interest. Findings at CT were clinically correlated with results of cardiac sonography, angiography, or surgery. RESULTS: Thirty-eight 64-MDCT and 30 16-MDCT scans were evaluated. Mean diagnostic quality for 64-MDCT was rated at 3.6 +/- 0.4 and mean image noise was 8.9 +/- 4.5 H. Results with 16-MDCT were not significantly different: diagnostic quality (3.6 +/- 0.4; p = 0.97) and image noise (9.1 +/- 2.8 H; p = 0.31). Scanning with automatic tube current modulation significantly (p < 0.05) reduced the tube current time-product compared with scanning without automatic tube current modulation (-57.8%/54.1/128 mAs) or with 16-MDCT (-47.9%/54.1/104.37 mAs), respectively. The mAs values were significantly (p < 0.05) lower for 80 kVp than for 100 or 120 kVp scans, but image quality and image noise were not significantly (p = 0.24) different. Agreement between MDCT and clinical findings was excellent. CONCLUSION: Under simulated conditions, automatic tube current modulation combined with low tube voltage settings significantly reduced radiation exposure and thus appears preferable in pediatric cardiovascular 64-MDCT. PMID- 18430838 TI - CT of pediatric vascular stents used to treat congenital heart disease. AB - OBJECTIVE: The purpose of our study was to assess the visibility of lumen narrowing of pediatric vascular stents using various CT dose parameters in an in vitro model. MATERIALS AND METHODS: Ten steel stents of varying designs and sizes commonly used in the treatment of congenital heart disease were implanted in polyvinyl chloride (PVC) tubes and three of the 10 stents were partially obstructed with wax by filling 25% (mild) to 60% (moderate) of the lumen with contrast material. On a 64-MDCT scanner, the stents were scanned at tube voltages (kVp) of 80, 100, and 120 and at tube currents (mA) of 40, 80, 120, and 160. CT measurements of inner-stent diameter, strut thickness, and percent lumen (in stent) stenoses were compared with biplane fluoroscopy of digital angiography. RESULTS: The stent diameter and percent stenosis on all CT images were consistently smaller than measured on digital angiography but were highly correlated (r = 0.97; p < 0.0001) with improvement as stent diameter increased (93% agreement with digital angiography for 4-mm stent, up to 99% for 25-mm stent; p = 0.001). Moderate stenosis could be assessed better than mild stenosis (99% vs 91% agreement with digital angiography; p = 0.003). Increasing exposure settings improved CT correlation of all measurements for mA up to 120 and kVp up to 100 (98.1% agreement). Higher settings did not improve accuracy (93.9% for 160 mA at 120 kVp; p = 0.03). CONCLUSION: CT is feasible to assess lumen narrowing of pediatric vascular stents at a wide range of tube settings. The study suggests that it is possible to lower the radiation exposure settings without loss in image quality or accuracy in detecting in-stent stenoses. PMID- 18430839 TI - Prevalence, pattern, and spectrum of glenoid bone loss in anterior shoulder dislocation: CT analysis of 218 patients. AB - OBJECTIVE: The purpose of our study was to determine the prevalence, pattern, and spectrum of glenoid bone loss in anterior shoulder dislocation, to relate this to the frequency of dislocation, and to test the appropriateness of the measurement method. SUBJECTS AND METHODS: Two hundred eighteen patients with single or recurrent anterior shoulder dislocation underwent shoulder CT examination. Fifteen patients had bilateral dislocation. Prevalence and severity of glenoid bone loss and glenoid fracture were assessed. CT examinations of 56 control subjects without shoulder dislocation were evaluated for glenoid contour and side to-side variation in glenoid width. RESULTS: Glenoid bone loss was present in 27 (41%) of 66 patients with first-time unilateral dislocation and 118 (86%) of 137 patients with recurrent unilateral dislocation. Glenoid bone loss ranged from 0.3% to -33% (mean, -10.8% +/- 7.9%). Seventy-four (51%) of 145 patients had < or = 10% glenoid bone loss, 54 (37%) had between 10% and 20%, eight (6%) had between 20% and 25% glenoid bone loss, and nine (6%) had > or = 25% glenoid bone loss. Glenoid rim fractures were present in 49 (21%) of 233 dislocated shoulders. The number of dislocations correlated moderately with the severity of glenoid bone loss (r = 0.56). The normal side-to-side glenoid width variation was small (0.46 +/- 0.81 mm). CONCLUSION: Glenoid bone loss is common in anterior shoulder dislocation. It is probably multifactorial in origin, is usually mild in degree, and has a maximum observed severity of -33%. Dislocation frequency cannot accurately predict the degree of bone loss. PMID- 18430840 TI - Detection of bone graft failure in lumbar spondylodesis: spatial resolution with high-resolution peripheral quantitative CT. AB - OBJECTIVE: In spinal surgery, anterior spondylodesis is often combined with bone grafting, and graft integration is assessed with CT. High-resolution peripheral quantitative CT offers a resolution of 82 mum. The aim of this study was to compare the outcome of anterior spondylodesis as assessed with three radiologic procedures. MATERIALS AND METHODS: Monosegmental lumbar spondylodesis with autologous iliac crest graft or solvent-preserved bovine cancellous bone was performed on seven sheep. The fused spinal segments were explanted after 24 weeks and examined with clinical 64-MDCT, high-resolution peripheral quantitative CT, and contact radiography. In 2D views, the area of the disk space bridged by bone was assessed, and the grafts were examined for fractures. RESULTS: In three of seven sheep, clinical CT erroneously showed stable consolidation, whereas contact radiography revealed a clearly visible graft fracture, as did high-resolution peripheral quantitative CT. There was a statistically significant difference (p = 0.038) between bone volume assessed with clinical CT and that assessed with contact radiography. There was an almost significant difference (p = 0.053) between volumes assessed with high-resolution peripheral quantitative CT and clinical MDCT. CONCLUSION: High-resolution peripheral quantitative CT, a technique approved for clinical use, has higher resolution in imaging of bone structure than does 64-MDCT. Our results show that high-resolution peripheral quantitative CT is superior to 64-MDCT in assessing osseous implant integration after anterior spondylodesis. The specimen size limit, however, prohibits in vivo use of this method in evaluation of the human spine. Our results suggest that in clinical practice, persisting symptoms despite radiologic findings of consolidated spondylodesis may be related to graft failure, which cannot be detected with clinically available methods. PMID- 18430841 TI - Femoroacetabular impingement: can the alpha angle be estimated? AB - OBJECTIVE: Femoroacetabular impingement is an important entity with well described radiographic findings. One of the criteria of the cam type of femoroacetabular impingement is femoral head-neck dysplasia, denoted mathematically as the "alpha angle." Several observers have reported that direct measurement of the angle may not be necessary because subjective appraisal may yield similar results. We sought to scientifically determine the accuracy of a subjective assessment, using the calculated angle as the gold standard. MATERIALS AND METHODS: At 1.5 T, 50 consecutive patients' hips were evaluated on sets of oblique axial images. Two musculoskeletal radiologists recorded their subjective opinion as to the alpha angle using a confidence scale of 1-5. Direct mathematic measurement of the alpha angle was done by a third independent observer and correlated with the subjective results. Correlations between the subjective and measured angles and interobserver variation were calculated. RESULTS: Statistically, significant variability was seen in the subjective assessment of the alpha angle. When the alpha angle was > 55 degrees, the area under the receiver operating characteristic curve (AUC) was 0.606, indicating that visual assessment is a poor predictor of a wide alpha angle. Even in patients with a measured normal alpha angle (< 55 degrees), slightly fewer than half were subjectively thought to possibly, likely, or definitely have abnormal angles. Similarly, more than half of the abnormal cases (alpha angles > 55 degrees) were subjectively thought to possibly or probably be normal. CONCLUSION: Subjective assessment of alpha angles is suboptimal unless one is quite confident of a bone abnormality. PMID- 18430842 TI - Practical experience with sonographically guided phenol instillation of stump neuroma: predictors of effects, success, and outcome. AB - OBJECTIVE: Phantom limb pain and stump pain frequently occur after limb amputation, and stump neuromas play an important role in generation of the pain. The purpose of this study was to evaluate the effects of a previously described optimized procedure for sclerosis of painful stump neuromas under real-time high resolution sonographic guidance. SUBJECTS AND METHODS: In this prospective study, neurosclerosis was performed on 82 patients by means of high-resolution sonographically guided injection of up to 0.8 mL of 80% phenol solution according to a standardized protocol. RESULTS: During treatment all patients had marked improvement in terms of reduction of pain measured with a visual analog scale. Twelve (15%) of the subjects were pain free after one to three treatments, nine of the 12 achieving relief with the initial instillation. At 6-month follow-up evaluation, 52 patients assessed their present pain quantity with a simplified three-step score. Twenty (38%) of the 52 patients reported almost unnoticeable pain, and 33 (64%) reported pain equal to the minimum reached during therapy. In 18 (35%) of the 52 patients, the incidence of painful periods had markedly decreased. The neurosclerosis procedure had a low complication rate (5% rate of minor complications, 1.3% rate of major complications). CONCLUSION: The high resolution sonographically guided neurosclerosis procedure had a significantly better outcome than other documented treatments. Sonographically guided neurosclerosis should be included in the management of chronic phantom limb and stump pain. PMID- 18430843 TI - 3-T MRI of rectal carcinoma: preoperative diagnosis, staging, and planning of sphincter-sparing surgery. AB - OBJECTIVE: The purpose of this study was to assess the accuracy of 3-T MRI in the preoperative diagnosis, staging, and planning of surgical management of rectal carcinoma. SUBJECTS AND METHODS: Thirty-eight patients (23 men, 15 women) with clinically suspected rectal carcinoma underwent 3-T MRI. Coronal, axial, and sagittal T2-weighted sequences with and without fat suppression; axial T1 weighted spin-echo sequences; axial T1-weighted gradient-echo sequences with and without fat suppression; oblique 2D MR hydrography; and 3D fat-suppressed dynamic contrast-enhanced MRI were performed. Image quality with these sequences was evaluated by three radiologists experienced in body MRI. The significance of difference in results with the sequences was tested. The manner in which MRI staging and feasibility of sphincter-sparing surgery agreed with operative and pathologic findings was evaluated with kappa statistics. RESULTS: Rectal carcinoma was identified on MRI and confirmed histologically in all 38 patients. MRI findings were correctly predictive of T category in 35 cases (accuracy, 92.1%). In 31 (96.9%) of 32 resectable cases,sphincter-sparing surgical approaches were accurately chosen on the basis of MRI findings. Among the 11 sequences, 3D fat-suppressed dynamic contrast-enhanced MRI best delineated tumor margins. Coronal and axial T2-weighted images also well depicted tumor margins with minimal artifact. T2-weighted images were superior to unenhanced T1-weighted images. CONCLUSION: MRI of rectal cancer at 3 T is accurate for prediction of T category and the feasibility of sphincter-sparing surgery. The best images were obtained with coronal, sagittal, and axial T2-weighted sequences and 3D fat suppressed dynamic contrast-enhanced MRI. PMID- 18430844 TI - Protrusion method for automated estimation of polyp size on CT colonography. AB - OBJECTIVE: The purpose of this study was to assess the accuracy and measurement variability of automated lesion measurement on CT colonography in comparison with manual 2D and 3D techniques under varying scanning conditions. MATERIALS AND METHODS: The study included phantoms (23 phantom objects) and patients (16 polyps). Measurement with sliding calipers served as the reference for the phantom data. The mean of two independent colonoscopic measurements was the reference for the polyps. The automated measurement was developed for a computer aided detection scheme, and the size of any detected object was obtained from measurement of its largest diameter. The automated measurement was compared with manual 2D and 3D measurements by two experienced observers. RESULTS: For phantom data, the measurement variability of the automated method was significantly less than that of the two observers (p < 0.05), except for the 3D measurement by observer 1, as follows: automated, 0.86 mm; observer 1, 1.76 mm (2D), 0.96 (3D); observer 2, 1.34 mm (2D), 1.45 mm (3D). The variability of the automated method did not differ significantly from that of manual methods in measurement with patient data. The automated method had a systematic error for phantom data (1.9 mm). CONCLUSION: For phantoms, the automated method has less measurement variability than manual 2D and 3D techniques. For true polyps, the measurement variability of the automated method is comparable with that of manual methods. The automated method does not suffer from intraobserver variability. Because systematic error can be calibrated, automated size measurement may contribute to a practical evaluation strategy. PMID- 18430845 TI - Jejunal diverticulosis: findings on CT in 28 patients. AB - OBJECTIVE: The purpose of our study was to better characterize the CT findings of jejunal diverticulosis by retrospectively reviewing abdominal CT scans of 28 patients with this condition on barium examinations. CONCLUSION: Jejunal diverticula have characteristic findings on CT, appearing as discrete round or ovoid, contrast-, fluid-, or air-containing structures outside the expected lumen of the small bowel, with a smooth, barely discernible wall and no recognizable small-bowel folds. Not infrequently, these structures are seen to communicate directly with an adjoining small-bowel loop, a feature best recognized by scrolling the images. Our experience suggests that jejunal diverticulosis can often be recognized on the basis of the characteristic CT features of this condition. PMID- 18430846 TI - Vascular malformation and hemangiomatosis syndromes: spectrum of imaging manifestations. AB - OBJECTIVE: The purpose of this review is to describe the role of imaging and associated findings in the diagnosis of blue rubber bleb nevus syndrome, Proteus syndrome, Klippel-Trenaunay syndrome, and Kasabach-Merritt syndrome. CONCLUSION: Blue rubber bleb nevus, Proteus, Klippel-Trenaunay, and Kasabach-Merritt syndromes are a diverse group of vascular malformation and hemangiomatosis syndromes. Both cutaneous and visceral vascular lesions are associated with these disorders. Accurate diagnosis of these syndromes is important because they can be associated with serious complications, including life-threatening hemorrhage. PMID- 18430847 TI - Diagnosing acute appendicitis in adults: accuracy of color Doppler sonography and MDCT compared with surgery and clinical follow-up. AB - OBJECTIVE: The objective of our study was to evaluate the accuracy of color Doppler sonography and contrast-enhanced MDCT in the diagnosis of acute appendicitis in adults and their utility as a triage tool in lower abdominal pain. MATERIALS AND METHODS: We reviewed the medical records of 420 consecutive adult patients, 271 women and 149 men, 18 years old or older, referred from the emergency department to sonography examination for clinically suspected acute appendicitis between January 2003 and June 2006. Patients underwent sonography of the right upper abdomen and pelvis followed by graded compression and color Doppler sonography of the right lower quadrant. CT was performed in 132 patients due to inconclusive sonography findings or a discrepancy between the clinical diagnosis and the sonography diagnosis. Sonography and CT reports were compared with surgery or clinical follow-up as the reference standard. Statistical analyses were performed by Pearson's chi-square test and cross-tabulation software. RESULTS: Sonography and CT correctly diagnosed acute appendicitis in 66 of 75 patients and in 38 of 39 patients, respectively, and correctly denied acute appendicitis in 312 of 326 and in 92 of 92 patients. Sonography was inconclusive in 17 of 418 cases and CT, in one of 132 cases. Sonography and CT allowed alternative diagnoses in 82 and 42 patients, respectively. Sensitivity, specificity, positive predictive value, negative predictive value, and accuracy for sonography were 74.2%, 97%, 88%, 93%, and 92%, respectively, and for CT, 100%, 98.9%, 97.4%, 100%, and 99%. CONCLUSION: Sonography should be the first imaging technique in adult patients for the diagnosis of acute appendicitis and triage of acute abdominal pain. CT should be used as a complementary study for selected cases. PMID- 18430848 TI - Cost-effectiveness and patient tolerance of low-attenuation oral contrast material: milk versus VoLumen. AB - OBJECTIVE: The purpose of our study was to prospectively compare the cost, effectiveness, and patient tolerance of milk and VoLumen, a 0.1% barium suspension, in patients undergoing abdominal and pelvic CT with oral and i.v. contrast media. SUBJECTS AND METHODS: Two hundred fifteen consecutive outpatients were randomly assigned to receive either whole milk (n = 115) or VoLumen (n = 100). Results were independently reviewed by two radiologists who were blinded to the oral contrast agent used. Degree of bowel distention was qualitatively scored on a 4-point scale, and bowel wall visibility was graded qualitatively on a yes or-no basis. A questionnaire regarding oral contrast tolerability was provided to each patient. Cost comparison of the two agents was performed. RESULTS: No statistically significant differences were seen between whole milk and VoLumen with respect to degree of bowel distention and mural visualization for all segments of bowel studied (p > 0.05 for both reviewers). Significantly more patients ranked milk as pleasant in taste compared with VoLumen (p < 0.0001). More patients preferred milk compared with VoLumen (p < 0.0001). Milk was better tolerated than VoLumen, with fewer abdominal side effects, including abdominal discomfort (p = 0.019), cramping (p = 0.019), nausea (p = 0.016), and diarrhea (p = 0.0002). The cost per patient for VoLumen is $18 compared with $1.48 for milk. CONCLUSION: Whole milk is comparable to VoLumen with respect to bowel distention and bowel wall visualization and has a lower cost, better patient acceptance, and fewer adverse symptoms. Milk is a cost-effective alternative to VoLumen as a low attenuation oral contrast agent. PMID- 18430849 TI - CT laparoscopy for detecting small superficial metastatic lesions of the liver surface: initial experience. AB - OBJECTIVE: The purpose of this study was to determine the efficacy of CT laparoscopy in the detection of superficial metastasis of the liver surface. SUBJECTS AND METHODS: From April 1, 2007, to July 1, 2007, a total of 34 consecutively registered patients (19 men, 15 women; median age, 55 years) with various intraabdominal malignant tumors underwent preoperative CT and composed the study population. All patients underwent superparamagnetic iron oxide enhanced MRI and portal phase contrast-enhanced 64-MDCT, including CT laparoscopy. CT laparoscopy is a form of volume-rendering 3D imaging of the liver that depicts the liver surface in detail. RESULTS: Among 23 patients who underwent surgery for management of a primary tumor, four patients had seven superficial metastatic lesions of the liver surface. None of these lesions had been detected with preoperative axial CT or superparamagnetic iron oxide-enhanced MRI. In contrast, CT laparoscopy revealed four of seven lesions in four patients. On a lesion-by-lesion basis, the sensitivity was 57%, the positive predictive value was 100%, and the accuracy was 57%. CONCLUSION: Our initial experience proves that CT laparoscopy is a promising method for detecting small superficial metastatic lesions of the liver surface. The findings can influence decisions regarding tumor resectability. PMID- 18430850 TI - Effect of T1 shortening on T2-weighted MRI sequences: comparison of hepatic mass conspicuity on images acquired before and after gadolinium enhancement. AB - OBJECTIVE: The purpose of this study was to compare the conspicuity of hepatic lesions on T2-weighted fast-recovery fast spin-echo MR images obtained before and after administration of gadolinium. MATERIALS AND METHODS: We reviewed T2 weighted fast-recovery fast spin-echo images before and after gadolinium enhancement for 84 patients with 118 focal liver lesions. Solid lesions (22 hepatomas, seven ablated hepatomas, 12 metastatic lesions, six cases of focal nodular hyperplasia, five dysplastic nodules, one adenoma) were proved pathologically or with multiple follow-up studies. Nonsolid lesions were diagnosed as hemangiomas (n = 33) or cysts (n = 32) on the basis of imaging features. Two blinded radiologists interpreted the images independently, reading unenhanced images first and gadolinium-enhanced images at least 2 weeks later. Lesion conspicuity was ranked as follows: 1, poor; 2, moderate; 3, good; 4, excellent. The sign test was used for qualitative scoring of imaging pairs (unenhanced and gadolinium enhanced). The Fisher's exact test was used for subgroup analysis of solid and nonsolid lesions. RESULTS: On gadolinium-enhanced T2-weighted images, 21 (17.8%) of 118 of the lesions had improved conspicuity, 86 (72.9%) had no difference in conspicuity, and 11 (9.3%) appeared worse. No statistically significant difference was found between unenhanced and enhanced images (p = 0.11), but a trend toward improved conspicuity with gadolinium enhancement was observed. Subgroup analysis showed that on gadolinium-enhanced T2 weighted images, visualization of solid hepatic lesions (28.3%) was significantly better than that of nonsolid lesions (9.2%) (p = 0.01). CONCLUSION: Compared with unenhanced T2-weighted images, gadolinium-enhanced T2-weighted images had a trend toward improved conspicuity of focal liver lesions. Subgroup analysis showed that visualization of solid lesions benefited significantly more from use of gadolinium-enhanced T2-weighted sequences than did visualization of nonsolid lesions. PMID- 18430851 TI - Planning sonography to assess the feasibility of percutaneous radiofrequency ablation of hepatocellular carcinomas. AB - OBJECTIVE: The objective of our study was to evaluate the feasibility rate and the reasons for infeasibility of percutaneous radiofrequency ablation of hepatocellular carcinomas (HCCs) at planning sonography. MATERIALS AND METHODS: We retrospectively evaluated 248 patients who had been referred for planning sonography at our department between October 2005 and February 2006. We reviewed the radiologic reports of 256 planning sonography examinations in terms of the rate of feasible cases and the reasons for infeasibility. The reasons for infeasibility were classified in six categories: an inconspicuous tumor, an inadequate electrode path, an organ vulnerable to collateral thermal damage, a tumor that was too large or too many tumors, and a high risk of the heat sink effect. In addition, we assessed the treatment modality for the patients who were determined to have HCCs for which percutaneous radiofrequency ablation was not feasible. RESULTS: In 141 (55%) of the 256 planning sonography examinations, percutaneous radiofrequency ablation was feasible. The remaining 115 (45%) planning sonography examinations revealed that radiofrequency ablation was not a feasible procedure. The reasons for infeasibility included an inconspicuous tumor in 77 patients (55.8%), an inadequate electrode path in 33 patients (23.9%), an organ vulnerable to collateral thermal damage in 14 patients (10.1%), a tumor that was too large or too many tumors in eight patients (5.8%), a high risk of the heat sink effect in five patients (3.6%), and a portal vein thrombosis in one patient (0.7%). One reason for infeasibility was found in 96 patients, two reasons in 19 patients, and four reasons in one patient. Seventy (61.9%) of 113 patients for whom radiofrequency ablation was not feasible underwent transcatheter arterial chemoembolization as an alternative treatment. CONCLUSION: In approximately half of the patients for whom percutaneous radiofrequency ablation of HCC is requested, the procedure is not feasible, mainly due to inconspicuous tumors, at planning sonography. Additional objective criteria for assessing the feasibility of radiofrequency ablation and therapeutic strategies according to the reasons for infeasibility should be investigated further. PMID- 18430852 TI - Evaluation of the severity of chronic hepatitis C with 3-T1H-MR spectroscopy. AB - OBJECTIVE: The purpose of this study was to compare the spectral characteristics of lipids, choline-containing compounds, and glutamine-glutamate complex assessed with (1)H-MR spectroscopy with the histologic findings in patients with chronic hepatitis C. SUBJECTS AND METHODS: Nine healthy controls and 30 patients with biopsy-proven hepatitis C virus-related liver disease participated in this prospective study. Degree of fibrosis and histologic activity were scored according to the METAVIR classification. The percentage of involved hepatocytes was used to grade steatosis. Hepatic spectra were obtained with a 3-T spectroscopic system. Tenfold cross-validated stepwise discriminant analysis was performed to classify disease severity on the basis of the spectroscopic findings. RESULTS: There was a strong correlation between (1)H-MR spectroscopically measured lipid concentration and the degree of steatosis at histologic examination (r = 0.9236, p < 0.0001). This finding enabled clear separation of groups according to degree of histologically determined steatosis. Variation in lipid concentration was consistent with the degree of steatosis (r = 0.7265, p < 0.0001) and stage of fibrosis (r = 0.8156, p < 0.0001). In univariate analysis, concentrations of both choline-containing compounds and glutamine glutamate complex had a direct correlation with histologic grade (p < 0.0001) and degree of steatosis (p < 0.0001) but not with stage of fibrosis (p > 0.05). In multivariate analysis, the only factor independently associated with concentrations of choline-containing compounds and glutamine-glutamate complex was histologic grade. In cross-validated discriminant analysis based on choline containing compound, glutamine-glutamate complex, and lipid resonance, 70% (21 of 30) of the histologic grade groups and 73% (22 of 30) of the steatosis groups were correctly classified. CONCLUSION: Hydrogen-1 MR spectroscopy can be an alternative to liver biopsy in the evaluation of steatosis and necroinflammatory activity in liver disease but is not useful for complete evaluation of hepatic fibrosis. PMID- 18430853 TI - Transcatheter arterial chemoembolization in patients with hepatocellular carcinoma on the waiting list for orthotopic liver transplantation. AB - OBJECTIVE: The objective of this study was to perform a retrospective analysis of patients with hepatocellular carcinoma (HCC) who underwent transcatheter arterial chemoembolization (TACE) before undergoing liver transplantation at our institution. SUBJECTS AND METHODS: From January 2000 to August 2005, 56 patients with HCC underwent TACE before orthotopic liver transplantation (OLT). Radiologic findings before and after TACE were assessed and correlated with histologic findings after OLT. The area of induced necrosis was pathologically evaluated in each HCC nodule. RESULTS: One hundred thirty-one HCC nodules were detected at histologic study. One hundred seventeen HCC nodules (91.4%) were hyperenhancing in the arterial phase on the preoperative imaging studies. The percentage of tumor necrosis was greater than 90% in 48 nodules (38%), between 50% and 90% in 19 nodules (15%), and less than 50% in 61 nodules (48%); tumor necrosis data were not recorded for the remaining three nodules. The size of the preoperatively detected lesions ranged from 0.2 to 9 cm (mean, 2.58 cm). The mean percentage of tumor necrosis was 67.8% in this group, but it rose to 79.2% in the hypervascular lesions. The size of the nodules that were not detected preoperatively ranged from 0.1 to 1.9 cm (mean, 0.68 cm), and the mean percentage of tumor necrosis was only 1.57%. CONCLUSION: TACE is a safe treatment in well-selected patients. Its antitumoral effect is high in hypervascular lesions (mean necrosis, 79.2%). It provides good local control in preoperatively diagnosed HCC (mean necrosis, 67.8%), but its impact is limited in lesions not detected preoperatively (mean necrosis, 1.57%). PMID- 18430854 TI - Multicenter randomized controlled trial of the costs and effects of noninvasive diagnostic imaging in patients with peripheral arterial disease: the DIPAD trial. AB - OBJECTIVE: The purpose of our study was to compare the costs and effects of three noninvasive imaging tests as the initial imaging test in the diagnostic workup of patients with peripheral arterial disease. MATERIALS AND METHODS: Of 984 patients assessed for eligibility, 514 patients with peripheral arterial disease were randomized to MR angiography (MRA) or duplex sonography in three hospitals and to MRA or CT angiography (CTA) in one hospital. The outcome measures included the clinical utility, functional patient outcomes, quality of life, and actual diagnostic and therapeutic costs related to the initial imaging test during 6 months of follow-up. RESULTS: With adjustment for potentially predictive baseline variables, the learning curve, and hospital setting, a significantly higher confidence and less additional imaging were found for MRA and CTA compared with duplex sonography. No statistically significant differences were found in improvement in functional patient outcomes and quality of life among the groups. The total costs were significantly higher for MRA and duplex sonography than for CTA. CONCLUSION: The results suggest that both CTA and MRA are clinically more useful than duplex sonography and that CTA leads to cost savings compared with both MRA and duplex sonography in the initial imaging evaluation of peripheral arterial disease. PMID- 18430855 TI - Assessment of global right ventricular function on 64-MDCT compared with MRI. AB - OBJECTIVE: The aim of this study was to compare ECG-gated 64-MDCT with MRI for the assessment of global right ventricular (RV) function from coronary CT angiography data. SUBJECTS AND METHODS: Thirty-eight patients (25 men, 13 women; mean age +/- SD, 55.0 +/- 8.8 years) with suspected coronary artery disease underwent contrast-enhanced 64-MDCT (64 x 0.6 mm, 120 kV, 770 mAs(eff)) and 1.5-T MRI (balanced fast-field echo; TR/TE, 3.3/1.6; flip angle, 60 degrees ; 50 phases). Double oblique short-axis MDCT and MR images were used for further analysis. End-diastolic volume (EDV), end-systolic volume (ESV), stroke volume (SV), and ejection fraction (EF) were computed from manually drawn endocardial contours of the right ventricle. For statistical analysis, repeated-measures analysis of variance and Pearson's correlation coefficients were calculated. Bland-Altman plots were computed. RESULTS: In general, RV volumes calculated from 64-MDCT agreed well with those calculated from MRI. The mean EF (+/- SD) calculated from MDCT and MRI was 51.0% +/- 7.8% and 51.4% +/- 7.3%, respectively. An excellent correlation was observed for EDV (r = 0.99), ESV (r = 0.98), SV (r = 0.98), and EF (r = 0.97). Bland-Altman plots showed no systematic variation between MDCT and MRI data. No statistically significant differences (p < or = 0.05) between the techniques were found. CONCLUSION: Although contrast injection is optimized for visualization of the coronary arteries, retrospectively ECG gated 64-MDCT permits reliable assessment of global RV function. PMID- 18430856 TI - Ground-glass opacities on thin-section helical CT: differentiation between bronchioloalveolar carcinoma and atypical adenomatous hyperplasia. AB - OBJECTIVE: The purpose of our study was to investigate the differentiation between bronchioloalveolar carcinoma and atypical adenomatous hyperplasia manifesting pure ground-glass opacity (GGO) based on selected features on thin section helical CT scans. MATERIALS AND METHODS: We evaluated 35 bronchioloalveolar carcinomas and 17 atypical adenomatous hyperplasias that were histologically confirmed and that manifested pure GGO on thin-section helical CT scans. We recorded the age, sex, and smoking history (Brinkman index) of the patients. Two board-certified radiologists measured the maximum diameter and mean attenuation value of the nodules; the measured values were averaged for each nodule. Using a 3-point scale, they visually assessed the images for consensus with respect to nodular sphericity, marginal irregularity, vascular convergence, pleural retraction, and findings of an internal air bronchogram. CT findings of atypical adenomatous hyperplasia and bronchioloalveolar carcinoma were compared using univariate and multivariate logistic regression analysis; the odds ratio was computed using the atypical adenomatous hyperplasia group as the reference group. RESULTS: By univariate analysis, the patient age, nodular maximum diameter, mean attenuation value, and findings of an internal air bronchogram were statistically significantly associated with bronchioloalveolar carcinoma (odds ratio [OR] = 1.10 [p = 0.012], OR = 1.27 [p < 0.01], OR = 1.01 [p = 0.023], and OR = 25.30 [p < 0.001], respectively), and sphericity was significantly associated with atypical adenomatous hyperplasia (OR = 0.059, p < 0.001). By multivariate analysis, sphericity was significantly associated with atypical adenomatous hyperplasia (OR = 0.125, p = 0.042) and findings of an internal air bronchogram were associated with bronchioloalveolar carcinoma (OR = 16.10, p = 0.007). CONCLUSION: Nodular sphericity and an internal air bronchogram were useful at thin-section helical CT performed to differentiate between bronchioloalveolar carcinoma and atypical adenomatous hyperplasia. Interobserver agreement was high for each finding. PMID- 18430857 TI - Potential role of diffusion tensor MRI in the differential diagnosis of mild cognitive impairment and Alzheimer's disease. AB - OBJECTIVE: The purpose of this study was to evaluate the fractional anisotropy values of several white matter tracts with the aim of differentiating a healthy population from persons with mild cognitive impairment or Alzheimer's disease. SUBJECTS AND METHODS: Seventy-nine patients with memory impairment and 16 volunteer controls participated in the study. MRI was performed with a 1.5-T system. Conventional MR images and diffusion tensor images were obtained for all participants. The diffusion tensor imaging data were postprocessed, and low b value, fractional anisotropy, and fractional anisotropy color-coded maps were calculated. With the three maps as an anatomic reference, fractional anisotropy was measured for hippocampal formations, superior longitudinal fascicles, posterior cingulate gyri, and the splenium of the corpus callosum. Kruskal-Wallis and Steel-type multiple-comparison nonparametric tests were performed for the statistical analysis. RESULTS: The fractional anisotropy values for the splenium of the corpus callosum, bilateral posterior cingulate gyri, and bilateral superior longitudinal fascicles of patients with mild cognitive impairment and those with probable Alzheimer's disease were significantly lower than the values of controls. No differences were found in hippocampal formations in any group. No significant difference was found in fractional anisotropy values in comparisons of mild cognitive impairment versus possible Alzheimer's disease and probable Alzheimer's disease or comparisons of probable Alzheimer's disease and possible Alzheimer's disease. CONCLUSION: Diffusion tensor imaging is a promising technique for the evaluation of patients with probable mild cognitive impairment. Early detection of the disease expands the treatment options, increasing the likelihood of a good clinical response and enhancing the quality of life of patients and their relatives. Further studies with larger populations are needed to confirm the role of diffusion tensor imaging in the evaluation of memory impairment. PMID- 18430858 TI - Usefulness of laryngeal phonation CT in the diagnosis of vocal cord paralysis. AB - OBJECTIVE: The purpose of this study was to determine the effectiveness of laryngeal phonation CT for the diagnosis of vocal cord paralysis by examining the physiologic and functional changes in the larynx during vowel phonation in patients with vocal cord paralysis. SUBJECTS AND METHODS: For the control study, three healthy volunteers underwent laryngeal phonation CT while vocalizing the vowels /hee/, /ih/, and /ah/, and reconstructed coronal images of the larynx were obtained. After the control study, 28 patients with unilateral vocal cord paralysis underwent laryngeal phonation CT during /hee/ phonation, which was chosen as a most appropriate vowel for this purpose. Changes in the paralyzed and normal vocal cords were evaluated quantitatively and qualitatively on coronal reconstruction images. RESULTS: On the coronal reconstructed images from the healthy volunteers, the normal cords had a shoulder formation appearance, and the cords lay within 1 mm lateral to the midline during phonation. For patients with vocal cord paralysis during /hee/ phonation, the average angle formed by the long axis of the vocal cord and the midline was 71.67 degrees on the affected side and 92.21 degrees on the normal side (p = 0.001). The vocal cord edges lay 1.5 mm lateral to the midline on the affected side and 0.44 mm lateral to the midline on the normal side (p = 0.003). In the qualitative study, the two observers found the coronal reconstructions of the laryngeal phonation CT scans yielded a higher detection rate than did conventional axial CT. CONCLUSION: Laryngeal phonation CT proved more useful for evaluating vocal cord paralysis than did conventional CT and can be used as a primary diagnostic tool when vocal cord paralysis is suspected. PMID- 18430859 TI - Postmortem whole-body CT angiography: evaluation of two contrast media solutions. AB - OBJECTIVE: The objective of our study was to establish a standardized procedure for postmortem whole-body CT-based angiography with lipophilic and hydrophilic contrast media solutions and to compare the results of these two methods. MATERIALS AND METHODS: Minimally invasive postmortem CT angiography was performed on 10 human cadavers via access to the femoral blood vessels. Separate perfusion of the arterial and venous systems was established with a modified heart-lung machine using a mixture of an oily contrast medium and paraffin (five cases) and a mixture of a water-soluble contrast medium with polyethylene glycol (PEG) 200 in the other five cases. Imaging was executed with an MDCT scanner. RESULTS: The minimally invasive femoral approach to the vascular system provided a good depiction of lesions of the complete vascular system down to the level of the small supplying vessels. Because of the enhancement of well-vascularized tissues, angiography with the PEG-mixed contrast medium allowed the detection of tissue lesions and the depiction of vascular abnormalities such as pulmonary embolisms or ruptures of the vessel wall. CONCLUSION: The angiographic method with a water soluble contrast medium and PEG as a contrast-agent dissolver showed a clearly superior quality due to the lack of extravasation through the gastrointestinal vascular bed and the enhancement of soft tissues (cerebral cortex, myocardium, and parenchymal abdominal organs). The diagnostic possibilities of these findings in cases of antemortem ischemia of these tissues are not yet fully understood. PMID- 18430860 TI - Detection of smuggled cocaine in cargo using MDCT. AB - OBJECTIVE: Smuggling dissolved drugs, especially cocaine, in bottled liquids is an ongoing problem at borders. Common fluoroscopy of packages at the border cannot detect contaminated liquids. The objective of our study was to develop an MDCT screening method to detect cocaine-containing vessels that are hidden between uncontaminated ones in a shipment. MATERIALS AND METHODS: Studies were performed on three wine bottles containing cocaine solutions that were confiscated at the Swiss border. Reference values were obtained by scans of different sorts of commercially available wine and aqueous solutions of dissolved sugar. All bottles were scanned using MDCT, and data evaluation was performed by measuring the mean peak of Hounsfield units. To verify the method, simulated testing was performed. RESULTS: Using measurements of the mean peak of Hounsfield units enables the detection of dissolved cocaine in wine bottles in a noninvasive and rapid fashion. Increasing opacity corresponds well with the concentration of dissolved cocaine. Simulated testing showed that it is possible to distinguish between cocaine-contaminated and uncontaminated wine bottles. CONCLUSION: The described method is an efficacious screening method to detect cocaine contaminated bottles that are hidden between untreated bottles in cargo. The noninvasive examination of cargo allows a questionable delivery to be tracked without arousing the suspicion of the smugglers. PMID- 18430861 TI - An introduction to the Fourier transform: relationship to MRI. AB - OBJECTIVE: The Fourier transform, a fundamental mathematic tool widely used in signal analysis, is ubiquitous in radiology and integral to modern MR image formation. Understanding MRI techniques requires a basic understanding of what the Fourier transform accomplishes. MR image encoding, filling of k-space, and a wide spectrum of artifacts are all rooted in the Fourier transform. CONCLUSION: This article illustrates these basic Fourier principles and their relationship to MRI. PMID- 18430862 TI - Method for decreasing uptake of 18F-FDG by hypermetabolic brown adipose tissue on PET. AB - OBJECTIVE: The purpose of this study was to determine whether use of a high-fat, very-low-carbohydrate protocol for preparing patients for PET decreases the frequency of (18)F-FDG uptake by hypermetabolic brown adipose tissue (BAT) on PET scans. MATERIALS AND METHODS: In this HIPAA-compliant retrospective study, 741 FDG PET/CT scans obtained during the winter months (October 1-April 30) for patients who prepared with a high-fat, very-low-carbohydrate, protein-permitted protocol were compared with 1,229 FDG PET scans obtained during the winter months for patients who prepared by fasting. FDG uptake on PET scans co-localized with regions of fat identified on the CT scans was assumed to represent hypermetabolic BAT. The categoric variables frequency of occurrence of hypermetabolic BAT (present or not) and the sex ratios of the groups before and after the change in preparation were compared by use of a chi-square test. The continuous variables of age and blood glucose level were compared by use of a two-tailed Student's t test. RESULTS: In this intention-to-treat analysis, there was no difference between the fasting (n = 1,229) and the high-fat, very-low-carbohydrate, protein permitted diet (n = 741) groups in terms of age and sex. Patients who prepared with the high-fat diet had a significantly lower frequency of hypermetabolic BAT uptake on FDG PET scans during the winter months (p<0.0002) and had lower blood glucose levels (p<<0.001). CONCLUSION: In this intention-to-treat analysis, use of a high-fat preparation protocol significantly lowered the frequency of uptake of FDG by hypermetabolic BAT on FDG PET studies. Use of this protocol has the potential to decrease the rate of false-positive findings on oncologic FDG PET scans. PMID- 18430863 TI - Transposition of three amino acids transforms the human metabotropic glutamate receptor (mGluR)-3-positive allosteric modulation site to mGluR2, and additional characterization of the mGluR2-positive allosteric modulation site. AB - Glutamate is a major neurotransmitter in the central nervous system, and abnormal glutamate neurotransmission has been implicated in many neurological disorders, including schizophrenia, Alzheimer's disease, Parkinson's disease, addiction, anxiety, depression, epilepsy, and pain. Metabotropic glutamate receptors (mGluRs) activate intracellular signaling cascades in a G protein-dependent manner, which offer the opportunity for developing drugs that regulate glutamate neurotransmission in a functionally selective manner. In the present study, we further characterize the human mGluR2 (hmGluR2) potentiator binding site by showing that the substitution of the three amino acids found to be required for hmGluR2 potentiation, specifically Ser(688), Gly(689), and Asn(735), with the homologous hmGluR3 amino acids, inactivates the positive allosteric modulator activity of several structurally unique mGluR2 potentiators. Based on the characterization of the hmGluR2 potentiator binding site, we developed a novel scintillation proximity assay that was able to discriminate between compounds that were hmGluR2-specific potentiators, and those that were active on both hmGluR2 and hmGluR3. In addition, we substituted Ser(688), Gly(689), and Asn(735) into hmGluR3 and created an active hmGluR2 allosteric modulation site on the hmGluR3 receptor. PMID- 18430865 TI - Kinin B2 receptor-mediated bradykinin internalization and metalloendopeptidase EP24.15-dependent intracellular bradykinin degradation. AB - Kinins are potent proinflammatory peptides that are produced extracellularly and are rapidly degraded by extracellular peptidases and by intracellular peptidases accessed by kinins via receptor-mediated endocytosis. In this study, we developed model cell systems expressing the kinin B(2) receptor (B(2)R) and the metalloendopeptidase thimet oligopeptidase (EC 3.4.24.15; EP24.15) either individually or together to address 1) the cellular and functional relationship between these proteins and 2) the participation of EP24.15 in the metabolism of bradykinin (BK) after BK internalization via B(2)R. B(2)R was localized almost exclusively in the plasma membrane, whereas EP24.15 was localized both intracellularly and on the cell surface and secreted in the media. Intracellular EP24.15 was present throughout the cell, both cytosolic and particulate, with less nuclear localization and no colocalization with either the endoplasmic reticulum marker calnexin or the Golgi marker GM130. No direct colocalization of B(2)R and EP24.15 was observed using immunofluorescence microscopy. However, the two proteins coimmunoprecipitated specifically, and EP24.15 attenuated maximal B(2)R responsiveness without influencing the potency of BK to stimulate phosphoinositide hydrolysis and intracellular Ca(2+) mobilization. Cell surface bound BK remained intact in cells overexpressing EP24.15 but was degraded intracellularly in an EP24.15-dependent manner upon B(2)R-mediated endocytosis. These results show that EP24.15 acts to negatively regulate B(2)R responsiveness, and it serves as an intracellular peptidase in the degradation of BK specifically internalized via this receptor. PMID- 18430864 TI - Effect of cysteine mutagenesis on the function and disulfide bond formation of human ABCG2. AB - ABCG2 is a member of the ATP-binding cassette (ABC) transporter superfamily. Its overexpression causes multidrug resistance in cancer chemotherapy. Based on its apparent half size in sequence when compared with other traditional ABC transporters, ABCG2 has been thought to exist and function as a homodimer linked by intermolecular disulfide bonds. However, recent evidence suggests that ABCG2 may exist as a higher form of oligomers due to noncovalent interactions. In this study, we attempted to create a cysless mutant ABCG2 as a tool for further characterization of this molecule. However, we found that the cysless mutant ABCG2 is well expressed but not functional. Mapping of the cysteine residues showed that three cysteine residues (Cys284, Cys374, and Cys438) are required concurrently for the function of ABCG2 and potentially for intramolecular disulfide bond formation. We also found that the cysteine residues (Cys592, Cys603, and Cys608) in the third extracellular loop are involved in forming intermolecular disulfide bonds and that mutation of these residues does not affect the expression or drug transport activity of human ABCG2. Thus, we conclude that Cys284, Cys374, and Cys438, which may be involved in intramolecular disulfide bond formation, are concurrently required for ABCG2 function, whereas Cys592, Cys603, and Cys608, potentially involved in intermolecular disulfide bond formation, are not required. PMID- 18430866 TI - Evaluation of the association of IGF2BP2 variants with type 2 diabetes in French Caucasians. AB - OBJECTIVE: We performed a comprehensive genetic association study of common variation spanning the IGF2BP2 locus in order to replicate the association of the "confirmed" type 2 diabetes susceptibility variants rs4402960 and rs1470579 in the French Caucasian population and to further characterize the susceptibility variants at this novel locus. RESEARCH DESIGN AND METHODS: We genotyped a total of 21 tagging single nucleotide polymorphisms spanning the IGF2BP2 locus in our type 2 diabetes case-control cohort comprising 3,093 French Caucasian subjects. RESULTS: IGF2BP2 variants rs4402960 and rs1470579 were not associated with type 2 diabetes in the present study (P = 0.632 and P = 0.896, respectively). Meta analysis of genotype data from over 34,000 subjects demonstrated that our inability to replicate rs4402960/rs1470579 was consistent with the findings from several previous genome-wide association study (GWAS) datasets that were underpowered to detect this modest association signal (odds ratio [OR] 1.14). We obtained novel evidence that rs9826022, a borderline rare variant (5% minor allele frequency) in the 3' downstream region, was associated with type 2 diabetes (P = 0.0002; OR 1.53 [95% CI 1.22-1.91]). This result was corroborated by the meta-analysis of 10,542 genotypes from the current study and GWAS datasets using both fixed (P = 9.47 x 10(-6); 1.30 [1.16-1.46]) and random effects (P = 0.001; 1.30 [1.11-1.52)] calculations. CONCLUSIONS: We were unable to replicate the confirmed rs4402960/rs1470579 susceptibility variants but found novel evidence for a rare variant in the 3' downstream region of IGF2BP2. Further genetic and functional studies are required to identify the etiological IGF2BP2 variants. PMID- 18430868 TI - Can MR imaging be used to predict the degree and prognosis of peripheral nerve injuries? PMID- 18430870 TI - Net Assets: personal technology for productivity in radiology. PMID- 18430869 TI - The self-portraits of Frida Kahlo. PMID- 18430871 TI - MR Imaging of hepatocellular carcinoma in the cirrhotic liver: challenges and controversies. AB - The incidence of hepatocellular carcinoma (HCC) is expected to increase in the next 2 decades, largely due to hepatitis C infection and secondary cirrhosis. HCC is being detected at an earlier stage owing to the implementation of screening programs. Biopsy is no longer required prior to treatment, and diagnosis of HCC is heavily dependent on imaging characteristics. The most recent recommendations by the American Association for the Study of Liver Diseases (AASLD) state that a diagnosis of HCC can be made if a mass larger than 2 cm shows typical features of HCC (hypervascularity in the arterial phase and washout in the venous phase) at contrast material-enhanced computed tomography or magnetic resonance (MR) imaging or if a mass measuring 1-2 cm shows these features at both modalities. There is an ever-increasing demand on radiologists to detect smaller tumors, when curative therapies are most effective. However, the major difficulty in imaging cirrhosis is the characterization of hypervascular nodules smaller than 2 cm, which often have nonspecific imaging characteristics. The authors present a review of the MR imaging and pathologic features of regenerative nodules and dysplastic nodules and focus on HCC in the cirrhotic liver, with particular reference to small tumors and lesions that may mimic HCC. The authors also review the sensitivity of MR imaging for the detection of these tumors and discuss the staging of HCC and the treatment options in the context of the guidelines of the AASLD and the imaging criteria required by the United Network for Organ Sharing for transplantation. MR findings following ablation and chemoembolization are also reviewed. PMID- 18430872 TI - Heat distribution in the spinal canal during radiofrequency ablation for vertebral lesions: study in swine. AB - PURPOSE: To prospectively evaluate the safety of radiofrequency (RF) ablation for vertebral lesions by monitoring the temperature in swine vertebral models with and without a cortical bone defect. MATERIALS AND METHODS: The institutional animal care and use committee approved the animal studies. In vivo and ex vivo studies were performed. In the in vivo study, 20 lumbar vertebrae from six swine were locally heated by using 1- or 2-cm active-tip internally cooled electrodes. In the ex vivo study, 12 fresh pig cadaver lumbar vertebrae were extracted from four swine, and spinal tumor models with or without cortical bone defect were created by stuffing a cavity with muscle tissue and locally heated by using a 1 cm active-tip internally cooled electrode. The temperature was monitored in the spinal canal and around the vertebral body during ablation. Mann-Whitney U test was used to indicate a significant difference between groups by using 1- and 2-cm active tip in the in vivo study and between groups with and without cortical defect in the ex vivo study. RESULTS: In the in vivo study in which 1- and 2-cm active-tip needles were used, the temperature in the spinal canal rose to 38.2 degrees C +/- 2.7 (standard deviation) and 45.5 degrees C +/- 6.2, respectively. The latter was significantly higher than the former (P < .001). In the ex vivo study in which tumor models with or without a cortical bone defect were used, the temperature in the spinal canal rose to 48.4 degrees C +/- 6.2 and 31.3 degrees C +/- 3.4, respectively. The former was significantly higher than the latter (P < .001). CONCLUSION: For in vivo cases with a 2-cm active tip and ex vivo cases with a vertebral posterior bone defect, the temperature rose to over 45 degrees C, potentially injuring the spinal cord and peripheral nerves. PMID- 18430873 TI - Detection of simulated inflicted metaphyseal fractures in a fetal pig model: image optimization and dose reduction with computed radiography. AB - PURPOSE: To prospectively determine in a fetal pig model whether diagnostic performance comparable to that of high-detail screen-film imaging can be achieved with computed radiography for the detection of simulated classic metaphyseal lesions (CMLs), by using Faxitron digital images as the reference standard, and whether radiation dose reduction is possible. MATERIALS AND METHODS: This study was granted exempt status by the institutional review board and the animal care and use committee. Fractures simulating the CML were produced in distal femurs of 20 deceased fetal pigs. Twenty normal femurs served as control femurs. Femurs were imaged with a standard single-side-read 100-microm pixel sampling imaging plate (IP), a high-resolution dual-side-read 50-microm pixel sampling IP, and a high-detail screen-film imaging system. Eight tube current-time product settings (0.5-10.0 mAs) and two tube voltage selections (56 and 70 kVp) were employed. Two pediatric radiologists evaluated 920 images for fracture by using a five-point Likert scale. Area under the receiver operating characteristic curve (A(z)) values for the imaging systems were compared by using nonparametric chi(2) tests (all P < .05). RESULTS: For pooled rater data, performance of computed radiography was comparable to that of screen-film imaging, and superior performance (P = .04) was achieved with the more experienced rater. The A(z) value tended to increase as the tube current-time product setting was increased. Within each system, there was no significant difference in A(z) values for all images obtained at 56 and 70 kVp (dual-side-read IP, P = .63; single-side-read IP, P = .25; screen-film imaging system, P = .5). At 56 kVp, a dose reduction of up to 69% was achieved, and accuracy of computed radiography was comparable to that of screen-film imaging. CONCLUSION: Findings in this study suggest that computed radiography can replace screen-film imaging in the detection of CMLs and may permit dose reduction. PMID- 18430874 TI - Pulmonary nodule volume: effects of reconstruction parameters on automated measurements--a phantom study. AB - PURPOSE: To prospectively evaluate in a phantom the effects of reconstruction kernel, field of view (FOV), and section thickness on automated measurements of pulmonary nodule volume. MATERIALS AND METHODS: Spherical and lobulated pulmonary nodules 3-15 mm in diameter were placed in a commercially available lung phantom and scanned by using a 16-section computed tomographic (CT) scanner. Nodule volume (V) was determined by using the diameters of 27 spherical nodules and the mass and density values of 29 lobulated nodules measured by using the formulas V = (4/3)pi r(3) (spherical nodules) and V = 1000 x (M/D) (lobulated nodules) as reference standards, where r is nodule radius; M, nodule mass; and D, wax density. Experiments were performed to evaluate seven reconstruction kernels and the independent effects of FOV and section thickness. Automated nodule volume measurements were performed by using computer-assisted volume measurement software. General linear regression models were used to examine the independent effects of each parameter, with percentage overestimation of volume as the dependent variable of interest. RESULTS: There was no substantial difference in the accuracy of volume estimations across the seven reconstruction kernels. The bone reconstruction kernel was deemed optimal on the basis of the results of a series of statistical analyses and other qualitative findings. Overall, volume accuracy was significantly associated (P < .0001) with larger reference standard measured nodule diameter. There was substantial overestimation of the volumes of the 3-5-mm nodules measured by using the volume measurement software. Decreasing the FOV facilitated no significant improvement in the precision of lobulated nodule volume measurements. The accuracy of volume estimations--particularly those for small nodules--was significantly (P < .0001) affected by section thickness. CONCLUSION: Substantial, highly variable overestimation of volume occurs with decreasing nodule diameter. A section thickness that enables the acquisition of at least three measurements along the z-axis should be used to measure the volumes of larger pulmonary nodules. PMID- 18430875 TI - Hirschsprung disease and hypoganglionosis in adults: radiologic findings and differentiation. AB - PURPOSE: To retrospectively evaluate the imaging features of adult Hirschsprung disease (HD) and adult hypoganglionosis (HG) and to compare these features with histopathologic findings. MATERIALS AND METHODS: This study was institutional review board approved, and the requirement for informed consent was waived. The imaging, medical, and histopathologic data of 10 patients (seven women, three men; mean age, 38 years) with histopathologically proved adult HD and/or adult HG were reviewed. Two radiologists reviewed 10 transverse computed tomographic (CT) scans and five double-contrast barium enema radiographs in consensus for the presence or absence and the location of the transition zone. The transverse diameter ratio of the most dilated colonic segment proximal to the transition zone to the narrowed colonic segment distal to the transition zone (ie, transition zone ratio), and the longitudinal length of the transition zone were also determined. The CT findings of HD and HG were compared by using the Mann Whitney U test. RESULTS: All patients with lifelong or chronic constipation had a transition zone in the upper part of the rectum or rectosigmoid junction (n = 5) or in the descending colon (n = 5) on the CT scans and the double-contrast barium enema radiographs. The transition zone ratio was significantly different between the patients with HD (median ratio, 4.0) and the patients with HG (median ratio, 2.0) (P = .016). However, there was no significant difference in the longitudinal length of the transition zone between the two patient groups (median ratios, 4.4 cm for HD group and 6.0 cm for HG group; P = .190). CONCLUSION: A markedly dilated proximal colonic segment with a transition zone and a narrowed distal colonic segment on CT and double-contrast barium enema images in conjunction with chronic refractory constipation in an adult should suggest the diagnosis of adult HD or adult HG. The detection of a much higher transition zone ratio may help to establish the diagnosis of HD. PMID- 18430876 TI - Autoimmune pancreatitis: CT patterns and their changes after steroid treatment. AB - PURPOSE: To retrospectively evaluate the computed tomographic (CT) patterns of autoimmune pancreatitis (AIP) and their changes after steroid therapy. MATERIALS AND METHODS: Investigational review board approval was obtained, and the informed consent requirement was waived. The medical and imaging data of 21 patients (13 men, eight women; mean age, 47.5 years; age range, 25-79 years) with histopathologically proved AIP who underwent contrast material-enhanced CT at diagnosis and after steroid treatment were included in this study. Image analysis included assessment of the (a) presence or absence and type (focal or diffuse) of pancreatic parenchyma enlargement, (b) contrast enhancement of pancreatic parenchyma, (c) size of the main pancreatic duct (MPD) within the lesion and upstream, and (d) pancreatic parenchyma thickness in the head, body, and tail of the pancreas. The same criteria were applied to follow-up CT examinations, the follow-up data were compared with pretreatment data, and a paired sample t test was applied. RESULTS: Pancreatic parenchyma showed focal enlargement in 14 (67%) patients and diffuse enlargement in seven (33%). Pancreatic parenchyma affected by AIP appeared hypoattenuating in 19 (90%) patients and isoattenuating in two (10%). During the portal venous phase, pancreatic parenchyma showed contrast material retention in 18 (86%) patients and contrast material washout in three (14%). The MPD was never visible within the lesion. After treatment, there was a reduction in the size of pancreatic parenchyma segments affected by AIP (P < .05). Fifteen (71%) of the 21 patients had a normal enhancement pattern in the pancreatic parenchyma, whereas the enhancement pattern remained hypovascular in six (29%). The MPD returned to its normal size within the lesion in all patients at follow-up CT. In one of the eight patients with focal forms of AIP, the upstream MPD remained dilated. CONCLUSION: AIP appeared as pancreatic parenchyma enlargement, with MPD stenosis within the lesion and upstream dilatation in focal forms of AIP. After steroid treatment, there was normalization of these findings. PMID- 18430877 TI - Prostate cancer: is inapparent tumor at endorectal MR and MR spectroscopic imaging a favorable prognostic finding in patients who select active surveillance? AB - PURPOSE: To retrospectively determine whether inapparent tumor at endorectal magnetic resonance (MR) imaging and MR spectroscopic imaging is a favorable prognostic finding in prostate cancer patients who select active surveillance for management. MATERIALS AND METHODS: Committee on Human Research approval was obtained and compliance with HIPAA regulations was observed, with waiver of requirement for written consent. Ninety-two men (mean age, 64 years; range, 43-85 years) were retrospectively identified who had biopsy-proved prostate cancer, who had undergone baseline endorectal MR imaging and MR spectroscopic imaging, and who had selected active surveillance for management. Their mean baseline serum prostate-specific antigen (PSA) level was 5.5 ng/mL, and the median Gleason score was 6. Two readers with 10 and 3 years of experience independently reviewed all MR images and determined whether tumor was apparent on the basis of evaluation of established morphologic and metabolic findings. Another investigator compiled data about baseline clinical stage, biopsy findings, and serum PSA measurements. Multiple logistic regression analysis was used to investigate the relationship between the clinical parameters and tumor apparency at MR imaging and the biochemical outcome. RESULTS: At baseline MR imaging, readers 1 and 2 considered 54 and 26 patients, respectively, to have inapparent tumor (fair interobserver agreement; kappa = 0.30). During a mean follow-up of 4.8 years, 52 patients had a stable PSA level and 40 had an increasing PSA level. In multivariate analysis, no significant association was found between the baseline clinical stage, Gleason score, serum PSA level, or the presence of apparent tumor at endorectal MR imaging and MR spectroscopic imaging for either reader and the biochemical outcome (P > .05 for all). CONCLUSION: Endorectal MR imaging and MR spectroscopic imaging findings of tumor apparency or inapparency in prostate cancer patients who select active surveillance for management do not appear to be of prognostic value. PMID- 18430878 TI - Renal masses: characterization with diffusion-weighted MR imaging--a preliminary experience. AB - PURPOSE: To retrospectively assess the usefulness of apparent diffusion coefficients (ADCs) for characterizing renal masses (ie, viable solid tumors, necrotic or cystic tumor areas, and benign cysts). MATERIALS AND METHODS: The institutional review board waived the requirement for informed consent for this retrospective HIPAA-compliant study. The data of 25 consecutive patients (15 men, 10 women; age range, 39-75 years) who underwent renal magnetic resonance (MR) imaging, including diffusion-weighted imaging, before nephrectomy were included. Renal MR examinations were performed by using transverse T1-weighted dual-echo in phase and out-of-phase sequences and transverse and coronal T2-weighted single shot fast spin-echo sequences. Three-dimensional fat-saturated T1-weighted dynamic gadopentetate dimeglumine-enhanced sequences also were performed. Precontrast single-shot spin-echo echo-planar diffusion-weighted images were obtained with b values of 0, 500, and 1000 sec/mm(2) at 1.5 T. Regions of interest were placed on renal lesions to measure the ADC of whole lesions, enhancing viable soft tissue, and nonenhancing necrotic or cystic areas. The T1 signal characteristics of the renal lesions and necrotic or cystic areas were recorded. The Wilcoxon rank sum test was used to compare the median ADC values of the various types of lesions and areas. RESULTS: Twenty-six renal tumors were found in the 25 patients. Eight patients were found to have 11 benign cysts. Renal tumors had significantly lower ADCs (median, 189.3 x 10(-5) mm(2)/sec; range, [102.0-262.0] x 10(-5) mm(2)/sec) compared with benign cysts (median, 322.8 x 10(-5) mm(2)/sec; range, [217.0-421.0] x 10(-5) mm(2)/sec; P < .001). Solid enhancing tumors had significantly lower ADCs (median, 162.3 x 10(-5) mm(2)/sec; range, [102.0-284.0] x 10(-5) mm(2)/sec) compared with nonenhancing necrotic or cystic regions (median, 247.7 x 10(-5) mm(2)/sec; range, [85.2-310.0] x 10(-5) mm(2)/sec; P = .007) [corrected]. T1 hyperintense lesions had lower ADCs compared with their hypointense counterparts. CONCLUSION: The T1 signal characteristics of a renal lesion appear to be related to the ADC of the lesion. ADC may be helpful in characterizing and differentiating renal masses. PMID- 18430879 TI - Proximal femur specimens: automated 3D trabecular bone mineral density analysis at multidetector CT--correlation with biomechanical strength measurement. AB - PURPOSE: To prospectively evaluate an automated volume of interest (VOI)-fitting algorithm for quantitative computed tomography (CT) of proximal femur specimens, correlate bone mineral density (BMD) with biomechanically determined bone strength in vitro, and compare that correlation with those observed at dual energy x-ray absorptiometry (DXA) measurement of BMD. MATERIALS AND METHODS: The study was compliant with institutional and legislative requirements; donors had dedicated their body for education and research before death. Multidetector CT and DXA scans were acquired in 178 proximal femur specimens harvested from human cadavers (91 women, 87 men; mean age at death, 79 years +/- 10.2; range, 52-100 years). An automated VOI-fitting algorithm was used to calculate BMD and bone mineral content (BMC) in the head, neck, and trochanter from CT findings and pixel distribution parameters. The femur failure load (FL) was determined by using a mechanical test. Quantitative CT BMD, quantitative CT pixel distribution parameters, DXA BMD, and FL were correlated at multiple regression analysis. RESULTS: Mean precision errors in quantitative CT BMD measurements at segmentation with repositioning were 0.56%, 2.26%, and 0.61% for the head, neck, and trochanter, respectively. For the head, neck, and trochanter, respectively, r values were 0.77, 0.53, and 0.59 for the correlation between quantitative CT BMD and FL and 0.74, 0.55, and 0.65 for the correlation between quantitative CT BMC and FL (P < .001). Values ranged from 0.77 to 0.80 for correlations between DXA BMD and FL and from 0.73 to 0.82 for correlations between DXA BMC and FL (P < .001). In a multiple regression model that included quantitative CT pixel distributions, adjusted multivariate correlation coefficient values for correlations with FL increased to up to 0.88. CONCLUSION: Regional BMD of the proximal femur can be determined in vitro from quantitative CT data with high precision by using an automated VOI-fitting algorithm. The best multiple regression model for predicting FL included DXA BMD and regional quantitative CT BMD measurements. PMID- 18430880 TI - Frequency and cause of disagreements in diagnoses for fetuses referred for ventriculomegaly. AB - PURPOSE: To prospectively assess the frequency and cause of disagreements in diagnoses at ultrasonography (US) and magnetic resonance (MR) imaging for fetuses referred for ventriculomegaly (VM). MATERIALS AND METHODS: One hundred ninety five women, aged 18-44 years, with 200 fetal referrals for VM, were recruited in a prospective IRB-approved, HIPAA-compliant study. Written informed consent was obtained. US scans were prospectively interpreted by three obstetric radiologists and MR examinations were read by one obstetric radiologist and three pediatric neuroradiologists. Final diagnosis was reached by consensus (198 US, 198 MR, and 196 US-MR comparisons). Gestational age, ventricular size, types of disagreements, and reasons for disagreements were recorded. Interreader agreement was assessed with kappa statistics. Ventricular diameter, gestational age, and confidence scores were analyzed by using mixed-model analysis of variance, accounting for correlation within reader and fetus. RESULTS: There was prospective agreement on 118 (60%) of 198 US and 104 (53%) of 198 MR readings. Consensus was more likely when the final diagnosis was isolated VM (83 of 104, 80% at US; 82 of 109, 75% at MR) than when the final diagnosis included other anomalies as well (14 of 63, 22% at US; seven of 68, 10% at MR; P < .001). There was disagreement on 19 (10%) of 196 and 31 (16%) of 196 fetuses about the presence of VM at US and MR, respectively, and on 29 (15%) of 198 and 39 (20%) of 198 fetuses regarding the presence of major findings at US and MR, respectively. Reasons for discrepancies in reporting major findings included errors of observation, lack of real-time US scanning, lack of neuroradiology experience, as well as modality differences in helping depict abnormalities. CONCLUSION: Of radiologists who read high-risk obstetric US and fetal MR images for VM, there is considerable variability in central nervous system diagnosis. PMID- 18430881 TI - Is propofol a safe alternative to pentobarbital for sedation during pediatric diagnostic CT? AB - PURPOSE: To prospectively compare the incidence of adverse respiratory events, the need for airway interventions, and the recovery time after propofol sedation with similar data from a retrospective review of data obtained in patients who underwent pentobarbital sedation. MATERIALS AND METHODS: This HIPAA-compliant study was conducted with institutional review board approval and parental informed consent. The hospital sedation committee approved a 2-month pilot program of propofol sedation as a potential alternative to pentobarbital sedation. Parents were given the choice of having their child sedated with intravenously administered propofol or pentobarbital. Fifty-two patients (18 female, 34 male; mean age, 2.9 years +/- 2.4 [standard deviation]) received propofol. An equal number of patients (21 female, 31 male; mean age, 2.5 years +/ 1.7) who previously received pentobarbital were included. The sample sizes provided 80% power to detect differences in airway manipulations, adverse respiratory events, and recovery time between the groups by using the Fisher exact test and the Student t test. A two-tailed P value of less than .05 indicated a significant difference. RESULTS: Patients sedated with propofol underwent significantly more airway manipulations to relieve obstruction than did patients sedated with pentobarbital (23% vs 0%, P < .001). More adverse respiratory events occurred in the propofol group than in the pentobarbital group (12% vs 0%, P = .03). Patients in the propofol group had a faster recovery profile than did patients in the pentobarbital group (34 minutes +/- 17 vs 100 minutes +/- 30, P < .001). CONCLUSION: Propofol is associated with a significantly greater incidence of adverse respiratory events than is pentobarbital. PMID- 18430882 TI - Transperineal prostate intervention: robot for fully automated MR imaging--system description and proof of principle in a canine model. AB - The study was approved by the animal care and use committee. The purpose of the study was to prospectively establish proof of principle in vivo in canines for a magnetic resonance (MR) imaging-compatible robotic system designed for image guided prostatic needle intervention. The entire robot is built with nonmagnetic and dielectric materials and in its current configuration is designed to perform fully automated brachytherapy seed placement within a closed MR imager. With a 3.0-T imager, in four dogs the median error for MR imaging-guided needle positioning and seed positioning was 2.02 mm (range, 0.86-3.18 mm) and 2.50 mm (range, 1.45-10.54 mm), respectively. The robotic system is capable of accurate MR imaging-guided prostatic needle intervention within a standard MR imager in vivo in a canine model. PMID- 18430884 TI - Case 133: Ecchordosis physaliphora. PMID- 18430885 TI - The incomplete fissure sign. PMID- 18430886 TI - MR elastography to assess liver fibrosis. PMID- 18430887 TI - Same-specialty referrals for imaging compared with referrals to radiologists: are the data correct? PMID- 18430888 TI - On radiology referral versus specialist referral. PMID- 18430890 TI - Synthesis and characterization of mRNA cap analogs containing phosphorothioate substitutions that bind tightly to eIF4E and are resistant to the decapping pyrophosphatase DcpS. AB - Analogs of the mRNA cap are widely employed to study processes involved in mRNA metabolism as well as being useful in biotechnology and medicinal applications. Here we describe synthesis of six dinucleotide cap analogs bearing a single phosphorothioate modification at either the alpha, beta, or gamma position of the 5',5'-triphosphate chain. Three of them were also modified with methyl groups at the 2'-O position of 7-methylguanosine to produce anti-reverse cap analogs (ARCAs). Due to the presence of stereogenic P centers in the phosphorothioate moieties, each analog was obtained as a mixture of two diastereomers, D1 and D2. The mixtures were resolved by RP HPLC, providing 12 different compounds. Fluorescence quenching experiments were employed to determine the association constant (K(AS)) for complexes of the new analogs with eIF4E. We found that phosphorothioate modifications generally stabilized the complex between eIF4E and the cap analog. The most strongly bound phosphorothioate analog (the D1 isomer of the beta-substituted analog m(7)Gpp(S)pG) was characterized by a K(AS) that was more than fourfold higher than that of its unmodified counterpart (m(7)GpppG). All analogs modified in the gamma position were resistant to hydrolysis by the scavenger decapping pyrophosphatase DcpS from both human and Caenorhabditis elegans sources. The absolute configurations of the diastereomers D1 and D2 of analogs modified at the alpha position (i.e., m(7)Gppp(S)G and m(2) (7,2'-O )Gppp(S)G) were established as S(P) and R(P) , respectively, using enzymatic digestion and correlation with the S(P) and R(P) diastereomers of guanosine 5'-O (1-thiodiphosphate) (GDPalphaS). The analogs resistant to DcpS act as potent inhibitors of in vitro protein synthesis in rabbit reticulocyte lysates. PMID- 18430891 TI - A multifunctional human Argonaute2-specific monoclonal antibody. AB - Small regulatory RNAs including small interfering RNAs (siRNAs), microRNAs (miRNAs), or Piwi interacting RNAs (piRNAs) guide regulation of gene expression in many different organisms. The Argonaute (Ago) protein family constitutes the cellular binding partners of such small RNAs and regulates gene expression on the levels of transcription, mRNA stability, or translation. Due to the lack of highly specific and potent monoclonal antibodies directed against the different Ago proteins, biochemical analyses such as Ago complex purification and characterization rely on overexpression of tagged Ago proteins. Here, we report the generation and functional characterization of a highly specific monoclonal anti-Ago2 antibody termed anti-Ago2(11A9). We show that anti-Ago2(11A9) is specific for human Ago2 and detects Ago2 in Western blots as well as in immunoprecipitation experiments. We further demonstrate that Ago2 can be efficiently eluted from our antibody by a competing peptide. Finally, we show that anti-Ago2(11A9) recognizes Ago2 in immunofluorescence experiments, and we find that Ago2 not only localizes to cytoplasmic processing bodies (P-bodies) and the diffuse cytoplasm but also to the nucleus. With the anti-Ago2(11A9) antibody we have generated a potent tool that is useful for many biochemical or cell biological applications. PMID- 18430892 TI - Specificity of ADAR-mediated RNA editing in newly identified targets. AB - Adenosine deaminases that act on RNA (ADARs) convert adenosines to inosine in both coding and noncoding double-stranded RNA. Deficiency in either ADAR1 or ADAR2 in mice is incompatible with normal life and development. While the ADAR2 knockout phenotype can be attributed to the lack of editing of the GluR-B receptor, the embryonic lethal phenotype caused by ADAR1 deficiency still awaits clarification. Recently, massive editing was observed in noncoding regions of mRNAs in mice and humans. Moreover, editing was observed in protein-coding regions of four mRNAs encoding FlnA, CyFip2, Blcap, and IGFBP7. Here, we investigate which of the two active mammalian ADAR enzymes is responsible for editing of these RNAs and whether any of them could possibly contribute to the phenotype observed in ADAR knockout mice. Editing of Blcap, FlnA, and some sites within B1 and B2 SINEs clearly depends on ADAR1, while other sites depend on ADAR2. Based on our data, substrate specificities can be further defined for ADAR1 and ADAR2. Future studies on the biological implications associated with a changed editing status of the studied ADAR targets will tell whether one of them turns out to be directly or indirectly responsible for the severe phenotype caused by ADAR1 deficiency. PMID- 18430894 TI - Effects of the Aurora kinase inhibitor VX-680 on anaplastic thyroid cancer derived cell lines. AB - Anaplastic thyroid cancers (ATC) are aggressive tumors, which exhibit cell cycle misregulations leading to uncontrolled cellular proliferation and genomic instability. They fail to respond to chemotherapeutic agents and radiation therapy, and most patients die within a few months of diagnosis. In the present study, we evaluated the in vitro effects on ATC cells of VX-680, an inhibitor of the Aurora serine/threonine kinases involved in the regulation of multiple aspects of chromosome segregation and cytokinesis. The effects of VX-680 on proliferation, apoptosis, soft agar colony formation, cell cycle, and ploidy were tested on the ATC-derived cell lines CAL-62, 8305C, 8505C, and BHT-101. Treatment of the different ATC cells with VX-680 inhibited proliferation in a time- and dose-dependent manner, with the IC50 between 25 and 150 nM. The VX-680 significantly impaired the ability of the different cell lines to form colonies in soft agar. Analysis of caspase-3 activity showed that VX-680 induced apoptosis in the different cell lines. CAL-62 cells exposed for 12 h to VX-680 showed an accumulation of cells with > or =4N DNA content. Time-lapse analysis demonstrated that VX-680-treated CAL-62 cells exit metaphase without dividing. Moreover, histone H3 phosphorylation was abrogated following VX-680 treatment. In conclusion, our data demonstrated that VX-680 is effective in reducing cell growth of different ATC-derived cell lines and warrant further investigation to exploit its potential therapeutic value for ATC treatment. PMID- 18430893 TI - Riboswitch effectors as protein enzyme cofactors. AB - The recently identified glmS ribozyme revealed that RNA enzymes, like protein enzymes, are capable of using small molecules as catalytic cofactors to promote chemical reactions. Flavin mononucleotide (FMN), S-adenosyl methionine (SAM), adenosyl cobalamin (AdoCbl), and thiamine pyrophosphate (TPP) are known ligands for RNA riboswitches in the control of gene expression, but are also catalytically powerful and ubiquitous cofactors in protein enzymes. If RNA, instead of just binding these molecules, could harness the chemical potential of the cofactor, it would significantly expand the enzymatic repertoire of ribozymes. Here we review the chemistry of AdoCbl, SAM, FMN, and TPP in protein enzymology and speculate on how these cofactors might have been used by ribozymes in the prebiotic RNA World or may still find application in modern biology. PMID- 18430895 TI - Analysis of single nucleotide polymorphisms of FSH receptor gene suggests association with testicular cancer susceptibility. AB - The development of testicular germ cell tumour (TGCT) is believed to be under endocrine control but definitive proofs are lacking. Follicle stimulating hormone (FSH) levels are increased in numerous conditions associated with increased risk of TGCT and single nucleotide polymorphisms (SNPs) in the FSH receptor (FSHR) gene influence the sensitivity of the receptor to FSH. However, a possible effect of FSH on testicular carcinogenesis has never been explored. In order to analyse the possible association of FSHR polymorphisms with TGCT, we studied 188 TGTC cases and 152 controls for 12 FSHR SNPs. Only four SNPs were found to be informative, represented by two polymorphisms in exon 10 (Ala307Thr and Ser680Asn), and two polymorphisms in the promoter region (-114 T/C and -29 G/A). Differences in haplotype distribution were seen between TGCT cases and controls. In particular for non-seminoma, the Ala307/Ser680 allele lowers the risk of the disease, alone (P=0.014, relative risk 0.73; 95% confidence interval 0.57-0.92), or in combination with the -29 G allele and/or the -114 T allele. This study suggests for the first time that FSHR gene polymorphisms modulate susceptibility to TGCT. The variants with higher activity of the FSHR are associated with higher risk, suggesting a role for FSH in the carcinogenesis of this tumour. PMID- 18430896 TI - Regulation of tumor necrosis factor-related apoptosis-inducing ligand-induced apoptosis by DJ-1 in thyroid cancer cells. AB - DJ-1, a cancer-associated protein protects cells from multiple toxic stresses. The expression of DJ-1 and its influence on thyroid cancer cell death has not been investigated so far. We analyzed DJ-1 expression in human thyroid carcinoma cell lines and the effect of DJ-1 on tumor necrosis factor-related apoptosis inducing ligand (TRAIL)-induced apoptosis. DJ-1 was expressed in human thyroid carcinoma cell lines; small interfering RNA-mediated downregulation of its levels significantly sensitized thyroid carcinoma cells to TRAIL-induced apoptosis, whereas the forced exogenous expression of DJ-1 significantly suppressed cell death induced by TRAIL. We also report here that TRAIL-induced thyroid cancer cell apoptosis is mediated by oxidative stress and that DJ-1, a potent nutritional antioxidant, protects cancer cells from apoptosis at least in part by impeding the elevation of reactive oxygen species levels induced by TRAIL and impairing caspase-8 activation. Subsequently, we investigated DJ-1 expression in 52 normal and 74 primary thyroid carcinomas from patients of China Medical University. The protein was not detectable in the 52 specimens of normal thyroid, while 70 out of 74 analyzed carcinomas (33 out of 33 follicular, 17 out of 19 papillary, 12 out of 13 medullar, and 8 out of 9 anaplastic) were clearly positive for DJ-1 expression. Our data demonstrated that DJ-1 is specifically expressed in thyroid carcinomas and not in the normal thyroid tissue. In addition, the protein modulates the response to TRAIL-mediated apoptosis in human neoplastic thyroid cells, at least partially through its antioxidant property. PMID- 18430897 TI - Amyloid precursor-like protein 1 is differentially upregulated in neuroendocrine tumours of the gastrointestinal tract. AB - We have examined the global gene expression profile of small intestinal carcinoids by microarray analysis. High expression of a number of genes was found including amyloid precursor-like protein 1 (APLP1). Quantitative real-time PCR and western blot analysis demonstrated higher expression of APLP1 in carcinoid metastases relative to primary tumours indicating a role of APLP1 in tumour dissemination. Tissue microarray analysis of gastroentero-pancreatic tumours demonstrated a high frequency of APLP1 expression and a low frequency of APLP2 expression in neuroendocrine (NE) tumours when compared with non-NE tumours at the same sites. Meta-analysis of gene expression data from a large number of tumours outside the gastrointestinal tract confirmed a correlation between APLP1 expression and NE phenotype where high expression of APLP1 was accompanied by downregulation of APLP2 in NE tumours. Cellular localization of APLP1, APLP2 and amyloid precursor protein (APP) in carcinoid cells (GOT1) by confocal microscopy demonstrated partial co-localization with synaptophysin. This suggests that the APP family of proteins is transported to the cell membrane by synaptic microvesicles and that they may influence tumour cell adhesion and invasiveness. We conclude that APLP1 is differentially upregulated in gastrointestinal NE tumours and that APLP1 may be important for the dissemination of small intestinal carcinoids. Identification of APLP1 in NE tumours offers a novel target for treatment and may also serve as a tumour-specific marker. PMID- 18430898 TI - The cover. Portrait of Marthe Marie Tronchin. PMID- 18430899 TI - A piece of my mind. Thank goodness for my ass. PMID- 18430900 TI - VA, military seek office-based PTSD care. PMID- 18430901 TI - Scientists examine primary care-based screening and treatments for anxiety. PMID- 18430902 TI - Researchers seek ways to stem STDs: "alarming" STD rates found in teenaged girls. PMID- 18430903 TI - Guidelines for von Willebrand disease urge routine screening for bleeding. PMID- 18430904 TI - Misinterpretation of electrocardiograms and cardiac catheterization laboratory activations. PMID- 18430905 TI - Antipsychotic-induced weight gain in patients with schizophrenia. PMID- 18430906 TI - Antipsychotic-induced weight gain in patients with schizophrenia. PMID- 18430907 TI - Effects of testosterone therapy in older men. PMID- 18430908 TI - Applying market justice to health care. PMID- 18430910 TI - Fluorouracil, mitomycin, and radiotherapy vs fluorouracil, cisplatin, and radiotherapy for carcinoma of the anal canal: a randomized controlled trial. AB - CONTEXT: Chemoradiation as definitive therapy is the preferred primary therapy for patients with anal canal carcinoma; however, the 5-year disease-free survival rate from concurrent fluorouracil/mitomycin and radiation is only approximately 65%. OBJECTIVE: To compare the efficacy of cisplatin-based (experimental) therapy vs mitomycin-based (standard) therapy in treatment of anal canal carcinoma. DESIGN, SETTING, AND PARTICIPANTS: US Gastrointestinal Intergroup trial RTOG 98 11, a multicenter, phase 3, randomized controlled trial comparing treatment with fluorouracil plus mitomycin and radiotherapy vs treatment with fluorouracil plus cisplatin and radiotherapy in 682 patients with anal canal carcinoma enrolled between October 31, 1998, and June 27, 2005. Stratifications included sex, clinical nodal status, and tumor diameter. INTERVENTION: Participants were randomly assigned to 1 of 2 intervention groups: (1) the mitomycin-based group (n = 341), who received fluorouracil (1000 mg/m2 on days 1-4 and 29-32) plus mitomycin (10 mg/m2 on days 1 and 29) and radiotherapy (45-59 Gy) or (2) the cisplatin-based group (n = 341), who received fluorouracil (1000 mg/m2 on days 1 4, 29-32, 57-60, and 85-88) plus cisplatin (75 mg/m2 on days 1, 29, 57, and 85) and radiotherapy (45-59 Gy; start day = day 57). MAIN OUTCOME MEASURES: The primary end point was 5-year disease-free survival; secondary end points were overall survival and time to relapse. RESULTS: A total of 644 patients were assessable. The median follow-up for all patients was 2.51 years. Median age was 55 years, 69% were women, 27% had a tumor diameter greater than 5 cm, and 26% had clinically positive nodes. The 5-year disease-free survival rate was 60% (95% confidence interval [CI], 53%-67%) in the mitomycin-based group and 54% (95% CI, 46%-60%) in the cisplatin-based group (P = .17). The 5-year overall survival rate was 75% (95% CI, 67%-81%) in the mitomycin-based group and 70% (95% CI, 63%-76%) in the cisplatin-based group (P = .10). The 5-year local-regional recurrence and distant metastasis rates were 25% (95% CI, 20%-30%) and 15% (95% CI, 10%-20%), respectively, for mitomycin-based treatment and 33% (95% CI, 27%-40%) and 19% (95% CI, 14%-24%), respectively, for cisplatin-based treatment. The cumulative rate of colostomy was significantly better for mitomycin-based than cisplatin based treatment (10% vs 19%; P = .02). Severe hematologic toxicity was worse with mitomycin-based treatment (P < .001). CONCLUSIONS: In this population of patients with anal canal carcinoma, cisplatin-based therapy failed to improve disease-free survival compared with mitomycin-based therapy, but cisplatin-based therapy resulted in a significantly worse colostomy rate. These findings do not support the use of cisplatin in place of mitomycin in combination with fluorouracil and radiotherapy in the treatment of anal canal carcinoma. TRIAL REGISTRATION: clinicaltrials.gov Identifier: NCT00003596. PMID- 18430911 TI - Cost-related medication nonadherence and spending on basic needs following implementation of Medicare Part D. AB - CONTEXT: Cost-related medication nonadherence (CRN) has been a persistent problem for individuals who are elderly and disabled in the United States. The impact of Medicare prescription drug coverage (Part D) on CRN is unknown. OBJECTIVE: To estimate changes in CRN and forgoing basic needs to pay for drugs following Part D implementation. DESIGN, SETTING, AND PARTICIPANTS: In a population-level study design, changes in study outcomes between 2005 and 2006 before and after Medicare Part D implementation were compared with historical changes between 2004 and 2005. The community-dwelling sample of the nationally representative Medicare Current Beneficiary Survey (unweighted unique n = 24,234; response rate, 72.3%) was used, and logistic regression analyses were controlled for demographic characteristics, health status, and historical trends. MAIN OUTCOME MEASURES: Self-reports of CRN (skipping or reducing doses, not obtaining prescriptions) and spending less on basic needs to afford medicines. RESULTS: The unadjusted, weighted prevalence of CRN was 15.2% in 2004, 14.1% in 2005, and 11.5% after Part D implementation in 2006. The prevalence of spending less on basic needs was 10.6% in 2004, 11.1% in 2005, and 7.6% in 2006. Adjusted analyses comparing 2006 with 2005 and controlling for historical changes (2005 vs 2004) demonstrated significant decreases in the odds of CRN (ratio of odds ratios [ORs], 0.85; 95% confidence interval [CI], 0.74-0.98; P = .03) and spending less on basic needs (ratio of ORs, 0.59; 95% CI, 0.48-0.72; P < .001). No significant changes in CRN were observed among beneficiaries with fair to poor health (ratio of ORs, 1.00; 95% CI, 0.82-1.21; P = .97), despite high baseline CRN prevalence for this group (22.2% in 2005) and significant decreases among beneficiaries with good to excellent health (ratio of ORs, 0.77; 95% CI, 0.63-0.95; P = .02). However, significant reductions in spending less on basic needs were observed in both groups (fair to poor health: ratio of ORs, 0.60; 95% CI, 0.47-0.75; P < .001; and good to excellent health: ratio of ORs, 0.57; 95% CI, 0.44-0.75; P < .001). CONCLUSIONS: In this survey population, there was evidence for a small but significant overall decrease in CRN and forgoing basic needs following Part D implementation. However, no net decrease in CRN after Part D was observed among the sickest beneficiaries, who continued to experience higher rates of CRN. PMID- 18430909 TI - Comparison of an everolimus-eluting stent and a paclitaxel-eluting stent in patients with coronary artery disease: a randomized trial. AB - CONTEXT: A thin, cobalt-chromium stent eluting the antiproliferative agent everolimus from a nonadhesive, durable fluoropolymer has shown promise in preliminary studies in improving clinical and angiographic outcomes in patients with coronary artery disease. OBJECTIVE: To evaluate the safety and efficacy of an everolimus-eluting stent compared with a widely used paclitaxel-eluting stent. DESIGN, SETTING, AND PATIENTS: The SPIRIT III trial, a prospective, randomized, single-blind, controlled trial enrolling patients at 65 academic and community based US institutions between June 22, 2005, and March 15, 2006. Patients were 1002 men and women undergoing percutaneous coronary intervention in lesions 28 mm or less in length and with reference vessel diameter between 2.5 and 3.75 mm. Angiographic follow-up was prespecified at 8 months in 564 patients and completed in 436 patients. Clinical follow-up was performed at 1, 6, 9, and 12 months. INTERVENTIONS: Patients were randomized 2:1 to receive the everolimus-eluting stent (n = 669) or the paclitaxel-eluting stent (n = 333). MAIN OUTCOME MEASURES: The primary end point was noninferiority or superiority of angiographic in segment late loss. The major secondary end point was noninferiority assessment of target vessel failure events (cardiac death, myocardial infarction, or target vessel revascularization) at 9 months. An additional secondary end point was evaluation of major adverse cardiac events (cardiac death, myocardial infarction, or target lesion revascularization) at 9 and 12 months. RESULTS: Angiographic in segment late loss was significantly less in the everolimus-eluting stent group compared with the paclitaxel group (mean, 0.14 [SD, 0.41] mm vs 0.28 [SD, 0.48] mm; difference, -0.14 [95% CI, -0.23 to -0.05]; P < or = .004). The everolimus stent was noninferior to the paclitaxel stent for target vessel failure at 9 months (7.2% vs 9.0%, respectively; difference, -1.9% [95% CI, -5.6% to 1.8%]; relative risk, 0.79 [95% CI, 0.51 to 1.23]; P < .001). The everolimus stent compared with the paclitaxel stent resulted in significant reductions in composite major adverse cardiac events both at 9 months (4.6% vs 8.1%; relative risk, 0.56 [95% CI, 0.34 to 0.94]; P = .03) and at 1 year (6.0% vs 10.3%; relative risk, 0.58 [95% CI, 0.37 to 0.90]; P = .02), due to fewer myocardial infarctions and target lesion revascularization procedures. CONCLUSIONS: In this large-scale, prospective randomized trial, an everolimus-eluting stent compared with a paclitaxel-eluting stent resulted in reduced angiographic late loss, noninferior rates of target vessel failure, and fewer major adverse cardiac events during 1 year of follow-up. TRIAL REGISTRATION: clinicaltrials.gov Identifier: NCT00180479. PMID- 18430912 TI - Medicare beneficiaries' knowledge of Part D prescription drug program benefits and responses to drug costs. AB - CONTEXT: Medicare Part D drug benefits include substantial cost sharing. OBJECTIVE: To determine beneficiaries' knowledge of benefits and cost responses. DESIGN, SETTING, AND PARTICIPANTS: Telephone interviews were conducted in 2007 in a stratified random sample of community-dwelling Kaiser Permanente-Northern California Medicare Advantage beneficiaries aged 65 years or older, with a gap in coverage if they exceeded $2250 in drug costs (N = 1040; 74.9% response rate). Half were selected to have reached the gap in 2006. In the source population of Medicare Advantage Prescription Drug plan beneficiaries, 8% entered the coverage gap in 2006. Models were adjusted for individual characteristics and weighted for sampling proportions. MAIN OUTCOME MEASURES: Knowledge of cost sharing including awareness of the coverage gap, gap start and end amounts, and drug cost sharing before, during, and after the gap. Cost-related responses including cost-coping behaviors (eg, switching to lower-cost medications), reduced adherence (eg, not refilling prescriptions), and financial burden (eg, going without necessities). RESULTS: An estimated 40% (95% confidence interval [CI], 35%-45%) of beneficiaries were aware that their drug plan in 2006 included a coverage gap; knowledge of the gap was greater among individuals who reached the gap during the year. Approximately 36% (95% CI, 32%-41%) of beneficiaries reported at least 1 of the following responses to drug costs: cost-coping behavior (26%), reduced adherence (15%), or experiencing financial burden (7%). In multivariate analyses, beneficiaries with lower household income more frequently reported cost responses (difference of 14.5 percentage points for < $40,000/y vs > or = $40,000/y [95% CI, 3.6-25.4 percentage points]). Compared with beneficiaries who were unaware of having a coverage gap, those who were aware more frequently reported any cost response (difference of 11.3 percentage points [95% CI, 0.8-21.9 percentage points]), but had fewer reports of borrowing money or going without necessities (difference of 5.5 percentage points [95% CI, 1.1-10.0 percentage points]). CONCLUSIONS: Beneficiaries in this Medicare Advantage plan have limited knowledge of Part D cost sharing and often report behavioral responses to drug costs. Limited knowledge is associated with fewer reports of cost responses overall, but more reports of financial burden. PMID- 18430913 TI - Efficacy of current drugs against soil-transmitted helminth infections: systematic review and meta-analysis. AB - CONTEXT: More than a quarter of the human population is likely infected with soil transmitted helminths (Ascaris lumbricoides, hookworm, and Trichuris trichiura) in highly endemic areas. Preventive chemotherapy is the mainstay of control, but only 4 drugs are available: albendazole, mebendazole, levamisole, and pyrantel pamoate. OBJECTIVE: To assess the efficacy of single-dose oral albendazole, mebendazole, levamisole, and pyrantel pamoate against A lumbricoides, hookworm, and T trichiura infections. DATA SOURCES: A systematic search of PubMed, ISI Web of Science, ScienceDirect, the World Health Organization library database, and the Cochrane Central Register of Controlled Trials (1960 to August 2007). STUDY SELECTION: From 168 studies, 20 randomized controlled trials were included. DATA EXTRACTION AND DATA SYNTHESIS: Information on study year and country, sample size, age of study population, mean infection intensity before treatment, diagnostic method used, time between evaluations before and after treatment, cure rate (the percentage of individuals who became helminth egg negative following treatment with an anthelminthic drug), egg reduction rate, adverse events, and trial quality was extracted. Relative risk, including a 95% confidence interval (CI), was used to measure the effect of the drugs on the risk of infection prevalence with a random-effects model. RESULTS: Single-dose oral albendazole, mebendazole, and pyrantel pamoate for infection with A lumbricoides resulted in cure rates of 88% (95% CI, 79%-93%; 557 patients), 95% (95% CI, 91%-97%; 309 patients), and 88% (95% CI, 79%-93%; 131 patients), respectively. Cure rates for infection with T trichiura following treatment with single-dose oral albendazole and mebendazole were 28% (95% CI, 13%-39%; 735 patients) and 36% (95% CI, 16% 51%; 685 patients), respectively. The efficacy of single-dose oral albendazole, mebendazole, and pyrantel pamoate against hookworm infections was 72% (95% CI, 59%-81%; 742 patients), 15% (95% CI, 1%-27%; 853 patients), and 31% (95% CI, 19% 42%; 152 patients), respectively. No pooled relative risks could be calculated for pyrantel pamoate against T trichiura and levamisole for any of the parasites investigated. CONCLUSIONS: Single-dose oral albendazole, mebendazole, and pyrantel pamoate show high cure rates against A lumbricoides. For hookworm infection, albendazole was more efficacious than mebendazole and pyrantel pamoate. Treatment of T trichiura with single oral doses of current anthelminthics is unsatisfactory. New anthelminthics are urgently needed. PMID- 18430914 TI - FDA guidance on off-label promotion and the state of the literature from sponsors. PMID- 18430915 TI - Next-generation drug-eluting stents: a spirited step forward or more of the same. PMID- 18430916 TI - Medicare Part D: a successful start with room for improvement. PMID- 18430917 TI - JAMA patient page. Anal cancer. PMID- 18430919 TI - Paramutation: epigenetic instructions passed across generations. PMID- 18430920 TI - RNAi of met1 reduces DNA methylation and induces genome-specific changes in gene expression and centromeric small RNA accumulation in Arabidopsis allopolyploids. AB - Changes in genome structure and gene expression have been documented in both resynthesized and natural allopolyploids that contain two or more divergent genomes. The underlying mechanisms for rapid and stochastic changes in gene expression are unknown. Arabidopsis suecica is a natural allotetraploid derived from the extant A. thaliana and A. arenosa genomes that are homeologous in the allotetraploid. Here we report that RNAi of met1 reduced DNA methylation and altered the expression of approximately 200 genes, many of which encode transposons, predicted proteins, and centromeric and heterochromatic RNAs. Reduced DNA methylation occurred frequently in promoter regions of the upregulated genes, and an En/Spm-like transposon was reactivated in met1-RNAi A. suecica lines. Derepression of transposons, heterochromatic repeats, and centromeric small RNAs was primarily derived from the A. thaliana genome, and A. arenosa homeologous loci were less affected by methylation defects. A high level of A. thaliana centromeric small RNA accumulation was correlated with hypermethylation of A. thaliana centromeres. The greater effects of reduced DNA methylation on transposons and centromeric repeats in A. thaliana than in A. arenosa are consistent with the repression of many genes that are expressed at higher levels in A. thaliana than in A. arenosa in the resynthesized allotetraploids. Moreover, non-CG (CC) methylation in the promoter region of A. thaliana At2g23810 remained in the resynthesized allotetraploids, and the methylation spread within the promoter region in natural A. suecica, leading to silencing of At2g23810. At2g23810 was demethylated and reactivated in met1-RNAi A. suecica lines. We suggest that many A. thaliana genes are transcriptionally repressed in resynthesized allotetraploids, and a subset of A. thaliana loci including transposons and centromeric repeats are heavily methylated and subjected to homeologous genome-specific RNA-mediated DNA methylation in natural allopolyploids. PMID- 18430921 TI - A Mutator transposon insertion is associated with ectopic expression of a tandemly repeated multicopy Myb gene pericarp color1 of maize. AB - The molecular basis of tissue-specific pigmentation of maize carrying a tandemly repeated multicopy allele of pericarp color1 (p1) was examined using Mutator (Mu) transposon-mediated mutagenesis. The P1-wr allele conditions a white or colorless pericarp and a red cob glumes phenotype. However, a Mu-insertion allele, designated as P1-wr-mum6, displayed an altered phenotype that was first noted as occasional red stripes on pericarp tissue. This gain-of-pericarp-pigmentation phenotype was heritable, yielding families that displayed variable penetrance and expressivity. In one fully penetrant family, deep red pericarp pigmentation was observed. Several reports on Mu suppressible alleles have shown that Mu transposons can affect gene expression by mechanisms that depend on transposase activity. Conversely, the P1-wr-mum6 phenotype is not affected by transposase activity. The increased pigmentation was associated with elevated mRNA expression of P1-wr-mum6 copy (or copies) that was uninterrupted by the transposons. Genomic bisulfite sequencing analysis showed that the elevated expression was associated with hypomethylation of a floral-specific enhancer that is approximately 4.7 kb upstream of the Mu1 insertion site and may be proximal to an adjacent repeated copy. We propose that the Mu1 insertion interferes with the DNA methylation and related chromatin packaging of P1-wr, thereby inducing expression from gene copy (or copies) that is otherwise suppressed. PMID- 18430922 TI - The Caenorhabditis elegans rsd-2 and rsd-6 genes are required for chromosome functions during exposure to unfavorable environments. AB - In Caenorhabditis elegans, exogenous dsRNA can elicit systemic RNAi, a process that requires the function of many genes. Considering that the activities of many of these genes are also required for normal development, it is surprising that exposure to high concentrations of dsRNA does not elicit adverse consequences to animals. Here, we report inducible phenotypes in attenuated C. elegans strains reared in environments that include nonspecific dsRNA and elevated temperature. Under these conditions, chromosome integrity is compromised in RNAi-defective strains harboring mutations in rsd-2 or rsd-6. Specifically, rsd-2 mutants display defects in transposon silencing, while meiotic chromosome disjunction is affected in rsd-6 mutants. RSD-2 proteins localize to multiple cellular compartments, including the nucleolus and cytoplasmic compartments that, in part, are congruent with calreticulin and HAF-6. We considered that the RNAi defects in rsd-2 mutants might have relevance to membrane-associated functions; however, endomembrane compartmentalization and endocytosis/exocytosis markers in rsd-2 and rsd-6 mutants appear normal. The mutants also possess environmentally sensitive defects in cell-autonomous RNAi elicited from transgene-delivered dsRNAs. Thus, the ultimate functions of rsd-2 and rsd-6 in systemic RNAi are remarkably complex and environmentally responsive. PMID- 18430923 TI - Genetic dissociation of ethanol sensitivity and memory formation in Drosophila melanogaster. AB - The ad hoc genetic correlation between ethanol sensitivity and learning mechanisms in Drosophila could overemphasize a common process supporting both behaviors. To challenge directly the hypothesis that these mechanisms are singular, we examined the learning phenotypes of 10 new strains. Five of these have increased ethanol sensitivity, and the other 5 do not. We tested place and olfactory memory in each of these lines and found two new learning mutations. In one case, altering the tribbles gene, flies have a significantly reduced place memory, elevated olfactory memory, and normal ethanol response. In the second case, mutation of a gene we name ethanol sensitive with low memory (elm), place memory was not altered, olfactory memory was sharply reduced, and sensitivity to ethanol was increased. In sum, however, we found no overall correlation between ethanol sensitivity and place memory in the 10 lines tested. Furthermore, there was a weak but nonsignificant correlation between ethanol sensitivity and olfactory learning. Thus, mutations that alter learning and sensitivity to ethanol can occur independently of each other and this implies that the set of genes important for both ethanol sensitivity and learning is likely a subset of the genes important for either process. PMID- 18430924 TI - The Saccharomyces cerevisiae actin cytoskeletal component Bsp1p has an auxiliary role in actomyosin ring function and in the maintenance of bud-neck structure. AB - Iqg1p is a component of the actomyosin contractile ring that is required for actin recruitment and septum deposition. Cells lacking Iqg1p function have an altered bud-neck structure and fail to form a functional actomyosin contractile ring resulting in a block to cytokinesis and septation. Here it is demonstrated that increased expression of the actin cytoskeleton associated protein Bsp1p bypasses the requirement for contractile ring function. This also correlates with reduced bud-neck width and remedial septum formation. Increased expression of this protein in a temperature-sensitive iqg1-1 background causes remedial septum formation at the bud neck that is reliant upon chitin synthase III activity and restores cell separation. The observed suppression correlates with a restoration of normal bud-neck structure. While Bsp1p is a component of the contractile ring, its recruitment to the bud neck is not required for the observed suppression. Loss of Bsp1p causes a brief delay in the redistribution of the actin cytoskeleton normally observed at the end of actin ring contraction. Compromise of Iqg1p function, in the absence of Bsp1p function, leads to a profound change in the distribution of actin and the pattern of cell growth accompanied by a failure to complete cytokinesis and cell separation. PMID- 18430925 TI - Epigenetic control may explain large within-plant heterogeneity of meiotic behavior in telocentric trisomics of rye. AB - In telocentric trisomics (telotrisomics) of organisms in which the chromosomes normally have two distinct arms, a single chromosome arm with a centromere is present in addition to a complete diploid set of chromosomes. It is the simplest form of polysomy and suitable for analyzing meiotic pairing and recombination patterns in situations where chromosomes compete for pairing. When no suitable meiotic chromosome markers are available, four metaphase I configurations can be distinguished. Their relative frequencies are indicative of the pairing and recombination patterns. In short arm (1RS) telotrisomics of chromosome 1R of rye (Secale cereale) we observed great differences in pairing and recombination patterns among spikes from different tillers and clones of the same plants. Anthers within spikes were only very rarely different. We analyzed a large number of genotypes, including inbreds as well as hybrids. The effects of genetic and environmental conditions on heterogeneity, if any, were limited. Considering that the reproductive tissue of a spike is derived from one primordial cell, it seems that at the start of sexual differentiation there was variation among cells in chromosomal control, which at meiosis determines pairing and crossing-over competence. We suggest that it is an epigenetic system that rigidly maintains this pattern through generative differentiation. In competitive situations the combination most competent for pairing will pair preferentially, forming specific meiotic configurations with different frequencies for different spikes of the same plant. This would explain the heterogeneity between spikes and the homogeneity within spikes. The epigenetic system could involve chromatin conformation or DNA methylation. There were no signs of heterochromatinization. PMID- 18430926 TI - Schizosaccharomyces pombe Hsp90/Git10 is required for glucose/cAMP signaling. AB - The fission yeast Schizosaccharomyces pombe senses environmental glucose through a cAMP-signaling pathway. Elevated cAMP levels activate protein kinase A (PKA) to inhibit transcription of genes involved in sexual development and gluconeogenesis, including the fbp1(+) gene, which encodes fructose-1,6 bisphosphatase. Glucose-mediated activation of PKA requires the function of nine glucose-insensitive transcription (git) genes, encoding adenylate cyclase, the PKA catalytic subunit, and seven "upstream" proteins required for glucose triggered adenylate cyclase activation. We describe the cloning and characterization of the git10(+) gene, which is identical to swo1(+) and encodes the S. pombe Hsp90 chaperone protein. Glucose repression of fbp1(+) transcription is impaired by both git10(-) and swo1(-) mutant alleles of the hsp90(+) gene, as well as by chemical inhibition of Hsp90 activity and temperature stress to wild type cells. Unlike the swo1(-) mutant alleles, the git10-201 allele supports cell growth at 37 degrees , while severely reducing glucose repression of an fbp1-lacZ reporter, suggesting a separation-of-function defect. Sequence analyses of three swo1(-) alleles and the one git10(-) allele indicate that swo1(-) mutations alter core functional domains of Hsp90, while the git10(-) mutation affects the Hsp90 central domain involved in client protein binding. These results suggest that Hsp90 plays a specific role in the S. pombe glucose/cAMP pathway. PMID- 18430927 TI - Distinct functions of MLH3 at recombination hot spots in the mouse. AB - The four mammalian MutL homologs (MLH1, MLH3, PMS1, and PMS2) participate in a variety of events, including postreplicative DNA repair, prevention of homeologous recombination, and crossover formation during meiosis. In this latter role, MLH1-MLH3 heterodimers predominate and are essential for prophase I progression. Previous studies demonstrated that mice lacking Mlh1 exhibit a 90% reduction in crossing over at the Psmb9 hot spot while noncrossovers, which do not result in exchange of flanking markers but arise from the same double-strand break event, are unaffected. Using a PCR-based strategy that allows for detailed analysis of crossovers and noncrossovers, we show here that Mlh3(-/-) exhibit a 85-94% reduction in the number of crossovers at the Psmb9 hot spot. Most of the remaining crossovers in Mlh3(-/-) meiocytes represent simple exchanges similar to those seen in wild-type mice, with a small fraction (6%) representing complex events that can extend far from the initiation zone. Interestingly, we detect an increase of noncrossovers in Mlh3(-/-) spermatocytes. These results suggest that MLH3 functions predominantly with MLH1 to promote crossovers, while noncrossover events do not require these activities. Furthermore, these results indicate that approximately 10% of crossovers in the mouse are independent of MLH3, suggesting the existence of alternative crossover pathways in mammals. PMID- 18430929 TI - Genetic analysis of the Caenorhabditis elegans GLH family of P-granule proteins. AB - The Vasa DEAD-box helicases are widespread markers of germ cells across species, and in some organisms have been shown to be essential for germ-cell formation and development. In contrast to the single Vasa gene in most systems analyzed, Caenorhabditis elegans has four Vasa family members, the germline helicases GLH 1, GLH-2, GLH-3, and GLH-4. Our analysis of deletion alleles of each glh gene demonstrates that GLH-1 is the key member of the family: loss of GLH-1 function causes sterility that is mainly maternal effect, is manifested predominantly at elevated temperature, and is due to reduced germ-cell proliferation and impaired formation of both sperm and oocytes. The other GLHs are not essential. However, GLH-4 serves redundant roles with GLH-1: loss of both genes' function causes glh 1-like sterility at all temperatures. Molecular epistasis analysis demonstrates that GLH-1 and GLH-4 are required for proper association of the PGL family of proteins with P granules, suggesting a pathway of P-granule assembly in which the GLHs are upstream of the PGL proteins and the mRNA cap-binding protein IFE-1. While loss of some P-granule components causes worms to be defective in RNA interference, loss of GLH-1 and GLH-4 does not compromise RNAi. Thus, RNAi likely does not require intact P granules but instead relies on particular P-granule factors. We discuss the evolution of the Vasa/GLH genes and current views of their functions and the assembly and roles of germ granules among species. PMID- 18430928 TI - Basolateral junctions utilize warts signaling to control epithelial-mesenchymal transition and proliferation crucial for migration and invasion of Drosophila ovarian epithelial cells. AB - Fasciclin2 (Fas2) and Discslarge (Dlg) localize to the basolateral junction (BLJ) of Drosophila follicle epithelial cells and inhibit their proliferation and invasion. To identify a BLJ signaling pathway we completed a genomewide screen for mutants that enhance dlg tumorigenesis. We identified two genes that encode known BLJ scaffolding proteins, lethal giant larvae (lgl) and scribble (scrib), and several not previously associated with BLJ function, including warts (wts) and roughened eye (roe), which encode a serine-threonine kinase and a transcription factor, respectively. Like scrib, wts and roe also enhance Fas2 and lgl tumorigenesis. Further, scrib, wts, and roe block border cell migration, and cause noninvasive tumors that resemble dlg partial loss of function, suggesting that the BLJ utilizes Wts signaling to repress EMT and proliferation, but not motility. Apicolateral junction proteins Fat (Ft), Expanded (Ex), and Merlin (Mer) either are not involved in these processes, or have highly spatio temporally restricted roles, diminishing their significance as upstream inputs to Wts in follicle cells. This is further indicated in that Wts targets, CyclinE and DIAP1, are elevated in Fas2, dlg, lgl, wts, and roe cells, but not Fat, ex, or mer cells. Thus, the BLJ appears to regulate epithelial polarity and dynamics not only as a localized scaffold, but also by communicating signals to the nucleus. Wts may be regulated by distinct junction inputs depending on developmental context. PMID- 18430930 TI - Raw mediates antagonism of AP-1 activity in Drosophila. AB - High baselines of transcription factor activities represent fundamental obstacles to regulated signaling. Here we show that in Drosophila, quenching of basal activator protein 1 (AP-1) transcription factor activity serves as a prerequisite to its tight spatial and temporal control by the JNK (Jun N-terminal kinase) signaling cascade. Our studies indicate that the novel raw gene product is required to limit AP-1 activity to leading edge epidermal cells during embryonic dorsal closure. In addition, we provide the first evidence that the epidermis has a Basket JNK-independent capacity to activate AP-1 targets and that raw function is required broadly throughout the epidermis to antagonize this activity. Finally, our mechanistic studies of the three dorsal-open group genes [raw, ribbon (rib), and puckered (puc)] indicate that these gene products provide at least two tiers of JNK/AP-1 regulation. In addition to Puckered phosphatase function in leading edge epidermal cells as a negative-feedback regulator of JNK signaling, the three dorsal-open group gene products (Raw, Ribbon, and Puckered) are required more broadly in the dorsolateral epidermis to quench a basal, signaling-independent activity of the AP-1 transcription factor. PMID- 18430931 TI - Regulation of glia number in Drosophila by Rap/Fzr, an activator of the anaphase promoting complex, and Loco, an RGS protein. AB - Glia mediate a vast array of cellular processes and are critical for nervous system development and function. Despite their immense importance in neurobiology, glia remain understudied and the molecular mechanisms that direct their differentiation are poorly understood. Rap/Fzr is the Drosophila homolog of the mammalian Cdh1, a regulatory subunit of the anaphase-promoting complex/cyclosome (APC/C). APC/C is an E3 ubiquitin ligase complex well characterized for its role in cell cycle progression. In this study, we have uncovered a novel cellular role for Rap/Fzr. Loss of rap/fzr function leads to a marked increase in the number of glia in the nervous system of third instar larvae. Conversely, ectopic expression of UAS-rap/fzr, driven by repo-GAL4, results in the drastic reduction of glia. Data from clonal analyses using the MARCM technique show that Rap/Fzr regulates the differentiation of surface glia in the developing larval nervous system. Our genetic and biochemical data further indicate that Rap/Fzr regulates glial differentiation through its interaction with Loco, a regulator of G-protein signaling (RGS) protein and a known effector of glia specification. We propose that Rap/Fzr targets Loco for ubiquitination, thereby regulating glial differentiation in the developing nervous system. PMID- 18430932 TI - Wispy, the Drosophila homolog of GLD-2, is required during oogenesis and egg activation. AB - Egg activation is the process that modifies mature, arrested oocytes so that embryo development can proceed. One key aspect of egg activation is the cytoplasmic polyadenylation of certain maternal mRNAs to permit or enhance their translation. wispy (wisp) maternal-effect mutations in Drosophila block development during the egg-to-embryo transition. We show here that the wisp gene encodes a member of the GLD-2 family of cytoplasmic poly(A) polymerases (PAPs). The WISP protein is required for poly(A) tail elongation of bicoid, Toll, and torso mRNAs upon egg activation. In Drosophila, WISP and Smaug (SMG) have previously been reported to be required to trigger the destabilization of maternal mRNAs during egg activation. SMG is the major regulator of this activity. We report here that SMG is still translated in activated eggs from wisp mutant mothers, indicating that WISP does not regulate mRNA stability by controlling the translation of smg mRNA. We have also analyzed in detail the very early developmental arrest associated with wisp mutations. Pronuclear migration does not occur in activated eggs laid by wisp mutant females. Finally, we find that WISP function is also needed during oogenesis to regulate the poly(A) tail length of dmos during oocyte maturation and to maintain a high level of active (phospho-) mitogen-activated protein kinases (MAPKs). PMID- 18430933 TI - Molecular characterization of Lal2, an SRK-like gene linked to the S-locus in the wild mustard Leavenworthia alabamica. AB - Single-locus sporophytic self-incompatibility inhibits inbreeding in many members of the mustard family (Brassicaceae). To investigate the genetics of self incompatibility in the wild mustard Leavenworthia alabamica, diallel crosses were conducted between full siblings. Patterns of incompatibility were consistent with the action of single-locus sporophytic self-incompatibility. DNA sequences related to S-locus receptor kinase (SRK), the gene involved in self-pollen recognition in mustards, were cloned and sequenced. A single sequence with high identity to SRK and several other groups of sequences (Lal1, Lal2, Lal3, Lal8, and Lal14) were isolated from L. alabamica. We propose that either Lal2 sequences are divergent alleles of SRK or Lal2 is in tight linkage with SRK because (1) Lal2 alleles cosegregate with S-alleles inferred from dialleles in all 97 cases tested in five families; (2) Lal2 sequences are highly diverse at both synonymous and nonsynonymous sites and exhibit patterns of selective constraint similar to those observed at SRK in Brassica and Arabidopsis; and (3) transcripts of one Lal2 allele were detected in leaves and the styles of open flowers, but were most abundant in the stigmas of maturing buds. We discuss the utility of the S-linked polymorphism at Lal2 for studying the evolutionary forces acting on self incompatibility in Leavenworthia. PMID- 18430934 TI - Fraction of informative recombinations: a heuristic approach to analyze recombination rates. AB - In this article we present a new heuristic approach (informative recombinations, InfRec) to analyze recombination density at the sequence level. InfRec is intuitive and easy and combines previously developed methods that (i) resolve genotypes into haplotypes, (ii) estimate the minimum number of recombinations, and (iii) evaluate the fraction of informative recombinations. We tested this approach in its sliding-window version on 117 genes from the SeattleSNPs program, resequenced in 24 African-Americans (AAs) and 23 European-Americans (EAs). We obtained population recombination rate estimates (rho(obs)) of 0.85 and 0.37 kb( 1) in AAs and EAs, respectively. Coalescence simulations indicated that these values account for both the recombinations and the gene conversions in the history of the sample. The intensity of rho(obs) varied considerably along the sequence, revealing the presence of recombination hotspots. Overall, we observed approximately 80% of recombinations in one-third and approximately 50% in only 10% of the sequence. InfRec performance, tested on published simulated and additional experimental data sets, was similar to that of other hotspot detection methods. Fast, intuitive, and visual, InfRec is not constrained by sample size limitations. It facilitates understanding data and provides a simple and flexible tool to analyze recombination intensity along the sequence. PMID- 18430935 TI - Patterns of molecular evolution in Caenorhabditis preclude ancient origins of selfing. AB - The evolution of self-fertilization can mediate pronounced changes in genomes as a by-product of a drastic reduction in effective population size and the concomitant accumulation of slightly deleterious mutations by genetic drift. In the nematode genus Caenorhabditis, a highly selfing lifestyle has evolved twice independently, thus permitting an opportunity to test for the effects of mode of reproduction on patterns of molecular evolution on a genomic scale. Here we contrast rates of nucleotide substitution and codon usage bias among thousands of orthologous groups of genes in six species of Caenorhabditis, including the classic model organism Caenorhabditis elegans. Despite evidence that weak selection on synonymous codon usage is pervasive in the history of all species in this genus, we find little difference among species in the patterns of codon usage bias and in replacement-site substitution. Applying a model of relaxed selection on codon usage to the C. elegans and C. briggsae lineages suggests that self-fertilization is unlikely to have evolved more than approximately 4 million years ago, which is less than a quarter of the time since they shared a common ancestor with outcrossing species. We conclude that the profound changes in mating behavior, physiology, and developmental mechanisms that accompanied the transition from an obligately outcrossing to a primarily selfing mode of reproduction evolved in the not-too-distant past. PMID- 18430936 TI - Interactions between stressful environment and gene deletions alleviate the expected average loss of fitness in yeast. AB - The conjecture that the deleterious effects of mutations are amplified by stress or interaction with one another remains unsatisfactorily tested. It is now possible to reapproach this problem systematically by using genomic collections of mutants and applying stress-inducing conditions with a well-recognized impact on metabolism. We measured the maximum growth rate of single- and double-gene deletion strains of yeast in several stress-inducing treatments, including poor nutrients, elevated temperature, high salinity, and the addition of caffeine. The negative impact of deletions on the maximum growth rate was relatively smaller in stressful than in favorable conditions. In both benign and harsh environments, double-deletion strains grew on average slightly faster than expected from a multiplicative model of interaction between single growth effects, indicating positive epistasis for the rate of growth. This translates to even higher positive epistasis for fitness defined as the number of progeny. We conclude that the negative impact of metabolic disturbances, regardless of whether they are of environmental or genetic origin, is absolutely and relatively highest when growth is fastest. The effect of further damages tends to be weaker. This results in an average alleviating effect of interactions between stressful environment and gene deletions and among gene deletions. PMID- 18430937 TI - The rate and spectrum of microsatellite mutation in Caenorhabditis elegans and Daphnia pulex. AB - The effective use of microsatellite loci as tools for microevolutionary analysis requires knowledge of the factors influencing the rate and pattern of mutation, much of which is derived from indirect inference from population samples. Interspecific variation in microsatellite stability also provides a glimpse into aspects of phylogenetic constancy of mutational processes. Using long-term series of mutation-accumulation lines, we have obtained direct estimates of the spectrum of microsatellite mutations in two model systems: the nematode Caenorhabditis elegans and the microcrustacean Daphnia pulex. Although the scaling of the mutation rate with the number of tandem repeats is highly consistent across distantly related species, including yeast and human, the per-cell-division mutation rate appears to be elevated in multicellular species. Contrary to the expectations under the stepwise mutation model, most microsatellite mutations in C. elegans and D. pulex involve changes of multiple repeat units, with expansions being much more common than contractions. PMID- 18430938 TI - Estimation of pairwise identity by descent from dense genetic marker data in a population sample of haplotypes. AB - I present a new approach for calculating probabilities of identity by descent for pairs of haplotypes. The approach is based on a joint hidden Markov model for haplotype frequencies and identity by descent (IBD). This model allows for linkage disequilibrium, and the method can be applied to very dense marker data. The method has high power for detecting IBD tracts of genetic length of 1 cM, with the use of sufficiently dense markers. This enables detection of pairwise IBD between haplotypes from individuals whose most recent common ancestor lived up to 50 generations ago. PMID- 18430939 TI - The role of regulatory genes during maize domestication: evidence from nucleotide polymorphism and gene expression. AB - We investigated DNA sequence variation in 72 candidate genes in maize landraces and the wild ancestor of maize, teosinte. The candidate genes were chosen because they exhibit very low sequence diversity among maize inbreds and have sequence homology to known regulatory genes. We observed signatures of selection in 17 candidate genes, indicating that they were potential targets of artificial selection during domestication. In addition, 21 candidate genes were identified as potential targets of natural selection in teosinte. A comparison of the proportion of selected genes between our regulatory genes and genes unfiltered for their potential function (but also with very low sequence diversity among maize inbreds) provided some weak evidence that regulatory genes are overrepresented among selected genes. We detected no significant association between the positions of genes identified as potential targets of selection during domestication and quantitative trait loci (QTL) responsible for maize domestication traits. However, a subset of these genes, those identified by sequence homology as kinase/phosphatase genes, significantly cluster with the domestication QTL. We also analyzed expression profiles of genes in distinct maize tissues and observed that domestication genes are expressed on average at a significantly higher level than neutral genes in reproductive organs, including kernels. PMID- 18430940 TI - Multiple rescue factors within a Wolbachia strain. AB - Wolbachia-induced cytoplasmic incompatibility (CI) is expressed when infected males are crossed with either uninfected females or females infected with Wolbachia of different CI specificity. In diploid insects, CI results in embryonic mortality, apparently due to the the loss of the paternal set of chromosomes, usually during the first mitotic division. The molecular basis of CI has not been determined yet; however, several lines of evidence suggest that Wolbachia exhibits two distinct sex-dependent functions: in males, Wolbachia somehow "imprints" the paternal chromosomes during spermatogenesis (mod function), whereas in females, the presence of the same Wolbachia strain(s) is able to restore embryonic viability (resc function). On the basis of the ability of Wolbachia to induce the modification and/or rescue functions in a given host, each bacterial strain can be classified as belonging in one of the four following categories: mod(+) resc(+), mod(-) resc(+), mod(-) resc(-), and mod(+) resc(-). A so-called "suicide" mod(+) resc(-) strain has not been found in nature yet. Here, a combination of embryonic cytoplasmic injections and introgression experiments was used to transfer nine evolutionary, distantly related Wolbachia strains (wYak, wTei, wSan, wRi, wMel, wHa, wAu, wNo, and wMa) into the same host background, that of Drosophila simulans (STCP strain), a highly permissive host for CI expression. We initially characterized the modification and rescue properties of the Wolbachia strains wYak, wTei, and wSan, naturally present in the yakuba complex, upon their transfer into D. simulans. Confocal microscopy and multilocus sequencing typing (MLST) analysis were also employed for the evaluation of the CI properties. We also tested the compatibility relationships of wYak, wTei, and wSan with all other Wolbachia infections. So far, the cytoplasmic incompatibility properties of different Wolbachia variants are explained assuming a single pair of modification and rescue factors specific to each variant. This study shows that a given Wolbachia variant can possess multiple rescue determinants corresponding to different CI systems. In addition, our results: (a) suggest that wTei appears to behave in D. simulans as a suicide mod(+) resc(-) strain, (b) unravel unique CI properties, and (c) provide a framework to understand the diversity and the evolution of new CI-compatibility types. PMID- 18430942 TI - The effects of recombination rate on the distribution and abundance of transposable elements. AB - Transposable elements (TEs) often accumulate in regions of the genome with suppressed recombination. But it is unclear whether this pattern reflects a reduction in the efficacy of selection against deleterious insertions or a relaxation of ectopic recombination. Discriminating between these two hypotheses has been difficult, because no formal model has investigated the effects of recombination under the deleterious insertion model. Here we take a simulation based approach to analyze this scenario and determine the conditions under which element accumulation is expected in low recombination regions. We show that TEs become fixed as a result of Hill-Robertson effects in the form of Muller's ratchet, but only in regions of extremely low recombination when excision is effectively absent and synergism between elements is weak. These results have important implications for differentiating between the leading models of how selection acts on TEs and should help to interpret emerging population genetic and genomic data. PMID- 18430941 TI - Rapid evolution of yeast centromeres in the absence of drive. AB - To find the most rapidly evolving regions in the yeast genome we compared most of chromosome III from three closely related lineages of the wild yeast Saccharomyces paradoxus. Unexpectedly, the centromere appears to be the fastest evolving part of the chromosome, evolving even faster than DNA sequences unlikely to be under selective constraint (i.e., synonymous sites after correcting for codon usage bias and remnant transposable elements). Centromeres on other chromosomes also show an elevated rate of nucleotide substitution. Rapid centromere evolution has also been reported for some plants and animals and has been attributed to selection for inclusion in the egg or the ovule at female meiosis. But Saccharomyces yeasts have symmetrical meioses with all four products surviving, thus providing no opportunity for meiotic drive. In addition, yeast centromeres show the high levels of polymorphism expected under a neutral model of molecular evolution. We suggest that yeast centromeres suffer an elevated rate of mutation relative to other chromosomal regions and they change through a process of "centromere drift," not drive. PMID- 18430943 TI - Adaptive plasmid evolution results in host-range expansion of a broad-host-range plasmid. AB - Little is known about the range of hosts in which broad-host-range (BHR) plasmids can persist in the absence of selection for plasmid-encoded traits, and whether this "long-term host range" can evolve over time. Previously, the BHR multidrug resistance plasmid pB10 was shown to be highly unstable in Stenotrophomonas maltophilia P21 and Pseudomonas putida H2. To investigate whether this plasmid can adapt to such unfavorable hosts, we performed evolution experiments wherein pB10 was maintained in strain P21, strain H2, and alternatingly in P21 and H2. Plasmids that evolved in P21 and in both hosts showed increased stability and decreased cost in ancestral host P21. However, the latter group showed higher variability in stability patterns, suggesting that regular switching between distinct hosts hampered adaptive plasmid evolution. The plasmids evolved in P21 were also equally or more stable in other hosts compared to pB10, which suggested true host-range expansion. The complete genome sequences of four evolved plasmids with improved stability showed only one or two genetic changes. The stability of plasmids evolved in H2 improved only in their coevolved hosts, not in the ancestral host. Thus a BHR plasmid can adapt to an unfavorable host and thereby expand its long-term host range. PMID- 18430944 TI - Genetic variation affecting host-parasite interactions: different genes affect different aspects of sigma virus replication and transmission in Drosophila melanogaster. AB - In natural populations, genetic variation affects resistance to disease. Knowing how much variation exists, and understanding the genetic architecture of this variation, is important for medicine, for agriculture, and for understanding evolutionary processes. To investigate the extent and nature of genetic variation affecting resistance to pathogens, we are studying a tractable model system: Drosophila melanogaster and its natural pathogen the vertically transmitted sigma virus. We show that considerable genetic variation affects transmission of the virus from parent to offspring. However, maternal and paternal transmission of the virus is affected by different genes. Maternal transmission is a simple Mendelian trait: most of the genetic variation is explained by a polymorphism in ref(2)P, a gene already well known to affect resistance to sigma. In contrast, there is considerable genetic variation in paternal transmission that cannot be explained by ref(2)P and is caused by other loci on chromosome 2. Furthermore, we found no genetic correlation between paternal transmission of the virus and resistance to infection by the sigma virus following injection. This suggests that different loci affect viral replication and paternal transmission. PMID- 18430945 TI - Fine mapping of quantitative trait loci affecting female fertility in dairy cattle on BTA03 using a dense single-nucleotide polymorphism map. AB - Fertility quantitative trait loci (QTL) are of high interest in dairy cattle since insemination failure has dramatically increased in some breeds such as Holstein. High-throughput SNP analysis and SNP microarrays give the opportunity to genotype many animals for hundreds SNPs per chromosome. In this study, due to these techniques a dense SNP marker map was used to fine map a QTL underlying nonreturn rate measured 90 days after artificial insemination previously detected with a low-density microsatellite marker map. A granddaughter design with 17 Holstein half-sib families (926 offspring) was genotyped for a set of 437 SNPs mapping to BTA3. Linkage analysis was performed by both regression and variance components analysis. An additional analysis combining both linkage analysis and linkage-disequilibrium information was applied. This method first estimated identity-by-descent probabilities among base haplotypes. These probabilities were then used to group the base haplotypes in different clusters. A QTL explaining 14% of the genetic variance was found with high significance (P < 0.001) at position 19 cM with the linkage analysis and four sires were estimated to be heterozygous (P < 0.05). Addition of linkage-disequilibrium information refined the QTL position to a set of narrow peaks. The use of the haplotypes of heterozygous sires offered the possibility to give confidence in some peaks while others could be discarded. Two peaks with high likelihood-ratio test values in the region of which heterozygous sires shared a common haplotype appeared particularly interesting. Despite the fact that the analysis did not fine map the QTL in a unique narrow region, the method proved to be able to handle efficiently and automatically a large amount of information and to refine the QTL position to a small set of narrow intervals. In addition, the QTL identified was confirmed to have a large effect (explaining 13.8% of the genetic variance) on dairy cow fertility as estimated by nonreturn rate at 90 days. PMID- 18430946 TI - Semidominant mutations in reduced epidermal fluorescence 4 reduce phenylpropanoid content in Arabidopsis. AB - Plants synthesize an array of natural products that play diverse roles in growth, development, and defense. The plant-specific phenylpropanoid metabolic pathway produces as some of its major products flavonoids, monolignols, and hydroxycinnamic- acid conjugates. The reduced epidermal fluorescence 4 (ref4) mutant is partially dwarfed and accumulates reduced quantities of all phenylpropanoid-pathway end products. Further, plants heterozygous for ref4 exhibit intermediate growth and phenylpropanoid-related phenotypes, suggesting that these mutations are semidominant. The REF4 locus (At2g48110) was cloned by a combined map- and sequencing-based approach and was found to encode a large integral membrane protein that is unique to plants. The mutations in all ref4 alleles cause substitutions in conserved amino acids that are located adjacent to predicted transmembrane regions. Expression of the ref4-3 allele in wild-type and null REF4 plants caused reductions in sinapoylmalate content, lignin content, and growth, demonstrating that the mutant alleles are truly semidominant. Further, a suppressor mutant was isolated that abolishes a WW protein-protein interaction domain that may be important for REF4 function. PMID- 18430947 TI - Quantitative trait loci mapping in five new large recombinant inbred line populations of Arabidopsis thaliana genotyped with consensus single-nucleotide polymorphism markers. AB - Quantitative approaches conducted in a single mapping population are limited by the extent of genetic variation distinguishing the parental genotypes. To overcome this limitation and allow a more complete dissection of the genetic architecture of complex traits, we built an integrated set of 15 new large Arabidopsis thaliana recombinant inbred line (RIL) populations optimized for quantitative trait loci (QTL) mapping, having Columbia as a common parent crossed to distant accessions. Here we present 5 of these populations that were validated by investigating three traits: flowering time, rosette size, and seed production as an estimate of fitness. The large number of RILs in each population (between 319 and 377 lines) and the high density of evenly spaced genetic markers scored ensure high power and precision in QTL mapping even under a minimal phenotyping framework. Moreover, the use of common markers across the different maps allows a direct comparison of the QTL detected within the different RIL sets. In addition, we show that following a selective phenotyping strategy by performing QTL analyses on genotypically chosen subsets of 164 RILs (core populations) does not impair the power of detection of QTL with phenotypic contributions >7%. PMID- 18430948 TI - Genetic expectations of quantitative trait loci main and interaction effects obtained with the triple testcross design and their relevance for the analysis of heterosis. AB - Interpretation of experimental results from quantitative trait loci (QTL) mapping studies on the predominant type of gene action can be severely affected by the choice of statistical model, experimental design, and provision of epistasis. In this study, we derive quantitative genetic expectations of (i) QTL effects obtained from one-dimensional genome scans with the triple testcross (TTC) design and (ii) pairwise interactions between marker loci using two-way analyses of variance (ANOVA) under the F(2)- and the F(infinity)-metric model. The theoretical results show that genetic expectations of QTL effects estimated with the TTC design are complex, comprising both main and epistatic effects, and that genetic expectations of two-way marker interactions are not straightforward extensions of effects estimated in one-dimensional scans. We also demonstrate that the TTC design can partially overcome the limitations of the design III in separating QTL main effects and their epistatic interactions in the analysis of heterosis and that dominance x additive epistatic interactions of individual QTL with the genetic background can be estimated with a one-dimensional genome scan. Furthermore, we present genetic expectations of variance components for the analysis of TTC progeny tested in a split-plot design, assuming digenic epistasis and arbitrary linkage. PMID- 18430949 TI - Pleiotropic patterns of quantitative trait loci for 70 murine skeletal traits. AB - Quantitative trait locus (QTL) studies of a skeletal trait or a few related skeletal components are becoming commonplace, but as yet there has been no investigation of pleiotropic patterns throughout the skeleton. We present a comprehensive survey of pleiotropic patterns affecting mouse skeletal morphology in an intercross of LG/J and SM/J inbred strains (N = 1040), using QTL analysis on 70 skeletal traits. We identify 798 single-trait QTL, coalescing to 105 loci that affect on average 7-8 traits each. The number of traits affected per locus ranges from only 1 trait to 30 traits. Individual traits average 11 QTL each, ranging from 4 to 20. Skeletal traits are affected by many, small-effect loci. Significant additive genotypic values average 0.23 standard deviation (SD) units. Fifty percent of loci show codominance with heterozygotes having intermediate phenotypic values. When dominance does occur, the LG/J allele tends to be dominant to the SM/J allele (30% vs. 8%). Over- and underdominance are relatively rare (12%). Approximately one-fifth of QTL are sex specific, including many for pelvic traits. Evaluating the pleiotropic relationships of skeletal traits is important in understanding the role of genetic variation in the growth and development of the skeleton. PMID- 18430950 TI - Reproducing kernel hilbert spaces regression methods for genomic assisted prediction of quantitative traits. AB - Reproducing kernel Hilbert spaces regression procedures for prediction of total genetic value for quantitative traits, which make use of phenotypic and genomic data simultaneously, are discussed from a theoretical perspective. It is argued that a nonparametric treatment may be needed for capturing the multiple and complex interactions potentially arising in whole-genome models, i.e., those based on thousands of single-nucleotide polymorphism (SNP) markers. After a review of reproducing kernel Hilbert spaces regression, it is shown that the statistical specification admits a standard mixed-effects linear model representation, with smoothing parameters treated as variance components. Models for capturing different forms of interaction, e.g., chromosome-specific, are presented. Implementations can be carried out using software for likelihood-based or Bayesian inference. PMID- 18430951 TI - Nonparametric methods for incorporating genomic information into genetic evaluations: an application to mortality in broilers. AB - Four approaches using single-nucleotide polymorphism (SNP) information (F(infinity)-metric model, kernel regression, reproducing kernel Hilbert spaces (RKHS) regression, and a Bayesian regression) were compared with a standard procedure of genetic evaluation (E-BLUP) of sires using mortality rates in broilers as a response variable, working in a Bayesian framework. Late mortality (14-42 days of age) records on 12,167 progeny of 200 sires were precorrected for fixed and random (nongenetic) effects used in the model for genetic evaluation and for the mate effect. The average of the corrected records was computed for each sire. Twenty-four SNPs seemingly associated with late mortality were included in three methods used for genomic assisted evaluations. One thousand SNPs were included in the Bayesian regression, to account for markers along the whole genome. The posterior mean of heritability of mortality was 0.02 in the E BLUP approach, suggesting that genetic evaluation could be improved if suitable molecular markers were available. Estimates of posterior means and standard deviations of the residual variance were 24.38 (3.88), 29.97 (3.22), 17.07 (3.02), and 20.74 (2.87) for E-BLUP, the linear model on SNPs, RKHS regression, and the Bayesian regression, respectively, suggesting that RKHS accounted for more variance in the data. The two nonparametric methods (kernel and RKHS regression) fitted the data better, having a lower residual sum of squares. Predictive ability, assessed by cross-validation, indicated advantages of the RKHS approach, where accuracy was increased from 25 to 150%, relative to other methods. PMID- 18430952 TI - An improved method for quantitative trait loci detection and identification of within-line segregation in F2 intercross designs. AB - We present a new flexible, simple, and powerful genome-scan method (flexible intercross analysis, FIA) for detecting quantitative trait loci (QTL) in experimental line crosses. The method is based on a pure random-effects model that simultaneously models between- and within-line QTL variation for single as well as epistatic QTL. It utilizes the score statistic and thereby facilitates computationally efficient significance testing based on empirical significance thresholds obtained by means of permutations. The properties of the method are explored using simulations and analyses of experimental data. The simulations showed that the power of FIA was as good as, or better than, Haley-Knott regression and that FIA was rather insensitive to the level of allelic fixation in the founders, especially for pedigrees with few founders. A chromosome scan was conducted for a meat quality trait in an F(2) intercross in pigs where a mutation in the halothane (Ryanodine receptor, RYR1) gene with a large effect on meat quality was known to segregate in one founder line. FIA obtained significant support for the halothane-associated QTL and identified the base generation allele with the mutated allele. A genome scan was also performed in a previously analyzed chicken F(2) intercross. In the chicken intercross analysis, four previously detected QTL were confirmed at a 5% genomewide significance level, and FIA gave strong evidence (P < 0.01) for two of these QTL to be segregating within the founder lines. FIA was also extended to account for epistasis and using simulations we show that the method provides good estimates of epistatic QTL variance even for segregating QTL. Extensions of FIA and its applications on other intercross populations including backcrosses, advanced intercross lines, and heterogeneous stocks are also discussed. PMID- 18430953 TI - High diversity of genes for nonhost resistance of barley to heterologous rust fungi. AB - Inheritance studies on the nonhost resistance of plants would normally require interspecific crosses that suffer from sterility and abnormal segregation. Therefore, we developed the barley-Puccinia rust model system to study, using forward genetics, the specificity, number, and diversity of genes involved in nonhost resistance. We developed two mapping populations by crossing the line SusPtrit, with exceptional susceptibility to heterologous rust species, with the immune barley cultivars Vada and Cebada Capa. These two mapping populations along with the Oregon Wolfe Barley population, which showed unexpected segregation for resistance to heterologous rusts, were phenotyped with four heterologous rust fungal species. Positions of QTL conferring nonhost resistance in the three mapping populations were compared using an integrated consensus map. The results confirmed that nonhost resistance in barley to heterologous rust species is controlled by QTL with different and overlapping specificities and by an occasional contribution of an R-gene for hypersensitivity. In each population, different sets of loci were implicated in resistance. Few genes were common between the populations, suggesting a high diversity of genes conferring nonhost resistance to heterologous pathogens. These loci were significantly associated with QTL for partial resistance to the pathogen Puccinia hordei and with defense related genes. PMID- 18430954 TI - Quantitative genetic analysis of sleep in Drosophila melanogaster. AB - Although intensively studied, the biological purpose of sleep is not known. To identify candidate genes affecting sleep, we assayed 136 isogenic P-element insertion lines of Drosophila melanogaster. Since sleep has been negatively correlated with energy reserves across taxa, we measured energy stores (whole body protein, glycogen, and triglycerides) in these lines as well. Twenty-one insertions with known effects on physiology, development, and behavior affect 24 hr sleep time. Thirty-two candidate insertions significantly impact energy stores. Mutational genetic correlations among sleep parameters revealed that the genetic basis of the transition between sleep and waking states in males and females may be different. Furthermore, sleep bout number can be decoupled from waking activity in males, but not in females. Significant genetic correlations are present between sleep phenotypes and glycogen stores in males, while sleep phenotypes are correlated with triglycerides in females. Differences observed in male and female sleep behavior in flies may therefore be related to sex-specific differences in metabolic needs. Sleep thus emerges as a complex trait that exhibits extensive pleiotropy and sex specificity. The large mutational target that we observed implicates genes functioning in a variety of biological processes, suggesting that sleep may serve a number of different functions rather than a single purpose. PMID- 18430955 TI - The protease activity of yeast separase (esp1) is required for anaphase spindle elongation independently of its role in cleavage of cohesin. AB - Separase is a caspase-family protease required for the metaphase-anaphase transition in eukaryotes. In budding yeast, the separase ortholog, Esp1, has been shown to cleave a subunit of cohesin, Mcd1 (Scc1), thereby releasing sister chromatids from cohesion and allowing anaphase. However, whether Esp1 has other substrates required for anaphase has been controversial. Whereas it has been reported that cleavage of Mcd1 is sufficient to trigger anaphase in the absence of Esp1 activation, another study using a temperature-sensitive esp1 mutant concluded that depletion of Mcd1 was not sufficient for anaphase in the absence of Esp1 function. Here we revisit the issue and demonstrate that neither depletion of Mcd1 nor ectopic cleavage of Mcd1 by Tev1 protease is sufficient to support anaphase in an esp1 temperature-sensitive mutant. Furthermore, we demonstrate that the catalytic activity of the Esp1 protease is required for this Mcd1-independent anaphase function. These data suggest that another protein, possibly a spindle-associated protein, is cleaved by Esp1 to allow anaphase. Such a function is consistent with the previous observation that Esp1 localizes to the mitotic spindle during anaphase. PMID- 18430956 TI - Functional conservation of the yeast and Arabidopsis RAD54-like genes. AB - The Saccharomyces cerevisiae RAD54 gene has critical roles in DNA double-strand break repair, homologous recombination, and gene targeting. Previous results show that the yeast gene enhances gene targeting when expressed in Arabidopsis thaliana. In this work we address the trans-species compatibility of Rad54 functions. We show that overexpression of yeast RAD54 in Arabidopsis enhances DNA damage resistance severalfold. Thus, the yeast gene is active in the Arabidopsis homologous-recombination repair system. Moreover, we have identified an A. thaliana ortholog of yeast RAD54, named AtRAD54. This gene, with close sequence similarity to RAD54, complements methylmethane sulfonate (MMS) sensitivity but not UV sensitivity or gene targeting defects of rad54Delta mutant yeast cells. Overexpression of AtRAD54 in Arabidopsis leads to enhanced resistance to DNA damage. This gene's assignment as a RAD54 ortholog is further supported by the interaction of AtRad54 with AtRad51 and the interactions between alien proteins (i.e., yeast Rad54 with AtRAD51 and yeast Rad51 with AtRad54) in a yeast two hybrid experiment. These interactions hint at the molecular nature of this interkingdom complementation, although the stronger effect of the yeast Rad54 in plants than AtRad54 in yeast might be explained by an ability of the Rad54 protein to act alone, independently of its interaction with Rad51. PMID- 18430957 TI - The Rad52 homologs Rad22 and Rti1 of Schizosaccharomyces pombe are not essential for meiotic interhomolog recombination, but are required for meiotic intrachromosomal recombination and mating-type-related DNA repair. AB - Proteins of the RAD52 epistasis group play an essential role in repair of some types of DNA damage and genetic recombination. In Schizosaccharomyces pombe, Rad22 (a Rad52 ortholog) has been shown to be as necessary for repair and recombination events during vegetative growth as its Saccharomyces cerevisiae counterpart. This finding contrasts with previous reports where, due to suppressor mutations in the fbh1 gene, rad22 mutants did not display a severe defect. We have analyzed the roles of Rad22 and Rti1, another Rad52 homolog, during meiotic recombination and meiosis in general. Both proteins play an important role in spore viability. During meiotic prophase I, they partially colocalize and partially localize to Rad51 foci and linear elements. Genetic analysis showed that meiotic interchromosomal crossover and conversion events were unexpectedly not much affected by deletion of either or both genes. A strong decrease of intrachromosomal recombination assayed by a gene duplication construct was observed. Therefore, we propose that the most important function of Rad22 and Rti1 in S. pombe meiosis is repair of double-strand breaks with involvement of the sister chromatids. In addition, a novel mating-type-related repair function of Rad22 specific to meiosis and spore germination is described. PMID- 18430958 TI - Visual mutations reveal opposing effects of illumination on arousal in Drosophila. AB - The effect of illumination on alertness can be assessed by comparing the efficacy of an anesthetic under light vs. dark conditions. Results from such tests on wild type flies and visual mutants demonstrate that, surprisingly, light has both positive and negative influences on arousal. These dual effects may explain aspects of the fly's daily activity and have potential clinical implications. PMID- 18430959 TI - Exploring population genetic models with recombination using efficient forward time simulations. AB - We present an exact forward-in-time algorithm that can efficiently simulate the evolution of a finite population under the Wright-Fisher model. We used simulations based on this algorithm to verify the accuracy of the ancestral recombination graph approximation by comparing it to the exact Wright-Fisher scenario. We find that the recombination graph is generally a very good approximation for models with complete outcrossing, whereas, for models with self fertilization, the approximation becomes slightly inexact for some combinations of selfing and recombination parameters. PMID- 18430960 TI - Nearly neutrality and the evolution of codon usage bias in eukaryotic genomes. AB - Here I show that the mean codon usage bias of a genome, and of the lowly expressed genes in a genome, is largely similar across eukaryotes ranging from unicellular protists to vertebrates. Conversely, this bias in housekeeping genes and in highly expressed genes has a remarkable inverse relationship with species generation time that varies by more than four orders of magnitude. The relevance of these results to the nearly neutral theory of molecular evolution is discussed. PMID- 18430961 TI - Key impact of Vgt1 on flowering time adaptation in maize: evidence from association mapping and ecogeographical information. AB - An association study conducted on 375 maize inbred lines indicates a strong relationship between Vgt1 polymorphisms and flowering time, extending former quantitative trait loci (QTL) mapping results. Analysis of allele frequencies in a landrace collection supports a key role of Vgt1 in maize altilatitudinal adaptation. PMID- 18430962 TI - Frontiers: skeletal muscle sodium pump regulation: a translocation paradigm. AB - The skeletal muscle sodium pump plays a major role in the removal of K(+) ions from the circulation postprandial, or after a physical activity bout, thereby preventing the development of hyperkalemia and fatigue. Insulin and muscle contractions stimulate Na(+)-K(+)-ATPase activity in skeletal muscle, at least partially via translocation of sodium pump units to the plasma membrane from intracellular stores. The molecular mechanism of this phenomenon is poorly understood. Due to the contradictory reports in the literature, the very existence of the translocation of Na(+)-K(+)-ATPase to the skeletal muscle cell surface is questionable. This review summarizes more than 30 years work on the skeletal muscle sodium pump translocation paradigm. Furthermore, the methodological caveats of major approaches to study the sodium pump translocation in skeletal muscle are discussed. An understanding of the molecular regulation of Na(+)-K(+)-ATPase in skeletal muscle will have important clinical implications for the understanding of the development of complications associated with the metabolic syndrome, such as cardiovascular diseases or increased muscle fatigue in diabetic patients. PMID- 18430964 TI - Adipose tissue inflammation and liver fat in patients with highly active antiretroviral therapy-associated lipodystrophy. AB - In this cross-sectional study, we sought to determine whether gene expression of macrophage markers and inflammatory chemokines in lipoatrophic subcutaneous abdominal adipose tissue and liver fat content are increased and interrelated in human immunodeficiency virus (HIV)-1-positive, highly active antiretroviral therapy (HAART)-treated patients with lipodystrophy (HAART+LD+; n = 27) compared with those without (HAART+LD-; n = 13). The study groups were comparable with respect to age, gender, and body mass index. The HAART+LD+ group had twofold more intra-abdominal (P = 0.01) and 1.5-fold less subcutaneous (P = 0.091) fat than the HAART+LD- group. As we have reported previously, liver fat was 10-fold higher in the HAART+LD+ compared with the HAART+LD- group (P = 0.00003). Inflammatory gene expression was increased in HAART-lipodystrophy: CD68 4.5-fold (P = 0.000013), tumor necrosis factor (TNF)-alpha 2-fold (P = 0.0094), chemokine (C-C motif) ligand (CCL) 2 2.5-fold (P = 0.0024), CCL3 7-fold (P = 0.0000017), integrin alphaM (ITGAM) 3-fold (P = 0.00067), epidermal growth factor-like module containing, mucin-like, hormone receptor-like (EMR)1 2.5-fold (P = 0.0038), and a disintegrin and metalloproteinase domain (ADAM)8 3.5-fold (P = 0.00057) higher in the HAART+LD+ compared with the HAART+LD- group. mRNA concentration of CD68 (r = 0.37, P = 0.019), ITGAM (r = 0.35, P = 0.025), CCL2 (r = 0.39, P = 0.012), and CCL3 (r = 0.54, P = 0.0003) correlated with liver fat content. In conclusion, gene expression of markers of macrophage infiltration and adipose tissue inflammation is increased in lipoatrophic subcutaneous abdominal adipose tissue of patients with HAART-associated lipodystrophy compared with those without. CD68, ITGAM, CCL2, and CCL3 expression is significantly associated with accumulation of liver fat. PMID- 18430963 TI - Differential modulation of L-type calcium channel subunits by oleate. AB - Nonesterified fatty acids such as oleate and palmitate acutely potentiate insulin secretion from pancreatic islets in a glucose-dependent manner. In addition, recent studies show that fatty acids elevate intracellular free Ca(2+) and increase voltage-gated Ca(2+) current in mouse beta-cells, although the mechanisms involved are poorly understood. Here we utilized a heterologous system to express subunit-defined voltage-dependent L-type Ca(2+) channels (LTCC) and demonstrate that beta-cell calcium may increase in part from an interaction between fatty acid and specific calcium channel subunits. Distinct functional LTCC were assembled in both COS-7 and HEK-293 cells by expressing either one of the EYFP-tagged L-type alpha(1)-subunits (beta-cell Cav1.3 or lung Cav1.2) and ERFP-tagged islet beta-subunits (ibeta(2a) or ibeta(3)). In COS-7 cells, elevations in intracellular Ca(2+) mediated by LTCC were enhanced by an oleate BSA complex. To extend these findings, Ca(2+) current was measured in LTCC expressing HEK-293 cells that revealed an increase in peak Ca(2+) current within 2 min after addition of the oleate complex, with maximal potentiation occurring at voltages <0 mV. Both Cav1.3 and Cav1.2 were modulated by oleate, and the presence of different auxiliary beta-subunits resulted in differential augmentation. The potentiating effect of oleate on Cav1.2 was abolished by the pretreatment of cells with triacsin C, suggesting that long-chain CoA synthesis is necessary for Ca(2+) channel modulation. These results show for the first time that two L-type Ca(2+) channels expressed in beta-cells (Cav1.3 and Cav1.2) appear to be targeted by nonesterified fatty acids. This effect may account in part for the acute potentiation of glucose-dependent insulin secretion by fatty acids. PMID- 18430965 TI - Effects of a supraphysiological dose of testosterone on physical function, muscle performance, mood, and fatigue in men with HIV-associated weight loss. AB - Testosterone increases fat-free mass (FFM) in men infected with human immunodeficiency virus (HIV), but its effects on muscle performance, physical function, mood, and quality of life are poorly understood. Sixty-one HIV-infected men with weight loss were randomized to receive weekly intramuscular injections of 300 mg of testosterone enanthate or placebo for 16 wk. The primary outcome of interest was physical function (walking speed, stair-climbing power, and load carrying ability). Secondary outcome measures included body weight and composition, muscle performance, sexual function, mood, and quality of life. Serum nadir free and total testosterone levels increased (+188.0 +/- 29.6 and +720 +/- 86 ng/dl) in the testosterone, but not placebo, group. Testosterone administration was associated with increased FFM (2.8 +/- 0.5 kg), which was significantly greater than in the placebo group (P < 0.0001). Leg press strength increased significantly in testosterone-treated (P = 0.027), but not placebo treated, men; the difference between groups was not significant. Other measures of muscle performance and physical function did not change significantly in either group. Men receiving testosterone demonstrated significantly greater improvements in mental health and quality-of-life scores than those receiving placebo and improvements in fatigue/energy and mood scores that were not significantly different from those receiving placebo. Sexual function scores did not change in either group. In HIV-infected men with weight loss, a supraphysiological dose of testosterone significantly increased FFM but did not improve self-reported or performance-based measures of physical function. Improvements in mood, fatigue, and quality-of-life measures in the testosterone group, although clinically important, need further confirmation. PMID- 18430966 TI - Protein coingestion stimulates muscle protein synthesis during resistance-type exercise. AB - In contrast to the effect of nutritional intervention on postexercise muscle protein synthesis, little is known about the potential to modulate protein synthesis during exercise. This study investigates the effect of protein coingestion with carbohydrate on muscle protein synthesis during resistance-type exercise. Ten healthy males were studied in the evening after they consumed a standardized diet throughout the day. Subjects participated in two experiments in which they ingested either carbohydrate or carbohydrate with protein during a 2-h resistance exercise session. Subjects received a bolus of test drink before and every 15 min during exercise, providing 0.15 g x kg(-1) x h(-1) carbohydrate with (CHO + PRO) or without (CHO) 0.15 g x kg(-1) x h(-1) protein hydrolysate. Continuous intravenous infusions with l-[ring-(13)C(6)]phenylalanine and l-[ring (2)H(2)]tyrosine were applied, and blood and muscle biopsies were collected to assess whole body and muscle protein synthesis rates during exercise. Protein coingestion lowered whole body protein breakdown rates by 8.4 +/- 3.6% (P = 0.066), compared with the ingestion of carbohydrate only, and augmented protein oxidation and synthesis rates by 77 +/- 17 and 33 +/- 3%, respectively (P < 0.01). As a consequence, whole body net protein balance was negative in CHO, whereas a positive net balance was achieved after the CHO + PRO treatment (-4.4 +/- 0.3 vs. 16.3 +/- 0.4 micromol phenylalanine x kg(-1) x h(-1), respectively; P < 0.01). In accordance, mixed muscle protein fractional synthetic rate was 49 +/- 22% higher after protein coingestion (0.088 +/- 0.012 and 0.060 +/- 0.004%/h in CHO + PRO vs. CHO treatment, respectively; P < 0.05). We conclude that, even in a fed state, protein coingestion stimulates whole body and muscle protein synthesis rates during resistance-type exercise. PMID- 18430967 TI - Hormonal and purinergic stimulation of bicarbonate secretion in oviducts of rhesus monkey. AB - Because an increase in the HCO(3)(-) concentration of oviductal liquid at midcycle is believed to markedly enhance fertility, we have studied active secretion of HCO(3)(-) across highly differentiated cultures of monkey oviductal epithelium. Cultured cell sheets were mounted in Ussing chambers and bathed in medium containing 25 mM HCO(3)(-). Purinergic agents potently stimulated short circuit current (I(sc)) with an initial transient response declining within approximately 2 min to a sustained response. The potency sequence of ATP approximately UTP > ADP >> AMP suggested that the I(sc) response was mediated mainly by P2Y(2) receptors. Acetazolamide, an inhibitor of carbonic anhydrase, had little or no effect on baseline I(sc) or the transient response to ATP but abolished the sustained response to ATP. Similar results were obtained on sheets of native epithelium. In pH-stat experiments, the abluminal medium of cell cultures was bathed in HCO(3)(-)-CO(2) medium, and the pH of the unbuffered luminal medium was maintained at approximately 7.4 by addition of strong acid or base. ATP stimulated base secretion, and this was inhibited by acetazolamide. Furthermore, these changes in secretion of base were in good quantitative agreement with the I(sc) responses. When phenol red (an estrogen) was removed from the culture medium, ATP-dependent HCO(3)(-) secretion was markedly reduced but could be restored by treatment with estradiol. Estrogens also markedly increased ciliation of the cultures. These results suggest that the midcycle increase in the HCO(3)(-) concentration of oviductal liquid may be mediated by the effects of estradiol on purinergic pathways or on ATP secretion. PMID- 18430968 TI - Pyruvate dehydrogenase kinase-4 deficiency lowers blood glucose and improves glucose tolerance in diet-induced obese mice. AB - The effect of pyruvate dehydrogenase kinase-4 (PDK4) deficiency on glucose homeostasis was studied in mice fed a high-fat diet. Expression of PDK4 was greatly increased in skeletal muscle and diaphragm but not liver and kidney of wild-type mice fed the high-fat diet. Wild-type and PDK4(-/-) mice consumed similar amounts of the diet and became equally obese. Insulin resistance developed in both groups. Nevertheless, fasting blood glucose levels were lower, glucose tolerance was slightly improved, and insulin sensitivity was slightly greater in the PDK4(-/-) mice compared with wild-type mice. When the mice were killed in the fed state, the actual activity of the pyruvate dehydrogenase complex (PDC) was higher in the skeletal muscle and diaphragm but not in the liver and kidney of PDK4(-/-) mice compared with wild-type mice. When the mice were killed after overnight fasting, the actual PDC activity was higher only in the kidney of PDK4(-/-) mice compared with wild-type mice. The concentrations of gluconeogenic substrates were lower in the blood of PDK4(-/-) mice compared with wild-type mice, consistent with reduced formation in peripheral tissues. Diaphragms isolated from PDK4(-/-) mice oxidized glucose faster and fatty acids slower than diaphragms from wild-type mice. Fatty acid oxidation inhibited glucose oxidation by diaphragms from wild-type but not PDK4(-/-) mice. NEFA, ketone bodies, and branched-chain amino acids were elevated more in PDK4(-/-) mice, consistent with slower rates of oxidation. These findings show that PDK4 deficiency lowers blood glucose and slightly improves glucose tolerance and insulin sensitivity in mice with diet-induced obesity. PMID- 18430969 TI - Coordinated phosphorylation of insulin receptor substrate-1 by glycogen synthase kinase-3 and protein kinase C betaII in the diabetic fat tissue. AB - Serine/threonine phosphorylation of insulin receptor substrate-1 (IRS-1) is an important negative modulator of insulin signaling. Previously, we showed that glycogen synthase kinase-3 (GSK-3) phosphorylates IRS-1 at Ser(332). However, the fact that GSK-3 requires prephosphorylation of its substrates suggested that Ser(336) on IRS-1 was the "priming" site phosphorylated by an as yet unknown protein kinase. Here, we sought to identify this "priming kinase" and to examine the phosphorylation of IRS-1 at Ser(336) and Ser(332) in physiologically relevant animal models. Of several stimulators, only the PKC activator phorbol ester PMA enhanced IRS-1 phosphorylation at Ser(336). Treatment with selective PKC inhibitors prevented this PMA effect and suggested that a conventional PKC was the priming kinase. Overexpression of PKCalpha or PKCbetaII isoforms in cells enhanced IRS-1 phosphorylation at Ser(336) and Ser(332), and in vitro kinase assays verified that these two kinases directly phosphorylated IRS-1 at Ser(336). The expression level and activation state of PKCbetaII, but not PKCalpha, were remarkably elevated in the fat tissues of diabetic ob/ob mice and in high-fat diet-fed mice compared with that from lean animals. Elevated levels of PKCbetaII were also associated with enhanced phosphorylation of IRS-1 at Ser(336/332) and elevated activity of GSK-3beta. Finally, adenoviral mediated expression of PKCbetaII in adipocytes enhancedphosphorylation of IRS-1 at Ser(336). Taken together, our results suggest that IRS-1 is sequentially phosphorylated by PKCbetaII and GSK-3 at Ser(336) and Ser(332). Furthermore, these data provide evidence for the physiological relevance of these phosphorylation events in the pathogenesis of insulin resistance in fat tissue. PMID- 18430970 TI - Age-related changes in renal and hepatic cellular mechanisms associated with variations in rat serum thyroid hormone levels. AB - Aging is associated with changes in thyroid gland physiology. Age-related changes in the contribution of peripheral tissues to thyroid hormone serum levels have yet to be systematically assessed. Here, we investigated age-related alterations in the contributions of the liver and kidney to thyroid hormone homeostasis using 6-, 12-, and 24-mo-old male Wistar rats. A significant and progressive decline in plasma thyroxine occurred with age, but triiodothyronine (T(3)) was decreased only at 24 mo. This was associated with an unchanged protein level of the thyroid hormone transporter monocarboxylate transporter 8 (MCT8) in the kidney and with a decreased MCT8 level in the liver at 24 mo. Hepatic type I deiodinase (D1) protein level and activity declined progressively with age. Renal D1 levels were decreased at both 12 and 24 mo but D1 activity was decreased only at 24 mo. In the liver, no changes occurred in thyroid hormone receptor (TR) TRalpha(1), whereas a progressive increase in TRbeta(1) occurred at both mRNA and total protein levels. In the kidney, both TRalpha(1) and TRbeta(1) mRNA and total protein levels were unchanged between 6 and 12 mo but increased at 24 mo. Interestingly, nuclear TRbeta1 levels were decreased in both liver and kidney at 12 and 24 mo, whereas nuclear TRalpha(1) levels were unchanged. Collectively, our data show differential age-related changes among hepatic and renal MCT8 and D1 and TR expressions, and they suggest that renal D1 activity is maintained with age to compensate for the decrease in hepatic T(3) production. PMID- 18430971 TI - Agreement on intimate partner violence among a sample of blue-collar couples. AB - This study assessed agreement level about the occurrence of past-year male-to female partner violence (MFPV) and female-to-male partner violence (FMPV) among a sample of 897 blue-collar couples. Intimate partner violence (IPV) was measured with the Physical Assault subscale of the revised Conflict Tactics Scales (CTS2). Agreement level was assessed with Cohen's kappa statistic. Lower-bound estimates (based on couple agreement that an IPV event occurred) and upper-bound estimates (based on uncorroborated reports from either partner that an IPV event occurred) were calculated. Results indicated low agreement for most IPV behaviors (kappa < .40). Estimated lower- and upper-bound rates for MFPV were 6.7% and 21.2%, for FMPV, 7.1% and 24.2%, and for any IPV, 10.1% and 30.2%. Findings suggest that single-point IPV prevalence estimates are biased; lower- and upper-bound estimates using collateral reports should be calculated when possible. In addition, findings underscore the importance of conducting IPV research among understudied populations, such as working-class couples, that may be at elevated IPV risk. PMID- 18430972 TI - Identification of a novel GPCAT activity and a new pathway for phosphatidylcholine biosynthesis in S. cerevisiae. AB - Turnover of phospholipids in the yeast Saccharomyces cerevisiae generates intracellular glycerophosphocholine (GPC). Here we show that GPC can be reacylated in an acyl-CoA-dependent reaction by yeast microsomal membranes. The lysophosphatidylcholine that is formed in this reaction is efficiently further acylated to phosphatidylcholine (PC) by yeast microsomes, thus providing a new pathway for PC biosynthesis that can either recycle endogenously generated GPC or utilize externally provided GPC. Genetic and biochemical evidence suggests that this new enzymatic activity, which we call GPC acyltransferase (GPCAT), is not mediated by any of the previously known acyltransferases in yeast. The GPCAT activity has an apparent V(max) of 8.7 nmol/min/mg protein and an apparent K(m) of 2.5 mM. It has a neutral pH optimum, similar to yeast glycerol-3-phosphate acyltransferase, but differs from the latter in being more heat stable. The GPCAT activity is sensitive to N-ethylmaleimide, phenanthroline, and Zn(2+) ions. In vivo experiments showed that PC is efficiently labeled when yeast cells are fed with [(3)H]choline-GPC, and that this reaction occurs also in pct1 knockout strains, where de novo synthesis of PC by the CDP-choline pathway is blocked. This suggests that GPCAT can provide an alternative pathway for PC biosynthesis in vivo. PMID- 18430973 TI - Lipotoxicity: what is the fate of fatty acids? PMID- 18430974 TI - Sensorimotor rhythm-based brain-computer interface (BCI): model order selection for autoregressive spectral analysis. AB - People can learn to control EEG features consisting of sensorimotor rhythm amplitudes and can use this control to move a cursor in one or two dimensions to a target on a screen. Cursor movement depends on the estimate of the amplitudes of sensorimotor rhythms. Autoregressive models are often used to provide these estimates. The order of the autoregressive model has varied widely among studies. Through analyses of both simulated and actual EEG data, the present study examines the effects of model order on sensorimotor rhythm measurements and BCI performance. The results show that resolution of lower frequency signals requires higher model orders and that this requirement reflects the temporal span of the model coefficients. This is true for both simulated EEG data and actual EEG data during brain-computer interface (BCI) operation. Increasing model order, and decimating the signal were similarly effective in increasing spectral resolution. Furthermore, for BCI control of two-dimensional cursor movement, higher model orders produced better performance in each dimension and greater independence between horizontal and vertical movements. In sum, these results show that autoregressive model order selection is an important determinant of BCI performance and should be based on criteria that reflect system performance. PMID- 18430975 TI - Fully optimized discrimination of physiological responses to auditory stimuli. AB - The use of multivariate measurements to characterize brain activity (electrical, magnetic, optical) is widespread. The most common approaches to reduce the complexity of such observations include principal and independent component analyses (PCA and ICA), which are not well suited for discrimination tasks. We addressed two questions: first, how do the neurophysiological responses to elongated phonemes relate to tone and phoneme responses in normal children, and, second, how discriminable are these responses. We employed fully optimized linear discrimination analysis to maximally separate the multi-electrode responses to tones and phonemes, and classified the response to elongated phonemes. We find that discrimination between tones and phonemes is dependent upon responses from associative regions of the brain apparently distinct from the primary sensory cortices typically emphasized by PCA or ICA, and that the neuronal correlates corresponding to elongated phonemes are highly variable in normal children (about half respond with neural correlates of tones and half as phonemes). Our approach is made feasible by the increase in computational power of ordinary personal computers and has significant advantages for a wide range of neuronal imaging modalities. PMID- 18430977 TI - Naming of grammatical classes in frontotemporal dementias: linguistic and non linguistic factors contribute to noun-verb dissociation. AB - We studied noun and verb naming in three main variants of frontotemporal dementia: the frontal variant(Fv-FTD), primary progressive aphasia (PPA) and semantic dementia (SD). We further distinguished PPA in nonfluent and fluent forms and restricted diagnosis of SD to subjects with progressive semantic breakdown leading to agnosia for words and objects. Fv-FTD and nonfluent-PPA named objects better than actions, SD showed an inverse dissociation and no specific pattern emerged in fluent-PPA. In this last group, in spite of the broad definition of fluent aphasia, quite heterogeneous patterns of language disorders and word class dissociation emerged when single-subject analyses were performed. In fv-FTD correlations between executive tasks and action naming were stronger than between executive tasks and object naming. We conclude that both linguistic and non linguistic factors, in particular an executive deficit, contribute to grammatical class dissociation. We also suggest that the fluent vs. nonfluent distinction does not reflect the complexity of primary aphasia. PMID- 18430976 TI - Improved bladder emptying in urinary retention by electrical stimulation of pudendal afferents. AB - Urinary retention is the inability to empty the bladder completely, and may result from bladder hypocontractility, increases in outlet resistance or both. Chronic urinary retention can lead to several urological complications and is often refractory to pharmacologic, behavioral and surgical treatments. We sought to determine whether electrical stimulation of sensory fibers in the pudendal nerve could engage an augmenting reflex and thereby improve bladder emptying in an animal model of urinary retention. We measured the efficiency of bladder emptying with and without concomitant electrical stimulation of pudendal nerve afferents in urethane-anesthetized rats. Voiding efficiency (VE = voided volume/initial volume) was reduced from 72 +/- 7% to 29 +/- 7% following unilateral transection of the sensory branch of the pudendal nerve (UST) and from 70 +/- 5% to 18 +/- 4% following bilateral transection (BST). Unilateral electrical stimulation of the proximal transected sensory pudendal nerve during distention-evoked voiding contractions significantly improved VE. Low-intensity stimulation at frequencies of 1-50 Hz increased VE to 40-51% following UST and to 39-49% following BST, while high-intensity stimulation was ineffective at increasing VE. The increase in VE was mediated by increases in the duration of distention-evoked voiding bladder contractions, rather than increases in contraction amplitude. These results are consistent with an essential role for pudendal sensory feedback in efficient bladder emptying, and raise the possibility that electrical activation of pudendal nerve afferents may provide a new approach to restore efficient bladder emptying in persons with urinary retention. PMID- 18430978 TI - The diagnostic role of serum inflammatory and soluble proteins on dementia subtypes: correlation with cognitive and functional decline. AB - In the past years, the possible involvement of inflammation in the pathogenesis of dementia has been the subject of several investigations. However there are restricted data about the profile of the inflammatory and soluble proteins in well evaluated Alzheimer's disease (AD), vascular dementia (VD), mild cognitive impairment (MCI) and healthy controls. There are also no reliable data regarding the relationship between the overlapping protein levels and cognitive or functional decline. We measured levels of IL-1beta, IL-2, IL-6, IL-18, TNF-alpha, beta-Amlyloid 1-40 and alpha1-antichymotrypsin levels in plasma in groups of total 82 subjects with AD, MCI, VD and controls using enzyme-linked immunosorbent assay (ELISA) method. Our study samples showed high levels of proinflammatory cytokine levels (especially IL-18) in all patient groups but only high levels of alpha1-antichymotrypsine in VD patients compared to controls. There is no significant correlation between the laboratory and clinical variables except for a link between IL-1beta and NPI scores of AD. In conclusion, this study yielded evidence of some shared mechanisms underlying AD and VD and thus motivates further studies of inflammatory markers in various types of dementia and MCI. PMID- 18430979 TI - The potential of dyslexic individuals in communication design education. AB - If dyslexic individuals have the ability to express themselves in different ways, particularly in the field of modern graphic design, would they be a favoured group in creating the extraordinary and outstanding ideas that are required in communication design? The study group consisted of 20 primary school dyslexics between ages of 7-12 and 20 non-dyslexics serving as a control group. A jury with four specialists evaluated the drawings gathered from the 40 participants. Even though we might not say surely that the dyslexics are the best possible candidates for communication design education, based on the statistical results we have concluded that they should be among the potential candidates for both general communication design education and for more specific minor study areas such as icon design. PMID- 18430980 TI - Neuropsychological rehabilitation in mild and moderate Alzheimer's disease patients. AB - OBJECTIVE: The purpose of this study was to analyze the effect of a neuropsychological rehabilitation (NR) program on patients with Alzheimer's disease (AD). METHODS: The sample was composed of 16 elderly outpatients who participated in an open trial with rivastigmine (6 to 12 mg/day) for 4 months and were randomized to 3 different groups: 1. group NR (N=5), 2. individualized NR (N=6) and 3. NR at home under supervision of a relative or caregiver (N=5). All 3 groups fulfilled the same NR protocol consisting of a once a week session. Just before and after the 22 week period of rehabilitation, all patients were evaluated using psychiatric and functional scales, and neuropsychological tests by interviewers that did not participate in the cognitive training. RESULTS: The intervention did not produce any statistically significant change, but small gains were observed on some cognition tests, activities of daily living (ADL), and psychiatric symptoms in groups 1 and 2. CONCLUSION: Group NR is recommended for reducing psychiatric symptoms, and individualized NR for improving ADL. NR at home either has no associated benefits, or the training sessions were not appropriately conducted by the caregiver. However, additional research with larger samples is necessary to confirm these observations. PMID- 18430981 TI - Spatial perseveration in dementia with Lewy bodies. PMID- 18430982 TI - Severe Broca's aphasia without Broca's area damage. PMID- 18430983 TI - Right hemisphere involvement in non-fluent primary progressive aphasia. AB - We described a 56-years-old man with a diagnosis of "non-fluent primary progressive aphasia" (NfPPA). An accurate neuropsychological, neurological and neuroimaging evaluation was performed in order to assess clinical and behavioural features of the patient. From a neuropsychological point of view, the patient showed a typical cognitive profile of subjects affected by NfPPA: a prominent language deficit, associated with impairments in several cognitive domains after three years from the onset of the symptomatology. The most intriguing feature is that SPECT revealed hypoperfusion in the right frontal cortex, albeit the patient is right-handed. This unexpected finding shows that NfPPA may arise not only from cortical abnormalities in the language-dominant left hemisphere, but also from right hemisphere involvement in a right hander (crossed aphasia). PMID- 18430984 TI - Effects of decompressive surgery on prognosis and cognitive deficits in herpes simplex encephalitis. AB - Herpes simplex encephalitis (HSE) is a serious viral infection with a high rate of mortality. The most commonly seen complications are behavioral changes, seizures and memory deficits. We report the case of a 37-year-old man with HSE in the right temporal lobe and a severe midline shift who was treated with acyclovir. The patient underwent anterior temporal lobe resection. Although HSE can cause permanent cognitive deficits, in this case, early surgical intervention minimized any deficit, as determined by detailed neuropsychological examination. Surgical decompression is indicated as early as possible in severe cases. This case report emphasizes the effect of surgical decompression for HSE on cognitive function, which has rarely been mentioned before. PMID- 18430985 TI - An in silico analysis of cytochrome c from Phanerochaete chrysosporium: its amino acid sequence and characterization of gene structural elements. AB - An in silico approach was used to investigate cytochrome c and the cytochrome c gene of Phanerochaete chrysosporium. The cytochrome c gene contains four introns. Omission of the introns reveals a DNA sequence coding for a complete predicted amino acid sequence for P. chrysosporium cytochrome c consistent with those of other cytochromes c. Fungal cytochromes c often have a short N-terminal peptide preceding a Gly that is the N-terminal amino acid in many cytochromes c. Thus a microexon codes for an N-terminal pentapeptide (MetProTyrAlaPro) in P. chrysosporium that is identical to the N-terminal pentapeptide of Schizosaccharomyces pombe, a well studied yeast, the genome of which bears more similarity to higher eukaryotes than to other fungi. The fourth intron, when omitted, reveals the presence of another microexon resulting in a sequence for the C-terminal portion of the protein and the stop codon. Interestingly, two interpretations for the sequence of this intron leads to predictions that the C terminal sequence ends with either AlaValAsn or AlaTyr. Selected aspects of the molecular architecture of cytochrome c and regulatory control elements of the P. chrysosporium cytochrome c gene were analyzed and compared to those present in other fungi and to those present in genes for lignin peroxidases and cytochromes P-450, two important families of hemeproteins produced by this fungus. PMID- 18430986 TI - Evolutionary origin of the protozoan parasites histone-like proteins (HU). AB - The histone-like proteins (HU) belong to a family of DNA architectural proteins that stabilize nucleoprotein complexes. We found a putative HU protein (TgGlmHMM_3045) in Toxoplasma gondii genome that was homologous to the bacterial HU protein. This putative sequence was located in the scaffold TGG_995361 of the chromosome 10. The sequence included the prokaryotic bacterial histone-like domain, KFGSLGlRRRGERVARNPRT (ID number PS00045). HU protein sequences were also found in Plasmodium falciparum, Neospora caninum, Theileria parva and Theileria annulata. We found that the homology of the putative HU protein in Apicomplexa was greater with bacterial histone-like proteins than with eukaryotic histone proteins. The phylogenetic tree indicated that the putative HU protein genes were acquired in Apicomplexa by means of a secondary endosymbiotic event from red algae and later they were transferred from the apicoplast organelle to the nuclear genome. PMID- 18430987 TI - The GTP binding sites interacted with RNA-dependent RNA polymerase of classical swine fever virus in de novo initiation. AB - The NS5B protein of classical swine fever virus (CSFV) is an important enzyme bearing a unique RNA-dependent RNA polymerase (RdRp) activity. The RdRp plays a crucial role in the viral replication cycle and in forming a replicase complex. However, the initiating synthesis mechanism of the CSFV RNA polymerase is unclearly described at present. Our aim is to reveal the RdRp-GTP docking sites and the effective modules of GTP initially bound to the polymerase in starting initiation of replication according to a well predicted CSFV RdRp model. Based on some known crystal structures of RNA polymerase, computational methods were used to establish the model of a CSFV RdRp. An analogous mechanism of CSFV RNA polymerase in de novo initiation was subsequently represented through docking a GTP into the structure model. The unique GTP binding pocket of the polymerase was pointed out: five residues E227, S408, R427, K435, and R439 involved in steady hydrogen bonds and two residues C407 and L232 involved in hydrophobic contact with the GTP. From a genetic evolutionary point of view, three residues C407, S408 and R427 have been suggested to be of particular importance by analysis of residue conservation. It is suggested that these crucial residues should have very significant function in the de novo initiation of the rigorous CSFV polymerase model, which can lead us to design experiments for studying the mechanism of viral replication and develop valid anti-viral drugs. PMID- 18430988 TI - Distinct patterns in the regulation and evolution of human cancer genes. AB - Understanding the mechanism of regulation of cancer genes and the constraints on their coding sequences is of fundamental importance in understanding the process of tumour development. Here we test the hypothesis that tumour suppressor genes and proto-oncogenes, due to their involvement in tumourigenesis, have distinct patterns of regulation and coding selective constraints compared to non-cancer genes. Indeed, we found significantly greater conservation in the promoter regions of proto-oncogenes, suggesting that these genes are more tightly regulated, i.e. they are more likely to contain a higher density of cis regulatory elements. Furthermore, proto-oncogenes appear to be preferentially targeted by microRNAs and have longer 3' UTRs. In addition, proto-oncogene evolution appears to be highly constrained, compared to tumour suppressor genes and non-cancer genes. A number of these trends are confirmed in breast and colon cancer gene sets recently identified by mutational screening. PMID- 18430989 TI - NetAtlas: a Cytoscape plugin to examine signaling networks based on tissue gene expression. AB - Graphical methods are useful for visualizing signaling networks derived from the synthesis of large bodies of literature information or large-scale experimental measurements. Software tools to filter and organize these networks allow the exploration of their inherent biological and structural properties. We have developed NetAtlas, an open-source, Java-based Cytoscape plugin for examining signaling networks in the context of tissue gene expression patterns. The tissue gene expression data available through NetAtlas consists of 79 human tissues, 61 mouse tissues, and 44 combined tissues from 3 rat strains. Users may also import their own tissue gene expression data. The NetAtlas plugin allows the creation of tissue-defined signaling networks by identifying which components are expressed in particular tissues, which components show tissue-specific expression, and which components within the network are coordinately expressed across tissues. The NetAtlas plugin is available at http://sourceforge.net/projects/netatlas/. PMID- 18430990 TI - BioCompass: a novel functional inference tool that utilizes MeSH hierarchy to analyze groups of genes. AB - Microarray technology has become employed widely for biological researchers to identify genes associated with conditions such as diseases and drugs. To date, many methods have been developed to analyze data covering a large number of genes, but they focus only on statistical significance and cannot decipher the data with biological concepts. Gene Ontology (GO) is utilized to understand the data with biological interpretation; however, it is restricted to specific ontology such as biological process, molecular function, and cellular component. Here, we attempted to apply MeSH (Medical Subject Headings) to interpret groups of genes from biological viewpoint. To assign MeSH terms to genes, in this study, contexts associated with genes are retrieved from full set of MEDLINE data using machine learning, and then extracted MeSH terms from retrieved articles. Utilizing the developed method, we implemented a software called BioCompass. It generates high-scoring lists and hierarchical lists for diseases MeSH terms associated with groups of genes to utilize MeSH and GO tree, and illustrated a wiring diagram by linking genes with extracted association from articles. Researchers can easily retrieve genes and keywords of interest, such as diseases and drugs, associated with groups of genes. Using retrieved MeSH terms and OMIM in conjunction with, we could obtain more disease information associated with target gene. BioCompass helps researchers to interpret groups of genes such as microarray data from a biological viewpoint. PMID- 18430991 TI - M@IA: a modular open-source application for microarray workflow and integrative datamining. AB - Microarray technology is a widely used approach to gene expression analysis. Many tools for microarray management and data analysis have been developed, and recently new methods have been proposed for deciphering biological pathways by integrating microarray data with other data sources. However, to improve microarray analysis and provide meaningful gene interaction networks, integrated software solutions are still needed. Therefore, we developed M@IA, an environment for DNA microarray data analysis allowing gene network reconstruction. M@IA is a microarray integrated application which includes all of the steps of a microarray study, from MIAME-compliant raw data storage and processing gene expression analysis. Furthermore, M@IA allows automatic gene annotation based on ontology, metabolic/signalling pathways, protein interaction, miRNA and transcriptional factor associations, as well as integrative analysis of gene interaction networks. Statistical and graphical methods facilitate analysis, yielding new hypotheses on gene expression data. To illustrate our approach, we applied M@IA modules to microarray data taken from an experiment on liver tissue. We integrated differentially expressed genes with additional biological information, thus identifying new molecular interaction networks that are associated with fibrogenesis. M@IA is a new application for microarray management and data analysis, offering functional insights into microarray data by the combination of gene expression data and biological knowledge annotation based on interactive graphs. M@IA is an interactive multi-user interface based on a flexible modular architecture and it is freely available for academic users at http://maia.genouest.org. PMID- 18430992 TI - An integrated and unifying hypothesis for the metabolic basis of sporadic Alzheimer's disease. AB - Acquired disturbances of several aspects of cellular metabolism appear pathologically important in sporadic Alzheimer's disease (SAD). Among these, brain glucose utilization is reduced in the early stages of the disease. Hyperinsulinemia, which is a characteristic finding of insulin resistance, results in a central insulin deficit. Insufficient insulin signaling impairs the intricate balance of nitric oxide regulation of the central nervous system. Reduction in central insulin decreases neuronal nitric oxide synthase and increases inducible synthase activity. This, in turn, decreases astrocytic energy substrates and antioxidant supply of neurons. In addition, an increase in peroxynitrite formation impairs redox balance. Hyperleptinemia and glucose excess, which are the other parameters of insulin resistance, may worsen the reduced astrocytic energy supply and the ongoing inflammation via the inhibition of AMP-activated protein kinase (AMPK). Consequently, energy deficit and inflammation in neuronal tissue may cause neurodegeneration of SAD. PMID- 18430994 TI - Gonadotropins and cognition in older women. AB - Recent research studies associate elevated gonadotropin levels with dementia. Specifically, an age associated increase in levels of luteinizing hormone has been linked to an increased risk of Alzheimer's disease. The objective of this study was to investigate the association between gonadotropin levels and cognition in older, healthy postmenopausal women. Cognitive functioning was compared with plasma levels of estradiol, luteinizing hormone, follicle stimulating hormone, Abeta40 and APOE genetic status in 649 community-dwelling, non-demented older women residing in Western Australia. High endogenous luteinizing hormone levels were associated with a lower cognitive score, especially in older women and in those women that were depressed. Unexpectedly, disproportionately well preserved cognitive functioning was found for the oldest women who had high endogenous levels of follicle stimulating hormone. The findings indicate that gonadotropins can impact upon cognitive functioning in older postmenopausal women, and that luteinizing hormone and follicle stimulating hormone may exert contrasting effects. Taken together, the findings have important implications for the development of possible preventive strategies for dementia. PMID- 18430993 TI - Multiple SNPs within and surrounding the apolipoprotein E gene influence cerebrospinal fluid apolipoprotein E protein levels. AB - The epsilon4 allele of the apolipoprotein E gene (APOE) is associated with increased risk and earlier age at onset in late onset Alzheimer's disease (AD). Other factors, such as expression level of apolipoprotein E protein (apoE), have been postulated to modify the APOE related risk of developing AD. Multiple loci in and outside of APOE are associated with a high risk of AD. The aim of this exploratory hypothesis generating investigation was to determine if some of these loci predict cerebrospinal fluid (CSF) apoE levels in healthy non-demented subjects. CSF apoE levels were measured from healthy non-demented subjects 21-87 years of age (n=134). Backward regression models were used to evaluate the influence of 21 SNPs, within and surrounding APOE, on CSF apoE levels while taking into account age, gender, APOE epsilon4 and correlation between SNPs (linkage disequilibrium). APOE epsilon4 genotype does not predict CSF apoE levels. Three SNPs within the TOMM40 gene, one APOE promoter SNP and two SNPs within distal APOE enhancer elements (ME1 and BCR) predict CSF apoE levels. Further investigation of the genetic influence of these loci on apoE expression levels in the central nervous system is likely to provide new insight into apoE regulation as well as AD pathogenesis. PMID- 18430995 TI - Mitochondrial transcription factor A (TFAM) gene variation and risk of late-onset Alzheimer's disease. AB - Impaired mitochondrial function and an increased number of mutations in mitochondrial DNA (mtDNA) has been found in brains of patients with late-onset Alzheimer's disease (LOAD). The TFAM-gene encodes the mitochondrial transcription factor A, a protein that controls the transcription, replication, damage sensing, and repair of mtDNA. TFAM is on human chromosome region 10q21.1, where a locus for LOAD has been mapped. Our objective was to determine the role of TFAM-gene variation in the risk of LOAD. The seven TFAM coding exons were analysed through single strand conformation analysis and direct sequencing in a cohort of Spanish LOAD-patients and healthy controls. We found four common polymorphisms, two in the flanquing intronic and two in the coding sequences. Polymorphism rs1937 (+35 G/C) was the only missense change (S12T). Genotyping of this polymorphism in 300 LOAD-patients and 183 healthy controls showed a significantly higher frequency of GG-homozygotes in the patients (92% vs. 86%; p=0.04; OR=1.91, 95%CI=1.02-3.50). This suggests that S12 is a risk factor for LOAD in our population. In conclusion, rare variants (mutations) in the TFAM gene were not found in LOAD patients, but the S12T polymorphism was a moderate risk factor for LOAD in our population. PMID- 18430996 TI - Inhibition of PHF-like tau hyperphosphorylation in SH-SY5Y cells and rat brain slices by K252a. AB - Abnormal hyperphosphorylation of tau is believed to constitute a critical biochemical event in the process of neurofibrillary degeneration of Alzheimer's disease. We have developed a cellular model where apparently authentic PHF-like tau hyperphosphorylation is induced by okadaic acid. To gain deeper insight into the complex mechanisms of this pathological process we tested a variety of kinase inhibitors in this model. We found that K252a is differentiated from staurosporine by its inhibition of ERK2: both compounds are structurally related microbial metabolites generally believed to have only moderate kinase selectivity. However, since ERK2 inhibitors are exceedingly rare, we used this differential inhibitory property of K252a to demonstrate the involvement of ERK2 in PHF-type tau hyperphosphorylation. K252a was uniquely able to completely suppress the okadaic acid-induced tau hyperphosphorylation in SH-SY5Y cells and rat brain slices by way of including ERK2 in its inhibitory spectrum, and to conserve the normal binding of tau to tubulin. GSK3 inhibitors partially affected the normal state of tau phosphorylation in SH-SY5Y cells, but had no impact on okadaic acid-induced tau hyperhosphorylation. As K252a is the first molecule identified capable of preventing the spectrum of PHF-like tau hyperphosphorylation markers, it may represent a conceptual starting point for therapeutic development of suitable spectrum kinase inhibitors. PMID- 18430997 TI - Decrease of protein phosphatase 2A and its association with accumulation and hyperphosphorylation of tau in Down syndrome. AB - Virtually all individuals with Down syndrome (DS) develop neurofibrillary tangles, a characteristic brain lesion of Alzheimer's disease (AD), when they reach the fourth decade of life. In AD, neurofibrillary tangles are thought to result from abnormal hyperphosphorylation of tau protein, which, in turn, can result from down-regulation of protein phosphatase (PP) 2A, a major brain tau phosphatase. The abnormal hyperphosphorylation of tau in DS had not yet been characterized, and its causes were not understood. In this study, by using quantitative Western blot analysis, we found that the level of the catalytic subunit of PP2A, but not of PP1, PP2B or PP5, was dramatically decreased. The decrease of PP2A level correlated negatively to tau level and tau phosphorylation at several abnormal hyperphosphorylation sites, including Ser199, Thr205, Thr212, Ser262, Ser396 and Ser422. Our results indicate that PP2A is down-regulated in DS brain and suggest that this down-regulation might be involved in the abnormal hyperphosphorylation and accumulation of tau. PMID- 18430998 TI - Colostrinin: an oxidative stress modulator for prevention and treatment of age related disorders. AB - Colostrum-derived proline-rich polypeptide, also known as Colostrinin (CLN), has been shown to have a stabilizing effect on cognitive function in Alzheimer's disease patients. This complex action of CLN could be related to prevention of amyloid-beta peptide aggregation, as shown in in vitro studies, and its impact on delicate cassettes of signaling pathways common to cellular redox regulation, proliferation and differentiation. Studies on cultured cells showed that CLN modulates intracellular levels of reactive oxygen species (ROS), via regulation of glutathione metabolism, activity of antioxidant enzymes and mitochondria function. Due to an improvement in senescence-associated mitochondrial dysfunction and a decrease in ROS generation, CLN decelerates the aging processes of both cultured cells and experimental animals. When given orally to mice, CLN increased the lifespan and improved various motor and sensory activities. Although the molecular basis by which CLN exerts its diverse effects are still under investigation, the regulatory effect on the cellular redox state via maintenance of mitochondrial function and modification of ROS-induced cell signaling seem to be of great importance. In this article, we examine experimental data pertinent to the mechanism of action, including a review of CLN's utility in the maintenance of physiological processes in which oxidative stress has an etiological role. PMID- 18431000 TI - Haplotypes across ACE and the risk of Alzheimer's disease: the three-city study. AB - The purpose of this study was to examine the impact of two polymorphisms (rs4291A>T and rs4343G>A) in the ACE gene on the risk of Alzheimer's disease (AD), using a population-based cohort of 9294 subjects selected from the electoral rolls of three French cities (the Three-City Study). Two follow-up examinations took place 2 and 4 years after inclusion. Diagnosis of dementia was assessed at baseline and at each follow-up examination by neurologists independent of the 3C Study group. For the present analysis, subjects whose mother tongue was not French, those from abroad and those lost at follow-up were excluded, leaving a sample of 6791 subjects. 108 subjects were demented at baseline and 216 subjects, among which 141 had AD, developed a dementia during follow-up. The genotype distributions of the ACE SNPs rs4291 and rs4343 did not differ according to cognitive status. After adjustment for confounding variables, the risk of developing AD was similar whatever the genotype (rs4291 AT vs TT: OR=0.90, p=0.65; AA vs TT: OR=1.05, p= 0.84; rs4343 GA vs GG: OR=1.15, p= 0.48; AA vs GG: OR=1.25, p= 0.37). No global haplotype effect could be observed on the risk of AD. PMID- 18430999 TI - Intranasal insulin administration dose-dependently modulates verbal memory and plasma amyloid-beta in memory-impaired older adults. AB - Intranasal insulin administration raises central nervous system (CNS) insulin levels in humans and acutely facilitates verbal memory in patients with Alzheimer's disease (AD), an effect that may differ by APOE genotype. The purpose of this study was to examine the cognitive dose response curves for intranasal insulin administration, and determine whether the effects of insulin differ between participants with (epsilon4+) and without (epsilon4-) the APOE- epsilon4 allele. On separate mornings, 33 memory-impaired adults with AD or amnestic mild cognitive impairment and 59 normal adults each underwent five intranasal treatment conditions consisting of insulin (10, 20, 40, or 60 IU) or placebo. Cognition was tested 15-minutes post-treatment, and blood was acquired at baseline and 45-minutes post-treatment. Plasma insulin and glucose levels were unaffected by treatment. Insulin administration facilitated recall on two measures of verbal memory in memory-impaired epsilon4- adults, with performance generally peaking at 20 IU. In contrast, memory-impaired epsilon4+ subjects demonstrated a relative decline in verbal memory. Insulin also differentially modulated plasma amyloid-beta for memory-impaired subjects and normal controls, effects that again differed by APOE genotype. These findings suggest that groups with different genetic risks for AD may show differential dose-response curves following intranasal insulin administration. PMID- 18431001 TI - Gotu Kola (Centella Asiatica) extract enhances phosphorylation of cyclic AMP response element binding protein in neuroblastoma cells expressing amyloid beta peptide. AB - Alzheimer's disease (AD) is a progressive neurodegenerative disorder that shows cognitive deficits and memory impairment. Extract from the leaves of Gotu Kola (Centella Asiatica) have been used as an alternative medicine for memory improvement in Indian Ayurvedic system of medicine for a long time. Although several studies have revealed its effect in ameliorating the cognitive impairment in rat models of AD and stimulating property on neuronal dendrites of hippocampal region, the molecular mechanism of Gotu Kola on neuroprotection still remains to be elucidated. In this study, we report that phosphorylation of cyclic AMP response element binding protein (CREB) is enhanced in both a neuroblastoma cell line expressing amyloid beta 1-42 (Abeta) and in rat embryonic cortical primary cell culture. In addition, the contribution of two major single components to the enhanced CREB phosphorylatioin was examined. Furthermore, inhibitors were applied in this study revealing that ERK/RSK signaling pathway might mediate this effect of Gotu Kola extract. Taken together, we provide a possible molecular mechanism for memory enhancing property of Gotu Kola extract for the first time. PMID- 18431002 TI - Acute neglect rehabilitation using repetitive prism adaptation: a randomized placebo-controlled trial. AB - PURPOSE: At present, prism adaptation is probably the most promising rehabilitation procedure for hemi-neglect. However, randomised controlled trials are lacking and no data are available on the effectiveness of prism adaptation in the treatment of acute neglect. METHODS: We followed sixteen neglect patients using a randomised controlled design in which six patients received four-day-in-a row placebo treatment (CG) and ten patients received four-day-in-a row experimental treatment with 10 degrees rightward deviating prisms (EG) during their stay on the stroke unit. We examined whether patients in the EG improved faster than the CG by administering three neglect tasks (Schenkenberg Line Bisection, Letter Cancellation, Gainotti Scene Copying) immediately before and after each treatment. Second, we examined whether patients in the EG demonstrated a better long-term outcome at one month post-treatment (Behavioural Inattention Test). RESULTS: Patients in the EG improved faster on spatial tasks (line bisection, cancellation) than the CG but not on visuo-construction. Patients in the EG showed no differences with the CG in neglect outcome at one month post treatment. CONCLUSIONS: Four consecutive prism sessions produced beneficial effects in patients with acute neglect. However, prism effects were either short term, or placebo treatment with repeated pointing and/or repeated neglect testing was more helpful than we anticipated. Our results emphasize the importance of a placebo condition and a follow-up in rehabilitation studies. PMID- 18431003 TI - MRI infarction load and CI therapy outcomes for chronic post-stroke hemiparesis. AB - PURPOSE: To evaluate the effect of total cerebral infarction volume on motor status and treatment responses to Constraint-Induced Movement therapy (CI therapy) in patients with chronic post-stroke hemiparesis and limb nonuse. In most studies of clinical-radiological correspondences, infarct volume has been found to predict clinical status. METHODS: 60 adult patients who met inclusion criteria for CI therapy underwent standard structural brain MRI around the time of treatment. We tested whether the total volume of infarcted tissue predicted pre-treatment values and pre- to post-treatment changes on standard assessments of laboratory- and real-world-based upper extremity movement. RESULTS: The patients significantly improved from baseline clinical motor measures after CI therapy. The range of infarction volumes was comparable to those of other stroke studies. In contrast, total infarction volume was not correlated with any of the motor measures. The absence of correlation was maintained even when evaluating only patients with either unilateral or solitary cerebral infarctions, or with respect to severity of baseline motor impairment or variations in the forms of therapy. CONCLUSION: The findings suggest that certain standard motor measures in chronic hemiparetic stroke (limb movement in the laboratory, limb use in the life situation, changes in these measures following efficacious CI therapy) are insensitive to infarction load. Our findings are consistent with the hypothesis that plastic brain reorganization after stroke attenuates the effect of infarct volume on purposive limb movement. However, we cannot exclude the possibilities that the specific methods we used to assess upper limb status may be generally insensitive to infarction load, or that the stringent inclusion criteria for this research precluded detecting an effect of infarction load. Further study to evaluate these relationships in the acute-subacute stroke phases and among patients with more severe motor deficits would help to clarify the basis for the lack of an infarction load effect. PMID- 18431004 TI - Effects of behaviorally active ACTH (4-10) analogue - Semax on rat basal forebrain cholinergic neurons. AB - PURPOSE: It is well established that cholinergic neurons of the basal forebrain degenerate in Alzheimer's dementia. Although recent studies were concentrated on screening molecules that might reduce the concomitant cell loss, little is known about therapeutically promising molecules. We studied the effect of Semax (Met Glu-His-Phe-Pro-Gly-Pro), a behaviorally active adrenocorticotropic hormone (4 10) analogue, on survival of cholinergic basal forebrain neurons in vitro. Semax is known to stimulate learning and memory and can be successfully used for treatment of ischemic stroke. METHODS: Primary cultures of neuronal and glial cells from basal forebrain of rats were used in all experiments. The stability of Semax in cell cultures was tested by HPLC analysis. Cell survival in neuronal cultures was quantitated using immocytochemical and cytochemical analyses as well as detection of choline acetyltransferase activity. RESULTS: We have shown that Semax may approximately 1.5-1.7 fold increase survival of cholinergic basal forebrain neurons in vitro. Moreover, Semax (100 nM) stimulated activity of choline acetyltransferase in dissociated basal forebrain tissue cultures. However, the numbers of GABA-ergic neurons, total neuron specific enolase neurons were not affected. In concentration from 1 nM to 10 microM, Semax did not affect proliferation of glial cells in primary cultures. CONCLUSION: Implications of these findings with respect to Alzheimer's disease remain to be clarified. PMID- 18431005 TI - TrkB gene transfer does not alter hippocampal neuronal loss and cognitive deficits following traumatic brain injury in mice. AB - PURPOSE: The ability of brain-derived neurotrophic factor (BDNF) to attenuate secondary damage and influence behavioral outcome after experimental traumatic brain injury (TBI) remains controversial. Because TBI can result in decreased expression of the trkB receptor, thereby preventing BDNF from exerting potential neuroprotective effects, the contribution of both BDNF and its receptor trkB to hippocampal neuronal loss and cognitive dysfunction were evaluated. METHODS: Full length trkB was overexpressed in the left hippocampus of adult C57Bl/6 mice using recombinant adeno-associated virus serotype 2/5 (rAAV 2/5). EGFP (enhanced green fluorescent protein) expression was present at two weeks after AAV-EGFP injection and remained sustained up to four weeks after the injection. At 2 weeks following gene transduction, mice were subjected to parasagittal controlled cortical impact (CCI) brain injury, followed by either BDNF or PBS infusion into the hippocampus. RESULTS: No differences were observed in learning ability at two weeks post injury or in motor function from 48 hours to two weeks among treatment groups. The number of surviving pyramidal neurons in the CA2-CA3 region of the hippocampus was also not different among treatment groups. CONCLUSIONS: These data suggest that neither overexpression of trkB, BNDF infusion or their combination affects neuronal survival or behavioral outcome following experimental TBI in mice. PMID- 18431007 TI - Surveillance of Indians with liver cirrhosis for treatable hepatocellular carcinoma: another enigma. PMID- 18431006 TI - Overexpression of cellular prion protein alters postischemic Erk1/2 phosphorylation but not Akt phosphorylation and protects against focal cerebral ischemia. AB - PURPOSE: The physiological function of the cellular prion protein (PrPC) is still unclear. A growing body of evidence suggests that PrPC has neuroprotective properties and that its deletion increases susceptibility to focal cerebral ischemia. The purpose of this study was to elucidate the role of PrPC overexpression in ischemic brain injury in vivo. METHODS: PrPC overexpressing (TG35) and wild type (WT) mice were subjected to a 90-minute transient focal cerebral ischemia followed by infarct volume analysis 24 hours after lesion. To identify effects of PrPC overexpression on signalling pathways important for the regulation of ischemic cell death, we studied postischemic activation and expression of Akt and Erk1/2 using quantitative Western Blot analysis. RESULTS: TG35 mice displayed significantly smaller infarct volumes and showed reduced early postischemic Erk1/2 phosphorylation, a pathway known to exacerbate neuronal injury following transient cerebral ischemia. In contrast, PrPC overexpression did not change postischemic Akt phosphorylation, which acts anti-apoptotic and is reduced in PrPC knockout animals. CONCLUSIONS: These results demonstrate that PrPC overexpression reduces deleterious Erk1/2 activation but does not affect Akt activation after transient cerebral ischemia, suggesting a role for distinct cytosolic signalling pathways in PrPC mediated neuroprotection. PMID- 18431008 TI - Risk factors for esophageal cancer in Serbia. AB - AIM: To establish the role of specified risk factors in the etiology of squamous cell carcinoma of esophagus in Serbia. METHODS: The hospital based case-control study included 102 newly diagnosed patients with squamous cell esophageal cancer and the same number of age and sex matched hospital controls. In the analysis of data conditional univariate and multivariate logistic regressions were applied. RESULTS: The factors significantly related to cancer of esophagus were: smoking (OR=2.0, 95%CI 1.0-4.2), daily consumption of hard liquor (OR=7.5, 95%CI 2.8 20.0), low educational level (OR=4.7, 95%CI 2.2-10.0), occupational exposure to chemicals (OR=3.7, 95%CI 1.6-9.0), malignant tumors in family history (OR=3.6, 95%CI 1.6-9.0), consumption of wine (OR=0.1, 95%CI 0.1-0.3,) and non-carbonated beverages (OR=0.3, 95%CI 0.2-0.7). CONCLUSIONS: Hard liquor and tobacco appear to be the important risk factors for development of squamous cell esophageal cancer in Serbia, whereas wine and non-carbonated beverages may play a protective role. PMID- 18431009 TI - Time to recognize atypical celiac disease in Indian children. AB - BACKGROUND/OBJECTIVE: There is scant information about atypical (non-diarrheal) presentation of celiac disease (CD) from India. We conducted this study to compare non-diarrheal and diarrheal presentations of CD in children. METHODS: From November 2003 to December 2005, we prospectively screened two groups of children for CD, group I with diarrhea and group II without diarrhea but with atypical presentations (unexplained growth retardation, refractory anemia, refractory rickets, chronic constipation and abdominal distension). Screening was done with IgA antiendomysial antibody (EMA) followed by duodenal biopsy if EMA was positive. Celiac disease was diagnosed according to modified ESPGHAN criteria. RESULTS: A total of 200 children were screened (103 in group I and 97 in group II) and CD was diagnosed in 42 (classical 24, atypical 18). Presentation of atypical CD were; short stature 6, anemia 4, abdominal distension 3, rickets 2, and constipation, diabetes mellitus, delayed puberty in 1 case each. Patients with atypical CD were older (median age 10.4 years vs 5.5 years, p< 0.007) than classical cases. On mean (SD) follow-up of 12.6 (7.5) months all showed response to gluten-free-diet, and median gain in weight, height and final hemoglobin levels were similar in the two groups. CONCLUSION: Atypical CD is not uncommon in India. Children with atypical CD present at an older age. Likelihood of finding CD is high in children with anemia, short stature and rickets. PMID- 18431010 TI - Incidence of hepatocellular carcinoma among Indian patients with cirrhosis of liver: an experience from a tertiary care center in northern India. AB - BACKGROUND/AIM: Despite bearing the main burden of HCC, prospective studies from developing countries are lacking. This prospective observational study was designed to estimate the incidence of HCC among Indian patients with hepatic cirrhosis. METHODS: Between April 2001 and November 2004, we enrolled 301 patients with liver cirrhosis. Patients found to be free of HCC using baseline abdominal ultrasound, triple-phase computed tomography (TPCT) and serum alpha fetoprotein (AFP) levels were followed up prospectively for detection of HCC using ultrasound and AFP every 6 months, and TPCT annually. RESULTS: Among the 194 patients (mean age [SD] 45.1 [+/-13.1] years; male:female 6.1:1.0) followed up, 154 had Child's A and 40 had Child's B disease. The causes of cirrhosis were: hepatitis B-71 (36.6%), hepatitis C-54 (27.8%), dual infection with hepatitis B and C-12 (6.2%) and others including autoimmune, alcoholic and cryptogenic cirrhosis 57 (29.4%). During a cumulative follow up period of 563.4 person-years, 9 cases of HCC were detected, with an incidence rate of 1.60 per 100 person years. CONCLUSION: In our study, the incidence of HCC among patients with liver cirrhosis was intermediate, being lower than that in Japan but higher than that reported from Europe. PMID- 18431011 TI - Clinicopathological predictors to predict sustained viral response rates in patients with chronic hepatitis C infection. AB - BACKGROUND: Treatment of patients with chronic hepatitis C virus (HCV) infection remains suboptimal, with the current pegylated interferon (PEG-IFN) and ribavirin combination therapy providing sustained viral response (SVR) rates of 54 - 63%. The aim of this study was to identify clinical, laboratory and histological findings that can predict non-response to this treatment. METHODS: Medical records of patients who had completed PEG-IFN and ribavirin therapy for chronic HCV infection between December 2002 and November 2005 and had undergone a liver biopsy prior to starting treatment were retrospectively reviewed. Data on various clinical and biochemical parameters were extracted and liver biopsy slides were reviewed by a pathologist who was blinded to the clinical and laboratory findings. RESULTS: Of 67 patients studied (mean [SD] age 46.3 [6.3] years; 36 men), 42/57 (74%) had an early viral response (EVR) and 37/64 (58%) had an SVR. On univariate analysis, absence of EVR (p=0.0002), non-white race (p=0.008), AST/ALT ratio > or = 1.0 (p=0.008), INR > or = 1.0 (p=0.02) and presence of steatosis > or = 5% on liver biopsy (p=0.03) were associated with lack of SVR. In multivariate analysis, all of these except INR were significant independent predictors of SVR. CONCLUSIONS: Absence of EVR, non-white race, AST/ALT ratio > or = 1.0 and presence of steatosis > or = 5% on liver biopsy are independent predictors of absence of SVR in patients with chronic HCV infection receiving PEG IFN and ribavirin combination treatment. PMID- 18431012 TI - Portal venous thrombosis after umbilical vein catheterization. AB - PURPOSE: Portal vein thrombosis has been associated with umbilical venous catheterization. This prospective study was done to determine the incidence of neonatal portal venous thrombosis associated with catheterization of the umbilical vein . METHODS: Neonates who had undergone umbilical vein catheterization for exchange transfusion between March 2003 and March 2004 in Children's Hospital of Tabriz, Iran, were included. Doppler ultrasonography was performed within 1-2 weeks after the removal of the catheter. In the cases with portal venous thrombosis, subsequent serial ultrasonography was performed at intervals of every 1-2 months until clot resolution. Risk factors, if any were identified and correlated with catheter-related thrombi. RESULTS: Ultrasonography detected clinically silent portal venous thrombosis in 17 (34%) of 50 neonates. Follow-up ultrasonography was available in 13 of 17 babies, and revealed complete or partial resolution in all the cases. Sepsis was identified as a significant risk factor (p < 0.001). CONCLUSION: Umbilical venous catheter-associated thrombosis is common, and spontaneous resolution occurs in most cases. PMID- 18431013 TI - Epidemiology of inflammatory bowel disease in Asia. AB - Inflammatory bowel disease (IBD) once considered to be a disease only of the Western world is no longer so. Though good epidemiological studies are available from the West, still the information available from Asia is inadequate. This article is a review of the available epidemiological studies from Asia. The incidence / prevalence rates of IBD are still the highest in Europe and North America, though there is convincing evidence of rising trend of IBD in Asian countries; India probably heads the list. There is a need to study the role of various environmental and genetic factors in different countries in Asia. At present, ulcerative colitis predominates but Crohn's disease is expected to manifest more in coming years. PMID- 18431014 TI - Extensive gastrointestinal tract and thyroid involvement with Wegeners granulomatosis. AB - Wegeners granulomatosis (WG) is a pauci-immune systemic vasculitis involving small to medium sized blood vessels of the respiratory tract and renal vasculature. We report a 34-year-old lady with extensive gastrointestinal tract, pancreas and thyroid involvement. Literature review revealed only two prior reports of esophageal involvement, two reports of pancreatic involvement and few cases of thyroid involvement. PMID- 18431015 TI - Acral and palmo-plantar hyperpigmentation in a patient with disseminated hepatocellular carcinoma. AB - Capecitabine (5-fluorouracil prodrug) is being evaluated for the management of hepatocellular carcinoma, and is associated with a peculiar skin reaction called hand and foot syndrome (HFS). We describe one patient with HCC and drug-induced HFS. PMID- 18431016 TI - Primary intestinal lymphangiectasia as a component of autoimmune polyglandular syndrome type I: a report of 2 cases. AB - Chronic diarrhea and steatorrhea occur frequently in patients with autoimmune polyglandular syndrome (APS) type I. Intestinal lymphangiectasia has been reported earlier as a cause of steatorrhea in a young girl with APS Type I. We describe 2 patients with APS Type I who were found to have intestinal lymphangiectasia, one of whom had symptomatic protein-losing enteropathy. PMID- 18431017 TI - Iron deficiency anemia in Asians and Caucasians -- Any differences? PMID- 18431018 TI - Screening for hepatocellular carcinoma in hepatitis B and C chronic carriers in Iran. PMID- 18431019 TI - HBeAg negative chronic hepatitis B with persistently normal serum transaminase and low HBV DNA can cause significant liver disease. PMID- 18431020 TI - Octreotide in congenital chylous ascites an avoid requirement of total parenteral nutrition. PMID- 18431021 TI - Stomach cancer incidence among males in Golestan province, Iran. PMID- 18431022 TI - Mantle cell lymphoma (multiple lymphomatous polyposis) of gastrointestinal tract. PMID- 18431023 TI - Transient neurotoxicity due to 5-fluorouracil. PMID- 18431024 TI - Portal hypertension associated with sickle cell disease. Is there a coexistent liver disease? PMID- 18431025 TI - Detection of gall bladder cancer metastases in rare sites by PET scan. PMID- 18431026 TI - Mediastinal masquerade. PMID- 18431027 TI - Soluble EMMPRIN (extra-cellular matrix metalloproteinase inducer) stimulates the migration of HEp-2 human laryngeal carcinoma cells, accompanied by increased MMP 2 production in fibroblasts. AB - The basement membrane functions as a barrier against the invasion of cancer cells. It is therefore important to investigate the mechanism of basement membrane degradation by matrix metalloproteinases (MMPs). Previously, cancer cells were long considered to be the major source of MMPs; however, current evidence indicates that most MMPs in cancer tissue are produced by stromal rather than cancer cells. A glycoprotein highly expressed on the cancer-cell membrane, EMMPRIN (extra-cellular matrix metalloproteinase inducer), exhibits the potential role of the MMP inductor in stromal cells. Depending on the cell type, EMMPRIN can stimulate the production of MMP-1, MMP-2, and MMP-3. We here report that soluble full-length EMMPRIN is liberated from HEp-2 human laryngeal epidermoid carcinoma cells, probably via microvesicle shedding. Soluble EMMPRIN stimulates human fibroblasts to produce MMP-2, after which the augmented migration of HEp-2 cells occurs, as observed in an invasion chamber assay with separately cultured fibroblasts. An anti-EMMPRIN function-blocking antibody reduced MMP-2 activity in the conditioned medium and inhibited the migration of HEp-2; obviously, EMMPRIN activity contributes to cancer-cell migration. We postulate that soluble EMMPRIN probably triggers the promotion of cancer invasion in vivo. PMID- 18431028 TI - Reduced expression of endogenous secretory receptor for advanced glycation endproducts in hippocampal neurons of Alzheimer's disease brains. AB - The receptor for advanced glycation endproducts (RAGE) is a cell-surface multiligand receptor, which interacts with amyloid beta (Abeta), a key protein in Alzheimer's disease (AD). RAGE-Abeta interaction is thought to be associated with pathological progression in AD. A splice variant of RAGE, endogenous secretory RAGE (esRAGE) can act as a decoy receptor for RAGE ligands that would prevent the progression of some pathologic conditions. In this study, the expression of esRAGE in the hippocampal tissues from AD brains compared with control (non-AD) was examined by immunohistochemistry and Western blot analysis. Semiquantitative immunohistochemical analysis of hippocampal tissues using esRAGE-specific antibody revealed significantly decreased immunoreactivities in pyramidal cells in CA1 and CA3 regions of AD compared with non-AD. On the other hand, immunoreactivities of astrocytes for esRAGE significantly increased in those regions. Dentate granule cells and astrocytes showed essentially invariant immunoreactivities between AD and non-AD. Changes in esRAGE immunoreactivity in CA3 neurons and astrocytes were observed from the early pathological stages. Moreover, the esRAGE-immunoreactive bands of AD samples were weaker than those of non-AD samples in Western blot analysis. The results indicate that low expression of esRAGE in the hippocampus would be associated with the development of AD. PMID- 18431029 TI - Existence of subtypes of gustducin-immunoreactive cells in the vallate taste bud of guinea pigs. AB - Vallate taste buds in the guinea-pig tongue were immunohistochemically investigated with regard to the colocalization of gustducin with calbindin-D28K (=spot 35 protein) and type III inositol triphosphate receptor (IP(3)R-3) in order to characterize gustducin-immunoreactive cells. Individual taste bud cells ranged from totally immunopositive to totally immunonegative for these three molecules. Among the immunoreactive cells, gustducin-immunoreactive cells were divided into two cell populations: one immunopositive and the other immunonegative for calbindin-D28K. Applying our previous data to the present results, the former cells should belong to Type III cells designated by electron microscopy. This finding provides new evidence regarding the taste bud types of cells expressing gustducin in the guinea pig. PMID- 18431030 TI - Anterograde labeling of the corticospinal tract in jimpy mutant mice by Di I injection into the motor cortex. AB - Carbocyanine fluorescent dye, DiI, is an excellent anterograde/retrograde neural tracer, but its efficacy for the anterograde labeling of neural circuits in the adult brain tends to decrease with ages. The present study shows that an injection of DiI into the motor cortex of the young adult jimpy mutant mice (Plp1(jp)/+) resulted in successful anterograde labeling of corticospinal tract fibers. Furthermore, an injection of Fast Blue into the lumbar spinal cord of the mutant mice resulted in retrograde labeling of layer 5 corticospinal tract neurons within the motor cortex. Since no abnormality except for myelin deficiency is known in the long descending and ascending tracts of jimpy mutant mouse, this mutant is suitable for neural tracing studies of long axonal trajectories with the use of carbocyanine dye, DiI, although these males die between 20 and 40 days of age. PMID- 18431031 TI - Specific depletion of GGA2 causes cathepsin D missorting in HeLa cells. AB - Three mammalian GGAs (Golgi-localized, gamma-ear-containing, ARF-binding proteins), GGA1, 2, and 3 have been implicated in the sorting of mannose 6 phosphate receptor (MPR). To investigate the distinct roles of GGA2 in lysosomal enzyme transport, we established two stable cell lines that had a reduced expression of GGA2 by RNA interference. The expression levels of GGA2 were approximately 5% of the control levels, whereas those of non-targeted GGA1 and GGA3 were not apparently reduced. The depletion of GGA2 did not cause changes in the overall distribution of GGA1, GGA3, cation-dependent MPR, or cation independent MPR. However, the cell lines showed increased secretion of a lysosomal enzyme, cathepsin D. In addition, a moderate expression of the dominant negative VHS-GAT domain of GGA2 had no effect on the trans-Golgi network (TGN) signal of three GGAs, nor was the GGA2 signal affected by the expression of VHS GAT domain of GGA1 or 3. These results suggest that GGA2 is recruited to the TGN independently of the other GGAs and is required for the efficient sorting of lysosomal enzymes. PMID- 18431032 TI - The differential distribution of type IV collagen alpha chains in the subepithelial basement membrane of the human alimentary canal. AB - We studied distribution patterns of type IV collagen alpha chains in the subepithelial basement membrane (SBM) of the human gastrointestinal tract - the esophagus through the anal canal - by immunofluorescent microscopy using alpha(IV) chain-specific monoclonal antibodies. The alpha1(IV), alpha2(IV), alpha5(IV), and alpha6(IV) chains were found in the SBM throughout the tract, indicating the localization of [alpha1(IV)](2)alpha2(IV) and [alpha5(IV)](2)alpha6(IV) heterotrimeric molecules. The [alpha1(IV)](2)alpha2(IV) molecule was continuously stained, while the [alpha5(IV)](2)alpha6(IV) molecule was weakly stained in gastric glands and small intestinal crypts. In addition, the SBM at the luminal surface epithelium of the stomach and large intestine contained small amounts of alpha3(IV) and alpha4(IV) chains which combined to form the alpha3(IV)alpha4(IV)alpha5(IV) heterotrimeric molecule with alpha5(IV) chain. The SBM beneath the villous epithelium of the small intestine was also demonstrated to have an alpha3(IV) chain and alpha4(IV) chain. Considering the specific locations of the type IV collagen trimers throughout the gastrointestinal SBM, the supramolecular network containing the alpha3(IV)alpha4(IV)alpha5(IV) molecule appears to function as a selective permeability barrier and /or as a protection against chemical stress from the luminal digestive enzymes. PMID- 18431033 TI - Relationships of job hazards, lack of knowledge, alcohol use, health status and risk taking behavior to work injury of coal miners: a case-control study in India. AB - Objective is to assess the relationships of job hazards, individual characteristics, and risk taking behavior to occupational injuries of coal miners. This case-control study compared 245 male underground coal miners with injury during the previous two-year period with 330 matched controls without injury during the previous five years. Data were collected via face-to-face interview and analyzed using the conditional logistic model. Handling material, poor environmental/working conditions, and geological/strata control- related hazards were the main risk factors: adjusted ORs 5.15 (95% CI 2.42-10.9), 2.40 (95% CI 1.29-4.47), and 2.25 (95% CI 1.24-4.07) respectively. Their roles were higher among the face-workers than among the non-face-workers. No formal education, alcohol consumption, disease, big-family, and risk-taking behavior were associated with injuries (2.360.1 ng/ml) showed significantly high plasma levels of calcium, CML and pentosidine as compared with the cTnT-negative group. In multiple logistic regression analysis, the prevalence of patients with high plasma calcium (>median) was increased in the cTnT-positive group as compared with the cTnT negative group (OR: 5.08, 95% CI: 1.62-15.92, p < 0.01). The prevalence of high plasma CML (>median) was increased in the cTnT-positive group (OR: 4.45, 95% CI: 1.41-14.03, p < 0.05). Further, the prevalence of high plasma pentosidine (>median) was also increased in the cTnT-positive group (OR: 4.94, 95% CI: 1.55 15.70, p < 0.01). CONCLUSION: In addition to calcium, AGEs such as CML and pentosidine were associated with cTnT, a marker of myocardial damage, in HD patients. PMID- 18431052 TI - Developmental and regional patterns of GAP-43 immunoreactivity in a metamorphosing brain. AB - Growth-associated protein-43 is typically expressed at high levels in the nervous system during development. In adult animals, its expression is lower, but still observable in brain areas showing structural or functional plasticity. We examined patterns of GAP-43 immunoreactivity in the brain of the bullfrog, an animal whose nervous system undergoes considerable reorganization across metamorphic development and retains a strong capacity for plasticity in adulthood. Immunolabeling was mostly diffuse in hatchling tadpoles, but became progressively more discrete as larval development proceeded. In many brain areas, intensity of immunolabel peaked at metamorphic climax, the time of final transition from aquatic to semi-terrestrial life. Changes in intensity of GAP-43 expression in the medial vestibular nucleus, superior olivary nucleus, and torus semicircularis appeared correlated with stage-dependent functional changes in processing auditory stimuli. Immunolabeling in the Purkinje cell layer of the cerebellum and in the cerebellar nucleus was detectable at most developmental time points. Heavy immunolabel was present from early larval stages through the end of climax in the thalamus (ventromedial, anterior, posterior, central nuclei). Immunolabel in the tadpole telencephalon was observed around the lateral ventricles, and in the medial septum and ventral striatum. In postmetamorphic animals, immunoreactivity was confined mainly to the ventricular zones and immediately adjacent cell layers. GAP-43 expression was present in olfactory, auditory and optic cranial nerves throughout larval and postmetamorphic life. The continued expression of GAP-43 in brain nuclei and in cranial nerves throughout development and into adulthood reflects the high regenerative potential of the bullfrog's central nervous system. PMID- 18431054 TI - Development of the vestibular apparatus and central vestibular connections in a wallaby (Macropus eugenii). AB - We have studied the early development of the vestibular apparatus and its central connections in the tammar wallaby (Macropus eugenii) in order to determine whether the vestibular system anatomy is sufficiently mature at birth to assist in climbing to the pouch. Structural development was studied with the aid of hematoxylin and eosin stained sections and immunoreactivity for GAP-43, whereas the development of vestibular system connections was examined by carbocyanine dye tracing. At the time of birth, the otocyst has distinct utricle, saccule and semicircular canals with immature sensory regions receiving innervation by GAP-43 immunoreactive fibers. Vestibular nerve fibers can be traced into the brainstem to the developing vestibular nuclei, which are not yet cytoarchitectonically distinct. The vestibular nuclei do not contribute direct projections to the lower cervical spinal cord at birth; most bulbospinal projections in the newborn appear to be derived bilaterally from the gigantocellular, lateral paragigantocellular reticular and ventral medullary nuclei. A substantial bilateral projection to the vestibular ganglion and apparatus from the region of the gigantocellular and lateral paragigantocellular nuclei was seen at birth, but not in subsequent ages. This is similar to a projection seen in newborn Ameridelphians. By postnatal day (P) 5, the vestibular apparatus had extensive projections to all vestibular nuclei and neurons projecting in the lateral vestibulospinal tract could be identified in the lateral vestibular nucleus. Cytoarchitectonic differentiation of the vestibular nuclei proceeded over the next 3 to 4 weeks with the emergence of discrete parvicellular and magnocellular components of the medial vestibular nucleus by P19. GAP-43 immunoreactivity stayed high in the lateral vestibulospinal tract for several months after birth, suggesting that the development of this tract followed a prolonged timecourse. Our findings indicate that central and peripheral connections of the vestibular ganglion are present at birth, but that there is no direct projection from the vestibular nuclei to the cervical spinal cord until P5. Nevertheless, the possibility remains that an indirect projection between the vestibular nuclei and the medial reticular formation is present at birth and mediates control of the climb. PMID- 18431053 TI - Nest of origin predicts adult neuron addition rates in the vocal control system of the zebra finch. AB - Neurogenesis and neuronal replacement in adulthood represent dramatic forms of plasticity that might serve as a substrate for behavioral flexibility. In songbirds, neurons are continually replaced in HVC (used as a proper name), a pre motor region necessary for the production of learned vocalizations. There are large individual differences in HVC neuron addition. Some of this variation is probably due to individual differences in adult experience; however, it is also possible that heritability or experience early in development constrains the levels of adult neuron addition. As a step toward addressing the latter two possibilities, we explored the extent to which nest of origin predicts rates of HVC neuron addition in adult male zebra finches. One month after injections of [(3)H]-thymidine to mark dividing cells, neuron addition in HVC was found to co vary among birds that had been nest mates, even when they were housed in different cages as adults. We also tested whether nest mate co-variation might be due to shared adult auditory experience by measuring neuron addition in nest mate pairs after one member was deafened. There were significant differences in neuron addition between hearing and deaf birds but nest mate relationships persisted. These results suggest that variation in genotype and/or early pre- or postnatal experience can account for a large fraction of adult variation in rates of neuron addition. These results also suggest that a major constraint on neurogenesis and the capacity to adjust rates of neuron addition in response to adult auditory experience is established early in development. PMID- 18431055 TI - Brain organization and specialization in deep-sea chondrichthyans. AB - Chondrichthyans occupy a basal place in vertebrate evolution and offer a relatively unexplored opportunity to study the evolution of vertebrate brains. This study examines the brain morphology of 22 species of deep-sea sharks and holocephalans, in relation to both phylogeny and ecology. Both relative brain size (expressed as residuals) and the relative development of the five major brain areas (telencephalon, diencephalon, mesencephalon, cerebellum, and medulla) were assessed. The cerebellar-like structures, which receive projections from the electroreceptive and lateral line organs, were also examined as a discrete part of the medulla. Although the species examined spanned three major chondrichthyan groupings (Squalomorphii, Galeomorphii, Holocephali), brain size and the relative development of the major brain areas did not track phylogenetic groupings. Rather, a hierarchical cluster analysis performed on the deep-sea sharks and holocephalans shows that these species all share the common characteristics of a relatively reduced telencephalon and smooth cerebellar corpus, as well as extreme relative enlargement of the medulla, specifically the cerebellar-like lobes. Although this study was not a functional analysis, it provides evidence that brain variation in deep-sea chondichthyans shows adaptive patterns in addition to underlying phylogenetic patterns, and that particular brain patterns might be interpreted as 'cerebrotypes'. PMID- 18431056 TI - Malabsorption syndrome. Introduction. PMID- 18431057 TI - Intestine: organ or apparatus? AB - It is well known that human intestine is involved in different important functions. First of all, it is responsible for digestion and absorption of nutrients, electrolytes, water, bile salts and drugs, but it also has immunologic, endocrine and motor functions. Moreover, intestinal microflora, composed by a large diversity of bacterial cells, provides several beneficial functions for the host and is, nowadays, defined by many authors as an organ itself. In consideration of intestine complexity, we tried to understand if it can be considered only an organ or if it is an apparatus itself. We have analyzed the different components and their relationships, showing that a continuous collaboration is required among enterocytes, endocrine intestinal cells, gut immune system and microflora to assure an efficient mechanism of defense. In consideration of the complexity of intestinal components, together with the emergent role of microflora, we think that we could start to consider gut as a real apparatus, and not only as an organ. PMID- 18431058 TI - Adverse reactions to food: allergies and intolerances. AB - All the anomalous reactions secondary to food ingestion are defined as 'adverse reactions to food'. In 1995 the European Academy of Allergology and Clinical Immunology suggested a classification on the basis of the responsible pathogenetic mechanism; according to this classification, non-toxic reactions can be divided into 'food allergies' when they recognize immunological mechanisms, and 'food intolerances' when there are no immunological implications. The diagnostic approach to adverse reactions to food is based on accurate clinical history and objective examination, and further execution of specific tests when allergy or intolerance is suspected. The therapy for food allergies is the elimination of the food to which hypersensibility has been found; this strategy can lead, especially in pediatric age, to tolerance. If elimination diets cannot be completely performed, or if it is not possible to identify the food to eliminate, some drugs (e.g. antihistaminics, steroids, etc.) can be administered. Specific allergen immunotherapy has been recently introduced. Fundamental is food allergy prevention, especially in high-risk subjects. The therapeutic approach to secondary food intolerances is based principally on primitive disease resolution; on the other hand, some specific treatments (e.g. beta-galactosidases in lactose malabsorption) are available in case of primary intolerance. PMID- 18431059 TI - Classification of malabsorption syndromes. AB - Malabsorption syndrome is usually defined as the complex of symptoms secondary to maldigestion and/or malabsorption, realizing when the extension of the disease exceeds the ability of intestine compensation. Several conditions have been recognized as being responsible for this syndrome. Up to now, different criteria have been used to order them, but a definitive classification is still not available because of the complexity of the absorption process, the involvement of different organs and structures, and the coexistence of different mechanisms in some diseases causing malabsorption. We propose a new classification of diseases causing malabsorption syndrome according to the responsible etiopathogenetic mechanisms: (a) alteration of digestive processes; (b) alteration of uptake and transport caused by damage or reduction of absorption surface, and (c) miscellaneous. A comment about the mechanisms responsible for malabsorption is given for all the cited diseases. PMID- 18431060 TI - A history of coeliac disease. AB - Coeliac disease may have an ancient history dating back to the 1st and 2nd centuries AD. The first clear description was given by Samuel Gee in 1888. He suggested that dietary treatment might be of benefit. In the early 20th century various diets were tried, with some success, but without clear recognition of the toxic components. The doctoral thesis of Wim Dicke of 1950 established that exclusion of wheat, rye and oats from the diet led to dramatic improvement. The toxicity was shown to be a protein component, referred to as gluten. Dicke's colleagues, Weijers and Van de Kamer, showed that measurement of stool fat reflected the clinical condition. Early studies were in children but stool fat measurements documented that the condition could be recognised in adults. Histological abnormalities of the lining of the small intestine were demonstrated beyond doubt by Paulley in 1954 and techniques of per-oral biopsy described by Royer in 1955 and Shiner in 1956 afforded reliable diagnosis. Concurrence in monozygotic twins suggested a genetic component, confirmed by studies of HLA antigens. Additional, non-genetic factors seem likely. Circulating antibodies suggest an immunological mechanism of damage and provide non-invasive screening tests. Lymphoma, adenocarcinoma and ulceration of the small intestine and a range of immunological disorders are associated. A relationship with dermatitis herpetiformis was suggested by Samman in 1955 and established by Shuster and Marks in 1965 and 1968. The Coeliac Society (now Coeliac UK) was founded in 1968 and similar societies now exist across the world. They provide an extremely valuable service. Present problems include definition of the tolerated levels of gluten, whether oats are toxic for some or all coeliacs and the likelihood that the condition is relatively common and frequently without classical symptoms. Hope for the future is that more convenient methods of treatment will follow better understanding. PMID- 18431061 TI - Celiac disease: what's new about it? AB - In the present review we will try to summarize the clinical and diagnostic features of celiac disease (CD) as well as the new findings on extraintestinal manifestation. CD is an immune-mediated enteropathy caused by a permanent gluten intolerance. In the last years, the diagnosis is becoming more and more frequent because of the recognition of 'new' symptoms and associated extraintestinal manifestations. Classical CD is dominated by symptoms and sequelae of gastrointestinal malabsorption. In the 'atypical forms', the extraintestinal features usually predominate, with few or no gastrointestinal symptoms. Silent CD refers to asymptomatic patients with a positive serologic test and villous atrophy on biopsy. This form is detected by screening of high-risk individuals, or villous atrophy occasionally may be detected by endoscopy and biopsy conducted for another reason. The potential form is diagnosed in groups at risk including relatives of celiac patients, Down syndrome and autoimmune diseases. Latent CD is defined by positive serological tests but not histological changes on biopsy. These individuals are asymptomatic, but later may develop symptoms and/or histological alterations. Recognition of atypical manifestations of CD is very important because many cases can remain undiagnosed with an increased risk of long-term complications. PMID- 18431062 TI - Metabolic and nutritional features in adult celiac patients. AB - Celiac disease (CD) is a chronic immune-mediated gluten-dependent enteropathy induced by ingestion of gluten-containing products, characterized by intestinal malabsorption and subtotal or total atrophy of intestinal villi, which improves after gluten-free diet (GFD). Untreated patients affected by the classic form of CD are at high risk of malnutrition, but an impairment of nutritional status is frequently reported also in patients with the subclinical form of the disease. Strict adherence to a GFD greatly improves nutritional status, inducing an increase in fat and bone compartments, but does not completely normalize body composition. A lack of improvement in nutritional status may identify incomplete adherence to GFD treatment. Evidence has shown lower body weights and lower fat mass and fat-free mass contents in CD patients. Untreated CD patients oxidize more carbohydrates as energy substrate compared to treated subjects. In addition, circulating ghrelin concentration was reduced after GFD treatment as a possible consequence of body composition improvement, while leptin did not correlate with the changes in body composition and substrate oxidation in patients with CD. A significant correlation was reported between ghrelin and the degree of severity of intestinal mucosal lesions. CD patients might show an alteration in lipid metabolism, i.e. low serum total and high- density lipoprotein-cholesterol as a consequence of lipid malabsorption and decreased intake. In conclusion, weight loss and nutritional deficiencies are relevant clinical features in CD. Thus, an early and accurate evaluation of nutritional status and energy metabolism represents a fundamental tool in the management of CD patients. PMID- 18431063 TI - Utility of double-balloon enteroscopy for the evaluation of malabsorption. AB - INTRODUCTION: Occasionally, patients with malabsorption represent a diagnostic challenge. Double-balloon enteroscopy (DBE) allows deep and detailed examination of the small bowel. AIM: To determine the diagnostic value of DBE in patients with malabsorption of unclear origin. METHODS: DBE was performed in a total of 12 patients with clinical malabsorption. Biopsy specimens were taken from macroscopic lesions or from examined small bowel at three different levels of scope insertion depth. Tissue specimens were evaluated with standard hematoxylin and eosin, the modified Marsh classification and, when indicated, special stains for amyloidosis. RESULTS: Fifteen DBEs were successfully performed in 12 patients without complications. DBE with small bowel biopsies yielded a diagnosis in 8 patients (67%). A new diagnosis was reached in 4 patients (33%). The new diagnoses included: Crohn's disease, primary intestinal lymphangiectasia and jejunal amyloidosis. In none of these 4 patients did the duodenal biopsies yield a diagnosis. Also, DBE excluded enteropathy-associated T-cell lymphoma (EATL) and/or ulcerative jejunitis in symptomatic celiac disease patients. CONCLUSIONS: DBE had a diagnostic value of 42% in patients with malabsorption of unclear origin. In addition, DBE was useful to rule out complications of long-standing celiac disease such as ulcerative jejunitis or EATL. DBE should be reserved for patients with unexplained malabsorption. DBE with jejunal and ileal biopsies appears to have a diagnostic value in patients with malabsorption, even when duodenal biopsies are histologically normal. PMID- 18431064 TI - Affective and psychiatric disorders in celiac disease. AB - Several extraintestinal clinical manifestations have been reported in celiac disease (CD). Among them, growing evidence suggests the association between CD and affective and psychiatric disorders. In this review the most frequent affective and psychiatric disorders associated with CD and the possible mechanisms involved in these associations were analyzed. The available data suggest that screening for CD in patients with affective and/or psychiatric symptoms may be useful since these disorders could be the expression of an organic disease rather than primary psychiatric illnesses. PMID- 18431065 TI - Vascular involvement in inflammatory bowel disease: pathogenesis and clinical aspects. AB - Crohn's disease (CD) and ulcerative colitis (UC), the two major forms of inflammatory bowel disease (IBD), are chronic inflammatory conditions, characterized by a microvascular and also macrovascular involvement. Chronically inflamed intestinal microvessels of IBD patients have demonstrated significant alterations in their physiology and function compared with vessels from healthy and uninvolved IBD intestine. Recently, some studies have revealed that the poor mucosal healing, refractory inflammatory ulcerations and damage in the IBD intestine could depend on microvascular dysfunction, resulting in diminished vasodilatory capacity and tissue hypoperfusion in the IBD gut. Furthermore, several data show that the activation of intestinal endothelium plays a critical role in the pathogenesis and/or in perpetuating and amplifying the inflammatory process in IBD and, consequently, it is now emerging as a potential use of anticoagulant or coagulation-related drugs in treating IBD. IBD is also associated with an increased risk of macrovascular venous and arterial thrombosis. Thrombotic events occur prevalently as deep vein thrombosis and pulmonary embolism. They happen at an earlier age than in non-IBD patients. Prothrombotic risk factors in IBD patients could be distinguished as acquired, such as active inflammation, immobility, surgery, steroid therapy, and use of central venous catheters, and inherited. Furthermore, it has been found that IBD, per se, is an independent risk factor for thrombosis. The prevention of thromboembolic events in IBD patients includes the elimination of removable risk factors and, if thrombosis occurs, a pharmacological therapy similar to that used for thromboembolic events occurring in the general population. PMID- 18431066 TI - Role of the brain-gut axis in the pathophysiology of Crohn's disease. AB - Studies on the interaction between the central nervous system and the gastrointestinal system have shed light on the neurobiological response to stress via the hypothalamic-pituitary-adrenal and the hypothalamic-autonomic nervous system axes. These findings support a role of psychological and environmental factors in the course of gastrointestinal disorders and their influence on the neuroendocrine regulation of the immune system. Crohn's disease (CD) is a chronic inflammatory condition of the gastrointestinal tract, whose etiology involves genetic, psychological, immune and inflammatory factors. A higher prevalence of psychiatric diagnosis has been observed in CD patients. Both longitudinal and cross-sectional studies have explored the relationship between psychological stress and severity and/or clinical course of CD, with different, even conflicting, results. In several chronic diseases and stress-related psychological disorders, an alteration has been observed of the HPA response that through glucocorticoids modulate the immune/inflammatory reaction. In animal models of colitis, psychological or environmental stress may increase gastrointestinal permeability, allowing abnormal antigen presentation to the immune system and leading to the exacerbation and perpetuation of intestinal inflammation. The increased intestinal permeability under stress is mediated by corticotropin-releasing hormone stimulation through nicotinic, adrenergic and cholinergic receptors, suggesting a complex interplay between sympathetic and parasympathetic nervous systems. This review will examine and discuss the relationship between psychological stress and CD, investigating the role played by the hypothalamic-pituitary-adrenal and the hypothalamic-autonomic nervous system axes in stress-related psychological disorders, and the possible influence of chronic stress on the intestinal inflammation, in particular in CD. PMID- 18431067 TI - Intestinal malabsorption and skin diseases. AB - Several skin manifestations were described in patients affected by intestinal disorders. The development of skin diseases in these patients could be related to the impairment of intestinal absorption and motility, other than to immunological and hormonal changes. The growing evidence of the association between skin disorders and intestinal diseases suggests that the skin could be considered the 'mirror of the gut'. PMID- 18431068 TI - Probiotics: which and when? AB - There is a natural feeling between our intestinal flora and the gut. These microorganisms, living in the various tracts of human intestine, may affect the host homeostasis. Some of these bacteria can perhaps be a source of infection and sepsis when the bowel barrier is physically or functionally breached. The term 'probiotic' dates from the beginning of the last century and in the last years a market for probiotics worldwide, estimated to be worth billions of pounds, has developed. Although there is persuasive advertising for probiotics and there have been methodological advances in the study of the intestinal microbiota, much remains unproven, e.g. how probiotics work, which strains are effective, what can be expected to be achieved, and what dosage is required for effectiveness. This review of the literature is an evidence-based guide through the developing microbial universe affecting our life. PMID- 18431069 TI - Fecal calprotectin concentrations in patients with small intestinal bacterial overgrowth. AB - BACKGROUND/AIMS: Small intestinal bacterial overgrowth (SIBO) is defined by any condition in which the proximal part of the small bowel harbors for a long time > 10(5) bacteria/ml of the intestinal juice. No data are currently available about direct or indirect parameters indicating the presence of leukocytes in the gut wall and mucosal neutrophil turnover in patients with SIBO. In our pilot study we evaluate fecal calprotectin concentrations (FCC) in patients with SIBO in order to identify a possible presence of subclinical intestinal inflammation. METHODS: 40 consecutive patients with SIBO resulting positive to hydrogen glucose breath test, and 40 adult healthy volunteers were included in the study. FCC were determined by ELISA. Mean FCC were compared by means of the t-test for independent samples. RESULTS: FCC in patients with SIBO were not significantly different compared to controls (p = 0.907). CONCLUSION: Our study shows for the first time that FCC in patients with SIBO do not significantly differ from controls, suggesting that in SIBO there are no intestinal subclinical inflammatory changes involving principally the neutrophils. PMID- 18431070 TI - Antiphospholipid syndrome manifested by gastrointestinal bleeding: can we overlook endoscopically revealed small lesions? PMID- 18431071 TI - Lipoleiomyoma of the uterus: imaging features. AB - BACKGROUND: Lipoleiomyoma is an extremely rare, benign, uterine tumor that requires no treatment when asymptomatic. It is important to differentiate it from malignant conditions requiring surgery. METHODS: A uterine lipoleiomyoma in a 68 year-old asymptomatic woman exhibited highly suggestive features by ultrasound, multislice spiral computed tomography (CT), and magnetic resonance imaging (MRI). RESULTS: The ultrasound findings were non-specific. CT scan showed an 8 x 7 cm well-defined fatty mass of the uterine body. On T(2)-weighted MR images, fat content within the tumor was confirmed because of evident chemical shift artifact.To our knowledge, the findings provided by these three modalities used in combination have not been reported previously in the gynecologic literature. CONCLUSION: The data yielded by CT and MRI on the fatty nature of the lesion are valuable in diagnosing this entity.The final pathological examination results confirmed the diagnosis of lipoleiomyoma. PMID- 18431072 TI - Variants of parvovirus B19: bioinformatical evaluation of nested PCR assays. AB - Variants of parvovirus B19 are currently grouped into three genotypes: 1 (reference B19 strains), 2 and 3. It has been evidenced that isolate K71 of genotype 2 is more prevalent in skin than the conventional B19 genotype 1. In this study we investigated the detection of parvovirus B19 genotypes by using two nested PCRs and evaluating the suitability of these assays by BLAST search of parvovirus isolates. Subsequently, we analyze the present genotypes in skin biopsies. The two nested PCRs employed in this study allow to amplify 41 isolates as confirmed by bioinformatical validation. The molecular epidemiological characterization of our casistics confirmed the presence of isolate K71 in human skin. PMID- 18431073 TI - The regulation of mesangial cell proliferation. AB - Mesangial cell (MC) proliferation is a key pathological feature in a number of common human renal diseases including IgA, systemic lupus erythematosus and diabetic nephropathies. Knowledge of the role of MCs in normal glomeruli and of their response to pathological stimuli is crucial to the understanding of these disease processes. The purpose of understanding disease is ultimately to develop therapeutic strategies that can limit or even reverse the underlying pathological process. Over the last 20 years a number of signaling pathways involved in the regulation of MC proliferation have been identified and studied with a view to manipulating them for therapeutic gain. Unfortunately, despite these advances, there are still very few clinical options that specifically target aberrant MC proliferation. This article reviews a number of factors that have been shown to play a role in controlling MC proliferation, including signaling molecules (e.g. Platelet-derived growth factor, Ras and Ca(2+)), cell cycle proteins (e.g. cyclin D1) and transcription factors (E2F). A variety of strategies has been used to manipulate these different pathways to elucidate their function in MCs with the ultimate aim of modifying them in order to treat human renal diseases. PMID- 18431074 TI - Lack of a significant effect of cannabinoids upon the uptake of 2-deoxy-D-glucose by Caco-2 cells. AB - The endogenous cannabinoid system plays a role in the regulation of energy homeostasis acting through central pathways, and its dysregulation may be implicated in the pathogenesis of obesity. Recent evidence is accumulating showing that the endogenous cannabinoid system is also present in peripheral tissues. The aim of this work was to investigate the effect of cannabinoids upon the intestinal absorption of glucose. For this, we investigated the effect of some cannabinoid receptor agonists and antagonists upon the apical uptake of 3H-2 deoxy-D-glucose by the human intestinal epithelial Caco-2 cells. Uptake of a low concentration of 3H-2-deoxy-D-glucose (1 micromol/l) was both cytochalasin B- and phloridzin-sensitive. The maximal inhibition obtained with each of these inhibitors was 50%, and their effect was not cumulative. On the other hand, uptake of a high concentration of 3H-2-deoxy-D-glucose (20 mmol/l) was partially inhibited by cytochalasin B (+/-20%) and phloridzin had no effect. We verified that neither the cannabinoid receptor agonists [tetrahydrocannabinol (1-10 micromol/l), anandamide (0.1-10 micromol/l) and CP 55,940 (5 nmol/l to 1 micromol/l)], nor the specific CB1 and CB2 antagonists [AM251 (10-500 nmol/l) and AM630 (50 nmol/l to 1 micromol/l), respectively] had a significant effect upon 3H 2-deoxy-D-glucose uptake by Caco-2 cells. This was true for both the uptake of a low (1 micromol/l) and of a high (20 mmol/l) concentration of 3H-2-deoxy-D glucose. From these results, we may hypothesize that cannabinoids do not interfere with the intestinal GLUT2-mediated apical uptake of glucose. PMID- 18431076 TI - Upper limb movements and cerebral plasticity in post-stroke rehabilitation. AB - Rehabilitative interventions for the plegic/paretic upper limb of stroke survivors are more effective if they are early, intensive, and provide multisensory stimulation. Various rehabilitative approaches have been proposed to date, but little has been published on clinical efficacy. The mechanism underlying recovery of neurological injury after stroke is still incompletely understood, but more than one process is probably involved and cerebral plasticity undoubtedly plays a key role. The goal of this review was to identify which movements and type of therapeutic arm exercises may influence cerebral plasticity in plegic/paretic stroke survivors. Evidence suggests that plasticity is stimulated more by the arm's movement trajectory than by its final position in space. Rehabilitation should be based on simple, repetitive, unidirectional or, better still, complex and multidirectional movements in all spatial planes, such as circular or spiral movements. It should also incorporate a feedback system, since this seems to bring about earlier and better motor and functional outcomes. PMID- 18431075 TI - Men: good health and high mortality. Sex differences in health and aging. AB - This review examines sex differences in health and survival, with a focus on the Nordic countries. There is a remarkable discrepancy between the health and survival of the sexes: men are physically stronger and have fewer disabilities, but have substantially higher mortality at all ages compared with women: the so called male-female health-survival paradox. A number of proposed explanations for this paradox are rooted in biological, social, and psychological interpretations. It is likely to be due to multiple causes that include fundamental biological differences between the sexes such as genetic factors, immune system responses, hormones, and disease patterns. Behavioral differences such as risk-taking and reluctance to seek and comply with medical treatment may also play a role. Another consideration is that part of the difference may be due to methodological challenges, such as selective non-participation and under-reporting of health problems, and delayed seeking of treatment by men. The Nordic countries provide a unique opportunity for such studies, as they have good-quality data in their national health registers, which cover the whole population, and a long tradition of high participation rates in surveys. PMID- 18431077 TI - Orthogeriatric Unit: a thinking process and a working model. PMID- 18431078 TI - Orthogeriatric care for the elderly with hip fractures: where are we? AB - Hip fracture (HF) is a major health care problem in the Western world, associated with significant morbidity, mortality and loss of function. Its incidence is expected to increase as the population ages. The authors discuss the role of a coordinated multidisciplinary team in the management of patients during hospital stay, at discharge and during rehabilitation. Orthogeriatric care should not just be viewed as a multidisciplinary activity, but as a radical alternative to the traditional model of care, an alternative based on all those strategies in which evidence shows an improvement in outcomes in the fractured elderly. Therefore, key points of the care are early surgery, immediate mobilization, prevention and management of delirium, pain and malnutrition, as well as an integrated and multidisciplinary approach. Comprehensive geriatric assessment is useful in identifying frail elderly and in providing information that is essential in formulating clinical recommendations and making care plans. In each hospital, the orthogeriatric unit should represent a center of excellence for treating elderly patients with major fractures. However, when an orthogeriatric project is implemented, it is essential that detailed data about the case-mix of patients, process of care and outcomes are collected, to compare the results with historical data and to be able to participate in audit processes. PMID- 18431079 TI - Gait variability while dual-tasking: fall predictor in older inpatients? AB - BACKGROUND AND AIMS: Increased gait variability is associated with a high risk of falling in older community-dwellers, but no information exists about the relationship between increased gait variability and falls occurring in older hospitalized patients. We therefore sought to determine, in an acute geriatric setting, whether gait variability in single- (i.e., usual walking) or dual-task conditions can predict inpatient falls. METHODS: Stride time variability was calculated in both single-task (i.e., usual walking) and dual-task conditions with a GAITRite-System in 13 male and 44 female patients (mean age=85.0, SD=6.6 yrs) consecutively admitted to the acute care geriatric department of Geneva University Hospitals, Switzerland. All participants were able to walk without assistive devices at day 3 post-admission. Falls during hospital stay were identified through the hospital accident reporting system. RESULTS: Ten fallers and 47 non-fallers were identified. The first fall events were significantly associated with the coefficient of variation of stride time in both walking conditions during hospital stay (OR 13.3, (95% CI 1.6-113.6), p=0.018 for usual walking; OR 8.6, (95% CI 1.9-39.6), p=0.006 for dual- task walking). Furthermore, the time elapsing between the first day of hospitalization and the first fall was significantly shorter when the cut-off value of stride time variability was calculated for dual-tasking compared with usual walking. The Cox regression model revealed that only the coefficient of variation of stride time during dual-task walking was significantly associated with the occurrence of the first fall event (p=0.006). CONCLUSION: Our results suggest that the degree of stride time variability in dual-task walking conditions distinguished fallers from non fallers in a group of independently walking, older inpatients. PMID- 18431080 TI - Gait reveals bilateral adaptation of motor control in patients with chronic unilateral stroke. AB - BACKGROUND AND AIMS: Functional brain imaging has shown that bilateral brain reorganization may occur after unilateral cerebral damage. The present study searched for evidence of bilateral motor control changes in gait in patients with chronic unilateral stroke. METHODS: Gait variables (temporal and spatial parameters, footprint peak times (FPPT) and footfall times (FFT)) were recorded in 48 patients with chronic unilateral stroke at their preferred speed, and in 10 healthy volunteers walking from very slowly to very fast on a pressure sensor walkway. The data were divided into 4 groups according to gait velocity. The functional outcome of stroke was measured by the Barthel Index. RESULTS: Patients' gait variables reflected their hemiparetic gait pattern. Slower patients had prolonged stance, FPPT and FFT on the non-affected side (NS), and prolonged swing and shorter FPPT and FFT on the affected side (AS). The magnitude of the asymmetry index of these parameters was inversely associated with velocity performance and, at the same time, it characterized how much the control of the clinically healthy (NS) side was altered during walking. Bilateral changes in kinetic and footfall variables in the chronic stage of unilateral stroke were present. CONCLUSIONS: Patients chose their preferred walking velocity using stereotyped, alternative gait patterns, in which the contribution of the NS was larger than that of the AS. Alternative gait patterns may partly demonstrate compensatory behavioral strategies adapted by patients. PMID- 18431081 TI - Ethnic differences in physical performance in older Americans: data from the Third National Health and Nutrition Examination Survey (1988-1994). AB - BACKGROUND AND AIMS: Impaired physical performance in older adults has been shown to vary by ethnic groups. The objective of this study is to examine ethnic differences in physical performance in older Americans. METHODS: We conducted a cross-sectional analysis using a sample of 4456 non-institutionalized Americans aged 60 and above (2712 non- Hispanic Whites, 861 non-Hispanic Blacks and 883 Mexican Americans) from a large national representative survey (The Third National Health and Nutrition Examination Survey (NHANES III) conducted in 1988- 1994). Measurements included socio-demographic variables; self-reported physician diagnosed medical conditions, body mass index (BMI) and physical performance examination (tandem stand balance, timed chair stand, and timed 8-foot walk). RESULTS: Of 4456 study participants, 60.9% (n=2712) subjects were non-Hispanic white, 19.3% (n=861) were non-Hispanic black, and 19.8% (n=883) were Mexican American. About 35% (n=1573) of subjects could not hold the tandem balance test for 10 seconds. Older age, diabetes, stroke and arthritis were significantly associated with decreased performance on the balance test. Older age, female gender, being Mexican American or non-Hispanic black, hip fracture and high BMI were significantly associated with decreased performance on 8- foot walking test, while older age, female gender, being non-Hispanic black, low education, stroke, cancer, arthritis and high BMI were significantly associated with decreased performance on the chair stand test. CONCLUSIONS: With some ethnic variation, older age, female gender, being non-Hispanic black, being Mexican American, low education, high BMI, diabetes, stroke, cancer and arthritis were associated with decreased physical performance in elderly Americans. PMID- 18431082 TI - A 30-year survey of drug use in the 1914 birth cohort in Glostrup County, Denmark: 1964-1994. AB - BACKGROUND AND AIMS: Use of drugs increases with age. Several drugs as well as multiple drug intake are recognized risk factors for symptoms, disability, hospitalizations, and even mortality, due to side effects and problems with compliance. Yet, little is known about the long-term effects. As a first step, this longitudinal study of a general population gives insight into consumption patterns including multiple drug intake ("polypharmacy") and individual adherence to drug use over a 30-year period. METHODS: This is a study based on the first large longitudinal population study of the health of adult Danes. Examinations were performed at 50, 60, 70 and 80 years of age. The Statistical Package for the Social Sciences (SPSS) was used to check participants' individual adherence to each of six main drug categories, calculated stepwise. RESULTS: 802 persons were included at 50 years of age in 1964. Of these, 213 were still participating in the study in 1994. The general use of drugs increased for all drug categories with age. Women used statistically significantly more drugs than men and showed a higher degree of polypharmacy. The individual adherence to antihypertensives was greater than the corresponding adherences to analgesics, psychoactive drugs, or hypnotics. At least two-thirds of all remaining initial users of antihypertensives still took them at 80 years of age. For other drug categories, the corresponding values were half or less. CONCLUSIONS: For the initial population studied over a period of 30 years, the use of drugs increased with age, but the drugs were not prescribed for the same people over time. During the 30-year period, individual patients' drug adherence habits were in accordance with scientific evidence: patients adhered to long-term use of antihypertensive drugs, but used analgesics, psychoactive drugs and hypnotics for only shorter periods. PMID- 18431083 TI - Low plasma selenium concentrations and mortality among older community-dwelling adults: the InCHIANTI Study. AB - AIMS: We examined the relationship between plasma selenium levels at enrollment and all-cause mortality over a 6-year period among participants in the InCHIANTI study. METHODS: 1042 men and women > or =65 years from the InCHIANTI study, a population- based study of older adults living in the Chianti region of Tuscany, a population-based cohort in Tuscany, Italy. Plasma selenium was measured at enrollment (1998-2000), and vital status was ascertained until May 2006. RESULTS: During follow-up, 237 participants (22.7%) died. At enrollment, mean (SD) plasma selenium concentrations among participants who survived or died were 0.96 (0.14) and 0.87 (0.18) micromol/L (p<0.0001), respectively. The proportion of participants who died, from lowest to highest quartile of selenium, was 41.3, 27.0, 18.1 and 13.5% (p<0.0001 by Mantel-Haenszel chi-square). After adjusting for age, sex, education, and chronic diseases, adults in the lowest quartile of plasma selenium at enrollment had higher mortality compared with those in the highest quartile (Hazard Ratio (HR) 1.60, 95% Confidence Interval (CI) 1.04-2.47, p=0.034). CONCLUSION: Low plasma selenium may be an independent predictor of mortality among older adults living in the community. PMID- 18431084 TI - Effects of a physical and nutritional intervention program for frail elderly people over age 75. A randomized controlled pilot treatment trial. AB - BACKGROUND AND AIMS: There are few studies published that combine the interventions of physical training and nutrition. The aim of the present study was to describe the impact of a physical and nutritional intervention program for frail community- dwelling elderly people over the age of 75. METHODS: Ninety-six community-dwelling elderly people (58 women) were randomized to four different groups: i) a physical training program (aerobic, muscle strength, balance), ii) a nutritional intervention program (individually targeted advice and group sessions), iii) a combination of these interventions, and iv) a control group. At baseline subjects were screened for physical performance such as muscle strength, balance, mobility and activities of daily living, as well as nutritional aspects such as energy intake, body weight and fat-free mass. These measurements were repeated immediately after the intervention, which lasted for 12 weeks, and after another 6 months. RESULTS: The intention-to-treat analysis indicated significant improvements in lower- extremity muscle strength in both training groups compared with the nutrition group at 1st follow-up. There were small significant changes for some of the balance measurements in the training group without nutrition treatment. The nutrition intervention did not show any significant results. CONCLUSIONS: This study shows the positive effect on lower-extremity muscle strength directly after the intervention. Balance training most probably needs to be more individualized in order to be effective for frail elderly people. Further studies are needed, with larger sample sizes, to investigate the effects of these types of interventions before any further conclusions can be drawn. PMID- 18431085 TI - Platelet activating factor-acetylhydrolase (PAF-AH) activity and HDL levels, but not PAF-AH gene polymorphisms, are associated with successful aging in Sicilian octogenarians. AB - BACKGROUND AND AIMS: Aging is associated with an increased risk of developing atherosclerosis. Subjects over 80 years of age without cardiovascular disease provide a model to investigate the protective factors increasing their resistance to atherosclerotic disease. Platelet-activating factor acetylhydrolase (PAF-AH) is an enzyme associated with low density lipoprotein (LDL) and high density lipoprotein (HDL) inactivating platelet-activating factor (PAF) and preventing LDL oxidation by hydrolysis of oxidized phospholipids. The aim of the present study was to evaluate the contribution of the PAFAH gene Arg92His, Ile198Thr and Ala379Val polymorphisms to resistance toward developing cardiovascular events in healthy Sicilian octogenarians. METHODS: Distribution of PAF-AH genotypes and activity, and biochemical parameters, were compared between 100 octogenarians and 200 healthy adults. RESULTS: The individuals in the elderly group displayed significantly higher levels of HDL-C (p<0.001) and plasma (p<0.001) and HDL (p<0.001) PAF-AH activity. Analysis of PAFAH genotype distributions showed no significant differences between octogenarians and controls. No differences among PAF-AH genotypes with respect to plasma and HDL PAF-AH activity were found in either group. CONCLUSIONS: Our results provide no evidence of a significant association between the PAF-AH gene Arg92His, Ile198Thr and Ala379Val polymorphisms and successful aging in Sicilians. They also emphasize that, in these subjects, aging is characterized by increased levels of PAF-AH activity and HDL-C. PMID- 18431086 TI - Survivin knockdown combined with apoptin overexpression inhibits cell growth significantly. AB - The initiation and progression of tumor is regulated by multiple genes. Survivin belongs to the inhibitor of apoptosis protein (IAP) family and is overexpressed in most types of human tumors. Apoptin, originally identified from chicken anemia virus (CAV), can specifically induce apoptosis of human tumor cells rather than normal cells. In this study, survivin expression was silenced by microRNA (miRNA) mediated RNA interference (RNAi); meanwhile, the engineered miRNA vector was also designed to express apoptin gene. The apoptosis and cell growth were then examined by flow cytometry and MTT assay. The miRNA-mediated knockdown of survivin in combination with apoptin overexpression significantly induced apoptosis and inhibited cell growth. Importantly, the combined strategy was more effective on inducing apoptosis and inhibiting cell growth than either survivin downregulation or apoptin overexpression alone. Taken together, the combined strategy offers potential advantages in control of tumorigenesis, and thus deserves further research as a preferred approach in cancer gene therapy. PMID- 18431087 TI - Stereomicroscopic fluorescence imaging of head and neck cancer xenografts targeting CD147. AB - PURPOSE: To demonstrate that systemically administered fluorescently labeled anti CD147 antibody can detect head and neck squamous cell carcinoma xenografts in vivo. EXPERIMENTAL DESIGN: In vivo immunodeficient murine model. RESULTS: Peak tumor fluorescence was visualized by near infrared stereomicroscopy in SCC-1 tumors at 24 hours after systemic injection of anti-CD147:Cy5.5 bioconjugate. SCC 1 xenografts demonstrated significantly higher fluorescent intensity after administration of CD147:Cy5.5 (48 au, p < 0.0001) compared to IgG1k:Cy5.5 isotype control antibody (9 au). FaDu tumors overexpressing CD147 (FaDu/E) demonstrated higher fluorescence (53 au) compared to control vector transfected cells (FaDu, 33 au, p < 0.0001) which was higher than CD147 knockdown cells (FaDu/siE, 5 au, p < 0.0001). METHODS: To determine if fluorescently labeled anti-CD147 antibody was specific for tumors in vivo, anti-CD147 and non-specific IgG1k antibody were labeled with a near infrared fluorophore (Cy5.5) and administered systemically to immunodeficient mice bearing SCC-1 xenografts. Imaging was performed over a 72 hour period using brightfield and fluorescent (685-735 nm) stereomicroscopy. To determine if fluorescence varied with receptor expression, SCID mice were xenografted with cell lines expressing variable amounts of CD147: FaDu (control vector transfected), FaDu/siE (siRNA CD147 knockdown) or FaDu/E (CD147 overexpressing) cells. CONCLUSIONS: This data suggests fluorescently labeled anti CD147 may have clinical utility in detection of HNSCC. PMID- 18431088 TI - Dexamethasone reduces tumor recurrence and metastasis after pancreatic tumor resection in SCID mice. AB - BACKGROUND: Glucocorticoids are among the most potent anti-inflammatory agents that act by inhibiting the synthesis of almost all known cytokines and influencing multiple transduction pathways. Inflammation accompanies most solid cancers including pancreatic ductal adenocarcinoma (PDAC), one of the most fatal cancers with surgery being the only potential curative therapeutic. In the present work we investigated the influence of glucocorticoids on PDAC cells in vitro as well as in vivo in a pancreatic carcinoma resection mouse model. METHODS: The influence of dexamethasone (DEX), a synthetic glucocorticoid, on proliferation and IL8 secretion in pancreatic cells (BxPC3, Colo357, PancTuI) was analyzed by cell counting and ELISA. NFkappaB-activity of PancTuI cells treated with DEX was determined by electrophoretic mobility shift assay (EMSA). Furthermore, effects of DEX on the invasiveness were studied by a fibroblast based invasion assay. In the mouse resection model subtotal pancreatectomy was performed after orthotopic inoculation of human PDAC cells. DEX was administered after resection as an adjuvant treatment regime and 4 weeks later, local recurrent tumor sizes as well as number of liver and spleen metastases were analyzed. RESULTS: In vitro, DEX did not have an anti-proliferative effect on PDAC cells, but strongly reduced the invasiveness well as the activation of NFkappaB. The secretion of IL-8 into the supernatant of the tumor cells correlated inversely with the reduced activation of NFkappaB. In vivo, we observed a significant reduction of the local recurrent tumor volume and the number of liver and spleen metastases. CONCLUSIONS: DEX has a profound influence on the malignant phenotype of PDAC tumor cells in vitro in terms of inhibition of invasiveness and pro-inflammatory signaling. This was approved in vivo by reduced metastasizing capacity and reduced size of local tumor recurrence. Therefore, DEX treatment appears to be an interesting therapeutical option in an adjuvant setting after pancreatic cancer resection. PMID- 18431089 TI - Osteoporosis risk factor assessment increases the appropriate use of dual energy X-ray absorptiometry in men and reduces ethnic disparity. AB - BACKGROUND: Male patients are frequently not tested for osteoporosis even in the presence of recognized risk factors for that disease. OBJECTIVES: To evaluate if the assessment of risk factors for osteoporosis increases the utility of dual energy X-ray absorptiometry (DXA) in men over the age of 50 attending a rheumatology clinic. METHODS: Men over 50 attending a rheumatology clinic completed a checklist of 10 risk factors for osteoporosis before seeing the physician. The physician reviewed the checklist and made a management decision. The checklists and medical records were reviewed for medical history and DXA results. Comparisons were made with DXA requests before the use of the checklist. RESULTS: Medical records of 183 men were reviewed, including 111 African Americans (AA) and 67 whites. Twenty-three percent of patients had rheumatoid arthritis (14% of AA, 37% of whites) and 27% of patients were on glucococorticoids. Before the use of the checklist, 14% of men had a DXA (6% of AA and 29% of whites) compared with 29% of patients (21% for AA and 42% for whites) after the checklist was instituted in the clinic. Sixty-three percent of AA with rheumatoid arthritis had DXA compared with 65% of whites. Thirteen patients had osteoporosis whereas 16 had osteopenia. CONCLUSIONS: The use of a check list of risk factors for osteoporosis may increase the appropriate use of DXA in male patients over the age of 50 at risk for osteoporosis. PMID- 18431090 TI - Higher therapeutic plasma oxypurinol concentrations might be required for gouty patients with chronic kidney disease. AB - BACKGROUND: Lower dosages of allopurinol are recommended to avoid toxicity in gout patients with impaired renal function. This often has resulted in inadequate control of hyperuricemia. The optimum therapeutic range of plasma oxypurinol concentrations in gout patients with chronic kidney disease has never been investigated. This study was performed to examine the relationships between plasma oxypurinol concentrations and the changes in serum urate level and renal function after taking a standard dose of allopurinol, 300 mg daily, in gout patients with renal insufficiency. PATIENTS AND METHODS: The study was conducted in 27 gout patients with renal insufficiency in a rheumatology clinic at the Rajvithi Hospital, Bangkok. Both new and current patients, after they discontinued allopurinol completely for 4 weeks, were treated with allopurinol 300 mg daily for 6 weeks. Blood samples were collected immediately before and 5 hours after the studied dose had been taken. Serum urate levels and renal function were recorded before and after the 6 weeks of allopurinol treatments. RESULTS: Most patients receiving allopurinol 300 mg/d had their plasma oxypurinol concentrations higher than the proposed therapeutic range for patients with normal renal function. There were significant relationships between changes in serum urate level with both trough and fifth-hour oxypurinol concentrations (R = 0.42, P = 0.002 and R = 0.27, P = 0.007, respectively). Higher plasma oxypurinol concentrations resulted in a higher percentage of patients who could meet the therapeutic treatment goal. No serious side effect and no significantly change in creatinine clearance were observed indicating that high levels of oxypurinol did not appear to relate to higher prevalence of adverse reaction. CONCLUSIONS: Higher percentages of patients could meet the treatment goal when their plasma oxypurinol concentrations were higher than the proposed therapeutic range for patients with normal renal function. PMID- 18431091 TI - Diffuse musculoskeletal pain is not associated with low vitamin D levels or improved by treatment with vitamin D. AB - BACKGROUND: Uncontrolled studies have suggested that vitamin D insufficiency causes diffuse musculoskeletal pain. OBJECTIVES: Comparison of vitamin D levels in patients with diffuse musculoskeletal pain with controls; evaluation of the effect of treatment with vitamin D on diffuse pain. METHODS: 25-Hydroxyvitamin D levels were measured in patients with diffuse musculoskeletal pain and osteoarthritis (controls) recruited from a community rheumatology practice. The diffuse pain patients with 25-hydroxyvitamin D levels < or = 20 ng/mL were randomized to receive placebo or ergocalciferol 50,000 IU once weekly for 3 months. Outcomes assessed were pain measured by visual analog scale (VAS) and functional pain score (FPS). RESULTS: One hundred eighty-four patients with diffuse pain and 104 with osteoarthritis entered the study. Mean 25 hydroxyvitamin D levels did not differ between the groups (diffuse pain 29.2 ng/mL +/- 13.0, controls 28.8 ng/mL +/- 10.5; P = 0.78); nor did the percent of patients in each group with vitamin D levels < or = 20 ng/mL (diffuse pain 29%, controls 20%; P = 0.09). Fifty patients with diffuse pain who had 25 hydroxyvitamin D levels < or = 20 ng/mL were randomized to receive vitamin D or placebo for 3 months. Vitamin D treatment had no effect on pain in comparison to baseline (VAS P = 0.73; FPS P = 0.18) or at 3 months in comparison to placebo (VAS P = 0.12; FPS P = 0.05, in favor of placebo). CONCLUSIONS: Low vitamin D levels are not associated with diffuse musculoskeletal pain, and treatment with vitamin D does not reduce pain in patients with diffuse pain who have low vitamin D levels. PMID- 18431092 TI - Minocycline-induced hyperpigmentation in rheumatoid arthritis. AB - BACKGROUND: Minocycline is recognized as an effective, well-tolerated therapy in rheumatoid arthritis (RA), although its use has been associated with the development of cutaneous hyperpigmentation. OBJECTIVES: To assess the clinical determinants and frequency of minocycline-induced hyperpigmentation in patients with RA. METHODS: A retrospective medical record review of all patients with RA seen in 2 academic rheumatology practices was performed to identify subjects who had received at least 1 month of continuous minocycline therapy. Patient demographics, disease characteristics, medication use, and medication side effects were abstracted from the medical record. Using Cox proportional hazards regression and restricting the analysis to the initial minocycline course, we examined the association of patient factors and concomitant medications with the development of hyperpigmentation. RESULTS: Of 121 patients with at least 1 minocycline course of 30 days or more, 44 (36%) developed documented hyperpigmentation, including 33 during the initial course over a median duration of 9.1 month (range 2.2-77.8 months). Hyperpigmentation was most commonly seen on the upper and lower extremities and the head/neck region. Minocycline-induced hyperpigmentation led to the discontinuation of treatment in 3 patients, with 12 additional patients receiving a dose reduction. Increasing age was the only clinical determinant significantly associated with hyperpigmentation (HR = 1.04; 95% CI 1.00-1.07, P = 0.04). There were no significant associations of sex, weight, concomitant prednisone, or aspirin use with the development of hyperpigmentation. CONCLUSIONS: Minocycline-induced hyperpigmentation is a common complication seen with minocycline use in the treatment of RA, and seems to increase with age. PMID- 18431093 TI - Atypical arthritis due to combined hereditary hemochromatosis and active hepatitis C. AB - A 51-year-old Caucasian female presented with asymmetric arthritis and a positive rheumatoid factor. She was initially treated for rheumatoid arthritis. However, she had features such as abnormal liver function tests and osteoarthritis in an unusual location, the metacarpophalangeal joint. Further workup revealed that the patient had active hepatitis C and hereditary hemochromatosis. Phlebotomy treatment initiation seemed to be associated with improvement in joint symptoms but, more importantly, may have prevented future risk of cirrhosis and hepatocellular cancer. Treatment for the hepatitis C may also be needed. Clinicians should look for underlying systemic illnesses leading to atypical inflammatory arthritis. PMID- 18431094 TI - Relapsing polychondritis presenting with complete heart block. AB - Relapsing polychondritis (RP) is a rare autoimmune disease characterized by recurrent inflammation of cartilagenous structures, including the ears, nose, trachea, and joints. Cardiovascular involvement is relatively common, but involvement of the conduction system leading to various degrees of heart block is a rare and late manifestation of the disease usually requiring a pacemaker. We report here a patient with RP who presented with complete heart block that resolved promptly after treatment with prednisone. To our knowledge, this is the first reported case of RP with abnormal cardiac conduction on initial presentation and a rapid response to anti-inflammatory treatment. Heart block during the phase of active inflammation may respond to corticosteroid treatment as in our patient without the need for cardiac pacing. PMID- 18431095 TI - Secondary cutaneous nodular AA amyloidosis in a patient with primary Sjogren syndrome and celiac disease. AB - We describe a 62-year-old female with primary Sjogren syndrome and myopathy, severe osteoporosis, and vertebral fractures that were attributed to celiac disease. A year after the diagnosis, she developed a skin nodule on the extensor surface of her right elbow, which was due to an amyloid deposit of AA type. Amyloidosis, although relatively common in some chronic inflammatory diseases, has been uncommon in Sjogren syndrome or celiac disease. Visceral amyloid was not found in this patient. PMID- 18431096 TI - Methylphenidate and dextroamphetamine-induced peripheral vasculopathy. AB - Methylphenidate and dextroamphetamine are central nervous system stimulants used in the treatment of attention deficit hyperactivity disorders in children. These medications have been associated with cerebral arteritis, renal necrotizing vasculitis, and systemic and pulmonary hypertension. We report 4 patients, 2 on methylphenidate and 2 on dextroamphetamine who presented with acral cyanosis, livedo reticularis, or Raynaud phenomenon. Two patients were found to have a positive ANA at low titers, 1 of whom had histopathologic evidence of stratum malgiphian necrosis with perivascular lymphocytic infiltration on skin biopsy. Two had positive antihistone antibodies. One patient improved after withdrawal of dextroamphetamine; others had worsening of their symptoms on higher doses of medications. These cases indicate the potential for development of acral cyanosis, livedo reticularis, or Raynaud symptoms with these medications and their potential contribution to a vasculopathy. PMID- 18431097 TI - An unusual multiplex systemic lupus erythematosus family with high prevalence of nephropathy, late-onset disease, and one member with disease-onset post-HIV therapy. AB - In the present study, we report the clinical characteristics of a unique systemic lupus erythematosus (SLE) multiplex family with 6 of its members affected by the disease, 1 of them being male. Four patients showed nephropathy, 2 of them with late-onset SLE (52 and 55-year-old), one with cutaneous and articular involvement, and another one developing lupus after 5 years undergoing highly active antiretroviral therapy (HAART) due to acquired immunodeficiency syndrome. Notwithstanding the genetic load, the fact that 2 patients showed late-onset disease, and the extreme delay of the appearance of SLE after HAART in the proband suggest that not only genetic, but other--mainly environmental--factors are necessarily required for the development of SLE. PMID- 18431098 TI - Dual ANCA positivity in subacute bacterial endocarditis. AB - Nine cases of subacute bacterial endocarditis (SBE) associated with anti-PR3 ANCA have been described in the literature to date. We describe 2 cases of SBE associated with dual ANCA positivity (anti-PR3 and anti-MPO ANCA.) To our knowledge, these are the first such reported cases. One case was associated with cutaneous vasculitis, and the second with predisposing factors for SBE. ANCA titers resolved or decreased in both during initial corticosteroid therapy for suspected vasculitis. Follow-up of these patients revealed no evidence of the common ANCA associations such as Wegener granulomatosis. Rheumatologists, if aware of ANCA association with SBE, can avoid inappropriate immunosuppressive treatments. PMID- 18431099 TI - Theodore Roosevelt's inflammatory rheumatism. AB - Theodore Roosevelt's death at age 60 was probably caused by a pulmonary embolus, but it was preceded by a 2 1/2-month illness described as inflammatory rheumatism. He had intermittent fever and acute arthritis in several joints leading to hospitalization and enforced bed rest for 6 weeks. The details of his illness and its possible etiology are reviewed. Inflammatory rheumatism was a descriptive term within which several modern diagnoses might be included. Although it is not possible to identify Roosevelt's illness with any certainty, it was most compatible with polyarticular gout, although reactive arthritis, rheumatic fever, and several other diagnoses cannot be ruled out. PMID- 18431100 TI - Basic science for the clinician 48: tyrosine kinases in disease: the potential for inhibitors in the treatment of immunologic diseases. AB - The magic bullet--a compound that will stop a disease dead in its tracks by specifically targeting the underlying pathogenic principle of that disease--is what every designer/developer of drugs wants. As cellular and molecular biology research delves deeper into how cells are activated by their ligands, the intracellular pathways of activation of individual cellular processes become better known and more attractive therapeutic targets. The receptors for transforming growth factor beta (TGFbeta) and platelet-derived growth factor (PDGF) activate a variety of cells via a series of tyrosine kinases; inhibition of specific tyrosine kinases has until recently been within the domain of oncologists, treating leukemia, and certain gastrointestinal tumors, but now there is mounting evidence that these agents might be of value in rheumatologic and autoimmune diseases. This is another example of "Better living (and curing!) through chemistry" that we as clinicians need to master to render optimal care. PMID- 18431102 TI - HLA-B27 and its pathogenic role. PMID- 18431103 TI - Vasculopathy associated with gout. PMID- 18431104 TI - Primary central nervous system lymphoma in rheumatoid arthritis. PMID- 18431105 TI - Candida parapsilosis septic arthritis in a renal transplant patient. PMID- 18431106 TI - Necrotizing cutaneous vasculitis in multibacillary leprosy disease (Lucio's phenomenon). PMID- 18431107 TI - Calcific tendonitis of the rectus femoris. PMID- 18431108 TI - Polymyalgia rheumatica and elderly onset rheumatoid arthritis. PMID- 18431110 TI - Is it more informative to focus on cardiac output than blood pressure during spinal anesthesia for cesarean delivery in women with severe preeclampsia? PMID- 18431111 TI - Pressure wave analysis is useful to understand the pathophysiology of preeclampsia, but perhaps not the rapid changes during cesarean delivery. PMID- 18431112 TI - An optic nerve at risk and a prolonged surgery in the prone position: time to reconsider? PMID- 18431113 TI - Gerard W. Ostheimer "What's new in obstetric anesthesia" lecture. PMID- 18431114 TI - Practice advisory for the prevention and management of operating room fires. PMID- 18431115 TI - Hemodynamic changes associated with spinal anesthesia for cesarean delivery in severe preeclampsia. AB - BACKGROUND: Hemodynamic responses to spinal anesthesia (SA) for cesarean delivery in patients with severe preeclampsia are poorly understood. This study used a beat-by-beat monitor of cardiac output (CO) to characterize the response to SA. The hypothesis was that CO would decrease from baseline values by less than 20%. METHODS: Fifteen patients with severe preeclampsia consented to an observational study. The monitor employed used pulse wave form analysis to estimate nominal stroke volume. Calibration was by lithium dilution. CO and systemic vascular resistance were derived from the measured stroke volume, heart rate, and mean arterial pressure. In addition, the hemodynamic effects of phenylephrine, the response to delivery and oxytocin, and hemodynamics during recovery from SA were recorded. Hemodynamic values were averaged for defined time intervals before, during, and after SA. RESULTS: Cardiac output remained stable from induction of SA until the time of request for analgesia. Mean arterial pressure and systemic vascular resistance decreased significantly from the time of adoption of the supine position until the end of surgery. After oxytocin administration, systemic vascular resistance decreased and heart rate and CO increased. Phenylephrine, 50 mug, increased mean arterial pressure to above target values and did not significantly change CO. At the time of recovery from SA, there were no clinically relevant changes from baseline hemodynamic values. CONCLUSIONS: Spinal anesthesia in severe preeclampsia was associated with clinically insignificant changes in CO. Phenylephrine restored mean arterial pressure but did not increase maternal CO. Oxytocin caused transient marked hypotension, tachycardia, and increases in CO. PMID- 18431116 TI - STOP questionnaire: a tool to screen patients for obstructive sleep apnea. AB - BACKGROUND: Obstructive sleep apnea (OSA) is a major risk factor for perioperative adverse events. However, no screening tool for OSA has been validated in surgical patients. This study was conducted to develop and validate a concise and easy-to-use questionnaire for OSA screening in surgical patients. METHODS: After hospital ethics approval, preoperative patients aged 18 yr or older and without previously diagnosed OSA were recruited. After a factor analysis, reliability check, and pilot study; four yes/no questions were used to develop this screening tool. The four questions were respectively related to snoring, tiredness during daytime, observed apnea, and high blood pressure (STOP). For validation, the score from the STOP questionnaire was evaluated versus the apnea-hypopnea index from monitored polysomnography. RESULTS: The STOP questionnaire was given to 2,467 patients, 27.5% classified as being at high risk of OSA. Two hundred eleven patients underwent polysomnography, 34 for the pilot test and 177 for validation. In the validation group, the apnea-hypopnea index was 20 +/- 6. The sensitivities of the STOP questionnaire with apnea-hypopnea index greater than 5, greater than 15, and greater than 30 as cutoffs were 65.6, 74.3, and 79.5%, respectively. When incorporating body mass index, age, neck circumference, and gender into the STOP questionnaire, sensitivities were increased to 83.6, 92.9, and 100% with the same apnea-hypopnea index cutoffs. CONCLUSIONS: The STOP questionnaire is a concise and easy-to-use screening tool for OSA. It has been developed and validated in surgical patients at preoperative clinics. Combined with body mass index, age, neck size, and gender, it had a high sensitivity, especially for patients with moderate to severe OSA. PMID- 18431117 TI - Validation of the Berlin questionnaire and American Society of Anesthesiologists checklist as screening tools for obstructive sleep apnea in surgical patients. AB - BACKGROUND: Because of the high prevalence of obstructive sleep apnea (OSA) and its adverse impact on perioperative outcome, a practical screening tool for surgical patients is required. This study was conducted to validate the Berlin questionnaire and the American Society of Anesthesiologists (ASA) checklist in surgical patients and to compare them with the STOP questionnaire. METHODS: After hospital ethics approval, preoperative patients aged 18 yr or older and without previously diagnosed OSA were recruited. The scores from the Berlin questionnaire, ASA checklist, and STOP questionnaire were evaluated versus the apnea-hypopnea index from in-laboratory polysomnography. The perioperative data were collected through chart review. RESULTS: Of 2,467 screened patients, 33, 27, and 28% were respectively classified as being at high risk of OSA by the Berlin questionnaire, ASA checklist, and STOP questionnaire. The performance of the screening tools was evaluated in 177 patients who underwent polysomnography. The sensitivities of the Berlin questionnaire, ASA checklist, and STOP questionnaire were 68.9-87.2, 72.1-87.2, and 65.6-79.5% at different apnea-hypopnea index cutoffs. There was no significant difference between the three screening tools in the predictive parameters. The patients with an apnea-hypopnea index greater than 5 and the patients identified as being at high risk of OSA by the STOP questionnaire or ASA checklist had a significantly increased incidence of postoperative complications. CONCLUSIONS: Similar to the STOP questionnaire, the Berlin questionnaire and ASA checklist demonstrated a moderately high level of sensitivity for OSA screening. The STOP questionnaire and the ASA checklist were able to identify the patients who were likely to develop postoperative complications. PMID- 18431118 TI - Part Task and variable priority training in first-year anesthesia resident education: a combined didactic and simulation-based approach to improve management of adverse airway and respiratory events. AB - BACKGROUND: Part task training (PTT) focuses on dividing complex tasks into components followed by intensive concentrated training on individual components. Variable priority training (VPT) focuses on optimal distribution of attention when performing multiple tasks simultaneously with the goal of flexible allocation of attention. This study explored how principles of PTT and VPT adapted to anesthesia training would improve first-year anesthesiology residents' management of simulated adverse airway and respiratory events. The authors hypothesized that participants with PTT and VPT would perform better than those with standard training. METHODS: Twenty-two first-year anesthesia residents were randomly divided into two groups and trained over 12 months. The control group received standard didactic and simulation-based training. The experimental group received similar training but with emphasis on PTT and VPT techniques. Participant ability to manage seven adverse airway and respiratory events were assessed before and after the training period. Performance was measured by the number of correct tasks, making a correct diagnosis, assessment of perceived workload, and an assessment of scenario comprehension. RESULTS: Participants in both groups exhibited significant improvement in all metrics after a year of training. Participants in the experimental group were able to complete more tasks and answered more comprehension questions correctly. There was no difference in perceived workload or the number of correct diagnoses between groups. CONCLUSION: This study in part confirmed the study hypotheses. The results suggest that VPT and PTT are promising adjuncts to didactic and simulation-based training for management of adverse airway and respiratory events. PMID- 18431119 TI - Anesthesia depth-dependent features of electroencephalographic bicoherence spectrum during sevoflurane anesthesia. AB - BACKGROUND: Growth pattern in the electroencephalographic bicoherence spectrum has recently been found to relate to anesthetic depth, and bicoherence analysis can reflect behavior of the thalamocortical reverberating network. Because the thalamocortical network is known to represent a key factor in sleep by anesthesia, systematic and qualitative bicoherence studies of different anesthetic depths is necessary throughout all pairs of frequencies. METHODS: Sixteen patients were anesthetized using sevoflurane (1, 2, or 3%) combined with remifentanil (0.4 microg x kg x min). Raw electroencephalographic signals were collected, and bicoherence was estimated in all pairs of frequencies, between 0.5 and 40 Hz at 0.5-Hz intervals. RESULTS: Sevoflurane (1%) caused two main peaks, spindle frequencies (11.0 +/- 1.2 Hz, 44.7 +/- 12.3% [bicoherence growth]) and delta-theta frequencies (5.4 +/- 0.5 Hz, 33.0 +/- 8.4%), in the diagonal line of biphasic bicoherence plots. High concentrations of sevoflurane (2% and 3%) shifted these peaks to 9.8 +/- 1.1 Hz, 46.2 +/- 12.7%; 8.7 +/- 1.3 Hz, 37.2 +/- 13.7% and 4.9 +/- 0.5 Hz, 44.6 +/- 7.0%; 4.3 +/- 0.8 Hz, 45.2 +/- 10.6%, respectively. Sevoflurane caused a third bicoherence peak to appear in another heterogeneous pair frequency (pair of alpha basal frequency and its double frequency), outside the diagonal line, which also inherited the behavior of alpha bicoherence peaks at different anesthetic depths. CONCLUSIONS: Sevoflurane anesthesia caused bicoherence peaks in alpha and delta-theta areas and also formed secondary third peaks. Deeper sevoflurane anesthesia shifted all bicoherence peaks to lower frequencies and caused increased bicoherence growth in the delta-theta area. The obtained features are consistent with characteristics of the thalamocortical reverberating network and suggest the importance of bicoherence analysis for the thalamic system. PMID- 18431120 TI - Comparison of bispectral index and composite auditory evoked potential index for monitoring depth of hypnosis in children. AB - BACKGROUND: In pediatric patients, the Bispectral Index (BIS), derived from the electroencephalogram, and the composite A-Line autoregressive index (cAAI), derived from auditory evoked potentials and the electroencephalogram, have been used as measurements of depth of hypnosis during anesthesia. The performance and reliability of BIS and cAAI in distinguishing different hypnotic states in children, as evaluated with the University of Michigan Sedation Scale, were compared. METHODS: Thirty-nine children (aged 2-16 yr) scheduled to undergo elective inguinal hernia surgery were studied. For all patients, standardized anesthesia was used. Prediction probabilities of BIS and cAAI versus the University of Michigan Sedation Scale and sensitivity/specificity were calculated. RESULTS: Prediction probabilities for BIS and cAAI during induction were 0.84 for both and during emergence were 0.75 and 0.74, respectively. At loss of consciousness, the median BIS remained unaltered (94 to 90; not significant), whereas cAAI values decreased (60 to 43; P < 0.001). During emergence, median BIS and cAAI increased from 51 to 74 (P < 0.003) and from 46 to 58 (P < 0.001), respectively. With respect to indicate consciousness or unconsciousness, 100% sensitivity was reached at cutoff values of 17 for BIS and 12 for cAAI. One hundred percent specificity was associated with a BIS of 71 and a cAAI of 60. To ascertain consciousness, BIS values greater than 78 and cAAI values above 52 were required. CONCLUSIONS: BIS and cAAI were comparable indicators of depth of hypnosis in children. Both indices, however, showed considerable overlap for different clinical conditions. PMID- 18431121 TI - Sevoflurane concentrations required to block autonomic hyperreflexia during transurethral litholapaxy in patients with complete spinal cord injury. AB - BACKGROUND: Autonomic hyperreflexia (AHR) is a potentially life-threatening hypertensive condition that occurs in patients with high spinal cord injury (SCI). The current study was aimed to determine sevoflurane concentrations that block AHR in SCI patients. METHODS: The study involved 28 patients with chronic, complete SCI scheduled to undergo transurethral litholapaxy during general anesthesia. Nine patients without SCI served as controls post hoc. Anesthesia was induced with thiopental, and sevoflurane concentrations in 50% nitrous oxide were adjusted to maintain a Bispectral Index of 40-50. When a patient developed AHR during bladder distension, the target sevoflurane concentration was maintained for at least 10 min, and then the procedure was resumed. Systolic blood pressure, heart rate, and Bispectral Index as well as plasma concentrations of catecholamines and arginine vasopressin were measured before and during the bladder distension. Each target concentration was determined by the up-and-down method based on changes (15% increase or more) of systolic blood pressure in response to bladder distension. RESULTS: In SCI, systolic pressure increased by 67 +/- 33 mmHg, whereas heart rate decreased by 13 +/- 8 beats/min during the first trial (P < 0.01). The hypertensive event was associated with increases of norepinephrine concentrations, but not of epinephrine or vasopressin concentrations. Systolic pressure, heart rate, and norepinephrine concentrations did not change significantly in the control patients. The end-tidal concentrations of sevoflurane to prevent AHR were EC50 of 3.12% and EC95 of 3.83%. CONCLUSION: The EC95 for sevoflurane in 50% nitrous oxide to block AHR during transurethral litholapaxy in patients with SCI was 3.83%. PMID- 18431122 TI - Effects of anemia and hypotension on porcine optic nerve blood flow and oxygen delivery. AB - BACKGROUND: Perioperative ischemic optic neuropathy occurs after major surgical procedures, which are often associated with hypotension, anemia, or venous congestion. However, the effects of these conditions on optic nerve (ON) blood flow are unknown and cannot be studied adequately in humans. METHODS: Farm-raised pigs were anesthetized with isoflurane, kept normocapnic and normothermic, and subjected to conditions of euvolemic or hypovolemic hypotension (mean arterial pressure 50-55 mm Hg), anemia (hematocrit 17%), venous congestion, and combinations thereof. Control animals were kept euvolemic and normotensive for the entire experiment. Fluorescent microspheres were used to measure cerebral blood flow (CBF) and ON blood flow at baseline and after experimental conditions, and to calculate oxygen delivery (DO2). RESULTS: No significant changes in CBF or ON blood flow or DO2 occurred with euvolemic hypotension (n = 5), compared with controls (n = 12). Hypovolemic hypotension (n = 4) resulted in stable CBF and cerebral DO2, but significant reductions in ON DO2 (P = 0.032). The significant increase in CBF associated with anemia (n = 6) resulted in stable cerebral DO2. In contrast, ON blood flow did not significantly change with anemia, with (n = 5) or without (n = 6) euvolemic hypotension, resulting in significant reductions in ON DO2 (P < 0.01). CONCLUSION: Compensatory mechanisms for porcine CBF maintain stable DO2 under specified conditions of hypotension or anemia, whereas ON compensatory mechanisms were unable to maintain blood flow and to preserve DO2. The authors conclude that the porcine ON is more susceptible to physiologic perturbations than the brain. PMID- 18431123 TI - Hypertonic resuscitation improves neuronal and behavioral outcomes after traumatic brain injury plus hemorrhage. AB - BACKGROUND: : Resuscitation with hypertonic saline or hypertonic saline plus l arginine acutely improves cerebral blood flow after traumatic brain injury (TBI) followed by hemorrhagic hypotension. The authors investigated whether hypertonic saline or hypertonic l-arginine would improve long-term neuronal survival and behavioral outcomes 15 days after TBI and hemorrhagic hypotension. METHODS: : Mean arterial pressure, arterial blood gases, pH, plasma glucose, hematocrit, and hemoglobin were measured in male Sprague-Dawley rats before and after moderate (2.0 atm) fluid percussion TBI. Rats were assigned to one of six groups: (1) sham TBI, (2) hemorrhage only, (3) TBI only, (4) TBI plus hemorrhage and resuscitation with 0.9% saline, (5) TBI plus hemorrhage and resuscitation with hypertonic saline (7.5%), or (6) TBI plus hemorrhage and resuscitation with l-arginine (100 mg/kg) in hypertonic saline. On postinjury days 1-5, vestibulomotor function was assessed using beam balance and beam walking tasks. On postinjury days 11-15, spatial memory function was assessed using the Morris water maze. After behavioral testing, neuronal counting was performed bilaterally on specific hippocampal regions. RESULTS: : Groups receiving hypertonic saline (P < 0.05, day 15 vs. day 11) or hypertonic l-arginine (P < 0.05, days 13-15 vs. day 11) showed improved performance over time on the Morris water maze, as well as significantly improved neuronal survival in the contralateral hippocampus (P < 0.05, hypertonic saline or hypertonic l-arginine vs. normal saline) compared with untreated TBI or normal saline-treated TBI plus hemorrhage groups. CONCLUSIONS: : Hypertonic saline and hypertonic l-arginine were both effective at promoting long-term neuronal survival and behavioral recovery. The slightly earlier improvement in Morris water maze performance in the hypertonic l-arginine group warrants further studies to determine whether higher doses of l-arginine provide additional improvement. This study supports the therapeutic benefits of hypertonic resuscitation after TBI plus hemorrhagic hypotension. PMID- 18431124 TI - Effect of disrupting N-methyl-d-aspartate receptor-postsynaptic density protein 95 interactions on the threshold for halothane anesthesia in mice. AB - BACKGROUND: The authors' previous studies have shown that clinically relevant concentrations of inhalational anesthetics dose-dependently and specifically inhibit the PSD-95, Dlg, and ZO-1 (PDZ) domain-mediated protein interactions between postsynaptic density protein 95 (PSD-95) and N-methyl-d-aspartate receptors, and that the knockdown of spinal PSD-95 by intrathecal injection of PSD-95 antisense oligodeoxynucleotide significantly reduces the minimum alveolar anesthetic concentration for isoflurane in rats. METHODS: The authors constructed a fusion peptide, Tat-PSD-95 PDZ2, comprising the second PDZ domain of PSD-95, which can specifically disrupt PSD-95 PDZ2-mediated protein interactions by binding to its interaction partner. By intraperitoneal injection of this fusion peptide into mice, the authors investigated the effect of disrupting the PSD-95 PDZ2-mediated protein interactions on the threshold for halothane anesthesia. RESULTS: Systemically injected fusion peptide Tat-PSD-95 PDZ2 was delivered into the central nervous system, disrupted the protein-protein interactions between N methyl-d-aspartate receptor NR2 subunits and PSD-95, and significantly reduced the minimum alveolar anesthetic concentration and righting reflex EC50 for halothane. CONCLUSIONS: By disrupting PSD-95 PDZ2 domain-mediated protein interactions, intraperitoneal injection of cell-permeant fusion peptide Tat-PSD 95 PDZ2 dose-dependently reduces the threshold for halothane anesthesia. These results suggest that PDZ domain-mediated protein interactions at synapses in the central nervous system might play an important role in the molecular mechanisms of halothane anesthesia. PMID- 18431125 TI - Involvement of the basal cholinergic forebrain in the mediation of general (propofol) anesthesia. AB - BACKGROUND: Recent studies have pointed out the involvement of the basal forebrain gamma-aminobutyric acid-mediated system in mediating the effects of general anesthesia. In this study, the authors asked whether the basal forebrain cholinergic system is also involved in mediating the effects of general anesthetics such as propofol. METHODS: Cholinergic lesions were produced by administration of the selective immunotoxin 192 immunoglobulin G-saporin into the lateral ventricles, the medial septum, or the nucleus basalis magnocellularis. The anesthetic potency of propofol was determined using an anesthetic score with a crossover counterbalanced design. Animals were given intraperitoneal propofol (25 or 50 mg/kg) repeatedly every 15 min to set up a subanesthetic (low-dose) or anesthetic (high-dose) state. The anesthetic score was assessed for each cumulative dose. Control of the cholinergic depletion was performed using histochemical acetylcholinesterase staining on brain slices. RESULTS: A shift from a subanesthetic state to an anesthetic state was observed mainly in the rats with the immunotoxin injected into the lateral ventricles or the medial septum and vertical diagonal band of Broca, compared with controls. In those rats, the density of acetylcholinesterase reaction products was normal in the striatum and the thalamus, but reduced in the cortex and the hippocampus. CONCLUSION: The anesthetic potency of propofol was increased in all rats with hippocampal lesions, whatever the injection sites, compared with controls. These results demonstrate that a cholinergic dysfunction in the basal forebrain potentiates the anesthetic effects of propofol. PMID- 18431128 TI - Experience is not enough: repeated breaches in epidural anesthesia aseptic technique by novice operators despite improved skill. AB - BACKGROUND: Invasive procedures such as epidural anesthesia carry risks for complications such as erroneous placement arising from inadequate manual skills and infection secondary to breaches in aseptic technique. Although it is assumed that improvement in aseptic technique parallels improved dexterity, this assertion remains unproven. The aim of this study was to determine whether increased proficiency in the manual skills for epidural anesthesia is associated with improved aseptic technique. METHODS: Second-year anesthesia residents were repeatedly videotaped performing epidural anesthesia over 6-month periods. Three independent examiners blinded to the level of training of the residents evaluated the procedures for manual skills and aseptic technique. Each procedure was graded using a manual skills checklist, a global rating scale, and an aseptic technique checklist. The main outcome measures were the scores for these three tools. RESULTS: Thirty-five sessions were videotaped over 1 yr. Interrater reliability was nearly perfect. A strong positive association was found between increased experience and manual skills, as reflected by the scores achieved on both the manual skills checklist and the global rating scale. In contrast, a nonsignificant or very weak correlation was found between the aseptic technique checklist total scores and the number of epidurals performed. CONCLUSION: Manual skills for invasive procedures improved with increasing experience, but aseptic technique did not, despite formal teaching. These findings reflect major gaps in the understanding and teaching of the principles of aseptic technique, most likely due to lack of structured training. Educational initiatives are needed to correct these teaching gaps. PMID- 18431127 TI - Resuscitation with lipid versus epinephrine in a rat model of bupivacaine overdose. AB - BACKGROUND: Lipid emulsion infusion reverses cardiovascular compromise due to local anesthetic overdose in laboratory and clinical settings. The authors compared resuscitation with lipid, epinephrine, and saline control in a rat model of bupivacaine-induced cardiac toxicity to determine whether lipid provides a benefit over epinephrine. METHODS: Bupivacaine, 20 mg/kg, was infused in rats anesthetized with isoflurane, producing asystole in all subjects. Ventilation with 100% oxygen and chest compressions were begun immediately, along with intravenous treatment with 30% lipid emulsion or saline (5-ml/kg bolus plus continuous infusion at 0.5 ml . kg . min) or epinephrine (30 microg/kg). Chest compressions were continued and boluses were repeated at 2.5 and 5 min until the native rate-pressure product was greater than 20% baseline. Electrocardiogram and arterial pressure were monitored continuously and at 10 min, arterial blood gas, central venous oxygen saturation, and blood lactate were measured. Effect size (Cohen d) was determined for comparisons at 10 min. RESULTS: Lipid infusion resulted in higher rate-pressure product (P < 0.001, d = 3.84), pH (P < 0.01, d = 3.78), arterial oxygen tension (P < 0.05, d = 2.8), and central venous oxygen saturation (P < 0.001, d = 4.9) at 10 min than did epinephrine. Epinephrine treatment caused higher lactate (P < 0.01, d = 1.48), persistent ventricular ectopy in all subjects, pulmonary edema in four of five rats, hypoxemia, and a mixed metabolic and respiratory acidosis by 10 min. CONCLUSIONS: Hemodynamic and metabolic metrics during resuscitation with lipid surpassed those with epinephrine, which were no better than those seen in the saline control group. Further studies are required to optimize the clinical management of systemic local anesthetic toxicity. PMID- 18431126 TI - Differential effects of isoflurane and propofol on upper airway dilator muscle activity and breathing. AB - BACKGROUND: Anesthesia impairs upper airway integrity, but recent data suggest that low doses of some anesthetics increase upper airway dilator muscle activity, an apparent paradox. The authors sought to understand which anesthetics increase or decrease upper airway dilator muscle activity and to study the mechanisms mediating the effect. METHODS: The authors recorded genioglossus electromyogram, breathing, arterial blood pressure, and expiratory carbon dioxide in 58 spontaneously breathing rats at an estimated ED50 (median effective dose) of isoflurane or propofol. The authors further evaluated the dose-response relations of isoflurane under different study conditions: (1) normalization of mean arterial pressure, or end-expiratory carbon dioxide; (2) bilateral lesion of the Kolliker-Fuse nucleus; and (3) vagotomy. To evaluate whether the markedly lower inspiratory genioglossus activity during propofol could be recovered by increasing flow rate, a measure of respiratory drive, the authors performed an additional set of experiments during hypoxia or hypercapnia. RESULTS: In vagally intact rats, tonic and phasic genioglossus activity were markedly higher with isoflurane compared with propofol. Both anesthetics abolished the genioglossus negative pressure reflex. Inspiratory flow rate and anesthetic agent predicted independently phasic genioglossus activity. Isoflurane dose-dependently decreased tonic and increased phasic genioglossus activity, and increased flow rate, and its increasing effects were abolished after vagotomy. Impairment of phasic genioglossus activity during propofol anesthesia was reversed during evoked increase in respiratory drive. CONCLUSION: Isoflurane compared with propofol anesthesia yields higher tonic and phasic genioglossus muscle activity. The level of respiratory depression rather than the level of effective anesthesia correlates closely with the airway dilator muscle function during anesthesia. PMID- 18431129 TI - Local myotoxicity from sustained release of bupivacaine from microparticles. AB - BACKGROUND: Sustained release of local anesthetics is frequently associated with myotoxicity. The authors investigated the role of particulate delivery systems and of the pattern of drug release in causing myotoxicity. METHODS: Rats were given sciatic nerve blocks with bupivacaine solutions, two types of bupivacaine containing microparticles (polymeric microspheres and lipid-protein-sugar particles), or blank particles with or without bupivacaine in the carrier fluid. Myotoxicity was scored in histologic sections of the injection sites. Bupivacaine release kinetics from the particles were measured. Myotoxicity of a range of bupivacaine concentrations from exposures up to 3 weeks was assessed in C2C12 myotubes, with or without microparticles. RESULTS: Both types of bupivacaine loaded microparticles, but not blank particles, were associated with myotoxicity. Whereas 0.5% bupivacaine solution caused little myotoxicity, a concentration of bupivacaine that mimicked the amount of bupivacaine released initially from particles caused myotoxicity. Local anesthetics showed both concentration and time-dependent myotoxicity in C2C12s. Importantly, even very low concentrations that were nontoxic over brief exposures became highly toxic after days or weeks of exposure. The presence of particles did not increase bupivacaine myotoxicity in vitro but did in vivo. Findings applied to both particle types. CONCLUSIONS: Whereas the release vehicles themselves were not myotoxic, both burst and extended release of bupivacaine were. A possible implication of the latter finding is that myotoxicity is an inevitable concomitant of sustained release of local anesthetics. Particles, and perhaps other vehicles, may enhance local toxicity through indirect mechanisms. PMID- 18431130 TI - Glycine transporter inhibitors as a potential therapeutic strategy for chronic pain with memory impairment. AB - BACKGROUND: Impaired excitatory and inhibitory balance in the spinal dorsal horn has a crucial role in the pathophysiology of chronic pain. The authors addressed the therapeutic impact of increasing spinal glycine applied exogenously or via blockade of glycine transporter 1 using its selective inhibitors sarcosine and N [3-(4'-fluorophenyl)-3-(4'-phenylphenoxy)propyl]sarcosine on neuropathic and inflammatory pain in mice. METHODS: Mice with thermal and mechanical hypersensitivity after partial ligation of the sciatic nerve (Seltzer model) or mice with mechanical hypersensitivity after streptozotocin injection received intrathecal injection of glycine, sarcosine, and N-[3-(4'-fluorophenyl)-3-(4' phenylphenoxy)propyl]sarcosine. These drugs were also intrathecally injected in mice to assess their effects on formalin-evoked nociceptive behaviors. The supraspinal effect of blockade of glycine transporter 1 was studied on tetanus induced long-term potentiation of the Schaffer-collateral synapses in hippocampal slices prepared from Seltzer model mice. RESULTS: Glycine, sarcosine, and N-[3 (4'-fluorophenyl)-3-(4'-phenylphenoxy)propyl]sarcosine ameliorated thermal and mechanical hypersensitivity in Seltzer model mice, and reduced mechanical hypersensitivity in streptozotocin-injected diabetic mice. Moreover, they selectively inhibited the second phase of formalin-evoked licking/biting behavior. In hippocampal slices prepared from Seltzer model mice, long-term potentiation was maintained at a significantly lower level than that in sham treated mice. Such impairment of long-term potentiation was never observed when it was induced in the presence of N-[3-(4'-fluorophenyl)-3-(4' phenylphenoxy)propyl]sarcosine. CONCLUSIONS: An increase in endogenous glycine via glycine transporter 1 blockade not only results in a net inhibitory influence on pain transmission at the spinal level but also supraspinally relieves decreased synaptic efficacy presumably related to cognitive disturbance often described in patients with chronic pain. PMID- 18431131 TI - Toxicology profile of N-methyl-D-aspartate antagonists delivered by intrathecal infusion in the canine model. AB - BACKGROUND: Intrathecal N-methyl-d-aspartate antagonists have antihyperalgesic efficacy. The authors examined toxicity in a canine model of chronic lumbar intrathecal infusion. METHODS: Dogs (10-16 kg) were prepared with lumbar intrathecal catheters connected to vest-mounted pumps (100 microl/h). In phase 1, stepwise incrementations in infusion concentration were performed at 48- to 72-h intervals to determine an infusion dose with minimal but detectable behavioral effects. In phase 2, the dose/concentration defined in phase 1 was infused for 28 days. Behavioral function during infusion and histopathology at sacrifice was assessed. Drugs examined were 2-amino-5-phosphono-valorate (AP5), MK801, memantine, amitriptyline, S-methadone, and saline. RESULTS: In the phase 1 dose ranging, the minimum effect doses for the several agents were as follows: AP5, 1 mg/day; amitriptyline, 1 mg/day; ketamine, 10 mg/day; MK801, 1 mg/day; and memantine, 4 mg/day. In phase 2, infusion of these doses typically resulted in mild hind limb weakness by 3-5 days after initiation of infusion, which progressed over the 28-day infusion interval. In a limited number of animals, a similar effect was observed with S-methadone. Histopathologically, vehicle infused animals displayed a minor local catheter reaction. With the drug treatments, a gradient of increasing pathology from cervical to lumbar segments was noted. Pathology ranged from local demyelination to necrotizing lesions of spinal parenchyma near the catheter tip. All drugs given at their respective doses produced pathology scores significantly worse than saline controls. CONCLUSIONS: These drugs given for 28 days at acutely tolerable doses lead to spinal pathology. These data suggest a reevaluation of the use of these agents in chronic spinal delivery. PMID- 18431132 TI - Utility of the photoplethysmogram in circulatory monitoring. AB - The photoplethysmogram is a noninvasive circulatory signal related to the pulsatile volume of blood in tissue and is displayed by many pulse oximeters and bedside monitors, along with the computed arterial oxygen saturation. The photoplethysmogram is similar in appearance to an arterial blood pressure waveform. Because the former is noninvasive and nearly ubiquitous in hospitals whereas the latter requires invasive measurement, the extraction of circulatory information from the photoplethysmogram has been a popular subject of contemporary research. The photoplethysmogram is a function of the underlying circulation, but the relation is complicated by optical, biomechanical, and physiologic covariates that affect the appearance of the photoplethysmogram. Overall, the photoplethysmogram provides a wealth of circulatory information, but its complex etiology may be a limitation in some novel applications. PMID- 18431133 TI - Extracorporeal cardiopulmonary support for cardiogenic shock caused by pheochromocytoma: a case report and literature review. PMID- 18431134 TI - Cortical electroencephalogram from subcortical electrodes rather than electrosubcorticogram. PMID- 18431137 TI - Anticholinesterase drugs and the transplanted heart. PMID- 18431139 TI - The black box warning for droperidol. PMID- 18431140 TI - Is there a surgical role for topical aprotinin? PMID- 18431142 TI - Differentiated thyroid carcinoma in a juvenile patient. AB - Juvenile differentiated carcinoma thyroid is a rare entity. It differs from adult differentiated thyroid carcinoma in a variety of ways, including large tumor volume at presentation with early involvement of the capsule, more frequent nodal and distant metastases, greater expression of sodium-iodide symporter and early recurrence. The overall survival seems to be better than for adult patients; however, due to high and early recurrence rates, prompt and adequate treatment is advocated. The mainstay of treatment includes total thyroidectomy, central lymphadenectomy with modified radical lateral lymphadenectomy, followed by ablation with radioactive iodine (RAI). Both modalities improve the final outcome, but RAI ablation decreases cause-specific death risk independent of the extent of surgery. We present the case of a 5-year-old girl, the youngest ever treated in our country with surgery and RAI therapy successfully after being diagnosed as papillary carcinoma of the thyroid, follicular variant. PMID- 18431143 TI - F-18 FDG PET-CT findings of intraperitoneal carbon particles-induced granulomas mimicking peritoneal carcinomatosis. AB - F-18 FDG PET-CT is a useful modality for monitoring residual or recurrent tumors after surgical resection. We report on 3 patients with intraperitoneal charcoal induced granulomas mimicking peritoneal carcinomatosis on PET-CT images. Two of them underwent a radical gastrectomy because of advanced gastric cancer, and the other underwent a hemicolectomy because of sigmoid colon cancer. All 3 patients had a history of intraperitoneal chemotherapy using mitomycin C bound to activated carbon particles during surgery. Follow-up PET-CT studies demonstrated increased FDG uptake mimicking peritoneal carcinomatosis on PET images alone. However, the accompanying noncontrast CT showed variously shaped hyperdense nodules in the dependent positions of the peritoneal cavity, including the paracolic gutter and rectovesical space, indicating charcoal-induced granulomas rather than peritoneal carcinomatosis. PMID- 18431144 TI - Pitfalls of I-131 whole body scan interpretation: bronchogenic cyst and mucinous cystadenoma. AB - Whole body iodine scans are routinely performed in the nuclear medicine department as part of the management of differentiated thyroid carcinoma. Similarly, radioactive iodine has a well-established role as an adjunct to thyroidectomy in the treatment of these patients. A thorough understanding of the normal, benign, and pathologic biodistribution of iodine is imperative for the nuclear medicine physician. This knowledge leads to the accurate determination of the presence of metastatic or recurrent carcinoma, and may even facilitate the accurate detection of an undiagnosed condition. Above all, correct image interpretation avoids unnecessary therapeutic doses. The authors describe 2 unusual examples of false positive findings in fluid-filled cavities that showcase the variety of nonmalignant entities one may encounter when interpreting metastatic surveys. PMID- 18431145 TI - Detection of medullary thyroid cancer with MIBG imaging for pheochromocytoma. AB - A medullary thyroid cancer (MTC)/multiple endocrine neoplasia syndrome was suspected in a patient having an metaiodobenzylguanidine (MIBG) scan while she was being investigated for pheochromocytoma. After surgery, this was confirmed histologically. Although MIBG scanning cannot be used as a screening tool for MTC because of its poor sensitivity for detection of MTC, this case report highlights that one should always scrutinize the thyroid gland while interpreting MIBG scans. PMID- 18431146 TI - Costosternal chondrodynia simulating recurrent breast cancer unveiled by FDG PET. AB - FDG-PET imaging, which is routinely used to assess recurrent breast cancer, is nonspecific for malignancies and can detect several inflammatory lesions. Costochondritis following breast reconstructive surgery, presenting with excruciating chest pain or "chondrodynia," is a rare variant of Tietze's syndrome thought to occur due to traction exerted by large breast implants. Diagnosis is usually clinical because no abnormalities are identified on conventional imaging. Two cases have been reported in the literature, but none with FDG-PET. We report a patient with treated breast cancer, presenting with chest pain where FDG-PET to assess for recurrence was consistent with costochondritis and peri-implant inflammation. PMID- 18431147 TI - SPECT/CT of axillofemoral graft infection. AB - A 73-year-old woman was admitted to the hospital via the emergency room with increasing pain and tenderness in the right axilla and high fevers. She had a long history of peripheral vascular disease with multiple graft failures and recent insertion of a right axillofemoral graft. A computed tomography study of the chest obtained at admission confirmed graft patency and a small fluid collection around the graft at the level below the right axilla. It also confirmed a saccular aneurysm of the aortic arch. A SPECT/CT study of the chest at 3 hours after reinjection of autologous leukocytes labeled with Tc-99m showed intense labeled leukocyte uptake around the vascular graft in the lateral chest wall. Culture of the infected tissue around the excised graft grew Staphylococcus aureus sensitive to all tested antibiotics. Vascular graft infections are rare, with few reported cases of axillofemoral graft infections utilizing Ga-67. Labeled leukocytes have been used successfully in the setting together with CT scanning. This is a rare case of labeled leukocyte uptake in an infected axillofemoral graft by SPECT/CT. PMID- 18431148 TI - Interarterial course of the right coronary artery: assessment with cardiac computed tomography and myocardial perfusion imaging. AB - A 50-year-old well-trained cyclist reported increasing brief episodes of chest pain over the last 8 years at maximum exercise during alpine bicycle training. Previous cardiac stress testing on a supine bicycle ergometer revealed nonspecific ST-T abnormalities. In June 2007, the patient was referred for computed tomography coronary angiography, which revealed a coronary anomaly, with the right coronary artery originating from the left coronary sinus and taking an interarterial course between the ascending aorta and the pulmonary trunk. There were no atherosclerotic alterations in the coronary arteries. Subsequent bicycle exercise stress and myocardial perfusion imaging revealed no myocardial perfusion defect. PMID- 18431149 TI - Peri-aortofemoral prosthesis urinoma: diagnosis by Tc-99m MAG3 SPECT/CT and differentiation from simultaneous hepatobiliary excretion. AB - A 75-year-old woman developed renal failure 1 week after elective aortobifemoral bypass surgery. Postoperative computed tomography showed right hydronephrosis. Tc 99m mercaptoacetyltriglycerine (MAG3) scintigraphy was performed to exclude renal obstruction or acute tubular necrosis. Planar MAG3 images demonstrated right hydronephrosis and unusual accumulation of tracer between the kidneys and the right upper quadrant of abdomen, with new areas of activity in the right lower quadrant on delayed images. SPECT/CT demonstrated MAG3 activity within fluid collections adjacent to the aorta and right iliac/inguinal arteries, consistent with a urine leak. The right upper quadrant activity represented MAG3 accumulation within the gallbladder. PMID- 18431150 TI - Endotracheal metastasis from squamous cell cancer of the head and neck: PET/CT imaging. AB - Lung cancer is the main malignancy with frequent endotracheal and endobronchial dissemination. Extrapulmonary sources of airway metastasis are less common, predominantly seen with breast, colorectal, and renal neoplasms. Head and neck cancer represents only a small fraction of tumors with tracheal luminal metastasis. We present 2 such rare cases with F-18 FDG PET/CT imaging. These 2 companion cases show in one instance an obvious hypermetabolic endotracheal lesion and in the other one a discrete airway metastasis overlooked by chest CT and diagnosed by PET with coregistered CT. PMID- 18431151 TI - Benign tongue FDG uptake in a patient with tardive dyskinesia. AB - Fluorine-18 fluorodeoxyglucose positron emission tomography-computed tomography (F-18 FDG PET-CT) is the modality of choice for diagnosis, staging, and restaging of many malignancies. The importance of eliminating false positives cannot be underestimated because they can dramatically alter the clinical course. We present a case of benign uptake in the tongue secondary to tardive dyskinesia in a 62-year-old woman referred for staging of ductal carcinoma of the breast who was concurrently receiving oral therapy for schizoaffective disorder. This case emphasizes the importance of direct clinical interview and adequate history taking in the formulation of an appropriate diagnosis. PMID- 18431152 TI - Findings of intramediastinal gossypiboma with F-18 FDG PET in a melanoma patient. AB - FDG PET/CT scan was performed to evaluate recurrence in an asymptomatic 64-year old man with a history of melanoma in the left posterior ear. PET/CT images showed an intense ring-shaped area of FDG activity in the posterior mediastinum in a large posterior mediastinal mass. However, further evaluation indicated that this activity was caused by an intramediastinal gossypiboma after coronary artery bypass graft surgery 4 years before the PET/CT scan. PMID- 18431153 TI - Potential false positive Tc-99m sestamibi parathyroid study due to uptake in brown adipose tissue. AB - We report on a 55-year-old woman with suspected primary hyperparathyroidism who underwent dual phase Tc-99m sestamibi parathyroid imaging. Symmetric, patchy activity in the neck and shoulders was localized to low attenuation areas on integrated SPECT/CT and attributed to uptake in brown adipose tissue (BAT). Focal uptake in the anterior thorax on SPECT images, which potentially may have been misinterpreted as ectopic parathyroid tissue, was demonstrated on SPECT/CT as uptake in BAT. Recognition of this pattern on parathyroid SPECT/CT scintigraphy may avoid false positive reports. Our case provides further evidence that in addition to F-18 FDG, I-123 MIBG, and Tc-99m tetrofosmin, Tc-99m sestamibi may also accumulate in BAT. PMID- 18431154 TI - Cold exposure-induced rhabdomyolysis demonstrated by bone scintigraphy. AB - This report describes a case of acute renal failure due to rhabdomyolysis that developed after prolonged exposure to cold. A 47-year-old man visited a mountain during winter wearing only casual clothes and became lost at temperatures below freezing for 8 hours. Upon admission, the patient complained of a tingling sensation and numbness in the both upper and lower extremities with anuria. The serum blood urea nitrogen, creatinine, creatinine kinase, and myoglobin levels were elevated. Bone scintigraphy on the second day revealed increased activity in the superficial muscle groups in the abdomen, back, buttock, thighs, legs, and arms, which was compatible with rhabdomyolysis. PMID- 18431155 TI - FDG uptake in abdominal mesh implant on FDG PET/CT. AB - A 50-year-old man with peritoneal angiosarcoma underwent total tumor excision along with removal of the total right rectus and medial part of the left rectus muscles 1 year earlier. F-18 fluorodeoxyglucose (FDG) positron emission tomography/computed tomography (PET/CT) was performed for suspected recurrence. In the anterior abdominal wall extensive subcutaneous FDG uptake showing linear hypermetabolic zone on PET slices was noted. The patient had a history of a dual mesh implant, which consists of polypropylene and polyvinylidene fluoride for the reinforcement of the abdominal wall during the operation. PET/CT fusion image demonstrated that the finding was due to the dual mesh implant in the anterior abdominal wall most likely representing a foreign body reaction. PMID- 18431156 TI - Desmoplastic neurotropic melanoma in a patient with trigeminal neuralgia: FDG PET/CT and MRI. AB - A 51-year-old man presented with a hard subcutaneous nodule adhering to the underlying bone structures of the left eyelid in 2004. Histopathology showed a desmoplastic neurotropic melanoma (DNM) with perineural invasion. Patient presented with a first recurrence in October 2005, which was treated by surgery. In August 2006 he presented with trigeminal neuralgia of the left face in the area innervated by the first and second branches of the trigeminal nerve. A PET scan clearly shows the tumoral hypermetabolism of the subcutaneous recurrence of the neurotropic melanoma with invasion of the second or maxillary branch that follows the nerve up to the trigeminal ganglion, which was detected despite the physiological high uptake in the temporal lobe. MRI confirms the invasion and a second PET and MRI 6 months later done to evaluate resectability showed progressive disease. DNM is a rare subtype of spindle cell melanoma. It corresponds to dermal proliferation of desmoplastic cells of neural differentiation. Unlike other melanomas, however, survival for DNM may be better compared with other forms of melanoma. This rare case report presents PET imaging involving cranial nerve invasion by this uncommon melanoma subtype. PMID- 18431157 TI - Spinal cord metastases from lung cancer: detection with F-18 FDG PET/CT. AB - Preoperative F-18 FDG PET/CT study in this 57-year-old woman showed an FDG avid lesion in the left upper lung without evidence of lymphadenopathy or distant metastasis. She underwent a left upper lobectomy in June 2005 revealing moderately poorly differentiated adenocarcinoma (pT3N0M0) and subsequent chemotherapy completed December 2005. Nine months later, a left parietal lobe metastatic lesion was surgically resected. A true whole body FDG PET/CT study in November 2006 for restaging demonstrated new FDG avid spinal cord foci, which were highly suspicious for spinal cord metastases; these lesions were confirmed by MRI. Clinically, the patient recently developed back pain without evidence of neurologic deficits. PMID- 18431158 TI - Disseminated tuberculosis masquerading as metastatic breast carcinoma on PET-CT. AB - PET has an established role in the management of breast cancer. However, F-18 FDG uptake sometimes has been associated with benign disease leading to false positive results. We present a case of a 37-year-old woman who presented with a 3 month history of a left breast lump and palpable left axillary lymph nodes. Whole body PET-CT scan demonstrated multiple focal areas of intense FDG uptake in the left breast and multiple axillary, cervical, and mediastinal lymph nodes. PET-CT findings mimicked metastatic breast cancer, which was subsequently confirmed as disseminated tuberculosis by mammotome-guided biopsy of the breast lesion and fine needle aspiration biopsy of lymph nodes. PMID- 18431159 TI - Demonstration of a perihepatic abscess communicating with a fistula from the small bowel on hepatobiliary imaging. AB - Abscess formation after abdominal surgery is not an uncommon complication. It is much less common for a collection to be the result of a fistulous tract from the bowel. We describe a patient who underwent a Tc-99m hepatobiliary (Choletec) scan for the workup of a perihepatic abscess, which confirmed the presence of a fistulous tract from the small bowel to a perihepatic collection. PMID- 18431160 TI - Disseminated extranodal NK/T-cell lymphoma, nasal type, with multiple subcutaneous nodules: utility of 18F-FDG PET in staging. AB - A 49-year-old man presented with a left nasal cavity mass, biopsy of which revealed extranodal NK/T cell lymphoma. A PET-CT scan showed increased F-18 FDG uptake in the nasal mass, anterior mediastinal lymph nodes, and multiple subcutaneous nodular deposits in the chest wall, gluteal region, and right femoral areas. The patient achieved complete remission with salvage l asparaginase therapy after failing first-line standard anthracycline-based chemotherapy. PET-CT was very useful in detecting subcutaneous nodules consistent with widespread dissemination. There is limited literature data on the use of the PET scan in the diagnosis and staging of extranodal nasal NK/T-cell lymphomas. Our case highlights the important role of F-18 FDG PET in the staging of these patients. PMID- 18431161 TI - Detection of multiple myeloma by PET/CT in a patient with colon cancer. AB - A 76-year-old man with previous sigmoid colon resection for adenocarcinoma had low back pain for 2 months. Whole-body bone scintigraphy showed multiple focal Tc 99m methylene diphosphonate (MDP)-avid lesions in both rib cages and 3 lumbar vertebrae, indicating metastases. F-18 fluorodeoxyglucose (FDG) positron emission tomography (PET)/computed tomography (CT) imaging was performed for further evaluation of this possible metastatic disease and demonstrated the lumbar and costal metastases and several hypermetabolic areas in the pelvic bones, multiple thoracic vertebrae, both shoulders, and the right femur. Histopathologic examination of the right-sided iliac crest, however, revealed multiple myeloma. PMID- 18431162 TI - Stress fracture of the first rib in a weightlifter. AB - A 20-year-old woman elite weightlifter presented with acute onset debilitating right shoulder pain while attempting the "clean and jerk." There was no previous relevant history, including no prior fracture in the region. Examination confirmed vague tenderness in the right supraclavicular fossa with no discernible neurologic or vascular deficit within the right arm. X-ray was normal. The patient had a bone scan, including SPECT, suggesting a fracture of the right first rib, subsequently confirmed on CT examination. PMID- 18431163 TI - Tc-99m red blood cell imaging in a patient with blue rubber bleb nevus syndrome. AB - Blue rubber bleb nevus syndrome, first discovered in 1860 by Gascoyen, is a rare syndrome characterized by multiple venous malformations (hemangiomas) of the skin and in the gastrointestinal tract. These hemangiomas may be found in the skin, GI tract, central nervous system, thyroid, parotid, eyes, oral cavity, musculoskeletal system, lungs, kidneys, liver, spleen, and bladder, and carry a significant potential for serious bleeding. We report a case of a 9-year-old patient who presented with bloody stools and was referred for localization of the bleeding site. A Tc-99m labeled red blood cell (RBC) scan showed no evidence of active lower GI bleeding but demonstrated multiple venous malformations throughout the body. Tc-99m labeled red blood cell imaging may be helpful to delineate the extent of involvement in patients with blue rubber bleb nevus syndrome. PMID- 18431165 TI - Current readings in nuclear medicine. PMID- 18431164 TI - Hashimoto thyroiditis and Takayasu aortitis: visualization of the thyroid gland and ring appearance of the mediastinum on F-18-FDG PET. AB - A woman with Hashimoto thyroiditis, treated with levothyroxine for 4 years, complained of chest pain that radiated to the neck and had progressively worsened for 2 months. CT of the chest showed circumferential thickening of the ascending aorta and aortic arch, compatible with Takayasu arteritis. She underwent F-18 FDG PET imaging, which revealed an abnormal thyroid gland and a ring-like appearance of the mediastinum. PMID- 18431174 TI - Administering a transdermal drug. PMID- 18431175 TI - Caps off to this sewing project. PMID- 18431176 TI - What is tumor lysis syndrome? AB - A patient in our hematology unit who was receiving chemotherapy for lymphoma developed a complication called tumor lysis syndrome. Can you explain what this is?-M.K., GA. PMID- 18431188 TI - About dysgraphia. PMID- 18431197 TI - Dealing with an angry patient. AB - Most nurses have been verbally or physically assaulted. Use these tips to defuse a potentially dangerous situation. PMID- 18431198 TI - Seizures. PMID- 18431199 TI - How to recognize and respond to refeeding syndrome. AB - If your patient is malnourished, you need to assess her carefully before she restarts nutrition by any route to avoid potentially lethal consequences. PMID- 18431202 TI - Seeking refuge from the storm. AB - I didn't think I had much to offer my home health care patient, but I unexpectedly met some of her needs that snowy day. PMID- 18431203 TI - Responding to a fire emergency. AB - Practicing the right moves will help you respond quickly and confidently if smoke or a fire threatens. PMID- 18431204 TI - Extreme heart makeover: understanding mechanical circulatory support. AB - Learn about ventricular assist devices and total artificial hearts and how they affect patient care. PMID- 18431206 TI - When your patient has atopic dermatitis. PMID- 18431210 TI - Bracing and splinting to manage pain. PMID- 18431212 TI - Comparing short peripheral cannula insertion sites. PMID- 18431214 TI - Start early to prevent genital HPV infection-and cervical cancer. PMID- 18431215 TI - Tension pneumothorax. PMID- 18431216 TI - Clinical diagnosis of ventriculoperitoneal shunt failure among children with hydrocephalus. AB - OBJECTIVE: To define the significance of various symptoms and signs in the diagnosis of ventriculoperitoneal shunt failure and infection. METHODS: The observations that form the basis of this study were made in the course of 2 multicenter, prospective, randomized, controlled clinical trials of technical aspects of ventriculoperitoneal shunt surgery-the Shunt Design Trial and the Endoscopic Shunt Insertion Trial. At registration, basic demographic and baseline clinical data were recorded. At scheduled follow-up visits 3 months and 1, 2, and 3 years after surgery and at unscheduled visits, the presence or absence of various symptoms or signs was recorded. At each visit, the neurosurgeon investigator made a determination about whether the shunt had reached an end point: mechanical obstruction, infection, overdrainage, or loculation of the ventricular system. Observations at the last follow-up visit for each patient constituted the data for the current study. Sensitivities, specificities, and likelihood ratios were calculated for each symptom and sign as tests for shunt failure from any cause and for failure by infection. Decision trees were constructed to analyze the relationships of various symptoms and signs in the diagnosis of shunt failure and infection. RESULTS: Observations were available for analysis from 647 patient visits. A total of 248 shunts were judged to have failed (38%), and 55 were judged specifically to have failed by infection (8.5%). Bulging fontanel, fluid collection along the shunt, depressed level of consciousness, irritability, abdominal pain, nausea and vomiting, abnormal shunt pump test, accelerated head growth, and headache were strongly associated with shunt failure. Fever was strongly associated with shunt infection. Gross signs of wound infection were associated with shunt infection but were observed infrequently. Decision tree analysis confirmed the salience of bulging fontanel as a predictor of shunt failure. Fever and time since initial surgery were powerful predictors of shunt infection. Irritability emerged as an important observation in the identification of both shunt failure and shunt infection. Among children who underwent initial shunt insertion after 2 months of age, the absence of irritability, nausea/vomiting, and headache were powerful and generalizable predictors of the absence of shunt failure or infection. CONCLUSIONS: Analysis of symptoms and signs of ventriculoperitoneal shunt complications can inform clinical judgment in the assessment of children with hydrocephalus. PMID- 18431217 TI - Current utilization of continuous end-tidal carbon dioxide monitoring in pediatric emergency departments. AB - OBJECTIVE: End-tidal carbon dioxide (ETCO2) monitoring has numerous clinical applications in the emergency setting. This study was designed to explore the current availability and utilization patterns for continuous ETCO2 monitoring in pediatric emergency departments. METHODS: A Web-based survey was distributed to directors of all accredited pediatric emergency medicine fellowship programs in the United States and Canada. RESULTS: Eighty-one percent of directors completed this survey. Eighty-eight percent had access to ETCO2 monitoring for intubated patients and 53% for nonintubated patients. Seventy-nine percent of respondents used ETCO2 monitoring "always" or "often" for endotracheal tube confirmation. Only 20% of respondents used ETCO2 monitoring "always" or "often" for moderate sedation, 16% for trauma, and 6% for acid-base disturbances. One hundred percent of respondents who used ETCO2 monitoring felt that it was easy to use. The most common reason for not using ETCO2 monitoring was lack of equipment (65%). CONCLUSIONS: ETCO2 monitoring is widely available, yet underutilized, for spontaneously breathing patients in pediatric emergency departments. PMID- 18431218 TI - Acceptability of tobacco cessation interventions in the pediatric emergency department. AB - OBJECTIVES: To assess the acceptability to parents and staff of providing a brief tobacco cessation intervention incorporating the first 2 A's of the Clinical Practice Guideline on Treating Tobacco Use and Dependence (Ask and Advise) + fax referral to a state tobacco quitline from the emergency department (ED) of a large children's hospital. METHODS: Data were collected from a written survey of (1) parental smokers accompanying their children to a pediatric ED who consented and were randomized to participate in a tobacco cessation intervention and (2) medical doctors (MDs) or nurse practitioners (NPs) caring for their child. Acceptability of the intervention was measured on a 5-point Likert scale ranging from "strongly agree" to "strongly disagree." RESULTS: Of the 1643 parental subjects that were screened to participate in this study, 557 (33.9%) were smokers, 240 of whom were enrolled in the intervention group and a total of 215 (90%) completed the survey. A total of 230 (100%) of the MDs/NPs approached completed the survey. Of the 215 parental subjects in the intervention group, 212 (98.6%) and 208 (96.7%) agreed that the intervention provided them with "useful information" and "easy to understand" advice, respectively. Most parental subjects agreed that the advice given was sufficient and met their needs (n = 208, 96.7%) and that the study length was "just right" (n = 206, 95.8%). On a rating scale of 1 to 10, the mean (SD) rating of the study was 9.1 (1.3). Of the MDs/NPs participating in this survey, 224 (97.4%) and 206 (89.6%) agreed that the "ED is a good place to screen parents for tobacco use" and that the "ED is a good place to give advice about tobacco cessation," respectively. In addition, 202 (87.8%) and 196 (85.2%) reported that they felt comfortable giving tobacco cessation advice to parents or referring parents to the national quitline, respectively. Only 7 (3%) felt that the study interfered with patient care. CONCLUSIONS: An ED tobacco cessation intervention using the 2 A's + Quitline referral had excellent acceptability in this study of parents and staff of pediatric patients. The use of the pediatric ED as a venue to providing tobacco cessation counseling to a population with a high prevalence of parental smokers warrants further consideration. PMID- 18431219 TI - Nasal foreign bodies in children: should they have a plain radiograph in the accident and emergency? AB - OBJECTIVES: PRIMARY OBJECTIVE: To determine the proportion of children with nasal foreign bodies who had button batteries as the foreign body. SECONDARY OBJECTIVE: To describe the clinical care and outcome of those children with a button battery foreign body. METHODS: A retrospective review of all children presenting to the accident and emergency room with a nasal foreign body or unilateral nasal discharge during a 6-month period was assessed. RESULTS: Forty four children were included. The most common object found was a plastic bead (27%), followed by foam, paper, or tissue fragments (23%); food matter represented 15%. A button battery was found in 3 patients (7%); other foreign bodies included stones, buttons, crayons, erasers, and a pellet. Thiry-six (82%) had their foreign body removed without anesthetic and 8 (18%) required a general anesthetic. All 3 children with a button battery went undiagnosed until examination in the operating theater. Because there was no previous indication of the presence of a button battery in the 3 children, the removal of the foreign body under general anesthesia was scheduled for the following day. CONCLUSIONS: The removal of the button batteries would have been expedited had a plain radiograph been obtained, possibly resulting in less morbidity for our patients. We recommend a plain radiograph in all children presenting with a nonvisible foreign body or unilateral nasal discharge. PMID- 18431220 TI - Observational study of helmet use among children skiing and snowboarding. AB - OBJECTIVES: The objectives of this study were to observe the use of helmets among school-age children while skiing and snowboarding and to determine factors that influenced helmet use in children. METHODS: This was a prospective observational study. Children estimated to be school age (5-17 years of age) were observed at a Western New York area ski resort as they entered ski areas. Data collected included estimated age, helmet use, sex, sport, and whether the children were accompanied by adults or other children or were alone. RESULTS: During February 2005, a total of 1472 children were observed. Thirty-seven percent wore a helmet. Helmets were worn by 42% of skiers and 32% of snowboarders. Sixty percent of children wore helmets when observed with an adult, compared with 28% when observed with other children. Those children on beginner slopes (47%) were more likely to wear helmets than those on intermediate slopes (34%). There was no difference in helmet use among males and females. CONCLUSIONS: Despite recommendations for children to wear helmets while participating in winter sports, about a third of the children observed wore a helmet. Children skiing, younger children, and children on beginner slopes were more likely to wear helmets. Future studies are needed to determine if the use of helmets would decrease the incidence of head injuries while skiing and snowboarding. PMID- 18431221 TI - A statement from the transplantation society against organ trafficking and commerce in organs. PMID- 18431222 TI - Searching the transplantation library. PMID- 18431223 TI - Airway transplantation: a debate worth having? AB - Laryngeal and tracheal transplantation have been proposed as treatments for irreversible airway disease for many years. Despite much research, there has only been one true laryngeal transplant reported. Although this was in many ways a success, several barriers remain before full clinical trials. There are issues over patient selection, reinnervation, immunosuppression, and cost-benefit. For the trachea, where finely tuned neuromuscular activity is not an issue, tissue engineering probably represents the future. This overview discusses the arguments for and against laryngeal and tracheal transplantation and suggests ways of overcoming these barriers. PMID- 18431224 TI - How much donor human leukocyte antigen-specific antibody is too much for a renal transplant? PMID- 18431225 TI - Ischemia and cholestasis: more than (just) the bile ducts! PMID- 18431226 TI - Clinical relevance of pretransplant donor-directed antibodies detected by single antigen beads in highly sensitized renal transplant patients. AB - BACKGROUND: Highly sensitized (HS) patients (>85% panel-reactive antibodies) have a lower chance of receiving a donor kidney. Within Eurotransplant the Acceptable Mismatch (AM) program was developed to increase the chances of HS patients to receive a crossmatch-negative donor kidney. The standard crossmatch in the AM program is based on complement-dependent cytotoxicity. METHODS: In this study we wanted to determine the clinical relevance of human leukocyte antigen donor directed antibodies (DDA) detected by the single antigen (SA) bead technique, in the pretransplant sera of HS patients transplanted in our center through the Eurotransplant AM program. RESULTS: From 34 AM patients, 27 were transplanted with 1 to 5 mismatches and 7 received a 0-mismatched graft. From the mismatched patients, retrospectively, 13 proved to possess pretransplant DDA by SA whereas 14 did not. No antibodies were found in the 0-mismatched group. Comparison of the DDA+ and DDA- patients in the human leukocyte antigen-mismatched donor/recipient combinations revealed a trend to an earlier and higher number of rejection episodes in DDA+ patients (P=0.08). No detrimental effect of DDA on graft survival was observed. CONCLUSIONS: This single-center study showed that in the AM program DDA detected by SA, and not by less-sensitive methods, may be related to acute rejection episodes but is not detrimental to long-term graft outcome. These findings question the increasing use of more-sensitive screening techniques for the allocation of organs. PMID- 18431227 TI - Effects of glucose toxicity and islet purity on in vivo magnetic resonance imaging of transplanted pancreatic islets. AB - BACKGROUND: Pancreatic islet transplantation has recently emerged as a powerful clinical modality to restore normoglycemia in diabetic patients. Despite the success of the Edmonton protocol, these patients still experience a significant islet loss immediately after transplantation. Noninvasive magnetic resonance imaging (MRI) allows for longitudinal monitoring of graft loss providing that islets are labeled with a magnetically "visible" contrast agent. To fully interpret the imaging data, it is critical to investigate factors normally present during clinical transplantation and influencing MRI of transplanted islets. METHODS: Here, we focused on both the effect of hyperglycemia and the effect of contaminating nonendocrine tissue, which is always present in islet preparations, on MRI imaging of islet grafts. Human pancreatic islets labeled with Feridex were transplanted in diabetic and healthy animals. Separate groups of animals were transplanted with Feridex-labeled pure and nonpure (50% islets and 50% nonendocrine tissue) preparations. The fate of the graft in all groups was monitored by in vivo MRI. RESULTS: We found that diabetic animals with transplanted islets showed a significantly higher rate of islet death than their healthy counterparts on in vivo MR images. Interestingly, transplantation of islets contaminated with nonendocrine tissue did not have any significant influence on MR images, presumably because of a low labeling rate of this tissue and a fast rate of its disappearance after transplantation. CONCLUSIONS: We believe that this study serves as yet another step on our way to clinical use of in vivo imaging of islet transplantation. PMID- 18431228 TI - Does reduction in immunosuppression in viremic patients prevent BK virus nephropathy in de novo renal transplant recipients? A prospective study. AB - BACKGROUND: BK virus nephropathy (BKVN) is a severe complication of renal transplantation, resulting in graft loss in >50% of cases. Because patients with BKV viremia are at high risk for developing BKVN, the aim of our study was to analyze whether early reduction of immunosuppression (IS) could prevent BKVN in viremic patients. METHODS: BKV viruria was prospectively screened every 3 months by real-time polymerase chain reaction during the first year after transplantation in 123 consecutive renal transplant recipients. Plasma viral load was measured by polymerase chain reaction whenever viruria was positive; in viremic patients a graft biopsy was systematically performed and IS was reduced. RESULTS: Viruria, viremia, and BKVN occurred in 48.8%, 10.5%, and 2.4% of patients, respectively. In the 13 patients with positive viremia, initial graft biopsy showed BKVN in two. After reduction of IS in patients without BKVN, viremia disappeared in 8 of 11, decreased in 2 of 11, and increased in one patient who eventually developed BKVN. In contrast, viremia remained positive in one patient with BKVN and disappeared in the second but renal function deteriorated in both of them. Initial viral load was higher in patients who developed BKVN. CONCLUSION: Reduction of IS is probably an effective therapeutic option to clear viremia and prevent BKVN in viremic renal transplant patients. PMID- 18431229 TI - Viral persistence after liver transplantation for hepatitis B virus: a cross sectional study. AB - BACKGROUND: Prophylaxis to prevent recurrent HBV infection in liver transplant (LT) recipients has evolved over time, and we manage patients who receive lamivudine monoprophylaxis, lamivudine with HBV immunoglobulin (HBIg), and lamivudine and adefovir with HBIg. METHODS: Serum was examined with sensitive assays to detect the persistence of HBV, and to identify mutations that might confer resistance to the antiviral prophylaxis. Forty patients were studied, and sera were collected 20 days to 13.3 years after LT. RESULTS: Overall, HBV DNA was detected in serum of 67.5% of patients (8 of 10 of lamivudine monoprophylaxis patients, 15 of 24 of those receiving lamivudine and HBIg, and 4 of 6 of those receiving lamivudine, adefovir and HBIg). Thus, HBV infection persists for most of the patients despite successful prophylaxis after LT. Of those patients with detectable serum HBV DNA, three of eight of the lamivudine monoprophylaxis group had sequences associated with resistance to lamivudine (YMDD mutants), compared with only 1 of 15 of the lamivudine and HBIg cohort. Three of the lamivudine and HBIg cohort had the I126A Hepatitis B surface antigen escape variant. In those serum HBV DNA-positive patients who were receiving lamivudine, adefovir, and HBIg, only one of four had YMDD mutant, and none had Hepatitis B surface antigen escape variants. None of the 40 patients suffered clinical HBV recurrence. CONCLUSIONS: Our observations imply that the selection of resistant virus may be essential, but is not sufficient to cause overt failure of prophylaxis with development of clinical disease. It seems likely that the patients' immune response contributes, at least partially, to the long-term control of infection in these patients. PMID- 18431230 TI - Increase of proliferating renal progenitor cells in acute tubular necrosis underlying delayed graft function. AB - BACKGROUND: Delayed graft function (DGF) is associated with acute tubular necrosis. In this setting, surviving tubular cells may proliferate and replace injured cells. CD133Pax-2cells may play a role in the regeneration of tubular damage. The aim of this study was to demonstrate the presence of these cells in human kidneys before transplantation and in grafts with DGF. METHODS: Ten normal kidneys (group 1) and pretransplant biopsy of 25 deceased donors (group 2) were examined. The latter group included 10 kidneys with early graft function (2A) and 15 with DGF (2B). Group 2B patients received a second biopsy during DGF (2C). CD133, Pax-2, and Ki-67 protein expression was investigated by confocal microscopy. RESULTS: CD133Pax-2 and CD133Pax-2cells were present within the Bowman's capsule and proximal tubules in all groups except group 2B. Number of CD133Pax-2 and CD133Pax-2cells at tubular level was similar in groups 1 and 2A. Within group 2B we observed a striking reduction in both cell types. There was a significant increase of both cell populations within group 2C, compared with group 2B. CD133Pax-2 and CD133Pax-2cell number in group 2 correlated inversely with cold ischemia time. Pax-2Ki-67cells were absent from group 1 and 2B samples, and increased significantly in groups 2A and 2C. Proliferating CD133 cells increased significantly in group 2C. CONCLUSIONS: Our data suggest that regenerative response in posttransplant acute tubular necrosis, underlying DGF, is characterized by an increase in proliferating renal progenitor/stem cells CD133Pax-2 and CD133Pax-2 cells involved in repairing tubular damage. PMID- 18431231 TI - Negative impact of one-year anemia on long-term patient and graft survival in kidney transplant patients receiving calcineurin inhibitors and mycophenolate mofetil. AB - BACKGROUND: The impact of posttransplant anemia (PTA) upon patient and graft survival remains controversial. The aim of this study was to assess the incidence of PTA 1 year after transplantation in patients treated with calcineurin inhibitors and mycophenolate mofetil, and to determine the impact of 1-year PTA upon long-term patient and graft survival. METHODS: Between January 1, 1999, and December 31, 2003, all patients with a functioning graft at 1 year, and who were receiving an immunosuppressive regimen based on calcineurin inhibitors and mycophenolate mofetil, were included in the study (n=339). Anemia was defined according to the World Health Organization criteria, that is, hemoglobin levels less than 13 g/dL for men and less than 12 g/dL for women. RESULTS: One hundred and eight of 339 were anemic at 1 year after transplantation (31.85%; group I). Independent predictors for 1-year anemia are donor's age and serum creatinine at 6 months. At last follow-up, that is, 69.4+/-17.7 months after transplantation, there had been a significant number of deaths in group I (n=7; 6.9%) compared with nonanemic patients (group II) (n=4; 1.73%; P=0.04). Mean allograft survival was significantly better in group II (70.7+/-17.1 months) compared with group I (66.4+/-18.7 months; P=0.03). Also, 12 graft losses (11.1%) were observed in group I and seven occurred in group II (3%; P=0.004). Independent predictors for allograft loss included delayed graft function and serum creatinine at 1 year. CONCLUSION: After kidney transplantation, the occurrence of PTA at 1 year is harmful, in the long term, to patient survival. PMID- 18431232 TI - Alemtuzumab preconditioning with tacrolimus monotherapy-the impact of serial monitoring for donor-specific antibody. AB - BACKGROUND: Antibody preconditioning with tacrolimus monotherapy has allowed many renal allograft recipients to be maintained on spaced weaning. METHODS: Of 279 renal allograft recipients transplanted between March 2003 and December 2004, 222 (80%) had spaced weaning (i.e., reduction of tacrolimus monotherapy dosing to every other day, three times a week, twice a week, or once a week) attempted. Routine monitoring for donor-specific antibody (DSA) was begun in September 2004. Mean follow-up is 34+/-6.5 months after transplantation and 26+/-8.1 months after the initiation of spaced weaning. RESULTS: One hundred and twenty-two (44%) patients remained on spaced weaning. One- and 2-year actual patient/graft survival was 99%/99%, and 97%/96%. Fifty-six (20%) patients experienced acute rejection after initiation of spaced weaning. One- and 2-year actual patient/graft survival was 100%/98%, and 94%/78%. Forty-two (15%) patients with stable renal function had spaced weaning stopped because of the development of DSA, which disappeared in 17 (40%). One- and 2-year actual patient and graft survival was 100% and 100%. CONCLUSION: Adult renal transplant recipients who are able to be maintained on spaced weaning have excellent outcomes. Patients with stable renal function who have reversal of weaning because of the development of DSA also have excellent outcomes. Routine monitoring for DSA may allow patients to avoid late rejection after spaced weaning. PMID- 18431233 TI - Prediabetes in patients receiving tacrolimus in the first year after kidney transplantation: a prospective and multicenter study. AB - BACKGROUND: Tacrolimus-based immunosuppression, the most widely used regimen in kidney transplantation, increases the risk of new onset diabetes after transplantation (NODAT). However, the prevalence, evolution and risk factors of different prediabetic alterations: impaired fasting glucose, impaired glucose tolerance, and provisional diabetes, have not been established. METHODS: In this multicenter and prospective study we evaluated 154 nondiabetic kidney transplant recipients receiving tacrolimus, mycophenolate mofetil and low dose steroids. An oral glucose tolerance test was performed 3 and 12 months after transplantation and prediabetes was defined by American Diabetes Association criteria. RESULTS: Prediabetes was highly prevalent and showed little variation between 3 and 12 months (36% and 33%, respectively). Impaired glucose tolerance was the most frequent abnormality observed (23% and 25%, respectively) observed. In addition, 20% of recipients showed NODAT by 1 year. Multivariate analysis showed that age (odds ratio [OR]: 1.07, 95% confidence interval [CI]: 1.004-1.14), pretransplant body mass index (OR: 1.3, CI: 1.09-1.6) and triglyceride/high density lipoprotein cholesterol ratio, a marker of insulin resistance, (OR: 1.4, CI: 1.05-1.9) were independent risk factors for prediabetes. CONCLUSION: One in two recipients with tacrolimus-based immunosuppresion showed prediabetes or NODAT by 1 year posttransplantation when properly investigated. Older age and high pretransplant body mass index and triglyceride/high density lipoprotein-cholesterol ratio were risk factors for prediabetes. These findings may help applying early interventions to prevent the disorder. PMID- 18431234 TI - Maturation of dose-corrected tacrolimus predose trough levels in pediatric kidney allograft recipients. AB - BACKGROUND: In contrast to adult kidney recipients, in whom the long-term evolution and clinical determinants of tacrolimus pharmacokinetics are well studied, less is known about the long-term evolution of tacrolimus pharmacokinetics in pediatric kidney transplant recipients. METHODS: One-hundred and five pediatric recipients of a kidney allograft, all treated with a corticosteroid-free immunosuppressive protocol, were included. The evolution of tacrolimus doses and predose trough (C0) levels was recorded at 3, 6, 9, 12, 18, and 24 months after transplantation, as well as all C0 levels obtained in the first 2 years after transplantation. The evolution and clinical determinants of tacrolimus exposure parameters were analyzed. RESULTS: Dose-corrected tacrolimus C0 levels (C0/dose/kg) increased in the first 2 years after kidney transplantation in pediatric recipients (P=0.001). This decrease in dose requirement by time was only significant in children older than 5 years at the time of transplantation (P=0.38, 0.03, and 0.001 for age groups <5, 5-12, and >12 years, respectively). In addition, the younger patients had significantly higher dose requirements (dose/kg) compared with older recipients (P=0.0002). CONCLUSION: Pediatric kidney transplant recipients exhibit maturation of dose corrected tacrolimus predose trough levels with time after transplantation. This cannot be explained by differences in corticosteroid use, because all patients were treated with a corticosteroid-free protocol. The higher dose requirements for younger recipients and the absence of tacrolimus maturation in the youngest recipients suggest that age-dependent changes in tacrolimus intestinal first-pass effect, metabolism, or distribution play a role. Whether age-specific tacrolimus dosing algorithms will improve outcome needs further study. PMID- 18431235 TI - Long-term effect of folic acid therapy in heart transplant recipients: follow-up analysis of a randomized study. AB - BACKGROUND: Folic acid therapy reduces homocysteine plasma levels, which seem to influence occurrence of cardiac allograft vasculopathy, but its effect on medium- or long-term prognosis after heart transplantation is unknown. METHODS: We analyzed 7-year outcome of 51 recipients randomized to receive 15 mg/day of methyltertrahydrofolate for 1 year after heart transplantation or standard therapy alone (originally, for intravascular ultrasound study of short-term cardiac allograft vasculopathy progression); recipients were observed for a further 5 to 6 years. RESULTS: Overall, 13 deaths occurred (six oncologic, five cardiovascular, two infective). Estimated 7-year survival was better in recipients randomized to folate (88%+/-6% vs. 61%+/-9%, P=0.04). After adjusting for age, pretransplant coronary artery disease, and hyperhomocysteinemia, posttransplant folic acid therapy was associated with lower mortality (relative risk [RR] 0.53, 95% confidence interval [CI] 0.25-0.97; P=0.036), apparently driven by reductions in both cancer-related and cardiovascular causes. Reduced mortality was marked in a high-risk subgroup comprising older recipients and patients transplanted because of coronary artery disease (RR 0.43, 95% CI 0.17 0.85) but not in the lower-risk subgroup (RR 1.11, 95% CI 0.22-5.61). CONCLUSIONS: Although further studies are needed, it seems reasonable to suggest folate therapy to heart transplant recipients. It is possible that properties other than homocysteine reduction may provide antitumoral benefits. PMID- 18431236 TI - Analysis of tolerance induction using triple chimeric mice: major histocompatibility complex-disparate thymus, hemopoietic cells, and microenvironment. AB - BACKGROUND: Although bone marrow transplantation (BMT) has become a valuable strategy for the treatment of various intractable diseases in recent years, success rates remain low in elderly patients because of low thymic function. We have previously shown that fetal thymus transplantation (TT) with BMT is effective for elderly recipients in mice. METHODS: We performed fully major histocompatibility complex (MHC)-mismatched fetal TT from B6 (H-2) mice plus allogeneic BMT from C3H/HeN (H-2) mice by intra-bone marrow-BMT (IBM-BMT) using congenitally athymic nude (nu/nu) BALB/c (H-2), or BALB/c adult-thymectomized recipients to obtain triple chimeras. We next carried out the IBM-BMT+TT using senescence-accelerated mouse P1 strain (SAMP1) to examine whether this method would be applicable to aging mice. RESULTS: Triple chimeric mice survived for a long period with sufficient T-cell functions comparable to the mice treated with BMT plus MHC-matched TT, whereas those without TT survived for a short period with insufficient T-cell reconstitution. Almost all the hematolymphoid cells were derived from donor bone marrow cells. Interestingly, they showed tolerance to all three types of MHC determinants with donor-derived thymic dendritic cells in TT. Triple chimeric SAMP1 also survived for long periods with T-cell functions restored in contrast to non-TT SAMP1 recipients. CONCLUSION: These findings suggest that third party combined TT with allogeneic IBM-BMT may be more advantageous for elderly recipients with low thymic function, than IBM-BMT alone (without TT). PMID- 18431237 TI - Dearterialization of the liver causes intrahepatic cholestasis due to reduced bile transporter expression. AB - BACKGROUND: Bile duct injury after hepatic artery thrombosis (HAT) in liver transplantation is believed to be caused by ischemia predominantly. We aimed to define the involvement of bile secretory dysfunction in the pathogenesis of liver injury after HAT. METHODS: In a murine model, the main hepatic artery, the extrahepatic peribiliary plexus, or both arterial connections to the liver were interrupted (n=5 for each group). After 1, 14, or 28 days, hepatobiliary function was assessed by analysis of bile transporter expression, serum bile acids and bilirubin, and hepatic ATP content. In addition, cellular injury was assessed by light microscopy and biochemical markers. RESULTS: There were no signs of hepatobiliary dysfunction or injury in sham-operated animals or in mice with interruption of the hepatic artery or the extrahepatic peribiliary plexus alone. However, as early as 24 hr after complete dearterialization, bile transporter expression was significantly reduced and intrahepatic cholestasis started to progress the following weeks. Histologic studies at 28 days after complete dearterialization showed severe hepatobiliary injury. CONCLUSIONS: This study indicates that arterial blood supply is critical for normal bile secretion. Bile duct injury after complete arterial deprivation is preceded by a loss of bile secretory function and subsequent intrahepatic cholestasis. PMID- 18431238 TI - Depleting anti-CD4 monoclonal antibody (GK1.5) treatment: influence on regulatory CD4+CD25+Foxp3+ T cells in mice. AB - BACKGROUND: CD4(+)CD25(+) regulatory T (Treg) cells are often essential for the maintenance of immunologic self-tolerance and transplant tolerance in some cases. The effects of depleting anti-CD4 monoclonal antibody (GK1.5), which was used in transplant tolerance induction, on CD4(+)CD25(+) Treg cells have not been investigated. METHODS: Three weeks after BALB/c mice were injected with GK1.5 or phosphate-buffered saline, the levels, phenotype and immunosuppressive function of CD4(+)CD25(+) Treg cells in these mice were detected. RESULTS: The numbers of CD4 and CD4(+)CD25(+) Treg cells in the periphery were markedly decreased in GK1.5-treated mice. However, GK1.5 treatment significantly enhanced the ratios of CD4(+)CD25(+) T cells or CD4(+)CD25(+)Foxp3 T cells to CD4(+) T cells in the periphery (P<0.01). Compared with the control mice, more CD4(+)CD25(+) T cells in GK1.5-treated mice showed CD45RB and CD62L phenotype. Furthermore, enriched CD4(+)CD25(+) Treg cells in GK1.5-treated mice show immunosuppressive ability on the immune response of T effector cells to alloantigens or mitogen as efficiently as those from the control mice in vitro. CONCLUSIONS: GK1.5 could significantly enhance the percentage of CD4(+)CD25(+)Foxp3(+) Treg cells in the periphery while keeping these cells functional, indicating that GK1.5 might affect the potential induction of immune tolerance by different influences on CD4(+)CD25(+)Treg cells and CD4(+)CD25(-) T cells in periphery. PMID- 18431239 TI - Fms-related tyrosine kinase 3 expression discriminates hematopoietic stem cells subpopulations with differing engraftment-potential: identifying the most potent combination. AB - BACKGROUND: Fms-related tyrosine kinase 3 (Flt3)-ligand (FL) promotes the proliferation, differentiation, development, and mobilization of hematopoietic cells. We previously found that FL-mobilized hematopoietic stem cells (HSC) engraft efficiently, whereas FL-expanded bone marrow HSC do not. The function of FL-mobilized c-Kit(+) Sca-1(+)Lin(-)(KSL) subpopulations has not been systematically evaluated. A precise definition of the repopulating ability is needed to define which HSC subpopulations are critical for long-term chimerism and tolerance induction. FL significantly mobilized c-Kit(hi) and c-Kit(lo) Sca 1(+)Lin(-) cells into peripheral blood (PB). Here, we evaluated the influence of Flt3 expression on long-term repopulating ability of HSC subpopulations. METHODS: c-Kit(hi) or c-Kit(lo) KSL cells were sorted from PB of FL-treated green fluorescent protein-positive donors. The function of these cells was evaluated using competitive reconstitution assays, colony-forming units spleen, and colony forming cell assays. The function of c-Kit(hi) CD34(-)Flt3(-) KSL, c-Kit CD34(+)Flt3(-) KSL, c-Kit(hi) CD34(+)Flt3(+) KSL were investigated in an in vivo transplantation model. RESULTS: Only FL-mobilized PB c-Kit(hi) KSL cells exhibited high spleen colony-forming unit activity, generated high numbers of both lymphoid and myeloid colonies in vitro, and rescued ablated recipients. FL mobilization expanded both c-Kit(hi) CD34(+)Flt3(-) cells (short-term HSC) and c Kit(hi) CD34(-)Flt3(-) KSL cells (long-term HSC). There was a significant decrease in c-Kit CD34Flt3 KSL late multipotent progenitors in PB. A combination of c-Kit(hi) CD34Flt3 and c-Kit CD34(+)Flt3(-) KSL cells offered the most effective rescue of ablated recipients. CONCLUSIONS: These data suggest that engraftment of purified HSC is influenced by both short- and long-term repopulating populations and that Flt3 expression may be useful for selecting the most critical HSC subpopulations for transplantation. PMID- 18431240 TI - Characterization of naturally occurring CD4+CD25+ regulatory T cells in rhesus monkeys. AB - BACKGROUND: Translational research in a relevant preclinical model is recommended before Treg-inducing protocols can be implemented in humans. We have characterized rhesus monkey CD25 cells phenotypically and functionally. METHODS: The phenotype of CD4(+)CD25(high) cells was determined by FACS, focusing on established markers of mouse and human Treg cells. Percentages of cells positive for CD45RA, CD62L, and intracellular CTLA-4 and FOXP3 were compared between CD4(+)CD25(high) and CD4(+)CD25(-) cells. CD25 cells stimulated with anti-CD3, ConA, and/or allogeneic peripheral blood mononuclear cells were mixed with freshly isolated CD25 cells. The suppressive activity of the CD25 cells in vitro was assessed using several experimental conditions. RESULTS: Rhesus monkey CD4(+)CD25(high) cells expressed high intracellular levels of CTLA-4 and FOXP3, whereas expression was negligible in CD4(+)CD25(-) cells. The CD25(high) population was mostly CD45RA(-), indicative of a memory phenotype. The CD25(+) cells were anergic, because they showed low proliferative responses, no interleukin-2 production, and some interferon-gamma and interleukin-10 production. Proliferation of CD4(+)CD25(-) cells stimulated by anti-CD3 or allogeneic cells was decreased when CD4(+)CD25(-) cells were added at a 1:1 ratio. In addition, we found that CD25 cells inhibited the interleukin-2 and interferon-gamma production by anti-CD3-stimulated CD25 cells in a dose-dependent fashion, through a cell-cell contact-dependent mechanism. CONCLUSIONS: Rhesus monkey CD4(+)CD25(+) cells have similar phenotypic and functional characteristics as natural Tregs in humans. These findings allow testing of Treg expansion and induction protocols in a relevant preclinical model, the rhesus monkey. PMID- 18431242 TI - Pretransplant donor-specific antibodies in cytotoxic negative crossmatch kidney transplants: are they relevant? AB - BACKGROUND: The corresponding antigens of alloantibodies identified in patients awaiting kidney transplantation are often listed as unacceptable for transplantation. The use of solid phase testing, being more sensitive and accurate than conventional complement-dependent cytotoxicity (CDC) assays, has resulted in increased identification of alloantibodies. We aimed to study the clinical importance of alloantibodies defined solely by solid phase techniques. METHODS: All patients transplanted between 1999 and 2001 at our center with available day-of-transplant sera (D0) were included (121 patients). All had negative CDC crossmatches. RESULTS: Thirty-eight patients (31%) had detectable alloantibodies using high-definition assays with 16 having donor-specific antibodies (DSA) and 22 non-DSA. There were no cases of hyperacute rejection in any of the groups. Biopsy-proven acute rejection rates in the DSA and non-DSA were similar to the unsensitized group. Delayed graft function and 1-year graft survival rates were also similar for the three groups as were median 1-year serum creatinine levels. Multivariate analysis, however, showed that DSA were associated with an increased relative risk of longer-term graft failure (relative risk, 6.5; P<0.05). CONCLUSIONS: These data show that in the context of a CDC negative crossmatch, the presence of D0 DSA has little impact on any early graft parameters. DSA, however, are associated with poorer longer-term graft outcomes in kidney transplantation. PMID- 18431241 TI - Acute antibody-mediated complement activation mediates lysis of pancreatic islets cells and may cause tissue loss in clinical islet transplantation. AB - BACKGROUND: Clinical islet transplantation is associated with loss of transplanted islets necessitating tissue from more than one donor to obtain insulin independence. The instant blood-mediated inflammatory reaction (IBMIR) is one explanation to the tissue loss. Complement activation is an important cytotoxic component of the IBMIR, and in the present study, we have investigated this component in detail. METHODS: Isolated human islets were analyzed by large particle flow cytometry and confocal microscopy after incubation in human ABO compatible hirudin-plasma. RESULTS: After incubation in plasma, the islets bound IgG and IgM, CIq, C4, C3 and C9. The binding of C3b/iC3b was evident already after 5 min. The binding of C3b/iC3b and the generation of C3a and sC5b-9 were inhibited by the complement inhibitor Compstatin. Lysis as reflected by propidium iodide (PI) staining and release of C-peptide was also inhibited by Compstatin. There were significant correlations between IgM/IgG versus C3b/iC3b and between sC5b-9 and C-peptide. CONCLUSION: The conclusion is that complement is activated by natural IgG and IgM antibodies already after 5 min. The complement activation leads to lysis of cells of the pancreatic islets. This very rapid reaction may be an essential entity of the damage induced by the IBMIR in clinical islet transplantation. PMID- 18431243 TI - CD4+ T cells are sufficient to elicit allograft rejection and major histocompatibility complex class I molecule is required to induce recurrent autoimmune diabetes after pancreas transplantation in mice. AB - BACKGROUND: We characterized the role of T cell subsets and major histocompatibility complex molecules in allograft rejection and recurrence of autoimmune diabetes. METHODS: Adoptive cell transfer and vascularized segmental pancreas transplantation were performed in mice. RESULTS: In an alloimmune response model, transfer of nondiabetic CD4, but not CD8 T cells, elicited pancreas allograft rejection in streptozotocin (STZ)-induced diabetic NOD/scid mice. Pancreas allografts were acutely rejected in STZ-induced diabetic NOD/beta2m mice (confirmed the absence of major histocompatibility complex [MHC] class I and CD8 T cells) and permanently accepted in NOD/CIIT mice (confirmed the absence of MHC class II and CD4 T cells). The results suggest that rejection of pancreas allograft is CD4-dependent and MHC class I-independent. In the autoimmune diabetes model, whole spleen cells obtained from diabetic NOD mice induced autoimmune diabetes in NOD/scid and NOD/CIIT mice, but the onset of diabetes was delayed in NOD/beta2m mice. However, the purified diabetic T cells failed to elicit autoimmune diabetes in NOD/beta2m mice. NOD/scid and NOD/CIIT pancreas grafts were acutely destroyed whereas four of six NOD/beta2m pancreas grafts were permanently accepted in autoimmune diabetic NOD mice. CONCLUSION: CD4 T cells are sufficient for the induction of allograft rejection, and MHC class I molecule is required to induce recurrent autoimmune diabetes after pancreas transplantation in mice. PMID- 18431244 TI - Kidney transplant histology after one year of continuous therapy with sirolimus compared with tacrolimus. AB - Analysis of 1-year surveillance biopsies was carried out for kidney transplant recipients participating in a randomized trial comparing tacrolimus- and sirolimus-based immunosuppression. The analysis was restricted to recipients remaining on assigned regimen throughout the first posttransplant year. Biopsies from 57 of 84 (68%) tacrolimus-randomized recipients were compared with 38 of 81 (47%) of sirolimus-randomized recipients, the discrepancy being explained by a higher rate of sirolimus discontinuation for non-graft-related complications. Included recipients from the two groups did not differ for baseline characteristics or 1-year iothalamate clearance. Histologic analysis indicated no differences between the groups for glomerular, arterial/arteriolar, or acute interstitial abnormalities. There were, however, significantly higher mean scores in the tacrolimus group for interstitial fibrosis and tubular atrophy with a trend toward higher estimated percent interstitial fibrosis. The results indicate that sirolimus may be associated with reduced early graft fibrosis compared with tacrolimus. This potential benefit is offset by lower success rate in maintaining the regimen and was not accompanied by superior glomerular filtration rate at 1 year. PMID- 18431245 TI - Incidence, predictors, and outcomes of cardiac pacing after cardiac transplantation: an 11-year retrospective analysis. AB - More than 20% of cardiac transplant patients go on to require permanent pacing. We sought to determine the incidence of cardiac pacing in our cardiac transplant population and identify characteristics that may predict which patients will require permanent pacing. We reviewed medical records of cardiac transplant recipients and compared baseline characteristics of patients who received pacemakers with those of patients who did not receive pacemakers. Of 292 patients included in this analysis, 71 (24%) required permanent posttransplant pacing. Use of amiodarone before transplant was associated with a nonsignificant trend toward needing a pacemaker after transplant (P=0.08). Patients undergoing biatrial anastomosis were more likely to require permanent pacing than patients undergoing bicaval anastomosis (P<0.001). Approximately one fourth of cardiac transplant patients require permanent pacing. Surgical technique is a major predictor of who will require permanent pacing after cardiac transplantation. PMID- 18431246 TI - Posttransplant lymphoproliferative disorder followed by Hodgkins disease in a renal transplant recipient. PMID- 18431247 TI - Occult hepatitis B is not necessarily an infection. PMID- 18431248 TI - A case of valerian-associated hepatotoxicity. PMID- 18431249 TI - A new procedure for 16 consecutive days monitoring of intragastric pH. PMID- 18431250 TI - Accumulation of aspartic acid421- and glutamic acid391-cleaved tau in neurofibrillary tangles correlates with progression in Alzheimer disease. AB - Truncations of tau protein at aspartic acid421 (D421) and glutamic acid391 (E391) residues are associated with neurofibrillary tangles (NFTs) in the brains of Alzheimer disease (AD) patients. Using immunohistochemistry with antibodies to D421- and E391-truncated tau (Tau-C3 and MN423, respectively), we correlated the presence of NFTs composed of these truncated tau proteins with clinical and neuropathologic parameters in 17 AD and 23 non-AD control brains. The densities of NFTs composed of D421- or E391-truncated tau correlated with clinical dementia index and Braak staging in AD. Glutamic acid391 tau truncation was prominent in the entorhinal cortex, whereas D421 truncation was prominent in the subiculum, suggesting that NFTs composed of either D421- or E391-truncated tau may be formed mutually exclusively in these areas. Both truncations were associated with the prevalence of the apolipoprotein E epsilon4 allele. By double labeling, intact tau in NFTs was commonly associated with D421-cleaved tau but not with E391 truncated tau; D421-cleaved tau was never associated with E391-truncated tau. These results indicate that tau is not randomly proteolyzed at different domains, and that proteolysis occurs sequentially from the C-terminus to inner regions of tau in AD progression. Identification of NFTs composed of tau at different stages of truncation may facilitate assessment of neurofibrillary pathology in AD. PMID- 18431251 TI - Knocking down galectin 1 in human hs683 glioblastoma cells impairs both angiogenesis and endoplasmic reticulum stress responses. AB - Galectin (Gal) 1 is a hypoxia-regulated proangiogenic factor that also directly participates in glioblastoma cell migration. To determine how Gal-1 exerts its proangiogenic effects, we investigated Gal-1 signaling in the human Hs683 glioblastoma cell line. Galectin 1 signals through the endoplasmic reticulum transmembrane kinase/ribonuclease inositol-requiring 1alpha, which regulates the expression of oxygen-regulated protein 150. Oxygen-regulated protein 150 controls vascular endothelial growth factor maturation. Galectin 1 also modulates the expression of 7 other hypoxia-related genes (i.e. CTGF, ATF3, PPP1R15A, HSPA5, TRA1, and CYR61) that are implicated in angiogenesis. Decreasing Gal-1 expression in Hs683 orthotopic xenografts in mouse brains by siRNA administration impaired endoplasmic reticulum stress and enhanced the therapeutic benefits of the proautophagic drug temozolomide. These results suggest that decreasing Gal-1 expression (e.g. through brain delivery of nonviral infusions of anti-Gal-1 siRNA in patients) can represent an additional therapeutic strategy for glioblastoma. PMID- 18431252 TI - Activation of the heat shock response in familial amyloidotic polyneuropathy. AB - The heat shock proteins (Hsps) have been implicated in a variety of neurodegenerative diseases in which the underlying pathology is protein aggregation. Here, we studied the heat shock response in familial amyloidotic polyneuropathy (FAP), a neurodegenerative disease caused by aggregation and extracellular tissue deposition of mutated transthyretin (TTR). We observed greater expression of Hsp27 and Hsp70 related to the presence of extracellular TTR aggregates in human FAP nerve, skin, and salivary gland biopsies than in normal controls. Transthyretin aggregates did not colocalize with Hsp, suggesting that extracellular TTR tissue deposits induce an intracellular stress response. Moreover, the heat shock transcription factor 1 was upregulated and localized to nuclei in affected tissues. Transgenic mice expressing the V30M mutant form of TTR similarly showed the presence of TTR deposits, induced activation of heat shock transcription factor 1, and increased synthesis of Hsp. Furthermore, the addition of toxic TTR aggregates to cultures of human and rodent neuroblastoma cell lines induced upregulation of Hsp70 and Hsp27. Taken together, these novel findings suggest new avenues for research on pathogenic mechanisms in FAP and identify the heat shock response as a potential pharmacologic treatment target for FAP. PMID- 18431253 TI - Endothelial cell barrier impairment induced by glioblastomas and transforming growth factor beta2 involves matrix metalloproteinases and tight junction proteins. AB - Gliomas, particularly glioblastoma multiforme, perturb the blood-brain barrier and cause brain edema that contributes to morbidity and mortality. The mechanisms underlying this vasogenic edema are poorly understood. We examined the effects of cocultured primary cultured human glioblastoma cells and glioma-derived growth factors on the endothelial cell tight junction proteins claudin 1, claudin 5, occludin, and zonula occludens 1 of brain-derived microvascular endothelial cells and a human umbilical vein endothelial cell line. Cocultured glioblastoma cells and glioma-derived factors (e.g. transforming growth factor beta2) enhanced the paracellular flux of endothelial cell monolayers in conjunction with downregulation of the tight junction proteins. Neutralizing anti-transforming growth factor beta2 antibodies partially restored the barrier properties in this in vitro blood-brain barrier model. The involvement of endothelial cell-derived matrix metalloproteinases (MMPs) was demonstrated by quantitative reverse transcriptase-polymerase chain reaction analysis and by the determination of MMP activities via zymography and fluorometry in the presence or absence of the MMP inhibitor GM6001. Occludin, claudin 1, and claudin 5 were expressed in microvascular endothelial cells in nonneoplastic brain samples but were significantly reduced in anaplastic astrocytoma and glioblastoma samples. Taken together, these in vitro and in vivo results indicate that glioma-derived factors may induce MMPs and downregulate endothelial tight junction protein and, thus, play a key role in glioma-induced impairment of the blood-brain barrier. PMID- 18431254 TI - Coenzyme q induces tau aggregation, tau filaments, and Hirano bodies. AB - Tau aggregation is a common feature of tauopathies such as Alzheimer disease (AD). In AD, tau assembles into fibrillar polymers; it may also be present in other aberrant aggregates, including Hirano bodies. The mechanisms leading to tau polymerization in vivo are not understood. In this study, we found that coenzyme Q (ubiquinone) facilitates tau aggregation after binding to tau molecules at the region of the tau molecule involved in self-assembly. Consequently, after tau-tau interactions, this region is masked in fibrillar tau polymers. Further in vitro studies showed that ubiquinone facilitates the interaction of tau protein with actin to form structures that are morphologically similar to Hirano bodies. Finally, studies in AD brains show that Hirano bodies react with an antibody raised against ubiquinone, indicating that ubiquinone is a component of Hirano bodies. Taken together, the in vitro models and findings in AD suggest that in the presence of ubiquinone, Hirano bodies may result from the interaction of actin and other proteins, including tau. PMID- 18431255 TI - Trauma-induced cell swelling in cultured astrocytes. AB - Brain edema and associated increased intracranial pressure are major consequences of traumatic brain injury that account for most early deaths after traumatic brain injury. An important component of brain edema after traumatic brain injury is astrocyte swelling (cytotoxic edema). To examine the pathophysiologic mechanisms of trauma-induced astrocyte swelling, we used an in vitro fluid percussion trauma model. Exposure of cultured rat astrocytes to 5 atm of pressure resulted in significant cell swelling at 1 to 24 hours posttrauma that was maximal at 3 hours. Because oxidative/nitrosative stress, mitochondrial permeability transition (mPT), and mitogen-activated protein kinases (MAPKs) have been implicated in astrocyte swelling in other neurologic conditions, we examined their potential roles in this model. We previously showed increased free radical generation after in vitro trauma and show here that trauma to astrocytes increased the production of nitric oxide. Trauma also induced mPT and increased phosphorylation (activation) of MAPKs (extracellular signal-regulated kinase 1/2, c-Jun-N-terminal kinase, and p38-MAPK); these changes were diminished by antioxidants and the nitric oxide synthase inhibitor N-nitro-l-arginine methyl ester. Antioxidants, N-nitro-l-arginine methyl ester, the mPT inhibitor cyclosporin A, and inhibitors of MAPKs all significantly diminished trauma induced astrocyte swelling. These findings demonstrate that direct mechanical injury to cultured astrocytes brings about cell swelling, and that blockade of oxidative/nitrosative stress, mPT, and MAPKs significantly reduce such swelling. PMID- 18431257 TI - Plasmacytoid dendritic cells in multiple sclerosis: intracerebral recruitment and impaired maturation in response to interferon-beta. AB - The roles of plasmacytoid dendritic cells (pDCs) and their response to interferon (IFN)-beta therapy in multiple sclerosis (MS) patients are poorly understood. We identified pDC accumulation in white matter lesions and leptomeninges of MS brains and abundant expression of the Type I IFN-induced protein MxA, mainly in perivascular CD3+ lymphocytes in lesions, indicating Type I IFN production by activated pDCs. The pDC chemoattractant chemerin was detected in intralesional cerebrovascular endothelial cells, and the chemerin receptor was expressed on infiltrating leukocytes, including pDCs. The effect of IFN-beta on pDC phenotype and function was evaluated in MS patients before and during IFN-beta treatment. Although IFN-beta did not modify the frequency and immature phenotype of circulating pDC, they showed lower expression of major histocompatibility complex Class II and blood-dendritic cell antigen 2 molecules and upregulation of CD38 and B7H1 costimulatory molecules. On exposure to CpG (a site where cytosine [C] lies next to guanine [G] in the DNA sequence [the p indicates that C and G are connected by a phosphodiester bond]) oligodeoxynucleotides in vitro, pDCs from IFN-beta-treated MS patients showed reduced expression of the pDC maturation markers CD83 and CD86 molecules; in vitro IFN-beta treatment of pDCs from healthy donors resulted in lower secretion of proinflammatory cytokines, including IFN alpha, and a decreased ability to stimulate allogeneic T cells in response to maturative stimuli. These data indicate that IFN-beta modulates the immunologic functions of pDC, thus identifying pDCs as a novel target of IFN-beta therapy in MS patients. PMID- 18431259 TI - Effect of empowerment on professional practice environments, work satisfaction, and patient care quality: further testing the Nursing Worklife Model. AB - The purpose of this study was to test Leiter and Laschinger's Nursing Worklife Model linking structural empowerment to Lake's 5-factor professional practice work environment model and work quality outcomes. A predictive, nonexperimental design was used to test the model in a random sample of 234 staff nurses. The analysis revealed that professional practice environment characteristics mediated the relationship between structurally empowering work conditions and both job satisfaction and nurse-assessed patient care quality. PMID- 18431260 TI - The Stability and Workload Index for Transfer score predicts unplanned intensive care unit patient readmission: initial development and validation. AB - OBJECTIVE: Unplanned readmission of hospitalized patients to an intensive care unit (ICU) is associated with a worse outcome, but our ability to identify who is likely to deteriorate after ICU dismissal is limited. The objective of this study is to develop and validate a numerical index, named the Stability and Workload Index for Transfer, to predict ICU readmission. DESIGN: In this prospective cohort study, risk factors for ICU readmission were identified from a broad range of patients' admission and discharge characteristics, specific ICU interventions, and in-patient workload measurements. The prediction score was validated in two independent ICUs. SETTING: One medical and one mixed medical-surgical ICU in two tertiary centers. PATIENTS: Consecutive patients requiring >24 hrs of ICU care. INTERVENTIONS: None. MEASUREMENTS: Unplanned ICU readmission or unexpected death following ICU dismissal. RESULTS: In a derivation cohort of 1,131 medical ICU patients, 100 patients had unplanned readmissions, and five died unexpectedly in the hospital following ICU discharge. Predictors of readmission/unexpected death identified in a logistic regression analysis were ICU admission source, ICU length of stay, and day of discharge neurologic (Glasgow Coma Scale) and respiratory (hypoxemia, hypercapnia, or nursing requirements for complex respiratory care) impairment. The Stability and Workload Index for Transfer score predicted readmission more precisely (area under the curve [AUC], 0.75; 95% confidence interval [CI], 0.70-0.80) than the day of discharge Acute Physiology and Chronic Health Evaluation III score (AUC, 0.62; 95% CI, 0.56-0.68). In the two validation cohorts, the Stability and Workload Index for Transfer score predicted readmission similarly in a North American medical ICU (AUC, 0.74; 95% CI, 0.67-0.80) and a European medical-surgical ICU (AUC, 0.70; 95% CI, 0.64 0.76), but was less well calibrated in the medical-surgical ICU. CONCLUSION: The Stability and Workload Index for Transfer score is derived from information readily available at the time of ICU dismissal and acceptably predicts ICU readmission. It is not known if discharge decisions based on this prediction score will decrease the number of ICU readmissions and/or improve outcome. PMID- 18431258 TI - DNA damage and repair: relevance to mechanisms of neurodegeneration. AB - DNA damage is a form of cell stress and injury that has been implicated in the pathogenesis of many neurologic disorders, including amyotrophic lateral sclerosis, Alzheimer disease, Down syndrome, Parkinson disease, cerebral ischemia, and head trauma. However, most data reveal only associations, and the role for DNA damage in direct mechanisms of neurodegeneration is vague with respect to being a definitive upstream cause of neuron cell death, rather than a consequence of the degeneration. Although neurons seem inclined to develop DNA damage during oxidative stress, most of the existing work on DNA damage and repair mechanisms has been done in the context of cancer biology using cycling nonneuronal cells but not nondividing (i.e. postmitotic) neurons. Nevertheless, the identification of mutations in genes that encode proteins that function in DNA repair and DNA damage response in human hereditary DNA repair deficiency syndromes and ataxic disorders is establishing a mechanistic precedent that clearly links DNA damage and DNA repair abnormalities with progressive neurodegeneration. This review summarizes DNA damage and repair mechanisms and their potential relevance to the evolution of degeneration in postmitotic neurons. PMID- 18431261 TI - Serum thrombomodulin level relates to the clinical course of disseminated intravascular coagulation, multiorgan dysfunction syndrome, and mortality in patients with sepsis. AB - OBJECTIVE: To determine serum concentrations of thrombomodulin, the marker of endothelial injury, in patients with sepsis-induced disseminated intravascular coagulation and multiple organ dysfunction syndrome and to investigate the independent association between this marker and the development of disseminated intravascular coagulation, multiple organ dysfunction syndrome, and mortality. DESIGN: A prospective cohort study. SETTING: A 37-bed intensive care unit of a tertiary care hospital. PATIENTS: One hundred consecutive patients with sepsis. INTERVENTIONS: Serum thrombomodulin concentrations and the development of disseminated intravascular coagulation and multiple organ dysfunction syndrome were determined in patients on days 1 and 3 of sepsis. These data were used to determine an association between day 1 thrombomodulin concentrations and development of disseminated intravascular coagulation, multiple organ dysfunction syndrome, and mortality during intensive care unit stay. These connections were determined by the Cox proportional hazards model and plotting of receiver operating characteristic curves. MEASUREMENTS AND MAIN RESULTS: Day 1 serum concentrations of thrombomodulin were higher in patients with disseminated intravascular coagulation (11.1 +/- 1.0 vs. 5.3 +/- 0.5 ng/mL; p < .0001) or multiple organ dysfunction syndrome (10.3 +/- 0.7 vs. 4.3 +/- 0.4 ng/mL; p < .0001) than those without, respectively. In patients with resolved disseminated intravascular coagulation (4.9 +/- 0.5 vs. 8.9 +/- 0.9 ng/mL, day 3 vs. day 1, p = .005) or multiple organ dysfunction syndrome (6.3 +/- 1.4 vs. 12.0 +/- 1.6 ng/mL, day 3 vs. day 1, p < .0001) on day 3 of sepsis, day 3 levels of thrombomodulin were down from day 1. Thrombomodulin concentration independently predicted the development of disseminated intravascular coagulation (hazard ratio 1.13, p < .0001), multiple organ dysfunction syndrome (hazard ratio 1.12, p < .0001), and mortality (hazard ratio 1.19, p < .0001) during intensive care unit stay. The area under the receiver operating characteristic curve showed that day 1 serum thrombomodulin levels had good discriminative power in predicting the development of disseminated intravascular coagulation (0.811), multiple organ dysfunction syndrome (0.896), and mortality (0.803) during intensive care unit stay. CONCLUSIONS: Endothelial cell injury is critical in the progression from disseminated intravascular coagulation to multiple organ dysfunction syndrome and subsequent mortality in septic patients. Serum concentrations of thrombomodulin may be used in monitoring disseminated intravascular coagulation and multiple organ dysfunction syndrome in these patients. PMID- 18431262 TI - Temporal changes in management and outcome of septic shock in patients with malignancies in the intensive care unit. AB - OBJECTIVE: Septic shock is a severe, often terminal, complication of malignancy. For patients without malignancy, outcome from septic shock has improved with new advances in care. We wished to explore whether outcome from septic shock has similarly improved for cancer patients, with regard to implementation of recent adjuvant therapies. DESIGN: An 8-yr retrospective observational study. SETTING: A 24-bed medical intensive care unit in a university hospital. PATIENTS: Patients were 238 consecutive cancer patients (solid tumors or hematologic malignancies) with septic shock admitted to the intensive care unit within two consecutive 4-yr periods: 1998-2001 and 2002-2005. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Septic shock occurred in 90 patients in 1998-2001 and 148 in 2002-2005. Management of septic shock between the two periods mostly differed by emergence of adjuvant therapies of sepsis (mainly low-dose glucocorticoids) and intensive insulin therapy and a more frequent use of renal replacement therapy in the recent period. Short-term survival rates were significantly higher during 2002 2005 compared with the previous 4-yr period: 28-day, intensive care unit, and hospital survival rates were 47.3% vs. 27.8% (p = .003), 41.2% vs. 26.7% (p = .02), and 36.5% vs. 21.1% (p = .01), respectively. After adjustment, intensive care unit admission between 2002 and 2005 was an independent favorable prognostic factor for short-term outcome. Improved survival was mainly observed in patients who did not require renal replacement therapy during their stay in the intensive care unit (hospital survival 65% in 2002-2005 vs. 21.4% in 1998-2001, p < .001). CONCLUSIONS: Improved outcome in critically ill cancer patients extended to the subgroup of patients with septic shock. This might be ascribed both to a better selection of patients and to improvements in the care and management, including new therapeutic strategies for sepsis. PMID- 18431264 TI - Effects of human atrial natriuretic peptide on renal function in patients undergoing abdominal aortic aneurysm repair. AB - OBJECTIVE: Ischemia-reperfusion injury is an important cause of renal dysfunction after abdominal aortic aneurysm repair. Human atrial natriuretic peptide (hANP) is a potent endogenous natriuretic, diuretic, and vasorelaxant peptide. The objective of the present study was to evaluate the effects of hANP on renal function in patients undergoing abdominal aortic aneurysm repair. DESIGN: A prospective, randomized, placebo-controlled study SETTING: Intensive care unit of a university hospital. PATIENTS: Forty patients undergoing elective abdominal aortic aneurysm repair. INTERVENTIONS: The patients were randomized to receive a continuous infusion of either hANP (0.01-0.05 microg/kg/min) (n = 20) or placebo (n = 20) immediately before aortic cross-clamping. The infusion of hANP or placebo continued for 48 hrs. MEASUREMENTS AND MAIN RESULTS: Blood and urine samples were taken before surgery, at admission to the intensive care unit, and on days 1, 2, and 3 postoperatively, for measurement of serum concentrations of sodium, creatinine, and blood urea nitrogen and plasma concentrations of ANP and brain natriuretic peptide (BNP). Urine volume and urinary concentrations of N acetyl-beta-D-glucosaminidase (NAG), sodium, and creatinine were also measured. The mean plasma concentration of ANP was significantly higher in the hANP group than in the placebo group. The mean plasma BNP concentration was significantly lower in the hANP group than in the placebo group. The mean serum concentrations of creatinine and blood urea nitrogen were significantly (p < .05) lower in the hANP group than in the placebo group. The mean urine volume and mean creatinine clearance were significantly (p < .05) higher in the hANP group than in the placebo group. The mean urinary NAG/creatinine ratio was significantly (p < .05) lower in the hANP group than in the placebo group. CONCLUSIONS: The intraoperative and postoperative infusion of low-dose hANP preserved renal function in patients undergoing abdominal aortic aneurysm repair. Further studies are needed to assess the efficacy of prophylactic hANP infusion on perioperative renal outcome. PMID- 18431265 TI - Isolated and reversible impairment of ventricular relaxation in patients with septic shock. AB - OBJECTIVE: Many patients with septic shock and increased cardiac troponin I (cTnI) do not exhibit significant left ventricular systolic dysfunction. We hypothesized that an isolated and reversible impairment of ventricular relaxation may be associated with the increase in cTnI. DESIGN: Prospective, observational study. SETTING: Surgical intensive care unit in a university hospital. PATIENTS: Total of 54 patients with septic shock. INTERVENTIONS: Fractional area change, early diastolic velocity of mitral annulus, flow propagation velocity of early diastolic mitral inflow, cTnI, tumor necrosis factor-alpha, interleukin (IL)-6, 1beta, -8, and -10 were measured at days 1, 2, 3, 4, 7, and 10 after onset of septic shock. Patients were classified into three groups: normal cTnI (group 1), increased cTnI and fractional area change <50% (group 2), and increased cTnI and fractional area change >50% (group 3). MEASUREMENTS AND MAIN RESULTS: A total of 22 patients had an increase in cTnI, 11 with both systolic and diastolic dysfunctions and 11 with isolated impairment of left ventricular relaxation. At day 1, early diastolic velocity of mitral annulus and flow propagation velocity of early diastolic mitral inflow were significantly lower and tumor necrosis factor-alpha, IL-8, and IL-10 significantly higher in groups 2 and 3 compared with group 1. With resolution of septic shock, early diastolic velocity of mitral annulus and flow propagation velocity of early diastolic mitral inflow measured in patients of groups 2 and 3 returned progressively to values observed in group 1, with a parallel normalization of tumor necrosis factor-alpha, IL-8, and IL-10. CONCLUSIONS: Isolated and reversible impairment of left ventricular relaxation, associated with transient increases in cTnI, tumor necrosis factor-alpha, IL-8, and IL-10, was observed in 20% of patients with septic shock. PMID- 18431266 TI - Short- and long-term follow-up of intensive care unit patients after sedation with isoflurane and midazolam--a pilot study. AB - OBJECTIVE: To compare memories from the intensive care unit (ICU) and short- and long-term psychological morbidity in patients after sedation with intravenous midazolam or inhaled isoflurane. DESIGN: Prospective long-term follow-up after randomized controlled trial. SETTING: General ICU at Karolinska University Hospital, Solna, Stockholm. PATIENTS: Forty patients in need of sedation during ventilator treatment. INTERVENTIONS: Patients were randomized to receive isoflurane or midazolam for goal-directed sedation until extubation or for a maximum of 96 hrs. MEASUREMENTS AND MAIN RESULTS: For short-term follow-up, doctors', nurses', and physiotherapists' notes from the 4 days following exposure to the study drugs were reviewed for words indicating adequate or pathologic cognitive and psychological recovery. For long-term follow-up, all 6-month survivors received questionnaires including the ICU Memory Tool (ICU-MT), Hospital Anxiety and Depression Scale (HADS), Impact of Event Scale (IES), and Well-Being Index. Additionally, several screening questions for previous posttraumatic stress symptoms were included. In the short term follow-up, no significant differences were found between groups. In the long-term follow-up, a trend toward fewer hallucinations/delusions after isoflurane sedation than after midazolam (two of ten isoflurane patients vs. five of seven midazolam patients) was found (p = .06). None of the five solely isoflurane-sedated patients reported hallucinations/delusions from the ICU. There was no difference in groups in long term psychological morbidity as measured with HADS and IES. Memories of negative feelings in the ICU (ICU-MT) were associated with high HADS and IES scores (Fisher's exact test, p = .02 and p = .01, respectively). CONCLUSIONS: Sedation of ICU patients with isoflurane may result in fewer delusional memories or hallucinations from the ICU compared with more commonly used intravenous sedation. Memories of negative feelings from the ICU were associated with symptoms of depression or anxiety or symptoms indicating posttraumatic stress disorder. Further study of memory and cognitive/psychological recovery after prolonged isoflurane sedation beyond 96 hrs is warranted. PMID- 18431267 TI - Risk factors associated with the isolation of colistin-resistant gram-negative bacteria: a matched case-control study. AB - OBJECTIVE: The emergence of multidrug-resistant gram-negative bacteria has led to the re-use of colistin, but resistance to this agent has already been reported. We aimed to investigate the potential risk factors for the isolation of colistin resistant Klebsiella pneumoniae, Acinetobacter baumannii, and Pseudomonas aeruginosa from hospitalized patients. DESIGN: Matched case-control study. SETTING: Tertiary care hospital in Athens, Greece. PATIENTS: Case patients were those who had provided a clinical specimen from which a colistin-resistant K. pneumoniae, A. baumannii, or P. aeruginosa was isolated. Controls were selected from a pool of patients who had susceptible to colistin isolates and were matched (1:1) to cases for species of microorganism and site of isolation. Susceptibility to colistin was determined with the Etest. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Data regarding patient demographics, comorbidities, admission to the intensive care unit, prior antibiotic use, and invasive procedures performed were analyzed as risk factors in a matched bivariable model. Variables significantly associated with colistin-resistant isolates (p < .05) were entered in a backward multivariable logistic regression model. Forty-one colistin resistant unique patient isolates were identified from January 1, 2006, until March 31, 2007. These isolates represented infection in 35 of 41 patients. Risk factors significantly associated with the isolation of colistin-resistant isolates were age, duration of intensive care unit stay, [corrected] surgical procedures, use of colistin, use of monobactams, duration of use of colistin and duration of use of antifungal agents [corrected] In the multivariable model, use of colistin was identified as the only independent risk factor (adjusted odds ratio = 7.78, p = .002). CONCLUSIONS: Colistin-resistant K. pneumoniae, A. baumannii, and P. aeruginosa pathogens may be encountered in clinical practice, in association with inappropriate colistin use. To prevent this phenomenon, colistin should be used judiciously, given that treatment options for colistin resistant gram-negative bacteria are limited. PMID- 18431268 TI - Outcomes of patients considered for, but not admitted to, the intensive care unit. AB - OBJECTIVE: The purpose of this study is to evaluate factors associated with decisions to reject patients from medical intensive care unit (MICU) admission and assess the outcome of these patients. DESIGN: Prospective, observational cohort study. SETTING: Large tertiary referral, teaching hospital. PATIENTS: Consecutive patients evaluated for MICU admission but not admitted. MEASUREMENTS: Patient characteristics and demographics, location of evaluation, clinical and laboratory data, major organ system dysfunction, 48-hr patient status, and 6 month mortality. MAIN RESULTS: A total of 1,302 patients were admitted to the MICU, 353 patients were evaluated for the MICU but were not admitted, and 324 patients were used in analysis. Mean age was 68.6 +/- 17.1 yrs, and 57.7% were women. Hospice care was instituted during or immediately after evaluation in 8.3% (n = 27) of cases. MICU care was declined by the patient in 5.2% (n = 17) of evaluations. The mean Acute Physiology and Chronic Health Evaluation (APACHE) II score was 17.4 +/- 6.0. Factors associated with death at 6 months included age, APACHE II score, entering hospice, and patient choice to decline care. Of the patients considered too well to benefit, 9% were admitted to the MICU within 48 hrs and 35.5% died within 6 months; however, no deaths occurred within 48 hrs. CONCLUSIONS: Patients who are considered for critical care are at very high risk of mortality within 6 months. Given that no deaths occurred within 48 hrs and that only 9% needed intensive care unit admission within 48 hrs, the house staff's decision process is safe at this one institution. PMID- 18431269 TI - Noisy pressure support ventilation: a pilot study on a new assisted ventilation mode in experimental lung injury. AB - OBJECTIVE: To describe and evaluate the effects of the new noisy pressure support ventilation (noisy PSV) on lung physiologic variables. DESIGN: Crossover design with four modes of mechanical ventilation. SETTING: Experimental research facility of a university hospital. SUBJECTS: A total of 12 pigs weighing 25.0 36.5 kg. INTERVENTIONS: Animals were anesthetized, the trachea was intubated, and lungs were ventilated with a mechanical ventilator (volume-controlled mode). Acute lung injury was then induced by surfactant depletion. Biphasic intermittent airway pressure/airway pressure release ventilation (BIPAP/APRV) was initiated, and anesthesia depth was decreased to allow spontaneous breathing. After that, each animal was ventilated with four different modes of assisted mechanical ventilation (1 hr each, Latin squares sequence): 1) PSV, 2) PSV combined with intermittent sighs (PSV + Sighs), 3) BIPAP/APRV + spontaneous breathing, and 4) noisy PSV with random variation of pressure support (normal distribution). The mean level of pressure support was set identical in all PSV forms. MEASUREMENTS AND MAIN RESULTS: We found that noisy PSV increased tidal volume variability compared with PSV and PSV + Sighs (19% vs. 5% and 7%, respectively, p < .05) independently from the inspiratory effort; improved oxygenation and reduced venous admixture but did not affect the amount of nonaerated lung tissue as compared with other assisted ventilation modes; reduced mean airway pressure at comparable minute ventilation; redistributed pulmonary blood flow toward nondependent lung regions similar to other PSV forms, whereas BIPAP/APRV + spontaneous breathing did not; and reduced the inspiratory effort and cardiac output in comparison with BIPAP/APRV + spontaneous breathing. CONCLUSIONS: In the surfactant depletion model of acute lung injury, the new noisy PSV increased the variability of the respiratory pattern and improved oxygenation by a redistribution of perfusion toward the ventilated nondependent lung regions with simultaneous lower mean airway pressure, comparable minute ventilation, and no increase in the inspiratory effort or cardiac output. PMID- 18431270 TI - Endotoxin increases plasma leptin and ghrelin levels in dogs. AB - OBJECTIVE: Evaluations of plasma leptin and ghrelin levels and their relations with circulating levels of proinflammatory mediators, stress hormones, and biochemical markers of hepatorenal injury during experimental endotoxemia in dogs. SETTING: Uludag University. DESIGN: Placebo-controlled animal study. ANIMALS: Adult mongrel dogs (n = 16). INTERVENTIONS: Intravenous injection of endotoxin (1 mg/kg) and blood sample withdrawal before and at 0.5-48 hrs posttreatment. MEASUREMENTS AND MAIN RESULTS: Mean baseline plasma leptin and ghrelin levels were 2.4 +/- 0.1 ng/mL and 867 +/- 58 pg/mL, respectively. Plasma leptin and ghrelin increased significantly by 16% (p < .05) and 72% (p < .001) at 0.5 hr, and they remained elevated by 33-41% (p < .001) and 59-74% (p < .001) at 48 hrs after administration of endotoxin, respectively. There was positive correlation (r = .844; p < .001) between plasma leptin and ghrelin levels in endotoxin-treated dogs. Endotoxemia was associated with several-fold elevations in circulating levels of stress hormones, proinflammatory mediators, and hepatorenal injury markers. Plasma leptin and ghrelin levels in endotoxin-treated dogs were correlated with serum nitric oxide (r = .955 and r = .890; p < .001), procalcitonin (r = .825 and r = .716; p < .001), cortisol (r = .823 and r = .786; p < .001), and hepatorenal injury markers (r = .580 to .745 and r = .393 to .574; p < .05 to .01). CONCLUSIONS: Circulating leptin and ghrelin levels increase during endotoxemia, and these increases are associated with elevated levels of proinflammatory mediators, stress hormones, and serum biochemical markers for hepatorenal dysfunction. PMID- 18431271 TI - Neurocognitive performance after cardiopulmonary resuscitation in pigs. AB - OBJECTIVE: To test the feasibility of a neurocognitive test based on operant conditioning in a porcine model of cardiac arrest and cardiopulmonary resuscitation. Furthermore, to characterize the influence of different durations of cardiac arrest on cognitive performance and the accompanying neurohistopathological changes. DESIGN: Randomized controlled laboratory animal study. SETTING: Animal research facility of a university hospital. SUBJECTS: Seventeen male domestic pigs. INTERVENTIONS: Animals were anesthetized and mechanically ventilated before arterial and pulmonary artery catheters were inserted. Cardiac arrest was induced electrically after randomization of the animals into two groups (n = 7/group) left untreated for either 5 or 8 mins. Cardiopulmonary resuscitation was performed with 100% oxygen and cardiac compressions at 100/min for 5 mins before defibrillation was attempted. Three animals treated identically, with the exception that neither cardiac arrest was induced nor cardiopulmonary resuscitation was performed, served as controls. MEASUREMENTS AND MAIN RESULTS: Hemodynamic variables as well as variables of gas exchange were measured at baseline and 10, 60, 120, 240, and 360 mins after cardiopulmonary resuscitation. Neurocognitive performance was evaluated using a test based on operant conditioning 5 days before and 4 days after cardiopulmonary resuscitation. On the fifth postoperative day, animals were killed and the brains removed for histopathological evaluation of vulnerable brain regions. No noteworthy differences in hemodynamics or gas exchange were observed at baseline or after cardiopulmonary resuscitation. Animals exposed to 8 mins of untreated cardiac arrest showed severe neurocognitive dysfunction, which was statistically significant on postoperative days 2 and 3 in comparison to animals exposed to 5 mins of cardiac arrest or controls. Neurohistopathological evaluation revealed a significantly greater proportion of ischemically damaged neurons in the caudate nucleus and putamen in pigs subjected to 8 mins of cardiac arrest. CONCLUSIONS: Neurocognitive testing is feasible in this setting. Performance worsens with increasing ischemia time and is structurally associated with alterations in the caudate nucleus and the putamen. PMID- 18431272 TI - Catecholamines induce an inflammatory response in human hepatocytes. AB - OBJECTIVE: The liver is an early target organ in sepsis, severe sepsis, and septic shock, contributing to multiple organ failure, and both lipopolysaccharide and gut-derived catecholamines are implicated in the occurrence of hepatocellular dysfunction. Treatment of septic shock involves administration of vasoactive agents such as exogenous catecholamines or vasopressin in order to reestablish blood pressure. As a prelude to clinical application, we tested the hypothesis that catecholamines could modulate the lipopolysaccharide-induced inflammatory response and function in human liver. DESIGN: An in vitro human cell culture study. SETTING: Research laboratory of an academic institution. SUBJECTS: Primary human hepatocytes and human hepatoma HepaRG cells. INTERVENTIONS: Primary human hepatocytes and human hepatoma HepaRG cells were exposed to lipopolysaccharide to evaluate effects of epinephrine and several other compounds (norepinephrine, dobutamine, dopamine, dopexamine, phenylephrine, clonidine, salbutamol, and vasopressin). Markers of inflammation (interleukin-6, C-reactive protein) and drug metabolism (cytochrome P450 [CYP] 3A4, CYP2B6, CYP1A2, CYP2E1, constitutive androstane receptor, pregnane X receptor) were analyzed. MEASUREMENTS AND MAIN RESULTS: Transcripts of C-reactive protein and CYP3A4 were strongly increased and depressed respectively after a 24-hr treatment with 10 ng/mL lipopolysaccharide. Co-treatment with either of the catecholamines failed to reverse lipopolysaccharide effects, whereas when added alone, epinephrine, and to a lesser extent norepinephrine, salbutamol, and dobutamine, mimicked lipopolysaccharide effects. Suppression of CYP3A4 implicated beta-adrenergic receptors and was mediated through overproduction of interleukin-6. By contrast, vasopressin did not elicit an inflammatory response or modify CYP3A4 expression. CONCLUSIONS: Some catecholamines can induce an inflammatory response and exacerbate the hepatic dysfunction observed during sepsis, favoring the idea that catecholamines could alter the biotransformation of drugs metabolized by CYP3A4 and that alternative vasoactive agents, such as vasopressin, merit further investigation in septic shock patients. PMID- 18431273 TI - Differential gene profiling in acute lung injury identifies injury-specific gene expression. AB - OBJECTIVES: Acute lung injury can result from distinct insults, such as sepsis, ischemia-reperfusion, and ventilator-induced lung injury. Physiologic and morphologic manifestations of disparate forms of injury are often indistinguishable. We sought to demonstrate that acute lung injury resulting from distinct insults may lead to different gene expression profiles. DESIGN: Microarray analysis was used to examine early molecular events in lungs from three rat models of acute lung injury: lipopolysaccharide, hemorrhage shock/resuscitation, and high-volume ventilation. SETTING: University laboratory. SUBJECTS: Male Sprague-Dawley rats (body weight, 300-350 g). INTERVENTIONS: Rats were subjected to hemorrhagic shock or lipopolysaccharide followed by resuscitation or were subjected to sham operation. First hit was followed by ventilation with either low (6 mL/kg) or high (12 mL/kg) tidal volume for 4 hrs. MEASUREMENTS AND MAIN RESULTS: Physiologic and morphologic variables were assessed. Total RNA was hybridized to Affymetrix chips. Bioconductor was used to identify significantly altered genes. Functional enrichment predictions were performed in Gene Ontology Tree Machine. Confirmation studies included real-time polymerase chain reaction, Western blots, and immunohistochemistry. Physiologic and morphologic variables were noncontributory in determining the cause of acute lung injury. In contrast, molecular analysis revealed unique gene expression patterns that characterized exposure to lipopolysaccharide and high-volume ventilation. We used hypergeometric probability to demonstrate that specific functional enrichment groups were regulated by biochemical vs. biophysical factors. Genes stimulated by lipopolysaccharide were involved in metabolism, defense response, immune cell proliferation, differentiation and migration, and cell death. In contrast, high-volume ventilation led to the regulation of genes involved primarily in organogenesis, morphogenesis, cell cycle, proliferation, and differentiation. CONCLUSIONS: These results demonstrate the application of functional genomics to the molecular "fingerprinting" of acute lung injury and the potential for decoupling biophysical from biochemical injury. PMID- 18431274 TI - Redox status of acute pancreatitis as measured by cyclic voltammetry: initial rodent studies to assess disease severity. AB - OBJECTIVE: To determine whether serum antioxidant capacity as measured by the electrochemical technique cyclic voltammetry could be used to resolve differences in the severity of an inflammatory disease such as acute pancreatitis. DESIGN: Experimental animal study. SETTING: Animal laboratory, Department of Surgery, Faculty of Medical and Health Sciences, University of Auckland, New Zealand. SUBJECTS: Male Wistar rats. INTERVENTIONS: A total of 48 inbred male Wistar rats were studied in five experimental groups. Group 1 (baseline reference, immediate euthanasia, n = 14) had no surgical intervention. Group 2 (sham, n = 9) had identical surgical procedures to the pancreatitis groups except for the intraductal infusion. Groups 3-5 (n = 9, n = 10, and n = 6, respectively) had acute pancreatitis induced by the pancreatic intraductal infusion of 3%, 4%, or 5% sodium taurocholate, respectively. Groups 2-5 were killed after 12 hrs. MEASUREMENTS AND MAIN RESULTS: Cyclic voltammetry involves scanning the voltage of a working electrode while recording the anodic current produced as the low molecular weight antioxidants in the solution are oxidized on the surface of the working electrode. The current produced is proportional to the combined concentration of the antioxidants. There was a significant positive correlation of the first cyclic voltammetric peak maximum with pancreatic histologic severity (Spearman's r = .51, p = .007) and with a number of other markers of systemic severity, notably bicarbonate (r = -.57, p = .002), base excess (r = -.65, p < .001), urea (r = .68, p < .001), and calcium (r = -.60, p = .008). The first cyclic voltammetric peak maximum was superior at indicating the severity of the disease state compared with a standard method of total antioxidant capacity measurement. CONCLUSIONS: In experimental pancreatitis, the first cyclic voltammetric peak maximum showed significant correlations with histologic and systemic indices of severity. Further clinical studies are now needed to define the role of cyclic voltammetry in monitoring the progression of this and other severe illness in the critical care setting. PMID- 18431275 TI - Autologous transplantation of bone marrow-derived endothelial progenitor cells attenuates monocrotaline-induced pulmonary arterial hypertension in rats. AB - OBJECTIVES: Bone marrow-derived endothelial progenitor cells have been shown to circulate to damaged vascular endothelium and differentiate into mature endothelial cells. This study investigated whether bone marrow-derived endothelial progenitor cell therapy ameliorates monocrotaline (MCT)-induced pulmonary arterial hypertension in a rat model. DESIGN: Male Sprague-Dawley rats were randomized to receive MCT (75 mg/kg) only (group 1), MCT plus autologous bone marrow-derived endothelial progenitor cell (1.2 x 10(6) cells) transplantation (group 2), and saline injection only (group 3). Mononuclear cells were obtained from femoral bone marrow of group 2 rats and isolated by Ficoll gradient centrifugation. The cells were cultured for 21 days in endothelial culture medium. SETTING: An animal research laboratory at Kaohsiung Chang Gung Memorial Hospital. MEASUREMENTS: Hemodynamics, ventricular weight, expressions of connexin43, endothelial nitric oxide synthase messenger RNA gene, Bcl-2, and number of alveolar sacs and small lung arterioles were measured. RESULTS: Hemodynamic measurements on day 28 after MCT treatment revealed the development of significantly increased pulmonary arterial hypertension in MCT-treated groups (p < .0001). The bone marrow-derived endothelial progenitor cells were intravenously transplanted in group 2 on day 28 after MCT-induced pulmonary arterial hypertension. By day 90 after MCT treatment, the right ventricular systolic blood pressure and right ventricular hypertrophy were significantly increased in group 1 compared with groups 2 and 3 (all p values <.01). In addition, connexin43 and endothelial nitric oxide synthase messenger RNA gene expressions of lung and right ventricle and Bcl-2 protein expression of right ventricle were significantly lower in group 1 than in groups 2 and 3 (all p values <.01). Furthermore, the number of alveolar sacs and small lung arterioles were significantly lower in group 1 than in groups 2 and 3 (all p values <.01). CONCLUSIONS: Autologous bone marrow-derived endothelial progenitor cell transplantation effectively ameliorates MCT-induced pulmonary arterial hypertension. PMID- 18431276 TI - Prearrest administration of low-molecular-weight heparin in porcine cardiac arrest: hemodynamic effects and resuscitability. AB - OBJECTIVE: Both animal and human studies demonstrate activation of coagulation during cardiac arrest. Prearrest anticoagulation is used routinely in many experimental studies. We studied the hemodynamic effects of prearrest anticoagulation with a low-molecular-weight heparin suitable for clinical use during cardiopulmonary resuscitation in pigs. DESIGN: Randomized and blinded experimental animal study. SETTING: University hospital-affiliated research laboratory. SUBJECTS: Sixteen female domestic pigs. INTERVENTIONS: Three minutes before electrically induced ventricular fibrillation, enoxaparin 1 mg/kg or physiologic saline was blinded and administered intravenously. After 10 mins of untreated ventricular fibrillation, advanced cardiac life support was initiated with continuous mechanical chest compressions and interposed manual ventilation with 100% oxygen. Epinephrine was administered after 2 mins of advanced cardiac life support followed by attempted defibrillation 1 min thereafter. Advanced cardiac life support was continued for 10 mins following international guidelines. Electrocardiogram was recorded continuously and ventricular fibrillation waveform was analyzed (median slope). Animals with return of spontaneous circulation were observed for ten more minutes. Blood specimens were drawn for analysis of coagulation activation (thrombin-antithrombin complex) and drug effect (anti-factor Xa activity). MEASUREMENTS AND MAIN RESULTS: Six of eight (75%) pigs in each group achieved return of spontaneous circulation. Thrombin-antithrombin complex levels were significantly lower in pigs that received enoxaparin. There was no significant difference either in measured hemodynamics between the groups during advanced cardiac life support and after return of spontaneous circulation or in median slope values during ventricular fibrillation. Epinephrine caused a significant decrease in femoral and increase in cerebral cortical blood flow with no difference between the groups. CONCLUSIONS: Prearrest anticoagulation with enoxaparin did not influence either hemodynamics during advanced cardiac life support and after return of spontaneous circulation or the frequency of return of spontaneous circulation in porcine cardiac arrest. PMID- 18431277 TI - Combined effects of propofol and dexamethasone on rats with endotoxemia. AB - OBJECTIVE: To clarify the effect of combined treatment with propofol and dexamethasone on hemodynamics, organ injury, and survival rate in rats with endotoxemia. DESIGN: Randomized, prospective animal experiment. SETTING: Academic research laboratory. SUBJECTS: Male Wistar rats. INTERVENTIONS: Rats were divided into five groups: a control group, a group of conscious rats infused with Escherichia coli lipopolysaccharide, two groups of lipopolysaccharide rats treated with either propofol or dexamethasone, and a group of lipopolysaccharide rats with combined treatment of propofol and dexamethasone. MEASUREMENTS AND MAIN RESULTS: All hemodynamic and biochemical variables were measured during the 6-hr observation. Propofol plus dexamethasone attenuated hypotension and delayed hypoglycemia and metabolic acidosis caused by coadministration of E. coli lipopolysaccharide. In addition, propofol plus dexamethasone attenuated the lipopolysaccharide-induced multiple organ dysfunctions, such as lung, liver, and kidney. The increases in serum tumor necrosis factor-alpha, tissue nitric oxide, and superoxide anion levels were attenuated by propofol plus dexamethasone in lipopolysaccharide rats. Microscopic findings confirmed that propofol plus dexamethasone attenuated the substantial swelling and cell infiltration in lung and kidney caused by endotoxin. The 22-hr survival rate after endotoxin injection was markedly increased in lipopolysaccharide rats with combined treatment compared with the lipopolysaccharide rats (80% vs. 0%). CONCLUSIONS: The combined treatment with propofol plus dexamethasone reduced mortality rate and attenuated organ injury in conscious rats treated with lipopolysaccharide. These protective effects may be associated with their anti-inflammatory capacity and antioxidant activity. PMID- 18431278 TI - Therapeutic hypothermia with a novel surface cooling device improves neurologic outcome after prolonged cardiac arrest in swine. AB - OBJECTIVE: Devices for rapid induction of mild hypothermia after cardiac arrest are needed. We hypothesized that the Life Recovery Systems' ThermoSuit System provides effective core cooling by pumping ice water over the skin surface and improves neurologic outcome after prolonged cardiac arrest. DESIGN: Prospective experimental study. SETTING: University research laboratory. SUBJECTS: Large White breed pigs (29 to 35 kg). INTERVENTIONS: Swine were anesthetized and mechanically ventilated. Ten minutes of untreated ventricular fibrillation, 3 mins of basic life support, and 5 mins of advanced cardiac life support, including two 0.4 IU/kg doses of vasopressin, were followed by up to three countershocks. After restoration of spontaneous circulation, swine were randomized to two groups (normothermic control, hypothermia). The hypothermia group was cooled from a pulmonary artery temperature of 38.5 +/- 0.5 degrees C to 33.0 degrees C and kept for 14 hrs. At day 9 of the experiment, overall performance categories scores (1, normal; 2, slightly disabled; 3, severely disabled; 4, comatose; 5, dead, brain dead) and neurologic deficit scores (0%, normal; 100%, brain dead) were assessed. Data are presented as median and interquartile range; group comparison was done with a Mann-Whitney U test. MEASUREMENTS AND MAIN RESULTS: In total, 16 of 22 animals were randomized. Time to target temperature in the hypothermia group (n = 8) was 9.0 (5.3-11.9) mins (cooling rate 0.4 [0.3-0.8] degrees C/min), and all animals achieved an overall performance categories score of 1. In the control group, one swine achieved an overall performance categories score of 1, three achieved a score of 2, and four achieved a score of 3 (p = .002). Neurologic deficit score was 0% (0%-4%) in the hypothermia group and 39% (19%-55%) in the control group (p = .001). No harmful side effects could be observed. CONCLUSIONS: The Life Recovery Systems' ThermoSuit System rapidly and safely induced mild therapeutic hypothermia. Hypothermia improved neurologic outcome in swine after cardiac arrest as compared with normothermia. Further studies are warranted to compare the device with established cooling methods. PMID- 18431279 TI - Electrical impedance tomography compared with thoracic computed tomography during a slow inflation maneuver in experimental models of lung injury. AB - OBJECTIVE: To determine the validity of functional electric impedance tomography to monitor regional ventilation distribution in experimental acute lung injury, and to develop a simple electric impedance tomography index detecting alveolar recruitment. DESIGN: Randomized prospective experimental study. SETTING: Academic research laboratory. SUBJECTS: Sixteen anesthetized, tracheotomized, and mechanically ventilated pigs. INTERVENTIONS: Acute lung injury was induced either by acid aspiration (direct acute lung injury) or by abdominal hypertension plus oleic acid injection (indirect acute lung injury) in ten pigs. Six pigs with normal lungs were studied as a control group and with endotracheal suction related atelectasis. After 4 hrs of mechanical ventilation, a slow inflation was performed. MEASUREMENTS AND MAIN RESULTS: During slow inflation, simultaneous measurements of regional ventilation by electric impedance tomography and dynamic computed tomography were highly correlated in quadrants of a transversal thoracic plane (r2 = .63-.88, p < .0001, bias <5%) in both direct and indirect acute lung injury. Variability between methods was lower in direct than indirect acute lung injury (11 +/- 2% vs. 18 +/- 3%, respectively, p < .05). Electric impedance tomography indexes to detect alveolar recruitment were determined by mathematical curve analysis of regional impedance time curves. Empirical tests of different methods revealed that regional ventilation delay, that is, time delay of regional impedance time curve to reach a threshold, correlated well with recruited volume as measured by CT (r2 = .63). Correlation coefficients in subgroups were r2 = .71 and r2 = .48 in pigs with normal lungs with and without closed suction related atelectasis and r2 = .79 in pigs subject to indirect acute lung injury, respectively, whereas no significant correlation was found in pigs undergoing direct acute lung injury. CONCLUSIONS: Electric impedance tomography allows assessment of regional ventilation distribution and recruitment in experimental models of direct and indirect acute lung injury as well as normal lungs. Except for pigs with direct acute lung injury, regional ventilation delay determined during a slow inflation from impedance time curves appears to be a simple index for clinical monitoring of alveolar recruitment. PMID- 18431280 TI - Systemic inflammatory response syndrome increases immobility-induced neuromuscular weakness. AB - OBJECTIVE: Inflammation and immobility are comorbid etiological factors inducing muscle weakness in critically ill patients. This study establishes a rat model to examine the effect of inflammation and immobilization alone and in combination on muscle contraction, histology, and acetylcholine receptor regulation. DESIGN: Prospective, randomized, experimental study. SETTING: Animal laboratory of a university hospital. SUBJECTS: Sprague-Dawley rats. INTERVENTIONS: To produce systemic inflammation, rats (n = 34) received three consecutive intravenous injections of Corynebacterium parvum on days 0, 4, and 8. Control rats (n = 21) received saline. Both groups were further divided to have one hind limb either immobilized by pinning of knee and ankle joints or sham-immobilized (surgical leg). The contralateral nonsurgical leg of each animal served as control (nonsurgical leg). MEASUREMENTS AND MAIN RESULTS: After 12 days, body weight and muscle mass were significantly reduced in all C. parvum animals compared with saline-injected rats. Immobilization led to local muscle atrophy. Normalized to muscle mass, tetanic contraction was reduced in the surgical leg after immobilization (7.64 +/- 1.91 N/g) and after inflammation (8.71 +/- 2.0 N/g; both p < .05 vs. sham immobilization and saline injection, 11.03 +/- 2.26 N/g). Histology showed an increase in inflammatory cells in all C. parvum-injected animals. Immobilization in combination with C. parvum injection had an additive effect on inflammation. Acetylcholine receptors were increased in immobilized muscles and in all muscles of C. parvum-injected animals. CONCLUSIONS: The muscle weakness in critically ill patients can be replicated in our novel rat model. Inflammation and immobilization independently lead to muscle weakness. PMID- 18431281 TI - NKCC1-mediated traumatic brain injury-induced brain edema and neuron death via Raf/MEK/MAPK cascade. AB - OBJECTIVE: Brain edema is one of the characteristic features of patients with severe traumatic brain injury. The aim of this study was to examine the effects of Na+-K+-2Cl- co-transporter on traumatic brain injury-induced brain edema and neuron damage and to elucidate the relationship between Na+-K+-2Cl- co transporter and mitogen-activated protein kinase (MAPK) cascade. DESIGN: Laboratory investigation. SETTING: University research laboratory. SUBJECTS: Male Wistar rats weighing 350-400 g. INTERVENTIONS: Anesthetized animals were subjected to a weight-drop device (450-g weight, 1.8-m height) to induce traumatic brain injury. MEASUREMENTS AND MAIN RESULTS: The expression of Na+-K+ 2Cl- co-transporter and phosphorylation of MAPK cascade were determined by Western blot test. We also analyzed the degree of brain edema and neuronal damage in this study. We found that the messenger RNA and protein of Na+-K+-2Cl- co transporter were up-regulated mainly in hippocampus neurons from 2 to 24 hrs after traumatic brain injury. After traumatic brain injury, animals displayed severe brain edema and neuron damage. The phosphorylation of extracellular signal regulated kinase, MAPK kinase, and Raf also was significantly elevated after traumatic brain injury. Bumetanide (15.2 mg/kg), a specific Na+-K+-2Cl- co transporter inhibitor, significantly attenuated the neuronal damage and brain edema after traumatic brain injury by decreasing the phosphorylation of Raf/MEK/ERK cascade proteins. CONCLUSIONS: The present study suggests that Na+-K+ 2Cl- co-transporter plays an important role in TBI-induced brain edema and neuronal damage via activation of MAPK cascade. PMID- 18431282 TI - Relationship between global end-diastolic volume and cardiac output in critically ill infants and children. AB - OBJECTIVE: The objective of this study was to investigate possible correlations between the preload index global end-diastolic volume (GEDV) and the indexes of cardiac function, cardiac index, and stroke volume index in critically ill pediatric patients. The aim was to evaluate whether GEDV may help in the decision making process concerning volume loading. DESIGN: Prospective clinical study. SETTING: Pediatric intensive care unit of the Bambino Gesu Children's Research Hospital. PATIENTS: Seventy patients, 40 male and 30 female, mean age 62 +/- 41 months (range 5-156 months), divided into six groups: group A, hemorrhagic shock, ten cases; group B, head injury, 21 cases; group C, septic shock, ten cases; group D, encephalitis, ten cases; group E, respiratory failure, nine cases; group F, cardiogenic shock, ten cases. INTERVENTIONS: All patients received volumetric hemodynamic monitoring following initial resuscitation and every 4 hrs thereafter or whenever a hemodynamic deterioration was suspected. During the cumulative in hospital stay, a total 1,184 sets of measurements were done. MEASUREMENTS AND MAIN RESULTS: Findings are consistent with a statistically significant linear correlation of GEDV with cardiac index and stroke volume index in hemorrhagic shock (group A) (R2 = .647, p < .0001; R2 = .738, p < .0001) and cardiogenic shock (group F) (R2 = .645, p < .0001; R2 = .841, p < .0001). CONCLUSIONS: GEDV may potentially be a useful guide to treatment in preload-dependent conditions, such as hemorrhagic and cardiogenic shock. In the other groups where there is little relationship between preload and cardiac function indexes, the influence of non-preload-dependent mechanisms on cardiac output is certainly more significant. PMID- 18431283 TI - Educational interventions for prevention of healthcare-associated infection: a systematic review. AB - BACKGROUND: Healthcare-associated infections (HCAIs) are associated with considerable morbidity and mortality. Education of healthcare providers is a fundamental measure to prevent HCAI. OBJECTIVE: To perform a systematic review to determine the effect of educational strategies of healthcare providers for reducing HCAI. DATA SOURCE: Multiple computerized databases for the years 1966 to November 1, 2006, supplemented by manual searches for relevant articles. STUDY SELECTION: English-language controlled studies and randomized trials that included an educational intervention and provided data on the incidence of one or more kinds of HCAIs were included. DATA EXTRACTION: Data were extracted on study design, patient population, type of intensive care unit, details of the educational intervention, target group for intervention, incidence of HCAI, duration of follow-up, and costs of intervention. Both investigators abstracted data using a standard data abstraction form; study quality was also assessed. DATA SYNTHESIS: A total of 26 studies used a number of different educational programs targeting varied study populations of healthcare providers to determine their effect on HCAI rates. Most were pre-post intervention studies and were implemented in the intensive care setting. There was a statistically significant decrease in infection rates after intervention in 21 studies, with risk ratios ranging from 0 to 0.79. The beneficial effect of education was apparent in teaching and nonteaching institutions and in lesser-developed countries and developed nations. LIMITATIONS: Only English language studies were included. Because of the study designs and limitations of the individual studies, a causal association between educational interventions and reduced HCAI rates cannot be made. CONCLUSIONS: The implementation of educational interventions may reduce HCAI considerably. Cluster randomized trials using validated educational interventions and costing methods are recommended to determine the independent effect of education on reducing HCAI and the cost-savings that may be realized with this approach. PMID- 18431284 TI - Procalcitonin assay in systemic inflammation, infection, and sepsis: clinical utility and limitations. AB - OBJECTIVE: The use of procalcitonin (ProCT) as a marker of several clinical conditions, in particular, systemic inflammation, infection, and sepsis, will be clarified, and its current limitations will be delineated. In particular, the need for a more sensitive assay will be emphasized. For these purposes, the medical literature comprising clinical studies pertaining to the measurement of serum ProCT in various clinical settings was examined. DATA SOURCE AND SELECTION: A PubMed search (1965 through November 2007) was conducted, including manual cross-referencing. Pertinent complete publications were obtained using the MeSH terms procalcitonin, C-reactive protein, sepsis, and biological markers. Textbook chapters were also read and extracted. DATA EXTRACTION AND SYNTHESIS: Available clinical and other patient data from these sources were reviewed, including any data relating to precipitating factors, clinical findings, associated illnesses, and patient outcome. Published data concerning sensitivity, specificity, and reproducibility of ProCT assays were reviewed. CONCLUSIONS: Based on available data, the measurement of serum ProCT has definite utility as a marker of severe systemic inflammation, infection, and sepsis. However, publications concerning its diagnostic and prognostic utility are contradictory. In addition, patient characteristics and clinical settings vary markedly, and the data have been difficult to interpret and often extrapolated inappropriately to clinical usage. Furthermore, attempts at meta-analyses are greatly compromised by the divergent circumstances of reported studies and by the sparsity and different timing of the ProCT assays. Although a high ProCT commonly occurs in infection, it is also elevated in some noninfectious conditions. Thus, the test is not a specific indicator of either infection or sepsis. Moreover, in any individual patient, the precipitating cause of an illness, the clinical milieu, and complicating conditions may render tenuous any reliable estimations of severity or prognosis. It also is apparent that even a febrile septic patient with documented bacteremia may not necessarily have a serum ProCT that is elevated above the limit of functional sensitivity of the assay. In this regard, the most commonly applied assay (i.e., LUMItest) is insufficiently sensitive to detect potentially important mild elevations or trends. Clinical studies with a more sensitive ProCT assay that is capable of rapid and practicable day-to-day monitoring are needed and shortly may be available. In addition, investigations showing that ProCT and its related peptides may have mediator relevance point to the need for evaluating therapeutic countermeasures and studying the pathophysiologic effect of hyperprocalcitonemia in serious infection and sepsis. PMID- 18431285 TI - Recommendations for end-of-life care in the intensive care unit: a consensus statement by the American College [corrected] of Critical Care Medicine. AB - BACKGROUND: These recommendations have been developed to improve the care of intensive care unit (ICU) patients during the dying process. The recommendations build on those published in 2003 and highlight recent developments in the field from a U.S. perspective. They do not use an evidence grading system because most of the recommendations are based on ethical and legal principles that are not derived from empirically based evidence. PRINCIPAL FINDINGS: Family-centered care, which emphasizes the importance of the social structure within which patients are embedded, has emerged as a comprehensive ideal for managing end-of life care in the ICU. ICU clinicians should be competent in all aspects of this care, including the practical and ethical aspects of withdrawing different modalities of life-sustaining treatment and the use of sedatives, analgesics, and nonpharmacologic approaches to easing the suffering of the dying process. Several key ethical concepts play a foundational role in guiding end-of-life care, including the distinctions between withholding and withdrawing treatments, between actions of killing and allowing to die, and between consequences that are intended vs. those that are merely foreseen (the doctrine of double effect). Improved communication with the family has been shown to improve patient care and family outcomes. Other knowledge unique to end-of-life care includes principles for notifying families of a patient's death and compassionate approaches to discussing options for organ donation. End-of-life care continues even after the death of the patient, and ICUs should consider developing comprehensive bereavement programs to support both families and the needs of the clinical staff. Finally, a comprehensive agenda for improving end-of-life care in the ICU has been developed to guide research, quality improvement efforts, and educational curricula. CONCLUSIONS: End-of-life care is emerging as a comprehensive area of expertise in the ICU and demands the same high level of knowledge and competence as all other areas of ICU practice. PMID- 18431286 TI - Sepsis: time to reconsider the concept. AB - OBJECTIVE: To discuss the difficulty in using the concept of sepsis for clinical trials and propose new ways for designing future trials for severe infections. DESIGN: Short position statement. METHODS AND MAIN RESEARCH: Using a thorough evaluation of the recent literature in the field of severe sepsis and septic shock, the authors challenge the concept of sepsis as used in the past two decades and propose new ideas for designing future trials in this setting. The two main proposals are first to use a systematic assessment of the targeted inflammatory mediators when the study intends to counteract or replace those mediators (e.g., anti-tumor necrosis factor-alpha, activated protein C) and, second, to select more homogeneous populations, coming back to "precise infectious diseases," such as severe community-acquired pneumonia, severe peritonitis, or meningitis. CONCLUSIONS: The concept of sepsis has been useful to help clinicians to suspect and detect severe infections. Due to a considerable heterogeneity in the patients and type of infections included in the trials performed in the last two decades, it has not been useful in demonstrating the efficacy of new compounds. The authors propose a dramatic change in the design of future trials dealing with severe infections. PMID- 18431287 TI - Autonomic dysfunction predicts both 1- and 2-month mortality in middle-aged patients with multiple organ dysfunction syndrome. AB - OBJECTIVE: Multiple organ dysfunction syndrome (MODS) is a disease entity that carries a high mortality rate. It is characterized by a sequential failure of several organ systems after a trigger event, most commonly sepsis. There is increasing evidence that autonomic dysfunction may substantially contribute to the development of MODS. We recently characterized the spectrum of autonomic dysfunction by using heart rate variability in critically ill MODS patients and were able to show that autonomic dysfunction predicts 28-day mortality in MODS. The aim of the present study was evaluate whether autonomic dysfunction is also a predictor of 180-day and 365-day mortalities. DESIGN: Prospective cohort study. SETTING: Twelve-bed medical intensive care unit in a university center. PATIENTS: Ninety consecutively admitted score-defined MODS patients. INTERVENTIONS: We assessed heart rate variability as a marker of autonomic dysfunction. The patients were followed for 180- and 365-day mortalities. MEASUREMENTS AND MAIN RESULTS: We prospectively used the heart rate variability variable lnVLF, which predicted 28-day mortality best in the entire cohort of patients, for analysis of longer term mortality. The variable lnVLF was found to be useful for risk prediction for about 60 days, and then the survival curves became nearly parallel. CONCLUSIONS: Autonomic function of critically ill MODS patients is blunted, and this attenuation has prognostic implications not merely concerning 28-day mortality but also concerning longer term (about 2-month) mortality. PMID- 18431288 TI - The anatomical and functional shunt conundrum: what do we really know about the pathophysiology of acute respiratory distress syndrome? PMID- 18431289 TI - Intensive care unit readmission: the issue is safety not frequency. PMID- 18431290 TI - Soluble thrombomodulin: a sign of bad times. PMID- 18431291 TI - Reevaluating prognosis: cancer, septic shock, and changing outcomes. PMID- 18431292 TI - To sleep in an intensive care unit, perchance to heal. PMID- 18431293 TI - Importance of systems-based practice in achieving pharmacoeconomic benefits. PMID- 18431294 TI - Immunotherapy for overinflammatory states. PMID- 18431295 TI - Toward ethical best practices in community consultation for research conducted with waiver of informed consent. PMID- 18431296 TI - How can earlier antibiotic efficacy be provided for ventilator-associated pneumonia without promoting bacterial resistance? Is initial monotherapy or a combination of antibiotics the right answer? PMID- 18431297 TI - Natriuretic peptides, acute kidney injury, and clinical evidence. PMID- 18431298 TI - Filling the (strong ion) gap. PMID- 18431299 TI - Sepsis-induced left ventricular diastolic dysfunction: little time to relax. PMID- 18431300 TI - Finally, a tool for triage? PMID- 18431301 TI - Our failure to report failure. PMID- 18431302 TI - Noninvasive ventilation for patients near the end of life: what do we know and what do we need to know? PMID- 18431303 TI - Mannitol and hypertonic saline: going head to head. PMID- 18431304 TI - Memories of the intensive care unit. PMID- 18431305 TI - Placing bets with a full house. PMID- 18431306 TI - When mechanical ventilation mimics nature. PMID- 18431307 TI - Leptin and ghrelin: through thick and thin. PMID- 18431308 TI - Resuscitation end points in severe sepsis: central venous pressure, mean arterial pressure, mixed venous oxygen saturation, and... intra-abdominal pressure. PMID- 18431309 TI - Gene expression profiling in acute respiratory distress syndrome: pathways to future interventions. PMID- 18431310 TI - In vivo assessment of oxidative stress: a continuing challenge. PMID- 18431311 TI - Antithrombotic therapies for cardiac arrest: have we missed the mark? PMID- 18431312 TI - In quest of the optimal cooling device: isn't faster "too fast"? PMID- 18431313 TI - How to detect tidal recruitment and/or alveolar hyperinflation--computed tomography, electrical impedance tomography, or stress index? PMID- 18431314 TI - It's not only biochemistry: a possible pathogenetic role of immobilization/immobility in critical illness polyneuromyopathy. PMID- 18431315 TI - Just too good to be true (in the ICU)? Lower hospital mortality after pediatric hematopoietic stem cell transplantation. PMID- 18431316 TI - Measuring preload--one size doesn't necessarily fit all. PMID- 18431317 TI - Measures of parasympathetic function and risk stratification in critical care. PMID- 18431319 TI - Volume therapy and innate immune response during systemic inflammation or sepsis. PMID- 18431320 TI - [Hyponatremia: etiopathogenetic observations in the personal experiences]. AB - AIM: Hyponatremia, an electrolyte disturbance, can be due to loss of solutes or to an increase of ''free'' water concentration, predominantly caused by excessive antidiuretic hormone (ADH) secretion. When acute, hyponatremia is threatening, because it can cause cell swelling and severe neuromuscular symptoms (central pontine myelinolysis). METHODS: In a sample of 220 subjects with hyponatremia (123 males, 97 females) aged from 47 to 83, hospitalized in the Department of Clinic Medicines and Emergent Pathologies, from 2000 till today, age, serum and urinary sodium concentration, urine specific gravity, signs, symptoms and causes of electrolyte disturbance were evaluated. Student's t-test was applied for parametric spread data, Mann-Whitney Test for no parametric spread data, ANOVA test for statistic comparison between groups. RESULTS: Hyponatremia due to an increase of ''free'' water prevails in respect to hyponatremia due to loss of solutes (55.9% vs 44%). In the first case the prevalent causes are: decompensated hepatic cirrhosis (37.3%), heart failure (31.7%), glucosate solutions at 5% (17.8%) and drugs that can potentially increase ADH secretion (antidepressants, etc.) (13%), especially in elderly people. In the second case chronic diarrhea, vomiting (40%, 32.9%) and diuretic-induced hypernatriuresis (18.5%) prevail. Laboratory results follow the grade of efficient volemia, registering statistically significant differences between the two variants of hyponatremia. CONCLUSION: Different diagnosis and adequate treatment are essential when hyponatremia is acute. Particular attention must be reserved to elderly people with hyponatremia, in which the mechanisms of ''free'' water clearance are inefficient and such hypotonic glucosate solutions as medicines that increase ADH secretion (psychodrugs) must be administered with caution and under clinical control. PMID- 18431322 TI - The immunology of multiple sclerosis: disease mechanisms and therapeutic targets. AB - The disease mechanism of multiple sclerosis (MS) involves inflammation, demyelination and neurodegeneration. The relation between these components is not completely understood, but recent experiences with aggressive anti-inflammatory treatment suggest that inflammation drives neuronal damage in patients with relapsing remitting MS. Although infiltration of lymphocytes into the brain parenchyma was recognized as a key event in the pathogenesis of MS more than 120 years ago, important aspects of the mechanisms triggering and sustaining this immune response remain unknown. Furthermore, studies of MS lesions and evidence from therapeutic trials suggest that the disease mechanism may vary both throughout the disease course and between patients. The understanding of MS as an autoimmune disease targeting myelin proteins is shaped by the animal model experimental autoimmune encephalomyelitis (EAE), but translation from EAE to MS has proven to be difficult. Although both the EAE model and the prominent association to HLA class II molecules suggest a key role for CD4+ T helper cells, it is not known if or how their tolerance to myelin proteins or other putative autoantigens are broken in MS. This paper reviews some important concepts and controversies in the understanding of the immunological basis for MS and its treatment. PMID- 18431321 TI - Role of cytokine p40 family in multiple sclerosis. AB - Over the last couple of decades of neuro-immunological research, the p40 family of cytokines has emerged out as one of the most intriguing areas of interest because of multi-faceted roles of these cytokine in immune-modulation and inflammation. The IL-12, the most widely studied cytokine of this family, is well characterized for its Th1-favoring activity, and therefore plays a key role in the pathophysiology of Th1-mediated autoimmune diseases like multiple sclerosis (MS). On the other hand, the IL-23, another member of the p40 family with shared p40 subunit, drives polarization of Th17, a subset of T cell suspected to have a key role in the pathophysiology of MS and experimental allergic encephalomyelitis (EAE), poses a challenge to our current understanding of Th1/Th2 hypotheses in autoimmune diseases. However, the more puzzling issues, the researchers are currently confronted with, are the biological roles of other two members of this family, the p40 monomer and the p40(2), the homodimer. Predominance of the mRNA level of p40 over p35 in the central nervous system of EAE and MS suggests a possible involvement of p40 in the pathogenesis of MS. However, the distinctive biological role of monomeric and dimeric form of p40 is not clearly understood yet. Initially, it was thought that p402 does not have any biological activity and only involved in antagonizing bioactive IL-12 but according to recent evidences, both p402 and p40 appear to have a proinflammatory role, therefore might be a crucial molecule in the pathogenesis of MS. The current review focuses on biological function of p40 family of cytokines with particular emphasis on MS. PMID- 18431323 TI - Diagnosis and management of primary progressive multiple sclerosis. AB - Primary progressive multiple sclerosis (MS) is a chronic demyelinating degenerative disorder of the central nervous system. The most common presentation is with a spastic paraparesis, which may be asymmetrical. In contrast to relapsing remitting MS, discrete attacks are not a characteristic feature and the temporal course is of gradual symptomatic deterioration. The current diagnostic criteria are based on this clinical phenotype, with supportive evidence from magnetic resonance imaging, and examination of cerebrospinal fluid and visual evoked potentials in some cases. At present, there is no effective disease modifying therapy, but a wide range of symptomatic treatments are available. These may be of great benefit to individual patients and include pharmacological measures, multidisciplinary therapist input and neurorehabilitation. New treatments which target neurodegeneration and promote brain repair are required, and research in these areas offers hope for the future. PMID- 18431324 TI - Physical activity and its measurement and determinants in multiple sclerosis. AB - This literature review provides a description of research on physical activity behavior and its measurement and determinants in persons affected by multiple sclerosis (MS). Overall, there is fairly strong and consistent evidence that individuals with MS are largely inactive compared with non-diseased populations, and this rate of inactivity is alarming given the prevalence of inactivity among the general population. Importantly, there has been some concern with the measurement of physical activity in persons with MS, but this concern recently has been allayed by the systematic provision of evidence supporting the validity of both objective and self-report instruments in individuals with MS. Such evidence has enhanced the capacity of researchers for examining determinants of physical activity in persons with MS, and the emerging research has highlighted the role of symptoms in explaining inactivity in this population. As this body of research expands further, there will be a better basis for conducting targeted and well-designed interventions for promoting physical activity among those with MS. Efforts toward increasing physical activity in those with MS will go along way in promoting the many physiological and psychological consequences of an active lifestyle. PMID- 18431325 TI - Osteoporosis: clinical features. AB - Clinical features associated with osteoporotic fractures include increased morbidity (pain, physical impairment, decreased quality of life), increased risk for new fractures (even within short-term) and increased mortality. Readily recognizable clinical features that indicate a high risk for fracture include age, gender, low body weight, history of fracture, familial history of fracture, severe immobilization, smoking, rheumatoid arthritis, use of glucocorticoids and clinical risks for falls. In addition, many patients with fractures and osteoporosis have pre-existing contributors to secondary osteoporosis, many of which are correctable. PMID- 18431326 TI - Contrast media induced nephropathy: definition, incidence, outcome, pathophysiology, risk factors and prevention. AB - Iodanated contrast media related nephrotoxicity is a common phenomenon, which has been known for over five decades now and is widely termed contrast media induced nephropathy (CIN). CIN is one of the most common reasons for hospital-acquired acute kidney injury. The incidence of CIN varies widely, depending on the diagnostic criteria and on the individual risk factors for CIN. CIN is associated with a significant increase in mortality. The clinical features and the histological morphology of CIN are well characterized, whereas the precise mechanisms of renal injury during CIN are not clear. Most likely a combination of different pathomechanisms is involved: contrast media induced reduction of renal perfusion, reduction of tubular flow and direct tubular toxicity leading to a decrease of glomerular filtration rate. Several risk factors for the development of CIN exist, which can be divided into patient related and non-patient related risk factors as well as in modifiable and non-modifiable risk factors. Among them pre-existing renal failure is the most critical parameter. There is no effective treatment for CIN, however, because the development of CIN is predictable, preventional strategies for CIN have been developed. They can be divided into four different categories: volume expansion before, during and after contrast media administration, pharmacological strategies to prevent reduction of renal perfusion, reduction of tubular flow and direct tubular toxicity, renal replacement therapy, and selection of contrast media. Among them, volume expansion is the most critical measure for reducing CIN and should be used in all patients undergoing contrast media procedures. PMID- 18431327 TI - [Polimorphonuclear leukocytes and diabetes mellitus]. AB - Diabetic subjects have a higher infective risk than healthy people, with more frequent and severe infections. This predisposition to infections is determined by hyperglycemia, microangiopathy and altered immune system. In particular, there is a polymorphonuclear leukocytes disfunction including chemotaxis, phagocytosis, bacterial killing and cellular activation by infective stimulus. These alterations are due to abnormal properties of polymorphonuclear leukocytes (PMN) in diabetic patients. Several parameters like phagocytosis of bacterial cells, chemiluminescence during oxidative burst and cell membrane deformability are related to glycaemia and glycated hemoglobin. Recent acquisitions show an altered integrin pattern on diabetics PMN, at baseline and after in vitro stimulation with soluble stimulus like fMLP or PMA. This could influence the interactions between PMN and endothelial cells and the diapedesis. Receptorial alterations on PMN surface may be ascribed to the abnormalities of the cytoscheleton, of the endocytosis and of the transduction mechanism, due to hyperglycemia. PMID- 18431328 TI - Gout and related morbid conditions: pharmacological and SPA therapy. AB - Gouty arthritis is estimated to be the most frequent manifestation of inflammatory arthritis in men aged over 40. Hyperuricemia occurs because of both exogenous and genetic factors, which are particularly influential in some populations such as Taiwan aborigines. Current understanding of the disease etiopathogenesis, its clinical manifestations and the stages of its progression are presented here. The criteria for a correct diagnosis of the disease are also reported, pointing out how to distinguish gout from clinical events of different origin but with a very similar symptomatology. A distinction is made between the agents used to relieve the acute attack (colchicine, nonsteroidal anti inflammatory drugs, corticosteroids) and those used with the purpose of correcting hyperuricemia and preventing recurrences and complications (allopurinol, uricosurics). Mecha-nisms of action, administration routes, doses, side effects and contraindications of every drug are described. Besides pharmacological therapy, the importance and the efficacy of spa therapy is underlined. Finally, perspectives opened by gene therapy are mentioned. PMID- 18431329 TI - A singular case of granulomatous lesions of unknown significance. AB - The acronym used for granulomatous lesion of unknown significance, (GLUS), was first employed in 1990, to describe epithelioid-cell granulomas occasionally observed in biopsies from liver, lymph-nodes and other tissues without any apparent cause than thorough diagnostic examinations. Authors describe a case of a 51-year-old woman who was admitted to hospital because of fever, nodules in the liver and spleen, skin ulcerative lesions on the legs. The patient had a very long medical history; recurrent fever, liver and spleen enlargement have been lasting since 1975. Laboratory data showed erythrocyte sedimentation rate increase (125 mm 1 hour), anemia, (Hb=8.1 g/dL), an increase of alkaline phosphatase activity (328.4 mU/mL NV=80-275) and also an increase of policlonal gamma-globulins; patient's total proteins was 6.8 g/dL while their gamma globulins were 29.5%; IgG=2 260 mg/dL (NV=751-1 560 mg/dL), IgM=277 mg/dL (NV=46 230 mg/dL), IgA=405 mg/dL (NV=70-400 mg/dL). One culture of the blood and a needle aspiration specimen of liver nodule was positive for Staphylococcus coagulase negative. This case cannot be regarded as GLUS-syndrome because of the long time duration of the disease and other clinical features. The dramatic, even if transient, liver nodules improvement obtained by Linezolid therapy, and the well-being obtained by prolonged monociclin use, make the authors hypothesize that Staphylococcus coagulase negative could be the etiological agent of this granulomatous clinical picture. PMID- 18431330 TI - Acute diarrhea during azathioprine treatment of type I autoimmune hepatitis. AB - Sporadic descriptions of acute onset of watery diarrhea within a few hours to a few weeks azathioprine administration beginning have been reported, particularly in inflammatory bowel disease patients. This article reports the case of a woman treated with azathioprine because of type I autoimmune hepatitis, who developed acute watery diarrhea after more than two months of therapy. In two occasions the patient reassumed the drug and in a few hours diarrhea recurred. Subsequent 6 mercaptopurine treatment was well tolerated, suggesting that the previous side effect could be due to the nitroimidazole moiety of azathioprine. PMID- 18431331 TI - [The new hospital model established at the end of 19th Century and the professional nursing schools]. AB - The historical research in hospitals organisation began in Europe only after the II World War. In particular, studies regarding the nursing in Italy are very scarce and heavily influenced by the fragmented political and organisational management due to the historical divisions within Italy. All the same, the social and working conditions, at least in the north of Italy, were similar in this respect. The workforce up to the beginning of the 20th Century was equally divided between males and females. The female recruitment took place exclusively from the foundlings left in front of the hospital and raised within the hospital walls. The work was very hard with long working hours, under a total subordination to the religious administrators with miserable wage. These conditions remained unchanged until the second half of the 19th Century when, in central Europe, a change in the reorganisation of hospital management occurred, together with a cultural improvement in the nursing profession. This process of reforms had started in England some decades earlier from the reorganisation of nursing made by Florence Nightingale. After Italian post-unification the healthcare field was involved in the tentative to homologate the vast differences in the hospital organisation within the Italian states which also generated tension, bitterness and many political debates. The particular conditions created in the main hospital of Turin in the second half of the 19th Century, together with the political and social climate at the time and the reopening of debates between Socialist, Liberals and Catholics favoured the reorganizational improvement of the San Giovanni Battista hospital. Under this prospective, the foundation of the nursing school came into being, preceded by the intense control of a behavioral and ethical cleaning of the nurses, in particular the males. The availability of the hospital administrations in carrying out the recommendations of the medical staff in the modernisation and sanitary improvements and also in the quality of service was manifested by the decision to keep only the personnel who had obtained qualifications at the ''Nursing School''. This was also due to the financial support of the school itself. A further indication of appreciation for the school activity was the premature decision to extend the course to two years. This formative activity rapidly showed positive results, improving service quality and raising the professional ethics of the nurses. A few years after the foundation of the Nursing School in Turin, the school was contacted by numerous Italian hospital administrations to obtain information on the practices and organisational activities of the school. Thus, the process of professional training of nursing in Italy gradually started to take hold. At the same time, despite the diffusion of the hospital nursing schools, the experience of the Nightingale style of schooling in Italy never firmly established with success. The continuity of the professional instruction is instead to be noted in the transformation of the hospital boarding schools, exclusively for females, which in turn evolved into the classic Italian professional nursing school. PMID- 18431332 TI - The management of lower limb ischemia. PMID- 18431333 TI - Heparin-bonded expanded polytetrafluoroethylene grafts for infragenicular bypass in patients with critical limb ischemia: 2-year results. AB - AIM: The use of prosthetic grafts in below-knee (BK) bypasses may be necessary in patients with no available autologous vein and critical limb ischemia not amenable to angioplasty. Such conduits, however, have generally yielded disappointing results. METHODS: A new heparin-bonded expanded polytetrafluoroethylene graft (Gore-Tex Propaten Vascular Graft) designed to provide resistance to thrombosis may be associated with decreased early graft failure and increased patency. This graft was implanted in 27 limbs (26 patients; 18 men; mean age 71 years; Rutherford class 4 to 6 disease) in a BK femoropopliteal and femorodistal location, without perioperative complications and with immediate graft patency. RESULTS: During a mean follow-up time of 24 months, 4 cases of thrombosis occurred, all at least 6 months postoperatively: 2 cases resolved after fibrinolytic treatment, 1 required surgical revision and in 1 case, amputation was required because of a delay in seeking treatment for thrombosis. Two patients died of cardiac disease during follow-up. The 2-year primary and secondary patency rates for the BK bypasses were 85% and 93%, respectively; the limb-salvage rate was 96%. CONCLUSION: These results are encouraging for a prosthetic graft, especially in the light of the severity of the vascular disease in the limbs treated. PMID- 18431334 TI - Duplex-guided infrainguinal balloon angioplasty and stenting. A 4-year experience. AB - AIM: The traditional technique of infrainguinal arterial balloon angioplasties involves the use of fluoroscopy and contrast material. We performed these procedures under duplex guidance to eliminate radiation exposure and avoid nephrotoxic effect of contrast. METHODS: Over the last four years, 274 patients (59% males) with a mean age of 74+/-9 years (range 42-97 years) had a total of 360 attempted balloon angioplasties of the superficial femoral (SFA) and/or popliteal arteries under duplex guidance. Cannulation of common femoral artery, manipulation of the guidewire across the stenoses and/or occlusions of the SFA and/or popliteal artery, and balloon dilation were achieved with duplex guidance alone. Infrapopliteal angioplasties of 80 arteries were attempted in 54 cases (15% of all cases). RESULTS: Overall technical success for femoral-popliteal segment was 95% (342/360 cases) and 96% (77/80 cases) for infrapopliteal segment. CONCLUSION: Duplex guided balloon angioplasty and stent placement appears to be a safe and effective technique for treatment of femoral-popliteal and infrapopliteal arterial occlusive disease. PMID- 18431335 TI - Present and future of endovascular SFA treatment: stents, stent-grafts, drug coated balloons and drug coated stents. AB - The current evolution towards treating more complex femoropopliteal lesions as seen in the renewed TASC II recommendations clearly reflects the continuous evolutions in femoropopliteal stent design. Recent stent design improvements focus on decreasing stent fractures rates which can negatively impact patency rates. By rearranging strut alignment towards helical connecting bridges or full helical stent designs, stent designers have successfully improved the fracture resistance of their stents. The superior fracture resistance of the latest stent generation in combination with the production of long nitinol stents of up to 20 cm in length allow that more difficult and complex lesions can be treated endovascularly. The continuous perfection of nitinol stent platforms followed by the transfer of coronary technologies (e.g. active coatings) towards peripheral devices might broaden the indications for the minimal invasive strategy towards TransAtlantic Inter-Society Consensus (TASC) II C&D lesions. PMID- 18431336 TI - Recent advances in atherectomy and devices for treatment of infra-inguinal arterial occlusive disease. AB - The prevalence of peripheral arterial disease (PAD) continues to rise in an ever ageing society and consumes a significant part of health resources. Percutaneous revascularization has revolutionized the treatment of lower extremity peripheral vascular disease over the past 10 years. Additionally, novel devices have allowed improved endovascular treatment of femoropopliteal as well as infrapopliteal disease. Although percutaneous transluminal angioplasty (PTA) can be an effective modality for focal lesions in the iliac arteries, the results for complex infra inguinal arterial disease have been disappointing. One class of new technology has concentrated on debulking the plaque, while others focus to improve safety (distal embolic protection devices) or are directed to specific clinical challenges such as chronic total occlusions. However, the lack of uniform performance criteria and reporting standards for these and other devices has resulted in heterogeneous study end points, making comparative efficacy difficult. Here we review the current data for atherectomy and atheroablative technologies as well as other adjunctive devices in the treatment of lower extremity peripheral arterial disease. PMID- 18431337 TI - What is the evidence for the efficacy of cryoplasty? AB - Cryoplasty is a technique for treating vascular stenosis which combines balloon angioplasty with cold injury. The combination is proposed to reduce the incidence of restenosis by inhibition of neointimal hyperplasia. There have been several clinical studies which purport to show improved patency compared to conventional angioplasty. Unfortunately, these are not comparative or controlled studies and have not been performed, analyzed or reported in accordance with recognized reporting standards for peripheral vascular intervention. The studies on femoropopliteal disease have selected favourable patients. Of greatest concern is the use of surrogate endpoints in lieu of objective demonstration of vessel patency. Critical interpretation of the results fails to demonstrate any convincing superiority of cryoplasty compared to conventional balloon angioplasty. Where little difference in outcome exists between two techniques, a trial comparing them will require hundreds of patients to be sufficiently powered to demonstrate a benefit of one technique over the other. As cryoplasty is significantly more expensive than conventional angioplasty, the cost benefit ratio is unfavourable and such a trial is unlikely to occur. This article is a critical review of the technique of cryoplasty. The reader will be able to: describe expected outcomes from balloon angioplasty; describe the theoretical role for cold injury as a component of angioplasty; recognize the need to adhere to well defined standards when reporting the results of new techniques for treating vascular disease; critically review the results of cryoplasty; understand the limitations and relevance of the published clinical results of cryoplasty. PMID- 18431338 TI - Revascularization strategies in below the knee interventions. AB - Until recently the role of endovascular revascularization in the infrapopliteal arteries was limited. Endo-vascular management including percutaneous transluminal angioplasty and subintimal angioplasty is now being considered as a primary alternative in critical limb ischaemia revascularization with a decreasing number of major amputations (above the ankle) performed. Endovascular treatment has been shown to have reduced morbidity and hospital stay compared to revascularization surgery with greater cost-effectiveness and better patient quality of life compared with major amputation. PMID- 18431339 TI - Remote superficial femoral artery endarterectomy. AB - Remote superficial femoral artery endarterectomy (RSFAE) is an effective minimal invasive treatment modality of TransAtlantic Inter-Society Consensus (TASC) C and D atherosclerotic lesions of the superficial femoral artery (SFA) with at least equal patency rates as supragenicular synthetic bypass grafts. This procedure is performed through a single femoral arteriotomy and the intima core in the SFA is dissected using the Vollmar ring and the Mollring cutter devices, respectively. The intimal core distally of the transaction zone is secured by an expandable polytetrafluoroethylene-covered nitinol stent. By its minimal invasive character, RSFAE will lead to lower rate of postoperative complications and shorter hospital stay compared to supragenicular bypass graft surgery. Additional advantage in comparison with percutaneous procedures is the opportunity of open endarterectomy of the common femoral and/or profunda artery. Synthetic material will be avoided and vein will be preserved for possible future cardiovascular surgery. Reobstruction of the SFA tends to have, in contrast to bypass grafts, less severe symptoms due to preservation of collaterals and thereby lower amputation rate. Achilles heel of RSFAE is the relatively high percentage of first year restenosis due to neointimal hyperplasia. Strict follow-up at 3, 6 and 12 months is advised including duplex ultrasound. In case of symptomatic or asymptomatic hemodynamic restenosis (>50%) percutaneous transluminal angioplasty must be performed to improve long-term patency. The majority of reobstructions can be treated by endovascular means. New endovascular techniques, like balloon cryoplasty or drug eluting stents have to be studied in combination with RSFAE to optimize its technique and improve patency rates. PMID- 18431340 TI - Abdominal aortic aneurysm sac behavior following Cook Zenith graft implantation: a five-year follow-up assessment of 212 cases. AB - AIM: Aneurysm shrinkage is an expected outcome after stent-grafting for abdominal aortic aneurysm (AAA). A worrying problem following repair is progressive enlargement indicating persistent sac pressurization: in this setting not all grafts are equal. The Cook Zenith device (CZ) became available on the European market in 1999. While multicenter studies on the device have shown favorable clinical results at mid-term follow-up, few have focused on sac behavior. This study evaluated AAA sac behavior and predictive factors of its evolution by assessing the five-year results obtained with the CZ graft in a single institution experience. METHODS: All consecutive elective surgery patients treated with a CZ graft for infrarenal aortic or aortoiliac aneurysm repair from January 2000 to November 2004 in our institution were included prospectively in the study and followed at 1, 6, 12, 18, 24 months and yearly thereafter. Pre-, intra- and postoperative data were recorded in a computerized database. Computed tomography (CT) scans were reviewed by a senior radiologist to identify any abnormalities including endoleak and graft malfunction. Pre- and postoperative maximum sac diameters were derived from measurements of CT findings and then compared. A change of at least 8 mm in sac size was considered significant. Overall results are expressed according to the Committee on Reporting Standards of AAA treatment. Factors that may have influenced sac behavior were analyzed by dividing the patients into 3 groups according to whether the sac diameter remained unchanged (group 1), had increased (group 2) or regressed (group 3). Statistical analysis of the demographic and CT-scan data was then performed. RESULTS: The study sample was 212 consecutive patients (mean age 72.8+/-9.0 years); the mean aneurysm diameter before treatment was 55.5+/-9.8 mm. All stent grafts were successfully implanted. The 30-day mortality rate was 0.94% (2/212); the morbidity rate was 11.7% (25/212). The primary technical success rate was 93.40%; the assisted primary technical success and secondary technical success rates were 96.63% and 99.52%, respectively. The mean follow-up period was 17.7+/ 14.7 (1-60) months. The cumulative survival probability was 94%, 84.2%, and 72.9% at 12, 24, and 36 months, respectively. The endoleak-free survival probabilities at 12 and 24 months were 75.7% and 62.8%, respectively. The free of intervention survival rates were 82.1%, 68.9% and 60.6% at 12, 24 and 38 months, respectively. At five years follow-up, the overall clinical success rate was 49.5%. If changes in sac diameter occurred, they were noted at 13 months on average. Sac size remained unchanged in 115 (54.3%) patients (group 1), increased in 9 (4.2%) (group 2), and regressed in 88 (41.5%) (group 3). Neither preoperative patient demographics nor aneurysm characteristics were found to be predictive of sac behavior. Aortouniliac graft configuration was predictive of sac shrinkage (P=0.020). Endoleak was more frequent among patients in groups 1 (27/115; 23%) and 2 (5/9; 56%) than among those in group 3 (9/88;10%) (P=0.001). Reduction in aneurysm sac diameter was less marked in patients with any type of endoleak (P=0.0003). CONCLUSION: The CZ grafts offered satisfactory overall results up to five years of follow-up; nevertheless, sac diameter increased in 4.25% of patients. Endoleak was a predictive factor of a lack of sac shrinkage, while aortouniliac graft configuration was predictive of sac shrinkage. PMID- 18431341 TI - Carotid bifurcation atherosclerosis in the over-65s: a prevalence study. AB - AIM: The aim of this study was to determine the prevalence of carotid stenosis (CS) in the over-65 population segment residing in a catchment area (Gijon) served by the Asturias Health Service (Spain) as a necessary step in planning medical care for treating cerebrovascular disease in the elderly. METHODS: In this descriptive transversal study, 232 subjects (114 men and 118 women) randomly chosen from health card data underwent colour-flow duplex scanning of the supra aortic trunks. RESULTS: The prevalence of CS in this sample was 21.5%. When stratified by sex and age (65-74 and >75 years of age), the CS rate was 5 points higher in the older than in the younger group, and 4 points higher among males (23.6%) than among females (19.2%). CONCLUSION: Approximately one in every 5 subjects over 65 years of age presents with CS; CS prevalence was higher in the over-75s and among males, although the differences were not statistically significant. PMID- 18431342 TI - Systematic review and meta-analysis of endovenous radiofrequency obliteration, endovenous laser therapy, and foam sclerotherapy for primary varicosis. AB - AIM: Radiofrequency obliteration (RFO), endovenous laser therapy (EVLT) and foam sclerotherapy (FS) are potential treatments for varicose veins. A systematic review was undertaken to assess their safety and effectiveness and to compare these endoluminal therapeutic options with conventional ligation and vein stripping. METHODS: An electronic health database search was performed on all studies published between 1970 and 2007 describing RFO, EVLT, and FS for treating varicose veins. RESULTS: Twenty-nine EVLT studies, 32 RFO studies and 22 FS trials were included. RFO was associated with the worst short and long-term safety and efficacy results compared to EVLT and FS regarding ''complete occlusion at the end of follow-up'', ''phlebitis'', ''deep vein thrombosis'', and ''paraesthesia''. EVLT had the best results concerning the long-term effectiveness parameters for ''occlusion at the end of follow-up'' and ''recanalization, recurrence or development of new veins'', compared to RFO and FS. Foam sclerotherapy of varicose veins is associated with a higher recurrence rate in patients with saphenofemoral incompetence compared to the rates after EVLT or RFO treatment. CONCLUSION: EVLT, RFO, and FS seem to be safe and effective modalities with good short and mid-term RESULTS: Acquisition of comparative long-term and very long-term data on clinical efficacy (particularly with regard to the formation of recurrent varicose veins), safety, quality of life outcomes and costs is needed by large high-quality prospective randomized trials of endovenous techniques versus each other and versus surgery before considering endovenous techniques as the standard treatment. PMID- 18431343 TI - Advential cystic disease of the popliteal artery: experience of a single vascular and endovascular center. AB - Adventitial cystic disease (ACD) is an uncommon condition with only around 300 cases reported in the literature. ACD consists of a collection of gelatinous material within a cyst, that is adjacent or surrounding a vessel. In the last five years three cases of ACD have been observed: the first case was a 48-year old man, ex nicotine abuser, with a four-month history of progressive claudication; the second case was a 55-year-old man, ex nicotine abuser, with ischemic heart failure and a one-year history of progressive claudication; the third case was a 70-year-old man, with diabetes, dislipidemia and current nicotine abuser with a few-day history of acute pain in the right leg. In two cases the cysts were incised and the contents evacuated. The adventitia was repaired and the wound closed. The first patient is currently asymptomatic after four years from surgery. The second one, at 21 months, follow-up, presented newly severe claudication. Duplex ultrasound scan and computed tomography angiography demonstrated a cranial progression of the lesions. This lesion was treated with bare stent, with complete regression of the symptoms. The third patient was treated with interposition of an autogenous saphenous vein. The patient is asymptomatic at the three-month follow-up. Even if ACD is quite rare, it should be taken in consideration in young patients with severe claudication and no- or poor comorbidities. The best treatment is the incision of the cysts and the advential reconstruction. Short lesions can be treated with endovascular therapy. PMID- 18431344 TI - A rare vascular tumour with distinctive clinical findings. AB - Subungual glomus tumours are rare benign hamartomas arising from the arterial end of the glomus body (an arteriovenous shunt related to thermoregulation). A combination of clinical examination, Color-Doppler ultrasonography and magnetic resonance imaging (MRI) is used to support the diagnosis confirmed by histology. A 45 year old male patient with a three-year-old swelling of the proximal thumbnail presented himself at the Surgery Department. A slightly bluish nail plate and tenderness under the thumbnail were evident at clinical examination. He suffered the classic trilogy of sensitivity to pain, pressure and temperature of the thumb lesion. Ultrasound and MRI revealed the presence of a small oval-shaped mass with hypervascularisation. All these elements reinforced the assumed diagnosis of a glomic tumour. The patient underwent excisional surgery under local anaesthesia. Histological examination confirmed a typical glomus tumour. Postoperative follow-up was uneventful and all symptoms disappeared immediately after surgery. Whenever a patient presents suffering from a reddish-purple, tender lesion, usually located at the extremities and especially in the nail bed, with paroxysmal pain and cold-sensitivity, a glomus tumour should be investigated. Surgical excision is the treatment of choice and is commonly resolutive. PMID- 18431345 TI - Infected ruptured popliteal artery aneurysm by Listeria monocytogenes. A case report and review of the literatures. AB - We describe a case of a voluminous infected aneurysm of the popliteal artery, with Listeria monocytogenes (LM) associated with rupture, in a 72-year old man. After radical resection of the aneurysm a reconstruction was not necessary, because of the sufficient blood supply, due to the pre-existent good development of collateral circulation. The patient was discharged on the 12th postoperative day with primary healing of the wound and viable leg. Adequate antibiotic treatment was continued for 4 weeks. In the following 18 months the serial clinical examinations, laboratory tests and ultrasound scans have shown no evidence of reinfection. PMID- 18431346 TI - Protective effect of pyridoxal-5-phosphate (MC-1) on perioperative myocardial infarction is independent of aortic cross clamp time: results from the MEND-CABG trial. AB - AIM: Aortic cross-clamp time remains a significant marker of mortality and morbidity after coronary artery bypass graft (CABG) surgery. Pyridoxal-5 phosphate (MC-1), blocking purinergic receptors and intracellular influx of calcium, was shown to decrease the incidence of perioperative myocardial infarction in the prospective, randomized, double-blinded MC-1 to Eliminate Necrosis and Damage in CABG (MEND-CABG) clinical trial. METHODS: We studied the relationship between treatment with MC-1 and aortic cross-clamping relative to the incidence of cardiovascular (CV) death and myocardial infarction (MI) in the trial that enrolled 901 high-risk patients undergoing CABG with cardiopulmonary bypass. Patients were randomized to receive either placebo, MC-1 250 mg/day or MC 1 750 mg/day starting 3-10 h before CABG and continued for 30 days after surgery. Serial creatine kinase-myocardial band (CK-MB) determinations, ECGs and clinical evaluations were performed. RESULTS: Cross-clamping time increased the event rate of death and MI with an odds ratio (95% confidence interval) of 1.67 (1.17-2.37, P=0.0044). Treatment with MC-1 decreased the rate of events (P=0.0073) with odds ratios of 0.52 (0.31-0.88 for MC-1 250 mg/day versus placebo) and 0.48 (0.29-0.82 for MC-1 750 mg/day versus placebo). There was no interaction between cross-clamp time and treatment (P=0.61) on the occurrence of the combined endpoint. CONCLUSION: MC-1 decreased the incidence of CV death and MI (CK-MB >or=100 ng/mL) during the first 90 days after CABG in the MEND-CABG trial. Although longer aortic clamping time increased the risk of cardiovascular events, the protective effect of MC-1 was independent of ischemic time during CABG. PMID- 18431347 TI - Influence of angiotensin-I-converting-enzyme insertion/deletion gene polymorphism on perioperative hemodynamics after coronary bypass graft surgery. AB - AIM: The angiotensin I-converting enzyme insertion/ deletion polymorphism (ACE I/D), including three genotypes (II, ID, DD), with a known impact on midterm mortality and morbidity in patients after coronary artery bypass graft surgery (CABG), was studied. Since this polymorphism has been linked with increased vascular response to phenylephrine during cardiopulmonary bypass (CPB), we investigated its possible effect on perioperative hemodynamics in patients undergoing CABG. METHODS: Genotyping for the ACE-I/D was performed by polymerase chain reaction (PRC) amplification in 110 patients who underwent elective CABG with CPB. Patients were assigned to two groups according to their genotype (group II [II genotype] and group ID/DD [ID and DD genotypes]). Systemic hemodynamics were measured directly before and at 4 h, 9 h, and 19 h after CPB. RESULTS: Genotype distribution of ACE-I/D was 18%, 57%, and 25% in genotypes II, ID, and DD, respectively. The two groups were similar in age (group II: 66+/-6 years, group ID/DD: 66+/-8 years), body-mass-index (BMI) (group II: 28+/-2, group ID/DD: 29+/-5 kg/m2), male: female ratio (group II: 16: 4, group ID/DD: 63: 27) and Euroscore (group II: 3.1+/-1.9, group ID/DD: 3.5+/-2.1). There were no differences in mortality rate or perioperative systemic hemodynamics. The pulmonary vascular resistance before cardiopulmonary bypass was higher in the ID/DD genotypes than in the II genotypes (227+/-121 vs 297+/-169 dyn.s(-1).m2.cm( 5)). Four hours after CPB no difference remained; at 9 h after cardiopulmonary bypass there was a slight difference in pulmonary vascular resistance between the two groups (247+/-134 vs 290+/-117 dyn.s(-1).m2.cm(-5)) and a significant difference in pulmonary arterial pressure (19+/-6 vs 23+/-8); at 19 h after CPB the differences were no longer detectable. CONCLUSION: ACE-I/D had no influence on perioperative systemic hemodynamics. However, transitory differences in pulmonary hemodynamic were observed after CPB. These differences may have been due to changes in serum ACE activity during CPB. PMID- 18431348 TI - Is use of temporary pacing wires following coronary bypass surgery really necessary? AB - AIM: Temporary epicardial pacing wires (TEPW) which are routinely used after coronary bypass grafting may result in significant complications. We sought to identify variables that predict TEPW implantation and thereby limit their use. METHODS: This prospective study enrolled 564 patients (296 underwent coronary artery bypass grafting with cardiopulmonary bypass [ONCAB] and 268 underwent off pump coronary artery bypass grafting, OPCAB). TEPW were placed in patients with the intraoperative presence of one or more of the following criteria: sinus bradycardia, sinus arrest, nodal/junctional rhythms, atrioventricular block, bundle branch block, ventricular tachycardia, or onset of atrial fibrillation. RESULTS: Only 31 (5.5%) patients [ONCAB: 20 (6.8%) (ventricular: 14, bichamber: 6); OPCAB: 11 (4.1%) (ventricular: 9, bichamber: 2)] had temporary epicardial pacing wires implanted intraoperatively. Indications for using temporary epicardial pacing wires for ONCAB were sinus bradycardia (8), nodal/junctional rhythms (3), atrioventricular block (3), atrial fibrillation (4), and bundle branch block (2), and for OPCAB were sinus bradycardia (8), nodal/junctional rhythms (2), and atrioventricular block (1). Mean duration for pacing was 22.4 h for the ONCAB group and 11.3 h for the OPCAB group. There were no temporary epicardial pacing wires associated complications. One paced OPCAB patient required a permanent pacemaker and 2 non-paced OPCAB patients required transvenous pacing wires. Univariate and multivariate analyses were also conducted to determine risk factors for TEPW. CONCLUSION: TEPW implantation is overused in cardiac surgery and by identifying independent predictors for pacing we conclude that TEPW use should be limited to a select few. PMID- 18431349 TI - Connective tissue changes in a mouse model of vein graft disease. AB - AIM: The extracellular matrix plays an important physiological role in the architecture of the vascular wall. In arterialized vein grafts severe early changes, such as thrombosis and neointimal hyperplasia occur. Paclitaxel is in clinical use as antiproliferative coating of coronary stents. We aimed to investigate the early connective tissue changes in arterialized vein grafts and the influence of perivascular paclitaxel treatment in an in vivo model. METHODS: C57 black mice underwent interposition of the vena cava into the carotid artery. Neointimal hyperplasia, thrombosis, acid mucopolysaccharides (Alcian), collagen fibers (trichrome Masson), elastic fibers, and apoptosis rate (TUNEL) were quantified in paclitaxel treated veins and controls. RESULTS: In both, controls and paclitaxel treated vein grafts acid mucopolysaccharides and elastic fibers were found predominantly in the neointima, whereas collagen fibers were found mainly in the media and adventitia. At 4 weeks postoperatively the neointimal thickness in controls was 52 (13-130) microm, whereas in 0.6 mg/mL l paclitaxel treated veins it was 103 (43-318) microm (P=0.094). At 8 weeks postoperatively paclitaxel treated veins showed a significantly increased neointimal thickness of 136 (87-199) microm compared with 79 (62-146) microm in controls (P=0.032). There was no difference in apoptosis rate between the two groups (P=NS). Even with the lowest concentration of 0.008 mg/mL paclitaxel veins showed a neointimal thickness of 67 (46-205) microm at 4 weeks postoperatively (P=NS vs controls). CONCLUSION: Early vein graft disease is characterised by an accumulation of acid mucopolysaccharides and elastic fibers in the thickened neointima. Paclitaxel treatment increases the neointimal hyperplasia in mouse vein grafts in vivo. PMID- 18431350 TI - Heparin-coated extracorporeal circulation in combination with low dose systemic heparinization reduces early postoperative blood loss in cardiac surgery. AB - AIM: According to a recently performed meta-analysis, heparin-bonded circuits do not reduce blood loss in cardiac surgery patients compared to nonheparin-bonded circuits within the first 24 h postoperatively. We investigated the effects of heparin-coated circuits in combination with a reduced systemic heparin dose on early postoperative blood loss (first 12 h), platelet function, and postoperative complications. METHODS: Patients who underwent their first coronary artery bypass graft surgery were included in a randomized prospective study. Group A (n=149) was perfused with an uncoated extracorporeal circulation (ECC)-set and groups B (n=152) and C (n=149) with heparin-coated ECC-sets. In groups A and B, conventional dose systemic heparin was given, whereas group C received low dose systemic heparin. Blood loss was assessed within the first 12 h postoperatively. Moreover, biochemical parameters of pro-coagulant activity and immunological function were measured. RESULTS: None of the pro-coagulant activity markers and immunological parameters measured differed preoperatively or postoperatively between study groups. However, intraoperative platelet counts and maximal intraoperative concentrations of platelet factor 4, ss-thromboglobulin, and poly morpho-nuclear (PMN)-elastase were lowest in group C, whereas group C also had the highest concentrations of thrombin-antithrombin complex (P<0.018-0.001). Blood loss within the first 12 h postoperatively was 457 +/- 204 mL in group A, 431 +/- 178 mL in group B, and 382 +/- 188 mL in group C (P<0.01). Complication rates and 30-day mortality did not differ between study groups. CONCLUSION: The combined use of heparin-coated circuits and low dose systemic heparinization is able to reduce early postoperative blood loss without enhancing the risk of complications. PMID- 18431351 TI - Concomitant carotid endarterectomy and coronary artery bypass grafting versus staged carotid stenting followed by coronary artery bypass grafting. AB - AIM: Significant carotid stenosis (>or=70%) in patients undergoing coronary artery bypass grafting (CABG) can increase the risk of perioperative cerebral vascular accident (CVA). In this study, we compared the results of two common operative strategies: concomitant carotid endarterectomy and CABG versus carotid stenting and CABG. METHODS: This cohort study was conducted from January 2001 to September 2006. Significant carotid artery stenosis was detected in patients who were candidates for CABG at the Tehran Heart Center. The stenosis was detected by carotid Doppler screening and was confirmed by magnetic resonance angiography. Reluctant patients or those with previous major CVA, significant bilateral carotid stenosis and intracranial lesions were excluded. Patients were divided into 2 groups. Group A underwent concomitant carotid endarterectomy and CABG (n=19), while carotid stenting and CABG were done in group B (n=28). RESULTS: The mean age in group A was 67.37+/-7.09 years and 65.57+/-8.13 years in group B. The mean hospital stay (days) was 18.68+/-7.95 in group A and 26.35+/-77.04 in group B (P=0.01). The median charge was dollars 252.79 in group A and dollars 2206.66 in group B (P <0.0001). There was a significant difference in frequency of hypotension and bradycardia between the 2 groups (P <0.05). There were 2 cases of in-hospital mortality in each group (10.5% and 7.1%, respectively). Two postoperative strokes occurred in group A and 3 in group B (10.5% and 10.7%, respectively). CONCLUSION: Concomitant carotid endarterectomy and CABG is as safe as carotid stenting and CABG, with fewer neurologic events and less hypotension, bradycardia, cost and shorter hospital stay. PMID- 18431352 TI - Unilateral diaphragm paralysis: etiology, impact, and natural history. AB - AIM: The etiology, clinical impact, natural history and best therapy of unilateral diaphragm paralysis (UDP) are incompletely understood. This condition is not amenable to pacing, which requires an intact phrenic nerve. METHODS: Clinical records of patients with UDP referred to our diaphragm center were reviewed. RESULTS: Thirty-six patients (28 male, 8 female) aged 1 month to 78 years (mean 47.8 years) with UDP evaluated from 1983 to February 2007 were reviewed. Etiology was postsurgical in 13 (36%), tumor (with surgery or radiation therapy) in 7 (19%), idiopathic in 6 (17%), trauma (motor vehicle accident or head injury) in 5 (14%), polio in 3 (8%), and viral in 2 (6%) patients. 28 patients (78%) were symptomatic; 8 (22%) carried a diagnosis of coexisting chronic obstructive pulmonary disease. Mean duration of paralysis was 57.9 months (range up to 261 months). The left diaphragm was involved in 23 cases (64%) and the right in 13 (36%). Mean forced expiratory volume (FEV1) was 1 915 mL (61.3% of predicted) and mean forced vital capacity (FVC) was 2 432 mL (62.9% of predicted). Mean pO2 was 69.9 mmHg (range 49 to 124), indicating considerable shunting through underventilated lung. Pulmonary infection affected 3 patients (8.4%). Diaphragm function returned in 17% of patients (mainly children) at mean of 10.3 months. Four incapacitated patients (11 %) were treated surgically, with resection of the hemi-diaphragm. Surgical exploration revealed neurogenic atrophy of the diaphragm muscle. All 4 resected patients showed clinical, oxymetric, and spirometric improvement. CONCLUSION: The conclusion is drawn that: 1) UDP may be traumatic, tumor-related, iatrogenic, or idiopathic; 2) UDP decreases pO(2) substantially and breathing capacity by more than 1/3; 3) spontaneous recovery is possible; 4) UDP is not intrinsically lethal; 5) occasional patients are incapacited; 6) diaphragm resection produces clinical improvement via lower lobe re-expansion; 7) the incapacity incurred by UDP is mild compared to the clinical spectrum of bilateral diaphragm paralysis. PMID- 18431353 TI - Aortic valve stenosis and regurgitation: an overview of management. AB - There are few cardiovascular disease entities that have as an impressive contribution to improved quality and duration of life as aortic valve disease correction. Although aortic valve stenosis is fairly common, particularly in elderly patients, between one third and two thirds go untreated, despite operative survival rates with the minimal invasive J incision approach of 99%. For those patients with high risk co-morbidity or technical inoperable reasons, the percutaneous transfemoral or transapical approaches are now available with good initial RESULTS: For younger patients with tricuspid valves or bicuspid valves and aortic regurgitation, aortic valve repair should be the first choice procedure and with careful selection of the appropriate approach based on analysis of the patients CLASS (Commissure, Leaflet, Annulus, Sinotubular junction, and Sinuses) anatomy. Excellent results can be obtained with better than 90% freedom from reoperation at 10 years. The scope of procedures for aortic valve disease has increased markedly over the last 10 years and patients and cardiovascular physicians need to be aware of these newer options. PMID- 18431354 TI - Aneurysmal expansion of the aortic patch after thoracoabdominal aortic aneurysm repair. PMID- 18431355 TI - Tracheal stenosis following intubation: is it an early problem? PMID- 18431356 TI - Dendritic cell behaviour in vivo: lessons learned from intravital two-photon microscopy. AB - Dendritic cells (DC) are central regulators of immune responses. Their functional characterization has, thus far, mainly relied on the analysis of ex vivo isolated cells or immunohistology, which provides information in a static manner. While these approaches have enabled an excellent understanding of the role of DC in antigen uptake, processing and presentation, there has been a clear need to investigate the behaviour of DC in the context of intact tissues in real time. This demand has recently been met by the availability of intravital two-photon microscopy, which allows for the visualization of single cells deep within intact organs over time. Thus, during the past few years, exciting new data have been generated as to how DC behave within secondary lymphoid and peripheral tissues both under homoeostatic and inflammatory conditions. Here, we will review what two-photon microscopy studies have taught us about the migration of DC in the interstitial space as well as their interactions with adaptive immune cells. PMID- 18431357 TI - A case of a lower pole renal calculus managed with percutaneous nephrolithotomy. AB - BACKGROUND: A 50-year-old man presented to an outpatient urology clinic with left sided flank pain. He had received treatment for a left renal pelvis calculus 5 years earlier; shock wave lithotripsy had been unsuccessful and ureteroscopic laser lithotripsy had left the patient with a 2 mm residual stone fragment. The fragment was deemed clinically insignificant, and the patient remained asymptomatic without any further follow-up. INVESTIGATIONS: Physical examination, CT of the abdomen and pelvis, and complete metabolic evaluation. DIAGNOSIS: Left lower pole renal calculus and idiopathic hypercalciuria. MANAGEMENT: The patient was counseled about his treatment options, and he elected to undergo percutaneous nephrolithotomy. After a single-stage procedure, CT confirmed a stone-free state. The patient was administered thiazide therapy and dietary counseling to treat his hypercalciuria. This condition resolved, and the patient remained stone-free 1 year after treatment. PMID- 18431358 TI - Can exercise ameliorate the increased risk of cardiovascular disease and diabetes associated with ADT? PMID- 18431360 TI - Expressed anti-HBV primary microRNA shuttles inhibit viral replication efficiently in vitro and in vivo. AB - The use of RNA interference (RNAi) to inhibit gene expression is potentially applicable in the treatment of viral infections such as hepatitis B virus (HBV) persistence. Although efficient HBV gene silencing by short hairpin RNA (shRNA) expressed from RNA polymerase (Pol) III promoters has been reported, constitutive high-level transcription may cause harmful side effects. Here, we report an approach that allows the use of a Pol II promoter to improve transcription regulation of expressed RNAi effecters. Pol II [cytomegalovirus (CMV)] or Pol III (U6) promoter cassettes that transcribe anti-HBV primary microRNA (pri-miR)-122 and pri-miR-31 shuttles were generated. In cultured cells both types of pri-miR like sequences effected knockdown of markers of viral replication (>80%) and were processed to form intended 21-nucleotide guides. The concentration of CMV expressed miRs was approximately 85-fold lower than the U6 shRNA-derived guide RNA. When cells were co-transfected with pri-miR expression cassettes, attenuation of independent RNAi-mediated gene silencing was not observed, which is in contrast to the action of U6 shRNA expression cassettes. The efficacy of the anti-HBV pri-miR shuttles in vivo was verified using the murine hydrodynamic injection model. Employing Pol II-expressed pri-miR mimics may be useful in the treatment of HBV infection, and potentially also for generic application in RNAi based therapy. PMID- 18431359 TI - Treg depletion-enhanced IL-2 treatment facilitates therapy of established tumors using systemically delivered oncolytic virus. AB - There are several roadblocks that hinder systemic delivery of oncolytic viruses to the sites of metastatic disease. These include the tumor vasculature, which provides a physical barrier to tumor-specific virus extravasation. Although interleukin-2 (IL-2) has been used in antitumor therapy, it is associated with endothelial cell injury, leading to vascular leak syndrome (VLS). Here, we demonstrate that IL-2-mediated VLS, accentuated by depletion of regulatory T cells (Treg), facilitates localization of intravenously (i.v.) delivered oncolytic virus into established tumors in immune-competent mice. IL-2, in association with Treg depletion, generates "hyperactivated" natural killer (NK) cells, possessing antitumor activity and secreting factors that facilitate virus spread/replication throughout the tumor by disrupting the tumor architecture. As a result, the combination of Treg depletion/IL-2 and systemic oncolytic virotherapy was found to be significantly more therapeutic against established disease than either treatment alone. These data demonstrate that it is possible to combine biological therapy with oncolytic virotherapy to generate systemic therapy against established tumors. PMID- 18431361 TI - Regulated insulin delivery from human epidermal cells reverses hyperglycemia. AB - Alternative insulin therapies are being sought that will provide euglycemic control for people with diabetes mellitus. The epidermis is a self-renewing tissue that is easily accessible and can provide large numbers of autologous cells that can be used for generating insulin-secreting skin substitutes. Lentiviral vectors have been engineered to produce a fusion protein between the furin-cleavable proinsulin and the self-dimerization mutant of FK506-binding protein to yield bioactive insulin in keratinocytes; this insulin is released as a response to exogenous administration of a small organic molecule, rapamycin. The engineered keratinocytes retained normal morphology and grew in a manner similar to lentiviral-treated control cells. Epidermal keratinocytes in culture and in stratified bioengineered epidermis released insulin within 30 minutes after addition of rapamycin, and secretion slowed or stopped within 2-3 hours after removal of the inducing agent. When the cells were implanted into athymic mice that had been rendered diabetic with streptozotocin (STZ), insulin was detected in the plasma within 1 hour after addition of rapamycin. Concomitantly, serum glucose decreased to normal levels even in diabetic animals with severe hyperglycemia. Repeated rapamycin administration yielded similar results. These experiments provide proof-of-concept that insulin released from the skin in a regulatable manner can reverse hyperglycemia. PMID- 18431362 TI - Enhancing the T-cell stimulatory capacity of human dendritic cells by co electroporation with CD40L, CD70 and constitutively active TLR4 encoding mRNA. AB - The effectiveness of the dendritic cell (DC) vaccination protocols that are currently in use could be improved by providing the DCs with a more potent maturation signal. We therefore investigated whether the T-cell stimulatory capacity of human monocyte-derived DCs could be increased by co-electroporation with different combinations of CD40L, CD70, and constitutively active toll-like receptor 4 (caTLR4) encoding mRNA. We show that immature DCs electroporated with CD40L and/or caTLR4 mRNA, but not those electroporated with CD70 mRNA, acquire a mature phenotype along with an enhanced secretion of several cytokines/chemokines. Moreover, these DCs are very potent in inducing naive CD4(+) T cells to differentiate into interferon-gamma (IFN-gamma)-secreting type 1 T helper (Th1) cells. Further, we assessed the capacity of the electroporated DCs to activate naive HLA-A2-restricted MelanA-specific CD8(+) T cells without the addition of any exogenous cytokines. When all three molecules were combined, a >500-fold increase in MelanA-specific CD8(+) T cells was observed when compared with immature DCs, and a >200-fold increase when compared with cytokine cocktail matured DCs. In correlation, we found a marked increase in cytolytic and IFN gamma/tumor necrosis factor-alpha (TNF-alpha) secreting CD8(+) T cells. Our data indicate that immature DCs genetically modified to express stimulating molecules can induce tumor antigen-specific T cells in vitro and could prove to be a significant improvement over DCs matured with the methods currently in use. PMID- 18431363 TI - Therapeutic effect of recombinant adenovirus encoding interferon-gamma in a murine model of progressive pulmonary tuberculosis. AB - We constructed recombinant adenoviruses encoding murine interferon-gamma (AdIFNgamma) and tested its therapeutic efficiency in a well characterized model of progressive pulmonary tuberculosis (TB) in Balb/c mice, infected through the trachea with the laboratory drug-susceptible H37Rv strain or multidrug-resistant (MDR) clinical isolate. When the disease was in a late phase, 2 months after infection, we administered by intratracheal cannulation a single dose [1.7 x 10(9) plaque forming units (pfu)] of AdIFNgamma or the control adenovirus. Groups of mice were killed at different time-points and the lungs were examined to determine bacilli colony forming units (CFU), cytokine/chemokine gene expression, and CD4/CD8 subpopulations, and also subjected to automated histomorphometry. In comparison with the control group, after 2 weeks of treatment and during the next 6 months, AdIFNgamma-treated animals infected with either the H37Rv strain or the MDR strain showed significantly lower bacilli loads and tissue damage (pneumonia), higher expressions of IFN-gamma, tumor necrosis factor (TNF), and inducible nitric oxide synthase (iNOS), and bigger granulomas. When compared with the results from conventional chemotherapy or AdIFNgamma treatment alone, the combined treatment with AdIFNgamma plus conventional chemotherapy shortened the time taken for reduction of bacillary load. This shows that gene therapy with AdIFNgamma efficiently reconstituted the protective immune response and controlled the progress of pulmonary TB produced by MDR or non-MDR strains. PMID- 18431364 TI - Adult bone marrow-derived cells do not acquire functional attributes of cardiomyocytes when transplanted into peri-infarct myocardium. AB - The cardiomyogenic potential of adult bone marrow (BM) cells after being directly transplanted into the ischemically injured heart remains a controversial issue. In this study, we investigated the ability of transplanted BM cells to develop intracellular calcium ([Ca(2+)](i)) transients in response to membrane depolarization in situ. Low-density mononuclear (LDM) BM cells, c-kit-enriched (c kit(enr)) BM cells, and highly enriched lin(-) c-kit(+) BM cells were obtained from adult transgenic mice ubiquitously expressing enhanced green fluorescent protein (EGFP), and injected into peri-infarct myocardiums of nontransgenic mice. After 9-10 days the mice were killed, and the hearts were removed, perfused in Langendorff mode, loaded with the calcium-sensitive fluorophore rhod-2, and subjected to two-photon laser scanning fluorescence microscopy (TPLSM) to monitor action potential-induced [Ca(2+)](i) transients in EGFP-expressing donor-derived cells and non-expressing host cardiomyocytes. Whereas spontaneous and electrically evoked [Ca(2+)](i) transients were found to occur synchronously in host cardiomyocytes along the graft-host border and in areas remote from the infarct, they were absent in all of the >3,000 imaged BM-derived cells that were located in clusters throughout the infarct scar or peri-infarct zone. We conclude that engrafted BM-derived cells lack attributes of functioning cardiomyocytes, calling into question the concept that adult BM cells can give rise to substantive cardiomyocyte regeneration within the infarcted heart. PMID- 18431365 TI - Relationships between leptin and C-reactive protein with cardiovascular disease in the adult general population. AB - BACKGROUND: Leptin could be a key regulator of C-reactive protein (CRP) levels, which serve as a marker of systemic inflammation. Both leptin and CRP are predictors of cardiovascular disease (CVD). However, the interactions between leptin and CRP, and their association with CVD, remain unclear. We therefore studied them in a large, multiethnic population. METHODS: We analyzed leptin and CRP levels, anthropometric variables and cardiovascular risk factor data from 6,251 participants from the Third National Health and Nutrition Examination Survey (NHANES III). Logistic regression was used to estimate the association between leptin, CRP and CVD (defined as history of myocardial infarction or stroke). Receiver operating characteristic curves were created to study the additional value of leptin and CRP for the association with CVD. RESULTS: The mean age was 44.4 +/- 0.21 years (52.5% women). After adjustment for age, race, dyslipidemia, hypertension, diabetes, smoking, obesity and CRP, high levels of leptin were significantly associated with CVD in men (odds ratio 2.47, 95% CI 1.19-5.19) and in women (odds ratio 3.30, 95% CI 1.47-7.99). After adjustment for leptin, CRP was not associated with CVD. There was a significant correlation between levels of leptin and CRP (Spearman correlation rho = 0.22 in men and rho = 0.32 in women, both P < 0.0001). The area under the curve, representing the association between cardiovascular risk factors and CVD, increased after the addition of high levels of both leptin and CRP together. CONCLUSION: High leptin levels are independently associated with CVD even after adjustment for CRP; elevated CRP levels are not associated with CVD after adjustment for leptin. However, increased concentrations of both leptin and CRP confer the highest risk for CVD. PMID- 18431366 TI - Etiology of cognitive change after CABG surgery: more than just the pump? PMID- 18431367 TI - Spotlight on HDL-raising therapies: insights from the torcetrapib trials. AB - Subnormal levels of HDL cholesterol constitute a major cardiovascular risk factor. Inhibitors of cholesteryl ester transfer protein (CETP) are presently the most potent HDL-raising agents. Torcetrapib was the first CETP inhibitor to enter a large-scale, prospective, placebo-controlled interventional trial, which was prematurely terminated in December 2006 because of excess cardiovascular and noncardiovascular mortality in the active treatment group. Therapy with torcetrapib was associated with considerable increases in aldosterone level and blood pressure and changes in serum electrolytes indicative of mineralocorticoid excess. These findings indicate that torcetrapib has off-target toxic effects unrelated to HDL raising that involve the activation of mineralocorticoid receptors by aldosterone and result in the induction of hypertension. In contrast with torcetrapib, other CETP inhibitors such as JTT-705 and MK-825 do not increase blood pressure in humans, an observation which discounts a class effect. The available data do not, however, exclude potential adverse effects of CETP inhibition such as the generation of HDL particles that have deficient biological activities and a deleterious impact on reverse cholesterol transport and steroid metabolism. Normalization of both defective HDL function and diminished HDL levels should, therefore, be the focus of pharmacological HDL raising in future studies. PMID- 18431368 TI - Anacetrapib: new hope for cholesteryl ester transfer protein inhibitors in the treatment of dyslipidemia. PMID- 18431369 TI - Do all drugs and treatments for rheumatoid arthritis have the same efficacy? PMID- 18431370 TI - Are modified-release corticosteroids good therapeutic options for patients with RA? AB - Glucocorticoids have good efficacy as anti-inflammatory agents but are associated with adverse effects. Buttgereit et al. have compared the efficacy of modified release prednisone with that of standard, immediate-release prednisone for the treatment of rheumatoid arthritis. The modified-release tablets are designed to liberate the glucocorticoid approximately 4 h after ingestion, and can be taken at bedtime so that the prednisone release coincides with peaks in endogenous cortisol levels and symptoms of disease in the early hours of the morning. The authors of this study hypothesized that the modified release would improve the benefit-risk ratio of this glucocorticoid. The 12-week, multicenter, double blind, randomized controlled trial included 251 patients who completed the study. In patients with active rheumatoid arthritis, prescription of the modified release tablet for night-time ingestion resulted in a mean 29.2 min shorter duration of morning stiffness than in patients who received standard immediate release prednisone tablets in the morning, and a total 44.0 min reduction in morning-stiffness duration at 12 weeks compared with baseline. The safety profiles of the modified-release and immediate-release forms of prednisone were similar. PMID- 18431371 TI - Technology insight: noninvasive assessment of bone strength in osteoporosis. AB - Fractures that result from osteoporosis are an enormous and growing concern for public health systems; as the population ages, the number of fractures worldwide will double or triple in the next 50 years. The ability of a bone to resist fracture depends not only on the amount of bone present, but also on the spatial distribution of the bone mass, the cortical and trabecular microarchitecture, and the intrinsic properties of the materials that comprise the bone. Although low bone mineral density is one of the strongest risk factors for fracture, a number of clinical studies have demonstrated the limitations of using measurements of areal bone mineral density by dual-energy X-ray absorptiometry to assess fracture risk and to monitor responses to therapy. As a result, new, noninvasive imaging techniques that are capable of assessing various components of bone strength are being developed. These techniques include three-dimensional assessments of bone density, geometry and microarchitecture, as well as integrated measurements of bone strength by engineering analyses. Although they show strong potential, further development and validation of these techniques is needed to define their role in the clinical management of individuals with osteoporosis. PMID- 18431372 TI - Long-term effects of pegvisomant in patients with acromegaly. AB - Despite improved surgical and radiotherapy techniques and the development of long acting somatostatin analogs, some patients with acromegaly cannot attain biochemical remission. As a consequence they continue to endure debilitating symptoms and mortality remains high. Pegvisomant, a recombinant growth-hormone receptor antagonist, suppresses production of insulin-like growth factor I. Since the introduction of this drug several years ago, long-term studies involving hundreds of patients have established efficacy of more than 85%. Raised transaminase concentrations have, however, been reported as a side effect of therapy, albeit an infrequent one. In addition, increases in tumor volume have been reported in several cases. In this Review we present the long-term data that have been gathered on pegvisomant therapy, discuss the related risks and benefits, and frame a potential therapeutic approach. PMID- 18431373 TI - Is vagal-sparing esophagectomy ideal for Barrett's esophagus with intramucosal adenocarcinoma or high-grade dysplasia? PMID- 18431374 TI - Technology insight: metabonomics in gastroenterology-basic principles and potential clinical applications. AB - Metabonomics-the study of metabolic changes in an integrated biologic system-is an emerging field. This discipline joins the other 'omics' (genomics, transcriptomics and proteomics) to give rise to a comprehensive, systems-biology approach to the evaluation of holistic in vivo function. Metabonomics, especially when based on nuclear magnetic resonance spectroscopy, has the potential to identify biomarkers and prognostic factors, enhance clinical diagnosis, and expand hypothesis generation. As a consequence, the use of metabonomics has been extensively explored in the past decade, and applied successfully to the study of human diseases, toxicology, microbes, nutrition, and plant biology. This Review introduces the basic principles of nuclear magnetic resonance spectroscopy and commonly used tools for multivariate data analysis, before considering the applications and future potential of metabonomics in basic and clinical research, with emphasis on applications in the field of gastroenterology. PMID- 18431375 TI - Does prophylactic cranial irradiation reduce the incidence of brain metastases in extensive small-cell lung cancer? PMID- 18431376 TI - Rapamycin treatment for a child with germline PTEN mutation. AB - BACKGROUND: A 9-month-old boy with Proteus syndrome and a de novo germline mutation in the tumor suppressor PTEN was referred to a specialist centre for management. Over the first years of life, the patient developed life-threatening respiratory dysfunction and malnutrition because of progressive growth of hamartomas affecting the chest, mediastinum, abdomen and pelvis. INVESTIGATIONS: Physical examination, CT scans of the mediastinum, pelvis and abdomen, measurement of serum insulin-like growth factor binding protein-2, and investigation of the effect of the PTEN mutation on phosphatidylinositol 3 kinase/mammalian target of rapamycin signaling in an in vitro cell model. DIAGNOSIS: PTEN hamartoma tumor syndrome, specifically Proteus syndrome. MANAGEMENT: Oral rapamycin. PMID- 18431377 TI - Mechanisms of Disease: cancer stem cells--targeting the evil twin. AB - Classical antineoplastic treatments such as chemotherapy or radiation can efficiently eradicate the majority of proliferating and genetically unstable malignant cells within neoplastic lesions. There is increasing evidence, however, that these regimens frequently fail to eliminate a minor subpopulation of resistant tumor cells that have distinct features of somatic stem cells. These serve as a reservoir for disease recurrence, and are the origin of metastatic growth. These so-called cancer stem cells (CSCs) or cancer-initiating cells represent often a rare, highly self-renewing population within the tumor mass, which is thought to be the only one required for both initiation and maintenance of disease. Tumor-cell populations enriched for CSC activity were originally identified in leukemias, but have now also been uncovered in a number of solid cancers. Their marked resistance towards classical antitumor regimens is mediated by the combination of several critical features, including relative dormancy, efficient DNA repair, high expression of multidrug-resistance-type membrane transporters and protection by a hypoxic niche environment. We review the concept of CSCs with particular emphasis on the mechanism of therapy resistance, and discuss potential future therapeutic interventions with the goal of specifically eliminating CSCs in a clinical setting. PMID- 18431378 TI - Recommendations and guidelines for the diagnosis and treatment of Fabry nephropathy in adults. AB - Progressive loss of kidney function complicates Fabry disease, an X-linked lysosomal storage disorder that arises from deficiency of alpha-galactosidase activity. Heterozygous females with Fabry disease can be as severely affected as hemizygous males, who have the classic form of the disease. Enzyme-replacement therapy with recombinant human alpha-galactosidase clears the glycosphingolipid globotriaosylceramide from kidney cells, and can stabilize renal function in adults with mild to moderate Fabry nephropathy. However, adults with more advanced nephropathy and overt proteinuria do not respond as well. For these patients, antiproteinuric therapy given in conjunction with enzyme-replacement therapy might prevent further decline in kidney function. In this Review, we propose guidelines and recommendations for the diagnosis and management of Fabry nephropathy in adults, based on published data and on the consensus of opinion of participants in the 7(th) International Fabry Nephropathy Roundtable in 2007. These organ-specific guidelines could be easier to implement than general guidelines, provided they are used in the context of an overall multisystem care approach. PMID- 18431379 TI - 'Noisy patients'--can signal detection theory help? AB - Signal detection theory tests an observer's ability to discriminate between signal and noise. Deciding whether or not a patient's symptoms warrant further investigation or treatment is an example of this task in the clinical setting. Noise can exist within the observer--for example, in the brain of a tired or inexperienced doctor--or can arise from an external source such as the patient. Patients can produce external noise by giving numerous unrelated presenting complaints, providing overly detailed accounts of their symptoms, or simply talking too quickly. The more noise that is present, the harder the signal (such as a new disease or a notable change in an old condition) is to detect. Patients in the neurology clinic seem to be 'noisier' than average, perhaps owing to the long duration of their condition in many cases and the relatively high proportion of patients with medically unexplained symptoms. The ability to interpret such 'noisy' histories often underpins the neurological diagnosis. This Review aims to promote the relevance of signal detection theory to the overworked neurologist on the ward or in the clinic and explores strategies to reduce the noise generated both within the brain of the doctor and by patients. PMID- 18431380 TI - Noninvasive mechanical ventilation in patients with myasthenic crisis. PMID- 18431381 TI - Facial expressions, pain and nociception--are they related? PMID- 18431387 TI - Animal number justification. Stronger rationales encouraged. PMID- 18431388 TI - Animal number justification. PMID- 18431389 TI - Animal number justification. Statistics should be used. PMID- 18431390 TI - Animal number justification. Precise justification is needed. PMID- 18431391 TI - Perineal swelling in a mouse. Diagnosis: imperforate vagina with secondary mucometra. PMID- 18431393 TI - Vaginoscopic urinary catheter placement in female dogs. AB - Urinary catheter placement in dogs may be necessary for certain surgical procedures or for treatment of certain conditions. This column describes the vaginoscopic placement of an indwelling female urinary catheter and possible complications of catheter placement. PMID- 18431394 TI - Toxicology and pathology considerations for the design of juvenile animal studies. AB - Although exposure to drugs or toxicants can affect children and adults very differently, many compounds lack specific safety information for children. Studies in juvenile animals can help researchers assess pediatric patients' potential response to certain chemicals. Juvenile studies are highly sensitive to animal age, sex and species and must be planned with care to prevent misinterpretation of experimental data. The author reviews considerations for the design of these studies, focusing on toxicological and pathological aspects. PMID- 18431395 TI - Comparison of methods for detection of pinworms in mice and rats. AB - Though pinworm infestation remains common in laboratory rodent colonies, there is little information regarding current practices for pinworm detection and their relative efficacy. The authors surveyed research institutions to find out the prevalence of pinworm infestations and the detection methods they used. They also tested mice and rats from colonies that were known to be infested with Syphacia sp. and compared the following detection methods: perianal tape test, fecal flotation, fecal concentration, cecal content examination, cecal flotation and histological examination. Though the different methods yielded comparable efficacy overall, the authors recommend using more than one type of test to increase detection potential. PMID- 18431396 TI - Report of the 2006 RSPCA/UFAW Rodent Welfare Group meeting. AB - The RSPCA/UFAW Rodent Welfare Group holds a one-day meeting every autumn to discuss current welfare research and to exchange views on rodent welfare issues. A key aim of the group is to encourage people to think about the lifetime experience of laboratory rodents, ensuring that every potential influence on their well-being has been reviewed and refined. Speakers at the 2006 meeting presented preliminary findings of ongoing studies and discussed regulatory updates. Topics included the housing and husbandry of mice and rats, refining the use of rodents in asthma research, good practice for the euthanasia of rodents using carbon dioxide and achieving reduction by sharing genetically modified mice. PMID- 18431397 TI - Fire safety recommendations for administration of isoflurane anesthesia in oxygen. AB - A researcher at the author's facility was carrying out a routine surgical procedure in a mouse that was anesthetized with vaporized isoflurane. When the researcher brought an active cauterizer close to the mouse, a flame erupted from the anesthesia nosecone. An investigation concluded that the fire was ignited when the cauterizer came into contact with the oxygen-enriched atmosphere that was streaming from the anesthetic equipment. The author presents recommendations for preventing similar incidents in the future. PMID- 18431399 TI - Mechanisms of asymmetric cell division: flies and worms pave the way. AB - Asymmetric cell division is fundamental for generating diversity in multicellular organisms. The mechanisms that govern asymmetric cell division are increasingly well understood, owing notably to studies that were conducted in Drosophila melanogaster and Caenorhabditis elegans. Lessons learned from these two model organisms also apply to cells that divide asymmetrically in other metazoans, such as self-renewing stem cells in mammals. PMID- 18431400 TI - Transcriptional control of human p53-regulated genes. AB - The p53 protein regulates the transcription of many different genes in response to a wide variety of stress signals. Following DNA damage, p53 regulates key processes, including DNA repair, cell-cycle arrest, senescence and apoptosis, in order to suppress cancer. This Analysis article provides an overview of the current knowledge of p53-regulated genes in these pathways and others, and the mechanisms of their regulation. In addition, we present the most comprehensive list so far of human p53-regulated genes and their experimentally validated, functional binding sites that confer p53 regulation. PMID- 18431401 TI - Membrane nanotubes: dynamic long-distance connections between animal cells. AB - Membrane nanotubes are transient long-distance connections between cells that can facilitate intercellular communication (for example, by trafficking vesicles or transmitting calcium-mediated signals), but they can also contribute to pathologies (for example, by directing the spread of viruses). Recent data have revealed considerable heterogeneity in their structures, processes of formation and functional properties, in part dependent on the cell types involved. Despite recent progress in this young research field, further research is sorely needed. PMID- 18431402 TI - Use of acute hyperphenylalaninemia in rhesus monkeys to examine sensitivity and stability of the L-[1-11C]leucine method for measurement of regional rates of cerebral protein synthesis with PET. AB - We have previously shown by direct comparison with autoradiographic and biochemical measurements that the L-[1-(11)C]leucine positron emission tomography method provides accurate determinations of regional rates of cerebral protein synthesis (rCPS) and the fraction (lambda) of unlabeled leucine in the precursor pool for protein synthesis derived from arterial plasma. In this study, we examine sensitivity of the method to detect changes in lambda and stability of the method to measure rCPS in the face of these changes. We studied four isoflurane-anesthetized monkeys dynamically scanned with the high resolution research tomograph under control and mild hyperphenylalaninemic conditions. Hyperphenylalaninemia was produced by an infusion of phenylalanine that increased plasma phenylalanine concentrations three- to five-fold. In phenylalanine-infused monkeys, plasma leucine concentrations remained relatively constant, but values of lambda were statistically significantly decreased by 11% to 15%; rCPS was unaffected. Effects on lambda are consistent with competitive inhibition of leucine transport by increased plasma phenylalanine. The effect on lambda shows that competition for the transporter results in a reduction in the fraction of leucine in the precursor pool for protein synthesis coming from plasma. Even under these hyperphenylalaninemic conditions, rCPS remains unchanged due to the compensating increased contribution of leucine from protein degradation to the precursor pool. PMID- 18431403 TI - Revisiting horizontal transfer of transposable elements in Drosophila. AB - Horizontal transfer (HT), defined as the transfer of genetic material between species, is considered to be an essential step in the 'life cycle' of transposable elements. We present a broad overview of suspected cases of HT of transposable elements in Drosophila. Hundred-one putative events of HT have been proposed in Drosophila for 21 different elements (5.0% refer to non-long terminal repeat (LTR) retrotransposons, 42.6% to LTR retrotransposons and 52.4% to DNA transposons). We discuss the methods used to infer HT, their limits and the putative vectors of transposable elements. We outline all the alternative hypotheses and ask how we can be almost certain that phylogenetic inconsistencies are due to HT. PMID- 18431405 TI - Family communication between children and their parents about inherited genetic conditions: a meta-synthesis of the research. AB - In families affected by an inherited genetic condition, parents face a difficult task of having to communicate genetic risk information to their children. A systematic review of all major health and medical research databases was undertaken using current guidelines to identify original relevant research papers from 1980 to 2007, which explore the issues surrounding parents and their children's communication about inherited genetic risk. A total of 9698 abstracts were found of which 158 research papers were reviewed as potentially relevant. A final 17 papers were identified which met predefined inclusion and exclusion criteria. Using a meta-ethnographic approach, all identified studies' findings were analysed as primary data sources by three researchers, who independently identified the key concepts. A high level of congruence emerged between researchers, and agreed concepts were used to examine similarities and differences between papers. The findings informed the development of a narrative framework exploring the issues that related to parents' explanations of inherited genetic risk to their children, the reasons for sharing information, children's understanding of parents' explanations, the emotions evoked for family members and the support and guidance received from health professionals. Providing information, checking understanding, and explaining and managing the emotional feelings that arise were integral to supporting children's coping with genetic risk information. However, many parents struggled with one or more of these components and required more support specific to the child's developmental stage, and family members' transition of readjustment to the impact of the genetic condition. PMID- 18431404 TI - A cAMP-specific phosphodiesterase (PDE8B) that is mutated in adrenal hyperplasia is expressed widely in human and mouse tissues: a novel PDE8B isoform in human adrenal cortex. AB - Bilateral adrenocortical hyperplasia (BAH) is the second most common cause of corticotropin-independent Cushing syndrome (CS). Genetic forms of BAH have been associated with complex syndromes such as Carney Complex and McCune-Albright syndrome or may present as isolated micronodular adrenocortical disease (iMAD) usually in children and young adults with CS. A genome-wide association study identified inactivating phosphodiesterase (PDE) 11A (PDE11A)-sequencing defects as low-penetrance predisposing factors for iMAD and related abnormalities; we also described a mutation (c.914A > C/H305P) in cyclic AMP (cAMP)-specific PDE8B, in a patient with iMAD. In this study we further characterize this mutation; we also found a novel PDE8B isoform that is highly expressed in the adrenal gland. This mutation is shown to significantly affect the ability of the protein to degrade cAMP in vitro. Tumor tissues from patients with iMAD and no mutations in the coding PDE8B sequence or any other related genes (PRKAR1A, PDE11A) showed downregulated PDE8B expression (compared to normal adrenal cortex). Pde8b is detectable in the adrenal gland of newborn mice and is widely expressed in other mouse tissues. We conclude that PDE8B is another PDE gene linked to iMAD; it is a candidate causative gene for other adrenocortical lesions linked to the cAMP signaling pathway and possibly for tumors in other tissues. PMID- 18431407 TI - Simulated drug administration: an emerging tool for teaching clinical pharmacology during anesthesiology training. AB - A thorough understanding of the dose-response relationship is required for optimizing the efficacy of anesthetics while minimizing adverse drug effects. Nowadays, except for the inhaled anesthetics (for which end-tidal concentrations can be measured online), most of the drugs used in clinical anesthesia are administered using standard dosing guidelines, without giving due consideration to their pharmacokinetics and dynamics in guiding their administration. Various studies have found that introducing pharmacokinetics and pharmacodynamics as part of the inputs in clinical anesthesiology could lead to better patient care. With this in mind, it is extremely important that clinicians understand and apply the principles of clinical pharmacology that determine the time course of a drug's disposition and effect. Clinical pharmacology is one of the most challenging topics to teach in anesthesiology. The development of simulators to illustrate the time course of a drug's disposition and effect provides online visualization of pharmacokinetic-pharmacodynamic information during the clinical use of anesthetics. The aim of this review is to discuss the importance of simulation as a clinical pharmacology teaching tool for trainees in anesthesiology. PMID- 18431409 TI - Screening, stem cells, biomarkers and hypercholesterolemia: our first original clinical research. PMID- 18431408 TI - Changes in enoxaparin pharmacokinetics during pregnancy and implications for antithrombotic therapeutic strategy. AB - Enoxaparin is frequently prescribed for pregnant women who are at high risk for thromboembolic complications. We conducted a population pharmacokinetics study with 75 pregnant women and 38 nonpregnant women as controls to evaluate enoxaparin pharmacokinetics during pregnancy and the postpartum period. Clearance of the drug was higher in the pregnant women throughout pregnancy when compared with nonpregnant women (0.78 +/- 0.03 l/h vs. 0.52 +/- 0.03 l/h, respectively P < 0.001) with the stage of the pregnancy having no influence. The volume of distribution was influenced by stage of the pregnancy, characterized by a two step increase, with an initial rise paralleling the woman's increase in body weight during the first two trimesters, followed by an additional increase of 41% during the last 2 months of pregnancy, independent of changes in weight. Using enoxaparin pharmacokinetic parameters to simulate anti-Xa time profiles, we observed that the maintenance of the same doses throughout pregnancy resulted in a progressive reduction in mean and peak anti-Xa activities. We recommend the administration of doses normalized for body weight changes so as to counteract enoxaparin pharmacokinetic changes that accompany various stages of pregnancy. PMID- 18431410 TI - Experimental evolution. PMID- 18431411 TI - [Prioritization on the paper--and in practice]. PMID- 18431412 TI - [Chemicals and acute poisonings]. PMID- 18431413 TI - [Geriatric surgery between reflection and resection]. PMID- 18431414 TI - [Lung cancer without metastases--are all patients with resectable tumor operated?]. AB - BACKGROUND: Only 17% of patients with lung cancer are surgically resected, and the resection rate has not improved despite more attention about the disease. All patients with resectable disease should be offered operation and we wanted to investigate whether this is the case. MATERIAL AND METHODS: We assessed patients that received the diagnosis localized lung cancer in the period 2003-2005 and were not resected according to the Cancer Registry of Norway (n = 322). After exclusion of 40 patients, 282 remained for evaluation. RESULTS: The Cancer Registry of Norway had received clinical reports for 253 patients. Lung physicians had filled in less than half of these, and TNM was registered appropriately in 37% of all reports. Despite that all patients had been categorized in the Registry as having localized lung cancer, 55 patients had advanced disease. Poor lung function, high age and serious comorbidity were contraindications towards surgery for patients with localized disease. Of 282 patients, 258 were inoperable while nine had undergone resection. The remaining 15 cases were thus classified as being operable or possibly operable. INTERPRETATION: The proportion of patients who had not undergone resection and were assessed as operable has decreased. Legally required reporting to the Cancer Registry is still insufficient. Introduction of a specific report form for these patients may facilitate treatment evaluation and thereby treatment and ensure that the decision to not operate is well documented. PMID- 18431415 TI - [Inquiries to the Norwegian Poisons Information Centre on acute exposures to chemicals and chemical products]. AB - BACKGROUND: The Poisons Information Centre receives many inquiries about acute exposures to chemical products. Our aim was to characterise the frequency and severity of such exposures and to raise awareness of chemicals that rarely cause poisoning, but may lead to serious systemic toxicity even in small amounts. MATERIAL AND METHODS: Data were collected from inquiries to the Poisons Information Centre in the period 2004-2006 and from the Product Register on the use of selected chemicals. RESULTS: In 2004-2006, the Poisons Information Centre received 35,802 inquiries regarding acute exposures to chemicals or chemical products. Most of the exposures (72%) were assessed as non-toxic or involving risk of minor poisoning, while only 7.8% were assessed as involving risk of severe poisoning. The substances most frequently involved were cleaning agents, cosmetics/personal care products and hydrocarbon fuels, whereas risk of severe poisoning is related to alkali and hydrocarbon exposures. Poisonings with hydrofluoric acid and glycols/glycol ethers are rare, but the outcome is often severe. As many as 84% of the hydrofluoric acid exposures and 36% of the ingestions of ethylene glycol by children were assessed as involving risk of moderate or severe poisoning. INTERPRETATION: Exposure to chemicals or chemical products is frequent, but rarely leads to severe poisonings. Data from inquiries to the Poisons Information Centre are representative of the chemical exposure conditions. It is important to have detailed product information readily available to assure correct treatment after exposure incidents. PMID- 18431416 TI - [An outbreak of methicillin-resistant Staphylococcus aureus infection in a maternity ward]. AB - BACKGROUND: Staphylococcus aureus is a frequent cause of serious infections. Methicillin-resistant S. aureus (MRSA) are resistant to almost all types of beta lactam antibiotics and therefore represent a substantial medical problem. MATERIAL AND METHOD: In April 2006, the Department of Obstetrics at the Asker and Baerum hospital had an outbreak of MRSA that affected four newborns. The source for the infection was sought among family members, other patients and employees, and eradication was attempted. RESULTS: An employee was identified as the probable infectious source. Subsequent investigation identified 13 individuals infected by the same MRSA clone, which was Panton-Valentine-leukocidin (PVL) positive and therefore clearly a pathogen. 10 of the patients had MRSA disease, with 21 months between the first and the last identified case. For 5 of 13 patients MRSA was still detectable after the first attempt of eradication. For 2 patients, including one of the newborns, eradication has so far been unsuccessful and a third patient has acquired a new abscess after one year. INTERPRETATION: Issues connected to MRSA-screening of close contacts and eradication are resource demanding and require careful consideration of strategy, especially for small children and families with chronic MRSA carriers. Updated detailed national guidelines for MRSA management are needed. PMID- 18431417 TI - [Metformin should not be used by patients with reduced renal function]. PMID- 18431418 TI - [Sick listing--health service or disservice?]. PMID- 18431419 TI - [Labeling of healthy food]. PMID- 18431420 TI - [Children with cochlear implants must be guaranteed adequate training in the Norwegian language]. PMID- 18431421 TI - [Boys with ADHD don't get enough help]. PMID- 18431423 TI - [Weight reduction and diabetes--more knowledge available]. PMID- 18431424 TI - [Take labor pain seriously]. PMID- 18431426 TI - [Better mother after a painful labor?]. PMID- 18431427 TI - [Humans behind global warming? Probably!]. PMID- 18431428 TI - [Humans behind global warming]. PMID- 18431430 TI - [Expert testimony in the Torgersen case was not quality assured]. PMID- 18431439 TI - Furan Decorated Nucleoside Analogues as Fluorescent Probes: synthesis, photophysical evaluation and site-specific incorporation. AB - The synthesis and photophysical evaluation of modified nucleoside analogues in which a five-membered heterocycle (furan, thiophene, oxazole and thiazole) is attached to the 5 position of 2'-deoxyuridine are reported. The furan containing derivative is identified as the most promising responsive nucleoside of this family due to its emission quantum efficiency and degree of sensitivity to its microenvironment. The furan moiety was then attached to the 5 position of 2' deoxycytidine as well as the 8 position of adenosine and guanosine. Photophysical evaluation of these four furan containing nucleoside analogues reveal distinct differences in the absorption, emission and quantum efficiency depending upon the class of nucleoside (pyrimidine or purine). Comparing the photophysical properties of all furan containing nucleosides, identifies the furan thymidine analogue, 5-(fur-2-yl)-2'-deoxyuridine, as the best candidate for use as a responsive fluorescent probe in nucleic acids. 5-(fur-2-yl)-2'-deoxyuridine was then converted to the corresponding phosphoramidite and site specifically incorporated into DNA oligonucleotides with greater than 88% coupling efficiency. Such furan-modified oligonucleotides form stable duplexes upon hybridization to their complementary DNA strands and display favorable fluorescent features. PMID- 18431440 TI - Using an emissive uridine analogue for assembling fluorescent HIV-1 TAR constructs. AB - Emissive nucleoside analogues that are sensitive to their microenvironment can serve as probes for exploring RNA folding and recognition. We have previously described the synthesis of an environmentally sensitive furan-containing uridine and its triphosphate, and have demonstrated that T7 RNA polymerase recognizes this modified ribonucleoside triphosphate as a substrate in in vitro transcription reactions. Here we report the enzymatic preparation of fluorescently tagged HIV-1 TAR constructs and study their interactions with a Tat peptide. Two extreme labeling protocols are examined, where either all native uridine residues are replaced with the corresponding modified fluorescent analogue, or only key residues are site-specifically modified. For the HIV-1 Tat TAR system, labeling all native uridine residues resulted in relatively small changes in emission upon increasing concentrations of the Tat peptide. In contrast, when the two bulge U residues were site-specifically labeled, a reasonable fluorescence response was observed upon Tat titration. The scope and limitations of such fluorescently tagged RNA systems are discussed. PMID- 18431441 TI - Synthesis of HIV-1 Psi-site RNA sequences with site specific incorporation of the fluorescent base analog 2-aminopurine. AB - Fluorescent nucleotide base analogs can serve as sensitive probes of the local structure and chemical environment of the base within a nucleic acid sequence. A significant strength of these base analogs is their similarity in molecular constitution and chemical properties to natural bases. While chemical synthesis has afforded the ability to generate oligonucleotides in good yield with sequence specific incorporation of fluorescent base analogs, this method is limited in practice to the synthesis of relatively small RNAs of less than ~ 80 nucleotides. Since most RNAs of biological interest are greater than 80 nucleotides in length, methods for synthesizing these larger RNAs in good yield, while maintaining the ability to site-specifically incorporate base analogs that allow for fluorescence measurements, could be of broad interest. Here we describe an approach for synthesis of large RNA molecules (>100 nt) that uses T4 RNA ligase to segmentally join a sequence fragment of an RNA, chemically synthesized with a fluorescent base analog, with the remaining unmodified portion of the RNA oligonucleotide, synthesized through in vitro transcription with T7 polymerase. This method is demonstrated through synthesis of packaging sequences (Psi-site) derived from HIV 1 genomic RNA leader sequence (~ 120 nt) with the fluorescent base analog, 2 aminopurine (2-AP), selectively incorporated into the dimerization initiation site (DIS) stem-loop sequence. Using 2-AP fluorescence, RNA conformational changes associated with the formation of non-covalent DIS mediated Psi-site dimers have been analyzed. PMID- 18431442 TI - Use of 2-aminopurine as a fluorescent tool for characterizing antibiotic recognition of the bacterial rRNA A-site. AB - Spectroscopic and calorimetric techniques have been employed to characterize the impact of incorporation of the fluorescent base analog 2-aminopurine into the 1492 or 1493 position of an E. coli rRNA A-site model oligonucleotide, as well as the energetics and dynamics associated with recognition of this A-site model oligomer by aminoglycoside antibiotics. The results of these studies indicate that incorporation of 2AP into either the 1492 or 1493 position does not perturb the structure or stability of the host RNA or its aminoglycoside binding affinity. In addition, the results also highlight drug-induced reduction in the mobilities of the bases at both positions 1492 and 1493 as a potentially key determinant of bactericidal potency. PMID- 18431444 TI - A simplified 4-site economical intradermal post-exposure rabies vaccine regimen: a randomised controlled comparison with standard methods. AB - BACKGROUND: The need for economical rabies post-exposure prophylaxis (PEP) is increasing in developing countries. Implementation of the two currently approved economical intradermal (ID) vaccine regimens is restricted due to confusion over different vaccines, regimens and dosages, lack of confidence in intradermal technique, and pharmaceutical regulations. We therefore compared a simplified 4 site economical PEP regimen with standard methods. METHODS: Two hundred and fifty four volunteers were randomly allocated to a single blind controlled trial. Each received purified vero cell rabies vaccine by one of four PEP regimens: the currently accepted 2-site ID; the 8-site regimen using 0.05 ml per ID site; a new 4-site ID regimen (on day 0, approximately 0.1 ml at 4 ID sites, using the whole 0.5 ml ampoule of vaccine; on day 7, 0.1 ml ID at 2 sites and at one site on days 28 and 90); or the standard 5-dose intramuscular regimen. All ID regimens required the same total amount of vaccine, 60% less than the intramuscular method. Neutralising antibody responses were measured five times over a year in 229 people, for whom complete data were available. FINDINGS: All ID regimens showed similar immunogenicity. The intramuscular regimen gave the lowest geometric mean antibody titres. Using the rapid fluorescent focus inhibition test, some sera had unexpectedly high antibody levels that were not attributable to previous vaccination. The results were confirmed using the fluorescent antibody virus neutralisation method. CONCLUSIONS: This 4-site PEP regimen proved as immunogenic as current regimens, and has the advantages of requiring fewer clinic visits, being more practicable, and having a wider margin of safety, especially in inexperienced hands, than the 2-site regimen. It is more convenient than the 8-site method, and can be used economically with vaccines formulated in 1.0 or 0.5 ml ampoules. The 4-site regimen now meets all requirements of immunogenicity for PEP and can be introduced without further studies. TRIAL REGISTRATION: Controlled-Trials.com ISRCTN 30087513. PMID- 18431445 TI - Impact of environment and social gradient on Leptospira infection in urban slums. AB - BACKGROUND: Leptospirosis has become an urban health problem as slum settlements have expanded worldwide. Efforts to identify interventions for urban leptospirosis have been hampered by the lack of population-based information on Leptospira transmission determinants. The aim of the study was to estimate the prevalence of Leptospira infection and identify risk factors for infection in the urban slum setting. METHODS AND FINDINGS: We performed a community-based survey of 3,171 slum residents from Salvador, Brazil. Leptospira agglutinating antibodies were measured as a marker for prior infection. Poisson regression models evaluated the association between the presence of Leptospira antibodies and environmental attributes obtained from Geographical Information System surveys and indicators of socioeconomic status and exposures for individuals. Overall prevalence of Leptospira antibodies was 15.4% (95% confidence interval [CI], 14.0-16.8). Households of subjects with Leptospira antibodies clustered in squatter areas at the bottom of valleys. The risk of acquiring Leptospira antibodies was associated with household environmental factors such as residence in flood-risk regions with open sewers (prevalence ratio [PR] 1.42, 95% CI 1.14 1.75) and proximity to accumulated refuse (1.43, 1.04-1.88), sighting rats (1.32, 1.10-1.58), and the presence of chickens (1.26, 1.05-1.51). Furthermore, low income and black race (1.25, 1.03-1.50) were independent risk factors. An increase of US$1 per day in per capita household income was associated with an 11% (95% CI 5%-18%) decrease in infection risk. CONCLUSIONS: Deficiencies in the sanitation infrastructure where slum inhabitants reside were found to be environmental sources of Leptospira transmission. Even after controlling for environmental factors, differences in socioeconomic status contributed to the risk of Leptospira infection, indicating that effective prevention of leptospirosis may need to address the social factors that produce unequal health outcomes among slum residents, in addition to improving sanitation. PMID- 18431446 TI - Bayesian mixture model analysis for detecting differentially expressed genes. AB - Control-treatment design is widely used in microarray gene expression experiments. The purpose of such a design is to detect genes that express differentially between the control and the treatment. Many statistical procedures have been developed to detect differentially expressed genes, but all have pros and cons and room is still open for improvement. In this study, we propose a Bayesian mixture model approach to classifying genes into one of three clusters, corresponding to clusters of downregulated, neutral, and upregulated genes, respectively. The Bayesian method is implemented via the Markov chain Monte Carlo (MCMC) algorithm. The cluster means of down- and upregulated genes are sampled from truncated normal distributions whereas the cluster mean of the neutral genes is set to zero. Using simulated data as well as data from a real microarray experiment, we demonstrate that the new method outperforms all methods commonly used in differential expression analysis. PMID- 18431447 TI - Gene tree labeling using nonnegative matrix factorization on biomedical literature. AB - Identifying functional groups of genes is a challenging problem for biological applications. Text mining approaches can be used to build hierarchical clusters or trees from the information in the biological literature. In particular, the nonnegative matrix factorization (NMF) is examined as one approach to label hierarchical trees. A generic labeling algorithm as well as an evaluation technique is proposed, and the effects of different NMF parameters with regard to convergence and labeling accuracy are discussed. The primary goals of this study are to provide a qualitative assessment of the NMF and its various parameters and initialization, to provide an automated way to classify biomedical data, and to provide a method for evaluating labeled data assuming a static input tree. As a byproduct, a method for generating gold standard trees is proposed. PMID- 18431448 TI - 3D wavelet subbands mixing for image denoising. AB - A critical issue in image restoration is the problem of noise removal while keeping the integrity of relevant image information. The method proposed in this paper is a fully automatic 3D blockwise version of the nonlocal (NL) means filter with wavelet subbands mixing. The proposed wavelet subbands mixing is based on a multiresolution approach for improving the quality of image denoising filter. Quantitative validation was carried out on synthetic datasets generated with the BrainWeb simulator. The results show that our NL-means filter with wavelet subbands mixing outperforms the classical implementation of the NL-means filter in terms of denoising quality and computation time. Comparison with wellestablished methods, such as nonlinear diffusion filter and total variation minimization, shows that the proposed NL-means filter produces better denoising results. Finally, qualitative results on real data are presented. PMID- 18431450 TI - Cu(II) and Ni(II) interactions with the terminally blocked hexapeptide Ac-Leu-Ala His-Tyr-Asn-Lys-amide model of histone H2B (80-85). AB - The N- and C-terminal blocked hexapeptide Ac-Leu-Ala-His-Tyr-Asn-Lys-amide (LAHYNK) representing the 80-85 fragment of histone H2B was synthesized and its interactions with Cu(II) and Ni(II) ions were studied by potentiometric, UV-Vis, CD, EPR, and NMR spectroscopic techniques in solution. Our data reveal that the imidazole N(3) nitrogen atom is the primary ligating group for both metal ions. Sequential amide groups deprotonation and subsequent coordination to metal ions indicated an {N(imidazole), 3N(amide)} coordination mode above pH approximately 9, in all cases. In the case of Cu(II)-peptide system, the almost exclusive formation of the predominant species CuL in neutral media accounting for almost 98% of the total metal ion concentration at pH 7.3 strongly indicates that at physiological pH values the sequence -LAHYNK- of histone H2B provides very efficient binding sites for metal ions. The imidazole pyrrole N(1) ionization (but not coordination) was also detected in species CuH(-4)L present in solution above pH approximately 11. PMID- 18431449 TI - Arsenic-based antineoplastic drugs and their mechanisms of action. AB - Arsenic-based compounds have become accepted agents for cancer therapy providing high rates of remission of some cancers such as acute promyelocytic leukemia (APL). The mechanisms by which arsenic-containing compounds kill cells and reasons for selective killing of only certain types of cancer cells such as APLs have recently been delineated. This knowledge was gained in parallel with increasing understanding and awareness of the importance of intracellular redox systems and regulation of the production of reactive oxygen species (ROS) by controlling mitochondrial function. Many of the targets for the arsenic containing compounds are mitochondrial proteins involved in regulating the production of ROS. Inhibition of these proteins by disulfide linkage of vicinal thiol groups often leads to increased production of ROS and induction of apoptotic signalling pathways. Sensitivity or resistance to the actions of arsenic-containing compounds on cancer cells and normal cells depends on the levels of transport systems for their uptake or efflux from the cells as well as their redox defence mechanisms. The exact mechanisms of arsenic toxicity as well as its anticancer properties are likely to be related and these aspects of arsenic metabolism are covered in this review. Greater understanding of the mechanisms of action of arsenic will help determine the risks of human exposure to this chemical. Novel organic arsenic-containing compounds and the lessons learned from studying their selective sensitivity in targeting dividing endothelial cells to inhibit angiogenesis raise the future possibility for designing better targeted antineoplastic arsenic-containing compounds with less toxicity to normal cells. PMID- 18431451 TI - Fast parallel molecular algorithms for DNA-based computation: solving the elliptic curve discrete logarithm problem over GF2. AB - Elliptic curve cryptographic algorithms convert input data to unrecognizable encryption and the unrecognizable data back again into its original decrypted form. The security of this form of encryption hinges on the enormous difficulty that is required to solve the elliptic curve discrete logarithm problem (ECDLP), especially over GF(2(n)), n in Z+. This paper describes an effective method to find solutions to the ECDLP by means of a molecular computer. We propose that this research accomplishment would represent a breakthrough for applied biological computation and this paper demonstrates that in principle this is possible. Three DNA-based algorithms: a parallel adder, a parallel multiplier, and a parallel inverse over GF(2(n)) are described. The biological operation time of all of these algorithms is polynomial with respect to n. Considering this analysis, cryptography using a public key might be less secure. In this respect, a principal contribution of this paper is to provide enhanced evidence of the potential of molecular computing to tackle such ambitious computations. PMID- 18431453 TI - Five novel mutations of the FRMD7 gene in Chinese families with X-linked infantile nystagmus. AB - PURPOSE: Infantile nystagmus (IN) is an inherited disorder characterized by bilateral ocular oscillatory movements. Recently, mutations in FRMD7 were found to be responsible for X-linked idiopathic infantile nystagmus . We investigated the role of the FRMD7 gene mutations in seven Chinese families with infantile nystagmus. METHODS: Linkage analysis was performed with fluorescently labeled microsatellite markers, DXS1001 and DXS1047. Analysis of FRMD7 gene mutations was performed by direct sequence to the whole coding regions and exon-intron boundaries of FRMD7 gene in all affected members in seven families with IN. RESULTS: Five novel FRMD7 gene mutations, 70 G>T(p.G24W) in exon 2, c.689 690delAG (p.Ser232del) in exon8, c. 782G>A (p.R260Q) and c. 812G>T (p. C271F) in exon 9, and c. 910C>T (R303X) in exon 10, were identified in five of seven Chinese families with X-linked infantile nystagmus. But we didn't detect the FRMD7 gene mutation in one of seven families, although a positive LOD score of 2.42 (thetamax=0.1) was obtained at DXS1047 . We also found the same mutation, which is c. 782G>A (p.R260Q), occurred in two different families. CONCLUSIONS: This is first report that five kinds of FRMD7 gene mutation types occurred in Chinese families with IN, which further support that FRMD7 gene mutations are the underlying pathogenesis of the molecular mechanism for infantile nystagmus. PMID- 18431454 TI - Telemetric monitoring of 24 h intraocular pressure in conscious and freely moving C57BL/6J and CBA/CaJ mice. AB - PURPOSE: To study change patterns of 24 h intraocular pressure (IOP) in conscious and freely moving mice using telemetry. METHODS: Adult C57BL/6J and CBA/CaJ mice were entrained to a standard 12 h light and 12 h dark cycle. A telemetric pressure transmitter was implanted subcutaneously on the upper back of each light dark entrained mouse, and the pressure catheter tip was inserted into the vitreous chamber. Broadcasted IOP data were received at 120 Hz. Means of 2 min IOP were recorded every 5 min for 4-13 days to generate the 24 h IOP pattern in each mouse strain. The pattern of IOP in the C57BL/6J strain was also determined under an acute constant dark condition for 24 h. RESULTS: There were distinct patterns of 24 h IOP in the C57BL/6J and CBA/CaJ mouse strains. Under the standard light-dark condition, IOP was higher during the dark period than the light period in both strains. Elevation in IOP from the light period to the dark period was significantly smaller in the CBA/CaJ strain (1.6+/-1.7 mmHg, mean+/- standard deviation (SD), n=21) than in the C57BL/6J strain (3.4+/-2.5 mmHg, n=20). The 24 h IOP pattern in the C57BL/6J strain persisted under an acute constant dark condition (n=8). CONCLUSIONS: Distinct change patterns of 24 h IOP appeared in these two mouse strains. Although mean IOP during the dark period was significantly higher than the light period in both strains, the magnitudes of light-dark IOP elevation differed. The 24 h IOP change pattern can be driven endogenously in the absence of light. PMID- 18431455 TI - Disease mechanism for retinitis pigmentosa (RP11) caused by missense mutations in the splicing factor gene PRPF31. AB - PURPOSE: Missense mutations in the splicing factor gene PRPF31 cause a dominant form of retinitis pigmentosa (RP11) with reduced penetrance. Missense mutations in PRPF31 have previously been shown to cause reduced protein solubility, suggesting insufficiency of functional protein as the disease mechanism. Here we examine in further detail the effect of the A216P mutation on splicing function. METHODS: Splicing activity was assayed using an in vivo assay in transfected mammalian cells with rhodopsin (RHO) and transducin (GNAT1) splicing templates. Pull-down assays were used to study the interaction between PRPF31 and one of its cognate partners in the spliceosome, PRPF6. RESULTS: Splicing of RHO intron 3 and GNAT1 introns 3-5 mini-gene templates was inefficient with both spliced and unspliced products clearly detected. Assays using the RHO minigene template revealed a direct negative effect on splicing efficiency of the mutant. However, no effect of the mutation on splicing efficiency could be detected using the longer GNAT1 minigene template or using a full-length RHO transcript, splicing of which had an efficiency of 100%. No unspliced RHO transcripts could be detected in RNA from human retina. Pull-down assays between PRPF31 and PRPF6 proteins showed a stronger interaction for the mutant than wild type, suggesting a mechanism for the negative effect. CONCLUSIONS: Splicing of full-length RHO is more efficient than splicing of the minigene, and assays using a full-length template more accurately mimic splicing in photoreceptors. The RP11 missense mutations exert their pathology mainly via a mechanism based on protein insufficiency due to protein insolubility, but there is also a minor direct negative effect on function. PMID- 18431456 TI - A novel locus for X-linked congenital cataract on Xq24. AB - PURPOSE: This study aimed to map the genetic locus responsible for a novel X linked congenital cataract phenotype. METHODS: A large three-generation family with lamellar and nuclear cataract in five affected males was identified. Linkage analysis was conducted by genotyping X-chromosome specific microsatellite markers at an average spacing of 5 cM. Analysis was conducted using the LINKAGE package under an X-linked recessive model. RESULTS: A linkage was detected on Xq24 with the maximum LOD score of 2.53 at theta=0 for DXS1001. The minimal region was defined as 11.5 Mb between markers DXS8055 and DXS8009 through critical recombination events in multiple individuals. CONCLUSIONS: A gene causing this novel congenital cataract phenotype is located on the long arm of the X chromosome. PMID- 18431458 TI - Overweight and Hyperinsulinemia Provide Individual Contributions to Compromises in Brachial Artery Distensibility in Healthy Adolescents and Young Adults: Brachial Distensibility in Children. AB - BACKGROUND: Brachial artery distensibility (BrachD) was measured in healthy children to identify associations with atherosclerotic risk factors. METHODS: 969 black and white subjects 13-22 years were classified as lean (L) or overweight (O) and hyperinsulinemic (H-I) or normoinsulinemic (N-I). BP and BrachD were obtained with a DynaPulse Pathway instrument. ANOVA was performed looking for group mean differences. Correlations between BrachD and risk variables were examined. Determinates of BrachD were determined by backward elimination regression stratified by BMI-Insulin group. RESULTS: Decreased BrachD correlated with male gender, O, higher BP, HR, fasting glucose and log of fasting insulin after adjusting for pulse pressure (PP). BrachD was greatest in L/N-I with progressive decreases seen in L/H-I, O/N-I, and O/H-I subjects. Regression modeling found PP and HR were major determinates of BrachD. Glucose was significant for subjects with N-I regardless of adiposity. Excluding BP, glucose remained important in N-I subjects. Gender was significant for all. HR retained significance only in O subjects regardless of insulin level. CONCLUSIONS: In healthy adolescents, hyperinsulinemia and obesity adversely affect brachial artery function with overweight contributing to a greater degree. In normoinsulinemic subjects, fasting glucose was inversely related to BrachD. Metabolic factors may play a role in vascular function in youth. PMID- 18431459 TI - Controlling the Dimensions of Carbon Nanofiber Structures through the Electropolymerization of Pyrrole. AB - Electrically conductive polymers, such as polypyrrole (PPy), show promise for modifying the dimensions and properties of micro- and nanoscale structures. Mechanisms for controlling the formation of PPy films of nanoscale thickness were evaluated by electrochemically synthesizing and examining PPy films on planar gold electrodes under a variety of growth conditions. Tunable PPy coatings were then deposited by electropolymerization on the sidewalls of individual, electrically addressable carbon nanofibers (CNFs). The ability to modify the physical size of specific nanofibers in controllable fashion is demonstrated. The biocompatibility, potential for chemical functionalization, and ability to effect volume changes of this nanocomposite can lead to advanced functionality, such as specific, nanoscale valving of materials and morphological control at the nanoscale. PMID- 18431460 TI - Hypospadias and anorectal malformations mediated by Eph/ephrin signaling. AB - PURPOSE: Despite extensive research, the molecular basis of hypospadias and anorectal malformations is poorly understood, likely due to a multifactorial basis. The incidence of hypospadias is increasing, thus making research in this area warranted and timely. This review presents recent molecular work broadening our understanding of these disorders. MATERIALS AND METHODS: A brief review of our recent work and the literature on the role of Eph/ephrin signaling in hypospadias and anorectal malformations is presented. RESULTS: Genetically engineered mice mutant for ephrin-B2 or EphB2;EphB3 manifest a variety of genitourinary and anorectal malformations. Approximately 40% of adult male heterozygous mice demonstrate perineal hypospadias. Although homozygous mice die soon after birth, 100% of homozygous males demonstrate high imperforate anus with urethral anomalies and 100% of homozygous females demonstrate persistent cloaca. Male mice compound homozygous for EphB2(ki/ki);EphB3(Delta/Delta)/ also demonstrate hypospadias. CONCLUSIONS: These mouse models provide compelling evidence of the role of B-class Eph/ephrin signaling in genitourinary/anorectal development and add to our mechanistic and molecular understanding of normal and abnormal embryonic development. As research on the B-class Ephs and ephrins continues, they will likely be shown to be molecular contributors to the multifactorial basis of hypospadias and anorectal malformations in humans as well. PMID- 18431461 TI - HMGB1: endogenous danger signaling. AB - While foreign pathogens and their products have long been known to activate the innate immune system, the recent recognition of a group of endogenous molecules that serve a similar function has provided a framework for understanding the overlap between the inflammatory responses activated by pathogens and injury. These endogenous molecules, termed alarmins, are normal cell constituents that can be released into the extracellular milieu during states of cellular stress or damage and subsequently activate the immune system. One nuclear protein, High mobility group box-1 (HMGB1), has received particular attention as fulfilling the functions of an alarmin by being involved in both infectious and non-infectious inflammatory conditions. Once released, HMGB1 signals through various receptors to activate immune cells involved in the immune process. Although initial studies demonstrated HMGB1 as a late mediator of sepsis, recent findings indicate HMGB1 to have an important role in models of non-infectious inflammation, such as autoimmunity, cancer, trauma, and ischemia reperfusion injury. Furthermore, in contrast to its pro-inflammatory functions, there is evidence that HMGB1 also has restorative effects leading to tissue repair and regeneration. The complex functions of HMGB1 as an archetypical alarmin are outlined here to review our current understanding of a molecule that holds the potential for treatment in many important human conditions. PMID- 18431463 TI - Epidemic inflammation: pondering obesity. AB - Over the past two decades, inflammation has been recognized as a major driver in the pathogenesis of several common diseases, including atherosclerosis, diabetes, cancer, and asthma. Over the same period, there has been a steep rise in the incidence of obesity, a major risk factor for these disorders. Inflammation of adipose tissue is now recognized to accompany obesity and contribute to its sequelae. Thus, whereas obesity is primarily a disorder of energy balance, it may be helpful to consider it also as a form of epidemic inflammation that predisposes to other forms of epidemic inflammation. It is a fundamental biologic challenge to understand how a positive energy balance and inflammation are linked. This work reviews evidence that reactive oxygen and nitrogen intermediates (ROI and RNI) help drive chronic inflammation in the obese. This is proposed to be a maladaptive instance of our evolved dependence on ROI and RNI for both homeostatic signaling and host defense. ROI and RNI are well suited for these seemingly contradictory dual functions by their metabolic origin, high diffusibility in water and lipid, atomic specificity, and large number of molecular targets. When we eat so much and work so little that we repeatedly generate reactive compounds at levels normally reserved for emergencies, we treat our own cells like invading microbes. PMID- 18431462 TI - Estrogen, NFkappaB, and the heat shock response. AB - Estrogen has pleiotropic actions, among which are its anti-apoptotic, anti inflammatory, and vasodilatory effects. Recently, an interaction between 17beta estradiol (E2) and the transcription factor nuclear factor kappaB (NFkappaB) has been identified. NFkappaB has a central role in the control of genes involved in inflammation, proliferation, and apoptosis. Prolonged activation of NFkappaB is associated with numerous inflammatory pathological conditions. An important facet of E2 is its ability to modulate activity of NFkappaB via both genomic and nongenomic actions. E2 can activate NFkappaB rapidly via nongenomic pathways, increase cellular resistance to injury, and induce expression of the protective class of proteins, heat shock proteins (HSPs). HSPs can bind to many of the pro apoptotic and pro-inflammatory targets of NFkappaB and, thus, indirectly inhibit many of its deleterious effects. In addition, HSPs can block NFkappaB activation and binding directly. Similarly, genomic E2 signaling can inhibit NFkappaB, but does so through alternative mechanisms. This review focuses on the molecular mechanisms of cross-talk between E2, NFkappaB, and HSPs, and the biological relevance of this cross-talk. PMID- 18431464 TI - Lysophosphatidic acid inhibits bacterial endotoxin-induced pro-inflammatory response: potential anti-inflammatory signaling pathways. AB - Previous studies have demonstrated that heterotrimeric guanine nucleotide-binding regulatory (Gi) protein-deficient mice exhibit augmented inflammatory responses to lipopolysaccharide (LPS). These findings suggest that Gi protein agonists will suppress LPS-induced inflammatory gene expression. Lysophosphatidic acid (LPA) activates G protein-coupled receptors leading to Gi protein activation. We hypothesized that LPA will inhibit LPS-induced inflammatory responses through activation of Gi-coupled anti-inflammatory signaling pathways. We examined the anti-inflammatory effect of LPA on LPS responses both in vivo and in vitro in CD 1 mice. The mice were injected intravenously with LPA (10 mg/kg) followed by intraperitoneal injection of LPS (75 mg/kg for survival and 25 mg/kg for other studies). LPA significantly increased the mice survival to endotoxemia (P < 0.05). LPA injection reduced LPS-induced plasma TNF-alpha production (69 +/- 6%, P < 0.05) and myeloperoxidase (MPO) activity in lung (33 +/- 9%, P < 0.05) as compared to vehicle injection. LPS-induced plasma IL-6 was unchanged by LPA. In vitro studies with peritoneal macrophages paralleled results from in vivo studies. LPA (1 and 10 microM) significantly inhibited LPS-induced TNFalpha production (61 +/- 9% and 72 +/- 9%, respectively, P < 0.05) but not IL-6. We further demonstrated that the anti-inflammatory effect of LPA was reversed by ERK 1/2 and phosphatase inhibitors, suggesting that ERK 1/2 pathway and serine/threonine phosphatases are involved. Inhibition of phosphatidylinositol 3 (PI3) kinase signaling pathways also partially reversed the LPA anti-inflammatory response. However, LPA did not alter NFkappaB and peroxisome proliferator activated receptor gamma (PPARgamma) activation. Inhibitors of PPARgamma did not alter LPA-induced inhibition of LPS signaling. These studies demonstrate that LPA has significant anti-inflammatory activities involving activation of ERK 1/2, serine/threonine phosphatases, and PI3 kinase signaling pathways. PMID- 18431466 TI - SAM domain-based protein oligomerization observed by live-cell fluorescence fluctuation spectroscopy. AB - Sterile-alpha-motif (SAM) domains are common protein interaction motifs observed in organisms as diverse as yeast and human. They play a role in protein homo- and hetero-interactions in processes ranging from signal transduction to RNA binding. In addition, mutations in SAM domain and SAM-mediated oligomers have been linked to several diseases. To date, the observation of heterogeneous SAM-mediated oligomers in vivo has been elusive, which represents a common challenge in dissecting cellular biochemistry in live-cell systems. In this study, we report the oligomerization and binding stoichiometry of high-order, multi-component complexes of (SAM) domain proteins Ste11 and Ste50 in live yeast cells using fluorescence fluctuation methods. Fluorescence cross-correlation spectroscopy (FCCS) and 1-dimensional photon counting histogram (1dPCH) confirm the SAM mediated interaction and oligomerization of Ste11 and Ste50. Two-dimensional PCH (2dPCH), with endogenously expressed proteins tagged with GFP or mCherry, uniquely indicates that Ste11 and Ste50 form a heterogeneous complex in the yeast cytosol comprised of a dimer of Ste11 and a monomer of Ste50. In addition, Ste50 also exists as a high order oligomer that does not interact with Ste11, and the size of this oligomer decreases in response to signals that activate the MAP kinase cascade. Surprisingly, a SAM domain mutant of Ste50 disrupted not only the Ste50 oligomers but also Ste11 dimerization. These results establish an in vivo model of Ste50 and Ste11 homo- and hetero-oligomerization and highlight the usefulness of 2dPCH for quantitative dissection of complex molecular interactions in genetic model organisms such as yeast. PMID- 18431467 TI - Genome Reshuffling for Advanced Intercross Permutation (GRAIP): simulation and permutation for advanced intercross population analysis. AB - BACKGROUND: Advanced intercross lines (AIL) are segregating populations created using a multi-generation breeding protocol for fine mapping complex trait loci (QTL) in mice and other organisms. Applying QTL mapping methods for intercross and backcross populations, often followed by naive permutation of individuals and phenotypes, does not account for the effect of AIL family structure in which final generations have been expanded and leads to inappropriately low significance thresholds. The critical problem with naive mapping approaches in AIL populations is that the individual is not an exchangeable unit. METHODOLOGY/PRINCIPAL FINDINGS: The effect of family structure has immediate implications for the optimal AIL creation (many crosses, few animals per cross, and population expansion before the final generation) and we discuss these and the utility of AIL populations for QTL fine mapping. We also describe Genome Reshuffling for Advanced Intercross Permutation, (GRAIP) a method for analyzing AIL data that accounts for family structure. GRAIP permutes a more interchangeable unit in the final generation crosses - the parental genome - and simulating regeneration of a permuted AIL population based on exchanged parental identities. GRAIP determines appropriate genome-wide significance thresholds and locus-specific P-values for AILs and other populations with similar family structures. We contrast GRAIP with naive permutation using a large densely genotyped mouse AIL population (1333 individuals from 32 crosses). A naive permutation using coat color as a model phenotype demonstrates high false positive locus identification and uncertain significance levels, which are corrected using GRAIP. GRAIP also detects an established hippocampus weight locus and a new locus, Hipp9a. CONCLUSIONS AND SIGNIFICANCE: GRAIP determines appropriate genome-wide significance thresholds and locus-specific P-values for AILs and other populations with similar family structures. The effect of family structure has immediate implications for the optimal AIL creation and we discuss these and the utility of AIL populations. PMID- 18431468 TI - ESAT-6/CFP10 skin test predicts disease in M. tuberculosis-infected guinea pigs. AB - BACKGROUND: Targeted preventive chemotherapy of individuals with progressive subclinical (incipient) disease before it becomes contagious would break the chain of tuberculosis transmission in high endemic regions. We have studied the ability of a skin test response to ESAT-6 and CFP10 (E6/C10) to predict later development of tuberculosis disease in the guinea pig model. METHODS AND FINDINGS: Guinea pigs, either vaccinated with BCG or unvaccinated, were infected with a low dose of Mycobacterium tuberculosis by the aerosol route and the development of delayed type hypersensitivity responses to E6/C10 and to purified protein derivative (PPD) were followed until the onset of clinical disease. We demonstrated a negative correlation between the size of the skin test response and the time to the onset of clinical disease; a large E6/C10 skin test response correlated to a shorter survival time post skin testing, while a small E6/C10 skin test reaction correlated with a longer survival time (r = -0.6 and P<0.0001). No correlation was found using PPD. CONCLUSIONS: Our data suggest that it may be possible to develop a prognostic skin test based on E6/C10 that will allow the identification of individuals with incipient disease, who have the highest risk of developing active tuberculosis in the near future. PMID- 18431469 TI - Marine incursion: the freshwater herring of Lake Tanganyika are the product of a marine invasion into west Africa. AB - The spectacular marine-like diversity of the endemic fauna of Lake Tanganyika, the oldest of the African Great Lakes, led early researchers to suggest that the lake must have once been connected to the ocean. Recent geophysical reconstructions clearly indicate that Lake Tanganyika formed by rifting in the African subcontinent and was never directly linked to the sea. Although the Lake has a high proportion of specialized endemics, the absence of close relatives outside Tanganyika has complicated phylogeographic reconstructions of the timing of lake colonization and intralacustrine diversification. The freshwater herring of Lake Tanganyika are members of a large group of pellonuline herring found in western and southern Africa, offering one of the best opportunities to trace the evolutionary history of members of Tanganyika's biota. Molecular phylogenetic reconstructions indicate that herring colonized West Africa 25-50MYA, at the end of a major marine incursion in the region. Pellonuline herring subsequently experienced an evolutionary radiation in West Africa, spreading across the continent and reaching East Africa's Lake Tanganyika during its early formation. While Lake Tanganyika has never been directly connected with the sea, the endemic freshwater herring of the lake are the descendents of an ancient marine incursion, a scenario which may also explain the origin of other Tanganyikan endemics. PMID- 18431470 TI - The T7-primer is a source of experimental bias and introduces variability between microarray platforms. AB - Eberwine(-like) amplification of mRNA adds distinct 6-10 bp nucleotide stretches to the 5' end of amplified RNA transcripts. Analysis of over six thousand microarrays reveals that probes containing motifs complementary to these stretches are associated with aberrantly high signals up to a hundred fold the signal observed in unaffected probes. This is not observed when total RNA is used as target source. Different T7 primer sequences are used in different laboratories and platforms and consequently different T7 primer bias is observed in different datasets. This will hamper efforts to compare data sets across platforms. PMID- 18431471 TI - Antihypertensive drug guanabenz is active in vivo against both yeast and mammalian prions. AB - BACKGROUND: Prion-based diseases are incurable transmissible neurodegenerative disorders affecting animals and humans. METHODOLOGY/PRINCIPAL FINDINGS: Here we report the discovery of the in vivo antiprion activity of Guanabenz (GA), an agonist of alpha2-adrenergic receptors routinely used in human medicine as an antihypertensive drug. We isolated GA in a screen for drugs active in vivo against two different yeast prions using a previously described yeast-based two steps assay. GA was then shown to promote ovine PrP(Sc) clearance in a cell-based assay. These effects are very specific as evidenced by the lack of activity of some GA analogues that we generated. GA antiprion activity does not involve its agonist activity on alpha2-adrenergic receptors as other chemically close anti hypertensive agents possessing related mechanism of action were found inactive against prions. Finally, GA showed activity in a transgenic mouse-based in vivo assay for ovine prion propagation, prolonging slightly but significantly the survival of treated animals. CONCLUSION/SIGNIFICANCE: GA thus adds to the short list of compounds active in vivo in animal models for the treatment of prion based diseases. Because it has been administrated for many years to treat hypertension on a daily basis, without major side-effects, our results suggest that it could be evaluated in human as a potential treatment for prion-based diseases. PMID- 18431472 TI - Default Pathway of var2csa switching and translational repression in Plasmodium falciparum. AB - Antigenic variation is a subtle process of fundamental importance to the survival of a microbial pathogen. In Plasmodium falciparum malaria, PfEMP1 is the major variable antigen and adhesin expressed at the surface of the infected erythrocyte, which is encoded for by members of a family of 60 var-genes. Peri nuclear repositioning and epigenetic mechanisms control their mono-allelic expression. The switching of PfEMP1 depends in part on variable transition rates and short-lived immune responses to shared minor epitopes. Here we show var-genes to switch to a common gene that is highly transcribed, but sparsely translated into PfEMP1 and not expressed at the erythrocyte surface. Highly clonal and adhesive P. falciparum, which expressed distinct var-genes and the corresponding PfEMP1s at onset, were propagated without enrichment or panning. The parasites successively and spontaneously switched to transcribe a shared var-gene (var2csa) matched by the loss of PfEMP1 surface expression and host cell-binding. The var2csa gene repositioned in the peri-nuclear area upon activation, away from the telomeric clusters and heterochromatin to transcribe spliced, full-length RNA. Despite abundant transcripts, the level of intracellular PfEMP1 was low suggesting post-transcriptional mechanisms to partake in protein expression. In vivo, off-switching and translational repression may constitute one pathway, among others, coordinating PfEMP1 expression. PMID- 18431473 TI - Treg depletion inhibits efficacy of cancer immunotherapy: implications for clinical trials. AB - BACKGROUND: Regulatory T lymphocytes (Treg) infiltrate human glioblastoma (GBM); are involved in tumor progression and correlate with tumor grade. Transient elimination of Tregs using CD25 depleting antibodies (PC61) has been found to mediate GBM regression in preclinical models of brain tumors. Clinical trials that combine Treg depletion with tumor vaccination are underway to determine whether transient Treg depletion can enhance anti-tumor immune responses and improve long term survival in cancer patients. FINDINGS: Using a syngeneic intracrabial glioblastoma (GBM) mouse model we show that systemic depletion of Tregs 15 days after tumor implantation using PC61 resulted in a decrease in Tregs present in tumors, draining lymph nodes and spleen and improved long-term survival (50% of mice survived >150 days). No improvement in survival was observed when Tregs were depleted 24 days after tumor implantation, suggesting that tumor burden is an important factor for determining efficacy of Treg depletion in clinical trials. In a T cell dependent model of brain tumor regression elicited by intratumoral delivery of adenoviral vectors (Ad) expressing Fms-like Tyrosine Kinase 3 ligand (Flt3L) and Herpes Simplex Type 1 Thymidine Kinase (TK) with ganciclovir (GCV), we demonstrate that administration of PC61 24 days after tumor implantation (7 days after treatment) inhibited T cell dependent tumor regression and long term survival. Further, depletion with PC61 completely inhibited clonal expansion of tumor antigen-specific T lymphocytes in response to the treatment. CONCLUSIONS: Our data demonstrate for the first time, that although Treg depletion inhibits the progression/eliminates GBM tumors, its efficacy is dependent on tumor burden. We conclude that this approach will be useful in a setting of minimal residual disease. Further, we also demonstrate that Treg depletion, using PC61 in combination with immunotherapy, inhibits clonal expansion of tumor antigen-specific T cells, suggesting that new, more specific targets to block Tregs will be necessary when used in combination with therapies that activate anti-tumor immunity. PMID- 18431474 TI - Serum antibodies to Porphyromonas gingivalis chaperone HtpG predict health in periodontitis susceptible patients. AB - BACKGROUND: Chaperones are ubiquitous conserved proteins critical in stabilization of new proteins, repair/removal of defective proteins and immunodominant antigens in innate and adaptive immunity. Periodontal disease is a chronic inflammatory infection associated with infection by Porphyromonas gingivalis that culminates in the destruction of the supporting structures of the teeth. We previously reported studies of serum antibodies reactive with the human chaperone Hsp90 in gingivitis, a reversible form of gingival disease confined to the oral soft tissues. In those studies, antibodies were at their highest levels in subjects with the best oral health. We hypothesized that antibodies to the HSP90 homologue of P. gingivalis (HtpG) might be associated with protection/resistance against destructive periodontitis. METHODOLOGY/PRINCIPAL FINDINGS: ELISA assays using cloned HtpG and peptide antigens confirmed gingivitis subjects colonized with P. gingivalis had higher serum levels of anti HtpG and, concomitantly, lower levels of attachment loss. Additionally, serum antibody levels to P. gingivalis HtpG protein were higher in healthy subjects compared to patients with either chronic or aggressive periodontitis. We found a negative association between tooth attachment loss and anti-P. gingivalis HtpG (p = 0.043) but not anti-Fusobacterium nucleatum (an oral opportunistic commensal) HtpG levels. Furthermore, response to periodontal therapy was more successful in subjects having higher levels of anti-P. gingivalis HtpG before treatment (p = 0.018). There was no similar relationship to anti-F. nucleatum HtpG levels. Similar results were obtained when these experiments were repeated with a synthetic peptide of a region of P. gingivalis HtpG. CONCLUSIONS/SIGNIFICANCE: OUR RESULTS SUGGEST: 1) anti-P. gingivalis HtpG antibodies are protective and therefore predict health periodontitis-susceptable patients; 2) may augment the host defence to periodontitis and 3) a unique peptide of P. gingivalis HtpG offers significant potential as an effective diagnostic target and vaccine candidate. These results are compatible with a novel immune control mechanism unrelated to direct binding of bacteria. PMID- 18431475 TI - Interactive "Video Doctor" counseling reduces drug and sexual risk behaviors among HIV-positive patients in diverse outpatient settings. AB - BACKGROUND: Reducing substance use and unprotected sex by HIV-positive persons improves individual health status while decreasing the risk of HIV transmission. Despite recommendations that health care providers screen and counsel their HIV positive patients for ongoing behavioral risks, it is unknown how to best provide "prevention with positives" in clinical settings. Positive Choice, an interactive, patient-tailored computer program, was developed in the United States to improve clinic-based assessment and counseling for risky behaviors. METHODOLOGY AND FINDINGS: We conducted a parallel groups randomized controlled trial (December 2003-September 2006) at 5 San Francisco area outpatient HIV clinics. Eligible patients (HIV-positive English-speaking adults) completed an in depth computerized risk assessment. Participants reporting substance use or sexual risks (n = 476) were randomized in stratified blocks. The intervention group received tailored risk-reduction counseling from a "Video Doctor" via laptop computer and a printed Educational Worksheet; providers received a Cueing Sheet on reported risks. Compared with control, fewer intervention participants reported continuing illicit drug use (RR 0.81, 95% CI: 0.689, 0.957, p = 0.014 at 3 months; and RR 0.65, 95% CI: 0.540, 0.785, p<0.001 at 6 months) and unprotected sex (RR 0.88, 95% CI: 0.773, 0.993, p = 0.039 at 3 months; and RR 0.80, 95% CI: 0.686, 0.941, p = 0.007 at 6 months). Intervention participants reported fewer mean days of ongoing illicit drug use (-4.0 days vs. -1.3 days, p = 0.346, at 3 months; and -4.7 days vs. -0.7 days, p = 0.130, at 6 months) than did controls, and had fewer casual sex partners at (-2.3 vs. -1.4, p = 0.461, at 3 months; and 2.7 vs. -0.6, p = 0.042, at 6 months). CONCLUSIONS: The Positive Choice intervention achieved significant cessation of illicit drug use and unprotected sex at the group-level, and modest individual-level reductions in days of ongoing drug use and number of casual sex partners compared with the control group. Positive Choice, including Video Doctor counseling, is an efficacious and appropriate adjunct to risk-reduction efforts in outpatient settings, and holds promise as a public health HIV intervention. TRIAL REGISTRATION: Clinicaltrials.gov NCT00447707. PMID- 18431476 TI - Inferring microRNA activities by combining gene expression with microRNA target prediction. AB - BACKGROUND: MicroRNAs (miRNAs) play crucial roles in a variety of biological processes via regulating expression of their target genes at the mRNA level. A number of computational approaches regarding miRNAs have been proposed, but most of them focus on miRNA gene finding or target predictions. Little computational work has been done to investigate the effective regulation of miRNAs. METHODOLOGY/PRINCIPAL FINDINGS: We propose a method to infer the effective regulatory activities of miRNAs by integrating microarray expression data with miRNA target predictions. The method is based on the idea that regulatory activity changes of miRNAs could be reflected by the expression changes of their target transcripts measured by microarray. To validate this method, we apply it to the microarray data sets that measure gene expression changes in cell lines after transfection or inhibition of several specific miRNAs. The results indicate that our method can detect activity enhancement of the transfected miRNAs as well as activity reduction of the inhibited miRNAs with high sensitivity and specificity. Furthermore, we show that our inference is robust with respect to false positives of target prediction. CONCLUSIONS/SIGNIFICANCE: A huge amount of gene expression data sets are available in the literature, but miRNA regulation underlying these data sets is largely unknown. The method is easy to be implemented and can be used to investigate the miRNA effective regulation underlying the expression change profiles obtained from microarray experiments. PMID- 18431477 TI - Distinct haptic cues do not reduce interference when learning to reach in multiple force fields. AB - BACKGROUND: Previous studies of learning to adapt reaching movements in the presence of novel forces show that learning multiple force fields is prone to interference. Recently it has been suggested that force field learning may reflect learning to manipulate a novel object. Within this theoretical framework, interference in force field learning may be the result of static tactile or haptic cues associated with grasp, which fail to indicate changing dynamic conditions. The idea that different haptic cues (e.g. those associated with different grasped objects) signal motor requirements and promote the learning and retention of multiple motor skills has previously been unexplored in the context of force field learning. METHODOLOGY/PRINCIPLE FINDINGS: The present study tested the possibility that interference can be reduced when two different force fields are associated with differently shaped objects grasped in the hand. Human subjects were instructed to guide a cursor to targets while grasping a robotic manipulandum, which applied two opposing velocity-dependent curl fields to the hand. For one group of subjects the manipulandum was fitted with two different handles, one for each force field. No attenuation in interference was observed in these subjects relative to controls who used the same handle for both force fields. CONCLUSIONS/SIGNIFICANCE: These results suggest that in the context of the present learning paradigm, haptic cues on their own are not sufficient to reduce interference and promote learning multiple force fields. PMID- 18431478 TI - Does 3-day course of oral amoxycillin benefit children of non-severe pneumonia with wheeze: a multicentric randomised controlled trial. AB - BACKGROUND: WHO-defined pneumonias, treated with antibiotics, are responsible for a significant proportion of childhood morbidity and mortality in the developing countries. Since substantial proportion pneumonias have a viral etiology, where children are more likely to present with wheeze, there is a concern that currently antibiotics are being over-prescribed for it. Hence the current trial was conducted with the objective to show the therapeutic equivalence of two treatments (placebo and amoxycillin) for children presenting with non-severe pneumonia with wheeze, who have persistent fast breathing after nebulisation with salbutamol, and have normal chest radiograph. METHODOLOGY: This multi-centric, randomised placebo controlled double blind clinical trial intended to investigate equivalent efficacy of placebo and amoxicillin and was conducted in ambulatory care settings in eight government hospitals in India. Participants were children aged 2-59 months of age, who received either oral amoxycillin (31-54 mg/Kg/day, in three divided doses for three days) or placebo, and standard bronchodilator therapy. Primary outcome was clinical failure on or before day- 4. PRINCIPAL FINDINGS: We randomized 836 cases in placebo and 835 in amoxycillin group. Clinical failures occurred in 201 (24.0%) on placebo and 166 (19.9%) on amoxycillin (risk difference 4.2% in favour of antibiotic, 95% CI: 0.2 to 8.1). Adherence for both placebo and amoxycillin was >96% and 98.9% subjects were followed up on day- 4. Clinical failure was associated with (i) placebo treatment (adjusted OR = 1.28, 95% CI: 1.01 to1.62), (ii) excess respiratory rate of >10 breaths per minute (adjusted OR = 1.51, 95% CI: 1.19, 1.92), (iii) vomiting at enrolment (adjusted OR = 1.49, 95% CI: 1.13, 1.96), (iv) history of use of broncho-dilators (adjusted OR = 1.71, 95% CI: 1.30, 2.24) and (v) non-adherence (adjusted OR = 8.06, 95% CI: 4.36, 14.92). CONCLUSIONS: Treating children with non-severe pneumonia and wheeze with a placebo is not equivalent to treatment with oral amoxycillin. TRIAL REGISTRATION: ClinicalTrials.gov NCT00407394. PMID- 18431479 TI - Truncated power laws reveal a link between low-level behavioral processes and grouping patterns in a colonial bird. AB - BACKGROUND: Departures from power law group size frequency distributions have been proposed as a useful tool to link individual behavior with population patterns and dynamics, although examples are scarce for wild animal populations. METHODOLOGY/PRINCIPAL FINDINGS: We studied a population of Lesser kestrels (Falco naumanni) breeding in groups (colonies) from one to ca. 40 breeding pairs in 10,000 km(2) in NE Spain. A 3.5 fold steady population increase occurred during the eight-year study period, accompanied by a geographical expansion from an initial subpopulation which in turn remained stable in numbers. This population instability was mainly driven by first-breeders, which are less competitive at breeding sites, being relegated to breed solitarily or in small colony sizes, and disperse farther than adults. Colony size frequency distributions shifted from an initial power law to a truncated power law mirroring population increase. Thus, we hypothesized that population instability was behind the truncation of the power law. Accordingly, we found a power law distribution through years in the initial subpopulation, and a match between the power law breakpoint (at ca. ten pairs) and those colony sizes from which the despotic behavior of colony owners started to impair the settlement of newcomers. Moreover, the instability hypothesis was further supported by snapshot data from another population of Lesser kestrels in SW Spain suffering a population decline. CONCLUSIONS/SIGNIFICANCE: Appropriate analysis of the scaling properties of grouping patterns has unraveled the link between local agonistic processes and large-scale (population) grouping patterns in a wild bird population. PMID- 18431480 TI - Intradermally administered yellow fever vaccine at reduced dose induces a protective immune response: a randomized controlled non-inferiority trial. AB - BACKGROUND: Implementation of yellow fever vaccination is currently hampered by limited supply of vaccine. An alternative route of administration with reduced amounts of vaccine but without loss of vaccine efficacy would boost vaccination programmes. METHODS AND FINDINGS: A randomized, controlled, non-inferiority trial was conducted in a Dutch university center between August 2005 and February 2007. A total of 155 primary vaccinated and 20 previously vaccinated volunteers participated. Participants were randomly assigned in a 1ratio1 ratio to receive intradermal (i.d.) vaccination with live attenuated yellow fever 17D vaccine at a reduced dose (1/5(th); 0.1 mL) or the conventional subcutaneous (s.c.) vaccination (0.5 mL). Antibody neutralization titers were determined at 2, 4 and 8 weeks and 1 year after vaccination by counting the reduction in virus-induced plaques in the presence of serial serum dilutions. Adverse events were documented in a 3-week dairy. Viraemia was measured 5 days after vaccination. From 2 weeks up to one year after vaccination, the maximum serum-dilution at which 80% of the virus plaques were neutralized, which indicates protection against yellow fever, did not differ between those given a reduced i.d. dose or standard s.c. dose of vaccine. In all cases the WHO standard of seroprotection (i.e. 80% virus neutralization) was reached (in 77/77 and 78/78, respectively). Similar results were found in the previously vaccinated individuals. Viraemia was detected in half of the primary vaccinated participants, which was not predictive of serological response. In revaccinees no viraemia was detected. CONCLUSIONS: Intradermal administration of one fifth of the amount of yellow fever vaccine administered subcutaneously results in protective seroimmunity in all volunteers. Albeit this vaccination route should enable vaccination of five-times as many individuals at risk for disease, these results should now be confirmed in field studies in areas with potential yellow fever virus transmission to change vaccination policy. TRIAL REGISTRATION: Nederlands Trial Register ISRCTN46326316. PMID- 18431481 TI - Sorting signals, N-terminal modifications and abundance of the chloroplast proteome. AB - Characterization of the chloroplast proteome is needed to understand the essential contribution of the chloroplast to plant growth and development. Here we present a large scale analysis by nanoLC-Q-TOF and nanoLC-LTQ-Orbitrap mass spectrometry (MS) of ten independent chloroplast preparations from Arabidopsis thaliana which unambiguously identified 1325 proteins. Novel proteins include various kinases and putative nucleotide binding proteins. Based on repeated and independent MS based protein identifications requiring multiple matched peptide sequences, as well as literature, 916 nuclear-encoded proteins were assigned with high confidence to the plastid, of which 86% had a predicted chloroplast transit peptide (cTP). The protein abundance of soluble stromal proteins was calculated from normalized spectral counts from LTQ-Obitrap analysis and was found to cover four orders of magnitude. Comparison to gel-based quantification demonstrates that 'spectral counting' can provide large scale protein quantification for Arabidopsis. This quantitative information was used to determine possible biases for protein targeting prediction by TargetP and also to understand the significance of protein contaminants. The abundance data for 550 stromal proteins was used to understand abundance of metabolic pathways and chloroplast processes. We highlight the abundance of 48 stromal proteins involved in post-translational proteome homeostasis (including aminopeptidases, proteases, deformylases, chaperones, protein sorting components) and discuss the biological implications. N-terminal modifications were identified for a subset of nuclear- and chloroplast encoded proteins and a novel N-terminal acetylation motif was discovered. Analysis of cTPs and their cleavage sites of Arabidopsis chloroplast proteins, as well as their predicted rice homologues, identified new species-dependent features, which will facilitate improved subcellular localization prediction. No evidence was found for suggested targeting via the secretory system. This study provides the most comprehensive chloroplast proteome analysis to date and an expanded Plant Proteome Database (PPDB) in which all MS data are projected on identified gene models. PMID- 18431482 TI - Automated three-dimensional detection and shape classification of dendritic spines from fluorescence microscopy images. AB - A fundamental challenge in understanding how dendritic spine morphology controls learning and memory has been quantifying three-dimensional (3D) spine shapes with sufficient precision to distinguish morphologic types, and sufficient throughput for robust statistical analysis. The necessity to analyze large volumetric data sets accurately, efficiently, and in true 3D has been a major bottleneck in deriving reliable relationships between altered neuronal function and changes in spine morphology. We introduce a novel system for automated detection, shape analysis and classification of dendritic spines from laser scanning microscopy (LSM) images that directly addresses these limitations. The system is more accurate, and at least an order of magnitude faster, than existing technologies. By operating fully in 3D the algorithm resolves spines that are undetectable with standard two-dimensional (2D) tools. Adaptive local thresholding, voxel clustering and Rayburst Sampling generate a profile of diameter estimates used to classify spines into morphologic types, while minimizing optical smear and quantization artifacts. The technique opens new horizons on the objective evaluation of spine changes with synaptic plasticity, normal development and aging, and with neurodegenerative disorders that impair cognitive function. PMID- 18431483 TI - Understanding water equilibration fundamentals as a step for rational protein crystallization. AB - BACKGROUND: Vapor diffusion is the most widely used technique for protein crystallization and the rate of water evaporation plays a key role on the quality of the crystals. Attempts have been made in the past to solve the mass transfer problem governing the evaporation process, either analytically or by employing numerical methods. Despite these efforts, the methods used for protein crystallization remain based on trial and error techniques rather than on fundamental principles. METHODOLOGY/PRINCIPAL FINDINGS: Here we present a new theoretical model which describes the hanging drop method as a function of the different variables that are known to influence the evaporation process. The model is extensively tested against experimental data published by other authors and considering different crystallizing conditions. Aspects responsible for the discrepancies between the existing theories and the measured evaporation kinetics are especially discussed; they include the characterization of vapor-liquid equilibrium, the role of mass transfer within the evaporating droplet, and the influence of the droplet-reservoir distance. CONCLUSIONS/SIGNIFICANCE: The validation tests show that the proposed model can be used to predict the water evaporation rates under a wide range of experimental conditions used in the hanging drop vapor-diffusion method, with no parameter fitting or computational requirements. This model combined with protein solubility data is expected to become a useful tool for a priori screening of crystallization conditions. PMID- 18431484 TI - Anti-HIV activity mediated by natural killer and CD8+ cells after toll-like receptor 7/8 triggering. AB - We previously found that triggering TLR7/8 either by single stranded HIV RNA or synthetic compounds induced changes in the lymphoid microenvironment unfavorable to HIV. In this study, we used selective TLR7 and 8 agonists to dissect their contribution to the anti-HIV effects. While triggering TLR7 inhibited efficiently HIV replication in lymphoid suspension cells from tonsillar origin, its effect was inconsistent in peripheral blood mononuclear cells (PBMC). In contrast, triggering TLR8 showed a very prominent and overall very consistent effect in PBMC and tonsillar lymphoid suspension cells. Depletion of dendritic cells (DC), Natural killer cells (NK) and CD8+ T-cells from PBMC resulted in the reversal of TLR8 induced anti-HIV effects. Especially noteworthy, depletion of either NK or CD8+ T-cells alone was only partially effective. We interpret these findings that DC are the initiator of complex changes in the microenvironment that culminates in the anti-HIV active NK and CD8+ effector cells. The near lack of NK and the low number of CD8+ T-cells in tonsillar lymphoid suspension cells may explain the lower TLR8 agonist's anti-HIV effects in that tissue. However, additional cell type specific differences must exist since the TLR7 agonists had a very strong inhibitory effect in tonsillar lymphoid suspension cells. Separation of effector from the CD4+ target cells did not abolish the anti-HIV effects pointing to the critical role of soluble factors. Triggering TLR7 or 8 were accompanied by major changes in the cytokine milieu; however, it appeared that not a single soluble factor could be assigned for the potent effects. These results delineate the complex effects of triggering TLR7/8 for an efficient antiviral defense. While the ultimate mechanism(s) remains unknown, the potent effects described may have therapeutic value for treating chronic viral diseases. Notably, HIV replication is blocked by TLR triggering before HIV integrates into the host chromosome which would prevent the establishment or maintenance of the latent reservoir. PMID- 18431485 TI - Genetic determination and linkage mapping of Plasmodium falciparum malaria related traits in Senegal. AB - Plasmodium falciparum malaria episodes may vary considerably in their severity and clinical manifestations. There is good evidence that host genetic factors contribute to this variability. To date, most genetic studies aiming at the identification of these genes have used a case/control study design for severe malaria, exploring specific candidate genes. Here, we performed a family-based genetic study of falciparum malaria related phenotypes in two independent longitudinal survey cohorts, as a first step towards the identification of genes and mechanisms involved in the outcome of infection. We studied two Senegalese villages, Dielmo and Ndiop that differ in ethnicity, malaria transmission and endemicity. We performed genome-scan linkage analysis of several malaria-related phenotypes both during clinical attacks and asymptomatic infection. We show evidence for a strong genetic contribution to both the number of clinical falciparum malaria attacks and the asymptomatic parasite density. The asymptomatic parasite density showed linkage to chromosome 5q31 (LOD = 2.26, empirical p = 0.0014, Dielmo), confirming previous findings in other studies. Suggestive linkage values were also obtained at three additional chromosome regions: the number of clinical malaria attacks on chromosome 5p15 (LOD = 2.57, empirical p = 0.001, Dielmo) and 13q13 (LOD = 2.37, empirical p = 0.0014 Dielmo), and the maximum parasite density during asymptomatic infection on chromosome 12q21 (LOD = 3.1, empirical p<10(-4), Ndiop). While regions of linkage show little overlap with genes known to be involved in severe malaria, the four regions appear to overlap with regions linked to asthma or atopy related traits, suggesting that common immune related pathways may be involved. PMID- 18431486 TI - Normalized cut group clustering of resting-state FMRI data. AB - BACKGROUND: Functional brain imaging studies have indicated that distinct anatomical brain regions can show coherent spontaneous neuronal activity during rest. Regions that show such correlated behavior are said to form resting-state networks (RSNs). RSNs have been investigated using seed-dependent functional connectivity maps and by using a number of model-free methods. However, examining RSNs across a group of subjects is still a complex task and often involves human input in selecting meaningful networks. METHODOLOGY/PRINCIPAL FINDINGS: We report on a voxel based model-free normalized cut graph clustering approach with whole brain coverage for group analysis of resting-state data, in which the number of RSNs is computed as an optimal clustering fit of the data. Inter-voxel correlations of time-series are grouped at the individual level and the consistency of the resulting networks across subjects is clustered at the group level, defining the group RSNs. We scanned a group of 26 subjects at rest with a fast BOLD sensitive fMRI scanning protocol on a 3 Tesla MR scanner. CONCLUSIONS/SIGNIFICANCE: An optimal group clustering fit revealed 7 RSNs. The 7 RSNs included motor/visual, auditory and attention networks and the frequently reported default mode network. The found RSNs showed large overlap with recently reported resting-state results and support the idea of the formation of spatially distinct RSNs during rest in the human brain. PMID- 18431487 TI - Damaged DNA binding protein 2 plays a role in breast cancer cell growth. AB - The Damaged DNA binding protein 2 (DDB2), is involved in nucleotide excision repair as well as in other biological processes in normal cells, including transcription and cell cycle regulation. Loss of DDB2 function may be related to tumor susceptibility. However, hypothesis of this study was that DDB2 could play a role in breast cancer cell growth, resulting in its well known interaction with the proliferative marker E2F1 in breast neoplasia. DDB2 gene was overexpressed in estrogen receptor (ER)-positive (MCF-7 and T47D), but not in ER-negative breast cancer (MDA-MB231 and SKBR3) or normal mammary epithelial cell lines. In addition, DDB2 expression was significantly (3.0-fold) higher in ER-positive than in ER-negative tumor samples (P = 0.0208) from 16 patients with breast carcinoma. Knockdown of DDB2 by small interfering RNA in MCF-7 cells caused a decrease in cancer cell growth and colony formation. Inversely, introduction of the DDB2 gene into MDA-MB231 cells stimulated growth and colony formation. Cell cycle distribution and 5 Bromodeoxyuridine incorporation by flow cytometry analysis showed that the growth-inhibiting effect of DDB2 knockdown was the consequence of a delayed G1/S transition and a slowed progression through the S phase of MCF-7 cells. These results were supported by a strong decrease in the expression of S phase markers (Proliferating Cell Nuclear Antigen, cyclin E and dihydrofolate reductase). These findings demonstrate for the first time that DDB2 can play a role as oncogene and may become a promising candidate as a predictive marker in breast cancer. PMID- 18431488 TI - Floral temperature and optimal foraging: is heat a feasible floral reward for pollinators? AB - As well as nutritional rewards, some plants also reward ectothermic pollinators with warmth. Bumble bees have some control over their temperature, but have been shown to forage at warmer flowers when given a choice, suggesting that there is some advantage to them of foraging at warm flowers (such as reducing the energy required to raise their body to flight temperature before leaving the flower). We describe a model that considers how a heat reward affects the foraging behaviour in a thermogenic central-place forager (such as a bumble bee). We show that although the pollinator should spend a longer time on individual flowers if they are warm, the increase in total visit time is likely to be small. The pollinator's net rate of energy gain will be increased by landing on warmer flowers. Therefore, if a plant provides a heat reward, it could reduce the amount of nectar it produces, whilst still providing its pollinator with the same net rate of gain. We suggest how heat rewards may link with plant life history strategies. PMID- 18431489 TI - Digestive duet: midgut digestive proteinases of Manduca sexta ingesting Nicotiana attenuata with manipulated trypsin proteinase inhibitor expression. AB - BACKGROUND: The defensive effect of endogenous trypsin proteinase inhibitors (NaTPIs) on the herbivore Manduca sexta was demonstrated by genetically altering NaTPI production in M. sexta's host plant, Nicotiana attenuata. To understand how this defense works, we studied the effects of NaTPI on M. sexta gut proteinase activity levels in different larval instars of caterpillars feeding freely on untransformed and transformed plants. METHODOLOGY/ PRINCIPAL FINDINGS: Second and third instars larvae that fed on NaTPI-producing (WT) genotypes were lighter and had less gut proteinase activity compared to those that fed on genotypes with either little or no NaTPI activity. Unexpectedly, NaTPI activity in vitro assays not only inhibited the trypsin sensitive fraction of gut proteinase activity but also halved the NaTPI-insensitive fraction in third-instar larvae. Unable to degrade NaTPI, larvae apparently lacked the means to adapt to NaTPI in their diet. However, caterpillars recovered at least part of their gut proteinase activity when they were transferred from NaTPI-producing host plants to NaTPI free host plants. In addition extracts of basal leaves inhibited more gut proteinase activity than did extracts of middle stem leaves with the same protein content. CONCLUSIONS/ SIGNIFICANCE: Although larvae can minimize the effects of high NaTPI levels by feeding on leaves with high protein and low NaTPI activity, the host plant's endogenous NaTPIs remain an effective defense against M. sexta, inhibiting gut proteinase and affecting larval performance. PMID- 18431491 TI - The effect of isovolemic hemodilution with oxycyte, a perfluorocarbon emulsion, on cerebral blood flow in rats. AB - BACKGROUND: Cerebral blood flow (CBF) is auto-regulated to meet the brain's metabolic requirements. Oxycyte is a perfluorocarbon emulsion that acts as a highly effective oxygen carrier compared to blood. The aim of this study is to determine the effects of Oxycyte on regional CBF (rCBF), by evaluating the effects of stepwise isovolemic hemodilution with Oxycyte on CBF. METHODOLOGY: Male rats were intubated and ventilated with 100% O(2) under isoflurane anesthesia. The regional (striatum) CBF (rCBF) was measured with a laser doppler flowmeter (LDF). Stepwise isovolemic hemodilution was performed by withdrawing 4ml of blood and substituting the same volume of 5% albumin or 2 ml Oxycyte plus 2 ml albumin at 20-minute intervals until the hematocrit (Hct) values reached 5%. PRINCIPAL FINDINGS: In the albumin-treated group, rCBF progressively increased to approximately twice its baseline level (208+/-30%) when Hct levels were less than 10%. In the Oxycyte-treated group on the other hand, rCBF increased by significantly smaller increments, and this group's mean rCBF was only slightly higher than baseline (118+/-18%) when Hct levels were less than 10%. Similarly, in the albumin-treated group, rCBF started to increase when hemodilution with albumin caused the CaO(2) to decrease below 17.5 ml/dl. Thereafter, the increase in rCBF was accompanied by a nearly proportional decrease in the CaO(2) level. In the Oxycyte-treated group, the increase in rCBF was significantly smaller than in the albumin-treated group when the CaO(2) level dropped below 10 ml/dl (142+/-20% vs. 186+/-26%), and rCBF returned to almost baseline levels (106+/-15) when the CaO(2) level was below 7 ml/dl. CONCLUSIONS/SIGNIFICANCE: Hemodilution with Oxycyte was accompanied with higher CaO(2) and PO(2) than control group treated with albumin alone. This effect may be partially responsible for maintaining relatively constant CBF and not allowing the elevated blood flow that was observed with albumin. PMID- 18431492 TI - Nutrition or detoxification: why bats visit mineral licks of the Amazonian rainforest. AB - Many animals in the tropics of Africa, Asia and South America regularly visit so called salt or mineral licks to consume clay or drink clay-saturated water. Whether this behavior is used to supplement diets with locally limited nutrients or to buffer the effects of toxic secondary plant compounds remains unclear. In the Amazonian rainforest, pregnant and lactating bats are frequently observed and captured at mineral licks. We measured the nitrogen isotope ratio in wing tissue of omnivorous short-tailed fruit bats, Carollia perspicillata, and in an obligate fruit-eating bat, Artibeus obscurus, captured at mineral licks and at control sites in the rainforest. Carollia perspicillata with a plant-dominated diet were more often captured at mineral licks than individuals with an insect-dominated diet, although insects were more mineral depleted than fruits. In contrast, nitrogen isotope ratios of A. obscurus did not differ between individuals captured at mineral lick versus control sites. We conclude that pregnant and lactating fruit-eating bats do not visit mineral licks principally for minerals, but instead to buffer the effects of secondary plant compounds that they ingest in large quantities during periods of high energy demand. These findings have potential implications for the role of mineral licks for mammals in general, including humans. PMID- 18431490 TI - Etoposide induces ATM-dependent mitochondrial biogenesis through AMPK activation. AB - BACKGROUND: DNA damage such as double-stranded DNA breaks (DSBs) has been reported to stimulate mitochondrial biogenesis. However, the underlying mechanism is poorly understood. The major player in response to DSBs is ATM (ataxia telangiectasia mutated). Upon sensing DSBs, ATM is activated through autophosphorylation and phosphorylates a number of substrates for DNA repair, cell cycle regulation and apoptosis. ATM has been reported to phosphorylate the alpha subunit of AMP-activated protein kinase (AMPK), which senses AMP/ATP ratio in cells, and can be activated by upstream kinases. Here we provide evidence for a novel role of ATM in mitochondrial biogenesis through AMPK activation in response to etoposide-induced DNA damage. METHODOLOGY/PRINCIPAL FINDINGS: Three pairs of human ATM+ and ATM- cells were employed. Cells treated with etoposide exhibited an ATM-dependent increase in mitochondrial mass as measured by 10-N Nonyl-Acridine Orange and MitoTracker Green FM staining, as well as an increase in mitochondrial DNA content. In addition, the expression of several known mitochondrial biogenesis regulators such as the major mitochondrial transcription factor NRF-1, PGC-1alpha and TFAM was also elevated in response to etoposide treatment as monitored by RT-PCR. Three pieces of evidence suggest that etoposide induced mitochondrial biogenesis is due to ATM-dependent activation of AMPK. First, etoposide induced ATM-dependent phosphorylation of AMPK alpha subunit at Thr172, indicative of AMPK activation. Second, inhibition of AMPK blocked etoposide-induced mitochondrial biogenesis. Third, activation of AMPK by AICAR (an AMP analogue) stimulated mitochondrial biogenesis in an ATM-dependent manner, suggesting that ATM may be an upstream kinase of AMPK in the mitochondrial biogenesis pathway. CONCLUSIONS/SIGNIFICANCE: These results suggest that activation of ATM by etoposide can lead to mitochondrial biogenesis through AMPK activation. We propose that ATM-dependent mitochondrial biogenesis may play a role in DNA damage response and ROS regulation, and that defect in ATM-dependent mitochondrial biogenesis could contribute to the manifestations of A-T disease. PMID- 18431493 TI - Infection of cultured human endothelial cells by Legionella pneumophila. AB - Legionella pneumophila is a gram-negative pathogen that causes a severe pneumonia known as Legionnaires' disease. Here, we demonstrate for the first time that L. pneumophila infects and grows within cultured human endothelial cells. Endothelial infection may contribute to lung damage observed during Legionnaires' disease and to systemic spread of this organism. PMID- 18431494 TI - Mitochondrial membrane potential in human neutrophils is maintained by complex III activity in the absence of supercomplex organisation. AB - BACKGROUND: Neutrophils depend mainly on glycolysis for their energy provision. Their mitochondria maintain a membrane potential (Deltapsi(m)), which is usually generated by the respiratory chain complexes. We investigated the source of Deltapsi(m) in neutrophils, as compared to peripheral blood mononuclear leukocytes and HL-60 cells, and whether neutrophils can still utilise this Deltapsi(m) for the generation of ATP. METHODS AND PRINCIPAL FINDINGS: Individual activity of the oxidative phosphorylation complexes was significantly reduced in neutrophils, except for complex II and V, but Deltapsi(m) was still decreased by inhibition of complex III, confirming the role of the respiratory chain in maintaining Deltapsi(m). Complex V did not maintain Deltapsi(m) by consumption of ATP, as has previously been suggested for eosinophils. We show that complex III in neutrophil mitochondria can receive electrons from glycolysis via the glycerol 3-phosphate shuttle. Furthermore, respiratory supercomplexes, which contribute to efficient coupling of the respiratory chain to ATP synthesis, were lacking in neutrophil mitochondria. When HL-60 cells were differentiated to neutrophil-like cells, they lost mitochondrial supercomplex organisation while gaining increased aerobic glycolysis, just like neutrophils. CONCLUSIONS: We show that neutrophils can maintain Deltapsi(m) via the glycerol-3-phosphate shuttle, whereby their mitochondria play an important role in the regulation of aerobic glycolysis, rather than producing energy themselves. This peculiar mitochondrial phenotype is acquired during differentiation from myeloid precursors. PMID- 18431495 TI - Ndel1 promotes axon regeneration via intermediate filaments. AB - Failure of axons to regenerate following acute or chronic neuronal injury is attributed to both the inhibitory glial environment and deficient intrinsic ability to re-grow. However, the underlying mechanisms of the latter remain unclear. In this study, we have investigated the role of the mammalian homologue of aspergillus nidulans NudE, Ndel1, emergently viewed as an integrator of the cytoskeleton, in axon regeneration. Ndel1 was synthesized de novo and upregulated in crushed and transected sciatic nerve axons, and, upon injury, was strongly associated with neuronal form of the intermediate filament (IF) Vimentin while dissociating from the mature neuronal IF (Neurofilament) light chain NF-L. Consistent with a role for Ndel1 in the conditioning lesion-induced neurite outgrowth of Dorsal Root Ganglion (DRG) neurons, the long lasting in vivo formation of the neuronal Ndel1/Vimentin complex was associated with robust axon regeneration. Furthermore, local silencing of Ndel1 in transected axons by siRNA severely reduced the extent of regeneration in vivo. Thus, Ndel1 promotes axonal regeneration; activating this endogenous repair mechanism may enhance neuroregeneration during acute and chronic axonal degeneration. PMID- 18431497 TI - A test of highly optimized tolerance reveals fragile cell-cycle mechanisms are molecular targets in clinical cancer trials. AB - Robustness, a long-recognized property of living systems, allows function in the face of uncertainty while fragility, i.e., extreme sensitivity, can potentially lead to catastrophic failure following seemingly innocuous perturbations. Carlson and Doyle hypothesized that highly-evolved networks, e.g., those involved in cell cycle regulation, can be resistant to some perturbations while highly sensitive to others. The "robust yet fragile" duality of networks has been termed Highly Optimized Tolerance (HOT) and has been the basis of new lines of inquiry in computational and experimental biology. In this study, we tested the working hypothesis that cell-cycle control architectures obey the HOT paradigm. Three cell-cycle models were analyzed using monte-carlo sensitivity analysis. Overall state sensitivity coefficients, which quantify the robustness or fragility of a given mechanism, were calculated using a monte-carlo strategy with three different numerical techniques along with multiple parameter perturbation strategies to control for possible numerical and sampling artifacts. Approximately 65% of the mechanisms in the G1/S restriction point were responsible for 95% of the sensitivity, conversely, the G2-DNA damage checkpoint showed a much stronger dependence on a few mechanisms; approximately 32% or 13 of 40 mechanisms accounted for 95% of the sensitivity. Our analysis predicted that CDC25 and cyclin E mechanisms were strongly implicated in G1/S malfunctions, while fragility in the G2/M checkpoint was predicted to be associated with the regulation of the cyclin B-CDK1 complex. Analysis of a third model containing both G1/S and G2/M checkpoint logic, predicted in addition to mechanisms already mentioned, that translation and programmed proteolysis were also key fragile subsystems. Comparison of the predicted fragile mechanisms with literature and current preclinical and clinical trials suggested a strong correlation between efficacy and fragility. Thus, when taken together, these results support the working hypothesis that cell-cycle control architectures are HOT networks and establish the mathematical estimation and subsequent therapeutic exploitation of fragile mechanisms as a novel strategy for anti-cancer lead generation. PMID- 18431496 TI - Comparative genomic analysis of clinical strains of Campylobacter jejuni from South Africa. AB - BACKGROUND: Campylobacter jejuni is a common cause of acute gastroenteritis and is also associated with the post-infectious neuropathies, Guillain-Barre and Miller Fisher syndromes. In the Cape Town area of South Africa, C. jejuni strains with Penner heat-stable (HS) serotype HS:41 have been observed to be overrepresented among cases of Guillain-Barre syndrome. The present study examined the genetic content of a collection of 32 South African C. jejuni strains with different serotypes, including 13 HS:41 strains, that were recovered from patients with enteritis, Guillain-Barre or Miller Fisher syndromes. The sequence-based typing methods, multilocus sequence typing and DNA microarrays, were employed to potentially identify distinguishing features within the genomes of these C. jejuni strains with various disease outcomes. METHODOLOGY/PRINCIPAL FINDINGS: Comparative genomic analyses demonstrated that the HS:41 South African strains were clearly distinct from the other South African strains. Further DNA microarray analysis demonstrated that the HS:41 strains from South African patients with the Guillain-Barre syndrome or enteritis were highly similar in gene content. Interestingly, the South African HS:41 strains were distinct in gene content when compared to HS:41 strains from other geographical locations due to the presence of genomic islands, referred to as Campylobacter jejuni integrated elements (CJIEs). Only the integrated element CJIE1, a Campylobacter Mu-like prophage, was present in the South African HS:41 strains whereas this element was absent in two closely-related HS:41 strains from Mexico. A more distantly-related HS:41 strain from Canada possessed both integrated elements CJIE1 and CJIE2. CONCLUSION/SIGNIFICANCE: These findings demonstrate that CJIEs may contribute to the differentiation of closely-related C. jejuni strains. In addition, the presence of bacteriophage-related genes in CJIE1 may contribute to the genomic diversity of C. jejuni strains. This comparative genomic analysis of C. jejuni provides fundamental information that potentially could lead to improved methods for analyzing the epidemiology of disease outbreaks. PMID- 18431498 TI - Metabolic and immune activation effects of treatment interruption in chronic HIV 1 infection: implications for cardiovascular risk. AB - BACKGROUND: Concern about costs and antiretroviral therapy (ART)-associated toxicities led to the consideration of CD4 driven strategies for the management of HIV. That approach was evaluated in the SMART trial that reported an unexpected increase of cardiovascular events after treatment interruption (TI). Our goal was to evaluate fasting metabolic changes associated with interruption of antiretroviral therapy and relate them to changes of immune activation markers and cardiovascular risk. METHODOLOGY: ACTG 5102 enrolled 47 HIV-1-infected subjects on stable ART, with <200 HIV RNA copies/mL and CD4 cell count >or=500 cells/microL. Subjects were randomly assigned to continue ART for 18 weeks with or without 3 cycles of interleukin-2 (IL-2) (cycle = 4.5 million IU sc BID x 5 days every 8 weeks). After 18 weeks ART was discontinued in all subjects until the CD4 cell count dropped below 350 cells/microL. Glucose and lipid parameters were evaluated every 8 weeks initially and at weeks 2, 4, 8 and every 8 weeks after TI. Immune activation was evaluated by flow-cytometry and soluble TNFR2 levels. PRINCIPAL FINDINGS: By week 8 of TI, levels of total cholesterol (TC) (median (Q1, Q3) (-0.73 (-1.19, -0.18) mmol/L, p<0.0001), LDL, HDL cholesterol ( 0.36(-0.73,-0.03)mmol/L, p = 0.0007 and -0.05(-0.26,0.03), p = 0.0033, respectively) and triglycerides decreased (-0.40 (-0.84, 0.07) mmol/L, p = 0.005). However the TC/HDL ratio remained unchanged (-0.09 (-1.2, 0.5), p = 0.2). Glucose and insulin levels did not change (p = 0.6 and 0.8, respectively). After TI there was marked increase in immune activation (CD8+/HLA-DR+/CD38+ cells, 34% (13, 43), p<0.0001) and soluble TNFR2 (1089 ng/L (-189, 1655), p = 0.0008) coinciding with the rebound of HIV viremia. CONCLUSIONS: Our data suggests that interrupting antiretroviral therapy does not reduce cardiovascular disease (CVD) risk, as the improvements in lipid parameters are modest and overshadowed by the decreased HDL levels. Increased immune cell activation and systemic inflammatory responses associated with recrudescent HIV viremia may provide a more cogent explanation for the increased cardiovascular risk associated with treatment interruption and HIV infection. TRIAL REGISTRATION: ClinicalTrials.gov NCT00015704. PMID- 18431499 TI - The septins function in G1 pathways that influence the pattern of cell growth in budding yeast. AB - The septins are a conserved family of proteins that have been proposed to carry out diverse functions. In budding yeast, the septins become localized to the site of bud emergence in G1 but have not been thought to carry out important functions at this stage of the cell cycle. We show here that the septins function in redundant mechanisms that are required for formation of the bud neck and for the normal pattern of cell growth early in the cell cycle. The Shs1 septin shows strong genetic interactions with G1 cyclins and is directly phosphorylated by G1 cyclin-dependent kinases, consistent with a role in early cell cycle events. However, Shs1 phosphorylation site mutants do not show genetic interactions with the G1 cyclins or obvious defects early in the cell cycle. Rather, they cause an increased cell size and aberrant cell morphology that are dependent upon inhibitory phosphorylation of Cdk1 at the G2/M transition. Shs1 phosphorylation mutants also show defects in interaction with the Gin4 kinase, which associates with the septins during G2/M and plays a role in regulating inhibitory phosphorylation of Cdk1. Phosphorylation of Shs1 by G1 cyclin-dependent kinases plays a role in events that influence Cdk1 inhibitory phosphorylation. PMID- 18431500 TI - Cooperation and deception recruit different subsets of the theory-of-mind network. AB - The term "theory of mind" (ToM) describes an evolved psychological mechanism that is necessary to represent intentions and expectations in social interaction. It is thus involved in determining the proclivity of others to cooperate or defect. While in cooperative settings between two parties the intentions and expectations of the protagonists match, they diverge in deceptive scenarios, in which one protagonist is intentionally manipulated to hold a false belief about the intention of the other. In a functional magnetic resonance imaging paradigm using cartoons showing social interactions (including the outcome of the interaction) between two or three story characters, respectively, we sought to determine those brain areas of the ToM network involved in reasoning about cooperative versus deceptive interactions. Healthy volunteers were asked to reflect upon the protagonists' intentions and expectations in cartoons depicting cooperation, deception or a combination of both, where two characters cooperated to deceive a third. Reasoning about the mental states of the story characters yielded substantial differences in activation patterns: both deception and cooperation activated bilateral temporoparietal junction, parietal and cingulate regions, while deception alone additionally recruited orbitofrontal and medial prefrontal regions. These results indicate an important role for prefrontal cortex in processing a mismatch between a character's intention and another's expectations as required in complex social interactions. PMID- 18431501 TI - BRCA1 and BRCA2 germline mutations in Malaysian women with early-onset breast cancer without a family history. AB - BACKGROUND: In Asia, breast cancer is characterised by an early age of onset: In Malaysia, approximately 50% of cases occur in women under the age of 50 years. A proportion of these cases may be attributable, at least in part, to genetic components, but to date, the contribution of genetic components to breast cancer in many of Malaysia's ethnic groups has not been well-characterised. METHODOLOGY: Given that hereditary breast carcinoma is primarily due to germline mutations in one of two breast cancer susceptibility genes, BRCA1 and BRCA2, we have characterised the spectrum of BRCA mutations in a cohort of 37 individuals with early-onset disease ( 22% was the best predictor of a >25% reduction of Ac with 76% sensitivity and 65% specificity (area 71%, CI 54%-85%, P = 0.02). In addition an >32% increase of IL-6 at 120 min of reperfusion predicted a >25% reduction of Ac with a 76% sensitivity and 65% specificity (area 71% CI 61%-97%, P = 0.01). CONCLUSION: Changes of mean diastolic CF and IL-6 cs are associated with alterations in myocardial microvascular integrity after ischaemia-reperfusion and may be used as a predictor of myocardial dysfunction. PMID- 18431528 TI - Anterior pedicle screw fixation for multilevel cervical corpectomy and spinal fusion. AB - BACKGROUND: Prevention of graft dislodgement in multilevel cervical corpectomy and fusion has been an unresolved problem. Anterior plate fixation has a significant failure rate. External support with a halo-vest is uncomfortable for patients. In the present study, we report a new surgical technique of anterior pedicle screw (APS) fixation for multilevel cervical corpectomy and spinal fusion, and describe the safety and utility of the system. METHOD: After cervical corpectomy, the pedicles on the right side were visualised under oblique fluoroscopy. Guide wires were inserted into the pedicles from the inner wall of the excavated vertebral body until they were hidden in the pedicles. After a fibula autograft was placed, the graft was penetrated in the reverse direction by the guide wires. After drilling and tapping, cannulated screws were inserted into the pedicles through the grafted fibula along the guide wires. FINDINGS: In 9 patients with cervical myelopathy, the surgery was accomplished with a fibula autograft using APS fixation. A total of 22 APSs were inserted, and 21 screws were placed precisely in the pedicles. There were no neurovascular complications. Patients were allowed to ambulate without a halo-vest on the second day after the surgery. Post-operatively, no dislodgement of the grated fibula occurred, and all patients improved neurologically. CONCLUSIONS: The insertion of APSs is feasible and safe. APS fixation enables us to obtain rigid fixation anteriorly, and we propose that APS fixation is an attractive option for multilevel cervical corpectomy and fusion. PMID- 18431529 TI - Trigeminal fistula treated by combined transvenous and transarterial embolisation. AB - Trigeminal cavernous fistulae are rare, compared to other types of carotid cavernous fistulae with only about 17 examples previously reported in English literature. The aetiology can be either post-traumatic or spontaneous, and in the latter group either due to a ruptured trigeminal aneurysm or postulated intrinsic weakness of the artery itself. The treatment goal is to close the fistula while preserving the parent artery and this can be done either by surgical or endovascular methods. We present a 35-year-old woman with a spontaneous trigeminal fistula treated by combined trans-venous and trans-arterial embolisation. PMID- 18431530 TI - Reversible deficit affecting the perception of tone of a human voice after tumour resection from the right auditory cortex. AB - We report on a young woman operated for a ganglioglioma involving the right auditory cortex (AC), presenting with auditory seizures. Despite a normal pre operative examination, a specific post-operative disorder affecting the perception of a human voice occurred. The patient was unable to recognise the tone of familiar voices while she recognised the expressed content. A temporal lobectomy for recurrence was performed two years later. The patient recovered from the voice perception deficit. This report shows that (1) a discrete site within the AC is specifically involved in the perception of tone of the human voice (2) functional compensation is possible. PMID- 18431531 TI - Use of tetrapeptide signals for protein secondary-structure prediction. AB - This paper develops a novel sequence-based method, tetra-peptide-based increment of diversity with quadratic discriminant analysis (TPIDQD for short), for protein secondary-structure prediction. The proposed TPIDQD method is based on tetra peptide signals and is used to predict the structure of the central residue of a sequence fragment. The three-state overall per-residue accuracy (Q (3)) is about 80% in the threefold cross-validated test for 21-residue fragments in the CB513 dataset. The accuracy can be further improved by taking long-range sequence information (fragments of more than 21 residues) into account in prediction. The results show the tetra-peptide signals can indeed reflect some relationship between an amino acid's sequence and its secondary structure, indicating the importance of tetra-peptide signals as the protein folding code in the protein structure prediction. PMID- 18431532 TI - Analysis of 3D structural differences in the IgG-binding domains based on the interresidue average-distance statistics. AB - It is well-known that the IgG-binding domain from staphylococcal protein A folds into a 3 alpha helix bundle structure, while the IgG-binding domain of streptococcal protein G forms an (alpha + beta) structure. Recently, He et al. (Biochemistry 44:14055-14061, 2005) made mutants of these proteins from the wild types of protein A and protein G strains. These mutants are referred to as protein A219 and protein G311, and it was showed that these two mutants have different 3D structures, i.e., the 3 alpha helix bundle structure and the (alpha + beta) structure, respectively, despite the high sequence identity (59%). The purpose of our study was to clarify how such 3D structural differences are coded in the sequences with high homology. To address this problem, we introduce a predicted contact map constructed based on the interresidue average-distance statistics for prediction of folding properties of a protein. We refer to this map as an average distance map (ADM). Furthermore, the statistics of interresidue distances can be converted to an effective interresidue potential. We calculated the contact frequency of each residue of a protein in random conformations with this effective interresidue potential, and then we obtained values similar to phi values. We refer to this contact frequency of each residue as a p(mu) value. The comparison of the p(mu) values to the phi values for a protein suggests that p(mu) values reveal the information on the folding initiation site. Using these techniques, we try to extract the information on the difference in the 3D structures of protein A219 and protein G311 coded in their amino acid sequences in the present work. The results show that the ADM analyses and the p(mu) value analyses predict the information of folding initiation sites, which can be used to detect the 3D difference in both proteins. PMID- 18431533 TI - The importance of proline residues in the structure, stability and susceptibility to proteolytic degradation of collagens. AB - Collagens are among proteins that undergo several post-translational modifications, such as prolyl hydroxylation, that occur during elongation of the nascent chains in the endoplasmic reticulum. The major structural collagens, types I, II and III, have large, uninterrupted triple helices, comprising three polyproline II-like chains supercoiled around a common axis. The structure has a requirement for glycine, as every third residue, and is stabilized by the high content of proline and 4-hydroxyproline residues. Action of prolyl hydroxylases is critical. Spontaneous or targeted genetic defects in prolyl hydroxylases can be lethal or result in severe osteogenesis imperfecta. Prolines, as determinants of substrate specificity and susceptibility, also play a role in degradation of collagen by collagenolytic matrix metalloproteinases (MMPs). Targeted mutations in mice in the collagenase cleavage domain have profound effects on collagen turnover and the function of connective tissues. Prolines are thus critical determinants of collagen structure and function. PMID- 18431534 TI - A randomised controlled trial into the effects of food on ADHD. AB - The aim of this study is to assess the efficacy of a restricted elimination diet in reducing symptoms in an unselected group of children with Attention deficit/hyperactivity disorder (ADHD). Dietary studies have already shown evidence of efficacy in selected subgroups. Twenty-seven children (mean age 6.2) who all met the DSM-IV criteria for ADHD, were assigned randomly to either an intervention group (15/27) or a waiting-list control group (12/27). Primary endpoint was the clinical response, i.e. a decrease in the symptom scores by 50% or more, at week 9 based on parent and teacher ratings on the abbreviated ten item Conners Scale and the ADHD-DSM-IV Rating Scale. The intention-to-treat analysis showed that the number of clinical responders in the intervention group was significantly larger than that in the control group [parent ratings 11/15 (73%) versus 0/12 (0%); teacher ratings, 7/10 (70%) versus 0/7 (0%)]. The Number of ADHD criteria on the ADHD Rating Scale showed an effect size of 2.1 (cohen's d) and a scale reduction of 69.4%. Comorbid symptoms of oppositional defiant disorder also showed a significantly greater decrease in the intervention group than it did in the control group (cohens's d 1.1, scale reduction 45.3%). A strictly supervised elimination diet may be a valuable instrument in testing young children with ADHD on whether dietary factors may contribute to the manifestation of the disorder and may have a beneficial effect on the children's behaviour. PMID- 18431535 TI - Psychopathology and prosocial behavior in adolescents from socio-economically disadvantaged families: the role of proximal and distal adverse life events. AB - The study investigated if proximal contextual risk (number of adverse life events experienced in the last year) or distal contextual risk (number of adverse life events experienced before the last year) is a better predictor of adolescent psychopathology and prosocial behavior. It also tested for the specificity, accumulation and gradient of contextual risk in psychopathology and prosocial behavior, and for the interaction between proximal and distal contextual risk in psychopathology and prosocial behavior. The sample was 199 11-18 year old children from a socio-economically disadvantaged area in North-East London. The Strengths and Difficulties Questionnaire (SDQ), which measures four difficulties (hyperactivity, emotional symptoms, conduct problems, and peer problems) and prosocial behavior, was used. Confounders were age, gender, and maternal educational qualifications. To model the relationship between the five SDQ scales and contextual risk multivariate response regression models and multivariate response logistic regression models that allow the error terms of the scale specific models to be correlated were fitted. This study highlighted the importance of proximal contextual risk in predicting both broad and externalizing psychopathology, and the importance of considering risk accumulation rather than specificity in predicting psychopathology. By showing that the number of proximal adverse life events experienced had a steady, additive effect on broad and externalizing psychopathology, it also highlighted the need to protect adolescents experiencing current risk from further risk exposure. By showing that the number of distal adverse life events experienced did not affect the proximal risk's impact on either broad or externalizing psychopathology, it highlighted the need to protect all adolescents, irrespective of experience of early life adversities, from risk. PMID- 18431536 TI - Parent and child agreement on reports of problem behaviour obtained from a screening questionnaire, the SDQ. AB - BACKGROUND AND OBJECTIVES: This study examined the level of agreement between parents and children on the Strengths and Difficulties Questionnaire (SDQ) in a clinical sample in Sydney, Australia. METHODS: Parent and child SDQ reports were collected from 379 parents-child pairs. Children were aged between 11 and 18 years and met criteria for at least one psychiatric diagnosis. RESULTS: Overall agreement between children and parents was low to modest. Sixty nine percent of parent-child pairs agreed that the child's problems were either clinically significant or not ("neither", "both"), while in 27% of pairs only the parents regarded the problems as clinically significant ("parent only"), and in 4% of pairs only the children regarded the problems as clinically significant. There was higher agreement for children with mood, anxiety or somatoform disorders. Children with mood disorders were over-represented in the "child only" group, and those with conduct disorders were more likely to be in the "parent only" group. Children with anxiety and somatoform disorders were more likely to be in the "neither" group. Age was not associated with rates of parent-child agreement, however more girls agreed with their parents that either they did not have a problem ("neither") or they did have a problem without parental endorsement ("child only"). CONCLUSIONS: This study highlights the limited agreement between parent and child reports of problem behaviour and the importance of integrating discrepant information into child and adolescent mental health assessments, formulations and treatments. PMID- 18431537 TI - Do larger studies find smaller effects? The example of studies for the prevention of conduct disorder. AB - INTRODUCTION: There is some emerging evidence in medicine that larger clinical trials tend to be associated with smaller effect sizes. Much of the evidence-base currently informing practice in Child Psychiatry relies on relatively small trials. We therefore investigated the relationship between trial size and effect size in research within a key area of child mental health. METHOD: A recent systematic review of 20 trials of prevention of conduct disorder was subjected to meta-regression, to examine the relationship between study size and effect size, and to explore hypothesised confounding variables. RESULTS: In this sample of studies, reported effect size was inversely related to sample size. This effect is not explained by year of publication, intervention type or quality of methodology in the study. DISCUSSION: Our finding is consistent with other reports in the literature. The origin of this effect is not yet clear. However if replicated it clearly has significant implications for the way trials in child mental health are interpreted. PMID- 18431538 TI - A bibliometric study of international scientific productivity in attention deficit hyperactivity disorder covering the period 1980-2005. AB - OBJECTIVE: We have carried out a bibliometric analysis of scientific publications related to attention-deficit hyperactivity disorder (ADHD) and its pharmacological treatment over the period 1980-2005. METHOD: We selected (in EMBASE and MEDLINE databases) documents that contained in their title the descriptors attention deficit hyperactivity disorder, attention deficit disorder, ADHD and ADD. As bibliometric indicators of production and dispersion we applied Price's Law and Bradford's Law, respectively. We also calculated the national participation index (PI) and correlated it with overall PI in biomedical and health sciences, with PI in the discipline of Psychiatry and with the social health indicators (per capita Gross Domestic Product, number of physicians and total per capita expenditure on health). RESULTS: We obtained 5,269 original documents (2,325 corresponded to pharmacological therapy). Our results indicate fulfilment of Price's Law, since scientific production on ADHD undergoes exponential growth (correlation coefficient r = 0.9859, vs. r = 0.9011 after linear adjustment). The most widely studied drugs are methylphenidate (1,251 documents). Division into Bradford zones yields a nucleus occupied exclusively by the Journal of the American Academy of Child and Adolescent Psychiatry (500 articles). A total of 866 different journals were employed. Twelve of the first 20 used journals have an Impact Factor > 2. The principal producer country is the United States (PI = 44.2). Only four countries, of the 20 major producers in health sciences, surpass their own PI in the field of Psychiatry (Brazil, China, Spain and the United States). The correlation between PI and total number of physicians for each country situates Canada, Australia, the United States and Israel in the top positions. CONCLUSION: Productivity on ADHD has undergone exponential growth in the period 1980-2005, without evidence a saturation point. PMID- 18431539 TI - What single reports from children and parents aggregate to attention deficit hyperactivity disorder and oppositional defiant disorder diagnoses in epidemiological studies. AB - OBJECTIVE: To analyze information on attention deficit-hyperactivity disorder (ADHD)-oppositional defiant disorder (ODD) and its consequences, provided separately or in combination by children and their parents in a longitudinal prospective study of 9-15 year-old children from the general population. METHOD: Cross-sectional and longitudinal epidemiological indexes were compared for single and multiple reports. We evaluated which informant is required for the identification of each DSM-IV criterion. Logistic regressions determined which features were related with the reporting of the "absence" of symptoms. RESULTS: Both informants were required in order to obtain complete psychopathological profiles. Single reports provide infra-estimated prevalences (between 8.8 and 22.9% of ADHD and between 1.7 and 7.6% of ODD), risks (around 3% for ADHD and 2% for ODD) and comorbidities. Psychological and functional measures analyzed in the study were relatively similar for cases presenting ADHD/ODD diagnosis, regardless of the diagnostic algorithm (based on single or combined reports); however, these clinical profiles were different to those obtained for non-diagnosed children. The main predictors of not reporting the presence of psychopathology were: large families (OR between 2 and 2.5), children that are conflictive at school (OR ranging between 1.3 and 4.3) or those with poor mental health (OR between 1.1 and 1.6). CONCLUSIONS: These results may provide guidance for obtaining accurate diagnostic information, properly identifying children with mental health needs and planning the required preventive and corrective measures. PMID- 18431540 TI - Prevalence and correlates of conduct disorder and problem behavior in Caribbean and Filipino immigrant adolescents. AB - This study investigates the prevalence and subtypes of conduct disorder (CD) and behavioral problems among youth in two communities characterized by prolonged parent-child separation upon immigration. CD and problem behaviors were assessed in 252 Caribbean-Canadian and Filipino-Canadian adolescents (12-19-year-old) using the DISC-C, the YSR and the CBCL cross-informant construct. Adolescents reported less problem behaviors than their host country peers, despite immigrant background or parent-child separation. The high adolescent-onset CD rate supports the hypothesis that psychosocial stressors play a role in the emergence of the disorder. Specifically, high levels of perceived racism and low collective self esteem predicted problem behaviors in these youngsters. PMID- 18431541 TI - The efficacy of intermittent cervical traction in patents with chronic neck pain. AB - Previous studies about the usefulness of traction therapy have concluded with conflicting results. The aim of this study was to examine its efficacy in chronic neck pain. Forty-two patients with at least 6 weeks of nonspecific neck pain were selected for the study. Data about demographic characteristics including age, sex, body mass index, duration of cervical pain, working status, smoking status, and regular exercise were recorded. Each patient was randomly assigned to Group 1 receiving only standard physical therapy including hot pack, ultrasound therapy and exercise program and Group 2-treated with traction therapy in addition to standard physical therapy. The patients were reevaluated at the end of the therapy. The main outcome measures of the treatment were pain intensity by visual analog scale (VAS), disability by neck disability index (NDI), and quality of life assessed by Nottingham Health Profile (NHP). Twenty-four female and 18 male patients with mean age of 48.2+/-11.5 years and a mean disease duration of 4.3+/ 2.9 years were included to the study. There were no differences between the groups in terms of age, sex, pain intensity, and scores of NHP and NDI at entry. There were 21 patients in both groups. Both groups improved significantly in pain intensity and the scores of NDI and physical subscles of NHP at the end of the therapies (p<0.05). There was an association between NDI and VAS pain scores in both groups (p<0.05). No correlation was observed between clinical variables and age and duration of disease. In conclusion, no specific effect of traction over standard physiotherapeutic interventions was observed in adults with chronic neck pain. We suggest the clinicians to consider this condition and to focus on exercise therapy in the management of patients suffering from this condition. PMID- 18431542 TI - Wetting transitions on textured hydrophilic surfaces. AB - We consider the quasi-static energy of a drop on a textured hydrophilic surface, with taking the contact angle hysteresis (CAH) into account. We demonstrate how energy varies as the contact state changes from the Cassie state (in which air is trapped at the drop bottom) to the Wenzel state (in which liquid fills the texture at the drop bottom) assuming that the latter state nucleates from the center of the drop bottom. When the textured substrate is hydrophilic enough to allow spontaneous penetration of liquid film of the texture thickness, the present theory asserts that the drop develops into an experimentally observed state in which a drop looks like an egg fried without flipped over (sunny-side up) with a well-defined radius of "the egg yolk." Otherwise, the final contact state of the drop becomes like a Wenzel state, but with the contact circle smaller than the original Wenzel state due to the CAH. We provide simple analytical estimations for the yolk radius of the "sunny-side-up" state and for the final radius of the contact circle of the pseudo-Wenzel state. PMID- 18431543 TI - Expression of bcl-2 is associated with microvessel density in olfactory neuroblastoma. AB - Erythropoietin (Epo) expression is regulated via hypoxia-inducible factor (HIF) 1alpha-directed gene transcription. Activation of the erythropoietin receptor (EpoR) by Epo leads to elevated expression of the anti-apoptotic protein, bcl-2, which has recently been shown to promote angiogenesis in malignant tumors. Expression of HIF-1alpha, Epo, EpoR, and bcl-2 was studied by immunohistochemistry in a series of 20 olfactory neuroblastoma (ONB) samples. Data were correlated with microvessel density, proliferative activity, and apoptosis in the specimens and survival analysis was performed to investigate the prognostic value of the examined factors. Immunohistochemical analysis revealed robust expression of HIF-1alpha, Epo, EpoR, and bcl-2 in ONB. Ninety percent of the samples showed HIF-1alpha immunoreactivity and in 60% of the cases, bcl-2 immunoreactivity was observed. A significant positive correlation between the expression levels of HIF-1alpha and bcl-2 and the microvessel density was found. Survival analysis did not reveal any prognostic significance for the tested factors. Expression of HIF-1alpha, Epo, Epo-R, and bcl-2 may play a functional role in ONB pathogenesis. Our data suggest that bcl-2 may act as a stimulator of angiogenesis in ONB, and thus represents a novel target for anti-angiogenic treatment strategies in the therapy of ONB. PMID- 18431545 TI - A comparison of EEOC closures involving hiring versus other prevalent discrimination issues under the Americans with Disabilities Act. AB - INTRODUCTION: This article describes findings from a causal comparative study of the Merit Resolution rate for allegations of Hiring discrimination that were filed with the U.S. Equal Employment Opportunity Commission (EEOC) under Title I of the Americans with Disabilities Act (ADA) between 1992 and 2005. An allegation is the Charging Party's perception of discrimination, but a Merit Resolution is one in which the EEOC has determined that a discriminatory event did indeed occur. A Non-Merit Resolution is an allegation that is closed due to a technicality or lacks sufficient evidence to conclude that discrimination occurred. Merit favors the Charging Party; Non-Merit favors the Employer. METHODS: The Merit Resolution rate of 19,527 closed Hiring allegations is compared and contrasted to that of 259,680 allegations aggregated from six other prevalent forms of discrimination including Discharge and Constructive Discharge, Reasonable Accommodation, Disability Harassment and Intimidation, and Terms and Conditions of Employment. Tests of Proportion distributed as chi-square are used to form comparisons along a variety of subcategories of Merit and Non-Merit outcomes. RESULTS: The overall Merit Resolution rate for Hiring is 26% compared to Non-Hiring at 20.6%. Employers are less likely to settle claims of hiring discrimination without mediation, and less likely to accept the remedies recommended by the EEOC when hiring discrimination has been determined. CONCLUSION: Hiring is not an unusual discrimination issue in that the overwhelming majority of allegations are still closed in favor of the Employer. However, it is counterintuitive that hiring has a higher merit resolution rate than other prevalent issues. This finding contradicts the assumption that hiring is an "invisible process." Considering that the EEOC makes merit determinations at a competitive rate, it is clear that hiring is sufficiently transparent. PMID- 18431544 TI - Temozolomide three weeks on and one week off as first line therapy for patients with recurrent or progressive low grade gliomas. AB - BACKGROUND: Patients with recurrent or progressive low grade gliomas survive for a decade or more following diagnosis, and may be at a higher risk for treatment related complications, such as cognitive impairment from radiotherapy. PURPOSE: The aim of the present study was to determine in patients with progressive or recurrent low grade gliomas, the response rate and toxicity incurred by a continued schedule of temozolomide chemotherapy administered before radiation therapy, and to explore correlations between response and survival with 1p/19q deletions and MGMT promoter methylation status. METHODS: Progressive radio and chemotherapy naive low grade glioma patients with O(6)-methyl-guanine-DNA-methyl tranferase (MGMT) promoter status evaluation were considered eligible. Chemotherapy cycles consisted of temozolomide 75 mg/m(2)/daily for 21 days every 28 days for 12 cycles. RESULTS: A total of 30 patients (median age 45 [range: 24.2-68.6] years) with a median KPS of 90 (range 60-90) were accrued. The overall response rate was 30% (9 partial responses); 17 patients (56.7%) had disease stabilization. CONCLUSION: The prolonged temozolomide schedule considered in the present study is followed by a high response rate; toxicity is acceptable. Further randomized trials should therefore be conducted to confirm the efficacy of this regimen as first-line therapy in patients with progressive low grade glioma. PMID- 18431546 TI - Hiring discrimination against people with disabilities under the ADA: characteristics of employers. AB - INTRODUCTION: This article describes findings from a causal comparative study of the characteristics of employers against whom allegations of hiring discrimination were filed with the U.S. Equal Employment Opportunity Commission (EEOC) under Title I of the Americans with Disabilities Act (ADA) between 1992 and 2005. METHODS: Employer characteristics derived from 19,527 closed Hiring allegations are compared and contrasted to 259,680 closed allegations aggregated from six other prevalent forms of discrimination including Discharge and Constructive Discharge, Reasonable Accommodation, Disability Harassment and Intimidation, and Terms and Conditions of Employment. Tests of Proportion distributed as chi-square are used to form comparisons along a variety of factors including industry classification, size of workforce, and location. RESULTS: As compared to non-hiring allegations, hiring allegations were more likely to be filed against employers with 15-100 employees, in the West U.S. Census track region, or in industries including educational services; public administration; transportation and warehousing; professional, scientific, and technical services; agriculture, forestry, fishing, and hunting; and construction. CONCLUSION: More outreach regarding ADA responsibilities appears indicated for those employers who share the aforementioned characteristics. PMID- 18431547 TI - Hiring discrimination against people with disabilities under the ADA: characteristics of charging parties. AB - INTRODUCTION: This article describes findings from a causal comparative study of the characteristics of Charging Parties who filed allegations of Hiring discrimination with the U.S. Equal Employment Opportunity Commission (EEOC) under Title I of the Americans with Disabilities Act (ADA) between 1992 and 2005. METHODS: Charging Party Characteristics derived from 19,527 closed Hiring allegations are compared and contrasted to 259,680 closed allegations aggregated from six other prevalent forms of discrimination including Discharge and Constructive Discharge, Reasonable Accommodation, Disability Harassment and Intimidation, and Terms and Conditions of Employment. Tests of Proportion distributed as chi-square are used to form comparisons along a variety of factors including age, gender, impairment, and ethnicity. RESULTS: Most allegations of ADA job discrimination fall into the realm of job retention and career advancement as opposed to job acquisition. Hiring allegations, however, tend to be filed by Charging Parties who are disproportionately male, younger or older applicants, white, and coping with physical or sensory disabilities. CONCLUSION: Prevailing theories about stigma suggest that negative attitudes are more prevalent toward persons with behavioral disabilities. However, this study provides clear evidence that one behavioral manifestation of negative attitudes, Hiring discrimination, is more often directed at persons with physical or sensory impairments. More outreach regarding ADA rights appears indicated for individuals who share the aforementioned characteristics. PMID- 18431548 TI - Diffusion coefficients of several rhodamine derivatives as determined by pulsed field gradient-nuclear magnetic resonance and fluorescence correlation spectroscopy. AB - Rhodamine derivatives are popular, photostable fluorophores that are used in a number of fluorescent based techniques, including fluorescence correlation spectroscopy (FCS). Indeed, in FCS, both rhodamine 6G (R6G) and rhodamine 110 (R110) are used as calibration standards to determine the dimensions of the instrument confocal volume. In spite of a requirement for precise values of the diffusion coefficients, literature values are scarce and vary over an order of magnitude. In this paper, the diffusion coefficients of four rhodamine fluorophores (rhodamine 6G (R6G), rhodamine B (RB), rhodamine 123 (R123), rhodamine 110 (R110)) were determined by pulsed field gradient nuclear magnetic resonance (PFG-NMR) spectrometry and then validated by comparison with fluorescence correlation spectroscopy. With the objective of validating the FCS calibration, diffusion coefficients of several dextrans and a polystyrene nanoparticle were also determined and compared with literature values or theoretical values that were based upon the Stoke-Einstein equation. The work presented here lead us to conclude that the diffusion coefficients for R6G and R110 have generally been underestimated in the literature. We propose revised values of 4.4x10(-10) m2 s(-1) for R110 and 4.0x10(-10) m2 s(-1) for R6G. Using the revised D value for R110 to calibrate the FCS instrument, diffusion coefficients have then been systematically determined for different conditions of pH, ionic strength and concentration. To correct for differences due to solvent effects (D2O vs. H2O), an isotopic correction factor, DD2O/DH2O of 1.23, was determined from both FCS and from the solvent auto-diffusion coefficients obtained by NMR. PMID- 18431549 TI - ATP synthases: bioinformatic based insights into how their electrochemically driven motor comprised of subunits a and c might serve as a drug target. AB - ATP synthases, widely distributed in bacteria, eukaryotic mitochondria and chloroplasts, are highly conserved multi-subunit complexes. Although the conserved acidic residue in the transmembrane helix of the c subunit functions in H+ transport, the surrounding residues differ among species. Such divergence could lead to different regulatory modes since pH-dependent H+ transport has been demonstrated in E. coli with a c subunit carrying an additional acidic residue in the helix. There is further divergence in the number of c subunits that form the ring structure which is determined by the higher ordered structure. Recently, it was suggested that certain chemicals recognize the a and c subunits of pathogenic bacterial F0. Since there may be structural divergence even in well-conserved ATP synthases, the c subunit-ring as well as the a subunit in F0 could be targets for drugs for specific bacterial species. PMID- 18431551 TI - Hepatoprotective and antioxidant activity of methanol extract of Ficus glomerata. AB - The present study was carried out to evaluate Ficus glomerata extracts for antioxidant and hepatoprotective properties. The methanol extract of the bark of F. glomerata showed potent in vitro antioxidant activity when compared to the root methanol extract. In the in vivo studies, the CCl(4) treated control rats showed a significant alteration in the levels of antioxidant and hepatoprotective parameters. The methanol extract of the bark when given orally along with CCl(4) at the doses of 250 and 500 mg/kg body weight showed a significant reversal of these biochemical changes towards the normal when compared to CCl(4)-treated control rats in serum, liver and kidney. The results were comparable to those observed for standard sylimarin. Histological studies also confirmed the same. The results indicated the potent hepatoprotective and antioxidant nature of the bark extract. PMID- 18431552 TI - Quality of life one year after chemoradiotherapy for localized primary gastric diffuse large B-cell lymphoma. AB - We assessed the quality of life (QOL) at least one year after sequential chemoradiotherapy for the treatment of localized gastric diffuse large B-cell lymphoma (DLBCL). We used the EORTC Quality of Life Questionnaire for Stomach Cancer (EORTC QLQ-STO22). Among the 45 patients available at the one-year follow up after radiation therapy, 40 patients completed the EORTC QLQ-STO22 questionnaire. Their median age was 54.5 (range, 20-70 years). Social functioning was most adversely affected among the respondents with a score of 59, whereas other functions and the global scales were preserved above a score of 70 by linearly transformed values. Fatigue, the financial impact and specific emotional problems such as "thinking about their illness" (STO-ANX) and "worry about weight loss or future health" (STO-BI) were persistently bothersome for some patients. Other stomach-related symptoms such as dysphagia, pain, or reflux were negligible at 1 year after treatment. Therefore, this organ-preserving combined approach was effective for the maintenance of the QOL and minimization of stomach abnormalities in patients with gastric lymphoma. PMID- 18431553 TI - Study of the effects of flexion on the position of the conus medullaris. AB - BACKGROUND: Previous anatomic studies have shown the conus medullaris to terminate between T12 and L1 vertebral levels in adults with normal spinal anatomy. Prior anatomic and radiographic studies of conus position with flexion and extension of the spine have had conflicting results. We performed a cadaveric study with direct visualization of the conus during flexion and extension to further study this question and potentially determine if flexion and extension of the spine during magnetic resonance imaging may prove to be a diagnostic tool in such pathologies as occult tethered cord syndrome. MATERIALS AND METHODS: We performed T12 to L3 laminectomies in five fresh adult cadavers and exposed the conus and cauda equina. Cadavers were fully flexed at the waist and neck and the conus position observed. RESULTS: In all specimens, the conus terminated between T12 and L1 in the prone position. During flexion, the conus position remained stable with no ascent. However, flexion produced stretching and medial displacement of the cauda equina. CONCLUSIONS: Flexion of the spine does not cause the conus medullaris to change position in fresh human cadavers; however, flexion does cause the cauda equina to stretch and displace medially over the conus. Therefore, it is unlikely that the conus would change position during spine flexion during imaging or procedures such as lumbar puncture. PMID- 18431550 TI - Graft failure: III. Glaucoma escalation after penetrating keratoplasty. AB - Glaucoma after penetrating keratoplasty is a frequently observed post-operative complication and is a risk factor for graft failure. Penetrating keratoplasty performed for aphakic and pseudophakic bullous keratopathy and inflammatory conditions are more likely to cause postoperative glaucoma compared with keratoconus and Fuchs' endothelial dystrophy. The intraocular pressure elevation may occur immediately after surgery or in the early to late postoperative period. Early postoperative causes of glaucoma include pre-existing glaucoma, retained viscoelastic, hyphema, inflammation, pupillary block, aqueous misdirection, or suprachoroidal hemorrhage. Late causes include pre-existing glaucoma, angle closure glaucoma, ghost cell glaucoma, suprachoroidal hemorrhage, and steroid induced glaucoma. Determining the cause of IOP elevation can help guide therapeutic intervention. Treatments for refractory glaucoma include topical anti glaucoma medications such as beta-adrenergic blockers. Topical carbonic anhydrase inhibitors, miotic agents, adrenergic agonists, and prostaglandin analogs should be used with caution in the post-keratoplasty patient, because of the possibility of corneal decompensation, cystoid macular edema, or persistent inflammation. Various glaucoma surgical treatments have reported success in post-keratoplasty glaucoma. Trabeculectomy with mitomycin C can be successful in controlling IOP without the corneal toxicity noted with 5-fluorouracil. Glaucoma drainage devices have successfully controlled intraocular pressure in postkeratoplasty glaucoma; this is, however, associated with increased risk of graft failure. Placement of the tube through the pars plana may improve graft success compared with implantation within the anterior chamber. In addition, cyclophotocoagulation remains a useful procedure for eyes that have refractory glaucoma despite multiple surgical interventions. PMID- 18431554 TI - Intradiploic cavernous hemangioma of the skull in a child: a case report. AB - INTRODUCTION: Intradiploic cavernous hemangioma of the skull is seen rarely. Intradiploic cavernous hemangiomas arise from the intrinsic vasculature of the bone and generally picks up at the fourth and fifth decades. DISCUSSION AND CONCLUSION: In this report, we present a 16-year-old child who was admitted with a swelling lesion in the right parietal bone and diagnosed as cavernous hemangioma after total extirpation. PMID- 18431555 TI - Adolf Wallenberg (1862-1949): physician and neuroanatomist. PMID- 18431556 TI - [Molecular pathologic KRAS mutation analysis. A prerequisite of effective antibody treatment for metastasized colorectal cancer]. PMID- 18431557 TI - [Results from a survey of intensive care training. How much that is learned applies to later practice?]. AB - BACKGROUND: There is no doubt that good knowledge of intensive care medicine is necessary for any surgeon, since major surgery entails postoperative intensive care requiring the surgeon's involvement. Recent changes in the German surgical training program, demands for the board examination, and further specialisation have raised the question whether the time spent in ICU education is still adequate and covers topics for young surgeons attendant to their personal professional aims. The present survey was performed to elucidate this topic. METHODS: Questionnaires were sent to 300 randomly chosen surgical residents. RESULTS: Of the questionnaires, 44% were returned and eligible for analysis: 95% considered their ICU education important, 32% worked longer than the (required) 6 months on the ICU, and 62% thought the time spent on ICU was adequate, whereas 14.5% thought it too long. Most of these fellows (84%) worked in university hospitals of large medical centers. After their ICU rotation, the majority felt familiar with the basic procedures and regarded them important also for future work. More advanced techniques (e.g. specifics of artificial ventilation or invasive haemodynamic monitoring) were considered less relevant or not at all. CONCLUSION: The majority of fellows (95%) considered ICU training important, but 50% felt that advanced ICU techniques were not relevant to their planned future work as surgeons. Especially in university hospitals and large medical centers, there were discrepancies between proposed and actually served ICU time as well as between the training program and the methods and techniques the young surgeons felt important for the future. This information may be useful when discussing requirements of surgical education programs. PMID- 18431558 TI - [Cardiovascular events during World Cup Soccer 2006]. PMID- 18431559 TI - [A 68-year-old patient with atrial flutter/fibrillation, inadequate anticoagulation, subacute amaurosis, normal ESR, and lymphadenopathy]. AB - We present a rare manifestation of chronic lymphatic leukemia with progressive bilateral visual loss and the typical fundoscopic picture of anterior ischemic optic nerve neuropathy (AION). Clinical symptoms were due to meningeal metastases and tumor cell infiltration of the optic nerve. The diagnostic clue was provided by lumbar puncture with pressure measurement, which made it possible to differentiate AION from papillitis and papilledema. In this case the patient was able to regain his initial visual activity after intrathecal and systemic polychemotherapy. PMID- 18431560 TI - [Uncertain future of orthopedic research]. PMID- 18431561 TI - Use of polyurethane foam and 3-hydroxy-7,8-benzo-1,2,3,4-tetrahydroquinoline for determination of nitrite by diffuse reflectance spectroscopy and colorimetry. AB - Polyurethane foam (PUF) has been suggested as a solid polymeric reagent for determination of nitrite. The determination is based on the diazotization of end toluidine groups of PUF with nitrite in acidic medium followed by coupling of polymeric diazonium cation with 3-hydroxy-7,8-benzo-1,2,3,4-tetrahydroquinoline. The intensely colored polymeric azodye formed in this reaction can be used as a convenient analytic form for the determination of nitrite by diffuse reflectance spectroscopy (c (min) = 0.7 ng mL(-1)). The possibility of using a desktop scanner, digital camera, and computer data processing for the numerical evaluation of the color intensity of the polymeric azodye has been investigated. A scanner and digital camera can be used for determination of nitrite with the same sensitivity and reproducibility as with diffuse reflectance spectroscopy. The approach developed was applied for determination of nitrite in river water and human exhaled breath condensate. PMID- 18431562 TI - Single-drop analysis of various proteases in a cancer cell lysate using a capillary-assembled microchip. AB - Single-drop analysis of two different real sample solutions (2 microL) while simultaneously monitoring the activity of two sets of ten different proteases on a single microfluidic device is presented. The device, called a capillary assembled microchip (CAs-CHIP), is fabricated by embedding square glass sensing capillaries (reagent-release capillaries, RRC) in the polydimethylsiloxane (PDMS) lattice microchannel, and used for that purpose. First, the performance reliability was evaluated by measuring the fluorescence response of twenty caspase-3-sensing capillaries on a single CAs-CHIP, and a relative standard deviation of 1.5-8.2 (% RSD, n = 5 or 10) was obtained. This suggests that precise multiplexed protease-activity sensing is possible by using a single CAs CHIP with multiple RRCs embedded. Then, using a single CAs-CHIP, real sample analysis of the activity of ten different caspases/proteases in cervical cancer (HeLa) cell lysate treated and untreated with the cell-death-inducer drug, doxorubicin, was simultaneously carried out, and a significant difference in enzyme activity between these two samples was observed. These results suggested the usefulness of the CAs-CHIP in the field of drug discovery. PMID- 18431564 TI - Selectivity of voluntary finger flexion during ischemic nerve block of the hand. AB - During ischemic nerve block of an extremity the cortical representations of muscles proximal to the block are known to expand, increasing the overlap of different muscle representations. Such reorganization mimics that seen in actual amputees. We investigated whether such changes degrade voluntary control of muscles proximal to the block. Nine subjects produced brief, isometric flexion force selectively with each fingertip before, during, and after ischemic block at the wrist. We recorded the isometric force exerted at the distal phalanx of each digit, along with electromyographic (EMG) activity from intrinsic and extrinsic finger muscles. Despite paralysis of the intrinsic hand muscles, and associated decrements in the flexion forces exerted by the thumb, index, and little fingers, the selectivity of voluntary finger flexion forces and of EMG activity in the extrinsic finger muscles that generated these forces remained unchanged. Our observations indicate that during ischemic nerve block reorganization does not eliminate or degrade motor representations of the temporarily deafferented and paralyzed fingers. PMID- 18431565 TI - Interleukin-1 beta of Red nucleus involved in the development of allodynia in spared nerve injury rats. AB - Previous studies have indicated that interleukin-1 beta (IL-1 beta) is involved not only in immune modulation, but also in the modulation of pain in both the peripheral and central nervous systems. The current study investigated the expression of IL-1 beta in the brain of rats with spared nerve injury (SNI), using immunohistochemical technique. The results showed that immunoreactive-like IL-1 beta protein was significantly elevated in the Red nucleus (RN) 2 weeks after SNI. To further study the function of IL-1 beta in RN, different doses of IL-1 beta neutralizing antibody (10, 1.0 and 0.1 ng) were microinjected into the RN contralateral to the nerve injury side of neuropathic rats. The results indicated that the higher doses of anti-IL-1 beta antibody (10 and 1.0 ng) significantly attenuated the mechanical allodynia of neuropathic rats. However, administration of 0.1 ng anti-IL-1 beta antibody did not show anti-allodynia effect. These results suggest that IL-1 beta of RN is involved in the development of neuropathic pain in SNI rats. PMID- 18431566 TI - Genomic location and characterisation of nonclassical MHC class I genes in cattle. AB - The cattle major histocompatibility complex (MHC) region contains a variable number of classical class I genes encoding polymorphic, ubiquitously expressed molecules with a role in antigen presentation. Class I cDNA sequences have previously been reported that are thought to derive from putative nonclassical class I genes. We have located four nonclassical class I genes within the cattle genome; three are close to the MIC genes, and one is close to the classical class I genes. The genomic position relative to anchor genes is very similar to the arrangement reported in the pig MHC region. We have designed gene-specific oligonucleotide primers with which to investigate the presence of these genes in distinct and well-defined MHC haplotypes and to assess transcription in different cell types. Analysis and comparison of all sequences allows an assessment of allelic variation in each case. Partial characterisation gives an indication of the possible role and likely importance of each of these genes. PMID- 18431568 TI - A new and efficient phosphate starvation inducible expression system for Lactococcus lactis. AB - A new expression system for Lactococcus lactis was developed. The system is based on a phosphate starvation inducible pstF promoter of L. lactis MG1363. Intracellular beta-galactosidase and secreted alpha-amylase were produced using this tightly regulated system. No evidence of regulatory sites in regions of the 5'-end of the pstF coding sequence was found. High expression levels of the beta galactosidase gene were obtained using the original pstF RBS in a phosphate depleted medium. The results suggested that with the phosphate starvation inducible system, it is possible to achieve expression levels comparable to the ones obtained with the widely used nisin-controlled gene expression system (NICE). A specific beta-galactosidase activity of 670 microkat g(-1) using a phosphate-depleted medium and an alpha-amylase activity of 3.6 microkat l(-1) in a bioreactor cultivation were produced. The advantages of the current expression system include that no prior removal of phosphate from the medium in bioreactor scale is required, and no additions of inducing agents are needed. Furthermore, the system can be operated in L. lactis without introduction of regulatory genes into the host. PMID- 18431567 TI - Support for the minimal essential MHC hypothesis: a parrot with a single, highly polymorphic MHC class II B gene. AB - We characterized the MHC class II B gene in the green-rumped parrotlet, Forpus passerinus. Three approaches were used: polymerase chain reaction amplification using primers complementary to conserved regions of exon 2, sequencing clones from a genomic library, and amplification of exon 2 using species-specific primers. All three methods indicate that there is only a single class II B locus in this species and no pseudogenes. We suggest that this is the ancestral state for birds. The gene is highly polymorphic; 33 alleles were found in a sample of 25 individuals. Variation in exon 2 is concentrated in the peptide binding residues which show a significant excess of non-synonymous substitutions consistent with the operation of selection in maintaining this extraordinary polymorphism. Genomic clones show that major histocompatibility complex (MHC) gene organization is different from that of chickens; the class II A locus is close to II B. These data provide support for the hypothesis that the bird MHC constitutes a "minimal essential MHC" for responding to infectious disease. PMID- 18431569 TI - A conserved Hpa2 protein has lytic activity against the bacterial cell wall in phytopathogenic Xanthomonas oryzae. AB - The type III secretion system (TTSS) proteins form a needle-like structure injecting effector proteins into eukaryotic target cells. Although the TTSS forms an important pathway for bacterium-host interaction, its assembly process in vivo is poorly understood. The process is thought to include the opening of a pore before TTSS proteins are inserted into the bacterial cell wall. The proteins that break the bacterial cell wall have not yet been identified. We hypothesize that a hypersensitive response and pathogenicity (hrp) gene functions to digest the bacterial cell wall because it contains a conserved protein sequence similar to lytic transglycosylase. In this study, we cloned hrp-associated 2 (hpa2) genes from the bacteria Xanthomonas oryzae pathovars. We show in vitro that expressed Hpa2 protein has a lytic activity against bacterial cell walls. The analysis of a loss-of-function mutant of the hpa2 gene suggests that the hpa2 affects bacterial proliferation in host plants and a hypersensitive response in nonhost plants. As this is the first of such enzyme activity identified in the Hrp protein family, we speculate that the Hpa2 contributes to the assembly of the TTSS by enlarging gaps in the peptidoglycan meshwork of bacterial cell walls. PMID- 18431570 TI - Metastatic bone disease: the requirement for improvement in a multidisciplinary approach. AB - The purpose of this study was to assess the referral system, clinical notes and radiographs of patients presenting with metastatic disease of long bones. The study demonstrated that 93% of oncologists did not use a reliable scoring system to assess risk of pathological fracture, and 60% felt an improvement in communication was required. Notes and radiographs were reviewed for 37 patients presenting with femoral metastatic lesions. Sixteen patients had a Mirels' score of greater than 8. Four patients were referred for an orthopaedic opinion. Twelve patients with a score of greater than 8 were not referred; seven of these patients suffered a pathological fracture. Sixteen patients had a Mirels' score of less than 8; none of these patients were referred for an orthopaedic opinion. No pathological fractures occurred. In conclusion, the majority of patients who score above 8 in the Mirels' scoring system are at risk of fracture and do require prophylactic surgery. In keeping with the British Orthopaedic Association (BOA) guidelines, "Metastatic Bone Disease: A Guide to Good Practice", we would recommend a multidisciplinary approach and the use of a recognised scoring system. PMID- 18431571 TI - Altered cyclophosphamide and thiotepa pharmacokinetics in a patient with moderate renal insufficiency. AB - PURPOSE: We report a patient with renal insufficiency (creatinine clearance, CL(cr) = 38 mL/min) who received high-dose chemotherapy with cyclophosphamide (1,500 mg/m(2) day(-1)), thiotepa (120 mg/m(2) day(-1)) and carboplatin (AUC = 5 mg min/mL day(-1)) for four consecutive days. METHODS: Blood samples were collected on day 1 and 3 and plasma levels of cyclophosphamide, its active metabolite 4-hydroxycyclophosphamide, thiotepa, its main metabolite tepa and carboplatin were determined. RESULTS: Pharmacokinetic analyses indicated that the elimination of cyclophosphamide, thiotepa, carboplatin, but especially tepa was strongly reduced in this patient, resulting in increased exposures to these compounds of 67, 43, 30 and 157%, respectively, compared to a reference population (n = 24) receiving similar doses. Exposure to 4 hydroxycyclophosphamide increased 11%. CONCLUSION: These results suggest that it may not be necessary to alter the dose of cyclophosphamide in patients with moderate renal impairment. However, because high exposures to thiotepa and tepa have been correlated with increased toxicity, caution should be applied when administering thiotepa to patients with renal insufficiency. PMID- 18431572 TI - Delineation of the interactions between the chemotherapeutic agent eribulin mesylate (E7389) and human CYP3A4. AB - PURPOSE: Eribulin mesylate (E7389), a structurally simplified, synthetic analog of the marine natural product halichondrin B, acts by inhibiting microtubule dynamics via mechanisms distinct from those of other tubulin-targeted agents. Eribulin is currently in Phase III clinical trials for the treatment of metastatic breast cancer. Since drug-induced modulation of cytochrome P450 enzymes, particularly CYP3A4, is a frequent cause of drug-drug interactions, we examined the effects of eribulin on the activity and expression of hepatic and recombinant CYP3A4 (rCYP3A4) in vitro. METHODS: Identification of the enzyme(s) responsible for eribulin metabolism was based on compound depletion and metabolite formation in reaction mixtures containing subcellular liver fractions or primary human hepatocytes, plus recombinant Phases I and II metabolic enzymes. The role of the enzyme(s) identified was confirmed using enzyme-selective inhibitors and the correlation with prototypic enzyme activity. The effect of eribulin on enzymatic activity was characterized using both microsomal preparations and recombinant enzymes, while the possible modulation of protein expression was evaluated in primary cultures of human hepatocytes. RESULTS: Eribulin was primarily metabolized by CYP3A4, resulting in the formation of at least four monooxygenated metabolites. In human liver microsomal preparations, eribulin suppressed the activities of CYP3A4-mediated testosterone and midazolam hydroxylation with an apparent K (i) of approximately 20 microM. Eribulin competitively inhibited the testosterone 6beta-hydroxylation, nifedipine dehydration, and R-warfarin 10-hydroxylation activities of rCYP3A4, with an average apparent K (i) of approximately 10 microM. These inhibitions were reversible, with no apparent mechanism-based inactivation. Eribulin did not induce the expression or activities of CYP1A and CYP3A enzymes in human primary hepatocytes, and clinically relevant concentrations of eribulin did not inhibit CYP3A4-mediated metabolism of various therapeutic agents, including carbamazepine, diazepam, paclitaxel, midazolam, tamoxifen, or terfenadine. CONCLUSIONS: Eribulin was predominantly metabolized by CYP3A4. Although eribulin competitively inhibited the testosterone 6beta-hydroxylation, nifedipine dehydration, and R-warfarin 10-hydroxylation activities of rCYP3A4, it did not induce or inhibit hepatic CYP3A4 activity at clinically relevant concentrations. As eribulin does not appear to affect the metabolism of other therapeutic agents by CYP3A4, our data suggest that eribulin would not be expected to inhibit the metabolism of concurrently administered drugs that are metabolized by CYP3A4, suggesting a minimal risk of drug-drug interactions in the clinical setting. PMID- 18431573 TI - Autoantibodies to alfa-fodrin in patients with Hashimoto thyroiditis and Sjogren's syndrome: possible markers for a common secretory disorder. AB - Presence of autoantibodies to alfa-fodrin was investigated in patients with Sjogren's syndrome (n = 61), Hashimoto thyroiditis (n = 27), Sjogren's syndrome associated with Hashimoto thyroiditis (n = 31) and in healthy persons (n = 77). In each group, level of alfa-fodrin antibodies was higher than in the controls. There was no significant difference in their presence either between patients with Hashimoto thyroiditis with or without Sjogren's syndrome, or-in IgA isotype between Sjogren's and Hashimoto thyroiditis patients. Correlation was found between the level of IgG alfa-fodrin and anti-thyroglobulin antibodies. Based on these findings, fodrin can be associated with both endocrine and exocrine glandular secretion. Antibodies to alfa-fodrin might have a role in the pathogenesis of Hashimoto thyroiditis concerning the "final common effectory pathway", secretion. Alfa-fodrin antibodies can be good markers of secretory disorders. Assessment of these autoantibodies might help the diagnosis and follow up of patients with impaired secretory capability of not only autoimmune origin. PMID- 18431574 TI - Clinical impact of the use of additional ultrasonography in diagnostic breast imaging. AB - The degree of adherence with evidence-based guidelines for the use of breast ultrasonography was determined in clinical practice of radiologists in six hospitals. Additional ultrasonography was performed in 2,272 (53%) of all 4,257 patients referred for mammography. High adherence rates (mean: 95%) were observed for guidelines recommending ultrasonography in patients referred for palpable breast masses and abnormal screening and diagnostic mammograms. Lower adherence rates (mean: 81%, Pearson correlation coefficient= -0.57; p=0.001) were found for guidelines advising against additional ultrasonography in patients referred for breast symptoms, a known benign abnormality, a family history or anxiety of breast cancer. The overuse of ultrasonography in 442 patients and underuse in 95 patients led to five additional false-positive results. It was concluded that the guidelines seem workable and feasible in clinical practice and that the current daily routine of diagnostic breast imaging corresponded to a great extent to the guidelines proposed. PMID- 18431575 TI - Total-body contrast-enhanced MRA on a short, wide-bore 1.5-T system: intra individual comparison of Gd-BOPTA and Gd-DOTA. AB - Total-body contrast-enhanced MRA (CE-MRA) provides information of the entire vascular system according to a one-stop-shop approach. Short, wide-bore scanners have not yet been used for total-body CE-MRA, probably due to their restricted field of view in the z-direction. The purpose of this feasibility study is to introduce an image protocol for total-body MRA on a short, wide-bore system. The protocol includes five to six table-moving steps and two injection runs. Two pharmacologically different contrast materials (CM) were applied in ten healthy volunteers in view of possible CM-dependent influences on the protocol outcome (Gd-Bopta, Gd-Dota). Differences consisted of significantly higher CNR with Gd Bopta with a mean of 73.8+/-38.7 versus 69.1+/-34.3 (p=0.008), significantly better arterial visualization values with Gd-Dota with a mean of 1.26+/-0.44 versus 1.53+/-0.73 (p=0.003) and a tendency to less venous overlay with Gd-Dota, mean 1.19+/-0.44 and 1.34+/-0.72, respectively (p=0.065) (two-tailed Wilcoxon matched-pairs test). Overall 94% of the steps were valued as qualitatively excellent or good. The good results with both CM suggest a transfer to further patient evaluation. PMID- 18431576 TI - Perfusion CT measurements in healthy cervical spinal cord: feasibility and repeatability of the study as well as interchangeability of the perfusion estimates using two commercially available software packages. AB - Our purpose was to examine the feasibility and reproducibility of perfusion CT studies in the cervical spinal cord and the interchangeability of the values obtained by two post-processing methods. The perfusion CT studies of 40 patients with neck tumours were post-processed using two software packages (Software-1: deconvolution-based analysis with adiabatic tissue homogeneity approach and Software-2: maximum-slope-model with Patlak analysis). Eight patients were examined twice for assessing the reproducibility of the technique. Two neuroradiologists separately post-processed the images with two arterial input functions (AIFs): (1) the internal carotid artery (ICA) and (2) the vertebral artery (VA). Maps of blood flow (F) in ml/min/100 g, blood volume (V) in ml/100 g, mean transit time (MTT) in seconds (s) and permeability (PS) in ml/min/100 g were generated. The mean F, V, MTT and PS (Software-1) with VA-AIF and ICA-AIF were 8.93, 1.12, 16.3, 1.88 and 8.57, 1.19, 16.85 and 1.94, respectively. The reproducibility of the techniques was satisfactory, while the V and MTT values (in Software-1) and the F and V values (in Software-2) were dependent on the site of the AIF (p >or= 0.03 and p=0.02, respectively). The interobserver agreement was very good. The significant differences in measurements for a single patient (%) using Software-1/Software-2 were +/-120%/110%, 90%/80%, 180% and 250%/130% for F, V, MTT and PS, respectively. Only F and PS values in the healthy tissue seemed to be interchangeable. Our results were in essential agreement with those derived by invasive measurements in animals. The cervical spine perfusion CT studies are feasible and reproducible. The present knowledge has to be validated with studies in spinal cord tumours in order to decide the usefulness of the perfusion CT in this field. PMID- 18431577 TI - Digital chest radiography: an update on modern technology, dose containment and control of image quality. AB - The introduction of digital radiography not only has revolutionized communication between radiologists and clinicians, but also has improved image quality and allowed for further reduction of patient exposure. However, digital radiography also poses risks, such as unnoticed increases in patient dose and suboptimum image processing that may lead to suppression of diagnostic information. Advanced processing techniques, such as temporal subtraction, dual-energy subtraction and computer-aided detection (CAD) will play an increasing role in the future and are all targeted to decrease the influence of distracting anatomic background structures and to ease the detection of focal and subtle lesions. This review summarizes the most recent technical developments with regard to new detector techniques, options for dose reduction and optimized image processing. It explains the meaning of the exposure indicator or the dose reference level as tools for the radiologist to control the dose. It also provides an overview over the multitude of studies conducted in recent years to evaluate the options of these new developments to realize the principle of ALARA. The focus of the review is hereby on adult applications, the relationship between dose and image quality and the differences between the various detector systems. PMID- 18431578 TI - Intrinsic respiratory gating in small-animal CT. AB - Gating in small-animal CT imaging can compensate artefacts caused by physiological motion during scanning. However, all published gating approaches for small animals rely on additional hardware to derive the gating signals. In contrast, in this study a novel method of intrinsic respiratory gating of rodents was developed and tested for mice (n=5), rats (n=5) and rabbits (n=2) in a flat panel cone-beam CT system. In a consensus read image quality was compared with that of non-gated and retrospective extrinsically gated scans performed using a pneumatic cushion. In comparison to non-gated images, image quality improved significantly using intrinsic and extrinsic gating. Delineation of diaphragm and lung structure improved in all animals. Image quality of intrinsically gated CT was judged to be equivalent to extrinsically gated ones. Additionally 4D datasets were calculated using both gating methods. Values for expiratory, inspiratory and tidal lung volumes determined with the two gating methods were comparable and correlated well with values known from the literature. We could show that intrinsic respiratory gating in rodents makes additional gating hardware and preparatory efforts superfluous. This method improves image quality and allows derivation of functional data. Therefore it bears the potential to find wide applications in small-animal CT imaging. PMID- 18431579 TI - Retroperitoneal nephroureterectomy with excision of cuff of the bladder for upper urinary tract transitional cell carcinoma: comparison of laparoscopic and open surgery with long-term follow-up. AB - OBJECTIVES: To compare outcome and long term follow up of laparoscopic retroperitoneal nephroureterectomy with excision of cuff of bladder (LRPNUT-BCE) and standard open nephroureterectomy with excision of cuff of bladder (ONUT-BCE) in patients of upper urinary tract transitional cell carcinoma (UTTCC). MATERIAL AND METHODS: From January 1998 to October 2006, we have performed over 55 cases of nephroureterectomy with excision of cuff of bladder for UTTCC. Out of these, 48 patients undergoing LRPNUT-BCE and ONUT-BCE were categorized retrospectively into group A (21), and group B (27), respectively. The clinical data including intraoperative, postoperative, and follow-up data was recorded for two groups and analyzed statistically. RESULTS: The mean operating time was significantly higher in-group A (P < 0.001). The different techniques of bladder cuff excision were used in group A. The mean analgesic use, hospital stay and weeks to normal activity were significantly lesser in-group A (P < 0.001). The intraoperative, postoperative complications and mean follow up were comparable in two groups. There was no local recurrence in either group. The bladder recurrence and distant metastases were comparable in two groups (group A--9.52%, group B--11.1% and group A--9.52%, group B--11.1%, respectively). The 5-year recurrence free, cancer specific and overall survivals were comparable in two groups. CONCLUSIONS: On long-term follow-up of oncological efficacy, LRPNUT-BCE satisfactorily compares with traditional open surgery (ONUT-BCE) with longer operating time, while decreasing postoperative morbidity and extending benefits of minimally invasive surgery to the patients. PMID- 18431580 TI - [Bilateral chorioretinitis with hilus lymphoma. Differential diagnosis and treatment]. PMID- 18431581 TI - [Fusarium keratitis]. AB - The case of a previously healthy 48-year-old patient (not a contact lens wearer) with a Fusarium keratitis is reported. He had developed a deep corneal ulcer with total stromal infiltration (Fusarium ssp.). An intensive topical and systemic antiinfectious medication was initiated, along with some procedures (keratoplasty, rinsing of the anterior chamber). However, the infection progressed (endophthalmitis), and an enucleation was necessary. Despite modern drug therapy and surgical intervention, fungal keratitis can cause an enucleation. PMID- 18431582 TI - [A prospective intraindividual comparison between laser in situ keratomileusis and laser subepithelial keratectomy for myopia. 1-year follow-up results]. AB - BACKGROUND: The aim of this study was to intraindividually compare the outcome of laser in situ keratomileusis (LASIK) and laser subepithelial keratectomy (LASEK) in patients with similar refraction in both eyes when treated with two different procedures. PATIENTS AND METHODS: In this prospective nonrandomized study, 30 myopic patients with and without astigmatism were treated by Lasik in one eye and LasEk in the fellow eye. All eyes were treated using the MEL 70-G Excimer laser (Carl Zeiss Meditec, Germany). Uncorrected visual acuity (UCVA) and best spectacle-corrected visual acuity (BSCVA) as well as photopic contrast sensitivity (Pelli-Robson charts) and scotopic contrast sensitivity (Nyktometer, Rodenstock, Germany) were evaluated. The patients were also asked about their preferences regarding the procedures. RESULTS: The preoperative spherical equivalent (SE) was -4.54+/-1.64 D and the mean BSCVA was 0.94 in Lasik eyes. The SE was -3.9+/-1.57 D and the BSCVA was 0.89 in LASEK eyes. The mean 1-year post operative SE was -0.33+/-0.48 D in the Lasik eyes and -0.19+/-0.26 D in the LASEK eyes. Efficacy and safety were similar in the two groups, but Lasik eyes had a greater standard deviation at 1 year. Pelli-Robson contrast sensitivity was 10.75+/-3.05 in the Lasik eyes compared with 11.4+/-3.21 in the fellow LASEK eyes. Seventeen patients named Lasik and 12 patients named LASEK as their procedure of choice. CONCLUSION: Both procedures enable similarly good results. LASEK showed a tendency to a smaller standard deviation in final refraction and better contrast sensitivity. PMID- 18431583 TI - [Longitudinal effects of structured patient education programs for vasculitis patients]. AB - According to the literature it is known that structured standardized patient education represents an effective additional treatment in patients with chronic diseases. Positive effects in the reduction of disease activity and depression have been shown for rheumatoid arthritis, systemic lupus erythematodes, and diabetes mellitus. An interdisciplinary approach for providing information was developed for patients with primary systemic vasculitides (PSV) in the vasculitis center in Bad Bramstedt. The contents of the seminars were revised and condensed into five modules. To evaluate the new form of the program a documentation system was designed. Patients were trained in closed groups (n=10-15) and completed the questionnaires at baseline, 4 weeks, 6 and 12 months following participation. A total of 102 patients in 10 closed groups showed a statistically significant increase in their knowledge in the three aspects of medicine, therapy and side effects, nutrition and physiotherapy. Health-related quality of life in all dimensions increased considerably. Both self-efficacy and the patient-assessed health status improved. The standardized structured education program for vasculitis patients provides an additional treatment in the interdisciplinary care of vasculitis. PMID- 18431584 TI - [A view Down Under. Self-management initiatives and patient education in Australia]. AB - The growing burden of chronic disease and the current nature of healthcare systems which are structurally ill-equipped to cater to the complex needs of patients with chronic conditions has led to governments and healthcare providers seeking alternative ways to improve patients own capacity to actively self-manage their chronic condition. In Australia, there has been a focus on patient education and self-management programs within the healthcare system to achieve this. These programs aim to empower patients through providing information and teaching skills and techniques to improve self-care and doctor-patient interaction with the ultimate goal of improving quality of life. Patient education and self-management programs have been supported through several national government initiatives and implemented within the healthcare setting. This paper describes the current position of patient education and self management within the Australian healthcare system. It further describes a new collaboration project between an Australian and a German research team which aims at translating an assessment questionnaire used in Australia for the evaluation of self-management programs, the "Health Education Impact Questionnaire" ("heiQ"); this instrument is expected to be of significant use in the German rehabilitative system. PMID- 18431585 TI - [Patient education: cui bono?]. PMID- 18431586 TI - Laparoscopy-assisted cystectomy for large adnexal cysts. AB - OBJECTIVE: To evaluate the feasibility and surgical outcome of laparoscopy assisted surgery for large adnexal cysts. METHODS: From January 1998 to October 2007, 46 women underwent laparoscopy-assisted surgery for large adnexal cysts whose maximum diameter were between 10 and 20 cm, radiologic and laboratory features suggestive of benign disease. All the patients had a pre-operative ultrasound with or without computed tomography and CA-125 assessment. Patients' demographics, clinical and ultrasound features, CA-125 values, surgical procedures, operative and post-operative complications, estimated amount of blood loss (EBL), operative time, conversion to laparotomy and the pathological findings were recorded. RESULTS: Fourty-six consecutive patients underwent laparoscopy-assisted surgery over 9 years. The mean and range of the patients' age and body mass index were 34.1 +/- 6.3 and (21-45) years and 27.4 +/- 5.9 and (22-40), respectively. In all the patients, except one with borderline ovarian tumor, laparoscopy-assisted surgery was successful. There were no operative or post-operative complications. The mean and range of the operative time, EBL and hospital stay were 48.4 +/- 7.3 and (35-65) min, 55.0 +/- 28.9 and (25-150) mL, 1.49 +/- 0.50 and (1-3) days, respectively. The mean and range of the extracorporeal cystectomy time were 10.2 +/- 2.7 and (8-14) min. The surgical procedures performed were: ovarian and paraovarian cystectomy (n = 45), unilateral salpingo-oophorectomy, pelvic-paraaortic lymphadenectomy and omentectomy (n = 1). Pathologic findings included serous cystadenoma (n = 26), mucinous cystadenoma (n = 7), dermoid (n = 6), endometriosis (n = 6), and borderline ovarian tumor (n = 1). CONCLUSION: Laparoscopy-assisted surgery is feasible and safe for women with large benign adnexal cysts and result s in a short surgery time. PMID- 18431587 TI - Combined pars plana vitrectomy and artificial iris diaphragm implant after globe rupture. AB - BACKGROUND: Retinal detachment (RD) associated with aniridia due to globe rupture (GR) is an uncommon condition with a severe prognosis. Surgical technique must address anterior and posterior segment issues secondary to the altered compartmentalization and increased risk for corneal toxicity. The purpose of this paper is to report a series of GR patients undergoing combined pars plana vitrectomy (PPV) and artificial iris diaphragm (AID) implant for the repair of RD associated to aniridia. METHODS: The authors retrospectively reviewed 12 consecutive patients operated on by a single surgeon. Surgery consisted of a standard three-port PPV with extensive bimanual dissection of vitreous base and ciliary body membrane and combined AID implant. Office visits included Snellen visual acuity (VA), intraocular pressure measurement, biomicroscopy and indirect ophthalmoscopy. AID prosthesis included aniridic IOLs, Heimann's PMMA and silicone diaphragm. RESULTS: Mean age was 53 years and mean follow-up was 19 months. At the end of follow-up, seven patients gained more than two lines (58.3%), two lost their vision (16.6%) and three were unchanged (25%). Seven patients (58.3%) had a VA better than 20/400 and one (8%) 20/40 vision. Eight patients (66.6%) retained a clear cornea, two (16.6%) had minimal corneal oedema and two (16.6%) corneal decompensation. Implanted prosthesis included two silicone diaphragms, four PMMA diaphragms and six aniridic IOLs. After an average 1.6 operations, the retina was completely attached in six patients (50%), partially attached in four (33.3%) and detached in two (16.6%). CONCLUSION: RD associated to GR carries a guarded prognosis both due to RD complexity and hypotony. The combined repair of RD and aniridia after GR offers the advantage of addressing all issues at one time allowing correct eye compartmentalization and better tamponade effect. Successful anatomical and functional results can be achieved although multiple surgeries are often needed. PMID- 18431588 TI - One-year results of a multicenter controlled clinical trial of triamcinolone in pars plana vitrectomy. AB - PURPOSE: To evaluate the 1-year results of using triamcinolone acetonide (TA) in pars plana vitrectomy (PPV). DESIGN: Multicenter prospective controlled clinical trial. METHODS: SETTING AND STUDY POPULATION: the study population comprised 774 eyes from patients treated at eight Japanese hospitals, among which 391 eyes underwent TA-assisted PPV and 383 control eyes underwent conventional PPV. The patients were assigned to the two groups using a single-blind quasi-randomization approach within the participating clinical centers. INTERVENTION: intra-operative use of TA to aid visualization of the vitreous. MAIN OUTCOME MEASURES: changes of visual acuity, post-operative complications (including additional surgery), and adverse events occurring within 1 year of the operation were compared between the TA-PPV group and the conventional PPV group. RESULTS: The visual acuity improved over time, and no significant differences were found between the two groups (log rank versus TA, P = 0.98 for improvement, P = 0.26 for deterioration). The logistic regression model also showed that the intra-operative use of TA was not a significant factor for the improvement of visual acuity [P = 0.91, odds ratio (OR) = 1.10, 95% confidence interval (95%CI) = 0.860-1.183)] after adjustments for age, gender, and diagnosis. Intra-operative TA was not a significant factor for the need for additional surgery (log-rank test P = 0.45, logistic regression test P = 0.35, OR = 1.23, 95%CI = 0.797-1.911]. No serious adverse events related to surgery were observed. CONCLUSIONS: This 1-year follow-up study of a controlled clinical trial showed that TA-assisted PPV had neither a positive nor a negative effect on visual acuity, the incidence of additional surgeries, or adverse events compared with conventional PPV. PMID- 18431589 TI - High-mobility group box 1 protein in endophthalmitis. AB - BACKGROUND: High-mobility group box 1 protein (HMGB1) is recently described as a late mediator of lethal endotoxemia with proinflammatory cytokine-like properties. The purpose of this study was to determine whether HMGB1 is involved in endophthalmitis. METHODS: In this retrospective case-control study, vitreous levels of HMGB1 were measured by an enzyme-linked immunosorbent assay in ten eyes with endophthalmitis, and in 12 eyes with idiopathic macular holes which served as controls. Formalin-fixed and paraffin-embedded tissue sections of an enucleated eye with endophthalmitis, and a control eye with recurrent conjunctival malignant melanoma, were analyzed by immunohistochemistry with an anti-HMGB1 antibody. RESULTS: The vitreous HMGB1 level of the patients with endophthalmitis was 13.96 +/- 17.17 ng/ml (mean+/-SD), which was significantly higher than that of the controls (0.236 +/- 0.128 ng/ml; P = .0006, Mann-Whitney U test). There were significant correlations between HMGB1 level and disease duration, presenting visual acuity, and final visual acuity. In the eye with endophthalmitis, HMGB1 expression was diffusely observed, particularly in the extranuclear region of the retina and the choroid with infiltrating inflammatory cells. CONCLUSION: HMGB1 can be released in the vitreous of eyes with endophthalmitis depending on inflammation and tissue damage. Our results suggest that HMGB1 may be a late mediator of endophthalmitis, and related to the progression of endophthalmitis. PMID- 18431590 TI - Proliferation of the vascular endothelium of the iris following total debridement of the corneal epithelium and limbal excision of rabbits. AB - BACKGROUND: Damage to the corneal epithelium causes not only a reaction for its repair but also affects other parts of the cornea as well as different components of the anterior segment of the eye. The purpose of this investigation was to analyze the consequences, following epithelial and limbal damage, to the iris of rabbits (Oryctolagus cuniculus). METHODS: The corneal epithelium was thoroughly scraped followed by surgical excision of the limbus. Next, (3)H-thymidine ((3)H TdR) was injected intravitreally both into the right (experimental) and left (control) eyes which had their anterior segments processed for autoradiography at intervals of 2, 7 and 21 days after surgery (three rabbits per interval). The irises were also examined with scanning-electron and confocal microscopy after Evans blue injection. RESULTS: There was a high frequency of labeling in the cells of the iris blood vessels in the experimental eye, particularly the endothelial ones. The ratio of labeled cells between experimental and control irises was 40:1, with a population of nuclei increasing by 25% and remaining labeled up to 21 days. There was also an increase in the volume of the iris vasculature as shown by confocal microscopy. The high labeling frequencies of the vascular cells were observed throughout the iris from the ciliary to the pupillary regions. CONCLUSIONS: The lesions on the corneal epithelium elicit proliferation of the iris vascular cells, mainly its endothelium, as well as an early breakdown of the blood-aqueous barrier. The daughter cells resulting from the damage to the eye surface were detected up to 21 days after a single injection of (3)H-TdR, most likely due to their slow turnover. As a consequence of this proliferation, the vasculature of the iris increased in volume. PMID- 18431591 TI - Feasibility study of chitosan as intravitreous tamponade material. AB - BACKGROUND: Chitosan can inhibit fibroblastic proliferation by suppressing fibroblast cells, and has the similar physiological characteristics as normal vitreous body, so it might have the potential to become vitreous filling material and might possibly inhibit proliferative vitreous retinopathy. To investigate the possibility of chitosan as vitreous filling material, this study was designed to investigate retina, ciliary body, lens and cornea morphology changes, intraocular pressure and intraocular inflammatory factors fluctuating after chitosan intravitreous tamponade. METHODS: Fifteen healthy chinchilla rabbits were chosen; three of them were a blank (negative) control group without any surgical procedure. The remaining 12 rabbits received vitrectomy on both eyes; all the right eyes (experimental group) were given 1.2-1.8 ml (average 1.5 ml) of chitosan intravitreously, while sodium hyaluronate were given in the left eyes (control group). All eyes underwent slit-lamp biomicroscope and indirect ophthalmoscope examination and intraocular pressure measurement pre- and post-op. The concentration of IL-6, IL-8 (radioimmunoassay), NO (nitrate reductase method) in aqueous humor and vitreous body were tested at day 15 and day 30 post-op. At day 30 post-op, the cornea, ciliary body, and lens were dissected for light microscopy examination, and the retinal tissues 2PD away from the optic disc on the vertical orientation of posterior pole were dissected for light- and electro microscope examination. RESULTS: The conjunctival congestion and slight inflammatory response in the anterior chamber disappeared within 7 days post-op. During the 30-day experiment, cornea, lens and the filling material in vitreous cavity were transparent in all animals. The retina was attached without proliferation. The intraocular pressure in the experimental group post-op fluctuated between 4.55 +/- 2.94 and 6.25 +/- 2.37 mmHg, which was not significantly different from the situation pre-op (6.18 +/- 1.19 mmHg) (P > 0.05). The intraocular pressure in the control group post-op fluctuated between 5.10 +/- 2.51 and 5.90 +/- 2.49 mmHg, which was not significantly different from the situation pre-op (6.50 +/- 0.94 mmHg) (P > 0.05). There was also no significant difference in the intraocular pressure post-op at different time points between the experimental group and control group (all P > 0.05). At day 15 post-op, IL-6 concentration in aqueous humor were 37.31 +/- 8.59 ng/ml and 39.52 +/- 9.69 ng/ml in experimental group and control group respectively, both higher than those in the blank control group (26.55 +/- 9.34 ng/ml) (P < 0.05). IL-8 concentration were 7.00 +/- 3.79 ng/ml and 6.32 +/- 3.68 ng/ml respectively, no significant difference to the blank control group (4.72 +/- 1.71 ng/ml) (P > 0.05): the concentrations of NO were 63.94 +/- 26.80 micromol/ml and 51.81 +/- 13.19 micromol/ml respectively, no significant difference to the blank control group (50.36 +/- 15.67 micromol/ml) (P > 0.05). At day 30 post-op, the concentrations of IL-6, IL-8 and NO in aqueous humor showed no significant difference among all three groups (P > 0.05). In vitreous body at day 30 post-op, the concentrations of IL-8 in experimental group and control group were 10.17 +/- 3.63 ng/ml and 10.69 +/- 3.52 ng/ml, and those of NO were 50.23 +/- 19.69 micromol/mL and 50.60 +/- 12.72 micromol/mL respectively, all higher than in the blank control group (30.37 +/- 14.63 micromol/ml) (P < 0.05); the concentrations of IL-6 were 24.51 +/- 10.71 ng/ml and 26.36 +/- 13.00 ng/ml, no significant difference to the blank control group (24.06 +/- 5.98 ng/ml) (P > 0.05). At various time points, there was no significant difference in the concentrations of IL-6, IL-8 and NO in aqueous humor and vitreous body in the experimental group and the control group (P > 0.05). There was no morphological change found under light microscopy in cornea, ciliary body and lens. The outer plexiform layer of retina was thinner, but no significant degeneration, necrosis, karyopyknosis or lysis were found under the ultrastructural microscopy. CONCLUSION: Chitosan intravitreous tamponade has no significant effect on the histology of the eye, doesn't cause intraocular pressure to fluctuate, and slightly increases inflammatory factors (IL-6, IL-8, NO) in comparison to the normal levels, but with no significant difference from the effects caused by sodium hyaluronate, which indicated chitosan might not lead to a clinically significant inflammatory response. It suggests that chitosan could be used as intravitreous tamponade material. PMID- 18431592 TI - Changes in central artery blood pressure and wave reflection during a cold pressor test in young adults. AB - The relative contribution of sympathetic nervous system (SNS)-induced increase in peripheral vascular resistance on central artery blood pressure (BP) and aortic wave reflection (augmentation index; AIx) is not completely understood. Central BP and wave reflection characteristics were measured using radial artery applanation tonometry before, during a 3-min cold pressor test (CPT), and 90 and 180-s post-CPT in 15 young, healthy adults (25 +/- 1 years). The CPT resulted in a greater magnitude of change in the estimated aortic systolic (31 vs. 23%, P < 0.05) and pulse (31 vs. 13%, P < 0.05) BP compared with the change in brachial artery BP. Additionally, the CPT resulted in an increased mean arterial pressure (MAP) (P < 0.05) and AIx (10 +/- 2 vs. 26 +/- 2%, P < 0.05). The change in MAP during the CPT was correlated to the change in AIx (r = 0.73, P < 0.01) and inversely related to roundtrip duration of the reflected wave to the periphery and back (r = -0.57, P < 0.05). The present study suggests that cold pressor testing results in a significant increase in arterial wave reflection intensity, possibly due to an increased MAP. However, the greater increase in systolic and pulse BP in the central compared with the peripheral circulation suggests that increased central artery wave reflection intensity contributes to increased left ventricular myocardial oxygen demand during CPT-induced hypertension. PMID- 18431593 TI - Polytrauma--pathophysiology and management principles. AB - BACKGROUND: Multiple injury results in a complex pathophysiological and immunological response. Depending on the individual injury pattern, the time elapsed after injury, and the systemic "danger response", the surgical treatment has to be modified. OBJECTIVES: This overview provides new insights in the pathophysiology of the early danger response after polytrauma and outlines the main resulting consequences for surgical management. RESULTS: First, synchronically to the clinical assessment, life-saving procedures need to be performed rapidly, such as control of massive intra-thoracic or abdominal bleeding and decompression of the chest and brain, as standardized by advanced trauma life support guidelines. During the second phase of "day-one-surgery" damage-control interventions such as debridement, decompression and temporary fracture stabilization are needed to avoid an excessive molecular and cellular danger response. Trauma-adjusted surgical techniques are crucial to limit the systemic response known to put remote organs at risk. In the "vulnerable phase" when the patient's defense is rather uncontrolled, only "second look" debridement to minimize a "second hit" is recommended. After stabilization of the patient as indicated by improvement of tissue oxygenation, coagulation, and decreased inflammatory mediators, "reconstructive surgery" can be applied. CONCLUSION: Individually adjusted surgical "damage control" and "immune control" are important interactive concepts in polytrauma management. PMID- 18431595 TI - Natural variation of potato allene oxide synthase 2 causes differential levels of jasmonates and pathogen resistance in Arabidopsis. AB - Natural variation of plant pathogen resistance is often quantitative. This type of resistance can be genetically dissected in quantitative resistance loci (QRL). To unravel the molecular basis of QRL in potato (Solanum tuberosum), we employed the model plant Arabidopsis thaliana for functional analysis of natural variants of potato allene oxide synthase 2 (StAOS2). StAOS2 is a candidate gene for QRL on potato chromosome XI against the oomycete Phytophthora infestans causing late blight, and the bacterium Erwinia carotovora ssp. atroseptica causing stem black leg and tuber soft rot, both devastating diseases in potato cultivation. StAOS2 encodes a cytochrome P450 enzyme that is essential for biosynthesis of the defense signaling molecule jasmonic acid. Allele non-specific dsRNAi-mediated silencing of StAOS2 in potato drastically reduced jasmonic acid production and compromised quantitative late blight resistance. Five natural StAOS2 alleles were expressed in the null Arabidopsis aos mutant under control of the Arabidopsis AOS promoter and tested for differential complementation phenotypes. The aos mutant phenotypes evaluated were lack of jasmonates, male sterility and susceptibility to Erwinia carotovora ssp. carotovora. StAOS2 alleles that were associated with increased disease resistance in potato complemented all aos mutant phenotypes better than StAOS2 alleles associated with increased susceptibility. First structure models of 'quantitative resistant' versus 'quantitative susceptible' StAOS2 alleles suggested potential mechanisms for their differential activity. Our results demonstrate how a candidate gene approach in combination with using the homologous Arabidopsis mutant as functional reporter can help to dissect the molecular basis of complex traits in non model crop plants. PMID- 18431596 TI - Brassica napus possesses an expanded set of polygalacturonase inhibitor protein genes that are differentially regulated in response to Sclerotinia sclerotiorum infection, wounding and defense hormone treatment. AB - Most plants encode a limited set of polygalacturonase inhibitor (PGIP) genes that may be involved in aspects of plant development, but more importantly in the inactivation of polygalacturonases (PG) secreted by pathogens. Previously, we characterized two Brassica napus PGIP genes, BnPgip1 and BnPgip2, which were differentially expressed in response to pathogen infection and wounding. Here we report that the B. napus genome encodes a set of at least 16 PGIP genes that are similar to BnPgip1 or BnPgip2. This is the largest Pgip gene family reported to date. Comparison of the BnPGIPs revealed several sites within the xxLxLxx region of leucine rich repeats that form beta-sheets along the interacting face of the PGIP that are hypervariable and represent good candidates for generating PGIP diversity. Characterization of the regulatory regions and RT-PCR studies with gene-specific primers revealed that individual genes were differentially responsive to pathogen infection, mechanical wounding and signaling molecules. Many of the BnPgip genes responded to infection by the necrotic pathogen, Sclerotinia sclerotiorum; however, these genes were also induced either by jasmonic acid, wounding and salicylic acid or some combination thereof. The large number of PGIPs and the differential manner in which they are regulated likely ensures that B. napus can respond to attack from a broad spectrum of pathogens and pests. PMID- 18431597 TI - Establishment of CXCR4-small interfering RNA retrovirus vector driven by human prostate-specific antigen promoter and its biological effects on prostate cancer in vitro and in vivo. AB - PURPOSE: CXC chemokine receptor-4 (CXCR4) is closely involved in bone metastasis of prostate cancer, and CXCR4 levels are frequently increased in prostate cancer cells and tissues. In the present study, its biological effects on prostate cancer in vitro and in vivo and feasibility to be a therapy target were investigated using a RNA interfering retrovirus vector targeting CXCR4 gene driven by human prostate-specific antigen promoter (pPSA). METHODS: We established a pPSA-siCXCR4 retrovirus vector and transfected prostate cancer cell PC-3m, LNCaP and breast cancer cell MCF-7, respectively. The expression of CXCR4 mRNA and protein was detected by RT-PCR and western blot, and the ability of adhesion, migration, invasion of prostate cancer cells was assessed using Transwell chamber. A metastasizing model using BALB/cA mice with human bone tissue implantation was established too, and transfected prostate cancer cells were via caudal vein. Survival time of mice suffering bone metastatic tumor as well as the weight and volume of these tumors were recorded and analyzed. RESULTS: The expression of CXCR4 mRNA and protein in androgen-responsive LNCaP cells was blocked by the pPSA-siCXCR4 vector, but it could not work in non androgen-responsive PC-3m cell and breast cancer cell MCF-7. The results of experiments in vitro also showed that the adhesion, transendothelial migration and invasive ability of transfected LNCaP cells were impaired, while there was no change in PC-3m and MCF-7 cells after transfection. pPSA-siCXCR4 represented a similar inhibitory effect in fluorescent bone metastasis model of LNCaP cells compared with PC-3m cells. CONCLUSION: These results suggest that the downstream siRNA controlled by PSA promoter in retrovirus system can express selectively in androgen-responsive prostate cancer in vitro and in vivo, and CXCR4 plays an important role in prostate cancer metastasis. We believe that the pPSA-siCXCR4 retrovirus vector is a potential choice in gene therapy for androgen-responsive prostate cancer. PMID- 18431594 TI - Physiology and pathophysiology of the vasopressin-regulated renal water reabsorption. AB - To prevent dehydration, terrestrial animals and humans have developed a sensitive and versatile system to maintain their water homeostasis. In states of hypernatremia or hypovolemia, the antidiuretic hormone vasopressin (AVP) is released from the pituitary and binds its type-2 receptor in renal principal cells. This triggers an intracellular cAMP signaling cascade, which phosphorylates aquaporin-2 (AQP2) and targets the channel to the apical plasma membrane. Driven by an osmotic gradient, pro-urinary water then passes the membrane through AQP2 and leaves the cell on the basolateral side via AQP3 and AQP4 water channels. When water homeostasis is restored, AVP levels decline, and AQP2 is internalized from the plasma membrane, leaving the plasma membrane watertight again. The action of AVP is counterbalanced by several hormones like prostaglandin E2, bradykinin, dopamine, endothelin-1, acetylcholine, epidermal growth factor, and purines. Moreover, AQP2 is strongly involved in the pathophysiology of disorders characterized by renal concentrating defects, as well as conditions associated with severe water retention. This review focuses on our recent increase in understanding of the molecular mechanisms underlying AVP regulated renal water transport in both health and disease. PMID- 18431599 TI - Effect of a toxicant on phagocytosis pathways in the freshwater snail Lymnaea stagnalis. AB - The disturbance of plasma membrane carbohydrates and of lipopolysaccharide (LPS) ligands in relation to cytoskeletal transformations of haemocytes has been investigated after chronic exposure of pond snails (Lymnaea stagnalis) to the peroxidizing toxicant fomesafen. Neither of the two lectins used (concanavalin A and wheat germ agglutinin) showed any binding modification after incubation of the snails in the presence of the toxicant. However, after exposure of the snails to fomesafen, a clear and persistent reduction in LPS labelling of haemocytes occurred. The actin cytoskeleton of the same cells also appeared to be sensitive to the toxicant. The reduction in LPS-binding sites was related to actin staining, leading to the hypothesis that LPS ligands and actin could be similarly modulated by the toxicant. Damaged cells showed non-adherent membrane portions with reduced filopodial extrusions, exhibiting a smooth surface free of microvilli. These changes could lower the spreading and adhesion of the cells and could therefore account for the loss in their phagocytic capabilities. PMID- 18431598 TI - Pleiotropic effects of the yeast Sal1 and Aac2 carriers on mitochondrial function via an activity distinct from adenine nucleotide transport. AB - In Saccharomyces cerevisiae, SAL1 encodes a Ca2+ -binding mitochondrial carrier. Disruption of SAL1 is synthetically lethal with the loss of a specific function associated with the Aac2 isoform of the ATP/ADP translocase. This novel activity of Aac2 is defined as the V function (for Viability of aac2 sal1 double mutant), which is independent of the ATP/ADP exchange activity required for respiratory growth (the R function). We found that co-inactivation of SAL1 and AAC2 leads to defects in mitochondrial translation and mitochondrial DNA (mtDNA) maintenance. Additionally, sal1Delta exacerbates the respiratory deficiency and mtDNA instability of ggc1Delta, shy1Delta and mtg1Delta mutants, which are known to reduce mitochondrial protein synthesis or protein complex assembly. The V function is complemented by the human Short Ca2+ -binding Mitochondrial Carrier (SCaMC) protein, SCaMC-2, a putative ATP-Mg/Pi exchangers on the inner membrane. However, mitochondria lacking both Sal1p and Aac2p are not depleted of adenine nucleotides. The Aac2R252I and Aac2R253I variants mutated at the R252-254 triplet critical for nucleotide transport retain the V function. Likewise, Sal1p remains functionally active when the R479I and R481I mutations were introduced into the structurally equivalent R479-T480-R481 motif. Finally, we found that the naturally occurring V-R+ Aac1 isoform of adenine nucleotide translocase partially gains the V function at the expense of the R function by introducing the mutations P89L and A96 V. Thus, our data support the view that the V function is independent of adenine nucleotide transport associated with Sal1p and Aac2p and this evolutionarily conserved activity affects multiple processes in mitochondria. PMID- 18431600 TI - Remote engineering for a cheese whey biorefinery: an Internet-based application for process design, economic analysis, monitoring, and control of multiple plant sites. AB - The proteolysis of cheese whey with the aid of immobilized enzymes is an attractive alternative for this by-product of the dairy industry. Among some possible applications for whey protein hydrolysates, one may cite their use as protein source for individuals with reduced capacity of digestion, or with genetic metabolic disorders (phenylketonuria patients, for instance). The multipurpose plant that processes whey is named here as a cheese whey biorefinery. This work presents the remote control and monitoring of the whey biorefineries using the Internet. In an integrated environment, the web application also enables simulation and economic analyses of the process. This technology might allow small companies to access a remote "engineering centre", with know-how on plant design and advanced control techniques. The idea can also be extended to large dairy companies, providing the remote control of geographically spread sites of production. PMID- 18431601 TI - Hydrolysis of whey lactose using CTAB-permeabilized yeast cells. AB - Disposal of lactose in whey and whey permeates is one of the most significant problems with regard to economics and environmental impact faced by the dairy industries. The enzymatic hydrolysis of whey lactose to glucose and galactose by beta-galactosidase constitutes the basis of the most biotechnological processes currently developed to exploit the sugar content of whey. Keeping this in view, lactose hydrolysis in whey was performed using CTAB permeabilized Kluyveromyces marxianus cells. Permeabilization of K. marxianus cells in relation to beta galactosidase activity was carried out using cetyltrimethyl ammonium bromide (CTAB) to avoid the problem of enzyme extraction. Different process parameters (biomass load, pH, temperature, and incubation time) were optimized to enhance the lactose hydrolysis in whey. Maximum hydrolysis (90.5%) of whey lactose was observed with 200 mg DW yeast biomass after 90 min of incubation period at optimum pH of 6.5 and temperature of 40 degrees C. PMID- 18431603 TI - Clinical quiz: Mysterious hyperkalemia and cardiac arrest in a newborn infant undergoing continuous veno-venous hemofiltration dialysis: acute hyperkalemia. PMID- 18431604 TI - [Tuition in palliative medicine. Does it have an impact on future physicians' attitudes toward active euthanasia?]. AB - BACKGROUND: The aim of this study was to examine the attitude of medical students with or without tuition in palliative medicine towards active euthanasia and whether this changes during the course of the study period. METHODS: A questionnaire was developed with 13 items (focus groups, preliminary tests) which takes knowledge and attitude into consideration. Students in the 2nd (2CS) and 6th (6CS) clinical semesters at 2 universities with (U1) and without (U2) palliative medicine as a compulsory subject were included. RESULTS: The initial approval rate for active euthanasia was high for students in 2CS at both universities (U1 and U2), remained stable for U2 and sank clearly for U1. At U1 the number of students who would consider active euthanasia for themselves was greatly reduced but only slightly for U2. Of all the students, 40.9% of U2 and 22.5% of U1 were of the opinion that they could practice active euthanasia on patients. The majority admitted to being frightened to have responsibility for incurable patients. Of the students in 6CS, only 12.2% from U1 and 7.1% from U2 considered themselves sufficiently prepared to be responsible for terminally ill patients. CONCLUSIONS: The results of this questionnaire demonstrated a clear influence of tuition in palliative medicine on the rejection attitude of students towards active euthanasia, however, the attitude proactive euthanasia was still high. The results indicate that tuition in palliative medicine must be modified and substantially intensified. PMID- 18431605 TI - Representativity of a mesoscale network for weather-related factors governing pollen dispersal. AB - The cultivation of transgenic crops, such as maize, requires successful gene isolation in field environments. Five spatial statistical techniques are used to evaluate the use of a regional mesoscale observation network (Iowa Environmental Mesonet) as a means to drive field-scale pollen dispersion modeling. The Nearest Neighbor Index, Fractal Dimension, Morisita Index, Thiessen Polygons, and Coefficient of Representativity are computed showing the positive and negative impacts of sequential addition of observation networks into a mesonet framework (a collection of pre-existing networks). While it is shown that the arbitrary combination of disparate observing networks increases spatial resolution, this improvement is often at the expense of increased clustering due to co-location of observation sites near urban areas. Network composition in terms of density and degree of clustering was evaluated with a grid analysis using the Barnes scheme as a means to mitigate clustering and improve prediction accuracies when mesonet data are applied to modeling. This paper shows the importance of understanding and accounting for the spatial characteristics of an observational network before applying it to a modeling effort such as field scale pollen dispersion. PMID- 18431607 TI - Seventeen years of experience with reduction mammaplasty avoiding a vertical scar. AB - BACKGROUND: It is important to reshape the breast with the least scar possible. This has led us to develop a technique for large breasts where the vertical scar was eliminated. METHODS: The new nipple is positioned at 19-21 cm. The level of the new inframammary line (IML) is marked 6 cm below the new areola. All the area between the marked IML and the present sulcus is de-epithelialized. The upper flap is undermined beginning from the new IML. The tissue excision is performed medially and laterally leaving a central pedicle that also carries the nipple areola complex (NAC). The dermoglandular pedicle is shaped conically with breast shaping sutures. The NAC is passed under the upper apron flap through its new opening. RESULTS: We retrospectively reviewed 145 patients who were operated on over the past 17 years. The mean age of the patients was 41 years. The mean distance preoperatively from midclavicle to nipple was 35.3 cm. The average weight of breast tissue excised was 1073 g per side. Seventeen patients (11%) suffered from minor complications. CONCLUSION: This technique works especially well in breast reduction cases where a large mass excision is required. The horizontal scar is obvious only when the patient is lying down and cannot be seen by the patient herself. There is no risk of destroying the circular feature of the areola, which is pulled inferiorly by the vertical scar in the other techniques. PMID- 18431606 TI - The role of intraoperative cultures at the time of reimplantation in the management of infected total joint arthroplasty. AB - We reported our experience with the usefulness and interpretation of the significance of cultures at the second-stage exchange of infected arthroplasty in a prospective, 3-year follow-up study. When such intraoperative cultures were negative, patients received no therapy; when at least two cultures showed the same microorganism, results were interpreted as infection and patients were treated with antibiotics for 6-8 weeks with no more surgical procedures. Genotypic analysis (pulsed-field gel electrophoresis) was performed to analyse coagulase-negative Staphylococcus (CoNS) infections. Among 25 patients, 18 had negative cultures at the second-stage (group 1) and seven had positive cultures (group 2) receiving glycopeptides. Follow-up medians were 30 months for group 1 and 35 months for group 2; no patients in either group had persistence or recurrence of infection. All patients from group 2 had infection by CoNS at the second-stage; in six cases CoNS were also responsible for the initial infection. Genetic studies confirm that second-stage strains show different clonal identity than first-stage ones suggesting a superinfection rather than a real persistence of initial infection. Our results support the role of intraoperative cultures at the second-stage to identify patients at risk of recurrent infection who could benefit from early antibiotic therapy. The persistence of initial infections and the presence of new superinfections should be better defined according to genotypic studies. PMID- 18431608 TI - Stage-specific guided adjuvant treatment for gastric cancer. PMID- 18431609 TI - Bariatric surgery outcomes in morbidly obese with the metabolic syndrome at US academic centers. AB - BACKGROUND: The metabolic syndrome is associated with significant cardiovascular morbidity and mortality. We assessed the in-hospital outcomes of bariatric surgery in morbidly obese patients with the metabolic syndrome in comparison to a control group without the metabolic syndrome. METHODS: Using ICD-9-CM diagnosis and procedure codes, clinical data for 20,242 patients with and without the metabolic syndrome who underwent bariatric surgery over a 5-year period were obtained from the University HealthSystem Consortium database. RESULTS: The prevalence of the metabolic syndrome among bariatric surgery patients was 27.4%. Patients with the metabolic syndrome presented significantly higher overall morbidity as compared to morbidly obese patients without the metabolic syndrome (8.6% vs. 5.8%; p < 0.01), and similar mortality (0.04% vs. 0.01%; p = 0.2) after bariatric surgery. Hispanics with the metabolic syndrome had the highest morbidity rates, and men had the uppermost mortality. In-hospital bariatric surgery outcomes were significantly improved among patients who underwent laparoscopic adjustable gastric banding. CONCLUSIONS: The data suggest that the presence of the metabolic syndrome affects inter-ethnic and gender-specific short term outcomes after bariatric surgery. PMID- 18431610 TI - Regional variations in gastric bypass surgery: results from the 2005 nationwide inpatient sample. AB - BACKGROUND: The purpose of this study is to use nationally representative data to examine regional variations in the use and outcomes of gastric bypass surgery. METHODS: Using data from the Nationwide Inpatient Sample (NIS), we identified adults undergoing gastric bypass surgery (n = 92,910) in 2005. Following descriptive analyses, multiple logistic regression models were constructed to examine regional variations in the likelihood of laparoscopic vs. open approaches and in the likelihood of complications while controlling for patient and hospital characteristics. RESULTS: After indirectly adjusting for age and sex, the gastric bypass rates per 100,000 were as follows: Northeast, 70; Midwest, 39; South, 37; and West, 61. After adjusting for both patient and hospital characteristics, the odds of receiving laparoscopic surgery for patients living in the West were 1.79 times the mean [95% confidence interval (CI): 1.67-1.92], while the odds of receiving laparoscopic surgery for patients in the Midwest were 0.66 of the mean (95% CI: 0.62-0.70) and those of the Northeast were 0.88 of the mean (95% CI: 0.83-0.94). When adjusting for both patient and hospital characteristics, the odds of one or more postoperative complications among patients living in the South were greater than the mean (OR: 1.14, 95% CI: 1.02-1.26). CONCLUSIONS: Findings from this study suggest that gastric bypass surgery is more common in the Northeast and West. There is a greater likelihood of gastric bypass being performed laparoscopically in the West; it is less likely to be performed in the Northeast and Midwest. Postoperative complications are more likely to occur in the South. PMID- 18431611 TI - Serum phenylalanine concentration as a marker of liver function in obese patients before and after bariatric surgery. AB - BACKGROUND: Human obesity is associated with increased serum phenylalanine concentration, which is probably caused by liver dysfunction related to liver steatosis. This study examines whether improvements of liver function after bariatric surgery is associated with a decrease of serum phenylalanine concentration caused by an increase of phenylalanine metabolism. METHOD: Serum phenylalanine and alanine aminotransferase (an independent predictor of liver steatosis) concentrations as well as several parameters related to obesity were measured in 16 obese patients (seven men and nine women) before and 6 months after vertical banded gastroplasty. Ten (six men and four women) lean, healthy subjects served as controls. RESULTS: Obese patients before surgery had approximately twofold higher serum phenylalanine concentration than control subjects. The serum phenylalanine concentration decreased 6 months after bariatric surgery. Serum alanine aminotransferase (ALT) concentration was higher in obese patients before surgery, and decreased 6 months after bariatric surgery. Changes in serum phenylalanine concentration correlated positively with changes of ALT concentration (r = 0.75; p < 0.001). The body weight, BMI, HOMA-IR, serum triacylglycerol, LDL-cholesterol/HLD-cholesterol ratio, leptin, insulin, and glucose concentrations were higher in obese patients, and decreased 6 months after bariatric surgery. Serum HDL-cholesterol concentration was lower in obese patients before surgery than in control subjects and increased 6 months after surgery. CONCLUSION: Results obtained indicate that bariatric surgery-induced weight loss had beneficial effects on several laboratory parameters including serum phenylalanine, ALT, lipid concentrations and insulin resistance. A strong positive correlation between serum phenylalanine and serum ALT concentrations suggests that deterioration of liver function in obese patients is contributing to a decrease in phenylalanine metabolism and consequently to the increase of serum phenylalanine concentration. One can suppose that serum phenylalanine concentration could be noninvasive marker of liver dysfunction associated with liver steatosis in obese patients. PMID- 18431612 TI - Symmetrical pouch dilatation after laparoscopic adjustable gastric banding: incidence and management. AB - BACKGROUND: Laparoscopic adjustable gastric banding (LAGB) has commonly been complicated by the problem of band slippage or prolapse. Since popularization of the pars flaccida approach and improved anterior fixation, it is our impression that the problem of symmetrical dilatation of the proximal gastric pouch has become more important. METHODS: We have reviewed the results of a series of 425 LAGB all performed by the pars flaccida approach from June 2003 to October 2007 to analyze the incidence and implications of this new pattern. RESULTS: There were no posterior prolapses, 2 anterior prolapses, and 17 cases of symmetrical pouch dilatation (SPD) (revision rate 4.4%). Teenage patients had a 22% revision rate for SPD. All revisions were completed laparoscopically with no mortality, no significant complications, and a median hospital stay of 1 day. The median weight loss following revisional surgery was not significantly different from the background cohort. CONCLUSION: SPD is the most common reason for revision of LAGB in this series. We postulate that SPD is caused by excessive pressure in the proximal gastric pouch. This may be generated either by eating too quickly or too large a volume or excessive tightening of the band. The radial forces in the pouch may ultimately cause pressure on the phrenoesophageal ligament and a secondary hiatal hernia. PMID- 18431613 TI - Case reports: treatment of subtrochanteric and ipsilateral femoral neck fractures in an adult with osteopetrosis. AB - We describe a patient with autosomal-dominant osteopetrosis, a subtrochanteric fracture, and an ipsilateral femoral neck fracture treated with a hip spica cast Although the fracture united with coxa vara and external rotation deformities, the patient successfully returned to his normal activities of daily living. Operative fracture treatment in patients with osteopetrosis is difficult, and our patient provides evidence that with nonoperative treatment these patients can return to a functional level when operative treatment is not an option. PMID- 18431614 TI - Daptomycin eluted from calcium sulfate appears effective against Staphylococcus. AB - The emergence of resistant strains of Gram-positive organisms in osteomyelitis creates treatment challenges. Daptomycin is an antibiotic that shows promise for treating some resistant strains of Gram-positive infections; however, it has not been widely used clinically for the treatment of osteomyelitis. We determined whether daptomycin eluted from calcium sulfate-a local delivery vehicle used for the treatment of osteomyelitis-retained activity against Gram-positive bacteria. Daptomycin was mixed with calcium sulfate hemihydrate, with both laboratory powder and a commercial kit, to form a hardened pellet. Daptomycin was eluted from calcium sulfate and retained its ability to inhibit bacterial growth of Staphylococcus aureus and Staphylococcus epidermidis for eluates gathered up to 28 days. Our preliminary data demonstrates sterilized pellets with daptomycin retained their ability to inhibit bacterial growth of certain strains of Gram positive organisms. PMID- 18431615 TI - Papers presented at the 2007 Meeting of the Musculoskeletal Infection Society: editorial comment. PMID- 18431616 TI - Odor supported place cell model and goal navigation in rodents. AB - Experiments with rodents demonstrate that visual cues play an important role in the control of hippocampal place cells and spatial navigation. Nevertheless, rats may also rely on auditory, olfactory and somatosensory stimuli for orientation. It is also known that rats can track odors or self-generated scent marks to find a food source. Here we model odor supported place cells by using a simple feed forward network and analyze the impact of olfactory cues on place cell formation and spatial navigation. The obtained place cells are used to solve a goal navigation task by a novel mechanism based on self-marking by odor patches combined with a Q-learning algorithm. We also analyze the impact of place cell remapping on goal directed behavior when switching between two environments. We emphasize the importance of olfactory cues in place cell formation and show that the utility of environmental and self-generated olfactory cues, together with a mixed navigation strategy, improves goal directed navigation. PMID- 18431619 TI - McCloskey and me: a back-and-forth. PMID- 18431618 TI - Indirect effects of acute alcohol intoxication on sexual risk-taking: The roles of subjective and physiological sexual arousal. AB - Three experiments supported the idea that alcohol fosters sexual risk-taking in men and women, in part, through its effects on sexual arousal. In Experiment 1, increasing alcohol dosage (target blood alcohol levels of .00, .04, .08%) heightened men's and women's risk-taking intentions. Alcohol's effect was indirect via increased subjective sexual arousal; also, men exhibited greater risk-taking than women. In Experiment 2, an extended dosage range (target blood alcohol levels of .00, .06, .08, .10%) heightened men's risk-taking intentions. Alcohol's effect again was indirect via subjective arousal. Physiological sexual arousal, which was unaffected by alcohol, increased risk-taking via increased subjective arousal. In Experiment 3, alcohol increased women's risk-taking indirectly via subjective arousal, but alcohol-attenuated physiological arousal had no effect on risk-taking. Implications for alcohol myopia theory and prevention interventions are discussed. PMID- 18431620 TI - A social psychology of a history of a snippet in the psychology of transgenderism. PMID- 18431621 TI - Politics in scholarly drag: Dreger's assault on the critics of Bailey. PMID- 18431622 TI - Truth, lies, and trans science. PMID- 18431623 TI - Cowboys, sheepherders, and "The man who would be queen": "I know" vs. first-order lived experience. PMID- 18431624 TI - The Bailey affair: political correctness and attacks on sex research. PMID- 18431625 TI - Dreger's adventures. PMID- 18431626 TI - A matter of perspective: a transsexual woman-centric critique of Dreger's "scholarly history" of the Bailey controversy. PMID- 18431627 TI - A different perspective. PMID- 18431628 TI - Sex, sexuality, and gender dichotomized: transgender homosexuality in Israel. PMID- 18431629 TI - Dreger on the Bailey controversy: lost in the drama, missing the big picture. PMID- 18431630 TI - Deconstructing the feminine essence narrative. PMID- 18431631 TI - Trans victims, trans zealots: a critique of Dreger's history of the Bailey controversy. PMID- 18431632 TI - Pretenders to the throne. PMID- 18431633 TI - Shame and narcissistic rage in autogynephilic transsexualism. PMID- 18431634 TI - The drama of sex, identity, and the "queen". PMID- 18431635 TI - Lighten up, ladies. PMID- 18431636 TI - Gender identity politics, human subjects issues, and the "law of unintended consequences". PMID- 18431637 TI - Is this a work of science? PMID- 18431638 TI - Accounting for power and academic responsibility. PMID- 18431639 TI - Two modes of thought: the narrative/paradigmatic disconnect in the Bailey book controversy. PMID- 18431640 TI - Lust or identity? PMID- 18431642 TI - Introduction to Dreger (2008) and peer commentaries. PMID- 18431641 TI - The controversy surrounding "The man who would be queen": a case history of the politics of science, identity, and sex in the Internet age. AB - In 2003, psychology professor and sex researcher J. Michael Bailey published a book entitled The Man Who Would Be Queen: The Science of Gender-Bending and Transsexualism. The book's portrayal of male-to-female (MTF) transsexualism, based on a theory developed by sexologist Ray Blanchard, outraged some transgender activists. They believed the book to be typical of much of the biomedical literature on transsexuality-oppressive in both tone and claims, insulting to their senses of self, and damaging to their public identities. Some saw the book as especially dangerous because it claimed to be based on rigorous science, was published by an imprint of the National Academy of Sciences, and argued that MTF sex changes are motivated primarily by erotic interests and not by the problem of having the gender identity common to one sex in the body of the other. Dissatisfied with the option of merely criticizing the book, a small number of transwomen (particularly Lynn Conway, Andrea James, and Deirdre McCloskey) worked to try to ruin Bailey. Using published and unpublished sources as well as original interviews, this essay traces the history of the backlash against Bailey and his book. It also provides a thorough exegesis of the book's treatment of transsexuality and includes a comprehensive investigation of the merit of the charges made against Bailey that he had behaved unethically, immorally, and illegally in the production of his book. The essay closes with an epilogue that explores what has happened since 2003 to the central ideas and major players in the controversy. PMID- 18431643 TI - A TaqI PCR-RFLP detecting a novel SNP in exon 2 of the bovine POU1F1 gene. AB - PCR-SSCP and DNA sequencing methods were applied to reveal three novel single nucleotide polymorphisms (SNPs) in exon 2 of the POU1F1 gene in 963 Chinese cattle belonging to eight breeds. Among them, a silent SNP (NM_174579:c.545G > A) detected by TaqI endonuclease is described. Frequencies of the POU1F1-G allele varied from 0.685 to 1.000. The association of TaqI polymorphism with growth traits was analyzed in 251 Nanyang cattle. No significant associations of the TaqI polymorphism with body weight and average daily gain for different growth periods (6, 12, 18, and 24 months old) were observed (P > 0.05), as well as for body sizes (P > 0.05). PMID- 18431644 TI - Clinicians' self-assessment of cultural and spiritual competency: working with Asians and Asian Americans. AB - This study examines clinicians' own assessment of their cultural and spiritual competency in working with Asians and Asian Americans. Thirty clinicians, who are Asian Ethnic Minority Mental Health Specialists in the Northwest region of the United States, were surveyed to assess their perceived levels of cultural and spiritual competency. The study found that clinicians perceived themselves as being less spiritually competent than culturally competent and that most clinicians acknowledged the need for more training in spirituality. PMID- 18431645 TI - Biochemical mechanism of healing activity of the natural phenolic, allylpyrocatechol against indomethacin-induced gastric ulceration in mice. AB - Indomethacin caused maximum stomach ulceration in mice on the 3rd day, which was associated with reduction of plasma total antioxidant status (TAS), COX-1, COX-2, mucosal PGE(2), VEGF, and vWF, along with an increase in endostatin levels. Treatment with the phytochemical allylpyrocatechol (5 mg/kg, p.o. for 3 days) provided significant ulcer healing by reversing these biochemical parameters, as well as increasing the EGF expression more than that observed due to ulceration. Omeprazole (3 mg/kg, p.o. for 3 days) provided a similar healing by improving TAS and mucin levels, without significantly altering the other parameters. PMID- 18431646 TI - Effect of lyophilization and freeze-thawing on the stability of siRNA-liposome complexes. AB - The purpose of this research was to describe the application of lyophilization in the delivery of siRNA using cationic lipids by addressing the long-term formulation/stability issues associated with cationic lipids and to understand the mechanism of lyoprotection. siRNA liposomes complexes were formed in different potential cyro/lyoprotectants and subjected to either lyophilization or freeze thaw cycles. siRNA, liposomes and/or lipoplexes were tested for activity, SYBR Green I binding, cellular uptake and particle size. The lipoplexes when lyophilized in the presence of sugars as lyoprotectants could be lyophilized and reconstituted without loss of transfection efficacy but in ionic solutions they lost 65-75% of their functionality. The mechanism of this loss of activity was further investigated. The lyophilization process did not alter siRNA's intrinsic biological activity as was evident by the ability of lyophilized siRNA to retain functionality and SYBR green I binding ability. While the lipoplex size dramatically increased ( approximately 50-70 times) after lyophilization in the absence of non-ionic lyoprotectants. This increase in size correlated to the decrease in cellular accumulation of siRNA and a decrease in activity. In conclusion, siRNAs can be applied in cationic lipid lyophilized formulations and these complexes represent a potential method of increasing the stability of pre formed complex. PMID- 18431647 TI - Crystal structure determination of new antimitotic agent bis(p fluorobenzyl)trisulfide. AB - The purpose of this research was to investigate the physical characteristics and crystalline structure of bis(p-fluorobenzyl)trisulfide, a new anti-tumor agent. Methods used included X-ray single crystal diffraction, X-ray powder diffraction (XRPD), Fourier-transform infrared (FT-IR) spectroscopy, differential scanning calorimetric (DSC) and thermogravimetric (TG) analyses. The findings obtained with X-ray single crystal diffraction showed that a monoclinic unit cell was a = 12.266(1) A, b = 4.7757(4) A, c = 25.510(1) A, beta = 104.25(1) degrees ; cell volume = 1,448.4(2) A(3), Z = 4, and space group C2/c. The XRPD studies of the four crystalline samples, obtained by recrystallization from four different solvents, indicated that they had the same diffraction patterns. The diffraction pattern stimulated from the crystal structure data is in excellent agreement with the experimental results. In addition, the identical FT-IR spectra of the four crystalline samples revealed absorption bands corresponding to S-S and C-S stretching as well as the characteristic aromatic substitution. Five percent weight loss at 163.3 degrees C was observed when TG was used to study the decomposition process in the temperature range of 20-200 degrees C. DSC also allowed for the determination of onset temperatures at 60.4(1)-60.7(3) degrees C and peak temperatures at 62.1(3)-62.4(3) degrees C for the four crystalline samples studied. The results verified that the single crystal structure shared the same crystal form with the four crystalline samples investigated. PMID- 18431648 TI - Preparation and evaluation of taste masked famotidine formulation using drug/beta cyclodextrin/polymer ternary complexation approach. AB - The main aim of the present study was to evaluate potential of ternary complexation (comprising of drug, cyclodextrin and polymer) as an approach for taste masking. For this purpose famotidine with property of bitter taste was selected as a model drug. Improvement in taste masking capability of cyclodextrin towards famotidine was evaluated by formulating a ternary complex including hydrophilic polymer hydroxyl propyl methyl cellulose (HPMC 5 cps) as the third component. Phase solubility analysis at 25 degrees C was carried out for both the binary systems (viz. drug-cyclodextrin and drug-polymer) and the ternary system (drug-cyclodextrin-polymer). Ternary complex was prepared using solution method and was further characterized using XRD, DSC, FT-IR and microscopic studies. In vitro dissolution study was carried out to see the effect of ternary complexation on drug release. Taste perception study was carried out on human volunteers to evaluate the taste masking ability of ternary complexation. Results obtained from phase solubility analysis showed that the combined use of polymer and cyclodextrin effectively increased the stability constant of the complex [from 538 M(-1) for binary system to 15,096 M(-1) for ternary system]. Ternary system showed effective taste masking as compared to binary complex and at the same time showed no limiting effect on the drug release (D.E(15min) = 90%). The effective taste masking was attributed to the enhanced complexation of famotidine in ternary system compared to binary system and the same was confirmed from the characterization studies. In conclusion, the study confirmed that ternary complexation can be utilized as an alternative approach for effective taste masking. PMID- 18431649 TI - Development and characterization of zaleplon solid dispersion systems: a technical note. PMID- 18431650 TI - Pore direction in relation to anisotropy of mechanical strength in a cubic starch compact. AB - The purpose of this research was to evaluate the relation between preferential direction of pores and mechanical strength of cubic starch compacts. The preferential pore direction was quantified in SEM images of cross sections of starch compacts using a previously described algorithm for determination of the quotient of transitions (Q). This parameter and the mechanical strength were evaluated in compacts of different porosities. Starch was chosen as a model compound for materials with ductile behaviour of which tablets with low porosities can be made and which shows some elastic recovery after compaction. At medium and high porosity Q was significantly higher in the images providing a side view of the compact than in the images providing a top view (0.973 vs. 0.927 and 0.958 vs. 0.874 at 0 mm from the side of the compact and 0.956 vs. 0.854 and 0.951 vs. 0.862 at 3.5 mm), indicating that the pores were mainly oriented in the direction perpendicular to the direction of compression. This was accompanied by a lower crushing force in this direction. This could be explained by considering the pores as cracks which propagate through the sample during crushing. For both directions the crushing force decreased with increasing porosity. The yield strength of the compacts also decreased with increasing porosity, but this parameter was not dependent on the direction of crushing when the porosity was below 10%. The results show that pore direction significantly influences the crushing force but does not influence the yield strength, at porosities below 10%. PMID- 18431651 TI - A review of current intravaginal drug delivery approaches employed for the prophylaxis of HIV/AIDS and prevention of sexually transmitted infections. AB - The objective of this review is to describe the current status of several intravaginal anti-HIV microbicidal delivery systems these delivery systems and microbicidal compounds in the context of their stage within clinical trials and their potential cervicovaginal defence successes. The global Human Immuno Deficiency Virus (HIV) pandemic continues to spread at a rate of more than 15,000 new infections daily and sexually transmitted infections (STIs) can predispose people to acquiring HIV infection. Male-to-female transmission is eight times more likely to occur than female-to-male transmission due to the anatomical structure of the vagina as well as socio-economic factors and the disempowerment of women that renders them unable to refuse unsafe sexual practices in some communities. The increased incidence of HIV in women has identified the urgent need for efficacious and safe intravaginal delivery of anti-HIV agents that can be used and controlled by women. To meet this challenge, several intravaginal anti-HIV microbicidal delivery systems are in the process of been developed. The outcomes of three main categories are discussed in this review: namely, dual function polymeric systems, non-polymeric systems and nanotechnology-based systems. These delivery systems include formulations that modify the genital environment (e.g. polyacrylic acid gels and lactobacillus gels), surfactants (e.g. sodium lauryl sulfate), polyanionic therapeutic polymers (e.g. carageenan and carbomer/lactic acid gels), proteins (e.g. cyanovirin-N, monoclonal antibodies and thromspondin-1 peptides), protease inhibitors and other molecules (e.g. dendrimer based-gels and the molecular condom). Intravaginal microbicide delivery systems are providing a new option for preventing the transmission of STIs and HIV. PMID- 18431653 TI - Effect of drug lipophilicity and ionization on permeability across the buccal mucosa: a technical note. PMID- 18431652 TI - Embryonic chicken trachea as a new in vitro model for the investigation of mucociliary particle clearance in the airways. AB - Mucociliary clearance (MC) is an important defense mechanism of the respiratory system to eliminate inhaled and possibly noxious particles from the lung. Although the principal mechanics of MC seem to be relatively clear there are still open questions regarding the long-term clearance of particles. Therefore, we have developed a new set-up based on embryonic chicken trachea (ECT) to investigate mucociliary particle clearance in more detail. ECT was placed in an incubation chamber after carbon particles were applied and tracked using optical microscopy. The aim of the study was to validate this model by investigating the impact of temperature, humidity and drugs on particle transport rates. Particles were transported reproducibly along the trachea and clearance velocity (2.39 +/- 0.25) mm/min was found to be in accordance to data reported in literature. Variation in temperature resulted in significantly reduced MC: (0.40 +/- 0.12) mm/min (20 degrees C); (0.42 +/- 0.10) mm/min (45 degrees C). Decreasing humidity (99-60%) had no significant effect on MC, whereas reduction to 20% humidity showed a significant influence on particle clearance. The use of different cilio- and muco-active drugs (propranolol, terbutalin, N-acetylcysteine) resulted in altered MC according to the pharmacological effect of the substances: a concentration dependent decrease of MC was found for Propranolol. From our results we conclude that this model can be employed to investigate MC of particles in more detail. Hence, the model may help to understand and identify decisive physico-chemical parameters for MC and to answer open questions regarding the long-term clearance phenomenon. PMID- 18431654 TI - Pluronic and tetronic copolymers with polyglycolyzed oils as self-emulsifying drug delivery systems. AB - The potential of poly(ethylene oxide)-poly(propylene oxide) block copolymers Pluronic F127 (PF127) and Tetronic 304 (T304), 904 (T904) and 1307 (T1307) as components of solid self-(micro)emulsifying dosage forms, S(M)EDDS, was evaluated. The dependence of the self-associative properties of Tetronics on pH explained the low ability of the micelles to solubilize griseofulvin at acid pH (sevenfold increase) compared to at alkaline pH (12-fold). Blends of polyglycolyzed glycerides (Labrasol, Labrafac CC, and Labrafil M 1944CS) with each copolymer at two different weight ratios (80:20 and 60:40) were prepared, diluted in water, and characterized in terms of globule size, appearance and griseofulvin solubility. The blends with Labrasol led to microemulsions that are able to increase drug solubility up to 30-fold. SMEDD hard gelatine capsules filled with griseofulvin and Labrasol or Labrasol/copolymer 80:20 showed a remarkable increase in drug solubility and dissolution rate, particularly when T904, T1307 or PF127 was present in the blend. This effect was more remarkable when the volume of the dissolution medium was 200 ml (compared to 900 ml), which can be related to a higher stability of the microemulsion when there is a greater concentration of the copolymer and glyceride in the medium. PMID- 18431655 TI - Preformulation considerations for controlled release dosage forms. Part I. Selecting candidates. AB - The physical-chemical properties of interest for controlled release (CR) dosage form development presented are based on the author's experience. Part I addresses selection of the final form based on a logical progression of physical-chemical properties evaluation of candidate forms and elimination of forms with undesirable properties from further evaluation in order to simplify final form selection. Several candidate forms which could include salt, free base or acid, polymorphic and amorphic forms of a new chemical entity (NCE) or existing drug substance (DS) are prepared and evaluated for critical properties in a scheme relevant to manufacturing processes, predictive of problems, requiring small amounts of test materials and simple analytical tools. A stability indicating assay is not needed to initiate the evaluation. This process is applicable to CR and immediate release (IR) dosage form development. The critical properties evaluated are melting, crystallinity, solubilities in water, 0.1 N HCl, and SIF, hygrodymamics, i.e., moisture sorption and loss at extremes of RH, and LOD at typical wet granulation drying conditions, and processability, i.e., corrosivity, and filming and/or sticking upon compression. PMID- 18431656 TI - Preformulation considerations for controlled release dosage forms. Part II. Selected candidate support. AB - Practical examples of preformulation support of the form selected for formulation development are provided using several drug substances (DSs). The examples include determination of the solubilities vs. pH particularly for the range pH 1 to 8 because of its relationship to gastrointestinal (GI) conditions and dissolution method development. The advantages of equilibrium solubility and trial solubility methods are described. The equilibrium method is related to detecting polymorphism and the trial solubility method, to simplifying difficult solubility problems. An example of two polymorphs existing in mixtures of DS is presented in which one of the forms is very unstable. Accelerating stability studies are used in conjunction with HPLC and quantitative X-ray powder diffraction (QXRD) to demonstrate the differences in chemical and polymorphic stabilities. The results from two model excipient compatibility methods are compared to determine which has better predictive accuracy for room temperature stability. A DSC (calorimetric) method and an isothermal stress with quantitative analysis (ISQA) method that simulates wet granulation conditions were compared using a 2 year room temperature sample set as reference. An example of a pH stability profile for understanding stability and extrapolating stability to other environments is provided. The pH-stability of omeprazole and lansoprazole, which are extremely unstable in acidic and even mildly acidic conditions, are related to the formulation of delayed release dosage forms and the resolution of the problem associated with free carboxyl groups from the enteric coating polymers reacting with the DSs. Dissolution method requirements for CR dosage forms are discussed. The applicability of a modified disintegration time (DT) apparatus for supporting CR dosage form development of a pH sensitive DS at a specific pH such as duodenal pH 5.6 is related. This method is applicable for DSs such as peptides, proteins, enzymes and natural products where physical observation can be used in place of a difficult to perform analytical method, saving resources and providing rapid preformulation support. PMID- 18431657 TI - Development and evaluation of artemether taste masked rapid disintegrating tablets with improved dissolution using solid dispersion technique. AB - The purpose of this research was to mask the intensely bitter taste of artemether (ARM) and to formulate a rapid-disintegrating tablet (RDT) of the taste-masked drug. Taste masking was done by solid dispersion with mono amino glycyrrhyzinate pentahydrate (GLY) by solvent evaporation method. To characterize and formulate taste masked rapid disintegrating tablets (RDTs) of ARM, the 1:1M solid dispersion was selected based on bitterness score. Fourier transform infrared spectroscopy (FTIR), differential scanning calorimetry (DSC), and X-ray powder diffraction (XRPD) were performed to identify the physicochemical interaction between drug and carrier, hence its effect on dissolution. RDTs were evaluated for weight variation, disintegration time, hardness and friability. In vitro drug release studies were performed for RDTs at pH 1.2 and 6.8. Bitterness score was evaluated using mini-column method and compared with gustatory sensation test. FTIR spectroscopy and DSC showed no interaction while XRPD showed amorphization of ARM in GLY solid dispersion. RDTs prepared using solid dispersion, (RDT3), showed faster disintegration (within 28 s) and complete bitter taste masking of ARM. In addition, RDT3 exhibited better dissolution profile at both pH 1.2 and 6.8, than RDTs prepared from pure ARM (RDT5). Taste evaluation of RDTs in human volunteers rated tasteless with a score of 0 to RDT3 and 3 to RDT5. Mini-column revealed that RDT5 showed increase in number of persons who sensed bitterness with increased amount of ARM release while RDT3 sensed no bitterness. Thus, results conclusively demonstrated successful masking of taste and rapid disintegration of the formulated tablets in the oral cavity with improved dissolution. PMID- 18431658 TI - Floating osmotic drug delivery system of ranitidine hydrochloride: development and evaluation--a technical note. PMID- 18431659 TI - Effect of surfactants and solutes (glucose and NaCl) on solubility of Kavain--a technical note. PMID- 18431660 TI - A novel nanoparticle formulation for sustained paclitaxel delivery. AB - PURPOSE: To develop a novel nanoparticle drug delivery system consisting of chitosan and glyceryl monooleate (GMO) for the delivery of a wide variety of therapeutics including paclitaxel. METHODS: Chitosan/GMO nanoparticles were prepared by multiple emulsion (o/w/o) solvent evaporation methods. Particle size and surface charge were determined. The morphological characteristics and cellular adhesion were evaluated with surface or transmission electron microscopy methods. The drug loading, encapsulation efficiency, in vitro release and cellular uptake were determined using HPLC methods. The safety and efficacy were evaluated by MTT cytotoxicity assay in human breast cancer cells (MDA-MB-231). RESULTS: These studies provide conceptual proof that chitosan/GMO can form polycationic nano-sized particles (400 to 700 nm). The formulation demonstrates high yields (98 to 100%) and similar entrapment efficiencies. The lyophilized powder can be stored and easily be resuspended in an aqueous matrix. The nanoparticles have a hydrophobic inner-core with a hydrophilic coating that exhibits a significant positive charge and sustained release characteristics. This novel nanoparticle formulation shows evidence of mucoadhesive properties; a fourfold increased cellular uptake and a 1000-fold reduction in the IC(50) of PTX. CONCLUSION: These advantages allow lower doses of PTX to achieve a therapeutic effect, thus presumably minimizing the adverse side effects. PMID- 18431661 TI - Preparation and evaluation of diltiazem hydrochloride diffusion-controlled transdermal delivery system. AB - The objective was to investigate the suitable polymeric films for the development of diltiazem hydrochloride (diltiazem HCl) transdermal drug delivery systems. Hydroxypropyl methylcellulose (HPMC) and ethylcellulose (EC) were used as hydrophilic and hydrophobic film formers, respectively. Effects of HPMC/EC ratios and plasticizers on mechanical properties of free films were studied. Effects of HPMC/EC ratios on moisture uptake, in vitro release and permeation through pig ear skin of diltiazem HCl films were evaluated. Influence of enhancers including isopropyl myristate (IPM), isopropyl palmitate (IPP), N-methyl-2-pyrrolidone, oleic acid, polyethylene glycol 400, propylene glycol, and Tween80 on permeation was evaluated. It was found that addition of EC into HPMC film produced lower ultimate tensile strength, percent elongation at break and Young's modulus, however, addition of EC up to 60% resulted in too hard film. Plasticization with dibutyl phthalate (DBP) produced higher strength but lower elongation as compared to triethyl citrate. The moisture uptake and initial release rates (0-1 h) of diltiazem HCl films decreased with increasing the EC ratio. Diltiazem HCl films (10:0, 8:2 and 6:4 HPMC/EC) were studied for permeation because of the higher release rate. The 10:0 and 8:2 HPMC/EC films showed the comparable permeation time profiles, and had higher flux values and shorter lag time as compared to 6:4 HPMC/EC film. Addition of IPM, IPP or Tween80 could enhance the fluxes for approx. three times while Tween80 also shorten the lag time. In conclusion, the film composed of 8:2 HPMC/EC, 30% DBP and 10% IPM, IPP or Tween80 loaded with 25% diltiazem HCl should be selected for manufacturing transdermal patch by using a suitable adhesive layer and backing membrane. Further in vitro permeation and in vivo performance studies are required. PMID- 18431662 TI - Physical characterization of a new skin bioadhesive film. AB - Physical properties (roughness, gloss, mechanical, surface topography and adhesive) of a bioadhesive film for the transdermal delivery of drugs and its interactions with a skin model surface were studied. Roughness is a measurement of the small-scale variations in the height of a physical surface. No significant differences in Ra between the "x" and "y" dimensions for both the skin model and patch were detected, due to uniformity in their production. Scanning electron microscope pictures showed small particles projected from the film. Those particles resulted in increasing roughness and surface area. For the patch, gloss values measured at 20 degrees were 6.0 +/- 0.9 and at 60 degrees , 32.2 +/- 2.2 gloss units, respectively, indicating a semi-gloss material. Concerning the mechanical properties, the tensile strength of the film resulted four- to sevenfold greater than the peel force from the model skin used, indicating the suitability of the film for skin application. The adhesion to skin model depended on the amount of water used for film application and on the elapsed time between film application and removal. Finally, the model skin that was invented by Charkoudian can be used as an alternative to costly and highly variable human skin substrates since it possesses human topography. PMID- 18431663 TI - Effect of drug solubility on polymer hydration and drug dissolution from polyethylene oxide (PEO) matrix tablets. AB - The purpose of the study was to investigate the effect of drug solubility on polymer hydration and drug dissolution from modified release matrix tablets of polyethylene oxide (PEO). Different PEO matrix tablets were prepared using acetaminophen (ACE) and ibuprofen (IBU) as study compounds and Polyox WSR301 (PEO) as primary hydrophilic matrix polymer. Tablet dissolution was tested using the USP Apparatus II, and the hydration of PEO polymer during dissolution was recorded using a texture analyzer. Drug dissolution from the preparations was dependent upon drug solubility, hydrogel formation and polymer proportion in the preparation. Delayed drug release was attributed to the formation of hydrogel layer on the surface of the tablet and the penetration of water into matrix core through drug dissolution and diffusion. A multiple linear regression model could be used to describe the relationship among drug dissolution, polymer ratio, hydrogel formation and drug solubility; the mathematical correlation was also proven to be valid and adaptable to a series of study compounds. The developed methodology would be beneficial to formulation scientists in dosage form design and optimization. PMID- 18431664 TI - Effect of particle size on compaction of materials with different deformation mechanisms with and without lubricants. AB - This work investigates the effect of excipient particle size on compaction properties of brittle, plastic and viscoelastic materials with and without added lubricants. Sieve cuts of microcrystalline cellulose (MCC), starch and dibasic calcium phosphate dihydrate were obtained by sieving, then samples were tested without lubrication or with added lubricant (0.5% Mg stearate mixed for either 5 or 30-min). Compacts were left overnight before testing. It was found that in the absence of lubricant, compact tensile strength (TS) was dependent on particle size only for starch. With Mg stearate, lubricant sensitivity shows a strong dependence on excipient particle size for both starch and MCC, where smaller particles are less affected by lubricant. Dibasic calcium phosphate dihydrate was not sensitive to lubricant even after 30 min mixing. This study highlights that in the absence of lubricant, initial particle size of excipients has no impact on compact strength not only for dibasic calcium phosphate dihydrate (brittle), but also for MCC (plastic). On the other hand, TS is dependent on particle size both with or without added lubricant for starch (viscoelastic). PMID- 18431665 TI - Investigation of the drug release and surface morphological properties of film coated pellets, and physical, thermal and mechanical properties of free films as a function of various curing conditions. AB - The purpose of the present investigation was to elucidate the influence of curing on different physical properties of Eudragit NE and RS coating systems. Increased curing times resulted in decreased drug release rates from Eudragit NE-coated beads. However, an increase in drug release rates was noticed at longer curing times and higher temperatures for the Eudragit RS coating system. The surface morphological changes of the film-coated beads revealed that there were no visible macroscopic changes as a result of curing. The absence of any ibuprofen melting peak in the DSC thermograms of cured NE and RS coated beads confirmed that there was no surface crystallization of ibuprofen. These results indicated that the increase in drug release rates from RS coated pellets, when subjected to long curing times, resulted from loss of plasticizer. Free films of Eudragit NE exhibited an increase in tensile strength with increased curing times, whereas Eudragit RS free films showed a decrease in tensile strength beyond 4 h of curing at 70 and 90 degrees C. The film thicknesses and weights of free films of Eudragit RS prepared with triethyl citrate plasticizer were found to change more dramatically with curing than did free films of Eudragit RS prepared with ibuprofen as the plasticizer. An increase in pore volume was also observed with increased curing times for Eudragit RS free films. Such changes with curing were shown to be due to the loss of plasticizer molecules, leading to the formation of molecular-scale voids and channels. PMID- 18431666 TI - Micellar solubilization of some poorly soluble antidiabetic drugs: a technical note. PMID- 18431667 TI - Carvedilol: solubilization and cyclodextrin complexation: a technical note. PMID- 18431668 TI - Investigation of the variability of NIR in-line monitoring of roller compaction process by using Fast Fourier Transform (FFT) analysis. AB - The purpose of this research was to investigate the variability of the roller compaction process while monitoring in-line with near-infrared (NIR) spectroscopy. In this paper, a pragmatic method in determining this variability of in-line NIR monitoring roller compaction process was developed and the variability limits were established. Fast Fourier Transform (FFT) analysis was used to study the source of the systematic fluctuations of the NIR spectra. An off-line variability analysis method was developed as well to simulate the in line monitoring process in order to determine the variability limits of the roller compaction process. For this study, a binary formulation was prepared composed of acetaminophen and microcrystalline cellulose. Different roller compaction parameters such as roll speed and feeding rates were investigated to understand the variability of the process. The best-fit line slope of NIR spectra exhibited frequency dependence only on the roll speed regardless of the feeding rates. The eccentricity of the rolling motion of rollers was identified as the major source of variability and correlated with the fluctuations of the slopes of NIR spectra. The off-line static and dynamic analyses of the compacts defined two different variability of the roller compaction; the variability limits were established. These findings were proved critical in the optimization of the experimental setup of the roller compaction process by minimizing the variability of NIR in-line monitoring. PMID- 18431669 TI - Permeation studies of indomethacin from different emulsions for nasal delivery and their possible anti-inflammatory effects. AB - The purpose of this research was to develop an emulsion formulation of indomethacin (IND) suitable for nasal delivery. IND was incorporated into the oil phases of oil in water (O/W) and water in oil (W/O) emulsions. For this purpose, different emulsifying agents (Tween 80, Span 80 and Brij 58) were used in two emulsion formulations. When the effects of several synthetic membranes (nylon, cellulose, cellulose nitrate) were compared with the sheep nasal mucosa, the cellulose membrane and sheep nasal mucosa showed similar permeation properties for O/W emulsion (P > 0.05). To examine the absorption characteristics of IND, the anti-inflammatory properties of intravenous solution of IND, intranasal O/W emulsions of IND (with or without enhancers) and intranasal solution of IND (IND Sol) were investigated in rats with carrageenan-induced paw edema. When citric acid was added to the nasal emulsion, the anti-inflammatory activity was similar to that of intravenous solution (P > 0.05). Finally, it was concluded that, intranasal administration of IND emulsion with citric acid may be considered as an alternative to intravenous and per oral administrations of IND to overcome their adverse effects. PMID- 18431670 TI - Design and optimization of mefloquine hydrochloride microparticles for bitter taste masking. AB - The objective of the present investigation was to reduce the bitterness with improved dissolution, in acidic medium (pH 1.2), of mefloquine hydrochloride (MFL). Microparticles were prepared by coacervation method using Eudragit E (EE) as polymer and sodium hydroxide as precipitant. A 3(2) full factorial design was used for optimization wherein the drug concentration (A) and polymer concentration (B) were selected as independent variables and the bitterness score, particle size and dissolution at various pH were selected as the dependent variables. The desirability function approach has been employed in order to find the best compromise between the different experimental responses. The model is further cross validated for bias. The optimized microparticles were characterized by FT-IR, DSC, XRPD and SEM. Bitterness score was evaluated by human gustatory sensation test. Multiple linear regression analysis revealed that the reduced bitterness of MFL can be obtained by controlling the dissolution of microparticles at pH 6.8 and increasing the EE concentration. The increase in polymer concentration leads to reduction in dissolution of microparticles at pH > 5 due to its insolubility. However the dissolution studies at pH 1.2 demonstrated enhanced dissolution of MFL from microparticles might be due to the high porosity of the microparticles, hydrophilic nature of the EE, and improved wettability, provided by the dissolved EE. The bitterness score of microparticles was decreased to zero compared to 3+ of pure ARM. In conclusion the bitterness of MFL was reduced with improved dissolution at acidic pH. PMID- 18431671 TI - Solubility improvement of drugs using N-methyl pyrrolidone. AB - The solubilization efficiency of N-methyl pyrrolidone (NMP) has been determined and compared to that of ethanol and propylene glycol for 13 poorly soluble drugs. NMP is found to be a more efficient solubilizer for all the drugs studied. The solubility enhancement as high as about 800-fold is obtained in 20% v/v NMP solution as compared to water. The mechanism of drug solubilization by NMP has also been investigated. It is proposed that NMP enhances drug solubility by simultaneously acting as a cosolvent and a complexing agent. A mathematical model is used to estimate the drug solubility in NMP-water mixture, according to which the total solubility enhancement is a sum of the two effects. This model describes the experimental data well and is more accurate than other models. A large and uniform reduction in the surface tension of water as a function of NMP concentration demonstrates its cosolvent effect. The complexation is supported by the fact that it's strength is affected by the temperature and the polarity of the medium. A strong correlation exists between log K (ow) of the drugs and the cosolvency coefficients. The correlation between log K (ow) and the complexation coefficients is weak suggesting that factors such as molecular shape and aromaticity of the drug molecule are significant in determining the complexation strength. This has been confirmed by the absence of a significant complexation between NMP and linear drug-like solutes. PMID- 18431672 TI - Visualizing solvent mediated phase transformation behavior of carbamazepine polymorphs by principal component analysis. AB - The purpose of this research is to gain a greater insight into the hydrate formation processes of different carbamazepine (CBZ) anhydrate forms in aqueous suspension, where principal component analysis (PCA) was applied for data analysis. The capability of PCA to visualize and to reveal simplified structures that often underlie large data sets are explored. Different CBZ polymorphs were dispersed separately in aqueous solution, and then recovered and measured by FT Raman spectroscopy. PCA was employed for visualizing the dynamics of the phase transformation from each CBZ polymorph to the dihydrate (DH). As a comparison to PCA visualization, the transformation process of each CBZ polymorph was quantified using PLS modeling. The results demonstrated that PCA has advantages in presenting the original data in terms of the differences and similarities, and also directly identify the statistical patterns in the data even when the data set is large. These advantages provided greater insight into the measured Raman spectra as well as the phase transformation process of CBZ polymorphs to the DH in aqueous environment. PMID- 18431673 TI - Synthesis and characterization of pectin/PVP hydrogel membranes for drug delivery system. AB - The purpose of the present study was to develop and design pectin and polyvinyl pyrrolidone (PVP) blended hydrogel membranes (PEVP), with different pectin: PVP ratios (1:0.2, 1:0.4, 1:0.6, 1:0.8 and 1:1 w/w), which were prepared by using a conventional solution casting technique. An attempt has been made to characterize the hydrogel membranes by various instrumental techniques like, FTIR (Fourier transform infrared) spectroscopy, X-ray diffraction (XRD), Differential scanning calorimetry (DSC), tensile strength test and scanning electron microscopy (SEM). The release patterns of the drug (salicylic acid) from the hydrogel membrane were done in three different release mediums (pH 1.4, pH 7.4 and distilled water) and samples were analyzed spectrophotometrically at 294 nm wavelength on a UV Vis spectrophotometer. MTT assay was done to ensure cytocompatibility of the pectin/PVP hydrogel membranes using B16 melanoma cells. FTIR spectroscopy indicated the presence of secondary amide (I) absorption bands. The XRD study shows decrease in crystallinity of the hydrogel membranes with increase in PVP ratio. DSC study shows an increase in T(g) of pectin after blending with PVP. It was found that tensile strength increases with increasing PVP ratios in the hydrogel membranes. The prepared hydrogel membranes were found to be biocompatible with B16 melanoma cells. PMID- 18431674 TI - Formulation and evaluation of fast dissolving films for delivery of triclosan to the oral cavity. AB - The present investigation was undertaken with the objective of formulating TC containing fast dissolving films for local delivery to oral cavity. Various film forming agents, film modifiers and polyhydric alcohols were evaluated for optimizing the composition of fast dissolving films. The potential of poloxamer 407 and hydroxypropyl-beta- cyclodextrin (HPBCD) to improve solubility of TC was investigated. Fast dissolving films containing hydroxypropyl methylcellulose (HPMC), xanthan gum, and xylitol were formulated. Use of poloxamer 407 and HPBCD resulted in significant improvement in the solubility of TC. Fast dissolving films containing TC-HPBCD complex and TC-Poloxamer 407 were formulated and were evaluated for the in vitro dissolution profile and in vitro microbiological assay. Films containing TC-Poloxamer 407 exhibited better in vitro dissolution profile and in vitro antimicrobial activity as compared to the films containing TC-HPBCD complex. Effect of incorporation of eugenol on the in vivo performance of TC-Poloxamer 407 containing films was evaluated in human volunteers. Eugenol containing films improved the acceptability of TC-Poloxamer 407 films with respect to taste masking and mouth freshening without compromising the in vivo dissolution time. PMID- 18431675 TI - Minimizing variability of cascade impaction measurements in inhalers and nebulizers. AB - The purpose of this article is to catalogue in a systematic way the available information about factors that may influence the outcome and variability of cascade impactor (CI) measurements of pharmaceutical aerosols for inhalation, such as those obtained from metered dose inhalers (MDIs), dry powder inhalers (DPIs) or products for nebulization; and to suggest ways to minimize the influence of such factors. To accomplish this task, the authors constructed a cause-and-effect Ishikawa diagram for a CI measurement and considered the influence of each root cause based on industry experience and thorough literature review. The results illustrate the intricate network of underlying causes of CI variability, with the potential for several multi-way statistical interactions. It was also found that significantly more quantitative information exists about impactor-related causes than about operator-derived influences, the contribution of drug assay methodology and product-related causes, suggesting a need for further research in those areas. The understanding and awareness of all these factors should aid in the development of optimized CI methods and appropriate quality control measures for aerodynamic particle size distribution (APSD) of pharmaceutical aerosols, in line with the current regulatory initiatives involving quality-by-design (QbD). PMID- 18431676 TI - Comparative in vitro study of six carbamazepine products. AB - The purpose of present study was to evaluate commercial preparations of carbamazepine tablets with respect to drug release through a defined sequence of experiments using Minitab software. The compliance of products with respect to United States Pharmacopeia (USP) dissolution test and comparison of the products with respect to drug release in different dissolution conditions is reported in the present paper. The different dissolution conditions studied include dissolution medium (1% SLS in purified water, 0.1 N HCl), volume (900 and 1,000 ml), rpm (50 rpm, 75 rpm). Studies indicated that all six products complied with USP dissolution criteria. However, the extent of influence of dissolution conditions on drug release was varied among the products. Distinct dissolution profiles were observed and there was no correlation with disintegration time in certain products. The in vitro dissolution experimentation helped in identifying the discriminatory dissolution conditions and also the formulations that were unaffected with change of dissolution variables. In summary, commercial preparations of carbamazepine vary widely in their dissolution behavior in multi dissolution run experimentation. Identifying this behavior of the products was essential as an in vitro tool for screening a good and a bad formulation. PMID- 18431677 TI - Cellulosic ethanol production using the naturally occurring xylose-fermenting yeast, Pichia stipitis. AB - Rising crude oil prices and environmental concerns have renewed interest in renewable energy. Cellulosic ethanol promises to deliver a renewable fuel from non-food feedstocks. One technical challenge producing cellulosic ethanol economically is a robust organism to utilize the different sugars present in cellulosic biomass. Unlike starch where glucose is the only sugar present, cellulosic biomass has other sugars such as xylose and arabinose, usually called C5 sugars. This review examines the most promising naturally occurring C5 fermenting organism, Pichia stipitis. In this work, the properties that make P. stipitis unique from other organisms, its physiology and fermentation results on lignocellulosic substrates have been reviewed. P. stipitis can produce 41 g ethanol/l with a potential to cleanup some of the most concentrated toxins. These results coupled with the less stringent nutritional requirements, great resistance to contamination and its thick cell walls makes P. stipitis a viable organism for scale-up. However, P. stipitis has a slower sugar consumption rate compared to Saccharomyces cerevisiae and requires microaerophilic condition for ethanol production. Finally, future studies to enhance fermentation capabilities of this yeast have been discussed. PMID- 18431678 TI - Vanadium-enriched chickpea sprout ameliorated hyperglycemia and impaired memory in streptozotocin-induced diabetes rats. AB - Vanadium compounds have been recognized for their hypoglycemic effects; however, potential short and long-term vanadium toxicity has slowed the acceptance for therapeutic use. In the present work, three batches of vanadium-enriched chickpea sprout (VCS) were prepared by incubating chickpea seeds in presence of 200, 100, and 50 microg/ml of sodium orthovanadate (SOV). The effects of oral administration of chickpea sprout (CS) and VCS food for 8 weeks on streptozotocin induced (STZ) diabetic rats were investigated. Both CS and VCS food was found to ameliorate some hyperglycemic symptoms of the diabetic rats, i.e. improve lipid metabolism, decrease blood glucose level, prevent body weight loss, and reduce impairment of diabetic related spatial learning and memory. Serum insulin was substantially elevated in treated diabetic rats, which is probably one important reason for the hypoglycemic effect. Compared with CS alone, VCS100 food exhibited remarkably enhanced effectiveness in alleviating diabetes induced hyperglycemia and memory loss. Moreover, vanadium-enriched chickpeas appeared to abolish the vanadium induced toxicity associated with administration of this metal for diabetes during the 8-week study period. This study suggested further work of the vanadium speciation in CS and novel hypoglycemic mechanism for the antidiabetic activity of vanadium agents. Vanadium containing (VCS) food could be a dietary supplement for the diabetic status. PMID- 18431679 TI - Genetic diversity and gene flow in a Caribbean tree Pterocarpus officinalis Jacq.: a study based on chloroplast and nuclear microsatellites. AB - We analysed the molecular diversity of Pterocarpus officinalis, a tree species distributed in Caribbean islands, South and Central America to quantify the genetic variation within island, to assess the pattern of differentiation and infer levels of gene flow; with the overall goal of defining a strategy of conservation. Two hundred two individuals of 9 populations were analysed using three chloroplast and six nuclear microsatellite markers. The observed heterozygosity varied markedly among the populations for nuclear (H(Onuc )= 0.20 0.50) and chloroplast microsatellites (H (cp )= 0.22-0.68). The continental population from French Guyana showed a higher value of H(Onuc) than island populations, and the differences were significant in some cases. The fixation index F (IS) ranged from -0.043 to 0.368; a significant heterozygote deficit was detected in 7 populations. The heterozygosity excess method suggested that two populations in Guadeloupe have undergone a recent bottleneck. Global and pairwise F (ST) were high for both nuclear (F(STnuc )= 0.29) and chloroplast microsatellites (F(STcp )= 0.58). The neighbour-joining tree based on both markers, presented a differentiation pattern that can be explained by the seed dispersal by flotation and marine stream. The comparison of Bayesian approach and the method based on allelic frequency demonstrate a very limited number of migrants between populations. PMID- 18431680 TI - Social inequalities in the incidence and case fatality of cancers of the lung, the stomach, the bowels, and the breast. AB - OBJECTIVE: In order to examine health inequalities in terms of incidences and case fatalities in a German health insurance population. Lung cancer, stomach cancer, intestinal carcinoma, and breast cancer were considered. Social differentiation was depicted by income and occupational position in order to examine which one is more strongly associated with incidence and case fatality. METHODS: Analyses were performed using data from a statutory health insurance (n = 170,848). Incomes were divided into quintiles, and subjects were grouped according to occupational status. RESULTS: For lung cancer incidence a gradient between the highest and the lowest 20% of the income distribution emerged. The relative risk of the lowest category was RR = 7.03, for occupational position the figure was RR = 6.98. For stomach cancer the relative risks were RR = 5.33 for income and RR = 7.11 for occupational position. For intestinal carcinoma only income was significantly related with incidence (RR = 4.37 for the lowest 20% of the income distribution), and for breast cancer incidence no social inequalities were found. For case fatality increased relative risks emerged for lung cancer, but only for income. CONCLUSIONS: Income and occupational position were associated with cancer incidence with the exception of breast cancer. Apart from lung cancer, case fatalities were unrelated to measures of social differentiation. PMID- 18431681 TI - Location and expression of ribosomal RNA genes in grasshoppers: abundance of silent and cryptic loci. AB - We investigate regularities and restrictions in chromosome location of ribosomal RNA genes, analysed by fluorescent in situ hybridization (FISH), and their phenotypic expression assessed by nucleolus formation at first meiotic prophase cells, analysed by silver impregnation, in 49 grasshopper species. High variation was found for rDNA location between species within most genera analysed. The mean haploid number of rDNA loci detected by FISH was 2.47, but some species had up to 10 loci. Chromosome distribution of rDNA loci differed between the Gomphocerinae and Oedipodinae subfamilies, most loci being proximal to the centromere in the former and distal to it in the latter. Chromosomes 2, 3 and X frequently carried rDNA in Gomphocerinae species with 2n male symbol=17 chromosomes, whereas chromosomes 6 and 9 were the most frequent rDNA locations in the Oedipodinae. About 13% of the 126 rDNA loci detected by FISH were silent, although this figure might be even higher. The comparison of FISH and silver-impregnation results also suggested the existence of cryptic NORs, i.e. those forming small nucleoli with no apparent presence of rDNA revealed by FISH. This was especially clear after the same cells in two species were sequentially treated with both silver impregnation and FISH. The abundance of silent and cryptic loci might thus suggest that rDNA spreads through grasshopper genomes by the Dubcovsky and Dvorak mechanism-that is, the transposition of a few rRNA genes to new chromosome locations, their amplification giving rise to new NORs, and the elimination of the old NORs. The cryptic NORs might correspond to nascent NORs, i.e. a few rRNA gene copies moved to new locations, whereas the inactive rDNA loci might correspond to those being in the process of elimination. PMID- 18431682 TI - Cytogenetic analysis of the Asian plethodontid salamander, Karsenia koreana: evidence for karyotypic conservation, chromosome repatterning, and genome size evolution. AB - A cytogenetic analysis, including the karyotype, C-bands, silver-stained nucleolus organizer regions and genome size, was performed on the recently discovered species, Karsenia koreana, the first plethodontid salamander from Asia. The karyotype consists of 14 pairs of bi-armed chromosomes, with no evidence of heteromorphic sex chromosomes. C-banding reveals a concentration of heterochromatin at the centromeres as well as at interstitial locations. The smallest chromosome (pair number 14) has symmetrical interstitial C-bands in each arm, resembling chromosome no. 14 of North American species of its sister group taxon, supergenus Hydromantes. Acomparative analysis of C-band heterochromatin and silver-stained nucleolus organizer regions of Karsenia and other plethodontid genera reveals that chromosomal evolution may have featured chromosome 'repatterning' within the context of conserved chromosome number and shape in this clade. Genome size is correlated with geographic distribution in plethodontids and appears to have important phenotypic correlates as well. The genome size of Karsenia is relatively large, and resembles that of the geographically closest plethodontids from western North America, especially species of the genus Hydromantes. The biological significance of these cytogenetic characteristics of plethodontid salamanders is discussed within an evolutionary context. PMID- 18431683 TI - Elevated heavy metal concentrations in top soils of an Aegean island town (Greece): total and available forms, origin and distribution. AB - Elevated heavy metal concentrations in urban top soils are principal indicators of environmental pollution; however, relative data on the heavy metal status in soils of Greek island towns, that are regional administrative centers and popular tourist destinations, are missing. A survey was conducted to examine heavy metal concentrations in the urban soils of Ermoupolis, the capital of Syros island and of the prefecture of Cyclades complex in the Aegean Sea. Total (aqua-regia extracted) and available (DTPA extracted) concentrations of Cu, Pb, Zn, Ni, Cr, Sn and Fe were determined in top soil samples collected from green areas and open spaces of the town and in surface samples from inland reference soils of the island. Mean values for the aqua-regia extracted fraction of Cu, Pb and Zn were 117, 155 and 440 mg kg(-1) respectively, up to four times higher than the respective mean values of the reference soils. Enrichment factors (EFs) for these metals indicated high accumulation in the urban top soils and the available to total concentration ratio of Cu, Pb, Zn and Fe was higher for the urban compared to the reference soils, suggesting differences in metal sequestration, resulting in higher metal availability in the urban soils. GIS analysis was used to visualize the spatial distribution of EFs of the studied heavy metals. Factor Analysis and Cluster Analysis, applied to aqua-regia and DTPA data sets, adequately elucidated the origin of metals grouped under each factor or cluster. PMID- 18431684 TI - Analysis of airborne pollen grains in Bilecik, Turkey. AB - In this study, pollen grains in the atmosphere of Bilecik were studied for a continuous period of 2 years (2005 and 2006) by using Durham sampler. During this period, pollen grains belonging to 46 taxa were recorded, 26 of which belonged to arboreal plants and 20 to non-arboreal. Of total 14,269 pollen grains determined in Bilecik atmosphere, 6,675 were recorded in 2005 and 7,594 were in 2006. From these, 75.74% were arboreal, 21.80% were non-arboreal and 2.47% unidentifiable. Pinus sp., Poaceae, Cupressaceae, Platanus sp., Quercus sp., Salix sp., Ailanthus sp., Fagus sp., Urticaceae, Chenopodiaceae/Amaranthaceae were the main pollen producers in the atmosphere of Bilecik, respectively. Pollen concentrations reached their highest levels in May. Atmospheric pollen concentrations in February, March, September, October and November were less than those in other months. PMID- 18431685 TI - Biomechanics in young and adult italian standardbred trotter horses in real racing conditions. AB - This study was undertaken to characterize and establish the kinematic standards of Italian standardbred trotter horses in real racing conditions. Certain parameters, such as the angles of both front and hind fetlock joints, stride length, stride frequency and average speed in youngs and adults are presented, examinated and compared with statistical tests. The angles and stride length were obtained using ONTRACK software over frames extracted from two CANON MV630i digital videocameras. For the stride frequency, a third identical videocamera filmed the horses to count the number of beats per minute. The average speed was calculated by multiplying the stride frequency by the stride length and transforming the result into kilometers per hour. The data reveal that as speed increases, younger horses have more extended fetlock joints than adults. Contrary to this, at the maximum speed (hereafter referred to as sprint speed) the adults increase both their stride frequency and stride length, while young horses increase stride frequency and decrease stride length. We think that the knowledge of these parameters in competition breeds could have potential relevance as they may reveal early indicators of the development of proper and adequate characteristics in young horses of those breeds. PMID- 18431686 TI - N-glycan profiling in the study of human aging. AB - Most secreted proteins produced by the human body are modified by glycosylation. It is well known that the oligosaccharides (glycans) of glycoproteins are important for initiation of various cellular recognition signals that are essential for the maintenance of the ordered social life of each cell within a multi-cellular organism. The sugar chains can be altered by the physiological or pathophysiological condition of the cell. We describe a detailed protocol for the analysis of N-linked glycans in blood via DNA sequencing equipment-Fluorophore Assisted Carbohydrate Electrophoresis (DSA-FACE). The key features of this technique are its robustness, high throughput, high sensitivity and reliable quantification. Based on DSA-FACE technology, we previously reported that N glycan profiling of the human serum shows substantial changes with increasing age in three major N-glycan structures. We proposed that measurement of the N-glycan level changes could provide a surrogate marker for general health or for age related disease progression, and for monitoring the improvement of health after therapy. PMID- 18431687 TI - Late contrast enhancement by CMR: more than scar? PMID- 18431688 TI - The complete mitochondrial genome structure of snow leopard Panthera uncia. AB - The complete mitochondrial genome (mtDNA) of snow leopard Panthera uncia was obtained by using the polymerase chain reaction (PCR) technique based on the PCR fragments of 30 primers we designed. The entire mtDNA sequence was 16 773 base pairs (bp) in length, and the base composition was: A-5,357 bp (31.9%); C-4,444 bp (26.5%); G-2,428 bp (14.5%); T-4,544 bp (27.1%). The structural characteristics [0] of the P. uncia mitochondrial genome were highly similar to these of Felis catus, Acinonyx jubatus, Neofelis nebulosa and other mammals. However, we found several distinctive features of the mitochondrial genome of Panthera unica. First, the termination codon of COIII was TAA, which differed from those of F. catus, A. jubatus and N. nebulosa. Second, tRNA(Ser) ((AGY)), which lacked the ''DHU'' arm, could not be folded into the typical cloverleaf shaped structure. Third, in the control region, a long repetitive sequence in RS 2 (32 bp) region was found with 2 repeats while one short repetitive segment (9 bp) was found with 15 repeats in the RS-3 region. We performed phylogenetic analysis based on a 3 816 bp concatenated sequence of 12S rRNA, 16S rRNA, ND2, ND4, ND5, Cyt b and ATP8 for P. uncia and other related species, the result indicated that P. uncia and P. leo were the sister species, which was different from the previous findings. PMID- 18431689 TI - Quantitative second harmonic generation imaging of cartilage damage. AB - Cartilage damage was studied using non-invasive multiphoton-excited autofluorescence and quantitative second harmonic generation (SHG) microscopy. Two cryopreservation techniques based upon freezing and vitrification methods, respectively, were employed to determine whether or not the collagen fiber structure of full thickness porcine articular cartilage was affected by cryopreservation and whether the level of collagen damage could be determined quantitatively in non-processed (non-fixed, non-sliced, non-stained) tissues. Multiphoton-induced autofluorescence imaging revealed the presence of chondrocytes, as well as collagenous structures in all fresh, vitrified and frozen cryopreserved cartilage samples. SHG imaging of the frozen cryopreserved specimens showed a dramatic loss of mean gray value intensities when compared to both fresh and vitrified tissues (P<0.05), indicating structural changes of the extracellular matrix, in particular the deformation and destruction of the collagen fibers in the analyzed articular cartilage. A 0.9974 correlation coefficient was observed between the metabolic cell activity assessed by the alamarBlue technique, and retention of collagen structure between the three experimental groups. These studies suggest that multiphoton-induced autofluorescence imaging combined with quantitative SHG signal profiling may prove to be useful tools for the investigation of extracellular matrix changes in preserved cartilage, giving insights on the structural quality prior to implantation. PMID- 18431690 TI - Effect of low dose and moderate dose gamma irradiation on the mechanical properties of bone and soft tissue allografts. AB - The increased use of allograft tissue for musculoskeletal repair has brought more focus to the safety of allogenic tissue and the efficacy of various sterilization techniques. Gamma irradiation is an effective method for providing terminal sterilization to biological tissue, but it is also reported to have deleterious effects on tissue mechanics in a dose-dependent manner. At irradiation ranges up to 25 kGy, a clear relationship between mechanical strength and dose has yet to be established. The aim of this study was to investigate the mechanical properties of bone and soft tissue allografts, irradiated on dry ice at a low absorbed dose (18.3-21.8 kGy) and a moderate absorbed dose (24.0-28.5 kGy), using conventional compressive and tensile testing, respectively. Bone grafts consisted of Cloward dowels and iliac crest wedges, while soft tissue grafts consisted of patellar tendons, anterior tibialis tendons, semitendinosus tendons, and fascia lata. There were no statistical differences in mechanical strength or modulus of elasticity for any graft irradiated at a low absorbed dose, compared to control groups. Also, bone allografts and two soft tissue allografts (anterior tibialis and semitendinosus tendon) that were irradiated at a moderate dose demonstrated similar strength and modulus of elasticity values to control groups. The results of this study support the use of low dose and moderate dose gamma irradiation of bone grafts. For soft tissue grafts, the results support the use of low dose irradiation. PMID- 18431691 TI - A mathematical approach predicting the number of events in different tumors. AB - Supported by different investigations, multi-step models for tumorigenesis have been proposed for epithelial tumors. The age specific incidence of some cancers shows an exponential rise with increasing patient age. Yet, the onset and the slope of incidence curves varies between tumor types. One simple explanation for this disparity is that the number of mutations required for transformation differs in various tissues. We used a homogeneous Poisson process to estimate the number of events (N) and the intensity or event rate (lambda) that might be needed for cancer development in various tissues (colon, prostate, oralpharynx, larynx). Estimations were performed, including 95% confidence intervals, for the male and female population. The expected number of events needed was higher in adenocarcinomas (colorectal carcinoma: N approximately 10 for females and N approximately 11.0 for males; prostatic cancer: N approximately 23) than in squamous cell carcinomas (oropharynx: N approximately 5-6 for females and N approximately 6 for males; larynx: N approximately 7 for females and N approximately 8 males). Still, alternative models fixing N to values within the 95% confidence intervals determined, showed good coincidence with epidemiological data. Although the herein applied mathematical model neglects several biologic conditions, especially a presumed acceleration of mutation rates after tumor initiation it offers a plausible theory for the given epidemiologic data and matches with molecular biologic findings in the investigated cancers. PMID- 18431692 TI - Microscopic peliosis of pancreatic islets associated with thrombotic thrombocytopenic purpura. AB - Peliosis is a rarely seen histological finding with unexplained fully etiology and pathogenesis. It is presented as cyst-like blood filled cavities. The presence of peliosis in the endocrine part of the pancreas is extremely rarely reported microscopic phenomenon. The authors provide histological, histochemical, immunohistochemical and ultrastructural investigation of microscopic peliosis in the pancreas from an autopsy case with thrombotic trombocytopenic purpura. The findings give ideas for a wide range of pathophysiological and morphogenetic comments of such an unusual morphologic presentation. PMID- 18431693 TI - Ryanodine receptor expression correlates with tumor grade in breast cancer. AB - Ryanodine receptors (RyRs) have been previously implicated in the proliferation of human T-lymphocytes and melanocytes as well as in the motility of astrocytes. We have examined RyR expression in 57 ductal, human breast cancer specimens, by immunohistochemistry. Moderate to high RyR immunostaining was found in 47 (82%) of the specimens. There was a direct correlation between RyR levels and tumor grade (r = 0.48, p = 0.0002). We have also examined the effect of the RyR agonist 4-chloro-m-cresol on the in-vitro growth of two human breast cancer cell lines, MCF-7 and MDA-MB-231. Treatment with 4-chloro-m-cresol inhibited the growth of both breast cancer cell lines, in a dose-dependent manner, with half-maximal inhibition observed at 30 to 50 microg/mL (210-351 microM). Our data suggest that RyR could serve as prognostic indicator and/or as a target for breast cancer treatment. PMID- 18431694 TI - Nodular lymphocyte predominant Hodgkin lymphoma (NLPHL)-clinicopathological features based on the data of two Hungarian lymphoma centres. AB - Clinicopathological features of nodular lymphocyte predominant Hodgkin lymphoma (NLPHL) differ from those of the classical Hodgkin lymphoma (cHL). Our aim was to examine clinical presentation, therapeutic and survival results of NLPHL patients in Hungary based on the data of two centres, and incidentally we analyzed the clinicopathological characteristics and differential diagnostic difficulties of this rare entity. We analyzed the clinical features, treatment and survival data of 536 Hodgkin lymphoma patients who had been diagnosed and primarily treated in our institutes between 1995 and 2004. Mean follow-up time was 82.7 (3-144) months of the total 536 HL patients. Sixteen (3%) of the patients were diagnosed with NLPHL, 93% of them presented with early-stage disease. None of the patients showed extranodal or splenic involvement or bulky disease. One patient received chemotherapy alone, six received only involved field radiotherapy while six underwent combined modality treatment. We applied watch and wait strategy in three cases. Overall response rate was 100% (93.75% complete). Two NLPHL cases transformed to non-Hodgkin's lymphoma. In contrast to the classical HL, the 10 year prognosticated overall survival rate was 100 vs. 82%, the event free survival was: 75% vs. 70%. In NLPHL group there were no late or multiple relapses and none of them died. CONCLUSIONS: NLPHL is a rare disease, thus these are limited experiences with its diagnosis and treatment. Since the disease has an excellent outcome, it is very important to prefer less toxic or local therapies to reach long term survival similar to that of the normal population. PMID- 18431695 TI - Orbital tumour as initial manifestation of acute myeloid leukemia: granulocytic sarcoma: case report. AB - We report orbital involvement as an initial manifestation of acute myeloid leukemia in a 57-year-old woman. The patient presented with painful proptosis and limited ocular motility. Orbital computed tomography revealed bilateral homogeneous masses. Orbital biopsy was performed on the right side; and histopathology disclosed a myelocytic tumour. Despite treatment using irradiation and chemotherapy, the patient died eleven months after presentation. There appear to be only a few previous reports of acute myeloid leukemia cases presenting with orbital involvement, and most cases occurred in children. This very rare condition has a poor survival prognosis, even with radiation treatment and chemotherapy. PMID- 18431696 TI - The rise of fluoroquinolone resistant Neisseria gonorrhoeae. Implications for treatment guidelines. PMID- 18431697 TI - Biomarkers: past, present, and future. AB - In recent decades biomarkers have become accepted tools in clinical practice [1]. Although there is no widely accepted definition of what constitutes a biomarker, for the context of this review we consider a biomarker to be a protein or other macromolecule that is associated with a biological process or regulatory mechanism. Hence measurement of this biomarker in blood, for example, might provide quantitative information that could be clinically helpful regarding this biological process or regulatory mechanism. In this paper we review recent advances with the use of biomarkers in three major clinical areas: diagnosis of myocardial infarction, diagnosis and management of heart failure, and diagnosis and management of inflammatory conditions in general and systemic infections in particular. Although these may look like unrelated medical challenges, recent clinical research in these areas by our groups and others has opened up opportunities and challenges that seem fundamental for biomarkers in general. PMID- 18431698 TI - Non-invasive diagnosis of coronary artery disease using cardiogoniometry performed at rest. AB - PRINCIPLES: Cardiogoniometry is a non-invasive technique for quantitative three dimensional vectorial analysis of myocardial depolarization and repolarization. We describe a method of surface electrophysiological cardiac assessment using cardiogoniometry performed at rest to detect variables helpful in identifying coronary artery disease. METHODS: Cardiogoniometry was performed in 793 patients prior to diagnostic coronary angiography. Using 13 variables in men and 10 in women, values from 461 patients were retrospectively analyzed to obtain a diagnostic score that would identify patients having coronary artery disease. This score was then prospectively validated on 332 patients. RESULTS: Cardiogoniometry showed a prospective diagnostic sensitivity of 64%, and a specificity of 82%. ECG diagnostic sensitivity was significantly lower, with 53% and a similar specificity of 75%. CONCLUSIONS: Cardiogoniometry is a new, noninvasive, quantitative electrodiagnostic technique which is helpful in identifying patients with coronary artery disease. It can easily be performed at rest and delivers an accurate, automated diagnostic score. PMID- 18431699 TI - Assisted suicide in an acute care hospital: 18 months' experience. AB - QUESTION UNDER STUDY: In 2006 the University Hospital of Lausanne (CHUV) introduced an institutional directive specifying the conditions for assisted suicide, in accordance with professional guidelines and the recommendation of the Swiss National Advisory Commission on Biomedical Ethics that every acute care hospital take up a position on this subject. METHODS: 18-months follow-up analysis of patient requests and application of the directive by hospital staff. RESULTS: Of the 54,000 patients hospitalised between January 1, 2006, and June 30, 2007, six requests were recorded, all within the first 7 months after introduction of the directive and in the context of severe and life-threatening diseases. However, only one of the six patients, living in a nursing home belonging to the hospital, died by assisted suicide. Two patients died from their diseases, one during the assessment procedure and the other shortly after. One patient withdrew his request after pain control, returned home and died several weeks later. Another patient, although she was severely ill and died several months later, was denied the procedure because her condition was improving. Only one patient was declared incompetent and his request refused. The time distribution of requests seems to be associated with initial media coverage of the assisted-suicide directive's introduction. Only minor amendments to the directive were needed. CONCLUSIONS: The recommendations of the Swiss National Advisory Commission on Biomedical Ethics are applicable in an acute care hospital. PMID- 18431700 TI - A new sexually transmitted infection (STI) in Geneva? Ciprofloxacin-resistant Neisseria gonorrhoeae, 2002-2005. AB - PRINCIPLES: Neisseria gonorrhoeae (NG) resistant to ciprofloxacin (CR) was documented for the first time in Geneva in 2002 and increased from 7% that year to 47% in 2005. We describe NG cases during this period and compare characteristics of CR and ciprofloxacin susceptible (CS) cases. METHOD: Geneva microbiological laboratories identified NG cases. Antimicrobial susceptibility testing (AST) was performed on a sample of reported cases. The attending physicians completed questionnaires on demographic and epidemiological characteristics. Risk exposures were assessed by comparing CR and CS cases using logistic regression. RESULTS: 238 NG cases were reported. Of 91 on which AST was performed, 23 (25%) were CR; 91% of these were male vs 87% of CS patients. Men having sex with men (MSM) represented 38% of CR cases compared with 31% of CS cases (p>0.05). Among CR cases 65% were Swiss compared with 56% of CS cases. Median age was 35 years for both. Casual sexual contacts were reported for 88% of CR cases and 72% of CS cases (p>0.05). The difference between CR and CS cases in terms of sexual activity outside Switzerland (50% and 19% respectively) remained significant after adjusting for sexual preference and nationality (OR: 7.0, CI 95: 1.99-24.6). CONCLUSION: Although CR infection was more common among Swiss MSM, only sexual activity outside Switzerland was independently associated with CR. Physicians should request AST before treatment and reconsider first-line use of ciprofloxacin. Surveillance of gonococcal antimicrobial resistance is essential in monitoring epidemiologic trends and updating recommendations on first-line treatment. PMID- 18431701 TI - [Time-adaptive mode, a new ventilation form for the treatment of respiratory insufficiency--a self-learning system]. AB - Hypercapnic respiratory failure is usually caused by an overload of the respiratory muscles (respiratory pump). After treatment of the underlying disease, mechanical ventilation will achieve optimal treatment success and higher degrees of respiratory muscle unloading will improve the outcome in terms of lower PaCO (2) levels and improved exercise performance. Routinely assisted modes are being used for ventilation, where the patient has to trigger the ventilator with his effort. Controlled ventilation is usually applied in sedated patients lacking spontaneous breathing efforts that are necessary to trigger the ventilator. Controlled ventilation, however, is feasible in awake patients but requires operator expertise. In this process, the respiratory pattern of the ventilator has to be adapted to the patient's own respiratory pattern. Changing conditions require a re-adaptation of parameters. In order to automatise this complex and time-consuming operation, a time-adaptive mode (TA-mode) has been developed. This programmed mode incorporates a self-learning algorithm, primarily detecting the patient's respiratory pattern. The software then calculates a matching flow profile using a motion equation that gives consideration to resistance and compliance. The operator has to pre-select allowed ranges of parameters (especially in- and expiratory pressures, IPAP and EPAP). After detection of a stable respiratory pattern (usually after 10 - 20 breaths), the ventilator will slowly increase the calculated flow profile and achieve controlled ventilation without irritating respiratory centres of the brain. Respiratory drive will cease usually within three to five minutes. Restart of the respiratory drive, for example, after coughing or during REM sleep with an altered respiratory pattern will be detected as ventilator fighting and the programme will return to the analysis algorithm again. After the respiratory pattern has become stable, the ventilator will take over ventilation again. The new mode has been validated in an accreditation study. For this purpose we selected 21 patients with stable hypercapnic respiratory failure, most of whom (20) had previously been ventilated with a controlled T-mode and only one patient had previously been ventilated with an assisted mode and adapted them to the new ventilator under polygraphic surveillance. Each time seven patients were adapted to a T-, ST- and TA-mode, respectively. Two patients, however, could not be adapted to ST-mode ventilation and were switched to TA-mode. PCO (2) values before and after ventilation were not significantly different between modes. Patient satisfaction was rated very good in 34 %, good in 45 % and non-gratifying in 21 % of cases ventilated with TA-mode. Consideration has to be given to the fact that patients previously had been receiving optimal ventilator treatment. The TA-mode is a self-learning system, capable of copying the patients own breathing pattern while awake, in order to achieve complete unloading of the respiratory muscles through controlled ventilation during a circumscribed period. PMID- 18431702 TI - [Validity of information relevant to research in routine medication claims data from 2000 to 2006]. AB - BACKGROUND: Claims data of German statutory health insurance companies have been used increasingly over the last years for various research purposes. However, little is known about the validity of these data. Aim of this study was to analyse whether information relevant for research purposes is accurately recorded in computerized out-patient medication claims data. METHODS: We used data of the Gmunder ErsatzKasse (GEK) over the years 2000 - 2006. A random sample of 300 prescriptions annually was drawn from a total of 6.3 to 7.5 million prescriptions. Data on the scanned original prescriptions ("gold standard") were validated against that information in claims data. RESULTS: Over the years 2000 - 2006 an increasing proportion of dates of writing was recorded accurately (77.3 % to 93.0 %; p for trend < 0.0001). Concomitantly the proportion of handwritten prescriptions declined and that of printed ones increased. Dates of dispensing were barely correct until 2003. Thereafter, the agreement ranged from 65.3 % in 2004 to 86.1 % in 2006 (p for trend < 0.0001). There were significant differences between pharmacy data processing centres (Apothekenrechenzentren). The accuracy of physician and pharmacy identifiers (97,7%, 99,75% and 100.0 %, respectively) was nearly constant over these 7 years. CONCLUSION: The accuracy of variables analysed in this study is very good for those which are immediately related to the flow of money in German out-patient medication claims data. Recording of dates of prescription and dispensing are fairly good quality. PMID- 18431703 TI - [Portal vein hypertension during azathioprine therapy in patients with Crohn's disease--a frequent phenomenon?]. AB - HISTORY AND FINDINGS ON ADMISSION: A 45-year-old man with 18 years history of Crohn's disease who was on treatment with azathioprine (AZA) for the past 48 months was admitted because of haematemesis and melaena. INVESTIGATIONS AND DIAGNOSIS: Tests showed a slight elevation of bilirubin and a low platelet count. Endoscopy revealed esophageal and gastric fundus varices. Magnetic resonance imaging (MRI) revealed severe portal vein hypertension with a spleen-kidney convolute of varices. Liver biopsy showed hyperplasia of hepatocytes and a reticular fibrosis, consistent with the diagnosis of nodular regenerative hyperplasia (NRH). TREATMENT AND COURSE: AZA treatment was stopped. The fundal varices were treated with endoscopic histoacryl injection. The patient is at present in good health and is followed up in our outpatient department. CONCLUSION: AZA as a widely used immunosuppressive drug has side effects in about 5 - 10 % of the cases. The risk of hepatotoxicity in patients treated with AZA is often underestimated. Therefore, physicians treating patients with inflammatory bowel disease have to be precocious regarding the increase of hepatic enzymes and pathologic signs in liver imaging in relation of AZA treatment. The latter might refer to NRH which would lead to severe portal vein hypertension. PMID- 18431705 TI - [Pseudoxanthoma elasticum with "angioid streaks" (Gronblad-Strandberg syndrome)]. PMID- 18431704 TI - [Molecular pathogenesis of proteinuria]. PMID- 18431706 TI - [Eating disorders--diagnosis and treatment]. AB - Anorexia nervosa and Bulimia nervosa are the main types of eating disorders described in the International Classification of Diseases ICD-10. The main features are eating patterns such as refusal to eat enough food or loss of control, followed by counter-regulatory measures. In addition, preoccupation with body shape and weight and with food is an important feature of eating disorders. Severe medical conditions may occur as a result to starvation, malnutrition and purging. Binge eating disorder has been included as an additional variant of disturbed eating in the American classification system of mental disorders (DSM IV). The main characteristic of the binge eating disorder is binge eating, but without counter-regulatory measures. Patients with anorexia nervosa are foremost underweight (BMI < 17.5 kg/m2), those with a bulimia nervosa are usually in the normal weight range. On the other hand, patients with the binge eating disorder are overweight or obese. Etiological models are multifaceted and include predisposing and sustaining factors as well as triggers for the onset of the disorder. The course is variable and marked by changes between remission and clinically relevant symptoms. Psychotherapy is the treatment of choice. PMID- 18431707 TI - [Eosinophilic esophagitis: new standards in diagnosis and therapy of chronic retrosternal pain]. AB - Eosinophilic esophagitis is a chronic allergic inflammatory condition of the esophagus, which most often results in dysphagia, bolus impaction, heartburn or chest pain. Of particular importance is the differentiation from other inflammatory diseases of the esophagus, especially gastro-esophageal reflux disease. Response to treatment with proton pump inhibitors may help to distinguish between the different entities. The most important element in the diagnosis of eosinophilic esophagitis is to know its macro- and micromorphological characteristics. Biopsies from the proximal to the distal esophagus demonstrating > 15-20 eosinophilic granulocytes per high powered field favor the diagnosis. A multimodal therapeutic concept consists of the avoidance of specific allergens, topical or systemic glucocorticoids, oral antihistamines, leukotriene antagonists and, in cases of co-existing gastro-esophageal reflux disease, oral intake of proton pump inhibitors. PMID- 18431708 TI - [The Gottingen palliative emergency card: improvement of emergency medical care for ambulatory palliative care patients. The "yellow card for rescue services"]. AB - Currently more and more patients with cancer will be treated at home, especially at the final stage of their disease. Relatives and nursing services will therefore be confronted with acute emergency situations concerning these patients. The handling of these acute situations may be beyond the relatives' coping capacity. For that reason the pre-hospital emergency system (2.5% of all emergency calls) will be confronted with very specific needs of these patients. Emergency situations of palliative patients at home are more predictable than other circumstances. It is thus possible to achieve a practicable preparation for such emergency situations. An individual "emergency plan" should be created for these cases.. The plan may be processed in an emergency situation. It may then be possible to avoid emergency calls and make it possible for the patient to remain at home. This promotes the patient's quality of life and his/her predominant desire to stay at home after the emergency. An advance directive should be made which records the patient's wishes. Because of the complexity of an advance directive it is essential to indicated the patient's wishes in a short and quickly read form in case an emergency arises. This is one of the reasons for producing a short version like the "Gottingen palliative emergency card". In acute situations it is then possible quickly to make known the patient's wishes The patient may be given the possibility of remaining at home after an emergency situation has been dealt with successfully. PMID- 18431710 TI - [The voice]. PMID- 18431709 TI - [Sialendoscopy: diagnostic possibilities and therapeutic options]. AB - With the improved availability of sialendoscopy (SE) during the last decade the therapeutical concept for sialoliths has remarkably changed. Due to the introduction of SE the diagnostic opportunities could be extended. Before introduction of SE, the diagnostic was focused only on the existence and size of sialoliths. Now, questions like consistence of sialoliths and their mobility in the duct system, stenoses with sialolithiasis-like symptoms as well as the evaluation of the remaining secretory capacity of the involved gland became relevant for the therapeutic decision. Furthermore there are an increasing number of therapeutic opportunities available. For some of these techniques long term experiences are missing so far. On the basis of the review of the current literature and a retrospective analysis of 256 own patients the different techniques and their indication are described. The combination of different therapeutical options of the SE and the still established extracorporal shockwave lithotripsy (ESWL) results in an important reduction of glandular resection in patients with sialolithiasis. Thus, the rate of glandular resection for the glandular submandibular can be decreased to 5 % and for the glandular parotis to 1 %. In summary, the most effective way is a stepwise approach based on the diagnostic tools of SE and including the laser-lithotripsy and ESWL in the treatment of sialolithiasis. PMID- 18431711 TI - [From the expert's office: could a medical expert be hold liable for the results of his expertise?]. PMID- 18431712 TI - [Computer-assisted intraoperative navigation at the anterior skull base]. AB - Intraoperative 3D-navigation at the anterior skull base has become a very valuable tool in the last years. For a successful use the clinical pathways need a slight adaptation only to provide the radiologic imagery to the system. Established algorithms and standardized protocols have proven 3D-navigation systems as a valuable clinical tool, when used in conjunction with appropriate intraoperative quality assurance. Ease-of-use and reliable intraoperative quality assurance is an active area of research that, combined with adequate strategies for referencing the patient to preoperative high-resolution radiologic data, will make 3D-navigation at the lateral skull base a successful clinical tool as well. PMID- 18431713 TI - [Update on auditory synaptopathy/auditory neuropathy (AS/AN)]. PMID- 18431714 TI - [Mechanisms of nicotine dependence]. AB - About 30 % of the population in Western societies smoke. Most smokers do so due to nicotine dependence. In concert with ongoing education about the detrimental consequences of tobacco abuse and further restriction of public smoking, further scientific effort is needed to investigate the mechanisms of nicotine dependence, in order to develop more effective treatments and smoking cessation programmes. This review summarises our current knowledge of the mechanisms of nicotine dependence, focussing mainly on the cellular effects of nicotine and the effects on three neurophysiological systems that contribute to nicotine dependence: a) reward system, b) cognition/attentional networks and c) stress response system. The reward system that is connected with the mood regulatory system is activated by nicotine and other addictive substances. Furthermore, nicotine modulates cognitive networks involved in attention and learning/memory. Most data point to positive effects of acute nicotine administration on these networks. Finally nicotine influences the stress response system, however, the effects depend on the stage of nicotine addiction. Nicotinic modulation of these networks by means of smoking may reflect an attempt to self-medicate clinical or subclinical symptoms in the areas of mood regulation/depression, attention and learning/memory and stress coping, at least in a subset of smokers. PMID- 18431716 TI - [Non-surgical treatment of mallet finger fractures involving more than one third of the joint surface: 10 cases]. AB - Operative stabilisation of mallet finger fractures is generally recommended in cases with more than one-third of the articular surface involved. Results of surgical stabilisation are often unsatisfying regarding anatomic reconstruction of the joint surface and mobility of the DIP joint. In a retrospective study we present the results of conservative treatment of 10 consecutive patients with mallet finger fractures involving one-third to two-thirds of the joint surface. Treatment consisted in splinting with a dorsal aluminium splint for 4 to 6 weeks, nightsplinting continued for a total of 8 weeks. Functional results at 9 months follow-up are very good with an extensor lag of maximally 5 degrees in 5 cases, a reduced flexion of maximally 10 degrees in 2 cases. All patients are pain-free. Radiological results show a very good remodelling of the DIP joint surface with anatomic joint congruency even in cases with initially up to 3 mm fragment displacement and 1 mm fragment rotation. There was no secondary palmar subluxation. Our findings support the conclusion of Wehbe and Schneider and Tuttle et al. that most mallet finger fractures can be treated conservatively, regardless the size and amount of displacement of the bone fragment. In conclusion, we consider that operative stabilisation has to be discussed only in the presence of palmar subluxation. PMID- 18431717 TI - [Recurrence of solitary enchondroma at the hand after operative treatment]. AB - PURPOSE: Solitary enchondroma is the most common bone tumour at the hand. Nevertheless there are only a few studies referring to a recurrence of this lesion after operative treatment. PATIENTS AND METHOD: 21 patients (17 women/4 men) were examined retrospectively 2 to 18 (mean: 9) years after operative treatment. Clinical and radiological examinations were performed. 3 to 4 years later 9 of these patients were re-examined. 1 patient was first seen at the time of the second examination. Recurrence was the endpoint of the study. RESULTS: At the first examination 11 patients had normal cancellous bone structure 4 to 18 (mean: 8) years after operative treatment. 7 patients had remaining bone defects and their follow-up was 2 to 15 (mean: 8) years. Recurrence was found in 3 patients 11 to 17 years postoperatively. Through the re-examination of 9 patients 2 further recurrences were found 4 to 6 or, respectively, 11 to 14 years postoperatively arising from remaining defects. Another remaining defect became normal cancellous bone structure 2 to 5 years postoperatively. Over all 6 recurrences were found in 22 patients. All patients were free of clinical symptoms. CONCLUSION: Recurrence may occur even more than 10 years postoperatively. Therefore we recommend periodical radiological re-examination for recurrence before weakness of bone can lead to a pathological fracture. Re examination intervals of 2 to 5 years would probably be adequate. PMID- 18431718 TI - Overexpression of TRPC3 reduces the content of intracellular calcium stores in HEK-293 cells. AB - The mammalian canonical transient receptor channels (TRPCs) are considered to be candidates for store-operated calcium channels (SOCCs). Many studies have addressed how TRPC3 channels are affected by depletion of intracellular calcium stores. Conflicting results have been shown for TRPC3 regarding its function, and this has been linked to its level of expression in various systems. In the present study, we have investigated how overexpression of TRPC3 interferes with the regulation of intracellular calcium stores. We demonstrate that overexpression of TRPC3 reduces the mobilization of calcium in response to stimulation of the cells with thapsigargin (TG) or the G-protein coupled receptor agonist sphingosine-1-phosphate (S1P). Our results indicate that this is the result of the expression of TRPC3 channels in the endoplasmic reticulum (ER), thus depleting ER calcium stores. OAG evoked calcium entry in cells overexpressing TRPC3, indicating that functional TRPC3 channels were also expressed in the plasma membrane. Taken together, our results show that overexpression of the putative SOCC, TRPC3, actually reduces the calcium content of intracellular stores, but does not enhance agonist-evoked or store-dependent calcium entry. Our results may, in part, explain the conflicting results obtained in previous studies on the actions of TRPC3 channels. PMID- 18431719 TI - A sensitive and selective immuno-nanogold resonance-scattering spectral method for the determination of trace penicillin G. AB - A sensitive and selective immuno-nanogold resonance scattering spectral assay was developed for the determination of trace hapten penicillin G, based on the resonance scattering (RS) effect of the nanogold at 560 nm, and the nanogold labelled immunoreaction took place in pH 5.4 phosphate citric acid buffer solutions and in the presence of polythylene glycol (PEG). The nanogold-labelled immunocomplex formed more and more with addition of penicillin G. The enhanced RS intensity at 560 nm Delta I(RS) was linear to the penicillin G concentration in the range 7.5-1700 ng/mL, with a detection limit of 0.78 ng/mL. The results indicate that the immunonanogold-labelled RS spectral assay has a high specificity and sensitivity for quantitative determination of penicillin G in raw milk samples. PMID- 18431720 TI - Diagnostic utility of phosphatase and tensin homolog, beta-catenin, and p53 for endometrial carcinoma by thin-layer endometrial preparations. AB - BACKGROUND: For the current report, the authors examined the characteristic features of morphology and molecular biology of phosphatase and tensin homolog (PTEN), beta-catenin, and p53 immunocytochemistry in endometrial carcinoma by using thin-layer cytologic preparations. METHODS: During a 6-month period, 120 endometrial samples were collected directly by using the Uterobrush, and thin layer specimens were prepared. Immunocytochemical expression levels of PTEN, beta catenin, and p53 were investigated by using 40 specimens of endometrial carcinoma (EC), and 30 specimens each of proliferative endometrium, secretory endometrium, and atrophic endometrium. RESULTS: For PTEN immunoreactivity, the a cutoff value of 50% PTEN expression appeared to be useful for the correct diagnosis of EC in endometrial cytology. For beta-catenin immunoreactivity, an increase in cytoplasmic and nuclear beta-catenin expression and a loss of beta-catenin expression appeared to be useful for the correct diagnosis of EC in endometrial cytology and may aid in the stratification of EC into low grade and high grade EC. For p53 immunoreactivity, the application of a cutoff score >or=4 for nuclear p53 expression appeared to be useful for the diagnosis of high-grade EC in endometrial cytology. CONCLUSIONS: Immunocytochemical findings from a combination of PTEN, beta-catenin, and p53, in addition to cytomorphologic features, appeared to be useful for the more accurate diagnosis of EC in endometrial cytology. PMID- 18431721 TI - Fabrication of suspended silicon nanowire arrays. AB - A method to fabricate suspended silicon nanowires that are applicable to electronic and electromechanical nanowire devices is reported. The method allows for the wafer-level production of suspended silicon nanowires using anisotropic etching and thermal oxidation of single-crystal silicon. The deviation in width of the silicon nanowire bridges produced using the proposed method is evaluated. The NW field-effect transistor (FET) properties of the device obtained by transferring suspended nanowires are shown to be practical for useful functions. PMID- 18431722 TI - Fabrication of colloidal crystal beads by a drop-breaking technique and their application as bioassays. PMID- 18431724 TI - Total synthesis of (+)-fawcettidine. PMID- 18431723 TI - Sc3N@C80-ferrocene electron-donor/acceptor conjugates as promising materials for photovoltaic applications. PMID- 18431725 TI - Wavelength-dependent photochromic inorganic-organic hybrid based on a 3D iodoplumbate open-framework material. PMID- 18431726 TI - Synthesis and screening of an oroidin library against Pseudomonas aeruginosa biofilms. AB - A 50-compound library based on the marine natural product oroidin was synthesized and assayed for anti-biofilm activity against PAO1 and PA14, two strains of the medically relevant gamma-proteobacterium Pseudomonas aeruginosa. Through structure-activity relationship (SAR) analysis of analogues based on the oroidin template, several conclusions can be drawn as to what structural properties of the synthetic derivatives are necessary to elicit a biological response. Notably, the most active analogues identified were those that contained a 2-aminoimidazole (2-AI) motif and a dibrominated pyrrolecarboxamide subunit. Here we disclose the synthesis and subsequently determined biological activity of this unique class of compounds as inhibitors of biofilm formation that have no direct antibiotic effect. PMID- 18431727 TI - Two diammonium copper azides with similar layerlike magnetic substructures made of chains of serially connected Cu6 rings show cation-modulated magnetism. AB - We present here the first examples of Cu-azide compounds synthesized by using protonated diamine ions as cationic templates: (dmenH(2))[Cu(6)(N(3))(14)] and (trimenH(2))[Cu(6)(N(3))(14)](dmenH(2)(2+): N,N'-dimethylethylenediammonium; trimenH(2)(2+): N,N,N'-trimethylethylenediammonium). Both compounds possess a similar, rarely observed anionic Cu-azide layer, which consists of [Cu(6)(N(3))(14)(2-)](n) anionic chains linked by asymmetric end-to-end azido bridges. The chain, in turn, is made up of elongated Cu(6) rings, with double and single end-on azido linkages between the square-planar Cu(2+) sites within the ring and double end-on azido bridges serially connecting the rings. The molecular geometry results in ferromagnetic interactions within the Cu-azide layer in both compounds. The interlayer separations are determined by the cations, with the shortest interlayer CuCu separations being 8.016 A for the dmenH(2)(2+) compound and 9.106 A for the trimenH(2)(2+) compound. These different interlayer separations tune the magnetic properties of the two materials. The dmenH(2)(2+) compound displays long-range antiferromagnetic ordering at low temperature and short-range ferromagnetic interaction at high temperature, while only short-range ferromagnetism was observed in the trimenH(2)(2+) compound at 2-300 K. PMID- 18431728 TI - Catalytic amide-mediated methyl transfer from silanes to alkenes in Fujiwara Moritani oxidative coupling. PMID- 18431729 TI - Theoretical trends of diffusion and reaction into tubular nano- and mesoporous structures: general physicochemical and physicomathematical modeling. AB - A general and adaptable physicochemical model is presented to evaluate the mass transport within nanopores of mesoporous particles when the mass transport is coupled to heterogeneous kinetics occurring at active sites located onto the nanopore walls surface. The model framework encompasses almost all situations of practical interest in solutions and may be used for characterizing the kinetic rates and constants controlling the system under different sets of experimental conditions. Furthermore, it allows the delineation of simple effective parameters, which should be most useful for optimizing a given material in view of specific applications. For the sake of clarification the simplified model is presented and its results discussed by specializing it for cases where the reactions involve a simple adsorption of a target species on the nanopore immobilized sites as observed for inorganic sponges used in water decontamination. Yet it may easily be extended further to encompass a wider variety of situations where the sites immobilized onto the nanopore walls perform chemical or biochemical transformations as occur in supported catalysis in liquid solution. PMID- 18431730 TI - Oxoanion binding by guanidiniocarbonylpyrrole cations in water: a combined DFT and MD investigation. AB - Structures and properties of nonbonding interactions involving guanidinium functionalized hosts and carboxylate substrates were investigated by a combination of ab initio and molecular dynamics approaches. The systems under study are on one hand intended to be a model of the arginine-anion bond, so often observed in proteins and nucleic acids, and on the other to provide an opportunity to investigate the influence of molecular structure on the formation of supramolecular complexes in detail. Use of DFT calculations, including extended basis sets and implicit water treatment, allowed us to determine minimum energy structures and binding enthalpies that compared well with experimental data. Intermolecular forces were found to be mostly due to electrostatic interactions through three hydrogen bonds, one of which is bifurcate, and are sufficiently strong to induce a conformational change in the ligand consisting of a rotation of about 180 degrees around the guanidiniocarbonylpyrrole axis. Free binding energies of the complexes were evaluated through MD simulations performed in the presence of explicit water molecules by use of the molecular mechanics Poisson-Boltzmann solvent accessible surface area (MM-PBSA) and linear interaction energy (LIE) approaches. LIE energies were in quantitative agreement with experimental data. A detailed analysis of the MD simulations revealed that the complexes cannot be described in terms of a single binding structure, but that they are characterized by a significant internal mobility responsible for several low-energy metastable structures. PMID- 18431731 TI - Dimeric capsules with a nanoscale cavity for [60]fullerene encapsulation. AB - The acid-assisted and guest-induced formation of superstructures was achieved by the addition of haloacetic acids to a toluene solution of the resorcin[4]arene derivatives 1 and [60]fullerenes. The formation of dimeric superstructures that encapsulated a nanosized guest molecule was observed when appropriate acids, such as haloacetic acids, and suitable guest molecules, such as [60]fullerenes, were co-added to a toluene solution of cavitand 1 that has four pyridine units, whereas a complicated equilibrium between several species was detected without [60]fullerenes, and the formation of discrete superstructures was not monitored in the absence of haloacetic acids. The spectroscopic data indicate that the formed [60]fullerene-encapsulated complexes have the structure of 2. These complexes are self-assembled through pyridinium-anion-pyridinium interactions and by pi-pi and van der Waals interactions. The rate of decomplexation of 2 is estimated to be 3.1 s(-1) from a 2D exchange NMR spectrum. The [60]fullerene encapsulation process can be controlled by modifying the amounts of acids used, changing the temperature of the system, altering the ratio of acid/base, and even through varying the solvent polarity. Moreover, the fluorescence spectra show band-narrowing spectral changes and a retardation of the relaxation characteristics of isolated and isotropic [60]fullerenes, which indicates that the environmental change around [60]fullerene is induced upon its encapsulation. PMID- 18431732 TI - The mechanism of the (bispidine)copper(II)-catalyzed aziridination of styrene: a combined experimental and theoretical study. AB - Experimental and DFT-based computational results on the aziridination mechanism and the catalytic activity of (bispidine)copper(I) and -copper(II) complexes are reported and discussed (bispidine=tetra- or pentadentate 3,7 diazabicyclo[3.1.1]nonane derivative with two or three aromatic N donors in addition to the two tertiary amines). There is a correlation between the redox potential of the copper(II/I) couple and the activity of the catalyst. The most active catalyst studied, which has the most positive redox potential among all (bispidine)copper(II) complexes, performs 180 turnovers in 30 min. A detailed hybrid density functional theory (DFT) study provides insight into the structure, spin state, and stability of reactive intermediates and transition states, the oxidation state of the copper center, and the denticity of the nitrene source. Among the possible pathways for the formation of the aziridine product, the stepwise formation of the two N-C bonds is shown to be preferred, which also follows from experimental results. Although the triplet state of the catalytically active copper nitrene is lowest in energy, the two possible spin states of the radical intermediate are practically degenerate, and there is a spin crossover at this stage because the triplet energy barrier to the singlet product is exceedingly high. PMID- 18431734 TI - Chiral bisphosphazides as dual basic enantioselective catalysts. AB - Chiral bisphosphazides complexed with lithium salts efficiently catalyze the direct enantioselective 1,4-addition of dialkyl malonates to acyclic enones. Spectroscopic studies on the stoichiometry of the bisphosphazide and lithium salt have indicated the formation of a 1:1 species as the active enantioselective catalyst. It is suggested that the catalyst generates a complex of the protonated phosphazide and the chiral nucleophile as the key intermediate. The phosphazide moiety appears to be a promising dual basic functionality for stereo- and chemoselective catalytic transformations. PMID- 18431733 TI - Cowpea mosaic virus capsid: a promising carrier for the development of carbohydrate based antitumor vaccines. AB - Immunotherapy targeting tumor cell surface carbohydrates is a promising approach for cancer treatment. However, the low immunogenecity of carbohydrates presents a formidable challenge. We describe here the enhancement of carbohydrate immunogenicity by an ordered display on the surface of the cowpea mosaic virus (CPMV) capsid. The Tn glycan, which is overexpressed on numerous cancer cell surfaces, was selected as the model antigen for our study. Previously it has been shown that it is difficult to induce a strong T cell-dependent immune response against the monomeric form of Tn presented in several ways on different carriers. In this study, we first synthesized Tn antigens derivatized with either a maleimide or a bromoacetamide moiety that was conjugated selectively to a cysteine mutant of CPMV. The glycoconjugate was then injected into mice and pre- and post-immune antibody levels in the mice sera were measured by enzyme-linked immunosorbant assays. High total antibody titers and, more importantly, high IgG titers specific for Tn were obtained in the post-immune day 35 serum, suggesting the induction of T cell-dependent antibody isotype switching by the glycoconjugate. The antibodies generated were able to recognize Tn antigens presented in their native conformations on the surfaces of both MCF-7 breast cancer cells and the multidrug resistant breast cancer cell line NCI-ADR RES. These results suggest that the CPMV capsid can greatly enhance the immunogenicity of weak antigens such as Tn and this can provide a promising tool for the development of carbohydrate based anti-cancer vaccines. PMID- 18431735 TI - Selective formation of 2-imidazolines and 2-substituted oxazoles by using a three component reaction. AB - Selective formation of 2H-2-imidazolines and 2-substituted oxazoles by using a multicomponent reaction of amines, either aldehydes or ketones, and alpha-acidic isocyano amides or esters is described. By selecting the appropriate solvent, Ag(I) or Cu(I) catalyst, or by employing a weak Bronsted acid, the product formation can be fully controlled and directed quantitatively to the desired heterocyclic scaffold. The described experimental procedures not only significantly increase the scope of compatible inputs for this complexity generating three-component reaction, but also allow for considerable chemical diversity: At least four diversity points in two distinct scaffolds can be exploited in this way. PMID- 18431736 TI - Novel bifunctional chiral urea and thiourea derivatives as organocatalysts: enantioselective nitro-Michael reaction of malonates and diketones. PMID- 18431737 TI - MLPA screening in the BRCA1 gene from 1,506 German hereditary breast cancer cases: novel deletions, frequent involvement of exon 17, and occurrence in single early-onset cases. AB - We present a comprehensive analysis of 1,506 German families for large genomic rearrangements (LGRs) in the BRCA1 gene and of 450 families in the BRCA2 gene by the multiplex ligation-dependent probe amplification (MLPA) technique. A total of 32 pathogenic rearrangements in the BRCA1 gene were found, accounting for 1.6% of all mutations, but for 9.6% of all BRCA1 mutations identified in a total of 1,996 families, including 490 with small pathogenic BRCA1/2 mutations. Considering only high risk groups for hereditary breast/ovarian cancer, the prevalence of rearrangements is 2.1%. Interestingly, deletions involving exon 17 of the BRCA1 gene seem to be most frequent in Germany. Apart from recurrent aberrations like del ex17, dupl ex13, and del ex22, accounting for more than 50% of all BRCA1 LGRs, we could fully characterize 11 novel deletions. Moreover, one novel deletion involving exons 1-7 and one deletion affecting the entire BRCA1 gene were identified. All rearrangements were detected in families with: 1) at least two breast cancer cases prior to the age of 51 years; 2) breast and ovarian cancer cases; 3) ovarian cancer only families with at least two ovarian cancer cases; or 4) a single breast cancer case prior to the age of 36 years, while no mutations were detected in breast cancer only families with no or only one breast cancer case prior to the age of 51 years. Analysis for gross rearrangements in 412 high-risk individuals, revealed no event in the BRCA2 gene and only two known CHEK2 mutations. However, in an additional 38 high-risk families with cooccurrence of female breast/ovarian and male breast cancer, one rearrangement in the BRCA2 gene was found. In summary, we advise restricting BRCA1 MLPA screening to those subgroups that revealed LGRs and recommend BRCA2 MLPA screening only for families presenting with cooccurrence of female and male breast cancer. PMID- 18431738 TI - Effects of weak mobile phone - electromagnetic fields (GSM, UMTS) on well-being and resting EEG. AB - Modern mobile phones emit electromagnetic fields (EMFs) ranging from 900 to 2000 MHz which are suggested to have an influence on well-being, attention and neurological parameters in mobile phone users. To date most studies have investigated Global System for Mobile Communications (GSM)-EMF and only very few studies were concerned with Universal Mobile Telecommunications System (UMTS) EMF. Consequently, we tested the effects of both types of EMF, 1950 MHz UMTS (SAR 0.1 and 1 W/kg) and pulsed 900 MHz GSM (1 W/kg), on well-being and vigilance controlled resting electroencephalogram (eyes closed) in 15 healthy, right-handed subjects. A double-blind, randomised, crossover application of the test procedure was used. Neither the UMTS- nor the GSM-EMF produced any significant changes in the measured parameters compared to sham exposure. The results do not give any evidence for a deleterious effect of the EMF on normal healthy mobile phone users. PMID- 18431739 TI - Cerebral hemorrhage: precipitating event for a tako-tsubo-like cardiomyopathy? AB - There have been some reports in medical literature of patients with profound, reversible left-ventricular (LV) dysfunction after sudden emotional stress. Originally, in Japan, the pattern was called tako-tsubo cardiomyopathy. ST segment elevation is seen with this syndrome, and it may look like acute ST elevation myocardial infarction. However, it has also been well established that a subarachnoid hemorrhage can manifest with cardiac abnormalities. The authors describe a case involving a cerebral hemorrhage and a potential tako-tsubo-like cardiomyopathy in the same patient. In conclusion, even though the patient's presentation appears to be a tako-tsubo syndrome, the radiographic evidence of a cerebral hemorrhage presents as a stronger etiology for the cardiac changes that occur in our patient. PMID- 18431740 TI - Two cases with similar pseudoaneurysms but different outcomes. AB - The pseudoaneurysm is a rare cardiac pathology, in which the left ventricular free wall ruptures and the pericardium surrounds the rupture in combination with thrombus and inflammation and thus prevents the development of a hemopericardium. Left ventricular pseudoaneurysm may remain silent unless it gives rise to cardiac tamponade, collapse, and finally sudden death. In this case report, we present two cases with left ventricular pseudoaneurysms in the same area. One of them had a stable outcome and the other had a fatal outcome. PMID- 18431741 TI - Transient left ventricular basal ballooning-"inverted" Tako-tsubo. PMID- 18431742 TI - Genistein mediated histone acetylation and demethylation activates tumor suppressor genes in prostate cancer cells. AB - Genistein is a phytoestrogen that has been reported to suppress the AKT signaling pathway in several malignancies. However, the molecular mechanism of genistein action is not known. We tested the hypothesis that genistein activates expression of several aberrantly silenced tumor suppressor genes (TSGs) that have unmethylated promoters such as PTEN, CYLD, p53 and FOXO3a. We report here that genistein activates TSGs through remodeling of the heterochromatic domains at promoters in prostate cancer cells by modulating histone H3-Lysine 9 (H3-K9) methylation and deacetylation. Genistein activation involved demethylation and acetylation of H3-K9 at the PTEN and the CYLD promoter, while acetylation of H3 K9 at the p53 and the FOXO3a promoter occurred through reduction of endogenous SIRT1 activity. There was a decrease of SIRT1 expression and accumulation of SIRT1 in the cytoplasm from the nucleus. Increased expression of these TSGs was also reciprocally related to attenuation of phosphorylated-AKT and NF-kappaB binding activity in prostate cancer cells. This is the first report describing a novel epigenetic pathway that activates TSGs by modulating either histone H3 Lysine 9 (H3-K9) methylation or deacetylation at gene promoters leading to inhibition of the AKT signaling pathway. These findings strengthen the understanding of how genistein may be chemoprotective in prostate cancer. PMID- 18431744 TI - Opportunities for renewable bioenergy using microorganisms. AB - Global warming can be slowed, and perhaps reversed, only when society replaces fossil fuels with renewable, carbon-neutral alternatives. The best option is bioenergy: the sun's energy is captured in biomass and converted to energy forms useful to modern society. To make a dent in global warming, bioenergy must be generated at a very high rate, since the world today uses approximately 10 TW of fossil-fuel energy. And, it must do so without inflicting serious damage on the environment or disrupting our food supply. While most bioenergy options fail on both counts, several microorganism-based options have the potential to produce large amounts of renewable energy without disruptions. In one approach, microbial communities convert the energy value of various biomass residuals to socially useful energy. Biomass residuals come from agricultural, animal, and a variety of industrial operations, as well as from human wastes. Microorganisms can convert almost all of the energy in these wastes to methane, hydrogen, and electricity. In a second approach, photosynthetic microorganisms convert sunlight into biodiesel. Certain algae (eukaryotes) or cyanobacteria (prokaryotes) have high lipid contents. Under proper conditions, these photosynthetic microorganisms can produce lipids for biodiesel with yields per unit area 100 times or more than possible with any plant system. In addition, the non-lipid biomass can be converted to methane, hydrogen, or electricity. Photosynthetic microorganisms do not require arable land, an advantage because our arable land must be used to produce food. Algae or cyanobacteria may be the best option to produce bioenergy at rates high enough to replace a substantial fraction of our society's use of fossil fuels. PMID- 18431745 TI - Structure analysis and performance of a microbial community from a contaminated aquifer involved in the complete reductive dechlorination of 1,1,2,2 tetrachloroethane to ethene. AB - An anaerobic microcosm set up with aquifer material from a 1,1,2,2 tetrachloroethane (TeCA) contaminated site and amended with butyrate showed a complete TeCA dechlorination to ethene. A structure analysis of the microbial community was performed by fluorescence in situ hybridization (FISH) with already available and on purpose designed probes from sequences retrieved through 16S rDNA clone library construction. FISH was chosen as identification tool to evaluate in situ whether the retrieved sequences belong to primary bacteria responsible for the biodegradative reactions. FISH probes identified up to 80% of total bacteria and revealed the absence or the marginal presence of known TeCA degraders and the abundance of two well-known H(2)-utilizing halorespiring bacteria, Sulfurospirillum (32.4 +/- 8.6% of total bacteria) and Dehalococcoides spp. (14.8 +/- 2.8), thereby providing a strong indication of their involvement in the dechlorination processes. These results were supported by the kinetic and thermodynamic analysis which provided indications that hydrogen was the actual electron donor for TeCA dechlorination. The specific probes, developed in this study, for known dechlorinators (i.e., Geobacter, Dehalobacter, and Sulfurospirillum species) represent a valuable tool for any future in situ bioremediation study as well as a quick and specific investigation tool for tracking their distribution in the field. PMID- 18431746 TI - High-flow priapism complicating ischemic priapism following iatrogenic laceration of the dorsal artery during a Winter procedure. AB - We report a case of a patient who developed high-flow priapism following a Winter procedure performed for the treatment of low-flow ischemic priapism. During the creation of bilateral cavernosal-glandular shunts, the tip of the left dorsal artery was accidentally lacerated. A fistula developed through the shunt between the torn artery and the cavernosal sinusoids of the tip of the left corpus cavernosum. Gray-scale and Doppler sonography and angiography allowed diagnosis and guided the therapeutic approach. PMID- 18431743 TI - Consortium analysis of 7 candidate SNPs for ovarian cancer. AB - The Ovarian Cancer Association Consortium selected 7 candidate single nucleotide polymorphisms (SNPs), for which there is evidence from previous studies of an association with variation in ovarian cancer or breast cancer risks. The SNPs selected for analysis were F31I (rs2273535) in AURKA, N372H (rs144848) in BRCA2, rs2854344 in intron 17 of RB1, rs2811712 5' flanking CDKN2A, rs523349 in the 3' UTR of SRD5A2, D302H (rs1045485) in CASP8 and L10P (rs1982073) in TGFB1. Fourteen studies genotyped 4,624 invasive epithelial ovarian cancer cases and 8,113 controls of white non-Hispanic origin. A marginally significant association was found for RB1 when all studies were included [ordinal odds ratio (OR) 0.88 (95% confidence interval (CI) 0.79-1.00) p = 0.041 and dominant OR 0.87 (95% CI 0.76 0.98) p = 0.025]; when the studies that originally suggested an association were excluded, the result was suggestive although no longer statistically significant (ordinal OR 0.92, 95% CI 0.79-1.06). This SNP has also been shown to have an association with decreased risk in breast cancer. There was a suggestion of an association for AURKA, when one study that caused significant study heterogeneity was excluded [ordinal OR 1.10 (95% CI 1.01-1.20) p = 0.027; dominant OR 1.12 (95% CI 1.01-1.24) p = 0.03]. The other 5 SNPs in BRCA2, CDKN2A, SRD5A2, CASP8 and TGFB1 showed no association with ovarian cancer risk; given the large sample size, these results can also be considered to be informative. These null results for SNPs identified from relatively large initial studies shows the importance of replicating associations by a consortium approach. PMID- 18431747 TI - Intramuscular and subcutaneous sparganosis: Sonographic findings. AB - A case of subcutaneous and intramuscular sparganosis was confirmed on surgical excision of a worm in a 60-year-old woman suffering from painful masses in the right thigh. Sonography and MRI revealed an ill-defined intramuscular lesion and multiple cystic lesions in the subcutaneous tissue. At the time of the excision, a sparganum larva was found in the adductor longus muscle. If an intramuscular mass with a serpiginous cystic tract is seen on imaging studies in an endemic area, musculoskeletal sparganosis should be included in the differential diagnosis of a soft tissue tumor. PMID- 18431748 TI - Hypospadias mimicking female genitalia on early second trimester sonographic examination. AB - Female fetal sex was assigned based on downward direction of the genital tubercle at 14 weeks' gestation. Subsequently, amniocentesis performed due to the finding of an echogenic focus in the left ventricle revealed a male karyotype. Sonographic examination at 22 weeks' gestation revealed a markedly curved penile shaft. Power Doppler at that time demonstrated a urine jet emanating from the penile base, verifying the diagnosis of penoscrotal hypospadias. We therefore suggest that female fetal sex assignment should be based on the demonstration of the 2 or 4 parallel lines representing the labial folds and not only upon the demonstration of the downward direction of the genital tubercle. PMID- 18431749 TI - Sonography of dermatofibrosarcoma protuberans of the groin. AB - Dermatofibrosarcoma protuberans (DFSP) is an uncommon tumor that arises from the dermis of adults. Its malignant potential is considered to be intermediate. Because the diagnosis of DFSP can frequently be suspected on the basis of physical examination reports on its sonographic appearance are rare. We report a case that highlights the value of sonography in assessing the subcutaneous extent of DFSP. PMID- 18431750 TI - Solution NMR structure of the SOS response protein YnzC from Bacillus subtilis. PMID- 18431751 TI - Crystal structure of Tflp: a ferredoxin-like metallo-beta-lactamase superfamily protein from Thermoanaerobacter tengcongensis. PMID- 18431752 TI - Development of dual-compartment perfusion bioreactor for serial coculture of hepatocytes and stellate cells in poly(lactic-co-glycolic acid)-collagen scaffolds. AB - An important challenge in liver tissue engineering is to overcome the rapid loss of hepatocyte functions. In vivo, hepatocytes are compact polyhedral cells with round nuclei; however, they readily loss many of their differentiated functions in vitro. To overcome this challenge, we have established a new perfusion bioreactor that consists of two compartments which enabled the serial coculture of hepatocytes and hepatic stellate cells-T6 without direct contact between each other. Three dimensional scaffolds were utilized in the bioreactor as physical anchors for cells. The scaffolds consist of collagen grafted poly(lactic-co glycolic acid) microfibers and cross-linked collagen sponges between microfibers for additional cellular support and adhesion. The advantages of this new bioreactor are enabling cell culture in three dimensional organization and controlling the culture parameters of the supporting cells independently from the hepatocytes. The results showed that the hepatocytes exhibited much higher levels of the differentiated functions such as albumin secretion, urea synthesis, and cytochrome P450 enzymatic activity when compared with the monoculture system where hepatocytes alone were cultured. This perfusion bioreactor system has potential applications in the development of bioartificial liver devices or cell based tissue constructs transplantation therapies. PMID- 18431753 TI - Osteogenic differentiation of mesenchymal stem cells in defined protein beads. AB - There is a need to develop improved methods for directing and maintaining the differentiation of human mesenchymal stem cells (hMSC) for regenerative medicine. Here, we present a method for embedding cells in defined protein microenvironments for the directed osteogenic differentiation of hMSC. Composite matrices of collagen I and agarose were produced by emulsification and simultaneous polymerization in the presence of hMSC to produce 30-150 mum diameter hydrogel "beads." The proliferation, morphology, osteogenic gene expression, and calcium deposition of hMSC in bead environments were compared to other two- and three-dimensional culture environments over 14-21 days in culture. Cells embedded within 40% collagen beads exhibited equivalent proliferation rates to those in gel disks, but showed upregulation of bone sialoprotein and increased calcium deposition over 2D controls. Osteocalcin gene expression was not changed in 3D beads and disks, while collagen type I gene expression was downregulated relative to cells in 2D culture. The hydrogel bead format allows controlled cell differentiation and is a cell delivery vehicle that may also enhance vascular invasion and host incorporation. Our results indicate that the application of such beads can be used to promote the osteogenic phenotype in hMSC, which is an important step toward using them in bone repair applications. PMID- 18431754 TI - Material properties and bone marrow stromal cells response to in situ crosslinkable RGD-functionlized lactide-co-glycolide scaffolds. AB - In situ crosslinkable biomaterials with degradation profiles that can be tailored to a particular application are indispensable for treating irregularly shaped defects and for fabrication of shape-selective scaffolds. The objective of this work was to synthesize ultra low molecular weight functionalized PLA and PLGA macromers that can be grafted with bioactive peptides and crosslinked in situ to fabricate biodegradable functional scaffolds. In situ crosslinkable lactide-co glycolide macromer (cMLGA; "c" for crosslinkable, "M" for macromer, and "LGA" for lactide-co-glycolide) was synthesized by anionic polymerization of lactide and glycolide monomers followed by condensation polymerization with fumaryl chloride. The cMLA (100% L-lactide) and cMLGA macromers formed porous crosslinked scaffolds with NVP as the crosslinker. The mass loss of the crosslinked cMLA and cMLGA was linear with incubation time in vitro (zero-order degradation) and the degradation rate depended on the ratio of lactide to glycolide. cMLGA scaffold with 1:1 lactide to glycolide ratio completely degraded after 4 weeks while the cMLA lost less than 40% of its initial mass after 35 weeks. When cMLA scaffold was functionalized with acrylated integrin-binding Ac-GRGD amino acid sequence, bone marrow stromal (BMS) cells attached and spread on the cMLA scaffold and exhibited focal-point cell adhesion. The mRNA expression levels of collagen-1alpha, osteonectin, and osteopontin for BMS cells seeded in the scaffolds with 1 and 5% Ac-GRGD were upregulated compared with those without Ac-GRGD. cMLGA is attractive as in situ crosslinkable macromer for fabrication of functional scaffolds with degradation characteristics that can be tailored to a particular application. PMID- 18431755 TI - A new generation of high flex life polyurethane urea for polymer heart valve- studies on in vivo biocompatibility and biodurability. AB - Development of new generation high flex life polyurethane urea (HFL18-PU) with appropriate elastic modulus, biocompatibility, blood compatibility, resistant to calcification, and biodurability for the long-term use as cardiac device is still a challenge. This study reports the development of a fully aliphatic, ether-free physically cross-linked and low elastic modulus (6.841 +/- 0.27 MPa) polyurethane urea having in vivo biostability, in vivo biocompatibility and high flex-life (721 +/- 30 million cycles) that can satisfy the requirements for the fabrication of tri-leaflet heart valve. PMID- 18431756 TI - Bond strength of adhesively luted ceramic discs to different core materials. AB - The purpose of this in vitro study was to compare the shear bond strengths of resin, glass-ionomer, and ceramic-based core materials to all ceramic discs. Five core materials (Core max, Sankin; Clearfil AP-X, Kuraray; Empress Cosmo, Ivoclar Vivadent; Photocore, Kuraray; Dyract Extra, Dentsply) were prepared as discs 10 mm in diameter and 2 mm in height according to the manufacturer's instructions. Ten disc specimens per group were prepared, and dentin served as the control. All resin specimens were embedded in autopolymerizing acrylic resin, with one surface facing up. All ceramic discs (IPS Empress I, Ivoclar-Vivadent) 3 mm in diameter and 2 mm in height were prepared and bonded to core specimens with a dual-curing luting resin cement (Variolink II, Vivadent). Specimens were stored in distilled water at 37 degrees C. Shear bond strength of each sample was measured after 24 h using a universal testing machine at a crosshead speed of 0.5 mm/min. The data were analyzed with one-way analysis of variance and Tukey HSD tests (alpha = 0.05). Shear bond strength varied significantly depending on the core material used (p < 0.05). Clearfil AP-X and Photocore showed the highest shear bond strength value while Empress Cosmo provided the lowest (p < 0.05). There were no statistically significant differences among Clearfil AP-X, Photocore, and Core Max (p > 0.05). And also there were no statistically significant differences between Dyract Extra and the control group (p > 0.05). In vitro shear bond strengths of ceramic discs bonded to resin-based core materials showed higher bond strength values than ceramic-based core material. PMID- 18431757 TI - Nickel release behavior and surface characteristics of porous NiTi shape memory alloy modified by different chemical processes. AB - As a non-line-of-sight surface modification technique, chemical treatment is an effective method to treat porous NiTi with complex surface morphologies and large exposed areas due to its liquidity and low temperature. In the work described here, three different chemical processes are used to treat porous NiTi alloys. Our results show that H(2)O(2) treatment, NaOH treatment, and H(2)O(2) pre treatment plus subsequent NaOH treatment can mitigate leaching of nickel from the alloy. The porous NiTi samples modified by the two latter processes favor deposition of a layer composed of Ca and P due to the formation of bioactive Na(2)TiO(3) on the surface. Among the three processes, H(2)O(2) pre-treatment plus subsequent NaOH modification is the most effective in suppressing nickel release. Small area X-ray photoelectron spectroscopy reveals that the surfaces treated by different chemical processes have different structures and compositions. The sample modified by the H(2)O(2) treatment is composed of rough TiO(2) on the outer surface and an oxide transition layer underneath whereas the sample treated by NaOH comprises a surface layer of titanium oxide and Na(2)TiO(3) together with a transition layer. The sample processed by the H(2)O(2) and NaOH treatment has a pure Na(2)TiO(3) layer on the surface and a transition layer underneath. These results help to elucidate the different nickel release behavior and bioactivity of porous NiTi alloys processed by different methods. PMID- 18431758 TI - Fabrication and characterization of novel nano- and micro-HA/PCL composite scaffolds using a modified rapid prototyping process. AB - Novel three-dimensional scaffolds consisting of nano- and microsized hydroxyapatite (HA)/poly(epsilon-caprolactone) (PCL) composite were fabricated using a modified rapid-prototyping (RP) technique for bone tissue engineering applications. The size of the nano-HA ranged from 20 to 90 nm, whereas that of the micro-HA ranged from 20 to 80 microm. The scaffold macropores were well interconnected, with a porosity of 72-73% and a pore size of 500 microm. The compressive modulus of the nano-HA/PCL and micro-HA/PCL scaffolds was 3.187 +/- 0.06 and 1.345 +/- 0.05 MPa, respectively. The higher modulus of the nano-HA/PCL composite (n-HPC) was to be likely caused by a dispersion strengthening effect. The attachment and proliferation of MG-63 cells on n-HPC were better than that on the micro-HA/PCL composite (m-HPC) scaffold. The n-HPC was more hydrophilic than the m-HPC because of the greater surface area of HA exposed to the scaffold surface. This may give rise to better cell attachment and proliferation. Bioactive n-HA/PCL composite scaffold prepared using a modified RP technique has a potential application in bone tissue engineering. PMID- 18431760 TI - Bacterial adherence to SiO2-based multifunctional bioceramics. AB - The bacterial adherence onto different multifunctional silica-based bioceramics has been evaluated. Staphylococcus aureus and Staphylococcus epidermidis were chosen, as they cause the majority of the implant-related infections in this field. Two SiO2 mesoporous materials (MCM-41, SBA-15), an ordered SiO2-CaO-P2O5 mesoporous glass (OMG), and a biphasic magnetic bioceramic (BMB), were incubated with S. aureus and S. epidermidis for 90 min, and subsequently sonicated to quantify the number of adhered bacteria on each material. It was found that S. aureus and S. epidermidis (10(8) CFU/mL) adhered significantly less to BMB samples when compared to MCM-41, SBA-15, or OMG. However, when the material pores accessible for bacteria in each material were taken into account, the lowest bacterial adherence was found in MCM-41, and the highest in SBA-15. The results show that bacterial adherence is higher on mesoporous bioceramics, although this higher microbial attachment is mainly due to the intergranular porosity and grain size morphology rather than to the mesoporous structure. PMID- 18431759 TI - Quantitative in vivo cytokine analysis at synthetic biomaterial implant sites. AB - To further elucidate the foreign body reaction, investigation of cytokines at biomaterial implant sites was carried out using a multiplex immunoassay and ELISA. Macrophage activation cytokines (IL-1beta, IL-6, and TNFalpha), cytokines important for macrophage fusion (IL-4 and IL-13), antiinflammatory cytokines (IL 10 and TGFbeta), chemokines (GRO/KC, MCP-1), and the T-cell activation cytokine IL-2 were quantified at biomaterial implant sites. Empty cages (controls) or cages containing synthetic biomedical polymer (Elasthane 80A (PEU), silicone rubber (SR), or polyethylene terephthalate (PET)) were implanted subcutaneously in Sprague-Dawley rats for 4, 7, or 14 days, and cytokines in exudate supernatants and macrophage surface adhesion and fusion were quantified. The presence of a polymer implant did not affect the levels of IL-1beta, TGFbeta, and MCP-1 in comparison to the control group. IL-2 was not virtually detected in any of the samples. Although the levels of IL-4, IL-13, IL-10, and GRO/KC were affected by polymer implantation, but not dependent on a specific polymer, IL-6 and TNFalpha were significantly greater in those animals implanted with PEU and SR, materials that do not promote fusion. The results indicate that differential material-dependent cytokine profiles are produced by surface adherent macrophages and foreign body giant cells in vivo. PMID- 18431761 TI - Biological response to micron- and nanometer-sized particles known as potential wear products from artificial hip joints: Part I: Selection and characterization of model particles. AB - The aim of this work was to select and characterize model particles, which correspond to real wear products from artificial hip joints, and to investigate the dispersing behavior of these powders. Commercially available nano and microparticles of corundum, graphite, and chromium oxide were selected or alternatively self-produced by milling. These powders were characterized regarding density, specific surface area, crystalline phases, particle size distributions and shape. Volume-based particle size distributions Q(3)(d) were measured after dispersing in water, water with dispersant, Ringers solution, and cell culture solution (Dulbecco's Modified Eagle's Medium (DMEM)) by laser diffraction and ultrasonic spectroscopy. Nanopowders formed agglomerates in the micrometer range in cell culture solutions. The micropowders showed only a marginal agglomeration. The median diameters of the dispersed nanopowders were even bigger than those of micropowders. Calculations of the number-based size distribution Q(0)(d) showed that in spite of the agglomeration the predominant number of the nano and microparticles is in the sub micrometer range, with only one exception, the micrographite powder. PMID- 18431762 TI - Apparent conversion of adhesive resin in the hybrid layer, Part II: In situ studies of the resin-dentin bond. AB - Penetration and conversion of adhesives into the hybrid layer (HL) is important to the quality and longevity of the adhesive resin (AR)-dentin bond. In this study, a methodology is developed to examine the degree of conversion and relative HEMA concentration with respect to Bis-GMA using Raman spectroscopy. This methodology will be used in the future reports related to this topic. Conversion in the AR of water-stored resin-dentin samples (84% +/- 3%) agreed well with that measured in commercial adhesive (Comm Adh) resin samples after 24 h water storage (80% +/- 2% from Part 1) and was significantly higher than Comm Adh without water storage (58% +/- 3% from Part 1) (p = 0.0005). Adhesive conversion was not significantly different (p = 0.5036) through the middle of the HL, with a mean of 83% +/- 6%. HEMA mole fraction, relative to Bis-GMA, was significantly higher (p = 0.0028) in the top half of the HL (0.67 +/- 0.03), when compared to HEMA in the AR (0.60 +/- 0.01). HEMA and EDMAB were identified through GC/MS as leachable components in the aqueous 24-h storage media. The effect of this elution explains the change in conversion measurements observed between dry and water-stored conditions, which is more appropriately described as the "apparent" conversion. PMID- 18431763 TI - The effect of dehydrothermal treatment on the mechanical and structural properties of collagen-GAG scaffolds. AB - The mechanical properties of tissue engineering scaffolds are critical for preserving the structural integrity and functionality during both in vivo implantation and long-term performance. In addition, the mechanical and structural properties of the scaffold can direct cellular activity within a tissue-engineered construct. In this context, the aim of this study was to investigate the effects of dehydrothermal (DHT) treatment on the mechanical and structural properties of collagen-glycosaminoglycan (CG) scaffolds. Temperature (105-180 degrees C) and exposure period (24-120 h) of DHT treatment were varied to determine their effect on the mechanical properties, crosslinking density, and denaturation of CG scaffolds. As expected, increasing the temperature and duration of DHT treatment resulted in an increase in the mechanical properties. Compressive properties increased up to twofold, while tensile properties increased up to 3.8-fold. Crosslink density was found to increase with DHT temperature but not exposure period. Denaturation also increased with DHT temperature and exposure period, ranging from 25% to 60% denaturation. Crosslink density was found to be correlated with compressive modulus, whilst denaturation was found to correlate with tensile modulus. Taken together, these results indicate that DHT treatment is a viable technique for altering the mechanical properties of CG scaffolds. The enhanced mechanical properties of DHT-treated CG scaffolds improve their suitability for use both in vitro and in vivo. In addition, this work facilitates the investigation of the effects of mechanical properties and denaturation on cell activity in a 3D environment. PMID- 18431764 TI - Efficacy of a p38 mitogen activated protein kinase inhibitor in mitigating an established inflammatory reaction to polyethylene particles in vivo. AB - The inhibitor of p38 mitogen-activated protein kinase (MAPK) is of interest in the nonoperative treatment of periprosthetic osteolysis due to wear particles. Previous studies demonstrated that an oral p38 MAPK inhibitor did not suppress bone formation when given during the initial phase of tissue differentiation. However, the oral p38 MAPK inhibitor also did not curtail the foreign body and chronic inflammatory response to particles when given simultaneously. The purpose of the current study was to examine the efficacy of a p38 MAPK inhibitor, SCIO 323, on mitigating an established inflammatory reaction that parallels the clinical situation more closely. The Bone Harvest Chamber was implanted in rabbits and submicron polyethylene particles were placed in the chamber for 6 weeks. The contents of the chambers were harvested every 6 weeks. Oral treatment with the SCIO-323 included delivery for 3 weeks and stopping for 3 weeks, delivery for 3 weeks after an initial 3-week delay, and delivery for 6 weeks continuously. Administration of the SCIO-323 continuously for 6 weeks with/without the presence of particles, or for the initial 3 of 6 weeks had minor effects on bone ingrowth. After establishing a particle-induced chronic inflammatory reaction for 3 weeks, administration of SCIO-323 for a subsequent 3 weeks suppressed net bone formation. The activity of osteoclast-like cells remained low among all treatments when compared with the first control. Using the present model, the oral p38 MAPK inhibitor was ineffective in improving bone ingrowth in the presence of polyethylene particles. PMID- 18431765 TI - Doubly stochastic Poisson distribution of platelet adhesion on material surfaces and its implication on fluorescence image analysis. AB - An image based assay has been developed to quantify platelet adhesion on material surfaces. Briefly, citrated platelet rich plasma (PRP) is incubated with materials for 2 h to allow platelet adhesion on the surface, followed by fluorescence labeling of platelets with Celltracker Green. Multiple images are acquired by an automatic fluorescence microscope, IN Cell Analyzer 1000. Platelets are identified and counted by an automatic image analysis algorithm. We have observed that the variance of the counts is considerably greater than expected from simple distribution laws. Statistical analysis of that difference shows that these measurements will often follow a doubly stochastic Poisson process in which the variance is inherently very large. To overcome this, multiple images (n > or = 8 images/well, about 3% of total area) are necessary to achieve accurate counting. This method has been compared to the commonly used enzyme based platelet adhesion assay, lactate dehydrogenase (LDH) assay. It is concluded that the present method is only effective in quantifying adherent platelets when a large number of samples are used. However, this method does provide additional information on platelet morphology and spatial distribution, which is lacking in the LDH assay. PMID- 18431766 TI - Evaluation of antitubercular drug-loaded surfactants as inhalable drug-delivery systems for pulmonary tuberculosis. AB - Pulmonary tuberculosis is associated with a year-long chemotherapy, poor alveolar drug levels, drug-related systemic toxicity, and patient noncompliance. In this study, exogenous pulmonary surfactant is proposed as a drug carrier for antitubercular drugs. Dipalmitoylphosphatidylcholine (DPPC), the major lung surfactant lipid, has been combined with antitubercular drugs isoniazid (INH), rifampicin (RFM), and ethambutol (ETH) in 1:1 ratio by weight, in which drugs had a ratio of 1:2:3 by weight. At 37 degrees C, the formulation had better surfactant function with quicker reduction of surface tension on adsorption (32.71 +/- 0.65 mN/m) than DPPC liposomes (44.67 +/- 0.57 mN/m) and maintained 100% airway patency in a capillary surfactometer. Drug-loaded surfactant liposomes were about 2 microm and had entrapment efficiency of 30.04% +/- 2.05%, 18.85% +/- 2.92%, and 61.47% +/- 3.32% for INH, RFM, and ETH, respectively. Sustained release of the drugs from surfactants was observed over 24 h. In vitro alveolar deposition efficiency using the twin impinger showed 12.06% +/- 1.87% of INH, 43.30% +/- 0.87% of RFM, and 22.07% +/- 2.02% of ETH deposited in the alveolar chamber upon nebulization for a minute using a jet nebulizer. The formulation was biocompatible and stable with physicochemical properties being retained even after storage for a month at 4 degrees C. Antitubercular drug loaded surfactants developed could serve dual purposes of alveolar stabilization due to surfactant action and better reach of these drugs to the alveoli due to antiatelectatic effect of the surfactant. PMID- 18431767 TI - Nasal chondrocytes and fibrin sealant for cartilage tissue engineering. AB - Hybrid constructs associating a biodegradable matrix and autologous chondrocytes hold promise for the treatment of articular cartilage defects. In this context, our objective was to investigate the potential use of nasal chondrocytes associated with a fibrin sealant for the treatment of articular cartilage defects. The phenotype of primary nasal chondrocytes (NC) from human (HNC) and rabbit (RNC) origin were characterized by RT-PCR. The ability of constructs associating fibrin sealant and NC to form a cartilaginous tissue in vivo was investigated, firstly in a subcutaneous site in nude mice and secondly in an articular cartilage defect in rabbit. HNC express type II collagen and aggrecan, the two major hallmarks of a chondrocytic phenotype. Furthermore, when injected subcutaneously into nude mice within a fibrin sealant, these chondrocytes were able to form a cartilage-like tissue. Our data indicate that RNC also express type II collagen and aggrecan and maintained their phenotype in three-dimensional culture within a fibrin sealant. Moreover, treatment of rabbit articular cartilage defects with autologous RNC embedded in a fibrin sealant led to the formation of a hyalin-like repair tissue. The use of fibrin sealant containing hybrid autologous NC therefore appears as a promising approach for cell-based therapy of articular cartilage. PMID- 18431768 TI - Response of human osteoblasts exposed to wear particles generated at the interface of total hip stems and bone cement. AB - Aseptic loosening of total hip replacement is mainly caused by wear particles. Abrasive wear occurs at articulating surfaces or as a consequence of micro motions at the interface between femoral stem and bone cement. Direct impact of wear particles on osteolysis, the remodeling of the bone stock and a directly affected function of osteoblasts was described. The present study examined the response of human osteoblasts exposed to different wear particles, which were generated in a test device providing oscillating micro-motions at the interface between femoral stem and standard bone cement. Characterization of released particles was performed by quantifying the size distribution and the metal content of the wear debris. Human osteoblasts were incubated with particles obtained from hip stems with different material compositions (Ti-6Al-7Nb and Co 28Cr-6Mo) and rough and smooth surface finishings combined with standard bone cement (Palacos(R) R) containing zirconium oxide particles. Commercially pure titanium particles (cp-Ti) and particulate zirconium oxide (ZrO(2)) were used for comparative analyses. The results revealed significant (p < 0.05) reduction of the cell viability after exposure to higher concentration of metallic particles, particularly from Co-based alloys. In contrast, ZrO(2) alone showed significantly less adverse effects on the cells. When increasing metallic particle concentrations massive inhibition was also observed in the release of cytokines including interleukine-6 (IL-6) and interleukine-8 (IL-8), but the expression of Procollagen I and the cell viability showed the highest reduction after exposure to Co-based alloy particles from rough stems. PMID- 18431769 TI - Efficiency of controlled topical delivery of silver sulfadiazine in infected burn wounds. AB - The present study is designed to assess the potential benefits of controlled delivery of silver sulfadiazine from collagen scaffold (SSDM-CS) in infected deep partial thickness burn wounds in which epidermis is lost completely and the entire papillary dermis and most of the recticular layer of the dermis is lost. Infection induced by inoculating 10(7) colony forming units (cfu) of Pseudomonas aeruginosa caused significant increase in wound size (20%) till day 15, which decreased significantly from day 9 by SSDM-CS treatment, showing complete healing by day 27 (control > or = 37 days). Early subsidence of infection (<10(2) cfu, day 9) by SSDM-CS resulted in faster epidermal resurfacing and fibroplasia, whereas heavy microbial load (>10(7) cfu, day 9) in controls caused severe inflammatory cellular infiltration. Persistent infection triggered early expression of proinflammatory cytokines intereukin-6, intereukin 1-beta, and tumor necrosis factor-alpha, lasting until day 9, whereas cytokine level decreased in SSDM-CS-treated group by day 6. Infection exacerbated expression of active matrix metalloproteinases (MMPs)-2 and -9 in controls (day 15), while SSDM CS positively modulated MMP-2 and -9 with faster decline in their levels (day 12). Inherent nature of the dressing to maintain drug level at equilibrium therapeutic concentration (51.2 microg/mL) for prolonged time (72 h), below systemic toxic limits (20 microg/dL, serum level), accelerated the magnitude and sequence of reparative events. PMID- 18431770 TI - Energy consumption as a predictor test of the durability of a biological tissue employed in cardiac bioprosthesis. AB - The mechanical behavior of the young bull pericardium in a fatigue test has been studied. This material is a similar tissue to those used in valve leaflet construction for a cardiac bioprosthesis. The consumed energy on each test was evaluated and afterwards used as a predictor of the biomaterial strength. Two hundred and nine samples were tested to cyclical fatigue. The cut-off point to determine the sample quality was whether or not they resisted at least 4500 cycles. Only 22 samples withstood over that point (10.52%). The samples were classified according to their fatigue behavior in excellent, undefined and unsuitable. By using as a reference the consumed energy in the first 25 cycles, we could distinguish correctly (between 93.2 and 96.1%) the unsuitable material and most of the excellent (between 78.1 and 95.2%). From the rejected material 77% was really detachable and from the accepted, only 50% was excellent, with an equal methodology. The receiver operating characteristics curve was employed to establish decision levels when selecting samples, being 0.85 the best area (theoretical maximum value of 1). It is concluded that the energy wasted is a good predictor of the strength of the tissue. More than 90% of the unsuitable material and 50% of the excellent material (5% of all the material) is detected with this method. PMID- 18431771 TI - Interactions of coronary artery smooth muscle cells with 3D porous polyurethane scaffolds. AB - One strategy in vascular tissue engineering is the design of hybrid vascular substitutes where vascular cells infiltrate biostable porous scaffolds that provides favorable environment for guided cell repopulation and acts as a mechanically supporting layer after the tissue regeneration process. The aim of the present work was to study the interaction of human coronary artery smooth muscle cells (HCASMC) with 3D porous polyurethane scaffolds. We therefore fabricated porous and highly interconnected 3D polyurethane scaffolds that can promote HCASMC attachment, proliferation, and migration. SEM and microCT studies of the fabricated scaffolds showed that the current scaffolds had highly open and interconnected pore structures, with an average porosity of 84%. HCASMC interaction on polyurethane films revealed that cells adhere and express specific marker proteins (vinculin and h-caldesmon). This expression was further enhanced by coating the polyurethane with Matrigel. On uncoated 3D scaffolds, dense spherical aggregates of cells were often encountered with little adhesion of individual cells alongside the struts of the scaffold, independent of the porogens used. In contrast, when cultured on Matrigel-coated scaffolds, cell numbers quickly increased after 14 days and spread along the entire scaffold. At the upper scaffold surface, elongated cells were seen adhering to one another and also to the scaffold surface. These cells were elongated, aligned in parallel and contained abundant F-actin bundles suggesting a differentiated contractile phenotype. Deep into the scaffold, cells were encountered that formed actin-rich lamellipodial extensions spreading along the strut and lacked stress fibers, suggesting active cell migration along the substrate. PMID- 18431772 TI - Encapsulated Staphylococcus aureus strains vary in adhesiveness assessed by atomic force microscopy. AB - Staphylococcus aureus capsular polysaccharides are believed to play a role in adhesion to surfaces and may contribute to their antimicrobial resistance, thereby increasing the rates and severity of associated infections. The purpose of this study was to compare the adhesiveness of distinct S. aureus capsular polysaccharides to determine whether adhesiveness was a general or specific feature across different S. aureus strains. Atomic force microscopy was used to confirm the presence or absence of capsular polysaccharides and to measure adhesive forces on a noncapsulated, serotype 8, and serotype 2 strain of S. aureus. Serotype 8 displayed a larger range of adhesive forces (1-19 nN) than the noncapsulated (0-4 nN) and serotype 2 (0-4 nN) strain. The majority of adhesive forces for serotype 8 were in the 10-15 nN range. Removal of capsular polysaccharides gave a marked decrease in adhesive forces measured for serotype 8 and, to a lesser extent, a decrease for serotype 2. Noncapsulated, serotype 8, and serotype 2 S. aureus had water contact angles of 23.8 (+/-8.9), 34.4 (+/ 2.5), and 56.7 (+/-11.2) degrees (mean +/- standard deviation), respectively. For the first time, capsular polysaccharides from serotype 8 (clinically common) and serotype 2 (clinically rare) were demonstrated to have different physical properties, which may account for variations in studies in which clinical isolates are utilized, and the conflict in proposed roles for capsular polysaccharides on S. aureus is explained. PMID- 18431773 TI - Resorbable polymeric scaffolds for bone tissue engineering: the influence of their microstructure on the growth of human osteoblast-like MG 63 cells. AB - Degradable three-dimensional porous scaffolds applicable as cell carriers for bone tissue engineering were developed by an innovative solvent casting/particulate leaching technique from poly(L-lactide-co-glycolide) (PLG). Three types of PLG scaffolds were prepared, and these had the same high porosity (83%) but increasing diameter of the pores (180-200 microm, 250-320 microm, and 400-600 microm) and increasing pore interconnectivity. The colonization of the scaffolds with human osteoblast-like MG 63 cells was then studied in vitro in a conventional static cell culture system. The number of cells growing on the scaffolds on days 1 and 7 after seeding was highest in the material with the largest pore diameter, but on day 15, the differences among the scaffolds disappeared. Confocal microscopy revealed that on day 1 after seeding, the cells penetrated to a depth of 490 +/- 100 microm, 720 +/- 170 microm, and 720 +/- 120 microm into the scaffolds of small, medium, and large pore size, respectively. Incorporation of bromodeoxyuridine into newly synthesized DNA and the concentration of vinculin, beta-actin, osteopontin, and osteocalcin in cells on the scaffolds of all pore sizes were similar to the values obtained on standard tissue culture polystyrene, which indicated good biocompatibility of the scaffolds. These results suggest that all scaffolds could serve as good carriers for bone cells, although the quickest colonization with cells was found in the scaffolds with the largest pore diameter from 400 to 600 microm. PMID- 18431774 TI - Flow cytometry in plant breeding. AB - Since the first report on the flow cytometric study of plant material 35 years ago, analyzing the nuclear DNA content of field bean, an ever increasing number of applications of FCM has been developed and applied in plant science and industry, but a similar length of time elapsed before the appearance of the first complete volume devoted to FCM of plant cells. Most published information on the uses of FCM addresses various aspects of animal (including human) cell biology, thus failing to provide a pertinent substitute. FCM represents an ideal means for the analysis of both cells and subcellular particles, with a potentially large number of parameters analyzed both rapidly, simultaneously, and quantitatively, thereby furnishing statistically exploitable data and allowing for an accurate and facilitated detection of subpopulations. It is, indeed, the summation of these facts that has established FCM as an important, and sometimes essential, tool for the understanding of fundamental mechanisms and processes underlying plant growth, development, and function. In this review, special attention is paid to FCM as applied to plant cells in the context of plant breeding, and some new and less well-known uses of it for plants will be discussed. PMID- 18431775 TI - Automation and validation of a rapid method to assess neutrophil and monocyte activation by routine fluorescence flow cytometry in vitro. AB - The aim of the present study was to design an automated-gating hematology fluorescence flow cytometry methodology permitting the assessment of neutrophil and monocyte activation in EDTA-anticoagulated whole blood based on cell granularity, lipid membrane components, cell shape and volume, and total cell nucleic acid (NA) compounds. For particularly monitoring the proper functioning of patients' innate immune system as the first line defense against microbial invaders, the suitable test system should be rapid, simple, reliable by yielding reproducible results. It must be validated against established methods, and it must prove to work in selected clinical settings, e.g. in intensive care unit (ICU) environments. The adaptation of a routine hematology cell analyser utilizing fluorescence flow cytometry resulted in a potentially useful system for all requirements. It proved to detect in real-time and in a reliable and reproducible way the main cellular response reactions of neutrophils and monocytes during externally stimulated immune defense. Validation was successful when comparing it to established methods. The quantified activation effects were dose dependent from the applied activating agents. Cellular response kinetics could be measured and described and showed to be in line with the prevailing cell response models. Upon applying the test method to a healthy population of volunteers and a first cohort of ICU patients with and without evident immune depression, the test revealed excellent cellular responses to external activating cytotoxic stimuli (lipopolysaccharide; LPS) for the control group, slightly weaker response from ICU patients without immune depression and no response from patients with evident immune depression.We conclude that routine hematology fluorescence flow cytometry can accurately and reproducibly measure different activation steps of monocytes and polymorphonuclear neutrophilic granulocytes to defined external stimuli. This may potentially be applied as a STAT (Latin statim = immediately) and routine screening and surveillance method for inflammatory diseases. PMID- 18431776 TI - In vitro characterization of hepatocyte growth factor release from PHBV/PLGA microsphere scaffold. AB - Polymer scaffolds which can support cells to grow as well as deliver growth factors to the cells simultaneously have great potential for the successful regeneration of failed tissues. As popularly used vehicles to deliver anti-cancer drugs and growth factors, microspheres also show many advantages as substrates to guide the growth of cells. Therefore, we aimed to examine the feasibility of using microspheres as ideal scaffolds for liver tissue engineering. To determine the capabilities of previously used microsphere scaffold to deliver growth factors simultaneously, this work investigated a long-term (about three months) release of bovine serum albumin (BSA) from microsphere scaffolds fabricated by using two different polymers, poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV, 8% PHV), poly(lactide-co-glycolide) acid (PLGA, 5050) and a blend of PLGA and PHBV. BSA served as a model for hepatocyte growth factor (HGF) since both proteins have similar molecular weights and hydrophilicity. Furthermore, HGF was encapsulated into the PLGA/PHBV composite microsphere with a core-shell structure, and sustained delivery of HGF with maintained bioactivity was achieved for at least 40 days. The moderate degradation rate (about 55% loss of the initial mass) and well-preserved structure after three months of incubation indicated that the PLGA/PHBV composite microspheres would therefore be more suitable than the pure PHBV or PLGA microspheres as a scaffold for engineering liver tissue. PMID- 18431777 TI - The behavior of MC3T3-E1 cells on chitosan/poly-L-lysine composite films: effect of nanotopography, surface chemistry, and wettability. AB - In the present work, a series of composite films were produced from chitosan/poly L-lysine blend solutions. The surface topography, chemistry, and wettability of composite films were characterized by atomic force microscopy (AFM), attenuated total reflection Fourier transform infrared (ATR-FTIR) spectroscopy, and contact angle assay, respectively. For all composite films, blending with poly-L-lysine induced changes in surface chemistry and wettability. Interestingly, it was also found that increasing poly-L-lysine weight fraction in blend solutions could result in different nanoscaled surface topographic features, which displayed particle-, granule-, or fiber-dominant morphologies. MC3T3-E1 osteoblast-like cells were cultured on all composite films to evaluate the effects of surface nanotopography, chemistry, and wettability on cell behavior. The observations indicated that MC3T3-E1 cell behavior was affected by surface topography, chemistry, and wettability simultaneously and that cells showed strong responses to surface topography. On fiber-dominant surface, cells fully spread with obvious cytoskeleton organization and exhibited significantly higher level of adhesion and proliferation compared with particle- or granule-dominant surfaces. Furthermore, fiber-dominant surface also induced greater expression of mature osteogenic marker osteocalcin and higher mineralization based on RT-PCR and von Kossa staining. The results suggest that topographic modification of chitosan substratum at the nanoscale may be exploited in regulating cell behavior for its applications in tissue engineering. PMID- 18431778 TI - SU-8 2000 rendered cytocompatible for neuronal bioMEMS applications. AB - Microfabrication advances have resulted in small, cheap, and precise devices for biological microelectromechanical systems (bioMEMS). SU-8/SU-8 2000 is an attractive material for these applications because of its high-aspect ratio fabrication capability, dielectric properties, and thermochemical stability. Despite these advantages, the potential toxicity of SU-8 2000 may limit its use in cell-based applications. We show that <10% of primary neurons survived when cultured adjacent to or on top of untreated SU-8 2000. We evaluated the efficacy of various detoxification and surface treatments for SU-8 2000 in neuronal cultures after 7-21 days in vitro. Viability was improved to 45.8% +/- 4.5% (mean +/- standard error of the mean) following 3-day heat treatment (150 degrees C) under vacuum, while UV exposure and CO2 supercritical extraction did not improve survival. Furthermore, parylene coating (25 microm), in combination with heat and sonication (in isopropanol) treatments effectively masked the SU-8 2000 and led to 86.4% +/- 1.9% viability. Glow discharge (oxygen plasma) treatment rendered the SU-8 2000 surface more hydrophilic and improved neuronal viability, possibly through improved cell adhesion. No organic leachants were detected by mass spectrometry before or after heat treatment or after sonication. However, XPS analysis revealed the presence of potentially neurotoxic elements, fluorine and antimony. Strategies to improve the cytocompatibility of SU-8 2000 with primary neurons will allow longer culture times and have applications for cell-based microfabrication. PMID- 18431779 TI - In vitro bioactivity of melt-derived glass 46S6 doped with magnesium. AB - Melt-derived glasses in the system SiO(2)-CaO-Na(2)O-P(2)O(5) were synthesized pure or doped with magnesium from 0.4 to 1.2 wt %, for applications as biomaterials in bony surgery. This chemical element has been chosen because of its high physiological interest. Its introduction for different contents in melt derived glasses has never been studied. The bioactivity of glasses was assessed by immersion of the samples in the simulated body fluid solution. Changes in glass surface morphology and composition after immersion were evaluated by several physico-chemical techniques. The aim of this work was to characterize the formation of the apatite-like layer at the glass surface, after in vitro assays and to evaluate the kinetic reaction between the glass and the surrounding synthetic fluids. Results indicate that magnesium influences the formation and the evolution of the newly formed layers: (1) it promotes the dissolution of the silica network, (2) it increases the thickness of the silica gel layer formed conventionally prior to the apatite-like layer, and (3) it slows down the crystallization of the apatite layer. However, the intensity of these effects depends on the content of magnesium introduced in the glass matrix. PMID- 18431780 TI - Polymer surfaces structured with random or aligned electrospun nanofibers to promote the adhesion of blood platelets. AB - Fibrous membranes (nonwoven meshes) prepared via electrospinning technique have great potential in tissue engineering. This work is the first study on the behaviors of blood platelets at the nanostructured surface generated by electrospinning. Poly[acrylonitrile-co-(N-vinyl-2-pyrrolidone)] (PANCNVP) that shows excellent antiplatelet adhesion ability was directly electrospun onto its dense membrane surface. Polyacrylonitrile (PAN) samples were used as controls. The depth as well as the density of the nanofibers can be easily controlled. The results showed that the PANCNVP dense membrane certainly suppressed the activation and adhesion of platelets. However, whether the nanofibers and underlying membranes were composed of PAN or PANCNVP, the nanostructured surfaces promoted the activation, adhesion, and orientation of platelets. It was also found that, if the space between fibers was too large or the depth of fibers was too small, the nanostructured surface did not change the property of antiplatelet adhesion of PANCNVP. The promotion of activation and adhesion of platelets was obviously due to the presence of nanofibers, which induced the changes of surface topography and charge. PMID- 18431781 TI - Improvement of metal and tissue adhesion on surface-modified parylene C. AB - A general method for chemical surface functionalization of parylene C [PC, (para CH2-C6H3Cl-CH2-)n] films is reported. Friedel-Crafts acylation is used to activate the surface of the PC film, and the resulting carbonyl groups are then used to form a range of different organic functional groups to the surface of the parylene film, including alcohol, imine, thiol, phthalimide, amine, and maleimide. The presence of these functional groups on the parylene surface was confirmed by Fourier transform infrared spectroscopy. Static water drop contact angle measurements were also used to demonstrate the changes in hydrophilicity of the PC film surface, consistent with each of the surface modifications. Enhanced metal (gold) adhesion was achieved by anchoring a thiol group onto the acylated surface of PC film. Acylation of parylene with 2-chloropropionyl chloride gave a surface bound chloropropionyl group. Grafting of poly-N-isopropylacrylamide (pNIPAM) onto the chloropropionyl substituted PC film via atom transfer radical polymerization (ATRP) was carried out. The grafted pNIPAM on the parylene surface leads to temperature-dependent cellular tissue adhesion on the PC film. PMID- 18431782 TI - Fabrication of free-standing albumin-nanosheets having heterosurfaces. AB - Sheet-shaped carriers, having both obverse and reverse surfaces and thus a large contact area for targeting a site, have several advantages over spherical-shaped carriers, which have an extremely small contact area for targeting sites. Here, we proposed a novel method to prepare a free-standing ultrathin and biocompatible nanosheet having heterosurfaces, by a combination of four processes: (1) specific adsorption of recombinant human serum albumin (rHSA) molecules onto a patterned octadecyltrimethoxysilane self-assembled monolayer region (ODS-SAM), (2) preparation of nanosheets of rHSA molecules bearing thiol groups (SH-rHSA) via two-dimensionally disulfide crosslinking, (3) surface modification of the resulting nanosheet, and (4) preparation of the free-standing nanosheet by detachment from the ODS-SAM. The SH-rHSA molecules at pH 5.0 and a concentration of 1 microg/mL were specifically adsorbed on the patterned ODS-SAM regions by hydrophobic interaction, and were two-dimensionally crosslinked in the presence of copper ion as an oxidant. The rHSA-nanosheets were then simply detached from the ODS-SAM by treatment with surfactant. We succeeded in the preparation of rectangular (10 microm x 30 microm) and ultrathin (4.5 +/- 1.0 nm) rHSA nanosheets on a patterned ODS-SAM, and could also obtain free-standing rHSA nanosheets having heterosurfaces by surface modification with fluorescent latex beads. Thus, the rHSA-nanosheets having heterosurfaces could be regarded as a new biomaterial for drug carriers, hemostatic reagents, wound dressing for burn injury, and so forth. PMID- 18431783 TI - Immobilization of urease by laser techniques: synthesis and application to urea biosensors. AB - Urease thin films have been immobilized using matrix-assisted pulsed laser evaporation for biosensor applications in clinical diagnostics. The targets exposed to laser radiation were made of frozen composites that had been manufactured by dissolving urease in distilled water. An UV KrF* (lambda = 248 nm, tauFWHM congruent with 30 ns, nu = 10 Hz) excimer source was used for the multipulse laser irradiation of the targets that were cooled down to solidification using Peltier elements. The incident laser fluence was set at 0.4 J/cm2. The surface morphology and chemical bonding states of the laser immobilized urease thin films were investigated by atomic force microscopy and Fourier transform infrared spectroscopy. The enzymatic activity and kinetics of the immobilized urease were assayed by the Worthington method, which monitors urea hydrolysis by coupling ammonia production to a glutamate dehydrogenase reaction. Decreased absorbance was found at 340 nm and correlated with the enzymatic activity of urease. PMID- 18431784 TI - Development of a new poly(ethylene glycol)-graft-poly(D,L-lactic acid) as potential drug carriers. AB - A new poly(ethylene glycol) (PEG)-modified poly(D,L-lactic acid) (PLA) was synthesized by grafting maleic anhydride onto PLA and subsequently amidating with O,O'-bis-(2-aminopropyl) polypropylene glycol-block-polyethylene oxide-block polypropylene glycol (H2N-PEG-NH2, Mw: 600). Its structure was confirmed by FTIR, DSC, 1H NMR, GPC, and ninhydrin test. The polymer is more hydrophilic compared with PLA according to contact angle tests, and is degradable as determined from its pH and mass changes during degradation. The polymer shows a 62.7% decrease in BSA absorption compared with PLA when dried in air, and a 82.76% decrease when dried under 65% humidity, as measured by fluorospectrophotometry. The polymer promotes adhesion and proliferation of osteoblasts, determined by MTT assay. With this new polymer, spherical nanoscale aggregates encapsulated with or without hydrophilic dye are formed spontaneously in water, visualized by inverted microscope and AFM. The particle size is concentration dependent as confirmed by dynamic light scattering, and its critical micelle concentration was 1.124 microg/mL as determined by a fluorescence method. The good hydrophilicity, degradability, cellular compatibility, protein-resistance, self-aggregation, and reactivity of the polymer may lead to its potential applications in drug delivery. PMID- 18431785 TI - Effect of dynamic 3-D culture on proliferation, distribution, and osteogenic differentiation of human mesenchymal stem cells. AB - Ex vivo engineering of autologous bone tissue as an alternative to bone grafting is a major clinical need. In the present study, we evaluated the effect of 3-D dynamic spinner flask culture on the proliferation, distribution, and differentiation of human mesenchymal stem cells (MSCs). Immortalized human MSCs were cultured on porous 75:25 PLGA scaffolds for up to 3 weeks. Dynamically cultured cell/scaffold constructs demonstrated a 20% increase in DNA content (21 days), enhanced ALP specific activity (7 days and 21 days), a more than tenfold higher Ca2+ content (21 days), and significantly increased transcript levels of early osteogenesis markers (e.g., COL1A1, BMP2, RUNX-2) as compared with static culture. Despite the formation of a dense superficial cell layer, markedly increased cell ingrowth was observed by fluorescence microscopy on day 21. Furthermore, increased extracellular matrix deposition was visualized by scanning electron microscopy after 1 and 3 weeks of dynamic culture. The observed increased ingrowth and osteogenic differentiation of 3-D dynamically cultured human MSCs can be explained by generation of fluid shear stress and enhanced mass transport to the interior of the scaffold mimicking the native microenvironment of bone cells. This study provides evidence for the effectiveness of dynamic culture of human MSCs during the initial phase of ex vivo osteogenesis. PMID- 18431786 TI - Electrospinning of multicomponent ultrathin fibrous nonwovens for semi-occlusive wound dressings. AB - This work describes the design and assembly of multifunctional and cost-efficient composite fiber nonwovens as semi-occlusive wound dressings using a simple electrospinning process to incorporate a variety of functional components into an ultrathin fiber. These components include non-hydrophilic poly(L-lactide) (PLLA) as fibrous backbone, hydrophilic poly(vinyl pyrrolidone)-iodine (PVP-I), TiO(2) nanoparticles, zinc chloride as antimicrobial, odor-controlling, and antiphlogistic agents, respectively. The process of synthesis starts with a multicomponent solution of PLLA, PVP, TiO(2) nanoparticles plus zinc chloride, in which TiO(2) nanoparticles are synthesized by in situ hydrolysis of TiO(2) precursors in a PVP solution for the sake of obtaining the particle-uniformly dispersive solution. Subsequent electrospinning generates the corresponding composite fibers. A further iodine vapor treatment to the composite fibers combines iodine with PVP to produce the PVP-I complexes. Experiments indicate that the assembled composite fibers (300-400 nm) possess the ointment-releasing characteristic and the phase-separate, core-sheath structures in which PVP-I residing in fiber surface layer becomes the sheath, and PLLA distributing inside the fiber acts as the core. Based on this design, the structural advantages combining active components endow the assembled composite nonwovens with a variety of functions, especially, the existence of PVP-I, endows the nonwoven with water absorbability, antimicrobial activity, adhesive ability, and transformable characteristic from hydrophilicity to non-hydrophilicity. The multifunctional, cost-efficient, and ointment-releasing characteristics make the multicomponent composite fibrous nonwovens potentially useful in applications such as initial stage of dressing of the cankerous or contaminated wounds. PMID- 18431787 TI - Osteoblast response to continuous phase macroporous scaffolds under static and dynamic culture conditions. AB - Average scaffold pore sizes in the order of several hundred microns are generally required for efficient bone tissue ingrowth in vivo, whereas the culture of large bone engineering constructs in vitro can require bioreactor cultures to decrease diffusional constraints on the cells. In this study, we prepared poly(epsilon caprolactone/D,L-lactide)-based scaffolds with continuous phase macroporosity using a novel CaCl(2) . 6H(2)O porogen agent. Osteogenic differentiation and scaffold colonization in rat bone marrow stromal cell cultures were compared in such polymer scaffolds, and in composites with 30 wt % bioactive glass filler. The effect of a rotating wall bioreactor culture on the cell response was also evaluated. Bioactive filler enhanced proliferation, early osteogenic differentiation, and mineralization of the cultured cells under static conditions. Dynamic cultures, in turn, resulted in decreased cell numbers and inhibition of the differentiation process irrespective of the scaffold type. This effect was ascribed to the harsh mechanical stresses caused by constant collisions of the scaffolds in the bioreactor vessels. However, cells were able to penetrate into the scaffold interior only under dynamic culture conditions. Thus, interconnected macroporosity is an essential, but not sufficient, condition to allow for full colonization of millimeter scale tissue engineering scaffolds in vitro. PMID- 18431788 TI - In vitro and in vivo evaluation of acellular diaphragmatic matrices seeded with muscle precursors cells and coated with VEGF silica gels to repair muscle defect of the diaphragm. AB - In this work, a bioartificial system consisting of VEGF-loaded porous silica gel and myoblasts cultured on acellular diaphragmatic matrix (ADM) has been implanted to repair a surgically created diaphragmatic defect in Lewis rats. ADMs exerted a strong angiogenic response on chorio-allantoic membrane. Cytotoxicity, VEGF release and matrix erodibility in vitro tests demonstrated that the silica support was nontoxic and that the VEGF bioactivity was maintained after matrix entrapment and it was released within a timeframe that can be modulated by synthesis parameters. Different grafts composed by ADMs with and without autologous male myoblasts or/and VEGF-loaded porous silica gel have been implanted to repair previously created diaphragmatic defects in female Lewis rats. Patches composed of ADMs and myoblasts appeared well preserved until 8 weeks, and contained multinucleated cells and cholinergic fibers. At 8 weeks, the implanted cells were still present inside the patches. The disappointing results obtained when VEGF was delivered by porous silica gel were probably due to an abnormal angiogenic response following an excess of local growth factor concentration. Taken together, these results confirmed that our matrices contained biologically active angiogenic factors which were per se sufficient to induce neo-vessels formation, thus allowing the survival of implanted myoblasts. PMID- 18431789 TI - PEOT/PBT based scaffolds with low mechanical properties improve cartilage repair tissue formation in osteochondral defects. AB - The aim of our study was to compare the healing response of biomechanically and biochemically different scaffolds in osteochondral defects created in rabbit medial femoral condyles. A block copolymer comprised of poly(ethylene oxide terephthalate) and poly(butylene terephthalate) was used to prepare porous scaffolds. The 70/30 scaffold (70 wt % poly(ethylene oxide terephthalate)) was compared to the stiffer 55/45 (55 wt % poly(ethylene oxide terephthalate)) scaffold. Nine 6-month-old rabbits were used. Osteochondral defects were filled with 55/45 scaffolds (n = 6); 70/30 scaffolds (n = 6); or left empty (n = 6). Defect sites were allowed to heal for 12 weeks. Condyles were macroscopically evaluated and analysed histologically using the O'Driscoll score for evaluating repair of osteochondral defects. Repair tissue in 70/30 scaffolds consisted of cartilage-like tissue on top of trabecular bone, whereas the tissue within the 55/45 scaffolds consisted predominantly of trabecular bone. O'Driscoll scores for 70/30 scaffolds were significantly better (p = 0.024) in comparison to untreated osteochondral defects and 55/45 scaffolds. This study reveals that the biomechanical and biochemical properties of the scaffold play an important role by themselves, and can affect the healing response of osteochondral defects. Scaffolds with low mechanical properties were superior in cartilage repair tissue formation. PMID- 18431790 TI - Photoinduced in situ formation of various F-actin assemblies with a photoresponsive polycation. AB - We developed a novel in situ method for the control of F-actin assembly by using a synthetic photoresponsive polycation. The photoresponsive polycation mainly comprises a water-soluble cationic monomer and also contains a small amount of the monomer of a triphenylmethane leucohydroxide derivative (20 mol %), which is a well-known photochromic molecule that can be cationized in aqueous solution by ultra violet (UV) irradiation, thereby causing an increase in the total charge on the photoresponsive polycation. Thus, by exposure to UV radiation in aqueous solution, F-actin and the photoresponsive polycation start assembling into F actin/photoresponsive polycation complexes of various morphologies such as bundles, coils, and networks, depending upon the concentrations of both the F actin and salt. Further, localized UV irradiation can be applied in order to control the local formation of F-actin/photoresponsive polycation complexes. Thus, this technique provides a novel method for the spatiotemporal control of F actin assembly and can be applied to investigate the unknown characteristics of F actin. PMID- 18431791 TI - Notice of retraction: Localization of serotonin neuropiles in the brain and thoracic ganglia of the Indian white shrimp, Fenneropenaeus indicus: phylogenetic comparisons and implications for arthropod relationships. PMID- 18431792 TI - Multiparameter detection of apoptosis using red-excitable SYTO probes. AB - Functional assays allowing phenotypic characterization of different cell death parameters at a single-cell level are important tools for preclinical anticancer drug screening. Currently, the selection of cytometric assays is limited by the availability of fluorescent probes with overlapping spectral characteristics. Following on our earlier reports on green and orange fluorescent SYTO probes, we provide herein further insights into applicability of novel red-excitable SYTO stains (SYTO 17, 59-64) for multiparameter analysis of cell fate. In particular, SYTO 62 appears to be a spectrally favorable candidate. Using a correlative comparison between SYTO 16, Annexin V, YO-PRO 1, and fluorescently labeled inhibitors of caspases (FLICA), we demonstrate the specificity of SYTO 62 in detection of apoptotic cell death. Used in conjunction with FLICA or Annexin V, SYTO 62 stain proved amenable for multivariate kinetic analysis of apoptotic events. Considering simplicity of staining protocols, low cost, and avoidance of spectral compensation problems, we expect that red-excitable SYTO dyes will find a wide range of cytometric applications. PMID- 18431793 TI - Standardized method to minimize variability in a functional P2X(7) flow cytometric assay for a multi-center clinical trial. AB - BACKGROUND: Flow cytometric analysis of human P2X(7) pore activity segregates variant from common P2RX7 genotypes and may serve as a biomarker for cancer, pain, inflammation, and immune responses to infection. Standardization is needed to accommodate variable sample age and instrumentation differences in a multicenter clinical trial. METHODS: CD14-PE-stained whole blood samples were treated with YO-PRO-1 combined with a P2X(7) agonist (BzATP) or control, followed by the addition of PI after closure of the P2X(7) pore. Recalled instrument settings from previous publications were used to adapt a standardized fluorescent particle-adjusted set-up method. Experiments were performed to compare the two methods while evaluating components of systematic variability and facilitating reliable processing of samples with varied ages. RESULTS: The median YO-PRO-1 fluorescence of BzATP-treated samples had less variability when collected by the bead-adjusted method and was less influenced by the compensation strategy used. The average day-to-day coefficient of variance for assessments of P2X(7) pore activity by this method was 0.11 +/- 0.04, and the exclusion of nonviable cells was found to accommodate samples aged up to 4 days after phlebotomy. The bead adjusted set-up method produced measurements differing by only 2.0% +/- 1.5% on two analog cytometers, and within similar decades when comparing analog to digital instruments. CONCLUSIONS: These results provide a standardized method for quantitative flow cytometric analysis of P2X(7) receptor phenotypes in blood monocytes with minimal intralaboratory variation and potential for interlaboratory comparisons that can greatly facilitate multicenter functional genomic clinical studies. PMID- 18431794 TI - Study of hydroxyapatite osteoinductivity with an osteogenic differentiation of mesenchymal stem cells. AB - Osteoinductivity of hydroxyapatite (HA) was investigated using uncommitted pluripotent mouse stem cells, C3H10T1/2 in an in vitro differentiation assay. For comparative analysis, the cells were cultured on substrates made of osteoinductive HA, with biocompatible titanium and plastics as the negative control. HA exhibited the ability to induce expression of osteo-specific genes in C3H10T1/2, including alkaline phosphatase (ALP), type I collagen, and osteocalcin; compared with its insignificant up-regulation of the same genes in osteoblast-like cells, Saos-2. HA osteoinductivity exhibited in C3H10T1/2 was comparable to that of a bone morphogenetic protein (BMP) with reference to the up regulation of osteo-specific genes except the core binding factor 1 (Cbfa1, Runx). This result implies a difference in osteogenic induction pathway initiated by HA and BMP. Using this mesenchymal stem cells (MSC) culture assay, osteoinductivity was also demonstrated to be present in the conditioned medium derived from MSC cultured on HA substrates. This conditioned medium exhibited excellent ability to up-regulate ALP in the absence of HA and BMP. The results suggest that the HA can interact with the cells and generate potent inductive substance released into the medium. Such substance in turn is able to induce uncommitted cells to differentiate into the osteolineage. PMID- 18431795 TI - A rapid flow cytometry test based on histone H2AX phosphorylation for the sensitive and specific diagnosis of ataxia telangiectasia. AB - Ataxia telangiectasia (A-T) is a progressive neurodegenerative disease with onset in early childhood, caused by mutations in the ATM (ataxia-telangiectasia mutated) gene. Diagnosis relies on laboratory tests showing high levels of serum alphafetoprotein, cell sensitivity to ionizing radiation (IR) and absence or reduced levels of ATM protein. Many tests, however, are not sufficiently sensitive or specific for A-T, have long turnaround times, or require large blood samples. This prompted us to develop a new flow cytometry method for the diagnosis of A-T based on the measurement of histone H2AX phosphorylation. We established normal ranges of histone H2AX phosphorylation after 2 Gy IR by testing T-cell lines, lymphoblastoid cell lines (LCLs) and/or peripheral blood mononuclear cells (PBMCs) or both from 20 genetically proven A-T and 46 control donors. To further evaluate the specificity and sensitivity of the test, we analyzed cells from 19 patients suspected of having A-T, and from one Friedreich Ataxia, one Ataxia with Oculomotor Apraxia type 2, and one Nijmegen Breakage Syndrome patients. Phosphorylated histone H2AX mean fluorescence intensity of irradiated A-T cells was significantly lower than that of healthy donors. The intrastaining, intraassay, and interassay imprecisions were 85th%). CONCLUSION: Results suggest that maternal depressive symptoms are related to overweight in children aged 6-24 months. PMID- 18431838 TI - Association between annoyance and individuals' values of nitrogen dioxide in a European setting. AB - INTRODUCTION: Annoyance caused by air pollution has been proposed as an indicator of exposure to air pollution. The aim of this study was to assess the geographical homogeneity of the relationship between annoyance and modelled home based nitrogen dioxide (NO2) measurements. METHODS: The European Community Respiratory Health Survey II was conducted in 2000-1, in 25 European centres in 12 countries. This analysis included 4753 subjects (from 37 in Tartu, Estonia, to 532 in Antwerp, Belgium) who answered the annoyance question and with available outdoor residential NO2 (4399 modelled and 354 measured) including 20 cities from 10 countries. Annoyance as a result of air pollution was self-reported on an 11 point scale (0, no disturbance at all; 10, intolerable disturbance). Demographic and socioeconomic factors, smoking status and the presence of respiratory symptoms or disease were measured through a standard questionnaire. Negative binomial regression was used. RESULTS: The median NO2 concentration was 27 microg.m(-3) (from 10 in Umea, Sweden, to 57 in Barcelona, Spain). The mean of annoyance was 2.5 (from 0.7 in Reykjavik, Iceland, to 4.4 in Huelva, Spain). NO2 was associated with annoyance (ratio of the mean score 1.26 per 10 microg.m(-3), 95% CI 1.19 to 1.34). The association between NO2 and annoyance was heterogeneous among cities (p for heterogeneity <0.001). CONCLUSIONS: Annoyance is associated with home outdoor air pollution but with a different strength by city. This indicates that annoyance is not a valid surrogate for air pollution exposure. Nevertheless, it may be a useful measure of perceived ambient air quality and could be considered a complementary tool for health surveillance. PMID- 18431840 TI - Proceedings of the International Conference: Advances in Pneumology, 10-12 May 2007, Wuppertal, Germany. PMID- 18431839 TI - Safety of very tight glucose control in type 2 diabetes in question. PMID- 18431841 TI - Symposium in tribute to Dr. Anthony F. DePalma, first editor-in-chief of Clinical Orthopaedics and Related Research. PMID- 18431842 TI - Proceedings of the 8th Fatty Acid and Cell Signaling (FACS) Workshop, June 26-28, 2007, Quebec City, Canada. PMID- 18431844 TI - Health care needs heart. PMID- 18431843 TI - The c-Jun N-terminal kinase JNK functions upstream of Aurora B to promote entry into mitosis. AB - Mitogen-activated protein kinases (MAPKs) are components of signaling cascades regulated by environmental stimuli. In addition to participating in the stress response, the MAPKs c-Jun N-terminal Kinases JNK1 and JNK2 regulate the proliferation of normal and neoplastic cells. JNKs contribute to these processes largely by phosphorylating c-Jun and thus contributing to the activation of the AP-1 complex. We here report that JNKs control entry into mitosis. We have observed that JNK activity and phosphorylation of c-Jun become elevated during the G2/M transition of the cell cycle in immortalized fibroblasts and ovarian granulosa cells. Pharmacological inhibition of JNK causes a profound cell cycle arrest at the G2/M transition in both cell types. This effect is specific as it occurs with two distinct small molecule compounds. Inactivation of JNK prior to mitosis prevents expression of Aurora B and phosphorylation of Histone-H3 at Ser 10. Silencing of JNK1 and 2 causes a similar effect, whereas overexpression of JNK1 and 2 causes the opposite effect. Inhibition of JNK delays activation of cdc 2 and prevents downregulation of Cyclin B1. We conclude that JNK signaling promotes entry into mitosis by promoting expression of Aurora B and thereby phosphorylation of Histone-H3. PMID- 18431845 TI - Patient satisfaction. CMS set to release new patient satisfaction results. Are you ready? PMID- 18431846 TI - Too harsh. PMID- 18431847 TI - On seeking recognition and fair pay. PMID- 18431848 TI - Numerical human models for accident research and safety - potentials and limitations. AB - The method of numerical simulation is frequently used in the area of automotive safety. Recently, numerical models of the human body have been developed for the numerical simulation of occupants. Different approaches in modelling the human body have been used: the finite-element and the multibody technique. Numerical human models representing the two modelling approaches are introduced and the potentials and limitations of these models are discussed. PMID- 18431849 TI - Human cortical bone: the SINUPROS model. AB - Several modelizations have been investigated on human cortical bone in our team and we have often observed that the introduction of a new geometrical parameter induces significant perturbations on the numerical values obtained with previous models. We have therefore decided to take into account the totality of all possible parameters in a modelization which is physically and physiologically plausible. In order to do this, we have analyzed the architecture of cortical bone and exhibited all parameters that occur. To determine physical properties at each architectural level, the best adapted tool is without any doubt the mathematical theory of homogenization. All the necessary algorithms have been implemented into SINUPROS software (websites of the Universities). Its main interest is the evaluation of macroscopic physical properties for a given configuration. It can also be used to seek, by successive tests, configurations corresponding to properties experimentally measured. Computation time being too high (10 to 45 minutes according to tested configurations), a fast version based on approximation theory has been developed and thus the obtaining of the results is immediate. The researched configuration being thus obtained, it has then to be validated by the original version. PMID- 18431850 TI - You've got mail: erj@lumc.nl. PMID- 18431851 TI - Rapid, reliable and responsive: for whom? PMID- 18431852 TI - Unstable operators in image processing. AB - Digital image processing is a powerful tool in biomedical applications and is used for image enhancement, segmentation or classification. Unfortunately, some of the processes in image analysis are unstable operators. Even small perturbations in the image data can cause significant errors in the result. The corresponding practical algorithms are sensitive to deviations in the input data and the behaviour is similar to linear systems with ill conditioned matrices. This property is related to the so called ill posed problems. We review the framework of ill posed problems and regularization in image analysis. We examine a few examples related to biomedical imaging: segmentation by thresholding, edge detection, length estimation, orientation. PMID- 18431853 TI - Bone as a structural material: how good is it? AB - As a structural material, bone is not very good; compared to engineering materials such as metal alloys and fibre composites, its mechanical properties are mediocre. In fact, the really amazing thing about bone is that it is able to achieve even these mediocre properties with the ingredients available: hydroxyapatite, collagen and water. Drawing on previous research, and some simple fracture mechanics calculations, we can see how bone optimises the use of these materials in a composite structure which has important features at two different scales: the nanometre scale and the hundred-micron scale. PMID- 18431854 TI - [Perceptions of the Swedish method and the consequences for the development of gymnastics in France before 1914]. PMID- 18431855 TI - Bone strength in pure bending: bearing of geometric and material properties. AB - Osteoporosis is characterized by decreasing of bone mass and bone strength with advanced age. For characterization of material properties of dense and cellular bone the volumetric bone mineral density (vBMD) is one of the most important contributing factors to bone strength. Often bending tests of whole bone are used to get information about the state of osteoporosis. In a first step, different types of cellular structures are considered to characterize vBMD and its influence to elastic and plastic material properties. Afterwards, the classical theory of plastic bending is used to describe the non-linear moment-curvature relation of a whole bone. For bending of whole bone with sandwich structure an effective second moment of area can be defined. The shape factor as a pure geometrical value is considered to define bone strength. This factor is discussed for a bone with circular cross section and different thickness of cortical bone. The deduced relations and the decrease of material properties are used to demonstrate the influence of osteoporosis to bone bending strength. It can be shown that the elastic and plastic material properties of bone are related to a relative bone mineral density. Starting from an elastic-plastic bone behavior with an constant yield stress the non-linear moment-curvature relation in bending is related to yielding of the fibres in the cross section. The ultimate moment is characterized by a shape factor depending on the geometry of the cross section and on the change of cortical thickness. PMID- 18431856 TI - [Catholic sport in France at the beginning of the 20th century]. PMID- 18431857 TI - Bone quality issues an matrix properties in OP cancellous bone. AB - There is increasing evidence nowadays that diseases or conditions, like osteoporosis (OP), which are conventionally defined in terms of bone quantity/mass, are also associated with concomitant changes at the bone matrix level. The present study examined the composition, density and mineral content of OP cancellous bone at the tissue level and the hardness values at the trabecular level to establish correlations between these variables. The results showed that changes in porosity (Bone volume/Tissue volume) are accompanied by changes in mineral content and in the hardness of individual trabeculae. In other words in OP there are both quantitative and qualitative effects that take place with the progress of this condition. PMID- 18431858 TI - English influences on French sport: "anglomania" and national revival, 1870-1914. PMID- 18431859 TI - Colorectal polyps: clinical significance of endoscopic and pathologic correlation. PMID- 18431860 TI - Invalid comparison of the IMMY Histoplasma antigen assay with the "gold standard". PMID- 18431861 TI - WITHDRAWN: Irinotecan plus weekly 5-fluorouracil and leucovorin as salvage treatment for patients with metastatic colorectal cancer: a phase II trial. Dig Dis 2007;25:100-5. PMID- 18431862 TI - Telerehabilitation: current perspectives. AB - Telerehabilitation in which rehabilitation services are provided at a distance using communication technologies is a new and developing field of telehealth. Primarily developed to provide equitable access to individuals who are geographically remote and to those who are physically and economically disadvantaged, telerehabilitation also has the capacity to improve the quality of rehabilitation health care. Online delivery of rehabilitation enables the rehabilitation therapist to optimize the timing, intensity and duration of therapy that is often not possible within the constraints of face-to-face treatment protocols in current health systems. This chapter outlines the advances made to date in telerehabilitation applications in the fields of physiotherapy, speech-language pathology, occupational therapy, and biomedical engineering and provides evidence for the success of these applications. Applications to date encompass systems ranging from low-bandwidth low-cost videophones, to highly expensive, fully immersive virtual reality systems with haptic interfaces. A number of barriers to the establishment and advancement of telerehabilitation within health care systems have been outlined and include professional issues relating to the inherent hands-on approach of some treatments, licensure laws, professional skill development, patient disability, reimbursement, and the paucity of online assessment and treatment tools and outcomes data. In response, possible solutions to these barriers such as the development and validation of alternative assessment and treatment procedures, involvement in the international policy debate, as well as the resolution of national professional policies which hinder the wider uptake of telerehabilitation technologies, have been outlined. The future of telerehabilitation is promising as a new, yet complex form of telehealth with the capacity to provide a wide range of services specifically designed to suit the needs of the individual. PMID- 18431863 TI - [Significance of magnetic resonance tomography and electronic axiography in diagnostics of TMJ dysfunctions]. PMID- 18431864 TI - [Clinical aspects of microbial colonization of temporary dentures from acrylates]. PMID- 18431865 TI - [Marginal seating of cast crowns depending on luting cements]. PMID- 18431866 TI - [Algorithm of children cleft lip and palate treatment standardization with use of the computer methods of data record]. PMID- 18431867 TI - [Indices of stomatological status in the staff of factories with radiation: dangerous conditions of work]. PMID- 18431868 TI - [Tooth preparation peculiarities for reimplantation in case of its full dislocation]. PMID- 18431869 TI - [Technology of stomatologic organization Web-site development or interactive marketing]. PMID- 18431870 TI - [Role of psychological and social factors in forming in doctors-stomatologists the syndrome of emotional burnout]. PMID- 18431871 TI - [Methodology of new pharmacoeconomic method "impact on hospital budget" by the example of local hemostatic agents in abdominal surgery]. PMID- 18431872 TI - [Local form of autoimmune pancreatitis]. PMID- 18431873 TI - [Posttraumatic hematoma simulating abdominal tumor]. PMID- 18431875 TI - [Methods of treatment of acute pleural empyema]. PMID- 18431874 TI - [Trombendarterectomia from three arterial beds at critical ischemia]. PMID- 18431876 TI - Medicine meets engineering. Preface. PMID- 18431877 TI - [Problems of ethiopatogenesis of acute gastroduodenal ulcers at urgent conditions]. PMID- 18431878 TI - [Antisepsis and J. Lister. To 140th anniversary of method and 180th birthday anniversary of author]. PMID- 18431879 TI - Insights about nuclear medicine technology: findings from a 2006 survey of NMT education program directors. PMID- 18431880 TI - Protecting and promoting the rights and welfare of students. Interview by Teresa O'Connor. PMID- 18431882 TI - Implications of diabetes mellitus in patients with STEMI: data from Thai ACS Registry. AB - BACKGROUND: Data exist on the community-based perspective on the relation of diabetes mellitus (DM) and acute ST elevation myocardial infarction (STEMI) worldwide but no data is available in Thailand. MATERIAL AND METHOD: The Thai Acute Coronary Syndrome Registry (TACSR) is an observational study of patients hospitalized with ACS at 17 hospitals in different regions of Thailand. The present sub-study sample consisted of 3,725 patients with STEMI in a 3-year period. RESULTS: Nearly 40% (37.15%) of them had DM who were older predominantly women, with a greater prevalence of co-morbidities and fewer current smokers. Patients with DM who developed STEMI were at increased risk for heart failure, arrhythmia, bleeding and death. These differences remained after adjustment for potential confounding prognostic factors. CONCLUSION: A considerable proportion of patients with STEMI have DM. This proportion is higher than any other studies published. Diabetic patients who developed STEMI are also at increased risk for adverse outcomes including, heart failure, arrhythmia, bleeding and death compared to patients without DM. PMID- 18431881 TI - Demographic, management practices and in-hospital outcomes of Thai Acute Coronary Syndrome Registry (TACSR): the difference from the Western world. AB - BACKGROUND: To establish a national registration of acute coronary syndrome (ACS) registry in Thailand by networking health service institutions to determine the demographic, management practices, and in-hospital outcomes of patients with ACS. MATERIAL AND METHOD: The Thai ACS registry is a multi-center prospective project of nationwide registration in Thailand. Institutions were invited to participate in the registry through members of the Heart Association of Thailand. A series of workshops were organized to ensure standardization and quality control of the data and conduct of the present study. Web-based double data entry was used and the data were centrally managed and analyzed. RESULTS: The enrollment of the patients started in August 2002. After three years, records of 9,373 patients were collected from 17 hospitals. The patients were classified as ST elevation myocardial infarction (STEMI) (40.9.%), non-ST-elevation myocardial infarction (NSTEMI) (37.9%) and unstable angina (UA) (21.2%). The STEMI group was younger predominantly male, with a fewer number of diabetes than NSTEMI or UA. About half of the STEMI patients (52.6%) received reperfusion therapy. Primary percutaneous coronary intervention (PCI) was performed in 22.2% of STEMI. The median door to needle and door to balloon time were 85.0 and 122 minutes respectively. The median times to treatment were 240 minutes in the thrombolysis group and 359 minutes in the primary PCI group. Nearly half of NSTEMI and UA went to coronary angiography and about one-fourth of them received revascularization either PCI or coronary artery bypass grafting in the same admission. The total mortality rate was high in STEMI (17.0%) followed by NSTEMI (13.1%) and UA (3.0%). CONCLUSION: Thai ACS registry provides a detail of demographic, management practices, and in hospital outcomes of patients with ACS. Time from onset to admission, door to needle time and door to balloon time were considered as suboptimal. Overall, in hospital mortality is higher than reports from Western countries. The raising awareness among the general population about urgency of seeking medical attention for chest pain and concerted effect to improve in-hospital time delay is warranted. These data may have an impact on our health care system and alert the government to adopt an appropriate policy to solve these problems. PMID- 18431883 TI - Costs of payment in Thai acute coronary syndrome patients. AB - BACKGROUND: Acute coronary syndrome (ACS) is a major health care syndrome that can financially burden patients throughout the world, including Thailand. Few studies purposed estimating the costs of treatment. The data from the ACS registry database represented the costs of hospital charges paid by ACS patients. Although these were not the actual treatment costs, the authors can approximately estimate the total expenditure for the first admission. OBJECTIVES: First, calculate the cost of ACS to the patients, including diagnostic, demographic data, treatment modalities, type of payers, hospital profile, and outcomes. Second, find the appropriate model to identify the independent factors for predicting the treatment costs. MATERIAL AND METHOD: The present study collected data from the second and third phase of a national multicenter prospective registry of ACS in Thailand, Thai ACS registry (TACSR). 3,552 patients with new onset of ACS were analyzed. RESULTS: Median age was 67 years (range 26.5-105.5) with predominately male and median length of stay (LOS) was 7 days (range, 1 184). 42% referred from other hospitals. The median cost of the total population was 47,908 baht (range, 633-1,279,679). When classified into those of STEMI, NSTEMI, and UA, the costs were 82,848.5, 40,531 and 26,116 baht respectively, p < 0.0001. Patients in the government hospital had to pay the total cost with PCI and CABG, 152,081-161,374 baht and 203,139-223,747 baht respectively, while the private hospital charged almost twice as much. For the types of payers, private insurance including private employee security fund paid significantly more than others. Costs in patients paid by "30 baht na tional health scheme and social security fund" were significantly less than those of others. For modality of treatment in STEMI, primary PCI was significantly more costly than thrombolytics and no reperfusion therapy, 161,096.5 vs. 60,043.0 and 33,335.0 baht respectively p < 0.0001. Early invasive groups in NSTEMI/UA had much higher median costs 145,794.0 baht when compared to those of the conservative group, 47,908 baht, p < 0.0001. Two multiple linear regression models according to the diagnostic group identified the independent factors for predicting cost. PCI, LOS, CABG, admission in a private hospital, Death, GPIIb/IlIa inhibitors use, major bleeding, coronary angiogram, thrombolytics use, age and diabetes were independent predictors for the cost in STEMI patients, R2 = 0.58. For those of NSTEMI/UA, the independent predictors for the cost were PCI, LOS, CABG, admission in a private hospital, death, GP IIb/IIIa inhibitors use, major bleeding, coronary angiogram, age, ventricular arrhythmia, CHF and referred patients, R2 =0.62. CONCLUSION: Costs in ACS patients were markedly different among diagnostic groups. The clinical risk factors were hospital type, type of payers, referred system, treatment procedures, drugs used and complications including outcome. Some of these factors could independently predict the costs. PMID- 18431884 TI - Renal dysfunction as an independent predictor of total mortality after acute coronary syndrome: the Thai ACS Registry. AB - BACKGROUND: Renal insufficiency in the acute coronary syndrome (ACS) is associated with poor cardiac outcome. In Asian populations, there are no data available for these associations. MATERIAL AND METHOD: Data was from the Thai ACS registry, only a new case of ACS. Clinical characteristics, treatment strategies, in-hospital mortality and 1-year mortality were compared for patients with normal or mild renal dysfunction (estimated glomerular filtration rate [eGFR]> 60 ml/minute/1.73 m2, n = 809 [44.5%]), moderate renal dysfunction (eGFR 30-60 ml/minute/1.73 m2, n = 706 [38.9%]), and severe renal dysfunction (eGFR < 30 ml/minute/1.73 m2, n = 301 [16.6%]). RESULTS: Of the 1,816patients with mean follow-up 10.8 months, the mean age was 65 years, and 59.2 percent of the groups were male. Patients with severe renal dysfunction were significantly older, less likely to be male (45.2%, p < 0.001) and had a greater prevalence of diabetes (63.1%, p < 0.001) and hypertension (85.4%, p < 0.001). In-hospital and 1-year mortality were 13.5% and 22.5% respectively. According to discharge diagnosis, unadjusted hazard ratios for overall in-hospital mortality was statistically significant only in ST elevation MI subgroup, hazard ratio was 2.73 (95% CI, 1.72 to 4.34) and 6.27 (95% CI, 3.78 to 10.4) for moderate and severe renal dysfunction group, respectively. The risk of death for all types of ACS at 1-year follow up increased when eGFR decreased below 60 ml/minute/1.73 m2, the adjusted hazard ratio was 1.66 (95% CI,1.22 to 2.23) and 1.91 (95% CI, 1.34 to 2.72) for moderate and severe renal dysfunction group, respectively. CONCLUSION: From Thai ACS registry, renal dysfunction at presentation is an independent predictor for the overall 1-year mortality and appeared to associate with an increase in hospital mortality in the subsets with STEMI PMID- 18431885 TI - Predictors of in-hospital mortality in non-ST elevation acute coronary syndrome in Thai Acute Coronary Syndrome Registry (TACSR). AB - OBJECTIVE: To determine baseline prognostic factors of in-hospital mortality in Thai patients with non-ST-elevation acute coronary syndrome (NSTE-ACS). MATERIAL AND METHOD: Among 5,537 NSTE-ACS patients enrolled in Thai Acute Coronary Syndrome Registry, a univariate analysis and multivariate analysis were used to estimate the relationship of baseline clinical variables and in-hospital mortality. Variables examined included demographics, history and presenting characteristics. RESULTS: The in-hospital mortality rate was 9.5%. The statistically significant, adjusted baseline prognostic factors of in-hospital death were older age > or =65 years) (odds ratio [OR] 2.2, 95% confidence interval [CI] = 1.54-3.09), shock at presentation (OR 4.6, 95%CI = 2.91-7.32), heart failure (OR 3.1, 95%CI = 2.15-4.38), positive cardiac marker (OR 1.7, 95%CI = 1.18-2.53), arrhythmia (OR 12.3, 95%CI = 8.71-17.35), major bleeding (OR 2.9, 95%CI = 1.84-4.51), and cerebrovascular accident (OR 4.9, 95% CI = 2.42-9.97). While dyslipidemia (OR 0.6, 95%CI = 0.45-0.87), having percutaneous coronary intervention (OR 0.6, 95% CI = 0.39-0.94), receiving aspirin (OR 0.6, 95%CI = 0.33-0.94), beta-blocker (OR 0.5, 95% CI = 0.40-0.73), angiotensin converting enzyme inhibitor (OR 0.6, 95% CI = 0.43-0.78) and nitrate (OR 0.5, 95%CI= 0.35 0.76) were associated with lower in-hospital mortality. CONCLUSION: The in hospital mortality is higher in Thai NSTE-ACS patients compared to other populations. The present study supports and confirms the prognostics importance of several baseline characteristics reported in previous studies. PMID- 18431886 TI - Diabetes mellitus and non-ST elevation myocardial infarction in Thai ACS Registry. AB - OBJECTIVE: To describe differences in in-hospital morbidity and mortality, presenting characteristics and management practices of diabetic and non-diabetic patients with non-ST elevation myocardial infarction using data from Thai ACS registry. MATERIAL AND METHOD: Thai ACS registry is a multi-center prospective project of nationwide registration in Thailand. RESULTS: The present study consisted of 3,548 patients with non-ST elevation myocardial infarction from 17 hospitals in about a 3-year period. About 50% of the patients with diabetes were more often female, with a greater prevalence of hypertension and dyslipidemia. The diabetic group was at an increased risk for congestive heart failure (adjusted odds ratio 1.84) but not increased risk for cardiac arrhythmia, cardiac mortality, and in-hospital mortality. CONCLUSION: There was a very high prevalence of diabetes in non-ST elevation myocardial infarction from ThaiACS registry. These patients were at increased risk for congestive heart failure as index of hospitalization but were not at increased risk for in-hospital mortality when compared with patients without diabetes. PMID- 18431887 TI - Cardiac arrhythmias in Thai Acute Coronary Syndrome Registry. AB - BACKGROUND: The incidence of arrhythmic complications in Thai patients with acute coronary syndromes (ACS) has not been previously reported. The present study results will serve as the local database for future studies. OBJECTIVE: To evaluate the incidence of arrhythmic complications in ASC in Thai patients and to identify factors that may affect arrhythmia complications in ACS patients. MATERIAL AND METHOD: Data collected from 9,373 patients from the Thai acute coronary syndrome registry (TACSR) were analyzed. This registry includes patients who presented with ACS including ST elevation myocardial infarction (STEMI), non ST elevation myocardial infarction (NSTEMI) and unstable angina (UA), within 14 days from the symptoms onset. RESULTS: 395 (4.2%) patients with an ACS presented after cardiac arrest. These patients were noted to have significantly higher in hospital mortality (50.1%). The incidence of serious cardiac arrhythmia complications in the TACSR was 16.6%. Among them, 62.7% were sustained VT/VE 31.5% had second or third degree AV block, and 5.8% has both VT/VF and AV Block. The incidence of VT was higher in the younger age group, while AV block and arrhythmic death were higher in the older aged patients. Arrhythmias complicating ACS were associated with increased mortality risk. Congestive heart failure (CHF) within the first 48 hours, current use of tobacco and cardiac troponin elevation were associated with significantly higher arrhythmic complications during hospitalization. CONCLUSION: Arrhythmias complicating ACS were associated with higher in hospital mortality. CHF within the first 48 hr, current tobacco use and cardiac troponin elevation were associated with significantly higher arrhythmic complications. PMID- 18431888 TI - Impact of gender on treatment and clinical outcomes in acute ST elevation myocardial infarction patients in Thailand. AB - BACKGROUND: Coronary heart disease is the leading cause of mortality and morbidity in men as well as in women. Women have their first cardiac event 6 to 10 years later than men do. Whereas, the cardiovascular death rates are declining in men, they remain constant in women. In cardiovascular studies with age limits, women are naturally the minority, amounting to < 40%. OBJECTIVE: Determine the effect of gender on treatment and clinical outcomes in acute ST elevation myocardial infarction (STEMI) patients. MATERIAL AND METHOD: This is a multicenter study including 13 government and 4 private institutions. Between August 1, 2002 and October 31, 2005, 3,836 consecutive patients with ST elevation myocardial infarction were enrolled. The patient characteristics, treatment, and hospital outcome were collected and validated. In-hospital management and outcomes were compared between men and women, without adjustment, with adjustment for age alone and with adjustment for age and other covariates by means of multivariate stepwise logistic regression analysis. In each model tested, gender was forced into the model, whereas other predictors were selected in a stepwise manner RESULTS: Women were 1,223 patients (31.9%) of all patients and were 8 years older than men (67.5 +/- 12.0 vs. 59.7 +/- 12.4 years). Women had a higher incidence of diabetes and hypertension (46.9% vs. 31.0%, p < 0.001 and 62.1% vs. 45.3%, p < 0.001). Smoking and family risk factor were found in men more than in women (17.5% vs. 52.5%, p < 0.001 and 7.4% vs. 11.2%, p< 0.001). Women presented more frequently with cardiac dyspnea and shock than men (35.3% vs. 22.2%, p < 0.001 and 21.5% vs. 13.8%, p < 0.001). There was no difference in time to admission between men and women. Beta blocker, statin, angiotensin converting enzyme inhibitor (ACEI) and angiotensin II receptor inhibitor (ARB) were less frequently used in women. Coronary angiogram was performed less often in women and less abnormal angiogram were found (57.8% vs. 65.0%, p < 0.001 and 55.5% vs. 63.9%, p < 0.001). A lower percentage of women received thrombolytic therapy (28.1% vs. 31.4%, p = 0.039) as well as percutaneous coronary intervention (PCI) (39.0% vs. 48.5%, p < 0.001).Unadjusted in-hospital mortality was significantly higher in women than in men (23.6% vs. 13.9%, unadjusted OR (95% CI) 1.90 (1.60 2.26), p < 0.001). After adjustment of other covariables (age group> 75 years, hospital group (metropolitan, regional), chest pain, cardiac dyspnea, shock, post cardiac arrest, diabetes, hypertension, family history, smoking, dyslipidemia, refer time to admission < or = 12 hours (hour), aspirin, beta blocker calcium antagonist, statin, ACEI, ARB, nitrate, coronary angiogram, thrombolysis, PCI, coronary artery bypass grafting, congestive heart failure, arrhythmia, stroke, bleeding), the gender difference in-hospital mortality no longer existed (adjusted OR (95% CI) 1.03 (0.80-1.33), p = 0.814). The incidence of congestive heart failure as a complication was significantly higher in women than in men (55.6% vs. 38.7%, p < 0.001, adjusted OR (95% CI) 1.29 (1.08-1.54), p = 0.005). CONCLUSION: Women with acute ST elevation myocardial infarction were older and had a higher incidence of hypertension and diabetes than men. Women were less likely than men to undergo coronary angioplasty. Women in the authors' registry had a higher risk for in-hospital morbidity and mortality than men. Early and aggressive treatment might improve the clinical outcomes in women with STEMI. PMID- 18431889 TI - In-hospital invasive strategy and outcomes in non-ST elevation acute coronary syndrome management from Thai Acute Coronary Syndrome Registry. AB - BACKGROUND: The management guideline of acute coronary syndrome has been changed in recent years, especially in the group of non-ST elevation acute coronary syndrome (NSTE-ACS). Presently, there is no existing guideline in Thailand. Additionally, several different management factors of health care from Western countries being used. OBJECTIVE: Evaluate the real management strategy, including the utilization of invasive management and pharmacotherapy of NSTE-ACS Thai patients in the absence of official clinical management guideline. MATERIAL AND METHOD: Thai Acute Coronary Syndrome (Thai ACS) Registry is a large, observational prospective, population-based registry designed to collect the data of "real-life" patient management. The full details of methods have been published in the present supplement. RESULTS: Five thousand five hundred and thirty-seven consecutive patients were registered between August 1, 2002 and July 31, 2005. Among these patients, 3,548 (64.1%) were diagnosed with non-ST elevation myocardial infarction (NSTEMI) and 1,989 (35.9%) with unstable angina. Coronary angiography was performed during hospitalization in 2,476 patients (44.7%). From 2,476 patients who underwent coronary angiogram, 405 (16.4%) had revascularization within 2 days and 1,019 (42.9%) after 2 days. Overall, in hospital mortality was 9.5% and cardiac mortality was 6.3%. Patients with NSTEMI had more than 4 times the mortality of patients with unstable angina (13.1 vs. 3.0%, p < 0.001). Patients who received only medical treatment without coronary angiogram had the highest mortality rate. The in-hospital outcomes were not different between patients who received early or delayed revascularization. CONCLUSION: Mortality rate of NSTEACS in the authors' registry was very high. NSTEMI had the worse prognosis. Invasive strategy is associated with better in hospital outcome but is underutilized. PMID- 18431891 TI - Predictors of in-hospital mortality in Thai STEMI patients: results from TACSR. AB - BACKGROUND: Clinical predictors of high-risk STEMI patients may guide physicians to the type of treatment, as high-risk patients need more aggressive treatment than low-risk patients. There was no previous registry of STEMI patients in Thailand. OBJECTIVE: To determine the clinical predictors of in-hospital mortality in STEMI patients from the Thai ACS MATERIAL AND METHOD: A multi-center prospective nationwide Thai Acute Coronary Syndrome Registry (TACSR) was done between August 1, 2002 and October 31, 2005. The STEMI patients were registered to Thai ACS web site. Clinical and demographic characteristics, coronary risk factors, presenting symptoms, in-hospital treatments, reperfusion procedures and the patients' outcomes were recorded and analyzed. RESULTS: 3,836 STEMI patients were studied. The mean age was 62.2 +/- 12.8 years and 68% of the patients were male. The mortality rate of Thai STEMI patients was 17% (86.8% from cardiac causes) and the main cause of death was pumping failure (61.3%). The patients with older age > or = 75 years, patients with diabetes, shock, and cardiac arrhythmias had a higher mortality (29.4, 21.2, 43.4 and 37.24% respectively), while patients who underwent primary percutaneous coronary intervention (primary PCI) had a lower mortality rate (12.66%). Patients who received treatment with ASA, beta-blocker ACE inhibitor/ARB and statin had lower in-hospital mortality. CONCLUSION: The clinical predictors of high in-hospital mortality in STEMI patients from the TACSR were older age > or = 75 years, diabetes, shock, and cardiac arrhythmias. The treatments that can decrease the mortality were primary PCI, ASA, beta-blocker, ACE inhibitor/ARB and statin. PMID- 18431890 TI - Acute coronary syndrome in young adults: the Thai ACS Registry. AB - BACKGROUND: There are few data regarding acute coronary syndrome (ACS) in young adults. ACS in young adults may have some characteristics that are different from those in older patients. OBJECTIVE: The purpose of the present study was to assess the frequency, risk factors, presenting symptoms, treatment, complications and in-hospital outcomes of young patients with ACS in Thailand compared with those of older patients. MATERIAL AND METHOD: From the Thai ACS registry database of 9,373 consecutive patients admitted to participating hospitals between August 1, 2002 and October 31, 2005, the authors divided patients into three age categories: < 45 years, 45-54 years and > 54 years. Risk factors, presenting symptoms, type ofACS, management, complications and in-hospital outcomes of the 3 age groups were analyzed. RESULTS: Young patients comprised of 5.8% (544 patients) of all ACS patients. Discharge diagnosis in the young group was ST segment elevation myocardial infarction (STEMI) in 67%, non-ST segment elevation myocardial infarction (NSTEMI) 20% and unstable angina 14%. The young patients were more likely to have an STEMI than their elder counterparts. Risk factors such as tobacco use and a family history were more frequent in the young patients, whereas diabetes and hypertension were less frequent. Importantly, 66% of the patients aged <45 years had a history of tobacco use. A higher percentage of the young patients underwent coronary angiography, percutaneous coronary intervention and received aspirin, thienopyridines, GP IIb/ IIIa antagonists, beta-blockers and statins. In STEMI patients, reperfusion therapy was given more frequently in the patients aged < 45 years. Younger patients had a lower in hospital mortality rate, lower incidence of congestive heart failure and a shorter length of stay. Multivariable analysis of in-hospital mortality revealed that older age remained an independent predictor of death. CONCLUSION: In Thailand, 5.8% ofpatients with ACS are under the age of 45 years old. The frequency of risk factors in the young patients differs from those in their elderly counterparts. The current management and aggressive risk factor modification are quite good and the overall mortality is lower in young adults with ACS compared to their elder counterparts. Primary preventive measures aimed at preventing our youth from adopting tobacco use should be implemented nationally. PMID- 18431892 TI - The effect of the public or private status of health care facility in acute coronary syndrome: data from Thai ACS Registry. AB - BACKGROUND: Few data showed the differences between public and private hospitals in management practices and outcomes of patients with acute coronary syndrome. Furthermore, no data is available in Thailand. OBJECTIVE: To determine the patients' characteristics, management practices, and in-hospital outcomes differences between public and private hospitals in Thailand for patients with acute coronary syndrome. MATERIAL AND METHOD: Data from the Thai Acute Coronary Syndrome Registry (TACSR), which was a prospective observational study on ACS in Thailand from 2003 to 2005, was used. This registry provided clinical characteristics, medical management and outcomes of patients with ACS during hospitalization. All data were then compared based on type of admitting hospitals; public and private hospitals. To determine the relationship between type of hospital and major cardiac outcomes, multivariate logistic regression analysis was performed and represented as odd ratio (OR) and 95% confidence interval (95%CI). RESULTS: Eight thousand one hundred sixty four patients were admitted to public hospitals (n = 13), and 1,209 were admitted to private hospitals (n = 4). Patients in public hospitals were older (65.4 +/- 12.1 vs. 63.4 +/- 13.3 years, p < 0.001) and more female gender (41.7% vs. 30.1%, p < 0.001). Diagnosis of acute ST-elevation myocardial infarction were lower in public hospitals compared to private hospitals (39.6% vs. 50%, p < 0.001). After adjusting for baseline patient characteristics and management, in-hospital outcomes were higher in public hospitals for total mortality (13.6% vs. 5.9%, OR 2.3, 95%CI 1.76-3.12, p < 0.001), cardiac mortality (10.6% vs. 4.8%, OR = 2.1, 95%CI 1.55-2.91, p< 0.001) and major bleeding (6.3% vs. 3.2%, OR = 2.1, 95%CI 1.48-3.23, p < 0.001). Compared with the patients in the public hospital, patients in the private hospitals were more likely to undergo coronary angiography, percutaneous coronary intervention and coronary bypass grafting. CONCLUSION: In Thailand, management of patients with acute coronary syndrome is influenced by the public or private status of the hospitals. Patients were more likely to undergo coronary angiography and coronary revascularization procedures in private hospitals. The length of hospital stays and in-hospital mortality was higher in public hospitals. PMID- 18431893 TI - Effect of un-fractionated heparin and low molecular weight heparin on hospital mortality in patients with non ST elevation acute coronary syndrome (ACS). AB - BACKGROUND: The Thai ACS registry is a multi-center prospective registration that describes the epidemiology, management practices and in-hospital outcomes of patients with acute coronary syndromes. OBJECTIVE: Study the registry difference in hospital outcomes about cardiac death and length of stay between low molecular weight heparin (LMWH) and un-fractionated heparin (UFH). MATERIAL AND METHOD: This is an observational descriptive study. The authors collected data from the database of the Thai ACS registry. RESULTS: There were 233 of 3963 cases (5.9%) with cardiac death in the present study. Cardiac death in the non-ST elevated myocardial infraction (NSTEMI) group was larger than in the UA group (7.6% vs. 2.4%, p-value < 0.001). The heparin group had more cardiac death than the LMWH group (9.3% vs. 5.2%, p-value < 0.001). NSTEMI with heparin treatment had more cardiac deaths than LMWH treatment (11.8% vs. 6.8%, odd ratio 1.8). UA with heparin treatment had more cardiac deaths than LMWH treatment (4.0% vs. 2.0%, odd ratio 2.0). NSTEMI had a longer length of stay than UA (56.9% vs. 44.7%, p-value = 0.001). The heparin group had a longer stay than LMWH (58.8% vs. 51.7%, p-value < 0.001). CONCLUSION: Low molecular weight heparin had benefit over un fractionated heparin in reduction of hospital mortality and length of stay in both unstable angina and non-ST elevation myocardial infarction. PMID- 18431894 TI - Educate, communicate, coordinate. PMID- 18431895 TI - Mucositis management. PMID- 18431896 TI - National guidelines for palliative care: A roadmap for oncology nurses. AB - Patients with cancer have significant needs for palliative care, including pain and symptom management and psychosocial and spiritual support. The experience of cancer has an impact on family caregivers as well, and palliative care needs exist from diagnosis through survivorship and end-of-life care. Oncology nurses have opportunities to integrate palliative care into disease-focused care. Clinical practice guidelines developed by the National Consensus Project for Quality Palliative Care and preferred practices defined by the National Quality Forum serve as a framework to guide nurses in this area. These national guidelines can serve as a roadmap to develop clinical services that will serve patients and families. PMID- 18431897 TI - Expectant management as a treatment for early-stage prostate cancer. PMID- 18431898 TI - Osteonecrosis of the jaw. PMID- 18431899 TI - Information for patients. Constipation in the cancer patient. PMID- 18431900 TI - Cardiovascular late effects. AB - Cancer survivors who have received cardiotoxic chemotherapy or mediastinal radiation have an increased risk of developing cardiovascular late effects. Cardiac functioning should be evaluated during follow-up exams based on the patient's risk factors, cardiac symptoms, and type of cancer treatment received. Signs and symptoms indicative of cardiovascular late effects include dyspnea, fatigue, tachycardia, nonproductive cough, neck vein distention, coarse breath sounds, and pedal edema. PMID- 18431901 TI - Epothilones. PMID- 18431902 TI - [Fever without a source. Diagnostic difficulties in invasive bacterial diseases in children observed on bacterial meningitis model]. AB - Acute infection of the central nervous system is the most common cause of fever associated with signs and symptoms of CNS disease in children. Unfortunately most of these symptoms are non-specific. Specific signs appear very often late in the course of illness. 761 cases of bacterial meningitis in children above 1 month of life were investigated. Only 46.1% of cases were correctly diagnosed during the first doctor's exam. We found that correct diagnosis depends on time between the first symptoms of illness and the first physician visit. If this period of time is below 16 hours the risk of wrong diagnosis is very high due to lack of specific signs of CNS infection. In many cases only fever is present then. PMID- 18431903 TI - [Fever of unknown origin--pyelonephritis? Pneumonia? Mastoiditis? Ethmoiditis?]. AB - Fever of unknown origin is alarming phenomenon in childhood. Diagnostic procedures should be focused on severe bacterial infection: pyelonephritis, pneumonia and other respiratory tract infections such as mastoiditis and sinusitis in particular ethmoiditis. The principles of treatment and diagnostics were discussed depending on age and general status of the child. PMID- 18431904 TI - [Fever as alarming symptom of infections in children]. AB - Fever in infants and children frequently causes considerable concern for parents and physicians. It is one of the most common reasons that parents seek medical attention for their children, accounting for 1/3 of pediatric office and emergency department visits. About 14-20 percent of febrile children have fever without an apparent source of infection after history and physical examination. Physician concerns stem largely from the recognition that some of these febrile children are at risk for developing serious bacterial illness and may have early stage of an infection, including bacteremia, urinary tract infection (UTI), occult pneumonia or, rarely, early bacterial meningitis. This article reviews the evaluation and management of children with fever which is the only alarming symptom of infection. A cautious approach should still be taken based on the potential for adverse consequences of unrecognized and untreated serious bacterial infection. PMID- 18431905 TI - [Periodic fever, aphtosus stomatitis, pharyngitis, cervical adenopathy syndrome (PFAPA syndrome) in 4-years old girl--case report]. AB - Characteristic feature of PFAPA syndrome is periodic episodes of fever recurring in 21-28 days in infants and young children. Fever is accompanied by aphtosus stomatitis, pharyngitis and cervical adenopathy. Diagnosis of this syndrome are based on typical clinical manifestations, because there are no characteristic changes in laboratory findings. The reason of this syndrome is unknown. We described a case of 4 years old girl with typical manifestations of this syndrome. We excluded others reasons of periodic fever. PFAPA syndrome passed after 2 years of duration after adenothomy. PMID- 18431906 TI - [Management of fever without source in children]. AB - INTRODUCTION: Infectious diseases are the most common cause of fever in children. Viral and bacterial infections of respiratory, gastrointestinal, central nervous system and urinary tract dominates. In the group of children with chronic fever in some cases inflammatory bowel diseases, immunodeficiency syndromes and infections of rare pathogens are diagnosed. AIM: Authors analysed retrospectively 10 children with chronic fever treated in the II Department of Pediatric Gastroenterology & Nutrition, Medical University of Wroclaw. RESULTS: In those children we diagnosed Crohn's disease (1 case), cat scratch disease (1 case), systemic fungial infection in the child with short bowel syndrome (1 case), severe immunodeficiency syndrome (3 cases), intraperitoneal abscess in a child with Crohn's disease (1 case), severe toxic diarrhoea in the course of treatment with cyclosporin (1 case), Yersinia enterocolitica infection (2 cases). CONCLUSION: In the diagnostics of the prolonged fever in children various conditions and diseases ought to be considered. PMID- 18431907 TI - [Nonconvulsive events in children]. AB - BACKGROUND: Epileptic seizures constitute the most important group of paroxysmal events in the developmental age. There are convulsive and nonconvulsive seizures. Nonconvulsive epileptic seizures must be differentiated from nonepileptic paroxysmal events. THE AIM OF STUDY: This paper recalls the importance of non convulsive epileptic seizures in the developmental age. It should be helpful for improving the diagnosis and therapy of epilepsy and quality of life in childhood. MATERIAL AND METHODS: Many papers across 17 years and own 10 years observations in Department of Pediatric Neurology, Jagiellonian Collegium Medicum in Krakow are presented. RESULTS: These scientific and observation data shows a great importance of nonconvulsive epileptic seizures for epileptic children health and for future of epileptology. Its occured as a short events usually, but also as a epileptic syndromes or non-convulsive status eopilepticus rarely. To establish diagnosis parents, teacher, psychologist, nurse and physician observations are necessary. A home video recordings, professional videoEEG rejestrations, and polysomnography play an important role as well. This paper recalls the importance of EEG analysis for proper differential diagnosis of nonconvulsive epileptic seizures. Adequate management with antiepileptic therapy improves epileptic children's life safety and their quality of life. CONCLUSIONS: A dynamic progress is observed in knowledge of nonconvulsive epileptic seizures in children. An practical example of it is evolution of a Panayiotopoulos syndrome recognazing with interest of the International League Against Epilepsy. PMID- 18431908 TI - [A first convulsive seizure in children--diagnostic procedures and treatment]. AB - The Authors discusses in succession: acute symptomatic seizures, diagnostic program age-related (studies in neonatal convulsions and newborn seizures), general treatment principles as well as a seizure recurrence percentage after a single seizure and management after a first seizure. PMID- 18431909 TI - [The diagnostic and therapeutic management in the first cerebral attack in childhood]. AB - Cerebral attacks are the most frequent neurological problem in childhood. The diagnostic and therapeutic management in the first cerebral attack belongs to subjects often discussed in literature but currently it arouses some controversies concerning especially the notions "cerebral attack", "unprovoked seizures" and also the fact when epilepsy should be diagnosed after the first or the second unprovoked attack of seizures and when to start its treatment. The author presents in detail the principles of the management of cerebral attack and its differentiation in children. In relation to the neurological examinations (EEG, cerebrospinal fluid analysis, neuroimaging examination), the author demonstrates both the indications for their performance as well as limitations due to frequently obtained negative results. Among cerebral attacks, after exclusion of symptomatic attacks and febrile seizures, the first epileptic seizure can be diagnosed in 50% of children. The treatment of epilepsy after the first epileptic seizure decreases effectively the number of seizures within the first three years. However, the percentage of children who achieved 3-5-year remission is in these patients similar to that in the group treated after 2 successive attacks of unprovoked seizures. EEG and neuroimaging examinations (cerebral MR more beneficial than CT) belong to the diagnostic canon in the case of a cerebral attack. Changes in neurological examination and also the changes maintaining after focal attack, the age of children with cerebral attack up to 6 months and changes on EEG and in neuroimaging examinations increase the risk of another attack. The selection of the performed laboratory tests depends on an individual case and should be extended when symptomatic nature of cerebral attack is suspected (i.e. acute or chronic encephalopathies). PMID- 18431910 TI - CNS Lyme disease manifestation in children. AB - BACKGROUND: Neurological symptoms develop in 10-20% of children with borreliosis. AIM OF THE STUDY: It was a presentation of clinical manifestation of neuroborreliosis in children. MATERIAL AND METHODS: Children with neuroborreliosis and other neurological diseases were admitted to the University Hospital during 2005-2006 without any selection. Of these 9 patients, there were seven males and two females, ranging in age between 3-17 years. Neurological diagnostic was performed using ELISA Biomedica kit and western blot bands. A 2-6 week sequential treatment with either i.v. ceftazidime or amoxicillin and oral doxycycline or amoxicillin was provided. Children were monitored regularly during the next 4-24 months. RESULTS: The 9 children with borreliosis constitute 0.53% of the pediatric neurology department's patients. The clinical manifestation of LD were usual and unusual from patient to patient. They included three cases of facial nerve paralysis (with bilateral paralysis in one case). In two cases, they included transverse myelitis and in a single case, hemiparesis, meningitis and acute ataxia. Typically, other patients with early stage borreliosis first manifest focal seizures, raising the suspicion that borreliosis could be responsible for triggering seizures. The antibiotic treatment was successful in 7 patients and only partially effective in 2 children with facial nerve paralysis. CONCLUSIONS: The most common symptom of neuroborreliosis in children is motor dysfunction. Acute ataxia may be a clinical presentation of neuroborreliosis. It is probable that borreliosis_triggers seizures in children with EEG abnormalities. PMID- 18431911 TI - [Rare diagnostically difficult causes of acute abdomen in children]. AB - Abdominal pain is a very nonspecific sign that may be encountered in many diseases in children. The differentiation between acute abdomen and other pathologies may cause problems in many situations. The paper summarizes atypical causes of the acute abdomen which are rare but may be seen in the big, working hospital. PMID- 18431912 TI - [Diagnostic dilemma in abdominal pain in children]. AB - Author presents own experience in diagnosis of abdominal surgical diseases in children. Emphasized aspect estimation of pain as main symptom of the disease. Discuss various types and forms of abdominal pain. Special attention gives to difficulties in diagnosis of appendicitis and intussusceptions. PMID- 18431913 TI - [Chosen diagnostical aspects of abdominal disorders in children]. AB - Paper presents chosen diagnostical problems in abdominal disorders in children. Infant dyschezia was discussed as a functional disorder in infants. In older children symptoms like distention, and abdominal pain which were the results of gas overproduction were the main focus. Small bowel bacterial overgrowth and chronic intestinal pseudo-obstruction was discussed. PMID- 18431914 TI - [Challenge in diagnostics of abdominal complaints in children]. AB - In the present review, abdominal pains were discussed as the most common complaints in children. Abdominal pains were divided into acute and chronic, organic and functional. The most common causes of abdominal pain, alarming symptoms (red flags) in medical history and physical examination and indications to urgent surgical treatment were enumerated. An attention was laid on the difficulty in assessment of intensity and localization of the pain in children. The principal role of physician and caregivers as well as diagnostic challenge in the youngest children was emphasized. Diagnostic criteria of gastrointestinal functional disorders accompanied by abdominal pain were analyzed. Diagnostic process in abdominal pain was presented. PMID- 18431915 TI - [Gastrografin challenge test for the management of subileus in children]. AB - BACKGROUND: The appearance of gastrografin in colon within 6 hours after gastric administration rules out the need of surgery in abdominal subileus. This diagnostic management is not routinely applied in children. We present a one-year experience from the department of pediatric surgery. MATERIAL AND METHODS: Between April 2006 and September 2007 children with symptoms of ileus without clear indications for surgery were subjected to the study. Naso-gastric tube was inserted and 20-100 cc of gastrografin was administered. Abdominal radiograph was taken within 4-6 hours. The presence of contrast in colon allowed for conservative management. All remaining children were subjected to surgery. Newborns and children with intussusception or incarcerated hernia were ruled out of the study. RESULTS: The study was implemented in 8 girls and 7 boys ranking in the age between 1 and 17 (mean 11) years. Thirteen children had postoperative obstruction (8 after appendectomy, 5 after other laparotomy). Two children (with Crohn disease and Schoenlein-Henoch purpura) were not operated before. Four children without appearance of contrast in colon were operated. None of the remaining eleven children required surgical intervention. CONCLUSIONS: This management is safe and effective. It brings forward decision for surgery and shortens observation in children who don't require surgical intervention. PMID- 18431916 TI - [Diagnostic difficulties in pediatric abdominal pain with potential appendicitis]. AB - BACKGROUND: Pediatric abdominal pain is one of the most common symptom of children brought to attention of primary care physicians and pediatric surgeons. In many children clinical symptoms may be uncharacteristic and may lead to diagnostic difficulties. AIM: Clinical analysis of children with right lower quadrant abdominal pain suspected of appendicitis or children with periappendicular mass. MATERIALS AND METHODS: The histories of 14 children aged from 18 months to 17 years treated in Pediatric Clinic were analysed. All children were operated because of right lower quadrant abdominal pain or abdominal mass before admission to the Pediatric Clinic or during hospitalization in Pediatric Clinic. Intraoperatively in all children pathologies other than appendicitis were the cause of symptoms. RESULTS: The most often Crohn's disease were recognized (9 children), in 2 cases with concomitant other pathologies (fecal tumor of appendix in one case and with peritoneal abscess after perforation of intestinal wall). Sporadically the inflammation of the mesenterial lymph nodes caused by Yersinia infection suggested appendicitis. In one boy with ulcerative colitis, during exacerbation of the disease appendicitis complicated by rupture and peritonitis was observed. In 18-month old child with right lower quadrant abdominal mass invagination complicated by perforation of the ileum was recognized. In the case of 14-years old boy 6 months after appendectomy we observed mechanical intestinal obstruction complicated by perforation and peritonitis. Carcinoid of the appendix was the cause of abdominal pain in one child. CONCLUSIONS: 1. Appendicitis is the most frequent surgical etiology of the right lower quadrant pediatric abdominal pain. 2. Despite new diagnostic imagines there are no definite criteria to recognize appendicitis, in most cases physical examination and very carefull evaluation of abdominal pain are the most important. 3. All children with periappendipected of Crohn's disease. 4. All children with equivocal presentations of appendicitis and with normal appendix during operation should undergo further diagnostic evaluation. PMID- 18431917 TI - [Long-term observation of children with gastro-esophageal reflux disease]. AB - INTRODUCTION: Gastroesophageal reflux disease (GERD) is frequent disorder of the gastro-intestinal tract. Although in adults GERD is regarded as a chronic disease, in children it is often considered as a transient disorder. AIM: To assess the natural history of GERD in infants and children. MATERIAL AND METHODS: We enrolled 163 children (44% girls and 56% boys) with GERD diagnosed on the basis of esophageal symptoms and positive result of 24-hour pH-monitoring (fraction time - fT > 4.2%). The mean age at the time of the diagnosis was 4.5 year. After 3.5 years from the diagnosis, the patients or guardians filled in a custom-made questionnaire evaluating GERD symptoms: heartburn, belching, vomiting, regurgitations, feeding problems, dysphagia, epigastric pain. Control pH-monitoring was performed in 71 of the patients. The patients were divided into three age groups: group I--below 1 year of age (38 children), group II--between 1 and 4 years of age (35 children) and group III--above 4 years of age (90 children). RESULTS: GERD symptoms disappeared completely in 45%, 14.3% and 13.4% from group I, II and III, respectively. In 38 (54%) patients the control pH monitoring revealed pathologic acid reflux, including 4 children without any GERD symptoms. CONCLUSIONS: The study shows that in children aged above 1 year GERD symptoms are unlikely to subside completely. Moreover, acid gastroesophageal reflux may still be present, despite the absence of the clinical symptoms. PMID- 18431918 TI - [The low prevalence of Helicobacter pylori gastritis in newly diagnosed inflammatory bowel disease children and adolescent]. AB - Data concerning prevalence rate of Helicobacter pylori gastritis in inflammatory bowel disease (IBD) patients is conflicting. We had studied the prevalence of Hp gastritis in newly diagnosed inflammatory bowel disease children before any pharmacological treatment was introduced. Ninety four consecutive children with inflammatory bowel diseased were enrolled into study, mean age 12.9 +/- 3.75 years, including 50 with Crohn's Disease (CD) and 44 with ulcerative colitis (UC). One hundred and four patients (mean age 13.6 +/- 4.2 year) referred for diagnostic evaluation because of recurrent abdominal pain, matched for age, sex and socioeconomic status served as a control. The results revealed a highly statistically lower prevalence of Hp gastritis in children with IBD as compared with controls (9.6% vs. 38.4%, p < 0.0001). Significantly more often Hp gastritis occurred in CD than UC patients. There was no statistical difference in mean age of IBD onset between Hp gastritis positive and negative groups (14.3 +/- 3.75 vs. 13.6 +/- 4.3 yr) was found. Our results show that in newly diagnosed IBD children, Hp gastritis is not unusual, but the prevalence rate is significantly lower comparing to the control group. The low Hp gastritis rate is not related to medical treatment, since the patients were studied before any was introduced. PMID- 18431919 TI - [Body mass index (BMI) or bioimpedance analysis (BIA) --which method enables a better assessment of fat mass in children?]. AB - A comparison of accuracy between electrical bioimpedance analysis (BIA) and body mass index (BMI) for fat mass evaluation in children was the aim of the study. One-hundred and ninety three healthy children (90 females, 103 males) were enrolled. The mean age of the studied population was 11.8 years (+/- 2.2), mean weight equalled to 42.6 (+/- 12.8) kg and mean height-151.1 (+/- 13.5) cm. BIA measurments were performed by multifrequency (1; 5; 50; 100 kHz) BIA 2000M analyzer using BIANOSTIC electrods. Calculations of free fat mass and fat mass were performed using Nutri 4 Data Input Software (Germany). The LMS method for percentile charts of FFM and FM acording to height has been applied and results for boys and girls were drawn separatelly. Percentile charts of FM and BMI were compared. Different shape of curves of FM content and BMI for boys during puberty was noticed. BIA allows for precise fat mass content evaluation in children and its use should be disseminated. PMID- 18431920 TI - [Somatic disease or Munchausen syndrome by proxy? Mother's or child's disease? Diagnostic controversies]. AB - INTRODUCTIONn: The Munchausen Syndrome by Proxy occurs when parents make a child believe he or she suffers from a disease while in fact this is not true. A medical diagnosis ruling out a possible disease of the child does not calm the parents down and only confirms--in their belief--the doctor's incompetence which in turn makes them seek a second opinion. This may be repeated over and over again and be accompanied by administration of medicines, overprotection, introducing a diet, limiting exercise as well as limiting motoric and mental development. AIMS: Authors present diagnostic difficulties in diagnosing the Munchausen syndrome. CASE DESCRIPTION: A 6-year old girl was hospitalized for the first time at the Developmental Age Psychiatry and Psychotherapy Ward of the John Paul II Pediatric Center in Sosnowiec, Poland (until then she had received psychiatric treatment at the Center on an outpatient basis only). Medical history provided by her mother as well as supplied documents show numerous hospitalizations at Pediatric Wards since the girl was 6 weeks old due to intensified vomiting and frequent diarrheas. Since the age of 4, the girl has been provoking vomiting as well as temporarily refusing meals and liquids. COMMENTARY: Authors present diagnostic difficulties in diagnosing the Munchausen syndrome mainly caused by discharging the patient from the Ward on her mother's request as well as non-specific nature of the symptoms. PMID- 18431921 TI - [Syncope in children and adolescents]. AB - The report presents a definition and causes of syncope in children. Syncope differs from other states with loss of consciousness by causes leading to decreased perfusion and resultant transient cerebral dysfunction with decreased muscle tone. The most common causes of syncope noted in almost 15% of children are neurocardiogenic. This group includes vasovagal, carotid sinus reflexive, situational (coughing, dysphagia, micturation and defecation disturbances) and post-exercise syncope. Another group is represented by orthostatic syncope that may be triggered by primary and secondary dis-autonomy, decreased blood volume (hemorrhage, diarrhea, Addison's disease), some medications and substances of abuse (alcohol). An important group, accounting for 2%-6% of all cases, are cardiogenic syncope, caused mainly by congenital/acquired obstructive cardiac sub and valvar heart defects, various cardiomyopathies, some heart tumors (e.g. myxoma), exudative pericarditis, pulmonary embolus and hypertension, congenital and acquired coronary anomalies, various significant brady-tachyarrhythmias (sick sinus syndrome, supra- and ventricular tachycardias, congenital and acquired atrio-ventricular blocks). Subclavian steal syndrome as the cause of syncope is exceptional in children. Syncope does not include loss of consciousness due to neurological and metabolic (hypoglycemia) causes, hypoxia, hyperventilation with hypocapnia or CO intoxication. Differential diagnosis should also include pseudo syncope (hysteria). Preliminary diagnostic management should include a detailed medical history, including family history, on the frequency and circumstances of syncope, sudden deaths, a physical exam with orthostatic assessment of peripheral blood pressure and standard ECG (heart rate, intraventricular and atrioventricular conduction defects, cardiac hypertrophy, arrhythmias, L-QT, changes in ST-T). Further specialist tests depend on preliminary findings. PMID- 18431922 TI - [Syncope in children]. AB - Syncope is a very common problem during childhood presenting to emergency departments. It generates a great deal of anxiety among children and their parents because of the fear that syncope are at risk for sudden death. Neurocardiogenic (vaso-vagal) syncope are the most common, while cardiac type are rare. The vaso-vagal syncope are secondary to irregularities in an autonomic reflex resulting in an abnormal relaxation of the blood vessels. This is non-life threatening form of syncope in differ cardiac type that can be very dangerous. In cardiac syncope causes can be secondary to obstruction to blood flow (Tetralogy of Fallot, cardiomyopathies, aortic stenosis), heart rhythm abnormalities (WPW, L QTS, A-V block) as well as ischemic heart disease secondary to an anomalous coronary artery. The cardiac syncope are rare (6%) but the most inconvenient because they can be life-threatening. PMID- 18431923 TI - [Adaptation of cerebral blood flow to orthostatic stress in children with neurally mediated syncope]. AB - In the pathogenesis of neurally mediated syncope the key role play autonomic dysregulation of cardiovascular function. The clinical picture of syncope alone may not be sufficient for precise diagnosis, therefore the assessment of hemodynamic changes and cerebral function is necessary. The interdisciplinary management resulted in development of functional methods of examination of cardiovascular responses with special attention to head-up tilt test. The presentation of simultaneous cerebral blood flow monitoring during orthostatic stress resulted in the progress in evaluation of syncope and explanation of pathomechanism. The changes in the velocities in the middle cerebral artery before syncope were detected with Doppler examination. Results of several examinations are however not consistent. The development of new methods of examination of cerebral blood flow as autonomic nervous system as well, since autonomic balance disturbances were detected in children with neurally mediated syncope. PMID- 18431924 TI - [Cardiac syncope in children]. AB - INTRODUCTION: Syncope are serious problems in paediatrics. Approximately 15% of children and adolescents at the age of 8-18 years experience at least one episode of syncope. Neurocardiogenic syncope are the most common (60-80%). Cardiogenic syncope are rare (6%), although they require special attention due to their background. AIM OF THE STUDY: Evaluation of incidence, causes, methods of treatment and prognosis in cardiogenic syncope in children and adolescents. MATERIAL AND METHODS: Retrospective analysis was made in 18 patients with syncope and different pathology of cardiovascular system. Patients were chosen among 766 children treated in the University Hospital for Children in Cracow from January 2005 to June 2006 due to syncope or pre-syncope. The patients were divided into 2 groups depending on the main cause of syncope: Gr I n = 6 due to arrhythmias without structural heart disorders and Gr II n = 12 due to heart diseases or defects. In all patients physical exam and additional test were analysied carefully (ECG, Holter study, chest X ray, ECHO, angio-TK in 1 patient, and catheterisation with angiography in 3 children). RESULTS: Cardiogenic syncope appeared more frequently in children with structural abnormalities in the heart (Gr II--12 pts/ 66%), rather than in children with arrhythmias (Gr I--6 pts / 33%). Family history was positive in both groups in 17%. Median age at the moment of first episode was 10.3 y +/- 5.0 in Gr I and 12.8 y +/- 3.0 in Gr II. Syncope in Gr II were frequently (33%) initiated by an effort or stress. In Gr I physical examination did not reveal serious symptoms except for irregularity of heart rate, and general condition of children was good (NYHA I). In 5 pts (58%) in Gr II heart murmur and prominent impulse of the left ventricle were found. Symptoms of congestive heart failure were observed in 4 pts (33%) and their condition was poor (III/IV class according to NYHA). In ECG and/or Holter study in Gr I arrhythmias and/or long QTc were found. Pharmacotherapy was sufficient in treatment of serious arrhythmias and prophylaxy of syncope. In Gr II in most cases hypertrorepolarization abnormalities were found. In these children treatment was multidirectional and included both antiarrhythmic drugs, implantation of cardioverter defibrillator (ICD), operation and even heart transplantation. CONCLUSIONS: Cardiogenic syncope are rare causes of loss of consciousness and indicate on serious pathology in the cardiovascular system. Heart structural diseases or defects influence severity on clinical course, complex treatment and serious prognosis while in children with arrhythmias pharmacotherapy is effective and prognosis is usually good. PMID- 18431925 TI - [Shall we recognize chronic renal insufficiency as a pediatric controversy?]. AB - Introduction in 2002 the new, 5 - degree classification of chronic renal disease which has been based upon calculation of glomerularfiltration rate (eGFR)--on the one hand took note of the problem of kidney injury and decrease of active nephrons' number which may accompany various renal diseases--on the other- allowed to define the risk factors, which include first of all--hypertension and persistent proteinuria. Chronic renal disease is diagnosed in each clinical case, where a decrease of glomerular filtration rate below 90 ml/min/1.73m2 had occurred with or without kidney injury or when a decrease of glomerular filtration rate maintains for at least 3 months on the level < 60 ml/min/1.73 m2. Delayed diagnosis of chronic kidney disease leads to manifestation of chronic renal failure symptoms and excludes an effective nephroprotective treatment. In the face of a large number of potential causes of chronic renal disease which may be encountered by a pediatrician, all children which are numbered among the high risk group--should have eGFR calculated--initially according to the simplest Schwartz formula. Setting of a diagnosis of chronic renal failure only on the basis of serum creatinine concentration doesn't allow to notice hyper-filtration phenomenon and should not be a daily clinical practice. Fundamental approach of therapeutical management in in children with chronic renal disease is slowing down the disease progression and/or elimination or modification of some risk factors. Each child with diagnosed chronic renal disease should be referred to specialist outpatient pediatric nephrology clinic. PMID- 18431926 TI - [Chronic kidney disease--diagnostic problems]. AB - Current data indicate that the diagnosis of chronic kidney disease (CKD) is often late. It is estimated that 20-50% of adult patients start dialysis therapy without prior contact with a nephrologist. In contrast to early referral patients (both children and adults), late referral to nephrologist is related to poorer clinical status, hypertension and more severe uremic symptoms. Late referral is also associated with longer hospitalization, higher treatment costs and early deaths on dialysis. Factors influenced on the late diagnosis of CKD in children, as well as the late referral to a nephrologist, are discussed in the present review. PMID- 18431927 TI - [Systemic consequences of chronic kidney disease in children]. AB - Major consequence of chronic kidney disease is multiorgan dysfunction. Severity of clinical symptoms is in general associated with degree of renal dysfunction, however is not uniformly distributed among various organs and systems. Tailored conservative treatment is aimed to prophylaxis and slowing the progress of organ dysfunction. Nevertheless, the main target is to maintain the best available quality of life until renal replacement therapy is necessary. PMID- 18431928 TI - [Controversy in chronic primary glomerulonephritis treatment in children]. AB - Medical and ethical problems connected with treatment of chronic primary glomerulopathy in children were discussed. This treatment is difficult and medical controversial. Two cases of patients with ineffective treatment and severe side effects were described. Psychological problems that may arise from chronic serious disease were emphasized. The question of co-operation doctor with adolescent patient was analyzed. PMID- 18431929 TI - [Next episode of wheezing in infant--what to do? Natural history of wheeze]. AB - Wheezing illness is a common problem during first year of life and is observed in the one third of this age group. During infancy, in most cases wheeze is episodic and transient. In some of them, wheeze is chronic and persistent. Retrospective clinical studies show that asthma is diagnosed in later life in the part of persistent wheez-ers. Conducted in the past decade prospective epidemiological studies revealed significant factors related to the development of asthma from the group of persistent wheezers in early period of life. It can be believed that childrens early identification with increased risk of asthma development allows to introduce effective preventive programs. PMID- 18431930 TI - [Next episode of wheezing in infant--what to do?]. AB - The choice of diagnostic and therapeutic options in infant with recurrent wheezing remains difficult. Number of episodes, severity of respiratory symptoms and co-morbidities should be analyzed. Viral infections are the risk factors of recurrent wheezing. Atopy predispose to asthma development. Most studies focus on assessment of efficacy of inhaled glucocorticoids, antileukotriens and cromones. Inhaled steroids are considered as the most effective. The initiation of therapy is recommended in children with at least 3-4 episodes or after one, but severe exacerbation. The treatment with inhaled steroids decrease symptoms, but has no influence on natural course of disease. PMID- 18431931 TI - [Next wheezy episode in infant--what to do?]. AB - There were presented several aspects of epidemiology, diagnosis and therapy in infants with recurrent bronchial obturation (wheezing). It was emphasized the crucial role of medical history and some laboratory tests. Pharmacological and non-pharmacological methods of early intervention in wheezing neonates were also discussed. PMID- 18431932 TI - [Retrospective analysis of coexistence of acidic gastroesophageal reflux in children with atopic bronchial asthma]. AB - INTRODUCTION: Epidemiologic studies consistently demonstrate strong associations between gastro-esophageal reflux disease (GERD) and bronchial asthma also in children. A significant percentage of patients with GERD may experience extra esophageal manifestations like cough or bronchspasm, which are typical symptoms of uncontrolled asthma. OBJECTIVE: retrospective analysis of prevalence and severity of GERD in children with atopic bronchial asthma and influence of proton pomp inhibitor (PPI) therapy in management of asthma. METHODS: The retrospective analysis of children with uncontrolled bronchial asthma between 6 and 18 years of age was performed. Thirty (21 boys) children were evaluated for acidic gastroesophageal reflux using 24-h esophageal pH-monitoring. Age, area of residence, presence and clinical severity of GERD, nocturnal episodes of GERD, pulmonary function parameters were analyzed. The effect of PPI therapy on modification of asthma treatment and lung function was assessed. RESULTS: Acidic gastroesophageal reflux was diagnosed in 17 (56%) children (fraction time of pH < 4 above 4.2%). Most of those patients (88%) came from urban area. Mean age at diagnosis of GERD was 10.5 years of age. Mild GERD was diagnosed in 13 children, moderate--in 4 children. Nocturnal episodes of reflux were present in 9 children (53%). Differences between lung function parameters (FEV1, FVC, PEF) before and after PPI treatment were not statistically significant, although their mean values were about 10% higher after treatment. PPI therapy allowed reduction of inhaled steroids dose in 24% of children; LABA were withdrawn in 3 children. CONCLUSION: The association of GERD with atopic asthma in children is common. It seems to be reasonable to perform work up of pathologic GER in children with uncontrolled asthma. PMID- 18431933 TI - [Wheezing in very low birth weight infants: sequence of early neonatal lung injury or increased susceptibility for allergic reactions? Follow-up study up to age of 5-7 years]. AB - BACKGROUND: Prevalence of wheezing during infancy and pre-school age among very low birth weight (VLBW) infants is high. There is too little data to determine exactly the cause of wheezing in this group of children--it may result from early neonatal lung injury or increased susceptibility for allergic reactions. AIM: Analysis of wheezing episodes up to 5-7 years of age in a group of VLBW infants and recognition of risk factors. METHODS: A sample of 104 children aged 5-7 years with birth weight < or = 1500g was prospectively evaluated. The standardized ISAAC (International Study of Asthma and Allergies in Childhood) protocol was used to evaluate the number and quality of wheezing episodes. Total IgE level, specific IgE, lymphocyte Th1/Th2 ratio and skin prick tests (SPT) were performed. Children were divided into 3 groups: without wheezing, with episodes of wheezing in the history (more than 12 months previously), and with persistent episodes of wheezing. RESULTS: Episodes of wheezing were diagnosed in 52 (50%) cases: 27% of children showed signs of wheezing during 12 previous months and 23% of children reported wheezing only in their past history. Children with persistent wheezing presented more frequently than other groups: positive family history for atopy, allergic rhinitis, and positive allergic laboratory markers. Children with wheezing only during early infancy had bronchopulmonary dysplasia and more severe respiratory distress syndrome after birth. CONCLUSION: Prevalence of wheezing during infancy and preschool age among VLBW infants is high. Chronic and recurrent episodes of wheezing are more directly related to atopy than derive from neonatal problems. PMID- 18431935 TI - Hiring the right person. PMID- 18431934 TI - Where art thou content? PMID- 18431936 TI - Is it a screening or diagnostic mammogram? PMID- 18431937 TI - The impact leadership has on training and change. AB - * Training alone often misses the mark. One tenth of one percent of all change initiatives lead to any kind of positive sustainable change that can be attributed to the effect of the training. One out of every thousand training programs, culture change initiatives, experiential interventions, actually in-and of-themselves appear to have the intended effect. * The results an organization is experiencing (good or bad) are a product of the organizational/work design and management system. Results do not create themselves. They are a consequence, a determined outcome of all the ingredients of the organization. * Many leaders deal with the symptoms of a problem instead of focusing on the root cause. * Working on oneself and then leading the training (whatever it is) is the best way to ensure results for you and your team. PMID- 18431938 TI - Personality assessments as a workforce diversity tool. AB - * Employee recruitment, retention, and workforce diversity are essential ingredients in a successful radiology department. Since potential new hires and existing employees do not come with labels which accurately describe their innate characteristics, radiology managers need to use objective tools such as personality assessments in order to properly evaluate prospective employment candidates. * These tools have traditionally been under utilized in the healthcare industry, but when used as part of a complete employee selection program they have the potential of ensuring a cohesive radiology department filled with diverse and skilled professionals. * Legal concerns and time constraints withstanding, use of personality assessments as a workforce diversity tool will aid in building an integrated department culture which values the varying skills and attributes of every employee. The result is a radiology department with high levels of employee retention and satisfaction. PMID- 18431939 TI - The radiology report: a new look at an old standard. PMID- 18431940 TI - Critical perspectives on cultural competence as a strategic opportunity for achieving high performance in healthcare organizations. AB - * Cultural proficiency is a critical component of diversity competence. Diversity competency attends to issues of cultural proficiency and links cultural competence to issues of diverse representation and organizational inclusivity. * Developing and applying cultural competencies in diagnostic and invasive imaging services provides strategic opportunities to experience better patient cooperation, increase patient satisfaction, reduce medical errors, reduce patient complaints, and improve service recovery. * New Jersey recently passed legislation that requires 16 hours of cultural competency education as a condition of licensure to practice medicine. Four other states have similar legislation pending. The state of Washington passed legislation requiring all state accredited programs to include cultural competency education. PMID- 18431941 TI - Searching for savings. PMID- 18431942 TI - The economic and clinical benefits of portable head/neck CT imaging in the intensive care unit. AB - * There is a 13% morbidity associated with transporting critically-ill patients outside of the ICU. The incidence of adverse events during transport specifically for CT imaging is as high as 71%. The objective of this study was to assess the feasibility and cost-effectiveness of a portable CT scanner designated to perform bedside imaging in the ICU. * A fully mobile 8-slice head/neck CT scanner was evaluated for efficiency and personnel allocation. The return-on-investment for the purchase of the portable scanner was calculated. * Data demonstrates that the introduction of a portable CT scanner in the ICU is feasible and cost-effective. At the Cleveland Clinic in Mayfield Heights, Ohio, the portable scanner provided a full return-on-investment within the first 6.9 months of its operation, an internal rate of return of 169%, and a 5 year expected economic benefit of $2,619,290. PMID- 18431943 TI - Are your vendors competent? PMID- 18431944 TI - Treasure hunt. PMID- 18431945 TI - Influence of a horizontal approach on the mechanical output during drop jumps. AB - This study investigated the influence of a horizontal approach to mechanical output during drop jumps. Participants performed drop jumps from heights of 15, 30, 45, and 60 cm with zero, one, two, and three approach steps. The peak summed power during the push-off phase changed quadratically across heights (6.2 +/- 0.3, 6.7 +/- 0.4, 6.4 +/- 0.4, and 6.0 +/- 0.4 kW, respectively) driven by the ankle joint response. Summed peak power was 10% greater with an approach attributed to the knee joint response. Downward phase dorsiflexion (31%), knee flexion (35%), and peak vertical force (32%) increased with drop heights. Vertical approach force (22%) increased, while knee flexion (11%) and downward duration (17%) decreased. An approach may improve drop jump training for explosive tasks. PMID- 18431946 TI - Patterns of physical activity and overweight among 7-13-year-old Russian children: a 7-year nationally representative monitoring study. AB - This is a repeated cross-sectional study of overweight and physical activity (PA) and inactivity patterns of successive samples of 7-13-year-old Russian children who were surveyed six times between 1995 and 2002 as part of the Russian Longitudinal Monitoring Survey. This analysis focused on moderate/vigorous (MOD/VIG) PA (e.g., active commuting to school, during school MOD/VIG PA as part of physical education classes, and/or before/after school MOD/VIG PA) and inactivity patterns (e.g, television viewing habits) obtained through parent proxy-reports of 3,507 boys (M age = 10.1 years, SD = 2.0; body mass index = 17.5 kg/m2, SD = 3.0) and 3,273 girls (M age = 10.1 years, SD = 2.0; body mass index = 17.2 kg/m2, SD = 3.0). Time spent in MOD/VIG PA appears to be on the rise, while the proportion of those actively commuting to school is decreasing, especially among girls. PMID- 18431947 TI - But I like PE: factors associated with enjoyment of physical education class in middle school girls. AB - The current study examined associations between physical education (PE) class enjoyment and sociodemographic, personal, and perceived school environment factors among early adolescent girls. Participants included 1,511 sixth-grade girls who completed baseline assessments for the Trial of Activity in Adolescent Girls, with 50% indicating they enjoyed PE class a lot. Variables positively associated with PE class enjoyment included physical activity level, perceived benefits of physical activity, self-efficacy for leisure time physical activity, and perceived school climate for girls' physical activity as influenced by teachers, while body mass index was inversely associated with PE class enjoyment. After adjusting for all variables in the model, PE class enjoyment was significantly greater in Blacks than in Whites. In model testing, with mutual adjustment for all variables, self-efficacy was the strongest correlate of PE class enjoyment, followed by perceived benefits, race/ethnicity, and teachers' support for girls' physical activity, as compared to boys, at school. The overall model explained 11% of the variance in PE class enjoyment. Findings suggest that efforts to enhance girls' self-efficacy and perceived benefits and to provide a supportive PE class environment that promotes gender equality can potentially increase PE class enjoyment among young girls. PMID- 18431948 TI - Walking for health in pregnancy: assessment by indirect calorimetry and accelerometry. AB - The purpose of this study was to examine RT3 accelerometer activity counts and activity energy expenditure of 36 pregnant women at 20 and 32 weeks' gestation during treadmill walking and free-living conditions. During treadmill walking, oxygen consumption was collected, and activity energy expenditure was estimated for a 30-min walk at a self-selected walking pace. The number of min it would take a pregnant woman to meet exercise recommendations (i.e., kcal/week) were calculated. Preliminary activity count cut points at a self-selected walking pace were then estimated and applied in interpreting free-living data. For the treadmill walking condition, the self-selected walking pace significantly decreased from 20 to 32 weeks' gestation. Additionally, few women (< 12% each day) met physical activity guidelines in the free-living condition. Encouraging pregnant women to walk for 30-40 min per day at a self-selected walking pace may be an appropriate public health recommendation. PMID- 18431949 TI - The end-state comfort effect in young children. AB - The end-state comfort effect has been observed in recent studies of grip selection in adults. The present study investigated whether young children also exhibit sensitivity to end-state comfort. The task was to pick up an overturned cup from a table, turn the cup right side up, and pour water into it. Two age groups (N = 20 per group) were studied: preschool children (2-3 years old), and kindergarten students (5-6 years old). Each child performed three videotaped trials of the task. Only 11 of the 40 children exhibited the end-state comfort effect, and there were no differences between age groups. Results revealed the emergence of five different performance patterns, none of which were consistent with sensitivity to end-state comfort. The findings have implications for the advance planning of manual control in young children. PMID- 18431950 TI - Construct validation of physical activity surveys in culturally diverse older adults: a comparison of four commonly used questionnaires. AB - The purpose of this study was to establish validity evidence of four physical activity (PA) questionnaires in culturally diverse older adults by comparing self report PA with performance-based physical function. Participants were 54 older adults who completed the Continuous Scale Physical Functional Performance 10-item Test (CS-PFP10), Physical Activity Scale for the Elderly (PASE), HAMPS Physical Activity Questionnaire for Older Adults, Yale Physical Activity Survey (YPAS), and modified Baecke questionnaire. The total PASE score, three outcome scores for the CHAMPS, and three summary indices for the YPAS were significantly correlated with total CS-PFP10 score. The modified Baecke exhibited no correlations with CS PFP10 scores. The PASE, CHAMPS, and YPAS appear to be the most valid PA self report questionnaires for culturally diverse older adults. PMID- 18431951 TI - Effects of dispositional ability conceptions, manipulated learning environments, and intrinsic motivation on persistence and performance: an interaction approach. AB - The present study used an interaction approach to investigate how individuals' dispositions about ability as incremental or fixed (entity), manipulated learning environments, and intrinsic motivation affect persistence and performance on a challenging, novel motor skill. Seventy-two female college students who were assigned to either an incremental or entity learning condition for instruction and practice completed questionnaires and attempted to learn the skill. The results indicated that participants oriented toward incremental beliefs were likely to be more intrinsically motivated. Participants who were intrinsically motivated were likely to perform better on a final skill test. The significant interaction effect on persistence suggests that participants who were highly intrinsically motivated in the incremental condition persisted longer than those in the entity condition. PMID- 18431952 TI - An examination of the multidimensionality of situational interest in elementary school physical education. AB - It has been demonstrated that situational interest in physical activity may derive from five dimensional sources, Novelty, Optimal Challenge, Attention Demand, Exploration Intent, and Instant Enjoyment. The purpose of this study was to examine the multidimensional sources in elementary school physical education. The five dimensions were measured in 5,717 students in third, fourth, and fifth grades from a random sample of 30 elementary schools. Students' responses were randomly divided into two samples for a two-step confirmatory factor analysis. The results confirmed that the five dimensions are primary sources of situational interest for elementary school physical education. The findings implied that situational interest should be taken into account as a necessary curricular component in elementary physical education. PMID- 18431953 TI - Examining curricular coherence in an exemplary elementary school program. AB - A coherent curriculum is characterized by visible connections between purposes and experiences so that students acknowledge the content's immediate value. This study examined an exemplary elementary physical education curriculum for coherence components. Research questions examined the role of coherence in connecting and engaging students meaningfully in physical education. Observations and interviews were conducted to collect qualitative data in one program for 22 weeks. Data were analyzed using open, axial, and selective coding. Results described two units, Balls Skills, leading to modified basketball, and Scooter City, a theme-based unit emphasizing student choice and responsibility. Students reported that both units were enjoyable. Although the Balls Skills unit was well planned, taught, and managed, some students commented that the skill and games content was valuable only in basketball. In the Scooter City unit, students identified numerous connections to out-of-school activities that enhanced content value. Comparisons with Beane's coherence criteria suggested that students valued Scooter City based on concrete connections to their lived experiences. PMID- 18431954 TI - Heart rates of high school physical education students during team sports, individual sports, and fitness activities. AB - This study examined how activity type influenced heart rates and time spent in target heart rate zones of high school students participating in physical education classes. Significantly higher average heart rates existed for fitness (142 +/- 24 beats per minute [bpm]) compared to team (118 +/- 24 bpm) or individual (114 +/- 18) activities. Similar results occurred for the percentage of activity time spent within a target heart rate zone (fitness 81.7 +/- 15.9%, individual 68.4 +/- 30.5%, and team 60.6 +/- 30.5%). Boys attained higher heart rates during team activities, while female students had higher rates during individual activities indicating male and female adolescents respond differently to activity types. The highest mean heart rates were observed during fitness activities. PMID- 18431955 TI - Self-efficacy and outcome expectancy in beginning weight training class: their relations to students' behavioral intention and actual behavior. AB - This study was an initial attempt to investigate the relationships among self efficacy, outcome expectancy, behavioral intention, and actual behavior over time in a beginning weight training class. A total of 109 participants completed questionnaires assessing their self-efficacy, outcome expectancy, and intentions for future weight training. Their attendance and workout logs were calculated as a measure of actual behavior. Correlation analyses revealed significant positive relationships among all the variables during both program periods. Multiple regression analyses indicated that outcome expectancy initially played a more important role than self-efficacy in predicting behavioral intention and actual behavior. However, self-efficacy appeared more influential at midprogram. Findings of this study can enhance our understanding of the determining factors for individuals' motivated behaviors in college physical activity classes. PMID- 18431956 TI - Italian women's television coverage and audience during the 2004 Athens Olympic Games. AB - This study aimed at determining the amount of Italian television coverage dedicated to men's and women's sport and the number of male and female viewers during the 2004 Summer Olympic Games. AUDITEL-AGB Nielsen Media Research Italia provided the TV airtime data for the sport events broadcast, which were classified into three categories: men-only, women-only, and mixed-gender. The viewer sample was divided by age and gender and included three audience parameters: mean audience, share, and appreciation index. The last item is calculated from the program duration, audience permanence, and the share related to programs broadcast by other Italian channels. In particular, the appreciation index was used to investigate the relationship between (a) the viewer's and sport participants' gender and (b) the effect of Italian participation and expectations for sport achievement. The data showed that Italian male athletes outnumbered their female counterparts. Women's sport was allotted significantly less airtime than men's sport, but this imbalance was not proportional to the gender difference in the overall athletes' participation in the Games. Although the female audience represented 45% of the total, a significantly higher male mean audience and share were found. On the other hand, the appreciation of men's and women's sport events was balanced after equating for differences in media coverage. Moreover sport achievement might have a positive effect on the trend toward equality of gender coverage, indicating that a global multisport event such as the Olympic Games is central to communicating a national image and identity independently of the athletes' gender. PMID- 18431957 TI - Task switching effects in anticipation timing. PMID- 18431958 TI - Self-controlled feedback in 10-year-old children: higher feedback frequencies enhance learning. PMID- 18431961 TI - Probiotics for the prevention of necrotizing enterocolitis. AB - Necrotizing enterocolitis (NEC) remains a significant cause of morbidity and mortality for low birth weight premature infants. Prematurity, ischemia, formula feeding, and bacterial colonization are risk factors for the self-perpetuating cycle of damaged intestinal epithelia, inflammation, bacterial entry, sepsis, and shock that characterizes NEC. Probiotics are food supplements containing live bacteria that benefit the recipient by improving the microflora balance within the intestine. Several studies suggest that the administration of probiotics may have a prophylactic effect for NEC and may reduce morbidity and mortality rates for low birth weight infants. PMID- 18431959 TI - Allergy diagnostic testing: an updated practice parameter. PMID- 18431962 TI - Meconium aspiration syndrome. AB - Meconium aspiration syndrome (MAS) is one of the most common conditions associated with aspiration during the newborn period. MAS can be defined as respiratory distress in a neonate born through meconium-stained amniotic fluid (MSAE) with symptoms that cannot otherwise be explained. It can be characterized by early onset of respiratory distress in term and near-term infants delivered through MSAE. Early presentation includes respiratory symptoms such as respiratory distress, poor lung compliance, hypoxemia, and radiographic findings of hyperinflation and patchy opacifications. This article discusses the pathophysiologv of MAS, historical and current prevention strategies, current management strategies, and prognoses for infants born through MSAE. PMID- 18431963 TI - The changing role of parents in neonatal care: a historical review. AB - Over the past century, improvements in technology and neonatal care techniques have dramatically reduced infant mortality rates. While this progress continues, a growing body of literature supports the significant role that parents play in the development of infants, particularly within the hospital setting. Throughout much of the twentieth century, various barriers prevented many parents from participating in thc care of their neonates, negatively influencing infant outcomes. Today parental involvement in neonatal carc has become a key part of a larger family-centered care model. This historical review describes how past neonatal care practices affected the roles of parents, from absence or indirect involvemcnt to the important involved roles of today. Understanding current trends in relation to these past experiences may encourage the formulation of family-centered care practices now and in the friture. PMID- 18431964 TI - Conscientious objection: a potential neonatal nursing response to care orders that cause suffering at the end of life? Study of a concept. AB - This article is an exploratory effort meant to solicit and provoke dialog. Conscientious objection is proposed as a potential response to the moral distress experienced by neonatal nurses. The most commonly reported cause of distress for all nurses is following orders to support patients at the end of their lives with advanced technology when palliative or comfort care would be more humane. Nurses report that they feel they are harming patients or causing suffering when they could be comforting instead. We examined the literature on moral distress, futility, and the concept of conscientious objection from the perspective of the nurse's potential response to performing advanced technologic interventions for the dying patient. We created a small pilot study to engage in clinical verification of the use of our concept of conscientious objection. Data from 66 neonatal intensive care and pediatric intensive care unit nurses who responded in a one-month period are reported here. Interest in conscientious objection to care that causes harm or suffering was very high. This article reports the analysis of conscientious objection use in neonatal care. PMID- 18431965 TI - C-reactive protein in neonatal sepsis. PMID- 18431966 TI - Insulin infusion for the treatment of hyperglycemia in low birth weight infants: examining the evidence. PMID- 18431967 TI - Mission (IM) possible? Nurse civility in the NICU. AB - Nurses will work in a unit and choose to stay because they feel pride in their work and are respected. Eliminating horizontal hostility at the unit level begins with a commitment from the NICU's leadership. Establishing and maintaining an environment of civility is essential if the unit is to successfully recruit and maintain the best staff to care for the patients entrusted to it. PMID- 18431969 TI - I am a [male] nurse. PMID- 18431970 TI - Nursing has changed dramatically just in the last three decades! Interview by Emily Little. PMID- 18431972 TI - Reaching the top: nursing and the Seven Summits. PMID- 18431971 TI - Changing lives through political action as a nursing student. PMID- 18431973 TI - Observe the missing link: pharmacology is safer when you think! PMID- 18431974 TI - Remembering Mrs. Chase. Before there were Smart Hospitals and Sim-Men, there was "Mrs. Chase". PMID- 18431975 TI - Nursing education reinvented. AB - The students at UT Arlington School of Nursing's Smart Hospital' are having a different experience than did the students who worked with "Mrs. Chase" (see page 52). But the end result is the same--to better patient advocacy and outcomes. PMID- 18431976 TI - Industry-sponsored medical education--in the quest for professional integrity and legal certainty. AB - Industry-sponsored medical education is a much disputed issue. So far, there has been no regulatory framework which provides clear and definite rules as to whether and under what circumstances the sponsorship of medical education is acceptable. State regulation does not exist, or confines itself to a very general principle. Professional regulation, even though applied frequently, is rather vague and indefinite, raising the general question as to whether self-regulation is the right approach at all. Certainly, self-regulation by industry cannot and should not replace other regulatory approaches. Ultimately, advertising law in general and the European Directive 2001/83/EC specifically, might be a good starting point in providing legal certainty and ensuring the independence of medical education. Swiss advertising law illustrates how the principles of the European Directive could be implemented clearly and unambiguously. PMID- 18431977 TI - The influence of the rulings of the Constitutional Court on the development of health law in Lithuania. AB - This article highlights some recent developments in the constitutional doctrine of the right to health care in Lithuania, and more in particular the impact of the decisions of the Constitutional Court of Lithuania on the development of health law. The right to health care, enshrined in the Constitution, is both an obligation of the state and an individual right. The Constitutional Court has developed a doctrine of the right to health care, as well a doctrine of certain other constitutional social rights, which is based on the understanding of the close interrelation between the different constitutional rights, the principle of indivisibility and equal importance of these rights, and the presumption of justiciability of social rights. The analysis is based on the jurisprudence of the Constitutional Court. Two cases on the disputes of the legal regulation concerning the pharmaceutical activities are presented in more detail. PMID- 18431978 TI - Constitutional review of the Slovenian health law. PMID- 18431979 TI - Elderly patients as a vulnerable category of the population requiring special legal protection with respect to the provision of medical care. PMID- 18431980 TI - Medical liability and liability of health institutions in Turkish law. PMID- 18431981 TI - Evaluation of the Dutch legislation on euthanasia and assisted suicide. PMID- 18431982 TI - European Court of Human Rights. ECHR 2007/11. Case of Ciorap v. Moldova, 19 June 2007, no. 12066/02 (Fourth Section). PMID- 18431983 TI - European Court of Human Rights. ECHR 2007/12. Case of Kucheruk v. Ukraine, 6 September 2007, no. 2570/04 (Fifth Section). PMID- 18431984 TI - European Court of Human Rights. ECHR 2007/13. Decision as to the admissibility of Yildirim v. Turkey, 11 September 2007, no. 25327/02 (Second Section). PMID- 18431985 TI - European Court of Human Rights. ECHR 2007/14. Case of L. v. Lithuania, 11 September 2007, no. 27527/03 (Second Section). PMID- 18431986 TI - European Court of Human Rights. ECHR 2007/15. Decision as to the admissibility of Eglert v. Sweden, 20 September 2007, no. 39432/04 (Third Section). PMID- 18431987 TI - European Court of Human Rights. ECHR 2007/16. Case of Yakovenko v. Ukraine, 25 October 2007, no. 15825/06 (Fifth Section). PMID- 18431988 TI - European Court of Human Rights. ECHR 2007/17 case of Efendiyeva v. Azerbaijan, 25 October 2007, no. 31556/03 (First Section). PMID- 18431989 TI - Diagnostic criteria for house dust mites sensitized allergic patients. AB - House dust mites (HDM) contain a large number of components that react with IgE in individuals with allergies and induced sensi-tization and allergic diseases. The work aimed to correlate between clinical manifestations of HDM allergic patients and skin prick test (SPT) grading, and to clarify some markers denoting allergic disorders caused by HDM exposure. Thirty allergic patients with positive SPT reaction to HDM were selected, grading of disease severity according to SPT and clinical presentations. The results revealed bronchial asthma (40%), atopic dermatitis (26.7%), rhinitis (20%) and conjuncttivitis (13.3%). Serum ELISA-IgE showed mean level of 398.41 +/- 84.73, 369.87 +/- 86.76, 332.16 +/- 65.57 and 300.25 +/- 79.39 IU/ml respecttively, with highly statistically significant difference between cases and controls (p < 0.001). Eosinophila showed highly significant difference between atopic dermatitis cases and controls (p < or = 0.001). PMID- 18431990 TI - Contribution of electron microscopic studies to the biology and classification of parasitic cestodes (review article). AB - Members of phylum Platyhelminthes are leaf-like or tape-like worms. Class Cestoidea are exclusively parasitic organisms; covered with a nonciliated integument; ciliated epithelium, when present, confined to embryos (onchospheres) hatched from eggs; scolex provided with suckers and frequently hooks for attachment to host tissue; body in most species divided into separate, sexually complete proglottids. Class: Cestoidea: comprises two subclasses: Cestodaria body undivided; only one set of reproductive organs; oncosphere hatching from egg has 10 hooklets. Cestoda: body typically with scolex and series of proglottids, each with one set (rarely 2 sets) of male and female organs; oncosphere typically has 6 hooklets (Beaver et al., 1984). PMID- 18431991 TI - Denver peritoneovenous shunt in the management of refractory ascites due to chronic liver diseases: impact of patients selection on its outcome. AB - Forty four patients with refractory ascites due to chronic liver diseases that fulfilling the inclusion criteria of selection were divided into 2 groups. The first group (GI, n=24) was subdivided into 2 subgroups according to degree of liver condition; GIa (n=11) with Child-Pugh class B and GIb (n=13) with early class C. The patients were subjected to P-V shunt (Denver group). Similarly, patients in the second group (GII, n=20) were divided into 2 subgroups GIIa (n=10) & GIIb (n=10) respectively and treated by the repeated tapping and albumin infusion (control group). Postoperative results revealed a significant increase in urine out put (P<0.001), decrease in abdominal girth (P<0.01) and body weight (p<0.01) with more patients fitness and satisfaction than in controls. Postoperative complications were more in GIb. Ascites recurrence occurred in 3 (23%) patients in GIb due to severe infection (2 cases) and irreversible shunt obstruction (1 case) and without recurrence in GIa. So, Denver P-V shunt offers a good palliation in such patients, but its use is more justified in selected cases. PMID- 18431992 TI - Impact of experimental duel infections with Schistosoma mansoni and Echinoccocus granulosus on hepatic histopathology. AB - Experimental duel infection with S. mansoni and E. granulosus was induced in mice to determine their effect on serum nitric oxide (NO) level and accordingly on the sequences of histopathological lesions affecting the liver. The results showed that serum NO level was significantly increased (p<0.05) in mice infected with both parasites (GI) in comparison to either S. mansoni (GIV) or E. granulosus (GV). The NO elevation on hepatic pathological lesions of both diseases showed a marked reduction of granuloma size with absence of concentric fibrosis in GI as early as 4 weeks of concomitant infection as compared to GIV. In spite of the significant increase of NO level when E. granulosus infection induced in late stages of schistosomisais (GsII & III), yet granuloma size was not suppressed. Also, there was absence or death of hydatid cyst in mice (GI) compared to E. granulosus (GV). So, the duel infection with the two parasites affected serum NO level and hepatic histopathology, by ameliorative or deteriorative effects, according to duration of infection with either. PMID- 18431993 TI - Mesh repair of non-complicated umbilical hernia in ascitic patients with liver cirrhosis. AB - The policy of elective repair of umbilical hernia in cirrhotic ascitic patients has long been a subject of debate and is still a major health problem. This study evaluated the role and outcome of elective mesh repair of umbilical hernia in cirrhotic ascitic patients, compared with the conventional two layers fascial repair technique. Forty cases with a small to medium sized umbilical hernia defects in ascitic cirrhotic patients were divided into two groups (GI & GII) of 20 patients each. After proper control of ascites in both groups, patients in GI were subjected to elective umbilical hernia onlay mesh repair. In GII, hernias were managed by conventional two layers fascial repair. In GI, transient early postoperative ascitic fluid leakage occurred in 3 (15%) cases and mild superficial wound infection in 5 (25%) cases which was controlled by antibiotics. Two (10%) cases developed hernia recurrence during follow up period. In GII, ascitic leakage occurred in 6 (30%) cases that responded well to conservative management except only one case needed re-exploration and repair reenforcement. Drainage was significantly less than in GI in amount (P<0.05) and duration (P<0.01), wound infection occurred in 3 (15%) cases but were superficial and controlled conservatively. Recurrence occurred in 7 cases (35%). Elective mesh repair of umbilical hernia with ascites in cirrhotic patients proved to be a safe, simple and effective technique with perioperative antibiotics and proper aseptic procedure. PMID- 18431994 TI - Experimental transmission of Hepatozoon gracilis (Wenyon, 1909) com. nov., in its natural host the bean skink lizard (Mabuya quinquetaeniata quinquetaeniata) and vector Culex (C.) pipiens (Diptera: Culicidae). AB - The bean skink lizard Mabuya quinquetaeniata quinquetaeniata (Family: Scincidae) collected from Abu-Rawash, Giza was infected with the protozoon Hepatozoon gracilis. The mean percent of infection rate all over the year was 12.57% with the highest peak was in October (33%), meanwhile the lowest infection rate was detected in June (2.1%) and no infection was detected in February. Microscopical examination of experimentally infected Culex (C.) pipiens L smears revealed the presence of gamogony and sporogony stages of H. gracilis in their haemoceal. Cx. pipiens was capable to transmit H. gracilis to non-infected M. q. quinquetaeniata under laboratory conditions (21 +/- 1 & 60-70% R.H) after a prepatent period of 32 +/- 1 days from the infective bite. Various developmental schizogony stages were detected both in the lung endothelial cells and liver parenchymal cells. Developmental stages of H. gracilis in both vector or vertebrate host were described. Cx. pipiens was unable to transmit H. gracilis to snake, Psammophis schokari present in the same area of the bean skink lizard. PMID- 18431995 TI - [Isolation and identification in Senegal of the most immunogenic protein soluble antigen of a Clostridium chauvoei strain]. AB - Clostridium chauvoei is the pathogenic agent for blackleg, a toxinfection disease in bovine and small ruminants, always lethal and involving considerable economic losses. Some bacteriological, biochemical, immunological studies permitted to isolate identify the major soluble antigenic protein of this bacteria. It's a protein fragment of 70 kDa weight, the 19 fraction, excreted by the bacteria in a suitable culture medium. The 19 fraction of extracellular medium leads to antibodies production on guinea pigs revealed by the ELISA/antibody test. PMID- 18431996 TI - [Disseminated cutaneous histoplasmosis due to Histoplasma capsulatum var. duboisii associated with AIDS. A case report in Abidjan, Cote d'Ivoire]. AB - Histoplasmosis is a deep mycosis whose capsulatum variety is recognized as being an AIDS-defining infection. However AIDS associated with Histoplasma capsulatum var. duboisii is rarely reported. We report a case of cutaneous duboisii histoplasmosis associated with AIDS which has been mistaken for molluscum contagiosum for many months. The diagnosis has been confirmed by means of a biopsy of a nodule followed by an anatomo-pathological examination. The medical treatment was successfully based on combination Triomune (stavudine + lamividine + nevirapine) and itraconazole. PMID- 18431997 TI - [Ocular thelaziasis in man in Northern Italy]. AB - In the present article the author reports a case of ocular thelaziasis in a man living in Piedmont (northern Italy). The subject first complained of symptoms of hyperlacrimation and conjunctivitis. After a series of treatments with eye drops, the nematode responsible for the symptomatology was identified, first mistaken with conjunctivitis. These parasites are rather rare in Italy especially in the North of the country. PMID- 18431998 TI - [Polyneuropathies in patients treated with HAART in Bobo-Dioulasso hospital, Burkina Faso]. AB - Peripheral neuropathies (PN) represent the most common neurological manifestation in patients with HIV infection. Introduction of highly active antiretroviral therapy (HAART) had a significant impact on the epidemiology of HIV-associated neuropathies even in poor-resources countries. HIV-infected patients were followed up over a 2-years period from January 2002 to December 2003. PN was clinically diagnosed based on abnormalities of ankle reflexes or vibratory perception and if patients described pain, paresthesia or numbness. Electromyography was not performed in this study Among the 133 HIV-infected patients treated with HAART 31 patients (23 females and 8 males) with 38.8 of mean age were followed up for PN. 95.5% among them were HIV1-infected. According to the availability of the antiretroviral therapy, 9 patients were treated with protocol A including lamivudine + stavudine + nevirapine, 12 patients with protocol B including combination of stavudine + lamivudine + efavirenz, and 10 patients with protocol C with other combinations of antiretroviral therapies. Average CD4 cell count was 229.3/microl and 60% of the sample had < 200 CD4 cell counts at the time of diagnosis. PN occurred within 5.6 months from the institution of the HAART and 80% less than 3 months after the beginning of the treatment. Burning feet syndrome was found in 16.1% of the sample. 45.2% of polyneuropathies occurred in late stage of HIV infection (< 200 CD4/microl). The presence of PN was related to decreased CD4 cells counts and neurotoxic antiretroviral therapy Introduction of HAART has modified the course and the prognosis of HIV infection even in poor resources setting. The incidence of toxic neuropathies is increasing with longer patients' life expectancy and represents a major factor in treatment limitation and the neurological side effects of HAART should be well identified by physicians. PMID- 18431999 TI - [Conidiobolomycosis, a rare fungal tumor: a case report in Ouagadougou, Burkina Faso]. AB - Conidiobolomycosis is a deep, rare mycosis, due to Conidiobolus coronatus which is a saprophyte of vegetation in decomposition. We report one case in Burkina Faso. A 17 years old man, shepherd, consulted for tumefactions on the face. It could date back insidiously to a traumatism, one year before. A month later some painless tumefactions appeared on the cheekbone, the right eyelid, the nose with epistaxis. The upper lip then the lower one had swollen. Dermatological exam revealed multiple, painless, hard, sub-cutaneous swellings, affecting the cheekbone, the eyelids, the root and the ala of the nose. This tumefaction sometimes adhered to underlying tissues and to the overlying skin, sometimes mobile; painless and hard swelling of the two lips was also noted. ENT exam showed an inflammation of the nasal mucous without ulceration and the permeability of the nasal tracts was subnormal. The cephalic tomodensitometry showed a thickness of the soft tissues of the lips and the nose with an infectious feature associated to a pansinusitis without bone lesion. Histology was in favour of conidiobolomycosis. The patient was treated with fluconazole and the swelling progressively disappeared. Conidiobolomycosis is a disease generally reported in some humid tropical countries. It begins in the nasal cavities leading then to a nasal obstruction. This case was singular by the fact it happened in a dry Sudano-Sahelian climate and by its clinical features. PMID- 18432000 TI - [Disseminated subcutaneous cysticercosis: a report of six cases in Burkina Faso]. AB - Cysticercosis is an infection resulting from the larval form of the pig tapeworm, Taenia solium. The subcutaneous localizations are frequent and can have serious consequences such as neurological attacks. We report six cases among whom five men and a woman, in order to point out the severity of the disease and its possible dissemination. The patients' age was ranging from 25 to 57 years old. Three of them had neurological complications as convulsions and headaches. The nodules were painful in one case. We recommend sanitary education to eradicate the affection and to sensitize patients in order to consult physicians at early stage. PMID- 18432001 TI - [Cerebral malaria in adults at the Infectious Diseases Clinic in the Fann Hospital in Dakar, Senegal]. AB - This study aimed at describing cerebral malaria cases findings in the Fann Hospital in Dakar. Data were collected from patients files recorded from 2001 to 2005. One hundred and twenty nine cases of cerebral malaria were admitted to the clinic, accounting for 21.4% of all malaria cases. The sex-ratio M/F was 2.48 and the mean age of patients 28.24 years old +/- 13.7 [12-85 years old]. Patients presented with either coma (91.4%) or mental confusion (10.07%) along with fever (80.6%), convulsions (33.3%). Other severe malaria conditions were observed: jaundice (7 cases), severe anaemia (5 cases), acute renal failure (3 cases), and circulatory collapse (3 cases). Acute pulmonary infection (4 cases) and Salmonella bacteraemia (2 cases) occurred as complications during patient's hospitalisation. The case fatality rate was 20.2% (26 deaths). No neurological sequelae were found among survivors. Cerebral malaria lethality is still high enough to urge for the improvement of working conditions in our clinic. Together with promotion of preventive measures in the community better health care services will help to reduce malaria related morbidity and mortality. PMID- 18432002 TI - [Gastric cryptosporidiosis revealing a small cell lung carcinoma (Tunisia)]. AB - Cryptosporidium, agent of cryptosporidiosis, is an ubiquitous protozoan organism causing diarrhoea especially in severe immunosuppressed patients. Cryptosporidium has been detected with increasing frequency in the gastrointestinal tract, but involvement of the stomach is rarely reported and discloses an underlying immunodeficiency state. We report the case of 67-year-old man, a heavy smoker, who presented with a history of epigastric pain with an altered general condition. Upper gastrointestinal endoscopy showed no significant mucosal abnormalities. The biopsy revealed a chronic active gastritis with Cryptosporidium parasites lining cryptic epithelium. Systematic chest X ray showed a right suspect parenchymatous opacity. Bronchoscopy with multiple biopsies concluded to a small cell lung carcinoma. Through this rare initial manifestation of immunocompromised state related to cancer we will discuss the role of gastrointestinal endoscopy with biopsies in the diagnosis of cryptosporidiosis. PMID- 18432003 TI - [Anaemia, iron index status and acute phase proteins in malaria (Abidjan, Cote d'Ivoire)]. AB - Clinical signs of malaria are the combined expression of several biological mechanisms. During this parasite infection, anaemia can be the consequence of several different pathogenic mechanisms. It can be an acute haemolytic anaemia due to a mechanical and immune action of the parasite or an inflammation. Besides, in Africa malaria matches with iron deficiency area. So, malarial anaemia in tropical area can be a characteristic of iron deficiency The purpose of this survey was to define the features of malarial anaemia and elucidate the link of all biological processes involved. A black population living in tropical urban areas, with fever and diagnosed Plasmodium-infection was assessed. Parasitaemia, haemoglobin, hematocrit, average corpuscular volume and average corpuscular haemoglobin were determined. For each patient, iron index status and acute phase protein were assessed with the plasmatic iron, ferritin, haptoglobin, transferrin and C-reactive protein. Regardless of gender and age, the characteristics of malarial anaemia are microcythaemia and hypochromia. Anaemia occurs as frequently as parasitaemia is high. When parasitaemia is low anaemia gets a haemolytic feature. When parasitaemia is high, anaemia gets haemolytic and inflammatory features. Anaemia occurs more often with a good iron index status. PMID- 18432004 TI - [Cutaneous leishmaniasis caused by Leishmania infantum MON-24 in Tunisia: extension of the focus to the center of the country]. AB - Three clinico-epidemiological forms of cutaneous leishmaniasis (CL) were described in Tunisia: the zoonotic CL (ZCL) epidemic which occurred in the centre of the country caused by Leishmania major MON-25, the chronic CL (CCL) In the south-east of the country caused by Leishmania killicki MON-8 and the sporadic CL In the North (SCL) caused by Leishmania infantum MON-24. The latter form, described in 1991, prevails in northern Tunisia with approximately thirty cases per year. Its vector, unknown for a long time could be according to the last publications, Phlebotomus perfiliewi or Phlebotomus langeroni; however, its reservoir remains unknown until now. The systematic isoenzymatic characterization permits to identify a great number of strains improving then knowledge on the eco epidemiology of the disease. Indeed, changes were noted in the geographical distribution of these clinical forms: extension of the ZCL to the North and South, extension of the CCL to North and the SCL to the centre. We report in this note the first mention of L. infantum MON-24 in the two provinces of the centre of Tunisia: Kairouan and Sidi Bouzid, confirming the extension of the SCL to the Centre. PMID- 18432005 TI - [Delayed home screening of leprosy; experience of the screening team in Mayotte]. AB - Mayotte, a French territory island located in the Indian Ocean near Madagascar, remains a leprosy endemic area. In 2006, leprosy was still a problem of public health with a prevalence of 3.94 per 10,000 inhabitants. There is practically no formal consensus about active screening (AS) on an index case. According to teams and their related staffs, the AS concerns intradomicilary contact individuals (IDC) restrictively or extended to extra-domicilary social and professional contacts. Date, number and frequency of these investigations depend on each team. Between 1997 and 2003, there was no AS planned in Mayotte, but all index case individuals have been encouraged to propose a screening to their relatives through specific campaign information and education. This procedure allowed to identify 10 new cases of leprosy infection among the IDC. Concurrently 12 IDC cases have been diagnosed by health workers. In 2003, we performed a postponed AS within IDC of every Mahorais case registered by passive detection between 1997 and 2003. 325 IDC have been examined and 15 new cases have been detected. All these new cases showed early leprosy features: 14 were paucibacillary forms, among which 9 cases with an isolated cutaneous lesion (7 had an infracentimetric lesion). One patient had multibacillary disease although he presented with an isolated skin lesion which developed within the 6 previous months. None presented with disability. Our results suggest that passive detection even reinforced by repeated individual information and education about leprosy is neither appropriate nor effective. The postponed AS seems to favour an increased self esteem and a better involvement of the index patient in sanitary education together with the screening of his relatives. In the Mayotte background, the postponed AS has not been associated with a significant delay for diagnosis. Although WHO recommandations are to abandon immediate AS of IDC and to promote self-screening for leprosy our study suggests an intermediate position, namely delayed active screening for an enhanced effective detection. PMID- 18432006 TI - [Serological study carried out in Cambodia during a tetanus vaccination in adults]. AB - In 1997, the Ministry of Health tested the feasibility and serological activity of a two-dose vaccine at one year interval within a catch-up tetanus immunization programme in a rural population. In the district of Angkor Thom in the Siem Reap province (15,000 inhabitants), a team of nurses and administrative clerks travelling by motorcycle, conducted between February 1998 and February 1999 an EPI and tetanus immunization of the whole population gathered in meeting points. In 132 childbearing age female volunteers, 49 following a two-dose schedule at one year interval, and 70 following a WHO three-dose schedule, with two doses at one month interval and a booster dose one year later tetanus antibodies have been measured before vaccination, one year after the first dose or the two first doses, and six months after the second or third dose of vaccine. 129 male volunteers of the same age were also recruited in the serological study following only the two-dose schedule. The titration was done first with monoantigen ELISA, then with mouse seroneutralisation, the reference method for measuring tetanus seroprotection. Only 148 (57%) volunteers completely attended the serological study Compared to seroneutralisation, sensitivity for seroprotection with ELISA was 89% (CI95%: 85%-94%) and specificity 84% (CI95%: 81%-89%). The coverage of the general population vaccinated with two doses, in both sexes and in all age groups, increased on average from 5% to 70%. The three-dose schedule gave significantly more protection than the two-dose schedule in women tested with seroneutralisation. On a first sample in those with no protective antibodies and testifying they had not been vaccinated before, 51% of these volunteers after one dose and 93% after two doses acquired protective antibodies. On first sample, 52% of female volunteers had protective antibodies in seroneutralisation, against 11.7% in men. 14% of subjects tested in ELISA and 6.8% tested in seroneutralisation showed in a second sample a decrease in titres, although they had received a tetanus vaccine. For unprotected volunteers on first sample and testifying they had not been vaccinated before, neither age nor past chronic cutaneous lesions or cows living around their houses, two possible sources of contact with CI. tetani, increased significantly seroconversion. Only female volunteers were significantly more seroconverting (81%) compared to men of same age (51%) (RR: 1.60, CI95%: I. 17-2.18) suggesting a memory bias in women supposed to be vaccinated by EPI. 30% of volunteers in ELISA and 14% in seroneutralisation showed spontaneous protecting antibodies in the first sample without having any document or memory of a past tetanus vaccination. Tested by seroneutralisation, no relation was to be found for having spontaneous antibodies with past chronic cutaneous lesions and cows living around their houses. Only the eldest (35-45 y.o.) female volunteers showed significantly more spontaneous antibodies (RR: 3.83, CIs%: 1.74-8.2) than men in the same age-group. A memory bias may be found also in this female age-group. Good serological response should encourage implementation of a catch-up tetanus vaccination in this country considering the large number of unprotected adults, mainly male adults. Due to problems with notification and recalling past vaccinations, only a prospective study in an unimmunized large cohort, studying all possible factors of tetanus toxin neutralisation, could confirm the existence and cause of spontaneous antibodies. Excluding vaccination in at-risk population for such a study would be however ethically unacceptable. PMID- 18432007 TI - [Systematic controlled placenta birth: analysis of 200 cases recruited at the general hospital of Abobo (Abidjan, Cote d'Ivoire)]. AB - Haemorrhage of placenta birth is one of the main reasons of maternal mortality in developing countries. The control of this third stage of delivery remains one of the means to stem the scourge. This survey shows that controlled placenta birth is a safe mean to reach this objective: 200 women who had normal vaginal deliveries at the Abobo North hospital were included in the survey. Among them, one hundred women had a controlled placenta birth whereas 100 had a normal one. The analysis of the results showed that: the haemorrhage rates during placenta birth slightly decrease in the group with controlled placenta birth and are clearly lower in the group of patients with risks factors of haemorrhage in 10% of the cases; the delay of placenta birth is twice shorter in the group having a controlled placenta birth than in the group with normal placenta birth; blood loss in the group with controlled placenta birth is three times less important than in the other group. This study speaks in favour of a systematic controlled placenta birth during the third stage of labour as it is already performed elsewhere. PMID- 18432008 TI - [Plasmodium falciparum chloroquine and pyrimethamine resistance monitoring network with molecular tools in the Niger River valley, Republic of Niger]. AB - Plasmodium falciparum resistance to chloroquine first arose in Africa 25 years ago. Nowadays most of African malaria control programmes have switched their first-line treatment of uncomplicated malaria cases towards artemisinin derivatives combination. After WHO guidelines, a survey network for malaria treatment resistance has been set up in the Niger valley around Niamey since December 2004. The association of the Niger national malaria control programme with the CERMES research center allowed collecting of samples from both health centers and hospitals of this region. Blood finger-pricks on filter papers were tested for detection of plasmodial antigen in health center without biological diagnosis capacity. Specimens found positive either in hospital laboratory or by using antigen method were tested by PCR/RFLP to detect K76T mutations on the pfcrt gene and S108N mutation on the pfdhfr gene. This simple procedure allows the screening of a large number of specimens. Moreover, a spatial distribution of mutations and evidence of resistance clusters were searched integrating the data in a geographic information system. The 76T mutation of pfcrt and 108N of pfdhfr were respectively found in 50.8% and 57% of the specimens tested. No statistically significant difference was found according to the level of sanitary formations or the age of the patients. No resistance cluster was identified and the prevalence of mutation seems homogeneous in the zone. By completing the clinical efficacy studies we think that our simple method for collecting and testing blood samples associated with clinical efficacy studies may be useful for building a network of malaria drug resistance in Africa. PMID- 18432009 TI - [Symptomatic and asymptomatic Plasmodium falciparum infection in children from 6 months to 6 years old in the Abobo general hospital (Abidjan, Cote d'Ivoire)]. AB - It is commonly admitted that people living in malarial zone are carrying asymptomatic Plasmodium. Cote d'Ivoire is one of these zones. The studies carried out on malaria in these areas have focused mainly on the clinical forms of the disease and effectiveness of the antimalarial drugs. The purpose of this study was to determine the prevalence of the symptomatic and asymptomatic carriage of Plasmodium falciparum in children of 6 months to 180 months old in the Abidjan area. Over a period of twelve months, 902 feverish subjects and 681 non-feverish subjects were selected among the 7,017 people admitted in the paediatrics service of the Abobo general hospital for detection of malaria parasite. Among 1,583 selected subjects, 358 were carrying Plasmodium falciparum implying a total prevalence rate of 22.6%. The prevalence rate was 13.5% and 29.5% respectively in the asymptomatic subjects and symptomatic subjects. The highest proportions of positive thick smears were observed during the long rainy and dry seasons but, parasitaemia was the highest during the short dry season. In 31.5% of the cases, the asymptomatic carriers had a parasitic density higher or equal to 10,000 trophozoites/microl of blood and fever was not related to parasitic load. The prevalence rates of Plasmodium carriage and malaria were higher during the long rainy season. This study highlighted a considerable proportion of asymptomatic Plasmodium falciparum carriers. Improving environmental conditions should help to reduce this rate of carriage. PMID- 18432010 TI - [Post-circumcision tetanus in Dakar, Senegal]. AB - This study aimed at describing the epidemiology, clinical features and prognosis of post-circumcision tetanus at the infectious diseases clinic in Fann Hospital in Dakar. Data were collected retrospectively for analysis from patients' files recorded from January 1, 1999 to December 31, 2006. 54 cases were included, accounting for 4% of all tetanus cases admitted to the clinic during the study period (54 cases/1291). The patients' average age was 9 +/- 3.7 years old (range = 1-17 years) and 52% of them were schoolboys. In most cases (76%), tetanus symptoms occurred beyond 7 days after circumcision. The average delay from onset of the disease to admission was 2.3 days (range = 0-6 days). The circumcision took place at home in 39% of cases, in health center in 35% of cases and in unspecified area in 26% of cases. The majority of patients (85%) had never received tetanus vaccine and, in 72% of the cases, the circumciser was designated as a male nurse. Generalized tetanus was observed in all cases, most of which was a mild form of the disease (63%). During hospitalisation, thirteen patients (24%) had complications among which diaphragmatic and intercostal muscle spasms (3 cases), bacteraemia (5 cases), respiratory infection (4 cases), urinary tract infection (4 cases), and fracture of the vertebrae (1 case). The case fatality rate was 7.4% (4 deaths). Vaccination together with health education of the population as well as a better sensitization of the practitioners are necessary to eradicate tetanus after circumcision. PMID- 18432011 TI - [Leprosy and immune reconstitution syndrome in AIDS]. AB - The immune reconstitution syndrome (IRS) has been typical of changes in the clinical presentations of opportunistic infections in AIDS patients since the introduction of HAART. Leprosy has joined the growing list of opportunistic infections associated with IRS. PMID- 18432012 TI - [Three new cases of visceral leishmaniasis in Cote d'Ivoire]. AB - The objective of this work is to report three cases of visceral leishmaniasis in non-HIV infected native patients in Cote d'Ivoire. The three observations concerned adults aged of 31 and 65 and a five years old child without particular medical or surgery histories. Factors associated with visceral leishmaniasis regarding the younger and the older adults were respectively young age, chronic lymphoid leukaemia and Burkitt lymphoma. Clinical features in the three cases were chronic fever a myeloproliferative syndrome with wasting syndrome and pancytopenia. The diagnosis was confirmed by the existence of Leishmania in bone marrow, ganglionic juice and splenic samples. The species of Leishmania was not identified. Specific treatment was based on intravenous amphotericin B (Fungizone) relayed by Glucantim in one case because of side effects; however the treatment has been unsuccessful. These three new cases show that visceral leishmaniasis is a reality in Cote d'Ivoire therefore an epidemiological survey is requested in order to identify Leishmania species, reservoirs and vectors. PMID- 18432014 TI - Well equipped for the new year. PMID- 18432013 TI - [Extension to the north of Morocco of the distribution area of Anopheles (Cellia) d'thali Patton, 1905]. AB - Anopheles (cellia) d'thali is generally classified as a mosquito of arid areas in the South and East Morocco. The northernmost station of this species at present in Morocco is the Moulouya valley. However we found An. d'thali during entomological investigations in the north of the country in the subhumid area of Chefchaouen. In Morocco, An. d'thali is therefore no longer a strictly desert species. PMID- 18432015 TI - Comparison of the static loading capacity of all-ceramic bridge frameworks in posterior teeth using three hard core materials. AB - In this in vitro investigation, we studied the static strength of three-unit all ceramic bridge frameworks with rigid positioning of the abutments. The materials used were Vita InCeram Alumina, Vita InCeram Zirkonia, and zirconium oxide (Vita InCeram (X YZ Cubes for Cerec). A finite element calculation was performed for numerical comparison of the load-bearing capacity of two main normal tensions. The purpose of the investigation was to design a framework made from hard core material, that offers the greatest possible resistance in static fracture loading tests. In a model, the 2nd maxillary right premolar and 2nd maxillary right molar served as bridge abutments and were provided with a chamfer preparation. On this base, two different bridge frameworks were constructed using CAD/CAM technology after an impression had been taken. One bridge connector was designed heart shaped, with contact to the gingiva, while the other was designed as a "free connector" at a distance of 1.2 mm from the gingiva. In this framework design, the radius in the cervical connector area is larger. We were uncertain as to whether it would be possible to further increase the strength of the ceramic material by the use of the differently designed pontic, independent of the hard core ceramic used. The least fracture strength was registered for the "heart shaped connector" constructed from InCeram Alumina, with a mean fracture load of about 1089 Newton (N). The connector designed as a "free connector", made from the same material, was stronger by 10%. With the materials InCeram Zirkonia and zirconium dioxide as well, the "free-connector" design achieved a 10% higher breakage limit than the heart-shaped design. InCeram Zirkonia was 25% more stable in the static load tests than InCeram Alumina. Zirconium dioxide demonstrated a 2.3-fold greater strength than InCeram Alumina, while the free-connector design showed the greatest mean static loading capacity of 2808 N. PMID- 18432017 TI - Risks in the digital transmission of sensitive patient data. AB - Currently, methods of modern imaging are increasingly assuming a predominant position in diagnostics and therapy. Larger and larger amounts of sensitive data and information are generated, processed and--frequently with inadequate protection--transmitted through generally accessible networks. The Internet is used especially intensively as a medium for transporting digital data; at the same time, protection of sensitive data is frequently neglected to a high degree by medical professionals. The present contribution is therefore intended to provide an overview of currently valid standards and legal regulations, as well as suitable encoding methods relevant for the dental practice. PMID- 18432016 TI - Fit of surgical guides--manufactured by InLab 3D. AB - Based on an exclusively virtual implant planning and positioning concept, the fit of surgical-guide-like workpieces was evaluated in an in-vitro pilot study, manufactured with the InLab 3D system. From 85 maxillary casts, 8 were randomly selected. The first right maxillary incisor was removed. After optical impression of the maxillary anterior teeth, a drilling template was designed by the use of the InLab software, version 3.0. The mode "bridge" "dental database" was applied. The guidance was milled from a methacrylate block and braced on the adjacent teeth. After milling, they were checked for proper seat (without rocking). The discrepancies between the inner surface of the template and the opposing tooth surface was shown with a silcone based material, which rested on the casts after removing the template. The difference between the thickness of the silicone layer and the subjacent tooth surface was measured with a high resolution scanner. In order to characterize the fit of the 8 workpieces the average mean, average maximum and average minimum values of the fit were calculated in the buccal, incisal and palatinal area using the program Match 3D (W. Gloger, LMU): average mean: buccal: 65 microm; incisal: 116 microm; palatinal: 94 microm; average maximum: buccal: 162 microm; incisal: 202 microm; palatinal:190 microm; average minimum: buccal: 5 microm; incisal: 14 microm; palatinal: 15 microm. In the present study it was possible to produce drilling templates with the already commercially available InLab system. The results were presented at the Annual Meeting of the German Society of Computerized Dentistry 2007 as a short presentation. PMID- 18432018 TI - The ISCD section of the FDI congress in Dubai. PMID- 18432019 TI - Suspected radioactive contamination: evaluation of 45 Israeli citizens potentially exposed to polonium-210 in London. AB - The lethal poisoning of Alexander Litvinenco with the radioactive element polonium-210, and the risk that many civilians (including Israeli citizens) who were in the same location in London at the same time were exposed to radiation, was an unprecedented event in the western world. This was only the second known death due to 210Po, a natural alpha radiation-emitting element. A task team was created to handle the event. The team comprised representatives from the Ministry of Health's advisory committee for radiological events (which includes the Israel Defense Force, the Israeli Atomic Energy Commission and the Ministry of Environmental Protection), the Public Health Services Central District, and a public relations expert. Forty-seven people were located and underwent an epidemiological inquiry, and urine samples for detection of 210Po were sent abroad to a specialized laboratory. The radiotoxicological results were analyzed and evaluated by the expert team and follow-up recommendations were made. This unfamiliar and potentially stressful scenario was handled successfully by a multi organizational multidisciplinary task team. The joint work of the task team was a real-life "exercise" simulating a radiological event in Israel. This team has recommended further evaluation of various vital missions in the event of any possible future radiological event, with special emphasis on a proactive communication approach to the media and the public. PMID- 18432020 TI - Admission for syncope: evaluation, cost and prognosis according to etiology. AB - BACKGROUND: Syncope is a common clinical problem that often remains undiagnosed despite extensive and expensive diagnostic evaluation. OBJECTIVES: To assess the diagnostic evaluation, costs and prognosis of patients hospitalized for syncope in a tertiary referral center according to discharge diagnosis. METHODS: We retrospectively reviewed the medical records of patients with a diagnosis of syncope discharged from a tertiary referral center in 1999. In addition, mortality data were obtained retrospectively a year after discharge for each patient. RESULTS: The study group comprised 376 patients. Discharge etiologies were as follows: vasovagal 26.6%, cardiac 17.3%, neurological 4.3%, metabolic 0.5%, unexplained 47.3%, and other 4%. A total of 345 patients were admitted to the internal medicine department, 28 to the intensive cardiac care unit, and 3 to the neurology department. Cardiac and neurological tests were performed more often than other tests, with a higher yield in patients with cardiac and neurological etiologies respectively. The mean evaluation cost was 11,210 +/- 8133 NIS, and was higher in the ICCU than in internal medicine wards (19,210 +/- 11,855 vs. 10,443 +/- 7314 NIS, respectively; P = 0.0015). Mean in-hospital stay was 4.9 +/- 4.2 days, which was longer in the ICCU than in medicine wards (7.2 +/ 5.6 vs. 4.6 +/- 3.5 days, respectively; P = 0.024). Short-term mortality rates (30 days after discharge) and long-term mortality rates (1 year after discharge) were 1.9% and 8.8% respectively, and differed according to discharge etiology. LTM rates were significantly higher in patients discharged with cardiac, neurological and unknown etiologies (not for vasovagal), compared with the general population of Israel (1 year mortality rate for the age-adjusted [65 years] general population = 2.2%). The LTM rate was higher in patients discharged with a cardiac etiology than in those with a non-cardiac etiology (15.4% vs. 7.4%, P = 0.04). Higher short and long-term mortality rates were associated with higher evaluation costs. CONCLUSIONS: Hospitalization in a tertiary referral center for syncope is associated with increased mortality for most etiologies (except vasovagal), cardiac more than non-cardiac. Despite high costs of inpatient evaluation, associated with more diagnostic tests, longer in-hospital stay and higher mortality rates, nearly half of the patients were discharged undiagnosed. Outpatient evaluation should be considered when medically possible. PMID- 18432021 TI - Usefulness and predictive value of circulating NT-proBNP levels to stratify patients for referral and priority treatment in a specialized outpatient heart failure center. AB - BACKGROUND: Serum natriuretic peptide levels are useful diagnostic and prognostic markers in patients with acute decompensated heart failure, but have been little used to stratify urgency of treatment in the outpatient situation. OBJECTIVES: To examine the use of natriuretic peptide to guide priority of patient referral to a heart failure center. METHODS: We analyzed data from 70 consecutive patients with chronic heart failure (NYHA class 2-4) referred for first evaluation in a specialized outpatient heart failure center. Serum NT-proBNP was measured at the initial patient visit. We examined correlates and predictive value of mid- and upper tertile NT-proBNP for mortality in comparison with other known prognostic indicators using univariate and multivariate logistic regression analysis. RESULTS: Mortality at 6 months was 26.0% in patients with upper tertile (> 1958 pg/ml) NT-proBNP, 8.7% in the middle tertile group and 0% in the lowest tertile (P=0.017). Patients with upper tertile serum NT-proBNP levels (group 3) had lower left ventricular ejection fraction, were more often in atrial fibrillation (P=0.04) and more often had renal failure (P=0.03). Age-adjusted logistic regression analysis identified upper tertile serum NT-proBNP level as the strongest independent predictor of 6 month mortality with a sixfold risk of early death (adjusted odds ratio 6.08, 95% confidence interval 1.58-47.13, P=0.04). NT proBNP was a more powerful predictor of prognosis than ejection fraction and other traditional outcome markers. CONCLUSIONS: In heart failure patients referred to an outpatient specialized heart failure center, an upper tertile NT proBNP level identified patients at high risk for mortality. A single high > 550 pg/ml NT-proBNP measurement appears to be useful for selecting patients for care in a heart failure center, and a level > 2000 pg/ml for assigning patients to high priority management. PMID- 18432022 TI - Improved exercise capacity in patients after minor ischemic stroke undergoing a supervised exercise training program. AB - BACKGROUND: Regular physical activity is known to have a beneficial impact on multiple cardiovascular risk factors, but there is no routine provision of exercise training programs for patients after ischemic stroke. OBJECTIVES: To assess the tolerability, safety and effect of an outpatient supervised exercise training program in patients after a non-disabling ischemic stroke. METHODS: Patients discharged home following a minor ischemic stroke (modified Rankin scale; mRS < or = 2) were referred to a 3 month outpatient supervised exercise training program, performed twice weekly as prescribed by a physiologist and supervised by physical therapy. Exercise capacity was evaluated by the 6 minute walk test and the modified Bruce exercise test. RESULTS: Of the 52 patients who met the selection criteria, 43 underwent supervised exercise training within 2 months of stroke onset and 9 did not (control group). The baseline characteristics were comparable between the two groups. Following the exercise training program, an improvement in exercise capacity was observed manifested by improvement in the 6 minute walk test (444 +/- 90 to 557 +/- 99 meters in the exercise group vs. 438 +/- 101 to 418 +/- 126 in the control group; P = 0.002 for the score changes) and in the exercise duration achieved in the modified Bruce test and the metabolic equivalents achieved [9.6 +/- 3.7 to 12.4 +/- 3.2 minutes and 6.2 +/- 2.8 to 8.5 +/- 3.4 respectively in the exercise group (n = 41) vs. 9.2 +/- 3.5 to 8.0 +/- 3.4 min and 5.8 +/- 1.8 to 5.8 +/- 2.8 in the control group (n = 7); P = 0.0009 and 0.01 for score changes, respectively]. CONCLUSIONS: An outpatient supervised exercise training program after a minor ischemic stroke is feasible, well tolerated and is associated with improvement in exercise capacity. We strongly recommend that an aerobic exercise program be offered to suitable patients after an ischemic stroke. PMID- 18432023 TI - Computed tomography angiography in pulmonary hypertension. AB - BACKGROUND: Diseases causing increased pulmonary pressure will subsequently cause a dilation of the pulmonary arteries and right heart chambers. OBJECTIVES: To assess the capability of computed tomography angiography and high resolution CT to diagnose and estimate the severity of pulmonary arterial hypertension as compared with standard means of right heart catheterization, echocardiography and pulmonary function tests. METHODS: The study included 38 patients with PHT who underwent CT angiography and HRCT as part of their routine evaluation. Diagnoses included: primary PHT (n=20), Eisenmenger syndrome (n=6), scleroderma (n=3), thromboembolic disease (n=3), and others (n=6). Mean pulmonary artery pressure was 58 mmHg (range 39-92 mmHg) by catheterization and peak systolic pressure 79 mmHg (range 40-135) by echocardiography. Findings for the diameters of the main pulmonary artery and its main branches, the ascending aorta, the right atria and ventricle as well as the position of the interventricular septum were compared with 22 chest CT scans of patients with no known clinical history of pulmonary hypertension, performed for other reasons (trauma, oncology follow-up) during the study period. Correlations were also calculated with recent right heart catheterization, echocardiography and pulmonary function tests of the study group. RESULTS: Mean main pulmonary artery diameter in the study group was 3.55 +/- 0.66 cm, pulmonary artery/ascending aorta ratio 1.2 +/- 0.29, right pulmonary artery 2.63 +/- 0.49 cm, left pulmonary artery 2.57 +/- 0.5 cm. All diameters were significantly different from the control group (P < 0.0001). Main and right pulmonary artery diameters correlated with the pressure measurement by echocardiography (P=0.001). Bronchial collaterals were found in 11 patients (30%). The position of the interventricular septum correlated well with the echocardiography study. CONCLUSIONS: The size of the main pulmonary artery on CT angiography has a good predictive value regarding the severity of PHT. PMID- 18432024 TI - Carotid artery stenting in high risk patients with carotid artery stenosis not eligible for endarterectomy: clinical outcome after 5 years. AB - BACKGROUND: Carotid artery stenting is used as an alternative to surgical endarterectomy. OBJECTIVES: To determine the outcome of CAS in a retrospective cohort of patients. METHODS: Between July 1999 and March 2003, 56 consecutive patients with carotid artery stenosis who were considered ineligible for surgery were treated (45 males, 11 females, mean age 69). All underwent the procedure prior to the introduction of distal protective devices in Israel. RESULTS: Intraprocedural complications included transient neurological findings in 5 patients (8%), cerebrovascular accident in 2 (3%), hemodynamic changes in 11 (18%), and 4 procedural failures. Post-procedural complications included transient ischemic attack in 3 patients and cardiovascular accident in 6 (10%). At 30 days follow-up, three patients (5%) remained with signs of CVA. Two patients (3%) died during the post-procedural period and 16 (28%) during the 5 year follow-up, one due to recurrent CVA and the remainder to non-neurological causes. Five-year carotid Doppler follow-up was performed in 25 patients (45%), which revealed normal stent flow in 21 (84%), 50-60% restenosis in 3 (12%) and > 70% restenosis in one patient (4%). CONCLUSIONS: This study confirms that stent procedures are beneficial for symptomatic carotid stenosis in patients not eligible for surgery. PMID- 18432025 TI - A simplified fall-risk assessment tool for patients hospitalized in medical wards. AB - BACKGROUND: Falls are a common problem among hospitalized patients, having a significant impact on quality of life and resource utilization. OBJECTIVES: To develop and validate a fall-risk assessment tool for patients hospitalized in the department of medicine that will combine simplicity with adequate accuracy for routine use. METHODS: This observational cohort study was conducted on the medical wards of an urban tertiary teaching hospital, and included all patients who fell in the medical wards during a 1 year period (n = 140) compared to other hospitalized patients. RESULTS: Significant correlates of falls were previous falls, impairing medical conditions, impaired mobility, and altered mental state. In multivariate logistic regression analyses, only previous falls (odds ratio 3.8 with 95% confidence interval 2.65-5.45, P < 0.0001) and acute impairing medical conditions (OR 1.56, CI 1.06-2.29, P < 0.05) correlated independently with a higher risk for falls. Impaired mobility retained an OR of 1.46 (CI 0.95-2.24, P = 0.084). Accordingly, defining patients with either a history of previous falls or both acute impairing medical state and impaired mobility as fall-prone patients provided a sensitivity and specificity of 67% and 63%, respectively. In a subsequent prospective validation trial on 88 patients who fell during hospitalization and 436 controls, the sensitivity and specificity of this fall risk grouping were 64% and 68% respectively. CONCLUSIONS: Our new simple and easy to-use fall-risk assessment tool identified most of the fall-prone patients. These findings suggest that this tool may enable us to prevent two-thirds of falls on the medical ward by providing effective fall-prevention facilities to only one-third of the patients. PMID- 18432026 TI - New and improved Israeli reference of birth weight, birth length, and head circumference by gestational age: a hospital-based study. AB - BACKGROUND: Many centers in Israel still use pre-1970 reference data for neonatal weight, length and head circumference. A recently published population-based reference overestimated the weight of premature infants. OBJECTIVE: To develop a national reference for birth weight, birth length and head circumference by gestational age for singleton infants in Israel. METHODS: Data were collected on all singleton live births documented in the neonatal registry of Rabin Medical Center from 1991 to 2005 (n=82,066). Gestational age estimation was based on the last menstrual period until 1977 and early fetal ultrasound thereafter. Neonates with an implausible birth weight for gestational age (identified by the rule of median +/- 5 standard deviations or expert clinical opinion) were excluded. Reference tables for fetal growth by gestational age were created for males and females separately. RESULTS: The growth references developed differed markedly from the Usher curves currently used in our department. Compared to the recently published population-based birth weight reference, our data were free of the problem of differential misclassification of birth weight for gestational age for the premature infants and very similar for the other gestational age groups. This finding reinforced the validity of our measurements of birth weight, as well as of birth length and head circumference. CONCLUSIONS: Use of our new (birth length and head circumference) and improved (birth weight) gender-specific hospital based reference for fetal growth may help to define normal and abnormal growth in the neonatal population of Israel and thereby improve neonatal care and public health comparisons. PMID- 18432027 TI - Abdominal stab wounds in Israel, 1997-2004: significant increase in overall incidence and prevalence of severe injury. AB - BACKGROUND: Non-operative management following abdominal stab wounds is possible in selected patients who are both hemodynamically stable and do not have signs of peritonitis. However, the rate of failure of non-operative management is higher in Israel than in western countries. OBJECTIVES: To assess the patterns of injury following abdominal stabbing. METHODS: Data from the Israeli Trauma Registry were used to identify all patients with abdominal stab injury admitted to eight different trauma centers between 1997 and 2004. RESULTS: The number of patients admitted per year more than doubled between 1997 and 2004, from 257 to 599. The percentage of patients with severe injury (Index Severity Score > or = 16) increased from 9.4% to 19.0%. The incidence of multiple stab injuries almost doubled, from 37% to 62%. CONCLUSIONS: Review of the data in the Israeli Trauma Registry indicates an increase in both absolute rate and relative incidence of serious stab injuries. This indicates that patterns of injury following stab wounds are not necessarily similar, not even within the same geographical area over time. PMID- 18432028 TI - Issues in rheumatology and autoimmunity. Preface. PMID- 18432029 TI - Tailor-made therapy in rheumatoid arthritis: fact or fiction? AB - There has been a paradigm shift in the treatment of rheumatoid arthritis in recent years. Early and aggressive treatment with good control of disease activity has improved the prognosis of the disease, however, there is significant variability in the response of patients to different therapeutic agents. Hence it is essential to find the predictors of response to a drug at baseline so that we can avoid the delay in achieving remission and improve the outcome. Here we review the literature on available predictors for treatment response in general and specifically for methotrexate and biological agents. We also look at specific scores or indices that can help predict the response in individual patients. PMID- 18432030 TI - Strychnine--a killer from the past. PMID- 18432031 TI - Non-invasive monitoring of inflammation in asthma using exhaled nitric oxide. AB - Management of asthma is currently based on symptoms (in children, usually a second-hand report from parents) and lung function measurements. Inhaled steroids, targeted at controlling airway inflammation, are the mainstay of asthma management. Due to possible side effects they should be used at the lowest possible doses while asthma is adequately controlled. Fractional exhaled nitric oxide is a simple non-invasive method to assess inflammation in asthma, and its role in asthma management is increasing in popularity. The present review summarizes recent research on the use of FeNO in monitoring airway inflammation and optimizing asthma management. The addition of FeNO measurements to the conventional assessment of asthma control appears promising. The practicability of including this measuring method into everyday clinical practice is currently being evaluated. PMID- 18432032 TI - Syncope: diagnostic challenge and economic burden. PMID- 18432033 TI - B-type natriuretic peptide: a universal cardiac biomarker? PMID- 18432034 TI - Amniotic fluid embolism: a plea for better brain protection. PMID- 18432035 TI - Vasculitis and myelodysplasia. PMID- 18432036 TI - Takayasu's arteritis as a cause of carotidynia: clinical and imaging features. PMID- 18432037 TI - Twiddler's syndrome: a rare cause of pacemaker failure. PMID- 18432038 TI - Lymphangioma of the thyroid. PMID- 18432039 TI - Bacteremia and pyelonephritis caused by Lactobacillus jensenii in a patient with urolithiasis. PMID- 18432040 TI - Doctoral degrees for PAs: what happens next? PMID- 18432041 TI - What caused the rash on this elbow? Papular acrodermatitis of childhood. PMID- 18432042 TI - Colon cancer: an update on screening. PMID- 18432043 TI - How biological abnormalities separate CFS from depression. PMID- 18432044 TI - Brain abscess as a sign of hereditary disease. PMID- 18432045 TI - A serum sickness-like reaction to a commonly used acne drug. PMID- 18432046 TI - A fresh look at continuous subcutaneous insulin infusion. PMID- 18432047 TI - New diagnostic tools for managing the febrile infant. PMID- 18432048 TI - Talking the talk, or what it's like on the other side of the stethoscope. PMID- 18432049 TI - Helping to ensure safer transitions in hospitals. PMID- 18432050 TI - What types of birth control are available? PMID- 18432051 TI - A 50-year-old man with a serious pain in the neck. PMID- 18432052 TI - Case of the month. Gastroesophageal reflux disease (GERD), heart failure, valvular heart disease. PMID- 18432053 TI - Time course of pulmonary pathology, cytokine influx and their correlation on augmentation of antigen challenge by influenza A virus infection. AB - A murine model of influenza A virus exacerbation of allergen induced airway inflammation, pulmonary histopathological changes, bronchoalveolar lavage fluid (BALF) analysis, cytokine influx and the time course of these events have been studied. The present study was undertaken to determine the relative contributions of Thl/Th2 cytokines to the histopathological changes in the lungs observed at 9, 12, 24 and 48 hr following antigen challenge in mice previously immunized with influenza A virus. BALF analysis of acute phase group revealed statistically significant increase in neutrophils at 9 hr, macrophages at 12 hr, lymphocytes and eosinophils at 24 hr, as compared to OVA-sensitized control mice. These changes were associated with an alteration in the levels of IL-4, IL-5 and IFN gamma. A peak of IL-4 at 24 hr significantly enhanced bronchiolar and perivascular histopathology, whereas increased IL-5 level peaking at 24 hr was correlated with the enhanced infiltration of eosinophils in both BALF and lung tissue. There was simultaneous depletion of IL-10 an anti-inflammatory cytokine leading to persistence of pulmonary inflammation in case of acute phase group. Histopathology at 24 and 48 hr showed severe denudation of bronchiolar lining epithelium surrounded by dense chronic inflammatory infiltrate. Chronic interstitial infiltrate with focal loss of architecture, marked oedema, extravasation of RBCs from congested blood vessels and laying down of reticulin fibres was observed in acute phase. Thus, infection with influenza A virus on pre existing asthmatic immunopathology elicits a cascade of Th2 cytokines with influx of inflammatory cells in BALF, mucosal and interstitial inflammation leading to asthma exacerbations. PMID- 18432054 TI - Gabapentin attenuates acute hypoxic stress-induced behavioral alterations and oxidative damage in mice: possible involvement of GABAergic mechanism. AB - The effect of gabapentin has been investigated on acute hypoxic stress-induced behavioral alterations and oxidative damage in mice. Mice were subjected to hypoxia for 2 hr. Treatment with gabapentin (50 and 100 mg/kg) significantly increased ambulatory movements, exerted anti-anxiety like effect and reduced oxidative damage in mice subjected to acute hypoxic stress. Treatment with picrotoxin (1.0 mg/kg) per se had no significant effect on behavioral and biochemical parameters of stressed mice. Treatment with muscimol (0.05 mg/kg) per se significantly increased the locomotor activity of stressed mice, exerted significant anti anxiety effect and significantly reduced the oxidative damage. Further, pretreatment with picrotoxin (1.0 mg/kg) significantly blocked whereas pretreatment with muscimol (0.05 mg/kg) significantly potentiated the neuroprotective effect of gabapentin. These results suggest that gabapentin produces its neuroprotective effect in mice subjected to acute hypoxic stress through GABA(A) receptor mechanism. PMID- 18432055 TI - Possible involvement of nitric oxide (NO) signaling pathway in the antidepressant like effect of MK-801(dizocilpine), a NMDA receptor antagonist in mouse forced swim test. AB - L-arginine-nitric oxide (NO)-cyclic guanosine monophosphate (cGMP) is an important signaling pathway involved in depression. With this information, the present study aimed to study the involvement of this signaling pathway in the antidepressant-like action of MK-801 (dizocilpine; N-methyl-d-aspartate receptor antagonist) in the mouse forced-swim test. Total immobility period was recorded in mouse forced swim test for 6 min. MK-801 (5-25 microg/kg., ip) produced a U shaped curve in reducing the immobility period. The antidepressant-like effect of MK-801 (10 microg/kg, ip) was prevented by pretreatment with L-arginine (750 mg/kg, ip) [substrate for nitric oxide synthase (NOS)]. Pretreatment of mice with 7-nitroindazole (7-NI) (25 mg/kg, ip) [a specific neuronal nitric oxide synthase inhibitor] produced potentiation of the action of subeffective dose of MK-801 (5 microg/kg, ip). In addition, treatment of mice with methylene blue (10 mg/kg, ip) [direct inhibitor of both nitric oxide synthase and soluble guanylate cyclase] potentiated the effect of MK-801 (5 microg/kg, ip) in the forced-swim test. Further, the reduction in the immobility period elicited by MK-801 (10 microg/kg, ip) was also inhibited by pretreatment with sildenafil (5 mg/kg, ip) [phosphodiesterase 5 inhibitor]. The various modulators used in the study and their combination did not produce any changes in locomotor activity per se and in combination with MK-801. MK-801 however, at higher doses (25 microg/kg, ip) produced hyperlocomotion. The results demonstrated the involvement of nitric oxide signaling pathway in the antidepressant-like effect of MK-801 in mouse forced-swim test. PMID- 18432056 TI - Beneficial effect of modified egg on serum T3, T4 and dyslipidaemia following dietary Zn-supplementation in Wistar rat. AB - A fall in serum T3 and T4 along with increase in serum cholesterol, triglycerides, LDL-c and VLDL-c and decrease in HDL-c was observed in albino Wistar rats when fed on semi-synthetic diet containing either 40 or 80mg Zn/kg diet. Zn concentrations were observed to increase with decreased concentration of Cu and Mg in their tissues. On including modified egg (Indian Patent Application No. 2264?Del?2005) in the Zn supplement diet, the levels of T3 and T4, lipid profile in serum and mineral status approached closer to control group-I. The data suggest that hypothyroidism and dyslipidaemia caused by excessive Zn in diet can be ameliorated on consuming these modified eggs due to restoration of mineral status in the body. PMID- 18432057 TI - Antidepressant activity of fosinopril, ramipril and losartan, but not of lisinopril in depressive paradigms of albino rats and mice. AB - Fosinopril, ramipril and losartan significantly decreased the duration (sec) of immobility in forced swim test and were comparable to amitriptyline. The duration of immobility were significantly decreased in fosinopril, ramipril and losartan in the tail suspension test and were comparable to amitriptyline. Only losartan significantly increased the rearing number of entries, time spent (sec) in open arm and in light area in comparison to control animals. Fosinopril and ramipril and not lisinopril showed significant antidepressant activity while losartan showed a significant antidepressant and anxiolytic activity. Present findings suggest that these drugs could be better antihypertensives in hypertensive patients with comorbidity like depression or anxiety. PMID- 18432059 TI - Surface morphology of immunocompetent cells isolated from spleen of Bufo himalayanus (Gunther). AB - Immunocompetent cells were isolated from spleen of B. himalayanus and studied surface morphology of the three different cell types--(i) plastic adherent; (ii) nylon wool adherent; and (iii) nylon wool non-adherent cells. As revealed by scanning electron microscopy, they resembled the macrophages, B and T cells, respectively. Presence of such cell types indicated that Bufo himalayanus possessed a well-organized immune system. Further work is needed to characterize the functional efficacy of these immunocompetent cells found in B. himalayanus. PMID- 18432058 TI - CNS depressive role of aqueous extract of Spinacia oleracea L. leaves in adult male albino rats. AB - Treatment with Spinacia oleracea extract (SO; 400 mg/kg body weight) decreased the locomotor activity, grip strength, increased pentobarbitone induced sleeping time and also markedly altered pentylenetetrazole induced seizure status in Holtzman strain adult male albino rats. SO increased serotonin level and decreased both norepinephrine and dopamine levels in cerebral cortex, cerebellum, caudate nucleus, midbrain and pons and medulla. Result suggests that SO exerts its CNS depressive effect in PTZ induced seizure by modulating the monoamines in different brain areas. PMID- 18432060 TI - Effect of propofol in altering pentylenetetrazol induced seizure threshold in rats. AB - The present study was undertaken to evaluate the role of propofol in altering pentylenetetrazol induced seizure threshold in rats. Total 42 Wistar rats were used to evaluate different parameters (onset of action, duration of seizure, seizure severity score and number of seizure) following propofol injection. The present results showed that there was significant reduction in the time required for onset of seizure in propofol treated groups following PTZ treatment. If treated with propofol alone (2 and 5 mg/kg), there was no significant difference as compared to controls. In seizure severity score assessment, there was no significant difference with various doses of propofol alone treated groups, but the difference was observed in propofol (2 and 5 mg/kg) treated groups following PTZ treatment. Duration of seizure also significantly increased in propofol (5 mg/kg) treated group, but at 2 mg/kg of propofol treatment, no significant difference was observed. The present results showed that propofol ameliorate seizure threshold and caused prolongation of duration of seizure. However, further study and trials are needed to confirm the present results. PMID- 18432061 TI - Thresholds of physical activities necessary for living a self-supporting life in elderly women. AB - BACKGROUND: To decide the minimal levels (thresholds) of physical activities necessary for a self-supporting life using discriminate analysis between community-dwelling elderly individuals and daycare-service-receiving elderly individuals in Japan. METHOD: A total of twenty-six elderly women, including twelve living a self-supporting life and fourteen using a daycare service, were recruited in this study. The parameters examined were physical performance, activities of daily living, exercise capacity. Discriminate analysis was used to determine the thresholds of physical activities needed to live a self-supporting life. RESULTS: Muscle masses, grip strength, lower-extremity muscle force, gait ability and balance function in elderly individuals living a self-supporting life were significant higher than those in elderly individuals using a daycare service. A threshold of physical ability over 75% classifies accurately using the methods of discriminate analysis indicate total body muscle mass, thigh muscle mass, knee extension force, 6 min walking distance (6MD), 10 m obstacle walking time, Activity of daily living (ADL) index and daily steps. CONCLUSIONS: The present study indicates that thresholds of physical activities including gait abilities, muscle force, and muscle mass are very important factors in maintaining a self-supporting life for elderly individuals. Thresholds of physical activities were more effective than standard values of physical activity for elderly individuals using a care service in meeting the goals of rehabilitation. PMID- 18432062 TI - Hippocampal BDNF and TrkB expression in young rats after status epilepticus. AB - BACKGROUND: The immature brain is more susceptible to seizures than mature brains but less vulnerable to seizure-induced neuronal loss. We studied age-related susceptibility and vulnerability to kainic acid-induced status epilepticus (KASE) in rats in terms of hippocampal expression of brain-derived neurotrophic factor (BDNF) and tyrosine kinase B receptor (TrkB). METHODS: Immunohistochemical and Western analysis were performed after kainic acid (KA)-induced status epilepticus (SE). RESULTS: KA doses required to induce SE increased from 1.5 mg/kg in 1-week old rats to 10 mg/kg at 4 weeks of older. After SE the older rats showed spontaneous seizures and hippocampal pyramidal neuronal loss-unlike rats under 4 weeks old. Hippocampal BDNF protein expression had increased fivefold in 1-week old rats and threefold in 8-week-old rats 1 day after SE, returning to baseline 2 days after SE. TrkB expression showed little effect from KASE at either age. CONCLUSIONS: These results indicated that the critical period as for vulnerability to SE was the age of 4-week-old and older in the rat. Since the response patterns of BDNF and TrkB to SE were similar between neonatal and the adult rats, our study revealed that the observed transient upregulation of BDNF did not contribute to cause epilepsy in neonatal rats. PMID- 18432063 TI - Effect of normal human erythrocytes on blood rheology in microcirculation. AB - BACKGROUND: Effects of RBCs on blood rheology were studied using a microchannel array flow analyzer (MC-FAN). METHODS: Fluidity of four types of samples prepared from the venous blood of healthy volunteers was examined in terms of passage time through the microchannel array of MC-FAN (model KH-3): (1) physiological saline RBC suspensions and plasma-RBC suspensions, each adjusted to a predetermined hematocrit value; (2) suspensions of glutaraldehyde-treated hardened RBCs; (3) fibrinogen-RBC and albumin-RBC suspensions; and (4) dextran-RBC suspensions. RESULTS: Hematocrit positively correlated with passage time. Both plasma and fibrinogen prolonged passage time significantly. Hardened RBCs completely obstructed the microchannel. The passage time of dextran-RBC suspensions was prolonged in a dextran molecular weight- and concentration-dependent manner and was dependent on the passage time of the solution alone. CONCLUSIONS: Blood rheology, as determined by MC-FAN, is affected not by RBC aggregation but hematocrit, RBC deformability, and the passage time of the solution. PMID- 18432064 TI - Enhanced expression of angiotensin II type 1 receptor in usual interstitial pneumonia. AB - BACKGROUND: Angiotensin II, a potent vasoconstrictor, has been considered to be involved in various fibrotic disorders including idiopathic interstitial pneumonias. To clarify whether this agent contributes to the development and progression of usual interstitial pneumonia, a major entity of idiopathic interstitial pneumonias, we immunohistochemically examined expression of its specific receptor, angiotensin II type 1 receptor, in human normal and diseased lung tissues. METHODS: Video-assisted thoracoscopic lung biopsy specimens obtained from patients with usual interstitial pneumonia (n=8) were sectioned and stained using single or double immunostaining techniques with specific antibodies against angiotensin II type 1 receptor and smooth muscle actin. Lung tissues of desquamative interstitial pneumonia (n=2) and normal lung tissues (n=6) were also examined for comparative analyses. RESULTS: Expression of angiotensin II type 1 receptor was limited in vascular and bronchial smooth muscle cells in normal lungs. In contrast, the receptor-positive mesenchymal cells, most of which were also positive for smooth muscle actin and arranged like a bundle, were markedly increased in association with dense collagen deposition in thickened alveolar walls of usual interstitial pneumonia. In desquamative interstitial pneumonia, the fibroproliferative change, including angiotensin II type 1 receptor-positive mesenchymal cell proliferation, was milder than that in usual interstitial pneumonia. CONCLUSIONS: These findings suggest that angiotensin II and its type 1 receptor play a profibrogenic role in idiopathic interstitial pneumonias, particularly in usual interstitial pneumonia. Furthermore, angiotensin II type 1 receptor-positive smooth muscle cells increased in diseased lung tissues may be contractile and may contribute to reduction of airspaces in usual interstitial pneumonia. PMID- 18432065 TI - Effectiveness of arsenic mitigation program in Bangladesh--relationship between arsenic concentrations in well water and urine. AB - BACKGROUND: Arsenic in drinking water remains a major public problem in Bangladesh, although arsenic mitigation programs began there a decade ago. The purpose of this study was to examine the effectiveness of this program by determining the relationship between current arsenic levels in well water and the high level of urinary arsenic excretion. METHODS: A community-based cross sectional study was conducted in the Pabna district of Bangladesh between May and July 2005. We included 174 married couples and collected their drinking water from 138 wells. The allowable limit for arsenic in drinking water is 50 microg/L in Bangladesh, while the normal level of urinary arsenic is < or =40 microg x 1.5 L(-1) x day(-1) by Dhaka Community Hospital. RESULTS: Of 348 subjects, 304 exceeded the urinary arsenic level of 40 microg x 1.5 L(-1) x day(-1). Of all wells, 44.2% had arsenic levels >50 microg/L. Multiple-adjusted odds ratios of urinary arsenic level >40 microg x 1.5 L(-1) x day(-1) were 8.90 (95% CI: 3.31 23.93) for the arsenic level in well water of 11-50 microg/L, and 53.07 (11.91 236.46) for that of 51-332 microg/L, compared with < or =10 microg/L. When the Bangladeshi standard arsenic level in drinking water of 50 microg/L was used, the sensitivity in detecting subjects with a urinary arsenic level >40 microg x 1.5 L(-1) x day(-1) was 50%, although when the World Health Organization (WHO) guideline value of 10 microg/L was used, it was 76.3%. CONCLUSIONS: Green marked wells, which the Bangladesh government regards as safe, are not always safe. The mitigation programs should use the WHO guideline arsenic level to determine the safety of well water for drinking. PMID- 18432066 TI - A Schloffer tumor treated 27 years after appendectomy. AB - Most complications of appendectomy occur in the early postoperative period. A 51 year-old woman presented to our hospital with abdominal fullness and a painful mass in the right lower abdomen. The painful mass was related to a scar from an appendectomy performed 27 years previously; some earlier episodes of pain had occurred after the operation. A tender mass 7 cm in diameter was palpable, extending from the appendectomy scar toward the umbilical region. Findings of abdominal ultrasonography and computed tomography were consistent with an abdominal wall abscess, which was excised. At operation, a thick-walled abscess was seen to extend from the appendectomy scar toward the umbilicus, and from within abdominal wall muscles to the preperitoneal fat; it did not communicate with any intra-abdominal organ. Bacteroides fragilis was isolated from the abscess contents. Pathologic examination revealed diffuse infiltration by inflammatory cells including neutrophils and plasma cells, with inflammation spreading into surrounding muscles. Our patient's findings indicate that the abdominal wall abscess (so-called Schloffer tumor) resulted from persistent inflammation following the appendectomy performed 27 years previously. Since treatment of Schloffer tumor is not difficult once the diagnosis is made, physicians and surgeons should be aware of Schloffer tumor as a possible late complication of appendectomy. PMID- 18432067 TI - Hemorrhagic small intestine cancer with solitary pulmonary metastasis initially presented as suspected primary lung cancer: an autopsy report. AB - Cancer of the small intestine presenting with a solitary pulmonary metastasis is rare. Diagnosis and treatment of hemorrhagic small intestinal disease is clinically problematic due to its anatomic aspect, especially after multiple laparotomies. The case that we present here was a 79-year-old man who was initially diagnosed with suspected T2N2M0 lung cancer. After non-diagnostic results on two bronchoscopic biopsies and computed tomography-guided needle biopsy, he was admitted for thoracoscopic biopsy and possible curative operation. The patient had a history of multiple laparotomies for gastric ulcer and had no abdominal symptoms. A fecal occult blood test was positive; this was thought to be because of persistent bloody sputum. During the preoperative evaluation period, massive intestinal hemorrhage occurred. Intestinal tumor was identified by double-balloon enteroscopy and emergency laparotomy was performed to control the bleeding. The histopathological diagnosis was metastatic adenocarcinoma. However, intestinal bleeding started again. His systemic status deteriorated progressively, resulting in death. Autopsy revealed a large polypoid tumor with hemorrhagic necrosis in the jejunum that was histologically and immunohistochemically diagnosed as primary poorly differentiated adenocarcinoma in the small intestine. Multiple small submucosal tumors with central ulceration were confirmed as intramural metastases. A lung mass in the right lower lobe was diagnosed as a metastatic lesion. In the diagnosis and treatment of the disease, we faced several clinically difficult problems. We here describe in detail the clinical course and the diagnostic and therapeutic difficulties of this rare case, with some references to the literature. PMID- 18432068 TI - [Are we able to appreciate experience of our teachers?]. PMID- 18432069 TI - [Variations of primary spontaneous pneumothorax management]. AB - OBJECTIVE: The aim of our study was to determine the current practice and variation among pulmonologists, general and thoracic surgeons treating primary spontaneous pneumothorax (PSP) in the Czech Republic. METHODS: A pneumothorax questionnaire was mailed to 153 chiefs of departments of general surgery, pneumology and to thoracic surgeons (55 pulmonologists, 75 general surgeons, 23 thoracic surgeons). Mail contained the survey questionnaire and cover letter requesting the recipients' participation and explaining the purpose of the research. National survey questionnaire included questions on PSP management, chest tube attachment preferences and its removal management RESULTS: Ninety seven respondents (63%) answered the survey. This included 28 pulmonologists (28/55, 52%), 48 general surgeons (49/75, 64%) and 20 thoracic surgeons (20/23, 87%). Sixty nine percent of respondents treat first PSP with chest tube, 6% by aspiration. Seventy four percent of respondents insert chest tube in the second intercostal space midclavicular line and 22% in the fourth intercostal space midaxillary line. Nearly 71% of pulmonologists use < 18F chest tube, 85% of thoracic surgeons insert a 20-24F chest tube. Active suction use 56% of respondents. Seventy four percent of physicians clamp chest tube prior its removal. Thirty nine percent of respondents pull out a chest tube at the end of inspirium, and 33% at the end of expirium. CONCLUSIONS: Marked practice variations exist among physicians treating spontaneous pneumothorax. Variations exist not only between pulmonologists and surgeons, but also in the group of surgeons itself. PMID- 18432070 TI - [Colorectal carcinoma as a cause of acute abdominal illness]. AB - Colorectal carcinoma (KCR) is the commonest malignancy in male patients and the second commonest in female patients in the Czech Republic. During 1990-2006, 1162 patients with colorectal carcinomas were operated in the FTNsP (Faculty Thomayer Hospital) Surgical Clinic. 212 patients aged between 39 to 94 y.o.a.(the median of 70 y.o.a) were managed urgently for acute abdominal illness. In this group of urgently managed patients, the mortality rate was 17% and the morbidity rate was 38%. Future prospects of any colorectal carcinoma patient with acute abdominal illness depend on the procedure's radicality, which is limited by the overall patient's condition. The preoperative care aims to improve the patient's overall condition to such a degree, to allow for surgery fulfilling principles of oncosurgical radicality. According to this report's data, such urgent surgical procedures do not result in increased mortality or morbidity rates, compared to these in planned procedures, and show the best results in this patient group. PMID- 18432071 TI - [Diagnostics of intestinal ischemia. Influence of surgery on plasma levels of I FABP as the marker of enterocyte injury]. AB - BACKGROUND: Intestinal fatty acid binding protein (I-FABP) deriving from the cytoplasm of the intestinal epithelial cells is a new biochemical marker with potential to indicate intestinal mucosal injury early in the phase of enterocyte damage. I-FABP is thought to be promising marker for detection of the systemic inflammatory response syndrome (SIRS) and sepsis before its onset. AIMS OF THE STUDY: The aim of the study was to consider the influence of surgery on I-FABP plasma levels. METHODS: Fifty-six consecutive patients undergoing elective surgery were prospectively studied. Only patients with major surgical procedure were involved in the study. Patients were divided into three groups: group A (n=19) patients with open abdominal surgery, group B (n=19) patients with laparoscopic abdominal surgery and group C (n=15) patients with thoracic surgery. From every patient in the study six samples of venous blood were taken in these intervals: preoperatively, immediately postoperatively and 4, 12, 24 and 48 hours after the surgery. Plasma samples were analysed for I-FABP levels by enzyme linked immunosorbent assay. RESULTS: There were no differences in the mean plasma I-FABP levels within the groups (ANOVA, p > 0.05), except significant decrease of the I-FABP level 48 hours after the surgery in contrast to preoperative plasma concentration in group B (82.60 pg/ml vs. 229.00 pg/ml; p = 0.025) and group C (88.99 pg/ml vs. 194.96 pg/ml; p = 0.0347). There were no differences in the mean I-FABP plasma concentrations among the groups preoperatively, immediately postoperatively and 4, 12. 24 and 48 hours after the surgery (ANOVA, p > 0.05). No differences were found in the I-FABP plasma levels related to preoperative laxative drug use, duration of surgery, small bowel resection and SIRS in postoperative period (t-test, p > 0.05). CONCLUSIONS: There were no differences in the mean plasma I-FABP levels preoperatively and in the postoperative period after the major surgery. Because surgery alone probably has no influence on I FABP plasma levels we can say that the concept of l-FABP as the early marker of the intestinal mucosal injury is also viable in postoperative period. PMID- 18432072 TI - [Postoperative nutritional support in liver surgery. Effects of specialized parenteral nutrition enriched with branched-chain amino acids following liver resections for colorectal carcinoma metastases]. AB - INTRODUCTION: Perioperative nutritional support in liver surgery remains specific regarding the role of the liver in the metabolism regulation. The loss of functional liver tissue following resection procedures may result in significant metabolic disorders, particularly in patients with preoperative liver impairment in chronic conditions. Perioperative nutritional support is indicated in patients suffering from malnutrition, chronic liver disorders and also following liver resections with limiting residual parenchyma. AIM OF THE STUDY: The retrospective study assessed the effect of complete parenteral nutrition enriched with branched chain amino acids on liver function and the rate of postoperative complications following liver resections (of at least two segments) for colorectal carcinoma metastases. SUBJECTS AND METHODOLOGY: 142 subjects following liver resections for colorectal carcinoma metastases were included in the study. In 42 subjects with anatomical resections of at least two segments, complete parenteral nutrition enriched with branched chain amino acids (3.8 g) was administered postoperatively from Day 1. The parenteral nutrition was maintained for 5 to 7 days, from Day 3, additional oral diet regime was indicated. In 100 subjects with a single segment resections or wedge resections, a combination of glucose 10% and crystalloid solutions was administered from Day 1, and early oral diet was added from Day2. Liver tests were performed during the first postoperative week and protein electrophoresis 14 days postoperatively. RESULTS: During the first postoperative week, the liver test values reached normal values in patients with specialized complete parenteral nutrition. In subjects, where the specialized parenteral nutrition was not administered, increased alcaline phosphatase levels and gamma glutamyl transpherase levels were recorded through the postoperative Day 7. No significant differences were detected in protein electrophoresis values and in rates of postoperative complications (14% in both patient groups). Liver test values in specialized nutritional support patients were negatively affected by the extent of their liver resections and occurrence of additional extra hepatic disorders. Administration of complete parenteral nutrition enriched with branched chain amino acids proved a positive effect on the liver test values recovery. CONCLUSION: Postoperative parenteral nutrition with solutions enriched with branched-chain amino acids in major liver resections (at least 2 segments) has a positive impact on the liver test values recovery during the early postoperative period. PMID- 18432073 TI - [Implementation of POSSUM scoring system in assessing morbidity after laparoscopic colorectal surgery]. AB - AIM: The aim of the study was objective morbidity comparison of the open and laparoscopic colorectal surgery using the recalibrated POSSUM scoring system, taking into account potential "case mix" of samples. MATERIAL AND METHODS: In total 510 patients participated in the study, undergoing an elective colorectal surgery between 1st January 2001 and 31st January 2004. The samples of patients were operated either by an open or laparoscopic technique were compared. The study was retrospective-prospective, non-randomized. The basic data of the patient, data of the POSSUM scoring system (physiological score, operative score), data describing the surgery, post-operative course and complications were monitored. The individual risk of morbidity was evaluated by the modification (recalibration) of the original model of the POSSUM for the sample of patients operated by the open technique. Recalibration was carried out by the method of logistic regression analysis and tested. The real morbidity in relation to the individual risk of patients in both samples was afterwards statistically analyzed. P value < 0.05 was considered statistically significant. RESULTS: The complete data were obtained from 274 patients operated by the open technique and 231 patients operated laparoscopically. Although it was not a randomized study, both samples were comparable in basic characteristics (sex, age, BMI, ASA classification, surgery localization, type of procedure). In the group operated by the open technique, the patients with a malignant disease were represented more often and there was also a higher portion of palliative surgeries. The samples did not differ significantly in operative time, blood loss or frequency of per-operative complications. As far as post-operative complications are concerned, the post-operative bleeding into the abdominal cavity was recorded more often with the group operated by the laparoscopic technique, while the group operated by the open technique exhibited a higher occurrence of cardiac, circulation and neurological complications. The mortality in the group operated laparoscopically did not significantly differ from the group operated by the open technique (3.5% resp. 3.3%), the morbidity was significantly lower in the laparoscopic group (27% resp. 39%). The original POSSUM model recalibrated by the logistic regression analysis confirmed its compactness with the starting data and coefficient significance. The statistical analysis of morbidity related to the individual risk groups derived by the modificated (recalibrated) POSSUM proved significantly lower morbidity with the patients operated laparoscopically (p < 0.05). CONCLUSION: The elective laparoscopic colorectal surgery is associated with lower post-operative morbidity than the open approach, including high-risk patients. PMID- 18432074 TI - [K-ras mutation as a prognostic factor in colorectal carcinoma]. AB - INTRODUCTION: Colorectal carcinoma presents a serious problem in the Czech Republic: its incidence is on the increase and--according to some statistics takes first place among developed countries worldwide. Therefore, it is advised to incorporate examinational and the rapeutic algorithms with new modalities that will lead to early diagnostics or to a change in existing therapeutic procedures. CHARACTERIZATION OF K-RAS MUTATION: K-ras mutation belongs to the family of protooncogenes where a gene not having undergone mutation expresses proteins that regulate mitosis. Mutation cancels the regulatory function of these proteins, thus leading to the develop ment of tumors, especially carcinoma of the lungs, pancreas, and colorectum. PROJECT OBJECTIVE: The main objective of the project is to prove K-ras mutation in tumors of the colorectum: to detect tumor cells with K ras mutation in peripheral blood; to detect K-ras mutation in liver metastases: and to verify the hypothesis claiming that tumors with K-ras mutation have a worse prognosis and often lead to disemination, mainly to the liver. METHODOLOGY AND COLLECTION OF DATA: The whole project is tied to an IGA grant and runs according to the strict rules of the protocol applied at the Surgical Clinic of the Pardubice Hospital, with its diagnostic part--PCR analysis being completed at the Biochemical Diagnostic Institute (UKBD) of the Teaching Hospital in Hradec Kralove. RESULTS: The project has been running since June, 2004 to December 2006. 76 patients meeting defined parameters have been included in the file to date. K ras mutation has been detected in the tumor tissue of 25 patients (33%). K-ras mutation hasn't been detected in the blood. DISCUSSION: Genetically analysis of a specific tumor has not yet become a standard part of the examinational and therapeutic algorithm. If an assumption of a worse course of illness and metastasizing--especially to the liver has been proven, the examination of Kras mutation in patients suffering from colorectal carcinoma should lead to the adjustment of their treatment and postoperative dispensarization, or the administration of chemotherapy and radiotherapy at stages when these modalities are not normally applied. PMID- 18432075 TI - [Periampullary gangliocytic paraganglioma]. AB - We present the case of very rare periampullary gangliocytic paraganglioma in which we preferred proximal duodenopancreatectomy to local resection of the ampula of Vater because of not clear preoperative pathohistology and biologic behavior of the tumor. Although the definitive examination of the specimen proved its benign character we consider our procedure reasonable. PMID- 18432077 TI - [Polytraumas in injured patients over 65 years of age]. AB - INTRODUCTION: Geriatric trauma is most commonly defined as a trauma in patients of 65 years of age and over. These patients represent a specific problem due to their vulnerability, limited physiological response to the traumatic stress and high frequency of associated disorders, complicating the treatment. The aim of this report is to assess polytrauma patients of 65 years of age and older, compared to a group of younger patients. PATIENTS, METHODOLOGY AND RESULTS: 198 polytrauma patients, hospitalized from 2005 to 2006 were included in the retrospective study. Out of the total, 27 subjects (13.6%) were 65+ years old. Upon their admission, the difference in APACHE II between the groups was highly statistically significant (19-27.2, p = 0.0001). The ISS (the mean value) score was higher in the elderly patients (29-38). Traffic injuries, including pedestrian-vehicle collisions (61-73%), were the commonest mechanism of injuries in the both groups. There was a significant difference in the rates of craniocerebral injuries (80-96%, p = 0.041). During the early posttraumatic period, 99 (57%) urgent surgical procedures were performed in younger patients, compared to 11 (79%) procedures in the elderly. The difference in death rates was statistically significant (25-44%, p = 0.031). CONCLUSION: Polytraumas in the elderly (65 years of age and over) injured is characterized by higher severity scores and higher rates of craniocerebral injuries. Traffic injury was the commonest mechanism of injury. The death rate was higher in the elderly patients. PMID- 18432076 TI - [New options for management of posttraumatic articular cartilage defects]. AB - Articular cartilage trauma, in particular due to its poor healing potential remains a complicated problem in both the adult and paediatric traumatology and orthopedics. In older patients, total endoprosthesis of the joint is a method of choice, however, in younger patients, the situation remains more complicated. In case of osteochondral lesions (arthrosis, chondral fractures. osteochoodrosis dissecns) the ideal management should result in complete recovery of the hyaline cartilage on the traumatized joint surface. Contemporary medicine uses some therapeutic procedures resulting in partial recovery of the articular cartilage structure at the lesion site and several techniques of excisionining the articular surface's injured part and of transplantations of biological grafts. Regarding the above first approach, abrasive methods (micro fractures, small drill holes), which are expected to result in recovery of the articular cartilage through progenitor cells that migrate from the bone marrow to the defect site following subchondral fracturing. In case the injury is managed early, the osteochondral fragment may be fixed and the articular congruence be recovered. Mosaicoplasty using osteochondral auto grafts or other autologous grafts, or more recently using transplantations of autologous chond rocytes, which seem to have a major potential in the hyaline cartilage healing process. However, methodology of the transplant retention at the defect site remains a problem. Furthermore, the use of mesenchymal stem cells, so far in the experimental phase, appears prospective. Pivotal articular cartilage treatment research activities have progressed to a level of searching for a suitable scaffold of perfect qualities. This is the task for cooperation with bioengineering. requiring provision of the most exact differentiation protocol for hyline cartilage producing mesenchymal stem cells (MSCs). PMID- 18432078 TI - [Dual kidney transplantation]. AB - Despite the increasing demand for organs for transplantation, the number of cadaveric donors remains stable and waiting time for transplantation is gradually getting longer. In addition to the options of using kidneys from living donors and those of non heart-beating donors (NHBD), an alternative approach is transplantation of both kidneys from adult marginal donors who would otherwise be considered unsuitable for single-kidney donation. Dual kidney transplantation involves the use of both kidneys from a marginal donor for a single recipient without the recipient having to cope with the drawbacks of a limited number of functioning nephrons. Normally. these kidneys would be excluded from the transplantation program and remain unused. The submitted presentation provides information on donor and recipient selection criteria and describes the course of the first dual kidney transplantation in an adult recipient in the Czech Republic. PMID- 18432079 TI - What are the risk factors for nausea and vomiting after neurosurgery? A systematic review. AB - In their practice of post-operative neurosurgical care, nurses frequently encounter the problem of post-operative nausea and vomiting (PONV). Knowledge of risk factors for PONV can help nurses to identify groups of patients who may encounter PONV and develop related therapeutic guidelines. The purpose of this systematic review was to identify and summarize risk factor research for PONV after neurosurgical procedures. Of 272 papers identified through a systematic search, 13 studies met the criteria for this review. These studies varied considerably in their outcome measurements, risk factors studied, and target populations. The timeframe of observation (for prospective studies) or chart review (for retrospective studies) ranged from one hour after surgery to the entire length of the hospital stay. For these reasons, it was difficult to compare results among the studies. Overall, the methodological quality of the studies was fair, with few studies controlling for confounders and many with limited explanations of how the risk factors and/or outcomes were measured. Despite these limitations, many authors looked at unique risk factors for neurosurgical patients such as location of surgery (i.e., infratentorial versus supratentorial), awake versus general anesthesia, and use of fat grafting for cerebral spinal fluid leak in transsphenoidal procedures. The findings may challenge traditional thinking about risk factors for PONV and guide future studies. PMID- 18432080 TI - Living with oculopharyngeal muscular dystrophy: a phenomenological study. AB - Oculopharyngeal muscular dystrophy (OPMD) is an autosomal dominant form of late onset muscular dystrophy. Ptosis (droopy eyelids) and dysphagia (difficulty swallowing) are the most common presenting symptoms. The purpose of this phenomenological study was to describe the experience of living with OPMD. Purposeful sampling was used to recruit individuals with genetically confirmed OPMD who displayed ptosis and dysphagia, were 40 years or older, English speaking, and were willing to consent to the tape-recording of the interviews. An unstructured interview format was used to solicit the participants' perspectives of living with droopy eyelids, difficulty swallowing, and a genetic disorder. The interviews were audiotaped and transcribed verbatim. Colaizzi's Method was used to analyze the data, which identified five comprehensive themes. The themes that emerged describing the experience of living with OPMD were "Adjusting to Change", "Managing Misconceptions", "Seeking Normality", "Facing the Future", and "Informing Children". The information derived from this study will assist nurses to identify the burdens of living with OPMD and to intervene appropriately early in the course of the disease. PMID- 18432081 TI - Did you know? Salt and sodium. PMID- 18432082 TI - ONS can help you stay current in the many subspecialties of oncology nursing. PMID- 18432083 TI - From then to now. An update on blood and marrow transplantation. PMID- 18432084 TI - What changes have you seen in stem cell transplantation in the past 10 years? PMID- 18432085 TI - How do you support caregivers of patients receiving outpatient transplantation? PMID- 18432086 TI - Nurses discuss working with challenging patients. PMID- 18432087 TI - Recognize hepatic sinusoidal obstruction syndrome in patients with cancer. PMID- 18432088 TI - Elasticity of cells may help researchers to identify cancer. PMID- 18432089 TI - Your thoughts and comments are a valued decision-making resource. PMID- 18432090 TI - Distribution of tissue enzymes in three species of pinnipeds. AB - In domestic animal medicine, changes in serum enzyme levels are routinely used as diagnostic tools to detect liver disease. Hepatic disease occurs in pinnipeds, but limited data are available on the tissue distribution of serum enzymes in marine mammals. The objectives of this study were to determine the tissue distribution of seven serum enzymes in three pinniped species. Enzymes evaluated were alanine aminotransferase (ALT), aspartate aminotransferase (AST), sorbitol dehydrogenase (SDH), lactate dehydrogenase (LDH), creatine kinase (CK), alkaline phosphatase (ALP), and gamma-glutamyl transferase (GGT) in tissues from California sea lions (Zalophus californianus) (n = 5), harbor seals (Phoca vitulina) (n = 5), and northern elephant seals (Mirounga angustirostris) (n = 5) that stranded and then died at a rehabilitation center. Samples were evaluated in duplicate from liver, skeletal muscle, cardiac muscle, kidney, adrenal, spleen, pancreas, lung, lymph node, and intestine. Patterns of tissue enzyme distribution were similar in all species, with SDH activity highest in liver and kidney, CK activity highest in skeletal and cardiac muscle, ALP activity highest in adrenal, and GGT activity highest in the kidney. Aspartate aminotransferase and LDH activities were less specific, with high activity in multiple tissues. Tissue ALT activity was high in the liver of all species, but was also high in cardiac muscle (California sea lions), skeletal muscle (harbor seals), and kidney (elephant seals). These results suggest that concurrent analysis of SDH, ALT, and CK would provide high specificity and sensitivity for the detection of hepatic lesions, and allow differentiation of liver from skeletal muscle lesions in pinniped species. PMID- 18432091 TI - Blood-gas and acid-base parameters in nontranquilized Arabian oryx (Oryx leucoryx) in the United Arab Emirates. AB - Arterial and venous blood-gas and acid-base values were established from a herd (n = 19; 14 male, 5 female) of semi-free-ranging Arabian oryx (Oryx leucoryx) in the United Arab Emirates. The animals were restrained with the use of a modified raceway incorporating a commercially available handling crate. Statistically significant differences were found between arterial and venous values for PO2 (p < 0.001), PCO2 (p = 0.0141), SO2 (p < 0.001), pH (p = 0.0494), and glucose (p < 0.0001). The results are similar to those reported for the same species under field anesthetic conditions, and to those reported from other species of wild bovidae, both tranquilized and nontranquilized, established under similar methods of restraint. In addition, Bland and Altman plots suggest adequate levels of clinical agreement between venous and arterial pH but not between arterial and venous PCO2. PMID- 18432092 TI - Trapping and handling of North American river otters (Lontra canadensis) in a managed marsh. AB - North American river otters (Lontra canadensis) were trapped in a managed marsh in northern California between July and November of 1998. Five trap types using several set configurations were utilized in this study. Otters were successfully captured with minimal injury and a relatively high trap rate (1 capture per 48 trap nights), with the use of No. 1.5 and No. 1.75 double-coil spring traps, and No. 11 double long spring traps on short chains in blind land sets, or in bank sets on one-way cables leading to land-buried stakes. Only 3 of 14 captures incurred more than minimal injuries, and all 3 of these were exacerbated by complications from traps attached to long chains. Otters were captured at any time of day, although activity appeared lowered between 0900 and 1900 hours. Otters were anesthetized for handling. Monitoring results and anesthetic complications are reported for 14 captures. Ketamine (15 mg/kg) and midazolam (0.5 mg/kg) provided a wide margin of safety, rapid induction at low injection volume, good anesthetic quality, handling time of about 30 min, and few complications. White blood cell counts were taken and appeared high relative to reported values in other studies. This trend is likely attributable to blood draws immediately after the animal had been held in a trap for several hours, in contrast to most other studies, in which blood was drawn days to weeks after being held in captivity. PMID- 18432093 TI - Hematology, blood chemistry, and bacteriology of the free-ranging Mexican beaded lizard (Heloderma horridum). AB - Blood samples from 16 adult, wild Mexican beaded lizards were obtained for complete blood count and serum biochemical analysis. In addition, both oral and cloacal swab samples were collected and submitted for bacteriologic culture. This is the first report of hematologic and bacterial data from free-living beaded lizards. This information will serve as baseline reference values for future health assessment studies in conservation, captive breeding, and research programs. PMID- 18432095 TI - Gross and histologic evidence of sharp and blunt trauma in North Atlantic right whales (Eubalaena glacialis) killed by vessels. AB - Vessel-whale collision events represented the ultimate cause of death for 21 (52.5%) of the 40 North Atlantic right whales (Eubalaena glacialis) necropsied between 1970 and December 2006. Injuries seen in vessel-struck whales fall into two distinct categories: 1) sharp trauma, often resulting from contact with the propeller, and 2) blunt trauma, presumably resulting from contact with a vessel's hull. This study analyzes four trauma cases that resulted from vessel-whale collisions, which together provide a framework for a more critical understanding of lethal blunt and sharp trauma resulting from vessel collisions with right whales. In case no. 1, contact with a propeller resulted in three deep lacerations. The animal survived acute trauma only to succumb nearly 14 years later when the lesions reopened and became infected. In case no. 2, anecdotal reports linked the laceration of large arteries of the peduncle and histologic evidence of perimortem trauma at a bone fracture site to vessel-whale collision trauma. Case no. 3 had a laceration of the oral rete and a fracture of the rostrum. Both of the areas displayed histologic evidence of perimortem blunt trauma. Finally, in case no. 4, an antemortem mandibular fracture, two additional skull fractures, and widespread hemorrhage were consistent with severe blunt trauma. Evidence from each case, including the timing of trauma relative to the time of death and identifying characteristics of both trauma types, are presented. Before this study, no detailed comparative analysis of trauma pathology that resulted from lethal interactions between vessels and right whales had been conducted. This study demonstrates the importance of detailed gross and histologic examination in determining the significance and timing of traumatic events. This work represents a new paradigm for the differential diagnosis of lethal sharp and blunt trauma in right whales hit by ships and will enhance the present understanding of the impact of anthropogenic mortality on this critically endangered species. PMID- 18432094 TI - Serosurvey of pathogens in domestic dogs on the border of Noel Kempff Mercado National Park, Bolivia. AB - The threat of disease transmission from domestic animals to wildlife has become recognized as an increasing concern within the wildlife community in recent years. Domestic dogs pose a significant risk as reservoirs for infectious diseases, especially for wild canids. As part of a multifaceted ecologic study of maned wolves and other canids in the large, remote Noel Kempff Mercado National Park (NKMNP) in northeastern Bolivia, 40 domestic dogs in two villages and at two smaller settlements bordering the national park were sampled for exposure to canine diseases. High levels of exposure were found to canine distemper virus and canine parvovirus, both of which are known to cause mortality in maned wolves and other carnivores. Moderate to high levels of exposure were found to rabies virus, Ehrlichia canis, and Toxoplasma gondii, as well as significant levels of infection with Dirofilaria immitis. This study reports evidence of exposure to several diseases in the domestic dogs bordering the park. Contact between wild carnivores and dogs has been documented in the sampled villages, therefore dogs likely pose a substantial risk to the carnivores within and near NKMNP. Further measures should be undertaken to decrease the risk of spillover infection from domestic animals into the wild species of this region. PMID- 18432096 TI - Salmonella surveillance in a herd of asymptomatic captive black rhinoceros (Diceros bicornis) using fecal culture and PCR. AB - Feces were collected from captive black rhinoceros (Diceros bicornis minor) housed at Disney's Animal Kingdom to examine the frequency of Salmonella spp. shedding in asymptomatic animals using enrichment culture and broth culture- polymerase-chain-reaction (PCR) for detection. Three samples per animal were collected during the first week of each month between February 2001 and December 2003. During the study period, six different individual animals from one herd participated in the study, including two growing calves. A total of 550 cultures, using duplicate samples at two different laboratories, and 464 PCR tests were performed. When culture and PCR results were compared by the same laboratory, similar herd prevalence was found (2.4% positive cultures compared with 2.6% positive PCR tests). However, even though tests were performed on replicate samples, not every sample that was positive by culture was positive by PCR and vice versa. These results suggest that using multiple diagnostic methods and increasing the number of samples submitted may increase the likelihood of finding an asymptomatic Salmonella shedder. Although all of the rhinos shared the same environment throughout the study period, only four out of the six animals tested shed Salmonella spp. even though a minimum of 37 fecal samples were taken from each of the negative animals. Although this study followed a small number of rhinoceros, it suggests that asymptomatic shedding probably occurs more frequently in captive black rhinoceros than was previously believed. The prevalence appears to be similar to that reported for domestic ungulates. PMID- 18432097 TI - Hip dysplasia in koalas (Phascolarctos cinereus) at the San Diego Zoo. AB - A retrospective/prospective radiographic study documented 55 cases of moderate to severe hip dysplasia, with varying degrees of shallowing of the acetabulum, flattening or loss of the femoral head, widening or loss of the femoral neck, and femoral diaphyseal abnormalities in northern koalas (Phascolarctos cinereus) in the San Diego Zoo (San Diego, California, USA) collection. For the retrospective study, historic radiographs were examined when availble. For the prospective study, three standard views (ventrodorsal extended leg, ventrodorsal frog leg, and lateral extended leg) were used. A scoring system was developed using four areas (acetabulum, femoral head, femoral neck, and femur) and ranges of 0 to 5 (0 = not affected to 5 = severely affected) for each area, creating a total score out of 40. Scores were graded as follows: 0-2 = normal dysplasia; 3-6 = mild dysplasia; 10-19 = moderate dysplasia; and 20-40 = severe dysplasia. Thirty koalas were graded as severe, 25 koalas as moderate, and 38 koalas as excellent or mild. Affected koalas may or may not demonstrate gait abnormalities. Mild to severe degenerative joint disease may develop and symptoms may be alleviated with glucosamine/chondroitin sulfate and nonsteroidal anti-inflammatory drugs. The etiology of hip dysplasia in koalas is not currently understood. PMID- 18432098 TI - Irregular tooth wear and longevity in captive wild ruminants: a pilot survey of necropsy reports. AB - Tooth wear is often suggested as an important factor limiting the life span of free-ranging wildlife. Given the frequent occurrence of poor dental health in captive animals reported in the literature, one would expect tooth health to be a limiting factor in captivity as well. Additionally, it could be assumed that brachydont (browsing) animals are more susceptible to dental health problems than are hypsodont (grazing) animals, given current indications for systematic increased tooth wear in some browsing species. A pilot survey of necropsy reports of adult captive wild ruminants (n = 294, 12 species) in one facility was performed in order to test these hypotheses and to calculate the incidence of irregular tooth wear. The overall incidence of irregular tooth wear was 20%, with a very high proportion of reports that did not mention the teeth at all. In contrast to this study's hypotheses, animals with irregular tooth wear were older than animals that died from other causes, indicating that reaching above-average age was a prerequisite for the development of reported abnormalities in this data set. A grazing species (blackbuck, Antilope cervicapra) was most affected, whereas two browsing species were not affected. Affected species had been regularly fed on sandy soil, whereas browsers had received feeds from racks, indicating that husbandry practices are most important for dental health. There was a high proportion of reported serous fat atrophy in animals with irregular tooth wear, indicating the clinical relevance of the problem. On average, adult individuals of the species investigated reached 41% of the maximum reported life span. Although this number appears low, the lack of comparative data from other facilities does not allow for conclusions on the adequacy of the husbandry practices used. PMID- 18432099 TI - Biomedical evaluation of free-ranging red ruffed lemurs (Varecia rubra) within the Masoala National Park, Madagascar. AB - Complete health assessments were performed on 22 adult red ruffed lemurs (Varecia rubra), comprising nine males and 13 females, found within the Masoala National Park in northeast Madagascar. Each animal was anesthetized using tiletamine and zolazepam and underwent a thorough physical examination, including measurement of its weight and vital signs; blood collection for hematology, plasma total protein concentration, serum chemistries, fat-soluble vitamins, trace minerals, assessment of iron metabolism, toxoplasmosis serology, viral serologies, and examination for hemoparasites; fecal collection for bacterial culture and parasite examination; and collection of a representative number of any ectoparasites. Comparison of blood values with those of captive lemurs demonstrated a number of significant differences thought to be associated with physiologic state (e.g., reproductive stage and stress), hydration, and diet. There was no evidence of serious infectious diseases, and hemoparasites were not detected. The enteric flora appeared unremarkable; however, results may have been skewed toward more cold-tolerant bacteria. The fecal parasite burden was low. Lemurostrongylus spp. was identified in two of the lemurs, and there were moderate numbers of Laelapidae mites present on approximately one third of the lemurs. This study demonstrated the substantial amount of data that can be collected from free-ranging populations, considered invaluable in the management of captive populations, in reducing the incidence of captivity-related diseases, and in the risk assessment associated with reintroduction programs. PMID- 18432100 TI - Juvenile mortality in captive lesser kudu (Tragelaphus imberbis) at Basle Zoo and its relation to nutrition and husbandry. AB - Since 1956, when the Basle Zoo (Switzerland) initiated the breeding of lesser kudu (Tragelaphus imberbis), 43% of the lesser kudu juveniles died before reaching an age of 6 mo. In this study, the objective was to obtain the pathological findings, nutritional history, and family tree information in order to evaluate the influence of husbandry on juvenile mortality in these animals. The main cause of death was white muscle disease (WMD), diagnosed in 14 cases (26%) of the deceased juveniles. Although enclosure size had remained constant and animal accessibility to the public was constantly high, both herd size and juvenile mortality had increased from 1956-2004. The diet consumed by the whole group in 2004 had deficient levels of vitamin E and selenium. The increasing linear trend of the mortality rate since the 1960s was significant, and there was a significant correlation between herd size and overall juvenile mortality. In contrast, there was no correlation between herd size and the occurrence of juvenile mortality associated specifically with WMD. Other investigated factors (sex, inbreeding, and season) had no significant effect on overall mortality up to 6 mo of age or on mortality associated with WMD. These results characterize both a dietary and a husbandry problem, and are supported by a lack of similar juvenile mortality in another facility where the diet was supplemented with vitamin E, animal numbers were kept low, and the enclosure structure offered more retreat options for the animals. PMID- 18432101 TI - Lungworm infection in three loggerhead sea turtles, Caretta caretta. AB - Although lungworms are known to infect many mammalian species and a few are known to infect snakes, lizards, and birds, previously none were known to infect chelonians. This study documents the first three known cases of lungworms in loggerhead sea turtles, Caretta caretta. It is unlikely that the lungworms were the primary cause of illness in any of the cases, and they may be only contributory or even incidental. Changes observed in the two cases that died included tracheal and bronchial epithelial hyperplasia and goblet cell hyperplasia. Lesions caused directly by the parasites seem to be restricted to the upper respiratory tree (trachea and main bronchi), but changes in the lungs themselves may be caused by the debris produced by the worms. Although neither case was successfully treated for the lungworms prior to death, it would appear that oxfendazole may be an effective treatment and has been used in the third case that is still undergoing rehabilitation. PMID- 18432102 TI - Repair of horn and frontal bone avulsion in a forest buffalo (Syncerus caffer nanus) with a polymethylmethacrylate dressing. AB - A 2-yr-old female forest buffalo (Syncerus caffer nanus) presented with acute trauma to the right horn following a fight with a conspecific resulting in fracture and avulsion of the frontal bone, cornual process, horn, and juxtaposed skin. The animal was sedated for wound care and debridement followed by placement of an antibacterial-impregnated polymethylmethacrylate (PMMA) dressing. The dressing was noted to be intact on examination at day 7 post-injury, but had become dislodged and had to be replaced at day 11 postinjury. The replacement dressing ultimately fell off 68 days post-injury, revealing a healed circular scar. The use of PMMA in this case provided optimal wound care. In the 6 yr since the injury, the animal has grown a nearly normal-appearing horn, suggesting that the noncornual skin and associated skull structures may retain the ability to differentiate into a near-normal appearing horn because the entire horn apparatus in this case was avulsed. PMID- 18432103 TI - Lacrimal cystadenoma in a Chinese box turtle (Cuora flavomarginata). AB - An adult male Chinese box turtle (Cuora flavomarginata) presented to the Avian and Exotic Pet Service of the Animal Medical Center for periorbital swelling of the right eye. The swelling had failed to respond to nutritional supplementation and parenteral administration of vitamin A. What had initially presented as periorbital swelling developed into a growth ventral to the globe that impeded vision and was frequently traumatized by forelimb movements of the turtle. Twenty six months after initial presentation, the turtle was anesthetized and the bulk of the mass was surgically removed. Histopathologic examination determined the mass to be a benign lacrimal cystadenoma. PMID- 18432104 TI - Chronic endometritis in an Asian elephant (Elephas maximus). AB - A 48-yr-old female Asian elephant with a history of pododermatitis developed recurrent hematuria beginning in 2002. Transrectal ultrasonography and endoscopic examination in 2004 identified the uterus as the source of hematuria and excluded hemorrhagic cystitis. Treatment with Desloreline implants, antibiotics, and homeopathic drugs led to an improved general condition of the elephant. In July 2005, the elephant was suddenly found dead. During necropsy, the severely enlarged uterus contained about 250 L of purulent fluid, and histopathology revealed ulcerative suppurative endometritis with high numbers of Streptococcus equi ssp. zooepidemicus and Escherichia coli identified on aerobic culture. Additional findings at necropsy included: multifocal severe pododermatitis, uterine leiomyoma, and numerous large calcified areas of abdominal fat necrosis. Microbiologic culture of the pododermatitis lesion revealed the presence of Streptococcus agalactiae, Streptococcus equi ssp. zooepidemicus, Staphylococcus sp., Corynebacterium sp., and Entercoccus sp. PMID- 18432105 TI - Osteochondrosis dissecans in a juvenile roan antelope (Hippotragus equinus). AB - A 4-mo-old, juvenile roan antelope (Hippotragus equinus) presented for an acute and progressive lameness of the left hind limb, which was non-responsive to anti inflammatory medication. The antelope was anesthetized for examination, radiography, and lab work. The left stifle joint had limited range of motion and was markedly swollen on physical examination. Radiography revealed thin cortices of the left distal femur and proximal tibia, as well as multiple lytic areas in the condyles. Euthanasia was performed due to the severity of the lesions and the animal's overall physical condition. Postmortem findings included a parasitic infection of the gastrointestinal tract. Cartilaginous erosions, associated with fibrotic connective tissue and viscous joint fluid, were identified in all appendicular joints. Perimortem lab work demonstrated anemia and hypoproteinemia. Joint fluid was inflammatory but non-infectious, and aerobic culture of the joint was negative for pathogens. Histologic examination identified focal, transmural necrosis of the articular cartilage with clefting. Osteochondrosis dissecans was diagnosed, which initiated an evaluation of the nutritional status and management of the herd. This is the first documented case of osteochondrosis dissecans in a roan antelope. PMID- 18432106 TI - Use of a sustained release preparation of clotrimazole to treat dermatophytosis in a siamang (Hylobates syndactylus). AB - In November 2004, an adult male siamang (Hylobates syndactylus) from The Tisch Family Zoological Gardens-Jerusalem Biblical Zoo (Israel) presented with skin lesions on various body parts. Lesions consisted of alopecia and dry, crusty areas of hyperkeratosis. A diagnosis of dermatophytosis due to Microsporum canis was determined by fungal culture of skin scraping taken from the edge of several lesions. Treatment with various oral and topical antifungal agents such as griseofluvin, itraconozole, and lufenuron resulted in the resolution of most lesions and a decrease in size of the single remaining lesion, which continued to be culture positive for M. canis. The animal was anesthetized and an experimental sustained-release clotrimazole varnish was painted directly on the lesion. Initially there was no change in the lesion, and 2 months later a slightly altered formula was applied under anesthesia. One month later, the lesion began to reduce in size; 3 months after the start of treatment, although 2 years after the onset of clinical signs, the lesion resolved. Minimizing the number of treatments is always an advantage when dealing with exotic animals or zoological collections. PMID- 18432107 TI - Hemangiosarcoma in a South American sea lion (Otaria byronia). AB - A 10-yr-old male South American sea lion (Otaria byronia) died after several weeks of depression, anorexia, weight loss, and progressive respiratory distress. At necropsy, three confluent, lobulated, dark-red masses were noted in the mesentery. Similar masses were also observed in the lung and both kidneys. Hemangiosarcoma was diagnosed based on gross findings, histopathology, and immunohistochemistry. This is the first case of hemangiosarcoma reported in pinnipeds. PMID- 18432108 TI - Pericardial mesothelioma in a Bengal tiger (Panthera tigris). AB - A 17-year-old Bengal tiger (Panthera tigris) presented with dyspnea and tachypnea. Radiographs revealed severe pleural and pericardial effusion, but no obvious mass. During attempts to remove the fluid under anesthesia, the cat developed cardiac tamponade and died. At necropsy, a nodular mass was found at the heart base and was identified as a pericardial mesothelioma. This is the first report of this tumor in any large cat. PMID- 18432109 TI - Experimental superinfection of a Lesser Malayan mousedeer (Tragulus javanicus) persistently infected with bovine viral diarrhea virus. AB - A Lesser Malayan mousedeer (Tragulus javanicus), persistently infected with noncytopathogenic bovine viral diarrhea virus (BVDV) type 1f, was experimentally superinfected with a cytopathogenic isolate of BVDV type 1c, which antigenically partially matched the endogenous strain. Within the observational period of 125 days after superinfection, the animal did not demonstrate any clinical signs of the disease and/or significant changes in blood values. Neutralizing antibodies were detected at 35 and 42 days postinfection. The isolate causing the superinfection was found in feces, nasal swabs, and saliva starting from day 29 and at various times postchallenge. Macroscopic or histologic examination did not reveal mucosal disease-like lesions, despite the detection of the cytopathogenic isolate in the salivary gland, rumen, abomasum, kidney, and superficial prescapular lymph node. Results indicate that the cytopathogenic BVDV strain, which was used in the superinfection, persisted in the viremic animal without causing disease within the observation period. PMID- 18432110 TI - Molecular epidemiological studies of Vibrio cholerae in Bengal region. AB - Vibrio cholerae isolates from environmental and clinical origins in the Bengal region in which epidemics of cholera break out periodically were analyzed with particular emphasis on the molecular epidemiological features. The presence of the virulence genes (ctxA, tcpA and toxR) in the isolates was analyzed by the PCR (polymerase chain reaction) method. PFGE (pulsed-field gel electrophoresis) was performed to determine the clonal relationships between the clinical and environmental strains. Antibiograms and O serovars of the isolates were also examined. O1 and O139 strains from both clinical and environmental sources were all positive for the three virulence genes while non-O1/non-O139 strains from both sources were all negative for ctxA and tcpA but positive for toxR. PFGE patterns of recent isolates of O1 and O139 were similar in each serovar regardless of origin, suggesting a clonal relationship between the clinical and environmental strains, although comparison with past isolates or isolates from different geographical area showed some differences. PMID- 18432111 TI - Cleanability of titanium and stainless steel particles in relation to surface charge aspects. AB - The cleanability of titanium and 316L stainless steel particles was studied in terms of their apparent surface charge density (sigma(app)). Bovine serum albumin (BSA) was used as the model fouling agent. Curves for the sigma(app) of titanium and stainless steel particles showed the apparent points of zero charge (pzc(app)) of 4.6 and 8.5, respectively. Compared with the curve for the sigma(app) of stainless steel, that of titanium was characterized by small positive and large negative sigma(app) values. The isotherms for BSA adsorption and the saturation amount of BSA adsorbed on titanium and stainless steel depended largely on the intrinsic properties of BSA. In continuous cleaning in a plug-flow column fed by a 0.05M NaOH solution, BSA was found to be faster desorbed from titanium than from stainless steel, and smaller amounts of BSA remaining after 120-min cleaning were observed on titanium. Kinetic analysis showed that the two first-order desorption rate constants, reflecting the rate of BSA desorption in the initial and later stages of cleaning, for titanium were respectively 1.7-fold and 1.3-fold higher than those values for stainless steel. It could be suggested that the better cleanability of titanium was probably due to the small binding strength of BSA on slightly negatively-charged titanium surfaces and due to their large negative sigma(app) values under alkaline cleaning conditions. PMID- 18432112 TI - Phylogenetic analysis of carotenoid-producing marine microorganisms around the coral reefs of the Kerama Islands of Okinawa. AB - Seawater sample from the coral reefs of the Kerama Islands of Okinawa were assessed for the presence of carotenoid-producing bacteria. Results of 16S rDNA analysis of the bacteria obtained from the isolated bacteria showed unique patterns that were different from those of the bacteria obtained from the ordinary marine area. Phylogenetic analysis revealed a slight correlation with the statistical analysis of the PDA chart patterns. The results suggest that useful materials for human health such as carotenoids can be extracted from many carotenoid-producing bacteria such as those found the coral reefs the Kerama Islands. PMID- 18432113 TI - Antioxidant effects and antimicrobial activites of phytoncide. AB - Physiological activities of four types of phytoncide solutions (A, AB, CY and D types), prepared from various plants widely distributed in nature, were examined. We assayed these phytoncide solutions, testing for active oxygen inhibition, DPPH radical scavenging effects, nitrogen monoxide inhibition, and lipid peroxide inhibition. The AB- and D-types of phytoncide solutions especially showed comparably potent antioxidant effects. Antibacterial and antifungal assays were conducted using the AB- and D-types, with results showing significant inhibitory activities with these solutions. PMID- 18432114 TI - Inhibitory effects of bovine lactoferrin and lactoferricin B on Enterobacter sakazakii. AB - The susceptibility of Enterobacter sakazakii, a food-borne pathogen, to several metal-bound forms of bovine lactoferrin (LF), pepsin-hydrolyzed LF (LF-hyd), and LF-derived peptide lactoferricin B (LFcin B) was tested. MIC and MBC testing revealed that 4 strains of E. sakazakii show susceptibility to apo- and Cu-LF, LF hyd, and LFcin B, but not to Fe-LF, similarly to Escherichia coli. A growth curve test indicated that E. sakazakii was inhibited in a dose-dependent manner by apo LF at 0.5 to 8 mg/ml. Even after being heated at 80 degrees C, LF at above 1 mg/ml inhibited the bacterial growth. These results suggest that bovine LF related compounds may be useful for the inhibition of E. sakazakii in foods. PMID- 18432115 TI - First 14 communities designated chartered value exchanges by HHS. AB - Involving all the stakeholders in a community enhances your odds for success. Community organizations share successes, failures with other groups across the country. Groups chosen as CVEs already have impressive QI track records. PMID- 18432116 TI - Urinary tract infections overlooked by hospitals. AB - Nearly half of hospitals surveyed lacked a system that tells them which patients have a catheter. Limited morbidity, relatively low costs may be behind poor performance. A reminder system is a simple, yet effective strategy for minimizing urinary tract infections. PMID- 18432117 TI - Patient safety alert. 'Smart rooms' provide patient safety info at the bedside. PMID- 18432118 TI - Mortality, costs still declining in HQID demo. AB - Average improvement of 17.3% seen across all areas since beginning of initiative. All areas of a hospital must make quality a high priority. Reliability is a critical factor in measuring quality performance. PMID- 18432119 TI - Hand gel fails to curb infections on its own. AB - Infection rates are relatively steady even though compliance improves significantly. Proper positioning of ventilator patients, proper mouth care, prudent use of antibiotics are important strategies. Long fingernails, rings can increase the risk of infection. PMID- 18432120 TI - Joint commission releases proposed 2009 NPSGs. AB - New language added for HAIs to include MRSA and CDAD. Nurses will not be allowed to identify surgical sites. Close alignment with Medicare could cause Joint Commission to overlook important topics. PMID- 18432121 TI - [Caspases--structure and function]. AB - Caspases, a unique family of cysteine proteases, are involved in both the initiation and execution phase of cell apoptosis. They also play a vital role in the inflammatory responses. Caspases exist in cells as inactive zymogenes and undergo a cascade activation, which inevitably leads to the cell death. These highly specialized enzymes create the network of intracellular signaling through proteolysis. The network regulates the function and the life span of cells in physiological and pathological states. PMID- 18432122 TI - [Predictors of efficacy initial low energy of external rectilinear biphasic cardioversion in persistent atrial fibrillation]. AB - Rectilinear biphasic cardioversion (Bi-DC) has been shown to be very effective for restoration of sinus rhythm in patients with persistent atrial fibrillation (AF). There is, however, limited information on factors influencing effectiveness of the initial energy of 50 J for Bi-DC of atrial fibrillation. AIM OF THE STUDY: Evaluation of efficacy of 50 J shock of Bi-DC for restoration sinus rhythm and development of point score system to predict high conversion rate of 50 J shock in persistent atrial fibrillation. MATERIAL AND METHODS: The study group was composed of 502 consecutive Bi-DC in-patients with persistent atrial fibrillation who underwent cardioversion according to the standard protocol (50 J --> 1 J/kg m.c. --> 2 J/kg m.c. --> 200 J). Factors influencing effectiveness of 50 J shock were defined. RESULTS: Rectilinear biphasic cardioversion of shock of 50 J was successful in 157 (31%) patients. In multivariate analysis independent factors associated with efficacy of 50 J shock were: atrial fibrillation duration < 7 months (OR: 14.3, CI: 5.83 - 35.2, p < 0.001), left ventricular ejection fraction -LVEF > 40% (OR: 5.67, CI: 1.22 - 26.3, p = 0.027), body weight < 78 kg (OR: 3.17, CI: 1.52 - 6.59, p = 0.002), permanent pacemaker (OR: 2.98, CI: 1.20 - 7.40, p = 0.018), LA diameter < 4.5 cm (OR: 2.80, CI: 1.19 - 6.58, p = 0.02). A simplified point score system was developed to predict the chance for termination of atrial fibrillation (the score gives 5 points for atrial fibrillation duration < 7 months; 2 points for EF > 40%; 1 point for pacemaker, body weight < 78 kg and LA < 4.5 cm, each). High effectiveness (65%) of 50 J shock was achieved in patients with > 9 points of scoring system and 80% in this group when atrial fibrillation duration is shorter than 3 months. Effectiveness of 50 J shock was very low in the remaining group (0 points--0%; 1-3 points--5%; 4-6 points--21%; 7 8 points--34%, respectively). CONCLUSION: The efficacy of initial shock of 50 J for termination of atrial fibrillation is limited and is not recommended for general population of patients with atrial fibrillation referred for Bi-DC. 50 J might be considered in patients with pacemakers without factors associated with failure of 50 J to terminate atrial fibrillation: atrial fibrillation duration > 7 months, lower LVEF increased LA diameter, body weight > 78 kg. The shock of 50 J is effective in more than 60% of patients, if they achieved 9 or 10 points in proposed score. The highest efficacy of 50 J shock (80%) is possible to reach in this group if atrial fibrillation duration is shorter than 3 months. PMID- 18432123 TI - [Results of one year pulse oxymetry in patients with chronic thromboembolic pulmonary hypertension]. AB - It is little known on oxygen saturation in patients with chronic thromboembolic pulmonary hypertension (CTEPH). AIM OF THE STUDY: To assess the occurrence, evolution and clinical significance of oxygen desaturations (D) during 1-year observation in CTEPH patients. MATERIAL AND METHOD: The study involved 24 consecutive patients with CTEPH (6 males, 18 females, mean age 63, range 22-75 years). During 24-hour pulse oxymetry mean oxygen saturation (SpO2), number and duration of desaturations, defined as at least a 6% drop of SO2, below 88%, lasting a minimum of 8 s, were recorded at baseline and after 6 and 12 months of follow-up. Simultaneously echocardiographic study and blood gases analysis was performed. RESULTS: At baseline 18 of 24 (75%) CTEPH patients had desaturations. During follow-up none of nondesaturators had desaturations episodes and all baseline desaturators had desaturations episodes after 6 and 12 months. In desaturators there was trend to aggravate the number and duration of diurnal but not nocturnal desaturations episodes after 6 and 12 months. It was accompanied by trend (p = 0.05) to increase of pulmonary pressure as assessed by echocardiographic study, whereas the opposite trend to decrease of pulmonary pressure was seen in nondesaturators. CONCLUSIONS: Results of the study show that desaturations occur in most patients with CTEPH and desaturators may have worse clinical course than nondesaturators. It may suggest the need for long oxygen therapy in desaturating CTEPH patients to slow down progression of the disease. PMID- 18432124 TI - [High risk of atherosclerosis in men aged 20-39 from Lodz agglomeration]. AB - THE AIM OF THE STUDY: To estimate the incidence of atherosclerosis risk factors in young men of Lodz city because of the highest in Poland fatality rate of circulatory system diseases. MATERIAL AND METHODS: Anamnestic data on actual diseases, smoking, alcohol drinking and physical activity were achieved from 80 men, volunteers aged 20-39 years. Body weight and height, waist and hip circumference and arterial blood pressure were measured. Blood levels of lipids: total cholesterol (TCh), its fractions LDL, and HDL (LDL-Ch, HDL-Ch) ,and triglicerydes (TG), glucose, albumins, sex hormone binding globulin (SHBG), FSH, LH, total testosterone, dehydroepiandrosterone sulphate (DHEA-S) and estradiol were determined. Calculated were body mass index (BMI), waist to hip ratio (WHR), free testosterone index (FTI), free and bioactive testosterone. RESULTS: At least 3 atherosclerosis risk factors were simultaneously found in 33.7% of men, of which 22.7% were 20-29-year-old and 47.2% 30-39-year-old subjects. Elevated values of TG were found in 16.2% of men, TCh in 13.7%, LDL-Ch in 7.5% and decreased values of HDL-Ch in 6.2%. Positive significant correlations were found between WHR and TCh (R = 0.39; p = 0.01), LDL-Ch (R = 0.38; p = 0.02), TG (R = 0.41; p = 0.009). WHR negatively correlated with HDL-Ch (R = -0.31; p = 0.04). 50% of men had the excessive body weight. Obese men had abdominal type of obesity in 90%. As many as 62% of subjects had excessive systolic and 21% excessive diastolic arterial blood pressure. Blood pressure positively correlated with body weight (R = 0.51; p < 0.001), BMI (R = 0.51; p < 0.001), waist circumference (R = 0.55; p < 0.001) and WHR (R = 0.44; p < 0.001). In the whole group 35% of subjects led sitting life style and did not report any other physical activity. 57.5% of men were present or past smokers. 44% of men consumed alcohol everyday or almost everyday. FTI diminished with the advancing age, what was connected with the increase in SHBG blood concentration. There were no changes in total, free or bioactive testosterone, or LH and FSH concentrations with the age. Correlations between androgens and lipid profiles were not found. Estradiol blood levels negatively correlated with TG (R = -0.35; p = 0.03) and was significantly lower in 30-39-year-old men than in younger (20-29). CONCLUSION: The results indicate considerably higher incidence of atherosclerosis risk factors in young men, citizens of Lodz agglomeration, than it was found before for other regions of Poland. This phenomenon increases with the advancing age already between 20 and 39 years. Implementation of intensive prophylactic actions may prevent it. PMID- 18432125 TI - [The impact of the sonotherapy and compression therapy on enhancement of healing venous leg ulcers after surgical treatment]. AB - The aim of this paper was to describe the impact of the sonotherapy and compression therapy on enhancement of healing venous leg ulcers after surgical treatment. MATERIAL AND METHODS: Three comparative groups of patients--A, B and C were made at random from patients with venous leg ulcers, which had been earlier operated by modified Babcock' method. The 24 patients were included (in use of software Monte Carlo) to the group A, 25 patients to the group B and 24 patients to the group C (control group). The patients in all comparative groups were treated pharmacologically. Ulcerations at patients in group A were additionally treated in use of ultrasound. And ulcerations at patients in group B were additionally treated in use of compression therapy. The evaluated factors were to estimate the sonotherapy and compression therapy causes any changes of the surface, longest and widest dimensions, and volume of tissue defect. For all comparative groups calculated the Gilman parameter. The weekly speed of changes of surface and volume were estimated too. RESULTS: After therapy there was statistically significant decrease of whole surface and pus-covered and granulation area, longest and widest dimensions, and volume of ulcers in all groups while there was no statistically significant difference--except pus covered areas--between the groups observed. Comparison in terms of pus-covered area indicated a significant difference between all groups, in favor groups A and B than C. CONCLUSIONS: There are no special reasons for application of ultrasound and compression therapy in enhancement of healing process--however the following results must be verified on bigger population and longer patient observation. The sonotherapy and compression therapy promote only decontamination process compared with control group. PMID- 18432126 TI - [Knowledge of the term and methods of alternative medicine in the example of the patients of one bioresonance practice]. AB - The growing number of patients of various kinds of unconventional medicine poses a question as to the reason of taking up such treatments. Do such people know whether the methods of treatment they opt for are scientifically proven? Aim of the study was to test the knowledge among patients of bioresonance practice on methods of alternative medicine and to determine the break down of those patients based on their sex, age and the level of education. MATERIAL AND METHODS: 152 patients of bioresonance practice were asked to fill in an anonymous questionnaire. The respondents were to choose unconventional methods of treatment from the list of ten. Afterwards they were asked to answer 4 questions that referred to the opinion on and experience with alternative medicine. Chi square test and t-Student test were used to find statistical significance. RESULTS: The level of education is the only factor which determines the knowledge of the methods of alternative medicine. What is more, a mere third of patients realized that they used such a method. For one in four respondents it did not matter whether the method was properly tested and scientifically proven. Some 7.5% did not understand the meaning of the term 'alternative medicine' and did not differentiate such methods from conventional ones. CONCLUSIONS: There is quite a high percentage of people who do not know that the methods of treatment selected are unconventional. The second alarming fact is that the people who decide on treatment in an unconventional way are not concerned with any scientific evidence. The knowledge of the term and methods of alternative medicine increases with the education level. PMID- 18432127 TI - [Diagnostic and therapeutic problems with leucocytoclastic vasculitis--case report]. AB - Leucocytocalstic vasculitis (LCV) is a heterogeneous group of clinical syndromes characterized by common morphological picture of necrotic changes and inflammatory infiltration of arterioles, venules and capillary walls, mainly of neutrophils with nuclear fragments. Disorders of the immunological reactions together with the development of hypersensitivity to some antigens are observed in the course of disease if the antigen cannot by identified it is called idiopathic LCV. The aim of the study was to present the case of leucocytoclastic vasculitis together with epidemiology, pathophysiology, clinical symptoms and treatment. Conclusions. A case of a 36-year-old woman with a 16-year history of symmetrical macular exanthema type changes at the area of both shins and feet, finally diagnosed with leucocytoclastic vasculitis with further development of glomerulonephritis was reported. The authors conclude that in cases of unexplained hyperergic purpura it is necessary to consider vasculitis in differential diagnosis. They underline the value of good cooperation between General Practitioner and Specialists. Fast diagnosis and early treatment implementation could prevent development of systemic complications and alleviate skin problems. PMID- 18432128 TI - [Advances in engineering of blood vessels]. AB - Tissue engineered blood vessels are an attractive alternative for synthetic vascular prostheses utilized in vascular reconstructions. Continued for over 20 years in vitro and animal experiments has recently gave an opportunity to utilize this technique in clinical medicine. Satisfying results achieved with this approach in humans encourage wider use of vascular bioprosthesis in the future. PMID- 18432129 TI - [Food allergy in infant and children--symptoms, diagnostics and treatment]. AB - Food allergy is a kind of food hypersensitivity with various immunological mechanisms. IgE-dependent reactions are the most often occurring mechanisms. Till now population studies allowing to estimate the epidemiology of food allergy were not performed. It is estimated that this problem concerns about 6-8% of children and 1-2% of adult population. Clinical manifestation may be present in every organ and system. The basic approach of diagnosis is to assess causative association between food and presence of symptoms. The most valuable diagnostic method is double blind, placebo controlled food challenge. Elimination diet is the basic treatment method. PMID- 18432130 TI - [Influence of Helicobacter pylori on gastroesophageal reflux disease]. AB - Helicobacter pylori is an etiologic factor of chronic gastritis, duodenal and gastric ulcers and gastric cancer or gastric MALT lymphoma. The role of H. pylori infection in gastroesophageal reflux disease (GERD) pathogenesis is still controversial. The presence of H. pylori infection in gastric mucosa may play a protective or aggressive role. It appears that the influence of H. pylori infection on frequency and degree of gastroesophageal reflux depends on the localization of inflammation. More active in inducing changes in mucosa are strains producing VacA and CagA cytotoxins. It has also been reported that CagA(+) strains protect more effectively against GERD and its complications. Not all results of studies confirm this dependence. Also, pathogenesis of the above dependences is not completely known. Treatment of the infection causes reduction in inflammatory changes and subsidence of symptoms, and it also reduces the risk of complications such as gastric and duodenal ulcers. The influence of H. pylori eradication on the degree of gastroesophageal reflux and the incidence of de novo GERD is undetermined. PMID- 18432131 TI - [What is new about nonsteroidal antiinflamatory drugs?]. AB - NSAIDs contain nonselective cyclooksygenase inhibitors (for COX-1 and COX-2), inhibitors to the preferential COX-2, the coxibs (sulphonamides, methylsulphones, phenylacethic acid derivatives) with 1000 fold selectivities for COX-2. COX-2 enzyme isoform are constitutively expressed in normal gastric tissue in animals and humans, in the cardiovascular system, renal, central nervous system and other. COX-2 is involved in the ischemic preconditioning mechanism and sulphones have a prooxidant activity. COX-2 inhibitors increased risk for thrombotic cardiovascular events. Long-term study VIGOR, CLASS, TARGET MEDAL revealed that celecoxib, rofecoxib, lumiracoxib, etoricoxib significantly reduced the risk of major gastrointestinal effects (ulcers, perforations, bleeding) than nonselective NSAIDs, but the rates of complicated upper gastrointestinal events were similar for etoricoxib and diclofenac. Only in VIGOR trial incidence of cardiovascular events was greater. No evidence that concomitant ASA reduced risk for cardivascular events. Potential differences in cardiovascular outcomes with the selective COX-2 inhibitors may be due to differences in the drugs molecular structures, pharmacokinetics and pharmacodynamics. In the "prothrombotic environment" contribution disorders in the balance between thromboxan A2 and prostacyclin, increased aggregation plaque, hypertension, endothelial cell dysfunction, impaired angiogenesis. Moreover COX-2 may plays a crucial role in atherosclerotic plaque stability or instability. The cardiovascular risk may be dose related and depends on duration therapy, variable selectivity for COX-2. PMID- 18432132 TI - [New biological properties of neuroleptic phenothiazines]. AB - Phenothiazines are the oldest and the largest group of neuroleptic drugs. These compounds are widely used in psychiatry, especially in the treatment of schizophrenia, maniac and delusion disorders. They exhibit valuable antiemetic, antihistaminic and antitussive properties. There have appeared numerous articles for last ten years on new biological properties of phenothiazines, among them anticancer, antibacterial, antiprotozoic, antiviral, antiprionic and multidrug resistance reversal activity. Considering intensive development in the studies on the new activities of phenothiazines it seems to be necessary to organize and clarify the recent knowledge in this field. PMID- 18432133 TI - [Biological role and importance in the skin metabolism of vitamin C]. AB - Vitamins are a group of compounds indispensable for the development, normal growth and functioning of the human body. Lack of vitamins causes serious diseases for human, even though small amounts of them are required to maintain good health. Therefore there is growing interest conceding the role of vitamin C in biochemical-physiological conditions. This article reviews the role of water- soluble vitamin C in metabolic processes and discusses criteria used for recommended ingestion and presents recommendations for vitamin C intake. In the paper is discussed in detail the influence of level vitamin C (physico-chemical aspects) on the metabolism in skin. PMID- 18432134 TI - [Calcium ions, glutaminate acid, hypothalamic-pituitary-adrenal axis, calcium dependent ATP-ase as causes of oxidative damage in depression patients--Part I]. AB - Depressive disorder is still a rising and important problem in the modem world, it affects about 15% of the population. Present forms of treatment are effective in about 70% and require monthly therapy which sometimes causes side effects. Last decade studies paid attention to theories different to monoaminergic and to neurodegenerative changes mainly in the limbic system of hippocampus. In this article authors show a relationship between calcium ions, glutaminergic transduction and disfunction of hypothalamic-pituitary-adrenal (HPA) axis. They also take into account the activity of calcium dependent ATPase and its influence on overproduction of reactive oxygen species in the central neuron system (CNS). The first part of this article proves that disregulation of HPA increases the glutaminergic conduction in neurons and causes a cytoskeletal damage in the CNS. PMID- 18432135 TI - [Social aspect of clinical research in Poland]. AB - Each year more than 400 new clinical studies are registered in Poland. They gather above 50.000 of study participants. Social opinion on clinical trials is an important factor. The paper presents the review of actual opinions on clinical research in Poland. It provides the description of standards of protection of study participants, benefits and risks related to the participation in clinical research and the role of media in creating and influencing of the social perception of clinical trials. Results of conducted questionnaire studies imply that Poles correctly identify and assess the risk of participation in clinical experiments. The primary reason for the participation seams to be the possibility to help other patients, contribution to the progress of science and standards of medical care and potential benefits for other sufferers. The need of testing the safety and efficacy of the new medication in man is generally well recognized. At the same time a substantial part of the society is concerned with the possible corruption of investigators and unethical behaviour of sponsors. The social perception of clinical research in Poland is in majority of analyzed parameters not substantially different from opinions in other member states of EU. However, the medical society should be more active in influencing and changing some negative impressions. PMID- 18432136 TI - [Contemporary concept of placebo usage in clinical trials of new drugs]. AB - The placebo-controlled trials have been almost always controversial since the beginning of introduction of this method into new pharmacotherapy investigations. From the legal and ethical point of view the usage of placebo is more and more frequently criticized because it devoids the patients of standard therapy advantages. Therefore, contemporary tendency to solve of ethical placebo problem connected with clinical trials of new drugs is often--application of more safe methods of results objectifying in new therapy by comparison of the results of clinical trials in the group taking a new drug to be tested to the one treated by conventional, standard, established the best so far known, pharmacotherapy. PMID- 18432137 TI - Typeability of PowerPlex Y (Promega) profiles in selected tissue samples incubated in various environments. AB - In cases of decomposed bodies, Y chromosomal STR markers may be useful in identification of a male relative. The authors assessed typeability of PowerPlex Y (Promega) loci in post mortem tissue material stored in various environments. Kidney, spleen and pancreas specimens were collected during autopsies of five persons aged 20-30 years, whose time of death was determined within the limit of 14 hours. Tissue material was incubated at 21 degrees C and 4 degrees C in various environmental conditions. DNA was extracted by the organic method from tissue samples collected in 7-day intervals and subsequently typed using the PowerPlexY-STR kit and ABI 310. A fast decrease in the typeability rate was seen in specimens incubated in peat soil and in sand. Kidney tissue samples were typeable in all PowerPlexY-STR loci within 63 days of incubation at 4 degrees C. Faster DNA degradation was recorded in spleen and pancreas specimens. In samples with negative genotyping results, no DNA was found by fluorometric quantitation. Decomposed soft tissues are a potential material for DNA typing. PMID- 18432138 TI - [Ketone bodies in controlled intraoperative hypothermia- preliminary study]. PMID- 18432139 TI - Sudden death due to ischemic heart disease among adolescents and young adults before and after the socio-economic changes in 1989 of Hungary. AB - BACKGROUND: Ischemic heart disease (IDH) mortality is generally accepted as an indicator of socio-economic conditions. In view of the health status of future generations, sudden cardiac deaths among adolescents and young adults are a considerable focus of attention of public health specialists. In a changing social environment, the rate of sudden deaths in youths could be a sensitive indicator in the evaluation of health effects. Coronary artery anomalies, myocarditis and cardiomyopathies are the most common causes of sudden cardiac deaths in adolescents, and young adults. The aim of this study was to detect the changes in the rate of ischemic heart disease mortality among young people before and after the socio-economic change of 1989 in Hungary. MATERIAL AND METHOD: Data on ischemic mortality were collected from the years 1979-2000. Death certificates were obtained from the Office for National Statistics. RESULTS: There was a significant reduction in coronary mortality in young male adults in the age group of 20-24 years (OR=1.69, CI=1.23-2.32, p=0.00082), 25-29 years (OR=1.3, CI=1.11 1.53, p=0.00094) and 30-34 years (OR=1.37, CI=1.25-1.51, p=0.0000001) after the socio-economic change of 1989. Among young females, the mortality showed a reduction only in the age group of 20-24 years (OR=1.69, CI=1.18-2.45, p=0.039). DISCUSSION: Mortality rates of ischemic heart disease in adolescents and young adults may reflect the changes of socio-economic conditions. Early detection and treatment of youths at risk of premature IHD offers the greatest promise and an opportunity for gender-specific interventions. PMID- 18432140 TI - [Estimation of the time of death based on the measurements of the eye temperature in comparison with other body sites]. AB - A systematic two-stage study was conducted in pigs to verify the models of postmortem body temperature decrease currently employed in forensic medicine. During the investigations, temperature recordings were performed in four body sites (eyeballs, orbit soft tissues, muscles and rectums). The results of the study support the possible use of the eyeball and also the orbit soft tissues as temperature measuring sites at the early phase after death; they have narrowed the significance of rectum temperature measurements to the late stage of postmortem body temperature decrease, shown insignificant correlations between the body weight and the temperature decrease rate constant and illustrated the functional increase of the time of death estimation error as the body cools, expressed in the distinct tendency to overestimate the calculated time of death as compared to the actual one. In the second stage of the experiment, a lack of a plateau phase was demonstrated, at least from 30 min post mortem. It was also found that in the very early post mortem period, the kinetics of cooling of all the body sites studied was better described by the two-exponential model than the single exponential one. The study also showed that the weak airflow present in the experimental conditions did not practically affect the course of cooling of the investigated body sites. Eyeball temperature measurements with an infra-red laser thermometer performed during the experiment proved to be of no use for determination of the time of death. The experiments allowed for defining the so far unreported value of physiological temperature of pig eyeball as 38 degrees C. PMID- 18432141 TI - [The role of a forensic medicine expert in criminal proceedings]. AB - The author describes actual cases from his practice as a policeman. He also presents the opinions of well-known and highly valued forensic medicine experts and lawyers about the discussed criminal cases. Drawing from his longtime experience, the author analyses the cooperation between forensic medicine experts and prosecution bodies in offences against life and health. He deals mainly with the role played by a forensic medicine expert in formulating possible versions of the event and establishing a direction the investigation should follow. The author's practice shows that it is only a good cooperation with a forensic medicine expert that can ensure a correct, fast and successful investigation. PMID- 18432142 TI - [Medico-legal opinionating in cases of medical errors in declaration of death]. AB - Declaration of death is a diagnosis and it has legal consequences. While declaring death, there is a possibility of committing a medical error. The majority of irregularities in declaring an individual dead occur in emergency situations. When a physician declares death, he is released from the professional obligation of saving the individual's life. The studies were based on the results of medico-legal evaluations in criminal cases in medical errors. In case death is erroneously declared, we may face a situation of squandering the chance of saving the life of a human being. Exposure to loss of life or to severe detriment to health are possible charges here, along with failure to offer medical assistance, although a deliberate character of the offence according to article 162 of the Polish penal code renders such a legal qualification dubious in case of medical errors. In both cases verification is needed whether an interest protected by law was still in existence, i.e. whether the individual was still alive when the physician incorrectly declared him dead. PMID- 18432143 TI - [Today and tomorrow of forensic thanatology]. AB - The essence of post-mortem examination and its special place among other disciplines of forensic medicine were presented. Attention was drawn to the role of better understanding of phenomena occurring during and directly after death in the development of forensic medicine. Progress in determination of the time of death and in estimation of the intravital character of injuries found on the corpse was discussed. The need for a wider application of results of bio- and thanatochemical analyses in determination of the cause of death and its mechanism was emphasized. Examples of progress in this domain were presented, based on a number of papers published in this field, especially in Poland. Finally, the author pointed to the necessity of upgrading training of medical doctors in the field of thanatology. PMID- 18432144 TI - [Poisonings with anticoagulant rodenticides]. AB - Anticoagulant rodenticides are commonly used in extermination of rodents. Failure to adhere to safety principles and sometimes a deliberate use of these compounds may be a reason of severe poisonings in human. Diagnosis is based on a characteristic clinical course of the disease and postmortem examinations. PMID- 18432145 TI - [Suspected azodicarbonamide poisoning in a patient with acute hemorrhaging pancreatitis]. AB - The report describes the case of death of a 22-year old man due to acute hemorrhaging pancreatitis. Azodicarbonamide poisoning was suspected in this patient. Autopsy results, data from the literature and the knowledge about mechanisms of azodicarbonamide toxicity allowed for excluding poisoning with this substance as the cause of death. PMID- 18432146 TI - [The O. J. Simpson case. Guilty or innocent?]. AB - On October 3, 1995, one hundred million of Americans gathered at their TV sets to learn the verdict in the ex-football player O. J. Simpson case. The verdict was the consequence of a skilful defense and numerous mistakes made by prosecution. Despite copious evidence, including DNA test results indicating the guilt of thedefendant, the jury decided that O. J. Simpson was not guilty. The aim of this paper is to present the unique character of this trial and defense strategy used by a group of six attorneys, who worked on that case. PMID- 18432147 TI - A three-fragment phenotype in the D16S309 locus. AB - An allele with internal restriction site or another kind of mutation has been found in D16S309 locus. Using a probe MS205 in combination with the enzyme Hinf I 3-fragment, a pattern was observed in a woman from the Upper Silesia population- the mother of a child involved in paternity testing. DNA from this woman was additionally digested with Alu I and Mbo I enzymes. All the digested samples showed a 3-fragment pattern with the probe MS205, while other probe/enzyme combinations detected only the expected phenotypes. The disputed child inherited a second, normal allele from its mother. PMID- 18432148 TI - The creation of the medical consumer. PMID- 18432149 TI - Who will take care of you when you retire? PMID- 18432150 TI - The state of South Dakota's child: 2007, part II--death during childhood. AB - South Dakota's rate of death for children 1 to 14 years of age (39 per 100,000 population) exceeds that of the nation (20 per 100,000 population), and in 2004 (the most recent year for which data are available) it is the highest rate reported for any state in the nation. An examination of these deaths shows over half are caused by injuries. American Indians comprise 15 percent of the state's population of children, yet 35 percent of all its childhood deaths are American Indian. Further, the rate of childhood death due to homicide and suicide is five and four times higher for American Indian than white children in the state. Use of community-based mortality review committees and attention to the systemic social variables that affect behavior are suggested as approaches to responding to the tragic loss of life during childhood. PMID- 18432151 TI - Idiopathic toe-walking. AB - Idiopathic toe-walking is a diagnosis of exclusion when a child presents with bilateral toe-to-toe gait. Although toe-walking is considered part of the normal gait spectrum in development, it is abnormal when persisting past the age of two. Toe-walking may be caused by cerebral palsy, congenital contracture of the Achilles tendon or paralytic muscular disorders such as Duchenne Muscular Dystrophy. Idiopathic toe-walking may be associated with developmental disorders such as autism or other myopathic or neuropathic disorders. The majority of disorders causing toe-walking can be ruled out through the history and physical examination, resulting in a diagnosis of idiopathic toe-walking. However, it may be difficult to differentiate mild forms of cerebral palsy, specifically mild spastic diplegia, and idiopathic toe-walking. The treatment options for idiopathic toe-walking include observation, conservative methods and surgical methods. Most children can be treated in the primary care setting with either observation or conservative treatment. Patients with severe contracture of the Achilles tendon, or persistent toe-walking, may need surgical intervention. The prognosis of idiopathic toe-walking is favorable with both conservative and surgical treatment allowing children to attain normal function and range of plantarflexion. The following article provides an overview of the background information, differential diagnosis and treatment options for idiopathic toe walking. PMID- 18432152 TI - Red yeast rice and omega-3 fatty acids for cholesterol management. PMID- 18432153 TI - Reflections on the "network". PMID- 18432154 TI - Depression without a cause. PMID- 18432155 TI - Medicare demonstrations--electronic health records demonstration. PMID- 18432156 TI - Response-guided therapy: optimizing treatment now and in the future. PMID- 18432157 TI - Hepatitis C virus therapy to date. AB - Chronic hepatitis C is a major contributor to cirrhosis and hepatocellular cancer worldwide, justifying the considerable research effort aimed at understanding the disease and refining its treatment. As a result, significant therapeutic advances have been made in the last decade, particularly with regard to the development of pegylated interferons and ribavirin. This review will discuss the physical properties, pharmacokinetics, viral kinetics and side-effect profiles of the different treatment options and how they have improved, culminating in the use of pegylated interferon and ribavirin combination therapy as the current standard of care. PMID- 18432158 TI - Optimizing outcomes in patients with hepatitis C virus genotype 1 or 4. AB - Currently, many decisions for the treatment of hepatitis C virus (HCV) are based on genotype, which is the most significant baseline predictor of response to therapy; however, it has become increasingly apparent that fixed treatment durations might not be appropriate for all patients. The use of on-treatment predictors such as rapid virological response (RVR) at week 4 and early virological response (EVR) at week 12 can be used to predict the likelihood of achieving a sustained virological response (SVR), helping to tailor treatment to the individual. Until now, EVR has been defined as achieving either undetectable HCV RNA (< 50 IU/ml) or a > 2 log drop in HCV RNA, but still detectable, at week 12. However, rates of SVR in patients achieving an EVR are heterogeneous. It has recently been suggested that by subdividing EVR into RVR (< 50 IU/ml at week 4), complete EVR (HCV RNA < 50 IU/ml at week 12) or partial EVR (HCV RNA > 2 log drop in HCV RNA but still detectable [> 50 IU/ml] at week 12), it might be possible to further improve the prediction of patients likely to achieve an SVR and may allow for tailoring of treatment duration. Genotype 1 and 4 patients achieving an RVR have high rates of SVR and may be candidates for shorter treatment duration. Patients with a complete EVR achieve high SVR rates with the current treatment duration of 48 weeks, whereas patients achieving a partial EVR have lower rates of SVR and could benefit from treatment intensification to 72 weeks. Here, we discuss the importance of baseline predictors of response and the emerging concept of response-guided therapy in genotype 1 and 4 patients. PMID- 18432159 TI - Optimizing outcomes in patients with hepatitis C virus genotype 2 or 3. AB - On-treatment predictors of response could be useful in optimizing treatment for patients with hepatitis C virus (HCV) genotype 2 or 3. Early virological response (EVR) has limited value as a predictor of response in genotype 2 or 3 patients, as it is achieved by 97% of this population. However, rapid virological response (RVR) measured at week 4 is a strong predictor of sustained virological response (SVR) in this group, and patients achieving an RVR may be suitable candidates for shorter treatment durations. Several small studies investigating the efficacy of shortened treatment durations in this population have been published; however, differences in study design have made their collective interpretation difficult. We discuss these studies, followed by a comparison of the data from ACCELERATE, the largest, randomized trial carried out to investigate abbreviated therapy in genotype 2 and 3 patients. The data confirm that RVR, and its use alongside significant baseline predictors, can assist in optimizing therapy. Patients achieving an RVR have high SVR rates and might be candidates for shorter treatment duration, particularly those displaying a low viral load at baseline; however, the need to consider the increased rate of relapse versus the benefits of abbreviated therapy must also be considered. Conversely, in patients who do not achieve an RVR there is evidence to suggest they may benefit from intensified therapy (longer therapy and/or increased doses). As in genotype 1 and 4 patients, response-guided therapy aims to optimize treatment outcomes for individuals, without compromising SVR rates. PMID- 18432160 TI - Ribavirin considerations in treatment optimization. AB - Ribavirin is a guanosine analogue that has little antiviral activity when used alone, but considerably enhances the efficacy of conventional and pegylated interferon in the treatment of hepatitis C virus (HCV). The precise mode of action of ribavirin is not fully understood; however, it is crucial for the achievement of high sustained virological response (SVR) rates by enhancing virological response and reducing relapse rates. Data from several studies have confirmed that higher initial doses of ribavirin lead to higher SVR rates. Furthermore, intensified ribavirin dosing might also improve SVR rates in 'difficult-to-cure' patients. It is also important to minimize ribavirin dose reductions to promote high SVR rates and to maintain ribavirin levels throughout treatment to prevent viral breakthrough and relapse. The pharmacokinetic profile of ribavirin reveals a long elimination half-life due to accumulation in the blood, such that its side-effect profile includes haemolytic anaemia. Therefore, finding the optimal ribavirin dose requires a balance between efficacy and its associated side effects to ensure improved patient outcomes. Here, we discuss how optimizing the ribavirin component of combined therapy for HCV is an essential part of treatment optimization. PMID- 18432161 TI - Future directions in therapy for chronic hepatitis C. AB - The development of new antiviral therapies in the treatment of hepatitis C virus (HCV) is reviewed, including a discussion of the potential advances that this treatment will bring. Data from new molecules in Phase I and II clinical trials, specifically polymerase and protease inhibitors, will be discussed. The potential for resistance has been reported when these have been used as monotherapy. However, their use in combination with pegylated interferon, particularly in the presence of ribavirin, has resulted in significant improvements in antiviral activity. Preliminary studies have confirmed that the new molecules are well tolerated and further clinical studies are underway to evaluate their efficacy. Nevertheless, because of its critical role at all stages of therapy, pegylated interferon is likely to remain the cornerstone of HCV therapy. PMID- 18432162 TI - Scintigraphic imaging of adrenal disease. AB - The nuclear medicine approach to adrenal imaging spans six decades with radiopharmaceuticals designed to take advantage of the unique characteristics of the functional divisions of the gland, substrate accumulation, hormone biosynthesis, receptor-ligand interactions and pharmacologic manipulation of mechanisms of central control. The recent introduction of hybrid imaging techniques that combine high resolution anatomy with the functional maps derived from the scintigraphic images is yet another step in the evolution of integrated imaging techniques that can be used to diagnostic advantage in the evaluation of disease of the adrenal gland. PMID- 18432163 TI - Long-term mortality in patients unsuitable for surgical revascularization undergoing elective left main coronary artery angioplasty. AB - AIM: Elective percutaneous coronary intervention (PCI) of left main coronary artery disease remains an important challenge in interventional cardiology. Nonetheless, this procedure is useful for patients with significant left main stenosis who are candidates for revascularization but unsuitable for coronary artery bypass graft. In this study the Authors sought to evaluate the safety and long-term mortality of PCI of left main coronary artery disease. Secondary endpoints were to analyse long-term mortality in various categories (patients<75 years vs patients<75 years, males vs females, drug eluting stents [DES] vs bare metal stents [BMS]). METHODS: Between January 2003 and December 2006, 131 patients who consecutively under-went PCI on left main stem were reviewed. The mean follow-up time was 14.0+/-10.8 months. Survival curves were plotted with the Kaplan-Meier method and compared with the Log-rank test. RESULTS: The Kaplan Meier curves did not show statistically significant differences in terms of all cause mortality at follow-up between protected and unprotected left main coronary disease (12% vs 14% respectively, P=0.67). In the protected left main group, there was a significantly higher use of DES compared with unprotected left main group (59% vs 43%, P=0.02). CONCLUSION: The data show that PCI for left main coronary disease is feasible, safe and with an acceptable long-term mortality rate in patients at high-surgical risk unsuitable for surgical revascularization. PMID- 18432164 TI - Pathogenesis of hypertrophic cardiomyopathy. Impact of growth factors on left ventricular anatomy. AB - AIM: The aim of this study was to evaluate the effect of insulin-like growth factor 1 (IGF1) and transforming growth factor beta-1 (TGFbeta-1) on collagen turnover, left ventricular (LV) hypertrophy and on passive diastolic function of the LV in hypertrophic cardiomyopathy (HCM). METHODS: This study group comprised 34 patients with non-dilated HCM. Procollagen I amino-terminal propeptide (PINP) and collagen I carboxy-terminal telopeptide (ICTP) were measured by radioimmunoassay. Matrix metalloproteinase 9 (MMP 9), IGF1 and TGFalfa-1 were determined by enzyme-linked immunosorbent assay. The difference in duration between transmitral forward (A) and pulmonary venous retrograde (Ar) waves, was considered as an estimate of passive diastolic function; the ratio between the peak flow velocity at rapid filling at the mitral level (E) and E' measured by tissue Doppler was considered an estimate of active diastolic function. LV mass was measured and normalized to body surface area (LVMi) by cardiac magnetic resonance imaging. RESULTS: LVMi correlates to E/E' (r=0.597, P=0.019 ) and is inversely related to A-Ar (r=0.453, P=0.015). TGFbeta-1 is directly related to active MMP 9 (r=0.439, P=0.012 ). IGF1 is directly related to PICP-ICTP (r=0.347, P=0.501), that expresses the balance between collagen I synthesis and its degradation. CONCLUSION: The study demonstrated that in HCM, LVMi influences active and passive diastolic dysfunction and that IGF1 stimulates collagen synthesis and TGFbeta-1 is related to LV hypertrophy. PMID- 18432165 TI - Differences between MIP and MPR techniques in the carotid artery stenosis degree measurement. Evaluation using multi-detector-row CT angiograph. AB - AIM: Stroke is the third leading cause of mortality in the world and the stenosis degree is considered a fundamental parameter for the definition of the therapeutic approach. With the development of Multi-Detector-Row CT (MDCTA) scanner, computed tomography has become a widely used imaging technique for categorizing carotid artery stenosis degree. The purpose of this paper is to compare two CT post-processing procedures, maximum intensity projection (MIP) and multi planar reconstruction (MPR) in order to evaluate their sensitivity and inter-technique agreement. METHODS: For the purpose 45 patients (35 males and 10 females), that underwent MDCTA for carotid artery evaluation, have been retrospectively evaluated. Data set were processed with the study group's workstations, by using MPR and MIP algorithms. Each patient was assessed for stenosis degree by using North American Symptomatic Carotid Endarterectomy Trial method. Statistic analysis was performed to determine the sensitivity of the used procedure. The Cohen kappa test was applied to assess the level of intra-observer agreement. Image quality was also evaluated. RESULTS: MPR sensitivity was 87.8% (95% confidence interval [CI] 81-94.5%) and MIP sensitivity was 91.1% (95% CI 85.2-97%). Agreement in MPR was 0.792 with a standard error (SE) of 0.066, and agreement in MIP was 0.836, with a SE of 0.072. CONCLUSION: Results of the study indicate the MIP algorithm is more sensitive than MPR. Best intra-observer agreement and image quality results were also observed in the MIP. Data suggest also that MIP should be the post-processing procedure to be utilized in the evaluation of carotid artery stenosis degree, when using MDCTA. Presence of big calcified plaque can determine difficulties in MIP evaluation of stenosis degree. PMID- 18432166 TI - Towards defining the role of drug eluting stents. PMID- 18432167 TI - Sirolimus, Tacrolimus and Zotarolimus eluting stents to treat bifurcated lesions: a 7-month clinical outcome comparison. AB - AIM: Drug eluting stents (DES) have been shown to reduce restenosis compared with bare metal stents in bifurcated lesions. The aim of this study was to evaluate the long-term clinical outcomes of patients with bifurcated lesions treated by 3 different DES. METHODS: Consecutive patients with symptomatic coronary artery disease on one bifurcated lesion with SB>2.25 mm (on visual estimation) undergoing at the Department of Cardiology of the Catholic University of Rome, Italy were screened. Patients treated with Sirolimus-eluting stent (Cypher Select; SES Group), Tacrolimus-eluting stent (Taxus-Liberte; TA Group) and Zotarolimus-eluting stent (Endeavor Driver; ZOT Group) were enrolled in the study. Clinical and angiographic characteristics of all patients were prospectively recorded. Major adverse clinical events (MACE), including death, acute myocardial infarction (MI) or target lesion revascularization (TVR) by either percutaneous coronary intervention (PCI) or coronary surgery were recorded during the follow-up. Incidence of definite or probable stent thrombosis was calculated according to the ARC criteria. RESULTS: Two hundred and forty-one consecutive patients were enrolled (89 Group CY, 98 Group TA and 54 Group EN). Length of follow-up was 235+/-60 days. Baseline clinical and angiographic characteristic were similar across the groups. The adopted technique for stent implantation was provisional stenting (73.4%), T-stenting technique (7%), crush (7%) and V-stenting (2.6%). The rate of patients finally treated with two stents was similar among groups. The cumulative rate of MACE (9% SES, 12% TA, 11% ZOT: P=0.7) and of TVR (2% SES, 9% TA, 7% ZOT) was similar among groups. No definite stent thrombosis was observed during follow-up, while 1 probable stent thrombosis was observed in TA group. CONCLUSION: The clinical outcome of bifurcated lesions using DES and mainly a technique of single stent implantation is good. In the present observational study, clinical adverse events did not differ in patients with bifurcated lesions treated by Cypher, Taxus or Endeavor stent implantation. PMID- 18432168 TI - Revascularization of unprotected left main coronary artery disease with percutaneous coronary intervention: the role of drug-eluting stents. AB - The percutaneous revascularization of left main coronary artery stenosis has until recently been reserved for patients at prohibitive surgical risk or for selected emergent cases. This adopted practice of coronary artery bypass grafting, as the standard of care for left main coronary artery stenosis, has largely occurred secondary to disappointing results with bare metal stents implanted in the left main coronary artery. However, in the current era of drug eluting stents (DES) which significantly reduce restenosis compared to bare metal stents, there has been a renewed interest in examining the role of percutaneous coronary intervention as a means of revascularization of left main disease. This article discusses recent and ongoing studies investigating the role of percutaneous intervention of left main disease, with an emphasis on the use of DES for this purpose. PMID- 18432169 TI - Percutaneous coronary intervention with everolimus-eluting stents (Xience V): systematic review and direct-indirect comparison meta-analyses with paclitaxel eluting stents (Taxus) and sirolimus-eluting stents (Cypher). AB - First-generation drug-eluting stents (DES) have brought major improvements in results of percutaneous coronary intervention (PCI). However, there is currently debate on the safety of these first-generation DES, given the potential for late stent thrombosis, especially after discontinuation of dual antiplatelet therapy. Second-generation DES, such as zotarolimus- (Endeavor) and everolimus-eluting stents (Xience V), have recently become available in the USA and/or Europe. Indeed, the Xience V stent holds the promise of superior anti-restenotic efficacy as well as long-term safety, yet there is uncertainty on its risk-benefit balance. Authors conducted a systematic review of basic science and clinical evidence on the Xience V, by thoroughly searching PubMed and online databases (updated September 2007). They also compared the clinical results of Xience V vs paclitaxel-eluting stents (Taxus) and sirolimus-eluting stents (Cypher) by means of direct and indirect comparison meta-analysis. A total of three clinical studies has been retrieved focusing on Xience V, however both most recent and important trials were still unpublished. The first trial compared Xience V vs bare-metal stents, whereas the other two randomized trials compared Xience V vs Taxus. Direct meta-analysis of Xience V vs Taxus showed that Xience V was significantly superior to Taxus in preventing binary angiographic restenosis and target lesion revascularization (P<0.05 for both). Indirect comparison between Xience V and Cypher, exploiting a recent 16-trial large meta-analysis, showed that Xience V was at least as effective as Cypher in preventing target lesion revascularization (P=0.12). Everolimus-eluting stents (Xience V) appear as a major breakthrough in coronary interventions, and superior efficacy has already been demonstrated in comparison to paclitaxel-eluting stents (Taxus). Data available to date also suggest that Xience V is at least as effective as sirolimus-eluting stents (Cypher). Whether long-term results and direct comparison to Cypher will also be favorable remains to be established by future clinical trials. PMID- 18432170 TI - Intravascular ultrasound guidance in drug-eluting stent deployment. AB - The role of routine intravascular ultrasound (IVUS) guidance during percutaneous coronary intervention (PCI) in the bare metal stent era remains controversial. The potential to reduce revascularization was reported without impacting non fatal myocardial infarction and death. Drug-eluting stents (DES) have realized improved clinical results in patients undergoing PCI but are not free of restenosis and are limited by thrombosis, particularly in more complex lesion types. Stent under-expansion, malposition, and incomplete lesion coverage have been identified as the key mechanisms of DES failure. The identification of these features on IVUS and their subsequent treatment may lead to improved clinical outcomes. Though a clinical benefit is intuitive, results from randomized trials are still needed to justify routine IVUS guided DES implantation. The Authors provide a review of the currently available evidence pertaining to the potential clinical benefit of IVUS guidance in DES implantation. PMID- 18432171 TI - DES implantation in saphenous vein and left internal mammary grafts. AB - Forty percent of patients treated with CABG need further revascularizations after 10 years mainly due to saphenous--more than arterial--graft disease. In this issue, the Authors make a critical review of current available literature on the treatment of saphenous and arterial graft disease, a subset of lesions for which a clear consensus for DES use is still lacking. The Authors examine both the positive and negative aspects of DES use in this setting. Percutaneous revascularizations with DES are feasible and safe. The antiproliferative properties of DES seem to be effective even in the treatment of bypass disease, in particular in saphenous grafts. The clinical efficacy of a treatment with DES is expressed mainly in the reduction of in-stent restenosis and, therefore, in the rates of target lesion revascularization (TLR). Moreover, the use of DES is not associated to higher rates of stent thrombosis and, in case of reintervention, recurrence rates seem to be limited. However, the benefit provided by DES in prevention of restenosis may be limited by the progression of the disease in other segments than those treated with stents. Percutaneous treatment of arterial bypass with DES is feasible and safe. Most of available data on DES are on anastomotic disease (data on bypass ostium and shaft are too scarce to draw any conclusion). In this case, where the use of stents is imperative, there is no evidence of advantages gained by the use of DES over BMS in terms of new revascularizations. Some unanswered questions on DES use in this setting still remain. For this reason new randomized trials are required to definitively give a reliable answer on DES efficacy in this subset of lesions. PMID- 18432172 TI - Review of percutaneous therapy for bifurcation lesions in the era of drug-eluting stents. AB - Although recent advances in percutaneous coronary interventions (PCI) have led to dramatic expansions in procedural complexity, bifurcation lesions (BL) remain a serious challenge for the interventionalist. Turbulent flow dynamics and high shear stress likely predispose coronary bifurcations to development of atherosclerotic plaques. These lesions comprise 15% to 20% of the total number of coronary interventions. When compared with non-BL interventions, BL interventions demonstrate lower procedural success rates, higher procedural costs, longer hospitalizations, and higher clinical and angiographic restenosis rates. The recent introduction of drug-eluting stents (DES) has resulted in lower incidences of target lesion/ vessel revascularization and reduction of main branch restenosis in this anatomic subset, when compared to historical bare metal stent (BMS) controls. Nonetheless, DES have not resolved the bifurcation PCI problem; and several techniques employing either 1 or 2 stents have emerged. Stenting of the main vessel with provisional side branch stenting seems to be the prevailing approach. While no definitive single BL-PCI technique has been identified, the optimal approach is likely lesion-specific. This paper reviews different treatment modalities for this complex lesion subset, with particular emphasis on the use of DES, as well as new potential therapeutic approaches. PMID- 18432173 TI - Rotational atherectomy followed by drug-eluting stent implantation (Rota-DES): a rational approach for complex calcified coronary lesions. AB - Rotational atherectomy has been regaining interest over the last couple of years after it almost has disappeared from most interventional catheterization laboratories for several years due to failure to prove its original concept of improving long term results of percutaneous coronary interventions (PCI) as was repeatedly shown in studies in the 1990s. Its revival coupled the introduction of drug-eluting stents (DES); these devices have led to treating much more complex lesions and high-risk patients by PCI. However, real-world experience suggested that off-label use of DES is associated with a higher rate of early and late stent thrombosis. Therefore, more attention is now being paid to the initial implantation technique of DES including aggressive lesion preparation to facilitate stent delivery and expansion. The limited studies with rot-ablation and DES showed promising results with no long term safety concerns. In these studies, a subtle observation was made suggesting that rot-ablation prior to DES implantation in such lesions may have an add-on effect on long term outcome compared to DES alone. An ongoing multicenter study is investigating such effect among complex calcified coronary lesions. Even if this additive benefit does not prove true, rot-ablation remains an efficient tool for preparing certain lesions to facilitate effective and safe DES implantation. Therefore, interventional training programs should focus on this difficult technique to bridge the gap of experience which resulted from neglecting it for several years. In this regard, dedicated courses at experienced sites as well as medical simulation may be appropriate. PMID- 18432174 TI - Drug-eluting stents in patients with end-stage renal disease. AB - Advanced coronary artery disease (CAD) and acute cardiac events are the most common causes of death in patients with end-stage renal disease (ESRD). However, these patients are almost always excluded from trials examining innovations in medical and revascularization strategies for coronary disease. Extrapolation of trial conclusions regarding this high-risk patient population can be misleading because the risk-benefit ratios of various interventions are markedly different from those noted in patients with normal or mildly abnormal renal function. Because of their heightened risk, ESRD patients are frequently referred for coronary revascularization, despite the absence of solid evidence to demonstrate improved survival or reduction in clinical events. The introduction and utilization of drug-eluting stents (DES) resulted in dramatic reductions in target vessel revascularization, which now challenges the traditional algorithms of clinical decisions of percutaneous vs surgical revascularization. The utilization of DES may have out-paced the clinical trial evidence of efficacy and safety, but practicing cardiologists appear to have adopted this innovation, particularly for high-risk patients. Patients with ESRD are among several subgroups of patients in whom DES utilization appears promising, although there is no definitive randomized clinical trial evidence to support this practice. This article reviews the data available in the literature on prevalence of CAD and its impact on ESRD patients, the difficulties of referring these patients for coronary revascularization, and the potential role of adding DES to the available therapeutic options. PMID- 18432175 TI - Drug-eluting stent thrombosis. AB - When compared to bare metal stents (BMS), drug-eluting stents (DES) are associated with a dramatic reduction in restenosis and target lesion revascularization. However, the benefit of DES is limited to restenosis, and DES utilization does not translate into reductions in death or myocardial infarction. Additionally, concern exists regarding the long-term safety of DES, as there appears to be a small but real increase in late (LST) and very late stent thrombosis (VLST), seen particularly after the discontinuation of antiplatelet therapy. The specter of LST and VLST has curtailed enthusiasm for widespread DES utilization mandating critical appraisal of DES and the optimal role they play in percutaneous coronary intervention. The incidence of DES thrombosis is debated and varies somewhat by definition. The mechanisms are multifactorial, and involve patient, lesion, stent and physician related factors. Some of these factors are modifiable at the physician-patient level, while others are not. This review focuses on DES thrombosis, with particular attention paid to the definitions, incidence, mechanisms and clinical implications. PMID- 18432176 TI - Antiplatelet therapy following drug-eluting stent implantation: new clinical data and recommendations. AB - Technological developments in percutaneous coronary interventions (PCI) allow the possibility for less invasive revascularization in an increasing number of patients with atherosclerotic coronary artery disease. Bare-metal stents (BMS) have considerably improved the efficacy of PCI in addition to greatly reducing restenosis. However, even with standard stents, restenosis has remained a significant limitation of this revascularization technique. The advent of drug eluting stents (DES) has dramatically reduced in-stent restenosis and, as a result, the need for repeat revascularization. However, their potential thrombogenicity has raised concerns about their clinical utility and long-term safety. Indeed, there is a possible higher rate of late stent thrombosis (LST) with DES compared with BMS. Antiplatelet therapy has been shown to be efficient in preventing DES thrombosis. Nevertheless, in the future, significant improvement will occur to improve the safety and efficacy of this therapy. This article will summarize the pathophysiology and the epidemiology of stent thrombosis (ST). Definitions of definite, probable and possible ST will be described. Furthermore, clinical risk factors for ST will be clearly enumerated. Then, the various antiplatelet therapeutic strategies used to prevent ST will be taken in consideration. Finally, a summary of the major recommendations about antiplatelet therapy made by some of the most prestigious learned societies will be presented. PMID- 18432177 TI - Future stent drug delivery systems. AB - Drug-eluting stents (DES) with antiproliferative drugs attached via polymers on the stent surface have reduced in-stent restenosis and repeat revascularization compared with bare metal stent (BMS) across nearly all lesion and patient subsets. However, the small number of patients with in-stent restenosis after DES treatment still exists. Furthermore, concerns about long-term safety of DES are raised, particularly regarding the higher-than-expected late-event thrombosis. There is no doubt that the DES will continue to play a pivotal role in the treatment of coronary artery disease, yet future designs need to incorporate features that reduce thrombosis and promote endothelialization along with maintaining the efficacy. This review focuses on novel generation of DES, discussing new programs, including new antiproliferative agents, novel polymeric and non polymeric stents. PMID- 18432178 TI - Percutaneous unprotected left main angioplasty with drug-eluting stents in a nonagenarian: feasible and safe despite recurrent restenosis. AB - The management of severe coronary artery disease in the very elderly is challenging and the most appropriate revascularization strategy highly debated. To date, coronary artery bypass surgery has been the treatment of choice for significant stenosis of unprotected left main (ULM) coronary. Whether this holds true also in very old patients is unknown. The Authors hereby report the case of a 91-year-old diabetic man who underwent coronary angiography in June 2005 for subacute unstable angina. Angiography disclosed a critical stenoses in the distal unprotected left main, left anterior descending, and distal left circumflex, which were treated with a paclitaxel-eluting stent and a bare-metal stent (BMS), respectively. Four months later, recurrent angina lead to a second angiography which showed critical restenosis in the left anterior descending and significant ostial stenosis in the left circumflex. Another paclitaxel-eluting stent was implanted in the left anterior descending, followed by kissing balloon in the left main, achieving a good angiographic result. Six months later angina recurred again: at angiography diffuse restenosis in the left main, left anterior descending and left circumflex was documented. A crushing technique was thus employed to deliver 2 sirolimus-eluting stents, achieving a satisfactory final result. Since then, and to date for more than 17 months since the last procedure, this nonagenarian has remained free from anginal chest. This case highlights the potentials and drawbacks of percutaneous coronary intervention for unprotected left main disease, suggesting nonetheless, that this revascularization strategy can be safely repeated even in elderly patients. PMID- 18432179 TI - Intracardiac echocardiography-guided atrial septal defect transcatheter closure of a very old surgical residual shunt. AB - Although some studies have suggested excellent long-term outcome, arrhythmias, pulmonary hypertension, and paradoxical cerebral embolism are mentioned as results of residual shunts in the long-term follow-up after surgical atrial septal defect (ASD) closure at a young age. In cases of previous patch closure, transcatheter repair of residual shunts can be problematic due both to clinical decision-making in the presence of elevated pulmonary pressure and to a very old patch. A 70-year-old woman operated for an ASD with synthetic patch closure when she was 35 years old was referred to our center because of recurrent paroxysmal atrial fibrillation, initially decompensated right heart failure with rest and exercise-induced dyspnea as results of a residual shunt and moderate pulmonary hypertension. Complete right heart catheterization confirmed a mean pulmonary pressure of about 55 mm Hg and a Qp:Qs ratio of 1.78. A mechanical intracardiac echocardiography study with a 9F 9 MHz UltraICE catheter (Boston Scientific Corp.) showed a highly echogenous interatrial patch with a very stiff appearance and a very high residual defect of 8.7 and 11.2 mm on the aortic valve plane and on the four-chamber views, respectively. An occlusion test with a compliant AGA medical balloon demonstrated a decrease in mean pulmonary pressure to 36 mm Hg. A 10 mm Amplatzer's ASD occluder was implanted after a first unsuccessful attempt due to patch stiffness. Three-month echocardiography follow-up demonstrated almost normal pulmonary pressure and only slight dilation of the right chambers. At six-month follow-up, the patient no longer experienced dyspnea. This case demonstrates that transcatheter closure of a residual shunt following surgical ASD repair can be successfully accomplished also in elderly patients with a very old patch and decompensated right heart failure: the balloon occlusion test and intracardiac echocardiography appear to be effective in the operative decision making process. PMID- 18432180 TI - Emergent coronary stent-graft placement for coronary artery aneurysm occlusion. PMID- 18432181 TI - Post-myocarditis autonomic imbalance: a possible cause of arrhythmias? PMID- 18432182 TI - Biodistribution, cellular uptake and DNA-incorporation of the 2'-fluoro stabilized 5-iodo-2'-deoxyuridine analog 5-iodo-(2-deoxy-2-fluoro-beta-D arabinofuranosyl)uracil (FIAU). AB - AIM: 5-Iodo-(2-deoxy-2-fluoro-beta-D-arabinofuranosyl) uracil (FIAU) has been used for non-invasive monitoring of gene therapy and as an antiviral agent experimentally and in patients. However, FIAU metabolism in tumor cells is largely unknown. Here, the biological characteristics of FIAU in human leukemia and lymphoma cells in vitro and in a xenotransplant severe combined immunodeficient (SCID)-mouse model were investigated. METHODS: The susceptibility of FIAU to glycosidic bond cleavage by thymidine phosphorylase (TP) and its phosphorylation by human thymidine kinase 1 (hTK1) were examined. Cellular uptake and DNA-incorporation were determined in the leukemia cell line HL60 and the lymphoma cell line DoHH2. Biodistribution, in vivo stability of FIAU and expression of proliferation marker(67)Ki and thymidylate synthase were assessed in SCID-mice bearing HL60 xenotransplants. Cellular distribution of FIAU was imaged by microautoradiography. RESULTS: FIAU proved to be stable against degradation by TP and was phosphorylated by hTK1. Significant cellular uptake in DoHH2 and in HL60 cells was observed. The majority of intracellular [(131)I]FIAU was DNA incorporated. In vivo, moderate dehalogenation of [(131)I]FIAU was observed. Biodistribution studies showed a tumor uptake of 1.8+/-0.4% ID/g after 30 min. The half-life of [(131)I]FIAU in blood was 43+/-2 min. Microautoradiography showed a modest accumulation of [(125)I]FIAU in proliferating cells of small intestine, spleen and tumor. CONCLUSION: Despite phosphorylation by the hTK, efficient incorporation into the DNA and high in vivo stability, FIAU accumulates only moderately and transiently in proliferating cells, suggesting that FIAU is probably not appropriate for imaging of proliferation. PMID- 18432183 TI - Scintimammography with high resolution dedicated breast camera and mammography in multifocal, multicentric and bilateral breast cancer detection: a comparative study. AB - AIM: The aim of this study was to compare preoperative planar scintimammography (SM) with high resolution dedicated breast camera (DBC) and conventional mammography in the detection of multifocal, multicentric and bilateral breast cancer and its impact on surgical planning. METHODS: A series of 264 consecutive patients, 232 with breast cancer and 32 with benign lesions, underwent [(99m)Tc]tetrofosmin planar SM with a newly developed DBC (LumaGEM 3200S/12K, Gamma Medica-Ideas Inc.). Scintigraphic with mammography data were compared and correlated with histopathological findings. RESULTS: At surgery, ipsilateral multifocal or multicentric breast cancer in 40 patients, invasive in 24 cases (group 1) and in situ in 16 (group 2) was ascertained, and synchronous bilateral breast cancer in 4 patients (group 3). Globally, SM was positive for cancer in 41 out of the 44 breast cancer patients (93.2%) and mammography was positive in 40 out of 44 (90.1%). SM revealed multifocal/multicentric disease in 87.5% of group 1/goup 2 patients and mammography in 47.5% (P<0.0005): SM detected a significantly higher number of additional invasive foci than mammography (89.6% vs 37.9%, P<0.0005); only SM revealed ipsilateral multifocality/multicentricity in 35% of cases. Bilaterality was detected by SM in 100% of group 3 patients and by mammography in 75%. Overall specificity was 88.2% for SM and 52.9% for mammography. SM correctly changed surgical management in 16% of cases. CONCLUSIONS: DBC planar SM proved a more highly sensitive diagnostic method than mammography in the preoperative assessment of both ipsilateral multifocal/multicentric breast cancer and synchronous bilateral breast cancer. The procedure contributed to correctly changing patient surgical management in some cases, suggesting its wider use complementary to mammography. PMID- 18432184 TI - Growth restriction before or after birth reduces nephron number and increases blood pressure in male rats. AB - Impaired growth in utero predicts a low nephron number and high blood pressure later in life as does slowed or accelerated growth after a normal birth weight. We measured the effects of early postnatal growth restriction, with or without prenatal growth restriction, on blood pressure and nephron number in male rat offspring. Bilateral uterine artery and vein ligation were performed to induce uteroplacental insufficiency (Restricted) on day 18 of pregnancy. Postnatal growth restriction was induced in a subset of sham operated control animals by reducing the number of pups at birth to that of the Restricted group (Reduced Litter). Compared to Controls, Restricted pups were born smaller while Reduced Litter pups weighed less by postnatal day 3 and both groups remained lighter throughout lactation. By 10 weeks of age all animals were of similar weight but the Reduced Litter rats had elevated blood pressure. At 22 weeks, Restricted but not Reduced Litter offspring were smaller and the blood pressure was increased in both groups. Restricted and Reduced Litter groups had fewer glomeruli and greater left ventricular mass than Controls. These results suggest that restriction of both perinatal and early postnatal growth increase blood pressure in male offspring. This study also demonstrates that the early postnatal period is a critical time for nephron endowment in the rat. PMID- 18432185 TI - The prevalence of hematologic and metabolic abnormalities during chronic kidney disease stages in different ethnic groups. AB - We conducted an observational cross-sectional study to determine if the prevalence of hematologic and metabolic abnormalities in chronic kidney disease (CKD) varied in different ethnic groups. We used a CKD provincial database where a complete data set at the time of registration was available as well as an estimated glomerular filtration rate (eGFR), which showed using the abbreviated MDRD formula that the patients had CKD of stages 3-5. We included patients with self-reported race of Caucasian, Oriental Asian, or South Asian. Primary outcomes were the prevalence of at least one of the following: anemia, hypocalcemia, hyperphosphatemia, hyperparathyroidism, hypoalbuminemia, and three or more laboratory abnormalities. All definitions were consistent with K/DOQI guidelines. When compared with Caucasians, Oriental Asians and South Asians had a higher prevalence of many of the metabolic abnormalities during most stages of CKD and were more likely to have any abnormality at all levels of eGFR. The prevalence of three or more laboratory abnormalities was higher in Oriental Asians at all stages and in South Asians at some levels of eGFR. These results were unchanged or exaggerated when controlled for age, gender, diabetes, and a primary diagnosis of renal disease. Hence, it appears that South Asians and Oriental Asians have more laboratory abnormalities compared with Caucasians at most levels of eGFR. PMID- 18432186 TI - Modulation of melanocortin signaling ameliorates uremic cachexia. AB - Insulin-like growth factor (IGF)-I increases muscle mass while myostatin inhibits its development. Muscle wasting is common in patients with uremic cachexia and may be due to imbalance of this regulation. We had proposed a central mechanism involving leptin and melanocortin signaling in the pathogenesis of uremic cachexia since agouti-related peptide (AgRP), a melanocortin-4 receptor antagonist, reduced uremic cachexia. Here we found that injection of AgRP into the cerebral ventricles resulted in a gain of body mass and improved metabolic rate regulation in a mouse model of uremic cachexia. These salutary effects occurred independent of increased protein and calorie intake. Myostatin mRNA and protein concentrations were increased while those of IGF-I were decreased in the skeletal muscle of uremic mice. AgRP treatment partially corrected these uremia induced changes. Suppressor of cytokine signaling-2 gene expression (SOCS2) was significantly increased in uremic animals and AgRP reduced this expression. We suggest that AgRP improves uremic cachexia and muscle wasting by a peripheral mechanism involving the balance between myostatin and IGF-I. PMID- 18432187 TI - A 56-year-old woman with sarcoidosis and acute renal failure. PMID- 18432188 TI - Common genetic variants in the chromogranin A promoter alter autonomic activity and blood pressure. AB - Chromogranin A (CHGA) is stored and released from the same secretory vesicles that contain catecholamines in chromaffin cells and noradrenergic neurons. We had previously identified common genetic variants at the CHGA locus in several human populations. Here we focus on whether inter-individual variants in the promoter region are of physiological significance. A common haplotype, CGATA (Hap-B), blunted the blood pressure response to cold stress and the effect exhibited molecular heterosis with the greatest blood pressure change found in Hap-A/Hap-B heterozygotes. Homozygosity for three minor alleles with peak effects within the haplotype predicted lower stress-induced blood pressure changes. The G-462A variant predicted resting blood pressure in the population with higher pressures occurring in heterozygotes (heterosis). Using cells transfected with CHGA promoter-luciferase reporter constructs, the Hap-B haplotype had decreased luciferase expression compared to the TTGTC (Hap-A) haplotype under both basal conditions and after activation by pre-ganglionic stimuli. The G-462A variant altered a COUP-TF transcriptional control motif. The two alleles in transfected promoters differed in basal activity and in the responses to COUP-II-TF transactivation and to retinoic acid. In vitro findings of molecular heterosis were also noted with the transfected CHGA promoter wherein the diploid combination of the two G-462A alleles gave rise to higher luciferase expression than either allele in isolation. Our results suggest that common genetic variants in the CHGA promoter may regulate heritable changes in blood pressure. PMID- 18432189 TI - Disruption of the circadian output molecule prokineticin 2 results in anxiolytic and antidepressant-like effects in mice. AB - Disrupted circadian rhythms are strictly associated with mood disorders. The suprachiasmatic nucleus (SCN) is the master pacemaker that drives circadian rhythms in mammals. However, the underlying molecular connections of circadian rhythm and mood disorders are still poorly understood. Prokineticin 2 (PK2) is a signaling molecule that is critical for transmitting the circadian rhythms from the SCN. Previously, it is has been shown that the receptor for PK2 is expressed in virtually all of the primary SCN target areas, most of which are also involved in the mood regulation. In the current study, we investigated the role of PK2 in the regulation of anxiety and depression-related behaviors. Intracerebroventricular (ICV) infusion of PK2 increased anxiety-like behavior as assessed by light-dark box. ICV delivery of PK2 also led to increased depression like behavior in the forced swimming test. Conversely, mice lacking the PK2 gene (PK2(-/-) mice) displayed significantly reduced anxiety and depression-like behaviors. Furthermore, PK2(-/-) mice showed impaired responses to new environments in terms of locomotor activity, arousal, body temperature, and food intake. Our studies, thus, indicate that PK2 signaling plays a critical role in the stress-related traits in mice, and establish a possible molecular link between circadian rhythms and mood regulation. PMID- 18432190 TI - Activation of ERK/MAPK in the lateral amygdala of the mouse is required for acquisition of a fear-potentiated startle response. AB - There is considerable interest in examining the genes that may contribute to anxiety. We examined the function of ERK/MAPK in the acquisition of conditioned fear, as measured by fear-potentiated startle (FPS) in mice as a model for anticipatory anxiety in humans. We characterized the following for the first time in the mouse: (1) the expression of the ERK/MAPK signaling pathway components at the protein level in the lateral amygdala (LA); (2) the time course of activation of phospho-activated MAPK in the LA after fear conditioning; (3) if pharmacological inhibition of pMAPK could modulate the acquisition of FPS; (4) the cell-type specificity of pMAPK in the LA after fear conditioning. Using western blot and immunohistochemistry techniques and injecting the MEK inhibitor U0126 in the LA, we showed the following: (1) both MEK1/MEK2 and ERK1/ERK2 were co-expressed in the LA of the adult mouse brain; (2) there is a peak of pMAPK at 60 min after fear conditioning; (3) the ERK/MAPK signaling pathway activation is essential for the acquisition of an FPS response; (4) at 60 min, the pMAPK are exclusively neuronal and not glial. These results emphasize the importance of this signaling pathway in the acquisition of conditioned fear in the mouse. Given the widely held view that conditioned fear models the essential aspects of anxiety disorders, the results confirm the ERK/MAPK signaling pathway as a molecular target for the treatment of anxiety disorders in the clinic. PMID- 18432191 TI - Superconductivity at 43 K in an iron-based layered compound LaO(1-x)F(x)FeAs. AB - The iron- and nickel-based layered compounds LaOFeP (refs 1, 2) and LaONiP (ref. 3) have recently been reported to exhibit low-temperature superconducting phases with transition temperatures T(c) of 3 and 5 K, respectively. Furthermore, a large increase in the midpoint T(c) of up to approximately 26 K has been realized in the isocrystalline compound LaOFeAs on doping of fluoride ions at the O2- sites (LaO(1-x)F(x)FeAs). Experimental observations and theoretical studies suggest that these transitions are related to a magnetic instability, as is the case for most superconductors based on transition metals. In the copper-based high-temperature superconductors, as well as in LaOFeAs, an increase in T(c) is often observed as a result of carrier doping in the two-dimensional electronic structure through ion substitution in the surrounding insulating layers, suggesting that the application of external pressure should further increase T(c) by enhancing charge transfer between the insulating and conducting layers. The effects of pressure on these iron oxypnictide superconductors may be more prominent than those in the copper-based systems, because the As ion has a greater electronic polarizability, owing to the covalency of the Fe-As chemical bond, and, thus, is more compressible than the divalent O2- ion. Here we report that increasing the pressure causes a steep increase in the onset T(c) of F-doped LaOFeAs, to a maximum of approximately 43 K at approximately 4 GPa. With the exception of the copper-based high-T(c) superconductors, this is the highest T(c) reported to date. The present result, together with the great freedom available in selecting the constituents of isocrystalline materials with the general formula LnOTMPn (Ln, Y or rare-earth metal; TM, transition metal; Pn, group-V, 'pnicogen', element), indicates that the layered iron oxypnictides are promising as a new material platform for further exploration of high-temperature superconductivity. PMID- 18432192 TI - NF-kappaB links innate immunity to the hypoxic response through transcriptional regulation of HIF-1alpha. AB - The hypoxic response is an ancient stress response triggered by low ambient oxygen (O2) (ref. 1) and controlled by hypoxia-inducible transcription factor-1 (HIF-1), whose alpha subunit is rapidly degraded under normoxia but stabilized when O2-dependent prolyl hydroxylases (PHDs) that target its O2-dependent degradation domain are inhibited. Thus, the amount of HIF-1alpha, which controls genes involved in energy metabolism and angiogenesis, is regulated post translationally. Another ancient stress response is the innate immune response, regulated by several transcription factors, among which NF-kappaB plays a central role. NF-kappaB activation is controlled by IkappaB kinases (IKK), mainly IKK beta, needed for phosphorylation-induced degradation of IkappaB inhibitors in response to infection and inflammation. IKK-beta is modestly activated in hypoxic cell cultures when PHDs that attenuate its activation are inhibited. However, defining the relationship between NF-kappaB and HIF-1alpha has proven elusive. Using in vitro systems, it was reported that HIF-1alpha activates NF-kappaB, that NF-kappaB controls HIF-1alpha transcription and that HIF-1alpha activation may be concurrent with inhibition of NF-kappaB. Here we show, with the use of mice lacking IKK-beta in different cell types, that NF-kappaB is a critical transcriptional activator of HIF-1alpha and that basal NF-kappaB activity is required for HIF-1alpha protein accumulation under hypoxia in cultured cells and in the liver and brain of hypoxic animals. IKK-beta deficiency results in defective induction of HIF-1alpha target genes including vascular endothelial growth factor. IKK-beta is also essential for HIF-1alpha accumulation in macrophages experiencing a bacterial infection. Hence, IKK-beta is an important physiological contributor to the hypoxic response, linking it to innate immunity and inflammation. PMID- 18432193 TI - Draper-dependent glial phagocytic activity is mediated by Src and Syk family kinase signalling. AB - The cellular machinery promoting phagocytosis of corpses of apoptotic cells is well conserved from worms to mammals. An important component is the Caenorhabditis elegans engulfment receptor CED-1 (ref. 1) and its Drosophila orthologue, Draper. The CED-1/Draper signalling pathway is also essential for the phagocytosis of other types of 'modified self' including necrotic cells, developmentally pruned axons and dendrites, and axons undergoing Wallerian degeneration. Here we show that Drosophila Shark, a non-receptor tyrosine kinase similar to mammalian Syk and Zap-70, binds Draper through an immunoreceptor tyrosine-based activation motif (ITAM) in the Draper intracellular domain. We show that Shark activity is essential for Draper-mediated signalling events in vivo, including the recruitment of glial membranes to severed axons and the phagocytosis of axonal debris and neuronal cell corpses by glia. We also show that the Src family kinase (SFK) Src42A can markedly increase Draper phosphorylation and is essential for glial phagocytic activity. We propose that ligand-dependent Draper receptor activation initiates the Src42A-dependent tyrosine phosphorylation of Draper, the association of Shark and the activation of the Draper pathway. These Draper-Src42A-Shark interactions are strikingly similar to mammalian immunoreceptor-SFK-Syk signalling events in mammalian myeloid and lymphoid cells. Thus, Draper seems to be an ancient immunoreceptor with an extracellular domain tuned to modified self, and an intracellular domain promoting phagocytosis through an ITAM-domain-SFK-Syk-mediated signalling cascade. PMID- 18432194 TI - Human cardiovascular progenitor cells develop from a KDR+ embryonic-stem-cell derived population. AB - The functional heart is comprised of distinct mesoderm-derived lineages including cardiomyocytes, endothelial cells and vascular smooth muscle cells. Studies in the mouse embryo and the mouse embryonic stem cell differentiation model have provided evidence indicating that these three lineages develop from a common Flk 1(+) (kinase insert domain protein receptor, also known as Kdr) cardiovascular progenitor that represents one of the earliest stages in mesoderm specification to the cardiovascular lineages. To determine whether a comparable progenitor is present during human cardiogenesis, we analysed the development of the cardiovascular lineages in human embryonic stem cell differentiation cultures. Here we show that after induction with combinations of activin A, bone morphogenetic protein 4 (BMP4), basic fibroblast growth factor (bFGF, also known as FGF2), vascular endothelial growth factor (VEGF, also known as VEGFA) and dickkopf homolog 1 (DKK1) in serum-free media, human embryonic-stem-cell-derived embryoid bodies generate a KDR(low)/C-KIT(CD117)(neg) population that displays cardiac, endothelial and vascular smooth muscle potential in vitro and, after transplantation, in vivo. When plated in monolayer cultures, these KDR(low)/C KIT(neg) cells differentiate to generate populations consisting of greater than 50% contracting cardiomyocytes. Populations derived from the KDR(low)/C-KIT(neg) fraction give rise to colonies that contain all three lineages when plated in methylcellulose cultures. Results from limiting dilution studies and cell-mixing experiments support the interpretation that these colonies are clones, indicating that they develop from a cardiovascular colony-forming cell. Together, these findings identify a human cardiovascular progenitor that defines one of the earliest stages of human cardiac development. PMID- 18432195 TI - Melanopsin cells are the principal conduits for rod-cone input to non-image forming vision. AB - Rod and cone photoreceptors detect light and relay this information through a multisynaptic pathway to the brain by means of retinal ganglion cells (RGCs). These retinal outputs support not only pattern vision but also non-image-forming (NIF) functions, which include circadian photoentrainment and pupillary light reflex (PLR). In mammals, NIF functions are mediated by rods, cones and the melanopsin-containing intrinsically photosensitive retinal ganglion cells (ipRGCs). Rod-cone photoreceptors and ipRGCs are complementary in signalling light intensity for NIF functions. The ipRGCs, in addition to being directly photosensitive, also receive synaptic input from rod-cone networks. To determine how the ipRGCs relay rod-cone light information for both image-forming and non image-forming functions, we genetically ablated ipRGCs in mice. Here we show that animals lacking ipRGCs retain pattern vision but have deficits in both PLR and circadian photoentrainment that are more extensive than those observed in melanopsin knockouts. The defects in PLR and photoentrainment resemble those observed in animals that lack phototransduction in all three photoreceptor classes. These results indicate that light signals for irradiance detection are dissociated from pattern vision at the retinal ganglion cell level, and animals that cannot detect light for NIF functions are still capable of image formation. PMID- 18432196 TI - Red-shifted optogenetic excitation: a tool for fast neural control derived from Volvox carteri. AB - The introduction of two microbial opsin-based tools, channelrhodopsin-2 (ChR2) and halorhodopsin (NpHR), to neuroscience has generated interest in fast, multimodal, cell type-specific neural circuit control. Here we describe a cation conducting channelrhodopsin (VChR1) from Volvox carteri that can drive spiking at 589 nm, with excitation maximum red-shifted approximately 70 nm compared with ChR2. These results demonstrate fast photostimulation with yellow light, thereby defining a functionally distinct third category of microbial rhodopsin proteins. PMID- 18432197 TI - Light-activated channels targeted to ON bipolar cells restore visual function in retinal degeneration. AB - Genetically encoded optical neuromodulators create an opportunity for circuit specific intervention in neurological diseases. One of the diseases most amenable to this approach is retinal degeneration, where the loss of photoreceptors leads to complete blindness. To restore photosensitivity, we genetically targeted a light-activated cation channel, channelrhodopsin-2, to second-order neurons, ON bipolar cells, of degenerated retinas in vivo in the Pde6b(rd1) (also known as rd1) mouse model. In the absence of 'classical' photoreceptors, we found that ON bipolar cells that were engineered to be photosensitive induced light-evoked spiking activity in ganglion cells. The rescue of light sensitivity was selective to the ON circuits that would naturally respond to increases in brightness. Despite degeneration of the outer retina, our intervention restored transient responses and center-surround organization of ganglion cells. The resulting signals were relayed to the visual cortex and were sufficient for the animals to successfully perform optomotor behavioral tasks. PMID- 18432198 TI - Three-dimensional random access multiphoton microscopy for functional imaging of neuronal activity. AB - The dynamic ability of neuronal dendrites to shape and integrate synaptic responses is the hallmark of information processing in the brain. Effectively studying this phenomenon requires concurrent measurements at multiple sites on live neurons. Substantial progress has been made by optical imaging systems that combine confocal and multiphoton microscopy with inertia-free laser scanning. However, all of the systems developed so far restrict fast imaging to two dimensions. This severely limits the extent to which neurons can be studied, as they represent complex three-dimensional structures. Here we present a new imaging system that utilizes a unique arrangement of acousto-optic deflectors to steer a focused, ultra-fast laser beam to arbitrary locations in three dimensional space without moving the objective lens. As we demonstrate, this highly versatile random-access multiphoton microscope supports functional imaging of complex three-dimensional cellular structures such as neuronal dendrites or neural populations at acquisition rates on the order of tens of kilohertz. PMID- 18432199 TI - Cleaning up water. PMID- 18432200 TI - The race for new radiation monitors. PMID- 18432202 TI - Biomimetic materials: marine inspiration. PMID- 18432203 TI - Material mechanics: an angle on sticky films. PMID- 18432204 TI - Material witness: shrouded in mystery. PMID- 18432205 TI - Fullerides: superconductivity at the limit. PMID- 18432206 TI - Information storage: around the phase-change cycle. PMID- 18432207 TI - Main group oxides: making the transition. PMID- 18432208 TI - Polymer semiconductors: a fast mover with a bright spark. PMID- 18432209 TI - Organic ferroelectrics. AB - Ferroelectricity results from one of the most representative phase transitions in solids, and is widely used for technical applications. However, observations of ferroelectricity in organic solids have until recently been limited to well-known polymer ferroelectrics and only a few low-molecular-mass compounds. Whereas the traditional use of dipolar molecules has hardly succeeded in producing ferroelectricity in general, here we review advances in the synthesis of new organic materials with promising ferroelectric properties near room temperature, using design principles in analogy to inorganic compounds. These materials are based on non-covalent molecules formed by two or more components, in which ferroelectricity arises either from molecular displacements or from the collective transfer of electrons or protons. The principle of using multi component molecular compounds leads to a much broader design flexibility and may therefore facilitate the development of future functional organics. PMID- 18432210 TI - Progress in the prevention of chronic kidney disease in Latin America. PMID- 18432212 TI - GIST with homozygous KIT exon 11 mutations. PMID- 18432216 TI - The big ome. PMID- 18432218 TI - Superconductors redux. PMID- 18432217 TI - New sources of sex cells. PMID- 18432222 TI - Researchers should explain why they use animals. PMID- 18432223 TI - Tropical farmers need productive alternatives. PMID- 18432224 TI - Media right to report small value of antidepressants. PMID- 18432225 TI - Europe's research system must change. PMID- 18432230 TI - Metabolism: food alert. PMID- 18432231 TI - Optics: light reined in. PMID- 18432233 TI - Neuroscience: current views on odour receptors. PMID- 18432235 TI - Oceanography: bottom of the top of the world. PMID- 18432234 TI - Astrophysics: exhaust inspection. PMID- 18432236 TI - Gene transcription: two worlds merged. PMID- 18432237 TI - Plant biology: scent of a rose. PMID- 18432238 TI - Crystal structures of DNA/RNA repair enzymes AlkB and ABH2 bound to dsDNA. AB - Escherichia coli AlkB and its human homologues ABH2 and ABH3 repair DNA/RNA base lesions by using a direct oxidative dealkylation mechanism. ABH2 has the primary role of guarding mammalian genomes against 1-meA damage by repairing this lesion in double-stranded DNA (dsDNA), whereas AlkB and ABH3 preferentially repair single-stranded DNA (ssDNA) lesions and can repair damaged bases in RNA. Here we show the first crystal structures of AlkB-dsDNA and ABH2-dsDNA complexes, stabilized by a chemical cross-linking strategy. This study reveals that AlkB uses an unprecedented base-flipping mechanism to access the damaged base: it squeezes together the two bases flanking the flipped-out one to maintain the base stack, explaining the preference of AlkB for repairing ssDNA lesions over dsDNA ones. In addition, the first crystal structure of ABH2, presented here, provides a structural basis for designing inhibitors of this human DNA repair protein. PMID- 18432239 TI - The inner jet of an active galactic nucleus as revealed by a radio-to-gamma-ray outburst. AB - Blazars are the most extreme active galactic nuclei. They possess oppositely directed plasma jets emanating at near light speeds from accreting supermassive black holes. According to theoretical models, such jets are propelled by magnetic fields twisted by differential rotation of the black hole's accretion disk or inertial-frame-dragging ergosphere. The flow velocity increases outward along the jet in an acceleration and collimation zone containing a coiled magnetic field. Detailed observations of outbursts of electromagnetic radiation, for which blazars are famous, can potentially probe the zone. It has hitherto not been possible to either specify the location of the outbursts or verify the general picture of jet formation. Here we report sequences of high-resolution radio images and optical polarization measurements of the blazar BL Lacertae. The data reveal a bright feature in the jet that causes a double flare of radiation from optical frequencies to TeV gamma-ray energies, as well as a delayed outburst at radio wavelengths. We conclude that the event starts in a region with a helical magnetic field that we identify with the acceleration and collimation zone predicted by the theories. The feature brightens again when it crosses a standing shock wave corresponding to the bright 'core' seen on the images. PMID- 18432240 TI - A topological Dirac insulator in a quantum spin Hall phase. AB - When electrons are subject to a large external magnetic field, the conventional charge quantum Hall effect dictates that an electronic excitation gap is generated in the sample bulk, but metallic conduction is permitted at the boundary. Recent theoretical models suggest that certain bulk insulators with large spin-orbit interactions may also naturally support conducting topological boundary states in the quantum limit, which opens up the possibility for studying unusual quantum Hall-like phenomena in zero external magnetic fields. Bulk Bi(1 x)Sb(x) single crystals are predicted to be prime candidates for one such unusual Hall phase of matter known as the topological insulator. The hallmark of a topological insulator is the existence of metallic surface states that are higher dimensional analogues of the edge states that characterize a quantum spin Hall insulator. In addition to its interesting boundary states, the bulk of Bi(1 x)Sb(x) is predicted to exhibit three-dimensional Dirac particles, another topic of heightened current interest following the new findings in two-dimensional graphene and charge quantum Hall fractionalization observed in pure bismuth. However, despite numerous transport and magnetic measurements on the Bi(1-x)Sb(x) family since the 1960s, no direct evidence of either topological Hall states or bulk Dirac particles has been found. Here, using incident-photon-energy-modulated angle-resolved photoemission spectroscopy (IPEM-ARPES), we report the direct observation of massive Dirac particles in the bulk of Bi(0.9)Sb(0.1), locate the Kramers points at the sample's boundary and provide a comprehensive mapping of the Dirac insulator's gapless surface electron bands. These findings taken together suggest that the observed surface state on the boundary of the bulk insulator is a realization of the 'topological metal'. They also suggest that this material has potential application in developing next-generation quantum computing devices that may incorporate 'light-like' bulk carriers and spin textured surface currents. PMID- 18432241 TI - Small phonon contribution to the photoemission kink in the copper oxide superconductors. AB - Despite over two decades of intense research efforts, the origin of high temperature superconductivity in the copper oxides remains elusive. Angle resolved photoemission spectroscopy experiments have revealed a kink in the dispersion relations (energy versus wavevector) of electronic states in the copper oxides at binding energies of 50-80 meV, raising the hope that this anomaly could be a key to understanding high-temperature superconductivity. The kink is often interpreted in terms of interactions between the electrons and a bosonic field. Although there is no consensus on the nature of the bosons (or even whether a boson model is appropriate), both phonons and spin fluctuations have been proposed as possible candidates. Here we report first-principles calculations of the role of phonons and the electron-phonon interaction in the photoemission spectra of La(2 - x)Sr(x)CuO4. Our calculations within the standard formalism demonstrate that the phonon-induced renormalization of the electron energies and the Fermi velocity is almost one order of magnitude smaller than the effect observed in photoemission experiments. Therefore, our result rules out electron-phonon interaction in bulk La(2 - x)Sr(x)CuO4 as the exclusive origin of the measured kink. Our conclusions are consistent with those reached independently in a recent study of the related compound YBa2Cu3O7. PMID- 18432242 TI - Eocene/Oligocene ocean de-acidification linked to Antarctic glaciation by sea level fall. AB - One of the most dramatic perturbations to the Earth system during the past 100 million years was the rapid onset of Antarctic glaciation near the Eocene/Oligocene epoch boundary (approximately 34 million years ago). This climate transition was accompanied by a deepening of the calcite compensation depth--the ocean depth at which the rate of calcium carbonate input from surface waters equals the rate of dissolution. Changes in the global carbon cycle, rather than changes in continental configuration, have recently been proposed as the most likely root cause of Antarctic glaciation, but the mechanism linking glaciation to the deepening of calcite compensation depth remains unclear. Here we use a global biogeochemical box model to test competing hypotheses put forward to explain the Eocene/Oligocene transition. We find that, of the candidate hypotheses, only shelf to deep sea carbonate partitioning is capable of explaining the observed changes in both carbon isotope composition and calcium carbonate accumulation at the sea floor. In our simulations, glacioeustatic sea level fall associated with the growth of Antarctic ice sheets permanently reduces global calcium carbonate accumulation on the continental shelves, leading to an increase in pelagic burial via permanent deepening of the calcite compensation depth. At the same time, fresh limestones are exposed to erosion, thus temporarily increasing global river inputs of dissolved carbonate and increasing seawater delta13C. Our work sheds new light on the mechanisms linking glaciation and ocean acidity change across arguably the most important climate transition of the Cenozoic era. PMID- 18432243 TI - Hydrous silicate melt at high pressure. AB - The structure and physical properties of hydrous silicate melts and the solubility of water in melts over most of the pressure regime of Earth's mantle (up to 136 GPa) remain unknown. At low pressure (up to a few gigapascals) the solubility of water increases rapidly with increasing pressure, and water has a large influence on the solidus temperature, density, viscosity and electrical conductivity. Here we report the results of first-principles molecular dynamics simulations of hydrous MgSiO3 melt. These show that pressure has a profound influence on speciation of the water component, which changes from being dominated by hydroxyls and water molecules at low pressure to extended structures at high pressure. We link this change in structure to our finding that the water silicate system becomes increasingly ideal at high pressure: we find complete miscibility of water and silicate melt throughout almost the entire mantle pressure regime. On the basis of our results, we argue that a buoyantly stable melt at the base of the upper mantle would contain approximately 3 wt% water and have an electrical conductivity of 18 S m(-1), and should therefore be detectable by means of electromagnetic sounding. PMID- 18432244 TI - Mountain pine beetle and forest carbon feedback to climate change. AB - The mountain pine beetle (Dendroctonus ponderosae Hopkins, Coleoptera: Curculionidae, Scolytinae) is a native insect of the pine forests of western North America, and its populations periodically erupt into large-scale outbreaks. During outbreaks, the resulting widespread tree mortality reduces forest carbon uptake and increases future emissions from the decay of killed trees. The impacts of insects on forest carbon dynamics, however, are generally ignored in large scale modelling analyses. The current outbreak in British Columbia, Canada, is an order of magnitude larger in area and severity than all previous recorded outbreaks. Here we estimate that the cumulative impact of the beetle outbreak in the affected region during 2000-2020 will be 270 megatonnes (Mt) carbon (or 36 g carbon m(-2) yr(-1) on average over 374,000 km2 of forest). This impact converted the forest from a small net carbon sink to a large net carbon source both during and immediately after the outbreak. In the worst year, the impacts resulting from the beetle outbreak in British Columbia were equivalent to approximately 75% of the average annual direct forest fire emissions from all of Canada during 1959 1999. The resulting reduction in net primary production was of similar magnitude to increases observed during the 1980s and 1990s as a result of global change. Climate change has contributed to the unprecedented extent and severity of this outbreak. Insect outbreaks such as this represent an important mechanism by which climate change may undermine the ability of northern forests to take up and store atmospheric carbon, and such impacts should be accounted for in large-scale modelling analyses. PMID- 18432246 TI - Crystal structure of the lambda repressor and a model for pairwise cooperative operator binding. AB - Bacteriophage lambda has for many years been a model system for understanding mechanisms of gene regulation. A 'genetic switch' enables the phage to transition from lysogenic growth to lytic development when triggered by specific environmental conditions. The key component of the switch is the cI repressor, which binds to two sets of three operator sites on the lambda chromosome that are separated by about 2,400 base pairs (bp). A hallmark of the lambda system is the pairwise cooperativity of repressor binding. In the absence of detailed structural information, it has been difficult to understand fully how repressor molecules establish the cooperativity complex. Here we present the X-ray crystal structure of the intact lambda cI repressor dimer bound to a DNA operator site. The structure of the repressor, determined by multiple isomorphous replacement methods, reveals an unusual overall architecture that allows it to adopt a conformation that appears to facilitate pairwise cooperative binding to adjacent operator sites. PMID- 18432248 TI - Placental expression of insulin-like growth factor-I, fibroblast growth factor basic and neural cell adhesion molecule in pregnancies with small for gestational age fetuses. AB - OBJECTIVE: To investigate placental expression of insulin-like growth factor-I (IGF-I), fibroblast growth factor-basic (FGF-b) and neural cell adhesion molecule (N-CAM) regarding the pathogenesis of pregnancies with small for gestational age (SGA) fetuses. STUDY DESIGN: An immunohistochemical analysis using anti-IGF-I, anti-FGF-b and anti-N-CAM antibodies was carried out on 4% paraformaldehyde-fixed placental tissues of third trimester pregnancies complicated with SGA fetuses (n=12) and subjects exhibiting appropriately grown fetuses (n=10). Immunostaining patterns of chorionic villi and amniochorionic membranes were assessed. RESULT: IGF-I, FGF-b and N-CAM immunostainings in chorionic villi demonstrated significantly increased immunoreactivities in cytotrophoblasts of SGA cases, whereas increased IGF-I immunostaining in syncitiotrophoblasts and increased N CAM immunostaining in capillary endothelium were noted in the same group. IGF-I, FGF-b and N-CAM immunostainings in amniochorionic membranes revealed significantly decreased IGF-I immunoreactivities in extravillous trophoblasts and increased IGF-I immunoreactivities in decidual cells of SGA cases, while significantly decreased N-CAM immunoreactivities in both decidual cells and extravillous trophoblasts were noted. FGF-b immunostaining revealed no significant differences in both extravillous trophoblasts and decidual cells of SGA cases. CONCLUSION: Increased placental expression of IGF-I, FGF-b and N-CAM may act in an autocrine and/or paracrine manner to restore the impaired trophoblastic proliferation, migration and metabolism at all gestational stages by means of a positive feedback mechanism. PMID- 18432250 TI - The Mouse Tumor Biology database. AB - The laboratory mouse has long been an important tool in the study of the biology and genetics of human cancer. With the advent of genetic engineering techniques, DNA microarray analyses, tissue arrays and other large-scale, high-throughput data generating methods, the amount of data available for mouse models of cancer is growing exponentially. Tools to integrate, locate and visualize these data are crucial to aid researchers in their investigations. The Mouse Tumor Biology database (http://tumor.informatics.jax.org) seeks to address that need. PMID- 18432251 TI - Ca2+ signalling checkpoints in cancer: remodelling Ca2+ for cancer cell proliferation and survival. AB - Increases in cytosolic free Ca2+ ([Ca2+]i) represent a ubiquitous signalling mechanism that controls a variety of cellular processes, including proliferation, metabolism and gene transcription, yet under certain conditions increases in intracellular Ca2+ are cytotoxic. Thus, in using Ca2+ as a messenger, cells walk a tightrope in which [Ca2+]i is strictly maintained within defined boundaries. To adhere to these boundaries and to sustain their modified phenotype, many cancer cells remodel the expression or activity of their Ca2+ signalling apparatus. Here, we review the role of Ca2+ in promoting cell proliferation and cell death, how these processes are remodelled in cancer and the opportunities this might provide for therapeutic intervention. PMID- 18432252 TI - Wnt signalling and its impact on development and cancer. AB - The Wnt signalling pathway is an ancient system that has been highly conserved during evolution. It has a crucial role in the embryonic development of all animal species, in the regeneration of tissues in adult organisms and in many other processes. Mutations or deregulated expression of components of the Wnt pathway can induce disease, most importantly cancer. The first gene to be identified that encodes a Wnt signalling component, Int1 (integration 1), was molecularly characterized from mouse tumour cells 25 years ago. In parallel, the homologous gene Wingless in Drosophila melanogaster, which produces developmental defects in embryos, was characterized. Since then, further components of the Wnt pathway have been identified and their epistatic relationships have been defined. This article is a Timeline of crucial discoveries about the components and functions of this essential pathway. PMID- 18432253 TI - Effect of 4 weeks of aerobic or resistance exercise training on arterial stiffness, blood flow and blood pressure in pre- and stage-1 hypertensives. AB - The benefits of aerobic exercise (AE) training on blood pressure (BP) and arterial stiffness are well established, but the effects of resistance training are less well delineated. The purpose of this study was to determine the impact of resistance vs aerobic training on haemodynamics and arterial stiffness. Thirty pre- or stage-1 essential hypertensives (20 men and 10 women), not on any medications, were recruited (age: 48.2 +/- 1.3 years) and randomly assigned to 4 weeks of either resistance (RE) or AE training. Before and after training, BP, arterial stiffness (pulse wave velocity (PWV)) and vasodilatory capacity (VC) were measured. Resting systolic BP (SBP) decreased following both training modes (SBP: RE, pre 136 +/- 2.9 vs. post 132 +/- 3.4; AE, pre 141 +/- 3.8 vs. post 136 +/- 3.4 mm Hg, P = 0.005; diastolic BP: RE, pre 78 +/- 1.3 vs post 74 +/- 1.6; AE, pre 80 +/- 1.6 vs. post 77 +/- 1.7 mm Hg, P = 0.001). Central PWV increased (P = 0.0001) following RE (11 +/- 0.9-12.7 +/- 0.9 ms(-1)) but decreased after AE (12.1 +/- 0.8-11.1 +/- 0.8 m s(-1). Peripheral PWV also increased (P = 0.013) following RE (RE, pre 11.5 +/- 0.8 vs. post 12.5 +/- 0.7 ms(-1)) and decreased after AE (AE, pre 12.6 +/- 0.8 vs post 11.6 +/- 0.7 m s(-1)). The VC area under the curve (VC(AUC)) increased more with RE than that with AE (RE, pre 76 +/- 8.0 vs. post 131.1 +/- 11.6; AE, pre 82.7 +/- 8.0 vs. post 110.1 +/- 11.6 ml per min per s per 100 ml, P = 0.001). Further, peak VC (VCpeak) increased more following resistance training compared to aerobic training (RE, pre 17 +/- 1.9 vs. post 25.8 +/- 2.1; AE, pre 19.2 +/- 8.4 vs post 22.9 +/- 8.4 ml per min per s per 100 ml, P = 0.005). Although both RE and AE training decreased BP, the change in pressure may be due to different mechanisms. PMID- 18432254 TI - Should obesity be blamed for the high prevalence rates of hypertension in black South African women? AB - Hypertension is highly prevalent in South Africa, resulting in high stroke mortality rates. Since obesity is very common among South African women, it is likely that obesity contributes to the hypertension prevalence. The aims were to determine whether black African women have higher blood pressures (BPs) than Caucasian women, and whether obesity is related to their cardiovascular risk. African (N=102) and Caucasian (N=115) women, matched for age and body mass index, were included. Correlations between obesity (total body fat, abdominal obesity and peripheral fat) and cardiovascular risk markers (haemodynamic parameters, lipids, inflammatory markers, prothrombotic factors, adipokines, HOMA-IR (homoeostasis model assessment insulin resistance)) were compared between the ethnic groups (adjusted for age, smoking, alcohol and physical activity). Comparisons between low- and high-BP groups were also made for each ethnic group. Results showed that African women had higher BP (P<0.01) with increased peripheral vascular resistance. Surprisingly, African women showed significantly weaker correlations between obesity measures and cardiovascular risk markers when compared to Caucasian women (specifically systolic BP, arterial resistance, cardiac output, fibrinogen, plasminogen activator inhibitor-1, leptin and resistin). Interestingly, the latter risk markers were also not significantly different between low- and high-BP African groups. African women, however, presented significant correlations of obesity with triglycerides, C-reactive protein and HOMA that were comparable to the Caucasian women. Although urban African women have higher BP than Caucasians, their obesity levels are weakly related to traditional cardiovascular risk factors compared to Caucasian women. The results, however, suggest a link with the development of insulin resistance. PMID- 18432255 TI - CRP in non-dippers: new perspectives and old queries. PMID- 18432256 TI - Accumulation of 30 min of moderately intense physical activity is a clinically meaningful treatment to reduce systolic blood pressure in prehypertension. PMID- 18432245 TI - The draft genome of the transgenic tropical fruit tree papaya (Carica papaya Linnaeus). AB - Papaya, a fruit crop cultivated in tropical and subtropical regions, is known for its nutritional benefits and medicinal applications. Here we report a 3x draft genome sequence of 'SunUp' papaya, the first commercial virus-resistant transgenic fruit tree to be sequenced. The papaya genome is three times the size of the Arabidopsis genome, but contains fewer genes, including significantly fewer disease-resistance gene analogues. Comparison of the five sequenced genomes suggests a minimal angiosperm gene set of 13,311. A lack of recent genome duplication, atypical of other angiosperm genomes sequenced so far, may account for the smaller papaya gene number in most functional groups. Nonetheless, striking amplifications in gene number within particular functional groups suggest roles in the evolution of tree-like habit, deposition and remobilization of starch reserves, attraction of seed dispersal agents, and adaptation to tropical daylengths. Transgenesis at three locations is closely associated with chloroplast insertions into the nuclear genome, and with topoisomerase I recognition sites. Papaya offers numerous advantages as a system for fruit-tree functional genomics, and this draft genome sequence provides the foundation for revealing the basis of Carica's distinguishing morpho-physiological, medicinal and nutritional properties. PMID- 18432257 TI - Treatment of hypertensive emergencies: blood pressure cosmetics or outcome evidence? PMID- 18432258 TI - The prognostic value of the ECG in hypertension: where are we now? AB - In hypertension, the presence of left ventricular hypertrophy (LVH) is associated with increased risk of both cardiovascular morbidity and mortality. To date, the electrocardiogram (ECG) remains the cornerstone of LVH diagnosis in clinical practice because it is universally available, technically easy to perform and highly specific. In the most recent European Society of Hypertension/European Society of Cardiology guidelines for the treatment of arterial hypertension, the Sokolow-Lyon voltage criterion was recommended as part of all routine assessment of subjects with hypertension. However, the use of the ECG in the diagnosis of LVH is somewhat limited by its poor sensitivity. In this review article, we discuss the individual strength and weaknesses of the commonly used ECG criteria in diagnosing LVH. In addition, we present the latest data on the prognostic significance of ECG LVH and the survival differences conferred in different genders. In view of the recent Losartan Intervention for Endpoint Reduction in Hypertension trial, the prognostic benefit of LVH regression will also be addressed. Finally, with the wider availability of echocardiography, the role of combining both modalities to improve risk stratification in hypertension is reviewed. PMID- 18432259 TI - Immunohistochemical detection of XIAP and p63 in adenomatous hyperplasia, atypical adenomatous hyperplasia, bronchioloalveolar carcinoma and well differentiated adenocarcinoma. AB - The critical distinction of bronchioloalveolar carcinoma (BAC), well differentiated adenocarcinoma (WDAC) of lung, adenomatous hyperplasia (AH) and atypical adenomatous hyperplasia (AAH), is based on morphological criteria alone, and is therefore potentially subjective. We examined expression of two markers, X linked inhibitor of apoptosis protein (XIAP), the most potent of the inhibitor of apoptosis protein (IAP) family, and p63, a marker of bronchial reserve cells (BRC) and squamous cells, in these entities. H&E slides of 37 tissue blocks from 27 patients were reviewed and classified as AH (n=7), AAH (n=8), BAC (n=9) and WDAC (n=13). Immunostaining was performed on 4 mum sections with monoclonal anti XIAP and monoclonal anti-p63. Granular or heterogeneous cytoplasmic staining for XIAP and nuclear staining for p63 were considered positive. Neither XIAP nor p63 were detected in normal lung alveolar cells. All seven AHs were negative for XIAP and negative or focally positive for p63. All eight AAHs were positive for XIAP and displayed p63 positivity in scattered cells. All BACs displayed XIAP positivity, which ranged from focal/weak to diffuse/strong. p63 was negative in seven and focally positive in two of nine BACs. Twelve of 13 WDACs showed XIAP positivity in a similar pattern to BAC; all were negative for p63. One aberrant case diagnosed on H & E as WDAC was negative for XIAP but strongly positive for p63. Significant XIAP expression appears to be useful for distinguishing AAH from AH. Commonality of XIAP staining in AAH, BAC and WDAC supports the possibility that AAH may be a pre-malignant lesion. The rarity of p63 expression confirms previous reports and supports a nonbronchial histogenesis of these entities. In contrast, diffuse p63 staining may facilitate the identification of rare cases that may have been misclassified as alveolar in origin based on morphology but may be of BRC origin. PMID- 18432261 TI - Quantitative monitoring of cell clones carrying point mutations in the BCR-ABL tyrosine kinase domain by ligation-dependent polymerase chain reaction (LD-PCR). PMID- 18432260 TI - VTD combination therapy with bortezomib-thalidomide-dexamethasone is highly effective in advanced and refractory multiple myeloma. AB - Bortezomib (V) was combined with thalidomide (T) and dexamethasone (D) in a phase I/II trial to determine dose-limiting toxicities (DLT's) and clinical activity of the VTD regimen in 85 patients with advanced and refractory myeloma. The starting dose of V was 1.0 mg/m(2) (days 1, 4, 8, 11, every 21 day) with T added from cycle 2 at 50 mg/day, with 50 mg increments per 10 patient cohorts, to a maximum dose of 200 mg. In the absence of DLT's, the same reiteration of T dose increases was applied with a higher dose of V=1.3 mg/m(2). D was added with cycle 4 in the absence of partial response (PR). Ninety-two percent had prior autotransplants, 74% had prior T and 76% abnormal cytogenetics. MTD was reached at V=1.3 mg/m(2) and T=150 mg. Minor response (MR) was recorded in 79%, and 63% achieved PR including 22% who qualified for near-complete remission. At 4 years, 6% remain event-free and 23% alive. Both OS and EFS were significantly longer in the absence of prior T exposure and when at least MR status was attained. The MMSET/FGFR3 molecular subtype was prognostically favorable, a finding since reported for a VTD-incorporating tandem transplant trial (Total Therapy 3) for untreated patients with myeloma (BJH 2008). PMID- 18432262 TI - Is BAL useful in patients with acute myeloid leukemia admitted in ICU for severe respiratory complications? AB - In patients with hematological malignancy (HM) developing acute respiratory failure (ARF) bronchoalveolar lavage (BAL) is considered as a major diagnostic tool. However, the benefit/risk ratio of this invasive procedure is probably lower in the subset of patients with acute myeloid leukemia (AML). The study was to analyze the yield of BAL performed in HM patients (n=175) with AML or lymphoid malignancies (LM) admitted in intensive care unit (ICU) for ARF and pulmonary infiltrates. BAL was performed in 121 patients (53/73 AML patients (73%) and 68/102 LM patients (67%)) without a definite diagnosis at admission or contraindication for fiberoptic bronchoscopy. Life-threatening complications were noticed in 12/121 patients (10%). The overall diagnostic yield of BAL was 47% (25/53) in AML patients and 50% (34/68) in LM patients. A microorganism was recovered from BAL in 23% (12/53) of AML patients and 41% (28/68) of LM patients (P<0.005). BAL results induced significant therapeutic changes in 17% (9/53) of AML patients vs 35% (24/68) of LM patients (P=0.039). This study underlines the rather low diagnostic yield of BAL for infectious diagnosis and the low rate of therapeutic changes induced by its results in AML patients with ARF admitted in ICU. PMID- 18432266 TI - A "vector drain" in US gene therapy development? PMID- 18432269 TI - In utero efficacy of cystic fibrosis gene therapy: difficult studies, positive or negative. PMID- 18432270 TI - Bovine adenovirus vectored vaccine for avian influenza. PMID- 18432271 TI - Therapeutic angiogenesis gains a leg to stand on. PMID- 18432272 TI - Realizing the promise of RNAi. PMID- 18432273 TI - STAT4 but not TRAF1/C5 variants influence the risk of developing rheumatoid arthritis and systemic lupus erythematosus in Colombians. AB - The aim of this study was to determine the influence of STAT4 (rs7574865) and TRAF1/C5 (rs10818488 and rs2900180) gene polymorphisms on the risk of developing rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE) in a Colombian population. This was a case-control study in which 839 individuals with RA (N=274) and SLE (N=144) and matched healthy controls (N=421) were included. Genotyping was performed by using a polymerase chain reaction system with pre developed TaqMan allelic discrimination assay. STAT4 rs7574865T allele disclosed a significant influence on the risk of developing SLE (P=0.0005; OR 1.62, 95% CI 1.22-2.16) and RA (P=0.008; OR 1.36; 95% CI 1.08-1.71), whereas no effect on these autoimmune diseases was observed for the TRAF1/C5 polymorphisms examined. Our data strengthen STAT4 rs7574865 polymorphism as a susceptibility factor for RA and SLE and provide further evidence for a common origin of autoimmune diseases. PMID- 18432274 TI - Possible pathogenic role of Th17 cells for atopic dermatitis. AB - The critical role of IL-17 has recently been reported in a variety of conditions. Since IL-17 deeply participates in the pathogenesis of psoriasis and keratinocyte production of certain cytokines, the involvement of T helper cell 17 (Th17) in atopic dermatitis (AD) is an issue to be elucidated. To evaluate the participation of Th17 cells in AD, we successfully detected circulating lymphocytes intracellularly positive for IL-17 by flow cytometry, and the IL-17+ cell population was found exclusively in CD3+CD4+ T cells. The percentage of Th17 cells was increased in peripheral blood of AD patients and associated with severity of AD. There was a significant correlation between the percentages of IL 17+ and IFN-gamma+ cells, although percentage of Th17 cells was not closely related to Th1/Th2 balance. Immunohistochemically, IL-17+ cells infiltrated in the papillary dermis of atopic eczema more markedly in the acute than chronic lesions. Finally, IL-17 stimulated keratinocytes to produce GM-CSF, TNF-alpha, IL 8, CXCL10, and VEGF. A marked synergistic effect between IL-17 and IL-22 was observed on IL-8 production. The number of Th17 cells is increased in the peripheral blood and acute lesional skin of AD. Th17 cells may exaggerate atopic eczema. PMID- 18432276 TI - AAV-mediated gene transfer for the treatment of hemophilia B: problems and prospects. AB - Adeno-associated viral vector-mediated gene transfer of coagulation factor IX to the skeletal muscle or to liver has resulted in sustained correction of hemophilia B in mice and dogs. The two initial phase I/II AAV clinical trials for hemophilia B, delivering a factor IX cDNA to skeletal muscle or liver, showed no serious adverse events. Although the muscle trial failed to achieve a therapeutic level of factor IX in the circulation, long-term expression of clotting factor was demonstrated on muscle biopsies taken up to 3 years after vector injection. Administration of vector to liver via the hepatic artery identified a therapeutic dose, which agreed closely with the doses predicted by studies in hemophilic dogs. However, expression in human subjects lasted for only a period of weeks, followed by a gradual decline in factor IX levels accompanied by a self-limited, asymptomatic rise and fall in liver enzymes. Immune responses to vector capsid may account for this difference in outcome between humans and other species. Here we review the results from both preclinical and clinical studies of adeno associated viral vector gene transfer for hemophilia B, and the problems that have been identified and that must be overcome to achieve successful transduction and sustained expression. PMID- 18432275 TI - Lentiviral transduction of the murine lung provides efficient pseudotype and developmental stage-dependent cell-specific transgene expression. AB - Gene transfer for cystic fibrosis (CF) airway disease has been hampered by the lung's innate refractivity to pathogen infection. We hypothesized that early intervention with an integrating gene transfer vector capable of transducing the lung via the lumen may be a successful therapeutic approach. An HIV-based lentiviral vector pseudotyped with the baculovirus gp64 envelope was applied to the fetal, neonatal or adult airways. Fetal intra-amniotic administration resulted in transduction of approximately 14% of airway epithelial cells, including both ciliated and non-ciliated epithelia of the upper, mid and lower airways; there was negligible alveolar or nasal transduction. Following neonatal intra-nasal administration we observed significant transduction of the airway epithelium (approximately 11%), although mainly in the distal lung, and substantial alveolar transduction. This expression was still detectable at 1 year after application. In the adult, the majority of transduction was restricted to the alveoli. In contrast, vesicular stomatitis virus glycoprotein pseudotyped virus transduced only alveoli after adult and neonatal application and no transduction was observed after fetal administration. Repeat administration did not increase transduction levels of the conducting airway epithelia. These data demonstrate that application at early developmental stages in conjunction with an appropriately pseudotyped virus provides efficient, high-level transgene expression in the murine lung. This may provide a modality for treatment for lung disease in CF. PMID- 18432277 TI - A canine minidystrophin is functional and therapeutic in mdx mice. AB - Duchenne muscular dystrophy (DMD) is the most common and lethal genetic muscle disorder lacking a curative treatment. We wish to use the dystrophin-deficient golden retriever muscular dystrophy (GRMD) dog, a canine model of DMD, to investigate adeno-associated virus (AAV) vector-mediated minidystrophin gene therapy. The dog model is useful in evaluating vector dose requirement and immunological consequences owing to its large size and outbred nature. In this study, we have cloned and constructed a canine minidystrophin gene vector. Owing to limited availability of the GRMD dogs, here we first examined the functions and therapeutic effects of the canine minidystrophin in the mdx mouse model. We observed efficient minigene expression without cellular immune responses in mdx mice after AAV1-cMinidys vector intramuscular injection. We also observed restoration of the missing dystrophin-associated protein complex (DPC) onto the sarcolemma, including sarcoglycans and dystrobrevin, and a partial restoration of alpha-syntrophin and neural nitric oxide synthase (nNOS). In addition, minidystrophin treatment ameliorated dystrophic pathology, such as fibrosis and myofiber central nucleation (CN). CN remained minimal (<2%) after AAV injection in the neonatal mdx mice and was reduced from more than 75% to about 25% after AAV injection in adult mdx mice. Finally, in vivo cell membrane leakage test with Evans blue dye showed that the canine minidystrophin could effectively protect the myofiber plasma membrane integrity. Our results, thus, demonstrated the functionality and therapeutic potential of the canine minidystrophin and paved its way for further testing in the GRMD dog model. PMID- 18432279 TI - Numbers, biomass and cultivable diversity of microbial populations relate to depth and borehole-specific conditions in groundwater from depths of 4-450 m in Olkiluoto, Finland. AB - Microbiology, chemistry and dissolved gas in groundwater from Olkiluoto, Finland, were analysed over 3 years; samples came from 16 shallow observation tubes and boreholes from depths of 3.9-16.2 m and 14 deep boreholes from depths of 35-742 m. The average total number of cells (TNC) was 3.9 x 10(5) cells per ml in the shallow groundwater and 5.7 x 10(4) cells per ml in the deep groundwater. There was a significant correlation between the amount of biomass, analysed as ATP concentration, and TNC. ATP concentration also correlated with the stacked output of anaerobic most probable number cultivations of nitrate-, iron-, manganese- and sulphate-reducing bacteria, and acetogenic bacteria and methanogens. The numbers and biomass varied at most by approximately three orders of magnitude between boreholes, and TNC and ATP were positively related to the concentration of dissolved organic carbon. Two depth zones were found where the numbers, biomass and diversity of the microbial populations peaked. Shallow groundwater down to a depth of 16.2 m on average contained more biomass and cultivable microorganisms than did deep groundwater, except in a zone at a depth of approximately 300 m where the average biomass and number of cultivable microorganisms approached those of shallow groundwater. Starting at a depth of approximately 300 m, there were steep gradients of decreasing sulphate and increasing methane concentrations with depth; together with the peaks in biomass and sulphide concentration at this depth, these suggest that anaerobic methane oxidation may be a significant process at depth in Olkiluoto. PMID- 18432278 TI - Ex vivo adenoviral transfer of bone morphogenetic protein 12 (BMP-12) cDNA improves Achilles tendon healing in a rat model. AB - The aim of our study was to evaluate the histological and biomechanical effects of BMP-12 gene transfer on the healing of rat Achilles tendons using a new approach employing a genetically modified muscle flap. Biopsies of autologous skeletal muscle were transduced with a type-five, first-generation adenovirus carrying the human BMP-12 cDNA (Ad.BMP-12) and surgically implanted around experimentally transected Achilles tendons in a rat model. The effect of gene transfer on healing was evaluated by mechanical and histological testing after 1, 2, 4 and 8 weeks. One week after surgery, the maximum failure load of the healing tendons was significantly increased in the BMP-12 group, compared with the controls, and the tendon stiffness was significantly higher at 1, 2 and 4 weeks. Moreover, the size of the rupture callus was increased in the presence of BMP-12 and there was evidence of accelerated remodeling of the lesion in response to BMP 12. Histological examination showed a much more organized and homogeneous pattern of collagen fibers at all time points in lesions treated with the BMP-12 cDNA muscle graft. Both single fibrils and the collagen fibers had a greater diameter, with a higher degree of collagen crimp than the collagen of the control groups. This was confirmed by sirius red staining in conjunction with polarized light microscopy, which showed a higher shift of small yellow-green fibers to strong yellow-orange fibers after 2, 4 and 8 weeks in the presence of BMP-12 cDNA. There was also an earlier shift from fibroblasts to fibrocytes within the healing tendon, with less fat cells present in the tendons of the BMP-12 group compared with the controls. Treatment with BMP-12 cDNA-transduced muscle grafts thus produced a promising acceleration and improvement of tendon healing, particularly influencing early tissue regeneration, leading to quicker recovery and improved biomechanical properties of the Achilles tendon. Further development of this approach could have clinical applications. PMID- 18432280 TI - Introduction. PMID- 18432281 TI - Patenting and licensing in genetic testing: recommendations of the European Society of Human Genetics. AB - Patents for inventions can be beneficial for society, if they drive innovation and promote progress. In most areas, the patenting system works satisfactorily. However, it must be recognized that in some instances it can also be problematic; this is the case in the field of genetics, and particularly in the area of genetic testing. As patents should serve their original purpose (promoting innovation through a fair reward system for the inventors), the European Society of Human Genetics (ESHG) suggests ways to improve the mechanisms that already form part of the patents system as a whole. In brief, the ESHG recommends limiting the breadth of the claims in genetic patents and, more practically, to reduce the number of patents by limiting the patentable subject matter, thereby improving the quality of the patents that will eventually be granted. There is also a suggestion to redefine the concept of utility in patent law, by taking account of downstream clinical experience. The ESHG sees no harm in the patenting of novel technical tools for genetic testing (eg PCR or chip technologies), as they can promote investment and still allow for invention around them. Many disputes between supporters of the patenting system and the public revolve around ethical issues. The European Patent Office should consider the benefit of having an ethics committee to consider issues of major interest, such as patents applied to genes. The problem of licensing should also be addressed. Practically, this means supporting the Organisation for Economic Co-operation and Development guidelines, which prescribe that licences should be non-exclusive and easily obtainable, both in practical and in financial terms. To promote this, the practical exploration of alternative models for licensing, like patent pools and clearinghouses, is a prerequisite. To better track developments in this field, the establishment of a voluntary reporting system, whereby geneticists could report on any issues related to new and/or old patents or licences in the light of service provision to patients, would be worthwhile. Finally, the ESHG is calling upon all stakeholders to start the process of developing a code of conduct for partners with patents, covering ethical aspects as well as smooth licensing arrangements. PMID- 18432282 TI - Injury-induced expression of cytokeratins 8 and 18 by vascular smooth muscle cells requires concurrent activation of cytoskeletal and growth factor receptors. AB - Cytokeratins are not present in the vascular smooth muscle cells (VSMCs) of normal arteries, but they are detectable in the VSMCs of atherosclerotic lesions. A correlation between cytokeratin expression and VSMC phenotype is proposed, but an examination of VSMCs after mechanical injury has yet to be performed. Immunohistochemistry was used to monitor proteins in arterial sections. Western blotting enabled quantification of protein levels. Angioplasty of porcine femoral artery in vivo and porcine coronary artery in vitro served as models of vascular injury. Cytokeratins 8 and 18 were expressed by VSMCs in porcine femoral artery lesions 14 days after balloon angioplasty. Cytokeratins were also present in the neointima of porcine coronary artery segments placed into organ culture for 4 days. Cytokeratin expression was decreased in the presence of inhibitors that affect MAP kinase, PI3 kinase, Src kinase, and G protein, but not in the presence of an AT1 receptor antagonist. Cytokeratin expression also occurred when VSMCs were plated onto collagen in the presence of serum. We conclude that mechanical injury induces expression of cytokeratin 8 and 18 both in vitro and in vivo by synthetic VSMCs that migrate into the neointima. Furthermore, cytokeratin expression requires cellular attachment to extracellular matrix proteins in conjunction with mitogenic stimulation. PMID- 18432283 TI - Reduction of enantioselectivity in the kinetic disposition and metabolism of verapamil in rats exposed to toluene. AB - Toluene and verapamil are subject to extensive oxidative metabolism mediated by CYP enzymes, and their interaction can be stereoselective. In the present study we investigated the influence of toluene inhalation on the enantioselective kinetic disposition of verapamil and its metabolite, norverapamil, in rats. Male Wistar rats (n = 6 per group) received a single dose of racemic verapamil (10 mg/kg) orally at the fifth day of nose-only toluene or air (control group) inhalation for 6 h/day (25, 50, and 100 ppm). Serial blood samples were collected from the tail up to 6 h after verapamil administration. The plasma concentrations of verapamil and norverapamil enantiomers were analyzed by LC-MS/MS by using a Chiralpak AD column. Toluene inhalation did not influence the kinetic disposition of verapamil or norverapamil enantiomers (p > 0.05, Kruskal-Wallis test) in rats. The pharmacokinetics of verapamil was enantioselective in the control group, with a higher plasma proportion of the S-verapamil (AUC 250.8 versus 120.4 ng x h x mL(-1); p < or = 0.05, Wilcoxon test) and S-norverapamil (AUC 72.3 versus 52.3 ng x h x mL(-1); p < or = 0.05, Wilcoxon test). Nose-only exposure to toluene at 25, 50, or 100 ppm resulted in a lack of enantioselectivity for both verapamil and norverapamil. The study demonstrates the importance of the application of enantioselective methods in studies on the interaction between solvents and chiral drugs. PMID- 18432284 TI - Prolactin decreases expression of Runx2, osteoprotegerin, and RANKL in primary osteoblasts derived from tibiae of adult female rats. AB - Hyperprolactinemia caused by physiological or pathological conditions, such as those occurring during lactation and prolactinoma, respectively, results in progressive osteopenia. The underlying mechanisms, however, are controversial. Prolactin (PRL) may directly attenuate the functions of osteoblasts, since these bone cells express PRL receptors. The present study therefore aimed to investigate the effects of PRL on the expression of genes related to the osteoblast functions by using quantitative real-time PCR technique. Herein, we used primary osteoblasts that were derived from the tibiae of adult rats and displayed characteristics of differentiated osteoblasts, including in vitro mineralization. Osteoblasts exposed for 48 h to 1000 ng/mL PRL, but not to 10 or 100 ng/mL PRL, showed decreases in the mRNA expression of Runx2, osteoprotegerin (OPG), and receptor activator of nuclear factor kappaBeta ligand (RANKL) by 60.49%, 72.74%, and 87.51%, respectively. Nevertheless, PRL did not change the RANKL/OPG ratio, since expression of OPG and RANKL were proportionally decreased. These concentrations of PRL had no effect on the mRNA expression of osteocalcin and osteopontin, nor on mineralization. High pathologic concentrations of PRL (1000 ng/mL) may downregulate expression of genes that are essential for osteoblast differentiation and functions. The present results explained the clinical findings of hyperprolactinemia-induced bone loss. PMID- 18432285 TI - Increase in extracellular dopamine levels during clozapine-induced potentiation in the hippocampal dentate gyrus of chronically prepared rabbits. AB - We previously found that 20 mg/kg clozapine i.p. potentiated the excitatory synaptic responses elicited in the dentate gyrus by single electrical stimulation of the perforant path in chronically prepared rabbits. We called this phenomenon clozapine-induced potentiation and proved that it was an NMDA receptor-mediated event. This potentiation is presumably related to clozapine's clinical effect on negative symptoms and cognitive dysfunctions in schizophrenia. In the present study, to investigate the mechanisms underlying clozapine-induced potentiation, we examined whether extracellular dopamine and 5-HT levels changed during the potentiation by using a microdialysis technique in the dentate gyrus. The extracellular concentrations of dopamine and 5-HT levels were measured every 5 min during all experiments. Extracellular 5-HT levels did not change, but dopamine levels eventually increased significantly during clozapine-induced potentiation. The increase in the dopamine levels occurred almost simultaneously with the induction of clozapine-induced potentiation. These results suggest that clozapine-induced potentiation is at least partly attributable to a dopamine mediated potentiation of excitatory synaptic transmission. The present study implies that such phenomena occur also in the perforant path-dentate gyrus pathway. PMID- 18432286 TI - Modulation of radiosensitivity in Chinese hamster lung fibroblasts by cisplatin. AB - The effects of cisplatin exposure time, concentration, and irradiation sequence on the sensitivity of Chinese hamster lung fibroblasts (V79) to gamma-ray exposure were examined. Based on clonogenic cell survival, the cisplatin concentrations corresponding to 50% cell survival (EC(50)) for exposure times of 1 h to 7 days followed a 2-phase exponential decay and ranged from 28.26 +/- 3.32 to 1.53 +/- 0.24 micromol/L, respectively. When cells were treated at EC(50) for exposures of less than 4 h and irradiated immediately, cisplatin inhibited the effect of radiation. Exposures of 4-6 h did not affect radiosensitivity. For exposures of 8-12 h, radiosensitization was observed, which disappeared at 14 h and reappeared for much longer cisplatin treatments. At the lowest achievable EC(50) (1.53 micromol/L), radiosensitization was observed if irradiation was delayed for 1-8 h. This enhancement in radiosensitivity disappeared for irradiation delays of 10-12 h, but reappeared when irradiation was delayed for 14 18 h. These data demonstrate that the mode of interaction between cisplatin and gamma-irradiation depends on the concentration and exposure time of cisplatin, as well as on the timing of irradiation after cisplatin administration. Consideration of changes in cell cycle kinetics may contribute to the improvement of treatment outcomes in adjuvant chemoradiotherapy involving cisplatin. PMID- 18432287 TI - Ginger attenuates acetylcholine-induced contraction and Ca2+ signalling in murine airway smooth muscle cells. AB - Asthma is a chronic disease characterized by inflammation and hypersensitivity of airway smooth muscle cells (ASMCs) to different spasmogens. The past decade has seen increased use of herbal treatments for many chronic illnesses. Ginger (Zingiber officinale) is a common food plant that has been used for centuries in treating respiratory illnesses. In this study, we report the effect of its 70% aqueous methanolic crude extract (Zo.Cr) on acetylcholine (ACh)-induced airway contraction and Ca(2+) signalling in ASMCs using mouse lung slices. Airway contraction and Ca(2+) signalling, recorded via confocal microscopy, were induced with ACh, either alone or after pretreatment of slices with Zo.Cr and (or) verapamil, a standard Ca(2+) channel blocker. ACh (10 micromol/L) stimulated airway contraction, seen as decreased airway diameter, and also stimulated Ca(2+) transients (sharp rise in [Ca(2+)]i) and oscillations in ASMCs, seen as increased fluo-4-induced fluorescence intensity. When Zo.Cr (0.3-1.0 mg/mL) was given 30 min before ACh administration, the ACh-induced airway contraction and Ca(2+) signalling were significantly reduced. Similarly, verapamil (1 micromol/L) also inhibited agonist-induced airway contraction and Ca(2+) signalling, indicating a similarity in the modes of action. When Zo.Cr (0.3 mg/mL) and verapamil (1 micromol/L) were given together before ACh, the degree of inhibition was the same as that observed when each of these blockers was given alone, indicating absence of any additional inhibitory mechanism in the extract. In Ca(2+) -free solution, both Zo.Cr and verapamil, when given separately, inhibited Ca(2+) (10 mmol/L) induced increase in fluorescence and airway contraction. This shows that ginger inhibits airway contraction and associated Ca(2+) signalling, possibly via blockade of plasma membrane Ca(2+) channels, thus reiterating the effectiveness of this age-old herb in treating respiratory illnesses. PMID- 18432288 TI - Comparing the lactate and EMG thresholds of recreational cyclists during incremental pedaling exercise. AB - The purpose of this study was to determine the validity of using the electromyography (EMG) signal as a noninvasive method of estimating the lactate threshold (LT) power output in recreational cyclists. Using an electromagnetic bicycle ergometer and constant pedaling cadence of 80 rpm, 24 recreational cyclists performed an incremental exercise protocol that consisted of stepwise increases in power output of 25 W every 3 min until exhaustion. The EMG signal was recorded from the right vastus lateralis (VL) and right rectus femoris (RF) throughout the test. Blood samples were taken from the fingertip every 3 min. The LT was determined by examining the relation between the lactate concentration and the power output using a log-log transformation model. The root mean square (RMS) value from the EMG signal was calculated for every 1-second non-superimposing window. Sets of pairs of straight regression lines were plotted and the corresponding determination coefficients (R(2)) were calculated. The intersection point of the pair of lines with the highest R(2) product was chosen to represent the EMG threshold (EMGT). The results showed that the correlation coefficients (r) between EMGT and LT were significant (p < 0.01) and high for the VL (r = 0.826) and RF (r = 0.872). The RF and VL muscles showed similar behavior during the maximal incremental test and the EMGT and LT power output were equivalent for both muscles. The validity of using EMG to estimate the LT power output in recreational cyclists was confirmed. PMID- 18432289 TI - Potential cytoprotection: antioxidant defence by caffeic acid phenethyl ester against free radical-induced damage of lipids, DNA, and proteins. AB - Oxidative stress is considered to be a major cause of cellular injuries in a variety of chronic health problems, such as carcinogenesis and neurodegenerative disorders. Caffeic acid phenethyl ester (CAPE), derived from the propolis of honeybee hives, possesses a variety of biological and pharmacological properties including antioxidant and anticancer activity. In the present study, we focused on the diverse antioxidative functionalities of CAPE and its related polyphenolic acid esters on cellular macromolecules in vitro. The effects on human erythrocyte membrane ghost lipid peroxidation, plasmid pBR322 DNA, and protein damage initiated by the water-soluble initiator 2,2'-azobis(2-amidinopropane) hydrochloride (AAPH) and hydrogen peroxide (H(2)O(2)) were monitored by formation of hydroperoxides and by DNA nicking assay, single-cell alkaline electrophoresis, and SDS-polyacrylamide gel electrophoresis. Our results showed that CAPE and its related polyphenolic acid esters elicited remarkable inhibitory effects on erythrocyte membrane lipid peroxidation, cellular DNA strand breakage, and protein fragmentation. The results suggest that CAPE is a potent exogenous cytoprotective and antigenotoxic agent against cell oxidative damage that could be used as a template for designing novel drugs to combat diseases induced by oxidative stress components, such as various types of cancer. PMID- 18432290 TI - Pulmonary gemcitabine delivery for treating lung cancer: pharmacokinetics and acute lung injury aspects in animals. AB - Gemcitabine, a nucleoside analogue for treating lung cancer, is clinically administered as an intravenous infusion. To achieve better patient compliance and more direct effect on the lung, we explored a new gemcitabine pulmonary delivery route and evaluated the pharmacokinetics and acute lung injury aspects in animals. Pharmacokinetics of gemcitabine were measured in Sprague-Dawley rats after intravenous (i.v.), intratracheal instillation by tracheotomy (i.t.t.), intratracheal instillation via orotrachea (i.t.o.), and intragastric (i.g.) administration of gemcitabine. Acute lung injury effects of the pulmonary delivery of gemcitabine were performed in Sprague-Dawley rats after i.t.o. and i.v. administration of gemcitabine and i.t.o. administration of lipopolysaccharide (LPS) as a positive control and physiological saline as a blank control. Indicators for acute lung injury that were evaluated included lung morphology, lung histopathology, lung coefficient, lung wet/dry weight ratio, total cell and classification counts in bronchoalveolar lavage cells (BALC), and total protein and TNF-alpha levels in bronchoalveolar lavage fluids (BALF). After i.t.t. or i.t.o. administration, gemcitabine was quickly absorbed, but i.g. administration led to an undetectable plasma gemcitabine concentration. Absolute bioavailability of gemcitabine after i.t.t. and i.t.o. administration was 91% and 65%, respectively. Gemcitabine given as i.t.o. administration did not cause any overt acute lung injury. All indicators for acute lung injury in the i.t.o. group were similar to those in the i.v. group or in the blank control, but significantly different from those in the positive control. In conclusion, the pharmacokinetics and acute lung injury studies suggest that pulmonary gemcitabine delivery would be a new and promising administration route. PMID- 18432296 TI - Warning on hepatotoxicity of darunavir. PMID- 18432304 TI - Cytoplasmic maturation of mammalian oocytes: development of a mechanism responsible for sperm-induced Ca2+ oscillations. AB - The oocytes of most mammalian species, including mouse and human, are fertilized in metaphase of the second meiotic division. A fertilizing spermatozoon introduces an oocyte-activating factor, phospholipase C zeta, triggering oscillations of the cytoplasmic concentration of free calcium ions ([Ca(2+)](i)) in the oocyte. [Ca(2+)](i) oscillations are essential for the activation of the embryonic development. They trigger processes such as resumption and completion of meiosis, establishment of the block to polyspermy and recruitment of maternal mRNAs necessary for the activation of the embryo genome. Moreover, it has been recently shown that [Ca(2+)](i) oscillations may also influence the development of the embryo. The ability to generate [Ca(2+)](i) oscillations develops in mammalian oocytes during meiotic maturation and requires several cytoplasmic changes, including: 1/ reorganization of endoplasmic reticulum, the main stockpile of calcium in the oocyte, 2/ increase in the number of 1,4,5-inositol triphosphate (IP(3)) receptors, 3/ changes in their biochemical properties (e.g.: sensitivity to IP3), and possibly both 4/ an increase in the concentration of Ca(2+) ions stored in endoplasmic reticulum (ER) and 5/ redistribution of Ca(2+) binding ER proteins. The aim of this review is to present the state of current knowledge about these processes. PMID- 18432305 TI - Timing of the first zygotic cleavage as a marker of developmental potential of mammalian embryos. AB - Embryo quality related to its developmental potential is now one of the most important issues in modern embryology. It has been demonstrated that some in vitro produced blastocysts fail to hatch and implant after transfer despite a normal morphology. Although embryos are able to adjust to sub-optimal culture conditions, significant changes in expression profiles of developmentally important genes have been noticed. Timing of the first zygotic cleavage is considered a non-invasive marker of embryo developmental potential and has been successfully used in human IVF programs for identifying embryos of superior quality. Early-cleaving zygotes are more likely to develop to the blastocyst stage than their late-cleaving counterparts. The timing of the first zygotic cleavage has been associated with several parameters that may affect developmental potential of the resulting embryos. The mechanism causing variation in the timing of the first zygotic cleavage has not been identified. It may be related to culture environment or to some intrinsic factors within the oocyte, the sperm or both. In this paper we discuss some of the important aspects related to the timing of the first zygotic cleavage and its influence on the developmental competence of resulting embryos. PMID- 18432306 TI - Influence of estradiol-17beta and progesterone on nitric oxide (NO) production in the porcine endometrium during first half of pregnancy. AB - The purpose of the study was to examine: 1/ endometrial concentrations of nitrate/nitrite (NOx) in pregnant pigs, and 2/ the influence of estradiol-17beta (E(2)) and/or progesterone (P(4)) on NOx production by porcine endometrium during the first half of pregnancy. Total NOx concentrations were determined using a microplate assay method based on the Griess reaction. Evident fluctuations of endometrial NOx content were found during the examined time of pregnancy (days 5, 10, 15, 20, 25, 30, 35, 40 and 60 of pregnancy). The NOx concentration was highest on days 10 and 15, and then lowered until day 60 of pregnancy. In addition, we demonstrated the stimulatory effect of E(2) and/or P(4) on NO in vitro production by porcine endometrial slices. The medium content of NOx depended on the steroid type, treatment dose and day of pregnancy. It is possible that the observed differences in the strength of the stimulatory action of E(2) and/or P(4) on endometrial NOx production are associated with activation of different isoforms of NOS. PMID- 18432307 TI - The effect of tumor necrosis factor-alpha (TNF-alpha), interleukin (IL)-1 beta and IL-6 on chorioamnion secretion of prostaglandins (PG)F 2 alpha and E2 in pigs. AB - The aim of the present study was to determine the effect of tumor necrosis factor alpha (TNF-alpha), interleukin-1 beta (IL-1 beta) and interleukin-6 (IL-6) on prostaglandin (PG)F(2 alpha) and PGE(2) secretion as well as cyclooxygenase-2 (COX-2) protein expression in chorioamnion collected on days 25, 30 and 40 of pregnancy in pigs. Fetal membrane slices were incubated for 16 h with TNF-alpha, IL-1 beta, IL-6 (1 or 10 ng/ml of medium) or two combinations of the three cytokines (1 or 10 ng/ml of each cytokine per combination). We demonstrated the stimulatory effect of TNF-alpha, IL-1 beta and/or IL-6 on PGF(2 alpha) and PGE(2) secretion by the porcine fetal membranes. The medium content of these PGs depended on the cytokine type, treatment dose and day of pregnancy. Cytokine stimulation of PGE(2) was more pronounced than that of PGF(2 alpha). In addition, an increase in PGF(2 alpha) and/or PGE(2) secretion was usually associated with an augmentation of COX-2 protein expression. Our results support the notion concerning the possible role of cytokines in modulating production of PGs by fetal membranes during the first trimester of gestation. PMID- 18432308 TI - Maximum rates of cooling by three programmable freezers, and the potential relevance to sperm cryopreservation. AB - Maximum rates of cooling for the Asymptote EF 100 and the Cryologic CL8800 temperature controller with either a standard or fast chamber were determined and viewed in the context of sperm cryopreservation. All three systems use liquid nitrogen to cool the plate or chamber which would hold the sample, opposed by a variable amount of heat from an internal heater. Maximum rates of cooling for all systems were a function of the temperature gradient between the liquid nitrogen and the plate/chamber and at a plate/chamber temperature of 15 degrees C were 16.5 degrees C/min, 13.3 degrees C/min and 8.0 degrees C/min for the Asymptote EF100, Cryologic fast and slow chambers respectively. These machines are not suited to the freezing of sperm from species requiring rapid rates of cooling, an important consideration when planning to purchase a piece of equipment for this application, and scientists are advised to discuss specifically their requirements with prospective suppliers. PMID- 18432310 TI - Vgf is a novel biomarker associated with muscle weakness in amyotrophic lateral sclerosis (ALS), with a potential role in disease pathogenesis. AB - Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disease that affects nerve cells in the brain and the spinal cord. Previous proteomic evidence revealed that the content of certain peptide fragments including Vgf derived peptide aa 398-411 (Vgf(398-411)) of the precursor Vgf protein in the cerebral spinal fluid (CSF) correctly identified patients with ALS from normal and disease controls. Using quantitative ELISA immunoassay we found that the CSF levels of Vgf decreases with muscle weakness in patients with ALS. In SOD1 G93A transgenic mice, loss of full-length Vgf content in CSF, serum and in SMI-32 immunopositive spinal cord motor neurons is noted in asymptomatic animals (approximately 75 days old) and continues to show a progressive decline as animals weaken. In vitro studies show that viral-mediated exogenous Vgf expression in primary mixed spinal cord neuron cultures attenuates excitotoxic injury. Thus, while Vgf may be a reliable biomarker of progression of muscle weakness in patients with ALS, restoration of Vgf expression in spinal cord motor neurons may therapeutically rescue spinal cord motorneurons against excitotoxic injury. PMID- 18432311 TI - Altered Phase Interactions between Spontaneous Blood Pressure and Flow Fluctuations in Type 2 Diabetes Mellitus: Nonlinear Assessment of Cerebral Autoregulation. AB - Cerebral autoregulation (CA) is an important mechanism that involves dilation and constriction in arterioles to maintain relatively s cerebral blood flow in response to changes of systemic blood pressure. Traditional assessments of CA focus on the changes of cerebral blood flow velocity in response to large blood pressure fluctuations induced by interventions. This approach is not feasible for patients with impaired autoregulation or cardiovascular regulation. Here we propose a newly developed technique-the multimodal pressure-flow (MMPF) analysis, which assesses CA by quantifying nonlinear phase interactions between spontaneous oscillations in blood pressure and flow velocity during resting conditions. We show that CA in healthy subjects can be characterized by specific phase shifts between spontaneous blood pressure and flow velocity oscillations, and the phase shifts are significantly reduced in diabetic subjects. Smaller phase shifts between oscillations in the two variables indicate more passive dependence of blood flow velocity on blood pressure, thus suggesting impaired cerebral autoregulation. Moreover, the reduction of the phase shifts in diabetes is observed not only in previously-recognized effective region of CA (<0.1Hz), but also over the higher frequency range from ~0.1 to 0.4Hz. These findings indicate that Type 2 diabetes alters cerebral blood flow regulation over a wide frequency range and that this alteration can be reliably assessed from spontaneous oscillations in blood pressure and blood flow velocity during resting conditions. We also show that the MMPF method has better performance than traditional approaches based on Fourier transform, and is more sui for the quantification of nonlinear phase interactions between nonstationary biological signals such as blood pressure and blood flow. PMID- 18432312 TI - Regulatory function of the P295-T311 motif of the estrogen receptor alpha - does proteasomal degradation of the receptor induce emergence of peptides implicated in estrogenic responses? AB - The way in which estrogen receptor alpha (ERalpha) mediates gene transcription and hormone-dependent cancer cell proliferation is now being largely reconsidered in view of several recent discoveries. ERalpha-mediated transcription appears to be a cyclic and transient process where the proteasome - and thus receptor degradation - plays a pivotal role. In view of our recent investigations, which demonstrate the estrogenic activity of a synthetic peptide corresponding to a regulatory motif of the receptor (ERalpha17p), we propose that ERalpha proteasomal degradation could induce the emergence of regulatory peptide(s). The latter would function as a signal and contribute to the ERalpha activation process, amplifying the initial hormonal stimulation and giving rise to sustained estrogenic response. PMID- 18432313 TI - E6-associated protein (E6-AP) is a dual function coactivator of steroid hormone receptors. AB - Steroid hormone receptors (SHR) belong to a large family of ligand-activated transcription factors that perform their biological functions by enhancing the transcription of specific target genes. The transactivation functions of SHRs are regulated by a specialized group of proteins called coactivators. The SHR coactivators represent a growing class of proteins with various enzymatic activities that serve to modify the chromatin to facilitate the transcription of SHR target genes. The ubiquitin-proteasome pathway enzymes have also been added to the growing list of enzymatic activities that are recruited to the SHR target gene promoters during transcription. One such ubiquitin-proteasome pathway enzyme to be identified and characterized as a SHR coactivator was E6-associated protein (E6-AP). E6-AP is a hect (homologous to E6-associated protein carboxy-terminal domain) domain containing E3 ubiquitin ligase that possesses two independent separable functions; a coactivation function and an ubiquitin-protein ligase activity. Being a component of the ubiquitin-proteasome pathway, it is postulated that E6-AP may orchestrate the dynamics of steroid hormone receptor-mediated transcription by regulating the degradation of the transcriptional complexes. E6 AP has also been shown to be involved in the regulation of various aspects of reproduction such as prostate and mammary gland development. Furthermore, it has been demonstrated that E6-AP expression is down-regulated in breast and prostate tumors and that the expression of E6-AP is inversely associated with that of estrogen and androgen receptors. This review summarizes our current knowledge about the structures, molecular mechanisms, spatiotemporal expression patterns and biological functions of E6-AP. PMID- 18432314 TI - A novel splice-site mutation of TULP1 underlies severe early-onset retinitis pigmentosa in a consanguineous Israeli Muslim Arab family. AB - PURPOSE: To investigate the genetic basis for autosomal recessive severe early onset retinitis pigmentosa (RP) in a consanguineous Israeli Muslim Arab family. METHODS: Haplotype analysis for all known genes underlying autosomal recessive RP was performed. Mutation screening of the underlying gene was done by direct sequencing. An in vitro splicing assay was used to evaluate the effect of the identified mutation on splicing. RESULTS: Haplotype analysis indicated linkage to the Tubby-like protein 1 (TULP)1 gene. Direct sequencing revealed a homozygous single base insertion, c.1495+2_1495+3insT, located in the conserved donor splice site of intron 14. This mutation co-segregated with the disease, and was not detected in 114 unrelated Israeli Muslim Arab controls. We used an in vitro splicing assay to demonstrate that this mutation leads to incorrect splicing. CONCLUSIONS: To date, 22 distinct pathogenic mutations of TULP1 have been reported in patients with early-onset RP or Leber congenital amaurosis. Here we report a novel splice-site mutation of TULP1, c.1495+2_1495+3insT, underlying autosomal recessive early-onset RP in a consanguineous Israeli Muslim Arab family. This report expands the spectrum of pathogenic mutations of the TULP1 gene. PMID- 18432315 TI - Cone and rod cells have different target preferences in vitro as revealed by optical tweezers. AB - PURPOSE: When neural circuits are damaged in adulthood, regenerating and sprouting processes must distinguish appropriate targets to recreate the normal circuitry. We tested the ability of adult nerve cells to target specific cells in culture using the retina as a model system. METHODS: Under sterile culture conditions, retinal cells, isolated from tiger salamander retina, were micromanipulated with optical tweezers to create pairs of first-order photoreceptor cells with second- or third-order retinal neurons. The development of cell contact and presynaptic varicosities, the direction and amount of neuritic growth, and nerve cell polarity were assessed after seven days in vitro. Cultures were labeled for rod opsin to distinguish rod from cone cells and for the alpha subunit of the trimeric G protein Go (Go alpha) to identify cone dominated and mixed rod-cone ON bipolar cells. RESULTS: Quantitative analysis of growth demonstrated that target preferences were cell-specific: Cone cells preferred second-order bipolar cells, whereas rod cells grew toward third-order neurons, which include amacrine and ganglion cells. In addition, when rod cells grew toward bipolar cells, they chose an abnormally high number of Go alpha positive bipolar cells. These growth patterns were not affected by tweezers manipulation or the amount of growth. Cell orientation of the photoreceptor also did not affect preferences: Cells oriented away from dendritic processes could reorient their axonal pole toward the target cell. CONCLUSIONS: Cone cells preferred normal partners, and rod cells preferred novel partners. These intrinsic preferences indicate that adult nerve cells can have differing capacities for targeting even if they come from the same cell class. Further,these differences may help explain the patterns of photoreceptor sprouting seen in retinal degeneration in which rod, but not cone, cells invade the inner retinal layers where third-order neurons are located. PMID- 18432316 TI - A novel nonsense mutation in CRYBB1 associated with autosomal dominant congenital cataract. AB - PURPOSE: To identify the molecular defect underlying an autosomal dominant congenital nuclear cataract in a Chinese family. METHODS: Twenty-two members of a three-generation pedigree were recruited, clinical examinations were performed, and genomic DNA was extracted from peripheral blood leukocytes. All members were genotyped with polymorphic microsatellite markers adjacent to each of the known cataract-related genes. Linkage analysis was performed after genotyping. Candidate genes were screened for mutation using direct sequencing. Individuals were screened for presence of a mutation by restriction fragment length polymorphism (RFLP) analysis. RESULTS: Linkage analysis identified a maximum LOD score of 3.31 (recombination fraction [theta]=0.0) with marker D22S1167 on chromosome 22, which flanks the beta-crystallin gene cluster (CRYBB3, CRYBB2, CRYBB1, and CRYBA4). Sequencing the coding regions and the flanking intronic sequences of these four candidate genes identified a novel, heterozygous C-->T transition in exon 6 of CRYBB1 in the affected individuals of the family. This single nucleotide change introduced a novel BfaI site and was predicted to result in a nonsense mutation at codon 223 that changed a phylogenetically conserved amino acid to a stop codon (p.Q223X). RFLP analysis confirmed that this mutation co-segregated with the disease phenotype in all available family members and was not found in 100 normal unrelated individuals from the same ethnic background. CONCLUSIONS: This study has identified a novel nonsense mutation in CRYBB1 (p.Q223X) associated with autosomal dominant congenital nuclear cataract. PMID- 18432317 TI - A genome-wide scan maps a novel autosomal dominant juvenile-onset open-angle glaucoma locus to 2p15-16. AB - PURPOSE: To study the clinical features and to perform genetic linkage study in two large Chinese families with autosomal dominant juvenile-onset primary open angle glaucoma (POAG). METHODS: Eighteen members of one Chinese family and 25 members of a second Chinese family with juvenile-onset primary open-angle glaucoma (POAG) were investigated. Thirteen members in one family and 14 members in the second family were diagnosed with juvenile-onset POAG. A genome-wide linkage scan was performed on one family using 411 short tandem repeat (STR) markers. Subsequent fine mapping was performed in the two study families using a modified fluorescent labeled M13 primer method. RESULTS: A whole genome-wide scan in one family showed linkage to chromosome 2p15-p16 with a two-point maximum LOD score of 5.01 at theta=0 between the disease phenotype and STR marker D2S337. The second family was also mapped to the same locus with a two-point maximum LOD score of 6.30 at theta=0 for D2S378. Haplotype analysis in these two families demonstrated that they shared the same disease haplotype, suggesting they have inherited the mutation from a common founder. The maximum LOD scores were 8.93 at theta=0 for D2S378 and 9.9 at theta=0 for D2S337 when the two families were combined for analysis. The disease interval for these two families was localized to 9.2 cM or 13.3 Mb between D2S123 and D2S2397. There are 42 known genes/transcripts within the interval. Five of these genes were sequenced, and no disease-causing mutation was identified in either family. CONCLUSIONS: This novel juvenile-onset POAG locus on chromosome 2p15-16 is overlapped by the Glaucoma 1, open angle, H (GLC1H) locus for adult-onset POAG. Eventual identification of the disease-causing gene will provide insights into the pathogenesis of POAG. PMID- 18432318 TI - Differential expression of the catalytic subunits for PP-1 and PP-2A and the regulatory subunits for PP-2A in mouse eye. AB - PURPOSE: Reversible protein phosphorylation is a fundamental regulatory mechanism in all biologic processes. Protein serine/threonine phosphatases-1 (PP-1) and 2A (PP-2A) account for 90% of serine/threonine phosphatase activity in eukaryote cells and play distinct roles in regulating multiple cellular processes and activities. Our previous studies have established the expression patterns of the catalytic subunits for PP-1 (PP-1cs) and PP-2A (PP-2Acs) in bovine and rat lenses. In the present study, we have determined the expression patterns of PP 1cs (PP-1alpha and PP-1beta) and PP-2Acs (PP-2Aalpha and PP-2Abeta) in the retina and cornea along with the ocular lens of the mouse eye. Moreover, since the function of PP-2A is largely relied on its regulatory subunits, we have also analyzed the expression patterns of the genes encoding the scaffold A subunits of PP-2A, PP2A-Aalpha and PP2A-Abeta, and the regulatory B family subunits of PP-2A, PP2A-Balpha, PP2A-Bbeta, and PP2A-Bgamma. In addition, we have also demonstrated the differential protections of PP-1 and PP-2A in mouse lens epithelial cell line, alphaTN4-1, against oxidative stress-induced apoptosis. METHODS: Total RNAs and proteins were extracted from the retina, lens epithelium, lens fiber cells, and cornea of the mouse eye. Reverse transcription polymerase chain reaction (RT PCR) and real time PCR were used to detect the mRNA expression. Western blot and immunohistochemistry analysis were applied to examine the protein expression and distribution. Stable clones of alphaTN4-1 cells expressing either PP-1alpha or PP 2Aalpha were used to analyze the differential protections against oxidative stress-induced apoptosis. RESULTS: PP-1 is more abundant than PP-2A in the mouse eye. The catalytic subunits for PP-1 and PP-2A display similar expression patterns in the retina and cornea but much reduced in the lens. The mRNAs for all five isoforms of PP2A-A and PP2A-B subunits are highly expressed in the retina, but only three out of the five mRNAs are expressed in the cornea. In the ocular lens, only PP2A-Abeta and PP2A-Bgamma mRNAs are clearly detectable. The A and B subunit proteins of PP-2A are highly expressed in the retina and cornea but are much reduced in the ocular lens. PP2A-Aalpha/beta are differentially distributed in the mouse retina.When transfected into mouse lens epithelial cells, alphaTN4 1, PP-1alpha and PP-2Aalpha display differential protection against oxidative stress-induced apoptosis. CONCLUSIONS: Our results lead to the following conclusions regarding PP-1 and PP-2A in mouse eye: 1) PP1 is a more abundant phosphatase than PP-2A; 2) both PP-1 and PP-2A may play important roles, and the functions of PP-2A appear to be highly regulated by various regulatory subunits; and 3) the genes encoding PP-1alpha/beta, PP-2Aalpha/beta, PP-2A-Aalpha/beta, and PP-2A-B alpha/beta/gamma are all differentially expressed. PMID- 18432319 TI - [Septic arthritis: what is the role for the rheumatologist?]. AB - Septic arthritis (SA) is a clinical emergency with considerable morbidity and mortality that can lead to rapid joint destruction and irreversible loss of function. The reported incidence varies from 2-5 cases per 100.000 individuals per year in the general populations to 70 cases per 100.000 individuals annually among patients with rheumatoid arthritis (RA). Predisposing factors are immunosuppressive and corticosteroids therapy and RA "itself". The expected decrease in incidence of SA was not seen over the last 20 years period but we can, on the contrary, expect an increase in the frequency of its appearance because of the population ageing, the increasingly prosthetic joint replacement, the ability of the bacteria to evade clearance by the host immune response and the rapidly growing number of patients with RA, ankylosing spondylitis and psoriatic arthritis treated with tumour necrosis factor alpha (TNF alpha) antagonists. Up to now there have been conflicting reports regarding joint infections in patients under anti-TNF therapy but according to data from Deutsch as well as the British register there might be an increase in the incidence of joint infections in anti-TNF treated patients. Microscopic analysis and culture of synovial fluid are fundamental diagnostic tools in the evaluation of possible joint sepsis. Sonographic guidance of arthrocentesis led to successful aspiration of difficult-to-access joints as shoulder and hip. There is controversy over which mode of drainage of septic synovial fluid should be employed but needle aspiration appear to be preferable to surgical treatment as an initial mode of treatment of SA. Rheumatologists should have a central role in the diagnosis and management of SA. PMID- 18432320 TI - [Bisphosphonate-associated osteonecrosis of the jaw in rheumatology: a systematic review]. AB - Osteonecrosis of the jaw (ONJ) is a well-known devastating side effect of parenteral bisphosphonate therapy for cancer. Several ONJ cases have been reported in patients taking oral bisphosphonates for osteoporosis or Paget's disease. Even if the number of cases of ONJ in patients taking oral bisphosphonates are still rare compared to the total exposure, rheumatologists treating bone diseases with bisphosphonates must be aware of this new complication, allowing for prevention and early diagnosis. The patients must be informed on the benefit/risk of bisphosphonate therapy and, when necessary and possible, alternative therapy for postmenopausal osteoporosis should be considered. The need for the patient to be dentally fit and to maintain this state forever should be part of the informed consent for bisphosphonate treatment. It is uncommon for rheumatologists to ask about dental problems but this new bisphosphonate- associated complication highlights the need for this to change. In this paper we review the literature available on this newly described bisphosphonate-induced complication with particular emphasis on ONJ cases related to the use of oral bisphosphonates. PMID- 18432321 TI - [Adult Scheuermann's disease as cause of mechanic dorsalgia]. AB - Scheuermann's disease (SD) or vertebral osteochondrosis is the most frequent cause of non postural kyphosis and one of more frequent cause of adolescent's dorsalgia. The criteria for the diagnosis are: more than 5 degrees of wedging of at least three adjacent vertebrae at the apex of the kyphosis; a toracic kyphosis of more than 45 of Cobb's degree; Schmorl's nodes and endplates irregularities. In addition to classic SD, there are radiological alterations that remain asymptomatic for a long time to reveal in adult age: in that case it speaks of adult Scheuermann's disease (ASD). We considered the diagnosis of patients came from April 2006 to April 2007 on Day Hospital in our Clinic. ASD was diagnosed, besides, in 10 of these patients. 7 patients had previous diagnosis such as: dorsal Spondiloarthrosis (4 subjects); Osteoporosis with vertebral fractures (3 subjects). All these diagnosis was not confirmed by us. In case of chronic dorsalgia of adult, ASD is rarely considered as differential diagnosis. Besides, the vertebral dorsalgia, even in absence of red flags as fever, asthenia,hypersedimetry, functional loss and aching spinal processes to tapping, could hide a serious scene that lead us to be careful in the differential diagnosis, because of similar radiological pictures of the MSA to other pathology as spondylodiscitis, primitive or metastasic spinal tumors, and brittleness vertebral fractures. PMID- 18432323 TI - [Analysis of the evolution to defined connective tissue diseases of patients with "early unidifferentiated connective tissue diseases (UCTD)"]. AB - The term undifferentiated connective tissue diseases (UCTD) is used to identify systemic autoimmune diseases not fulfilling classificative criteria for defined connective tissue diseases (CTD). Aim of the present study was to evaluate the evolution to defined CTD of an historical cohort of 91 UCTD patients followed at our Unit and to describe clinical and serological characteristics of stable UCTD patients with a disease duration of more than 5 years. Patients, previously described, were selected for having an undifferentiated profile after 1 year of follow up. These patients have been regularly followed at our Unit and their diagnosis has been reassessed annually based on the existing classificative criteria. Seven UCTD patients with a follow up of less than 5 years have been excluded from the study, therefore 84 patients (F: 81, M: 3) have been analysed. During the follow up 28 patients (33%) developed a defined CTD. In particular 22 patients developed systemic lupus erythematosus (SLE), while the remaining 6 patients developed other CTDs (2 primary Sjogren's syndrome, 2 overlap syndromes, 1 Systemic Sclerosis, 1 rheumatoid arthritis). The evolution to a defined CTD occurred after a mean disease duration of 80.6+/- 66.8 months (min 14, max 336, median 72); the evolution to SLE occurred after a mean disease duration of 66.8+/ 43.3 months (min 17, max 216, median 57). Anti-cardiolipin antibodies were the only variable correlated with the evolution to SLE (p<0.05). Stable UCTD were characterized by a simplified clinical picture with no major organ involvement and by a simplified autoantibody profile (anti-Ro/SSA antibodies and anti-RNP antibodies were the single antibody specificities observed in 22% and 13% of patients respectively). These results confirm previous data showing that about 30% of UCTD patients will develop a defined CTD, the predictive role of anti cardiolipin antibodies for the evolution to SLE, and the existence of stable UCTD, distinct clinical entities with a simplified clinico-serological profile. The early identification of stable UCTD is very important both from a clinical and a research point of view. Future research is needed to define a new set of classification criteria. PMID- 18432322 TI - Prophylaxis and therapy of HBV infection in 20 patients treated with disease modifying antirheumatic drugs or with biological agents for rheumatic diseases. AB - OBJECT: To evaluate the safety and tolerability of lamivudine in patients with HBV infection needing immunosuppressive treatment for rheumatic diseases. PATIENTS AND METHODS: Twenty patients with rheumatic diseases planned to receive immunosuppressive DMARDs or biological agents were screened for HBV markers. In all active carriers antiviral treatment was recommended. Inactive carriers (HBsAg positive, aminotrasferase and viremia persistently normal) were divided into two risk categories according to the type and the degree of immunosuppression, and antiviral prophylaxis was started only in patients of the high risk category. Antiviral treatment was recommended also in potential occult carriers (HBsAg negative, HBcAb positive) treated with rituximab. In twenty patients antiviral treatment was started: 1 was a potential occult carrier planned to receive rituximab; 9 were inactive carriers, in which prophylactic therapy was needed for a high risk of HBV reactivation (in 3, for the use of TNF blocking agents); 10 were treated for active viral replication. Prophylaxis and therapy were performed with lamivudine. In three patients adefovir was associated. RESULTS: Antiviral drugs were well tolerated. In all cases, immunosuppressive treatment was given for the planned duration of therapy, with good results on the rheumatic diseases. Median duration of antiviral treatment was 19 months (for a total of 386 month/person). No cases of viral reactivation were observed. CONCLUSION: Our experience demonstrates the feasibility of a prophylaxis and therapy of HBV infection in patients with rheumatic diseases. This approach reduces the risk of viral reactivation and allows the choice of the optimal immunosuppressive treatment in rheumatic patients. PMID- 18432324 TI - [Anti-TNFalpha agents in elderly patients with rheumatoid arthritis: a study of a group of 105 over sixty five years old patients]. AB - OBJECTIVE: To evaluate the efficacy and the safety of anti-TNF alfa treatment in elderly patients (>/=65 years old) with active rheumatoid arthritis (RA), in comparison with younger (17-65 years old). METHODS: We considered retrospectively 295 patients, affected by RA and treated with anti-TNF alfa drugs. They were divided in two groups, according to their age, and followed up for two years: over-65-years old patients (190) and under-65-years old patients (105). Effectiveness of drugs was assessed analyzing RA disease activity (DAS28, DAS44, SDAI), functional status (HAQ) and serological parameters (CRP) before and after anti-TNF alfa therapy. Safety was studied considering discontinuation rate of biological disease-modifying antirheumatic drugs, and collateral events rate. RESULTS: At baseline, elderly patients showed higher disease activity's score (DAS 28, DAS44, SDAI, HAQ) with important loss of articular function (worse quality of life, HAQ) than younger patients (p<0.05). During the therapy, improvement in clinical parameters was observed (DAS28, DAS44 and SDAI) with no significant difference between the two groups. In elderly patients disability index, on the contrary, improved less than in younger (p<0.05). After treatment, also CRP decreased less in elderly patients (p<0.05). During the follow-up, 74 over-65-years old patients (38.95%) and 116 under-65-years old patients (38.05%) discontinued anti-TNF alfa therapy because of loss of efficacy (20.52% vs 11.42%), severe adverse events (17.34% vs 25.67%), voluntary discontinuation or good clinical response (1% vs 0.95%). No differences were shown about the frequency and reasons of anti-TNF alfa withdrawal (p>0.05). CONCLUSIONS: Anti-TNF alfa treatment was efficacious and safe in both groups of patients. These drugs induced improvement in disease activity, apart from the age. No functional improvement was observed in HAQ, showing the irreversible loss of articular function and the incomplete recovery in elderly patients. Age doesn't interfere with the possibility to treat elderly patients with anti-TNF alfa drugs. PMID- 18432325 TI - [Alexithymia and immunoendocrine parameters in patients affected by systemic lupus erythematosus and rheumatoid arthritis]. AB - OBJECTIVE: Aim of this study was to evaluate the prevalence of alexithymia in patients affected by SLE or RA and to investigate the correlation between alexithymia and immunoendocrine parameters (PRL, hGH, IL-6 and TNF-alfa). METHODS: Twenty-five patients (12 and 13 affected by SLE and RA, respectively) were enrolled into the study. The Toronto Alexithymia Scale-20 (TAS-20) was administered. PRL, hGH, IL-6 and TNF-alfa levels were measured by commercially available ELISA kits. RESULTS: Alexithymia prevalence (TAS-20 > or = 51) was 54% in RA and 42% in SLE patients. hGH serum levels were 3.1+/-4.2 and 1.1+/-0.9 IU/ml in SLE and RA, respectively. PRL concentration was 18.4+/-6.5 ng/ml and 14.2+/-4.0 ng/ml in SLE and RA patients, respectively (p=0.03). In RA group, TNF alpha was 20+/-36.2 whereas in SLE it was 4.9+/-12.8 pg/ml (p=0.03); IL-6 serum concentrations were 24.4+/-25.1 and 2.9+/-5.4 pg/ml, in RA and SLE respectively (p=0.004). The serum level of hGH showed slight increase in alexithymic group (A) compared to non alexithymic group (NA) in both SLE and RA patients. PRL serum levels in SLE-A patients was 26.7+/-17.3 ng/ml while in SLE-NA patients was 12.4+/-3.3 ng/ml (p=0.04). In RA patients increased values of IL-6 and TNF-alpha were present in the A group compared to NA group (IL-6: 35.3+/-28 pg/mL vs 3.5+/ 3.9 pg/mL, p=0.01; TNF-alpha: 34.7+/-39 pg/mL vs 3.1+/-3.4 pg/mL, p=0.01). CONCLUSIONS: In this preliminary results we found an high prevalence of alexithymia and a correlation between immunoendocrine parameters and alexhytimic features in SLE and RA, suggesting that an immunomodulatory pathway could influence this cognitive style in patients with autoimmune disorders. Other studies should contribute to find a common biological pathway linking alexithymia and autoimmunity. PMID- 18432326 TI - [IFN-alpha-induced psoriatic arthritis and HCV-related liver cirrhosis. Therapeutic options and patient's opinion]. AB - Hepatitis C virus (HCV) infection in the setting of Psoriatic Arthritis is an additional variable to be considered in the therapeutic approach to the disease because of the complications of an immunosuppressive treatment in the course of a chronic infection and the possible hepatotoxicity of many drugs conventionally used to treat psoriatic arthritis. The case reported explores the therapeutic options in a patient with IFN-alpha-induced psoriatic arthritis, characterised by severe arthritis and psoriasis but also the concomitant presence of HCV chronic hepatitis, in light of the patient's concerns. PMID- 18432327 TI - [Neprhrotic syndrome in a patient with rheumatoid arthritis treated with adalimumab: a case report]. AB - The pathogenetic role of TNFalpha in inflammatory diseases has been known for a long time and has modified the therapeutic approach towards this pathology. All over the word, about 400.000 patients have been treated with biological anti-TNF alpha drugs. Particular attention has been taken for the safety of their use. We describe a case of 60 year old man affected by rheumatoid arthritis who has developed nephrotic syndrome with histological pattern typical of systemic lupus erythematosus-like syndrome, without autoantibodies. PMID- 18432328 TI - [Inscriptions of Sanationes from Sanctuaries of god Asclepius. Miraculous recoveries of rheuma diseases]. AB - Inscriptions of Sanationes from Sanctuaries of god Asclepius, testifying to the gratitude of patients to the divinity, present a situation, albeit fragmentary, of main diseases which physicians thought incurable or hardly curable. Sterility, calvities, worms, paralysis, as well as clinical disorders caused by rheuma appear, along with gout/podagra, in many inscriptions too. To counter them, patients asked for the intervention of Asclepius. PMID- 18432330 TI - Improving fluorescence detection in lab on chip devices. PMID- 18432331 TI - Sub-cellular precision on-chip small-animal immobilization, multi-photon imaging and femtosecond-laser manipulation. AB - Techniques for stable, rapid and repeatable small-animal immobilization are necessary for high-throughput in vivo genetic/drug screens using cellular and sub cellular features in multi-cellular organisms. We demonstrate a method for non invasive and high-throughput on-chip immobilization of physiologically active C. elegans without the use of anesthesia or cooling, but with comparable stability even for the most demanding purposes. We show observation and manipulation of sub cellular features in immobilized animals using two-photon microscopy and femtosecond-laser microsurgery. PMID- 18432332 TI - Tuneable separation in elastomeric microfluidics devices. AB - We describe how the elastomeric properties of PDMS (polydimethylsiloxane) can be utilised to achieve tuneable particle separation in Deterministic Lateral Displacement devices via strain controlled alteration of inter-obstacle distances, a development that opens up new avenues toward more effective separation of particles in microfluidics devices. PMID- 18432333 TI - A disposable planar peristaltic pump for lab-on-a-chip. AB - We demonstrate a simple planar peristaltic pump fabricated in poly(dimethylsiloxane) (PDMS) via soft lithography and suitable for microfluidic integration. PMID- 18432334 TI - Fabrication of a modular tissue construct in a microfluidic chip. AB - By combining microfluidics and soft-lithographic molding of gels containing mammalian cells, a device for three-dimensional (3D) culture of mammalian cells in microchannels was developed. Native components of the extracellular matrix, including collagen or Matrigel, made up the matrix of each molded piece (module) of cell-containing gel. Each module had at least one dimension below approximately 300 microm; in modules of these sizes, the flux of oxygen, nutrients, and metabolic products into and out of the modules was sufficient to allow cells in the modules to proliferate to densities comparable to those of native tissue (10(8)-10(9) cells cm(-3)). Packing modules loosely into microfluidic channels and chambers yielded structures permeated with a network of pores through which cell culture medium could flow to feed the encapsulated cells. The order in the packed assemblies increased as the width of the microchannels approached the width of the modules. Multiple cell types could be spatially organized in the small microfluidic channels. Recovery and analysis of modules after 24 h under constant flow of medium (200 microL h(-1)) showed that over 99% of encapsulated cells survived this interval in the microfluidic chamber. PMID- 18432335 TI - All-terrain droplet actuation. AB - Digital microfluidics has become a popular tool for biochemical and biomedical applications. However, its current format is restricted to actuation of droplets on a single plane. Here, we introduce a new method for fluid handling on flexible devices, which we have termed all-terrain droplet actuation (ATDA). We show that ATDA can be used to manipulate droplets across a wide range of geometries, including inclined, declined, vertical, twisted, and upside-down architectures. These new geometries enable flexible, straightforward integration of distinct physicochemical environments on monolithic devices. To illustrate this capacity, we developed temperature- and oxygen-sensitive colorimetric sensors, as well as an automated method for selective enrichment of DNA from a heterogeneous mixture. We anticipate that ATDA will be a useful new tool in the growing trend toward laboratory miniaturization. PMID- 18432336 TI - On a chip demonstration of a functional role for Odorant Binding Protein in the preservation of olfactory receptor activity at high odorant concentration. AB - The molecular mechanisms underlying odorant detection have been investigated using the chip based SPR technique by focusing on the dynamic interactions between transmembrane Olfactory Receptor OR1740, odorant ligands and soluble Odorant-Binding Protein (OBP-1F). The OR1740 present in the lipid bilayer of nanosomes derived from transformed yeasts specifically bound OBP-1F. The receptor preferential odorant ligand helional released bound OBP-1F from the OR-OBP complex, while unrelated odorants failed to do so. OBP-1F modified the functional OR1740 dose-response to helional, from a bell-shaped to a saturation curve, thus preserving OR activity at high ligand concentration. This unravels an active role for OBPs in olfaction, in addition to passive transport or a scavenger role. This sensorchip technology was applied to assessing native OBP-1F in a biological sample: rat olfactory mucus also displayed significant binding to OR1740 nanosomes, and the addition of helional yielded the dissociation of mucus OBP from the receptor. PMID- 18432337 TI - Lab-on-chip methodologies for the study of transport in porous media: energy applications. AB - We present a lab-on-chip approach to the study of multiphase transport in porous media. The applicability of microfluidics to biological and chemical analysis has motivated much development in lab-on-chip methodologies. Several of these methodologies are also well suited to the study of transport in porous media. We demonstrate the application of rapid prototyping of microfluidic networks with approximately 5000 channels, controllable wettability, and fluorescence-based analysis to the study of multiphase transport phenomena in porous media. The method is applied to measure the influence of wettability relative to network regularity, and to differentiate initial percolation patterns from active flow paths. Transport phenomena in porous media are of critical importance to many fields and particularly in many energy-related applications including liquid water transport in fuel cells, oil recovery, and CO(2) sequestration. PMID- 18432338 TI - Microfluidic device for immunoassays based on surface plasmon resonance imaging. AB - We have designed and fabricated a polydimethylsiloxane (PDMS) microfluidic device containing an array of gold spots onto which antigens or antibodies of interest can be attached. We use surface plasmon resonance (SPR) imaging to monitor the antibody-antigen recognition and binding events. This combination offers two significant advantages: (1) the microfluidic device dramatically reduces reaction time and sample consumption; and (2) the SPR imaging yields real-time detection of the immunocomplex formation. Thus, an immunoreaction may be detected and quantitatively characterized in about 10 min. The sensitivity of this method is at the subnanomolar level. When gold nanoparticles are selectively coupled to the immunocomplex to cause signal amplification, the sensitivity reaches the ten to one hundred picomolar level but the time required increases to about 60 min. PMID- 18432339 TI - A membrane-based microfluidic device for controlling the flux of platelet agonists into flowing blood. AB - The flux of platelet agonists into flowing blood is a critical event in thrombosis and hemostasis. However, few in vitro methods exist for examining and controlling the role of platelet agonists on clot formation and stability under hemodynamic conditions. In this paper, we describe a membrane-based method for introducing a solute into flowing blood at a defined flux. The device consisted of a track-etched polycarbonate membrane reversibly sealed between two microfluidic channels; one channel contained blood flowing at a physiologically relevant shear rate, and the other channel contained the agonist(s). An analytical model described the solute flux as a function of the membrane permeability and transmembrane pressure. The model was validated using luciferase as a model solute for transmembrane pressures of 50-400 Pa. As a proof-of concept, the weak platelet agonist ADP was introduced into whole blood flowing at 250 s(-1) at three fluxes (1.5, 2.4, and 4.4 x 10(-18) mol microm(-2) s(-1)). Platelet aggregation was monitored by fluorescence microscopy during the experiment and the morphology of aggregates was determined by post hoc confocal and electron microscopy. At the lowest flux (1.5 x 10(-18) mol microm(-2) s(-1)), we observed little to no aggregation. At the higher fluxes, we observed monolayer (2.4 x 10(-18) mol microm(-2) s(-1)) and multilayer (4.4 x 10(-18) mol microm(-2) s(-1)) aggregates of platelets and found that the platelet density within an aggregate increased with increasing ADP flux. We expect this device to be a useful tool in unraveling the role of platelet agonists on clot formation and stability. PMID- 18432341 TI - Automated cell culture in high density tubeless microfluidic device arrays. AB - Microfluidics is poised to have an impact on life sciences research. However, current microfluidic methods are not compatible with existing laboratory liquid dispensing and detection infrastructure. This incompatibility is a barrier to adoption of microfluidic systems and calls for improved approaches that will enhance performance and promote acceptance of microfluidic systems in the life sciences. Ease of use, standardized interfaces and automation remain critical challenges. We present a platform based on surface tension effects, where the difference in pressure inside drops of unequal volume drives flow in passive structures. We show integration with existing laboratory infrastructure, microfluidic operations such as pumping, routing and compartmentalization without discrete micro-components as well as cell patterning in both monolayer and three dimensional cell culture. PMID- 18432340 TI - Fast-lysis cell traps for chemical cytometry. AB - Electrically addressable cell traps were integrated with capillary electrophoresis for the analysis of the contents of single adherent cells. Electrodes composed of indium tin oxide were patterned on a glass surface followed by formation of topographical cell traps using 1002F photoresist. Single cells trapped in the holes could be lysed in less than 66 ms by applying a brief electric field (10 ms) across the electrode beneath the cell and the ground electrode placed in the aqueous media above the cell traps. The gas formed during cell lysis remained localized within the cavity formed by the 1002F photoresist. The retention of the gas in the cell trap enabled the cell traps to be coupled to an overlying capillary without blockage of the capillary. Single cells cultured in the traps were loaded with fluorescein and Oregon Green and then electrically lysed. By simultaneous application of an electric field to the capillary, the cell's contents were loaded into the capillary and electrophoretically separated. Orgeon Green and fluorescein from a single cell were fully resolved in less than two minutes. The use of a single patterned electrode beneath the 1002F cell trap yielded a simple easily fabricated design that was robust when immersed in aqueous solutions. Moreover, the design can easily be scaled up to create arrays of adherent cells for serial analyses using a single capillary or for parallel analysis by mating to an array of capillaries. Enhancing the rate of analysis of single adherent cells would enable a greater understanding of cellular physiology. PMID- 18432342 TI - A novel microfluidic driver via AC electrokinetics. AB - A novel ac electrokinetic microfluidic driver based on alternating current electro-osmosis flow induced by asymmetrically capacitance/chemistry-modulated microelectrode arrays has been successfully developed and demonstrated. Asymmetric capacitance modulation (ACM) is made of comb electrode arrays and parts of individual electrode surfaces are modulated/deposited with a SiO(2) dielectric layer. This proposed design can be utilized to shift the optimal operation frequency of maximum velocity to a higher frequency to minimize electrolytic bubble generation and enhance micropumping performance. The pumping velocity, described in this paper, is measured via the tracing of microbeads and is a function of applied potential, signal frequency, buffer concentration, and dielectric layer thickness. A maximum pumping velocity up to 290 microm s(-1) in 5 mM buffer solution with the applied potential of 10 Vpp is observed in our prototype device, and the estimated maximum flow rate is up to 26.1 microl h(-1). This is the first successful demonstration regarding bubble-free ac electrokinetic micropumping via such asymmetrically capacitance-modulated electrode arrays. Design, simulation, microfabrication, experimental result, and theoretical model are described in this paper to characterize and exhibit the performance of the proposed novel bubble-free ac electrokinetic microfluidic driver. PMID- 18432343 TI - Micropallet arrays with poly(ethylene glycol) walls. AB - Arrays of releasable micropallets with surrounding walls of poly(ethylene glycol) (PEG) were fabricated for the patterning and sorting of adherent cells. PEG walls were fabricated between the SU-8 pallets using a simple, mask-free strategy. By utilizing the difference in UV-transmittance of glass and SU-8, PEG monomer was selectively photopolymerized in the space surrounding the pallets. Since the PEG walls are composed of a cross-linked structure, the stability of the walls is independent of the pallet array geometry and the properties of the overlying solution. Even though surrounded with PEG walls, the individual pallets were detached from the array by the mechanical force generated by a focused laser pulse, with a release threshold of 6 microJ. Since the PEG hydrogels are repellent to protein adsorption and cell attachment, the walls localized cell growth to the pallet top surface. Cells grown in the microwells formed by the PEG walls were released by detaching the underlying pallet. The released cells/pallets were collected, cultured and clonally expanded. The micropallet arrays with PEG walls provide a platform for performing single cell analysis and sorting on chip. PMID- 18432344 TI - An integrated microfluidic system for long-term perfusion culture and on-line monitoring of intestinal tissue models. AB - Conventional cell-based assays in life science and medical applications can be difficult to maintain functionally over long periods. Microfluidics is an emerging technology with potential to provide integrated environments for cell maintenance, continuous perfusion, and monitoring. In this study, we developed an integrated microfluidic device with on-chip pumping and detection functionalities. The microfluidic structure in the device is divided into two independent channels separated by a semipermeable membrane on which cells are inoculated and cultured. Perfusion and fluorescence measurements of culture media for each channel can be conducted by the on-chip pumping system and optical fiber detection system. Performance of the device was examined through long-term culture and monitoring of polarized transport activity of intestinal tissue models (Caco-2 cells). The cells could be cultured for more than two weeks, and monolayer transport of rhodamine 123 was successfully monitored by on-line fluorescent measurement. This device may have applications in toxicity testing and drug screening. PMID- 18432346 TI - Numerical design of electrical-mechanical traps. AB - We present a coupled immersed interface method-boundary element method (IIM-BEM) numerical technique that predicts the behaviour of deformable cells under the effect of both hydrodynamic and electrical forces. This technique is applied to the study of a hybrid electrical-mechanical trap for single-cell trapping. We report on the effect of different combinations of electrode positions and mechanical properties of the trap on the maximum loading and unloading Reynolds numbers. We also report on the effect that cells moving with the flow have on cells which have been already trapped in a cavity. PMID- 18432345 TI - Microcirculation within grooved substrates regulates cell positioning and cell docking inside microfluidic channels. AB - Immobilization of cells inside microfluidic devices is a promising approach for enabling studies related to drug screening and cell biology. Despite extensive studies in using grooved substrates for immobilizing cells inside channels, a systematic study of the effects of various parameters that influence cell docking and retention within grooved substrates has not been performed. We demonstrate using computational simulations that the fluid dynamic environment within microgrooves significantly varies with groove width, generating microcirculation areas in smaller microgrooves. Wall shear stress simulation predicted that shear stresses were in the opposite direction in smaller grooves (25 and 50 microm wide) in comparison to those in wider grooves (75 and 100 microm wide). To validate the simulations, cells were seeded within microfluidic devices, where microgrooves of different widths were aligned perpendicularly to the direction of the flow. Experimental results showed that, as predicted, the inversion of the local direction of shear stress within the smaller grooves resulted in alignment of cells on two opposite sides of the grooves under the same flow conditions. Also, the amplitude of shear stress within microgrooved channels significantly influenced cell retainment in the channels. Therefore, our studies suggest that microscale shear stresses greatly influence cellular docking, immobilization, and retention in fluidic systems and should be considered for the design of cell based microdevices. PMID- 18432347 TI - Ultrafast active mixer using polyelectrolytic ion extractor. AB - We report on a low voltage, straight/smooth surface, and efficient active micromixer. The mixing principle is based on alternative ion depletion-enrichment using a pair of positively charged polyelectrolytic gel electrodes (pPGEs), which face each other joined by a microchannel. This system has an external AC signal source electrically connected to the pPGEs via the respective 1 M KCl solutions and Ag/AgCl electrodes. When an electric bias is applied between the two pPGEs, anions are extracted through one of the pPGEs to create a local ion-deficient region. Simultaneously, an ion-rich area appears near the other pPGE due to an inward anionic flux. As the direction of the charge flow is periodically reversed by the AC signal source, the ion depletion-enrichment regions are alternately swapped with each other on the 'push-pull' basis. The turmoil between the pPGEs quickly mixes the solutions in the microchannel without any mechanical moving part or specially machined structures. In the proposed system, both AC frequency and current density can be easily and finely controlled so that one can quickly find the optimal conditions for a given sample. The micromixer as made showed a mixing efficiency higher than 90% for sample solutions of 1 mM Rhodamine 6G and PBS at pH 7.4 when the flow rate was under 6 mm s(-1). In addition to the solution-solution mixing, the micromixer can effectively mix suspended microparticles with solution. As a representative example, rapid and efficient lysis of human red blood cells was demonstrated allowing minimal damage of the white blood cells. PMID- 18432348 TI - Millisecond treatment of cells using microfluidic devices via two-step carrier medium exchange. AB - We present herein a simple but versatile microfluidic system for the treatment of cells with millisecond chemical stimulus, by rapidly exchanging the carrier medium of cells twice in a microchannel. A technique we refer to as 'hydrodynamic filtration' has been employed for the exchange of medium, in which the virtual width of flow in the microchannel determines the size of filtered cells/particles. The treatment time of cells could be rigidly adjusted by controlling the inlet flow rates and by changing the volume of the stimulating area in the microchannel. In the experiment, two types of microdevices were designed and fabricated, and at first, the ability for carrier-medium exchange was confirmed. As an application of the presented system, we examined the influence of the treatment time of HeLa cells with Triton X-100, a non-ionic surfactant used to solubilize the cellular membrane, on cell viability, varying the average treatment time from 17 to 210 ms. Both quantitative and qualitative analyses were performed to estimate the damage on cell membrane, demonstrating that the cell viability dramatically decreased when the treatment time was longer than approximately 40 ms. The obtained results demonstrated the ability of the presented system to conduct the rapid stimulation of cells, which would be useful for the analysis of biochemical reactions at the cell surface. PMID- 18432349 TI - Fourier microfluidics. AB - We present a new experimental technique for the separation of dynamic chemical signals based on their frequency domain characteristics. Such a technique can be used to create filters that separate slow signals from fast signals from a common input flow stream. The propagation of time-varying chemical waves through networks of microfluidic channels is first examined. Mathematical models and a set of simple experiments are developed that demonstrate that short microfluidic channels behave as linear delay lines. The observed dispersive broadening and delay behavior can be explained in Fourier space in terms of corresponding phase delay, amplitude decay and characteristic transfer functions. Such delay components can be utilized to implement frequency dependent interference filters. An 8th order PDMS bandpass filter chip demonstrating these ideas was constructed. The filter chip has a central frequency of 0.17 Hz and a bandwith of 0.04 Hz at a flow rate of 4 microL h(-1). PMID- 18432350 TI - Immobilization of DNAzyme catalytic beacons on PMMA for Pb2+ detection. AB - Due to the numerous toxicological effects of lead, its presence in the environment needs to be effectively monitored. Incorporating a biosensing element within a microfluidic platform enables rapid and reliable determinations of lead at trace levels. A microchip-based lead sensor is described here that employs a lead-specific DNAzyme (also called catalytic DNA or deoxyribozyme) as a recognition element that cleaves its complementary substrate DNA strand only in the presence of cationic lead (Pb(2+)). Fluorescent tags on the DNAzyme translate the cleavage events to measurable, optical signals proportional to Pb(2+) concentration. The DNAzyme responds sensitively and selectively to Pb(2+), and immobilizing DNAzyme in the sensor permits both sensor regeneration and localization of the detection zone. Here, the DNAzyme has been immobilized on a PMMA surface using the highly specific biotin-streptavidin interaction. The strategy includes using streptavidin physisorbed on a PMMA surface to immobilize DNAzyme both on planar PMMA and on the walls of a PMMA microfluidic device. The immobilized DNAzyme retains its Pb(2+) detection activity in the microfluidic device and can be regenerated and reused. The DNAzyme shows no response to other common metal cations and the presence of these contaminants does not interfere with the lead-induced fluorescence signal. While prior work has shown lead specific catalytic DNA can be used in its solubilized form and while attached to gold substrates to quantitate Pb(2+) in solution, this is the first use of the DNAzyme immobilized within a microfluidic platform for real time Pb(2+) detection. PMID- 18432351 TI - High-sensitivity, disposable lab-on-a-chip with thin-film organic electronics for fluorescence detection. AB - We report a high-sensitivity, disposable lab-on-a-chip with a thin-film organic light-emitting diode (OLED) excitation source and an organic photodiode (OPD) detector for on-chip fluorescence analysis. A NPB/Alq3 thin-film green OLED with an active area of 0.1 cm(2) was used as the excitation source, while a CuPC/C(60) thin-film OPD with 0.6 cm(2) active area was used as a photodetector. A novel cost-effective, cross-polarization scheme was used to filter out excitation light from a fluorescent dye emission spectrum. The excitation light from the OLED was linearly polarized and used to illuminate a microfluidic device containing a 1 microL volume of dye dissolved in ethanol. The detector was shielded by a second polarizer, oriented orthogonally to the excitation light, thus reducing the photocurrent due to excitation light leakage on the detector by approximately 25 dB. The fluorescence emission light, which is randomly polarized, is only attenuated by approximately 3 dB. Fluorescence signals from Rhodamine 6G (peak emission wavelength of 570 nm) and fluorescein (peak emission wavelength of 494 nm) dyes were measured in a dilution series in the microfluidic device with emission signals detected by the OPD. A limit-of-detection of 100 nM was demonstrated for Rhodamine 6G, and 10 microM for fluorescein. This suggests that an integrated microfluidic device, with an organic photodiode and LED excitation source and integrated polarizers, can be fabricated to realize a compact and economical lab-on-a-chip for point-of-care fluorescence assays. PMID- 18432352 TI - Electrokinetic sorting and collection of fractions for preparative capillary electrophoresis on a chip. AB - A microfabricated device capable of selecting and collecting multiple components from a mixture separated by capillary electrophoresis (CE) is described. This collection is automated and can be easily controlled by a set of rules defined by an operator, enabling fast and consistent operation. The device consists of an electrokinetically steered fluidic network that can be divided into three sections: a CE part, a fractions distribution region and a set of storage channels. Sample fractions leave the CE channel and are detected in the interfacial region by fluorescence intensity measurements. If an upcoming peak is detected, separation is withheld and the potentials are reconfigured to force the fraction into one of the collection channels, where they become available for further processing or analysis. The sequence of separation and collection is repeated until all the bands of interest are captured. A mixture of three fluorescent dyes (Rhodamine 6G, Rhodamine B and Fluorescein) was used to demonstrate the principle. The components were repeatedly separated by means of CE and pooled in their respective storage channels. In comparison to previous developments, the system presented in this paper offers automatic collection of all fractions in a single run. Furthermore, it is possible to run the system in a repetitive mode for accumulative pooling if more fractionated sample is required. PMID- 18432353 TI - Gold nanoparticles for one step DNA extraction and real-time PCR of pathogens in a single chamber. AB - The optothermal properties of nanoparticles are of interest for biosensors and highly sensitive biochip applications. In this respect, the longitudinal resonance of Au nanorods was used to transform near infrared energy into thermal energy in a microfluidic chip. The resulting heat generated effectively caused pathogen lysis. Consequently the DNA was extracted out of the cell body and transferred to a PCR system. This resulted in the successful demonstration of a one step real-time PCR system for pathogen detection without removal or changing of reagents. PMID- 18432354 TI - Gas-liquid selective oxidations with oxygen under explosive conditions in a micro structured reactor. AB - The gas-liquid oxidation of cyclohexane is performed at high temperature (>200 degrees C) and pressure (up to 25 bar) using pure oxygen in a Pyrex capped silicon etched microreactor which allows convenient screen reaction conditions well above the flammability limit. PMID- 18432355 TI - Pinched flow fractionation devices for detection of single nucleotide polymorphisms. AB - We demonstrate a new and flexible microfluidic based method for genotyping single nucleotide polymorphisms (SNPs). The method relies on size separation of selectively hybridized polystyrene microspheres in a microfluidic pinched flow fractionation (PFF) device. The microfluidic PFF devices with 13 mum deep channels were fabricated by thermal nanoimprint lithography (NIL) in a thin film of cyclic-olefin copolymer (mr-I T85) on a silicon wafer substrate, and the channels were sealed by thermal polymer bonding. Streptavidin coated polystyrene microspheres with a mean diameter of 3.09 microm and 5.6 microm were functionalized with biotin-labeled oligonucleotides for the detection of a mutant (Mt) or wild-type (Wt) DNA sequence in the HBB gene, respectively. Hybridization to functionalized beads was performed with fluorescent targets comprising synthetic DNA oligonucleotides or amplified RNA, synthesized using human DNA samples from individuals with point mutations in the HBB gene. Following a stringent wash, the beads were separated in a PFF device and the fluorescent signal from the beads was analyzed. Patients being wildtypes, heterozygotes or mutated respectively for the investigated mutation could reliably be diagnosed in the PFF device. This indicates that the PFF technique can be used for accurate and fast genotyping of SNPs. PMID- 18432357 TI - Nucleic acid microarrays created in the double-spiral format on a circular microfluidic disk. AB - A microfluidic microarray that is created in the double spiral format has produced a greater density of probes than in our previous report. Using this double-spiral format together with centrifugal pumping for liquid delivery, 384 x 384 hybridization assays have been performed on one circular disk at one time, at the intersections between the spiral channels and spiral probe lines. Each sample was introduced into each inlet reservoir leading to 4 spiral channels and was analyzed independently, and so the hybridization results were self-corrected among the 4 spiral channels. In this work, fast microarray hybridizations have been successfully achieved by using both complementary oligonucleotides as well as PCR products prepared from plant fungal pathogen cultures. PMID- 18432356 TI - Coupling confocal fluorescence detection and recirculating microfluidic control for single particle analysis in discrete nanoliter volumes. AB - The recent proliferation of platforms designed to handle arrays of nano- and picolitre volumes is in response to the need to perform biological assays on discrete entities, such as single cells. However, a critical challenge associated with this trend for in vitro compartmentalization is the need for highly sensitive, yet low-volume detection platforms. In this paper, we coupled confocal fluorescence detection with recirculating microfluidic control to perform single particle DNA assays within five nL chambers. The performance of this low-volume assay was shown to match that of traditional single molecule detection platforms. However, volume requirements per measurement were nearly 3 orders of magnitude less than conventional systems, enabling future integration with lab-on-a-chip systems that require discrete or digitalized sample processing. PMID- 18432359 TI - [Local recurrence after lung cancer lung]. PMID- 18432358 TI - Enhanced microfiltration devices configured with hydrodynamic trapping and a rain drop bypass filtering architecture for microbial cells detection. AB - Ultra-fine (<1 microm) microfilters are required to effectively trap microbial cells. We designed microfilters featuring a rain drop bypass architecture, which significantly reduces the likelihood of clogging at the cost of limited cell loss. The new rain drop bypass architecture configuration has a substantially lower pressure drop and allows a better efficiency in trapping protozoan cells (Cryptosporidium parvum and Giardia lamblia) in comparison to our previous generation of a microfilter device. A modified version displaying sub-micron filter gaps was adapted to trap and detect bacterial cells (Escherichia coli), through a method of cells labeling, which aims to amplify the fluorescence signal emission and therefore the sensitivity of detection. PMID- 18432360 TI - [Local recurrence after non-small cell lung cancer surgery: prognosis variables]. AB - OBJECTIVES: Retrospectively to analyse the influence of clinical and histo pathological characteristics in the local recurrence after non-small cell lung cancer surgery. METHOD: 146 patients were included and studied during a minimum of two years. The free survival of disease function (FSD) was constructed according to the Kaplan-Meier method. The univariant comparison of the FSD by Mantel-Haenszel means of logarithmic ranks test and the multivariant analysis by Cox regression model. RESULTS: In the univariant analysis 20 months of FSD and the following predicting factors of a smaller FSD: Radiological contact of the tumour with the pleura (p=0.005), peritumoral margin affectation (p=0.001), pleural affectation (p=0.006), T stratification factor (p=0.04) and N factor (p=0.002). Other factors like age, initial symptoms, second neoplasias, radiological spiculations or cavitations, transtoracic puncion, type of surgery, peribronquial affectation, so large margin tumorlike and histological type, did not show statistically significant differences in the FSD. In the multivariant study they maintain its value prognosis the pleural affectation (p=0.0014), ganglionary affectation (p=0.02) and degree of differentiation (p=0.03). CONCLUSIONS: The most important prognosis factor of local recurrence after surgery in these patients were the infiltration of the pleura, followed of the hilio-mediastinic ganglionary affectation and the degree of histological differentiation. PMID- 18432361 TI - [Hemoglobinopathies diagnosed at the University Hospital Ntra. Sra. de Candelaria and its area of reference in Santa Cruz de Tenerife during one year]. AB - OBJECTIVE: To describe number and type of hemoglobinopathies diagnosed in our area of reference during one year as well as the origin of these patients, to have a first contact with this pathology not characterized in our province until now. MATERIAL AND METHOD: All hemoglobinopathies diagnosed in one year were reviewed, either for suspected patients, after analyzing chromatogram to quantify hemoglobin (Hb) A1c or hemogram suspicious of beta-thalassemia. The screening method was high pressure liquid chromatography (HPLC), to identify and quantify HbA2, HbS and HbC. The other variants were diagnosed at reference laboratory. RESULTS: 198 hemoglobinopathies were diagnosed, 105 after measure HbA1c. The more frequent diagnoses were beta-thalassemia trait and heterozygote HbS. 61.1% of patients are from Canary Islands, mainly from Tenerife. CONCLUSIONS: The high number of carriers found from Canary Islands, should be the start to design prospective studies on population groups, in order to know the hemoglobinopathies prevalence and to perform control and prevention official programs in the province of Santa Cruz de Tenerife. PMID- 18432362 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- 18432363 TI - [Efficiency of bortezomib and dexamethasone in relapsed multiple myeloma treatment: retrospective study in consecutive cases]. AB - INTRODUCTION: Multiple myeloma (MM) is a plasm-cell neoplasm, that is characterized by a monoclonal protein in the serum or urine. Bortezomib is an efficacy drug for the second line treatment of MM. PATIENTS AND METHOD: We conducted a retrospective study of 21 consecutive cases with refractory MM treated with bortezomib and dexamethasone as second line therapy, with the objective of analyzing the overall response rate (primary end point), the progression-free survival (PFS), the overall survival (OS), the duration of response (DR) and toxicity profile (second end points). RESULTS: In our study we found an overall response rate of 70%. With a median follow-up of 15 months, we had a median PFS of 12 months (95% CI: 2-21 months), with a median OS of 17 months (95% CI: 2-32 months), and a median DR of 9 months (95% CI: 5-13 months). Fourty-seven percent of patients had neuropathy, the 33% thrombocytopenia, 13.33% anemia and 26.66% diarrhea. CONCLUSIONS: The combination of bortezomib and dexamethasone is an effective and safe treatment in second line of refractory MM, with a manageable toxicity. PMID- 18432364 TI - [Concurrent lymphoproliferative and myeloproliferative disorders in three patients]. AB - We present two patients with a diagnosis of chronic lymphoproliferative syndrome, chronic lymphocytic leukemia B (CLL B) and lymphoplasmacytic non-Hodgkin's lymphoma (NHL), who developed chronic myeloproliferative syndrome: polycythemia vera (PV) and Philadelphia-positive chronic myeloid leukemia (CML) respectively, and a third patient with chronic myeloproliferative syndrome, polycythemia vera (PV), who developed an undefined immunophenotype cyclin D1-positive chronic lymphoproliferative syndrome. The cases included in literature are scarce, and it is not known whether some common mechanism can explain both processes' pathogeneses and the control mechanisms of one process over the other. PMID- 18432366 TI - [Myelomatous ascites]. AB - Ascites is rare in patients with multiple myeloma (MM). It may be due to diverse mechanisms, most frequently because of an increased permeability of the peritoneum or because of portal hypertension due to liver infiltration. Myelomatous ascites occurs more frequently in patients having Ig-G or Ig-A paraprotein and their prognosis is poor. It is submitted the case of a female patient aged 50 years with IgA-kappa MM, who evolved with cardiac failure (CF), plasma cells leukemia and ascites of mixed cause, because of peritoneal infiltrate of myelomatous cells, hepatic compromise and CF. A review of the different causes of ascites in patients with MM is performed. There are also summarized all myelomatous ascites cases published in the literature. Our report presents the first case of myelomatous ascites in a patient with plasma cells leukemia. PMID- 18432365 TI - [Hemoperitoenum as presentation of hepatocellular carcinoma: experience in three cases with spontaneous tumoral rupture and review of the literature]. AB - Hemoperitoneum due to spontaneous rupture of hepatocellular carcinoma (HCC) constitutes a life-threatening situation if no appropriate therapy is provided. This complication is a well-known form of HCC presentation in countries with high incidence of liver tumours, but is an unusual event in Western countries, where it has been described in 5% or less of cases with HCC. We report three patients admitted to our centre with acute hemoperitoneum secondary to non-traumatic rupture as a first manifestation of not previously diagnosed HCC. A review of the related literature is also performed. PMID- 18432367 TI - [Coxsackie virus infection associated with myositis and polyarthritis]. AB - A wide spectrum of muscle disorders caused by Coxsackie B virus, ranging from acute nonspecific myalgia to rhabdomyolisis have been described. Arthritis in not widely recognised as an either manifestation of Coxsackie virus infection, and only ten patients has been reported previously. A case of polymyositis and polyarthritis associated with primary Coxsackie B infection in a 6-year-old girl is reported. Seroconversion of IgM and IgG antibodies for Coxsackie virus B was observed coinciding with the clinical features: fever, herpangine, polyarthritis, erythematous macular rash, myalgia and muscle weakness with high levels of aldolase and creatine kinase. Electromyographic changes of myositis were found. However, histopathologically biopsied muscle demonstrated a slight inflammatory mononuclear cell infiltrate without necrosis or regeneration of muscle fibres. Polyarthritis and full muscle strength was recovered 8 and 32 weeks after the onset, respectively. PMID- 18432368 TI - [Periodic familial fever]. AB - The familial periodic fevers are Known as autoinflammatory syndromes. It is important in clinical practice to recognize these uncommon illnesses characterized by recurrent bouts of unspecific systemic symptoms associated to elevation of acute phase reactants without autoantibodies or underlying infection. The clinical suspicion supported on the molecular diagnosis represents a new perspective in relation to treatment and prognosis of these patients. PMID- 18432369 TI - [Infradiagnosis of iron deficiency in ambulatory patients due to wrong use and biochemical markers limitations: utility of reticulocyte hemoglobin content]. PMID- 18432370 TI - [Cryoglobulinemia associated to cytomegalovirus primoinfection]. PMID- 18432371 TI - [Pancreatic injuries: which approach should be taken?]. PMID- 18432372 TI - [A 28 year-old woman non-HIV with Kaposi's sarcoma, thrombocytopenic purpura and hypogammaglobulinemia]. PMID- 18432373 TI - [Perianal disease with slow course]. PMID- 18432374 TI - [Middle cerebral artery ischemic stroke as a first symptom of miliar tuberculosis]. PMID- 18432375 TI - Three potential mediators of the effects of child abuse and neglect on adulthood substance use among women. AB - OBJECTIVE: This study examined mechanisms that might account for the association between early childhood abuse and neglect, and substance use and related problems in adulthood for women. METHOD: Women with documented cases of early childhood abuse and/or neglect and matched controls were interviewed in young adulthood (mean age=29 years) and again in middle adulthood (mean age=40) (n=582). We examined the mediating effects of posttraumatic stress disorder (PTSD) symptoms, stressful life events, and delinquent and criminal behavior measured in young adulthood on substance use-related problems and illicit drug use measured in middle adulthood. RESULTS: We found that all three potential mediators mediated the effects of abuse and neglect on substance-use problems and illicit drug use. When all three mediators were considered simultaneously, only stressful life events mediated the effects of child abuse and neglect for substance use-related problems and PTSD mediated for illicit drug use. These relationships were not moderated by race/ethnicity, although the effects of abuse and neglect on the mediators differed for white and non-white women. CONCLUSIONS: These findings suggest that interventions are needed with maltreated girls to recognize and attend to their PTSD symptoms and to assist them in developing coping strategies to deal with stressful life events in an attempt to reduce risk of subsequent substance use and related problems. PMID- 18432376 TI - Sexual harassment experiences and harmful alcohol use in a military sample: differences in gender and the mediating role of depression. AB - OBJECTIVE: Researchers and clinicians alike are interested in the effects of sexual harassment on mental health, including associations with problem drinking. The aim of the current investigation was to examine depression symptoms as a mediator of the association between sexual harassment during military service and current harmful alcohol use in a sample of former military personnel, stratified by gender. METHOD: Using a cross-sectional design, 3,946 former reservists were surveyed regarding their experiences of sexual harassment in the military and their current depression symptoms and harmful alcohol use. Fifty-nine percent of the final sample were female. RESULTS: As expected, women endorsed experiencing sexual harassment more than men, and men endorsed harmful drinking more than women. Sexual harassment was associated with increased depression symptoms among both men and women; however, depression symptoms mediated the association between sexual harassment and harmful alcohol use among women only. Sexual harassment was not a significant predictor of harmful alcohol use among men. CONCLUSIONS: The associations between sexual harassment, depression symptoms, and harmful alcohol use differ between men and women in this sample. Consistent with the self medication hypothesis, sexual harassment is associated with harmful drinking among women, and this association can be accounted for by symptoms of depression. The high prevalence of harmful drinking among men and the lack of an association with sexual harassment suggest that, in this sample, men's harmful drinking is influenced by factors other than sexual harassment. PMID- 18432377 TI - Substance use in marital dyads: premarital assortment and change over time. AB - OBJECTIVE: The purpose of this study was to examine change in substance use with marriage, premarriage similarity in substance use between spouses, and the role of spouse use in predicting changes in use with marriage. METHOD: Nationally representative samples of high school seniors were followed longitudinally through age 35. The sample consisted of 2,169 adults from eight senior-year cohorts (1977-1984) from the Monitoring the Future study who completed a questionnaire at least once before their first marriage and at 2-year intervals at four consecutive points in time after marriage. RESULTS: Results indicate significant reductions in use with marriage for cigarette smoking, heavy drinking, and marijuana use. Both men and women reported reductions in all three substances following marriage. Changes in women's use followed a linear pattern. Although men's decreases in cigarette smoking and heavy drinking were initially rapid and then decelerated, their decrease in marijuana use accelerated over time. Declines in use for both genders were associated with getting married to individuals who also decreased their use. Those with higher premarriage levels of substance use experienced greater reductions in use. A significant association between respondent and spouse premarital use suggests assortative mating takes place for all three substances. CONCLUSIONS: This study affirms and further qualifies the presence of a marriage effect on substance use using multiwave and multicohort national data. Results suggest that the effects of marriage on smoking, heavy drinking, and marijuana use are related to one's own initial levels of use and the direction of change in the spouse's use. These findings have important implications for life span theoretical advances as well as interventions aimed at the marital dyad. PMID- 18432378 TI - Youthful intoxication: a cross-cultural study of drinking among German and American adolescents. AB - OBJECTIVE: This study explores whether and what differences exist between German and American adolescents on levels of alcohol use and on measures of protective factors as well as risk factors with regard to alcohol use consonant with societal and cultural differences. METHOD: A series of negative binomial regression models of adolescent alcohol use is examined with data from Germany and the United States from the 2003 European School Survey Project on Alcohol and Other Drugs and Monitoring the Future. RESULTS: Despite mean differences in drinking, findings emphasize the significance of deviant peers and perceived risks in both populations. Whereas deviant peers strongly mediate the effects of social bonds, perceived risk and opportunity influence other measures in the model only slightly. Several of the interaction terms used to assess cross cultural variation are statistically significant. CONCLUSIONS: Empirically, the current results lend support to the cross-cultural generalizability of the applied criminological theoretical constructs to German adolescents. Theoretically, the findings suggest that any explanation of cross-cultural differences regarding a specific type of problem behavior should include explicit arguments about why the supposed causes are expected to apply to that specific type of behavior in the particular cultural context. PMID- 18432379 TI - Beverage preference and risk of alcohol-use disorders: a Danish prospective cohort study. AB - OBJECTIVE: The purpose of this study was to examine whether preferred type of alcoholic beverage influences the later risk of alcohol-use disorders (AUD). METHOD: A prospective cohort study was used, comprising three updated measures of alcohol intake and covariates, and 26 years of follow-up data on 18,146 individuals from the Copenhagen City Heart Study, Denmark. The study population was linked to three different registers to detect AUD registrations. RESULTS: For both genders, wine drinking was associated with lower risk of AUD irrespective of the weekly amount of alcohol consumed. Women drinking 15-21 drinks per week of only beer and distilled spirits had a risk of 15.8 (95% confidence interval [CI]: 7.8-33.3) for AUD, whereas those whose total alcohol intake comprised more than 35% wine had a risk of 2.0 (CI: 0.7-5.2). Men drinking 15-21 drinks per week of only beer and distilled spirits had a risk of 3.1 (CI: 1.8-5.4), whereas those whose total alcohol intake comprised more than 35% wine had a risk of 0.8 (CI: 0.3-2.1). Consuming more than 35% beer increased the risk of AUD for women, whereas the percentage of distilled spirits intake did not influence the risk of AUD for either women or men. CONCLUSIONS: Individuals who include wine when they drink alcohol have lower risks of AUD, independent of the total amount of alcohol consumed. The most likely explanation of these results is that lifestyle factors and personal characteristics are associated with beverage preference. PMID- 18432380 TI - Transition from alcohol to other drugs among adolescents in Taiwan: the first drinking context matters. AB - OBJECTIVE: This study examined the extent to which youthful alcohol consumption and the initiation of tobacco, betel nut, and other illegal drugs may differ by their first drinking context in Taiwan-a society with social norms that are more tolerant of underage drinking. METHOD: In 2004, in a nationally representative sample of 11,943 school-attending youth ages 15-18, information pertaining to sociodemographic characteristics, problem behaviors, lifetime experiences of alcohol, tobacco, betel nut, and illegal drugs, as well as psychoactive, drug specific age of initiation, context at first use, average frequency, and recency of use, was assessed by anonymous questionnaires. RESULTS: Youth who had their first alcoholic beverages in entertainment settings or at friends' houses were more likely to become frequent drinkers. After adjustment for socioeconomic background and problem behaviors, having had the first drink in entertainment settings was associated with a faster progression into the initiation of illegal drugs (hazard ratio [HR]=2.5, 95% confidence interval [CI]: 1.5-4.0) and betel nut (HR=1.5, 95% CI: 1.1-2.1). CONCLUSIONS: Youthful drinking pattern and transition from alcohol to betel nut and illegal drugs may vary modestly by the first drinking context. This context may be recognized as a proxy variable to identify youth at a higher risk for alcohol and other drug problems and to devise context-based educational or prevention programs. PMID- 18432381 TI - How stable is the motive-alcohol use link? A cross-national validation of the Drinking Motives Questionnaire Revised among adolescents from Switzerland, Canada, and the United States. AB - OBJECTIVE: The aim of this study was to investigate cross-national differences (1) in the four-dimensional factor structure of drinking motives; (2) in the mean levels of enhancement, coping, social, and conformity motives; and (3) in the association of these motives with adolescent alcohol use, risky single-occasion drinking, and alcohol-related problems. METHOD: Confirmatory factor analysis, analysis of variance, and structural equation modeling were applied to sample data from Switzerland (n=5,118; mean age=15.3), Canada (n=2,557; mean age=15.7), and the United States (n=607; mean age=15.7). RESULTS: The results showed that the four-dimensional factor structure of the Drinking Motives Questionnaire Revised (DMQ-R) was structurally invariant across the three countries. Although the rank order in mean levels of motive endorsement was the same across countries (i.e., highest for social, followed by enhancement, coping, and conformity), the absolute levels of endorsement were highest in the Canadian sample, followed by the Swiss and then the U.S. sample. In all three countries, enhancement and coping motives were positively related to alcohol use and to risky drinking in particular, and coping motives were additionally related to alcohol-related problems. CONCLUSIONS: The results indicate that the DMQ-R is a valid and reliable instrument to assess drinking motives across cultures. It appears therefore that the DMQ-R is an ideal instrument for inclusion in large cross national surveys and that programs that target motives as a way to reduce risky drinking may be appropriate for different drinking cultures in different geographical locations. PMID- 18432382 TI - A social influence model of alcohol use for inner-city adolescents: family drinking, perceived drinking norms, and perceived social benefits of drinking. AB - OBJECTIVE: Social influences to drink are important predictors of adolescent drinking. This study explored a social influence model of drinking among inner city adolescents. We examined the role of family drinking and perceived drinking norms in predicting 1-year follow-up perceived social benefits of drinking and the relationship of perceived social benefits of drinking with 2-year follow-up adolescent drinking. METHOD: Participants in the present study were from the control schools of a randomized trial investigating the etiology and prevention of adolescent alcohol use. During a class period at baseline in seventh grade, participants completed a questionnaire that measured self-reported alcohol use and potential predictors. The panel sample consisted of 1,318 students from baseline (seventh grade), 1-year follow-up (eighth grade), and 2-year follow-up (ninth grade). RESULTS: Structural equation modeling found that both family drinking and perceived drinking norms affected the perceived benefits of drinking. In turn, the perceived benefits of drinking predicted subsequent drinking, controlling for earlier drinking. CONCLUSIONS: These results illuminate the importance of the perceived benefits of drinking, as well as social influences to drink, in adolescent drinking. Therefore, they should be incorporated into alcohol prevention programs. PMID- 18432383 TI - Normative misperceptions of drinking among college students: a look at the specific contexts of prepartying and drinking games. AB - OBJECTIVE: In the collegiate context, misperceptions of student drinking norms are among the most salient predictors of heavy drinking. Despite overall overestimations of peer alcohol use, misperceptions of context-specific behaviors have been infrequently studied. The present study examines students' perceptions of the high-risk behaviors of prepartying and drinking games and investigates the relationship between perceived and actual behaviors. METHOD: A sample of 524 college students completed an online assessment of actual and perceived alcohol use related to prepartying and drinking games. Quantity and frequency of overall drinking, prepartying, and drinking games were assessed for perceptions of all students at the university, as well as for male and female students separately. Questions also assessed participants' overall drinking, prepartying, and drinking game behaviors. RESULTS: Participants significantly overestimated the prepartying and drinking game behaviors of all students, male students, and female students at their university. For men, perceptions of same-sex prepartying quantity and drinking game frequency and quantity were associated with actual behavior. For women, perceptions of both same-sex and other-sex prepartying quantity were associated with actual behavior. CONCLUSIONS: These findings provide preliminary support for the association between context-specific perceived norms and actual prepartying and drinking game behaviors. Addressing these same-sex and opposite sex norms during interventions may help students reduce their own engagement in these risky behaviors. PMID- 18432385 TI - Concurrent change in alcohol and drug problems among treated adolescents over three years. AB - OBJECTIVE: Many adolescents engage in polydrug use; however, little is known about whether alcohol and other drug problems show similar posttreatment trajectories of change. We examined concurrent patterns of change for Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition, symptoms related to the use of alcohol, marijuana, and other drugs and identified predictors of the most common cross-drug patterns of change. METHOD: Adolescents (N=542) recruited from addictions treatment were assessed at baseline and at 1-and 3-year follow up. Latent class mixture modeling identified trajectories for alcohol, marijuana, and other-drug symptoms. Latent class analysis identified cross-drug patterns of change and was used to examine conduct disorder and depression as predictors of cross-drug patterns of change. RESULTS: For alcohol users, three improving groups (72%), stable-low (19%) and stable-high (6%) groups, and groups with increasing trajectories (3%) were identified. For marijuana users, an asymptomatic class (23%), two improving classes (46%), stable-low (13%) and stable-high (13%) classes, and a class with an increasing trajectory (4%) were found. For users of other drugs, groups with asymptomatic (57%), improving (20%), increasing (12%), and stable-high (11%) trajectories were identified. Latent class analysis of cross-drug patterns of change identified three subtypes representing generally concordant cross-drug patterns of change and one subtype that involved stable high marijuana problems, decreasing alcohol problems, and increasing other-drug problems. Conduct disorder was associated with greater persistence of substance problems. CONCLUSIONS: The majority of treated adolescents had similar cross-drug patterns of change for alcohol, marijuana, and other drugs; however, exceptions exist. Furthermore, adolescents with co-occurring psychopathology may benefit from continuing intervention, because they tend to report more persistent posttreatment substance-related problems. PMID- 18432384 TI - The roles of negative affect and coping motives in the relationship between alcohol use and alcohol-related problems among college students. AB - OBJECTIVE: Although studies have consistently indicated that among college students alcohol use and the likelihood of experiencing alcohol-related problems are related, it is possible that additional factors strengthen the magnitude of this relationship. The purpose of the present study was to assess the moderating effect of two such factors: negative affect and coping drinking motives. METHOD: Data were collected on 316 college students at a midsized, public university in the upper Midwest who reported using alcohol. RESULTS: Findings indicated that both negative affect and coping drinking motives moderated the alcohol use alcohol problems relationship. The three-way interaction indicated that the strongest relationship between alcohol use and alcohol-related problems existed for individuals high in both negative affect and coping drinking motives. CONCLUSIONS: This study suggests that college students high in negative affect and coping drinking motives are particularly at risk for experiencing problems as a result of their alcohol use, indicating that clinicians should consider screening for these factors when conducting alcohol-related prevention and intervention efforts. PMID- 18432386 TI - Developmental trajectories of substance use from early to late adolescence: a comparison of rural and urban youth. AB - OBJECTIVE: This study investigated differences in the development of heavy drinking and marijuana use among students in urban and rural areas and assessed whether any such differences can be accounted for by locality differences in racial/ethnic makeup, social disorganization/low social bonding, feelings of despondency and escapism, and the availability of drugs. METHOD: Drawn from 62 South Dakota middle schools involved in a drug prevention field trial, participating students were assigned to a locality category based on the location of their seventh-grade school. Schools in metropolitan areas were distinguished from schools in nonmetropolitan areas. Schools in nonmetropolitan areas were further distinguished into those in micropolitan (medium and large towns) and noncore (rural areas without towns and with small towns) areas. We used latent growth curve analysis to model the influence of locality on the development of heavy drinking and marijuana use from ages 13 to 19 and to determine whether differences in development across locality were attributable to location-based differences in race/ethnicity, social disorganization/bonding, feelings of despondency and escapism, and alcohol and marijuana availability. RESULTS: Heavy drinking increased at a faster rate among youth living in micropolitan areas compared with youth living in metropolitan areas. Marijuana use increased at a faster rate among youth living in metropolitan and micropolitan areas compared with youth living in noncore areas. Differences in the rate of change in heavy drinking were attributable to differences in the racial/ethnic composition of metropolitan and micropolitan areas. Differences in the rate of change in marijuana use were attributable to differences in residential instability and marijuana availability. CONCLUSIONS: This study underscores the diversity of drug use within rural communities, suggesting that living in a very rural area is protective against some forms of drug use but that living in a rural area that includes a medium or large town is not. PMID- 18432387 TI - The Hispanic Americans Baseline Alcohol Survey (HABLAS): rates and predictors of alcohol abuse and dependence across Hispanic national groups. AB - OBJECTIVE: The primary purpose of this article is to report 12-month prevalence rates and predictors of alcohol abuse and dependence among Mexican Americans, Cuban Americans, Puerto Ricans, and South/Central Americans living in the United States. METHOD: Using a multistage cluster sample design, a total of 5,224 individuals 18 years of age and older were selected from the household population in five metropolitan areas of the United States: Miami, New York, Philadelphia, Houston, and Los Angeles. The survey weighted response rate was 76%. Personal interviews lasting an average of 1 hour were conducted in respondents' homes either in English or Spanish. RESULTS: There is considerable heterogeneity in rates of abuse and dependence across these national groups, with Mexican American and Puerto Rican men having higher rates than Cuban American and South/Central American men. The rates of dependence for Mexican American and Puerto Rican men are also higher than those for men in the U.S. general population. Further, although the highest rates of abuse and dependence are among those in their 20s, the rate decline with age is not as strong as in the U.S. population. Thus, Hispanics at older ages (40-49, 50-59) are at considerably more risk of dependence and its health consequences than the U.S. general population. This is particularly true of Puerto Rican and Mexican American men. CONCLUSIONS: Future analysis must take this heterogeneity into consideration by conducting national group-specific analysis. Prevention efforts must also be guided by these findings, which suggest that Puerto Ricans and Mexican Americans are at higher risk for abuse, dependence, and the associated consequences than the other two groups of U.S. Hispanics. PMID- 18432388 TI - Service needs of clients in outpatient substance-use disorder treatment: a latent class analysis. AB - OBJECTIVE: The purpose of this study was to determine if there are common classes of service needs among clients in outpatient substance-use disorder treatment. METHOD: Data for this study were derived from the National Treatment Improvement Evaluation Survey. This study focused on clients in outpatient treatment (N=2,256). The majority of the subjects were male (74%). Latent class analysis was used to explore different class solutions. RESULTS: A four-class solution exhibited the best fit with the data. Based on the features of the classes, the classes were named "multiple needs," "interpersonal needs," "financial needs," and "low needs." Validation analysis showed that psychiatric problems exhibited the strongest associations with the multiple needs class. CONCLUSIONS: With service needs extending beyond substance-abuse treatment problems, a classification seems to aid in characterizing the heterogeneity of this population and is suggested of the need for packaged service approaches. PMID- 18432389 TI - Alcohol consumption and HIV-1 vaginal RNA shedding among women. AB - OBJECTIVE: The purpose of this study was to examine the association between alcohol consumption and HIV-1 vaginal shedding. METHOD: HIV-1 infected women attending a large urban HIV primary care clinic in New Orleans, LA, between June 2002 and October 2004 who underwent a gynecological exam, were 18 years of age or older, and provided informed consent were included. Subjects underwent exams and were interviewed using a computer-assisted survey at baseline, 1 month, and 3 months. RESULTS: Most of the women were black (86.4%), with a median age of 38 (range: 19-61 years). Most (56.7%) also were on antiretroviral therapy (ART; with 89.6% self-reported adherence) and had a median CD4 of 404 copies/ml and a plasma viral load of 13,400 copies/ml at baseline. The overall period prevalence of vaginal shedding (>50 copies/swab) was 24.5%, with a baseline prevalence of 27.3%. At baseline, 30% reported any alcohol consumption, 32.6% were weekly drinkers, 25.5% were heavy episodic drinkers, and 10.0% had recent moderate to heavy alcohol consumption (two or more drinks the day before the interview). Recent moderate to heavy alcohol consumption and heavy episodic drinking were both strongly and positively associated with vaginal shedding but only among women on ART. After adjusting for plasma viral load, immunosuppression, and ART adherence, recent moderate to heavy alcohol consumption was significantly associated with vaginal shedding (adjusted prevalence odds ratio=2.29, 95% confidence interval: 1.18-14.43). No association was observed for plasma viral load and alcohol consumption. CONCLUSIONS: Recent alcohol consumption was associated with expression of HIV-1 RNA in vaginal fluids of women on ART. Future research is needed to support this observation and to determine the mechanisms behind it. PMID- 18432390 TI - Embedded alcohol messages in television series: the interactive effect of warnings and audience connectedness on viewers' alcohol beliefs. AB - OBJECTIVE: This research investigates whether warning viewers about the presence of embedded messages in the content of a television episode affects viewers' drinking beliefs and whether audience connectedness moderates the warning's impact. METHOD: Two hundred fifty college students participated in a laboratory experiment approximating a real-life television viewing experience. They viewed an actual television series episode containing embedded alcohol messages, and their subsequent beliefs about alcohol consequences were measured. Experimental conditions differed based on a 2 (Connectedness Level: low vs high) x 2 (Timing of the Warning: before or after the episode) x 2 (Emphasis of Warning: advertising vs health message) design. Connectedness was measured, and the timing and emphasis of the warnings were manipulated. The design also included a control condition where there was no warning. RESULTS: The findings indicate that warning viewers about embedded messages in the content of a program can yield significant differences in viewers' beliefs about alcohol. However, the warning's impact differs depending on the viewers' level of connectedness to the program. In particular, in comparison with the no-warning control condition, the advertising prewarning produced lower positive beliefs about alcohol and its consequences but only for the low-connected viewers. Highly connected viewers were not affected by a warning emphasizing advertising messages embedded in the program, but a warning emphasizing health produced significantly higher negative beliefs about drinking than in the control condition. CONCLUSIONS: The presence of many positive portrayals of drinking and alcohol product placements in television series has led many to suggest ways to counter their influence. However, advocates of warnings should be conscious of their differential impact on high- and low connected viewers. PMID- 18432391 TI - An experimental study of the agreement of self-administration and telephone administration of the Timeline Followback interview. AB - OBJECTIVE: The Timeline Followback (TLFB) interview has become state-of-the science for the collection of retrospective self-reports of daily alcohol consumption. Such data are especially useful for addressing questions of the co occurrence of quantity of alcohol consumption and other behaviors, such as HIV related risky sex, on the event level. The purpose of this study was to determine if the TLFB could be used effectively by self-administration compared with the more costly telephone interview in a large, multisite observational study of HIV positive and HIV-negative adults. METHOD: An experimental design was used to compare self-administered and telephone-administered TLFB modes in a subsample (N=70) of the Veterans Aging Cohort Study, an ongoing longitudinal study of more than 6,000 HIV-positive and HIV-negative men and women presenting for treatment at eight Department of Veterans Affairs Infectious Disease or General Medicine clinics. Participants were randomly assigned to one of four experimental groups defined by mode and sequence of a TLFB administration on two occasions occurring within 1 week: telephone-telephone, telephone-self, self-telephone, and self self. RESULTS: Analyses showed no differences in median total number of drinks reported between modes of TLFB administration or sequence of mode of administration. The same findings held for classification of participants as "hazardous" drinkers. Additional analyses showed good-to-excellent test-retest reliability of self-reports for both modes of TLFB administration. CONCLUSIONS: The data derived from this study provide strong experimental evidence for the utility of the self-administered, 30-day TLFB in collecting daily alcohol consumption in large observational studies of HIV-positive and HIV-negative individuals. PMID- 18432392 TI - Argileh use among college students in the United States: an emerging trend. AB - OBJECTIVE: This study examined the prevalence and predictors of argileh (hookah pipe) use among a sample of nonselected college students. METHOD: Participants were 602 students (24% male; 43% white; mean age=22.06) at a large, ethnically diverse, urban university. All participants completed an online survey designed to assess various types of substance use. RESULTS: More than 15% of the sample reported having used argileh at least once in their lifetime, exceeding the percentage of students who had tried stimulants, barbiturates, cocaine, Ecstasy, heroin, or psychedelics. Arab ethnicity and cigarette smoking were the strongest predictors of argileh use; however, a substantial percentage of non-Arabs and nonsmokers also had tried argileh. CONCLUSIONS: Findings suggest that, in comparison with other substances, the prevalence of argileh use is high among college students in the United States. Physical health implications of these findings are discussed. PMID- 18432393 TI - Centromere repositioning in the X chromosome of XO/XO mammals, Ryukyu spiny rat. AB - Two species of Ryukyu spiny rat, Tokudaia osimensis and Tokudaia tokunoshimensis, have an XO/XO sex chromosome constitution with no cytogenetically visible Y chromosome in both sexes. The single X chromosomes of T. osimensis and T. tokunoshimensis are submetacentric and subtelocentric, respectively. It was therefore suggested that a pericentric inversion event occurred in the X chromosome of either species. To identify X chromosome rearrangements that have occurred between the two species, we mapped 22 mouse cDNA clones of the X-linked genes on the chromosomes of the two species by direct R-banding FISH. The gene orders of the X chromosomes were conserved in the two species, whereas the position of the centromere on the X chromosome was different. This result indicates that the rearrangement which occurred in either of the X chromosomes after the two species diverged from a common ancestor involved not pericentric inversion but centromere repositioning. PMID- 18432394 TI - Cardiac positron emission tomography: myocardial perfusion and metabolism in clinical practice. PMID- 18432395 TI - Combined endurance and muscle strength training in female and male patients with chronic heart failure. AB - BACKGROUND: Exercise training has now become established practice in patients with chronic heart failure. Women are often under-represented in intervention studies compared to men. For this reason it was our aim to conduct a combined endurance and muscle strength training program to evaluate its effect on clinical performance data and health-related psychosocial factors in women and men. METHODS: One hundred and sixteen women, mean age 69 +/- 9 years, body mass index (BMI) 25.8 +/- 4.9, and 169 men, mean age 66 +/- 9 years, BMI 26.6 +/- 3.6 underwent combined endurance/resistance training. The training program lasted 29 +/- 7 days and comprised bicycle ergometer training, a 6-min walk test as a training unit and muscle strength training for the lower and upper extremities. RESULTS: Differences between women and men were found in clinical parameters. In particular, statistically significant differences were revealed between the women and men with regard to cardiopulmonary performance. Quality of life was significantly improved on discharge with regard to both physical and mental health, whereas anxiety and depression showed no significant alteration. CONCLUSION: A specialized in-hospital program for women and men combining endurance/resistance training and education is feasible. But our program revealed a very low level of cardiopulmonary performance in women. Women need to be encouraged and motivated to participate in such programs. PMID- 18432396 TI - Depressive symptoms in elementary school children in Jeju Island, Korea: prevalence and correlates. AB - We investigated the prevalence and correlates of depressive symptoms in elementary school children in Jeju Island, Korea. The study participants were 2305 children enrolled in elementary schools in Jeju-si, Seogwipo-si, Namjeju gun, and Bukjeju-gun and their parents who completed questionnaires about sociodemographics, health habits, family relationship information, and the Korean form of the Kovac's children's depression inventory (CDI) in September to December 2005. Multiple logistic regression showed that higher age (OR = 1.259, 95% CI 1.098-1.445), short time spent developing a relationship with the mother (OR = 2.770, 95% CI 1.280-5.944), and a low level of body image satisfaction (OR = 3.397, 95% CI 1.823-6.330) were correlates of depressive symptoms in children. Our results suggest that the following are essential to prevent depressive symptoms in elementary school children in Jeju, Korea: advanced education and social activity programs at home, in school, and in the community to help children have a positive self-image, and much time spent building a relationship with the mother. PMID- 18432397 TI - A comparative study on the efficiency of two enucleation methods in pig somatic cell nuclear transfer: effects of the squeezing and the aspiration methods. AB - In this study, two enucleation methods, the squeezing and the aspiration methods, were compared. The efficiency of these two methods to enucleate pig oocytes and the in vitro and in vivo viability of somatic cell nuclear transfer (SCNT) pig embryos, were evaluated. In the squeezing method, the zona pellucida was partially dissected and a small amount of cytoplasm containing metaphase II (MII) chromosomes and the first polar body (PB) were pushed out. In the aspiration method, the PB and MII chromosomes were aspirated using a beveled micropipette. After injection of fetal fibroblasts into the perivitelline space, reconstructed oocytes were fused and activated electrically, and then cultured in vitro for 6 days or transferred to surrogates. The squeezing method resulted in a higher proportion of degenerated oocytes than the aspiration method (14% vs. 5%). The squeezing method took longer to enucleate 100 oocytes (306 minutes) than the aspirating method (113 minutes). Fusion rate (72-78%) and cleavage rate (67%) were not influenced by the enucleation method but blastocyst formation was improved (P < 0.05) in oocytes enucleated by the aspiration method (5 vs. 9%). When SCNT embryos were transferred to recipients, pregnancy rates to term were similar (27%, 3/11 and 27%, 3/11) in both methods with the birth of 10 piglets/3 litters and 16 piglets/3 litters in the squeezing and the aspiration methods, respectively. Our results indicate that the aspiration method for oocyte enucleation is more efficient than the squeezing method in producing a large number of pig SCNT embryos with normal in vivo viability. PMID- 18432398 TI - Association and in silico assignment of sequences from turkey BACs. AB - Bacterial artificial chromosomes (BACs) provide an important resource in genetic mapping. An initial set of BACs corresponding to microsatellite markers in the turkey (Meleagris gallopavo) was isolated from the CHORI-260 turkey BAC library. The selected markers were distributed on both macro- and microchromosomes and included a genetically unlinked marker. End sequences were obtained for a subset of the recovered BACs and compared to the chicken whole genome sequence. Close association of the turkey BAC-end sequences and original marker sequences was generally conserved in the chicken genome. Gene content of the turkey BACs is predicted from the comparative sequence alignments. PMID- 18432399 TI - Development of cell lines from the sheep used to construct the CHORI-243 Ovine BAC library. AB - Two cell lines, designated MARC.OVSM, and MARC.OKF, were initiated from the aorta and kidney, respectively, obtained from the Texel ram used to make the CHORI-243 Ovine BAC library. These cell lines have been submitted to the NIA Aging Cell Repository at the Coriell Cell Respositories, Camden, NJ, USA, and will be made publicly available. PMID- 18432400 TI - WDNM1 is associated with differentiation and apoptosis of mammary epithelial cells. AB - In this study, we show that expression of the Westmead DMBA8 nonmetastatic cDNA 1 (WDNM1) gene was increased upon SFM and/or TNFalpha treatment, with a corresponding increase in apoptotic cells, and gradually decreased following re stimulation with serum in HC11 mammary epithelial cells. TNFalpha induced WDNM1 expression showed the NFkappaB-dependent mechanism since it's expression was abrogated in IkappaBalphaM (super-repressor of NFkappaB)-transfected cells, but not those transfected with control vector. Furthermore, overexpression of WDNM1 suppressed growth and differentiation, and accelerated apoptosis of HC11 cells. Thus, our results demonstrate that WDNM1 gene expression, regulated by the TNFalpha-NFkappaB signal pathway, is associated with HC11 cell apoptosis. PMID- 18432401 TI - Simultaneous expression of growth hormone releasing hormone (GHRH) and hepatitis B surface antigen/somatostatin (HBsAg/SS) fusion genes in a construct in the skeletal muscle enhances rabbit weight gain. AB - Somatostatin (SS) and growth hormone-releasing hormone (GHRH) are synthesized and secreted by the hypothalamus, which can control the synthesis and secretion of the growth hormone (GH) from the hypophysis as well as regulate the GH concentrations in animals and humans. In this article, we describe the regulation of animal growth using plasmid DNA encoding both the GHRH gene and the SS gene fused with the hepatitis B surface antigen (HBsAg) gene. We constructed a series of expression plasmids to express the GHRH and HBsAg-SS fusion genes individually as well as collectively. The fusion gene and GHRH were successfully expressed in Chinese hamster ovary (CHO) cells, as proven by reverse transcriptase-polymerase chain reaction (RT-PCR) and immunoblotting tests. Poly D, L-lactide-co-glycolic acid (PLGA) plasmid-encapsulating microspheres were prepared and injected intramuscularly into the leg skeletal muscles of rabbits. Weight gain/day and the levels of insulinlike growth factor-I (IGF-I), SS, and hepatitis B surface antibody (HBsAb) were monitored. During days 30 postinjection, increase in weight gain/day and IGF- I concentration and decrease in SS were observed in treatment groups. From days 15 to 30 postinjection, the weight gain/day significantly increased (P < 0.05) by 129.13%, 106.8%, and 72.82% relative to the control group in the co-expression GHRH and fusion gene (named P-G-HS), fusion gene (named P HS), and GHRH (named P-G) groups, respectively. And most importantly, the P-G-HS group showed significant weight gain/day (P < 0.05) relative to the P-G and P-HS groups. A significant increase in the IGF-I concentration and decrease in the SS level relative to the control group were also observed. The results indicated that the combination of plasmid-mediated GHRH supplementation and positive immunization against SS led to more robust weight gain/day in rabbits. PMID- 18432402 TI - cDNA cloning and tissue expression analyses of the encoding regions for three novel porcine genes- MJD1, CDC42 and NECD. AB - The cDNAs for Machado-Joseph disease protein 1 homolog (MJD1), cell division control protein 42 homolog precursor(CDC42) and necdin (NECD) genes of pig were amplified using the reverse transcriptase polymerase chain reaction (RT-PCR) based on the conserved coding sequence information of the MJD1, CDC42, and NECD genes from mouse and other mammals and the referenced porcine EST sequence information. Tissue expression analysis showed the swine MJD1, CDC42, and NECD genes were obviously differentially expressed in different tissues including muscle, heart, liver, backfat, kidney, lung, small intestine, and large intestine. Our experiment established the primary foundation for further research on these three swine genes. PMID- 18432404 TI - Epigenetics and autoimmunity. Overview. PMID- 18432403 TI - A new tandem repeat in bovine fibrinogen Aalpha gene. AB - In this study, we describe intraspecies variation in the alphaC connector region of the bovine fibrinogen Aalpha gene. Sequencing and genotyping of six bovine breeds revealed 7 to 10 tandem repeats in the alphaC connector region. In addition, we observed length differences between B. indicus and B. taurus, with the B. indicus having longer fibrinogen alphaC connectors (10-repeat alleles) than B. taurus (7- and 9-repeats). The difference in tandem repeats may be related to the function of blood coagulation system. PMID- 18432405 TI - Epigenetics and T-cell immunity. AB - Epigenetic mechanisms including DNA methylation and histone modifications are critically involved in immune responses. Antigen stimulation along with a specific cytokine milieu drives helper T-cell differentiation into specific subsets with distinct functional capacities. This process occurs by inducing chromatin remodeling and altering transcriptional accessibility of key cytokine genes such as IFN-gamma, IL-4 and IL-17. These epigenetic changes, by definition, are carried over throughout cell division to ensure selective survival of a cell lineage. Over the past decade, a growing body of literature mechanistically uncovered the central role for epigenetic regulation in immunity. In this review, we focus on epigenetics in T helper cell differentiation, regulatory T-cell function, and IL-2 production. PMID- 18432407 TI - Skewing of X chromosome inactivation in autoimmunity. AB - Approximately 5% of the population in Western countries is affected by autoimmune diseases (AID), with a significantly higher prevalence in women. Genetic factors are known to be crucial determinants of susceptibility as shown by family and twin studies, although no specific genes predisposing women to autoimmunity have been identified thus far. Several studies indicate that X chromosome abnormalities, such as inactivation patterns, characterize some female predominant AID. We herein review the most recent evidence on the role of the X chromosome in the breakdown of immune tolerance and discuss its potential implications. Future efforts will help to identify specific X chromosome regions containing candidate genes for disease susceptibility. PMID- 18432406 TI - ICF, an immunodeficiency syndrome: DNA methyltransferase 3B involvement, chromosome anomalies, and gene dysregulation. AB - The immunodeficiency, centromeric region instability, and facial anomalies syndrome (ICF) is the only disease known to result from a mutated DNA methyltransferase gene, namely, DNMT3B. Characteristic of this recessive disease are decreases in serum immunoglobulins despite the presence of B cells and, in the juxtacentromeric heterochromatin of chromosomes 1 and 16, chromatin decondensation, distinctive rearrangements, and satellite DNA hypomethylation. Although DNMT3B is involved in specific associations with histone deacetylases, HP1, other DNMTs, chromatin remodelling proteins, condensin, and other nuclear proteins, it is probably the partial loss of catalytic activity that is responsible for the disease. In microarray experiments and real-time RT-PCR assays, we observed significant differences in RNA levels from ICF vs. control lymphoblasts for pro- and anti-apoptotic genes (BCL2L10, CASP1, and PTPN13); nitrous oxide, carbon monoxide, NF-kappaB, and TNFalpha signalling pathway genes (PRKCH, GUCY1A3, GUCY1B3, MAPK13; HMOX1, and MAP4K4); and transcription control genes (NR2F2 and SMARCA2). This gene dysregulation could contribute to the immunodeficiency and other symptoms of ICF and might result from the limited losses of DNA methylation although ICF-related promoter hypomethylation was not observed for six of the above examined genes. We propose that hypomethylation of satellite 2 at 1qh and 16qh might provoke this dysregulation gene expression by trans effects from altered sequestration of transcription factors, changes in nuclear architecture, or expression of noncoding RNAs. PMID- 18432408 TI - Epigenetics in human autoimmunity. Epigenetics in autoimmunity - DNA methylation in systemic lupus erythematosus and beyond. AB - Epigenetic mechanisms are essential for normal development and function of the immune system. Similarly, a failure to maintain epigenetic homeostasis in the immune response due to factors including environmental influences, leads to aberrant gene expression, contributing to immune dysfunction and in some cases the development of autoimmunity in genetically predisposed individuals. This is exemplified by systemic lupus erythematosus, where environmentally induced epigenetic changes contribute to disease pathogenesis in those genetically predisposed. Similar interactions between genetically determined susceptibility and environmental factors are implicated in other systemic autoimmune diseases such as rheumatoid arthritis and scleroderma, as well as in organ specific autoimmunity. The skin is exposed to a wide variety of environmental agents, including UV radiation, and is prone to the development of autoimmune conditions such as atopic dermatitis, psoriasis and some forms of vitiligo, depending on environmental and genetic influences. Herein we review how disruption of epigenetic mechanisms can alter immune function using lupus as an example, and summarize how similar mechanisms may contribute to other human autoimmune rheumatic and skin diseases. PMID- 18432410 TI - Regulation and dysregulation of Epstein-Barr virus latency: implications for the development of autoimmune diseases. AB - Epstein-Barr virus (EBV) is a human herpesvirus hiding in a latent form in memory B cells in the majority of the world population. Although, primary EBV infection is asymptomatic or causes a self-limiting disease, infectious mononucleosis, the virus is associated with a wide variety of neoplasms developing in immunosuppressed or immunodeficient individuals, but also in patients with an apparently intact immune system. In memory B cells, tumor cells, and lymphoblastoid cell lines (LCLs, transformed by EBV in vitro) the expression of the viral genes is highly restricted. There is no virus production (lytic viral replication associated with the expression of all viral genes) in tight latency. The expression of latent viral oncogenes and RNAs is under a strict epigenetic control via DNA methylation and histone modifications that results either in a complete silencing of the EBV genome in memory B cells, or in a cell-type dependent usage of latent promoters in tumor cells, germinal center B cells, and LCLs. Both the latent and lytic EBV proteins are potent immunogens and elicit vigorous B- and T-cell responses. In immunosuppressed and immunodeficient patients, or in individuals with a functional defect of EBV-specific T cells, lytic EBV replication is regularly activated and an increased viral load can be detected in the blood. Enhanced lytic replication results in new infection events and EBV-associated transformation events, and seems to be a risk factor both for malignant transformation and the development of autoimmune diseases. One may speculate that an increased load or altered presentation of a limited set of lytic or latent EBV proteins that cross-react with cellular antigens triggers and perpetuates the pathogenic processes that result in multiple sclerosis, systemic lupus erythematosus (SLE), and rheumatoid arthritis. In addition, in SLE patients EBV may cause defects of B-cell tolerance checkpoints because latent membrane protein 1, an EBV-encoded viral oncoprotein can induce BAFF, a B-cell activating factor that rescues self-reactive B cells and induces a lupus-like autoimmune disease in transgenic mice. PMID- 18432409 TI - Molecular mimicry and immunomodulation by the HRES-1 endogenous retrovirus in SLE. AB - Genetic and environmental factors are believed to influence development of systemic lupus erythematosus (SLE). Endogenous retroviruses (ERV) correspond to the integrated proviral form of infectious retroviruses, which are trapped within the genome due to mutations. ERV represent a key molecular link between the host genome and infectious viral particles. ERV-encoded proteins are recognized by antiviral immune responses and become targets of autoreactivity. Alternatively, ERV protein may influence cellular processes and the life cycle of infectious viruses. As examples, the HRES-1 human ERV encodes a 28-kDa nuclear autoantigen and a 24-kDa small GTP-ase, termed HRES-1/Rab4. HRES-1/p28 is a nuclear autoantigen recognized by cross-reactive antiviral antibodies, while HRES-1/Rab4 regulates surface expression of CD4 and the transferrin receptor (TFR) through endosome recycling. Expression of HRES-1/Rab4 is induced by the tat gene of HIV 1, which in turn down-regulates expression of CD4 and susceptibility to re infection by HIV-1. CD4 and the TFR play essential roles in formation of the immunological synapse (IS) during normal T-cell activation by a cognate MHC class II peptide complex. The key intracellular transducer of T-cell activation, Lck, is brought to the IS via binding to CD4. T-cell receptorzeta (TCRzeta) chain binds to the TFR. Abnormal T-cell responses in SLE have been associated with reduced lck and TCRzeta chain levels. HRES-1 is centrally located on chromosome 1 at q42 relative to lupus-linked microsatellite markers and polymorphic HRES-1 alleles have been linked to the development of SLE. 1q42 is one of the three most common fragile sites in the human genome, and is inducible by DNA demethylation, a known mechanism of retroviral gene activation. Molecular mimicry and immunomodulation by a ERV, such as HRES-1, may contribute to self-reactivity and abnormal T and B-cell functions in SLE. PMID- 18432412 TI - Respiratory motion compensation for CT-guided interventions in the liver. AB - Computed tomography (CT) guided minimally invasive procedures in the liver, such as tumor biopsy and thermal ablation therapy, require precise targeting of hepatic structures that are subject to breathing motion. To facilitate needle placement, we introduced a navigation system which uses needle-shaped optically tracked navigation aids and a real-time deformation model to continuously estimate the position of a moving target. In this study, we assessed the target position estimation accuracy of our system in vitro with a custom-designed respiratory liver motion simulator. Several real-time compatible transformations were compared as a basis for the deformation model and were evaluated in a set of experiments using different arrangements of three navigation aids in two porcine and two human livers. Furthermore, we investigated different placement strategies for the case where only two needles are used for motion compensation. Depending on the transformation and the placement of the navigation aids, our system yielded a root mean square (RMS) target position estimation error in the range of 0.7 mm to 2.9 mm throughout the breathing cycle generated by the motion simulator. Affine transformations and spline transformations performed comparably well (overall RMS < 2 mm) and were considerably better than rigid transformations. When two navigation aids were used for motion compensation instead of three, a diagonal arrangement of the needles yielded the best results. This study suggests that our navigation system could significantly improve the clinical treatment standard for CT-guided interventions in the liver. PMID- 18432413 TI - An integer programming model for distal humerus fracture fixation planning. AB - OBJECTIVE: To demonstrate the feasibility of an integer programming model to assist in pre-operative planning for open reduction and internal fixation of a distal humerus fracture. MATERIALS AND METHODS: We describe an integer programming model based on the objective of maximizing the reward for screws placed while satisfying the requirements for sound internal fixation. The model maximizes the number of bicortical screws placed while avoiding screw collision and favoring screws of greater length that cross multiple fracture planes. RESULTS: The model was tested on three types of total articular fractures of the distal humerus. Solutions were generated using 5, 9, 21 and 33 possible screw orientations per hole. Solutions generated using 33 possible screw orientations per hole and five screw lengths resulted in the most clinically relevant fixation plan and required the calculation of 1,191,975 pairs of screws that resulted in collision. At this level of complexity, the pre-processor took 104 seconds to generate the constraints for the solver, and a solution was generated in under one minute in all three cases. CONCLUSION: Despite the large size of this problem, it can be solved in a reasonable amount of time, making use of the model practical in pre-surgical planning. PMID- 18432411 TI - Epigenetics, aging, and autoimmunity. AB - Immune senescence is associated with a decline in T- and B-cell immune responses. It is, therefore, perhaps surprising that aging is linked to the appearance of serological and clinical autoimmunity. Here we review the mechanisms that contribute to the increase in inflammatory and autoimmune responses in aging. The bulk of this review will focus on aging-associated changes in epigenetic mechanisms, and in particular DNA methylation, as this has emerged as an attractive mechanistic link between aging and autoimmunity. PMID- 18432414 TI - Robot-assisted femoral fracture reduction: preliminary study in patients and healthy volunteers. AB - We developed a robot-assisted fracture reduction system (FRAC-Robo) to assist anatomical reduction and to maintain reduction during internal fixation while recording the procedure in a log. We conducted two experiments before using FRAC Robo clinically. In the first experiment using the FRAC-Robo system, we measured the maximum force and torque required to pull and rotate the limbs of healthy conscious volunteers until they felt pain or abnormality. The average maximum traction force applied to the lower limb was 250.7 N, and the average maximum torque was 5.6 Nm in internal rotation and 7.6 Nm in external rotation for 30 degrees of abduction of hip. In the second experiment, we measured the traction force and rotation torque during the reduction of proximal femoral fractures. The average traction force and rotation torque needed for reduction were 215.9 N and 3.2 Nm, respectively. On the basis of these results, we consider that FRAC-Robo can generate sufficient force and torque to reduce femoral fractures safely. PMID- 18432415 TI - Navigated pedicle screw placement: experimental comparison between CT- and 3D fluoroscopy-based techniques. AB - OBJECTIVE: Even with CT-based navigation, the misplacement rate for pedicle screws is reported to be as high as 10%. Using fluoroscopy-based 3D navigation, misplacement rates of 1.7 to 6% occur. The purpose of this study was to compare the accuracy of CT-based and Iso-C-based navigation in an experimental context. METHODS: A foam spine model and the SurgiGATE navigation system were used. First, a determination of point accuracy measured the difference between the real positions of markers placed on selected vertebrae and their positions as determined by the navigation system. In the verification mode, the pointer is placed exactly on the markers displayed on the monitor screen, and the deviation of the pointer tip and marker is measured in reality using a caliper. Secondly, pedicle accuracy was measured using pre-drilled holes for pedicle screws. A trajectory was planned into the visible hole and the navigated pointer was placed. RESULTS: The measured accuracy for the markers showed a statistically significant difference between the results with CT and Iso-C navigation for one of six markers placed on the vertebra. Iso-C-based navigation demonstrated a lower mean deviation of 0.5 mm, compared to 1 mm with CT-based navigation. The deviation within the pre-drilled holes was lower when using the Iso-C3D scan. Using Iso-C3D navigation, 76.6% of the measurements showed no deviation at the entrance point, compared with 43% when using CT-based navigation. Also, with Iso C3D navigation, 78.3% of the inserted pedicle awls hit the defined trajectories in the pre-drilled holes correctly, compared to 66.6% with CT-based navigation. CONCLUSION: The overall image-to-reality accuracy for CT- and Iso-C-based navigation was assessed in the described experimental setup. An apparent tendency towards higher accuracy with Iso-C-based navigation was evaluated; however, the differences were not significant. PMID- 18432416 TI - Accuracy of femoral rotational alignment in total knee arthroplasty using computer assisted navigation. AB - Proper rotational alignment of components is crucial for the success of total knee arthroplasty (TKA). The traditional reference guides for assessment of femoral rotation include the posterior condylar axis, the anteroposterior axis, also known as Whiteside's line, and the transepicondylar axis (TEA). The fixed angle posterior referencing system recommends that the rotational femoral cut be made at 3 degrees of external rotation. In a consecutive series of 100 patients undergoing TKA at our institution, the accuracy of these reference guides in determining the rotation of the femoral component was compared with that of a computerized navigation system. Although differences between the three reference methods were not statistically significant, the possibility of finding an outlier leading to excessive external or internal rotation of the femoral component when using a fixed posterior condyle reference guide mandates the use of other referencing methods to avoid this error. Using fixed posterior referencing, up to 17% of femoral components would have differed by more than 5 degrees from the anatomic reference landmarks (TEA, Whiteside's line). This degree of rotational malalignment could lead to knee instability and early failure. PMID- 18432417 TI - Navigated non-image-based registration of the position of the pelvis during THR. An accuracy and reproducibility study. AB - The precise recording of the position of the pelvis is a prerequisite for total hip replacement (THR). The anterior pelvic plane is an accepted reference for determining the 3D pelvic orientation. We hypothesized that cutaneous palpation of this plane was accurate and reproducible. Ten consecutive navigated implantations of THR prostheses were studied. In each case, four palpations of both anterior iliac spines and the pubic symphysis were performed with a navigated stylus. The first palpation was made on actual bone contours through a skin puncture and was considered as the reference. The other three palpations were made over the intact skin as a normal intra-operative procedure. There was no significant difference between the pelvic orientations measured by the three cutaneous palpations, or between the orientations measured by transcutaneous palpation and the mean result with cutaneous palpation. Cutaneous palpation can be considered as a reliable technique for the definition of pelvic orientation during THR with the non-image-based system employed. PMID- 18432418 TI - Neuroscience and hormesis: overview and general findings. AB - This article provides a summary of an assessment of the occurrence and impact of hormesis in the neurosciences, including the areas of neuroprotection, neurite outgrowth, and drugs for Alzheimer's disease, Parkinson's disease, anxiety, pain, seizures, stroke, as well as in the areas of behavioral pharmacology, addictive drugs, stress biology including the Yerkes-Dodson law, and p-glycoprotein efflux activity. The findings indicate that the hormetic dose response has a common, if not dominant, presence in each of these diverse areas of neuroscience and further strengthens the conclusion that hormesis is highly generalizable, being independent of biological model, endpoint, and chemical class. PMID- 18432419 TI - Dose-response features of neuroprotective agents: an integrative summary. AB - This article provides an integrative summary of the effects of neuroprotective agents on neuronal survival and neurite outgrowth using primary cell cultures, multiple neuronal cell lines, and astroglial cells. These findings are dealt with in considerable detail in the following three articles (Calabrese, 2008a, 2008b, 2008c) of this series of issues of Critical Reviews in Toxicology. The principal finding is that the overwhelming majority of neuroprotective agents display biphasic dose responses, characterized by modest low-dose enhancement/stimulation and high-dose inhibitory responses. The quantitative features of these dose responses are consistent with the hormetic dose-response model. Mechanisms that account for numerous hormetic dose responses of neuroprotective agents are summarized, as well as the clinical implications of specific experimental findings. PMID- 18432420 TI - Pharmacological enhancement of neuronal survival. AB - This article is a comprehensive assessment of the quantitative features of the dose response for neuroprotective agents and their underlying mechanistic foundations. The data were derived from published studies using numerous primary neuronal cell cultures and neuronal cell lines. These biological models assessed normal developmental and aging processes, preconditioning adaptive responses, and various neurodegenerative diseases, including Alzheimer's disease, Parkinson's disease, and Huntington's disease. The nature of the dose response was generally U-shaped with quantitative features similar to the hormetic dose-response model and independent of biological model, endpoint measured, and chemical class. The agents displaying the U-shaped dose response for neuronal survival include numerous endogenous agonists, plant-derived agents, synthetic drugs, and widely used chemicals including potent neurotoxins. That neuroprotective agents display similar dose-response relationships regardless of experimental system, potency, and endpoint is an important observation with widespread biomedical and clinical implications. PMID- 18432421 TI - Enhancing and regulating neurite outgrowth. AB - Numerous agents have demonstrated the potential to enhance neuronal repair following spinal cord or peripheral nerve injury using neurite outgrowth as a biomarker for axonal extension in primary cell cultures and neuronal cell lines. This article provides an assessment of the dose-response features of chemically induced neuronal outgrowth in a broad range of experimental models during normal developmental processes, following chemically induced neuronal damage or processes that simulate such damage. These findings indicate that endogenous and exogenous agents, independent of biological model, stimulate central and peripheral nervous system neuronal outgrowths in a biphasic manner consistent with the quantitative features of the hormetic dose-response model. These findings have important clinical implications as they define the plasticity of neurite outgrowth stimulatory responses with respect to the magnitude of enhancement and width of the possible therapeutic zone. The findings also display an essential role for hormetic dose-response relationships in normal neuronal based developmental and tissue repair processes. PMID- 18432422 TI - Children in planned lesbian families: stigmatisation, psychological adjustment and protective factors. AB - The study assessed the extent to which children between eight and 12 years old in planned lesbian families in the Netherlands experience stigmatization, as well as the influence of protective factors (relationship with parents, social acceptance by peers, contact with children from other families headed by lesbian mothers or gay fathers) on their psychological adjustment (conduct problems, emotional symptoms, hyperactivity, self-esteem). Data were collected by questionnaires filled out by the mothers and by the children themselves. The children in the sample generally reported low levels of stigmatization. However, boys more often reported that, in their view, they were excluded by peers because of their non traditional family situation. Girls more often reported that other children gossiped about the fact that they had two lesbian mothers. Higher levels of stigmatization were associated with more hyperactivity for boys and lower self esteem for girls. Having frequent contact with other children who have a lesbian mother or gay father protects against the negative influence of stigmatization on self-esteem. Findings support the idea that children in planned lesbian families benefit from the experience of meeting other children from similar families. PMID- 18432423 TI - Sharing beliefs: what sexuality means to Muslim Iranian women living in Australia. AB - In Iran, women's sexual self-understandings are strongly determined by religious teaching. This study explores the meanings generated through the lived experience of sexuality of women residing in Australia that may challenge certain received Islamic notions of sexuality. Interviews and focus group discussions were conducted with 51 Iranian women in Sydney alongside 10 semi-structured interviews with Shi'ite clergy in Iran. Findings suggest that the concept of sexual obedience within marriage is regarded as symbolic of an idealised Muslim femininity. Sexual obedience demonstrates women's high level of religious commitment but is also an indicator of modesty and self-respect. While some participants felt that it was a woman's duty to satisfy her husband on any sexual occasion, they did not utilize notions of subordination in their sexual lives. For other informants, however, concepts of subordination were more salient and tied to conservative religious leaders' interpretations of Quranic concepts and Iranian women's understandings of these same interpretations. Religion is an important factor influencing Iranian women's sexual self-understandings, being relevant both to sexual education and public health. Recognition of this issue will facilitate understanding of the cultural foundations of sexuality among Muslims and assist health providers in suggesting more culturally compatible forms of healthcare. PMID- 18432424 TI - Building stakeholder partnerships for an on-site HIV testing programme. AB - Because of the large number of individuals at risk for HIV infection who visit gay saunas and sex clubs, these venues are useful settings in which to offer HIV outreach programmes for voluntary counselling and testing (VCT). Nevertheless, establishing a successful VCT programme in such a setting can be a daunting challenge, in large part because there are many barriers to managing the various components likely to be involved. Using qualitative data from a process evaluation of a new VCT programme at a gay sauna in California, USA, we describe how the various stakeholders overcame barriers of disparate interests and responsibilities to work together to successfully facilitate a regular and frequent on-site VCT programme that was fully utilized by patrons. PMID- 18432425 TI - M4M chat rooms: individual socialization and sexual autonomy. AB - This paper uses data from twenty-one online and in-person qualitative interviews to examine the meaning and use of chat rooms located on men for men (M4M) websites from the perspectives of men seeking men on the Internet. This research is inspired by recent public health and social sciences literature on gay websites and chat rooms. The data indicate that these online sites help expedite learning about sex and sexuality and, for men who are shy or geographically isolated, to interact with metropolitan gay communities. There is, however, a measure of stigma associated with use of these chat rooms, particularly by men who are older or in coupled relationships. Using these data, the paper argues that M4M chat rooms play a vital role in fostering the sexual autonomy of many men who frequent these venues and that sociologists should devote more study to the complexity of online social interaction. PMID- 18432426 TI - Social and cultural dimensions of pregnancy and childbirth in eastern Turkey. AB - This study offers an in-depth analysis of women's perceptions and beliefs about pregnancy and delivery in a rural area of eastern Turkey. The goal was to identify possible threats to reproductive health among women in rural, conservative communities. Field research was conducted in the Bahcesaray district in the province of Van in Eastern Anatolia, Turkey. Thirty-two married women with children participated in in-depth interviews. Results indicate that childbirth is regarded as a natural duty of women. Women generally have little knowledge about their own bodies and reproductive health and have few avenues for obtaining information about health services. In several cases, women reported having been married as children. Because of their ignorance about sexuality and reproductive health issues, feelings of helplessness and anxiety were high. Women felt hemmed in by the pressure of normative agents such as their husbands, their mothers-in law and older members of the family, and they felt obliged to show respect to these authority figures. Findings demonstrate how in this context patriarchy directly controls women's lives and health in physiological terms. PMID- 18432427 TI - Sexual responses to living with HIV/AIDS in Lome, Togo. AB - A qualitative study of sexual behaviour among 88 people living with HIV was conducted in Lome, Togo. Data were collected by means of open-ended interviews. Three separate sexual-behaviour patterns were identified: people who reported not having sex at all; people who reported using condoms consistently; and people who reported not using condoms at all or using them inconsistently. Reasons given to explain these behaviours were analysed. Findings reveal the need for different programmes and policy approaches to dealing with problems of HIV and AIDS in Lome, Togo. PMID- 18432428 TI - HIV prevention and low-income Chilean women: machismo, marianismo and HIV misconceptions. AB - Socio-cultural factors and HIV-related misinformation contribute to the increasing number of Chilean women living with HIV. In spite of this, and to date, few culturally specific prevention activities have been developed for this population. The goal of the present study was to elicit the perspectives of low income Chilean women regarding HIV and relevant socio-cultural factors, as a forerunner to the development of a culturally appropriate intervention. As part of a mixed-methods study, fifty low-income Chilean women participated in a survey and twenty were selected to participate in prevention, in-depth interviews. Results show evidence of widespread misinformation and misconceptions related to HIV/AIDS. Machismo and marianismo offer major barriers to prevention programme development. Future HIV prevention should stress partner communication, empowerment and improving the education of women vulnerable to HIV. PMID- 18432431 TI - Approach to scoliosis changed due to causes other than evidence: patients call for conservative (rehabilitation) experts to join in team orthopedic surgeons. AB - PURPOSE: To look critically at the present reality of AIS (Adolescent Idiopathic Scoliosis) treatment and verify the hypothesis that the current prevalence of a single medical specialty could be creating distortions in patient care and/or cure. METHOD: This is a multifaceted study comprising a review of the evidence on AIS, a bibliometric study of the general and orthopedic literature since Medline start, and two case reports. RESULTS: Evidence exists to support the efficacy of exercises, bracing and fusion (grade B, B and C recommendations, respectively), but in clinics exercises are generally ignored; braces are used with some criticism, while fusion is generally considered the only reliable treatment. The literature on AIS treatment prevails in journals of orthopedic surgery, and therapy papers focused on surgery have increased from 34 to 55% over the past two decades. The two clinical cases show how an incorrect psychological approach to the patient and family, as well as inappropriate conservative treatments can have disastrous consequences for patients. CONCLUSIONS: Our results seem to confirm the initial hypothesis: The interest of the AIS treatment community (composed almost exclusively by orthopedic surgeons) has shifted toward fusion whereas research has increased, while conservative treatment is suffering a decrease in professional interest (and diminished research). AIS requires expert, committed evidence-based care, but other specialists totally devoted to conservative treatment, particularly (but not exclusively) Physical and Rehabilitation Medicine specialists, should enter the field to create better treating teams. PMID- 18432432 TI - Evaluation of scoliosis today: examination, X-rays and beyond. AB - PURPOSE: The purpose of this paper is to provide an overview of the techniques of evaluation of patients, suffering from idiopathic scoliosis. METHODS: The presentation is provided, concerning the medical history, clinical examination, conventional radiography, stereo-radiography, surface topography, ultrasounds, computer tomography, and magnetic resonance imaging, focusing on the points specific for the pathology of idiopathic scoliosis. RESULTS: Use of the scoliometer became systematic in the clinical evaluation. Quality of life questionnaires, including those endorsed by the Society on Scoliosis Orthopaedic and Rehabilitation Treatment (SOSORT), oriented towards scoliotic patients, gain on popularity and are extremely helpful to objectively evaluate the disability related to scoliosis. Classical radiography serves as the basic exam to determine the curve type and magnitude. Ultrasounds, computer tomography and magnetic resonance imaging are indicated in precisely defined clinical situations. Stereo radiography and surface topography seem to be the most promising techniques, however requiring standardisation. CONCLUSIONS: Apart from sophisticated measurements, the experience of a physician cannot be underestimated. High standard clinical evaluation will probably continue to serve as a reference for other methods of assessment of patients with scoliosis. Stereo-radiography and surface topography deserve common use, after standardization is achieved. PMID- 18432433 TI - What a school screening program could contribute in clinical research of idiopathic scoliosis aetiology. AB - PURPOSE: Scoliosis school screening (SSS) programs have clinically benefited many children through early detection and treatment, as it is clearly stated in the Consensus Paper which has been published by the Society on Scoliosis Orthopaedic and Rehabilitation Treatment (SOSORT). It is also through these SSS programs that almost all of our current knowledge on the natural history and curve progression of IS has been made known. The present study summarizes the contribution of SSS in research of IS aetiology. METHOD: We reviewed all the reports in relation to research of IS aetiology, which were published in peer-review journals and were originated from the Thriasio SSS program. RESULTS: Analysis comprises of reports for (i) IS prevalence, (ii) aetiological studies originated from school screening referrals which implicate numerous environmental and biological factors in IS pathogenesis, (iii) suggestions for a more efficient screening, (iv) IS natural history, and (v) the evolving aim of SSS based on the reported research. CONCLUSIONS: SSS should be adopted by policy makers, because its scope goes beyond the identification of IS at an early stage, contributing significantly into the research for IS aetiology. PMID- 18432434 TI - Neurophysiological basis of rehabilitation of adolescent idiopathic scoliosis. AB - BACKGROUND: Knowledge on mechanisms of neurophysiological control of trunk movement and posture could help in the development of rehabilitation programs and brace treatment in adolescent idiopathic scoliosis (AIS). AIMS: Reviewing up-to date research on neurophysiology of movement and posture control with the aim of providing basis for new researches in the field of AIS rehabilitation and background understanding for clinicians engaged in management of AIS. METHODS: Review of literature. RESULTS: We considered several neurophysiological issues relevant for AIS rehabilitation, namely, the peculiar organization of patterns of trunk muscle recruitment, the structure of the neural hardware subserving axial and arm muscle control, and the relevance of cognitive systems allowing mapping of spatial coordinates and building of body schema. DISCUSSION AND CONCLUSION: We made clear the reason why trunk control is generally carried out by means of very fast, feedforward or feedback driven patterns of muscle activation which are deeply rooted in our neural control system and very difficult to modify by training. We hypothesized that augmented sensory feedback and strength exercises could be an important stage in a rehabilitation program aimed at hindering, or possibly reversing, scoliosis progression. In this context we considered bracing not only as a corrective biomechanical device but also as a tool for continuous sensory stimulation that could help awareness of body misalignment. Future research aimed at developing strategies of trunk postural control learning is essential in the rehabilitation of adolescent idiopathic scoliosis. PMID- 18432435 TI - Exercises reduce the progression rate of adolescent idiopathic scoliosis: results of a comprehensive systematic review of the literature. AB - BACKGROUND: A previously published systematic review (Ped.Rehab.2003 - DARE 2004) documented the existence of the evidence of level 2a (Oxford EBM Centre) on the efficacy of specific exercises to reduce the progression of AIS (Adolescent Idiopathic Scoliosis). AIM: To confirm whether the indication for treatment with specific exercises for AIS has changed in recent years. STUDY DESIGN: Systematic review. METHODS: A bibliographic search with strict inclusion criteria (patients treated exclusively with exercises, outcome Cobb degrees, all study designs) was performed on the main electronic databases and through extensive manual searching. We retrieved 19 studies, including one RCT and eight controlled studies; 12 studies were prospective. A methodological and clinical evaluation was performed. RESULTS: The 19 papers considered included 1654 treated patients and 688 controls. The highest-quality study (RCT) compared two groups of 40 patients, showing an improvement of curvature in all treated patients after six months. We found three papers on Scoliosis Intensive Rehabilitation (Schroth), five on extrinsic autocorrection-based methods (Schroth, side-shift), four on intrinsic autocorrection-based approaches (Lyon and SEAS) and five with no autocorrection (three asymmetric, two symmetric exercises). Apart from one (no autocorrection, symmetric exercises, very low methodological quality), all studies confirmed the efficacy of exercises in reducing the progression rate (mainly in early puberty) and/or improving the Cobb angles (around the end of growth). Exercises were also shown to be effective in reducing brace prescription. CONCLUSION: In five years, eight more papers have been published to the indexed literature coming from throughout the world (Asia, the US, Eastern Europe) and proving that interest in exercises is not exclusive to Western Europe. This systematic review confirms and strengthens the previous ones. The actual evidence on exercises for AIS is of level 1b. PMID- 18432436 TI - Bracing adolescent idiopathic scoliosis: a systematic review of the literature of effective conservative treatment looking for end results 5 years after weaning. AB - PURPOSE: The purpose of this study is to investigate long-term radiological and clinical outcomes of brace treatment, comparing to those of the natural history or surgical treatment. METHOD: Review of literature. RESULTS: Most of the ling term follow-up study of brace treatment showed initial improvement with use of the brace, slight improvement compared with the original curve at the time of the brace discontinuation, and an additional loss of correction at the time of the follow-up. The rate of loss of correction was comparable to the progression rate observed in the natural history after skeletal maturity, and the effect of the brace treatment remained even with some loss of correction after the treatment. Clinical outcomes investigated were pain, function, and activity of the patients, including marital status, child bearing, and job performance. Most of the studies concluded that quality of life of the patients treated by brace was comparable to that of the control, with some exception reported regarding back pain and some kind of activities. CONCLUSION: Brace treatment is effective even in the long term for the treatment of adolescent idiopathic scoliosis. Curve progression after the treatment was equivalent to that in the natural history. Quality of life of the patients treated by brace was comparable to that of the normal control. PMID- 18432437 TI - Conservative management of neuromuscular scoliosis: personal experience and review of literature. AB - PURPOSE: The principles of conservative management of neuromuscular scoliosis in childhood and adolescence are presented. METHODS: Analysis of personal experience and literature review. The topic is discussed separately for patients with flaccid or spastic paresis. RESULTS: These demonstrate that conservative management might be proposed for patients with neuromuscular scoliosis in many clinical situations. In spastic disorders, it maintains the symmetry around the hip joints. Bracing is technically difficult and often is not tolerated well by cerebral palsy children. In patients with flaccid paresis, the fitting and the use of brace is easier than in spastic patients. The flexibility of the spinal curvature is more important. Functional benefits of conservative management of neuromuscular scoliosis comprise stable sitting, easier use of upper limbs, discharge of the abdomen from the collapsing trunk, increased diaphragm excursion, and, not always, prevention of curve progression. CONCLUSIONS: Specific natural history and multiple medical problems associated with the disease make the treatment of children with neuromuscular scoliosis an extremely complex issue, best addressed when a team approach is applied. Continuously improving techniques of conservative management, comprising bracing and physiotherapy, together with correctly timed surgery incorporated in the process of rehabilitation, provide the optimal care for patients. PMID- 18432438 TI - Adolescent idiopathic scoliosis (AIS) - an indication for surgery? A systematic review of the literature. AB - PURPOSE: Historically, the treatment options for AIS, the most common form of scoliosis are: Exercises, in-patient rehabilitation, braces and surgery. While there is evidence in the form of prospective controlled studies that Scoliosis Intensive Rehabilitation (SIR) and braces can alter the natural history of the condition, there is no review on prospective controlled trials for surgical treatment. The aim of this review was to perform a systematic search of the Pub Med literature to reveal the evidence on scoliosis surgery. METHODS: A systematic review has been performed using the Pub Med database. Literature has been searched for the outcome parameter; 'rate of progression' and only prospective controlled studies that have considered the treatment versus the natural history have been included. RESULTS: No controlled study, not in the short, mid or long term, searched within the review, has been found to reveal evidence to support the hypothesis that the effects of surgery as a treatment option for AIS is superior to natural history. CONCLUSIONS: No evidence has been found in terms of prospective controlled studies to support surgical intervention from the medical point of view. In the light of the unknown long-term effects of surgery and in concluding on the lack of evidence already found that surgery might change the signs and symptoms of scoliosis, a randomized controlled trial (RCT) is long overdue. Until such a time that such evidence exists, there can be no medical indication for surgery. The indications for surgery are limited for cosmetic reasons in severe cases and only if the patient and the family agree with this. PMID- 18432439 TI - A century of spine surgery: what can patients expect? AB - PURPOSE: To evaluate the hypothesis that spinal fusion surgery is an effective method to address spinal deformity-associated clinical problems, including magnitude of curvature (Cobb angle), pulmonary dysfunction, and pain. METHOD: A systematic review was carried out using Science Citation Index (SCI) Expanded (1900 - present), Social Sciences Citation Index (1956 - present), Arts and Humanities Citation Index (1965 - present), Medline (1950 - present) and PubMed Central databases (1887 - present) to access information regarding efficacy of spine surgery in preventing or improving the health and function of patients diagnosed with scoliosis in adolescence. RESULTS: Since 1950, more than 12,600 articles on scoliosis have been published, and nearly 50% (5721) focus on methods, rationale, outcome, and complications of surgical intervention. Among these, 82 articles have documented outcome for groups of > or =10 patients, treated for adolescent idiopathic scoliosis, and followed for at least 2 years after treatment. These data provide an overview of the impact of spine surgery on scoliosis for 5780 patients as surgery methods and approaches have evolved. CONCLUSIONS: For most patients, a reduced magnitude of spinal curvature can be achieved through one or more spinal fusion surgeries. There is no evidence to support the premise that this result is correlated with improved pulmonary function or reduced pain. PMID- 18432440 TI - Secular changes in psychophysically determined maximum acceptable weights and forces over 20 years for male industrial workers. AB - The most frequent and expensive cause of compensable workplace injuries loss is manual material handling (MMH). In an attempt to minimise these losses, refinement of existing MMH guidelines is a component of redesigning high risk MMH jobs. In the development of the present MMH 1991 guidelines (Snook and Ciriello 1991), maximum acceptable weights (MAWs) and forces (MAFs) were derived from studies conducted in a 21 year time span before the above publication date. The question arises whether the present generation of workers have the same psychophysically determined weights and forces as those reflected in the guidelines. Therefore, the present study investigated whether secular changes had occurred in key MMH tasks in trials performed by present day local industrial workers. A total of 23 male industrial workers performed 20 variations of lifting, lowering, pushing, pulling and carrying tasks. A psychophysical methodology, identical to that of the authors' previous experiments, was used whereby the subjects were asked to select a workload they could sustain for 8 h 'without straining themselves or without becoming unusually tired, weakened, overheated or out of breath'. The results revealed that MAWs of lifting, lowering and carrying averaged 69% of the guideline values. MAFs of pushing and pulling showed less of a drop, averaging 82% and 94% respectively for initial and sustained forces. The results also indicated that the effects of the variables frequency, height, lifting vs. lowering, pushing vs. pulling were similar to earlier reported results, even though the absolute weights or forces were lower. It was concluded that consideration to change existing guidelines, reflecting this new psychophysical set point, may be appropriate if these significant performance decreases are confirmed in other locations, with greater subject numbers, and by other investigators. PMID- 18432441 TI - The impact of operating heavy equipment vehicles on lower back disorders. AB - Literature reviews examining the relationship between heavy equipment vehicle (HEV) operation and the development of musculoskeletal disorders have generally been qualitative in nature and have not employed an evidence-based assessment procedure. This research determines the extent to which whole-body vibration/shock and working postures are associated with lower back and neck disorders among HEV operators, while accounting for individual (i.e. age, gender, prior history of back or neck disorders) and occupational (i.e. material handling, climatic conditions, psychosocial factors) confounders. Published articles were obtained from a search of electronic databases and from bibliographies in the identified articles. A critical appraisal of these articles was conducted using an epidemiological appraisal instrument (Genaidy et al. 2007). The meta-analysis was conducted using statistical techniques employing fixed-effect and random-effect models. Eighteen articles reporting observational studies satisfied the inclusion criteria adopted for this research. The methodological qualities of the published studies ranged from marginal to average. The meta-relative risk was found to be 2.21, indicating that operators exposed to driving HEVs are at more than twice the risk of developing lower back pain in comparison to those not exposed to driving HEVs. Therefore, it seems possible that there is a causal relationship between working as a HEV operator and development of lower back disorders. Prospective cohort studies are urgently needed to confirm the outcomes of this evidence-based methodology (based in part on the meta-analysis) and the biological plausibility should be further explored. The reported findings point to a need for improved ergonomic design of HEVs. PMID- 18432442 TI - Does musculoskeletal discomfort at work predict future musculoskeletal pain? AB - The objective of this prospective cohort study was to evaluate if peak or cumulative musculoskeletal discomfort may predict future low-back, neck or shoulder pain among symptom-free workers. At baseline, discomfort per body region was rated on a 10-point scale six times during a working day. Questionnaires on pain were sent out three times during follow-up. Peak discomfort was defined as a discomfort level of 2 at least once during a day; cumulative discomfort was defined as the sum of discomfort during the day. Reference workers reported a rating of zero at each measurement. Peak discomfort was a predictor of low-back pain (relative risk (RR) 1.79), neck pain (RR 2.56), right or left shoulder pain (RR 1.91 and 1.90). Cumulative discomfort predicted neck pain (RR 2.35), right or left shoulder pain (RR 2.45 and 1.64). These results suggest that both peak and cumulative discomfort could predict future musculoskeletal pain. PMID- 18432443 TI - A physical demands analysis of the 24-week British Army Parachute Regiment recruit training syllabus. AB - This study assessed the physical demands of the 24-week Combined Infantryman's Course (CIC) for Parachute Regiment (Para) recruits and developed physical selection standards for applicants. Fifty recruits were monitored over five separate periods (35 d in total during weeks 1-2, 5, 9, 15 and 19-20). Energy expenditure (doubly labelled water), physical activity (accelerometry) and cardiovascular strain (% heart rate reserve) were measured. There was no overall progression in both the levels of physiological stress (physical activity counts and energy expenditure) and resultant cardiovascular strain during the first nine weeks of training. Applicants' 2.4 km run time and static lift strength measured at selection predicted 10 mile loaded march performance at the end of CIC Para. The introduction of job-related selection procedures and a more progressive approach to training has reduced the incidence of medical discharge from 14.4% to 5.1% and increased overall pass rates from 43% to 58%. PMID- 18432444 TI - Estimation of energy expenditure in a work environment: comparison of accelerometry and oxygen consumption/heart rate regression. AB - The aim of this study was to compare estimation of energy expenditure (EE) in working environments, either from accelerometry or from an individual oxygen consumption/heart rate (VO(2)/HR) regression curve. The study participants were 46 volunteer workers aged 27+/-6 years old. A significant correlation between EE predicted by the VO(2)/HR curve and the accelerometer was observed (r=0.78, p <0.01). However, more disparities were observed between the two methods when the mean job intensity was not within 16% and 23% higher than resting HR. The accelerometer overestimated by a mean of 34.4% the prediction by VO(2)/HR regression if the intensity of the task was lower than a total of 1000 kcal/shift and underestimated the prediction by a mean of -24.9% if EE estimation of the work shift was higher than a total of 1500 kcal/shift. Despite a high correlation between both methods in the whole group, EE evaluated by accelerometry does not correspond to EE predicted by the VO(2)/HR regression curves when evaluated individually. PMID- 18432445 TI - Evaluation of proper height for squatting stool. AB - Many jobs and activities in people's daily lives have them in squatting postures. Jobs such as housekeeping, farming and welding require various squatting activities. It is speculated that prolonged squatting without any type of supporting stool would gradually and eventually impose musculoskeletal injuries on workers. This study aims to examine the proper height of the stool according to the position of working materials for the squatting worker. A total of 40 male and female college students and 10 female farmers participated in the experiment to find the proper stool height. Student participants were asked to sit and work in three different positions: floor level of 50 mm; ankle level of 200 mm; and knee level of 400 mm. They were then provided with stools of various heights and asked to maintain a squatting work posture. For each working position, they were asked to write down their thoughts on a preferred stool height. A Likert summated rating method as well as pairwise ranking test was applied to evaluate user preference for provided stools under conditions of different working positions. Under a similar experimental procedure, female farmers were asked to indicate their body part discomfort (BPD) on a body chart before and after performing the work. Statistical analysis showed that comparable results were found from both evaluation measures. When working position is below 50 mm, the proper stool height is 100 or should not be higher than 150 mm. When working position is 200 mm, the proper stool height is 150 mm. When working position is 400 mm, the proper stool height is 200 mm. Thus, it is strongly recommended to use proper height of stools with corresponding working position. Moreover, a wearable chair prototype was designed so that workers in a squatting posture do not have to carry and move the stool from one place to another. This stool should ultimately help to relieve physical stress and hence promote the health of squatting workers. This study sought to evaluate and make suggestions on user preference and BPD of proper stool height, at which work can be done comfortably, according to squatting work position. In short, results showed that proper stool height differed according to working position, even with similar squatting tasks. PMID- 18432446 TI - The effects of dynamic movement on seated reach arcs. AB - The objective of this study was to determine the relationship between movement of the low back and shoulder during a normal seated reach and the reach arc estimation equations found in literature. The method consisted of evaluating individuals who were reaching with their right hands for five chess pawns, which were placed at varying distances. Specifically, the pawns were evenly spaced in a straight line directly in front of each participant's shoulder. This study focused on a group of 32 participants, which included both males and females. For each participant, low back, elbow and two shoulder angles were collected using a PEAK motion capture 6.0 system. Angles were collected in both the sagittal and transverse planes to gain a 3-D perspective. Data were summarised and correlated against maximum reach arc estimates. Results from the data suggest that both the shoulder and low back are engaged much earlier in a person's reach cycle than previously believed. Specifically, the results show low back engagement (trunk/lumber flexion) as early as 50% of maximum reach with the angle increasing to 5 degrees at around 80% and 10 degrees at 93%, which allows more pronounced forward angular acceleration. This shifts the shape and effective area of a participant's reach arc to a 'dynamic' state and questions if major muscle recruitment in the torso has initiated. The resulting effect is that dynamic and static reach arcs may vary significantly. While this study is too limited to support formal conclusions, these results strongly suggest a need for further investigation into the limits and impact of dynamic reach. PMID- 18432447 TI - Bone conduction reception: head sensitivity mapping. AB - This study sought to identify skull locations that are highly sensitive to bone conduction (BC) auditory signal reception and could be used in the design of military radio communication headsets. In Experiment 1, pure tone signals were transmitted via BC to 11 skull locations of 14 volunteers seated in a quiet environment. In Experiment 2, the same signals were transmitted via BC to nine skull locations of 12 volunteers seated in an environment with 60 decibels of white background noise. Hearing threshold levels for each signal per location were measured. In the quiet condition, the condyle had the lowest mean threshold for all signals followed by the jaw angle, mastoid and vertex. In the white noise condition, the condyle also had the lowest mean threshold followed by the mastoid, vertex and temple. Overall results of both experiments were very similar and implicated the condyle as the most effective location. PMID- 18432448 TI - The impact of background speech varying in intelligibility: effects on cognitive performance and perceived disturbance. AB - Noise abatement in office environments often focuses on the reduction of background speech intelligibility and noise level, as attainable with frequency specific insulation. However, only limited empirical evidence exists regarding the effects of reducing speech intelligibility on cognitive performance and subjectively perceived disturbance. Three experiments tested the impact of low background speech (35 dB(A)) of both good and poor intelligibility, in comparison to silence and highly intelligible speech not lowered in level (55 dB(A)). The disturbance impact of the latter speech condition on verbal short-term memory (n=20) and mental arithmetic (n=24) was significantly reduced during soft and poorly intelligible speech, but not during soft and highly intelligible speech. No effect of background speech on verbal-logical reasoning performance (n=28) was found. Subjective disturbance ratings, however, were consistent over all three experiments with, for example, soft and poorly intelligible speech rated as the least disturbing speech condition but still disturbing in comparison to silence. It is concluded, therefore, that a combination of objective performance tests and subjective ratings is desirable for the comprehensive evaluation of acoustic office environments and their alterations. PMID- 18432449 TI - The synergic role of sociotechnical and personal characteristics on work injuries in mines. AB - Occupational injuries in mines are attributed to many factors. In this study, an attempt was made to identify the various factors related to work injuries in mines and to estimate their effects on work injuries to mine workers. An accident path model was developed to estimate the pattern and strength of relationships amongst the personal and sociotechnical variables in accident/injury occurrences. The input data for the model were the correlation matrix of 18 variables, which were collected from the case study mines. The case study results showed that there are sequential interactions amongst the sociotechnical and personal factors leading to accidents/injuries in mines. Amongst the latent endogenous constructs, job dissatisfaction and safe work behaviour show a significant positive and negative direct relationship with work injury, respectively. However, the construct safety environment has a significant negative indirect relationship with work injury. The safety environment is negatively affected by work hazards and positively affected by social support. The safety environment also shows a significant negative relationship with job stress and job dissatisfaction. However, negative personality has no significant direct or indirect effect on work injury, but it has a significant negative relationship with safe work behaviour. The endogenous construct negative personality is positively influenced by job stress and negatively influenced by social support. PMID- 18432455 TI - The protective effect of Nigella sativa on lung injury of sulfur mustard-exposed Guinea pigs. AB - The lung is one of the most exposable organs to chemical warfare agents such as sulfur mustard (SM) gas. Airway hyperresponsiveness and lung inflammation are reported in chemical warfare victims. There is no definite treatment for respiratory disorders induced by SM exposure. However, the protective effect of Nigella sativa on inflammatory process was shown. In the present study, the protective effect of Nigella sativa on tracheal responsiveness and lung inflammation of SM exposed guinea pigs was examined. Guinea pigs were exposed to diluent's solution (ethanol, control group), 100 mg/m(3) inhaled sulfur mustard (SME group), and SME treated with Nigella sativa, 0.08 g daily (SME+N), n = 6 for each group. Tracheal responsiveness (TR) to methacholine, total white blood cell (WBC) count of lung lavage, and differential WBC were done 14 days post exposure. The weigh of animal were measured at the beginning, middle (day 7), and the end (day 14) of the study. The TR of SM-exposed guinea pigs was significantly (P < .001) and WBC nonsignificantly higher than those of controls. In SME guinea pigs, there was a weight loss but in the case of SME+N guinea pigs, no obvious weight change thought the study was seen. The eosinophl, monocyte, and lympocytes in SME animals were significantly changed compared to control group (P < .001 for all cases). Monocyte, lymphocyte, and neutrophil number were decreased in SME+N group compared to SME animals, which was significant only for neutrophil (P < .05). These results showed a preventive effect of Nigella sativa on TR of SM-exposed guinea pigs. PMID- 18432454 TI - The calcitriol analogue EB1089 impairs alveolarization and induces localized regions of increased fibroblast density in neonatal rat lung. AB - The active form of vitamin D3, 1,25-dihydroxyvitamin D3 (1,25-(OH)2D3, or calcitriol), is a potent mitogen for fibroblasts cultured from rat lungs at postnatal day 4 (P4), during the peak of septation (P3 to P7). In light of the key role of fibroblasts in alveolar septation, the authors conducted studies to measure the extent to which 1,25-(OH)2D3 affects lung maturation in vivo, as well as its ability to influence the stimulatory activity of all-trans retinoic acid (RA). To identify a calcitriol analogue with maximal mitogenic activity and low systemic toxicity, two compounds with reduced calcemic activity (EB1089 and CB1093) and a superagonist (MC1288) were evaluated in neonatal rat lung fibroblast cultures. All 3 analogues were more potent mitogens than 1,25 (OH)(2)D3 itself (MC1288 approximately CB1093 > EB1089 > 1,25-(OH)2D3). In addition, each was more effective than 1,25-(OH)2D3(EB1089 > CB1093 > MC1288 > 1,25-(OH)2D3) in the activation of a vitamin D response element from the platelet derived growth factor (PDGF)-A gene, whose expression is essential for normal alveolarization. Daily administration of EB1089 to rats 4 to 12 days of age caused an increase in mean alveolar chord length (P < .0001), and also elicited prominent regions of fibroblast hypercellularity, as defined in terms of a vimentin-positive, factor VIII-negative phenotype. EB1089 and RA each induced the expression of 2 important lung structural proteins, collagen and elastin. Regions of fibroblast hypercellularity induced by EB1089 were strongly positive for expression of the alveolarization-relevant growth factors, PDGF-AA and vascular endothelial growth factor (VEGF). These studies demonstrate that 1,25-(OH)2D3 disrupts the overall alveolarization process in the neonatal lung, although it stimulates expression of some proteins associated with lung morphogenesis. PMID- 18432456 TI - Regulation of pulmonary vasoconstriction by agonists and caveolae. AB - Sustained pulmonary vasoconstriction contributes to the elevated pulmonary vascular resistance observed in pulmonary arterial hypertension. A rise in cytosolic Ca(2 +) in pulmonary artery smooth muscle cells (PASMCs) is major trigger for pulmonary vasoconstriction. One family of drugs currently being pursued as a potential treatment for pulmonary hypertension are the statins, which act by depleting cholesterol and reducing the number of caveolae. This study aimed at investigating the role of caveolae, membrane receptors, and ion channels (that are potentially located in the caveolae) in agonist-mediated pulmonary vasoconstriction in order to gain a greater understanding of the signaling mechanisms involved in the regulation of pulmonary vascular tone. Chronic treatment of PASMCs with the cholesterol-depleting agent, methyl-beta cyclodextrin (Mbeta CD), significantly reduced the number of cholesterol rich caveolae regions in the membrane. This disruption of cholesterol in caveolae significantly inhibited pharmacomechanical (induced by phenylephrine), but not electromechanical (induced by elevated extracellular potassium concentration), rat pulmonary artery contraction. These results indicate that receptors may functionally colocalize in caveolae in PASMCs and coordinate to regulate pulmonary vascular tone. PMID- 18432457 TI - Hearing the voices of people with communication disabilities. PMID- 18432458 TI - Exposed and embedded corrections in aphasia therapy: issues of voice and identity. AB - BACKGROUND: Because communication after the onset of aphasia can be fraught with errors, therapist corrections are pervasive in therapy for aphasia. Although corrections are designed to improve the accuracy of communication, some corrections can have social and emotional consequences during interactions. That is, exposure of errors can potentially silence the 'voice' of a speaker by orienting to an utterance as unacceptable. Although corrections can marginalize speakers with aphasia, the practice has not been widely investigated. AIMS: A qualitative study of corrections during aphasia therapy was undertaken to describe corrections in therapy, identify patterns of occurrence, and develop hypotheses regarding the potential effects of corrections. METHODS & PROCEDURES: Videotapes of six individual and five group aphasia therapy sessions were analysed. Sequences consistent with a definition of a therapist 'correction' were identified. Corrections were defined as instances when the therapist offered a 'fix' for a perceived error in the client's talk even though the intent was apparent. OUTCOMES & RESULTS: Two categories of correction were identified and were consistent with Jefferson's (1987) descriptions of exposed and embedded corrections. Exposed corrections involved explicit correcting by the therapist, while embedded corrections occurred implicitly within the ongoing talk. Patterns of occurrence appeared consistent with philosophical orientations of therapy sessions. Exposed corrections were more prevalent in sessions focusing on repairing deficits, while embedded corrections were prevalent in sessions focusing on natural communication events (e.g. conversation). In addition, exposed corrections were sometimes used when client offerings were plausible or appropriate, but were inconsistent with therapist expectations. CONCLUSIONS: The observation that some instances of exposed corrections effectively silenced the voice or self-expression of the person with aphasia has significant implications for outcomes from aphasia therapy. By focusing on accurate productions versus communicative intents, therapy runs the risk of reducing self-esteem and communicative confidence, as well as reinforcing a sense of 'helplessness' and disempowerment among people with aphasia. The results suggest that clinicians should carefully calibrate the use of exposed and embedded corrections to balance linguistic and psychosocial goals. PMID- 18432459 TI - Voices of disability on the radio. AB - BACKGROUND: While much commentary exists in relation to the portrayal of disabled people in the media, very little research examines the talk itself in any detail. This paper examines the how people with communication disabilities and disabled people are dealt with in the talk of a radio programme about disability. AIMS: To show how the voices of disabled people, and in particular people with communication difficulties, are dealt with on a radio programme titled For and About People with Disability (http://www.rte.ie). METHODS & PROCEDURES: Analysis of 15 episodes of an Irish radio programme for and about people with disability called Outside the Box to identify frames governing the discourse. OUTCOMES & RESULTS: Three frames are identified: radio programme frame; presenter frame; and interview frame. Communication disability never appears as a topic in the radio programme frame. In the presenter and interview frames the presenter foregrounds medical aspects of experiences, asks questions that only seek factual information, and fails to respond to subjective aspects of disabled people's experiences. Analysis of the interviewees' responses show how they hold the floor and introduce subjective accounts of living with disability. CONCLUSIONS: Frame analysis reveals how disability (including communication disability) is dealt with in the talk of a radio programme for and about people with disability. There is an overemphasis on medical aspects of disability and a view of disability as a primarily physical phenomenon is broadcast. In spite of mainly medical/factual questions, interviewees manage to include rich accounts of their experience. People with communication disability are not included, possibly due to issues of intelligibility or lack of awareness. Speech and language therapists have a valuable role to play in terms of 'learning to listen' and 'helping to tell'. PMID- 18432460 TI - Listening to the voice of living life with aphasia: Anne's story. AB - BACKGROUND: Listening to how people talk about the consequences of acquired aphasia helps one gain insight into how people construe disability and communication disability in particular. It has been found that some of these construals can be more of a disabling barrier in re-engaging with life than the communication impairment itself. AIMS: To explore and identify the role that narratives of disability (i.e. the 'inner stories we live by') play in how Anne, a woman with aphasia, and those close to her, make sense of stroke and aphasia. METHODS & PROCEDURES: Qualitative analyses of in-depth interviews, of notes from participant observation, and of artefactual material were used to get access to these insider perspectives of both Anne and some of her conversation partners. OUTCOMES & RESULTS: The data revealed that Anne created, was surrounded by, and immersed in an intricate and elusive tapestry of narratives of disability that contributed to her experience of living life with aphasia. Narratives of 'limited competence' and 'disability as less than whole' emerged as strong voices that stemmed from cultural and societal views about disability. In response to this Anne and other participants sought refuge in the 'grand' narrative of Modern Medicine with its focus on restitution, in order to make sense of their situation and to chart a way forward. As a result her life revolved around activities that would 'make her better'. CONCLUSIONS: Prevailing cultural narratives of disability played a key role in how Anne and those close to her made sense of aphasia and so shaped how she lived her life. The implications of this for working practice are discussed. PMID- 18432461 TI - Representing voices from the life-world in evidence-based practice. AB - BACKGROUND: Current models of evidence-based practice marginalize and even silence the voices of those who are the potential beneficiaries of assessment and intervention. These missing voices can be found in the reflections of clients on their own life-world experiences. AIMS: This paper examines how voices from the life-world are silenced in current models of evidence-based practice and why this is inappropriate. It is argued that personal experience narratives rooted in the inter-subjective life-worlds of clients - currently conceptualized as mere anecdotal information or consumer opinion in evidence-based practice - should be elevated to the status of a bona fide unit of analysis in the construction of evidence. The aim is not to supplant more traditional, objective units of measurement, but, rather, to enhance our ability to interpret the ecological validity of evidence by listening to our clients' voices. METHODS & PROCEDURES: Written discourse from professional position statements, technical reports, and other sources of information are analysed in terms of how they conceptualize evidence within evidence-based practice. Findings from this analysis are then compared with personal experience narratives from clients, and significant others, reflecting on the outcomes of assessment and intervention practices. OUTCOMES & RESULTS: Analysis reveals that current quality of evidence hierarchies in evidence-based practice are based on an epistemology of logical positivism. They privilege cause-effect explanations grounded in objective, empirically testable variables that exist external to the self. Unfortunately, the dismissal of subjective, phenomenally oriented information has functioned to marginalize and silence voices from the life-worlds of clients when constituting proof of effectiveness. When these voices are excluded, our understanding of the social significance, magnitude and ecological validity of evidence is compromised. The current version of EBP needs to be reformulated to include subjective voices from the life-worlds of clients as a form of evidence. Toward this end, the personal experience narrative is proposed as one potential qualitative unit of analysis for characterizing the life-world outcomes of assessment and intervention. These narratives seem well suited to the task of representing social outcomes because of their attention to the particularities and consequences of human experience, and because of the way time is represented retrospectively. CONCLUSIONS: To understand the complex nature of communication disorders and the efficacy of our helping practices requires not only the analysis of objective material phenomena like part-word repetitions, phonemes and morphemes, but also an examination of how these phenomena are interpreted and valued as forms of communication by clients and significant others in the life-world. Personal experience narrative is suggested as one means for understanding experienced outcomes. Other qualitatively oriented methodologies and concepts associated with ethnography of communication and discourse analysis will undoubtedly be useful in this regard. PMID- 18432462 TI - Multiple voices in clinical discourse and as clinical intervention. AB - BACKGROUND: Grounded in sociocultural theories of language development and use, this paper explores the concept of dialogic voice. Building on the term 'dialogue', dialogic voice points to the fundamentally social nature of language in-use. From this perspective, language emerges from specific histories and thus carries the multiple voices of previous speakers. People draw on these voices to think about and represent the world, communicate with others, construct their own identities, and engage in play. Prior (2001) identified three key dimensions of dialogic voicing: typified social voices, re-envoicing others' words and acts, and personalized voice. AIMS: To present a theoretical framing for dialogic voice; to detail the dimensions of dialogic voice; and to offer a preliminary analysis of dialogic voicing in clinical discourse. METHODS & PROCEDURES: Data consisted of ten treatment sessions for a 67-year-old man with amnesia and aphasia, using a collaborative barrier task protocol. Discourse analysis and selective coding were used to identify the three dimensions of dialogic voice in both clinician and client utterances. OUTCOMES & RESULTS: During this collaborative task, the client and clinician produced an array of voices, including: (1) typified social voices to display professional expertise, family identities, and shared interests; (2) re-envoicing others' words and acts in both task and non-task interactions; and (3) personalized voicing, displayed mostly in the client's discourse. CONCLUSIONS: Attention to dialogic voicing offers a way to see and reflect on the heterogeneity of discourse and the multiple identities that clinicians and clients alike can, and do, display in clinical settings. Tracing the complex interplay of multiple voices provides us with insights into rich communicative environments that, from a sociocultural perspective, provide opportunities for initiating change in the communicative practices of clients, their communicative partners, and ultimately clinical practice itself. PMID- 18432463 TI - Multiple voices in Charles Van Riper's desensitization therapy. AB - BACKGROUND: Charles Van Riper (1905-94) author, researcher and practitioner, was one of the major contributors to the field of stuttering in the 20th century. His series of Action Therapy videotapes provide a useful model of how therapy was implemented by a master clinician. AIMS: Van Riper's session with a client is analysed to reveal how voices are used by these two participants during desensitization therapy, and to demonstrate how Van Riper's expertise manifests itself during the session. METHODS & PROCEDURES: Interaction between Van Riper and a client during the desensitization stage of therapy was transcribed, annotated, and analysed using Goffman's (1974) frame analysis. Elements in the extensive discourse (about 4800 words exchanged) were chosen to illustrate a range of distinctive voices used by Van Riper and the client during this 'crucial' stage of therapy. OUTCOMES & RESULTS: Van Riper shifts his voice throughout the session. His comments are sometimes directed to the viewing audience, at which times he tends to use the voice of an exegeter. When directing his comments to the client, he uses two voices: an authoritative one and one that aligns with the person's feelings. The voice of the client is often silenced, especially when Van Riper is being authoritative. Sometimes Van Riper uses his expertise by speaking through the voice of the client. This is especially true when summarizing what the client might be feeling. The true voice of the client emerges only occasionally. Detailed discourse analysis focusing on the roles played by participants as revealed through footings taken by speakers show complexity in the interaction that is not obvious at surface level. CONCLUSIONS: By using discourse analysis, the multiple voices that Van Riper uses to achieve the complex tasks involved in desensitization can be specified. PMID- 18432464 TI - Whose voice is it anyway? Hushing and hearing 'voices' in speech and language therapy interactions with people with chronic schizophrenia. AB - BACKGROUND: Some people with schizophrenia are considered to have communication difficulties because of concomitant language impairment and/or because of suppressed or 'unusual' communication skills due to the often-chronic nature and manifestation of the illness process. Conversations with a person with schizophrenia pose many pragmatic challenges as they may include delusional talk or hallucinatory references considered irrelevant or inappropriate to ongoing conversation. Yet delusional talk and references to hallucinations are often an intrinsic part of the overall presentation in people with schizophrenia. The speech and language therapist (SLT) must be aware of the influence and function of these pragmatic challenges in therapeutic interactions with this client group. One way of increasing this awareness in this context is through an exploration of co-constructed 'voices' in speech and language therapy (SLT) interactions. AIMS: This paper aims to explore the 'voices' in speech and language therapy interactions with clients with schizophrenia. Identifying and revealing voices as constructed or negotiated by participants should help to illuminate the construction of meaning and the nature of the therapeutic interaction in this context. MAIN CONTRIBUTION: The analysis of conversations between people with schizophrenia, a speech and language therapist, and speech and language therapy students during clinical sessions reveals 'voices' being aired in the interactions. These voices, described primarily as 'the voice of speech and language therapy' and 'the voice of the lived experience of schizophrenia', are jointly constructed and 'heard' to greater or lesser extents depending on the context of the interaction. The analysis illustrates that whereas the speech and language therapist's primary agenda is actively to engage the client in conversational interaction for assessment and therapeutic purposes (i.e. the voice of speech and language therapy), this is sometimes (but not always) at the cost of hushing other talk. This is especially the case when the talk is delusional and seemingly inappropriate within the clinician-defined agenda. However, talk about the nature of the illness (and in particular hallucinations) and the potential impact on communication is compatible with the clinician's agenda and allowed to proceed. Therefore, the voice of the lived experience of schizophrenia is silenced within delusional talk, yet heard in talk about hallucinatory experiences and communication difficulties. It will be shown how these voices interact to illuminate the participants' agendas and define the therapeutic interaction in this context; that is to say, the therapist manipulates talk that fails to conform to specific speech and language therapy goals, preferring to promote and encourage agenda-relevant talk. CONCLUSIONS: The analysis shows that the voice of SLT and the voice of the lived experience of schizophrenia are heard to greater or lesser extents, as determined by the context and content of the talk-in-interaction. However, the voice of SLT must blend with, rather than hush, the plurality of voices that characterise therapeutic interactions. An increased awareness of competing voices, as they struggle to be heard in SLT interactions, may prompt therapists to consider and evaluate their often agenda-driven influence on the unfolding interactions we call 'therapy'. PMID- 18432465 TI - Intermittent pneumatic compression - systems and applications. AB - PRIMARY OBJECTIVE: The purpose of this review is to survey the types of intermittent pneumatic compression systems that are currently used, and their medical applications. MAIN OUTCOMES AND RESULTS: Intermittent compression devices have taken many forms since their initial development, but medical justifications for particular properties of cuff design, compression timing and pressure are often weak. Intermittent compression is well established, and effective in the prevention of deep vein thrombosis (DVT) and reduction of lymphoedema. Other therapeutic applications, such as in chronic arterial and venous disease, are not yet as well accepted, but may become more popular as published evidence increases. CONCLUSIONS: The full potential of intermittent pneumatic compression has probably not yet been realized, and requires better quality research. System design must follow physiological evidence, and while complexity in that design may allow greater therapeutic flexibility, it may incur greater financial cost, difficulty in use, and in the prevention of DVT in particular may be unnecessary. PMID- 18432466 TI - Imaging of breast cancer with mid- and long-wave infrared camera. AB - In this novel study the breasts of 15 women with palpable breast cancer were preoperatively imaged with three technically different infrared (IR) cameras - micro bolometer (MB), quantum well (QWIP) and photo voltaic (PV) - to compare their ability to differentiate breast cancer from normal tissue. The IR images were processed, the data for frequency analysis were collected from dynamic IR images by pixel-based analysis and from each image selectively windowed regional analysis was carried out, based on angiogenesis and nitric oxide production of cancer tissue causing vasomotor and cardiogenic frequency differences compared to normal tissue. Our results show that the GaAs QWIP camera and the InSb PV camera demonstrate the frequency difference between normal and cancerous breast tissue; the PV camera more clearly. With selected image processing operations more detailed frequency analyses could be applied to the suspicious area. The MB camera was not suitable for tissue differentiation, as the difference between noise and effective signal was unsatisfactory. PMID- 18432467 TI - A wavelet-based optimal texture feature set for classification of brain tumours. AB - In this work, the classification of brain tumours in magnetic resonance images is studied by using optimal texture features. These features are used to classify three sets of brain images - normal brain, benign tumour and malignant tumour. A wavelet-based texture feature set is derived from the region of interest. Each selected brain region of interest is characterized with both its energy and texture features extracted from the selected high frequency subband. An artificial neural network classifier is employed to evaluate the performance of these features. Feature selection is performed by a genetic algorithm. Principal component analysis and classical sequential methods are compared against the genetic approach in terms of the best recognition rate achieved and the optimal number of features. A classification performance of 98% is achieved in a genetic algorithm with only four of the available 29 features. Principal component analysis and classical sequential methods require a larger feature set to attain the similar classification accuracy of 98%. The optimal texture features such as range of angular second moment, range of sum variance, range of information measure of correlation II and energy selected by the genetic algorithm provide best classification performance with lower computational effort. PMID- 18432468 TI - Detection of QRS complexes in electrocardiogram using support vector machine. AB - This paper presents the application of a support vector machine (SVM) for the detection of QRS complexes in the electrocardiogram (ECG). The ECG signal is filtered using digital filtering techniques to remove noise and baseline wander. The support vector machine is used as a classifier to delineate QRS and non-QRS regions. Two different algorithms are presented for the detection of QRS complexes. The first uses a single-lead ECG at a time for the detection of QRS complexes, while the second uses 12-lead simultaneously recorded ECG. Both algorithms have been tested on the standard CSE ECG database. A detection rate of 99.3% is achieved when tested using a single-lead ECG. This improves to 99.75% for the simultaneously recorded 12-lead ECG signal. The percentage of false negative detection is 0.7% and the percentage of false positive detection is 12.4% in the single-lead QRS detection and it reduces to 0.26% and 1.61% respectively for QRS detection in simultaneously recorded 12-lead ECG signals. The performance of the algorithms depends strongly on the selection and the variety of the ECGs included in the training set, data representation and the mathematical basis of the classifier. PMID- 18432469 TI - Non-contact determination of arterial blood pressure alterations induced by blood loss using laser irradiation on the common carotid artery. AB - In order to avoid the secondary exposure of medical personnel to toxic materials under biochemical hazard conditions, we have reported a method for non-contact monitoring of heart and respiratory rates, using microwave radar or laser irradiation. In large-scale disasters, it is important to be able to diagnose shock without touching patients. We evaluated a non-contact method of monitoring arterial blood pressure alterations of New Zealand rabbits induced by blood loss, using He-Ne laser reflection on the common carotid artery. PVR was significantly correlated with systolic blood pressure (r = 0.95, p < 0.01), where PV = peak voltage of reflected laser amplitude, and PVR = PV(present moment state)/PV(normal state). The following formula was derived using the least squares linear fitting: SBP = 69.6 PVR + 8.2, in which SBP is the systolic blood pressure. Before blood withdrawal, the mean blood pressure, heart rate and haematocrit were 68 +/- 3 mmHg, 154 +/- 10 bpm and 40 +/- 2%, respectively. After intervention, the mean blood pressure, heart rate and haematocrit were 38 +/- 5 mmHg, 197 +/- 25 bpm and 30 +/- 2%, respectively. The proposed non-contact method appears promising for future clinical application in determining arterial blood pressure alterations. It is likely to be useful in reducing the risk of secondary exposure to toxic chemicals or infectious organisms in the case of large-scale disasters. PMID- 18432470 TI - Can an intermittent pneumatic compression system monitor venous filling in the leg? AB - OBJECTIVE: Intermittent pneumatic compression (IPC) systems are used for prophylaxis of venous thromboembolism. Both legs are wrapped with inflatable sleeves connected to a pneumatic controller to allow compression of the legs causing expulsion of venous blood. Venous refill between inflation periods causes leg expansion, which can be tracked by measuring pressure changes in the sleeve. The aim of our study, which utilized the SCD RESPONSE compression system in conjunction with an independent pressure transducer, was to investigate whether factors such as temperature changes within the sleeves during inflation and deflation affect the measured venous refill time (VRT). METHODS: Transducers were used to measure air pressure in the middle chamber of the sleeve. A thermocouple was also inserted into the bladder to measure temperature changes. Inflation, deflation and refill measurements were made with the sleeves around model systems (static, rigid plastic pipes or compliant paper rolls, and dynamic, latex tubes inserted between a rigid pipe and the sleeve to simulate veins) and on 10 subjects in semi-recumbent, supine and sitting positions. RESULTS: In all the experiments the maximum temperature change was 0.023 degrees C. With the static model systems, the pressure in the venous refill measuring bladder fell from the inflation pressure of 40 - 50 mmHg to 9 +/- 1 mmHg, but then rose by 2.1 +/- 0.2 mmHg (rigid pipes) and 1.4 +/- 0.2 mmHg (paper rolls). These pressure changes were associated with reported 'filling times' of 21 - 24 s (rigid pipes) and 22 - 29 s (paper rolls). In experiments on dynamic filling of the latex tube, there was a strong linear relationship between the filling time indicated by the SCD system and the time to empty the filling reservoir. In 170 measurements on human subjects, there were only three VRTs less than 30 s and 36 less than 35 s. VRT increased in all subjects when going from supine (34.6 +/- 1.8 s) to semi recumbent (38.9 +/- 1.9 s) to sitting (42.6 +/- 0.9 s) positions. DISCUSSION: In all cases, temperature changes during the refill phase were too small to result in significant pressure changes that would affect VRT. The pressure increases observed with the static models after deflation appeared to be due to viscoelastic relaxation. Viscoelastic responses were present in human subjects, but the effect on VRT was negligible. This indicates that the increased VRT observed in humans is due to blood return. Body position affected VRTs, indicating the system's ability to detect changes in filling times and venous blood volume. PMID- 18432471 TI - Iris recognition based on key image feature extraction. AB - In iris recognition, feature extraction can be influenced by factors such as illumination and contrast, and thus the features extracted may be unreliable, which can cause a high rate of false results in iris pattern recognition. In order to obtain stable features, an algorithm was proposed in this paper to extract key features of a pattern from multiple images. The proposed algorithm built an iris feature template by extracting key features and performed iris identity enrolment. Simulation results showed that the selected key features have high recognition accuracy on the CASIA Iris Set, where both contrast and illumination variance exist. PMID- 18432472 TI - A novel segmentation method using multiresolution analysis with 3D visualization for X-ray coronary angiogram images. AB - Coronary angiography is a widely used tool in the diagnosis and treatment of cardiac diseases. The main cause of coronary artery disease is atherosclerosis, which leads to the narrowing of artery lumen, resulting in decreased blood supply to heart muscles. Determination of narrowing of the lumens mainly depends upon the quality of the segmented image; with improved segmentation technique there is better accuracy in identification of blocks. The main purpose of the paper is to develop an automatic, accurate segmentation technique with 3D visualization for the segmented images. 3D visualization provides clearer information regarding the shape and severity of the lesion. The thresholding technique is one of the oldest and simplest techniques used for segmentation. This paper proposes a multithresholding approach using the entropy measure and multiresolution analysis to ensure automatic and accurate segmentation by overcoming some of the problems encountered in other techniques. Also, segmentation performance analysis was conducted for various segmentation methods. This method is tested with different real coronary angiographic images and was found to perform better than the other techniques. PMID- 18432473 TI - An investigation on pulse transit time in respiratory sleep studies for infants. AB - The main objective of this study was to assess the capability of pulse transit time (PTT) to identify obstructive and/or central apnoeas from normal breathing patterns in sleeping infants. Clinical trials were conducted with PTT being monitored as an adjunct measure in routine overnight polysomnography (PSG) studies. Standard PSG scorings of events were performed by two blinded observers to evaluate PTT accuracy. Data from eight infants (six male; aged 8.5 +/- 2.1 months; apnoea-hypopnoea index of 12.1 +/- 8.4 events per hour) were collected. Obstructive events were not successfully identified by PTT due to their low occurrence. However, PTT scored positive and negative predictive values of 0.803 and 0.788 for central events, respectively. Preliminary findings suggest that PTT has the potential to differentiate normal breathing from central events only in sleeping infants. PMID- 18432476 TI - Engineering case reports. Effectiveness of local exhaust ventilation (LEV) in controlling engineered nanomaterial emissions during reactor cleanout operations. PMID- 18432479 TI - Reduced short-term memory capacity in Alzheimer's disease: the role of phonological, lexical, and semantic processing. AB - Individuals with Alzheimer's disease (AD) are often reported to have reduced verbal short-term memory capacity, typically attributed to their attention/executive deficits. However, these individuals also tend to show progressive impairment of semantic, lexical, and phonological processing which may underlie their low short-term memory capacity. The goals of this study were to assess the contribution of each level of representation (phonological, lexical, and semantic) to immediate serial recall performance in 18 individuals with AD, and to examine how these linguistic effects on short-term memory were modulated by their reduced capacity to manipulate information in short-term memory associated with executive dysfunction. Results showed that individuals with AD had difficulty recalling items that relied on phonological representations, which led to increased lexicality effects relative to the control group. This finding suggests that patients have a greater reliance on lexical/semantic information than controls, possibly to make up for deficits in retention and processing of phonological material. This lexical/semantic effect was not found to be significantly correlated with patients' capacity to manipulate verbal material in short-term memory, indicating that language processing and executive deficits may independently contribute to reducing verbal short-term memory capacity in AD. PMID- 18432478 TI - Phonological and semantic strategies in immediate serial recall. AB - It has been suggested that certain theoretically important anomalous results in the area of verbal short-term memory could be attributable to differences in strategy. However there are relatively few studies that investigate strategy directly. We describe four experiments, each involving the immediate serial recall of word sequences under baseline control conditions, or preceded by instruction to use a phonological or semantic strategy. Two experiments varied phonological similarity at a presentation rate of one item every 1 or 2 seconds. Both the control and the phonologically instructed group showed clear effects of similarity at both presentation rates, whereas these were largely absent under semantic encoding conditions. Two further experiments manipulated word length at the same two rates. The phonologically instructed groups showed clear effects at both rates, the control group showed a clear effect at the rapid rate which diminished with the slower presentation, while the semantically instructed group showed a relatively weak effect at the rate of one item per second, and a significant reverse effect with slower presentation. The latter finding is interpreted in terms of fortuitous differences in inter-item rated associability between the two otherwise matched word pools, reinforcing our conclusion that the semantically instructed group were indeed encoding semantically. Implications for controlling strategy by instruction are discussed. PMID- 18432480 TI - The neural bases of the constructive nature of autobiographical memories studied with a self-paced fMRI design. AB - In Conway and Pleydell-Pearce's model (2000), autobiographical memories are viewed as transitory mental representations, more often generated in an effortful way. An important claim of the model concerns the dynamic process that evolves over time, from the left prefrontal areas to posterior regions, to retrieve specific memories. The present work aims at investigating, using fMRI, the temporal distribution of effortful autobiographical memory construction. In addition, a self-paced design was implemented to elucidate the question of the timing window required to evoke recollections. The results showed a large pattern of brain regions, which included the two major poles of activation predicted by Conway and Pleydell-Pearce's model. Likewise, we were able to detect the earlier implication of the left dorso-lateral prefrontal cortex, by comparison with posterior structures, which seemed to confirm its involvement in the effortful retrieval process. Finally, the self-paced procedure allowed us to refine the timing window necessary to construct past events. PMID- 18432481 TI - Identity recognition and happy and sad facial expression recall: influence of depressive symptoms. AB - Relatively few studies have examined memory bias for social stimuli in depression or dysphoria. The aim of this study was to investigate the influence of depressive symptoms on memory for facial information. A total of 234 participants completed the Beck Depression Inventory II and a task examining memory for facial identity and expression of happy and sad faces. For both facial identity and expression, the recollective experience was measured with the Remember/Know/Guess procedure (Gardiner & Richardson-Klavehn, 2000). The results show no major association between depressive symptoms and memory for identities. However, dysphoric individuals consciously recalled (Remember responses) more sad facial expressions than non-dysphoric individuals. These findings suggest that sad facial expressions led to more elaborate encoding, and thereby better recollection, in dysphoric individuals. PMID- 18432482 TI - On the costs of refocusing items in working memory: a matter of inhibition or decay? AB - The present study investigates the mental processes that are applied to previously attended items of working memory. In an object-switching task, participants counted the number of sequentially presented objects. In Experiment 1 the processing time increased when the object category switched from the prior trial compared to a repetition. More importantly, the further in the past the last instance of a current category was presented, the more processing time was necessary - an observation suggesting passive decay rather than inhibition of previously attended items. However, results differed when only two object categories were employed. Experiment 2 suggests that the lack of a clear indication of decay with small numbers of categories was due to participants' expectancy of category switches rather than repetitions. Taken together, the results suggest that working memory items become less accessible the longer they have not been attended to, when strategic processes are controlled. PMID- 18432483 TI - Why do I always have the best ideas? The role of idea quality in unconscious plagiarism. AB - Groups of individuals often work together to generate solutions to a problem. Subsequently, one member of the group can plagiarise another either by recalling that person's idea as their own (recall-own plagiarism), or by generating a novel solution that duplicates a previous idea (generate-new plagiarism). The current study examines the extent to which these forms of plagiarism are influenced by the quality of the ideas. Groups of participants initially generated ideas, prior to an elaboration phase in which idea quality was manipulated in two ways: participants received feedback on the quality of the ideas as rated by independent judges, and they generated improvements to a subset of the ideas. Unconscious plagiarism was measured in recall-own and generate-new tasks. For recall, idea improvement led to increased plagiarism, while for the generate-new task, the independent ratings influenced plagiarism. These data indicate that different source-judgement processes underlie the two forms of plagiarism, neither of which can be reduced simply to memory strength. PMID- 18432484 TI - Remembering talk: individual and gender differences in reported speech. AB - This study examined individual and group differences in the nature and frequency of reports of past speech in the autobiographical memories of young adults. A sample of 108 participants (60 females, 48 males) responded in writing to six memory prompts. They also completed the Five Factor Inventory (Costa & McCrae, 1992) and the Personal Attributes Questionnaire (Spence & Helmreich, 1978). The frequency with which participants used reported speech was correlated with agreeableness, openness, and expressivity; however, regression analyses indicated that narrators' gender alone was the best predictor. Females used more reported speech than did males. The findings suggest that recollections of past speech are an under-appreciated yet important component of autobiographical memory. PMID- 18432485 TI - Psychotropic placebos reduce the misinformation effect by increasing monitoring at test. AB - A psychotropic placebo can help people resist the misinformation effect, an effect thought to be caused by a shift to more stringent source monitoring. When this shift occurs has been unclear. To address this issue we gave some people - but not others - a phoney cognitive-enhancing drug we called R273. Shortly afterwards, everyone took part in a misinformation effect experiment. To gather evidence about source monitoring we surreptitiously recorded time to read the misleading postevent narrative, and response time at test. Our findings suggest that people shifted to more stringent source monitoring at test. Moreover, people with higher working memory capacity (WMC) performed better than people with lower WMC - but only when they were told they had received R273, a finding that fits with research showing that WMC can confer advantages in situations demanding effortful control, but not when automatic heuristics suffice. PMID- 18432486 TI - The cultural life script and life story chapters contribute to the reminiscence bump. AB - A total of 59 older Danes recorded five life story memories. They divided their life story into chapters and provided their age for the start and end of each chapter. Life story memories were coded for whether they were placed at the start or end of chapters and for their correspondence to the cultural life script. Chapters and life story memories showed a bump in terms of an increased recall of life story memories and chapters between ages 6 and 30. Chapter start and end memories, more frequently than other memories, referred to prominent cultural life script events. The bump was significantly stronger for memories that referred to both prominent cultural life script events and chapter starts or ends. The findings suggest that the cultural life script helps to identify beginning and ends of chapters in autobiographical memory, and that both the cultural life script and organisation in terms of chapters influence the recall of life story memories and may help to explain the bump. PMID- 18432487 TI - The co-witness misinformation effect: memory blends or memory compliance? AB - For 30 years hundreds of researchers have shown participants videos and slide sequences of events, presented the participants with misleading information, and found that this misinformation distorted their memories. The purpose of this study was to establish whether those misled participants are reporting a memory blend of the two sources of information or whether they are simply complying with the post-event information. A total of 92 participants were shown one of two versions of six different videos, which included some subtle differences. After having watched each video individually, participants were paired with someone who had seen the other version and they discussed the clips together. They then individually answered questions about the videos, and their responses showed that some of the distorted memories were blends of the original information and the post-event information. The implications of these findings are discussed. PMID- 18432488 TI - False recognition following study of semantically related lists presented in jumbled word form. AB - Three experiments explored a jumbled word effect in false recognition. Lists of theme-related items were presented in word or nonword form. Results indicated that critical lures semantically related to studied items were falsely recognised regardless of whether they were presented as words or nonwords. High false recognition rates to either SLEEP or SELEP following study of an appropriate theme list of items in nonword form should only occur if nonwords are recoded at study. With study conditions conducive to recoding, jumbled words induced false recognitions based on semantic associations among their respective base words. Disrupting a recoding process by creating "difficult" letter rearrangements for jumbled words (Experiment 2) appeared to eliminate the false recognition effect. In Experiment 3, presentation durations ranged from 110 ms to 880 ms. Although there was little evidence of a semantic false recognition effect at the fastest presentation rate, the brief durations appeared to be effective in eliminating the effect when items were studied in nonword form. These results appear to be consistent with an encoding activation/retrieval monitoring model. PMID- 18432489 TI - Epidemiological economics. PMID- 18432490 TI - An introduction to decision analysis in the economic evaluation of the prevention and treatment of vision-related diseases. AB - Economic evaluation in the form of reports of cost-effectiveness of the treatment and prevention of disease has only recently found widespread application in the visual sciences. While economic evaluation takes a number of forms: cost minimization analysis, cost-benefit analysis, and cost-effectiveness analysis--it is the latter that is seen most often in the evaluation of vision-related health programs. Cost-effectiveness analysis is in particular seen most commonly in its very particular form of cost-utility analysis. Decision analysis is the analytic method most commonly used to perform cost-effectiveness analysis. In decision analysis, the expected cost and effectiveness of a health program are estimated in a rigorous fashion. In this report, we take the reader through the process of decision analysis including building the tree; populating the model with parameters for risk, cost and benefit; estimating expected cost and benefit; and deterministic and probabilistic sensitivity analysis. Examples employed include prominent studies of the cost-effectiveness of photodynamic therapy for treatment of neovascular macular degeneration and the treatment ocular hypertension to prevent glaucoma. PMID- 18432492 TI - The economic costs and benefits of dog guides for the blind. AB - PURPOSE: To document the economic costs and benefits associated with providing dog guide services for blind individuals able to benefit from them. METHODS: This study estimates the annual cost of dog guide services accounting for cost offsets associated with reduced informal and formal care costs over the working life of the animal (8 years). We estimated the cost per dog guide trained using previously unpublished survey data from dog guide training schools in the United States. We also estimated the incremental economic benefits as the reduction in costs associated with formal and informal care using published studies and a set of reasonable assumptions. Costs were discounted to 2006$ using a 3% discount rate. RESULTS: We found the average total cost per dog guide over its working life was $40,598, of which $21,568 were off-set by reductions in other costs. The costs associated with dog guides included $35,536 in dog acquisition costs and $5,061 for annual maintenance over the animal's working life. The economic benefits included $16,324 and $5,244 in reduced formal and informal care costs, respectively. The average net cost of dog guide ownership per year over the working life of the animal was $2,379. CONCLUSIONS: Using available information and reasonable assumptions, this study documents the costs of dog guides accounting for a limited number of cost off-setting elements. However, given limited available evidence, further study of the impact of guide dogs on the lives of blind individuals who use them should be conducted to validate this study's results. PMID- 18432491 TI - Economic evaluation of blind rehabilitation for veterans with macular diseases in the Department of Veterans Affairs. AB - PURPOSE: The Department of Veterans Affairs (VA) Low Vision Intervention Trial (LOVIT) developed an outpatient low-vision programme for patients with macular diseases providing low-vision rehabilitation comparable to VA inpatient blind rehabilitation centres (BRCs). This programme targets veterans who do not need or chose not to participate in a comprehensive inpatient blind rehabilitation programme. We examined costs and consequences using veterans in LOVIT and comparable veterans in an inpatient BRC. METHODS: We compared costs and consequences between treatment patients who participated in LOVIT, a two-site randomized clinical trial, and a sample of comparable patients who received treatment at a VA inpatient BRC. We measured consequences as the change in functional visual ability from baseline to follow-up (LOVIT: 4 months after randomization; BRC: 3 months after discharge) using the VA Low Vision Visual Functioning Questionnaire (VA LV VFQ-48). RESULTS: There were 55 LOVIT and 121 BRC patients for our analyses. Average costs were $38,627.3 higher for BRC patients ($5,054.4 +/- $404.7 SD for LOVIT vs. $43,681.7 +/- $8,853.6 SD for BRC, p < 0.0001). Thus, the BRC cost $38,627.3 per patient more than the LOVIT programme (95% CI: $17,414 to $273,482). There was a greater improvement in overall visual ability, mobility, and visual motor skill scores for BRC patients; however, there was no significant difference in improvement in reading ability or visual information processing scores. CONCLUSIONS: As VA increases outpatient blind rehabilitation services, LOVIT provides a model for expanding outpatient low-vision rehabilitation services for veterans at substantially lower costs than current inpatient BRC services. PMID- 18432493 TI - Reversal in gender valuations of cataract surgery after the implementation of free screening and low-priced high-quality surgery in a rural population of southern China. AB - PURPOSE: To assess the impact of community outreach and the availability of low cost surgeries [500 Renminbi (RMB) or 65 United States dollars (US$) per surgery] on the willingness to pay for cataract surgery among male and female rural dwelling Chinese. METHODS: Cross-sectional willingness-to-pay surveys were conducted at the initiation of a cataract outreach programme in June 2001 and then again in July 2006. Respondents underwent visual acuity testing and provided socio-demographic data. RESULTS: In 2001 and 2006, 325 and 303 subjects, respectively, were interviewed. On average the 2006 sample subjects were of similar age, more likely to be female (p < 0.01), illiterate (p < 0.01), and less likely to come from a household with annual income of less than US$789 (62% vs. 87%, p < 0.01). Familiarity with cataract surgery increased from 21.2% to 44.4% over the 5 years for male subjects (p < 0.01) and 15.8%-44.4% among females (p < 0.01). The proportion of respondents willing to pay at least 500 RMB for surgery increased from 67% to 88% (p < 0.01) among male subjects and from 50% to 91% (p < 0.01) among females. CONCLUSIONS: Five years of access to free cataract testing and low-cost surgery programmes appears to have improved the familiarity with cataract surgery and increased the willingness to pay at least 500 RMB (US$65) for it in this rural population. Elderly women are now as likely as men to be willing to pay at least 500 RMB, reversing gender differences present 5 years ago. PMID- 18432494 TI - Assessing participation in daily living and the effectiveness of rehabiliation in age related macular degeneration patients using the impact of vision impairment scale. AB - PURPOSE: To assess if the Impact of Vision Impairment (IVI) is a valid instrument to measure participation in daily activities and rehabilitation in patients with age-related macular degeneration (AMD) and varying levels of visual impairment. METHODS: Participants, recruited from a public eye hospital and low vision centers, completed the IVI questionnaire. The IVI and its three subscales were assessed for fit to the Rasch model. Unidimensionality, item fit, response category performance, and targeting of items to patients were assessed. Confirmatory factor analysis (CFA) was used to assess the three-factor model of the IVI in this sample of AMD patients. RESULTS: 219 patients (mean +/- SD age = 83.5 +/- 7.4 yr) were recruited. Of these, 22%, 55% and 23% had mild (< 6/12 6/18), moderate (< 6/18-6/60) and severe (< 6/60) vision loss, respectively. The IVI total and three subscales displayed discrete thresholds indicating that the respondents understood the response categories. The IVI items fitted the scale and unidimensionality was established. Person separation reliability for the IVI score was substantial (0.94) indicating that the scale can discriminate between several groups of AMD patients. The IVI items were significantly targeted to the AMD patients with the means of the two distributions shown to be very close (0.0 and 0.1, respectively). Substantial targeting was also evident for the subscales. Poorer visual acuity was significantly associated (ANOVA; F (2, 216) = 23.4; p < 0.001) with greater restriction of participation suggesting that the IVI has substantial construct validity. CFA supported the IVI three-factor model which includes items from the "emotional well-being, "reading and accessing information" and "mobility and independence" subscales. CONCLUSIONS: Clinicians and researchers can reliably use the IVI to assess the impact on daily life and the effectiveness of clinical trials and rehabilitation interventions in patients with AMD across a range of vision loss. PMID- 18432495 TI - Defining the content for a new quality of life questionnaire for students with low vision (the Impact of Vision Impairment on Children: IVI_C). AB - PURPOSE: To obtain statements describing the impact of vision impairment on participation in every day activities of school-aged children (8-18 years) capturing their opinion, as the primary step in developing a pediatric vision related quality of life instrument. METHODS: Separate focus groups for students with low vision, parents and teachers plus in-depth individual interviews were conducted in Victoria, Australia. Focus groups followed a topic guide, were digitally audio recorded and transcribed verbatim. Statements were identified, coded and ascribed into major themes. RESULTS: 102 participants provided just over 2600 statements (an average of 146 statements per verbatim transcript) which were classified into 5 themes: school/specialist instruction, social interaction, family, community and vision impairment peer interaction. The identified areas likely to facilitate good participation in every day activities, and accounted for 79% of the statements were class teacher knowledge and specialist support (40%) and social interaction (39%). The student perspective focused on communication skills and orientation and mobility whereas the specialist instructors' emphasis was on specialist support. Emphases across the themes varied between focus groups and highlight the benefit of including the perspectives of all stakeholders in questionnaire design. CONCLUSIONS: The multi perspective development strategy for a new questionnaire has established that it is crucial to integrate different stakeholders' perspectives. By doing so, it will be possible to develop a balanced questionnaire. PMID- 18432496 TI - Survey of older drivers' experiences with Florida's mandatory vision re-screening law for licensure. AB - PURPOSE: To document the license renewal experiences of Florida drivers ages 80 and older who must pass a visual acuity test when renewing their driver's license. METHODS: Study participants (n = 1,242 drivers) were contacted via telephone and completed a survey regarding their experiences with the license renewal process. RESULTS: The majority (80.2%) of those eligible for license renewal reportedly attempted to do so and 88.0% succeeded the first time they tried. A large percentage of drivers (88%) who failed the vision test said they sought treatment, and 77.6% of drivers who reattempted renewal reportedly passed the test. About half of drivers who did not seek renewal said they thought they would fail the vision test. The majority of those choosing not to renew their license (99.5%) reported using transportation alternatives. CONCLUSIONS: This study suggests that the Florida vision screening re-licensure law is not a deterrent to seeking license renewal for the > or = 80-year-old population. Furthermore, only a small percentage of Florida drivers ages > or = 80 years and older reported that they failed the visual acuity screening test and were denied license renewal. PMID- 18432497 TI - Visual acuity and mortality in older people and factors on the pathway. AB - PURPOSE: To examine vision as a predictor of mortality in older people and the role of mobility, depressed mood, chronic diseases, body mass index, physical activity and injurious accidents in this possible association. METHODS: 223 persons aged 75 and 193 persons aged 80 years at the baseline participated in visual acuity measurements. Visual acuity (VA) of < 0.3 in the better eye was defined as visual impairment, VA of > or = 0.3 but < or = 0.5 as lowered vision and VA > 0.5 as normal VA. Death dates were received from the official register. Cox regression models were used to determine the relative risks of mortality and to study what factors lie on the pathway from poor vision to mortality. RESULTS: Over the 10-year follow-up, 107 (48%) persons aged 75 years and 138 (72%) aged 80 years at the baseline died. The risk for mortality among the 75-year-olds with lowered vision was 1.98 (95 % CI 1.25-3.13) and with visual impairment 1.90 (95% CI 1.12-3.20) compared to those with normal VA. Lower walking speed, physical inactivity, cardiovascular diseases, injurious accidents, diabetes and depressed mood each attenuated the risk markedly. Nevertheless, lowered vision remained a significant predictor of mortality even after including all these variables in the model. Among the 80-year-olds vision did not correlate with mortality. CONCLUSIONS: Lowered vision and severe visual impairment predicted mortality in the 75-year-old but not 80-year-old population. The increased risk was partially explained by lower walking speed, physical inactivity, cardiovascular diseases, depressed mood, diabetes and injurious accidents. PMID- 18432498 TI - Faggot formation in mature neutrophils and metamyelocytes in acute myeloid leukemia without maturation. AB - The authors report a rare case of acute myeloid leukemia (AML) M1 with faggot formation in mature neutrophils and metamyelocytes. Although Auer rods in mature neutrophils are occasionally experienced, they are usually found in AML M2, M3, or M4 cases, but not in M1 cases. In addition, faggot formation in mature neutrophils, as seen in this case, is considered to be particularly unusual because most previously reported cases tended to show simple Auer rods except for AML M3 cases. This case also showed trisomy 4 in its karyotype, although its relationship with faggot formation remains unclear. PMID- 18432499 TI - Fatal sibling cases of familial hemophagocytic lymphohistiocytosis (FHL) with MUNC13-4 mutations: case reports. AB - The authors report here sibling cases of familial hemophagocytic lymphohistiocytosis (FHL) type 3 that took fatal courses despite intensive treatment. The older brother achieved remission by immunochemotherapy, but a central nervous system lesion occurred before the introduction of allogeneic hematopoietic stem cell transplantation (allo-HSCT). The patient died on day +1 of allo-HSCT due to progression of the disease. The younger brother developed symptoms of hemophagocytic lymphohistiocytosis mimicking neonatal hemochromatosis at birth. He died without a chance to receive allo-HSCT. Both siblings showed low natural killer cell (NK) activity and the compound heterozygous Munc13-4 gene mutations 1596+1 and 1723insA were identified postmortem in the younger brother. With recent progress in the molecular diagnosis of FHL, prompt and most appropriate therapeutic measures should be introduced to improve the prognosis of FHL patients. PMID- 18432500 TI - Spontaneous remission of acute lymphoblastic leukemia with mediastinal mass. AB - Spontaneous remission/regression of cancer is defined as partial or complete disappearance of malignant disease temporarily or permanently in the absence of medical treatment. This event is named as spontaneous regression for solid tumors and spontaneous remission for leukemia. The authors report the case of a girl aged 4 years and 3 months, who presented with mediastinal mass and leukemic findings in the bone marrow both of which reappeared after spontaneous regression and remission, respectively. PMID- 18432501 TI - Endocrinopathies in patients with transfusion-dependent beta-thalassemia. AB - Thalassemia is common in Iran. Appropriate therapy for this disease includes a regular blood transfusion and chelation therapy. However, in this approach patients will inevitably experience side effects, particularly iron overloads in critical organs, including heart, ductless glands, and liver. This study attempted to determine prevalence of adenoidal abnormality between Iranian thalassemia patients for prediagnosis and to offer necessary medical measures. This is a descriptive nonrandomized study and included all the patients suffering from thalassemia major referring to medical centers linked with the Iranian blood transfusion organization from January 2004 to January 2005. All patients were sampled for CBC, FBS, 2-h BS, HbAlC, liver function, renal function, and endocrine disease. Initially, reports of adenoidal experiments as well as other associated parameters were provided from medical records. A total of 437 patients enrolled in the study: 5.4% suffered from diabetes, 1% had hypothyroid, and 1 person showed hypoparathyroidism. The mean levels of ferritin in diabetic and nondiabetic groups and hypothyroid and nonhypothyroid groups were not significantly different. The mean levels of ferritin among various age groups also were not significantly different. Results of a comparison between present research and similar studies conducted throughout Iran and those performed abroad on adenoidal abnormalities in thalassemia patients show that subject patients of this study statistically suffered from fewer difficulties than diabetes patients in other studies. PMID- 18432502 TI - Assessment of febrile neutropenia episodes in children with acute leukemia treated with BFM protocols. AB - The authors overviewed 239 febrile neutropenia (FN) episodes in 82 pediatric leukemia cases treated with BFM treatment protocols. FN was observed mostly during consolidation therapy. Mucositis was the most identified focus; gram negative microorganisms were the most identified pathogens. Five patients developed invasive fungal infections. Fever resolved after mean 5.3 days and mean antibiotic administration time was 12.7 days. Addition of G-CSF to antimicrobial therapy shortened the duration of neutropenia, but it did not affect duration of fever resolution and antibiotic administration. The duration of neutropenia, fever resolution, and antibiotic administration was significantly longer in children with acute myeloid leukemia. The authors conclude that children with acute leukemia have severe prolonged neutropenia and are in high risk. In these patients, prediction of the risk of bacteremia based on clinical and laboratory features is important for immediate empiric broad-spectrum antimicrobial therapy and for higher survival rate. PMID- 18432503 TI - Hemarthrosis due to a rare cause of hemorrhagic diathesis: Ehlers-Danlos syndrome. AB - The authors report a case of hemarthrosis complicated by severe anemia related to a congenital connective tissue disease: Ehlers-Danlos syndrome. A boy fell down and suffered tumefaction of both knees with bilateral rupture of the rotula tendon. He underwent surgical reinsertion of each tendon on the rotula. He later showed an unexpected ongoing hematic effusion, with severe anemia. He was screened for coagulation disorders with no results. On taking a more detailed history and investigating the patient's phenotypical features, the authors diagnosed Ehlers-Danlos syndrome, hypermobile variant. The hemarthrosis and anemia were thus concluded to be consequences of excessive tissue fragility due to a congenital connective tissue disease. PMID- 18432504 TI - Chemotherapy-induced acral erythema in a pediatric patient with acute monoblastic leukemia. AB - Chemotherapy-induced acral erythema or palmoplantar erythrodysesthesia syndrome is a well-defined reaction to some of the chemotherapeutic agents such as methotrexate, cytarabine, doxorubicin, fluorouracil, and bleomycin. This reaction is characterized by symmetrical, well-demarcated, painful erythema of the palms and soles, which may progress to desquamation. The authors present a case of acral erythema in a young patient with acute monoblastic leukemia to emphasize this high-dose chemotherapy-induced side effect, which is rarely seen in children and is usually self-limited. PMID- 18432505 TI - Granulomatous reaction in mediastinal B-cell non-Hodgkin lymphoma and intracardiac thrombosis. AB - Epithelioid cell granulomas may be associated with several neoplasms. Lymphomas may mimic or are associated with epithelioid granulomas. In this article the authors report a child with granulomatous reaction in B cell non-Hodgkin lymphoma and intracardiac thrombosis. Although cancer alone is a risk factor for thromboembolism, thrombosis is a multifactorial disorder with both hereditary and acquired risk factors. This is the first reported case of intracardiac thrombosis with MTHFR A1298C and factor XIII V34L mutations together with granulomatous reaction in non-Hodgkin lymphoma. PMID- 18432506 TI - Double heterozygosity for Hb New York [beta 113 GTG-->GAG; VAL-->GLU] and beta degrees-thalassemia mutations manifests as a thalassemia trait. AB - An extended family with three individuals affected by two different forms of double heterozygosity for beta-thalassemia and Hb New York is reported. Double heterozygosity of Hb New York [beta 113 GTG-->GAG; VAL-->GLU] and beta degrees codon 17 was detected in a fetus following prenatal screening for thalassemia. The father and a paternal aunt were also found to be heterozygous for Hb New York and beta degrees IVSII-654. Both adults had clinical and hematological features consistent with beta-thalassemia trait. The affected child was followed up after birth and manifested the typical course of a thalassemia trait, with no signs of organomegaly or overt hemolysis. Observations strongly suggest that double heterozygosity of Hb New York and beta degrees thalassemia has mild, if any, clinical symptoms, and is not an indication of therapeutic abortion when detected antenatally. PMID- 18432507 TI - Antiphospholipids antibodies in a 12-month-old presenting with idiopathic thrombocytopenic purpura. AB - A 12-month-old Caucasian female with a history of recurrent ear infections presented to the emergency room with petechiae, severe thrombocytopenia (4000/microL), and abnormally prolonged activated partial thromboplastin time. Further autoantibody investigation detected antinuclear antibodies, anti-double stranded DNA, and antiphospholipid antibodies. Platelet count, in response to intravenous immunoglobulin infusion, increased to more than 100 x 10(3) plt/microL. At 6-month follow-up, no positive autoantibodies were detected. PMID- 18432508 TI - Fatal visceral varicella-zoster virus infection without skin involvement in a child with acute lymphoblastic leukemia. AB - A 5-year-old girl with acute lymphoblastic leukemia in remission suffered from fatal visceral varicella-zoster virus (VZV) infection after the oral administration of a high-dose dexamethasone. She abruptly developed fulminant hepatitis and disseminated intravascular coagulation, and died 3 days later. VZV DNA and antigens were detected in the peripheral blood (6 x 10(8) copies/mL) and a postmortem liver specimen, respectively. The exposure to VZV was not confirmed and no skin lesions were observed. VZV infection should be considered in patients with unexplained liver dysfunction under severe immunosuppressive condition, even in the absence of viral exposure and skin involvement. PMID- 18432509 TI - Re: Recurrent immune thrombocytopenic purpura in children. PMID- 18432510 TI - Motor function in adults with Asperger's disorder: a comparative study. AB - In the original description of Asperger's disorder (AD), clumsiness was an associated feature. Several studies of children have shown deficits in motor control, whereas research regarding adults is scarce. The aim of the present study was to compare motor function in adults with AD, with a normal comparison group. Gross and fine motor skills were examined by a standardized, norm referenced test developed for children, but also used in young adults, the Bruininks-Oseretsky Test of Motor Proficiency (BOTMP). In addition, a questionnaire regarding the participants' physical activity during leisure time was administered. We found that adults (age 21-35) with AD (N = 15) performed significantly worse than the normal comparison group (N = 29) in six of eight subtests in the BOTMP. Males with AD were less physically active than males in the comparison group. Among females, physical activity did not differ between the groups. There was a positive association between physical activity and gross motor function in the AD group. Participants with AD were encouraged by the assessments. Physical coaching may be an important future field for improving quality of life in adults with AD. PMID- 18432511 TI - Breathing retraining in patients with chronic fatigue syndrome: a pilot study. AB - The study aimed to 1) examine the point prevalence of asynchronous breathing in chronic fatigue syndrome (CFS) patients; 2) examine whether CFS patients with an asynchronous breathing pattern present with diminished lung function in comparison with CFS patients with a synchronous breathing pattern; and 3) examine whether one session of breathing retraining in CFS patients with an asynchronous breathing pattern is able to improve lung function. Twenty patients fulfilling the diagnostic criteria for CFS were recruited for participation in a pilot controlled clinical trial with repeated measures. Patients presenting with an asynchronous breathing pattern were given 20-30 minutes of breathing retraining. Patients presenting with a synchronous breathing pattern entered the control group and received no intervention. Of the 20 enrolled patients with CFS, 15 presented with a synchronous breathing pattern and the remaining 5 patients (25%) with an asynchronous breathing pattern. Baseline comparison revealed no group differences in demographic features, symptom severity, respiratory muscle strength, or pulmonary function testing data (spirometry). In comparison to no treatment, the session of breathing retraining resulted in an acute (immediately postintervention) decrease in respiratory rate (p < 0.001) and an increase in tidal volume (p < 0.001). No other respiratory variables responded to the session of breathing retraining. In conclusion, the present study provides preliminary evidence supportive of an asynchronous breathing pattern in a subgroup of CFS patients, and breathing retraining might be useful for improving tidal volume and respiratory rate in CFS patients presenting with an asynchronous breathing motion. PMID- 18432512 TI - Effect of penetrating trunk trauma on the recovery of adult survivors: a pilot study. AB - Gunshot and/or stab wounds to the trunk are injuries seen in South African hospitals. Patients are managed in the intensive care unit. Prolonged mechanical ventilation with immobilization results in some degree of muscle dysfunction. Our goal was to determine if patients recover adequately spontaneously following critical illness. No formal rehabilitation programmes exist in South Africa for these patients following discharge. A prospective, observational study was conducted. Patients were recruited from three ICUs in Johannesburg. Lung function tests, dynamometry, quality of life, 6-minute-walk, and oxygen uptake tests were performed over 6 months following discharge from the hospital. The control group consisted of existing data for healthy volunteers. Distance walked during 6 minute-walk test was significantly reduced for the study group compared to the control group (1 month [p = 0.00251]; 6 months [p = 0.0355]). At 1 month there was a significant reduction in quadriceps and triceps strength for the study group compared to the control group (p = 0.0089; p = 0.0246, respectively). Quadriceps strength remained significantly reduced for the study group (3 months [p = 0.0489]). No difference in muscle strength was detected between the groups at 6 months. Actual and predicted residual volumes differed significantly for the study group (1 month [p = 0.0034]; 6 months [p = 0.0157]). A trend of muscle weakness (to 3 months), poor exercise capacity, and abnormal lung volumes was identified. A formal rehabilitation programme may be needed to address these disabilities. PMID- 18432513 TI - Balance-related efficacy compared with balance function in patients with acute stroke. AB - It is well known that patients who have suffered a stroke have problems with balance and have feelings of unsteadiness. The aim of this study was to analyse the correlation between how patients estimate their perceived confidence in task performance without falling and the objective assessment of balance made by a physiotherapist, and whether the Falls Efficacy Scale (FES(S)) is to be recommended for use in patients in the acute phase of stroke. Sixty patients divided into two groups assessed their belief to perform daily life activities without fear of falling using the FES(S) and were assessed by using the Berg Balance Scale (BBS) and Timed Up and Go (TUG) by a physiotherapist. Group 1 assessed themselves before, whereas Group 2 assessed themselves after the objective assessment. The correlation between the FES(S) and the TUG was moderate to good, but these two scales consider different aspects and dimensions of balance, ability, and belief and are therefore not interchangeable. The correlation between FES(S) and BBS was low to moderate. The conclusion of this study was that the FES(S) is not to be recommended as a single measurement in the acute phase of stroke because it does not measure actual balance function. PMID- 18432514 TI - Determining the minimal number of cyclic passive stretch repetitions recommended for an acute increase in an indirect measure of hamstring length. AB - The purpose of this study was to determine the minimal number of cyclic passive stretch repetitions required to induce an acute increase in hamstring length via an indirect measure of hamstring length. Eighteen healthy males (aged 19-37 years) were placed in a standardized position. Knee flexion range of motion (ROM) measurements were recorded following ten 15-second passive stretch repetitions. Analysis of variance measures (p < 0.05) and post hoc analysis showed that significant increases in knee extension ROM diminished at the fifth stretch repetition. The greatest single increase in knee extension ROM occurred after the first stretch. The results of this study indicate that passive stretching beyond five repetitions results in insignificant gains in hamstring length (measured via knee extension ROM) and are probably unnecessary. PMID- 18432515 TI - A clinical assessment tool used for physiotherapy students--is it reliable? AB - Educational institutions providing professional programs such as physiotherapy must provide high-quality student assessment procedures. To ensure that assessment is consistent, assessment tools should have an acceptable level of reliability. There is a paucity of research evaluating the reliability of clinical assessment tools used for physiotherapy students. This study evaluated the inter- and intrarater reliability of an assessment tool used for physiotherapy students during a clinical placement. Five clinical educators and one academic participated in the study. Each rater independently marked 22 student written assessments that had been completed by students after viewing a videotaped patient physiotherapy assessment. The raters repeated the marking process 7 weeks later, with the assessments provided in a randomised order. The interrater reliability (Intraclass Correlation Coefficient) for the total scores was 0.32, representing a poor level of reliability. A high level of intrarater reliability (percentage agreement) was found for the clinical educators, with a difference in section scores of one mark or less on 93.4% of occasions. Further research should be undertaken to reevaluate the reliability of this clinical assessment tool following training. The reliability of clinical assessment tools used in other areas of physiotherapy education should be formally measured rather than assumed. PMID- 18432516 TI - Reliability of inclinometer and goniometric measurements of hip extension flexibility using the modified Thomas test. AB - Our objectives were to examine interrater reliability for inclinometer and goniometric measurements of hip extension flexibility and the intra- and interrater reliability between instruments when measuring hip extension flexibility with the modified Thomas test. The modified Thomas test is frequently used by clinicians to assess hip extension flexibility. Studies examining the reliability of measures of the modified Thomas test are limited. This study included 42 healthy subjects. The modified Thomas test was performed on each subject to quantify hip extension flexibility by using an inclinometer and a goniometer. High interrater reliability was determined for each instrument (r = 0.91-0.93; ICC = 0.89-0.92). High intrarater parallel-forms reliability was found for each examiner between the instruments (r = 0.089-0.92; ICC = 0.91-0.93). High interrater parallel-forms reliability also was found between instruments (r = 0.86-0.93; ICC = 0.86-0.92). The high correlations obtained in this study suggest that the inclinometer and the goniometer are reliable instruments for measuring hip extension flexibility. The results also indicate that the two instruments can be used interchangeably for measuring hip extension flexibility. PMID- 18432517 TI - Effects of heat on camel platelet structure and function-a comparative study with humans. AB - Camels and many other desert animals are uniquely adapted to conserve water and other fluids in order to survive intense heat for long periods. Earlier studies have suggested that human platelets may be the trigger for the coagulopathy involved in heat prostration and stroke. The present study has compared the resistance of camel and human platelets to heat in order to see if they might help to protect camels from the effects of high body temperature for prolonged periods. The findings demonstrate that camel platelets are significantly less sensitive to heat than human platelets. Temperatures (43 degrees C-45 degrees C) that cause human cells to undergo marked structural alterations and lose their ability to spread and aggregate have no effect on camel platelets. Even higher temperatures (50 degrees C) that destroy human platelets have minor effects on camel cells and do not seriously compromise their function. Temperatures of 55 degrees C do destroy camel platelets and their functional capability. The resistance of camel platelets to heat may help protect camels from the effects of extreme body temperature and dehydration, which are everyday conditions in the desert. PMID- 18432518 TI - External calcium facilitates signalling, contractile and secretory mechanisms induced after activation of platelets by collagen. AB - Platelet activation leads to the initiation of intracellular signalling processes, many of which are triggered by Ca2+. We have studied the involvement of exogenous Ca2+ in platelet response to collagen activation. Platelet suspensions were prepared with and without adding external calcium in the suspension buffers. Activation with collagen (Col-I) was carried out, before and after incubation with cytochalasin B (Cyt-B) to block the actin assembly and the cytoskeletal reorganization. We evaluated changes in (i) tyrosine phosphorylation of proteins, in platelet lysates and associated with the cytoskeletal fraction, (ii) the association of contractile proteins to the cytoskeleton, (iii) expression of intraplatelet substances at the surface, and (iv) cytosolic Ca2+ levels ([Ca2+]i). Ultrastructural evaluation of platelets by electron microscopy was also performed. Platelet activation by Col-I in the absence of added Ca2+ was followed by mild association of actin and other contractile proteins, low phosphorylation of proteins at tyrosine residues, lack of expression of intraplatelet substances at the membrane, and absence of aggregation. In the presence of millimolar Ca2+, Col-I induced intense actin filament formation with association of contractile proteins with the cytoskeleton, resulting in profound morphological changes. Under these conditions, Col-I induced signalling through tyrosine phosphorylation, with increases in the [Ca2+]i, release of intragranule content and aggregation. Inhibiting actin polymerization with Cyt-B prevented all these events. Our data indicates that platelet activation by collagen requires external Ca2+. Studies with Cyt-B indicate that assembly of new actin and cytoskeleton-mediated contraction, both dependent on exogenous Ca2+, are key events for platelet activation by collagen. In addition, our results confirm that entrance of exogenous Ca2+ depends on a functional cytoskeleton. PMID- 18432519 TI - The interaction of factor VIIa with rehydrated, lyophilized platelets. AB - The experiments presented here were undertaken to determine if factor VIIa (rFVIIa, the Novo Nordisk product NovoSeven) will directly bind to rehydrated, lyophilized (RL) platelets for the formation of a catalytic surface with an enhanced ability to generate thrombin. The interaction between rFVIIa and the RL platelet surface was examined by measuring equilibrium and non-equilibrium binding of the coagulation factor to the cells and by following the effects of the surface modification on the kinetics of thrombin generation. The association of rFVIIa with RL platelets was rapid with saturation occurring within minutes. Disassociation was slow, with over half of the coagulation factor remaining bound after two hours. Densities of over one million molecules of rFVIIa per RL platelet were obtained when high concentrations of rFVIIa were incubated with RL platelets. Thrombin generation measurements showed that RL platelet-bound rFVIIa was catalytically active. Thus we can expect that RL platelets, which have been shown to effectively bind to sites of vascular injury, will localize rFVIIa to wounds for an increase in therapeutic index. These studies indicate that rFVIIa RL platelets are worthy of preclinical and clinical development as an infusion agent for severe bleeding. PMID- 18432520 TI - Significance of microparticles in progressive systemic sclerosis with interstitial pneumonia. AB - We measured and compared the levels of microparticles, chemokines, cell adhesion molecules and platelet activation markers with a view to developing a better understanding of their potential contributions to the pathophysiology of progressive systemic sclerosis (PSS, scleroderma). The concentrations of all the factors in PSS patients were significantly higher than those in normal subjects. PSS patients were divided to two groups by whether they have interstitial pneumonia (IP) or not. There were no differences in the levels of soluble(s) VCAM 1, sICAM-1, sE-selectin and IL-8 between the two groups. However, there were significant between-group differences in the levels of sP-selectin, sCD40L, ENA 78, RANTES (regulated on activation normally T-cell expressed and secreted), platelet-derived microparticles (PDMPs), monocyte-derived microparticle (MDMPs) and KL-6. The level of tissue factor expression on monocytes by A23187 stimulation in PSS patients was found to be similar to that in healthy controls. Although PDMP did not induce the expression of tissue factor on monocytic cell line (THP-1) directly, the recombinant sCD40 ligand-induced expression of tissue factor on THP-1 and generation of MDMP from this cell line were enhanced by the addition of PDMPs. Our findings suggested that elevated levels of PDMPs and MDMPs may be interpreted as a sign of vascular complications in PSS patients, particularly those complicated with IP, offering a new treatment strategy in these patients. PMID- 18432521 TI - Critical roles for the actin cytoskeleton and cdc42 in regulating platelet integrin alpha2beta1. AB - The modified two-site model for platelet activation by collagen requires tight binding of platelets to collagen through integrin alpha2beta1, after its prior activation by inside-out signals initiated by GP VI. The inside-out signalling to alpha2beta1 is not well characterized although it is currently accepted that GPVI initiates signals that lead to regulation of this integrin. The aim of the study was to determine the role played by actin polymerization and the Rho family GTPase cdc42 in the regulation of alpha2beta1 integrin. We first show that GPVI- and non-GPVI-dependent signals differentially regulate distribution of alpha2beta1 receptors, where binding of platelets to collagen leads to redistribution of the integrin to areas of contact between platelet and collagen fibre. Binding of platelets to collagen also leads to activation of alpha2beta1 integrin, which is dependent upon actin polymerization and cdc42 activity, since activation is blocked by cytochalasin D and secramine A respectively. Adhesion of platelets to collagen is markedly diminished in the presence of these inhibitors, whereas adhesion to CRP- or fibrinogen-coated surfaces is not affected. Platelet aggregation to collagen, but not CRP or thrombin, is also markedly dependent upon actin polymerization and cdc42 activity. In conclusion these data suggest that actin polymerization and cdc42 are required for activation of integrin alpha2beta1, but not alpha(IIb)beta3, thereby critically regulating platelet adhesion to and activation by collagen. We therefore suggest a further modification to the current two-site two-step model for activation of platelets by collagen, where actin polymerization and cdc42 mediate a critical step in modulating alpha2beta1 activation, possibly through a positive feedback pathway from alpha2beta1 itself. PMID- 18432522 TI - Phospholipase D in human platelets: presence of isoenzymes and participation of autocrine stimulation during thrombin activation. AB - Phospholipase D (PLD), which hydrolyzes phosphatidylcholine to phosphatidic acid (PA) and choline, is present in human platelets. Thrombin and other agonists have been shown to activate PLD but the precise mechanisms of activation and PLDs role in platelet activation remains unclear. We measured thrombin-stimulated PLD activity in platelets as formation of phosphatidylethanol. Since no specific PLD inhibitors exist, we investigated possible roles for PLD in platelets by correlating PLD activity with platelet responses such as thrombin-mediated secretion and F-actin formation (part of platelet shape change). Extracellular Ca2+ potentiated thrombin-stimulated PLD, but did not stimulate PLD in the absence of thrombin. Thrombin-induced PLD activity was enhanced by secreted ADP and binding of fibrinogen to its receptors. In contrast to others, we also found a basal PLD activity. Comparison of time courses and dose responses of platelets with PLD showed many points of correlation between PLD activation and lysosomal secretion and F-actin formation. The finding of different PLD activities suggested that different PLD isoenzymes exist in platelets as reported for other cells. Here we present evidence for the presence of both PLD1 and PLD2 in platelets by use of specific antibodies with immunoblotting and immunohistochemistry. Both isoforms were randomly localized in resting platelets, but became rapidly translocated to the proximity of the plasma membrane upon thrombin stimulation, thus indicating a role for PLD in platelet activation. PMID- 18432524 TI - Hydroxycarbamide associated platelet count oscillations in a patient with polycythaemia vera. A case report and review of the literature. AB - We describe an unusual case of oscillating platelet counts in a patient with polycythaemia vera. Following commencement of cytoreductive hydroxycarbamide therapy, episodes of thrombocytopenia were followed regularly by thrombocytosis. Platelet counts fluctuated periodically between approximately 200 and 800 x 109/l, with a 28 day cycle duration. Frequent adjustment of the hydroxycarbamide dose was not successful in preventing the oscillations in platelet count. In contrast, maintenance of a constant dose led to a gradual damping of the cycles and thus termination of the large oscillations. The case further implicates hydroxycarbamide as a potential cause of cyclic variations in platelet counts, and demonstrates that cessation of this drug is not always necessary in order to treat this phenomenon. PMID- 18432525 TI - The effect of gestational age on platelet surface expression of CD62P in preterm newborns. AB - Hemostasis in preterm newborns is characterized by low reserve functional capacity with special reference to the presence of such risk factors as asphyxia or infection. Platelets play vitally important role in hemostasis. Expression of CD62P is a marker of stimulated or activated blood platelets. The study involved a group of 16 preterm newborns, five girls and 11 boys. DAKO QIFIKIT was applied to calculate the number of these antigens. The mean CD 62P expression was found to be 23,792 per platelet. Correlation was found between antigen density and gestational age r = 0.954, p = 0.01. Evident deficit of P-selectin on the surface of platelets in preterm newborns may be at least in part responsible for platelet dysfunction, with special reference to interaction between circulating leukocytes and combating infection. PMID- 18432523 TI - Expression of complement components and inhibitors on platelet microparticles. AB - Platelet microparticles (PMP) are released from activated platelets and play an important role in hemostasis, thrombosis and inflammation. Since platelets were recently found to demonstrate an intrinsic capacity for activating both classical and alternative pathways of the complement system, the present study extended these observations to PMP. PMP were generated by treating platelets with 10 microM A23187 (37 degrees C, 5 min). PMP were identified by flow cytometry, based on size, Annexin V binding, and expression of P-selectin and GPIIb (CD41). PMP expressed gC1qR/p33, a multifunctional cellular protein that was recently described to activate the classical complement cascade. PMP also expressed the classical pathway and contact system regulator, C1 inhibitor (C1-INH), as well as CD55 and CD59. Despite C1-INH expression, PMP supported classical pathway C4 activation in the presence of purified C1 and C4. Moreover, statistically significant deposition of C3b and C5b-9 was detected on PMP exposed to plasma, concurrently with expression of CD55 and CD59. These data provide the first evidence for the ability of PMP to support in situ complement activation. Complement activation contributes to a variety of vascular and inflammatory disease states including atherosclerosis and ischemia/reperfusion injury. PMID- 18432526 TI - Controversies in chronic abacterial prostatitis/pelvic pain syndrome. AB - Chronic abacterial prostatitis (or prostatitis Category III according to the National Institutes of Health terminology) is subject to re-evaluation. Organizations such as the International Continence Society, the International Association for the Study of Pain and the European Association of Urology have been involved in a continuous endeavour to design a more accurate taxonomy system. The term prostate pain syndrome (PPS), as a subcategory of chronic pelvic pain syndromes, has recently been suggested as the most relevant. This condition is common, very bothersome and enigmatic. Symptoms are usually prolonged and, generally speaking, treatment results are unsatisfactory. During the last decade, research has focused on the distress caused by the condition, but although our knowledge has certainly increased there have been no real breakthroughs; controversies and many unanswered questions remain. Examples of pertinent issues are as follows: the criteria that need to be fulfilled for a diagnosis; the significance or lack of significance of leucocytes in expressed prostatic secretion; and the association between erectile dysfunction and PPS. The fundamental question remains: is the prostate really involved in the majority of men with PPS? This article focuses on some of the controversies. PMID- 18432527 TI - Nerve distribution in rat urinary bladder after incorporation of acellular matrix graft or subtotal cystectomy. AB - OBJECTIVE: In the treatment of reduced bladder capacity, matrix grafts have been used as a scaffold into which cell elements from the native bladder grow, eventually forming a new bladder segment. Functioning motor nerve endings in such segments in the rat have been demonstrated, although little is known about nerve distribution. We compare the pattern of nerve distribution in scaffold-augmented rat bladders with that in bladders regrown after subtotal cystectomy and that in control bladders. MATERIAL AND METHODS: Female Sprague-Dawley rats were either subtotally cystectomized (n=7) or had a part of the bladder dome replaced by an acellular collagen (small intestinal submucosa) matrix graft (n=10). Fourteen age matched, unoperated animals were used as controls. Two and a half to 10 months after surgery the bladders were stained for acetylcholinesterase and studied in wholemounts. RESULTS: No ganglion neurons were observed in any of the bladders. On their ventral side the control bladders showed longitudinal nerve trunks, running in parallel along the longitudinally oriented muscle bundles, while on the lateral and dorsal aspects the nerves were thinner, more irregularly arranged and frequently branched. In the bladders regrown after subtotal cystectomy, the ventral nerves were seen running obliquely to the still longitudinally oriented muscle bundles, resembling the pattern of the normal bladder base; the pattern of the dorsolateral nerves was the same as that in the controls. In the matrix bladders, the muscle and nerve patterns in the native part were the same as those in controls. Muscle bundles were growing into the matrix, accompanied by nerves, which showed limited branching when entering the matrix, usually running in parallel to the muscle, but then branching within the matrix. CONCLUSIONS: The nerves in the matrix grafts and the regrown parts of the subtotally cystectomized bladders derive from preexisting nerves in the bladder. In neither case does the nerve trunk or muscle bundle arrangement fully attain the pattern found in normal bladders. PMID- 18432529 TI - Body mass index and prognostic markers at radical prostatectomy. AB - OBJECTIVE: Obesity is increasing rapidly in Western countries. Approximately 40% of adult Danes are overweight and approximately 15% of these are obese. Epidemiological studies of obesity in relation to prostate cancer have provided conflicting results. Therefore, we examined correlations between body mass index (BMI) and clinicopathological prognostic markers, biochemical recurrence and operative morbidity in patients who had undergone radical prostatectomy. MATERIAL AND METHODS: The sample consisted of 293 Danish men treated with radical prostatectomy between 2000 and 2005 at Aarhus University Hospital. BMI was calculated as an indicator of obesity. Prospectively collected clinical and pathologic data from this population were used. RESULTS: The median BMI value was 26.2 kg/m(2) (range 19.6-41.7 kg/m(2)), which is slightly above the upper limit of normal. Currently accepted prognostic markers, such as prostate-specific antigen level, Gleason score and pT class, showed no statistically significant correlations with BMI. Patients with biochemical recurrence were evenly distributed among four different BMI quartiles and there was no difference in the length of hospitalization, indicating no differences in pre- or postoperative morbidity. Computations were repeated using only patients with the lowest (19.6 21.3 kg/m(2)) and highest (34.2-41.7 kg/m(2)) BMI values but statistically significant correlations were still not found. CONCLUSIONS: Several American studies have shown that obesity can lead to prostate cancer becoming more aggressive. The results of the present study involving a Danish prostate cancer population do not substantiate this or suggest any connection between BMI and operative morbidity. A possible explanation is that Danish obesity problems are not yet as severe as those in the USA. PMID- 18432528 TI - Parenteral estrogen versus combined androgen deprivation in the treatment of metastatic prostatic cancer: part 2. Final evaluation of the Scandinavian Prostatic Cancer Group (SPCG) Study No. 5. AB - OBJECTIVE: To compare parenteral estrogen therapy in the form of high-dose polyestradiol phosphate (PEP; Estradurin) with combined androgen deprivation (CAD) in the treatment of prostate cancer patients with skeletal metastases. The aim of the study was to compare anticancer efficacy and adverse events, especially cardiovascular events. MATERIAL AND METHODS: In total, 910 eligible patients with T0-4, NX, M1, G1-3 prostate cancer with an Eastern Cooperative Oncology Group performance status of 0-2 were randomized to treatment with either PEP 240 mg i.m. twice a month for 2 months and thereafter monthly, or flutamide (Eulexin) 250 mg t.i.d. per os in combination with either triptorelin (Decapeptyl) 3.75 mg i.m. per month or on an optional basis bilateral orchidectomy. RESULTS: At this final evaluation of the trial 855 of the 910 patients were dead. There was no difference between the treatment groups in terms of biochemical or clinical progression-free survival or in overall or disease specific survival. There was no difference in cardiovascular mortality, but a significant increase in non-fatal cardiovascular events in the PEP arm (p<0.05) predominantly caused by an increase in ischemic heart and heart decompensation events. There were 18 grave skeletal events in the CAD group but none in the PEP group (p=0.001). CONCLUSIONS: PEP has an anticancer efficacy equal to CAD and does not increase cardiovascular mortality in metastasized patients, but carries a significant risk of non-fatal cardiovascular events, which should be balanced against the skeletal complications in the CAD group. It is feasible to use Estradurin in the primary or secondary endocrine treatment of metastasized patients without prominent cardiac risk factors and especially those with osteoporosis. PMID- 18432530 TI - Histological classification and stage of newly diagnosed bladder cancer in a population-based study from the Northeastern United States. AB - OBJECTIVE: There are limited data on the distribution of bladder cancers in the general population, classified by World Health Organization (WHO)/International Society of Urological Pathology (ISUP) criteria. This study evaluated the classification and stage of bladder cancers as part of a population-based epidemiological study of bladder cancer in the Northeastern United States. MATERIAL AND METHODS: All New Hampshire residents with bladder cancer newly diagnosed from 1998 to 2000 were identified through the state cancer registry. All slides were reviewed by a single pathologist. Tumors were classified by two sets of standard criteria. RESULTS: The retrieval rate for cases was over 90%. Of 342 cases reviewed, 15 were excluded for technical reasons or because malignancy was not definitively diagnosed. According to WHO/ISUP criteria, 25.7% of tumors were papillary urothelial neoplasms of low malignant potential (PUNLMP), 34.3% low-grade papillary carcinomas, 22.6% high-grade papillary carcinomas, 10.1% non papillary urothelial carcinomas and 5.5% carcinoma in situ. By WHO (1973) criteria, 52.5% of tumors were grade 1, 21.4% grade 2 and 26.1% grade 3. Two thirds of all tumors were stage Ta, 20.8% stage T1 and 7.6% stage >or=T2. 100% of PUNLMPs were non-invasive, 6.3% of low-grade carcinomas were invasive and 64.9% of high-grade carcinomas were invasive. CONCLUSIONS: Compared to clinic or hospital referral-based series, this study documents a higher percentage of non invasive tumors and a lower percentage of muscle-invasive tumors. There was also a higher percentage of PUNLMP tumors and fewer high-grade papillary carcinomas than in other series. These results may more accurately reflect prevalence data for bladder cancer grade and stage, although geographic variability may exist. PMID- 18432531 TI - Outcome of very large superficial bladder tumours: a 10-year experience. AB - OBJECTIVES: To determine the biological behaviour of very large superficial bladder tumours (pTa, pT1) and evaluate the impact of the initial tumour weight on long-term prognosis. MATERIAL AND METHODS: Of 1569 patients who presented with bladder tumours over a 10-year period, 1070 of the tumours were superficial. Fifty-nine patients had very large tumours (resected weight >or= 15 g). Case notes were analysed to determine recurrence, progression and survival. Median follow-up was 60 months (range 1-156 months). Histological slides were reviewed for all tumours initially reported as pT1 to determine the presence of uninvolved muscle. Statistical analysis was performed using the Kaplan-Meier method to calculate progression and survival estimates. RESULTS: The overall progression and recurrence rates for very large superficial bladder tumours were 18% and 68%, respectively. The progression rates for Ta, T1, G1, G2 and G3 tumours were 4%, 28%, 0%, 20% and 50%, respectively, with highest progression rates being seen for pT1G2 and pT1G3 tumours. The progression rate was significantly influenced by initial stage (p=0.01) and grade (p=0.03). Tumour weight did not affect either recurrence, progression or cause-specific survival. There were no differences in progression and survival rates in patients with tumour weights of 15-30 and >30 g (p=0.80 and 0.07, respectively). The review of histology slides of T1 tumours showed that 7/10 cases (70%) with progression had no muscle or an inadequate amount of muscle for definitive staging. Upper urinary tract tumours were seen in only two patients (3.4%). CONCLUSIONS: Large size is not an adverse prognostic factor for patients with a superficial bladder tumour. However, these cases are difficult to stage. In view of the high rates of progression and disease-specific mortality, we recommend that very large pT1G2 bladder tumours should be considered as high-risk tumours and targeted for aggressive treatment, including early re-resection, to rule out any occult invasive disease. PMID- 18432532 TI - Relation between radionuclide imaging and pathologic findings of ureteropelvic junction obstruction in neonatal hydronephrosis. AB - OBJECTIVE: An excessive amount of collagen fibers around the muscle cells in the ureteropelvic junction could be responsible for obstruction in patients with hydronephrosis. We aimed to elucidate the ultrastructure of the ureters and correlate this finding with the prognostic outcome and to correlate the histopathological findings with diuretic radionuclide renography findings. MATERIAL AND METHODS: Biopsy specimens of 20 children who underwent dismembered pyeloplasty for ureteropelvic junction obstruction were analyzed. The patients were grouped according to their age: infants (<12 months) and others (>12 months). Diuretic radionuclide imaging was performed using (99m)Tc mercaptylacetyltriglycine in the pre- and postoperative periods. Changes in differential renal function and excretion patterns on diuretic renography were evaluated in relation to the findings noted on histopathological examination of the biopsy specimens. Excretion patterns were classified as follows: A, normal; B, responsive to diuretic; C, minimal response to diuretic with some excretion after postural change; and D, very poor/no drainage despite diuretics. Biopsy materials were analyzed for the presence and extent of inflammation, fibrosis and changes in the smooth muscle layer using Masson's trichrome stain and immunohistochemical staining. Histopathological findings were graded from zero to three, depending on severity. RESULTS: In patients aged <12 months, preoperative differential renal function (DRF) was associated with fibrosis (F) and smooth muscle hypertrophy (SMH) [mean (SD) DRF for both F and SMH were Grade 0-1, 47.8% (6.4%); Grade 2-3, 36.2% (11.3%); p<0.05]; and change in DRF was associated with inflammation [Grade 0-1, -0.1% (4.0%); Grade 2-3, 5.8% (3.0%); p<0.05]. Excretion patterns or improvement in excretion were not associated with any of the histopathological features. Change in DRF was significantly associated with inflammation Grade 2-3 (beta coefficient, 5.8; 95% CI 1.4-10.3). CONCLUSIONS: Histopathological evaluation of renal parenchymal biopsy specimens obtained during pyeloplasty may be useful to provide an objective method for predicting the recovery of renal function. In addition, this will allow comparison of the types of histopathological alterations with the changes in differential renal function in order to predict the potential final improvement. PMID- 18432533 TI - The prevalence of infrequent bedwetting and nocturnal enuresis in childhood. A large British cohort. AB - OBJECTIVE: Although epidemiological surveys generally indicate declining rates of bedwetting with age, recent studies show that children with severe nocturnal enuresis have lower prevalence rates, which persist into adolescence. This study reports prevalence rates of both infrequent bedwetting (IB) and nocturnal enuresis (NE) at five time points during childhood with a large cohort of children. MATERIAL AND METHODS: The Avon Longitudinal Study of Parents and Children provided a cohort of 13,973 singleton/twin infants alive at 12 months. These were followed up at 54, 65, 78, 91 and 115 months with a questionnaire relating to frequency of bedwetting and other variables. The prevalences of IB and NE were derived from these measures with missing data imputation being used to correct for possible loss-to-follow-up bias. RESULTS: The overall prevalence rate of bedwetting declined from 30% (54 months) to 9.5% (115 months), being most pronounced between 54 and 65 months. Children with NE (wet at least twice a week) had lower prevalence rates at all ages but were more likely to persist with the problem over time and to have the non-monosymptomatic type of bedwetting compared to children with IB. CONCLUSIONS: Children with the severest form of bedwetting are likely to persist with the problem and to have the more complex form (non monosymptomatic). The results are discussed in relation to the clinical importance of early identification. PMID- 18432534 TI - Normal voiding patterns assessed by means of a frequency-volume chart. AB - OBJECTIVE: Voiding dysfunction is one of the commonest problems among the elderly. This study aimed to elucidate the mechanisms behind the diurnal and nocturnal urine output patterns in young and elderly male volunteers. Of particular interest was bladder reservoir function during the day and night. MATERIAL AND METHODS: Young males (n=25; median age 25.0 years; range 22-32 years) and elderly males (n=18; median age 61.1 years; range 55-73 years) were included in the study. Their voiding habits were assessed from a 3-day frequency volume chart (FVC) detailing all fluid intake and urine output. Data on voided volume, voiding frequency, maximum voided volume (MVV) and average voided volume (AVV) were obtained from the FVC. RESULTS: The young males' average fluid intake was significantly higher than that of the elderly males. On average the elderly males had a slightly higher voiding frequency than the young males, although this was not statistically significant. There was no difference between the two groups regarding their AVVs at night (including the first morning void). The MVVs of the young and elderly males were significantly different, whereas the ratio between voided volume and MVV did not differ between the groups. CONCLUSIONS: The main finding was that young and elderly males void with equal average volumes of urine at night (first morning void inclusive), whereas the elderly void with smaller volumes than the young during the day-time. The elderly thus seem to have the capacity to adjust their AVV to night-time urine output size. PMID- 18432535 TI - Three-years experience with Neoral C2 monitoring adjusted to a target range of 500-600 ng/ml in long-term renal transplant recipients receiving dual immunosuppressive therapy. AB - OBJECTIVE: There is little knowledge about the optimal 2-h post-dose concentration (C(2) level) of cyclosporin A (CsA) in renal transplant recipients beyond 1 year post-transplant. The aim of this study was to investigate the effects of C(2)-CsA monitoring on Neoral dose, renal graft function and systemic blood pressure in long-term renal transplant recipients, who had previously been monitored by means of trough levels (C(0)). MATERIAL AND METHODS: Eighty-six patients treated with CsA+prednisolone were reviewed retrospectively during a follow-up period after switching to C(2)-CsA monitoring. RESULTS: The patients were 6.0 years (3.4, 9.0 years) [median (25% quartile, 75% quartile)] post transplant at the time of conversion to C(2)-CsA monitoring. They were studied for 3.7 years (3.3, 3.8 years). Baseline C(0)-CsA level was 161 ng/ml (131, 208 ng/ml). The Neoral dose was reduced in 95% of the recipients. The median C(2) level was reduced by 40% to 585 ng/ml (484, 670 ng/ml) and, accordingly, the Neoral dose was reduced by 30% to 2.8 mg/kg/day (2.3, 3.8 mg/kg/day). Overall, plasma creatinine remained stable during the follow-up period. In 48/86 patients (56%), the plasma creatinine level was lower at the end of the study compared to baseline, declining from 163 micromol/l (124, 189 micromol/l) in 2001 to 147 micromol/l (106, 172) in 2005. Three patients (3.5%) had late acute rejections, 14 (16.3%) discontinued CsA, five (5.8%) commenced dialysis and seven (8.1%) died. CONCLUSION: Adoption of C(2)-CsA monitoring resulted in a substantial reduction in Neoral dose, while the overall renal graft function remained stable. PMID- 18432536 TI - Rising from a chair: a simple screening test for physical function in predialysis patients. AB - OBJECTIVES: Patients with chronic end-stage renal diseases have decreased physical fitness. This physical deconditioning is relative but the consequences in daily life are related to functional thresholds. The main purpose of this study was to gather information about physical performance, physical fitness and activity in predialysis patients. A second aim was to investigate the possibility of identifying patients at risk of physical deconditioning using clinical tests. MATERIAL AND METHODS: Fifty-five predialysis patients with a glomerular filtration rate (GFR) of